Bacterial Hand Contamination and Transfer after Use of Contaminated Bulk-Soap-Refillable Dispensers▿†

PubMed Central

Zapka, Carrie A.; Campbell, Esther J.; Maxwell, Sheri L.; Gerba, Charles P.; Dolan, Michael J.; Arbogast, James W.; Macinga, David R.

2011-01-01

Bulk-soap-refillable dispensers are prone to extrinsic bacterial contamination, and recent studies demonstrated that approximately one in four dispensers in public restrooms are contaminated. The purpose of this study was to quantify bacterial hand contamination and transfer after use of contaminated soap under controlled laboratory and in-use conditions in a community setting. Under laboratory conditions using liquid soap experimentally contaminated with 7.51 log10 CFU/ml of Serratia marcescens, an average of 5.28 log10 CFU remained on each hand after washing, and 2.23 log10 CFU was transferred to an agar surface. In an elementary-school-based field study, Gram-negative bacteria on the hands of students and staff increased by 1.42 log10 CFU per hand (26-fold) after washing with soap from contaminated bulk-soap-refillable dispensers. In contrast, washing with soap from dispensers with sealed refills significantly reduced bacteria on hands by 0.30 log10 CFU per hand (2-fold). Additionally, the mean number of Gram-negative bacteria transferred to surfaces after washing with soap from dispensers with sealed-soap refills (0.06 log10 CFU) was significantly lower than the mean number after washing with contaminated bulk-soap-refillable dispensers (0.74 log10 CFU; P < 0.01). Finally, significantly higher levels of Gram-negative bacteria were recovered from students (2.82 log10 CFU per hand) than were recovered from staff (2.22 log10 CFU per hand) after washing with contaminated bulk soap (P < 0.01). These results demonstrate that washing with contaminated soap from bulk-soap-refillable dispensers can increase the number of opportunistic pathogens on the hands and may play a role in the transmission of bacteria in public settings. PMID:21421792

Bacterial hand contamination and transfer after use of contaminated bulk-soap-refillable dispensers.

PubMed

Zapka, Carrie A; Campbell, Esther J; Maxwell, Sheri L; Gerba, Charles P; Dolan, Michael J; Arbogast, James W; Macinga, David R

2011-05-01

Bulk-soap-refillable dispensers are prone to extrinsic bacterial contamination, and recent studies demonstrated that approximately one in four dispensers in public restrooms are contaminated. The purpose of this study was to quantify bacterial hand contamination and transfer after use of contaminated soap under controlled laboratory and in-use conditions in a community setting. Under laboratory conditions using liquid soap experimentally contaminated with 7.51 log(10) CFU/ml of Serratia marcescens, an average of 5.28 log(10) CFU remained on each hand after washing, and 2.23 log(10) CFU was transferred to an agar surface. In an elementary-school-based field study, Gram-negative bacteria on the hands of students and staff increased by 1.42 log(10) CFU per hand (26-fold) after washing with soap from contaminated bulk-soap-refillable dispensers. In contrast, washing with soap from dispensers with sealed refills significantly reduced bacteria on hands by 0.30 log(10) CFU per hand (2-fold). Additionally, the mean number of Gram-negative bacteria transferred to surfaces after washing with soap from dispensers with sealed-soap refills (0.06 log(10) CFU) was significantly lower than the mean number after washing with contaminated bulk-soap-refillable dispensers (0.74 log(10) CFU; P < 0.01). Finally, significantly higher levels of Gram-negative bacteria were recovered from students (2.82 log(10) CFU per hand) than were recovered from staff (2.22 log(10) CFU per hand) after washing with contaminated bulk soap (P < 0.01). These results demonstrate that washing with contaminated soap from bulk-soap-refillable dispensers can increase the number of opportunistic pathogens on the hands and may play a role in the transmission of bacteria in public settings.

Lytic bacteriophages reduce Escherichia coli O157

PubMed Central

Ferguson, Sean; Roberts, Cheryl; Handy, Eric; Sharma, Manan

2013-01-01

The role of lytic bacteriophages in preventing cross contamination of produce has not been evaluated. A cocktail of three lytic phages specific for E. coli O157:H7 (EcoShield™) or a control (phosphate buffered saline, PBS) was applied to lettuce by either; (1) immersion of lettuce in 500 ml of EcoShield™ 8.3 log PFU/ml or 9.8 log PFU/ml for up to 2 min before inoculation with E. coli O157:H7; (2) spray-application of EcoShield™ (9.3 log PFU/ml) to lettuce after inoculation with E. coli O157:H7 (4.10 CFU/cm2) following exposure to 50 μg/ml chlorine for 30 sec. After immersion studies, lettuce was spot-inoculated with E. coli O157:H7 (2.38 CFU/cm2). Phage-treated, inoculated lettuce pieces were stored at 4°C for and analyzed for E. coli O157:H7 populations for up to 7 d. Immersion of lettuce in 9.8 log PFU/ml EcoShield™ for 2 min significantly (p < 0.05) reduced E. coli O157:H7 populations after 24 h when stored at 4°C compared with controls. Immersion of lettuce in suspensions containing high concentrations of EcoShield™ (9.8 log PFU/ml) resulted in the deposition of high concentrations (7.8 log log PFU/cm2) of bacteriophages on the surface of fresh cut lettuce, potentially contributing to the efficacy of the lytic phages on lettuce. Spraying phages on to inoculated fresh cut lettuce after being washed in hypochlorite solution was significantly more effective in reducing E. coli O157:H7 populations (2.22 log CFU/cm2) on day 0 compared with control treatments (4.10 log CFU/cm2). Both immersion and spray treatments provided protection from E. coli O157:H7 contamination on lettuce, but spray application of lytic bacteriophages to lettuce was more effective in immediately reducing E. coli O157:H7 populations fresh cut lettuce. PMID:23819106

Bacteriological quality of drinking water in the Atebubu-Amantin District of the Brong-Ahafo Region of Ghana

NASA Astrophysics Data System (ADS)

Tekpor, M.; Akrong, M. O.; Asmah, M. H.; Banu, R. A.; Ansa, E. D. O.

2017-09-01

The study was carried out to determine the bacteriological safety of water in hand-dug wells in the Atebubu-Amantin District of the Brong-Ahafo Region in Ghana. A total of 60 samples were collected from ten hand dug wells and analysed for total coliform (TC), faecal coliform (FC), E. coli (EC), Salmonella spp. (SP) and Enterococcus spp. (ES). Data was collected in both the rainy and the dry seasons. The results obtained showed that water from all the wells in the study area did not meet the World Health Organisation guideline and Ghana standard for drinking water of zero (0) coliform forming unit (cfu) per 100 ml for TC, FC, EC, SP and ES, respectively. Contamination was found to be high in the wells during the wet season as compared to the dry season. Wells (A1 to A5) which were close to septic tanks had high bacteria counts in both seasons. The total coliform counts ranged from 2.98 to 5.93 log cfu/100 ml in the wet season and 3.10-5.03 log cfu/100 ml in the dry season. There was drastic reduction of faecal coliform count from a range of 2.78-4.55 log cfu/100 ml in the wet season to 1.70-3.51 log cfu/100 ml in the dry season. The high bacteria count in wells A1 to A5 could be attributed to the closeness of the wells to the septic tank, and contaminant transport through the saturated underground zones. It is recommended that the water should be treated properly before drinking.

Survival and growth of Yersinia enterocolitica strains inoculated in skimmed milk treated with high hydrostatic pressure.

PubMed

De Lamo-Castellví, Sílvia; Roig-Sagués, Artur X; Capellas, Marta; Hernández-Herrero, Manuela; Guamis, Buenaventura

2005-07-25

Four human pathogenic strains of Yersinia enterocolitica (serotypes O:1, O:3, O:8, and O:9) were inoculated (7-8 log CFU/ml) in UHT skimmed milk and treated at 300, 400, and 500 MPa for 10 min at 20 degrees C, and then kept at 8 degrees C to assess their evolution for 15 days. Treatments at 400 and 500 MPa caused the highest lethality, generally reaching counts below detection level (1 CFU/ml) in the culture media. At 300 MPa, the most baroresistant serotypes were O:3 and O:8. After 15 days of storage at 8 degrees C, Y. enterocolitica showed growth over 8 log (CFU/ml) in all treatments. Kinetic study of microbial inactivation in skimmed milk was performed with serotype O:8 at 300 MPa, showing a tailing after 35 min of pressure treatment.

Inhibition of Listeria monocytogenes by piscicolin 126 in milk and Camembert cheese manufactured with a thermophilic starter.

PubMed

Wan, J; Harmark, K; Davidson, B E; Hillier, A J; Gordon, J B; Wilcock, A; Hickey, M W; Coventry, M J

1997-03-01

The effect of bacteriocin, piscicolin 126, on the growth of Listeria monocytogenes and cheese starter bacteria was investigated in milk and in Camembert cheese manufactured from milk challenged with 10(2) cfu ml(-1) L. monocytogenes. In milk incubated at 30 degrees C, piscicolin 126 added in the range of 512-2,048 AU ml(-1) effectively inhibited growth of L. monocytogenes for more than 20 d when challenged with approximately 10(2) cfu ml(-1) L. monocytogenes. At higher challenge levels (10(4) and 10(6) cfu ml(-1)), piscicolin 126 reduced the viable count of L. monocytogenes by 4-5 log units immediately after addition of the bacteriocin; however, growth of Listeria occurred within 24 h. The minimum inhibitory concentration (MIC) of piscicolin 126 against lactic acid cheese starter bacteria was generally greater than 204,800 AU ml(-1) , and the viable count and acid production of these starter cultures in milk were not affected by the addition of 2,048 AU ml(-1) piscicolin 126. Camembert cheeses made from milk challenged with L. monocytogenes and with added piscicolin 126 showed a viable count of L. monocytogenes 3-4 log units lower than those without piscicolin 126. Inactivation of piscicolin 126 by proteolytic enzymes from cheese starter bacteria and mould together with the emergence of piscicolin 126-resistant isolates was responsible for the recovery of L. monocytogenes in the cheeses during ripening.

Assessment of the efficacy of the first water system for emergency hospital use.

PubMed

Long, Sharon C; Olstadt, Jeremy

2011-03-01

The First Water Responder B package water treatment device was evaluated for its ability to reduce the levels of spiked indicators and pathogens (Escherichia coli, MS2 coliphage, murine adenovirus, and Cryptosporidium oocysts) in a surface water to partially evaluate its appropriateness to be used to provide safe drinking water to hospitals during emergency situations. Lake water was collected in 50-L carboys and spiked with selected indicators and pathogens (E coli, MS2 coliphage, murine adenovirus, and Cryptosporidium oocysts) at 2 different spike levels (low and high). This water was treated using the First Water Responder B, and the microorganisms were enumerated before and after treatment using US Environmental Protection Agency and Standard Methods. Microbial removal efficiencies were compared with Environmental Protection Agency guidelines. E coli spikes ranged from 2.9 to 1059 colony-forming units (CFU)/100 mL with removals to below detection limits (1 CFU/100 mL) to 2.8 CFU/100 mL or 0.98 to 3.5 log(10) reductions. MS2 coliphage spikes ranged from 3 plaque-forming units (PFU) to 837 PFU/100 mL with removals to below detection limits (1 PFU/100 mL) to 11.7 PFU/100 mL or 0.65 to 1.9 log(10) reductions. Murine adenovirus spikes ranged from 203 to 8410 most probable number (MPN) of infectious units/100 mL with removals to below detection limits (23 MPN infectious units/100 mL) to 1370 MPN infectious units/100 mL or 0.79 to >1.2 log(10) reductions. Cryptosporidium parvum oocyst spikes ranged from 52 to 853 oocysts per liter with removals to below detection limits (<1 oocyst per liter) to 0.3 oocysts per liter or >2.2 to 3.4 log(10) reductions. Although the First Water system could remove a significant portion of the spiked organisms, it is recommended that this point-of-use system be coupled with chemical disinfection in a multiple-barrier approach to provide water of the highest reasonably achievable quality for hospital use in emergency situations. ©2011 American Medical Association. All rights reserved.

Inactivation of Escherichia coli O157:H7 on Orange Fruit Surfaces and in Juice Using Photocatalysis and High Hydrostatic Pressure.

PubMed

Yoo, Sungyul; Ghafoor, Kashif; Kim, Jeong Un; Kim, Sanghun; Jung, Bora; Lee, Dong-Un; Park, Jiyong

2015-06-01

Nonpasteurized orange juice is manufactured by squeezing juice from fruit without peel removal. Fruit surfaces may carry pathogenic microorganisms that can contaminate squeezed juice. Titanium dioxide-UVC photocatalysis (TUVP), a nonthermal technique capable of microbial inactivation via generation of hydroxyl radicals, was used to decontaminate orange surfaces. Levels of spot-inoculated Escherichia coli O157:H7 (initial level of 7.0 log CFU/cm(2)) on oranges (12 cm(2)) were reduced by 4.3 log CFU/ml when treated with TUVP (17.2 mW/cm(2)). Reductions of 1.5, 3.9, and 3.6 log CFU/ml were achieved using tap water, chlorine (200 ppm), and UVC alone (23.7 mW/cm(2)), respectively. E. coli O157:H7 in juice from TUVP (17.2 mW/cm(2))-treated oranges was reduced by 1.7 log CFU/ml. After orange juice was treated with high hydrostatic pressure (HHP) at 400 MPa for 1 min without any prior fruit surface disinfection, the level of E. coli O157:H7 was reduced by 2.4 log CFU/ml. However, the E. coli O157:H7 level in juice was reduced by 4.7 log CFU/ml (to lower than the detection limit) when TUVP treatment of oranges was followed by HHP treatment of juice, indicating a synergistic inactivation effect. The inactivation kinetics of E. coli O157:H7 on orange surfaces followed a biphasic model. HHP treatment did not affect the pH, °Brix, or color of juice. However, the ascorbic acid concentration and pectinmethylesterase activity were reduced by 35.1 and 34.7%, respectively.

Fermentation of wheat: effects of backslopping different proportions of pre-fermented wheat on the microbial and chemical composition.

PubMed

Moran, Colm A; Scholten, Ronald H J; Tricarico, Juan M; Brooks, Peter H; Verstegen, Martin W A

2006-04-01

The objective of the study was to examine effect of backslop on the chemical and microbiological characteristics of fermented wheat (FW). Coarsely ground wheat was mixed with water (1:3 wt/wt) and inoculated with 6 log cfu ml(-1) each of an overnight culture of Lactobacillus plantarum and Pediococcus pentosaceus. Four fermentation treatments were conducted in 45 1, closed, PVC containers over 48 hours. Three treatments investigated the benefits of the addition of previously fermented wheat (backslopping, BSL) at different proportions (0.20, 0.33 or 0.42 kg) to freshly prepared wheat. The control treatment contained no addition of BSL. Elimination of coliforms from the FW within 48 h was only achieved through backslopping; where coliform bacteria counts decreased from approximately 6.5 log10 cfu ml(-1) to less than 3 log10 cfu ml(-1). There was no apparent advantage in increasing the backslop proportion above 0.20. However, the exclusion of coliform bacteria required the pH to remain below 4.0 for at a minimum of 24 h. The results of these studies indicate that fermentation of wheat has the potential to reduce the risk of feed-borne colibacillosis and provides a practical alternative to producers that cannot ferment multiple diets or have limited fermentation capacity.

Development and validation of an in vitro pharmacokinetic/pharmacodynamic model to test the antibacterial efficacy of antibiotic polymer conjugates.

PubMed

Azzopardi, Ernest A; Ferguson, Elaine L; Thomas, David W

2015-04-01

This study describes the use of a novel, two-compartment, static dialysis bag model to study the release, diffusion, and antibacterial activity of a novel, bioresponsive dextrin-colistin polymer conjugate against multidrug resistant (MDR) wild-type Acinetobacter baumannii. In this model, colistin sulfate, at its MIC, produced a rapid and extensive drop in viable bacterial counts (<2 log10 CFU/ml at 4 h); however, a marked recovery was observed thereafter, with regrowth equivalent to that of control by 48 h. In contrast, dextrin-colistin conjugate, at its MIC, suppressed bacterial growth for up to 48 h, with 3 log10 CFU/ml lower bacterial counts after 48 h than those of controls. Doubling the concentration of dextrin-colistin conjugate (to 2× MIC) led to an initial bacterial killing of 3 log10 CFU/ml at 8 h, with a similar regrowth profile to 1× MIC treatment thereafter. The addition of colistin sulfate (1× MIC) to dextrin-colistin conjugate (1× MIC) resulted in undetectable bacterial counts after 4 h, followed by suppressed bacterial growth (3.5 log10 CFU/ml lower than that of control at 48 h). Incubation of dextrin-colistin conjugates with infected wound exudate from a series of burn patients (n = 6) revealed an increasing concentration of unmasked colistin in the outer compartment (OC) over time (up to 86.3% of the initial dose at 48 h), confirming that colistin would be liberated from the conjugate by endogenous α-amylase within the wound environment. These studies confirm the utility of this model system to simulate the pharmacokinetics of colistin formation in humans administered dextrin-colistin conjugates and further supports the development of antibiotic polymer conjugates in the treatment of MDR infections. Copyright © 2015, American Society for Microbiology. All Rights Reserved.

Isolation, characterization and screening of rhizospheric bacteria of Pittosferum resiniferum Hemsl.

NASA Astrophysics Data System (ADS)

Hanirah, R.; Piakong, M. T.; Syaufi, L.

2015-04-01

The bacterial rhizosphere species of host plant, Petroleum Nut (Pittosferum resiniferum) were isolated and characterized morphologically. The isolates were designated as, TSArp- Cr2, TSArp- Cr3, TSArp- Cr4, TSArp- Cr5, TSArp- Cr6 and TSArp- Cr7. All of the species were tested on three different concentration of phenol (1mM, 3mM and 5mM). Only species TSArp- Cr4 and TSArp- Cr6 growth were detected. The highest growth is 6Log10CFU/ml in 1mM by TSA-Cr4. The lowest reading was 3.6 Log10CFU/ml in 3mM by TSA-Cr6. Species TSArp- Cr4 has higher tolerance on phenol compared to TSArp- Cr6

Effects of Plant-Derived Extracts, Other Antimicrobials, and Their Combinations against Escherichia coli O157:H7 in Beef Systems.

PubMed

Ko, Kyung Yuk; Geornaras, Ifigenia; Paik, Hyun-Dong; Kim, Kee-Tae; Sofos, John N

2015-06-01

The antimicrobial effects of thyme oil (TO), grapefruit seed extract (GSE), and basil essential oil, alone or in combination with cetylpyridinium chloride (CPC), sodium diacetate, or lactic acid, were evaluated against Escherichia coli O157:H7 in a moisture-enhanced beef model system. The model system was composed of a nonsterile beef homogenate to which NaCl (0.5%) and sodium tripolyphosphate (0.25%) were added, together with the tested antimicrobial ingredients. Beef homogenate treatments were inoculated (ca. 3 log CFU/ml) with rifampin-resistant E. coli O157:H7 (eight-strain mixture) and incubated at 15 °C (48 h). The most effective individual treatments were TO (0.25 or 0.5%) and GSE (0.5 or 1.0%), which immediately reduced (P < 0.05) pathogen levels by ≥ 3.4 log CFU/ml. Additionally, CPC (0.04%) reduced initial E. coli O157:H7 counts by 2.7 log CFU/ml. Most combinations of the tested plant-derived extracts with CPC (0.02 or 0.04%) and sodium diacetate (0.25%) had an additive effect with respect to antibacterial activity. In a second study, antimicrobial interventions were evaluated for their efficacy in reducing surface contamination of E. coli O157:H7 on beef cuts and to determine the effect of these surface treatments on subsequent internalization of the pathogen during blade tenderization. Beef cuts (10 by 8 by 3.5 cm) were inoculated (ca. 4 log CFU/g) on one side with the rifampin-resistant E. coli O157:H7 strain mixture and were then spray treated (20 lb/in(2), 10 s) with water, GSE (5 and 10%), lactic acid (5%), or CPC (5%). Untreated (control) and spray-treated surfaces were then subjected to double-pass blade tenderization. Surface contamination (4.4 log CFU/g) of E. coli O157:H7 was reduced (P < 0.05) to 3.4 (5% CPC) to 4.1 (water or 5% GSE) log CFU/g following spray treatment. The highest and lowest transfer rates of pathogen cells from the surface to deeper tissues of blade-tenderized sections were obtained in the untreated control and CPC-treated samples, respectively.

Survival of Listeria monocytogenes in a simulated recirculating brine chiller system.

PubMed

Gailey, J K; Dickson, J S; Dorsa, W

2003-10-01

Contamination by Listeria monocytogenes of processed meats after cooking presents a significant food safety risk. The purpose of this study was to determine the survival of L. monocytogenes in a simulated recirculating brine chiller system using pH values of 5, 6, and 7 with free chlorine concentrations of 0, 3, 5, and 10 ppm in 20% salt brine at -12 degrees C. At pH values of 5, 6, and 7 with chlorine concentrations of 2 and 3 ppm, using 10(8) CFU in a test tube system, an immediate drop of 0.28 log CFU/ml with no significance between treatments (P > 0.05), followed by a steady survival phase with a slope close to 0, was observed. In brine at a pH of 5 with 5 and 10 ppm of chlorine, an initial drop of 0.8 log CFU/ml was observed, which was followed by a steady survival phase with a destruction slope close to zero. At an inoculation concentration of 10(2) CFU in a test tube system (pH values of 5 and 7 with 0 and 10 ppm of chlorine), the average initial drop for all treatments was 0.1 log CFU/ml, which was followed by a steady survival phase. In a recirculating system, very few cells were destroyed during the brine chilling process, but only low numbers of L. monocytogenes were recovered from the brine and uninoculated hot dogs. Although little destruction of L. monocytogenes was noted, the dilution effect observed during the study indicates that environmental contamination of a brine chiller system poses little danger of postcooking contamination for processed meats if the system is regularly cleaned and sanitized.

Efficacy of detergents in removing Salmonella and Shigella spp. from the surface of fresh produce.

PubMed

Raiden, Renee M; Sumner, Susan S; Eifert, Joseph D; Pierson, Merle D

2003-12-01

Fresh produce has been implicated in several foodborne disease outbreaks. Produce surfaces can be primary sites of contamination during production and handling. One approach to reduce contamination is to treat fresh produce with rinsing agents. In this study, different detergent agents were used at 22 and 40 degrees C to determine their efficacy in removing Salmonella and Shigella spp. from the surfaces of strawberries, tomatoes, and green-leaf lettuce. Produce was inoculated at 22 degrees C with a cocktail of nalidixic acid-resistant organisms (6 to 6.5 log CFU/ml). After air drying for 1 h, samples were rinsed with either 0.1% Tween 80, 0.1% sodium lauryl sulfate (SLS), or water (control) at 22 or 40 degrees C. Rinse solutions were spiral plated onto tryptic soy agar supplemented with 50 mg of nalidixic acid per liter. In trials involving strawberries and lettuce, Salmonella and Shigella were removed at levels of 4 and 3 log CFU/ml, respectively, except from Salmonella-inoculated strawberries rinsed with SLS, for which minimal removal rates were 1.5 log CFU/ml at 22 degrees C and < 1 log CFU/ml at 40 degrees C. When whole strawberries were analyzed after rinsing with SLS, few organisms were recovered. This result suggests that SLS may have a lethal or sublethal effect on Salmonella, especially when a 40 degrees C solution is used. Salmonella and Shigella removal rates for tomatoes were 1 and 1.5 log CFU/ml lower, respectively, than those for strawberries or lettuce. Overall, detergents were no more effective in removing organisms from produce than water was. The detergents examined would not constitute effective overall produce rinse treatments.

Viability of Listeria monocytogenes on uncured turkey breast commercially prepared with and without buffered vinegar during extended storage at 4 and 10°C.

PubMed

Porto-Fett, Anna C S; Campano, Stephen G; Shoyer, Bradley A; Wadsworth, Sarah; Luchansky, John B

2014-06-01

We determined the viability of Listeria monocytogenes on uncured turkey breast containing buffered vinegar (BV) and surface treated with a stabilized solution of sodium chlorite in vinegar (VSC). Commercially produced, uncured, deli-style turkey breast was formulated with BV (0.0, 2.0, 2.5, or 3.0%), sliced (ca. 100 g and ca. 1.25 cm thick), and subsequently surface inoculated (ca. 4.3 log CFU per slice) in each of two trials with a five-strain cocktail of L. monocytogenes. Next, 1 ml per side of a 2 or 10% solution of VSC was added to each package before vacuum sealing and storing at 4 or 10°C. Without antimicrobials, L. monocytogenes numbers increased by ca. 6.2 log CFU per slice after 90 and 48 days of storage at 4 or 10°C, respectively. At 4°C, L. monocytogenes numbers increased by ca. 0.4 to 1.9 log CFU per slice on turkey breast formulated with 2.0 or 2.5% BV and treated or not with 2% VSC, whereas when treated with 10% VSC, L. monocytogenes levels remained relatively unchanged over 90 days. However, when turkey breast was formulated with 3.0% BV and treated or not with VSC, pathogen numbers decreased by ca. 0.7 to 1.3 log CFU per slice. At 10°C, L. monocytogenes numbers increased by ca. 1.5 to 5.6 log CFU per slice after 48 days when formulated with 2.0 to 3.0% BV and treated or not with 2% VSC. When formulated with 2.0% BV and treated with 10% VSC, L. monocytogenes numbers increased by ca. 3.3 log CFU per slice, whereas when formulated with 2.5 or 3.0% BV and treated with 10% VSC, L. monocytogenes decreased by ca. 0.3 log CFU per slice. Inclusion of BV as an ingredient in uncured turkey breast, alone or in combination with VSC added to the package, appreciably suppressed outgrowth of L. monocytogenes during an extended refrigerated shelf life.

Antimicrobial effect of electrolyzed water for inactivating Campylobacter jejuni during poultry washing.

PubMed

Park, Hoon; Hung, Yen-Con; Brackett, Robert E

2002-01-30

The effectiveness of electrolyzed (EO) water for killing Campylobacter jejuni on poultry was evaluated. Complete inactivation of C. jejuni in pure culture occurred within 10 s after exposure to EO or chlorinated water, both of which contained 50 mg/l of residual chlorine. A strong bactericidal activity was also observed on the diluted EO water (containing 25 mg/l of residual chlorine) and the mean population of C. jejuni was reduced to less than 10 CFU/ml (detected only by enrichment for 48 h) after 10-s treatment. The diluted chlorine water (25 mg/l residual chlorine) was less effective than the diluted EO water for inactivation of C. jejuni. EO water was further evaluated for its effectiveness in reducing C. jejuni on chicken during washing. EO water treatment was equally effective as chlorinated water and both achieved reduction of C. jejuni by about 3 log10 CFU/g on chicken, whereas deionized water (control) treatment resulted in only 1 log10 CFU/g reduction. No viable cells of C. jejuni were recovered in EO and chlorinated water after washing treatment, whereas high populations of C. jejuni (4 log10 CFU/ml) were recovered in the wash solution after the control treatment. Our study demonstrated that EO water was very effective not only in reducing the populations of C. jejuni on chicken, but also could prevent cross-contamination of processing environments.

Lytic bacteriophages specific to Flavobacterium columnare rescue catfish, Clarias batrachus (Linn.) from columnaris disease.

PubMed

Prasad, Yogendra; Arpana; Kumar, Dinesh; Sharma, A K

2011-03-01

This investigation was aimed to find out appropriate strategy against antibiotic resistant bacterial fish pathogen, F. columnare. This pathogen was found persistently associated with fishes causing columnaris disease and ensuing mass mortality in hatchery and culture system of Sub - Himalayan region. Nine lytic F. columnare phages (FCP1 - FCP9) specific to its fifteen isolates were isolated from the water and bottom sediments of various geo-climatic regions of North India. The F. columnare phage FCP1 (made of hexagonal head and non contractile long tail belonging to family Podovariedae, a member of DNA virus) exhibited broader host range to lyse 9 out of 15 isolates of F. columnare. Therapeutic ability of FCP1 phage was assessed in C. batrachus inoculated intramuscularly (im) with virulent bacterial isolate FC8 and post inoculated (PI) with FCP1 phage (@ 10(8) : 10(6):: cfu : pfu) through intramuscular (im), immersion (bath) and oral (phage impregnated feed) treatment. Significant (p < 0.001) reduction (less than 10(-3) cfu ml(-1)) in host bacterium in the sera, gill, liver and kidney of challenged fishes was noted after 6 hr of phage treatment. Quantum of phage played a significant role in bringing down bacterial population as in the sera of dose 1 (@ 4.55 x 10(6) pfu ml(-1)) and dose 2 (@ 9.15 x 10(6) pfu ml(-1)) treated fishes mean log10 cfu value reduced by 3 logs (58.39%) and 5 logs (73.77%) at 96 hr, respectively. Phage treatment led to disappearance of gross symptoms, negative bacteriological test, detectable phage and 100% survival in experimentally infected C. batrachus. Result of this study provides evidence of profound lytic impact of FCP1 phage and represents its interesting therapeutic importance against antibiotic resistant F. columnare.

Photodynamic inactivation of foodborne bacteria by eosin Y.

PubMed

Bonin, E; Dos Santos, A R; Fiori da Silva, A; Ribeiro, L H; Favero, M E; Campanerut-Sá, P A Z; de Freitas, C F; Caetano, W; Hioka, N; Mikcha, J M G

2018-06-01

The aim of this study was evaluate the effect of photodynamic inactivation mediated by eosin Y in Salmonella enterica serotype Typhimurium ATCC 14028, Escherichia coli ATCC 25922, Pseudomonas aeruginosa ATCC 27853, Staphylococcus aureus ATCC 25923 and Bacillus cereus ATCC 11778. Bacteria (10 7 CFU per ml) were incubated with eosin Y at concentrations ranging from 0·1 to 10 μmol l -1 , irradiated by green LED (λ max 490-570 nm) for 5, 10 and 15 min and the cellular viability was determined. Pseudomonas aeruginosa was completely inactivated when treated with 10 μmol l -1 eosin Y for 10 min. Treatments reduced B. cereus and Salm. Typhimurium counts to 2·7 log CFU per ml and 1·7 log CFU per ml, respectively. Escherichia coli counts were slightly reduced. Staphylococcus aureus presented the highest sensitivity, being completely inactivated by eosin Y at 5 μmol l -1 and 5 min of illumination. The reduction of cellular viability of photoinactivated Staph. aureus was also demonstrated by flow cytometry and morphological changes were observed by scanning electron microscopy. Eosin Y in combination with LED produced bacterial inactivation, being a potential candidate for photodynamic inactivation. This study evidenced the efficacy of photodynamic inactivation as a novel and promising alternative to bacterial control. © 2018 The Society for Applied Microbiology.

Efficacy of sanitizers in reducing Salmonella on pecan nutmeats during cracking and shelling.

PubMed

Beuchat, Larry R; Mann, David A; Alali, Walid Q

2013-05-01

Studies were done to evaluate the efficacy of chlorine (200 to 1,000 μg/ml), lactic acid (0.5 to 2%), levulinic acid (0.5 to 2%), sodium dodecyl sulfate (SDS, 0.05%), lactic acid plus SDS, levulinic acid plus SDS, and a mixed peroxyacid sanitizer (Tsunami 200, 40 and 80 μg/ml) in killing Salmonella on or in immersion- and on surface-inoculated pecan nutmeats (U.S. Department of Agriculture medium pieces and mammoth halves). The addition of SDS to treatment solutions containing lactic acid or levulinic acid resulted in generally higher reductions of Salmonella, but differences in these reductions were not always significant. Lactic and levulinic acids (2%) containing SDS (0.05%) were equivalent in killing Salmonella on immersion-inoculated nutmeats. Tsunami 200 (40 μg/ml) was less lethal or equivalent to 1 or 2% lactic and levulinic acids, with or without 0.05% SDS. Reductions did not exceed 1.1 log CFU/g of immersion-inoculated pieces and halves, regardless of sanitizer concentration or treatment time (up to 20 min). Reductions on surface-inoculated pieces and halves were 0.7 to 2.6 log CFU/g and 1.2 to 3.0 log CFU/g, respectively. Treatment with 2% lactic acid plus SDS (0.05%) and Tsunami (80 μg/ml) was most effective in killing Salmonella on surface-inoculated pieces; treatment of halves with chlorine (1,000 μg/ml) or lactic acid (1 or 2%), with or without SDS, was most efficacious. Exposure of immersion-inoculated pecan pieces to chlorine (200 μg/ml), lactic acid (2%) and levulinic acid (2%) with or without SDS, and Tsunami (80 μg/ml) during intermittent vacuum (18 ± 2 mbar) and ambient atmospheric pressure treatments for up to 20 min reduced Salmonella by only 0.1 to 1.0 log CFU/g. These studies emphasize the importance of preventing contamination of pecan nutmeats with Salmonella. Once nuts are contaminated, the lethality of sanitizers tested in this study is minimal.

Effectiveness of a Commercial Lactic Acid Bacteria Intervention Applied to Inhibit Shiga Toxin-Producing Escherichia coli on Refrigerated Vacuum-Aged Beef

PubMed Central

Kirsch, Katie R.; Tolen, Tamra N.; Hudson, Jessica C.; Griffin, Davey

2017-01-01

Because of their antagonistic activity towards pathogenic and spoilage bacteria, some members of the lactic acid bacteria (LAB) have been evaluated for use as food biopreservatives. The objectives of this study were to assess the antimicrobial utility of a commercial LAB intervention against O157 and non-O157 Shiga-toxigenic E. coli (STEC) on intact beef strip loins during refrigerated vacuum aging and determine intervention efficacy as a function of mode of intervention application. Prerigor strip loins were inoculated with a cocktail (8.9 ± 0.1 log10 CFU/ml) of rifampicin-resistant (100.0 μg/ml; RifR) O157 and non-O157 STEC. Inoculated loins were chilled to ≤4°C and treated with 8.7 ± 0.1 log10 CFU/ml LAB intervention using either a pressurized tank air sprayer (conventional application) or air-assisted electrostatic sprayer (ESS). Surviving STEC were enumerated on tryptic soy agar supplemented with 100.0 μg/ml rifampicin (TSAR) to determine STEC inhibition as a function of intervention application method (conventional, ESS) and refrigerated aging period (14, 28 days). Intervention application reduced STEC by 0.4 log10 CFU/cm2 (p < 0.05), although application method did not impact STEC reductions (p > 0.05). Data indicate that the LAB biopreservative may assist beef safety protection when utilized within a multi-intervention beef harvest, fabrication, and aging process. PMID:28630857

Bacteriophage application on red meats and poultry: Effects on Salmonella population in final ground products.

PubMed

Yeh, Y; Purushothaman, P; Gupta, N; Ragnone, M; Verma, S C; de Mello, A S

2017-05-01

This research was conducted to study the effects of bacteriophage application during tumbling on Salmonella populations in ground meat and poultry. Red meat trim and poultry were inoculated with a Salmonella cocktail to result in a contamination level of 7logCFU/g in ground products. A commercial preparation containing bacteriophages S16 and Felix-O1a (FO1a) was applied during tumbling at 10 7 and 10 8 PFU/ml. Samples were held at 4°C for 6h and 18h (red meat) and 30min and 6h (poultry). Overall, bacteriophage application on trim reduced 1 and 0.8logCFU/g of Salmonella in ground beef and ground pork, respectively. For ground chicken and ground turkey, Salmonella was reduced by 1.1 and 0.9logCFU/g, respectively. This study shows that bacteriophage application during tumbling of red meat trim and poultry can provide additional Salmonella control in ground products. Copyright © 2017 Elsevier Ltd. All rights reserved.

Early bactericidal activity of delamanid (OPC-67683) in smear-positive pulmonary tuberculosis patients.

PubMed

Diacon, A H; Dawson, R; Hanekom, M; Narunsky, K; Venter, A; Hittel, N; Geiter, L J; Wells, C D; Paccaly, A J; Donald, P R

2011-07-01

Delamanid (OPC-67683) is a novel mycolic acid biosynthesis inhibitor active against Mycobacterium tuberculosis at a low minimum inhibitory concentration. Forty-eight patients with smear-positive tuberculosis (63% male; 54.7 ± 9.9 kg; 30.7 ± 10.8 years) were randomly assigned to receive delamanid 100, 200, 300 or 400 mg daily for 14 days. Colony forming units (cfu) of M. tuberculosis were counted on agar plates from overnight sputum collections to calculate early bactericidal activity (EBA), defined as fall in log(10) cfu/ml sputum/day. The EBA of delamanid was monophasic and not significantly different between dosages; however, more patients receiving 200 mg (70%) and 300 mg (80%) experienced a response of ≥0.9 log(10) cfu/ml sputum decline over 14 days than those receiving 100 mg (45%) and 400 mg (27%). The average EBA of all dosages combined (0.040 ± 0.056 log(10) cfu/ml sputum/day) was significant from day 2 onward. Delamanid exposure was less than dosage-proportional, reaching a plateau at 300 mg, likely due to dose-limited absorption. Moderate but significant correlation was found between C(max) and EBA, indicating exposure dependence. Delamanid was well tolerated without significant toxicity. Delamanid at all dosages was safe, well tolerated and demonstrated significant exposure-dependent EBA over 14 days, supporting further investigation of its pharmacokinetics and anti-tuberculosis activity.

Microbiological profile and incidence of Salmonella and Listeria monocytogenes on hydroponic bell peppers and greenhouse cultivation environment.

PubMed

Avila-Vega, Dulce E; Alvarez-Mayorga, Beatriz; Arvizu-Medrano, Sofía M; Pacheco-Aguilar, Ramiro; Martínez-Peniche, Ramón; Hernández-Iturriaga, Montserrat

2014-11-01

The aim of this study was to generate information regarding the microbiological profile, including Salmonella and Listeria monocytogenes incidence, of hydroponically grown bell peppers and materials associated with their production in greenhouses located in Mexico. Samples of coconut fiber (24), knives (30), drippers (20), conveyor belts (161), pepper transportation wagons (30), air (178), water (16), nutrient solution for plant irrigation (78), and bell pepper fruits (528) were collected during one cycle of production (2009 to 2010) for the quantification of microbial indicators (aerobic plate counts [APC], molds, coliforms, and Escherichia coli) and the detection of Salmonella and L. monocytogenes. With regard to surfaces (conveyor belts and wagons) and utensils (knives and drippers), the APC, coliform, and mold counts ranged from 3.0 to 6.0, from 1.4 to 6.3, and from 3.6 to 5.2 log CFU/100 cm(2) or per utensil, respectively. The air in the greenhouse contained low median levels of APC (1.2 to 1.4 log CFU/100 liters) and molds (2.2 to 2.5 log CFU/100 liters). The median content of APC and coliforms in water were 0.5 log CFU/ml and 0.3 log MPN/100 ml, respectively. The median content of coliforms in nutrient solution ranged from 1.8 to 2.4 log MPN/100 ml, and E. coli was detected in 18 samples (range, <0.3 to 1.2 log MPN/100 ml). On bell pepper analyzed during the study, populations (median) of APC, coliforms, and molds were 5.4, 3.6, and 5.8 log CFU per fruit, respectively; E. coli was detected in 5.1% of the samples (range, 0.23 to 1.4 log MPN per fruit). Salmonella was isolated from only one sample (1.6%) of conveyor belt located at the packing area and in four bell pepper samples (3%). L. monocytogenes was not detected. This information could help producers to establish effective control measures to prevent the presence of foodborne pathogens in bell peppers based on a scientific approach.

Liquid egg white pasteurization using a centrifugal UV irradiator.

PubMed

Geveke, David J; Torres, Daniel

2013-03-01

Studies are limited on UV nonthermal pasteurization of liquid egg white (LEW). The objective of this study was to inactivate Escherichia coli using a UV irradiator that centrifugally formed a thin film of LEW on the inside of a rotating cylinder. The LEW was inoculated with E. coli K12 to approximately 8 log cfu/ml and was processed at the following conditions: UV intensity 1.5 to 9.0 mW/cm²; cylinder rotational speed 450 to 750 RPM, cylinder inclination angle 15° to 45°, and flow rate 300 to 900 ml/min, and treatment time 1.1 to 3.2s. Appropriate dilutions of the samples were pourplated with tryptic soy agar (TSA). Sublethal injury was determined using TSA+4% NaCl. The regrowth of surviving E. coli during refrigerated storage for 28 days was investigated. The electrical energy of the UV process was also determined. The results demonstrated that UV processing of LEW at a dose of 29 mJ/cm² at 10°C reduced E. coli by 5 log cfu/ml. Inactivation significantly increased with increasing UV dose and decreasing flow rate. The results at cylinder inclination angles of 30° and 45° were similar and were significantly better than those at 15°. The cylinder rotational speed had no significant effect on inactivation. The occurrence of sublethal injury was detected. Storage of UV processed LEW at 4° and 10°C for 21 days further reduced the population of E. coli to approximately 1 log cfu/ml where it remained for an additional 7 days. The UV energy applied to the LEW to obtain a 5 log reduction of E. coli was 3.9 J/ml. These results suggest that LEW may be efficiently pasteurized, albeit at low flow rates, using a nonthermal UV device that centrifugally forms a thin film. Published by Elsevier B.V.

Short chain nitrocompounds as a treatment of layer hen manure and litter; effects on in vitro survivability of Salmonella, generic E. coli and nitrogen metabolism.

PubMed

Ruiz-Barrera, Oscar; Anderson, Robin C; Hume, Michael E; Corrales-Millan, Jonatan; Castillo-Castillo, Yamicela; Corral-Luna, Agustin; Guevara-Valdez, Jose Luis; Salinas-Chavira, Jaime; Rodriguez-Muela, Carlos; Arzola-Alvarez, Claudio

2017-01-02

The current study was conducted to assess the bactericidal effectiveness of several nitrocompounds against pathogens in layer hen manure and litter. Evidence from an initial study indicated that treatment of layer hen manure with 12 mM nitroethane decreased populations of generic E. coli and total coliforms by 0.7 and 2.2 log 10 colony forming units (CFU) g -1 , respectively, after 24 h aerobic incubation at ambient temperature when compared to untreated populations. Salmonella concentrations were unaffected by nitroethane in this study. In a follow-up experiment, treatment of 6-month-old layer hen litter (mixed with 0.4 mL water g -1 ) with 44 mM 2-nitroethanol, 2-nitropropanol or ethyl nitroacetate decreased an inoculated Salmonella typhimurium strain from its initial concentration (3 log 10 CFU g -1 ) by 0.7 to 1.7 log 10 CFU g -1 after 6 h incubation at 37°C in covered containers. After 24 h incubation, populations of the inoculated S. Typhmiurium in litter treated with 44 mM 2-nitroethanol, 2-nitropropanol, ethyl nitroacetate or nitroethane were decreased more than 3.2 log 10 CFU g -1 compared to populations in untreated control litter. Treatment of litter with 44 mM 2-nitroethanol, 2-nitropropanol, ethyl nitroacetate decreased rates of ammonia accumulation more than 70% compared to untreated controls (0.167 µmol mL -1 h -1 ) and loses of uric acid (< 1 µmol mL -1 ) were observed only in litter treated with 44 mM 2-nitropropanol, indicating that some of these nitrocompounds may help prevent loss of nitrogen in treated litter. Results warrant further research to determine if these nitrocompounds can be developed into an environmentally sustainable and safe strategy to eliminate pathogens from poultry litter, while preserving its nitrogen content as a nutritionally valuable crude protein source for ruminants.

Comparative Activities of Ciprofloxacin and Levofloxacin against Streptococcus pneumoniae in an In Vitro Dynamic Model

PubMed Central

Zinner, Stephen H.; Simmons, Kelly; Gilbert, Deborah

2000-01-01

The activities of levofloxacin (500 mg every 24 h) and ciprofloxacin (750 mg every 12 h) against six pneumococcal isolates in an in vitro dynamic model were compared. For one strain, levofloxacin reduced the inoculum by over 4 log CFU/ml and ciprofloxacin reduced the inoculum by over 2 log CFU/ml. For four isolates, both drugs reduced inocula by 4 log CFU/ml within 6 h, suggesting that this dose of ciprofloxacin should be as effective as levofloxacin against these pneumococci. PMID:10681356

Monitoring of microbial hazards at farms, slaughterhouses, and processing lines of swine in Korea.

PubMed

Rho, M J; Chung, M S; Lee, J H; Park, J

2001-09-01

This study was executed to investigate microbiological hazards at swine farms, slaughterhouses, dressing operations, and local markets for the application of the hazard analysis critical control point system in Korea by analyzing total aerobic plate count (APC) and presence of pathogens. Six integrated pig farms and meat packers were selected from six different provinces, and samples were collected from pig carcasses by swabbing and excision methods at the slaughterhouses, processing rooms, and local markets, respectively. APCs of water in water tanks were relatively low, 1.9 to 3.1 log10 CFU/ml; however, they were increased to 4.6 to 6.9 log10 CFU/ml when sampled from water nipples in the pigpen. APCs of feeds in the feed bins and in the pigpens were 4.4 to 5.4 and 5.2 to 6.7 log10 CFU/g, respectively. Salmonella spp., Staphylococcus aureus, and Clostridium perfringens were detected from water and feed sampled in pigpens and pigpen floors. S. aureus was the most frequently detected pathogenic bacteria in slaughterhouses and processing rooms. Listeria monocytogenes and Yersinia enterocolitica were also detected from the processing rooms of the Kyonggi, Kyongsang, and Cheju provinces. Even though APCs were maintained at the low level of 3.0 log10 CFU/g during slaughtering and processing steps, those of final pork products produced by the same companies showed relatively high numbers when purchased from the local market. These results indicated that the cold chain system for transporting and merchandising of pork products was deficient in Korea. Water supply and feed bins in swine farms and individual operations can be identified as critical control points to reduce microbiological hazards in swine farms, slaughterhouses, and processing plants.

Activities of tigecycline (GAR-936) against Legionella pneumophila in vitro and in guinea pigs with L. pneumophila pneumonia.

PubMed

Edelstein, Paul H; Weiss, William J; Edelstein, Martha A C

2003-02-01

The activities of tigecycline (Wyeth Research) against extracellular and intracellular Legionella pneumophila and for the treatment of guinea pigs with L. pneumophila pneumonia were studied. The tigecycline MIC at which 50% of strains are inhibited for 101 different Legionella sp. strains was 4 micro g/ml versus 0.125 and 0.25 micro g/ml for azithromycin and erythromycin, respectively. Tigecycline was about as active as erythromycin (tested at 1 micro g/ml) against the F889 strain of L. pneumophila grown in guinea pig alveolar macrophages and more active than erythromycin against the F2111 strain. Azithromycin (0.25 micro g/ml) was more active than (F889) or as active as (F2111) tigecycline (1 micro g/ml) in the macrophage model. When tigecycline was given (7.5 mg/kg of body weight subcutaneously once) to guinea pigs with L. pneumophila pneumonia, the mean peak serum and lung levels were 2.3 and 1.8 micro g/ml (1.2 and 1.5 micro g/g) at 1 and 2 h postinjection, respectively. The serum and lung areas under the concentration time curve from 0 to 24 h were 13.7 and 15.8 micro g. h/ml, respectively. Thirteen of 16 guinea pigs with L. pneumophila pneumonia treated with tigecycline (7.5 mg/kg subcutaneously once daily for 5 days) survived for 7 days post-antimicrobial therapy, as did 11 of 12 guinea pigs treated with azithromycin (15 mg/kg intraperitoneally once daily for 2 days). None of 12 guinea pigs treated with saline survived. Tigecycline-treated guinea pigs had average end of therapy lung counts of 1 x 10(6) CFU/g (range, 2.5 x 10(4) to 3.2 x 10(6) CFU/g) versus <1 x 10(2) CFU/g for azithromycin (range, undetectable to 100 CFU/g). A second guinea pig study examined the ability of tigecycline to clear L. pneumophila from the lung after 5 to 9 days of therapy; bacterial concentrations 1 day posttherapy ranged from log(10) 4.2 to log(10) 5.5 CFU/g for four different dosing regimens. Tigecycline is about as effective as erythromycin against intracellular L. pneumophila, but tigecycline inactivation by the test media confounded the interpretation of susceptibility data. Tigecycline was effective at preventing death from pneumonia in an animal model of Legionnaires' disease, warranting human clinical trials of the drug for the disease.

Antimicrobial activity of buttermilk and lactoferrin peptide extracts on poultry pathogens.

PubMed

Jean, Catherine; Boulianne, Martine; Britten, Michel; Robitaille, Gilles

2016-11-01

Antibiotics are commonly used in poultry feed as growth promoters. This practice is questioned given the arising importance of antibiotic resistance. Antimicrobial peptides can be used as food additives for a potent alternative to synthetic or semi-synthetic antibiotics. The objective of this study was to develop a peptide production method based on membrane adsorption chromatography in order to produce extracts with antimicrobial activity against avian pathogens (Salmonella enterica var. Enteritidis, Salmonella enterica var. Typhimurium, and two Escherichia coli strains, O78:H80 and TK3 O1:K1) as well as Staphylococcus aureus. To achieve this, buttermilk powder and purified lactoferrin were digested with pepsin. The peptide extracts (<10 kDa) were fractionated depending on their charges through high-capacity cation-exchange and anion-exchange adsorptive membranes. The yields of cationic peptide extracts were 6·3 and 15·4% from buttermilk and lactoferrin total peptide extracts, respectively. Antimicrobial activity was assessed using the microdilution technique on microplates. Our results indicate that the buttermilk cationic peptide extracts were bactericidal at less than 5 mg/ml against the selected avian strains, with losses of 1·7 log CFU/ml (Salm. Typhimurium) to 3 log CFU/ml (E. coli O78:H80); viability decreased by 1·5 log CFU/ml for Staph. aureus, a Gram-positive bacterium. Anionic and non-adsorbed peptide extracts were inactive at 5 mg/ml. These results demonstrate that membrane adsorption chromatography is an effective way to prepare a cationic peptide extract from buttermilk that is active against avian pathogens.

Determination of serum bactericidal activity against Escherichia coli by an automated photometric method.

PubMed Central

Crokaert, F; Lismont, M J; van der Linden, M P; Yourassowsky, E

1988-01-01

The resistance of gram-negative bacteria to complement-mediated serum activity is supposedly an important virulence factor. However, the lack of standardization in the methods used to determine serum activity and the many definitions applied make the comparisons between studies very difficult. We developed a rapid photometric method that we compared with a classical killing one. Escherichia coli in the exponential phase of growth in brain heart infusion broth (final inoculum, 10(7) CFU/ml) at 35 degrees C was added to 50% human serum in Veronal buffer. Viable counts and automatic recording of the variations in the optical densities were obtained for 40 E. coli strains isolated from the stools of healthy adults. With the viable count method, 17 (42.5%) were susceptible (at least a 1 log CFU/ml decrease), 17 (42.5%) were resistant (a 0.6 log CFU/ml increase), 4 (10%) were intermediate (poorly growing inoculum or a decrease of less than 1 log CFU/ml), and 2 could not be classified (nonreproducible results). Agreement between both methods was observed for 87.5% of the stool strains. Eight reference strains of known susceptibilities were classified identically by both methods, leading to a final concordance rate of 89.6%. A total of 129 blood isolates were tested by the photometric method: 64 (49.6%) were resistant, 50 (38.8%) were susceptible 5 (3.9%) showed early regrowth, and 10 (7.7%) were not perfectly reproducible. Of these 129 blood isolates, 5 were also tested by the killing method: 37 (49%) were resistant, 32 (43%) were susceptible, and 6 (8%) were intermediate. The concordance rate between both assays was 89% for the blood isolates; when the minor discordances were ruled out, it was 97%. This automated method could be a useful screening tool for detecting resistance to serum in clinical trials and for studying the in vitro variations of this property. PMID:3053761

Impact of accessory gene regulator (agr) dysfunction on vancomycin pharmacodynamics among Canadian community and health-care associated methicillin-resistant Staphylococcus aureus.

PubMed

Tsuji, Brian T; MacLean, Robert D; Dresser, Linda D; McGavin, Martin J; Simor, Andrew E

2011-05-20

The accessory gene regulator (agr) is a quorum sensing cluster of genes which control colonization and virulence in Staphylococcus aureus. We evaluated agr function in community- (CA) and healthcare-associated (HA) MRSA, to compare the pharmacodynamics and bactericidal activity of vancomycin against agr functional and dysfunctional HA-MRSA and CA-MRSA. 40 clinical isolates of MRSA from the Canadian Nosocomial Infection Surveillance Program were evaluated for delta-haemolysin production, as a surrogate marker of agr function. Time kill experiments were performed for vancomycin at 0 to 64 times the MIC against an initial inoculum of 10(6) and 10(8) cfu/ml of agr functional and dysfunctional CA-MRSA and HA-MRSA and these data were fit to a hill-type pharmacodynamic model. 15% isolates were agr dysfunctional, which was higher among HA-MRSA (26.3%) versus CA-MRSA (4.76%). Against a low initial inoculum of 10(6) cfu/ml of CA-MRSA, vancomycin pharmacodynamics were similar among agr functional and dysfunctional strains. However, against a high initial inoculum of 10(8) cfu/ml, killing activity was notably attenuated against agr dysfunctional CA-MRSA (USA400) and HA-MRSA (USA100). CA-MRSA displayed a 20.0 fold decrease in the maximal reduction in bacterial counts (Emax) which was 3.71 log(10) CFU/ml for agr functional vs. 2.41 log(10) CFU/ml for agr dysfunctional MRSA (p = 0.0007). Dysfunction in agr was less common among CA-MRSA vs. HA-MRSA. agr dysfunction demonstrated an impact on vancomycin bactericidal activity and pharmacodynamics against a high initial inoculum of CA-MRSA and HA-MRSA, which may have implications for optimal antimicrobial therapy against persistent, difficult to treat MRSA infections.

Efficacy of chlorine dioxide against Listeria monocytogenes in brine chilling solutions.

PubMed

Valderrama, W B; Mills, E W; Cutter, C N

2009-11-01

Chilled brine solutions are used by the food industry to rapidly cool ready-to-eat meat products after cooking and before packaging. Chlorine dioxide (ClO(2)) was investigated as an antimicrobial additive to eliminate Listeria monocytogenes. Several experiments were performed using brine solutions made of sodium chloride (NaCl) and calcium chloride (CaCl(2)) inoculated with L. monocytogenes and/or treated with 3 ppm of ClO(2). First, 10 and 20% CaCl(2) and NaCl solutions (pH 7.0) were inoculated with a five-strain cocktail of L. monocytogenes to obtain approximately 7 log CFU/ml and incubated 8 h at 0 degrees C. The results demonstrated that L. monocytogenes survived in 10% CaCl(2), 10 and 20% NaCl, and pure water. L. monocytogenes levels were reduced approximately 1.2 log CFU/ml in 20% CaCl(2). Second, inoculated ( approximately 7 log CFU/ml) brine solutions (10 and 20% NaCl and 10% CaCl(2)) treated with 3 ppm of ClO(2) resulted in a approximately 4-log reduction of the pathogen within 90 s. The same was not observed in a solution of 20% CaCl(2); further investigation demonstrated that high levels of divalent cations interfere with the disinfectant. Spent brine solutions from hot dog and ham chilling were treated with ClO(2) at concentrations of 3 or 30 ppm. At these concentrations, ClO(2) did not reduce L. monocytogenes. Removal of divalent cations and organic material in brine solutions prior to disinfection with ClO(2) should be investigated to improve the efficacy of the compound against L. monocytogenes. The information from this study may be useful to processing establishments and researchers who are investigating antimicrobials in chilling brine solutions.

The antibacterial effect of Ziziphora clinopodioides essential oil and nisinagainst Salmonella typhimurium and Staphylococcus aureus in doogh, a yoghurt-based Iranian drink.

PubMed

Shahbazi, Yasser

2016-01-01

Doogh is the most popular and commonly consumed yoghurt-based Iranian drink. The aim of this study was to investigate the antibacterial effects of Ziziphora clinopodioides essential oil (ZEO) at 0.10 and 0.20% concentrations, nisin at 250 and 500 IU mL -1 , and their combination against Salmonella typhimurium and Staphylococcus aureus in doogh during storage at 4 ˚C for  9 days. Nine batches were studied as follows: control: no ZEO or nisin added, A: 0.10% ZEO, B: 0.20% ZEO, C: 250 IU mL -1 nisin, D: 500 IU mL -1 nisin, E: 0.10% ZEO + 250 IU mL -1 nisin, F: 0.10% ZEO + 500 IU mL -1 nisin, G: 0.20% ZEO + 250 IU mL -1 nisin and H: 0.20% ZEO + 500 IU mL -1 nisin. Based on gas-chromatography and mass spectrometry, carvacrol (65.22%), thymol (19.51%), p -cymene (4.86%) and ɣ -terpinene (4.63%) were the major components of ZEO. The populations of S. typhimurium and S. aureus in samples treated with all concentrations of the ZEO and nisin were kept below 1 log CFU mL -1 on day 5 of storage, while the count of S . typhimurium and S. aureus was found as 2.72 ± 0.02 and 2.21 ± 0.00 log CFU mL -1 on day 5 for untreated samples, respectively. The ZEO separately and in combination with nisin, was very effective against these two common food-borne pathogens. The ZEO alone and in combination with nisin could be considered as a potential strong antimicrobial agent that can be used for the growth inhibition of aforementioned bacteria in food products especially doogh.

Cronobacter sakazakii reduction by blueberry proanthocyanidins.

PubMed

Joshi, Snehal S; Howell, Amy B; D'Souza, Doris H

2014-05-01

Blueberry juice and blueberry polyphenols reportedly have antimicrobial properties against foodborne pathogens, without much currently known on their effects against Cronobacter sakazakii. This study evaluated the antimicrobial effects of blueberry proanthocyanidins (PAC) and commercial blueberry juice (BJ) against two strains of C. sakazakii, ATCC 29004 and 29544. BJ (pH 2.8), blueberry PAC (5 mg/ml) and controls (phosphate buffered saline (PBS), pH 7.2, and malic acid pH 3.0) were mixed with equal volumes of washed overnight cultures of C. sakazakii and incubated for 30 min, 1 h, 3 h and 6 h at 37°C. Reductions of ∼1 and 1.50 log CFU/ml were obtained for strains 29004 and 29544, respectively after 30 min with BJ or blueberry PAC. Both C. sakazakii strains 29004 and 29544 were reduced to undetectable levels from 8.25 ± 0.12 log CFU/ml and 8.48 ± 0.03 log CFU/ml, respectively with BJ (pH 2.8) or blueberry PAC after 1 h, while malic acid (pH 3.0) showed ∼1.3 log CFU/ml reduction for both strains. Scanning electron microscopy studies showed differences in cell membrane morphology with clumping and formation of blebs of the treated strains compared to untreated controls. These results warrant further in vivo studies with blueberry bioactives to determine potential for preventing and treating C. sakazakii infections. Copyright © 2013. Published by Elsevier Ltd.

Disinfection alternatives for contact surfaces and toys at child care centers.

PubMed

Jimenez, Maribel; Martinez, Celida I; Chaidez, Cristobal

2010-12-01

Child care surfaces are vehicles for disease-causing organisms. Disinfectant procedures prevent microbial dispersion. This study reports the effectiveness of CITRUS Farm Edition® (CFE), Clorox® GreenWorks™ (CGW) and Clorox® Anywhere® (CA) against Salmonella Typhimurium and Staphylococcus aureus inoculated (1 ml of 9Log(10) CFU/ml) on a high chair and ball toy. Disinfectants were sprayed and bacteria recovered from surfaces by sponge method. Exposing an inoculated high chair to CA resulted in the highest reduction of S. aureus (3.92 Log(10)) and S. Typhimurium (3.22 Log(10)). CGW reduced S. aureus and S. Typhimurium by 2.84 and 2.12 Log(10) from the inoculated high chair, while the inoculated ball toy showed a 2.50 and 1.80 Log(10) reduction, respectively. CFE showed the lowest reduction with 1.42 and 1.53 Log(10) of S. aureus and S. Typhimurium from the inoculated ball toy. CA was the best disinfectant no matter which bacteria or surface was analyzed. Emphasis on the effectiveness of disinfectant products is needed to be included in child care center infection control programs.

Effect of lecithin and starch on alginate-encapsulated probiotic bacteria.

PubMed

Donthidi, A R; Tester, R F; Aidoo, K E

2010-01-01

The effect of lecithin and starch on viability of alginate encapsulated probiotics was determined at different temperatures. Probiotic organisms (1% v/v>10Log CFU ml(-1)) were encapsulated using alginate (2% w/v), gelatinized starches (2% w/v) and lecithin (0-4% w/v) and stored in sealed containers at 4, 23 and 37 degrees C (to simulate shelf storage conditions). Incorporation of lecithin improved the entrapment efficiency (p < 0.05) and the viability of encapsulated bacteria (p = 0.02). Encapsulated Lactobacillus, Bifidobacterium species and Lactococcus lactis in lecithin containing freeze-dried beads had good survival stability (above 6Log CFU ml(-1)) at 23 degrees C for 12 weeks. The bacteria in the beads showed 6Log survival by the end of 2 weeks at 37 degrees C. Encapsulated L. casei in the alginate beads containing lecithin were also more stable in the yoghurt than the beads without lecithin. SEM analysis of the beads showed an irregular surface for the beads without lecithin.

Growth and enterotoxin production of Staphylococcus aureus during the manufacture and ripening of Camembert-type cheeses from raw goats' milk.

PubMed

Meyrand, A; Boutrand-Loei, S; Ray-Gueniot, S; Mazuy, C; Gaspard, C E; Jaubert, G; Perrin, G; Lapeyre, C; Vernozy-Rozand, C

1998-09-01

Tests were carried out to determine the effect of manufacturing procedures for a Camembert-type cheese from raw goats' milk on the growth and survival of Staphylococcus aureus organisms added to milk at the start of the process, and to study the possible presence of staphylococcal enterotoxin A in these cheeses. The initial staphylococcal counts were, respectively, 2, 3, 4, 5 and 6 log cfu ml-1. Cheese was prepared following the industrial specifications and ripened for 41 d. Detection of enterotoxins was done by the Vidas SET test and by an indirect double-sandwich ELISA technique using antienterotoxin monoclonal antibodies. Generally, numbers of microbes increased at a similar rate during manufacture in all cheeses until salting. During the ripening period, the aerobic plate count population and Staph. aureus levels remained stable and high. There was an approximately 1 log reduction of Staph. aureus in cheeses made with an initial inoculum of Staph. aureus greater than 10(3) cfu ml-1 at the end of the ripening period (41 d) compared with the count at 22 h. The level of staphylococcal enterotoxin A recovered varied from 1 to 3.2 ng g-1 of cheese made with an initial population of 10(3)-10(6) cfu ml-1. No trace of enterotoxin A was detected in cheeses made with the lowest Staph. aureus inoculum used in this study.

Effects of bilimbi (Averrhoa bilimbi L.) and tamarind (Tamarindus indica L.) juice on Listeria monocytogenes Scott A and Salmonella Typhimurium ATCC 14028 and the sensory properties of raw shrimps.

PubMed

Norhana, M N Wan; Azman, Mohd Nor A; Poole, Susan E; Deeth, Hilton C; Dykes, Gary A

2009-11-30

The potential of using juice of bilimbi (Averrhoa bilimbi L.) and tamarind (Tamarindus indica L.) to reduce Listeria monocytogenes Scott A and Salmonella Typhimurium ATCC 14028 populations on raw shrimps after washing and during storage (4 degrees C) was investigated. The uninoculated raw shrimps and those inoculated with approximately 9 log cfu/ml of L. monocytogenes Scott A and S. Typhimurium ATCC 14028 were washed (dipped or rubbed) in distilled water (SDW) (control), bilimbi or tamarind juice at 1:4 (w/v) concentrations for 10 and 5 min. Naturally occurring aerobic bacteria (APC), L. monocytogenes Scott A and S. Typhimurium ATCC 14028 counts, pH values and sensory analysis of washed shrimps were determined immediately after washing (day 0), and on days 3 and 7 of storage. Compared to SDW, bilimbi and tamarind juice significantly (p<0.05) reduced APC (0.40-0.70 log cfu/g), L. monocytogenes Scott A (0.84-1.58 log cfu/g) and S. Typhimurium ATCC 14028 (1.03-2.00 log cfu/g) populations immediately after washing (0 day). There was a significant difference (p<0.05) in bacterial reduction between the dipping (0.40-0.41 log for APC; 0.84 for L. monocytogenes Scott A and 1.03-1.09 log for S. Typhimurium ATCC 14028) and rubbing (0.68-0.70 log for APC; 1.34-1.58 for L. monocytogenes Scott A and 1.67-2.00 log for S. Typhimurium ATCC 14028) methods. Regardless of washing treatments or methods, populations of S. Typhimurium ATCC 14028 decreased slightly (5.10-6.29 log cfu/g on day 7 of storage) while populations of L. monocytogenes Scott A (8.74-9.20 log cfu/g) and APC (8.68-8.92 log cfu/g) increased significantly during refrigerated storage. The pH of experimental shrimps were significantly (p<0.05) decreased by 0.15-0.22 pH units after washing with bilimbi and tamarind juice. The control, bilimbi or tamarind-washed shrimps did not differ in sensory panellist acceptability (p>0.05) throughout the storage except for odour (p<0.05) attributes at 0 day when acidic or lemony smell was noticed in bilimbi- and tamarind-washed shrimps and not in control shrimps.

Effect of high-temperature, short-time (HTST) pasteurization on milk containing low numbers of Mycobacterium paratuberculosis.

PubMed

Grant, I R; Ball, H J; Rowe, M T

1998-02-01

The efficacy of high-temperature, short-time (HTST) pasteurization (72 degrees C/15 s) when low numbers (< or = 10(3) cfu ml-1) of Mycobacterium paratuberculosis are present in milk was investigated. Raw cows' milk spiked with Myco. paratuberculosis (10(3) cfu ml-1, 10(2) cfu ml-1, 10 cfu ml-1, and 10 cfu 50 ml-1) was subjected to HTST pasteurization using laboratory pasteurizing units. Ten bovine strains of Myco. paratuberculosis were tested in triplicate. Culture in BACTEC Middlebrook 12B radiometric medium detected acid-fast survivors in 14.8% and 10% of HTST-pasteurized milk samples at the 10(3) and 10(2) cfu ml-1 inoculum levels, respectively, whereas conventional culture on Herrold's egg yolk medium containing mycobactin J detected acid-fast survivors in only 3.7% and 6.7% of the same milk samples. IS900-based PCR confirmed that these acid-fast survivors were Myco. paratuberculosis. No viable Myco. paratuberculosis were isolated from HTST-pasteurized milk initially containing either 10 cfu ml-1 or 10 cfu 50 ml-1.

Heat Resistance Mediated by pLM58 Plasmid-Borne ClpL in Listeria monocytogenes

PubMed Central

Aalto-Araneda, Mariella; Lindström, Miia; Korkeala, Hannu

2017-01-01

ABSTRACT Listeria monocytogenes is one of the most heat-resistant non-spore-forming food-borne pathogens and poses a notable risk to food safety, particularly when mild heat treatments are used in food processing and preparation. While general heat stress properties and response mechanisms of L. monocytogenes have been described, accessory mechanisms providing particular L. monocytogenes strains with the advantage of enhanced heat resistance are unknown. Here, we report plasmid-mediated heat resistance of L. monocytogenes for the first time. This resistance is mediated by the ATP-dependent protease ClpL. We tested the survival of two wild-type L. monocytogenes strains—both of serotype 1/2c, sequence type ST9, and high sequence identity—at high temperatures and compared their genome composition in order to identify genetic mechanisms involved in their heat survival phenotype. L. monocytogenes AT3E was more heat resistant (0.0 CFU/ml log10 reduction) than strain AL4E (1.4 CFU/ml log10 reduction) after heating at 55°C for 40 min. A prominent difference in the genome compositions of the two strains was a 58-kb plasmid (pLM58) harbored by the heat-resistant AT3E strain, suggesting plasmid-mediated heat resistance. Indeed, plasmid curing resulted in significantly decreased heat resistance (1.1 CFU/ml log10 reduction) at 55°C. pLM58 harbored a 2,115-bp open reading frame annotated as an ATP-dependent protease (ClpL)-encoding clpL gene. Introducing the clpL gene into a natively heat-sensitive L. monocytogenes strain (1.2 CFU/ml log10 reduction) significantly increased the heat resistance of the recipient strain (0.4 CFU/ml log10 reduction) at 55°C. Plasmid-borne ClpL is thus a potential predictor of elevated heat resistance in L. monocytogenes. IMPORTANCE Listeria monocytogenes is a dangerous food pathogen causing the severe illness listeriosis that has a high mortality rate in immunocompromised individuals. Although destroyed by pasteurization, L. monocytogenes is among the most heat-resistant non-spore-forming bacteria. This poses a risk to food safety, as listeriosis is commonly associated with ready-to-eat foods that are consumed without thorough heating. However, L. monocytogenes strains differ in their ability to survive high temperatures, and comprehensive understanding of the genetic mechanisms underlying these differences is still limited. Whole-genome-sequence analysis and phenotypic characterization allowed us to identify a novel plasmid, designated pLM58, and a plasmid-borne ATP-dependent protease (ClpL), which mediated heat resistance in L. monocytogenes. As the first report on plasmid-mediated heat resistance in L. monocytogenes, our study sheds light on the accessory genetic mechanisms rendering certain L. monocytogenes strains particularly capable of surviving high temperatures—with plasmid-borne ClpL being a potential predictor of elevated heat resistance. PMID:29104933

Evaluation of lactic acid bacterium fermentation products and food-grade chemicals to control Listeria monocytogenes in blue crab (Callinectes sapidus) meat.

PubMed Central

Degnan, A J; Kaspar, C W; Otwell, W S; Tamplin, M L; Luchansky, J B

1994-01-01

Fresh blue crab (Callinectes sapidus) meat was obtained from retail markets in Florida and sampled for viable Listeria monocytogenes. The pathogen was found in crabmeat in three of four different lots tested by enrichment and at levels of 75 CFU/g in one of the same four lots by direct plating. Next, crabmeat was steam sterilized, inoculated with a three-strain mixture of L. monocytogenes (ca. 5.5 log10 CFU/g), washed with various lactic acid bacterium fermentation products (2,000 to 20,000 arbitrary units [AU]/ml of wash) or food-grade chemicals (0.25 to 4 M), and stored at 4 degrees C. Counts of the pathogen remained relatively constant in control samples during storage for 6 days, whereas in crabmeat washed with Perlac 1911 or MicroGard (10,000 to 20,000 AU), numbers initially decreased (0.5 to 1.0 log10 unit/g) but recovered to original levels within 6 days. Numbers of L. monocytogenes cells decreased 1.5 to 2.7 log10 units/g of crabmeat within 0.04 day when washed with 10,000 to 20,000 AU of Alta 2341, enterocin 1083, or Nisin per ml. Thereafter, counts increased 0.5 to 1.6 log10 units within 6 days. After washing with food-grade chemicals, modest reductions (0.4 to 0.8 log10 unit/g) were observed with sodium acetate (4 M), sodium diacetate (0.5 or 1 M), sodium lactate (1 M), or sodium nitrite (1.5 M). However, Listeria counts in crabmeat washed with 2 M sodium diacetate decreased 2.6 log10 units/g within 6 days. In addition, trisodium phosphate reduced L. monocytogenes counts from 1.7 (0.25 M) to > 4.6 (1 M) log10 units/g within 6 days.(ABSTRACT TRUNCATED AT 250 WORDS) PMID:7944362

Development of a quantitative PCR for detection of Lactobacillus plantarum starters during wine malolactic fermentation.

PubMed

Cho, Gyu-Sung; Krauss, Sabrina; Huch, Melanie; Du Toit, Maret; Franz, Charles M A P

2011-12-01

A quantitative, real-time PCR method was developed to enumerate Lactobacillus plantarum IWBT B 188 during the malolactic fermentation (MLF) in Grauburgunder wine. The qRT-PCR was strain-specific, as it was based on primers targeting a plasmid DNA sequence, or it was L. plantarum-specific, as it targeted a chromosomally located plantaricin gene sequence. Two 50 l wine fermentations were prepared. One was inoculated with 15 g/hl Saccharomyces cerevisiae, followed by L. plantarum IWBT B 188 at 3.6 × 10(6) CFU/ml, whereas the other was not inoculated (control). Viable cell counts were performed for up to 25 days on MRS agar, and the same cells were enumerated by qRT-PCR with both the plasmid or chromosomally encoded gene primers. The L. plantarum strain survived under the harsh conditions in the wine fermentation at levels above 10(5)/ml for approx. 10 days, after which cell numbers decreased to levels of 10(3) CFU/ml at day 25, and to below the detection limit after day 25. In the control, no lactic acid bacteria could be detected throughout the fermentation, with the exception of two sampling points where ca. 1 × 10(2) CFU/ml was detected. The minimum detection level for quantitative PCR in this study was 1 × 10(2) to 1 × 10(3) CFU/ml. The qRT-PCR results determined generally overestimated the plate count results by about 1 log unit, probably as a result of the presence of DNA from dead cells. Overall, qRT-PCR appeared to be well suited for specifically enumerating Lactobacillus plantarum starter cultures in the MLF in wine.

Inactivation of bacteria from contaminated streams in Limpopo, South Africa by silver- or copper-nanoparticle paper filters

PubMed Central

Dankovich, Theresa A.; Levine, Jonathan S.; Potgieter, Natasha; Dillingham, Rebecca; Smith, James A.

2016-01-01

There is an urgent need for inexpensive point-of-use methods to purify drinking water in developing countries to reduce the incidence of illnesses caused by waterborne pathogens. Previously, our work showed the deactivation of laboratory-cultured bacteria by percolation through a thick paper sheet containing either silver (Ag) or copper (Cu) nanoparticles (NP). In this study, these paper filters containing AgNPs or CuNPs have been tested with water sourced from contaminated streams in Limpopo, South Africa. Following the percolation of the contaminated stream water through the metal nanoparticle (MNP) papers, the water quality of the filtered effluent was evaluated with respect to the colony counts of total coliform and E. coli bacteria, turbidity, and either silver or copper ions. Influent total coliform bacteria concentrations from the stream water in Limpopo ranged from 250 CFU/100 mL to 1,750,000 CFU/100 mL. With the less contaminated stream water (250 - 15,000 CFU/100 mL), both AgNP and CuNP papers showed complete inactivation of the coliform bacteria. With the surface water with higher coliform bacteria levels (500,000 - 1,000,000 CFU/100 mL), both the AgNP and CuNP papers showed similar results with a slightly higher bacteria reduction of log10 5.1 for the AgNP papers than the log10 4.8 reduction for the CuNP papers. E. coli results followed similar trends. For most water purification experiments, the metal release from the sheets was minimal, with values under 0.1 ppm for Ag and 1.0 ppm for Cu (the current US EPA and WHO drinking water limits for Ag and Cu, respectively). These results show good potential for the use of paper embedded with silver and/or copper nanoparticles as effective point-of-use water purifiers. PMID:27022474

Changes in the microbial composition of raw milk induced by thermization treatments applied prior to traditional Greek hard cheese processing.

PubMed

Samelis, John; Lianou, Alexandra; Kakouri, Athanasia; Delbès, Céline; Rogelj, Irena; Bogovic-Matijasić, Bojana; Montel, Marie-Christine

2009-04-01

The microbiological quality, safety, and composition of mixtures of ewe's and goat's milk (90:10) used for cheesemaking were evaluated before and after thermization at 60 and 67 degrees C for 30 s. Such mild thermal treatments are commonly applied to reduce natural contaminants of raw milk before processing for traditional hard Greek cheeses. Raw milk samples had an average total bacterial count of 7.3 log CFU/ml; most of these bacteria were lactic acid bacteria (LAB) and pseudomonads. The LAB flora of raw milk was dominated by enterococci (40.8%), followed by lactococci (20.4%), leuconostocs (18.4%), and mesophilic lactobacilli (10.2%). Enterococcus faecalis (30.1%) and Enterococcus faecium (13.7%) were the most common LAB isolates, followed by Enterococcus durans, Lactococcus lactis subsp. lactis, Lactobacillus plantarum, and Leuconostoc lactis. Thermization at 60 degrees C for 30 s was effective for reducing raw milk contamination by enterobacteria (5.1 log CFU/ml), coagulase-positive staphylococci (3.3 log CFU/ml), and Listeria (present in 25-ml samples) to safe levels, but it also reduced mesophilic lactococci, leuconostocs, lactobacilli, and selected enterococci (72.0%) in thermized milk. Thermization at 67 degrees C for 30 s had a major inactivation effect on all bacterial groups. Two nisin-producing L. lactis subsp. lactis strains (M78 and M104) were isolated from raw milk, but neither nisin-producing nor other bacteriocin-producing LAB strains were isolated from thermized milk. Thus, thermization treatments control harmful bacteria but also may have a negative impact on milk quality by reducing desirable LAB and the biodiversity of raw milk bacteria overall, inactivating potentially protective LAB strains and enhancing the ability of potentially pathogenic enterococci to grow in fresh cheese curds.

Efficacy of UV light treatment for the microbiological decontamination of chicken, associated packaging, and contact surfaces.

PubMed

Haughton, P N; Lyng, J G; Cronin, D A; Morgan, D J; Fanning, S; Whyte, P

2011-04-01

UV light was investigated for the decontamination of raw chicken, associated packaging, and contact surfaces. The UV susceptibilities of a number of Campylobacter isolates (seven Campylobacter jejuni isolates and three Campylobacter coli isolates), Escherichia coli ATCC 25922, and Salmonella enterica serovar Enteritidis ATCC 10376 in liquid media were also investigated. From an initial level of 7 log CFU/ml, no viable Campylobacter cells were detected following exposure to the most intense UV dose (0.192 J/cm(2)) in liquid media (skim milk subjected to ultrahigh-temperature treatment and diluted 1:4 with maximum recovery diluent). Maximum reductions of 4.8 and 6.2 log CFU/ml were achieved for E. coli and serovar Enteritidis, respectively, in liquid media. Considerable differences in susceptibilities were found between the Campylobacter isolates examined, with variations of up to 4 log CFU/ml being observed. UV treatment of raw chicken fillet (0.192 J/cm(2)) reduced C. jejuni, E. coli, serovar Enteritidis, total viable counts, and Enterobacteriaceae by 0.76, 0.98, 1.34, 1.76, and 1.29 log CFU/g, respectively. Following UV treatment of packaging and surface materials, reductions of up to 3.97, 4.50, and 4.20 log CFU/cm(2) were obtained for C. jejuni, E. coli, and serovar Enteritidis, respectively (P < 0.05). Overall, the color of UV-treated chicken was not significantly affected (P ≥ 0.05). The findings of this study indicate that Campylobacter is susceptible to UV technology and that differences in sensitivities exist between investigated isolates. Overall, UV could be used for improving the microbiological quality of raw chicken and for decontaminating associated packaging and surface materials.

Inactivation of Mycobacterium paratuberculosis by Pulsed Electric Fields

PubMed Central

Rowan, Neil J.; MacGregor, Scott J.; Anderson, John G.; Cameron, Douglas; Farish, Owen

2001-01-01

The influence of treatment temperature and pulsed electric fields (PEF) on the viability of Mycobacterium paratuberculosis cells suspended in 0.1% (wt/vol) peptone water and in sterilized cow's milk was assessed by direct viable counts and by transmission electron microscopy (TEM). PEF treatment at 50°C (2,500 pulses at 30 kV/cm) reduced the level of viable M. paratuberculosis cells by approximately 5.3 and 5.9 log10 CFU/ml in 0.1% peptone water and in cow's milk, respectively, while PEF treatment of M. paratuberculosis at lower temperatures resulted in less lethality. Heating alone at 50°C for 25 min or at 72°C for 25 s (extended high-temperature, short-time pasteurization) resulted in reductions of M. paratuberculosis of approximately 0.01 and 2.4 log10 CFU/ml, respectively. TEM studies revealed that exposure to PEF treatment resulted in substantial damage at the cellular level to M. paratuberculosis. PMID:11375202

Survival and detection of Bacillus cereus in the presence of Escherichia coli, Salmonella enteritidis, Staphylococcus aureus, Pseudomonas aeruginosa and Candida albicans after rechallenge in make-up removers.

PubMed

Yossa, N; Arce, G; Smiley, J; Jo Huang, M-C; Yin, L; Bell, R; Tallent, S; Brown, E; Hammack, T

2018-02-01

Pathogenic contamination of cosmetics intended to be applied on or around the eye area, including make-up removers, may lead to severe eye infections. To assess the efficacy of antimicrobial preservatives in these products, we investigated the survival and detection of Bacillus cereus F 4227A spiked into make-up removers, alone and in the presence of other relevant micro-organisms. Four brands of make-up removers, A, B, C and D, were challenged three times (day 0, day 7 and day 14) using B. cereus, in pure and mixed cultures, at a final concentration of 5 log CFU per mL of Bacillus cereus or 6 log CFU per mL for other micro-organisms. Inoculated samples were diluted and spiral-plated after 30 min and 24 h of each challenge onto selective media for recovery of surviving micro-organisms: BACARA (B. cereus), MacConkey (E. coli), ChromID (P. aeruginosa), XLT4 (S. enteritidis), Baird Parker agar (Staph. aureus) and PDA+chlortetracycline HCL (C. albicans). The population of B. cereus spiked as a pure culture increased significantly from the first to the third challenge after 30-min exposure time, going from 0.73 to 2.59 in A, from 0.80 to 2.69 in B and from 0.80 to 1.67 log CFU per mL in C (P < 0.05). Likewise, the B. cereus population from the mixed cultures had a significantly higher survival count at the third challenge: from 0.12 log MPN per mL to 2.16 log CFU per mL in A, 0.57 to 2.27 log CFU per mL in B and from undetected (LOD = 0.48 log MPN) to 0.98 log CFU per mL in C, respectively. After challenges, Staph. aureus, C. albicans and P. aeruginosa increased in B; Staph. aureus and C. albicans in C; and E. coli and Staph. aureus in D. The growth of other bacteria types was unaffected by the number of challenges, but B. cereus population was detected with the third challenge. It is appropriate to assess the antimicrobial efficacy of preservatives using at least three challenges, especially for cosmetics that are subjected to repetitive contamination by users. © 2017 U.S. Food and Drug Administration. International Journal of Cosmetic Science published by John Wiley & Sons Ltd on behalf of Society of Cosmetic Scientists and the Société Française de Cosmétologie.

Evaluation of sanitizers for inactivating Salmonella on in-shell pecans and pecan nutmeats.

PubMed

Beuchat, Larry R; Mann, David A; Alali, Walid Q

2012-11-01

Chlorine, organic acids, and water extracts of inedible pecan components were tested for effectiveness in killing Salmonella on pecans. In-shell pecans and nutmeats (U.S. Department of Agriculture medium pieces) were immersion inoculated with a mixture of five Salmonella serotypes, dried to 3.7% moisture, and stored at 4°C for 3 to 6 weeks. In-shell nuts were immersed in chlorinated water (200, 400, and 1,000 μg/ml), lactic acid (0.5, 1, and 2%), and levulinic acid (0.5, 1, and 2%) with and without 0.05% sodium dodecyl sulfate (SDS), and a mixed peroxyacid sanitizer (Tsunami 200, 40 μg/ml) for up to 20 min at 21°C. The rate of reduction of free chlorine in conditioning water decreased as the ratio of in-shell nuts/water was increased. The rate of reduction was more rapid when nuts were not precleaned before treatment. The initial population of Salmonella on in-shell nuts (5.9 to 6.3 log CFU/g) was reduced by 2.8 log CFU/g after treating with chlorinated water (1,000 μg/ml). Treatment with 2% lactic acid plus SDS or 2% levulinic acid plus SDS reduced the pathogen by 3.7 and 3.4 log CFU/g, respectively. Lactic and levulinic acids (2%) without SDS were less effective (3.3- and 2.1-log CFU/g reductions, respectively) than acids with SDS. Treatment with Tsunami 200 resulted in a 2.4-log CFU/g reduction. In-shell nuts and nutmeats were immersed in water extracts of ground pecan shucks (hulls), shells, a mixture of shells and pith, and pith. The general order of lethality of extracts to Salmonella was shuck < shell-pith ≤ shell ≤ pith < chlorine (400 μg/ml) and shuck < shell ≤ pith = shell-pith < chlorine (400 μg/ml). Results emphasize the importance of removing soil and dust on in-shell pecans before conditioning in chlorinated water and the need for sanitizers with increased effectiveness in killing Salmonella on pecans.

Inactivation of Escherichia coli O157:H7 and Listeria monocytogenes in biofilms by pulsed ultraviolet light.

PubMed

Montgomery, Nedra L; Banerjee, Pratik

2015-06-10

The inactivation of biofilms formed by pathogenic bacteria on ready-to-eat and minimally processed fruits and vegetables by nonthermal processing methods is critical to ensure food safety. Pulsed ultraviolet (PUV) light has shown promise in the surface decontamination of liquid, powdered, and solid foods. In this study, the antimicrobial efficacy of PUV light treatment on nascent biofilms formed by Escherichia coli O157:H7 and Listeria monocytogenes on the surfaces of food packaging materials, such as low-density polyethylene (LDPE), and fresh produce, such as lettuce (Lactuca sativa) leaves, was investigated. The formation of biofilms on Romaine lettuce leaves and LDPE films was confirmed by crystal violet and Alcian blue staining methods. Inactivation of cells in the biofilm was determined by standard plating procedures, and by a luminescence-based bacterial cell viability assay. Upon PUV treatment of 10 s at two different light source to sample distances (4.5 and 8.8 cm), viable cell counts of L. monocytogenes and E. coli O157:H7 in biofilms on the lettuce surface were reduced by 0.6-2.2 log CFU mL(-1) and 1.1-3.8 log CFU mL(-1), respectively. On the LDPE surface, the efficiency of inactivation of biofilm-encased cells was slightly higher. The maximum values for microbial reduction on LDPE were 2.7 log CFU mL(-1) and 3.9 log CFU mL(-1) for L. monocytogenes and E. coli O157:H7, respectively. Increasing the duration of PUV light exposure resulted in a significant (P < 0.05) reduction in biofilm formation by both organisms. The results also revealed that PUV treatment was more effective at reducing E. coli biofilms compared with Listeria biofilms. A moderate increase in temperature (~7-15°C) was observed for both test materials. PUV is an effective nonthermal intervention method for surface decontamination of E. coli O157:H7 and L. monocytogenes on fresh produce and packaging materials.

Survival and growth of Enterobacter sakazakii in infant cereal as affected by composition, reconstitution liquid, and storage temperature.

PubMed

Lin, Li-Chun; Beuchat, Larry R

2007-06-01

Invasive infections caused by Enterobacter sakazakii have occurred predominantly in low-birth-weight neonates and infants younger than 2 months of age. However, infections have also occurred in healthy infants up to 8 months of age and in immunocompromised children up to 4 years of age. The ability of E. sakazakii to survive and grow in infant cereals as affected by composition of the cereal, composition of the reconstitution liquid, and temperature is unknown. A study was done to determine the survival and growth characteristics of E. sakazakii initially at populations of 0.005 and 0.52 CFU/ml of infant rice cereal, oatmeal cereal, or rice with mixed fruit cereal reconstituted with water, milk, or apple juice. Reconstituted cereals were stored at 4, 12, 21, and 30 degrees C, and populations were monitored for up to 72 h. Growth did not occur in reconstituted cereals stored at 4 degrees C or in cereals reconstituted with apple juice and stored at 12 degrees C. Populations (> or =1 CFU/ml) were detected in cereals reconstituted with water or milk and stored at 12, 21, and 30 degres C for 24, 8, and 4 h, respectively. The composition of infant cereals did not markedly affect the survival or growth of E. sakazakii in reconstituted cereals. Populations of E. sakazakii in reconstituted cereal decreased with increases in populations of mesophilic aerobic microflora up to 8 to 9 log CFU/ml, which was concurrent with decreases in pH. E. sakazakii, initially at 2.62 log CFU/ml of rice cereal reconstituted with apple juice (pH 4.32), survived at 40C for at least 14 days. The pathogen grew at 21 and 30 degrees C within 2 days and then decreased to undetectable levels (<1 CFU/10 ml) in cereal stored at 21 degrees C for 5 days or 30'C for 4 days. Initially, at 7.32 log CFU/ml, E. sakazakii was detected in rice cereal stored at 4 degrees C for 50 days. It is recommended that reconstituted infant cereals stored at 21 or 30 degrees C be discarded within 4 h after preparation or stored at -40C, temperatures at which E. sakazakii will not grow.

Survival of a native toxigenic isolate of Listeria monocytogenes CFR 1302 during storage of milk-based foods can be a potential cause of health risk.

PubMed

Jayanth, Hampapura S; Varadaraj, Mandyam C

2017-07-01

The ability of a native toxigenic culture of Listeria monocytogenes CFR 1302 to survive and elaborate associated toxigenic trait in ice cream and mango pulp-based lactic fermented milk was studied. The culture of L. monocytogenes inoculated at two initial levels of 4.6 and 5.6 log 10  CFU/ml almost remained unaltered during storage of the food products. However, in both the milk-based products, a marginal increase in viable population was observed during 2-4 d of storage as against the initial inoculum levels. The toxigenic trait, listeriolysin "O" was detected by PCR based on species-specific hlyA primers in the two products without any step of enrichment. The positive amplification in PCR was evidenced with initial population levels of 6.3, 7.3, and 8.3 log 10  CFU/ml of the respective products. In culture broth, PCR detection was positive with the lowest level of 2.3 log 10  CFU/ml. The established pathogenic strain of L. monocytogenes Scott A used as a reference culture revealed almost the same behavior to that of native culture in the food products. The findings of present study bring into focus that, irrespective of low storage temperatures, there exists the potential health hazard associated with foods initially contaminated with risk population levels of L. monocytogenes.

Effect of surface roughness and stainless steel finish on Listeria monocytogenes attachment and biofilm formation.

PubMed

Rodriguez, Andres; Autio, Wesley R; McLandsborough, Lynne A

2008-01-01

The purpose of this study was to evaluate the effect of surface roughness (Ra) and finish of mechanically polished stainless steel (Ra = 0.26 +/- 0.05, 0.49 +/- 0.10, and 0.69 +/- 0.05 microm) and electropolished stainless steel (Ra = 0.16 +/- 0.06, 0.40 +/- 0.003, and 0.67 +/- 0.02 microm) on Listeria adhesion and biofilm formation. A four-strain cocktail of Listeria monocytogenes was used. Each strain (0.1%) was added to 200 ml of tryptic soy broth (TSB), and coupons were inserted to the mixture for 5 min. For biofilm formation, coupons with adhesive cells were incubated in 1:20 diluted TSB at 32 degrees C for 48 h. The experiment was performed by a randomized block design. Our results show that the level of Listeria present after 48 h of incubation (mean = 7 log CFU/cm2) was significantly higher than after 5 min (mean = 6.0 log CFU/cm2) (P < 0.01). No differences in initial adhesion were seen in mechanically finished (mean = 6.7 log CFU/cm2) when compared with electropolished stainless steel (mean = 6.7 log CFU/cm2) (P > 0.05). Listeria initial adhesion (values ranged from 5.9 to 6.1 log CFU/cm2) or biofilm formation (values ranged from 6.9 to 7.2 log CFU/cm2) was not significantly correlated with Ra values (P > 0.05). Image analysis with an atomic force microscope showed that bacteria did not colonize the complete surface after 48 h but were individual cells or grouped in microcolonies that ranged from 5 to 10 microm in diameter and one to three cell layers in thickness. Exopolymeric substances were observed to be associated with the colonies. According to our results, electropolishing stainless steel does not pose a significant advantage for food sanitation over mechanically finished stainless steel.

Impact of spores on the comparative efficacies of five antibiotics for treatment of Bacillus anthracis in an in vitro hollow fiber pharmacodynamic model.

PubMed

Louie, Arnold; VanScoy, Brian D; Brown, David L; Kulawy, Robert W; Heine, Henry S; Drusano, George L

2012-03-01

Bacillus anthracis, the bacterium that causes anthrax, is an agent of bioterrorism. The most effective antimicrobial therapy for B. anthracis infections is unknown. An in vitro pharmacodynamic model of B. anthracis was used to compare the efficacies of simulated clinically prescribed regimens of moxifloxacin, linezolid, and meropenem with the "gold standards," doxycycline and ciprofloxacin. Treatment outcomes for isogenic spore-forming and non-spore-forming strains of B. anthracis were compared. Against spore-forming B. anthracis, ciprofloxacin, moxifloxacin, linezolid, and meropenem reduced the B. anthracis population by 4 log(10) CFU/ml over 10 days. Doxycycline reduced the population of this B. anthracis strain by 5 log(10) CFU/ml (analysis of variance [ANOVA] P = 0.01 versus other drugs). Against an isogenic non-spore-forming strain, meropenem killed the vegetative B. anthracis the fastest, followed by moxifloxacin and ciprofloxacin and then doxycycline. Linezolid offered the lowest bacterial kill rate. Heat shock studies using the spore-producing B. anthracis strain showed that with moxifloxacin, ciprofloxacin, and meropenem therapies the total population was mostly spores, while the population was primarily vegetative bacteria with linezolid and doxycycline therapies. Spores have a profound impact on the rate and extent of killing of B. anthracis. Against spore-forming B. anthracis, the five antibiotics killed the total (spore and vegetative) bacterial population at similar rates (within 1 log(10) CFU/ml of each other). However, bactericidal antibiotics killed vegetative B. anthracis faster than bacteriostatic drugs. Since only vegetative-phase B. anthracis produces the toxins that may kill the infected host, the rate and mechanism of killing of an antibiotic may determine its overall in vivo efficacy. Further studies are needed to examine this important observation.

Impact of Spores on the Comparative Efficacies of Five Antibiotics for Treatment of Bacillus anthracis in an In Vitro Hollow Fiber Pharmacodynamic Model

PubMed Central

VanScoy, Brian D.; Brown, David L.; Kulawy, Robert W.; Heine, Henry S.; Drusano, George L.

2012-01-01

Bacillus anthracis, the bacterium that causes anthrax, is an agent of bioterrorism. The most effective antimicrobial therapy for B. anthracis infections is unknown. An in vitro pharmacodynamic model of B. anthracis was used to compare the efficacies of simulated clinically prescribed regimens of moxifloxacin, linezolid, and meropenem with the “gold standards,” doxycycline and ciprofloxacin. Treatment outcomes for isogenic spore-forming and non-spore-forming strains of B. anthracis were compared. Against spore-forming B. anthracis, ciprofloxacin, moxifloxacin, linezolid, and meropenem reduced the B. anthracis population by 4 log10 CFU/ml over 10 days. Doxycycline reduced the population of this B. anthracis strain by 5 log10 CFU/ml (analysis of variance [ANOVA] P = 0.01 versus other drugs). Against an isogenic non-spore-forming strain, meropenem killed the vegetative B. anthracis the fastest, followed by moxifloxacin and ciprofloxacin and then doxycycline. Linezolid offered the lowest bacterial kill rate. Heat shock studies using the spore-producing B. anthracis strain showed that with moxifloxacin, ciprofloxacin, and meropenem therapies the total population was mostly spores, while the population was primarily vegetative bacteria with linezolid and doxycycline therapies. Spores have a profound impact on the rate and extent of killing of B. anthracis. Against spore-forming B. anthracis, the five antibiotics killed the total (spore and vegetative) bacterial population at similar rates (within 1 log10 CFU/ml of each other). However, bactericidal antibiotics killed vegetative B. anthracis faster than bacteriostatic drugs. Since only vegetative-phase B. anthracis produces the toxins that may kill the infected host, the rate and mechanism of killing of an antibiotic may determine its overall in vivo efficacy. Further studies are needed to examine this important observation. PMID:22155821

Hand hygiene with soap and water is superior to alcohol rub and antiseptic wipes for removal of Clostridium difficile.

PubMed

Oughton, Matthew T; Loo, Vivian G; Dendukuri, Nandini; Fenn, Susan; Libman, Michael D

2009-10-01

To evaluate common hand hygiene methods for efficacy in removing Clostridium difficile. Randomized crossover comparison among 10 volunteers with hands experimentally contaminated by nontoxigenic C. difficile. Interventions included warm water with plain soap, cold water with plain soap, warm water with antibacterial soap, antiseptic hand wipes, alcohol-based handrub, and a control involving no intervention. All interventions were evaluated for mean reduction in colony-forming units (CFUs) under 2 contamination protocols: "whole hand" and "palmar surface." Results were analyzed according to a Bayesian approach, by using hierarchical models adjusted for multiple observations. Under the whole-hand protocol, the greatest adjusted mean reductions were achieved by warm water with plain soap (2.14 log(10) CFU/mL [95% credible interval (CrI), 1.74-2.54 log(10) CFU/mL]), cold water with plain soap (1.88 log(10) CFU/mL [95% CrI, 1.48-2.28 log(10) CFU/mL), and warm water with antibacterial soap (1.51 log(10) CFU/mL [95% CrI, 1.12-1.91 log(10) CFU/mL]), followed by antiseptic hand wipes (0.57 log(10) CFU/mL [95% CrI, 0.17-0.96 log(10) CFU/mL]). Alcohol-based handrub (0.06 log(10) CFU/mL [95% CrI, -0.34 to 0.45 log(10) CFU/mL]) was equivalent to no intervention. Under the palmar surface protocol, warm water with plain soap, cold water with plain soap, and warm water with antibacterial soap again yielded the greatest mean reductions, followed by antiseptic hand wipes (26.6, 26.6, 26.6, and 21.9 CFUs per plate, respectively), when compared with alcohol-based handrub. Hypothenar (odds ratio, 10.98 [95% CrI, 1.96-37.65]) and thenar (odds ratio, 6.99 [95% CrI, 1.25-23.41]) surfaces were more likely than fingertips to remain heavily contaminated after handwashing. Handwashing with soap and water showed the greatest efficacy in removing C. difficile and should be performed preferentially over the use of alcohol-based handrubs when contact with C. difficile is suspected or likely.

Activities of Tigecycline (GAR-936) against Legionella pneumophila In Vitro and in Guinea Pigs with L. pneumophila Pneumonia

PubMed Central

Edelstein, Paul H.; Weiss, William J.; Edelstein, Martha A. C.

2003-01-01

The activities of tigecycline (Wyeth Research) against extracellular and intracellular Legionella pneumophila and for the treatment of guinea pigs with L. pneumophila pneumonia were studied. The tigecycline MIC at which 50% of strains are inhibited for 101 different Legionella sp. strains was 4 μg/ml versus 0.125 and 0.25 μg/ml for azithromycin and erythromycin, respectively. Tigecycline was about as active as erythromycin (tested at 1 μg/ml) against the F889 strain of L. pneumophila grown in guinea pig alveolar macrophages and more active than erythromycin against the F2111 strain. Azithromycin (0.25 μg/ml) was more active than (F889) or as active as (F2111) tigecycline (1 μg/ml) in the macrophage model. When tigecycline was given (7.5 mg/kg of body weight subcutaneously once) to guinea pigs with L. pneumophila pneumonia, the mean peak serum and lung levels were 2.3 and 1.8 μg/ml (1.2 and 1.5 μg/g) at 1 and 2 h postinjection, respectively. The serum and lung areas under the concentration time curve from 0 to 24 h were 13.7 and 15.8 μg · h/ml, respectively. Thirteen of 16 guinea pigs with L. pneumophila pneumonia treated with tigecycline (7.5 mg/kg subcutaneously once daily for 5 days) survived for 7 days post-antimicrobial therapy, as did 11 of 12 guinea pigs treated with azithromycin (15 mg/kg intraperitoneally once daily for 2 days). None of 12 guinea pigs treated with saline survived. Tigecycline-treated guinea pigs had average end of therapy lung counts of 1 × 106 CFU/g (range, 2.5 × 104 to 3.2 × 106 CFU/g) versus <1 × 102 CFU/g for azithromycin (range, undetectable to 100 CFU/g). A second guinea pig study examined the ability of tigecycline to clear L. pneumophila from the lung after 5 to 9 days of therapy; bacterial concentrations 1 day posttherapy ranged from log10 4.2 to log10 5.5 CFU/g for four different dosing regimens. Tigecycline is about as effective as erythromycin against intracellular L. pneumophila, but tigecycline inactivation by the test media confounded the interpretation of susceptibility data. Tigecycline was effective at preventing death from pneumonia in an animal model of Legionnaires' disease, warranting human clinical trials of the drug for the disease. PMID:12543655

Quantifying the Effects of Water Temperature, Soap Volume, Lather Time, and Antimicrobial Soap as Variables in the Removal of Escherichia coli ATCC 11229 from Hands.

PubMed

Jensen, Dane A; Macinga, David R; Shumaker, David J; Bellino, Roberto; Arbogast, James W; Schaffner, Donald W

2017-06-01

The literature on hand washing, while extensive, often contains conflicting data, and key variables are only superficially studied or not studied at all. Some hand washing recommendations are made without scientific support, and agreement between recommendations is limited. The influence of key variables such as soap volume, lather time, water temperature, and product formulation on hand washing efficacy was investigated in the present study. Baseline conditions were 1 mL of a bland (nonantimicrobial) soap, a 5-s lather time, and 38°C (100°F) water temperature. A nonpathogenic strain of Escherichia coli (ATCC 11229) was the challenge microorganism. Twenty volunteers (10 men and 10 women) participated in the study, and each test condition had 20 replicates. An antimicrobial soap formulation (1% chloroxylenol) was not significantly more effective than the bland soap for removing E. coli under a variety of test conditions. Overall, the mean reduction was 1.94 log CFU (range, 1.83 to 2.10 log CFU) with the antimicrobial soap and 2.22 log CFU (range, 1.91 to 2.54 log CFU) with the bland soap. Overall, lather time significantly influenced efficacy in one scenario, in which a 0.5-log greater reduction was observed after 20 s with bland soap compared with the baseline wash (P = 0.020). Water temperature as high as 38°C (100°F) and as low as 15°C (60°F) did not have a significant effect on the reduction of bacteria during hand washing; however, the energy usage differed between these temperatures. No significant differences were observed in mean log reductions experienced by men and women (both 2.08 log CFU; P = 0.988). A large part of the variability in the data was associated with the behaviors of the volunteers. Understanding what behaviors and human factors most influence hand washing may help researchers find techniques to optimize the effectiveness of hand washing.

Titanium dioxide/UV photocatalytic disinfection in fresh carrots.

PubMed

Cho, Mihee; Choi, Yoonjung; Park, Hyojin; Kim, Kwansik; Woo, Gun-Jo; Park, Jiyong

2007-01-01

Increased occurrences of fresh produce-related outbreaks of foodborne illness have focused attention on effective washing processes for fruits and vegetables. A titanium dioxide (TiO2) photocatalytic reaction under UV radiation provides a high rate of disinfection. The photo-killing effects of TiO2 on bacteria in liquid cultures under experimental conditions have been widely studied. However, the disinfection effects of the TiO2 photocatalytic reaction on fresh vegetables during a washing process have not been evaluated. Our objectives were to design a pilot-scale TiO2/UV photocatalytic reactor for fresh carrots and to compare the bactericidal effects of the TiO2/UV reaction against bacteria in liquid media and on carrots. TiO2/UV photocatalytic reactions for 40, 60, and 30 s were required for the complete killing of Escherichia coli, Salmonella Typhimurium, and Bacillus cereus (initial counts of approximately 6.7 log CFU/ml), respectively. The counts of total aerobic bacteria in fresh carrots and foodborne pathogenic bacteria in inoculated carrots were also measured. Counts of total aerobic bacteria were reduced by 1.8 log CFU/g after TiO2/UV photocatalytic disinfection for 20 min compared with a 1.1-log CFU/g reduction by UV alone. E. coli, Salmonella Typhimurium, and B. cereus (8 log CFU/ml) were inoculated onto carrots, and the number of surviving bacteria in carrots was determined after treatment. The TiO2/UV treatment exhibited 2.1-, 2.3-, and 1.8-log CFU/g reductions in the counts of E. coli, Salmonella Typhimurium, and B. cereus, respectively, compared with 1.3-, 1.2-, and 1.2-log CFU/g reductions by UV alone. The TiO2/UV photocatalyst reaction showed significant bactericidal effects, indicating that this process is applicable to nonthermal disinfection of fresh vegetables.

Predicting acute uncomplicated urinary tract infection in women: a systematic review of the diagnostic accuracy of symptoms and signs.

PubMed

Giesen, Leonie G M; Cousins, Gráinne; Dimitrov, Borislav D; van de Laar, Floris A; Fahey, Tom

2010-10-24

Acute urinary tract infections (UTI) are one of the most common bacterial infections among women presenting to primary care. However, there is a lack of consensus regarding the optimal reference standard threshold for diagnosing UTI. The objective of this systematic review is to determine the diagnostic accuracy of symptoms and signs in women presenting with suspected UTI, across three different reference standards (10(2) or 10(3) or 10(5) CFU/ml). We also examine the diagnostic value of individual symptoms and signs combined with dipstick test results in terms of clinical decision making. Searches were performed through PubMed (1966 to April 2010), EMBASE (1973 to April 2010), Cochrane library (1973 to April 2010), Google scholar and reference checking.Studies that assessed the diagnostic accuracy of symptoms and signs of an uncomplicated UTI using a urine culture from a clean-catch or catherised urine specimen as the reference standard, with a reference standard of at least ≥ 10(2) CFU/ml were included. Synthesised data from a high quality systematic review were used regarding dipstick results. Studies were combined using a bivariate random effects model. Sixteen studies incorporating 3,711 patients are included. The weighted prior probability of UTI varies across diagnostic threshold, 65.1% at ≥ 10(2) CFU/ml; 55.4% at ≥ 10(3) CFU/ml and 44.8% at ≥ 10(2) CFU/ml ≥ 10(5) CFU/ml. Six symptoms are identified as useful diagnostic symptoms when a threshold of ≥ 10(2) CFU/ml is the reference standard. Presence of dysuria (+LR 1.30 95% CI 1.20-1.41), frequency (+LR 1.10 95% CI 1.04-1.16), hematuria (+LR 1.72 95%CI 1.30-2.27), nocturia (+LR 1.30 95% CI 1.08-1.56) and urgency (+LR 1.22 95% CI 1.11-1.34) all increase the probability of UTI. The presence of vaginal discharge (+LR 0.65 95% CI 0.51-0.83) decreases the probability of UTI. Presence of hematuria has the highest diagnostic utility, raising the post-test probability of UTI to 75.8% at ≥ 10(2) CFU/ml and 67.4% at ≥ 10(3) CFU/ml. Probability of UTI increases to 93.3% and 90.1% at ≥ 10(2) CFU/ml and ≥ 10(3) CFU/ml respectively when presence of hematuria is combined with a positive dipstick result for nitrites. Subgroup analysis shows improved diagnostic accuracy using lower reference standards ≥ 10(2) CFU/ml and ≥ 10(3) CFU/ml. Individual symptoms and signs have a modest ability to raise the pretest-risk of UTI. Diagnostic accuracy improves considerably when combined with dipstick tests particularly tests for nitrites.

Impact of accessory gene regulator (agr) dysfunction on vancomycin pharmacodynamics among Canadian community and health-care associated methicillin-resistant Staphylococcus aureus

PubMed Central

2011-01-01

Background The accessory gene regulator (agr) is a quorum sensing cluster of genes which control colonization and virulence in Staphylococcus aureus. We evaluated agr function in community- (CA) and healthcare-associated (HA) MRSA, to compare the pharmacodynamics and bactericidal activity of vancomycin against agr functional and dysfunctional HA-MRSA and CA-MRSA. Methods 40 clinical isolates of MRSA from the Canadian Nosocomial Infection Surveillance Program were evaluated for delta-haemolysin production, as a surrogate marker of agr function. Time kill experiments were performed for vancomycin at 0 to 64 times the MIC against an initial inoculum of 106 and 108 cfu/ml of agr functional and dysfunctional CA-MRSA and HA-MRSA and these data were fit to a hill-type pharmacodynamic model. Results 15% isolates were agr dysfunctional, which was higher among HA-MRSA (26.3%) versus CA-MRSA (4.76%). Against a low initial inoculum of 106 cfu/ml of CA-MRSA, vancomycin pharmacodynamics were similar among agr functional and dysfunctional strains. However, against a high initial inoculum of 108 cfu/ml, killing activity was notably attenuated against agr dysfunctional CA-MRSA (USA400) and HA-MRSA (USA100). CA-MRSA displayed a 20.0 fold decrease in the maximal reduction in bacterial counts (Emax) which was 3.71 log10 CFU/ml for agr functional vs. 2.41 log10 CFU/ml for agr dysfunctional MRSA (p = 0.0007). Conclusions Dysfunction in agr was less common among CA-MRSA vs. HA-MRSA. agr dysfunction demonstrated an impact on vancomycin bactericidal activity and pharmacodynamics against a high initial inoculum of CA-MRSA and HA-MRSA, which may have implications for optimal antimicrobial therapy against persistent, difficult to treat MRSA infections. PMID:21599878

Investigation of reduction and tolerance capability of lactic acid bacteria isolated from kimchi against nitrate and nitrite in fermented sausage condition.

PubMed

Paik, Hyun-Dong; Lee, Joo-Yeon

2014-08-01

Lactobacillus brevis KGR3111, Lactobacillus curvatus KGR 2103, Lactobacillus plantarum KGR 5105, and Lactobacillus sakei KGR 4108 isolated from kimchi were investigated for their potential to be used as starter culture for fermented sausages with the capability to reduce and tolerate nitrate/nitrite. The reduction capability of tested strains for nitrate was not dramatic. All tested strains, however, showed the capability to produce nitrite reductase with the reduction amount of 58.46-75.80 mg/l of NO(2)(-). L. brevis and L. plantarum showed nitrate tolerance with the highest number of 8.71 log cfu/ml and 8.81 log cfu/ml, and L. brevis and L. sakei exhibited nitrite tolerance with the highest number of 8.24 log cfu/ml and 8.25 log cfu/ml, respectively. As a result, L. brevis, L. plantarum, and L. sakei isolated from kimchi showed a tolerance against nitrate or nitrite with a good nitrite reduction capability, indicating the satisfaction of one of the selection criteria to be used as starter culture for fermented sausages. Copyright © 2014 Elsevier Ltd. All rights reserved.

Microbial Indicator Profiling of Fresh Produce and Environmental Samples from Farms and Packing Facilities in Northern Mexico.

PubMed

Heredia, Norma; Caballero, Cindy; Cárdenas, Carmen; Molina, Karina; García, Rafael; Solís, Luisa; Burrowes, Vanessa; Bartz, Faith E; de Aceituno, Anna Fabiszewski; Jaykus, Lee-Ann; García, Santos; Leon, Juan

2016-07-01

To compare microbiological indicator and pathogen contamination among different types of fresh produce and environmental samples along the production chain, 636 samples of produce (rinsates from cantaloupe melons, jalapeño peppers, and tomatoes) and environmental samples (rinsates from hands of workers, soil, and water) were collected at four successive steps in the production process (from the field before harvest through the packing facility) on 11 farms in northern Mexico during 2011 and 2012. Samples were assayed for enteric pathogens (Escherichia coli O157:H7, other Shiga toxigenic E. coli, Salmonella, and Listeria monocytogenes) and microbial indicators (coliforms, other E. coli strains, and Enterococcus spp.). Salmonella was the only pathogen detected; it was found in one preharvest jalapeño sample (detection limits: 0.0033 CFU/ml in produce and hand samples, 0.0013 CFU/ml in water, and 0.04 CFU/g in soil). Microbial indicator profiles for produce, worker hands, and soil from jalapeño and tomato farms were similar, but cantaloupe farm samples had higher indicator levels (P < 0.05 for all comparisons) on fruit (6.5, 2.8, and 7.2 log CFU per fruit) and hands (6.6, 3.1, and 7.1 log CFU per hand) for coliforms, E. coli, and Enterococcus, respectively, and lower E. coli levels in soil (<1 CFU/g). In water from tomato farms, E. coli indicators were significantly more prevalent (70 to 89% of samples were positive; P = 0.01 to 0.02), and geometric mean levels were higher (0.3 to 0.6 log CFU/100 ml) than those in cantaloupe farm water (32 to 38% of samples were positive, geometric mean <1 CFU/100 ml). Microbial indicators were present during all production steps, but prevalence and levels were generally highest at the final on-farm production step (the packing facility) (P < 0.03 for significant comparisons). The finding that microbial contamination on produce farms is influenced by produce type and production step can inform the design of effective approaches to mitigate microbial contamination.

Fate of Escherichia coli O157:H7 and Salmonella in soil and lettuce roots as affected by potential home gardening practices.

PubMed

Erickson, Marilyn C; Liao, Jean; Payton, Alison S; Webb, Cathy C; Ma, Li; Zhang, Guodong; Flitcroft, Ian; Doyle, Michael P; Beuchat, Larry R

2013-12-01

The survival and distribution of enteric pathogens in soil and lettuce systems were investigated in response to several practices (soil amendment supplementation and reduced watering) that could be applied by home gardeners. Leaf lettuce was grown in manure compost/top soil (0:5, 1:5 or 2:5 w/w) mixtures. Escherichia coli O157:H7 or Salmonella was applied at a low or high dose (10(3) or 10(6) colony-forming units (CFU) mL(-1) ) to the soil of seedlings and mid-age plants. Supplementation of top soil with compost did not affect pathogen survival in the soil or on root surfaces, suggesting that nutrients were not a limiting factor. Salmonella populations on root surfaces were 0.7-0.8 log CFU g(-1) lower for mid-age plants compared with seedlings. E. coli O157:H7 populations on root surfaces were 0.8 log CFU g(-1) lower for mid-age plants receiving 40 mL of water compared with plants receiving 75 mL of water on alternate days. Preharvest internalization of E. coli O157:H7 and Salmonella into lettuce roots was not observed at any time. Based on the environmental conditions and high pathogen populations in soil used in this study, internalization of Salmonella or E. coli O157:H7 into lettuce roots did not occur under practices that could be encountered by inexperienced home gardeners. © 2013 Society of Chemical Industry.

Survival of Escherichia coli O157:H7 during the manufacture and ripening of Cacioricotta goat cheese.

PubMed

Ioanna, F; Quaglia, N C; Storelli, M M; Castiglia, D; Goffredo, E; Storelli, A; De Rosa, M; Normanno, G; Jambrenghi, A Caputi; Dambrosio, A

2018-04-01

The aim of this study was to assess the growth and survival of Escherichia coli O157:H7 during the manufacturing and ripening of Cacioricotta goat cheese. Goat milk was artificially contaminated with E. coli O157:H7 and the bacterial load was monitored from production up to 90 days of ripening. Goat milk was inoculated with 10 2  cfu ml -1 of E. coli O157:H7 and the bacterial count of the curd at time zero was 2.31 log 10  cfu g -1 . During the first day of ripening, the bacterial load has increased to 5.73 log 10  cfu g -1 to more than 6.20 log 10  cfu g -1 during the first week. The bacterial load remained constant up to 28 days and then slightly decreased until the end of ripening, with values of a w and pH of 0.88 and 5.41 respectively. The results of this study highlighted that E. coli O157:H7 is able to survive the manufacturing process and they suggest that the 90-day period of ripening alone is insufficient to remove E. coli O157:H7 in contaminated Cacioricotta goat cheese. Moreover, these results support the assumption that the presence of a low contamination of milk with E. coli O157:H7 could represent a potential source of infection and a threat to consumers. Copyright © 2017 Elsevier Ltd. All rights reserved.

Inhibitory activity of cheese whey fermented with kefir grains.

PubMed

Londero, A; Quinta, R; Abraham, A G; Sereno, R; De Antoni, G; Garrote, G L

2011-01-01

We investigated the chemical and microbiological compositions of three types of whey to be used for kefir fermentation as well as the inhibitory capacity of their subsequent fermentation products against 100 Salmonella sp. and 100 Escherichia coli pathogenic isolates. All the wheys after fermentation with 10% (wt/vol) kefir grains showed inhibition against all 200 isolates. The content of lactic acid bacteria in fermented whey ranged from 1.04 × 10(7) to 1.17 × 10(7) CFU/ml and the level of yeasts from 2.05 × 10(6) to 4.23 × 10(6) CFU/ml. The main changes in the chemical composition during fermentation were a decrease in lactose content by 41 to 48% along with a corresponding lactic acid production to a final level of 0.84 to 1.20% of the total reaction products. The MIC was a 30% dilution of the fermentation products for most of the isolates, while the MBC varied between 40 and 70%, depending on the isolate. The pathogenic isolates Salmonella enterica serovar Enteritidis 2713 and E. coli 2710 in the fermented whey lost their viability after 2 to 7 h of incubation. When pathogens were deliberately inoculated into whey before fermentation, the CFU were reduced by 2 log cycles for E. coli and 4 log cycles for Salmonella sp. after 24 h of incubation. The inhibition was mainly related to lactic acid production. This work demonstrated the possibility of using kefir grains to ferment an industrial by-product in order to obtain a natural acidic preparation with strong bacterial inhibitory properties that also contains potentially probiotic microorganisms.

Physicochemical Quality and Chemical Safety of Chlorine as a Reconditioning Agent and Wash Water Disinfectant for Fresh-Cut Lettuce Washing

PubMed Central

Van Haute, Sam; Holvoet, Kevin; Uyttendaele, Mieke

2013-01-01

Chlorine was assessed as a reconditioning agent and wash water disinfectant in the fresh-cut produce industry. Artificial fresh-cut lettuce wash water, made from butterhead lettuce, was used for the experiments. In the reconditioning experiments, chlorine was added to artificial wash water inoculated with Escherichia coli O157 (6 log CFU/ml). Regression models were constructed based on the inactivation data and validated in actual wash water from leafy vegetable processing companies. The model that incorporated chlorine dose and chemical oxygen demand (COD) of the wash water accurately predicted inactivation. Listeria monocytogenes was more resistant to chlorine reconditioning in artificial wash water than Salmonella spp. and Escherichia coli O157. During the washing process with inoculated lettuce (4 log CFU/g), in the absence of chlorine, there was a rapid microbial buildup in the water that accumulated to 5.4 ± 0.4 log CFU/100 ml after 1 h. When maintaining a residual concentration of 1 mg/liter free chlorine, wash water contamination was maintained below 2.7, 2.5, and 2.5 log CFU/100 ml for tap water and artificial process water with COD values of 500 and 1,000 mg O2/liter, respectively. A model was developed to predict water contamination during the dynamic washing process. Only minor amounts of total trihalomethanes were formed in the water during reconditioning. Total trihalomethanes accumulated to larger amounts in the water during the wash water disinfection experiments and reached 124.5 ± 13.4 μg/liter after 1 h of execution of the washing process in water with a COD of 1,000 mg O2/liter. However, no total trihalomethanes were found on the fresh-cut lettuce after rinsing. PMID:23396332

Physicochemical quality and chemical safety of chlorine as a reconditioning agent and wash water disinfectant for fresh-cut lettuce washing.

PubMed

Van Haute, Sam; Sampers, Imca; Holvoet, Kevin; Uyttendaele, Mieke

2013-05-01

Chlorine was assessed as a reconditioning agent and wash water disinfectant in the fresh-cut produce industry. Artificial fresh-cut lettuce wash water, made from butterhead lettuce, was used for the experiments. In the reconditioning experiments, chlorine was added to artificial wash water inoculated with Escherichia coli O157 (6 log CFU/ml). Regression models were constructed based on the inactivation data and validated in actual wash water from leafy vegetable processing companies. The model that incorporated chlorine dose and chemical oxygen demand (COD) of the wash water accurately predicted inactivation. Listeria monocytogenes was more resistant to chlorine reconditioning in artificial wash water than Salmonella spp. and Escherichia coli O157. During the washing process with inoculated lettuce (4 log CFU/g), in the absence of chlorine, there was a rapid microbial buildup in the water that accumulated to 5.4 ± 0.4 log CFU/100 ml after 1 h. When maintaining a residual concentration of 1 mg/liter free chlorine, wash water contamination was maintained below 2.7, 2.5, and 2.5 log CFU/100 ml for tap water and artificial process water with COD values of 500 and 1,000 mg O2/liter, respectively. A model was developed to predict water contamination during the dynamic washing process. Only minor amounts of total trihalomethanes were formed in the water during reconditioning. Total trihalomethanes accumulated to larger amounts in the water during the wash water disinfection experiments and reached 124.5 ± 13.4 μg/liter after 1 h of execution of the washing process in water with a COD of 1,000 mg O2/liter. However, no total trihalomethanes were found on the fresh-cut lettuce after rinsing.

Pharmacodynamics of 750 mg and 500 mg doses of levofloxacin against ciprofloxacin-resistant strains of Streptococcus pneumoniae.

PubMed

Lister, Philip D

2002-09-01

An in vitro pharmacokinetic model (IVPM) was used to evaluate the pharmacodynamics of the 750 mg and 500 mg doses of levofloxacin against 4 ciprofloxacin-nonsusceptible Streptococcus pneumoniae. Levofloxacin MICs ranged from 1.4 to 3.2 micro g/ml. Log-phase cultures (5 x 10(7) cfu/ml) were inoculated into the IVPM and exposed to the peak free-drug concentrations of levofloxacin achieved in human serum with each dose. Levofloxacin was dosed at 0 and 24 h, elimination pharmacokinetics were simulated, and viable counts were measured over 30 h. The 750 mg dose was rapidly bactericidal against all 4 strains, achieving eradication within 30 h. Against strains with levofloxacin MICs of 1.4 and 1.8 micro g/ml, the 500 mg dose exhibited pharmacodynamics similar to the 750 mg dose. In contrast, against strains with levofloxacin MICs of 2.6 and 3.2 micro g/ml, viable counts never fell below 10(4) cfu/ml. The rapid killing and eradication of these pneumococci by the 750 mg dose warrant the clinical evaluation of this new dose in the treatment of pneumococcal infections.

Antibacterial effect of lactoferricin B on Escherichia coli O157:H7 in ground beef.

PubMed

Venkitanarayanan, K S; Zhao, T; Doyle, M P

1999-07-01

The antibacterial activity of lactoferricin B on enterohemorrhagic Escherichia coli O157:H7 in 1% peptone medium and ground beef was studied at 4 and 10 degrees C. In 1% peptone medium, 50 and 100 microg of lactoferricin B per ml reduced E. coli O157:H7 populations by approximately 0.7 and 2.0 log CFU/ml, respectively. Studies comparing the antibacterial effect of lactoferricin B on E. coli O157:H7 in 1% peptone at pH 5.5 and 7.2 did not reveal any significant difference (P > 0.5) at the two pH values. Lactoferricin B (100 microg/g) reduced E. coli O157:H7 population in ground beef by about 0.8 log CFU/g (P < 0.05). No significant difference (P > 0.5) was observed in the total plate count between treatment and control ground beef samples stored at 4 and 10 degrees C. The antibacterial effect of lactoferricin B on E. coli O157:H7 observed in this study is not of sufficient magnitude to merit its use in ground beef for controlling the pathogen.

Activities of fosfomycin, tigecycline, colistin, and gentamicin against extended-spectrum-β-lactamase-producing Escherichia coli in a foreign-body infection model.

PubMed

Corvec, Stéphane; Furustrand Tafin, Ulrika; Betrisey, Bertrand; Borens, Olivier; Trampuz, Andrej

2013-03-01

Limited antimicrobial agents are available for the treatment of implant-associated infections caused by fluoroquinolone-resistant Gram-negative bacilli. We compared the activities of fosfomycin, tigecycline, colistin, and gentamicin (alone and in combination) against a CTX-M15-producing strain of Escherichia coli (Bj HDE-1) in vitro and in a foreign-body infection model. The MIC and the minimal bactericidal concentration in logarithmic phase (MBC(log)) and stationary phase (MBC(stat)) were 0.12, 0.12, and 8 μg/ml for fosfomycin, 0.25, 32, and 32 μg/ml for tigecycline, 0.25, 0.5, and 2 μg/ml for colistin, and 2, 8, and 16 μg/ml for gentamicin, respectively. In time-kill studies, colistin showed concentration-dependent activity, but regrowth occurred after 24 h. Fosfomycin demonstrated rapid bactericidal activity at the MIC, and no regrowth occurred. Synergistic activity between fosfomycin and colistin in vitro was observed, with no detectable bacterial counts after 6 h. In animal studies, fosfomycin reduced planktonic counts by 4 log(10) CFU/ml, whereas in combination with colistin, tigecycline, or gentamicin, it reduced counts by >6 log(10) CFU/ml. Fosfomycin was the only single agent which was able to eradicate E. coli biofilms (cure rate, 17% of implanted, infected cages). In combination, colistin plus tigecycline (50%) and fosfomycin plus gentamicin (42%) cured significantly more infected cages than colistin plus gentamicin (33%) or fosfomycin plus tigecycline (25%) (P < 0.05). The combination of fosfomycin plus colistin showed the highest cure rate (67%), which was significantly better than that of fosfomycin alone (P < 0.05). In conclusion, the combination of fosfomycin plus colistin is a promising treatment option for implant-associated infections caused by fluoroquinolone-resistant Gram-negative bacilli.

Quinolone Resistance Reversion by Targeting the SOS Response

PubMed Central

Recacha, E.; Machuca, J.; Díaz de Alba, P.; Ramos-Güelfo, M.; Docobo-Pérez, F.; Pascual, A.

2017-01-01

ABSTRACT Suppression of the SOS response has been postulated as a therapeutic strategy for potentiating antimicrobial agents. We aimed to evaluate the impact of its suppression on reversing resistance using a model of isogenic strains of Escherichia coli representing multiple levels of quinolone resistance. E. coli mutants exhibiting a spectrum of SOS activity were constructed from isogenic strains carrying quinolone resistance mechanisms with susceptible and resistant phenotypes. Changes in susceptibility were evaluated by static (MICs) and dynamic (killing curves or flow cytometry) methodologies. A peritoneal sepsis murine model was used to evaluate in vivo impact. Suppression of the SOS response was capable of resensitizing mutant strains with genes encoding three or four different resistance mechanisms (up to 15-fold reductions in MICs). Killing curve assays showed a clear disadvantage for survival (Δlog10 CFU per milliliter [CFU/ml] of 8 log units after 24 h), and the in vivo efficacy of ciprofloxacin was significantly enhanced (Δlog10 CFU/g of 1.76 log units) in resistant strains with a suppressed SOS response. This effect was evident even after short periods (60 min) of exposure. Suppression of the SOS response reverses antimicrobial resistance across a range of E. coli phenotypes from reduced susceptibility to highly resistant, playing a significant role in increasing the in vivo efficacy. PMID:29018116

Rapid Detection of Escherichia coli O157:H7 in Fresh Lettuce Based on Localized Surface Plasmon Resonance Combined with Immunomagnetic Separation.

PubMed

Lee, Nari; Choi, Sung-Wook; Chang, Hyun-Joo; Chun, Hyang Sook

2018-05-01

This study presents a method for rapid detection of Escherichia coli O157:H7 in fresh lettuce based on the properties of target separation and localized surface plasmon resonance of immunomagnetic nanoparticles. The multifunctional immunomagnetic nanoparticles enabling simultaneous separation and detection were prepared by synthesizing magnetic nanoparticles (ca. 10 nm in diameter) composed of an iron oxide (Fe 3 O 4 ) core and gold shell and then conjugating these nanoparticles with the anti- E. coli O157:H7 antibodies. The application of multifunctional immunomagnetic nanoparticles for detecting E. coli O157:H7 in a lettuce matrix allowed detection of the presence of <1 log CFU mL -1 without prior enrichment. In contrast, the detection limit of the conventional plating method was 2.74 log CFU mL -1 . The method, which requires no preenrichment, provides an alternative to conventional microbiological detection methods and can be used as a rapid screening tool for a large number of food samples.

Quantitative contamination and transfer of Escherichia coli from foods by houseflies, Musca domestica L. (Diptera: Muscidae).

PubMed

De Jesús, Antonio J; Olsen, Alan R; Bryce, John R; Whiting, Richard C

2004-06-01

The housefly, Musca domestica L. (Diptera: Muscidae), is recognized as an important factor in the dissemination of various infectious diseases such as cholera, shigellosis, and salmonellosis. They can also serve as a cross-contamination vector for other foodborne pathogens. However, the potential for bacterial transfer by houseflies has been demonstrated in a qualitative rather than quantitative manner. In this study, the numbers of bacteria a housefly can carry on its body and transfer to a clean surface after exposure to a sugar-milk aqueous solution, steak, and potato salad contaminated with a fluorescent gene Escherichia coli (8 log10 CFU/ml) were determined. In the first series of experiments to quantify bacterial numbers on the flies, about 40-60 flies were transferred into a sterile cage, exposed to the food for 30 min, the flies immobilized and the attached E. coli on each fly enumerated. Detectable E. coli (>1.7 log10 CFU/fly) were found on 43% (29/67), 53% (23/43), and 62% (32/52) of the flies in the cages with sugar/milk, steak, and potato salad, respectively. For the positive flies, the geometric mean carriage (log10 CFU/fly) was 2.93+/-1.24 for sugar-milk, 3.77+/-1.28 for steak, and 2.25+/-0.64 for the potato salad. In the second series of experiments, the transfer of bacteria by individual flies from contaminated food to the inner surface of a sterile jar per each landing was determined. E. coli transferred from the sugar-milk was 3.5+/-0.7 log10 CFU/fly-landing, 3.9+/-0.7 for steak and 2.61+/-1.16 for the potato salad. From the initial contamination levels of bacteria and the number of transferred bacteria, it can be calculated that flies contaminate clean surfaces with approximately 0.1 mg of food per landing. Copyright 2004 Elsevier B.V.

Comparison of effects of Wasabia japonica and allyl isothiocyanate on the growth of four strains of Vibrio parahaemolyticus in lean and fatty tuna meat suspensions.

PubMed

Hasegawa, N; Matsumoto, Y; Hoshino, A; Iwashita, K

1999-08-01

Lean tuna meat suspensions (LEAN), with a fat content of 0.006%, and fatty tuna meat suspension (FATTY), with a fat content of 3.0% were inoculated with four strains of Vibrio parahaemolyticus and wasabi (Wasabia japonica Matsumura) or allyl isothiocyanate (AIT) was added before incubation at 37 degrees C. During the incubation, viable Vibrio counts were determined on TCBS agar plates. Both LEAN and FATTY suspensions were inoculated with V. parahaemolyticus AOTO-81, (1.28+/-0.20) x 10(2) CFU/ml, followed by addition of 20 mg wasabi/ml, and incubation for 8 h. The viable Vibrio counts were (7.76+/-5.93) x 10(5) CFU/ml in LEAN and (3.50+/-2.65) x 10(1) CFU/ml in FATTY. When the same strain, at (1.18+/-0.22) x 10(2) CFU/ml, was incubated for 8 h with 50.9 microg AIT/ml, viable Vibrio counts were (4.79+/-1.78) x 10(4) CFU/ml in LEAN and (1.80+/-1.30) x 10(1) CFU/ml in FATTY. Growth of the other three strains with wasabi or AIT was shown to be less in FATTY than in LEAN. These results indicate that growth of V. parahaemolyticus is inhibited more in FATTY than in LEAN by wasabi and allyl isothiocyanate.

Association of nuisance filamentous algae Cladophora spp. with E. coli and Salmonella in public beach waters: impacts of UV protection on bacterial survival.

PubMed

Beckinghausen, Aubrey; Martinez, Alexia; Blersch, David; Haznedaroglu, Berat Z

2014-05-01

This study investigated whether filamentous algal species commonly found in nearshore public beach water systems provide protection from natural UV to bacteria present in the same environmental settings. To test this hypothesis, Cladophora spp., a filamentous nuisance algae group causing undesired water quality in the Great Lakes region was selected and its interactions with a non-pathogenic indicator organism Escherichia coli and a pathogenic strain of Salmonella enterica serovar Typhimurium were tested. In laboratory microcosms where the lake environment and natural sunlight conditions were simulated, a 7-log removal of E. coli was observed in only six hours of exposure to UV with an initial seed concentration of 10(3) CFU mL(-1). With the presence of algae, the same log removal was achieved in 16 hours. At higher seed concentrations of 10(5) CFU mL(-1), E. coli survived for two days with an extended survival up to 11 days in the presence of Cladophora spp. S. typhimurium has shown more resilient survival profiles, with the same log removals achieved in 14 and 20 days for low and high seed concentrations respectively, in the absence of algae. Cladophora spp. caused extended protection for S. typhimurium with much less log reductions reported. Algae-mediated protection from UV irradiation was attributed to certain organic carbon exuded from Cladophora spp. In addition, confocal microscopy images confirmed close interaction between bacteria and algae, more prominent with thin filamentous Cladophora spp.

The Activity of Antimicrobial Surfaces Varies by Testing Protocol Utilized

PubMed Central

Campos, Matias D.; Zucchi, Paola C.; Phung, Ann; Leonard, Steven N.; Hirsch, Elizabeth B.

2016-01-01

Background Contaminated hospital surfaces are an important source of nosocomial infections. A major obstacle in marketing antimicrobial surfaces is a lack of efficacy data based on standardized testing protocols. Aim We compared the efficacy of multiple testing protocols against several “antimicrobial” film surfaces. Methods Four clinical isolates were used: one Escherichia coli, one Klebsiella pneumoniae, and two Staphylococcus aureus strains. Two industry methods (modified ISO 22196 and ASTM E2149), a “dried droplet”, and a “transfer” method were tested against two commercially available antimicrobial films, one film in development, an untreated control, and a positive (silver) control film. At 2 (only ISO) and 24 hours following inoculation, bacteria were collected from film surfaces and enumerated. Results Compared to untreated films in all protocols, there were no significant differences in recovery on either commercial brand at 2 or 24 hours after inoculation. The silver surface demonstrated significant microbicidal activity (mean loss 4.9 Log10 CFU/ml) in all methods and time points with the exception of 2 hours in the ISO protocol and the transfer method. Using our novel droplet method, no differences between placebo and active surfaces were detected. The surface in development demonstrated variable activity depending on method, organism, and time point. The ISO demonstrated minimal activity at 2 hours but significant activity at 24 hours (mean 4.5 Log10 CFU/ml difference versus placebo). The ASTEM protocol exhibited significant differences in recovery of staphylococci (mean 5 Log10 CFU/ml) but not Gram-negative isolates (10 fold decrease). Minimal activity was observed with this film in the transfer method. Conclusions Varying results between protocols suggested that efficacy of antimicrobial surfaces cannot be easily and reproducibly compared. Clinical use should be considered and further development of representative methods is needed. PMID:27494336

Cold plasma inactivates Salmonella Stanley and Escherichia coli O157:H7 inoculated on golden delicious apples.

PubMed

Niemira, Brendan A; Sites, Joseph

2008-07-01

Cold plasma generated in a gliding arc was applied to outbreak strains of Escherichia coli O157:H7 and Salmonella Stanley on agar plates and inoculated onto the surfaces of Golden Delicious apples. This novel sanitizing technology inactivated both pathogens on agar plates, with higher flow rate (40 liters/min) observed to be more efficacious than were lower flow rates (20 liters/min), irrespective of treatment time (1 or 2 min). Golden Delicious apples were treated with various flow rates (10, 20, 30, or 40 liters/min) of cold plasma for various times (1, 2, or 3 min), applied to dried spot inoculations. All treatments resulted in significant (P < 0.05) reductions from the untreated control, with 40 liters/min more effective than were lower flow rates. Inactivation of Salmonella Stanley followed a time-dependent reduction for all flow rates. Reductions after 3 min ranged from 2.9 to 3.7 log CFU/ml, close to the limit of detection. For E. coli O157:H7, 40 liters/min gave similar reductions for all treatment times, 3.4 to 3.6 log CFU/ml. At lower flow rates, inactivation was related to exposure time, with 3 min resulting in reductions of 2.6 to 3 log CFU/ml. Temperature increase of the treated apples was related to exposure time for all flow rates. The maximum temperature of any plasma-treated apple was 50.8 degrees C (28 degrees C above ambient), after 20 liters/min for 3 min, indicating that antimicrobial effects were not the result of heat. These results indicate that cold plasma is a nonthermal process that can effectively reduce human pathogens inoculated onto fresh produce.

Microbiological evaluation of ready-to-eat iceberg lettuce during shelf-life and effectiveness of household washing methods

PubMed Central

Bencardino, Daniela; Vitali, Luca Agostino; Petrelli, Dezemona

2018-01-01

The aim of this study was to assess the microbiological quality of ready-to-eat (RTE) iceberg lettuce. Our investigation was based on the consumption tendency of university students considered a target market for this product. A total of 78 RTE samples were collected from chain supermarkets and analysed for the enumeration of aerobic mesophilic count (AMC), Escherichia coli and the detection of Salmonella spp. and Listeria monocytogenes. All samples were negative for the presence of pathogens. The mean value of AMC at the beginning, in the middle and after the expiration date was: 6.88, 8.51 and 8.72 log CFU g-1, respectively. The same investigation was performed on 12 samples of fresh iceberg lettuce samples. No pathogens were found and the mean value of AMC was lower than the RTE category (5.73 log CFU g-1; P<0.05). The effectiveness of 5 washing methods was determined on 15 samples of both fresh and RTE iceberg lettuce. Samples were washed for 15’ and 30’ in tap water (500 mL), tap water with NaCl (4 g/500 mL), tap water with bicarbonate (8 g/500 mL), tap water with vinegar (10 mL/500 mL) and tap water with chlorine-based disinfectant (10 mL/500 mL). A significant bacterial load reduction was recorded for vinegar and disinfectant after 30’ and 15’, respectively. Overall, these results showed that RTE iceberg lettuce is more contaminated than the fresh product. Also, the consumption in the first few days of packaging and after washing with disinfectants reduces the risk for health consumers. PMID:29732325

Effects of management practices on yield and quality of milk from smallholder dairy units in urban and peri-urban Morogoro, Tanzania.

PubMed

Gillah, Kejeri A; Kifaro, George C; Madsen, Jorgen

2014-10-01

A longitudinal study design was used to assess the management, chemical composition of cows' milk and quantify the microbial load of raw milk produced at farm level. Data were collected between December 2010 and September 2011 in Morogoro municipality. Milk samples were collected once every month and analysed for butter fat (BF), crude protein (CP), total solids (TS) and solids non-fat (SNF). Total bacterial count (TBC) and coliform counts (CC) were normalized by log transformation. The average milk yield was 7.0 l/day and was not influenced by feeding systems and breeds. Dairy cows owned by people who had no regular income produced more milk than government employees and retired officers. Means of BF, TS, SNF and CP were similar in different feeding systems. Wet season had significantly higher TBC (5.9 log10 cfu/ml) and CC (2.4 log10 cfu/ml) but feeding systems had no effect. Stocking density influenced TBC but not CC. It can be concluded that dairy cows produced low milk yield and its quality was poor.

Inactivation of viruses and bacteria on strawberries using a levulinic acid plus sodium dodecyl sulfate based sanitizer, taking sensorial and chemical food safety aspects into account.

PubMed

Zhou, Zijin; Zuber, Sophie; Cantergiani, Frédérique; Butot, Sophie; Li, Dan; Stroheker, Thomas; Devlieghere, Frank; Lima, Anthony; Piantini, Umberto; Uyttendaele, Mieke

2017-09-18

The efficacy of levulinic acid (LVA) in combination with sodium dodecyl sulfate (SDS) in removal of foodborne viruses, enteric bacterial pathogens and their surrogates on fresh strawberries was investigated. Inoculated strawberries were treated with potable water, sodium hypochlorite solution (50ppm), 0.5% LVA plus 0.5% SDS solution, and 5% LVA plus 2% SDS solution respectively for 2min, followed by spray-rinsing with potable water. Water washing removed at least 1.0-log of the tested viral and bacterial strains from the strawberries' surfaces. The 50ppm chlorine wash induced 3.4, 1.5 and 2.1-log reductions for hepatitis A virus (HAV), murine norovirus-1 (MNV-1) and MS2 bacteriophage, respectively. In comparison, the tested bacterial strains showed uniform reductions around 1.6-log CFU/ml. The 0.5% LVA plus 0.5% SDS wash induced 2.7, 1.4 and 2.4-log reductions for HAV, MNV-1 and MS2, which were comparable with the reductions induced by chlorine (P>0.05). For bacteria, over 2.0-log reductions were obtained for Enterococcus faecium, Listeria monocytogenes and Salmonella, while Escherichia coli O157:H7 and Escherichia coli P1 showed reductions of 1.9 and 1.8-log CFU/ml. Higher concentration of LVA plus SDS showed no significantly higher reductions (P>0.05). Sensory tests of washed strawberries and chemical residue analysis of LVA on strawberries after washing were also performed. In conclusion, this study demonstrates good performance of 0.5% LVA plus 0.5% SDS to reduce the levels of enteric pathogens if present on strawberries without altering taste and introducing chemical safety issues. Copyright © 2017 Elsevier B.V. All rights reserved.

Efficacy of electrolyzed oxidizing water in inactivating Salmonella on alfalfa seeds and sprouts.

PubMed

Kim, Chyer; Hung, Yen-Con; Brackett, Robert E; Lin, Chyi-Shen

2003-02-01

Studies have demonstrated that electrolyzed oxidizing (EO) water is effective in reducing foodborne pathogens on fresh produce. This study was undertaken to determine the efficacy of EO water and two different forms of chlorinated water (chlorine water from Cl2 and Ca(OCl)2 as sources of chlorine) in inactivating Salmonella on alfalfa seeds and sprouts. Tengram sets of alfalfa seeds inoculated with a five-strain cocktail of Salmonella (6.3 x 10(4) CFU/g) were subjected to 90 ml of deionized water (control), EO water (84 mg/liter of active chlorine), chlorine water (84 mg/liter of active chlorine), and Ca(OCl)2 solutions at 90 and 20,000 mg/liter of active chlorine for 10 min at 24 +/- 2 degrees C. The application of EO water, chlorinated water, and 90 mg/liter of Ca(OCl)2 to alfalfa seeds for 10 min reduced initial populations of Salmonella by at least 1.5 log10 CFU/g. For seed sprouting, alfalfa seeds were soaked in the different treatment solutions described above for 3 h. Ca(OCl)2 (20,000 mg/liter of active chlorine) was the most effective treatment in reducing the populations of Salmonella and non-Salmonella microflora (4.6 and 7.0 log10 CFU/g, respectively). However, the use of high concentrations of chlorine generates worker safety concerns. Also, the Ca(OCl)2 treatment significantly reduced seed germination rates (70% versus 90 to 96%). For alfalfa sprouts, higher bacterial populations were recovered from treated sprouts containing seed coats than from sprouts with seed coats removed. The effectiveness of EO water improved when soaking treatments were applied to sprouts in conjunction with sonication and seed coat removal. The combined treatment achieved 2.3- and 1.5-log10 CFU/g greater reductions than EO water alone in populations of Salmonella and non-Salmonella microflora, respectively. This combination treatment resulted in a 3.3-log10 CFU/g greater reduction in Salmonella populations than the control (deionized water) treatment.

Linezolid Compared with Eperezolid, Vancomycin, and Gentamicin in an In Vitro Model of Antimicrobial Lock Therapy for Staphylococcus epidermidis Central Venous Catheter-Related Biofilm Infections

PubMed Central

Curtin, John; Cormican, Martin; Fleming, Gerard; Keelehan, John; Colleran, Emer

2003-01-01

Central venous catheter (CVC)-related infection (CVC-RI) is a common complication of CVC use. The most common etiological agents of CVC-RI are gram-positive organisms, in particular, staphylococci. An in vitro model for the formation of biofilms by Staphylococcus epidermidis ATCC 35984 on polyurethane coupons in a modified Robbins device was established. Biofilm formation was confirmed by electron microscopy and was quantified by determination of viable counts. Mueller-Hinton broth was replaced with sterile physiological saline (control) or a solution of vancomycin (10 mg/ml), gentamicin (10 mg/ml), linezolid (2 mg/ml), or eperezolid (4 mg/ml). Viable counts were performed with the coupons after exposure to antimicrobials for periods of 24, 72, 168, and 240 h. The mean viable count per coupon following establishment of the biofilm was 4.6 × 108 CFU/coupon, and that after 14 days of exposure to physiological saline was 2.5 × 107 CFU/coupon. On exposure to vancomycin (10 mg/ml), the mean counts were 2.5 × 107 CFU/coupon at 24 h, 4.3 × 106 CFU/coupon at 72 h, 1.4 × 105 CFU/coupon at 168 h, and undetectable at 240 h. With gentamicin (10 mg/ml) the mean counts were 2.7 × 107 CFU/coupon at 24 h, 3.7 × 106 CFU/coupon at 72 h, 8.4 × 106 CFU/coupon at 168 h, and 6.5 × 106 CFU/coupon at 240 h. With linezolid at 2 mg/ml the mean counts were 7.1 × 105 CFU/coupon at 24 h and not detectable at 72, 168, and 240 h. With eperezolid (4 mg/ml) no viable cells were recovered after 168 h. These data suggest that linezolid (2 mg/ml) and eperezolid (4 mg/ml) achieve eradication of S. epidermidis biofilms more rapidly than vancomycin (10 mg/ml) and gentamicin (10 mg/ml). PMID:14506022

Inactivation of Salmonella on pecan nutmeats by hot air treatment and oil roasting.

PubMed

Beuchat, Larry R; Mann, David A

2011-09-01

Studies were done to determine the effectiveness of hot air drying, dry roasting, and oil roasting in killing Salmonella on pecan nutmeats. Pecan halves and pieces were inoculated by immersion in a five-serotype suspension of Salmonella or by surface application of powdered chalk containing the pathogen. Hot air treatment of low-moisture (2.8 to 4.1%) and high-moisture (10.5 to 11.2%) immersion-inoculated nutmeats (initial population, 6.18 to 7.16 log CFU/g) at 120°C for 20 min reduced the number of Salmonella by 1.18 to 1.26 and 1.89 to 2.04 log CFU/g, respectively. However, regardless of the moisture content, hot air treatment of pecan halves containing 0.77 log CFU/g at 120°C for 20 min failed to eliminate Salmonella. Reductions were >7 log CFU/g when dry pieces were dry roasted at 160°C for 15 min. Treatment of halves at 140°C for 20 min, 150°C for 15 min, or 170°C for 10 min reduced Salmonella by 5 log CFU/g. The pathogen was slightly more heat resistant in immersion-inoculated nutmeats than on surface-inoculated nutmeats. Exposure of immersion-inoculated pieces to peanut oil at 127°C for 1.5 min or 132°C for 1.0 min reduced the number of Salmonella by 5 log CFU/g. Treatment of halves at 138°C for 2.0 min reduced Salmonella by 5 log CFU/g; treatment at 132°C for 2.5 to 4.0 min did not always achieve this reduction. Hot air treatment cannot be relied upon to reduce Salmonella by 5 log CFU/g of raw pecan nutmeats without changing sensory qualities. Treatment temperatures and times typically used to oil roast nutmeats appear to be sufficient to reduce Salmonella by 5 log CFU/g.

Activity of Electrical Current in Experimental Propionibacterium acnes Foreign-Body Osteomyelitis.

PubMed

Schmidt-Malan, Suzannah M; Brinkman, Cassandra L; Greenwood-Quaintance, Kerryl E; Karau, Melissa J; Mandrekar, Jayawant N; Patel, Robin

2017-02-01

Foreign-body-associated infections are often difficult to treat, given that the associated microorganisms are in a biofilm state. Previously, we showed that a low-amperage direct electrical current (DC) reduces Propionibacterium acnes biofilms formed on implant-associated materials in vitro In this study, low-amperage DC was compared to ceftriaxone treatment or no treatment in a novel rat femur model of foreign-body osteomyelitis. A platinum implant seeded with a P. acnes biofilm (10 7 CFU/cm 2 ) and 10 9 CFU of planktonic P. acnes was placed in the femoral medullary cavity. One week later, rats were assigned to one of three treatment groups: no treatment, ceftriaxone treatment, or 200-μA-DC treatment. After 2 weeks of treatment, there were fewer bacteria in the bones of the ceftriaxone group (3.06 log 10 CFU/g of bone [P = 0.0209]) and the 200-μA-DC group (0.5 log 10 CFU/g [P = 0.0015]) than in those of the control group (6.58 log 10 CFU/g). The DC-exposed animals exhibited fewer bacteria than the ceftriaxone-treated animals (P = 0.0330). There were fewer bacteria on the implanted wires in the groups treated with ceftriaxone (0.1 log 10 CFU/cm 2 ) or a 200-μA DC (0.1 log 10 CFU/cm 2 ) than in the control group (2.53 log 10 CFU/cm 2 [P, 0.0003 for both comparisons]). Low-amperage DC may be useful for treating, or aiding in the treatment of, foreign-body infections caused by P. acnes. Copyright © 2017 American Society for Microbiology.

Evaluation of consumable household products for decontaminating retail skinless, boneless chicken breasts.

PubMed

McKee, L H; Neish, L; Pottenger, A; Flores, N; Weinbrenner, K; Remmenga, M

2005-03-01

The effect of 15 consumable products, including juices, wines, and vinegar, used as rinsing agents on microbial loads of retail skinless, boneless chicken breasts was evaluated in two studies. Ten breasts were rinsed for 1 min with each solution. Samples were swabbed before and after rinsing with a cellulose sponge and evaluated for total aerobic (APC), total coliform (TCC), and generic Escherichia coli counts by Petrifilm methods. No differences were found in initial APC or TCC in either study, with initial mean APC ranging from 5.30 to 7.05 log CFU/cm2 and initial mean TCC ranging from 2.21 to 3.36 log CFU/cm2. In study 1, the APC for breasts rinsed with distilled white vinegar (3.22 log CFU/cm2) was lower than for those rinsed with all other solutions except cranberry juice cocktail (3.86 log CFU/cm2). The TCC for breasts rinsed with distilled white vinegar (0.00 log CFU/cm2) and cranberry juice cocktail (0.20 log CFU/cm2) were lower than those for all other solutions except 10% NaCl (0.43 log CFU/cm2) and 10% NaHCO3 (0.48 log CFU/cm2). In study 2, APC values for breasts rinsed with red wine (5.29 log CFU/cm2) and white wine (5.32 log CFU/cm2) were lower than for breasts rinsed with the other three solutions. The TCC after rinsing with chicken broth (4.48 log CFU/cm2) was higher than for all other solutions except Italian dressing. Although distilled white vinegar was the most effective rinsing agent, all solutions produced lower counts after rinsing, indicating that consumers could use rinsing to remove microorganisms from chicken breast surfaces prior to cooking.

The relationship between land management, fecal indicator bacteria, and the occurrence of Campylobacter and Listeria spp. in water and sediments during synoptic sampling in the S. Fork Broad River Watershed, N.E. Georgia, U.S.A

NASA Astrophysics Data System (ADS)

Bradshaw, J. K.; Molina, M.; Sidle, R. C.; Sullivan, K.; Oakley, B.; Berrang, M.; Meinersmann, R.

2013-12-01

Fecal indicator bacteria (FIB) and pathogens stored in the bed sediments of streams and rivers may be mobilized into the water column affecting overall water quality. Furthermore, land management may play an important role in the concentrations of FIB and the occurrence of pathogens in stream water and sediments. The purpose of this study was to determine the relationship between FIB and pathogens in stream water and sediment based on three land management-affected categories: agricultural, forest, and waters receiving treated municipal wastewater. Two synoptic sampling events were conducted under baseflow conditions (<0.64 cm of rain within 24h) between October-November, 2012 and May-June, 2013. Counts of the E. coli and E. faecalis and occurrences of the enteric pathogens Campylobacter and Listeria spp. were measured in stream water and sediment samples collected at 15 locations (six agricultural (AG); six forested (FORS); and three receiving discharge from water pollution control plants (WPCP)) in the S. Fork Broad River watershed located in northeast Georgia, USA. Mean E. coli and E. faecalis concentrations were highest in the AG stream water samples (3.08 log MPN 100 mL -1 for E. coli and 3.07 log CFU 100 mL -1 for E. faecalis ) and lowest in the FORS water samples for E. coli (2.37 log MPN 100 mL -1 ) and WPCP water samples for E. faecalis (2.53 log CFU 100 mL -1 ). E. coli concentrations (2.74 log MPN 100 mL -1 ) in the WPCP streams were intermediate. Similar to water samples, E. coli concentrations were highest in the AG sediments (4.31 log MPN g -1 ), intermediate in the WPCP sediments (4.06 log MPN g -1 ), and lowest in the FORS sediments (3.46 log MPN g -1 ). In contrast to E. coli, E. faecalis concentrations were lower (1.10 to 1.31 log CFU g -1 ) and relatively more constant than E. coli in sediments over the three land management categories. Campylobacter was detected in 27% of the water samples and 8% of the sediment samples. The highest occurrence of Campylobacter detection was in the AG streams (15% of the water samples; 5% of the sediment samples). Listeria was detected in 76% of the water samples and 65% of the sediment samples. The FORS and AG streams had the highest occurrence of Listeria in water and sediment (32% and 29% of the water samples, respectively; 24% and 29% of sediment samples, respectively) suggesting Listeria is fairly ubiquitous in these streams. Based on the high concentrations of E. faecalis in water and E. coli in water and sediment, and higher frequency of Campylobacter detection in the AG streams, this study indicates that E. coli and Campylobacter may occur in high concentrations in stream sediments in land management areas where fecal material is deposited directly by livestock into the stream or adjacent land in large doses.

Quantifying the effect of hand wash duration, soap use, ground beef debris, and drying methods on the removal of Enterobacter aerogenes on hands.

PubMed

Jensen, Dane A; Danyluk, Michelle D; Harris, Linda J; Schaffner, Donald W

2015-04-01

Hand washing is recognized as a crucial step in preventing foodborne disease transmission by mitigating crosscontamination among hands, surfaces, and foods. This research was undertaken to establish the importance of several keys factors (soap, soil, time, and drying method) in reducing microorganisms during hand washing. A nonpathogenic nalidixic acid-resistant Enterobacter aerogenes surrogate for Salmonella was used to assess the efficacy of using soap or no soap for 5 or 20 s on hands with or without ground beef debris and drying with paper towel or air. Each experiment consisted of 20 replicates, each from a different individual with ∼ 6 log CFU/ml E. aerogenes on their hands. A reduction of 1.0 ± 0.4 and 1.7 ± 0.8 log CFU of E. aerogenes was observed for a 5-s wash with no soap and a 20-s wash with soap, respectively. When there was no debris on the hands, there was no significant difference between washing with and without soap for 20 s (P > 0.05). Likewise, there was no significant difference in the reductions achieved when washing without soap, whether or not debris was on the hands (P > 0.05). A significantly greater reduction (P < 0.05) in E. aerogenes (0.5 log CFU greater reduction) was observed with soap when there was ground beef debris on the hands. The greatest difference (1.1 log CFU greater average reduction) in effectiveness occurred when ground beef debris was on the hands and a 20-s wash with water was compared with a 20-s wash with soap. Significantly greater (P < 0.05) reductions were observed with paper towel drying compared with air (0.5 log CFU greater reductions). Used paper towels may contain high bacterial levels (>4.0 log CFU per towel) when hands are highly contaminated. Our results support future quantitative microbial risk assessments needed to effectively manage risks of foodborne illness in which food workers' hands are a primary cause.

Effects of the Essential Oil from Origanum vulgare L. on Survival of Pathogenic Bacteria and Starter Lactic Acid Bacteria in Semihard Cheese Broth and Slurry.

PubMed

de Souza, Geany Targino; de Carvalho, Rayssa Julliane; de Sousa, Jossana Pereira; Tavares, Josean Fechine; Schaffner, Donald; de Souza, Evandro Leite; Magnani, Marciane

2016-02-01

This study assessed the inhibitory effects of the essential oil from Origanum vulgare L. (OVEO) on Staphylococcus aureus, Listeria monocytogenes, and a mesophilic starter coculture composed of lactic acid bacteria (Lactococcus lactis subsp. lactis and L. lactis subsp. cremoris) in Brazilian coalho cheese systems. The MIC of OVEO was 2.5 μl/ml against both S. aureus and L. monocytogenes and 0.6 μl/ml against the tested starter coculture. In cheese broth containing OVEO at 0.6 μl/ml, no decrease in viable cell counts (VCC) of both pathogenic bacteria was observed, whereas the initial VCC of the starter coculture decreased approximately 1.0 log CFU/ml after 24 h of exposure at 10°C. OVEO at 1.25 and 2.5 μl/ml caused reductions of up to 2.0 and 2.5 log CFU/ml in S. aureus and L. monocytogenes, respectively, after 24 h of exposure in cheese broth. At these same concentrations, OVEO caused a greater decrease of initial VCC of the starter coculture following 4 h of exposure. Higher concentrations of OVEO were required to decrease the VCC of all target bacteria in semisolid coalho cheese slurry compared with cheese broth. The VCC of Lactococcus spp. in coalho cheese slurry containing OVEO were always lower than those of pathogenic bacteria under the same conditions. These results suggest that the concentrations of OVEO used to control pathogenic bacteria in semihard cheese should be carefully evaluated because of its inhibitory effects on the growth of starter lactic acid cultures used during the production of the product.

Curvularia haloperoxidase: antimicrobial activity and potential application as a surface disinfectant.

PubMed

Hansen, Eva H; Albertsen, Line; Schäfer, Thomas; Johansen, Charlotte; Frisvad, Jens C; Molin, Søren; Gram, Lone

2003-08-01

A presumed antimicrobial enzyme system, the Curvularia haloperoxidase system, was examined with the aim of evaluating its potential as a sanitizing agent. In the presence of hydrogen peroxide, Curvularia haloperoxidase facilitates the oxidation of halides, such as chloride, bromide, and iodide, to antimicrobial compounds. The Curvularia haloperoxidase system caused several-log-unit reductions in counts of bacteria (Pseudomonas spp., Escherichia coli, Serratia marcescens, Aeromonas salmonicida, Shewanella putrefaciens, Staphylococcus epidermidis, and Listeria monocytogenes), yeasts (Candida sp. and Rhodotorula sp.), and filamentous fungi (Aspergillus niger, Aspergillus tubigensis, Aspergillus versicolor, Fusarium oxysporum, Penicillium chrysogenum, and Penicillium paxilli) cultured in suspension. Also, bacteria adhering to the surfaces of contact lenses were killed. The numbers of S. marcescens and S. epidermidis cells adhering to contact lenses were reduced from 4.0 and 4.9 log CFU to 1.2 and 2.7 log CFU, respectively, after treatment with the Curvularia haloperoxidase system. The killing effect of the Curvularia haloperoxidase system was rapid, and 10(6) CFU of E. coli cells/ml were eliminated within 10 min of treatment. Furthermore, the antimicrobial effect was short lived, causing no antibacterial effect against E. coli 10 min after the system was mixed. Bovine serum albumin (1%) and alginate (1%) inhibited the antimicrobial activity of the Curvularia haloperoxidase system, whereas glucose and Tween 20 did not affect its activity. In conclusion, the Curvularia haloperoxidase system is an effective sanitizing system and has the potential for a vast range of applications, for instance, for disinfection of contact lenses or medical devices.

Efficacy of ozone in killing Listeria monocytogenes on alfalfa seeds and sprouts and effects on sensory quality of sprouts.

PubMed

Wade, W N; Scouten, A J; McWatters, K H; Wick, R L; Demirci, A; Fett, W F; Beuchat, L R

2003-01-01

A study was done to determine the efficacy of aqueous ozone treatment in killing Listeria monocytogenes on inoculated alfalfa seeds and sprouts. Reductions in populations of naturally occurring aerobic microorganisms on sprouts and changes in the sensory quality of sprouts were also determined. The treatment (10 or 20 min) of seeds in water (4 degrees C) containing an initial concentration of 21.8 +/- 0.1 microg/ml of ozone failed to cause a significant (P < or = 0.05) reduction in populations of L. monocytogenes. The continuous sparging of seeds with ozonated water (initial ozone concentration of 21.3 +/- 0.2 microg/ml) for 20 min significantly reduced the population by 1.48 log10 CFU/g. The treatment (2 min) of inoculated alfalfa sprouts with water containing 5.0 +/- 0.5, 9.0 +/- 0.5, or 23.2 +/- 1.6 microg/ml of ozone resulted in significant (P < or = 0.05) reductions of 0.78, 0.81, and 0.91 log10 CFU/g, respectively, compared to populations detected on sprouts treated with water. Treatments (2 min) with up to 23.3 +/- 1.6 microg/ml of ozone did not significantly (P > 0.05) reduce populations of aerobic naturally occurring microorganisms. The continuous sparging of sprouts with ozonated water for 5 to 20 min caused significant reductions in L. monocytogenes and natural microbiota compared to soaking in water (control) but did not enhance the lethality compared to the sprouts not treated with continuous sparging. The treatment of sprouts with ozonated water (20.0 microg/ml) for 5 or 10 min caused a significant deterioration in the sensory quality during subsequent storage at 4 degrees C for 7 to 11 days. Scanning electron microscopy of uninoculated alfalfa seeds and sprouts showed physical damage, fungal and bacterial growth, and biofilm formation that provide evidence of factors contributing to the difficulty of killing microorganisms by treatment with ozone and other sanitizers.

Bifidobacterium pseudolongum are efficient indicators of animal fecal contamination in raw milk cheese industry

PubMed Central

2011-01-01

Background The contamination of raw milk cheeses (St-Marcellin and Brie) from two plants in France was studied at several steps of production (raw milk, after addition of rennet - St-Marcellin - or after second maturation - Brie -, after removal from the mold and during ripening) using bifidobacteria as indicators of fecal contamination. Results Bifidobacterium semi-quantitative counts were compared using PCR-RFLP and real-time PCR. B. pseudolongum were detected in 77% (PCR-RFLP; 1.75 to 2.29 log cfu ml-1) and 68% (real-time PCR; 2.19 to 2.73 log cfu ml-1) of St-Marcellin samples and in 87% (PCR-RFLP; 1.17 to 2.40 log cfu ml-1) of Brie cheeses samples. Mean counts of B. pseudolongum remained stable along both processes. Two other populations of bifidobacteria were detected during the ripening stage of St-Marcellin, respectively in 61% and 18% of the samples (PCR-RFLP). The presence of these populations explains the increase in total bifidobacteria observed during ripening. Further characterization of these populations is currently under process. Forty-eight percents (St-Marcellin) and 70% (Brie) of the samples were B. pseudolongum positive/E. coli negative while only 10% (St-Marcellin) and 3% (Brie) were B. pseudolongum negative/E. coli positive. Conclusions The increase of total bifidobacteria during ripening in Marcellin's process does not allow their use as fecal indicator. The presence of B. pseudolongum along the processes defined a contamination from animal origin since this species is predominant in cow dung and has never been isolated in human feces. B. pseudolongum was more sensitive as an indicator than E. coli along the two different cheese processes. B. pseudolongum should be used as fecal indicator rather than E. coli to assess the quality of raw milk and raw milk cheeses. PMID:21816092

Evaluation of a Disinfectant Wipe Intervention on Fomite-to-Finger Microbial Transfer

PubMed Central

Lopez, Gerardo U.; Kitajima, Masaaki; Havas, Aaron; Gerba, Charles P.

2014-01-01

Inanimate surfaces, or fomites, can serve as routes of transmission of enteric and respiratory pathogens. No previous studies have evaluated the impact of surface disinfection on the level of pathogen transfer from fomites to fingers. Thus, the present study investigated the change in microbial transfer from contaminated fomites to fingers following disinfecting wipe use. Escherichia coli (108 to 109 CFU/ml), Staphylococcus aureus (109 CFU/ml), Bacillus thuringiensis spores (107 to 108 CFU/ml), and poliovirus 1 (108 PFU/ml) were seeded on ceramic tile, laminate, and granite in 10-μl drops and allowed to dry for 30 min at a relative humidity of 15 to 32%. The seeded fomites were treated with a disinfectant wipe and allowed to dry for an additional 10 min. Fomite-to-finger transfer trials were conducted to measure concentrations of transferred microorganisms on the fingers after the disinfectant wipe intervention. The mean log10 reduction of the test microorganisms on fomites by the disinfectant wipe treatment varied from 1.9 to 5.0, depending on the microorganism and the fomite. Microbial transfer from disinfectant-wipe-treated fomites was lower (up to <0.1% on average) than from nontreated surfaces (up to 36.3% on average, reported in our previous study) for all types of microorganisms and fomites. This is the first study quantifying microbial transfer from contaminated fomites to fingers after the use of disinfectant wipe intervention. The data generated in the present study can be used in quantitative microbial risk assessment models to predict the effect of disinfectant wipes in reducing microbial exposure. PMID:24610856

GFP tagged Vibrio parahaemolyticus Dahv2 infection and the protective effects of the probiotic Bacillus licheniformis Dahb1 on the growth, immune and antioxidant responses in Pangasius hypophthalmus.

PubMed

Gobi, Narayanan; Malaikozhundan, Balasubramanian; Sekar, Vijayakumar; Shanthi, Sathappan; Vaseeharan, Baskaralingam; Jayakumar, Rengarajan; Khudus Nazar, Abdul

2016-05-01

In this study, the pathogenicity of GFP tagged Vibrio parahaemolyticus Dahv2 and the protective effect of the probiotic strain, Bacillus licheniformis Dahb1 was studied on the Asian catfish, Pangasius hypophthalmus. The experiment was carried out for 24 days with three groups and one group served as the control (without treatment). In the first group, P. hypophthalmus was orally infected with 1 mL of GFP tagged V. parahaemolyticus Dahv2 at two different doses (10(5) and 10(7) cfu mL(-1)). In the second group, P. hypophthalmus was orally administrated with 1 ml of the probiotic B. licheniformis Dahb1 at two different doses (10(5) and 10(7) cfu mL(-1)). In the third group, P. hypophthalmus was orally infected first with 1 mL of GFP tagged V. parahaemolyticus Dahv2 followed by the administration of 1 mL of B. licheniformis Dahb1 (combined treatment) at two different doses (10(5) and 10(7) cfu mL(-1)). The growth, immune (myeloperoxidase, respiratory burst, natural complement haemolytic and lysozyme activity) and antioxidant (glutathione-S-transferase, reduced glutathione and total glutathione) responses of P. hypophthalmus were reduced after post infection of GFP tagged V. parahaemolyticus Dahv2 compared to control. However, after administration with the probiotic B. licheniformis Dahb1 at 10(5) cfu mL(-1), P. hypophthalmus showed significant increase in the growth, immune and antioxidant responses compared to 10(7) cfu mL(-1). On the otherhand, the growth, immune and antioxidant responses of P. hypophthalmus infected and administrated with combined GFP tagged Vibrio + Bacillus at 10(5) cfu mL(-1) were relatively higher than that of GFP tagged V. parahaemolyticus Dahv2 and control groups but lower than that of probiotic B. licheniformis Dahb1 groups. The results of the present study conclude that the probiotic B. licheniformis Dahb1 at 10(5) cfu mL(-1) has the potential to protect the P. hypophthalmus against V. parahaemolyticus Dahv2 infection by enhancing the growth, immune and antioxidant responses. The probiotic B. licheniformis Dahb1 would be effectively used in the treatment of aquatic diseases for improvement of aquaculture industry. Copyright © 2016 Elsevier Ltd. All rights reserved.

Inactivation of enterohemorrhagic Escherichia coli in rumen content- or feces-contaminated drinking water for cattle.

PubMed

Zhao, Tong; Zhao, Ping; West, Joe W; Bernard, John K; Cross, Heath G; Doyle, Michael P

2006-05-01

Cattle drinking water is a source of on-farm Escherichia coli O157:H7 transmission. The antimicrobial activities of disinfectants to control E. coli O157:H7 in on-farm drinking water are frequently neutralized by the presence of rumen content and manure that generally contaminate the drinking water. Different chemical treatments, including lactic acid, acidic calcium sulfate, chlorine, chlorine dioxide, hydrogen peroxide, caprylic acid, ozone, butyric acid, sodium benzoate, and competing E. coli, were tested individually or in combination for inactivation of E. coli O157:H7 in the presence of rumen content. Chlorine (5 ppm), ozone (22 to 24 ppm at 5 degrees C), and competing E. coli treatment of water had minimal effects (<1 log CFU/ml reduction) on killing E. coli O157:H7 in the presence of rumen content at water-to-rumen content ratios of 50:1 (vol/wt) and lower. Four chemical-treatment combinations, including (i) 0.1% lactic acid, 0.9% acidic calcium sulfate, and 0.05% caprylic acid (treatment A); (ii) 0.1% lactic acid, 0.9% acidic calcium sulfate, and 0.1% sodium benzoate (treatment B); (iii) 0.1% lactic acid, 0.9% acidic calcium sulfate, and 0.5% butyric acid (treatment C); and (iv) 0.1% lactic acid, 0.9% acidic calcium sulfate, and 100 ppm chlorine dioxide (treatment D); were highly effective (>3 log CFU/ml reduction) at 21 degrees C in killing E. coli O157:H7, O26:H11, and O111:NM in water heavily contaminated with rumen content (10:1 water/rumen content ratio [vol/wt]) or feces (20:1 water/feces ratio [vol/wt]). Among them, treatments A, B, and C killed >5 log CFU E. coli O157:H7, O26:H11, and O111:NM/ml within 30 min in water containing rumen content or feces, whereas treatment D inactivated approximately 3 to 4 log CFU/ml under the same conditions. Cattle given water containing treatment A or C or untreated water (control) ad libitum for two 7-day periods drank 15.2, 13.8, and 30.3 liters/day, respectively, and cattle given water containing 0.1% lactic acid plus 0.9% acidic calcium sulfate (pH 2.1) drank 18.6 liters/day. The amounts of water consumed for all water treatments were significantly different from that for the control, but there were no significant differences among the water treatments. Such treatments may best be applied periodically to drinking water troughs and then flushed, rather than being added continuously, to avoid reduced water consumption by cattle.

Optimization of Culture Medium for the Growth of Candida sp. and Blastobotrys sp. as Starter Culture in Fermentation of Cocoa Beans (Theobroma cacao) Using Response Surface Methodology (RSM).

PubMed

Mahazar, N H; Zakuan, Z; Norhayati, H; MeorHussin, A S; Rukayadi, Y

2017-01-01

Inoculation of starter culture in cocoa bean fermentation produces consistent, predictable and high quality of fermented cocoa beans. It is important to produce healthy inoculum in cocoa bean fermentation for better fermented products. Inoculum could minimize the length of the lag phase in fermentation. The purpose of this study was to optimize the component of culture medium for the maximum cultivation of Candida sp. and Blastobotrys sp. Molasses and yeast extract were chosen as medium composition and Response Surface Methodology (RSM) was then employed to optimize the molasses and yeast extract. Maximum growth of Candida sp. (7.63 log CFU mL-1) and Blastobotrys sp. (8.30 log CFU mL-1) were obtained from the fermentation. Optimum culture media for the growth of Candida sp., consist of 10% (w/v) molasses and 2% (w/v) yeast extract, while for Blastobotrys sp., were 1.94% (w/v) molasses and 2% (w/v) yeast extract. This study shows that culture medium consists of molasses and yeast extract were able to produce maximum growth of Candida sp. and Blastobotrys sp., as a starter culture for cocoa bean fermentation.

Inactivation of Escherichia coli O157:H7 attached to spinach harvester blade using bacteriophage.

PubMed

Patel, Jitendra; Sharma, Manan; Millner, Patricia; Calaway, Todd; Singh, Manpreet

2011-04-01

Outbreaks associated with leafy greens have focused attention on the transfer of human pathogens to these commodities during harvest with commercial equipment. Attachment of Escherichia coli O157:H7 on new or rusty spinach harvester blades immersed in spinach extract or 10% tryptic soy broth (TSB) was investigated. Bacteriophages specific for E. coli O157:H7 were evaluated to kill cells attached to blade. A cocktail of five nalidixic acid-resistant E. coli O157:H7 isolates was transferred to 25 mL of spinach extract or 10% TSB. A piece of sterilized spinach harvester blade (2×1") was placed in above spinach extract or 10% TSB and incubated at room (22 °C) or dynamic (30 °C day, 20 °C night) temperatures. E. coli O157:H7 populations attached to blade during incubation in spinach extract or 10% TSB were determined. When inoculated at 1 log CFU/mL, E. coli O157:H7 attachment to blades after 24 and 48 h incubation at dynamic temperature (6.09 and 6.37 log CFU/mL) was significantly higher than when incubated at 22 °C (4.84 and 5.68 log CFU/mL), respectively. After 48 h incubation, two blades were sprayed on each side with a cocktail of E. coli O157-specific bacteriophages before scraping the blade, and subsequent plating on Sorbitol MacConkey media-nalidixic acid. Application of bacteriophages reduced E. coli O157:H7 populations by 4.5 log CFU on blades after 2 h of phage treatment. Our study demonstrates that E. coli O157:H7 can attach to and proliferate on spinach harvester blades under static and dynamic temperature conditions, and bacteriophages are able to reduce E. coli O157:H7 populations adhered to blades. © Mary Ann Liebert, Inc.

Pneumococcal LytA Autolysin, a Potent Therapeutic Agent in Experimental Peritonitis-Sepsis Caused by Highly β-Lactam-Resistant Streptococcus pneumoniae▿

PubMed Central

Rodríguez-Cerrato, Violeta; García, Pedro; Huelves, Lorena; García, Ernesto; del Prado, Gema; Gracia, Matilde; Ponte, Carmen; López, Rubens; Soriano, Francisco

2007-01-01

The in vitro and in vivo antipneumococcal activities of the main pneumococcal autolysin (LytA) and Cpl-1, a lysozyme encoded by phage Cp-1, were studied. Intraperitoneal therapy with LytA or high-dose Cpl-1 remarkably reduced peritoneal bacterial counts (>5 log10 CFU/ml) compared with those for the controls. After intravenous injection, LytA was the most effective treatment. PMID:17576844

Bacterial recovery from breast skin of genetically feathered and featherless broiler carcasses immediately following scalding and picking.

PubMed

Buhr, R J; Berrang, M E; Cason, J A

2003-10-01

Genetically feathered and featherless sibling broilers selected for matched BW were killed, scalded, and defeathered to determine the consequences of feathers and empty feather follicles on the recovery of bacteria from carcass breast skin. In trial 1, the vents of all carcasses were plugged and sutured before scalding to prevent the expulsion of cloacal contents during picking. In trial 2, half of the carcasses had their vents plugged and sutured. Immediately after defeathering, breast skin was aseptically removed, and bacteria associated with it were enumerated. In trial 1, the levels of bacteria recovered did not differ between feathered and featherless carcasses: Campylobacter log10 1.4 cfu/mL of rinse, coliform log10 1.8, Escherichia coli log10 1.6, and total aerobic bacteria log10 3.1. In trial 2, the carcasses that had vents plugged and sutured had lower levels of all four types of bacteria (differences of Campylobacter log10 0.7 cfu/mL, coliform log10 1.8, E. coli log10 1.7, and total aerobic bacteria log10 0.5) than those carcasses with open vents. The lower levels of bacteria recovered from carcasses with the vents plugged and sutured during picking enabled detection of small but significant differences between feathered and featherless carcasses. The level of coliform and E. coli recovered was slightly higher by log10 0.7 cfu for feathered carcasses, but featherless carcasses had marginally higher levels of total aerobic bacteria by log10 0.4 cfu. Feathered and featherless carcasses with open vents during picking did not differ in the levels of recovery of coliform, E. coli, and total aerobic bacteria from breast skin.

Survival of the probiotic, L. plantarum 299v and its effects on the faecal bacterial flora, with and without gastric acid inhibition.

PubMed

Goossens, D; Jonkers, D; Russel, M; Thijs, A; van den Bogaard, A; Stobberingh, E; Stockbrügger, R

2005-01-01

Probiotic bacteria have to survive passage through the gastrointestinal tract. In this placebo-controlled double-blind study, the effect of Lactobacillus plantarum 299v on the faecal flora was studied with and without gastric acid inhibition. Thirty-two healthy volunteers were given pantoprazole (40 mg/day) or placebo for 3 weeks from week 2 until week 4. In addition, from week 3 until week 4, L. plantarum 299v in an oatmeal-fermented drink (10(9) CFU/ml) was given twice daily to both groups. From each healthy volunteer, faecal samples were collected at the end of week 1, 2, 4 and 8 (4 weeks after cessation of L. plantarum 299v and pantoprazole/placebo). Several aerobically and anaerobically growing bacteria were counted and short chain fatty acid concentrations were determined. In both the pantoprazole and the placebo group, median lactobacilli counts increased significantly in week 4 compared to week 1 (from log 4.5 to 8.0 CFU/g faeces in pantoprazole and from log 4.2 to 7.7 CFU/g faeces in placebo group) and decreased significantly in week 8 (to log 4.5 CFU/g faeces in pantoprazole and log 4.3 CFU/g faeces in placebo group). These lactobacilli were identified as L. plantarum 299v. No significant differences were observed in all other bacterial counts and short chain fatty acid concentrations. The comparable increase of faecal lactobacilli counts in both the pantoprazole and the placebo-treated group demonstrates that L. plantarum 299v survives passage through the gastrointestinal tract irrespective of gastric acidity. The increment of the intra-gastric pH in combination with L. plantarum 299v did not modulate bacterial composition and/or the production of short chain fatty acids.

Bactericidal Effect and Pharmacodynamics of Cethromycin (ABT-773) in a Murine Pneumococcal Pneumonia Model

PubMed Central

Kim, Myo-Kyoung; Zhou, Wen; Tessier, Pamela R.; Xuan, Dawei; Ye, Min; Nightingale, Charles H.; Nicolau, David P.

2002-01-01

Cethromycin (ABT-773), a new ketolide, possesses potent in vitro activity against Streptococcus pneumoniae. The objective of this study was to investigate the in vivo bactericidal activity of cethromycin against macrolide-susceptible and -resistant S. pneumoniae in a murine pneumonia model and to describe the pharmacodynamic (PD) profile of cethromycin. Eight (two macrolide susceptible, six macrolide resistant) clinical isolates of S. pneumoniae were investigated. Cyclophosphamide administration rendered ICR mice transiently neutropenic prior to intratracheal inoculation with 0.05 ml of an S. pneumoniae suspension containing 107 to 108 CFU/ml. Oral cethromycin was initiated 12 to 14 h postinoculation over a dosage range of 0.1 to 800 mg/kg of body weight/day. Lungs from seven to eight mice per treatment and control groups were collected at 0 and 24 h posttherapy to assess bacterial density. The cumulative mortality (n = 12 to 13) was assessed at 120 h (end of therapy) and at 192 h (3 days posttherapy). Recovery of pneumococci from the lungs of infected animals prior to the initiation of therapy ranged from 4.6 to 7.2 log10 CFU. Growth in untreated control animals over a 24-h study period increased 0.3 to 2.7 log10 CFU. Cethromycin demonstrated a substantial bactericidal effect, regardless of macrolide susceptibility. Correlation between changes in bacterial density (24 h) and survival over both 120 and 192 h were statistically significant. All three PD parameters demonstrated a significant correlation with changes in log10 CFU/lung (Spearman's correlation coefficient, P < 0.001); however, the goodness of fit as assessed with the maximum effect (Emax) model revealed that the maximum concentration of free drug in serum (Cmax free)/MIC and the area under the free drug concentration-time curve (AUCfree)/MIC best explained the relationship between drug exposure and reductions in viable bacterial counts. These data reveal that an approximate cethromycin AUCfree/MIC of 50 or Cmax free/MIC of 1 results in bacteriostatic effects, while higher values (twofold) maximize survival. PMID:12234843

Survival or growth of inoculated Escherichia coli O157:H7 and Salmonella on yellow onions (Allium cepa) under conditions simulating food service and consumer handling and storage.

PubMed

Lieberman, Vanessa M; Zhao, Irene Y; Schaffner, Donald W; Danyluk, Michelle D; Harris, Linda J

2015-01-01

Whole and diced yellow onions (Allium cepa) were inoculated with five-strain cocktails of rifampin-resistant Escherichia coli O157:H7 or Salmonella and stored under conditions to simulate food service or consumer handling. The inoculum was grown in broth (for both whole and diced onion experiments) or on agar plates (for whole onion experiments). Marked circles (3.3 cm in diameter) on the outer papery skin of whole onions were spot inoculated (10 μl in 10 drops) at 7 log CFU per circle, and onions were stored at 4°C, 30 to 50 % relative humidity, or at ambient conditions (23°C, 30 to 50 % relative humidity). Diced onions were inoculated at 3 log CFU/g and then stored in open or closed containers at 4°C or ambient conditions. Previously inoculated and ambient-stored diced onions were also mixed 1:9 (wt/wt) with refrigerated uninoculated freshly diced onions and stored in closed containers at ambient conditions. Inoculated pathogens were recovered in 0.1 % peptone and plated onto selective and nonselective media supplemented with 50 μg/ml rifampin. Both E. coli O157:H7 and Salmonella populations declined more rapidly on onion skins when the inoculum was prepared in broth rather than on agar. Agar-prepared E. coli O157:H7 and Salmonella declined by 0.4 and 0.3 log CFU per sample per day, respectively, at ambient conditions; at 4°C the rates of reduction were 0.08 and 0.06 log CFU per sample per day for E. coli O157:H7 and Salmonella, respectively. Populations of E. coli O157:H7 and Salmonella did not change over 6 days of storage at 4°C in diced onions. Lag times of 6 to 9 h were observed with freshly inoculated onion at ambient conditions; no lag was observed when previously inoculated and uninoculated onions were mixed. Growth rates at ambient conditions were 0.2 to 0.3 log CFU/g/h for E. coli O157:H7 and Salmonella in freshly inoculated onion and 0.2 log CFU/g/h in mixed product. Diced onions support pathogen growth and should be kept refrigerated.

Industrial Cooling Tower Disinfection Treatment to Prevent Legionella spp.

PubMed

Iervolino, Matteo; Mancini, Benedetta; Cristino, Sandra

2017-09-26

The contamination of industrial cooling towers has been identified as one cause of legionellosis, but the real risk has been underestimated. Two different disinfection treatments were tested on Legionella colonization in an industrial Cooling Tower System (CTS). Environmental monitoring of Legionella , P. aeruginosa , and a heterotrophic plate count (HPC) at 36 °C was performed from June to October 2016. The disinfection procedures adopted were based on hydrogen peroxide (H₂O₂) and silver salts (Ag⁺), in addition to an anti-algal treatment, then using hyperclorination as a shock, and then continuous treatment by sodium hypochlorite (NaClO). L . pneumophila serogroup 8 was found at a concentration of 5.06 Log cfu/L after the CTS filling; a shock treatment performed by H₂O₂/Ag⁺ produced a rapid increase in contamination up to 6.14 Log cfu/L. The CTS activity was stopped and two subsequent shock treatments were performed using NaClO, followed by continuous hyperclorination. These procedures showed a significant decrease ( p < 0.05) in Legionella concentration (1.77 Log cfu/L). The same trend was observed for P . aeruginosa (0.55 Log cfu/100 mL) and HPC (1.95 Log cfu/mL) at 36 °C. Environmental monitoring and the adoption of maintenance procedures, including anti-scale treatment, and physical, chemical, and microbiological control, ensure the good performance of a CTS, reducing the Legionella risk for public health.

Effectiveness of liquid soap vs. chlorhexidine gluconate for the removal of Clostridium difficile from bare hands and gloved hands.

PubMed

Bettin, K; Clabots, C; Mathie, P; Willard, K; Gerding, D N

1994-11-01

To compare liquid soap versus 4% chlorhexidine gluconate in 4% alcohol for the decontamination of bare or gloved hands inoculated with an epidemic strain of Clostridium difficile. C difficile (6.7 log10 colony-forming units [CFU], 47% spores), was seeded onto bare or latex gloved hands of ten volunteers and allowed to dry. Half the volunteers initially washed with soap and half with chlorhexidine, followed by the other agent 1 week later. Cultures were done with Rodac plates at three sites on the hand: finger/thumbtips, the palmar surfaces of the fingers, and the palm. Statistical comparison was by paired Student's t test. On bare hands, soap and chlorhexidine did not differ in residual bacterial counts on the finger/thumbtips (log10 CFU, 2.0 and 2.1, P = NS) and fingers (log10 CFU, 2.4 and 2.5, P = NS). Counts were too high on bare palms to quantitate. On gloved hands, soap was more effective than chlorhexidine on fingers (log10 CFU 1.3 and 1.7, P < .01) and palms (log10 CFU 1.5 and 2.0, P < .01), but not finger/thumbtips (log10 CFU 1.6 with each, P = NS). Residual C difficile counts were lower on gloved hands than bare hands (P < 0.01 to < 0.0001). The two agents did not differ significantly in residual counts of C difficile on bare hands, but on gloved hands residual counts were lower following soap wash than following chlorhexidine wash. These observations support the use of either soap or chlorhexidine as a handwash for removal of C difficile, but efficacy in the prevention of C difficile transmission must be determined by prospective clinical trials.

Atmospheric pressure plasma jet for bacterial decontamination and property improvement of fruit and vegetable processing wastewater

NASA Astrophysics Data System (ADS)

Mohamed, Abdel-Aleam H.; Shariff, Samir M. Al; Ouf, Salama A.; Benghanem, Mohamed

2016-05-01

An atmospheric pressure plasma jet was tested for decontaminating and improving the characteristics of wastewater derived from blackberry, date palm, tomato and beetroot processing industries. The jet was generated by blowing argon gas through a cylindrical alumina tube while a high voltage was applied between two electrodes surrounding the tube. Oxygen gas was mixed with argon at the rate of 0.2% and the argon mass flow was fixed at 4.5 slm. Images show that the generated plasma jet penetrated the treated wastewater samples. Plasma emission spectra show the presence of O and OH radicals as well as excited molecular nitrogen and argon. Complete decontamination of wastewater derived from date palm and tomato processing was achieved after 120 and 150 s exposure to the plasma jet, respectively. The bacterial count of wastewater from blackberry and beetroot was reduced by 0.41 and 2.24 log10 colony-forming units (CFU) per ml, respectively, after 180 s. Escherichia coli was the most susceptible bacterial species to the cold plasma while Shigella boydii had the minimum susceptibility, recording 1.30 and 3.34 log10 CFU ml-1, respectively, as compared to the 7.00 log10 initial count. The chemical oxygen demands of wastewater were improved by 57.5-93.3% after 180 s exposure to the plasma jet being tested. The endotoxins in the wastewater were reduced by up to 90.22%. The variation in plasma effectiveness is probably related to the antioxidant concentration of the different investigated wastewaters.

Sanitizing in Dry-Processing Environments Using Isopropyl Alcohol Quaternary Ammonium Formula.

PubMed

Kane, Deborah M; Getty, Kelly J K; Mayer, Brian; Mazzotta, Alejandro

2016-01-01

Dry-processing environments are particularly challenging to clean and sanitize because introduced water can favor growth and establishment of pathogenic microorganisms such as Salmonella. Our objective was to determine the efficacy of an isopropyl alcohol quaternary ammonium (IPAQuat) formula for eliminating potential Salmonella contamination on food contact surfaces. Clean stainless steel coupons and conveyor belt materials used in dry-processing environments were spot inoculated in the center of coupons (5 by 5 cm) with a six-serotype composite of Salmonella (approximately 10 log CFU/ml), subjected to IPAQuat sanitizer treatments with exposure times of 30 s, 1 min, or 5 min, and then swabbed for enumeration of posttreatment survivors. A subset of inoculated surfaces was soiled with a breadcrumb-flour blend and allowed to sit on the laboratory bench for a minimum of 16 h before sanitation. Pretreatment Salmonella populations (inoculated controls, 0 s treatment) were approximately 7.0 log CFU/25 cm(2), and posttreatment survivors were 1.31, 0.72, and < 0.7 (detection limit) log CFU/25 cm(2) after sanitizer exposure for 30 s, 1 min, or 5 min, respectively, for both clean (no added soil) and soiled surfaces. Treatment with the IPAQuat formula using 30-s sanitizer exposures resulted in 5.68-log reductions, whereas >6.0-log reductions were observed for sanitizer exposures of 1 and 5 min. Because water is not introduced into the processing environment with this approach, the IPAQuat formula could have sanitation applications in dry-processing environments to eliminate potential contamination from Salmonella on food contact surfaces.

Attachment of and biofilm formation by Enterobacter sakazakii on stainless steel and enteral feeding tubes.

PubMed

Kim, Hoikyung; Ryu, Jee-Hoon; Beuchat, Larry R

2006-09-01

Enterobacter sakazakii has been reported to form biofilms, but environmental conditions affecting attachment to and biofilm formation on abiotic surfaces have not been described. We did a study to determine the effects of temperature and nutrient availability on attachment and biofilm formation by E. sakazakii on stainless steel and enteral feeding tubes. Five strains grown to stationary phase in tryptic soy broth (TSB), infant formula broth (IFB), or lettuce juice broth (LJB) at 12 and 25 degrees C were examined for the extent to which they attach to these materials. Higher populations attached at 25 degrees C than at 12 degrees C. Stainless steel coupons and enteral feeding tubes were immersed for 24 h at 4 degrees C in phosphate-buffered saline suspensions (7 log CFU/ml) to facilitate the attachment of 5.33 to 5.51 and 5.03 to 5.12 log CFU/cm(2), respectively, before they were immersed in TSB, IFB, or LJB, followed by incubation at 12 or 25 degrees C for up to 10 days. Biofilms were not produced at 12 degrees C. The number of cells of test strains increased by 1.42 to 1.67 log CFU/cm(2) and 1.16 to 1.31 log CFU/cm(2) in biofilms formed on stainless steel and feeding tubes, respectively, immersed in IFB at 25 degrees C; biofilms were not formed on TSB and LJB at 25 degrees C, indicating that nutrient availability plays a major role in processes leading to biofilm formation on the surfaces of these inert materials. These observations emphasize the importance of temperature control in reconstituted infant formula preparation and storage areas in preventing attachment and biofilm formation by E. sakazakii.

Curvularia Haloperoxidase: Antimicrobial Activity and Potential Application as a Surface Disinfectant

PubMed Central

Hansen, Eva H.; Albertsen, Line; Schäfer, Thomas; Johansen, Charlotte; Frisvad, Jens C.; Molin, Søren; Gram, Lone

2003-01-01

A presumed antimicrobial enzyme system, the Curvularia haloperoxidase system, was examined with the aim of evaluating its potential as a sanitizing agent. In the presence of hydrogen peroxide, Curvularia haloperoxidase facilitates the oxidation of halides, such as chloride, bromide, and iodide, to antimicrobial compounds. The Curvularia haloperoxidase system caused several-log-unit reductions in counts of bacteria (Pseudomonas spp., Escherichia coli, Serratia marcescens, Aeromonas salmonicida, Shewanella putrefaciens, Staphylococcus epidermidis, and Listeria monocytogenes), yeasts (Candida sp. and Rhodotorula sp.), and filamentous fungi (Aspergillus niger, Aspergillus tubigensis, Aspergillus versicolor, Fusarium oxysporum, Penicillium chrysogenum, and Penicillium paxilli) cultured in suspension. Also, bacteria adhering to the surfaces of contact lenses were killed. The numbers of S. marcescens and S. epidermidis cells adhering to contact lenses were reduced from 4.0 and 4.9 log CFU to 1.2 and 2.7 log CFU, respectively, after treatment with the Curvularia haloperoxidase system. The killing effect of the Curvularia haloperoxidase system was rapid, and 106 CFU of E. coli cells/ml were eliminated within 10 min of treatment. Furthermore, the antimicrobial effect was short lived, causing no antibacterial effect against E. coli 10 min after the system was mixed. Bovine serum albumin (1%) and alginate (1%) inhibited the antimicrobial activity of the Curvularia haloperoxidase system, whereas glucose and Tween 20 did not affect its activity. In conclusion, the Curvularia haloperoxidase system is an effective sanitizing system and has the potential for a vast range of applications, for instance, for disinfection of contact lenses or medical devices. PMID:12902249

Efficacy of chlorine, acidic electrolyzed water and aqueous chlorine dioxide solutions to decontaminate Escherichia coli O157:H7 from lettuce leaves.

PubMed

Keskinen, Lindsey A; Burke, Angela; Annous, Bassam A

2009-06-30

This study compared the efficacy of chlorine (20-200 ppm), acidic electrolyzed water (50 ppm chlorine, pH 2.6), acidified sodium chlorite (20-200 ppm chlorite ion concentration, Sanova), and aqueous chlorine dioxide (20-200 ppm chlorite ion concentration, TriNova) washes in reducing populations of Escherichia coli O157:H7 on artificially inoculated lettuce. Fresh-cut leaves of Romaine or Iceberg lettuce were inoculated by immersion in water containing E. coli O157:H7 (8 log CFU/ml) for 5 min and dried in a salad spinner. Leaves (25 g) were then washed for 2 min, immediately or following 24 h of storage at 4 degrees C. The washing treatments containing chlorite ion concentrations of 100 and 200 ppm were the most effective against E. coli O157:H7 populations on Iceberg lettuce, with log reductions as high as 1.25 log CFU/g and 1.05 log CFU/g for TriNova and Sanova wash treatments, respectively. All other wash treatments resulted in population reductions of less than 1 log CFU/g. Chlorine (200 ppm), TriNova, Sanova, and acidic electrolyzed water were all equally effective against E. coli O157:H7 on Romaine, with log reductions of approximately 1 log CFU/g. The 20 ppm chlorine wash was as effective as the deionized water wash in reducing populations of E. coli O157:H7 on Romaine and Iceberg lettuce. Scanning electron microscopy indicated that E. coli O157:H7 that was incorporated into biofilms or located in damage lettuce tissue remained on the lettuce leaf, while individual cells on undamaged leaf surfaces were more likely to be washed away.

Microbiological quality of drinking water from dispensers in roadside restaurants of Bangladesh.

PubMed

Moniruzzaman, M; Akter, S; Islam, M A; Mia, Z

2011-01-15

The microbiological status of water from dispensers in different roadside restaurants of Dhaka city and Savar area was analyzed in this study. Seven samples from Dhaka and 8 samples of Savar were checked. The heterotrophic plate count was in a range of 1.0 x 10(3) CFU mL(-1) to 2.0 x 10(4) CFU mL(-1) (from new bottles), 1.0 x 10(3) to 1.5 x 10(4) CFU mL(-1) (after dispensation), and 1.5 x 10(3) CFU mL(-1) to 1.0 x l0(5) CFU mL(-1) (from serving glass). In several of the samples, the heterotrophic plate count was higher than the count in water from new bottle or after dispensation, suggesting added contamination from the serving glass. 80% of the samples were contaminated with total and fecal coliform bacteria, which render these waters unacceptable for human consumption. The samples were found to contain gram negative bacteria like E coli, Shigella sp., Klebsiella sp., Enterobacter sp., Pseudomonas sp., and Salmonella sp., which are potential pathogens and thus pose a serious threat to public health. This study elucidates the importance of monitoring the bottling companies and the restaurants and put them under strict regulations to prevent future outbreak of any water borne diseases caused by consumption of dispensed water.

Performance characteristics and estimation of measurement uncertainty of three plating procedures for Campylobacter enumeration in chicken meat.

PubMed

Habib, I; Sampers, I; Uyttendaele, M; Berkvens, D; De Zutter, L

2008-02-01

In this work, we present an intra-laboratory study in order to estimate repeatability (r), reproducibility (R), and measurement uncertainty (U) associated with three media for Campylobacter enumeration, named, modified charcoal cefoperazone deoxycholate agar (mCCDA); Karmali agar; and CampyFood ID agar (CFA) a medium by Biomérieux SA. The study was performed at three levels: (1) pure bacterial cultures, using three Campylobacter strains; (2) artificially contaminated samples from three chicken meat matrixes (total n=30), whereby samples were spiked using two contamination levels; ca. 10(3)cfuCampylobacter/g, and ca. 10(4)cfuCampylobacter/g; and (3) pilot testing in naturally contaminated chicken meat samples (n=20). Results from pure culture experiment revealed that enumeration of Campylobacter colonies on Karmali and CFA media was more convenient in comparison with mCCDA using spread and spiral plating techniques. Based on artificially contaminated samples testing, values of repeatability (r) were comparable between the three media, and estimated as 0.15log(10)cfu/g for mCCDA, 0.14log(10)cfu/g for Karmali, and 0.18log(10)cfu/g for CFA. As well, reproducibility performance of the three plating media was comparable. General R values which can be used when testing chicken meat samples are; 0.28log(10), 0.32log(10), and 0.25log(10) for plating on mCCDA, Karmali agar, and CFA, respectively. Measurement uncertainty associated with mCCDA, Karmali agar, and CFA using spread plating, for combination of all meat matrixes, were +/-0.24log(10)cfu/g, +/-0.28log(10)cfu/g, and +/-0.22log(10)cfu/g, respectively. Higher uncertainty was associated with Karmali agar for Campylobacter enumeration in artificially inoculated minced meat (+/-0.48log(10)cfu/g). The general performance of CFA medium was comparable with mCCDA performance at the level of artificially contaminated samples. However, when tested at naturally contaminated samples, non-Campylobacter colonies gave similar deep red colour as that given by the typical Campylobacter growth on CFA. Such colonies were not easily distinguishable by naked eye. In general, the overall reproducibility, repeatability, and measurement uncertainty estimated by our study indicate that there are no major problems with the precision of the International Organization for Standardization (ISO) 10272-2:2006 protocol for Campylobacter enumeration using mCCDA medium.

Antibacterial TAP-mimic electrospun polymer scaffold: effects on P. gingivalis-infected dentin biofilm.

PubMed

Albuquerque, Maria Tereza P; Evans, Joshua D; Gregory, Richard L; Valera, Marcia C; Bottino, Marco C

2016-03-01

This study sought to investigate, in vitro, the effects of a recently developed triple antibiotic paste (TAP)-mimic polymer nanofibrous scaffold against Porphyromonas gingivalis-infected dentin biofilm. Dentin specimens (4 × 4 × 1 mm(3)) were prepared from human canines. The specimens were sterilized, inoculated with P. gingivalis (ATCC 33277), and incubated for 1 week to allow for biofilm formation. Infected dentin specimens were exposed for 3 days to the following treatments: antibiotic-free polydioxanone scaffold (PDS, control), PDS + 25 wt% TAP [25 mg of each antibiotic (metronidazole, ciprofloxacin, and minocycline) per mL of the PDS polymer solution], or a saturated TAP-based solution (50 mg of each antibiotic per mL of saline solution). In order to serve as the negative control, infected dentin specimens were left untreated (bacteria only). To determine the antimicrobial efficacy of the TAP-mimic scaffold, a colony-forming unit (CFU) per milliliter (n = 10/group) measurement was performed. Furthermore, additional specimens (n = 2/group) were prepared to qualitatively study biofilm inhibition via scanning electron microscopy (SEM). Statistics were performed, and significance was set at the 5% level. Both the TAP-mimic scaffold and the positive control (TAP solution) led to complete bacterial elimination, differing statistically (p < 0.05) from the negative control group (bacteria only). No statistical differences were observed for CFU per milliliter data between antibiotic-free scaffolds (2.7 log10 CFU/mL) and the negative control (5.9 log10 CFU/mL). The obtained data revealed significant antimicrobial properties of the novel PDS-based TAP-mimic scaffold against an established P. gingivalis-infected dentin biofilm. Collectively, the data suggest that the proposed nanofibrous scaffold might be used as an alternative to the advocated clinical gold standard (i.e., TAP) for intracanal disinfection prior to regenerative endodontics.

Incidence of Clostridium perfringens in commercially produced cured raw meat product mixtures and behavior in cooked products during chilling and refrigerated storage.

PubMed

Taormina, Peter J; Bartholomew, Gene W; Dorsa, Warren J

2003-01-01

A total of 445 whole-muscle and ground or emulsified raw pork, beef, and chicken product mixtures acquired from industry sources were monitored over a 10-month period for vegetative and spore forms of Clostridium perfringens. Black colonies that formed on Shahidi-Ferguson perfringens (SFP) agar after 24 h at 37 degrees C were considered presumptive positive. Samples that were positive after a 15-min heat shock at 75 degrees C were considered presumptive positive for spores. Of 194 cured whole-muscle samples, 1.6% were positive; spores were not detected from those samples. Populations of vegetative cells did not exceed 1.70 log10 CFU/g and averaged 1.56 log10 CFU/g. Of 152 cured ground or emulsified samples, 48.7% were positive, and 5.3% were positive for spores. Populations of vegetative cells did not exceed 2.72 log10 CFU/g and averaged 1.98 log10 CFU/g; spores did not exceed 2.00 log10 CFU/g and averaged 1.56 log10 CFU/g. Raw bologna (70% chicken), chunked ham with emulsion, and whole-muscle ham product mixtures were inoculated with C. perfringens spores (ATCC 12916, ATCC 3624, FD1041, and two product isolates) to ca. 3.0 log10 CFU/g before being subjected either to thermal processes mimicking cooking and chilling regimes determined by in-plant temperature probing or to cooking and extended chilling regimes. Populations of C. perfringens were recovered on SFP from each product at the peak cook temperatures, at 54.4, 26.7, and 7.2 degrees C, and after up to 14 days of storage under vacuum at 4.4 degrees C. In each product, populations remained relatively unchanged during chilling from 54.4 to 7.2 degrees C and declined slightly during refrigerated storage. These findings indicate processed meat products cured with sodium nitrite are not at risk for the growth of C. perfringens during extended chilling and cold storage.

Comparison of the disinfection efficacy of chlorine-based products for inactivation of viral indicators and pathogenic bacteria in produce wash water.

PubMed

Chaidez, Cristobal; Moreno, Maria; Rubio, Werner; Angulo, Miguel; Valdez, Benigno

2003-09-01

Outbreaks of pathogenic bacteria infections associated with the consumption of fresh produce has occurred with increased frequency in recent years. This study was undertaken to determine the efficacy of three commonly used disinfectants in packing-houses of Culiacan, Mexico (sodium hypochlorite [NaOCl], trichlor-s-triazinetrione [TST] and thrichlormelamine [TCM]) for inactivation of viral indicators and pathogenic bacteria inoculated onto produce wash water. Each microbial challenge consisted of 2 L of water containing approximately 8 log10 bacterial CFU ml(-1), and 8 log10 viral PFU ml(-1) treated with 100 and 300 mg l(-1) of total chlorine with modified turbidity. Water samples were taken after 2 min of contact with chlorine-based products and assayed for the particular microorganisms. TST and NaOCl were found to effectively reduce for bacterial pathogens and viral indicators 8 log10 and 7 log10, respectively (alpha=0.05). The highest inactivation rate was observed when the turbidity was low and the disinfectant was applied at 300 mg l(-1). TCM did not show effective results when compared with the TST and NaOCl (P<0.05). These findings suggest that turbidity created by the organic and inorganic material present in the water tanks carried by the fresh produce may affect the efficacy of the chlorine-based products.

Control of Salmonella enterica and Listeria monocytogenes in hummus using allyl isothiocyanate.

PubMed

Olaimat, Amin N; Al-Holy, Murad A; Abu Ghoush, Mahmoud; Al-Nabulsi, Anas A; Holley, Richard A

2018-08-02

Hummus (chickpea dip) is a ready-to-eat product which has been implicated in several foodborne outbreaks and food recalls. This study aimed to screen the antimicrobial activity of allyl isothiocyanate (AITC) against 5 strains of each of Salmonella enterica and Listeria monocytogenes using a disc diffusion method. Additionally, the antimicrobial activity of 0.1-1.5% (v/w) AITC against both pathogens and aerobic bacteria in hummus was also investigated. The inhibition zones of AITC were 8.5-15 and 7.0-8.5 mm against the S. enterica and L. monocytogenes strains, respectively, at 37 °C. S. enterica numbers were reduced by >6 log 10 CFU/g in hummus containing ≥0.5% AITC by 3 days at both 4 and 10 °C. While 0.1-0.25% AITC reduced S. enterica by 2.5-5.1 log 10 CFU/g at 4 °C or by 4.7-6.0 log 10 CFU/g at 10 °C by 10 days. Similarly, L. monocytogenes numbers decreased by >6 log 10 CFU/g in hummus with ≥0.5% or ≥1.0% AITC at 4 or 10 °C, respectively, by 3 days. Further, 0.25% AITC significantly reduced L. monocytogenes in hummus by 2.7 and 4.3 log 10 CFU/g at 4 and 10 °C, respectively. Moreover, 0.1% AITC reduced L. monocytogenes by 1.8 log 10 CFU/g in hummus at 10 °C and inhibited the growth at 4 °C for up to 10 days. The aerobic bacterial count also significantly decreased in un-inoculated hummus treated with 1.0-1.5% AITC at both 4 and 10 °C, while a concentration of 0.25-0.5% AITC inhibited their growth at 4 °C. AITC can be used to reduce the risk of salmonellosis or listeriosis in hummus and extend its shelf-life. Copyright © 2018 Elsevier B.V. All rights reserved.

Tuberculous Pericarditis is Multibacillary and Bacterial Burden Drives High Mortality

PubMed Central

Pasipanodya, Jotam G.; Mubanga, Mwenya; Ntsekhe, Mpiko; Pandie, Shaheen; Magazi, Beki T.; Gumedze, Freedom; Myer, Landon; Gumbo, Tawanda; Mayosi, Bongani M.

2015-01-01

Background Tuberculous pericarditis is considered to be a paucibacillary process; the large pericardial fluid accumulation is attributed to an inflammatory response to tuberculoproteins. Mortality rates are high. We investigated the role of clinical and microbial factors predictive of tuberculous pericarditis mortality using the artificial intelligence algorithm termed classification and regression tree (CART) analysis. Methods Patients were prospectively enrolled and followed in the Investigation of the Management of Pericarditis (IMPI) registry. Clinical and laboratory data of 70 patients with confirmed tuberculous pericarditis, including time-to-positive (TTP) cultures from pericardial fluid, were extracted and analyzed for mortality outcomes using CART. TTP was translated to log10 colony forming units (CFUs) per mL, and compared to that obtained from sputum in some of our patients. Findings Seventy patients with proven tuberculous pericarditis were enrolled. The median patient age was 35 (range: 20–71) years. The median, follow up was for 11.97 (range: 0·03–74.73) months. The median TTP for pericardial fluid cultures was 22 (range: 4–58) days or 3.91(range: 0·5–8·96) log10CFU/mL, which overlapped with the range of 3.24–7.42 log10CFU/mL encountered in sputum, a multi-bacillary disease. The overall mortality rate was 1.43 per 100 person-months. CART identified follow-up duration of 5·23 months on directly observed therapy, a CD4 + count of ≤ 199.5/mL, and TTP ≤ 14 days (bacillary load ≥ 5.53 log10 CFU/mL) as predictive of mortality. TTP interacted with follow-up duration in a non-linear fashion. Interpretation Patients with culture confirmed tuberculous pericarditis have a high bacillary burden, and this bacterial burden drives mortality. Thus proven tuberculosis pericarditis is not a paucibacillary disease. Moreover, the severe immunosuppression suggests limited inflammation. There is a need for the design of a highly bactericidal regimen for this condition. PMID:26870789

Production of Gouda cheese and Camembert with probiotic cultures: the suitability of some commercial probiotic cultures to be implemented in cheese.

PubMed

Van de Casteele, S; Ruyssen, T; Vanheuverzwijn, T; Van Assche, P

2003-01-01

The behaviour of 10 probiotic cultures (L. acidophilus, Bifidobacterium sp., L. rhamnosus and L. paracasei) was examined during the production and ripening of Gouda cheese and Camembert. The overall objective of this research project was to obtain a product (cheese) containing at least 10(7) probiotic cfu/g. In general 10(6) cfu of a probiotic culture must be implemented per ml cheese milk, together with the cheesestarter, to reach this objective. L. paracasei sp. have the ability to grow more than 2 log units during cheese ripening. A lower inoculation value can be considered for these cultures.

Integration of an innovative biological treatment with physical or chemical disinfection for wastewater reuse.

PubMed

De Sanctis, Marco; Del Moro, Guido; Levantesi, Caterina; Luprano, Maria Laura; Di Iaconi, Claudio

2016-02-01

In the present paper, the effectiveness of a Sequencing Batch Biofilter Granular Reactor (SBBGR) and its integration with different disinfection strategies (UV irradiation, peracetic acid) for producing an effluent suitable for agricultural use was evaluated. The plant treated raw domestic sewage, and its performances were evaluated in terms of the removal efficiency of a wide group of physical, chemical and microbiological parameters. The SBBGR resulted really efficient in removing suspended solids, COD and nitrogen with an average effluent concentration of 5, 32 and 10 mg/L, respectively. Lower removal efficiency was observed for phosphorus with an average concentration in the effluent of 3 mg/L. Plant effluent was also characterized by an average electrical conductivity and sodium adsorption ratio of 680 μS/cm and 2.9, respectively. Therefore, according to these gross parameters, the SBBGR effluent was conformed to the national standards required in Italy for agricultural reuse. Moreover, disinfection performances of the SBBGR was higher than that of conventional municipal wastewater treatment plants and met the quality criteria suggested by WHO (Escherichia coli<1000 CFU/100 mL) for agricultural reuse. In particular, the biological treatment by SBBGR removed 3.8±0.4 log units of Giardia lamblia, 2.8±0.8 log units of E. coli, 2.5±0.7 log units of total coliforms, 2.0±0.3 log units of Clostridium perfringens, 2.0±0.4 log units of Cryptosporidium parvum and 1.7±0.7 log units of Somatic coliphages. The investigated disinfection processes (UV and peracetic acid) resulted very effective for total coliforms, E. coli and somatic coliphages. In particular, a UV radiation and peracetic acid doses of 40 mJ/cm(2) and 1 mg/L respectively reduced E. coli content in the effluent below the limit for agricultural reuse in Italy (10 CFU/100 mL). Conversely, they were both ineffective on C.perfringens spores. Copyright © 2015 Elsevier B.V. All rights reserved.

In vitro characterization of PlySK1249, a novel phage lysin, and assessment of its antibacterial activity in a mouse model of Streptococcus agalactiae bacteremia.

PubMed

Oechslin, Frank; Daraspe, Jean; Giddey, Marlyse; Moreillon, Philippe; Resch, Grégory

2013-12-01

Beta-hemolytic Streptococcus agalactiae is the leading cause of bacteremia and invasive infections. These diseases are treated with β-lactams or macrolides, but the emergence of less susceptible and even fully resistant strains is a cause for concern. New bacteriophage lysins could be promising alternatives against such organisms. They hydrolyze the bacterial peptidoglycan at the end of the phage cycle, in order to release the phage progeny. By using a bioinformatic approach to screen several beta-hemolytic streptococci, a gene coding for a lysin was identified on a prophage carried by Streptococcus dysgalactiae subsp. equisimilis SK1249. The gene product, named PlySK1249, harbored an original three-domain structure with a central cell wall-binding domain surrounded by an N-terminal amidase and a C-terminal CHAP domain. Purified PlySK1249 was highly lytic and bactericidal for S. dysgalactiae (2-log10 CFU/ml decrease within 15 min). Moreover, it also efficiently killed S. agalactiae (1.5-log10 CFU/ml decrease within 15 min) but not several streptococcal commensal species. We further investigated the activity of PlySK1249 in a mouse model of S. agalactiae bacteremia. Eighty percent of the animals (n = 10) challenged intraperitoneally with 10(6) CFU of S. agalactiae died within 72 h, whereas repeated injections of PlySK1249 (45 mg/kg 3 times within 24 h) significantly protected the mice (P < 0.01). Thus, PlySK1249, which was isolated from S. dysgalactiae, demonstrated high cross-lytic activity against S. agalactiae both in vitro and in vivo. These encouraging results indicated that PlySK1249 might represent a good candidate to be developed as a new enzybiotic for the treatment of systemic S. agalactiae infections.

Persistence of Escherichia coli O157:H7 during pilot-scale processing of iceberg lettuce using flume water containing peroxyacetic acid-based sanitizers and various organic loads.

PubMed

Davidson, Gordon R; Kaminski-Davidson, Chelsea N; Ryser, Elliot T

2017-05-02

In order to minimize cross-contamination during leafy green processing, chemical sanitizers are routinely added to the wash water. This study assessed the efficacy of peroxyacetic acid and mixed peracid against E. coli O157:H7 on iceberg lettuce, in wash water, and on equipment during simulated commercial production in a pilot-scale processing line using flume water containing various organic loads. Iceberg lettuce (5.4kg) inoculated to contain 10 6 CFU/g of a 4-strain cocktail of non-toxigenic, GFP-labeled, ampicillin-resistant E. coli O157:H7, was shredded using a commercial shredder, step-conveyed to a flume tank, washed for 90s using water alone or two different sanitizing treatments (50ppm peroxyacetic acid or mixed peracid) in water containing organic loads of 0, 2.5, 5 or 10% (w/v) blended iceberg lettuce, and then dried using a shaker table and centrifugal dryer. Thereafter, three 5.4-kg batches of uninoculated iceberg lettuce were identically processed. Various product (25g) and water (50ml) samples collected during processing along with equipment surface samples (100cm 2 ) from the flume tank, shaker table and centrifugal dryer were then assessed for numbers of E. coli O157:H7. Organic load rarely impacted (P>0.05) the efficacy of either peroxyacetic acid or mixed peracid, with typical reductions of >5logCFU/ml in wash water throughout processing for all organic loads. Increases in organic load in the wash water corresponded to changes in total solids, chemical oxygen demand, turbidity, maximum filterable volume, and oxidation/reduction potential. After 90s of exposure to flume water, E. coli O157:H7 reductions on inoculated lettuce ranged from 0.97 to 1.74logCFU/g using peroxyacetic acid, with an average reduction of 1.35logCFU/g for mixed peracid. E. coli O157:H7 persisted on all previously uninoculated lettuce following the inoculated batch, emphasizing the need for improved intervention strategies that can better ensure end-product safety. Copyright © 2017 Elsevier B.V. All rights reserved.

Addition to thermized milk of Lactococcus lactis subsp. cremoris M104, a wild, novel nisin a-producing strain, replaces the natural antilisterial activity of the autochthonous raw milk microbiota reduced by thermization.

PubMed

Lianou, Alexandra; Samelis, John

2014-08-01

Recent research has shown that mild milk thermization treatments routinely used in traditional Greek cheese production are efficient to inactivate Listeria monocytogenes and other pathogenic or undesirable bacteria, but they also inactivate a great part of the autochthonous antagonistic microbiota of raw milk. Therefore, in this study, the antilisterial activity of raw or thermized (63°C, 30 s) milk in the presence or absence of Lactococcus lactis subsp. cremoris M104, a wild, novel, nisin A-producing (Nis-A+) raw milk isolate, was assessed. Bulk milk samples were taken from a local cheese plant before or after thermization and were inoculated with a five-strain cocktail of L. monocytogenes (approximately 4 log CFU/ml) or with the cocktail, as above, plus the Nis-A+ strain (approximately 6 log CFU/ml) as a bioprotective culture. Heat-sterilized (121°C, 5 min) raw milk inoculated with L. monocytogenes was used as a control treatment. All milk samples were incubated at 37°C for 6 h and then at 18°C for an additional 66 h. L. monocytogenes grew abundantly (>8 log CFU/ml) in heat-sterilized milk, whereas its growth was completely inhibited in all raw milk samples. Conversely, in thermized milk, L. monocytogenes increased by 2 log CFU/ml in the absence of strain M104, whereas its growth was completely inhibited in the presence of strain M104. Furthermore, nisin activity was detected only in milk samples inoculated with strain M104. Thus, postthermal supplementation of thermized bulk milk with bioprotective L. lactis subsp. cremoris cultures replaces the natural antilisterial activity of raw milk reduced by thermization.

Prevalence of pathogenic bacteria in street vended ready-to-eat meats in Windhoek, Namibia.

PubMed

Shiningeni, Daphney; Chimwamurombe, Percy; Shilangale, Renatus; Misihairabgwi, Jane

2018-05-31

To determine the prevalence of pathogenic bacteria in street vended ready-to-eat meats in Windhoek, Namibia, a total of 96 street vended ready to eat meat samples were evaluated. Prevalences of 42%, 52%, 15%, 6% and 83% were observed for Escherichia coli, Staphylococcus aureus, Listeria monocytogenes, Shigella and Enterobacteriaceae respectively, while the highest aerobic plate counts were 7.74 Log 10 cfu/g, 5.67 Log 10 cfu/g, 5.12 Log 10 cfu/g , 4.56 Log 10 cfu/g, 3.3 Log 10 cfu/g, 5.75 Log 10 cfu/g respectively. Unsatisfactory microbial levels were 32% for aerobic plate count, 26% for Enterobacteriaceae, 35% for Escherichia coli, 11% for Listeria monocytogenes, 7% for Staphylococcus aureus and 6% for Shigella. Salmonella was detected in 11% and 40% of samples from two suburbs. The unsatisfactory microbiological quality of some ready-to-eat meats necessitates the provision of training on food safety and hygiene to street vendors for consumer protection. Copyright © 2018 Elsevier Ltd. All rights reserved.

Effect of gamma irradiation on microbial quality of minimally processed carrot and lettuce: A case study in Greater Accra region of Ghana

NASA Astrophysics Data System (ADS)

Frimpong, G. K.; Kottoh, I. D.; Ofosu, D. O.; Larbi, D.

2015-05-01

The effect of ionizing radiation on the microbiological quality on minimally processed carrot and lettuce was studied. The aim was to investigate the effect of irradiation as a sanitizing agent on the bacteriological quality of some raw eaten salad vegetables obtained from retailers in Accra, Ghana. Minimally processed carrot and lettuce were analysed for total viable count, total coliform count and pathogenic organisms. The samples collected were treated and analysed for a 15 day period. The total viable count for carrot ranged from 1.49 to 14.01 log10 cfu/10 g while that of lettuce was 0.70 to 8.5 7 log10 cfu/10 g. It was also observed that total coliform count for carrot was 1.46-7.53 log10 cfu/10 g and 0.14-7.35 log10 cfu/10 g for lettuce. The predominant pathogenic organisms identified were Bacillus cereus, Cronobacter sakazakii, Staphylococcus aureus, and Klebsiella spp. It was concluded that 2 kGy was most effective for medium dose treatment of minimally processed carrot and lettuce.

Industrial Cooling Tower Disinfection Treatment to Prevent Legionella spp.

PubMed Central

Iervolino, Matteo; Mancini, Benedetta; Cristino, Sandra

2017-01-01

The contamination of industrial cooling towers has been identified as one cause of legionellosis, but the real risk has been underestimated. Two different disinfection treatments were tested on Legionella colonization in an industrial Cooling Tower System (CTS). Environmental monitoring of Legionella, P. aeruginosa, and a heterotrophic plate count (HPC) at 36 °C was performed from June to October 2016. The disinfection procedures adopted were based on hydrogen peroxide (H2O2) and silver salts (Ag+), in addition to an anti-algal treatment, then using hyperclorination as a shock, and then continuous treatment by sodium hypochlorite (NaClO). L. pneumophila serogroup 8 was found at a concentration of 5.06 Log cfu/L after the CTS filling; a shock treatment performed by H2O2/Ag+ produced a rapid increase in contamination up to 6.14 Log cfu/L. The CTS activity was stopped and two subsequent shock treatments were performed using NaClO, followed by continuous hyperclorination. These procedures showed a significant decrease (p < 0.05) in Legionella concentration (1.77 Log cfu/L). The same trend was observed for P. aeruginosa (0.55 Log cfu/100 mL) and HPC (1.95 Log cfu/mL) at 36 °C. Environmental monitoring and the adoption of maintenance procedures, including anti-scale treatment, and physical, chemical, and microbiological control, ensure the good performance of a CTS, reducing the Legionella risk for public health. PMID:28954435

Evaluation of the effects of ultraviolet light on bacterial contaminants inoculated into whole milk and colostrum, and on colostrum immunoglobulin G

PubMed Central

Pereira, R. V.; Bicalho, M. L.; Machado, V. S.; Lima, S.; Teixeira, A. G.; Warnick, L. D.; Bicalho, R. C.

2015-01-01

Raw milk and colostrum can harbor dangerous micro-organisms that can pose serious health risks for animals and humans. According to the USDA, more than 58% of calves in the United States are fed unpasteurized milk. The aim of this study was to evaluate the effect of UV light on reduction of bacteria in milk and colostrum, and on colostrum IgG. A pilot-scale UV light continuous (UVC) flow-through unit (45 J/cm2) was used to treat milk and colostrum. Colostrum and sterile whole milk were inoculated with Listeria innocua, Mycobacterium smegmatis, Salmonella serovar Typhimurium, Escherichia coli, Staphylococcus aureus, Streptococcus agalactiae, and Acinetobacter baumannii before being treated with UVC. During UVC treatment, samples were collected at 5 time points and bacteria were enumerated using selective media. The effect of UVC on IgG was evaluated using raw colostrum from a nearby dairy farm without the addition of bacteria. For each colostrum batch, samples were collected at several different time points and IgG was measured using ELISA. The UVC treatment of milk resulted in a significant final count (log cfu/mL) reduction of Listeria monocytogenes (3.2 ± 0.3 log cfu/mL reduction), Salmonella spp. (3.7 ± 0.2 log cfu/mL reduction), Escherichia coli (2.8 ± 0.2 log cfu/mL reduction), Staph. aureus (3.4 ± 0.3 log cfu/mL reduction), Streptococcus spp. (3.4 ± 0.4 log cfu/mL reduction), and A. baumannii (2.8 ± 0.2 log cfu/mL reduction). The UVC treatment of milk did not result in a significant final count (log cfu/mL) reduction for M. smegmatis (1.8 ± 0.5 log cfu/mL reduction). The UVC treatment of colostrum was significantly associated with a final reduction of bacterial count (log cfu/mL) of Listeria spp. (1.4 ± 0.3 log cfu/mL reduction), Salmonella spp. (1.0 ± 0.2 log cfu/mL reduction), and Acinetobacter spp. (1.1 ± 0.3 log cfu/mL reduction), but not of E. coli (0.5 ± 0.3 log cfu/mL reduction), Strep. agalactiae (0.8 ± 0.2 log cfu/mL reduction), and Staph. aureus (0.4 ± 0.2 log cfu/mL reduction). The UVC treatment of colostrum significantly decreased the IgG concentration, with an observed final mean IgG reduction of approximately 50%. Development of new methods to reduce bacterial contaminants in colostrum must take into consideration the barriers imposed by its opacity and organic components, and account for the incidental damage to IgG caused by manipulating colostrum. PMID:24582452

Quantification of Campylobacter jejuni contamination on chicken carcasses in France.

PubMed

Duqué, Benjamin; Daviaud, Samuel; Guillou, Sandrine; Haddad, Nabila; Membré, Jeanne-Marie

2018-04-01

Highly prevalent in poultry, Campylobacter is a foodborne pathogen which remains the primary cause of enteritis in humans. Several studies have determined prevalence and contamination level of this pathogen throughout the food chain. However it is generally performed in a deterministic way without considering heterogeneity of contamination level. The purpose of this study was to quantify, using probabilistic tools, the contamination level of Campylobacter spp. on chicken carcasses after air-chilling step in several slaughterhouses in France. From a dataset (530 data) containing censored data (concentration <10CFU/g), several factors were considered, including the month of sampling, the farming method (standard vs certified) and the sampling area (neck vs leg). All probabilistic analyses were performed in R using fitdistrplus, mc2d and nada packages. The uncertainty (i.e. error) generated by the presence of censored data was small (ca 1 log 10 ) in comparison to the variability (i.e. heterogeneity) of contamination level (3 log 10 or more), strengthening the probabilistic analysis and facilitating result interpretation. The sampling period and sampling area (neck/leg) had a significant effect on Campylobacter contamination level. More precisely, two "seasons" were distinguished: one from January to May, another one from June to December. During the June-to-December season, the mean Campylobacter concentration was estimated to 2.6 [2.4; 2.8] log 10 (CFU/g) and 1.8 [1.5; 2.0] log 10 (CFU/g) for neck and leg, respectively. The probability of having >1000CFU/g (higher limit of European microbial criterion) was estimated to 35.3% and 12.6%, for neck and leg, respectively. In contrast, during January-to-May season, the mean contamination level was estimated to 1.0 [0.6; 1.3] log 10 (CFU/g) and 0.6 [0.3; 0.9] log 10 (CFU/g) for neck and leg, respectively. The probability of having >1000CFU/g was estimated to 13.5% and 2.0% for neck and leg, respectively. An accurate quantification of contamination level enables industrials to better adapt their processing and hygiene practices. These results will also help in refining exposure assessment models. Copyright © 2017 Elsevier Ltd. All rights reserved.

Multispecies Biofilm Development on Space Station Heat Exhanger Core Material

NASA Technical Reports Server (NTRS)

Pyle, B. H.; Roth, S. R.; Vega, L. M.; Pickering, K. D.; Alvarez, Pedro J. J.; Roman, M. C.

2007-01-01

Investigations of microbial contamination of the cooling system aboard the International Space Station (ISS) suggested that there may be a relationship between heat exchanger (HX) materials and the degree of microbial colonization and biofilm formation. Experiments were undertaken to test the hypothesis that biofilm formation is influenced by the type and previous exposure of HX surfaces. Acidovorax delafieldii, Comamonas acidovorans, Hydrogenophaga pseudoflava, Pseudomonas stutzeri, Sphingomonas paucimobilis, and Stenotrophomonas maltophilia, originally isolated from ISS cooling system fluid, were cultured on R2A agar and suspended separately in fresh filter-sterilized ISS cooling fluid, pH 8.3. Initial numbers in each suspension ranged from 10(exp 6)-10(exp 7) CFU/ml, and a mixture contained greater than 10(exp 7) CFU/ml. Coupons of ISS HX material, previously used on orbit (HXOO) or unused (HXUU), polycarbonate (PC) and 316L polished stainless steel (SS) were autoclaved, covered with multispecies suspension in sterile tubes and incubated in the dark at ambient (22-25 C). Original HX material contained greater than 90% Ni, 4.5% Si, and 3.2% B, with a borate buffer. For approximately 10 weeks, samples of fluid were plated on R2A agar, and surface colonization assessed by SYBR green or BacLight staining and microscopy. Suspension counts for the PC and SC samples remained steady at around 10(exp 7) CFU/ml. HXUU counts declined about 1 log in 21 d then remained steady, and HXOO counts declined 2 logs in 28 d, fluctuated and stabilized about 10(exp 3) CFU/ml from 47-54 d. Predominantly yellow S. paucimobilis predominated on plates from HXOO samples up to 26 d, then white or translucent colonies of other species appeared. All colony types were seen on plates from other samples throughout the trial. Epifluorescence microscopy indicated microbial growth on all surfaces by 21 d, followed by variable colonization. After 54 d, all but the HXOO samples had well-distributed live and dead cells; the HXOO samples had few cells and most were live by BacLight. The results suggest that HX materials themselves are inhibiting microbial growth on the surfaces. The HX exposed on orbit to cooling system fluid inhibited growth of some species originally isolated from the system, whereas the unused HX material had a moderate effect compared to no inhibition with PC or SS controls. It is possible that chemistry or microbiology of the ISS system increased deposition of inhibitory compounds on the HXOO coupon surfaces; these may inhibit inoculated species to differing degrees.

Trisodium phosphate and sodium hypochlorite are more effective as antimicrobials against Campylobacter and Salmonella on duck as compared to chicken meat.

PubMed

Sarjit, Amreeta; Dykes, Gary A

2015-06-16

Little work has been reported on the use of commercial antimicrobials against foodborne pathogens on duck meat. We investigated the effectiveness of trisodium phosphate (TSP) and sodium hypochlorite (SH) as antimicrobial treatments against Campylobacter and Salmonella on duck meat under simulated commercial water chilling conditions. The results were compared to the same treatments on well-studied chicken meat. A six strain Campylobacter or Salmonella cocktail was inoculated (5 ml) at two dilution levels (10(4) and 10(8) cfu/ml) onto 25 g duck or chicken meat with skin and allowed to attach for 10 min. The meat was exposed to three concentrations of pH adjusted TSP (8, 10 and 12% (w/v), pH 11.5) or SH (40, 50 and 60 ppm, pH 5.5) in 30 ml water under simulated spin chiller conditions (4 °C, agitation) for 10 min. In a parallel experiment the meat was placed in the antimicrobial treatments before inoculation and bacterial cocktails were added to the meat after the antimicrobial solution was removed while all other parameters were maintained. Untreated controls and controls using water were included in all experiments. Bacterial numbers were determined on Campylobacter blood-free selective agar and Mueller Hinton agar or xylose deoxycholate agar and tryptone soya agar using the thin agar layer method for Campylobacter and Salmonella, respectively. All TSP concentrations significantly (p<0.05) reduced numbers of Campylobacter (~1.2-6.4 log cfu/cm(2)) and Salmonella (~0.4-6.6 log cfu/cm(2)) on both duck and chicken meat. On duck meat, numbers of Campylobacter were less than the limit of detection at higher concentrations of TSP and numbers of Salmonella were less than the limit of detection at all concentrations of TSP except one. On chicken meat, numbers of Campylobacter and Salmonella were less than the limit of detection only at the lower inoculum level and higher TSP concentrations. By contrast only some of the concentrations of SH significantly (p<0.05) reduced numbers of Campylobacter and Salmonella (~0.2-1.5 log cfu/cm(2)) on both duck and chicken meats. None of the SH treatments resulted in numbers of either pathogen being less than limit of detection. Results indicate that chicken meat has the ability to effectively protect Campylobacter and Salmonella against the impact of trisodium phosphate and sodium hypochlorite while duck meat does not. This study suggests that trisodium phosphate has a strong potential for application in a commercial poultry processing to reduce Campylobacter and Salmonella specifically on duck meat. Copyright © 2015 Elsevier B.V. All rights reserved.

Microbiological survey of a South African poultry processing plant.

PubMed

Geornaras, I; de Jesus, A; van Zyl, E; von Holy, A

1995-01-01

Bacterial populations associated with poultry processing were determined on neck skin samples, equipment surfaces and environmental samples by replicate surveys. Aerobic plate counts, Enterobacteriaceae counts, Enterobacteriaceae counts and Pseudomonas counts were performed by standard procedures and the prevalence of Listeria, presumptive Salmonella and Staphylococcus aureus determined. Statistically significant (P < 0.05) increases in counts of all types of bacteria were obtained on product samples as a result of processing. Although bacterial counts on neck skin samples decreased by 0.3 to 0.4 log CFU g-1 after spray washing of carcasses, subsequent spinchilling and packaging of whole carcasses resulted in 0.7 to 1.2 log CFU g-1 increases. Bacterial numbers on equipment surfaces, however, decreased significantly from the "dirty" to the "clean" areas of the abattoir. Transport cages, "rubber fingers", defeathering curtains, shackles and conveyor belts repeatedly showed aerobic plate counts in excess of 5.0 log CFU 25 cm-2. Aerobic plate counts of scald tank and spinchiller water were 2 log CFU ml-1 higher than those of potable water samples. Bacterial numbers of the air in the "dirty" area were higher than those of the "clean" area. Listeria, presumptive Salmonella and Staphylococcus aureus were isolated from 27.6, 51.7 and 24.1% of all product samples, respectively, and Listeria and Staphylococcus aureus were also isolated from selected equipment surfaces.

Adhesion of Stenotrophomonas maltophilia, Delftia acidovorans, and Achromobacter xylosoxidans to Contact Lenses.

PubMed

Vijay, Ajay Kumar; Willcox, Mark D P

2017-09-26

Contact lens cases become contaminated with microbes during use. We wished to compare the adhesion of uncommon bacterial contaminants isolated from lens cases to contact lenses with and without organic soil. Strains of Delftia acidovorans (001), Stenotrophomonas maltophilia (002 and 006), and Achromobacter xylosoxidans (001) isolated from contact lens cases (test strains) and Pseudomonas aeruginosa (Paer1) isolated from eyes at the time of infiltrative response (control strain) were used. Bacteria were grown and resuspended in phosphate-buffered saline (PBS) or 10% organic soil (heat-killed Saccharomyces cerevisiae resuspended in complement inactivated bovine serum). Two silicone hydrogel (senofilcon A and comfilcon A) and one hydrogel lens (etafilcon A) lens materials were used. Bacteria (1.0×10 and 1.0×10 colony-forming units/mL; CFU/mL) adhered to lenses for 24 hr and the numbers of bacteria adherent to each lens type (with and without organic soil) were estimated by culture. All the four test strains adhered in significantly greater numbers to contact lenses after incubation in inoculum prepared with organic soil compared with PBS-D. acidovorans 001 (0.7 log10 CFU; P<0.05), S. maltophilia 002 (1.7 log10 CFU; P<0.05), S. maltophilia 006 (0.9 log10 CFU; P<0.05), and A. xylosoxidans 001 (0.4 log10 CFU; P<0.05). However, the presence of organic soil did not increase adhesion of P. aeruginosa Paer1 (-0.1 log10 CFU; P>0.05). Achromobacter xylosoxidans 001 (P<0.01), D. acidovorans 001 (P<0.01), and S. maltophilia 002 (P<0.01) significantly differed in their adhesion to the three contact lens materials. Bacteria that are commonly found in contact lens cases adhered to contact lenses in relatively high numbers in the presence of organic soil. This might indicate that a similar phenomenon occurs in the presence of tears. This may facilitate their transfer from the lens to the cornea and the production of corneal infiltrates.

In Vitro Studies of Pharmacodynamic Properties of Vancomycin against Staphylococcus aureus and Staphylococcus epidermidis

PubMed Central

Löwdin, E.; Odenholt, I.; Cars, O.

1998-01-01

The bactericidal activities of vancomycin against two reference strains and two clinical isolates of Staphylococcus aureus and Staphylococcus epidermidis were studied with five different concentrations ranging from 2× to 64× the MIC. The decrease in the numbers of CFU at 24 h was at least 3 log10 CFU/ml for all strains. No concentration-dependent killing was observed. The postantibiotic effect (PAE) was determined by obtaining viable counts for two of the reference strains, and the viable counts varied markedly: 1.2 h for S. aureus and 6.0 h for S. epidermidis. The determinations of the PAE, the postantibiotic sub-MIC effect (PA SME), and the sub-MIC effect (SME) for all strains were done with BioScreen C, a computerized incubator for bacteria. The PA SMEs were longer than the SMEs for all strains tested. A newly developed in vitro kinetic model was used to expose the bacteria to continuously decreasing concentrations of vancomycin. A filter prevented the loss of bacteria during the experiments. One reference strain each of S. aureus and S. epidermidis and two clinical isolates of S. aureus were exposed to an initial concentration of 10× the MIC of vancomycin with two different half-lives (t1/2s): 1 or 5 h. The post-MIC effect (PME) was calculated as the difference in time for the bacteria to grow 1 log10 CFU/ml from the numbers of CFU obtained at the time when the MIC was reached and the corresponding time for an unexposed control culture. The difference in PME between the strains was not as pronounced as that for the PAE. Furthermore, the PME was shorter when a t1/2 of 5 h (approximate terminal t1/2 in humans) was used. The PMEs at t1/2s of 1 and 5 h were 6.5 and 3.6 h, respectively, for S. aureus. The corresponding figures for S. epidermidis were 10.3 and less than 6 h. The shorter PMEs achieved with a t1/2 of 5 h and the lack of concentration-dependent killing indicate that the time above the MIC is the parameter most important for the efficacy of vancomycin. PMID:9756787

Release of antibiotic resistant bacteria and genes in the effluent and biosolids of five wastewater utilities in Michigan.

PubMed

Munir, Mariya; Wong, Kelvin; Xagoraraki, Irene

2011-01-01

The purpose of this study was to quantify the occurrence and release of antibiotic resistant genes (ARGs) and antibiotic resistant bacteria (ARB) into the environment through the effluent and biosolids of different wastewater treatment utilities including an MBR (Membrane Biological Reactor) utility, conventional utilities (Activated Sludge, Oxidative Ditch and Rotatory Biological Contactors-RBCs) and multiple sludge treatment processes (Dewatering, Gravity Thickening, Anaerobic Digestion and Lime Stabilization). Samples of raw wastewater, pre- and post-disinfected effluents, and biosolids were monitored for tetracycline resistant genes (tetW and tetO) and sulfonamide resistant gene (Sul-I) and tetracycline and sulfonamide resistant bacteria. ARGs and ARB concentrations in the final effluent were found to be in the range of ND(non-detectable)-2.33 × 10(6) copies/100 mL and 5.00 × 10(2)-6.10 × 10(5) CFU/100 mL respectively. Concentrations of ARGs (tetW and tetO) and 16s rRNA gene in the MBR effluent were observed to be 1-3 log less, compared to conventional treatment utilities. Significantly higher removals of ARGs and ARB were observed in the MBR facility (range of removal: 2.57-7.06 logs) compared to that in conventional treatment plants (range of removal: 2.37-4.56 logs) (p < 0.05). Disinfection (Chlorination and UV) processes did not contribute in significant reduction of ARGs and ARB (p > 0.05). In biosolids, ARGs and ARB concentrations were found to be in the range of 5.61 × 10(6)-4.32 × 10(9) copies/g and 3.17 × 10(4)-1.85 × 10(9) CFU/g, respectively. Significant differences (p < 0.05) were observed in concentrations of ARGs (except tetW) and ARB between the advanced biosolid treatment methods (i.e., anaerobic digestion and lime stabilization) and the conventional dewatering and gravity thickening methods. Copyright © 2010 Elsevier Ltd. All rights reserved.

Evaluation of the efficacy of bleach routinely used in health facilities against Mycobacterium tuberculosis isolates in Ethiopia

PubMed Central

Mekonnen, Daniel; Admassu, Aschalew; Wassie, Belaynew; Biadglegne, Fantahun

2015-01-01

Introduction In Ethiopia, the most widely used disinfectant is 5% Hypochlorites. However, Ethiopian national health safety and infection prevention guideline recommendation on the use of bleach is not consistent and varying from 0.1%-4%. The purpose of this study was therefore to assess the effective time-concentration relationship of sodium hypochlorite against Mycobacterium tuberculosis complex isolates in the absence of any organic load. Methods This experimental study was conducted in Bahir Dar Regional laboratory from February-June 2013. Test suspensions of 1.5 X 108 CFU/ml prepared using normal saline containing 0.5% tween 80. From 5% stock, 0.1%, 0.5%, 1% and 2% bleach was prepared. A 1ml of test strain suspension and 1ml of bleach mixed and allowed to stand until the specified time achieved, neutralized by 48 ml phosphate buffer. 100µl from the diluted sediment were spread on two L-J mediums and incubated at 37°C for 8 weeks. Results When 0. 1% bleach was used for 10 min, majority 11/20 of isolates showed 3 x 103 CFU/ml growth (ME = 4.4) which was inefficient. However, when the time increased, the log10 reduction was acceptable, ME >5 and it was effective. The bleach solution containing 0.5% and above was effective in all respective times. In this study, there is no difference observed in the tuberculocidal activity of bleach against resistant and sensitive strains. Conclusion Our study showed that in the absence of any organic load, 0.1% bleaches over 15 min and 0.5% bleaches over 10 min was found to be tuberculocidal. PMID:26668688

Load and Prevalence of Antimicrobial-Resistant Escherichia coli from Fresh Goat Meat in Arusha, Tanzania.

PubMed

Mwanyika, Gaspary; Call, Douglas R; Rugumisa, Benardether; Luanda, Catherine; Murutu, Rehema; Subbiah, Murugan; Buza, Joram

2016-09-01

Given the potential public health risks associated with a burgeoning goat meat industry in Tanzania, we estimated the load of Escherichia coli and the prevalence of antibiotic-resistant strains for goat meat by using a cross-sectional study design (June to July 2015). Five large (n = 60 samples) and five small (n = 64 samples) slaughterhouses were sampled over a period of four to six visits each. Meat rinsate was prepared and plated onto MacConkey agar, and presumptive E. coli colonies were enumerated and reported as CFU per milliliter of rinsate. In total, 2,736 presumptive E. coli isolates were tested for antibiotic drug sensitivity by using breakpoint assays against 11 medically important antibiotics. E. coli was recovered from almost all the samples (96.8%), with counts ranging from 2 to 4 log CFU ml -1 , and there was no significant difference (P = 0.43) in recovery according to facility size (average, 3.37 versus 3.13 log CFU ml -1 , large and small, respectively). Samples from large facilities had relatively higher prevalence (P = 0.026) of antibiotic-resistant E. coli compared with small facilities. This was mostly explained by more ampicillin (30.1 versus 12.8%) and amoxicillin (17.6 versus 4.5%) resistance for large versus small facilities, respectively, and more tetracycline resistance for small facilities (5.6 versus 10.6%, respectively). Large slaughter operations may serve as foci for dissemination of antibiotic-resistant bacteria via food products. More effective hygiene practices during slaughter and meat handling would limit the probability of transmitting antibiotic-resistant E. coli in goat meat.

Presence of faecal indicator bacteria in groundwaters in Kathmandu Valley, Nepal

NASA Astrophysics Data System (ADS)

Nishida, K.; Shrestha, S.; Tanaka, Y.; Haramoto, E.; Nakamura, T.; Osaka, K.; Chapagain, S.

2010-12-01

Groundwater quality is a critical problem in Kathmandu Valley, Nepal. The population of the city increased 6 times in the last six decades and more than half of water demand depends on groundwater resource. Nevertheless, few data of microorganism presence have been reported qualitatively in the central area of the valley. We investigated distribution of faecal indicator bacteria (Total coliforms and Escherichia coli) detected in wells and analyzed the variations of the concentrations. Groundwater samples were collected from 12 deep tube wells (170-300m depths) and 36 shallow tube wells and dug wells (3-20m depths) in Aug 2008, Jan 2009, Aug 2009 and Aug 2010. River waters were also collected for analyzing effect on groundwater quality. E. coli was detected from most of all wells; the concentrations were within 1 log cfu/100mL in deep tube wells and shallow tube wells while those in dug wells ranged from 1 to 3 log cfu/100mL. E. coli was detected at extremely high level in river water, from 5 to 7 log cfu/100mL, however, no clear relation was observed between E. coli concentrations in any types of groundwaters and distance of wells from adjacent rivers. These results indicate that both types of tube wells were rather protected and dug wells were most vulnerable for faecal contamination at very local scale. Genetic analysis of bacterial communities for deep well samples showed the existence of Enterobacter, Acinetobacter as well as Methane-metabolizing groups which provide information of possible indicators other than total coliforms or E. coli for groundwater management in the valley.

Growth of Escherichia coli O157:H7, Non-O157 Shiga Toxin-Producing Escherichia coli , and Salmonella in Water and Hydroponic Fertilizer Solutions.

PubMed

Shaw, Angela; Helterbran, Kara; Evans, Michael R; Currey, Christopher

2016-12-01

The desire for local, fresh produce year round is driving the growth of hydroponic growing systems in the United States. Many food crops, such as leafy greens and culinary herbs, grown within hydroponics systems have their root systems submerged in recirculating nutrient-dense fertilizer solutions from planting through harvest. If a foodborne pathogen were introduced into this water system, the risk of contamination to the entire crop would be high. Hence, this study was designed to determine whether Escherichia coli O157:H7, non-O157 Shiga toxin-producing E. coli , and Salmonella were able to survive and reproduce in two common hydroponic fertilizer solutions and in water or whether the bacteria would be killed or suppressed by the fertilizer solutions. All the pathogens grew by 1 to 6 log CFU/ml over a 24-h period, depending on the solution. E. coli O157:H7 reached higher levels in the fertilizer solution with plants (3.12 log CFU/ml), whereas non-O157 Shiga toxin-producing E. coli and Salmonella reached higher levels in the fertilizer solution without plants (1.36 to 3.77 log CFU/ml). The foodborne pathogens evaluated here survived for 24 h in the fertilizer solution, and populations grew more rapidly in these solutions than in plain water. Therefore, human pathogens entering the fertilizer solution tanks in hydroponic systems would be expected to rapidly propagate and spread throughout the system and potentially contaminate the entire crop.

Modeling time to inactivation of Listeria monocytogenes in response to high pressure, sodium chloride, and sodium lactate.

PubMed

Youart, Alyssa M; Huang, Yang; Stewart, Cynthia M; Kalinowski, Robin M; Legan, J David

2010-10-01

A mathematical model was developed to predict time to inactivation (TTI) by high pressure processing of Listeria monocytogenes in a broth system (pH 6.3) as a function of pressure (450 to 700 MPa), inoculum level (2 to 6 log CFU/ml), sodium chloride (1 or 2%), and sodium lactate (0 or 2.5%) from a 4°C initial temperature. Ten L. monocytogenes isolates from various sources, including processed meats, were evaluated for pressure resistance. The five most resistant strains were used as a cocktail to determine TTI and for model validation. Complete inactivation of L. monocytogenes in all treatments was demonstrated with an enrichment method. The TTI increased with increasing inoculum level and decreasing pressure magnitude, from 1.5 min at 700 MPa and 2 log CFU/ml, to 15 min at 450 MPa and 6 log CFU/ml. Neither NaCl nor sodium lactate significantly influenced TTI. The model was validated with ready-to-eat, uncured, Australian retail poultry products, and with product specially made at a U.S. Department of Agriculture, Food Safety and Inspection Service (FSIS)-inspected pilot plant in the United States. Data from the 210 individual product samples used for validation indicate that the model gives "fail-safe" predictions (58% with response as expected, 39% with no survivors where survivors expected, and only 3% with survivors where none were expected). This model can help manufacturers of refrigerated ready-to-eat meats establish effective processing criteria for the use of high pressure processing as a postlethality treatment for L. monocytogenes in accordance with FSIS regulations.

Evaluation of the effects of ultraviolet light on bacterial contaminants inoculated into whole milk and colostrum, and on colostrum immunoglobulin G.

PubMed

Pereira, R V; Bicalho, M L; Machado, V S; Lima, S; Teixeira, A G; Warnick, L D; Bicalho, R C

2014-05-01

Raw milk and colostrum can harbor dangerous microorganisms that can pose serious health risks for animals and humans. According to the USDA, more than 58% of calves in the United States are fed unpasteurized milk. The aim of this study was to evaluate the effect of UV light on reduction of bacteria in milk and colostrum, and on colostrum IgG. A pilot-scale UV light continuous (UVC) flow-through unit (45 J/cm(2)) was used to treat milk and colostrum. Colostrum and sterile whole milk were inoculated with Listeria innocua, Mycobacterium smegmatis, Salmonella serovar Typhimurium, Escherichia coli, Staphylococcus aureus, Streptococcus agalactiae, and Acinetobacter baumannii before being treated with UVC. During UVC treatment, samples were collected at 5 time points and bacteria were enumerated using selective media. The effect of UVC on IgG was evaluated using raw colostrum from a nearby dairy farm without the addition of bacteria. For each colostrum batch, samples were collected at several different time points and IgG was measured using ELISA. The UVC treatment of milk resulted in a significant final count (log cfu/mL) reduction of Listeria monocytogenes (3.2 ± 0.3 log cfu/mL reduction), Salmonella spp. (3.7 ± 0.2 log cfu/mL reduction), Escherichia coli (2.8 ± 0.2 log cfu/mL reduction), Staph. aureus (3.4 ± 0.3 log cfu/mL reduction), Streptococcus spp. (3.4 ± 0.4 log cfu/mL reduction), and A. baumannii (2.8 ± 0.2 log cfu/mL reduction). The UVC treatment of milk did not result in a significant final count (log cfu/mL) reduction for M. smegmatis (1.8 ± 0.5 log cfu/mL reduction). The UVC treatment of colostrum was significantly associated with a final reduction of bacterial count (log cfu/mL) of Listeria spp. (1.4 ± 0.3 log cfu/mL reduction), Salmonella spp. (1.0 ± 0.2 log cfu/mL reduction), and Acinetobacter spp. (1.1 ± 0.3 log cfu/mL reduction), but not of E. coli (0.5 ± 0.3 log cfu/mL reduction), Strep. agalactiae (0.8 ± 0.2 log cfu/mL reduction), and Staph. aureus (0.4 ± 0.2 log cfu/mL reduction). The UVC treatment of colostrum significantly decreased the IgG concentration, with an observed final mean IgG reduction of approximately 50%. Development of new methods to reduce bacterial contaminants in colostrum must take into consideration the barriers imposed by its opacity and organic components, and account for the incidental damage to IgG caused by manipulating colostrum. Copyright © 2014 American Dairy Science Association. Published by Elsevier Inc. All rights reserved.

Pulsed UV laser light on Escherichia coli and Saccharomyces cerevisiae suspended in non-alcoholic beer

PubMed Central

Hosseini, SM; Azar-Daryany, MK; Massudi, R; Elikaei, A

2011-01-01

Background The aim of this study was to investigate the effect of pulsed ultra-violet (UV) irradiation on inactivation of beer spoilage microorganisms. UV irradiation is nowadays cost effective enough to compete with traditional biological, physical, and chemical treatment technologies and has become an alternative to such methods. Material and Methods Photoinactivation effects of pulsed UV laser with the wavelengths of 355 and 266 nm, which inactivate typical prokaryotic (Escherichia coli) and eukaryotic (Saccharomyces cerevisiae) microorganisms, were examined with different doses and exposure times. Results A dose of 100 J/cm2 of the 355 nm pulsed UV laser was able to reduce about 1 to 2 log (88.75%) of E.coli with the population of 1.6×108 colony-forming units (CFU/ml), and 97% of 3.2×107, 3×106, 5.5×105, and 9×104 CFU/ml. In the case of 266 nm, more than 99% reduction in E. coli serial dilutions was inactivated, using 10 J/cm2 with exception of 7×104 CFU/ml which was not detected any bacterial growth using 5 J/cm2. In addition, 50, 40, and 20 J/cm2 energy were used successfully to inactivate S. cerevisiae at the populations of 5.4×106, 7×105, 5×104 and 4×103 CFU/ml, respectively. As a result, pulsed UV Laser with 266 nm was strong enough to inactivate a high titer of bacterial and yeast indicator standards suspended in non-alcoholic beer in comparison with 355nm doses. Conclusion Results indicate that pulsed UV technology, in principle, is an attractive alternative to conventional methods for the inactivation of indicator microorganisms and has potential in irradiation of unpasteurized beer. PMID:22347580

Detection of bacteriuria and pyuria by URISCREEN a rapid enzymatic screening test.

PubMed Central

Pezzlo, M T; Amsterdam, D; Anhalt, J P; Lawrence, T; Stratton, N J; Vetter, E A; Peterson, E M; de la Maza, L M

1992-01-01

A multicenter study was performed to evaluate the ability of the URISCREEN (Analytab Products, Plainview, N.Y.), a 2-min catalase tube test, to detect bacteriuria and pyuria. This test was compared with the Chemstrip LN (BioDynamics, Division of Boehringer Mannheim Diagnostics, Indianapolis, Ind.), a 2-min enzyme dipstick test; a semiquantitative plate culture method was used as the reference test for bacteriuria, and the Gram stain or a quantitative chamber count method was used as the reference test for pyuria. Each test was evaluated for its ability to detect probable pathogens at greater than or equal to 10(2) CFU/ml and/or greater than or equal to 1 leukocyte per oil immersion field, as determined by the Gram stain method, or greater than 10 leukocytes per microliter, as determined by the quantitative count method. A total of 1,500 urine specimens were included in this evaluation. There were 298 specimens with greater than or equal 10(2) CFU/ml and 451 specimens with pyuria. Of the 298 specimens with probable pathogens isolated at various colony counts, 219 specimens had colony counts of greater than or equal to 10(5) CFU/ml, 51 specimens had between 10(4) and 10(5) CFU/ml, and 28 specimens had between 10(2) and less than 10(4) CFU/ml. Both the URISCREEN and the Chemstrip LN detected 93% (204 of 219) of the specimens with probable pathogens at greater than or equal to 10(5) CFU/ml. For the specimens with probable pathogens at greater than or equal to 10(2) CFU/ml, the sensitivities of the URISCREEN and the Chemstrip LN were 86% (256 of 298) and 81% (241 of 298), respectively. Of the 451 specimens with pyuria, the URISCREEN detected 88% (398 of 451) and Chemstrip LN detected 78% (350 if 451). There were 204 specimens with both greater than or equal to 10(2) CFU/ml and pyuria; the sensitivities of both methods were 95% (193 of 204) for these specimens. Overall, there were 545 specimens with probable pathogens at greater than or equal to 10(2) CFU/ml and/or pyuria. The URISCREEN detected 85% (461 of 545), and the Chemstrip LN detected 73% (398 of 545). A majority (76%) of the false-negative results obtained with either method were for specimens without leukocytes in the urine. There were 955 specimens with no probable pathogens or leukocytes. Of these, 28% (270 of 955) were found positive by the URISCREEN and 13% (122 of 955) were found positive by the Chemstrip LN. A majority of the false-positive results were probably due, in part, to the detection of enzymes present in both bacterial and somatic cells by each of the test systems. Overall, the URISCREEN is rapid, manual, easy-to-perform enzymatic test that yields findings similar to those yielded by the Chemstrip LN for specimens with both greater than or equal to 10(2) CFU/ml and pyuria or for specimens with greater than or equal to 10(5) CFU/ml and with or without pyuria. However, when the data were analyzed for either probable pathogens at less 10(5) CFU/ml or pyuria, the sensitivity of the URISCREEN was higher (P less than 0.05). PMID:1551986

Transmission of Escherichia coli O157:H7 to internal tissues and its survival on flowering heads of wheat.

PubMed

Martinez, Bismarck; Stratton, Jayne; Bianchini, Andréia; Wegulo, Stephen; Weaver, Glen

2015-03-01

Escherichia coli O157:H7 is a human pathogen that can cause bloody diarrhea, hemorrhagic colitis, and hemolytic uremic syndrome. E. coli O157:H7 illnesses are mainly associated with undercooked beef; however, in recent years, outbreaks have been linked to fresh produce, such as spinach, lettuce, and sprouts. In 2009, flour was implicated as the contamination source in an outbreak involving consumption of raw cookie dough that resulted in 77 illnesses. The objectives of this research were to determine (i) whether E. coli O157:H7 could be translocated into the internal tissues of wheat (Triticum aestivum) seedlings from contaminated seed, soil, or irrigation water and (ii) whether the bacterium could survive on flowering wheat heads. The levels of contamination of kanamycin-resistant E. coli O157:H7 strains in seed, soil, and irrigation water were 6.88 log CFU/g, 6.60 log CFU/g, and 6.76 log CFU/ml, respectively. One hundred plants per treatment were sown in pot trays with 50 g of autoclaved soil or purposely contaminated soil, watered every day with 5 ml of water, and harvested 9 days postinoculation. In a fourth experiment, flowering wheat heads were spray inoculated with water containing 4.19 log CFU/ml E. coli O157:H7 and analyzed for survival after 15 days, near the harvest period. To detect low levels of internalization, enrichment procedures were performed and Biotecon real-time PCR detection assays were used to determine the presence of E. coli O157:H7 in the wheat, using a Roche Applied Science LightCycler 2.0 instrument. The results showed that internalization was possible using contaminated seed, soil, and irrigation water in wheat seedlings, with internalization rates of 2, 5, and 10%, respectively. Even though the rates were low, to our knowledge this is the first study to demonstrate the ability of this strain to reach the phylloplane in wheat. In the head contamination experiment, all samples tested positive, showing the ability of E. coli O157:H7 to survive on the wheat head.

Direct comparison of Xpert MTB/RIF assay with liquid and solid mycobacterial culture for quantification of early bactericidal activity.

PubMed

Kayigire, Xavier A; Friedrich, Sven O; Venter, Amour; Dawson, Rodney; Gillespie, Stephen H; Boeree, Martin J; Heinrich, Norbert; Hoelscher, Michael; Diacon, Andreas H

2013-06-01

The early bactericidal activity of antituberculosis agents is usually determined by measuring the reduction of the sputum mycobacterial load over time on solid agar medium or in liquid culture. This study investigated the value of a quantitative PCR assay for early bactericidal activity determination. Groups of 15 patients were treated with 6 different antituberculosis agents or regimens. Patients collected sputum for 16 h overnight at baseline and at days 7 and 14 after treatment initiation. We determined the sputum bacterial load by CFU counting (log CFU/ml sputum, reported as mean ± standard deviation [SD]), time to culture positivity (TTP, in hours [mean ± SD]) in liquid culture, and Xpert MTB/RIF cycle thresholds (C(T), n [mean ± SD]). The ability to discriminate treatment effects between groups was analyzed with one-way analysis of variance (ANOVA). All measurements showed a decrease in bacterial load from mean baseline (log CFU, 5.72 ± 1.00; TTP, 116.0 ± 47.6; C(T), 19.3 ± 3.88) to day 7 (log CFU, -0.26 ± 1.23, P = 0.2112; TTP, 35.5 ± 59.3, P = 0.0002; C(T), 0.55 ± 3.07, P = 0.6030) and day 14 (log CFU, -0.55 ± 1.24, P = 0.0006; TTP, 54.8 ± 86.8, P < 0.0001; C(T), 2.06 ± 4.37, P = 0.0020). The best discrimination between group effects was found with TTP at day 7 and day 14 (F = 9.012, P < 0.0001, and F = 11.580, P < 0.0001), followed by log CFU (F = 4.135, P = 0.0024, and F = 7.277, P < 0.0001). C(T) was not significantly discriminative (F = 1.995, P = 0.091, and F = 1.203, P = 0.316, respectively). Culture-based methods are superior to PCR for the quantification of early antituberculosis treatment effects in sputum.

Decontamination of materials contaminated with Francisella philomiragia or MS2 bacteriophage using PES-Solid, a solid source of peracetic acid.

PubMed

Buhr, T L; Young, A A; Johnson, C A; Minter, Z A; Wells, C M

2014-08-01

The aim of the study was to develop test methods and evaluate survival of Francisella philomiragia cells and MS2 bacteriophage after exposure to PES-Solid (a solid source of peracetic acid) formulations with or without surfactants. Francisella philomiragia cells (≥7·6 log10 CFU) or MS2 bacteriophage (≥6·8 log10 PFU) were deposited on seven different test materials and treated with three different PES-Solid formulations, three different preneutralized samples and filter controls at room temperature for 15 min. There were 0-1·3 log10 CFU (<20 cells) of cell survival, or 0-1·7 log10 (<51 PFU) of bacteriophage survival in all 21 test combinations (organism, formulation and substrate) containing reactive PES-Solid. In addition, the microemulsion (Dahlgren Surfactant System) showed ≤2 log10 (100 cells) of viable F. philomiragia cells, indicating the microemulsion achieved <2 log10 CFU on its own. Three PES-Solid formulations and one microemulsion system (DSS) inactivated F. philomiragia cells and/or MS2 bacteriophage that were deposited on seven different materials. A test method was developed to show that reactive PES-Solid formulations and a microemulsion system (DSS) inactivated >6 log10 CFU/PFU F. philomiragia cells and/or MS2 bacteriophage on different materials. Published 2014. This article is a U.S. Government work and is in the public domain in the USA.

Inactivation of Salmonella Enteritidis on lettuces used by minimally processed vegetable industries.

PubMed

Silveira, Josete Bailardi; Hessel, Claudia Titze; Tondo, Eduardo Cesar

2017-01-30

Washing and disinfection methods used by minimally processed vegetable industries of Southern Brazil were reproduced in laboratory in order to verify their effectiveness to reduce Salmonella Enteritidis SE86 (SE86) on lettuce. Among the five industries investigated, four carried out washing with potable water followed by disinfection with 200 ppm sodium hypochlorite during different immersion times. The washing procedure alone decreased approximately 1 log CFU/g of SE86 population and immersion times of 1, 2, 5, and 15 minutes in disinfectant solution demonstrated reduction rates ranging from 2.06±0.10 log CFU/g to 3.01±0.21 log CFU/g. Rinsing alone was able to reduce counts from 0.12±0.63 log CFU/g to 1.90±1.07 log CFU/g. The most effective method was washing followed by disinfection with 200 ppm sodium hypochlorite for 15 minutes and final rinse with potable water, reaching 5.83 log CFU/g of reduction. However, no statistical differences were observed on the reduction rates after different immersion times. A time interval of 1 to 2 minutes may be an advantage to the minimally vegetable processed industries in order to optimize the process without putting at risk food safety.

Fungal Presence in Selected Tree Nuts and Dried Fruits

PubMed Central

Tournas, VH; Niazi, NS; Kohn, JS

2015-01-01

Sixty-four tree nut samples (almonds, pecans, pine nuts, and walnuts) and 50 dried fruit samples (apricots, cranberries, papaya, pineapple, and raisins) were purchased from local supermarkets and analyzed for fungal contamination using conventional culture as well as molecular methods. The results of our study showed that the highest yeast and mold (YM) counts (5.34 log10 CFU g−1) were found in walnuts and the lowest in pecans. The most common mold in nuts was Aspergillus niger, relatively low numbers of A. flavus were found across the board, while Penicillium spp. were very common in pine nuts and walnuts. Low levels (2.00–2.84 log10 CFU g−1) of yeasts were recovered from only two pine nut samples. Fungal contamination in dried fruits was minimal (ranging from <2.00 to 3.86 log10 CFU g−1). The highest fungal levels were present in raisins. All papaya samples and the majority of cranberry, pineapple, and apricot samples were free of live fungi. The most common mold in dried fruits was A. niger followed by Penicillium spp. One apricot sample also contained low levels (2.00 log10 CFU g−1) of yeasts. PMID:26056470

Fungal Presence in Selected Tree Nuts and Dried Fruits.

PubMed

Tournas, V H; Niazi, N S; Kohn, J S

2015-01-01

Sixty-four tree nut samples (almonds, pecans, pine nuts, and walnuts) and 50 dried fruit samples (apricots, cranberries, papaya, pineapple, and raisins) were purchased from local supermarkets and analyzed for fungal contamination using conventional culture as well as molecular methods. The results of our study showed that the highest yeast and mold (YM) counts (5.34 log10 CFU g(-1)) were found in walnuts and the lowest in pecans. The most common mold in nuts was Aspergillus niger, relatively low numbers of A. flavus were found across the board, while Penicillium spp. were very common in pine nuts and walnuts. Low levels (2.00-2.84 log10 CFU g(-1)) of yeasts were recovered from only two pine nut samples. Fungal contamination in dried fruits was minimal (ranging from <2.00 to 3.86 log10 CFU g(-1)). The highest fungal levels were present in raisins. All papaya samples and the majority of cranberry, pineapple, and apricot samples were free of live fungi. The most common mold in dried fruits was A. niger followed by Penicillium spp. One apricot sample also contained low levels (2.00 log10 CFU g(-1)) of yeasts.

Fate of Shiga toxin-producing O157:H7 and non-O157:H7 Escherichia coli cells within refrigerated, frozen, or frozen then thawed ground beef patties cooked on a commercial open-flame gas or a clamshell electric grill.

PubMed

Luchansky, John B; Porto-Fett, Anna C S; Shoyer, Bradley A; Phillips, John; Chen, Vivian; Eblen, Denise R; Cook, L Victor; Mohr, Tim B; Esteban, Emilio; Bauer, Nathan

2013-09-01

Both high-fat and low-fat ground beef (percent lean:fat = ca. 70:30 and 93:7, respectively) were inoculated with a 6-strain cocktail of non-O157:H7 Shiga toxin-producing Escherichia coli (STEC) or a five-strain cocktail of E. coli O157:H7 (ca. 7.0 log CFU/g). Patties were pressed (ca. 2.54 cm thick, ca. 300 g each) and then refrigerated (4°C, 18 to 24 h), or frozen (-18°C, 3 weeks), or frozen (-18°C, 3 weeks) and then thawed (4°C for 18 h or 21°C for 10 h) before being cooked on commercial gas or electric grills to internal temperatures of 60 to 76.6°C. For E. coli O157:H7, regardless of grill type or fat level, cooking refrigerated patties to 71.1 or 76.6°C decreased E. coli O157:H7 numbers from an initial level of ca. 7.0 log CFU/g to a final level of ≤1.0 log CFU/g, whereas decreases to ca. 1.1 to 3.1 log CFU/g were observed when refrigerated patties were cooked to 60.0 or 65.5°C. For patties that were frozen or freeze-thawed and cooked to 71.1 or 76.6°C, E. coli O157:H7 numbers decreased to ca. 1.7 or ≤0.7 log CFU/g. Likewise, pathogen numbers decreased to ca. 0.7 to 3.7 log CFU/g in patties that were frozen or freeze-thawed and cooked to 60.0 or 65.5°C. For STEC, regardless of grill type or fat level, cooking refrigerated patties to 71.1 or 76.6°C decreased pathogen numbers from ca. 7.0 to ≤0.7 log CFU/g, whereas decreases to ca. 0.7 to 3.6 log CFU/g were observed when refrigerated patties were cooked to 60.0 or 65.5°C. For patties that were frozen or freeze-thawed and cooked to 71.1 or 76.6°C, STEC numbers decreased to a final level of ca. 1.5 to ≤0.7 log CFU/g. Likewise, pathogen numbers decreased from ca. 7.0 to ca. 0.8 to 4.3 log CFU/g in patties that were frozen or freeze-thawed and cooked to 60.0 or 65.5°C. Thus, cooking ground beef patties that were refrigerated, frozen, or freeze-thawed to internal temperatures of 71.1 and 76.6°C was effective for eliminating ca. 5.1 to 7.0 log CFU of E. coli O157:H7 and STEC per g.

Chitosan inactivates spoilage yeasts but enhances survival of Escherichia coli O157:H7 in apple juice.

PubMed

Kiskó, G; Sharp, R; Roller, S

2005-01-01

To develop new measures for controlling both spoilage and pathogenic micro-organisms in unpasteurized apple juice using chitosan. Micro-organisms were isolated and identified from apple juice treated or untreated with chitosan using enrichment, selective media, microscopy, substrate assimilation patterns and ribosomal DNA profiling. Chitosan (0.05-0.1%) delayed spoilage by yeasts at 25 degrees C for up to 12 days but the effect was species specific: Kloeckera apiculata and Metschnikowia pulcherrima were inactivated but Saccharomyces cerevisiae and Pichia spp. multiplied slowly. In challenge experiments at 25 degrees C, total yeast counts were 3-5 log CFU ml(-1) lower in chitosan-treated juices than in the controls for 4 days but the survival of Escherichia coli O157:H7 was extended from 1 to 2 days; at 4 degrees C, chitosan reduced the yeast counts by 2-3 log CFU ml(-1) for up to 10 days but survival of the pathogen was prolonged from 3 to 5 days. The survival of Salmonella enterica serovar Typhimurium was unaffected by chitosan at either temperature. The addition of chitosan to apple juice delayed spoilage by yeasts but enhanced the survival of E. coli O157:H7. The results suggest that the use of chitosan in the treatment of fruit juices may potentially lead to an increased risk of food poisoning from E. coli O157:H7.

Fluorocycline TP-271 Is Potent against Complicated Community-Acquired Bacterial Pneumonia Pathogens

PubMed Central

Fyfe, Corey; O’Brien, William; Hackel, Meredith; Minyard, Mary Beth; Waites, Ken B.; Dubois, Jacques; Murphy, Timothy M.; Slee, Andrew M.; Weiss, William J.; Sutcliffe, Joyce A.

2017-01-01

ABSTRACT TP-271 is a novel, fully synthetic fluorocycline antibiotic in clinical development for the treatment of respiratory infections caused by susceptible and multidrug-resistant pathogens. TP-271 was active in MIC assays against key community respiratory Gram-positive and Gram-negative pathogens, including Streptococcus pneumoniae (MIC90 = 0.03 µg/ml), methicillin-sensitive Staphylococcus aureus (MSSA; MIC90 = 0.25 µg/ml), methicillin-resistant S. aureus (MRSA; MIC90 = 0.12 µg/ml), Streptococcus pyogenes (MIC90 = 0.03 µg/ml), Haemophilus influenzae (MIC90 = 0.12 µg/ml), and Moraxella catarrhalis (MIC90 ≤0.016 µg/ml). TP-271 showed activity (MIC90 = 0.12 µg/ml) against community-acquired MRSA expressing Panton-Valentine leukocidin (PVL). MIC90 values against Mycoplasma pneumoniae, Legionella pneumophila, and Chlamydia pneumoniae were 0.004, 1, and 4 µg/ml, respectively. TP-271 was efficacious in neutropenic and immunocompetent animal pneumonia models, generally showing, compared to the burden at the start of dosing, ~2 to 5 log10 CFU reductions against MRSA, S. pneumoniae, and H. influenzae infections when given intravenously (i.v.) and ~1 to 4 log10 CFU reductions when given orally (p.o.). TP-271 was potent against key community-acquired bacterial pneumonia (CABP) pathogens and was minimally affected, or unaffected, by tetracycline-specific resistance mechanisms and fluoroquinolone or macrolide drug resistance phenotypes. IMPORTANCE Rising resistance rates for macrolides, fluoroquinolones, and β-lactams in the most common pathogens associated with community-acquired bacterial pneumonia (CABP) are of concern, especially for cases of moderate to severe infections in vulnerable populations such as the very young and the elderly. New antibiotics that are active against multidrug-resistant Streptococcus pneumoniae and Staphylococcus aureus are needed for use in the empirical treatment of the most severe forms of this disease. TP-271 is a promising new fluorocycline antibiotic demonstrating in vitro potency and nonclinical efficacy by intravenous and oral administration against the major pathogens associated with moderate to severe CABP. PMID:28251179

In vitro reduction of endotoxin concentrations with the 5S fragment of immunoglobulin G.

PubMed Central

Xuan, D; Nicolau, D P; Tessier, P R; Bow, L; Quintiliani, R; Nightingale, C H

1997-01-01

Endotoxin has long been implicated as an inducer for the development and progression of gram-negative sepsis. Accordingly, antiendotoxin therapy has been considered one of the major targets for the treatment of sepsis. To investigate the influence of a human immunoglobulin G (IgG) derivative, the 5S fragment of IgG (5S-IgG; Gamma-Venin, Centeon Pharma GmbH, Frankfurt-Niederrad, Germany), on endotoxin release during bacterial proliferation and under antibiotic bactericidal action, time-kill studies were performed by using Escherichia coli ATCC 25922 starting inocula of 10(3), 10(5), and 10(7) CFU/ml with cefotaxime (120 microg/ml) alone and in combination with 5S-IgG (2,100 microg/ml). Samples were collected for bacterial colony count and endotoxin concentration determinations; the area under the free endotoxin concentration curve (AUFEC) was calculated by using the trapezoidal rule. Colony counts showed that cefotaxime had a rapid bactericidal effect because it achieved greater than a 4-log decrease in the numbers of E. coli CFU per milliliter over the first 2 h; the addition of 5S-IgG did not appear to alter the kinetics of killing. Comparison of the AUFEC revealed that the addition of 5S-IgG resulted in a mean reduction of 50, 66, and 27% in the free endotoxin concentration at starting inocula of 10(3), 10(5), and 10(7) CFU/ml, respectively. Moreover, experiments were conducted with a starting inoculum of 10(5) CFU/ml and various amounts of 5S-IgG (2 to 20 mg/ml) to further investigate the dose-effect relation of 5S-IgG on endotoxin release. Decreased AUFECs were observed with increasing concentrations of 5S-IgG, suggesting the dose-dependent antiendotoxin activity of 5S-IgG. Further study is required to investigate the mechanism(s) responsible for this observation, the biological significance of this antiendotoxin activity, and the potential utility of 5S-IgG as an adjuvant therapy in the treatment of gram-negative sepsis. PMID:9210676

Effects of electrolyzed oxidizing water treatment on reducing Vibrio parahaemolyticus and Vibrio vulnificus in raw oysters.

PubMed

Ren, Tingting; Su, Yi-Cheng

2006-08-01

Contamination of Vibrio parahaemolyticus and Vibrio vulnificus in oysters is a food safety concern. This study investigated effects of electrolyzed oxidizing (EO) water treatment on reducing V. parahaemolyticus and V. vulnificus in laboratory-contaminated oysters. EO water exhibited strong antibacterial activity against V. parahaemolyticus and V. vulnificus in pure cultures. Populations of V. parahaemolyticus (8.74 x 10(7) CFU/ml) and V. vulnificus (8.69 x 10(7) CFU/ml) decreased quickly in EO water containing 0.5% NaCl to nondetectable levels (> 6.6 log reductions) within 15 s. Freshly harvested Pacific oysters were inoculated with a five-strain cocktail of V. parahaemolyticus or V. vulnificus at levels of 10(4) and 10(6) most probable number (MPN)/g and treated with EO water (chlorine, 30 ppm; pH 2.82; oxidation-reduction potential, 1131 mV) containing 1% NaCl at room temperature. Reductions of V. parahaemolyticus and V. vulnificus in oysters were determined at 0 (before treatment), 2, 4, 6, and 8 h of treatment. Holding oysters inoculated with V. parahaemolyticus or V. vulnificus in the EO water containing 1% NaCl for 4 to 6 h resulted in significant (P < 0.05) reductions of V. parahaemolyticus and V. vulnificus by 1.13 and 1.05 log MPN/g, respectively. Extended exposure (> 12 h) of oysters in EO water containing high levels of chlorine (> 30 ppm) was found to be detrimental to oysters. EO water could be used as a postharvest treatment to reduce Vibrio contamination in oysters. However, treatment should be limited to 4 to 6 h to avoid death of oysters. Further studies are needed to determine effects of EO water treatment on sensory characteristics of oysters.

Inactivation of Escherichia coli O157:H7 in moisture-enhanced nonintact beef by pan-broiling or roasting with various cooking appliances set at different temperatures.

PubMed

Shen, Cangliang; Geornaras, Ifigenia; Belk, Keith E; Smith, Gary C; Sofos, John N

2011-01-01

This study evaluated inactivation of Escherichia coli O157:H7 in moisture-enhanced restructured nonintact beef cooked to 65 °C using different cooking appliances set at different temperatures. Batches (2 kg) of coarse-ground beef (approximately 5% fat) were mixed with an 8-strain composite (100 mL) of rifampicin-resistant E. coli O157:H7 (6.4 ± 0.1 log CFU/g) and a solution (100 mL) of sodium chloride plus sodium tripolyphosphate to yield concentrations (wt/wt) of 0.5% and 0.25%, respectively, in the final product. Beef portions of 2.54 cm thickness (15 cm dia) were prepared and were vacuum-packaged and frozen (-20 °C, 42 h). Partially thawed (-2.5 ± 1.0 °C) portions were pan-broiled (Presto electric skillet and Sanyo grill) or roasted (Oster toaster oven and Magic Chef kitchen oven) to 65 °C. The appliances were set at, and preheated before cooking to 149 or 204 °C (electric skillet), 149 or 218 °C (grill), 149 or 232 °C (toaster oven), and 149, 204, or 260 °C (kitchen oven). Temperatures of appliances and beef samples were monitored with thermocouples, and meat samples were analyzed for surviving microbial populations. In general, the higher the appliance temperature setting, the shorter the time needed to reach 65 °C, and the higher the edge and surface temperatures of the meat samples. Temperatures of 204 to 260 °C, regardless of appliance, resulted in greater (P < 0.05) pathogen reductions (3.3 to 5.5 log CFU/g) than those obtained at 149 °C (1.5 to 2.4 log CFU/g). The highest (P < 0.05) reduction (5.5 log CFU/g) was obtained in samples cooked in the kitchen oven set at 260 °C. The results should be useful to the food service industry for selection of effective nonintact beef cooking protocols, and for use in risk assessments for nonintact meat products. Practical Application: Results of this study should be useful for developing cooking recommendations to enhance the safety of nonintact beef products, and for use in risk assessments of such products.

Microbiological characterization of a regenerative life support system

NASA Technical Reports Server (NTRS)

Koenig, D. W.; Bruce, R. J.; Mishra, S. K.; Barta, D. J.; Pierson, D. L.

1994-01-01

A Variable Pressure Plant Growth Chamber (VPGC), at the Johnson Space Center's (JSC) ground based Regenerative Life Support Systems (RLSS) test bed, was used to produce crops of soil-grown lettuce. The crops and chamber were analyzed for microbiological diversity during lettuce growth and after harvest. Bacterial counts for the rhizosphere, spent nutrient medium, heat exchanger condensate, and atmosphere were approximately 10(exp 11) Colony Forming Units (CFU)/g, 10(exp 5) CFU/ml, 10(exp 5)CFU/ml, and 600 CFU/m sq, repectively. Pseudomonas was the predominant bacterial genus. Numbers of fungi were about 10(exp 5) CFU/g in the rhizosphere, 4-200 CFU/ml in the spent nutient medium, 110 CFU/ml in the heat exchanger condensate, and 3 CFU/cu m in the atmosphere. Fusarium and Trichoderma were the predominant fungal genera.

Quest for a Realistic In Vivo Test Method for Antimicrobial Hand-Rub Agents: Introduction of a Low-Volume Hand Contamination Procedure▿

PubMed Central

Macinga, David R.; Beausoleil, Christopher M.; Campbell, Esther; Mulberry, Gayle; Brady, Ann; Edmonds, Sarah L.; Arbogast, James W.

2011-01-01

A novel method has been developed for the evaluation of alcohol-based hand rubs (ABHR) that employs a hand contamination procedure that more closely simulates the in-use conditions of ABHR. Hands of human subjects were contaminated with 0.2 ml of a concentrated suspension of Serratia marcescens (ATCC 14756) to achieve baseline contamination between 8 and 9 log10 CFU/hand while allowing product to be applied to dry hands with minimal soil load. Evaluation of 1.5 ml of an ABHR gel containing 62% ethanol produced log10 reductions of 2.66 ± 0.96, 2.40 ± 0.50, 2.41 ± 0.61, and 2.33 ± 0.49 (means ± standard deviations) after 1, 3, 7, and 10 successive contamination/product application cycles. In a study comparing this low-volume contamination (LVC) method to ASTM E1174, product dry times were more realistic and log10 reductions achieved by the ABHR were significantly greater when LVC was employed (P < 0.05). These results indicate that a novel low-volume hand contamination procedure, which more closely represents ABHR use conditions, provides more realistic estimates of in-use ABHR efficacies. Based on the LVC method, log10 reductions produced by ABHR were strongly dependent on the test product application volume (P < 0.0001) but were not influenced by the alcohol concentration when it was within the range of 62 to 85% (P = 0.378). PMID:22003004

Biofilm growth on polyvinylchloride surface incubated in suboptimal microbial warm water and effect of sanitizers on biofilm removal post biofilm formation

USDA-ARS?s Scientific Manuscript database

An in vitro experiment was conducted to understand the nature of biofilm growth on polyvinyl chloride (PVC) surface when exposed to sub optimal quality microbial water (> 4 log10 cfu/ml) obtained from poultry drinking water source mimicking water in waterlines during the first week of poultry broodi...

Study of the antibacterial activity of electro-activated solutions of salts of weak organic acids on Salmonella enterica, Staphylococcus aureus and Listeria monocytogenes.

PubMed

Liato, Viacheslav; Labrie, Steve; Aïder, Mohammed

2017-01-01

This work assessed the antibacterial activity of electro-activated solutions of salts of weak organic acids (potassium acetate, potassium citrate and calcium lactate) on Salmonella enterica, Staphylococcus aureus and Listeria monocytogenes. This activity was compared in terms of minimal inhibitory (bactericidal) concentration to the effect of commercial acetic, citric and lactic acid at equivalent titratable acidity. Staining live/dead BacLight method was used to consider physiological state of bacteria following the evaluation of pathogenic strains during exposure to the tested solutions. The results demonstrated strong inhibitory activity of all electro-activated solutions. After 10 min of exposure to electro-activated potassium acetate, a reduction of ≥6 log CFU/ml of all bacteria was observed. The electro-activated potassium citrate demonstrated the lowest minimal inhibitory concentration. Nevertheless, its inactivation power was significantly higher than that of conjugated citric acid. Although electro-activated calcium lactate was found less effective in comparison with its conjugated acid form, after 10 min of contact with the tested pathogens, it induced a population reduction of 2.23, 2.97 and 5.57 log CFU/ml of S. aureus, L. monocytogenes and S. enterica, respectively.

Assuring growth inhibition of listerial contamination during processing and storage of traditional Greek Graviera cheese: compliance with the new European Union regulatory criteria for Listeria monocytogenes.

PubMed

Samelis, John; Giannou, Eleni; Lianou, Alexandra

2009-11-01

The current microbiological regulatory criteria in the European Union specify a maximum Listeria monocytogenes population of 100 CFU/g allowable in ready-to-eat foods provided the product will not exceed this limit throughout its shelf life. The aim of this study was to validate the manufacturing method for traditional Greek Graviera cheese produced from thermized milk. Initial challenge experiments evaluated the fate of inoculated L. monocytogenes (ca. 4 log CFU/ml, three-strain cocktail) in thermized Graviera cheese milk (TGCM; 63 degrees C for 30 s) in the presence and absence of a product-specific starter culture (SC) in vitro. Milk samples were incubated for 6 h at 37 degrees C and then for 66 h at 18 degrees C. Experiments were conducted to evaluate the fate of a cocktail of three nonpathogenic L. monocytogenes and L. innocua indicator strains inoculated (ca. 3 log CFU/g) in Graviera cheeses commercially manufactured from TGCM+SC. Cheeses were brined, ripened at 18 degrees C and 90% relative humidity for 20 days, and stored at 4 degrees C for up to day 60 under vacuum. In TGCM, L. monocytogenes increased by ca. 2 log units, whereas in TGCM+SC L. monocytogenes growth was retarded (P < 0.05) after a ca. 1-log increase within 6 h at 37 degrees C. Populations of Listeria indicator strains did not grow in TGCM+SC cheeses at any stage; they declined 10-fold in fresh cheeses within 5 days and then survived with little death thereafter. Thus, growth inhibition but not inactivation of potent natural Listeria contaminants at levels below 100 CFU/g occurs in the core of traditional Greek Graviera cheese during fermentation, ripening, and storage.

Development of PMA real-time PCR method to quantify viable cells of Pantoea agglomerans CPA-2, an antagonist to control the major postharvest diseases on oranges.

PubMed

Soto-Muñoz, Lourdes; Teixidó, Neus; Usall, Josep; Viñas, Inmaculada; Crespo-Sempere, Ana; Torres, Rosario

2014-06-16

Dilution plating is the quantification method commonly used to estimate the population level of postharvest biocontrol agents, but this method does not permit a distinction among introduced and indigenous strains. Recently, molecular techniques based on DNA amplification such as quantitative real-time PCR (qPCR) have been successfully applied for their high strain-specific detection level. However, the ability of qPCR to distinguish viable and nonviable cells is limited. A promising strategy to avoid this issue relies on the use of nucleic acid intercalating dyes, such as propidium monoazide (PMA), as a sample pretreatment prior to the qPCR. The objective of this study was to optimize a protocol based on PMA pre-treatment samples combined with qPCR to distinguish and quantify viable cells of the biocontrol agent P. agglomerans CPA-2 applied as a postharvest treatment on orange. The efficiency of PMA-qPCR method under the established conditions (30μM PMA for 20min of incubation followed by 30min of LED light exposure) was evaluated on an orange matrix. Results showed no difference in CFU or cells counts of viable cells between PMA-qPCR and dilution plating. Samples of orange matrix inoculated with a mixture of viable/dead cells showed 5.59log10 CFU/ml by dilution plating, 8.25log10 cells/ml by qPCR, and 5.93log10 cells/ml by PMA-qPCR. Furthermore, samples inoculated with heat-killed cells were not detected by dilution plating and PMA-qPCR, while by qPCR was of 8.16log10 cells/ml. The difference in quantification cycles (Cq) among qPCR and PMA-qPCR was approximately 16cycles, which means a reduction of 65,536 fold of the dead cells detected. In conclusion, PMA-qPCR method is a suitable tool for quantify viable CPA-2 cells, which could be useful to estimate the ability of this antagonist to colonize the orange surface. Copyright © 2014 Elsevier B.V. All rights reserved.

Survival of Salmonella enterica serovar infantis on and within stored table eggs.

PubMed

Lublin, Avishai; Maler, Ilana; Mechani, Sara; Pinto, Riky; Sela-Saldinger, Shlomo

2015-02-01

Contaminated table eggs are considered a primary source of foodborne salmonellosis globally. Recently, a single clone of Salmonella enterica serovar Infantis emerged in Israel and became the predominant serovar isolated in poultry. This clone is currently the most prevalent strain in poultry and is the leading cause of salmonellosis in humans. Because little is known regarding the potential transmission of this strain from contaminated eggs to humans, the objective of this study was to evaluate the ability of Salmonella Infantis to survive on the eggshell or within the egg during cold storage or at room temperature. Salmonella cells (5.7 log CFU per egg) were inoculated on the surface of 120 intact eggs or injected into the egg yolk (3.7 log CFU per egg) of another 120 eggs. Half of the eggs were stored at 5.5 ± 0.3°C and half at room temperature (25.5 ± 0.1°C) for up to 10 weeks. At both temperatures, the number of Salmonella cells on the shell declined by 2 log up to 4 weeks and remained constant thereafter. Yolk-inoculated Salmonella counts at cold storage declined by 1 log up to 4 weeks and remained constant, while room-temperature storage supported the growth of the pathogen to a level of 8 log CFU/ml of total egg content, as early as 4 weeks postinoculation. Examination of egg content following surface inoculation revealed the presence of Salmonella in a portion of the eggs at both temperatures up to 10 weeks, suggesting that this strain can also penetrate through the shell and survive within the egg. These findings imply that Salmonella enterica serovar Infantis is capable of survival both on the exterior and interior of table eggs and even multiply inside the egg at room temperature. Our findings support the need for prompt refrigeration to prevent Salmonella multiplication during storage of eggs at room temperature.

Survival of Bacillus anthracis spores in fruit juices and wine.

PubMed

Leishman, Oriana N; Johnson, Miranda J; Labuza, Theodore P; Diez-Gonzalez, Francisco

2010-09-01

Foods have been identified as a potential target for bioterrorism due to their essential nature and global distribution. Foods produced in bulk have the potential to have large batches of product intentionally contaminated, which could affect hundreds or thousands of individuals. Bacillus anthracis spores are one potential bioterrorism agent that may survive pasteurization and remain viable throughout the shelf life of fruit juices and cause disease if consumed. This project examined B. anthracis spore survival in orange, apple, and grape juices, as well as wine. Samples of beverages were inoculated with spores of two nonpathogenic B. anthracis strains at approximately 10(6) CFU/ml, and the spore count was determined periodically during storage for 30 days at 4°C. After this time, the counts of survival spores never declined more than 1 log CFU/ml in any of the beverage types. These results indicate that spores can survive, with little to no loss in viability, for at least a month in fruit juices and wine.

Synthesis of omeprazole analogues and evaluation of these as potential inhibitors of the multidrug efflux pump NorA of Staphylococcus aureus.

PubMed

Vidaillac, Céline; Guillon, Jean; Arpin, Corinne; Forfar-Bares, Isabelle; Ba, Boubakar B; Grellet, Jean; Moreau, Stéphane; Caignard, Daniel-Henri; Jarry, Christian; Quentin, Claudine

2007-03-01

A series of 11 pyrrolo[1,2-a]quinoxaline derivatives, 1a to 1k, sharing structural analogies with omeprazole, a eukaryotic efflux pump inhibitor (EPI) used as an antiulcer agent, was synthesized. Their inhibitory effect was evaluated using Staphylococcus aureus strain SA-1199B overexpressing NorA. By determinations of the MIC of norfloxacin in the presence of these EPIs devoid of intrinsic antibacterial activity and used at 128 microg/ml, and by the checkerboard method, compound 1e (MIC decrease, 16-fold; fractional inhibitory concentration index [SigmaFIC], 0.18) appeared to be more active than compounds 1b to 1d, reserpine, and omeprazole (MIC decrease, eightfold; SigmaFIC, 0.31), followed by compounds 1a and 1f (MIC decrease, fourfold; SigmaFIC, 0.37) and 1g to 1k (MIC decrease, twofold; SigmaFIC, 0.50 to 0.56). By time-kill curves combining norfloxacin (1/4 MIC) and the most efficient EPIs (128 microg/ml), compound 1e persistently restored the bactericidal activity of norfloxacin (inoculum reduction, 3 log(10) CFU/ml at 8 and 24 h), compound 1f led to a delayed but progressive decrease in the number of viable cells, and compounds 1b to 1d and omeprazole acted synergistically (inoculum reduction, 3 log(10) CFU/ml at 8 h but further regrowth), while compound 1a and reserpine slightly enhanced norfloxacin activity. The bacterial uptake of norfloxacin monitored by high-performance liquid chromatography confirmed that compounds 1a to 1f increased antibiotic accumulation, as did reserpine and omeprazole. Since these EPIs did not disturb the Deltapsi and DeltapH, they might directly interact with the pump. A structure-activity relationships study identified the benzimidazole nucleus of omeprazole as the main structural element involved in efflux pump inhibition and highlighted the critical role of the chlorine substituents in the stability and efficiency of compounds 1e to 1f. However, further pharmacomodulation is required to obtain therapeutically applicable derivatives.

Predicting acute uncomplicated urinary tract infection in women: a systematic review of the diagnostic accuracy of symptoms and signs

PubMed Central

2010-01-01

Background Acute urinary tract infections (UTI) are one of the most common bacterial infections among women presenting to primary care. However, there is a lack of consensus regarding the optimal reference standard threshold for diagnosing UTI. The objective of this systematic review is to determine the diagnostic accuracy of symptoms and signs in women presenting with suspected UTI, across three different reference standards (102 or 103 or 105 CFU/ml). We also examine the diagnostic value of individual symptoms and signs combined with dipstick test results in terms of clinical decision making. Methods Searches were performed through PubMed (1966 to April 2010), EMBASE (1973 to April 2010), Cochrane library (1973 to April 2010), Google scholar and reference checking. Studies that assessed the diagnostic accuracy of symptoms and signs of an uncomplicated UTI using a urine culture from a clean-catch or catherised urine specimen as the reference standard, with a reference standard of at least ≥ 102 CFU/ml were included. Synthesised data from a high quality systematic review were used regarding dipstick results. Studies were combined using a bivariate random effects model. Results Sixteen studies incorporating 3,711 patients are included. The weighted prior probability of UTI varies across diagnostic threshold, 65.1% at ≥ 102 CFU/ml; 55.4% at ≥ 103 CFU/ml and 44.8% at ≥ 102 CFU/ml ≥ 105 CFU/ml. Six symptoms are identified as useful diagnostic symptoms when a threshold of ≥ 102 CFU/ml is the reference standard. Presence of dysuria (+LR 1.30 95% CI 1.20-1.41), frequency (+LR 1.10 95% CI 1.04-1.16), hematuria (+LR 1.72 95%CI 1.30-2.27), nocturia (+LR 1.30 95% CI 1.08-1.56) and urgency (+LR 1.22 95% CI 1.11-1.34) all increase the probability of UTI. The presence of vaginal discharge (+LR 0.65 95% CI 0.51-0.83) decreases the probability of UTI. Presence of hematuria has the highest diagnostic utility, raising the post-test probability of UTI to 75.8% at ≥ 102 CFU/ml and 67.4% at ≥ 103 CFU/ml. Probability of UTI increases to 93.3% and 90.1% at ≥ 102 CFU/ml and ≥ 103 CFU/ml respectively when presence of hematuria is combined with a positive dipstick result for nitrites. Subgroup analysis shows improved diagnostic accuracy using lower reference standards ≥ 102 CFU/ml and ≥ 103 CFU/ml. Conclusions Individual symptoms and signs have a modest ability to raise the pretest-risk of UTI. Diagnostic accuracy improves considerably when combined with dipstick tests particularly tests for nitrites. PMID:20969801

Efficacy of commercial produce sanitizers against nontoxigenic Escherichia coli O157:H7 during processing of iceberg lettuce in a pilot-scale leafy green processing line.

PubMed

Davidson, Gordon R; Buchholz, Annemarie L; Ryser, Elliot T

2013-11-01

Chemical sanitizers are routinely used during commercial flume washing of fresh-cut leafy greens to minimize cross-contamination from the water. This study assessed the efficacy of five commercial sanitizer treatments against Escherichia coli O157:H7 on iceberg lettuce, in wash water, and on equipment during simulated commercial production in a pilot-scale processing line. Iceberg lettuce (5.4 kg) was inoculated to contain 10(6) CFU/g of a four-strain cocktail of nontoxigenic, green fluorescent protein-labeled, ampicillin-resistant E. coli O157:H7 and processed after 1 h of draining at ~22 °C. Lettuce was shredded using a commercial slicer, step-conveyed to a flume tank, washed for 90 s using six different treatments (water alone, 50 ppm of peroxyacetic acid, 50 ppm of mixed peracid, or 50 ppm of available chlorine either alone or acidified to pH 6.5 with citric acid [CA] or T-128), and then dried using a shaker table and centrifugal dryer. Various product (25-g) and water (50-ml) samples collected during processing along with equipment surface samples (100 cm(2)) from the flume tank, shaker table, and centrifugal dryer were homogenized in neutralizing buffer and plated on tryptic soy agar. During and after iceberg lettuce processing, none of the sanitizers were significantly more effective (P ≤ 0.05) than water alone at reducing E. coli O157:H7 populations on lettuce, with reductions ranging from 0.75 to 1.4 log CFU/g. Regardless of the sanitizer treatment used, the centrifugal dryer surfaces yielded E. coli O157:H7 populations of 3.49 to 4.98 log CFU/100 cm(2). Chlorine, chlorine plus CA, and chlorine plus T-128 were generally more effective (P ≤ 0.05) than the other treatments, with reductions of 3.79, 5.47, and 5.37 log CFU/ml after 90 s of processing, respectively. This indicates that chlorine-based sanitizers will likely prevent wash water containing low organic loads from becoming a vehicle for cross-contamination.

Microbiological survey of five poultry processing plants in the UK.

PubMed

Mead, G C; Hudson, W R; Hinton, M H

1993-07-01

1. Neck skin samples were taken from chickens and turkeys at all the main stages of processing to monitor changes in total viable count (TVC) and counts of coliforms and pseudomonads. 2. Processing reduced TVC by up to 100-fold. Geometric mean counts after packaging were log10 4.4 to 5.3 CFU/g whilst corresponding counts of coliforms were 2.7 to 3.8 CFU/g. 3. Increases in mean TVC or coliforms as a result of either defeathering or evisceration did not exceed 0.6 log. 4. Pseudomonads represented only a minor fraction of the initial microflora of the bird and were often reduced by scalding to a figure which could not be detected by direct plating of samples; however, subsequent contamination resulted in means between log10 2.9 and 4.0 CFU/g for packaged carcases. 5. Although Staphylococcus aureus was readily isolated from defeathering equipment, mean counts from defeathered carcases were always below log10 3.0 CFU/g.

Fate of Escherichia coli O157 Cells Inoculated into Lightly Pickled Chinese Cabbage during Processing, Storage and Incubation in Artificial Gastric Juice.

PubMed

Inatsu, Yasuhiro; Ohata, Yukiko; Ananchaipattana, Chiraporn; Latiful Bari, Md; Hosotani, Yukie; Kawasaki, Susumu

2016-01-01

Fate of Escherichia coli O157 cells was evaluated when inoculated into each step after production of lightly pickled Chinese cabbage. The efficacy of surface sterilization by 100 mg/L of chlorine water for 10 min on raw leaves (6.0 log CFU/g) was 2.2 log CFU/g reduction. No meaningful change of the population of E. coli O157 (3.5 log CFU/g to 1.5 log MPN/g) contaminated into 19 kinds of products was observed. These results indicated the difficulty of estimating the viable count of the cells between contaminated on farms and further processing and storage steps. The population of E. coli O157 (3 log CFU/g to 1 log MPN/g) inoculated into the Chinese cabbage products was reduced less than 0.6 log CFU/g after 2 h-incubation at 37℃ in artificial gastric juice. Prevention from initial contamination of E. coli O157 on the ingredients of Chinese cabbage products is important to reduce the risk of food poisoning because the reduction of the bacterial counts after processing and consumption are limited.

Effect of inoculation with Penicillium expansum on the microbial community and maturity of compost.

PubMed

Wang, Hong-yuan; Fan, Bing-quan; Hu, Qing-xiu; Yin, Zhong-wei

2011-12-01

Compost prepared from wheat straw and cattle/chicken mature was inoculated with the lignocellulolytic fungus, Penicillium expansum. Compared to uninoculated compost, the inoculated compost exhibited a 150% higher germination index, more than 1.2 g kg(-1)-dw of changes in NH(4)(+)-N concentrations, a ca. 12.0% higher humus content and a lignocellulose degradation that proceeded 57.5% faster. Culture-based determinations of microbial populations demonstrated that aerobic heterotrophic bacteria and fungi were about 1-2 orders of magnitude higher in inoculated than in uninoculated compost. The number of ammonifying, ammonium-oxidizing, nitrite-oxidizing, denitrifying bacteria and cellulose-decomposing bacteria was 6.1-9.0 log(10) CFU g(-1)-dw, 1.2-4.3 log(10) MPN g(-1)-dw, 3.5-6.8 log(10) MPN g(-1)-dw, 3.58-4.34 log(10) MPN g(-1)-dw, 1.4-3.8 log(10)MPN g(-1)-dw, and 4.2-8.8 log(10) CFU g(-1)-dw higher in the compost inoculated with P. expansum. Copyright © 2011 Elsevier Ltd. All rights reserved.

Application of quantitative real-time PCR compared to filtration methods for the enumeration of Escherichia coli in surface waters within Vietnam.

PubMed

Vital, Pierangeli G; Van Ha, Nguyen Thi; Tuyet, Le Thi Hong; Widmer, Kenneth W

2017-02-01

Surface water samples in Vietnam were collected from the Saigon River, rural and suburban canals, and urban runoff canals in Ho Chi Minh City, Vietnam, and were processed to enumerate Escherichia coli. Quantification was done through membrane filtration and quantitative real-time polymerase chain reaction (PCR). Mean log colony-forming unit (CFU)/100 ml E. coli counts in the dry season for river/suburban canals and urban canals were log 2.8 and 3.7, respectively, using a membrane filtration method, while using Taqman quantitative real-time PCR they were log 2.4 and 2.8 for river/suburban canals and urban canals, respectively. For the wet season, data determined by the membrane filtration method in river/suburban canals and urban canals samples had mean counts of log 3.7 and 4.1, respectively. While mean log CFU/100 ml counts in the wet season using quantitative PCR were log 3 and 2, respectively. Additionally, the urban canal samples were significantly lower than those determined by conventional culture methods for the wet season. These results show that while quantitative real-time PCR can be used to determine levels of fecal indicator bacteria in surface waters, there are some limitations to its application and it may be impacted by sources of runoff based on surveyed samples.

A fully automated microfluidic-based electrochemical sensor for real-time bacteria detection.

PubMed

Altintas, Zeynep; Akgun, Mete; Kokturk, Guzin; Uludag, Yildiz

2018-02-15

A fully automated microfluidic-based electrochemical biosensor was designed and manufactured for pathogen detection. The quantification of Escherichia coli was investigated with standard and nanomaterial amplified immunoassays in the concentration ranges of 0.99 × 10 4 3.98 × 10 9 cfu mL -1 and 103.97 × 10 7 cfu mL -1 which resulted in detection limits of 1.99 × 10 4 cfu mL -1 and 50 cfu mL -1 , respectively. The developed methodology was then applied for E. coli quantification in water samples using nanomaterial modified assay. Same detection limit for E. coli was achieved for real sample analysis with a little decrease on the sensor signal. Cross-reactivity studies were conducted by testing Shigella, Salmonella spp., Salmonella typhimurium and Staphylococcus aureus on E. coli specific antibody surface that confirmed the high specificity of the developed immunoassays. The sensor surface could be regenerated multiple times which significantly reduces the cost of the system. Our custom-designed biosensor is capable of detecting bacteria with high sensitivity and specificity, and can serve as a promising tool for pathogen detection. Copyright © 2017 Elsevier B.V. All rights reserved.

Antimicrobial Efficacy of a Sulfuric Acid and Sodium Sulfate Blend, Peroxyacetic Acid, and Cetylpyridinium Chloride against Salmonella on Inoculated Chicken Wings.

PubMed

Scott, Brittney R; Yang, Xiang; Geornaras, Ifigenia; Delmore, Robert J; Woerner, Dale R; Reagan, James O; Morgan, J Brad; Belk, Keith E

2015-11-01

Studies were conducted to evaluate the efficacy of a commercial blend of sulfuric acid and sodium sulfate (SSS) in reducing Salmonella on inoculated whole chilled chicken wings and to compare its efficacy to peroxyacetic acid (PAA) and cetylpyridinium chloride (CPC). Wings were spot inoculated (5 to 6 log CFU/ml of sample rinsate) with a five-strain mixture of novobiocin- and nalidixic acid-resistant Salmonella and then left untreated (control) or treated by immersing individual wings in 350 ml of antimicrobial solution. An initial study evaluated two treatment immersion times, 10 and 20 s, of SSS (pH 1.1) and compared cell recoveries following rinsing of treated samples with buffered peptone water or Dey/Engley neutralizing broth. In a second study, inoculated wings were treated with SSS (pH 1.1; 20 s), PAA (700 ppm, 20 s), or CPC (4,000 ppm, 10 s) and analyzed for survivors immediately after treatment (0 h) and after 24 h of aerobic storage at 4°C. Color and pH analyses were also conducted in the latter study. Recovery of Salmonella survivors following treatment with SSS (10 or 20 s) was not (P ≥ 0.05) affected by the type of cell recovery rinse solution (buffered peptone water or Dey/Engley neutralizing broth), but there was an effect (P < 0.05) of SSS treatment time. Immersion of samples for 10 or 20 s in SSS resulted in pathogen reductions of 0.8 to 0.9 and 1.1 to 1.2 log CFU/ml, respectively. Results of the second study showed that there was an interaction (P < 0.05) between antimicrobial type and storage time. Efficacy against Salmonella at 0 h increased in the order CPC , SSS , PAA; however, after 24 h of aerobic storage, pathogen counts of SSS- and PAA-treated wings did not differ (P ≥ 0.05). Overall, the results indicated that SSS applied at pH 1.1 for 20 s was an effective antimicrobial intervention to reduce Salmonella contamination on chicken wings.

[Polymerase chain reaction (PCR) for the identification of toxigenic Vibrio cholerae O1 in oysters].

PubMed

Rodríguez-Angeles, M G; Giono-Cerezo, S; Moreno-Escobar, A; Valdespino-Gómez, J L

1994-01-01

PCR was made with ctx2 (CGG GCA GAT TCT AGA CCT CCT G) y ctx3 (CGA TGA TCT TGG AGC ATT CCC AC) primers for subunit A of cholera toxin, 30 cycles of temperature on samples of 50 g of oysters added in 450 ml of peptone alcaline water that were inoculated with 15 x 10(6), 0.75 x 10(6) and 0.15 x 10(6) CFU/ml of toxigenic 6707 V. cholerae O1 reference strain. The samples were tested by three microbiological methods: INDRE's method uses 1 x 10(-1) dilution of sample, two fold pass to peptone alcaline water pH 9 incubated 18 h and 6 h at 37 degrees C, the Food and Drugs Administration (FDA) method uses 10(-1) to 10(-6) dilutions of sample, 6 h incubation and reincubation for 18 h at 37 and 42 degrees C and the Mexican laboratories (LMD) with 10(-4) to 10(-3) dilutions, the samples were incubated for 6 h and then reincubated for 18 h at two temperatures 37 and 42 degrees C. The PCR by INDRE's method was positive with 3 x 10(2) CFU/ml/g oyster. In the FDA's method the PCR detected DNA in 10(-4) dilution with 3 x 10(1) CFU/ml/g oyster and in LMD's method the PCR was positive in 10(-3) with 3 CFU/ml/g oyster. The results of the PCR were obtained between 5-6 h, and later V. cholerae O1 was isolated by three microbiological methods. The PCR reproducibility was better on DNA sample diluted 1:4 and 10 microliters of sample increased from 1:1000 to 1:10000 the sensitivity of PCR.

Impact of different tongue cleaning methods on the bacterial load of the tongue dorsum.

PubMed

Bordas, Alice; McNab, Rod; Staples, Angela M; Bowman, Jim; Kanapka, Joe; Bosma, Marylynn P

2008-04-01

To assess the extent and duration of the effect of tongue cleaning procedures on bacterial load on the dorsal surface of the tongue. 19 subjects participated in this blinded crossover study. Subjects abstained from oral hygiene, eating and drinking from 22:00 h the previous evening. Tongue samples were collected at baseline and within 15 minutes of one of three procedures: teeth brushing alone; teeth brushing plus tongue scraping; teeth brushing plus tongue cleaning using a high speed vacuum ejector and irrigation with 20 ml antibacterial mouthwash. Subjects then brushed twice daily for 3 days apart from the second group who additionally scraped their tongue twice daily. On day 4, baseline and post-treatment samples were collected as per day 1. Bacteria (total anaerobes, Gram-negative anaerobes, VSC-producing bacteria and Streptococcus saliuarius) were enumerated using appropriate selective media. The tongue dorsum was colonized by all 4 bacterial categories (log(10) 6-8 cfu/sample). For subjects who brushed their teeth only, there was a significant reduction from baseline for S. saliuarius only. In contrast, tooth brushing plus tongue scraping resulted in statistically significant reductions from baseline for all bacterial categories (range log(10) 0.11-0.40 cfu/sample). Highly statistically significant reductions (log(10) 1.11-1.96 cfu/sample) were observed for subjects who underwent thorough tongue cleaning with the saliva ejector/mouthwash. To determine longevity of treatment effects, baseline bacterial loads for days 1 and 4 were compared. Only daily tongue scraping resulted in statistical significant reduction in baseline microbial loads on day 4. While mechanical tongue cleaning with or without chemical intervention can reduce bacterial load on the tongue, this effect is transient, and regular tongue cleaning is required to provide a long lasting (overnight) reduction in bacterial numbers. Nevertheless, tongue cleaning is an oral hygiene procedure that is little practiced due to discomfort and/or lack of awareness on the part of dental professionals and their patients.

Pathogenic Gut Flora in Patients With Chronic Heart Failure.

PubMed

Pasini, Evasio; Aquilani, Roberto; Testa, Cristian; Baiardi, Paola; Angioletti, Stefania; Boschi, Federica; Verri, Manuela; Dioguardi, Francesco

2016-03-01

The goal of this study was to measure the presence of pathogenic gut flora and intestinal permeability (IP) and their correlations with disease severity, venous blood congestion, and inflammation in patients with chronic heart failure (CHF). Evidence suggests that translocation of gut flora and/or their toxins from the intestine to the bloodstream is a possible trigger of systemic CHF inflammation. However, the relation between pathogenic gut flora and CHF severity, as well as IP, venous blood congestion as right atrial pressure (RAP), and/or systemic inflammation (C-reactive protein [CRP]), is still unknown. This study analyzed 60 well-nourished patients in stable condition with mild CHF (New York Heart Association [NYHA] functional class I to II; n = 30) and moderate to severe CHF (NYHA functional class III to IV; n = 30) and matched healthy control subjects (n = 20). In all subjects, the presence and development in the feces of bacteria and fungi (Candida species) were measured; IP according to cellobiose sugar test results was documented. The study data were then correlated with RAP (echocardiography) and systemic inflammation. Compared with normal control subjects, the entire CHF population had massive quantities of pathogenic bacteria and Candida such as Campylobacter (85.3 ± 3.7 CFU/ml vs. 1.0 ± 0.3 CFU/ml; p < 0.001), Shigella (38.9 ± 12.3 CFU/ml vs. 1.6 ± 0.2 CFU/ml; p < 0.001), Salmonella (31.3 ± 9.1 CFU/ml vs 0 CFU/ml; p < 0.001), Yersinia enterocolitica (22.9 ± 6.3 CFU/ml vs. 0 CFU/ml; p < 0.0001), and Candida species (21.3 ± 1.6 CFU/ml vs. 0.8 ± 0.4 CFU/ml; p < 0.001); altered IP (10.2 ± 1.2 mg vs. 1.5 ± 0.8 mg; p < 0.001); and increased RAP (12.6 ± 0.6 mm Hg) and inflammation (12.5 ± 0.6 mg/dl). These variables were more pronounced in patients with moderate to severe NYHA functional classes than in patients with the mild NYHA functional class. Notably, IP, RAP, and CRP were mutually interrelated (IP vs. RAP, r = 0.55; p < 0.0001; IP vs. CRP, r = 0.78; p < 0.0001; and RAP vs. CRP, r = 0.78; p < 0.0001). This study showed that patients with CHF may have intestinal overgrowth of pathogenic bacteria and Candida species and increased IP associated with clinical disease severity, venous blood congestion, and inflammation. Copyright © 2016 American College of Cardiology Foundation. Published by Elsevier Inc. All rights reserved.

Effect of inoculum size, bacterial species, type of surfaces and contact time to the transfer of foodborne pathogens from inoculated to non-inoculated beef fillets via food processing surfaces.

PubMed

Gkana, E; Chorianopoulos, N; Grounta, A; Koutsoumanis, K; Nychas, G-J E

2017-04-01

The objective of the present study was to determine the factors affecting the transfer of foodborne pathogens from inoculated beef fillets to non-inoculated ones, through food processing surfaces. Three different levels of inoculation of beef fillets surface were prepared: a high one of approximately 10 7  CFU/cm 2 , a medium one of 10 5  CFU/cm 2 and a low one of 10 3  CFU/cm 2 , using mixed-strains of Listeria monocytogenes, or Salmonella enterica Typhimurium, or Escherichia coli O157:H7. The inoculated fillets were then placed on 3 different types of surfaces (stainless steel-SS, polyethylene-PE and wood-WD), for 1 or 15 min. Subsequently, these fillets were removed from the cutting boards and six sequential non-inoculated fillets were placed on the same surfaces for the same period of time. All non-inoculated fillets were contaminated with a progressive reduction trend of each pathogen's population level from the inoculated fillets to the sixth non-inoculated ones that got in contact with the surfaces, and regardless the initial inoculum, a reduction of approximately 2 log CFU/g between inoculated and 1st non-inoculated fillet was observed. S. Typhimurium was transferred at lower mean population (2.39 log CFU/g) to contaminated fillets than E. coli O157:H7 (2.93 log CFU/g), followed by L. monocytogenes (3.12 log CFU/g; P < 0.05). Wooden surfaces (2.77 log CFU/g) enhanced the transfer of bacteria to subsequent fillets compared to other materials (2.66 log CFU/g for SS and PE; P < 0.05). Cross-contamination between meat and surfaces is a multifactorial process strongly depended on the species, initial contamination level, kind of surface, contact time and the number of subsequent fillet, according to analysis of variance. Thus, quantifying the cross-contamination risk associated with various steps of meat processing and food establishments or households can provide a scientific basis for risk management of such products. Copyright © 2016 Elsevier Ltd. All rights reserved.

Room temperature sterilization of surfaces and fabrics with a one atmosphere uniform glow discharge plasma.

PubMed

Kelly-Wintenberg, K; Montie, T C; Brickman, C; Roth, J R; Carr, A K; Sorge, K; Wadsworth, L C; Tsai, P P

1998-01-01

We report the results of an interdisciplinary collaboration formed to assess the sterilizing capabilities of the One Atmosphere Uniform Glow Discharge Plasma (OAUGDP). This newly-invented source of glow discharge plasma (the fourth state of matter) is capable of operating at atmospheric pressure in air and other gases, and of providing antimicrobial active species to surfaces and workpieces at room temperature as judged by viable plate counts. OAUGDP exposures have reduced log numbers of bacteria, Staphylococcus aureus and Escherichia coli, and endospores from Bacillus stearothermophilus and Bacillus subtilis on seeded solid surfaces, fabrics, filter paper, and powdered culture media at room temperature. Initial experimental data showed a two-log10 CFU reduction of bacteria when 2 x 10(2) cells were seeded on filter paper. Results showed > or = 3 log10 CFU reduction when polypropylene samples seeded with E. coli (5 x 10(4)) were exposed, while a 30 s exposure time was required for similar killing with S. aureus-seeded polypropylene samples. The exposure times required to effect > or = 6 log10 CFU reduction of E. coli and S. aureus on polypropylene samples were no longer than 30 s. Experiments with seeded samples in sealed commercial sterilization bags showed little or no differences in exposure times compared to unwrapped samples. Plasma exposure times of less than 5 min generated > or = 5 log10 CFU reduction of commercially prepared Bacillus subtilis spores (1 x 10(5)); 7 min OAUGDP exposures were required to generate a > or = 3 log10 CFU reduction for Bacillus stearothermophilus spores. For all microorganisms tested, a biphasic curve was generated when the number of survivors vs time was plotted in dose-response cures. Several proposed mechanisms of killing at room temperature by the OAUGDP are discussed.

Mitigation of Alicyclobacillus spp. spores on food contact surfaces with aqueous chlorine dioxide and hypochlorite.

PubMed

Friedrich, Loretta M; Goodrich-Schneider, Renee; Parish, Mickey E; Danyluk, Michelle D

2009-12-01

The prevalence of Alicyclobacillus spp. and other spore-forming spoilage organisms in food handling and processing environments presents a sanitation challenge to manufacturers of products such as juices and beverages. The objectives of this study were to determine the efficacy of chlorine dioxide and sodium hypochlorite in killing Alicyclobacillus spores in situ and to evaluate the efficacy of various chlorine dioxide and hypochlorite sanitizing regimes on Alicyclobacillus spp. spores on stainless steel, wood, and rubber conveyor material. Five or two log CFU/ml spore concentrations were left in aqueous solution or inoculated onto stainless steel, rubber, or wood coupons and challenged with sanitizer for varied time intervals. After treatment, the coupons were placed in sterile sample bags, massaged with neutralizing buffer, and enumerated on Ali agar. Surfaces were also examined before and after treatment by scanning electron microscopy to confirm destruction or removal of the spores. For both five and two log CFU/ml spore concentrations, treatments of 50 and 100 ppm of chlorine dioxide and 1000 and 2000 ppm of hypochlorite, respectively, were the most effective. Of the range of chlorine dioxide concentrations and contact time regimes evaluated for all surfaces, the most effective concentration/time regime applied was 100 ppm for 10 min. Reductions ranged from 0 to 4.5 log CFU/coupon. Chlorine dioxide was least effective when applied to wood. Hypochlorite was not efficient at eliminating Alicyclobacillus spores from any of the food contact surfaces at any time and concentration combinations tested. Chlorine dioxide is an alternative treatment to kill spores of Alicyclobacillus spp. in the processing environment.

The effect of different cooking procedures on microbiological and chemical quality characteristics of Tekirdağ meatballs.

PubMed

Yilmaz, I; Yetim, H; Ockerman, H W

2002-08-01

In this research, the effects of different cooking processes (grilling, oven, and microwave cooking) on microbial flora and chemical composition of the raw and cooked meatballs as consumed in Tekirdağ were investigated. Microbial flora of the raw meatballs was as follows: total bacteria, 6.02 x 10(6) cfu/g; psychrophilic bacteria, 1.3 x 10(5) cfu/g; yeast and mould, 2.4 x 10(5) cfu/g; coliforms, 1.1 x 10(5) cfu/g; Escherichia coli, 1.0 x 10(2) cfu/g; total staphylococcae, 3.3 x 10(2) cfu/g; Staphylococcus aureus, 85 cfu/g. While Salmonella was found in only one sample, none of the samples contained Clostridium perfringens. The cooking processes clearly decreased the microbial flora (2-3 log cycles in grilling (71 degrees C) and oven-cooked (79 degrees C), 3-4 log cycles in microwave (97 degrees C) heating) of the meatballs. However, because of the crust formation and high moisture losses from the meatball surface in microwave heating, some sensorial defects were observed in the final product. Also, fat and moisture losses were higher in microwave cooking compared to the other cooking processes. In conclusion, it is advised to use slightly higher temperatures than used in the grilling or conventinal cooking procedures to increase microbial quality of the meatballs studied in this research.

The Inoculum Effect in the Era of Multidrug Resistance: Minor Differences in Inoculum Have Dramatic Effect on Minimal Inhibitory Concentration Determination.

PubMed

Smith, Kenneth P; Kirby, James E

2018-05-21

The observed MIC may depend on the number of bacteria initially inoculated into the assay. This phenomenon is termed the inoculum effect (IE) and is often most pronounced for β-lactams in strains expressing β-lactamase enzymes. The Clinical and Laboratory Standards Institute (CLSI) recommended inoculum is 5 x 10 5 CFU mL -1 with an acceptable range of 2-8 x 10 5 CFU mL -1 IE testing is typically performed using an inoculum 100-fold greater than the CLSI recommended inoculum. Therefore, it remains unknown whether the IE influences MICs during testing performed according to CLSI guidelines. Here, we utilized inkjet printing technology to test the IE on cefepime, meropenem, and ceftazidime-avibactam. First, we determined that inkjet dispense volume correlated well with the number of bacteria delivered to microwells in two-fold (R 2 = 0.99) or 1.1-fold (R 2 = 0.98) serial dilutions. We then quantified the IE by dispensing orthogonal titrations of bacterial cells and antibiotics. For cefepime resistant and susceptible dose-dependent strains, a 2-fold increase in inoculum resulted in a 1.6 Log 2 -fold increase in MIC. For carbapenemase-producing strains, each 2-fold reduction in inoculum resulted in a 1.26 Log 2 -fold reduction in meropenem MIC. At the lower end of the CLSI allowable inoculum range, minor error rates of 34.8% were observed for meropenem when testing a resistant strain set. Ceftazidime-avibactam was not subject to an appreciable IE. Our results suggest that IE is sufficiently pronounced for meropenem and cefepime in multidrug-resistant Gram-negative pathogens to affect categorical interpretations during standard laboratory testing. Copyright © 2018 American Society for Microbiology.

Correlation of E. coli K-1 bacteremia and capsular polysaccharide antigenemia in acute and chronic infection.

PubMed

Stevens, P; Young, L S; Alam, S

1983-09-01

The K-1 polysaccharide is an important virulence factor in human E. coli infections. Using E. coli 016K1, we investigated the kinetic association of bacteremia and K-1 antigenemia in acute lapine and canine infections and in a chronic infection model of neutropenic rats. Additionally, we assessed the presence of K-1 antigenemia in E. coli K-1 bacteremic patients. K-1 was measured by a solid phase radioimmunoassay (RIA) using cross-reactive equine anti-Group B meningococcal IgM. In acute infections, none of the dogs or rabbits developed antigenemia even with a bacteremia of 2 X 10(4) CFU/ml or 5 X 10(5) CFU/ml, respectively. Antigenemia appeared in the rabbit only with an infecting dose of greater than or equal to 5 X 10(8) CFU. In the rat model we observed an initial bacteremia of 10(3) CFU/ml, which increased to 10(6) CFU/ml at 24 hrs. However, antigenemia was most often delayed, appearing in only greater than or equal to 30 hrs postinfection. Percent mortality was directly associated with the degree of bacteremia and antigenemia. In acute human E. coli K-1 bacteremia, 11 of 22 (50%) of patients were positive for K-1 antigenemia. The data demonstrated that K-1 polysaccharide was not usually detectable in the early stages of bacteremia, but occurred only after prolonged infection or very high infecting doses. The RIA to measure K-1 antigenemia would not be a useful diagnostic tool.

Environmental Impact of Tributyltin-Resistant Marine Bacteria in the Indigenous Microbial Population of Tributyltin-Polluted Surface Sediments.

PubMed

Mimura, Haruo; Yagi, Masahiro; Yoshida, Kazutoshi

2017-01-01

　We compared the TBT-resistant ability of resting cells prepared from isolates that formed colonies on nutrient agar plates containing 100 µM tributyltin (TBT) chloride, such as Photobacterium sp. TKY1, Halomonas sp. TKY2, and Photobacterium sp. NGY1, with those from taxonomically similar type strains. Photobacterium sp. TKY1 showed the highest ability among those three isolates. The number of surviving Photobacterium sp. TKY1 cells was hardly decreased after 1 h of exposure to 100 µM TBTCl, regardless of the number of resting cells in the range from 10 9.4 to 10 4.2 CFU mL -1 . In such an experimental condition, the maximum number of TBT molecules available to associate with a single cell was estimated to be approximately 6.0 x 10 11.8 . Resting cells prepared from type strains Photobacterium ganghwense JCM 12487 T and P. halotolerans LMG 22194 T , which have 16S rDNA sequences highly homologous with those of Photobacterium sp. TKY1, showed sensitivity to TBT, indicating that TBT-resistant marine bacterial species are not closely related in spite of their taxonomic similarity. We also estimated the impact of TBT-resistant bacterial species to indigenous microbial populations of TBT-polluted surface sediments. The number of surviving TBT-sensitive Vibrio natriegens ATCC 14048 T cells, 10 6.2±0.3 CFU mL -1 , was reduced to 10 4.4±0.4 CFU mL -1 when TBT-resistant Photobacterium sp. TKY1 cells, 10 9.1±0.2 CFU mL -1 , coexisted with 10 9.4±0.2 CFU mL -1 of V. natriegens ATCC 14048 T cells in the presence of 100 µM TBTCl. These results indicate that the toxicity of TBT to TBT-sensitive marine bacterial populations might be enhanced when a TBT-resistant marine bacterial species inhabits TBT-polluted surface sediments.

Rapid detection of Salmonella in milk by electrochemical magneto-immunosensing.

PubMed

Liébana, Susana; Lermo, Anabel; Campoy, Susana; Cortés, María Pilar; Alegret, Salvador; Pividori, María Isabel

2009-10-15

A very simple and rapid method for the detection of Salmonella in milk is reported. In this approach, the bacteria are captured and preconcentrated from milk samples with magnetic beads through an immunological reaction. A second polyclonal antibody labeled with peroxidase is used as serological confirmation with electrochemical detection based on a magneto-electrode. The 'IMS/m-GEC electrochemical immunosensing' approach shows a limit of detection of 5 x 10(3) and 7.5 x 10(3)CFU mL(-1) in LB and in milk diluted 1/10 in LB broth, respectively, in 50 min without any pretreatment. If the skimmed-milk is preenriched for 6h, the method is able to detect as low as 1.4 CFU mL(-1), while if it is preenriched for 8h, as low as 0.108 x CFU mL(-1) (2.7 x CFU in 25 g of milk, in 5 samples of 5 mL) are detected accordingly with the legislation. Moreover, the method is able to clearly distinguish between food pathogenic bacteria such as Salmonella and Escherichia coli. The features of this approach are discussed and compared with classical culture methods.

Inactivation of Escherichia coli by citral.

PubMed

Somolinos, M; García, D; Condón, S; Mackey, B; Pagán, R

2010-06-01

The aim was to evaluate (i) the resistance of Escherichia coli BJ4 to citral in a buffer system as a function of citral concentration, treatment medium pH, storage time and initial inoculum size, (ii) the role of the sigma factor RpoS on citral resistance of E. coli, (iii) the role of the cell envelope damage in the mechanism of microbial inactivation by citral and (iiii) possible synergistic effects of mild heat treatment and pulsed electric fields (PEF) treatment combined with citral. The initial inoculum size greatly affected the efficacy of citral against E. coli cells. Exposure to 200 microl l(-1) of citral at pH 4.0 for 24 h at 20 degrees C caused the inactivation of more than 5 log(10) cycles of cells starting at an inoculum size of 10(6) or 10(7) CFU ml(-1), whereas increasing the cell concentration to 10(9) CFU ml(-1) caused <1 log(10) cycle of inactivation. Escherichia coli showed higher resistance to citral at pH 4.0 than pH 7.0. The rpoS null mutant strain E. coli BJ4L1 was less resistant to citral than the wild-type strain. Occurrence of sublethal injury to both the cytoplasmic and outer membranes was demonstrated by adding sodium chloride or bile salts to the recovery media. The majority of sublethally injured cells by citral required energy and lipid synthesis for repair. A strongly synergistic lethal effect was shown by mild heat treatment combined with citral but the presence of citral during the application of a PEF treatment did not show any advantage. This work confirms that cell envelope damage is an important event in citral inactivation of bacteria, and it describes the key factors on the inactivation of E. coli cells by citral. Knowledge about the mechanism of microbial inactivation by citral helps establish successful combined preservation treatments.

Inhibition of Listeria monocytogenes by propionic acid-based ingredients in cured deli-style Turkey.

PubMed

Glass, Kathleen A; McDonnell, Lindsey M; Von Tayson, Roxanne; Wanless, Brandon; Badvela, Mani

2013-12-01

Listeria monocytogenes growth can be controlled on ready-to-eat meats through the incorporation of antimicrobial ingredients into the formulation or by postlethality kill steps. However, alternate approaches are needed to provide options that reduce sodium content but maintain protection against pathogen growth in meats after slicing. The objective of this study was to determine the inhibition of L. monocytogenes by propionic acid-based ingredients in high-moisture, cured turkey stored at 4 or 7°C. Six formulations of sliced, cured (120 ppm of NaNO2 ), deli-style turkey were tested, including control without antimicrobials, 3.2% lactate-diacetate blend (LD), 0.4% of a liquid propionate-benzoate-containing ingredient, or 0.3, 0.4, and 0.5% of a liquid propionate-containing ingredient. Products were inoculated with 5 log CFU L. monocytogenes per 100-g package (3 log CFU/ml rinsate), vacuum-sealed, and stored at 4 or 7°C for up to 12 weeks; and populations were enumerated by plating on modified Oxford agar. As expected, the control without antimicrobials supported rapid growth, with >2 log average per ml rinsate increase within 4 weeks of storage at 4°C, whereas growth was observed at 6 weeks for the LD treatment. For both replicate trials, all treatments that contained liquid propionate or propionate-benzoate limited L. monocytogenes growth to an increase of <1 log through 9 weeks storage at 4°C. Sporadic growth (>1-log increase) was observed in individual samples for all propionate-containing treatments at weeks 10, 11, and 12. As expected, L. monocytogenes grew more rapidly when products were stored at 7°C, but trends in relative inhibition were similar to those observed at 4°C. These results verify that propionate-based ingredients inhibit growth of L. monocytogenes on sliced, high-moisture, cured turkey and can be considered as an alternative to reduce sodium-based salts while maintaining food safety.

Differences in biofilm formation of produce and poultry Salmonella enterica isolates and their persistence on spinach plants

USDA-ARS?s Scientific Manuscript database

Repeat irrigation of spinach plants with water containing Salmonella was used to determine Salmonella persistence on spinach leaves. Spinach plants were irrigated four times (biweekly) with water containing ca. 2.1 log CFU Salmonella per 100 ml water (the maximum generic E. coli MPN recommended by...

Physical and antimicrobial properties of anise oil loaded nanoemulsions on the survival of foodborne pathogens.

PubMed

Topuz, Osman Kadir; Özvural, Emin Burçin; Zhao, Qin; Huang, Qingrong; Chikindas, Michael; Gölükçü, Muharrem

2016-07-15

The purpose of this research was to investigate antimicrobial effects of nano emulsions of anise oil (AO) on the survival of common food borne pathogens, Listeria monocytogenes and Escherichia coli O157:H7. Series of emulsions containing different level of anise oil as potential antimicrobial delivery systems were prepared. Antimicrobial activities of bulk anise oil and its emulsions (coarse and nano) was tested by the minimum inhibitory concentration and time kill assay. Our results showed that bulk anise oil reduced the population of E. coli O157:H7 and L. monocytogenes by 1.48 and 0.47 log cfu/ml respectively after 6 h of contact time. However, under the same condition anise oil nanoemulsion (AO75) reduced E. coli O157:H7 and L. monocytogenes count by 2.51 and 1.64 log cfu/ml, respectively. Physicochemical and microbial analyses indicated that both nano and coarse emulsions of anise oil showed better and long-term physicochemical stability and antimicrobial activity compared to bulk anise oil. Copyright © 2016 Elsevier Ltd. All rights reserved.

Changes in microbial composition and the prevalence of foodborne pathogens in crab marinated in soy sauce produced by six manufacturing plants.

PubMed

Kim, Sun Ae; Choi, Eun Sook; Kim, Nam Hee; Kim, Hye Won; Lee, Na Young; Cho, Tae Jin; Jo, Jun Il; Kim, Soon Han; Lee, Soon Ho; Ha, Sang Do; Rhee, Min Suk

2017-04-01

The present study examined the changes in microbiological composition during the production process of crab marinated in soy sauce, potential microbial hazards, potential contamination routes and effective critical control points. Crab and soy sauce samples were obtained from six different manufacturing plants at different stages, and their microbiological content was comprehensively assessed by quantitative and qualitative analyses. The results revealed the following: (1) the final products contained 4.0 log colony-forming units (CFU) g -1 aerobic plate counts (APCs) and 1.1 log CFU g -1 coliforms, which may have been introduced from the raw materials (the level of APCs in raw crab and soy sauce mixed with other ingredients was 3.8 log CFU g -1 and 4.0 log CFU mL -1 respectively); (2) marination of crab in soy sauce may allow cross-contamination by coliforms; (3) only Bacillus cereus and Staphylococcus aureus were qualitatively detected in samples at different stages of manufacture (detection rate of 28 and 5.6% respectively), and these bacteria may impact the microbiological quality and safety of crab marinated in soy sauce; and (4) bacterial counts were either maintained or increased during the manufacturing process (suggesting that no particular step can be targeted to reduce bacterial counts). Proper management of raw materials and the marination process are effective critical control points, and alternative interventions may be needed to control bacterial quantity. The results provide important basic information about the production of crab marinated in soy sauce and may facilitate effective implementation of sanitary management practices in related industries and research fields. © 2016 Society of Chemical Industry. © 2016 Society of Chemical Industry.

Diazonium-based impedimetric aptasensor for the rapid label-free detection of Salmonella typhimurium in food sample.

PubMed

Bagheryan, Zahra; Raoof, Jahan-Bakhsh; Golabi, Mohsen; Turner, Anthony P F; Beni, Valerio

2016-06-15

Fast and accurate detection of microorganisms is of key importance in clinical analysis and in food and water quality monitoring. Salmonella typhimurium is responsible for about a third of all cases of foodborne diseases and consequently, its fast detection is of great importance for ensuring the safety of foodstuffs. We report the development of a label-free impedimetric aptamer-based biosensor for S. typhimurium detection. The aptamer biosensor was fabricated by grafting a diazonium-supporting layer onto screen-printed carbon electrodes (SPEs), via electrochemical or chemical approaches, followed by chemical immobilisation of aminated-aptamer. FTIR-ATR, contact angle and electrochemical measurements were used to monitor the fabrication process. Results showed that electrochemical immobilisation of the diazonium-grafting layer allowed the formation of a denser aptamer layer, which resulted in higher sensitivity. The developed aptamer-biosensor responded linearly, on a logarithm scale, over the concentration range 1 × 10(1) to 1 × 10(8)CFU mL(-1), with a limit of quantification (LOQ) of 1 × 10(1) CFU mL(-1) and a limit of detection (LOD) of 6 CFU mL(-1). Selectivity studies showed that the aptamer biosensor could discriminate S. typhimurium from 6 other model bacteria strains. Finally, recovery studies demonstrated its suitability for the detection of S. typhimurium in spiked (1 × 10(2), 1 × 10(4) and 1 × 10(6) CFU mL(-1)) apple juice samples. Copyright © 2016 Elsevier B.V. All rights reserved.

Simulated-use validation of a sponge ATP method for determining the adequacy of manual cleaning of endoscope channels.

PubMed

Alfa, Michelle J; Olson, Nancy

2016-05-04

The objective of this study was to validate the relative light unit (RLU) cut-off of adequate cleaning of flexible colonoscopes for an ATP (adenosine tri-phosphate) test kit that used a sponge channel collection method. This was a simulated-use study. The instrument channel segment of a flexible colonoscope was soiled with ATS (artificial test soil) containing approximately 8 Log10 Enterococcus faecalis and Pseudomonas aeruginosa/mL. Full cleaning, partial cleaning and no cleaning were evaluated for ATP, protein and bacterial residuals. Channel samples were collected using a sponge device to assess residual RLUs. Parallel colonoscopes inoculated and cleaned in the same manner were sampled using the flush method to quantitatively assess protein and bacterial residuals. The protein and viable count benchmarks for adequate cleaning were <6.4 ug/cm(2) and <4 Log10 cfu/cm(2). The negative controls for the instrument channel, over the course of the study remained low with on average 14 RLUs, 0.04 ug/cm(2) protein and 0.025 Log10 cfu/cm(2). Partial cleaning resulted in an average of 6601 RLUs, 3.99 ug/cm(2), 5.25 Log10 cfu/cm(2) E. faecalis and 4.48 Log10 cfu/cm(2) P. aeruginosa. After full cleaning, the average RLU was 29 (range 7-71 RLUs) and the average protein, E. faecalis and P. aeruginosa residuals were 0.23 ug/cm(2), 0.79 and 1.61 Log10 cfu/cm(2), respectively. The validated cut-off for acceptable manual cleaning was set at ≤100 RLUs for the sponge collected channel ATP test kit.

Development of TiO2 powder-coated food packaging film and its ability to inactivate Escherichia coli in vitro and in actual tests.

PubMed

Chawengkijwanich, Chamorn; Hayata, Yasuyoshi

2008-04-30

Titanium dioxide (TiO2) has attracted a great deal of attention as a photocatalytic disinfecting material in the food and environmental industry. TiO2 has been used to inactivate a wide variety of microorganisms in many applications. In the present study, we aimed to develop a TiO2 powder-coated packaging film and clarify its ability to inactivate Escherichia coli both in vitro and in actual tests, using two different particle sizes and two types of illumination at different intensities. No inhibition effect of the testing method itself on the growth of E. coli was observed. The cells of E. coli were found to have decreased 3 log CFU/ml after 180 min of illumination by two 20 W black-light bulbs (wavelength of 300-400 nm) on TiO2-coated oriented-polypropylene (OPP) film, while E. coli decreased 1 log CFU/m with black-light illumination of uncoated OPP film. The results showed that both ultraviolet A (UVA; wavelength of 315-400 nm) alone and TiO2-coated OPP film combined with UVA reduced the number of E. coli cell in vitro, but that the reduction of E. coli cell numbers was greater by TiO2-coated OPP film combined with UVA. The antimicrobial effect of TiO2-coated film is dependent on the UVA light intensity (0, <0.05 and 1 mW/cm2) and the kind of artificial light (black-light and daylight fluorescent bulbs), but it is independent of the particle size of TiO2 coating on the surface of OPP film. The surviving cell numbers of E. coli on TiO2-coated film decreased 3 log and 0.35 log CFU/ml after 180 min of illumination by two 20 W black bulbs and two 20 W daylight fluorescent bulbs, respectively. Despite the lesser efficacy of the photocatalytic method with fluorescent lights, the survival of E. coli cells using this method was 50% of that using fluorescent lights alone. In the actual test, the number of E. coli cells from cut lettuce stored in a TiO2-coated film bag irradiated with UVA light decreased from 6.4 on Day 0 to 4.9 log CFU/g on Day 1, while that of an uncoated film bag irradiated with UVA light decreased from 6.4 to 6.1 log CFU/g after 1 day of storage. The result shows that the TiO2-coated film could reduce the microbial contamination on the surface of solid food products and thus reduce the risks of microbial growth on fresh-cut produce.

The effect of stable bedding materials on dust levels, microbial air contamination and equine respiratory health.

PubMed

Kwiatkowska-Stenzel, Agnieszka; Witkowska, Dorota; Sowińska, Janina; Stopyra, Artur

2017-12-01

The choice of bedding material affects the quality of air in a stable and, consequently, the respiratory health of horses and humans. The risk of respiratory problems can be mitigated by improving the quality of air in the stable. The choice of bedding material is particularly important in cold climate conditions where horses are kept indoors throughout the year. This study examined the impact of three bedding materials: straw (S), peat with shavings (PS), and crushed wood pellets (CWP). The investigated factors were air contamination, including dust contamination and microbial (bacterial and fungal) contamination, and the condition of the equine respiratory tract. The condition of the respiratory tract was evaluated based on the results of arterial blood biochemistry tests and endoscopic evaluations of the upper respiratory tract. Mechanical dust contamination was lowest for PS (1.09mg/m 3 ) and highest for CWP (4.07mg/m 3 ). Bacterial contamination (in CFU - colony forming units) was highest for PS (5.14log 10 CFU/m 3 ) and lowest for CWP (4.81log 10 CFU/m 3 ). Fungal air contamination was lowest for CWP (4.54log 10 CFU/m 3 ) and highest for S (4.82log 10 CFU/m 3 ) and PS (4.88log 10 CFU/m 3 ). An analysis of physiological indicators revealed that all horses were clinically healthy regardless of the type of applied bedding. The type of bedding material did not exert a clear influence on arterial blood biochemistry or the results of endoscopic evaluations of the respiratory tract; however, the use of alternative for straw bedding materials improved endoscopy results. Copyright © 2017 Elsevier Ltd. All rights reserved.

Hartmannella vermiformis inhibition of Legionella pneumophila cultivability.

PubMed

Buse, Helen Y; Donohue, Maura J; Ashbolt, Nicholas J

2013-10-01

Hartmannella vermiformis and Acanthamoeba polyphaga are frequently isolated from drinking water and permissive to Legionella pneumophila parasitization. In this study, extracellular factor(s) produced by H. vermiformis and A. polyphaga were assessed for their effects on cultivability of L. pneumophila. Page's amoeba saline (PAS) was used as an encystment medium for H. vermiformis and A. polyphaga monolayers, and the culture supernatants (HvS and ApS, respectively) were assessed against L. pneumophila growth. Compared to PAS and ApS, HvS significantly inhibited L. pneumophila strain Philadelphia-1 (Ph-1) cultivability by 3 log(10) colony forming unit (CFU) mL(-1) after 3 days of exposure compared to <0.5 log(10) CFU mL(-1) reduction of strain Lp02 (P < 0.001). Flow cytometric analysis revealed changes in the percentage and cultivability of three bacterial subpopulations: intact/slightly damaged membrane (ISM), undefined membrane status (UD), and mixed type (MT). After 3 days of HvS exposure, the MT subpopulation decreased significantly (31.6 vs 67.2 %, respectively, P < 0.001), while the ISM and UD subpopulations increased (+26.7 and +6.9 %, respectively) with the ISM subpopulation appearing as viable but nonculturable (VBNC) cells. HvS was separated into two fractions based on molecular weight, with more than 99 % of the L. pneumophila inhibition arising from the <5 kDa fraction (P < 0.001). Liquid chromatography indicated the inhibitory molecule(s) are likely polar and elute from a Novapak C18 column between 6 and 15 min. These results demonstrate that H. vermiformis is capable of extracellular modulation of L. pneumophila cultivability and probably promote the VBNC state for this bacterium.

Specificity of coliphages in evaluating marker efficacy: a new insight for water quality indicators.

PubMed

Mookerjee, Subham; Batabyal, Prasenjit; Halder, Madhumanti; Palit, Anup

2014-11-01

Conventional procedures for qualitative assessment of coliphage are time consuming multiple step approach for achieving results. A modified and rapid technique has been introduced for determination of coliphage contamination among potable water sources during water borne outbreaks. During December 2013, 40 water samples from different potable water sources, were received for water quality analyses, from a jaundice affected Municipality of West Bengal, India. Altogether, 30% water samples were contaminated with coliform (1-20 cfu/ml) and 5% with E. coli (2-5 cfu/ml). Among post-outbreak samples, preponderance of coliform has decreased (1-4 cfu/ml) with total absence of E. coli. While standard technique has detected 55% outbreak samples with coliphage contamination, modified technique revealed that 80%, double than that of bacteriological identification rate, were contaminated with coliphages (4-20 pfu/10 ml). However, post-outbreak samples were detected with 1-5 pfu/10 ml coliphages among 20% samples. Coliphage detection rate through modified technique was nearly double (50%) than that of standard technique (27.5%). In few samples (with coliform load of 10-100 cfu/ml), while modified technique could detect coliphages among six samples (10-20 pfu/10 ml), standard protocol failed to detect coliphage in any of them. An easy, rapid and accurate modified technique has thereby been implemented for coliphage assessment from water samples. Coliform free water does not always signify pathogen free potable water and it is demonstrated that coliphage is a more reliable 'biomarker' to ascertain contamination level in potable water. Copyright © 2014 Elsevier B.V. All rights reserved.

Changes in the Microbial Composition of Microbrewed Beer during the Process in the Actual Manufacturing Line.

PubMed

Kim, S A; Jeon, S H; Kim, N H; Kim, H W; Lee, N Y; Cho, T J; Jung, Y M; Lee, S H; Hwang, I G; Rhee, M S

2015-12-01

This study investigated changes in the microbial composition of microbrewed beer during the manufacturing processes and identified potential microbial hazards, effective critical quality control points, and potential contamination routes. Comprehensive quantitative (aerobic plate count, lactic acid bacteria, fungi, acetic acid bacteria, coliforms, and Bacillus cereus) and qualitative (Escherichia coli and eight foodborne pathogens) microbiological analyses were performed using samples of raw materials (malt and manufacturing water), semiprocessed products (saccharified wort, boiled wort, and samples taken during the fermentation and maturation process), and the final product obtained from three plants. The initial aerobic plate count and lactic acid bacteria counts in malt were 5.2 and 4.3 log CFU/g, respectively. These counts were reduced to undetectable levels by boiling but were present at 2.9 and 0.9 log CFU/ml in the final product. Fungi were initially present at 3.6 log CFU/g, although again, the microbes were eliminated by boiling; however, the level in the final product was 4.6 log CFU/ml. No E. coli or foodborne pathogens (except B. cereus) were detected. B. cereus was detected at all stages, although it was not present in the water or boiled wort (total detection rate ¼ 16.4%). Results suggest that boiling of the wort is an effective microbial control measure, but careful management of raw materials and implementation of effective control measures after boiling are needed to prevent contamination of the product after the boiling step. The results of this study may constitute useful and comprehensive information regarding the microbiological quality of microbrewed beer.

Risk evaluation for staphylococcal food poisoning in processed milk produced with skim milk powder.

PubMed

Soejima, T; Nagao, E; Yano, Y; Yamagata, H; Kagi, H; Shinagawa, K

2007-04-01

The growth of S. aureus and the production of staphylococcal enterotoxin A (SEA) in skim milk concentrates stored at inappropriate temperatures in a recovery milk tank (tank for excess concentrated skim milk) used in the manufacture of skimmed milk powder were investigated. Also, it was estimated if a possible outbreak of food poisoning would occur if the contaminated skimmed milk powder was used in the manufacture of processed milk. Skim milk concentrates with milk solid content of 15, 25, and 35% were inoculated with S. aureus at 1-2 log CFU/ml and incubated at 15, 25, or 35 degrees C for 0 to 24 h with or without shaking. Bacterial growth and the level of SEA production were measured. At 35 degrees C with shaking, there was a significant difference (p<0.05) in one way layout analysis of variance, and it was demonstrated that the growth of S. aureus and SEA production could be milk solid content-dependent. Shaking accelerated the growth of S. aureus and SEA production at 35 degrees C. Generally, skim milk powder is produced by mixing a set percentage of skim milk concentrates (recovery milk) from the recovery milk tank into raw milk. If recovery milk contaminated with S. aureus at levels of 1-2 log CFU/ml is kept at 15 to 35 degrees C due to a power failure, it was estimated that processed milk consumption of 670-1200 ml, 420-1500 ml and 18-83 ml would trigger the onset of food poisoning symptoms when skim milk concentrates (recovery milk) are stored at 25 degrees C for 24 h, 35 degrees C for 10 h, and 35 degrees C for 24 h, respectively, during the production of the skim milk powder. Based on these consumption levels, it was concluded that, if recovery milk cannot be refrigerated and is stored at room temperature (25 to 35 degrees C), it must be used within 8 h and preferably within 6 h.

Growth of Listeria monocytogenes, Salmonella spp., Escherichia coli O157:H7, and Staphylococcus aureus on cheese during extended storage at 25°C.

PubMed

Leong, Wan Mei; Geier, Renae; Engstrom, Sarah; Ingham, Steve; Ingham, Barbara; Smukowski, Marianne

2014-08-01

Potentially hazardous foods require time/temperature control for safety. According to the U.S. Food and Drug Administration Food Code, most cheeses are potentially hazardous foods based on pH and water activity, and a product assessment is required to evaluate safety of storage >6 h at 21°C. We tested the ability of 67 market cheeses to support growth of Listeria monocytogenes (LM), Salmonella spp. (SALM), Escherichia coli O157:H7 (EC), and Staphylococcus aureus (SA) over 15 days at 25°C. Hard (Asiago and Cheddar), semi-hard (Colby and Havarti), and soft cheeses (mozzarella and Mexican-style), and reduced-sodium or reduced-fat types were tested. Single-pathogen cocktails were prepared and individually inoculated onto cheese slices (∼10(5) CFU/g). Cocktails were 10 strains of L. monocytogenes, 6 of Salmonella spp., or 5 of E. coli O157:H7 or S. aureus. Inoculated slices were vacuum packaged and stored at 25°C for ≤ 15 days, with surviving inocula enumerated every 3 days. Percent salt-in-the-moisture phase, percent titratable acidity, pH, water activity, and levels of indigenous/starter bacteria were measured. Pathogens did not grow on 53 cheeses, while 14 cheeses supported growth of SA, 6 of SALM, 4 of LM, and 3 of EC. Of the cheeses supporting pathogen growth, all supported growth of SA, ranging from 0.57 to 3.08 log CFU/g (average 1.70 log CFU/g). Growth of SALM, LM, and EC ranged from 1.01 to 3.02 log CFU/g (average 2.05 log CFU/g), 0.60 to 2.68 log CFU/g (average 1.60 log CFU/g), and 0.41 to 2.90 log CFU/g (average 1.69 log CFU/g), respectively. Pathogen growth varied within cheese types or lots. Pathogen growth was influenced by pH and percent salt-in-the-moisture phase, and these two factors were used to establish growth/no-growth boundary conditions for safe, extended storage (≤25°C) of pasteurized milk cheeses. Pathogen growth/no-growth could not be predicted for Swiss-style cheeses, mold-ripened or bacterial surface-ripened cheeses, and cheeses made with nonbovine milk, as insufficient data were gathered. This challenge study data can support science-based decision making in a regulatory framework.

Inoculum Effect on the Efficacies of Amoxicillin-Clavulanate, Piperacillin-Tazobactam, and Imipenem against Extended-Spectrum β-Lactamase (ESBL)-Producing and Non-ESBL-Producing Escherichia coli in an Experimental Murine Sepsis Model

PubMed Central

López-Cerero, L.; López-Rojas, R.; Egea, P.; Domínguez-Herrera, J.; Rodríguez-Baño, J.; Pascual, A.; Pachón, J.

2013-01-01

Escherichia coli is commonly involved in infections with a heavy bacterial burden. Piperacillin-tazobactam and carbapenems are among the recommended empirical treatments for health care-associated complicated intra-abdominal infections. In contrast to amoxicillin-clavulanate, both have reduced in vitro activity in the presence of high concentrations of extended-spectrum β-lactamase (ESBL)-producing and non-ESBL-producing E. coli bacteria. Our goal was to compare the efficacy of these antimicrobials against different concentrations of two clinical E. coli strains, one an ESBL-producer and the other a non-ESBL-producer, in a murine sepsis model. An experimental sepsis model {∼5.5 log10 CFU/g [low inoculum concentration (LI)] or ∼7.5 log10 CFU/g [high inoculum concentration (HI)]} using E. coli strains ATCC 25922 (non-ESBL producer) and Ec1062 (CTX-M-14 producer), which are susceptible to the three antimicrobials, was used. Amoxicillin-clavulanate (50/12.5 mg/kg given intramuscularly [i.m.]), piperacillin-tazobactam (25/3.125 mg/kg given intraperitoneally [i.p.]), and imipenem (30 mg/kg i.m.) were used. Piperacillin-tazobactam and imipenem reduced spleen ATCC 25922 strain concentrations (−2.53 and −2.14 log10 CFU/g [P < 0.05, respectively]) in the HI versus LI groups, while amoxicillin-clavulanate maintained its efficacy (−1.01 log10 CFU/g [no statistically significant difference]). Regarding the Ec1062 strain, the antimicrobials showed lower efficacy in the HI than in the LI groups: −0.73, −1.89, and −1.62 log10 CFU/g (P < 0.05, for piperacillin-tazobactam, imipenem, and amoxicillin-clavulanate, respectively, although imipenem and amoxicillin-clavulanate were more efficacious than piperacillin-tazobactam). An adapted imipenem treatment (based on the time for which the serum drug concentration remained above the MIC obtained with a HI of the ATCC 25922 strain) improved its efficacy to −1.67 log10 CFU/g (P < 0.05). These results suggest that amoxicillin-clavulanate could be an alternative to imipenem treatment of infections caused by ESBL- and non-ESBL-producing E. coli strains in patients with therapeutic failure with piperacillin-tazobactam. PMID:23439636

Inoculum effect on the efficacies of amoxicillin-clavulanate, piperacillin-tazobactam, and imipenem against extended-spectrum β-lactamase (ESBL)-producing and non-ESBL-producing Escherichia coli in an experimental murine sepsis model.

PubMed

Docobo-Pérez, F; López-Cerero, L; López-Rojas, R; Egea, P; Domínguez-Herrera, J; Rodríguez-Baño, J; Pascual, A; Pachón, J

2013-05-01

Escherichia coli is commonly involved in infections with a heavy bacterial burden. Piperacillin-tazobactam and carbapenems are among the recommended empirical treatments for health care-associated complicated intra-abdominal infections. In contrast to amoxicillin-clavulanate, both have reduced in vitro activity in the presence of high concentrations of extended-spectrum β-lactamase (ESBL)-producing and non-ESBL-producing E. coli bacteria. Our goal was to compare the efficacy of these antimicrobials against different concentrations of two clinical E. coli strains, one an ESBL-producer and the other a non-ESBL-producer, in a murine sepsis model. An experimental sepsis model {~5.5 log10 CFU/g [low inoculum concentration (LI)] or ~7.5 log(10) CFU/g [high inoculum concentration (HI)]} using E. coli strains ATCC 25922 (non-ESBL producer) and Ec1062 (CTX-M-14 producer), which are susceptible to the three antimicrobials, was used. Amoxicillin-clavulanate (50/12.5 mg/kg given intramuscularly [i.m.]), piperacillin-tazobactam (25/3.125 mg/kg given intraperitoneally [i.p.]), and imipenem (30 mg/kg i.m.) were used. Piperacillin-tazobactam and imipenem reduced spleen ATCC 25922 strain concentrations (-2.53 and -2.14 log10 CFU/g [P < 0.05, respectively]) in the HI versus LI groups, while amoxicillin-clavulanate maintained its efficacy (-1.01 log10 CFU/g [no statistically significant difference]). Regarding the Ec1062 strain, the antimicrobials showed lower efficacy in the HI than in the LI groups: -0.73, -1.89, and -1.62 log10 CFU/g (P < 0.05, for piperacillin-tazobactam, imipenem, and amoxicillin-clavulanate, respectively, although imipenem and amoxicillin-clavulanate were more efficacious than piperacillin-tazobactam). An adapted imipenem treatment (based on the time for which the serum drug concentration remained above the MIC obtained with a HI of the ATCC 25922 strain) improved its efficacy to -1.67 log10 CFU/g (P < 0.05). These results suggest that amoxicillin-clavulanate could be an alternative to imipenem treatment of infections caused by ESBL- and non-ESBL-producing E. coli strains in patients with therapeutic failure with piperacillin-tazobactam.

Efficacy of neutral electrolyzed water to inactivate Escherichia coli, Listeria monocytogenes, Pseudomonas aeruginosa, and Staphylococcus aureus on plastic and wooden kitchen cutting boards.

PubMed

Deza, M A; Araujo, M; Garrido, M J

2007-01-01

This study evaluated the efficacy of neutral electrolyzed water (NEW; 64.1 mg/liter of active chlorine) to reduce populations of Escherichia coli, Pseudomonas aeruginosa, Staphylococcus aureus, and Listeria monocytogenes on plastic and wooden kitchen cutting boards. Its effectiveness was compared with that of a sodium hypochlorite solution (NaClO; 62.3 mg/liter of active chlorine). Inoculated portions of cutting boards were rinsed in either NEW or NaClO solutions, or deionized water (control). Plastic boards were rinsed for 1 min and wooden boards for 1 and 5 min. After each treatment, the surviving population of each strain was determined on the surface and in the soaking water. No significant difference (P > or = 0.05) was found between the final populations of each strain with regard to the treatment solutions (NEW or NaClO). However, a significant difference (P < or = 0.05) was revealed between surface materials after 1 min of washing. Whereas in plastic boards the initial bacterial populations were reduced by 5 log CFU/50 cm2, in wooden cutting boards they underwent a reduction of <3 log CFU/50 cm2. A 5-min exposure time yielded reductions of about 4 log CFU/50 cm2. The surviving populations of all bacteria in NEW and NaCIO washing solutions were <1 log CFU/ml after soaking both surfaces. This study revealed that NEW treatment is an effective method for reducing microbial contamination on plastic and wooden cutting boards. NEW efficacy was comparable to that of NaCIO, with the advantage of having a larger storage time.

Female urinary tract infection in primary health care: bacteriological and clinical characteristics.

PubMed

Osterberg, E; Hallander, H O; Kallner, A; Lundin, A; Svensson, S B; Aberg, H

1990-01-01

Female patients with symptoms of urinary tract infection (n = 1136) were studied in primary health care with respect to (a) clinical symptoms as predictors of bacteriuria; (b) relation between aetiological agent and clinical picture, especially for P-fimbriated Escherichia coli; and (c) clinical findings in cases with 10(2)- less than 10(5) CFU/ml of E. coli. Prevalence of bacteriuria (greater than or equal to 10(5) CFU/ml) was 61%. Concurrence of urgency/frequency and dysuria, short duration of symptoms and hematuria increased the probability of bacteriuria and were also significantly more frequent among cases with low counts of E. coli (10(2) less than 10(5) CFU/ml in pure culture or mixed flora) than among cases with sterile urine, indicating an aetiological role of E. coli in many of those cases. Infections with P-fimbriated E. coli were as benign as the P-fimbriae-negative. The rate of P-fimbriation was 29% in specimens containing greater than or equal to 10(5) CFU/ml of E. coli, 30% among specimens with less than 10(5) CFU/ml in pure culture and 10% in specimens containing less than 10(5) CFU/ml of E. coli in mixed culture. Patients infected with Klebsiella, Enterobacter or Proteus did not show a higher rate of previous urinary tract disease or anomalies.

Analyzing indicator microorganisms, antibiotic resistant Escherichia coli, and regrowth potential of foodborne pathogens in various organic fertilizers.

PubMed

Miller, Cortney; Heringa, Spencer; Kim, Jinkyung; Jiang, Xiuping

2013-06-01

This study analyzed various organic fertilizers for indicator microorganisms, pathogens, and antibiotic-resistant Escherichia coli, and evaluated the growth potential of E. coli O157:H7 and Salmonella in fertilizers. A microbiological survey was conducted on 103 organic fertilizers from across the United States. Moisture content ranged from approximately 1% to 86.4%, and the average pH was 7.77. The total aerobic mesophiles ranged from approximately 3 to 9 log colony-forming units (CFU)/g. Enterobacteriaceae populations were in the range of <1 to approximately 7 log CFU/g, while coliform levels varied from <1 to approximately 6 log CFU/g. Thirty samples (29%) were positive for E. coli, with levels reaching approximately 6 log CFU/g. There were no confirmed positives for E. coli O157:H7, Salmonella, or Listeria monocytogenes. The majority of E. coli isolates (n=73), confirmed by glutamate decarboxylase (gad) PCR, were from group B1 (48%) and group A (32%). Resistance to 16 antibiotics was examined for 73 E. coli isolates, with 11 isolates having resistance to at least one antibiotic, 5 isolates to ≥ 2 antibiotics, and 2 isolates to ≥ 10 antibiotics. In the presence of high levels of background aerobic mesophiles, Salmonella and E. coli O157:H7 grew approximately 1 log CFU/g within 1 day of incubation in plant-based compost and fish emulsion-based compost, respectively. With low levels of background aerobic mesophiles, Salmonella grew approximately 2.6, 3.0, 3.0, and 3.2 log CFU/g in blood, bone, and feather meals and the mixed-source fertilizer, respectively, whereas E. coli O157:H7 grew approximately 4.6, 4.0, 4.0, and 4.8 log CFU/g, respectively. Our results revealed that the microbiological quality of organic fertilizers varies greatly, with some fertilizers containing antibiotic resistant E. coli and a few supporting the growth of foodborne pathogens after reintroduction into the fertilizer.

Microbiological quality of fresh, minimally-processed fruit and vegetables, and sprouts from retail establishments.

PubMed

Abadias, M; Usall, J; Anguera, M; Solsona, C; Viñas, I

2008-03-31

A survey of fresh and minimally-processed fruit and vegetables, and sprouts was conducted in several retail establishments in the Lleida area (Catalonia, Spain) during 2005-2006 to determine whether microbial contamination, and in particular potentially pathogenic bacteria, was present under these commodities. A total of 300 samples--including 21 ready-to-eat fruits, 28 whole fresh vegetables, 15 sprout samples and 237 ready-to-eat salads containing from one to six vegetables--were purchased from 4 supermarkets. They were tested for mesophilic and psychrotrophic aerobic counts, yeasts and moulds, lactic acid bacteria, Enterobacteriaceae, presumptive E. coli and Listeria monocytogenes counts as well as for the presence of Salmonella, E. coli O157:H7, Yersinia enterocolitica and thermotolerant Campylobacter. Results for the fresh-cut vegetables that we analyzed showed that, in general, the highest microorganism counts were associated with grated carrot, arugula and spinach (7.8, 7.5 and 7.4 log cfu g(-1) of aerobic mesophilic microorganisms; 6.1, 5.8 and 5.2 log cfu g(-1) of yeast and moulds; 5.9, 4.0 and 5.1 log cfu g(-1) lactic acid bacteria and 6.2, 5.3 and 6.0 log cfu g(-1) of Enterobacteriaceae). The lowest counts were generally associated with fresh-cut endive and lettuce (6.2 and 6.3 log cfu g(-1) of aerobic mesophilic microorganisms; 4.4 and 4.6 log cfu g(-1) of yeast and moulds; 2.7 and 3.8 log cfu g(-1) lactic acid bacteria and 4.8 and 4.4 log cfu g(-1) of Enterobacteriaceae). Counts of psychrotrophic microorganisms were as high as those of mesophilic microorganisms. Microbiological counts for fresh-cut fruit were very low. Sprouts were highly contaminated with mesophilic (7.9 log cfu g(-1)), psychrotrophic microorganisms (7.3 log cfu g(-1)) and Enterobacteriaceae (7.2 log cfu g(-1)) and showed a high incidence of E. coli (40% of samples). Of the samples analyzed, four (1.3%) were Salmonella positive and two (0.7%) harboured L. monocytogenes. None of the samples was positive for E. coli O157:H7, pathogenic Y. enterocolitica or thermotolerant Campylobacter.

In vitro effectiveness of antimicrobial photodynamic therapy (APDT) using a 660 nm laser and malachite green dye in Staphylococcus aureus biofilms arranged on compact and cancellous bone specimens.

PubMed

Rosa, Luciano Pereira; da Silva, Francine Cristina; Nader, Sumaia Alves; Meira, Giselle Andrade; Viana, Magda Souza

2014-11-01

The aim of this study was to evaluate the in vitro effectiveness of antimicrobial photodynamic therapy (APDT) using a 660 nm visible laser combined with malachite green (MG) dye in the inactivation of Staphylococcus aureus (ATCC 25923) biofilms formed within compact and cancellous bone specimens. Specimens of 80 compact bones and 80 cancellous bones were contaminated with a standard suspension of S. aureus and incubated for 14 days at 37 °C to allow for the formation of biofilms. The specimens were divided into the following groups (n = 10) according to the treatment conditions: PS-L - (control - no treatment), PS+L - (only MG for 5 min), PS-L + 90 (only laser irradiation for 90 s), PS-L + 180 (only laser irradiation for 180 s), PS-L + 300 (only laser irradiation for 300 s), APDT90 (APDT for 90 s), APDT180 (APDT for 180 s), and APDT300 (APDT for 300 s). The findings were statistically analyzed using an ANOVA 5%. All of the experimental groups were significantly different from the control group for both the compact and cancellous bone specimens. The compact bone specimens that received APDT treatment (for either 90, 180, or 300 s) showed reductions in the log10 CFU/ml of S. aureus by a magnitude of 4 log10. Cancellous bone specimens treated with 300 s of APDT showed the highest efficacy, and these specimens had a reduction in S. aureus CFU/ml by a factor of 3 log10. APDT treatment using these proposed parameters in combination with MG was effective at inactivating S. aureus biofilms in compact and cancellous bone specimens.

Behavior of Listeria monocytogenes on Mortadella Formulated Using a Natural, Clean-Label Antimicrobial Agent during Extended Storage at 4 or 12°C.

PubMed

Porto-Fett, Anna C S; Campano, Stephen G; Rieker, Marcus; Stahler, Laura J; McGEARY, Lianna; Shane, Laura E; Shoyer, Bradley A; Osoria, Manuela; Luchansky, John B

2018-05-01

All-pork mortadella, an Italian-style deli meat, was produced by a local artisanal meat producer with or without 1.0 or 1.5% liquid buffered vinegar (LBV), 0.4, 0.6, or 1.0% dry buffered vinegar (DBV), or a 2.5% blend of potassium lactate and sodium diacetate (KLac). In each of three trials, mortadella was sliced (ca. 1.5 cm thick, ca. 30 g) and surface inoculated with 250 μL per side of a five-strain mixture of Listeria monocytogenes (ca. 3.8 log CFU per slice). The packages were vacuum sealed and then stored at 4 or 12°C. In the absence of antimicrobials, L. monocytogenes levels increased by ca. 2.6 and 6.0 log CFU per slice after up to 120 or 28 days at 4 or 12°C, respectively. With inclusion of 1.0 or 1.5% LBV, 1.0% DBV, or 2.5% KLac as ingredients, pathogen levels decreased by ca. 0.3 to 0.7 log CFU per slice after 120 days at 4°C, whereas with inclusion of 0.4 or 0.6% DBV, L. monocytogenes levels increased by ca. 1.2 and 0.8 log CFU per slice, respectively. After 28 days at 12°C, inclusion of 2.5% KLac, 1.0 or 1.5% LBV, or 0.4 or 0.6% DBV resulted in a ca. 1.4- to 5.7-log increase in L. monocytogenes levels. When 1.0% DBV was included in the formulation, pathogen levels remained unchanged after 28 days at 12°C. However, product quality was lessened at this abusive storage temperature (12°C) for all treatments by the end of storage. Thus, inclusion of LBV or DBV, as clean-label ingredients, in mortadella is equally effective as KLac for controlling L. monocytogenes during storage at 4°C without adversely affecting product quality.

Sanitizing radish seeds by simultaneous treatments with gaseous chlorine dioxide, high relative humidity, and mild heat.

PubMed

Bang, Jihyun; Choi, Moonhak; Son, Hyeri; Beuchat, Larry R; Kim, Yoonsook; Kim, Hoikyung; Ryu, Jee-Hoon

2016-11-21

Sanitizing radish seeds intended for edible sprout production was achieved by applying simultaneous treatments with gaseous chlorine dioxide (ClO 2 ), high relative humidity (RH, 100%), and mild heat (55°C). Gaseous ClO 2 was produced from aqueous ClO 2 (0.66ml) by mixing sulfuric acid (5% w/v) with sodium chlorite (10 mg/mL) in a sealed container (1.8L). Greater amounts of gaseous ClO 2 were measured at 23% RH (144ppm after 6h) than at 100% RH (66ppm after 6h); however, the lethal activity of gaseous ClO 2 against naturally occurring mesophilic aerobic bacteria (MAB) on radish seeds was significantly enhanced at 100% RH. For example, when exposed to gaseous ClO 2 at 23% RH, the number of MAB on radish seeds decreased from 3.7logCFU/g to 2.6logCFU/g after 6h. However, when exposed to gaseous ClO 2 at 100% RH for 6h, the MAB population decreased to 0.7logCFU/g after 6h. Gaseous ClO 2 was produced in higher amounts at 55°C than at 25°C, but decreased more rapidly over time at 55°C than at 25°C. The lethal activity of gaseous ClO 2 against MAB on radish seeds was greater at 55°C than at 25°C. When radish seeds were treated with gaseous ClO 2 (peak concentration: 195ppm) at 100% RH and 55°C, MAB were reduced to populations below the detectable level (<-0.7logCFU/g) within 2h without decreasing the seed germination rate (97.7%). The lethality of combined treatments against artificially inoculated Escherichia coli O157:H7 was also evaluated. When exposed to gaseous ClO 2 at 100% RH and 55°C for 6h, the initial number of E. coli O157:H7 (3.5logCFU/g) on radish seeds decreased to below the detection limit (0.7logCFU/g) by direct plating but it was not eliminated from seeds. The germination rate of radish seeds was not significantly (P>0.05) decreased after treatment for 6h. The information reported here will be useful when developing decontamination strategies for producing microbiologically safe radish seed sprouts. Copyright © 2016. Published by Elsevier B.V.

Deregulation of the arginine deiminase (arc) operon in penicillin-tolerant mutants of Streptococcus gordonii.

PubMed

Caldelari, I; Loeliger, B; Langen, H; Glauser, M P; Moreillon, P

2000-10-01

Penicillin tolerance is an incompletely understood phenomenon that allows bacteria to resist drug-induced killing. Tolerance was studied with independent Streptococcus gordonii mutants generated by cyclic exposure to 500 times the MIC of penicillin. Parent cultures lost 4 to 5 log(10) CFU/ml of viable counts/24 h. In contrast, each of four independent mutant cultures lost < or =2 log(10) CFU/ml/24 h. The mutants had unchanged penicillin-binding proteins but contained increased amounts of two proteins with respective masses of ca. 50 and 45 kDa. One mutant (Tol1) was further characterized. The two proteins showing increased levels were homologous to the arginine deiminase and ornithine carbamoyl transferase of other gram-positive bacteria and were encoded by an operon that was >80% similar to the arginine-deiminase (arc) operon of these organisms. Partial nucleotide sequencing and insertion inactivation of the S. gordonii arc locus indicated that tolerance was not a direct consequence of arc alteration. On the other hand, genetic transformation of tolerance by Tol1 DNA always conferred arc deregulation. In nontolerant recipients, arc was repressed during exponential growth and up-regulated during postexponential growth. In tolerant transformants, arc was constitutively expressed. Tol1 DNA transformed tolerance at the same rate as transformation of a point mutation (10(-2) to 10(-3)). The tolerance mutation mapped on a specific chromosomal fragment but was physically distant from arc. Importantly, arc deregulation was observed in most (6 of 10) of additional independent penicillin-tolerant mutants. Thus, although not exclusive, the association between arc deregulation and tolerance was not fortuitous. Since penicillin selection mimicked the antibiotic pressure operating in the clinical environment, arc deregulation might be an important correlate of naturally occurring tolerance and help in understanding the mechanism(s) underlying this clinically problematic phenotype.

Interactions of meat-associated bacteriocin-producing Lactobacilli with Listeria innocua under stringent sausage fermentation conditions.

PubMed

Leroy, Frédéric; Lievens, Kristoff; De Vuyst, Luc

2005-10-01

The kinetics of the antilisterial effect of meat-associated lactobacilli on Listeria innocua LMG 13568 were investigated during laboratory batch fermentations. During these fermentations, which were performed in a liquid meat simulation medium, a combination of process factors typical for European-style sausage fermentations was applied, such as a temperature of 20 degrees C and a representative pH and salting profile. Two bacteriocin-producing sausage isolates (Lactobacillus sakei CTC 494 and Lactobacillus curvatus LTH 1174), which have already proven efficacy in sausage trials, and one nonbacteriocinogenic, industrial strain (Lactobacillus sakei I), were evaluated. Staphylococcus carnosus 833 was included in the experiment because of its role in flavor and color development. When grown as a monoculture or upon cocultivation with L. sakei I and S. carnosus 833, L. innocua LMG 13568 developed slightly, despite the stress of low temperature, pH, lactic acid, salt, and nitrite. In contrast, when either of the bacteriocin producers was used, the L. innocua LMG 13568 population was rapidly inactivated with more than 3 log CFU ml(-1) after 2 days of fermentation. A bacteriocin-tolerant L. innocua LMG 13568 subpopulation (4 X 10(-4)) remained after bacteriocin inactivation. Thus, when the initial level of L. innocua LMG 13568 equaled 3 log CFU ml(-1), all cells were inactivated and no bacteriocin-tolerant cells were detected, even after 7 days of incubation. S. carnosus was not inactivated by the Lactobacillus bacteriocins and displayed slight growth.

Optimization of antibacterial activity by Gold-Thread (Coptidis Rhizoma Franch) against Streptococcus mutans using evolutionary operation-factorial design technique.

PubMed

Choi, Ung-Kyu; Kim, Mi-Hyang; Lee, Nan-Hee

2007-11-01

This study was conducted to find the optimum extraction condition of Gold-Thread for antibacterial activity against Streptococcus mutans using The evolutionary operation-factorial design technique. Higher antibacterial activity was achieved in a higher extraction temperature (R2 = -0.79) and in a longer extraction time (R2 = -0.71). Antibacterial activity was not affected by differentiation of the ethanol concentration in the extraction solvent (R2 = -0.12). The maximum antibacterial activity of clove against S. mutans determined by the EVOP-factorial technique was obtained at 80 degrees C extraction temperature, 26 h extraction time, and 50% ethanol concentration. The population of S. mutans decreased from 6.110 logCFU/ml in the initial set to 4.125 logCFU/ml in the third set.

Lethality of home-style dehydrator processes against Escherichia coli O157:H7 and salmonella serovars in the manufacture of ground-and-formed beef jerky and the potential for using a pathogen surrogate in process validation.

PubMed

Borowski, A G; Ingham, S C; Ingham, B H

2009-10-01

Ground-and-formed beef jerky can be made easily at home with ground beef and kits that include spice, cure, and jerky-forming equipment. Ground beef poses inherent risks of illness due to Escherichia coli O157:H7 and Salmonella contamination, making adequate pathogen lethality important in jerky manufacturing. We evaluated the effectiveness of drying regimes at eliminating E. coli O157:H7 and Salmonella in seasoned ground-and-formed beef jerky manufactured with three home-style dehydrators and one small commercial unit. Inoculated jerky strips were dried for up to 12 or 24 h in a home-style or the commercial unit, respectively, with target drying temperatures ranging from 51.7 degrees C (125 degrees F) to 71.1 degrees C (160 degrees F). Pathogen lethality varied with seasoning, temperature, and drying time (n = 288 samples). Lethality against E. coli O157:H7 ranged from 1.5 log CFU (Jerky Xpress, 57.2 degrees C [135 degrees F], 4 h) to 6.4 log CFU (Gardenmaster, 68.3 degrees C [155 degrees F], 12 h), and varied with seasoning. Lethality against Salmonella ranged from 1.7 log CFU (Jerky Xpress, 57.2 degrees C [135 degrees F], 4 h) to 6.0 log CFU (Gardenmaster, 68.3 degrees C [155 degrees F], 12 h), and also varied with seasoning. There was a > or =5-log CFU reduction in both pathogens in 0, 10, and 27 % of samples at 4, 8, and 12 h, respectively. Heating jerky for 10 min at 135 degrees C (275 degrees F) 4 or 6 h postdrying increased lethality, on average, 2.99 log CFU for Salmonella and 3.02 log CFU for E. coli O157:H7. The use of a lactic acid bacterium culture (Pediococcus spp.) as a pathogen surrogate accurately predicted safety in 28 % of samples containing E. coli O157:H7 and 78% of Salmonella-inoculated samples.

Antibacterial Effects of Levofloxacin, Erythromycin, and Rifampin in a Human Monocyte System against Legionella pneumophila

PubMed Central

Baltch, Aldona L.; Smith, Raymond P.; Franke, Mary A.; Michelsen, Phyllis B.

1998-01-01

The antibacterial activities of levofloxacin, erythromycin, and rifampin against intracellular Legionella pneumophila L-1033, serogroup 1, were studied. In an in vitro system utilizing adherent human monocytes, L. pneumophila L-1033, a phagocytosis time period of 1 h, and antibiotic (levofloxacin, erythromycin, and/or rifampin) at 1 to 10 times the MIC, the CFU/ml values for the monocyte lysate were determined during 0- to 4-day time periods. The decrease in CFU/ml with levofloxacin at pH 7.4 was rapid, occurring within 24 h, and was drug concentration dependent (P < 0.01). The decrease in CFU with rifampin was first observed at 48 h (P < 0.01), while only a minimal decrease in CFU/ml was observed with erythromycin. Combination of levofloxacin and rifampin and of levofloxacin and erythromycin at ten times their MICs significantly decreased the CFU/ml value (P < 0.01), to the value attained by levofloxacin alone, while combination of rifampin and erythromycin did not. Removal of levofloxacin after 24 h of incubation resulted in regrowth of L. pneumophila L-1033, while a continued slow decrease in CFU/ml was seen following rifampin removal; CFU/ml values were unaffected by the removal of erythromycin. At 4 days, and even in assays performed following antibiotic removal, the CFU/ml value continued to be lower in the levofloxacin and rifampin assays than in the assays with erythromycin. Levofloxacin had a significantly higher bactericidal activity against L. pneumophila L-1033 than erythromycin or rifampin. In these assays, the addition of erythromycin or rifampin did not affect the antibacterial activity of levofloxacin. PMID:9835507

Prevalence of Vibrio vulnificus and Vibrio parahaemolyticus in the Maryland Coastal Bays

NASA Astrophysics Data System (ADS)

De Pascuale, V. O.

2016-02-01

The bacterial family of Vibrionaceae is indigenous in the marine estuarine environments such as the Maryland Coastal Bays. Vibrio vulnificus and Vibrio parahaemolyticus are both pathogenic bacteria. Understanding the distribution of Vibrio species is crucial because of the health concerns associated with the bacteria. The aim of this study was to evaluate the overall abundance of bacteria with a focus on Vibrio species in the Maryland Coastal Bays. Seawater samples were collected from 10 different sites that differ with regard to water quality. The total bacteria count (TBC) was determined by two methods: Total plate count and Epifluorescence microscopy. The most-probable-number (MPN) methodology was used to estimate the population of Vibrio parahaemolyticus and Vibrio vulnificus. In addition to the bacteriological analysis, the environmental parameters of temperature and salinity were measured using YSI 6600 multiparameter meter. The average total bacteria count was 2.21 log CFU ml-1. Vibrio vulnificus comprised 5% of the total bacteria count while Vibrio parahaemolyticus comprised only 2% of the total bacteria count. Vibrio vulnificus ranged from 0.30 to 2.48 log MPN ml-1 at the sites tested. Lower Vibrio parahaemolyticus count was observed at the sites with a range of 0.30 to 1.97 log MPN ml-1. There was no significant correlation between the environmental parameters and the Vibrio spp. Since both Vibrio vulnificus and Vibrio parahaemolyticus peak in the summer, there is a potential for a risk of wound infections and gastrointestinal illness based on this data.

Microbial fouling community analysis of the cooling water system of a nuclear test reactor with emphasis on sulphate reducing bacteria.

PubMed

Balamurugan, P; Joshi, M Hiren; Rao, T S

2011-10-01

Culture and molecular-based techniques were used to characterize bacterial diversity in the cooling water system of a fast breeder test reactor (FBTR). Techniques were selected for special emphasis on sulphate-reducing bacteria (SRB). Water samples from different locations of the FBTR cooling water system, in addition to biofilm scrapings from carbon steel coupons and a control SRB sample were characterized. Whole genome extraction of the water samples and SRB diversity by group specific primers were analysed using nested PCR and denaturing gradient gel electrophoresis (DGGE). The results of the bacterial assay in the cooling water showed that the total culturable bacteria (TCB) ranged from 10(3) to 10(5) cfu ml(-1); iron-reducing bacteria, 10(3) to 10(5) cfu ml(-1); iron oxidizing bacteria, 10(2) to 10(3) cfu ml(-1) and SRB, 2-29 cfu ml(-1). However, the counts of the various bacterial types in the biofilm sample were 2-3 orders of magnitude higher. SRB diversity by the nested PCR-DGGE approach showed the presence of groups 1, 5 and 6 in the FBTR cooling water system; however, groups 2, 3 and 4 were not detected. The study demonstrated that the PCR protocol influenced the results of the diversity analysis. The paper further discusses the microbiota of the cooling water system and its relevance in biofouling.

Development of cereal-based functional food using cereal-mix substrate fermented with probiotic strain - Pichia kudriavzevii OG32.

PubMed

Ogunremi, Omotade R; Agrawal, Renu; Sanni, Abiodun I

2015-11-01

Probiotic strains contribute to the functionality of foods during fermentation. In this present work, cereal-mix was fermented with probiotic Pichia kudriavzevii OG32. Selected fermentation parameters and functional properties of the product were determined. The growth of Pichia kudriavzevii OG32 was supported by the cereal-mix containing 1% salt and 0.2% red chili powder to counts of between 7.46 and 8.22 Log10 cfu/mL within 24 h. Pichia kudriavzevii OG32 increased the viscosity of cereal-mix with the highest inoculum size (1.84x105cfu/ml) giving the highest viscosity of 1793.6 mPa.S. An inoculum size of 1.98 × 10(4) cfu/mL gave the most acceptable product based on the sensory evaluation by the panelist. Forty volatile compounds were identified in the fermented product, while acids (32.21%) and esters (32.37%) accounted for the largest proportions. The cereal-based fermented product scavenged DPPH from 200 μmol/L methanolic solution by 55.71%. Probiotic yeast improved the sensory and some functional properties of cereal-based substrate during fermentation. This is one of the first reports on the volatile composition of cereal-based functional food produced with probiotic yeast.

Antimicrobial effect of electrolyzed oxidizing water against Escherichia coli O157:H7 and Listeria monocytogenes on fresh strawberries (Fragaria x ananassa).

PubMed

Udompijitkul, P; Daeschel, M A; Zhao, Y

2007-11-01

Antibacterial activity of electrolyzed oxidizing (EO) water prepared from 0.05% or 0.10% (w/v) sodium chloride (NaCl) solutions against indigenous bacteria associated with fresh strawberries (Fragaria x ananassa) was evaluated. The efficacy of EO water and sodium hypochlorite (NaOCl) solution in eliminating and controlling the growth of Listeria monocytogenes and Escherichia coli O157:H7 inoculated onto strawberries stored at 4 +/- 1 degrees C up to 15 d was investigated at exposure time of 1, 5, or 10 min. Posttreatment neutralization of fruit surfaces was also determined. More than 2 log(10) CFU/g reductions of aerobic mesophiles were obtained in fruits washed for 10 or 15 min in EO water prepared from 0.10% (w/v) NaCl solution. Bactericidal activity of the disinfectants against L. monocytogenes and E. coli O157:H7 was not affected by posttreatment neutralization, and increasing exposure time did not significantly increase the antibacterial efficacy against both pathogens. While washing fruit surfaces with distilled water resulted in 1.90 and 1.27 log(10) CFU/mL of rinse fluid reduction of L. monocytogenes and E. coli O157:H7, respectively, >/= 2.60 log(10) CFU/mL of rinse fluid reduction of L. monocytogenes and up to 2.35 and 3.12 log(10) CFU/mL of rinse fluid reduction of E. coli O157:H7 were observed on fruit surfaces washed with EO water and NaOCl solution, respectively. Listeria monocytogenes and E. coli O157:H7 populations decreased over storage regardless of prior treatment. However, EO water and aqueous NaOCl did not show higher antimicrobial potential than water treatment during refrigeration storage.

Bacterial Growth Kinetics under a Novel Flexible Methacrylate Dressing Serving as a Drug Delivery Vehicle for Antiseptics

PubMed Central

Forstner, Christina; Leitgeb, Johannes; Schuster, Rupert; Dosch, Verena; Kramer, Axel; Cutting, Keith F.; Leaper, David J.; Assadian, Ojan

2013-01-01

A flexible methacrylate powder dressing (Altrazeal®) transforms into a wound contour conforming matrix once in contact with wound exudate. We hypothesised that it may also serve as a drug delivery vehicle for antiseptics. The antimicrobial efficacy and influence on bacterial growth kinetics in combination with three antiseptics was investigated in an in vitro porcine wound model. Standardized in vitro wounds were contaminated with Staphylococcus aureus (MRSA; ATCC 33591) and divided into six groups: no dressing (negative control), methacrylate dressing alone, and combinations with application of 0.02% Polyhexamethylene Biguanide (PHMB), 0.4% PHMB, 0.1% PHMB + 0.1% betaine, 7.7 mg/mL Povidone-iodine (PVP-iodine), and 0.1% Octenidine-dihydrochloride (OCT) + 2% phenoxyethanol. Bacterial load per gram tissue was measured over five days. The highest reduction was observed with PVP-iodine at 24 h to log10 1.43 cfu/g, followed by OCT at 48 h to log10 2.41 cfu/g. Whilst 0.02% PHMB resulted in a stable bacterial load over 120 h to log10 4.00 cfu/g over 120 h, 0.1% PHMB + 0.1% betaine inhibited growth during the first 48 h, with slightly increasing bacterial numbers up to log10 5.38 cfu/g at 120 h. These results indicate that this flexible methacrylate dressing can be loaded with various antiseptics serving as drug delivery system. Depending on the selected combination, an individually shaped and controlled antibacterial effect may be achieved using the same type of wound dressing. PMID:23698780

Inhibition of Clostridium perfringens spore germination and outgrowth by lemon juice and vinegar product in reduced NaCl roast beef.

PubMed

Li, Lin; Valenzuela-Martinez, Carol; Redondo, Mauricio; Juneja, Vijay K; Burson, Dennis E; Thippareddi, Harshavardhan

2012-11-01

Inhibition of Clostridium perfringens spore germination and outgrowth in reduced sodium roast beef by a blend of buffered lemon juice concentrate and vinegar (MoStatin LV1) during abusive exponential cooling was evaluated. Roast beef containing salt (NaCl; 1%, 1.5%, or 2%, w/w), blend of sodium pyro- and poly-phosphates (0.3%), and MoStatin LV1 (0%, 2%, or 2.5%) was inoculated with a 3-strain C. perfringens spore cocktail to achieve final spore population of 2.5 to 3.0 log CFU/g. The inoculated products were heat treated and cooled exponentially from 54.4 to 4.4 °C within 6.5, 9, 12, 15, 18, or 21 h. Cooling of roast beef (2.0% NaCl) within 6.5 and 9 h resulted in <1.0 log CFU/g increase in C. perfringens spore germination and outgrowth, whereas reducing the salt concentration to 1.5% and 1.0% resulted in >1.0 log CFU/g increase for cooling times longer than 9 h (1.1 and 2.2 log CFU/g, respectively). Incorporation of MoStatin LV1 into the roast beef formulation minimized the C. perfringens spore germination and outgrowth to <1.0 log CFU/g, regardless of the salt concentration and the cooling time. Cooked, ready-to-eat meat products should be cooled rapidly to reduce the risk of Clostridium perfringens spore germination and outgrowth. Meat processors are reducing the sodium chloride content of the processed meats as a consequence of the dietary recommendations. Sodium chloride reduces the risk of C. perfringens spore germination and outgrowth in meat products. Antimicrobials that contribute minimally to the sodium content of the product should be incorporated into processed meats to assure food safety. Buffered lemon juice and vinegar can be incorporated into meat product formulations to reduce the risk of C. perfringens spore germination and outgrowth during abusive cooling. Journal of Food Science © 2012 Institute of Food Technologists® No claim to original US government works.

Persistence of Mycobacterium paratuberculosis during Manufacture and Ripening of Cheddar Cheese

PubMed Central

Donaghy, J. A.; Totton, N. L.; Rowe, M. T.

2004-01-01

Model Cheddar cheeses were prepared from pasteurized milk artificially contaminated with high 104 to 105 CFU/ml) and low (101 to 102 CFU/ml) inocula of three different Mycobacterium paratuberculosis strains. A reference strain, NCTC 8578, and two strains (806PSS and 796PSS) previously isolated from pasteurized milk for retail sale were investigated in this study. The manufactured Cheddar cheeses were similar in pH, salt, moisture, and fat composition to commercial Cheddar. The survival of M. paratuberculosis cells was monitored over a 27-week ripening period by plating homogenized cheese samples onto HEYM agar medium supplemented with the antibiotics vancomycin, amphotericin B, and nalidixic acid without a decontamination step. A concentration effect was observed in M. paratuberculosis numbers between the inoculated milk and the 1-day old cheeses for each strain. For all manufactured cheeses, a slow gradual decrease in M. paratuberculosis CFU in cheese was observed over the ripening period. In all cases where high levels (>3.6 log10) of M. paratuberculosis were present in 1-day cheeses, the organism was culturable after the 27-week ripening period. The D values calculated for strains 806PSS, 796PSS, and NCTC 8578 were 107, 96, and 90 days, respectively. At low levels of contamination, M. paratuberculosis was only culturable from 27-week-old cheese spiked with strain 806PSS. M. paratuberculosis was recovered from the whey fraction in 10 of the 12 manufactured cheeses. Up to 4% of the initial M. paratuberculosis load was recovered in the culture-positive whey fractions at either the high or low initial inoculum. PMID:15294829

Growth and Survival of Acid-Resistant and Non-Acid-Resistant Shiga-Toxin-Producing Escherichia coli Strains during the Manufacture and Ripening of Camembert Cheese.

PubMed

Montet, M P; Jamet, E; Ganet, S; Dizin, M; Miszczycha, S; Dunière, L; Thevenot, D; Vernozy-Rozand, C

2009-01-01

Growth and survival of acid-resistant (AR) and non-acid-resistant (NAR) Shiga-toxin-producing Escherichia coli (STEC) strains were investigated during the manufacture and ripening of microfiltered milk Camembert cheeses. The induction of acid resistance of the STEC strains in cheeses was also studied. Six different mixtures of AR and/or NAR STEC strains were inoculated separately into microfiltered milk at a level of 10(3) CFU mL(-1). The STEC counts (AR and NAR) initially increased by 1 to 2 log(10) CFU g(-1) during cheese-making. Thereafter, the populations stabilized during salting/drying and then decreased during the early stages of ripening. Exposing the STEC strains in artificially inoculated cheeses to simulated gastric fluid (SGF - pH: 2.0) reduced the number of NAR strains to undetectable levels within 40 minutes, versus 120 minutes for the AR STEC strains. AR and NAR STEC were able to survive during the manufacture and ripening of Camembert cheese prepared from microfiltered milk with no evidence of induced acid tolerance in NAR STEC strains.

Microorganisms as an Indicator of Hygiene Status Among Migrant Food Handlers in Peninsular Malaysia.

PubMed

Woh, Pei Yee; Thong, Kwai Lin; Lim, Yvonne Ai Lian; Behnke, Jerzy Marian; Lewis, John Watkin; Mohd Zain, Siti Nursheena

2017-10-01

This study used microbial indicators to assess the hygiene status of 383 migrant food handlers from 3 urban cities in Peninsular Malaysia. Microbiological analysis revealed that all the hand swabs tested 99.5% positive for aerobic plate counts (mean [M] ± standard deviation [SD] = 3.57 ± 0.83 log 10 CFU [colony forming unit]), 20.8% positive for total coliform/ Escherichia coli (M ± SD = 0.30 ± 0.67 log 10 CFU), and 63.4% positive for Staphylococcus aureus (M ± SD = 1.38 ± 1.26 log 10 CFU). In addition, aerobic plate counts and Staphylococcus aureus counts exceeded the acceptable standard levels. Bacterial counts were found to be significantly associated with subjects' country of origin ( P = .019) and working responsibilities ( P = .001). Our findings indicate high probability of transmission of pathogenic bacteria from the food handlers' hands to customers during meal preparation and serving. This calls for improvements in personal hygiene and sanitation standards by the relevant health authorities among migrant food handlers.

Microbiological hazard identification and exposure assessment of poultry products sold in various localities of Hyderabad, India.

PubMed

Sudershan, Rao V; Naveen Kumar, R; Kashinath, L; Bhaskar, V; Polasa, K

2012-01-01

A study was carried out to identify microbiological hazards and assess their exposure associated with consumption of poultry based street food served in different localities of Hyderabad. The study indicated that chicken 65, chicken fried rice, chicken noodles, chicken Manchuria and chilly chicken are the most common recipes. A process flow diagram was developed to identify critical control points in the food item. After analysis of the samples at each level of preparation, it was observed that rice and noodles were kept at room temperature for about 5-6 hrs which was a critical control point. A total of 376 samples including chicken fried rice, chicken noodles, boiled noodles and boiled rice were collected from circle 1, 2, 3, 4, 5, 6, and 7 of Greater Hyderabad municipal corporation (GHMC) and analyzed for microbiological examination. The most prevalent pathogenic bacteria isolated were S. aureus (3.4 log 10 cfu/g) and B. cereus (3.4 log 10 cfu/g). Salmonella spp. was present in salads (3.2 log 10 cfu/g) and hand washings of the food handler (3.5 log 10 cfu/g). Salmonella contamination was found in salads served along with chicken fried rice and chicken noodles than in the food.

The effect of ice-cream-scoop water on the hygiene of ice cream.

PubMed Central

Wilson, I. G.; Heaney, J. C.; Weatherup, S. T.

1997-01-01

A survey of unopened ice cream, ice cream in use, and ice-cream-scoop water (n = 91) was conducted to determine the effect of scoop water hygiene on the microbiological quality of ice cream. An aerobic plate count around 10(6) c.f.u. ml-1 was the modal value for scoop waters. Unopened ice creams generally had counts around 10(3)-10(4) c.f.u. ml-1 and this increased by one order of magnitude when in use. Many scoop waters had low coliform counts, but almost half contained > 100 c.f.u. ml-1. E. coli was isolated in 18% of ice creams in use, and in 10% of unopened ice creams. S. aureus was not detected in any sample. Statistical analysis showed strong associations between indicator organisms and increased counts in ice cream in use. EC guidelines for indicator organisms in ice cream were exceeded by up to 56% of samples. PMID:9287941

The effect of ice-cream-scoop water on the hygiene of ice cream.

PubMed

Wilson, I G; Heaney, J C; Weatherup, S T

1997-08-01

A survey of unopened ice cream, ice cream in use, and ice-cream-scoop water (n = 91) was conducted to determine the effect of scoop water hygiene on the microbiological quality of ice cream. An aerobic plate count around 10(6) c.f.u. ml-1 was the modal value for scoop waters. Unopened ice creams generally had counts around 10(3)-10(4) c.f.u. ml-1 and this increased by one order of magnitude when in use. Many scoop waters had low coliform counts, but almost half contained > 100 c.f.u. ml-1. E. coli was isolated in 18% of ice creams in use, and in 10% of unopened ice creams. S. aureus was not detected in any sample. Statistical analysis showed strong associations between indicator organisms and increased counts in ice cream in use. EC guidelines for indicator organisms in ice cream were exceeded by up to 56% of samples.

Biocontrol of Escherichia coli O157: H7 on fresh-cut leafy greens.

PubMed

Boyacioglu, Olcay; Sharma, Manan; Sulakvelidze, Alexander; Goktepe, Ipek

2013-01-01

The effect of a bacteriophage cocktail (EcoShield™) that is specific against Escherichia coli O157:H7 was evaluated against a nalidixic acid-resistant enterohemorrhagic E. coli O157:H7 RM4407 (EHEC) strain on leafy greens stored under either (1) ambient air or (2) modified atmosphere (MA; 5% O 2 /35% CO 2 /60% N 2 ). Pieces (~2 × 2 cm 2 ) of leafy greens (lettuce and spinach) inoculated with 4.5 log CFU/cm 2 EHEC were sprayed with EcoShield™ (6.5 log PFU/cm 2 ). Samples were stored at 4 or 10°C for up to 15 d. On spinach, the level of EHEC declined by 2.38 and 2.49 log CFU/cm 2 at 4 and 10°C, respectively, 30 min after phage application (p ≤ 0.05). EcoShield™ was also effective in reducing EHEC on the surface of green leaf lettuce stored at 4°C by 2.49 and 3.28 log units in 30 min and 2 h, respectively (p ≤ 0.05). At 4°C under atmospheric air, the phage cocktail significantly (p ≤ 0.05) lowered the EHEC counts in one day by 1.19, 3.21 and 3.25 log CFU/cm 2 on spinach, green leaf and romaine lettuce, respectively compared with control (no bacteriophage) treatments. When stored under MA at 4°C, phages reduced (p ≤ 0.05) EHEC populations by 2.18, 3.50 and 3.13 log CFU/cm 2 , on spinach, green leaf and romaine lettuce. At 10°C, EHEC reductions under atmospheric air storage were 1.99, 3.90 and 3.99 log CFU/cm 2 (p ≤ 0.05), while population reductions under MA were 3.08, 3.89 and 4.34 logs on spinach, green leaf and romaine lettuce, respectively, compared with controls (p ≤ 0.05). The results of this study showed that bacteriophages were effective in reducing the levels of E. coli O157:H7 on fresh leafy produce, and that the reduction was further improved when produce was stored under the MA conditions.

Inactivation of Escherichia coli O157:H7 in nonintact beefsteaks of different thicknesses cooked by pan broiling, double pan broiling, or roasting by using five types of cooking appliances.

PubMed

Shen, Cangliang; Adler, Jeremy M; Geornaras, Ifigenia; Belk, Keith E; Smith, Gary C; Sofos, John N

2010-03-01

This study compared thermal inactivation of Escherichia coli O157:H7 in nonintact beefsteaks of different thicknesses by different cooking methods and appliances. Coarsely ground beef was inoculated with rifampin-resistant E. coli O157:H7 (eight-strain composite, 6 to 7 log CFU/g) and then mixed with sodium chloride (0.45%) plus sodium tripolyphosphate (0.23%); the total water added was 10%. The meat was stuffed into bags (10-cm diameter), semifrozen (-20 degrees C, 6 h), and cut into 1.5-, 2.5-, and 4.0-cm-thick steaks. Samples were then individually vacuum packaged, frozen (-20 degrees C, 42 h), and tempered (4 degrees C, 2.5 h) before cooking. Partially thawed (-2 +/- 1 degrees C) steaks were pan broiled (Presto electric skillet and Sanyo grill), double pan broiled (George Foreman grill), or roasted (Oster toaster oven and Magic Chef standard kitchen oven) to a geometric center temperature of 65 degrees C. Extent of pathogen inactivation decreased in order of roasting (2.0 to 4.2 log CFU/g) > pan broiling (1.6 to 2.8 log CFU/g) >/= double pan broiling (1.1 to 2.3 log CFU/g). Cooking of 4.0-cm-thick steaks required a longer time (19.8 to 65.0 min; variation was due to different cooking appliances), and caused greater reductions in counts (2.3 to 4.2 log CFU/g) than it did in thinner samples (1.1 to 2.9 log CFU/g). The time to reach the target temperature increased in order of George Foreman grill (3.9 to 19.8 min) < Oster toaster oven (11.3 to 45.0 min) < Presto electric skillet (16.3 to 55.0 min) < Sanyo grill (14.3 to 65.0 min) < standard kitchen oven (20.0 to 63.0 min); variation was due to steak thickness. Results indicated that increased steak thickness allowed greater inactivation of E. coli O157:H7, as time to reach the target internal temperature increased. Roasting in a kitchen oven was most effective for pathogen inactivation.

Optimization of Synergistic Combination Regimens against Carbapenem- and Aminoglycoside-Resistant Clinical Pseudomonas aeruginosa Isolates via Mechanism-Based Pharmacokinetic/Pharmacodynamic Modeling

PubMed Central

Yadav, Rajbharan; Nation, Roger L.

2016-01-01

ABSTRACT Optimizing antibiotic combinations is promising to combat multidrug-resistant Pseudomonas aeruginosa. This study aimed to systematically evaluate synergistic bacterial killing and prevention of resistance by carbapenem and aminoglycoside combinations and to rationally optimize combination dosage regimens via a mechanism-based mathematical model (MBM). We studied monotherapies and combinations of imipenem with tobramycin or amikacin against three difficult-to-treat double-resistant clinical P. aeruginosa isolates. Viable-count profiles of total and resistant populations were quantified in 48-h static-concentration time-kill studies (inoculum, 107.5 CFU/ml). We rationally optimized combination dosage regimens via MBM and Monte Carlo simulations against isolate FADDI-PA088 (MIC of imipenem [MICimipenem] of 16 mg/liter and MICtobramycin of 32 mg/liter, i.e., both 98th percentiles according to the EUCAST database). Against this isolate, imipenem (1.5× MIC) combined with 1 to 2 mg/liter tobramycin (MIC, 32 mg/liter) or amikacin (MIC, 4 mg/liter) yielded ≥2-log10 more killing than the most active monotherapy at 48 h and prevented resistance. For all three strains, synergistic killing without resistance was achieved by ≥0.88× MICimipenem in combination with a median of 0.75× MICtobramycin (range, 0.032× to 2.0× MICtobramycin) or 0.50× MICamikacin (range, 0.25× to 0.50× MICamikacin). The MBM indicated that aminoglycosides significantly enhanced the imipenem target site concentration up to 3-fold; achieving 50% of this synergistic effect required aminoglycoside concentrations of 1.34 mg/liter (if the aminoglycoside MIC was 4 mg/liter) and 4.88 mg/liter (for MICs of 8 to 32 mg/liter). An optimized combination regimen (continuous infusion of imipenem at 5 g/day plus a 0.5-h infusion with 7 mg/kg of body weight tobramycin) was predicted to achieve >2.0-log10 killing and prevent regrowth at 48 h in 90.3% of patients (median bacterial killing, >4.0 log10 CFU/ml) against double-resistant isolate FADDI-PA088 and therefore was highly promising. PMID:27821448

Reduction of Salmonella enterica on alfalfa seeds with acidic electrolyzed oxidizing water and enhanced uptake of acidic electrolyzed oxidizing water into seeds by gas exchange.

PubMed

Stan, Silvia D; Daeschel, Mark A

2003-11-01

Alfalfa sprouts have been implicated in several salmonellosis outbreaks in recent years. The disinfectant effects of acidic electrolyzed oxidizing (EO) water against Salmonella enterica both in an aqueous system and on artificially contaminated alfalfa seeds were determined. The optimum ratio of seeds to EO water was determined in order to maximize the antimicrobial effect of EO water. Seeds were combined with EO water at ratios (wt/vol) of 1:4, 1:10, 1:20, 1:40, and 1:100, and the characteristics of EO water (pH, oxidation reduction potential [ORP], and free chlorine concentration) were determined. When the ratio of seeds to EO water was increased from 1:4 to 1:100, the pH decreased from 3.82 to 2.63, while the ORP increased from +455 to +1,073 mV. EO water (with a pH of 2.54 to 2.38 and an ORP of +1,083 to +1,092 mV) exhibited strong potential for the inactivation of S. enterica in an aqueous system (producing a reduction of at least 6.6 log CFU/ml). Treatment of artificially contaminated alfalfa seeds with EO water at a seed-to-EO water ratio of 1:100 for 15 and 60 min significantly reduced Salmonella populations by 2.04 and 1.96 log CFU/g, respectively (P < 0.05), while a Butterfield's buffer wash decreased Salmonella populations by 0.18 and 0.23 log CFU/g, respectively. After treatment, EO water was Salmonella negative by enrichment with or without neutralization. Germination of seeds was not significantly affected (P > 0.05) by treatment for up to 60 min in electrolyzed water. The uptake of liquid into the seeds was influenced by the internal gas composition (air, N2, or O2) of seeds before the liquid was added.

Effects of lactic acid and commercial chilling processes on survival of Salmonella, Yersinia enterocolitica, and Campylobacter coli in pork variety meats.

PubMed

King, Amanda M; Miller, Rhonda K; Castillo, Alejandro; Griffin, Davey B; Hardin, Margaret D

2012-09-01

Current industry chilling practices with and without the application of 2% L-lactic acid were compared for their effectiveness at reducing levels of Salmonella, Yersinia enterocolitica, and Campylobacter coli on pork variety meats. Pork variety meats (livers, intestines, hearts, and stomachs) were inoculated individually with one of the three pathogens and subjected to five different treatment combinations that included one or more of the following: water wash (25°C), lactic acid spray (2%, 40 to 50°C), chilling (4°C), and freezing (-15°C). Samples were analyzed before treatment, after each treatment step, and after 2, 4, and 6 months of frozen storage. Results showed that when a lactic acid spray was used in combination with water spray, immediate reductions were approximately 0.5 log CFU per sample of Salmonella, 0.8 log CFU per sample of Y. enterocolitica, and 1.1 log CFU per sample of C. coli. Chilling, both alone and in combination with spray treatments, had little effect on pathogens, while freezing resulted in additional 0.5-log CFU per sample reductions in levels of Salmonella and Y. enterocolitica, and an additional 1.0-log CFU per sample reduction in levels of C. coli. While reductions of at least 1 log CFU per sample were observed on variety meats treated with only a water wash and subsequently frozen, samples treated with lactic acid had greater additional reductions than those treated with only a water spray throughout frozen storage. The results of this study suggest that the use of lactic acid as a decontamination intervention, when used in combination with good manufacturing practices during processing, causes significant reductions in levels of Salmonella, Y. enterocolitica, and C. coli on pork variety meats.

Determination of foodborne pathogenic bacteria by multiplex PCR-microchip capillary electrophoresis with genetic algorithm-support vector regression optimization.

PubMed

Li, Yongxin; Li, Yuanqian; Zheng, Bo; Qu, Lingli; Li, Can

2009-06-08

A rapid and sensitive method based on microchip capillary electrophoresis with condition optimization of genetic algorithm-support vector regression (GA-SVR) was developed and applied to simultaneous analysis of multiplex PCR products of four foodborne pathogenic bacteria. Four pairs of oligonucleotide primers were designed to exclusively amplify the targeted gene of Vibrio parahemolyticus, Salmonella, Escherichia coli (E. coli) O157:H7, Shigella and the quadruplex PCR parameters were optimized. At the same time, GA-SVR was employed to optimize the separation conditions of DNA fragments in microchip capillary electrophoresis. The proposed method was applied to simultaneously detect the multiplex PCR products of four foodborne pathogenic bacteria under the optimal conditions within 8 min. The levels of detection were as low as 1.2 x 10(2) CFU mL(-1) of Vibrio parahemolyticus, 2.9 x 10(2) CFU mL(-1) of Salmonella, 8.7 x 10(1) CFU mL(-1) of E. coli O157:H7 and 5.2 x 10(1) CFU mL(-1) of Shigella, respectively. The relative standard deviation of migration time was in the range of 0.74-2.09%. The results demonstrated that the good resolution and less analytical time were achieved due to the application of the multivariate strategy. This study offers an efficient alternative to routine foodborne pathogenic bacteria detection in a fast, reliable, and sensitive way.

Antibacterial activity of cinnamaldehyde and Sporan against Escherichia coli O157:H7 and Salmonella

USDA-ARS?s Scientific Manuscript database

The in vitro antimicrobial effect of cinnamaldehyde and Sporan in combination with acetic acid against E. coli O157:H7 and Salmonella was investigated. A five strain cocktail of E. coli O157:H7 and Salmonella were inoculated in Luria-Bertoni broth (LB broth, 7 log CFU ml-1) containing cinnamaldehyde...

Hand contamination during routine care in medical wards: the role of hand hygiene compliance.

PubMed

Monistrol, Olga; López, M Liboria; Riera, Montserrat; Font, Roser; Nicolás, Carme; Escobar, Miguel Angel; Freixas, Núria; Garau, Javier; Calbo, Esther

2013-04-01

The hands of healthcare workers (HCWs) are the most common vehicle for the transmission of micro-organisms from patient to patient and within the healthcare environment. The aim of this study was to evaluate the impact of a multimodal campaign on the type and amount of resident and transient flora and the presence of potential risk factors for hand contamination during routine care. A before-after (PRE and POST periods) interventional study was carried out in medical wards of a tertiary care hospital. Eighty-nine samples were analysed. Samples were cultured immediately before patient contact using a glove-juice method. Data collected included socio-demographic and risk factors for hand contamination. Flora was measured as log10 c.f.u. ml(-1) and evaluated by comparing median values in the PRE and POST periods. Transient flora was isolated from the hands of 67.4 and 46.1 % of HCWs in the PRE and POST periods, respectively (P<0.001). Enterobacteriaceae, Pseudomonas spp. and meticillin-sensitive Staphylococcus aureus were the predominant contaminants. Resident flora was isolated from 92.1 % of HCWs in the PRE period and from 70.8 % in the POST period (P<0.001). The meticillin-resistant coagulase-negative staphylococci log10 c.f.u. count ml(-1) decreased from 1.96 ± 1.2 to 0.89 ± 1.2 (mean ± s d; P<0.001), and the global flora count decreased from 2.77 ± 1.1 to 1.56 ± 1.4 (P<0.001). In the POST period, the wearing of fewer rings (P<0.001), shorter fingernail length (P = 0.008), a shorter time since recent hand hygiene (HH) (P = 0.007) and an increased use of alcohol-based hand rub instead of soap (P<0.001) were documented. The HH multimodal strategy reduced the number of risk factors and the level of HCW hand contamination.

Bacteriological quality of weaning food and drinking water given to children of market women in Nigeria: implications for control of diarrhoea.

PubMed

Iroegbu, C U; Ene-Obong, H N; Uwaegbute, A C; Amazigo, U V

2000-12-01

Bacteriological quality of weaning food and drinking water given to 2 groups of children aged < or = years was evaluated by estimating bacterial cell count. One group consisted of those taken to market and the other of those left at home in the care of older siblings or house-helps. Bacterial counts (geometric mean) ranged from 5.02 +/- 1.82 to 8.70 +/- 1.0 log10 cfu per g or mL of food, and from 1.15 +/- 1.67 to 6.53 +/- 0.81 log10 cfu per g or 100 mL of water. Analysis of variance showed no significant difference in counts between types of food and between meals (breakfast and lunch). Bacterial contamination increased significantly with storage time, and was, in all circumstances except the water samples, significantly higher in foods given to children left at home. Reheated leftover foods also had significantly higher bacterial load than the freshly-cooked food. Coliform count varied significantly with source of drinking water. Poor hygiene standard (inferred from bacterial contamination) was generally observed among mothers weaning < or = 2-year-old children, while they were engaged in trading activities in the market, thus exposing their children to high risk of diarrhoea. Hygiene was significantly poorer in weaning of children left at home in the care of older siblings or house-helps. This implies that, in spite of their trading activities in the market, mothers still take better care of their babies than the older siblings or house-helps who may be inexperienced. These mothers may need education on childcare and food hygiene to suit to their trading activities, for example, during their monthly meetings. There is also a need to establish ORT (oral rehydration therapy) corners in the markets as part of the municipal services. This can be used not only for efficient and quick management of diarrhoea in the market but also for reinforcing hygiene education.

Microbiological characteristics of "androlla", a Spanish traditional pork sausage.

PubMed

García Fontán, María C; Lorenzo, José M; Parada, Ana; Franco, Inmaculada; Carballo, Javier

2007-02-01

Counts of total aerobic mesophilic microflora, lactic acid bacteria, salt-tolerant microflora, Enterobacteriaceae, enterococci, moulds and yeasts, and staphylococci, and some physico-chemical parameters (total solids, NaCl and nitrate contents and pH and aw values) were determined in 20 units of "androlla", a traditional dry-fermented sausage made in the NW of Spain. In general, high counts of all the investigated microbial groups were observed, with average values of 8.99 +/- 0.46 log cfu/g for the total aerobic mesophilic microflora, 9.11 +/- 0.16 log cfu/g for the lactic acid bacteria, 6.87 +/- 0.68 log cfu/g for the salt-tolerant microflora, 2.80+/-1.85 log cfu/g for the Enterobacteriaceae, 3.25 +/- 1.86 log cfu/g for the enterococci, 4.30 +/- 1.73 log cfu/g for the moulds and yeasts, and 3.62 +/- 0.60 log cfu/g for the staphylococci. From MRS agar, SPC agar + 7.5% NaCl, VRBG agar, and KAA agar, 10 colonies were randomly taken from each androlla unit and from each culture medium. A total of 200 strains per culture medium were then identified using the classical methods. Among the isolates from MRS agar, Lactobacillus sakei predominated, followed by Lactobacillus curvatus, Lactobacillus alimentarius and Lactobacillus plantarum. Of the 200 isolates obtained from SPC agar + 7.5% NaCl, only 56 strains belonged to the Staphylococcaceae or Micrococcaceae families. Among the Staphylococcaceae, Staphylococcus xylosus was the main species, followed by Staph. epidermidis; Staph. equorum, Staph. capitis and Staph. saprophyticus were isolated in very low proportions. Among the Micrococcaceae, Micrococcus luteus predominated, followed by Micrococcus lylae, Kocuria varians and Kocuria kristinae. Of the 150 isolates obtained from VRBG agar, Hafnia alvei was the main species, followed by Serratia liquefaciens and Enterobacter amnigenus; six isolates were identified as Salmonella. Among the 190 isolates obtained from KAA agar, 122 were considered enterococci; 20 isolates were identified as Enterococcus faecium, one as Enterococcus faecalis and 101 as Enterococcus inter faecalis-faecium.

Combinatorial Pharmacodynamics of Ceftolozane-Tazobactam against Genotypically Defined β-Lactamase-Producing Escherichia coli: Insights into the Pharmacokinetics/Pharmacodynamics of β-Lactam–β-Lactamase Inhibitor Combinations

PubMed Central

Soon, Rachel L.; Lenhard, Justin R.; Bulman, Zackery P.; Holden, Patricia N.; Kelchlin, Pamela; Steenbergen, Judith N.; Friedrich, Lawrence V.; Forrest, Alan

2016-01-01

Despite a dearth of new agents currently being developed to combat multidrug-resistant Gram-negative pathogens, the combination of ceftolozane and tazobactam was recently approved by the Food and Drug Administration to treat complicated intra-abdominal and urinary tract infections. To characterize the activity of the combination product, time-kill studies were conducted against 4 strains of Escherichia coli that differed in the type of β-lactamase they expressed. The four investigational strains included 2805 (no β-lactamase), 2890 (AmpC β-lactamase), 2842 (CMY-10 β-lactamase), and 2807 (CTX-M-15 β-lactamase), with MICs to ceftolozane of 0.25, 4, 8, and >128 mg/liter with no tazobactam, and MICs of 0.25, 1, 4, and 8 mg/liter with 4 mg/liter tazobactam, respectively. All four strains were exposed to a 6 by 5 array of ceftolozane (0, 1, 4, 16, 64, and 256 mg/liter) and tazobactam (0, 1, 4, 16, and 64 mg/liter) over 48 h using starting inocula of 106 and 108 CFU/ml. While ceftolozane-tazobactam achieved bactericidal activity against all 4 strains, the concentrations of ceftolozane and tazobactam required for a ≥3-log reduction varied between the two starting inocula and the 4 strains. At both inocula, the Hill plots (R2 > 0.882) of ceftolozane revealed significantly higher 50% effective concentrations (EC50s) at tazobactam concentrations of ≤4 mg/liter than those at concentrations of ≥16 mg/liter (P < 0.01). Moreover, the EC50s at 108 CFU/ml were 2.81 to 66.5 times greater than the EC50s at 106 CFU/ml (median, 10.7-fold increase; P = 0.002). These promising results indicate that ceftolozane-tazobactam achieves bactericidal activity against a wide range of β-lactamase-producing E. coli strains. PMID:26729494

Photodynamic topical antimicrobial therapy for infected foot ulcers in patients with diabetes: a randomized, double-blind, placebo-controlled study--the D.A.N.T.E (Diabetic ulcer Antimicrobial New Topical treatment Evaluation) study.

PubMed

Mannucci, Edoardo; Genovese, Stefano; Monami, Matteo; Navalesi, Giovanni; Dotta, Francesco; Anichini, Roberto; Romagnoli, Fabio; Gensini, Gianfranco

2014-01-01

This study was designed to assess the antimicrobial effect and tolerability of a single dose of a photo-activated gel containing RLP068 in the treatment for infected foot ulcers in subjects with diabetes. A randomized, double-blind, parallel series, placebo-controlled phase IIa trial was performed with three concentrations of RLP068 (0.10, 0.30, and 0.50 %), measuring total and pathogen microbial load on Day 1 (before and 1 h after topical gel application and photoactivation with 689 nm red light), on Days 3, 8, and 15, as add-on to systemic treatment with amoxicillin and clavulanic acid. Blood samples were also drawn 1, 2, and 48 h after administration for the assessment of systemic drug absorption. The trial was performed on 62 patients aged ≥18 years, with type 1 or type 2 diabetes and infected foot ulcer, with an area of 2-15 cm(2) and a maximum diameter ≤4.6 cm. A dose-dependent reduction in total microbial load was observed (-1.92 ± 1.21, -2.94 ± 1.60, and -3.00 ± 1.82 LogCFU/ml for 0.10, 0.30, and 0.50 % RPL068 vs. -1.00 ± 1.02 LogCFU/ml with placebo) immediately after illumination, with a progressive fading of the effect during follow-up. No safety issues emerged from the analysis of adverse events. Systemic absorption of RLP068 was negligible. Photodynamic antimicrobial treatment with RLP068 of infected diabetic foot ulcers is well tolerated and produces a significant reduction in germ load. Further clinical trials are needed to verify the efficacy of this approach as add-on to systemic antibiotic treatment.

Antimicrobial activity of a novel adhesive containing chlorhexidine gluconate (CHG) against the resident microflora in human volunteers

PubMed Central

Carty, Neal; Wibaux, Anne; Ward, Colleen; Paulson, Daryl S.; Johnson, Peter

2014-01-01

Objectives To evaluate the antimicrobial activity of a new, transparent composite film dressing, whose adhesive contains chlorhexidine gluconate (CHG), against the native microflora present on human skin. Methods CHG-containing adhesive film dressings and non-antimicrobial control film dressings were applied to the skin on the backs of healthy human volunteers without antiseptic preparation. Dressings were removed 1, 4 or 7 days after application. The bacterial populations underneath were measured by quantitative cultures (cylinder-scrub technique) and compared with one another as a function of time. Results The mean baseline microflora recovery was 3.24 log10 cfu/cm2. The mean log reductions from baseline measured from underneath the CHG-containing dressings were 0.87, 0.78 and 1.30 log10 cfu/cm2 on days 1, 4 and 7, respectively, compared with log reductions of 0.67, −0.87 and −1.29 log10 cfu/cm2 from underneath the control film dressings. There was no significant difference between the log reductions of the two treatments on day 1, but on days 4 and 7 the log reduction associated with the CHG adhesive was significantly higher than that associated with the control adhesive. Conclusions The adhesive containing CHG was associated with a sustained antimicrobial effect that was not present in the control. Incorporating the antimicrobial into the adhesive layer confers upon it bactericidal properties in marked contrast to the non-antimicrobial adhesive, which contributed to bacterial proliferation when the wear time was ≥4 days. PMID:24722839

Antimicrobial activity of a novel adhesive containing chlorhexidine gluconate (CHG) against the resident microflora in human volunteers.

PubMed

Carty, Neal; Wibaux, Anne; Ward, Colleen; Paulson, Daryl S; Johnson, Peter

2014-08-01

To evaluate the antimicrobial activity of a new, transparent composite film dressing, whose adhesive contains chlorhexidine gluconate (CHG), against the native microflora present on human skin. CHG-containing adhesive film dressings and non-antimicrobial control film dressings were applied to the skin on the backs of healthy human volunteers without antiseptic preparation. Dressings were removed 1, 4 or 7 days after application. The bacterial populations underneath were measured by quantitative cultures (cylinder-scrub technique) and compared with one another as a function of time. The mean baseline microflora recovery was 3.24 log10 cfu/cm(2). The mean log reductions from baseline measured from underneath the CHG-containing dressings were 0.87, 0.78 and 1.30 log10 cfu/cm(2) on days 1, 4 and 7, respectively, compared with log reductions of 0.67, -0.87 and -1.29 log10 cfu/cm(2) from underneath the control film dressings. There was no significant difference between the log reductions of the two treatments on day 1, but on days 4 and 7 the log reduction associated with the CHG adhesive was significantly higher than that associated with the control adhesive. The adhesive containing CHG was associated with a sustained antimicrobial effect that was not present in the control. Incorporating the antimicrobial into the adhesive layer confers upon it bactericidal properties in marked contrast to the non-antimicrobial adhesive, which contributed to bacterial proliferation when the wear time was ≥4 days. © The Author 2014. Published by Oxford University Press on behalf of the British Society for Antimicrobial Chemotherapy.

Functionality of selected strains of moulds and yeasts from Vietnamese rice wine starters.

PubMed

Dung, N T P; Rombouts, F M; Nout, M J R

2006-06-01

The role of starch-degrading mycelial fungi, and the alcohol production and ethanol tolerance of the yeasts isolated from selected Vietnamese traditional rice wine starters were examined, and optimum conditions for these essential steps in rice wine fermentation were determined. Of pure isolates from Vietnamese rice wine starters, mould strains identified as Amylomyces rouxii, Amylomyces aff. rouxii, Rhizopus oligosporus and Rhizopus oryzae, were superior in starch degradation, glucose production and amyloglucosidase activity during the saccharification of purple glutinous rice. A. rouxii was able to produce up to 25%w/w glucose with an amyloglucosidase activity up to 0.6 Ug(-1) of fermented moulded mass. Five yeast isolates identified as Saccharomyces cerevisiae were selected for their superior alcohol productivity. They were able to deplete a relatively high initial percentage of glucose (20% w/v), forming 8.8% w/v ethanol. The ethanol tolerance of S. cerevisiae in challenge tests was 9-10% w/v, and 13.4% w/v as measured in fed-batch fermentations. Optimum conditions for the saccharification were: incubation for 2 d at 34 degrees C, of steamed rice inoculated with 5 log cfu g(-1); for the alcoholic fermentation 4 d at 28.3 degrees C, of saccharified rice liquid inoculated with 5.5 log cfu mL(-1).

The PK/PD Interactions of Doxycycline against Mycoplasma gallisepticum

PubMed Central

Zhang, Nan; Gu, Xiaoyan; Ye, Xiaomei; Wu, Xun; Zhang, Bingxu; Zhang, Longfei; Shen, Xiangguang; Jiang, Hongxia; Ding, Huanzhong

2016-01-01

Mycoplasma gallisepticum is one of the most important pathogens that cause chronic respiratory disease in chicken. This study investigated the antibacterial activity of doxycycline against M. gallisepticum strain S6. In static time–killing studies with constant antibiotic concentrations [0–64 minimum inhibitory concentration (MIC)], M. gallisepticum colonies were quantified and kill rates were calculated to estimate the drug effect. The half-life of doxycycline in chicken was 6.51 ± 0.63 h. An in vitro dynamic model (the drug concentrations are fluctuant) was also established and two half-lives of 6.51 and 12 h were simulated. The samples were collected for drug concentration determination and viable counting of M. gallisepticum. In static time–killing studies, doxycycline produced a maximum antimycoplasmal effect of 5.62log10 (CFU/mL) reduction and the maximum kill rate was 0.11 h−1. In the in vitro dynamic model, doxycycline had a mycoplasmacidal activity in the two regimens, and the maximum antimycoplasmal effects were 4.1 and 4.75log10 (CFU/mL) reduction, respectively. Furthermore, the cumulative percentage of time over a 48-h period that the drug concentration exceeds the MIC (%T > MIC) was the pharmacokinetic–pharmacodynamic index that best correlated with antimicrobial efficacy (R2 = 0.986, compared with 0.897 for the peak level divided by the MIC and 0.953 for the area under the concentration–time curve over 48 h divided by the MIC). The estimated %T > MIC values for 0log10 (CFU/mL) reduction, 2log10 (CFU/mL) reduction and 3log10 (CFU/mL) reduction were 32.48, 45.68, and 54.36%, respectively, during 48 h treatment period of doxycycline. In conclusion, doxycycline shows excellent effectiveness and time-dependent characteristics against M. gallisepticum strain S6 in vitro. Additionally, these results will guide optimal dosing strategies of doxycycline in M. gallisepticum infection. PMID:27199972

Inactivation of Bacteria S. aureus ATCC 25923 and S. Thyphimurium ATCC 14 028 Influence of UV-HPEF

NASA Astrophysics Data System (ADS)

Bakri, A.; Hariono, B.; Utami, M. M. D.; Sutrisno

2018-01-01

The research was objected to study the performance of the UV unit - HPEF in inactivating bacteria population of Gram-positive (S aureus ATCC 25923) and Gram-negative (S Thyphimurium ATCC 14028) inoculated in sterilized goat’s milk. UV pasteurization instrument employed three reactors constructed in series UV-C system at 10 W, 253.7 nm wavelength made in Kada (USA) Inc. with 1.8 J/cm2 dose per reactor. HPEF instrument used high pulsed electric field at 31.67 kV/cm, 15 Hz and goat’s milk rate at 4:32 ± 0.71 cc/second. Pathogenic bacteria was observed According to Indonesian National Standard 01-2782-1998. Inactivation rate of pathogenic bacteria ie S Thyphimurium ATCC 14028 and S. aureus ATCC 25923 was 0.28 and 0.19 log cycle or 6.35 and 4.34 log cfu/ml/hour, respectively; D value was 0.16 and 0.23 hour with k value was 14.62 and 10 hour-1 respectively.

Internalization of Listeria monocytogenes in Whole Avocado.

PubMed

Chen, Yi; Evans, Peter; Hammack, Thomas S; Brown, Eric W; Macarisin, Dumitru

2016-08-01

In recent years, tree fruits have emerged as a new concern for Listeria monocytogenes contamination. The objective of the current study was to evaluate the potential internalization of L. monocytogenes from the surface of avocados into the edible portions of the fruit during certain postharvest practices simulated in a laboratory setting. One set of intact avocados was spot inoculated with L. monocytogenes on the stem scar, and the second set was hydrocooled in water contaminated with L. monocytogenes. Under these experimental conditions, L. monocytogenes internalized into the avocado pulp through the stem or stem scar after both spot inoculation and hydrocooling. In avocados spot inoculated with 50, 130, 500, and 1,300 CFU per fruit, bacteria were detected in the edible portion adjacent to the stem scar within 15 days postinoculation during storage at 4°C. In avocados hydrocooled in water containing L. monocytogenes at 10(6) and 10(8) CFU/ml, bacteria reached the bottom end of the fruit, and the populations in the edible portion adjacent to the stem scar reached up to 5.90 to 7.19 log CFU/g within 10 to 15 days during storage at 4°C. Dye mixed with inoculum was useful for guiding subsequent sampling, but dye penetration patterns were not always consistent with bacterial penetration.

Control of spoiler Pseudomonas spp. on fresh cut vegetables by neutral electrolyzed water.

PubMed

Pinto, Loris; Ippolito, Antonio; Baruzzi, Federico

2015-09-01

In the present study, we evaluated the antimicrobial activity of neutral electrolyzed water (NEW) against 14 strains of spoilage Pseudomonas of fresh cut vegetables under cold storage. The NEW, produced from solutions of potassium and sodium chloride, and sodium bicarbonate developed up to 4000 mg/L of free chlorine, depending on the salt and relative concentration used. The antimicrobial effect of the NEW was evaluated against different bacterial strains at 10(5) cells/ml, with different combinations of free chlorine concentration/contact time; all concentrations above 100 mg/L, regardless of the salt used, were found to be bactericidal already after 2 min. When catalogna chicory and lettuce leaves were dipped for 5 min in diluted NEW, microbial loads of mesophilic bacteria and Enterobacteriaceae were reduced on average of 1.7 log cfu/g. In addition, when lettuce leaves were dipped in a cellular suspension of the spoiler Pseudomonas chicorii I3C strain, diluted NEW was able to reduce Pseudomonas population of about 1.0 log cfu/g. Thanks to its high antimicrobial activity against spoilage microorganisms, and low cost of operation, the application of cycles of electrolysis to the washing water looks as an effective tool in controlling fresh cut vegetable microbial spoilage contamination occurring during washing steps. Copyright © 2015 Elsevier Ltd. All rights reserved.

Antimicrobial effects of Lactobacillus plantarum and Lactobacillus acidophilus against multidrug-resistant enteroaggregative Escherichia coli.

PubMed

Kumar, Manesh; Dhaka, Pankaj; Vijay, Deepthi; Vergis, Jess; Mohan, Vysakh; Kumar, Ashok; Kurkure, Nitin V; Barbuddhe, Sukhadeo B; Malik, S V S; Rawool, Deepak B

2016-09-01

The in vitro and in vivo antimicrobial effects of Lactobacillus plantarum and Lactobacillus acidophilus were evaluated individually and synergistically against multidrug-resistant enteroaggregative Escherichia coli (MDR-EAEC). In vitro evaluation of each probiotic strain when co-cultured with MDR-EAEC isolates revealed a reduction in MDR-EAEC counts (eosin-methylene blue agar) in a dose- and time-dependent manner: probiotics at a dose rate of 10(10) CFU inhibited MDR-EAEC isolates at 72 h post-inoculation (PI), whereas at lower concentrations (10(8) and 10(9) CFU) MDR-EAEC isolates were inhibited at 96 h PI. The synergistic antimicrobial effect of both probiotic strains (each at 10(10) CFU) was highly significant (P < 0.01) and inhibited the growth of MDR-EAEC isolates at 24 h PI. For in vivo evaluation, weaned mice were fed orally with 10(7) CFU of MDR-EAEC. At Day 3 post-infection, treated mice were fed orally with the probiotic strains (each at 10(10) CFU). Compared with the control, post-treatment a significant (P < 0.01) reduction in MDR-EAEC counts was observed in faeces by Day 2 and in intestinal tissues of treated mice by Days 3 and 4 as evidenced by plate count (mean 2.71 log and 2.27 log, respectively) and real-time PCR (mean 1.62 log and 1.57 log, respectively) methods. Histopathologically, comparatively mild changes were observed in the ileum and colon from Days 3 to 5 post-treatment with probiotics; however, from Day 6 the changes were regenerative or normal. These observations suggest that these probiotic strains can serve as alternative therapeutics against MDR-EAEC-associated infections in humans and animals. Copyright © 2016 Elsevier B.V. and International Society of Chemotherapy. All rights reserved.

Microbial community, and biochemical and physiological properties of Korean traditional black raspberry (Robus coreanus Miquel) vinegar.

PubMed

Song, Nho-Eul; Cho, Sung-Ho; Baik, Sang-Ho

2016-08-01

The aim of this study was to elucidate the changes in microbial community and biochemical and physiological properties of traditional Muju black raspberry (Robus coreanus Miquel) vinegar (TMBV) during fermentation by culture-independent methods. During vinegar fermentation, ethanol produced up to 120 g L(-1) until day 35, with continuously increasing yeast concentration to a total of log 7.6 CFU mL(-1) . After day 35, acetic acid bacteria (AAB) concentrations rose to log 5.8 CFU mL(-1) until day 144. Denaturing gradient gel electrophoresis analysis showed that Saccharomyces cerevisiae was detected until day 87 of the fermentation, at which point Acetobacter pasteurianus gradually took over as the dominant species. Total sugar was reduced to 6.6 °Brix and total acidity produced up to 44 g L(-1) . In this study, we established the physicochemical analysis and growth dynamics of yeast and AAB during alcoholic and acetic acid fermentation of black raspberry by a traditional method. Overall, S. cerevisiae and A. pasteurianus species appeared to dominate the TMBV fermentation. In conclusion, this study demonstrated a suitable fermentation system for TMBV by the static surface method. © 2015 Society of Chemical Industry. © 2015 Society of Chemical Industry.

Competitiveness and antibacterial potential of bacteriocin-producing starter cultures in different types of fermented sausages.

PubMed

Ravyts, Frédéric; Barbuti, Silvana; Frustoli, Maria Angela; Parolari, Giovanni; Saccani, Giovanna; De Vuyst, Luc; Leroy, Frédéric

2008-09-01

Application of bacteriocin-producing starter cultures of lactic acid bacteria in fermented sausage production contributes to food safety. This is sometimes hampered by limited efficacy in situ and by uncertainty about strain dependency and universal applicability for different sausage types. In the present study, a promising antilisterial-bacteriocin producer, Lactobacillus sakei CTC 494, was applied as a coculture in addition to commercial fermentative starters in different types of dry-fermented sausages. The strain was successful in both Belgian-type sausage and Italian salami that were artificially contaminated with about 3.5 log CFU g(-1) of Listeria monocytogenes. After completion of the production process, this led to listerial reductions of up to 1.4 and 0.6 log CFU g(-1), respectively. In a control sausage, containing only the commercial fermentative starter, the reduction was limited to 0.8 log CFU g(-1) for the Belgian-type recipe, where pH decreased from 5.9 to 4.9, whereas an increase of 0.2 log CFU g(-1) was observed for Italian salami, in which the pH rose from 5.7 to 5.9 after an initial decrease to pH 5.3. In a Cacciatore recipe inoculated with 5.5 log CFU g(-1) of L. monocytogenes and in the presence of L. sakei CTC 494, there was a listerial reduction of 1.8 log CFU g(-1) at the end of the production process. This was superior to the effect obtained with the control sausage (0.8 log CFU g(-1)). Two commercial antilisterial cultures yielded reductions of 1.2 and 1.5 log CFU g(-1). Moreover, repetitive DNA sequence-based PCR fingerprinting demonstrated the competitive superiority of L. sakei CTC 494.

Microbial dynamics of indicator microorganisms on fresh tomatoes in the supply chain from Mexico to the USA.

PubMed

Zoellner, Claire; Venegas, Fabiola; Churey, John J; Dávila-Aviña, Jorge; Grohn, Yrjo T; García, Santos; Heredia, Norma; Worobo, Randy W

2016-12-05

Quality and safety of fresh produce are important to public health and maintaining commerce between Mexico and USA. While preventive practices can reduce risks of contamination and are generally successful, the variable environment of the supply chain of fresh produce can be suitable for introduction or proliferation of pathogenic microorganisms. As routine surveillance of these pathogens is not practical, indicator microorganisms are used to assess the sanitary conditions of production and handling environments. An opportunity exists to use indicators on fresh produce to measure how handling and transport from field to market may affect microbial populations that contribute to their quality or safety. The objective was to quantify indicator microorganisms on tomatoes sampled along the supply chain during the harvest year, in order to observe the levels and changes of populations at different locations. Roma tomatoes (n=475) were taken from the same lots (n=28) at four locations of the postharvest supply chain over five months: at arrival to and departure from the packinghouse in México, at the distribution center in Texas, and at retail in USA. Samples were analyzed individually for four microbial populations: aerobic plate count (APC), total coliforms (TC), generic Escherichia coli, and yeasts and molds (YM). APC population differed (p<0.05) from 1.9±1.1, 1.7±1.1, 2.3±1.1 and 3.5±1.4logCFU/g at postharvest, packing, distribution center and supermarket, respectively. TC populations were <1logCFU/g at postharvest, increased at packing (0.7±1.0logCFU/g), decreased in distribution (0.4±0.8logCFU/g) and increased in supermarkets (1.4±1.5logCFU/g). Generic E. coli was not identified from coliform populations in this supply chain. YM populations remained <1logCFU/g, with the exception of 1.1±1.3logCFU/g at supermarkets and tomatoes were not visibly spoiled. The levels reported from this pilot study demonstrated the dynamics within populations as influenced by time and conditions in one supply chain during a harvest year, while the large variances in some locations indicate opportunities for improvement. Overall, packinghouse and supermarket locations were identified as crucial points to control microbial safety risks. Copyright © 2016 Elsevier B.V. All rights reserved.

Modelling characteristics to predict Legionella contamination risk - Surveillance of drinking water plumbing systems and identification of risk areas.

PubMed

Völker, Sebastian; Schreiber, Christiane; Kistemann, Thomas

2016-01-01

For the surveillance of drinking water plumbing systems (DWPS) and the identification of risk factors, there is a need for an early estimation of the risk of Legionella contamination within a building, using efficient and assessable parameters to estimate hazards and to prioritize risks. The precision, accuracy and effectiveness of ways of estimating the risk of higher Legionella numbers (temperature, stagnation, pipe materials, etc.) have only rarely been empirically assessed in practice, although there is a broad consensus about the impact of these risk factors. We collected n = 807 drinking water samples from 9 buildings which had had Legionella spp. occurrences of >100 CFU/100mL within the last 12 months, and tested for Legionella spp., L. pneumophila, HPC 20°C and 36°C (culture-based). Each building was sampled for 6 months under standard operating conditions in the DWPS. We discovered high variability (up to 4 log(10) steps) in the presence of Legionella spp. (CFU/100 mL) within all buildings over a half year period as well as over the course of a day. Occurrences were significantly correlated with temperature, pipe length measures, and stagnation. Logistic regression modelling revealed three parameters (temperature after flushing until no significant changes in temperatures can be obtained, stagnation (low withdrawal, qualitatively assessed), pipe length proportion) to be the best predictors of Legionella contamination (>100 CFU/100 mL) at single outlets (precision = 66.7%; accuracy = 72.1%; F(0.5) score = 0.59). Copyright © 2015 Elsevier GmbH. All rights reserved.

Effects of NorA Inhibitors on In Vitro Antibacterial Activities and Postantibiotic Effects of Levofloxacin, Ciprofloxacin, and Norfloxacin in Genetically Related Strains of Staphylococcus aureus

PubMed Central

Aeschlimann, Jeffrey R.; Dresser, Linda D.; Kaatz, Glenn W.; Rybak, Michael J.

1999-01-01

NorA is a membrane-associated multidrug efflux protein that can decrease susceptibility to fluoroquinolones in Staphylococcus aureus. To determine the effect of NorA inhibition on the pharmacodynamics of fluoroquinolones, we evaluated the activities of levofloxacin, ciprofloxacin, and norfloxacin with and without various NorA inhibitors against three genetically related strains of S. aureus (SA 1199, the wild-type; SA 1199B, a NorA hyperproducer with a grlA mutation; and SA 1199-3, a strain that inducibly hyperproduces NorA) using susceptibility testing, time-kill curves, and postantibiotic effect (PAE) methods. Levofloxacin had the most potent activity against all three strains and was minimally affected by addition of NorA inhibitors. In contrast, reserpine, omeprazole, and lansoprazole produced 4-fold decreases in ciprofloxacin and norfloxacin MICs and MBCs for SA 1199 and 4- to 16-fold decreases for both SA 1199B and SA 1199-3. In time-kill experiments reserpine, omeprazole, or lansoprazole increased levofloxacin activity against SA 1199-3 alone by 2 log10 CFU/ml and increased norfloxacin and ciprofloxacin activities against all three strains by 0.5 to 4 log10 CFU/ml. Reserpine and omeprazole increased norfloxacin PAEs on SA 1199, SA 1199B, and SA 1199-3 from 0.9, 0.6, and 0.2 h to 2.5 to 4.5, 1.1 to 1.3, and 0.4 to 1.1 h, respectively; similar effects were observed with ciprofloxacin. Reserpine and omeprazole increased the levofloxacin PAE only on SA 1199B (from 1.6 to 5.0 and 3.1 h, respectively). In conclusion, the NorA inhibitors dramatically improved the activities of the more hydrophilic fluoroquinolones (norfloxacin and ciprofloxacin). These compounds may restore the activities of these fluoroquinolones against resistant strains of S. aureus or may potentially enhance their activities against sensitive strains. PMID:9925528

Hygiene intervention reduces contamination of weaning food in Bangladesh.

PubMed

Islam, Mohammad Sirajul; Mahmud, Zahid Hayat; Gope, Partha Sarathi; Zaman, Rokon Uz; Hossain, Zakir; Islam, Mohammad Shafiqul; Mondal, Dinesh; Sharker, Mohammad Abu Yushuf; Islam, Khairul; Jahan, Hasin; Bhuiya, Abbas; Endtz, Hubert P; Cravioto, Alejandro; Curtis, Valerie; Touré, Ousmane; Cairncross, Sandy

2013-03-01

This study was conducted to measure the impact of a hygiene intervention on the contamination of weaning food in Bangladesh. Sixty households were selected: 30 study and 30 control households. Samples of weaning food were collected from all the 60 households at baseline and examined for faecal coliforms (FC), faecal streptococci (FS) and Clostridium perfringens (CP) following standard procedures. After cooking, food samples were collected on three occasions before feeding. Following Hazard Analysis Critical Control Point (HACCP) procedures, critical control points were determined. The mothers in the 30 study households were then trained for 4 weeks in how to attain the control point conditions. Then, again the food samples were collected and analysed. At baseline, weaning foods from study and control households were heavily contaminated with FC and FS. The FC and FS counts were 1.84 log(10) and 1.92 log(10) colony-forming unit (cfu)/g, respectively, in the study households, and 0.86 log(10) and 1.33 log(10)  cfu/g, respectively, in the control households in the first feeding. After the intervention, the FC and FS counts in study households had dropped to 0.10 log(10) and 0.09 log(10)  cfu/g, respectively, a statistically significant reduction (P < 0.001). Monitoring the sustainability of the behaviour change after 3 months showed that the mothers were maintaining food hygiene. A hygiene intervention following the HACCP approach reduced the weaning food contamination significantly. Awareness building among mothers about weaning food hygiene could be an important intervention for preventing weaning food-related diarrhoea in Bangladesh. © 2012 Blackwell Publishing Ltd.

A signal-amplified electrochemical DNA biosensor incorporated with a colorimetric internal control for Vibrio cholerae detection using shelf-ready reagents.

PubMed

Low, Kim-Fatt; Zain, Zainiharyati Mohd; Yean, Chan Yean

2017-01-15

A novel enzyme/nanoparticle-based DNA biosensing platform with dual colorimetric/electrochemical approach has been developed for the sequence-specific detection of the bacterium Vibrio cholerae, the causative agent of acute diarrheal disease in cholera. This assay platform exploits the use of shelf-stable and ready-to-use (shelf-ready) reagents to greatly simplify the bioanalysis procedures, allowing the assay platform to be more amenable to point-of-care applications. To assure maximum diagnosis reliability, an internal control (IC) capable of providing instant validation of results was incorporated into the assay. The microbial target, single-stranded DNA amplified with asymmetric PCR, was quantitatively detected via electrochemical stripping analysis of gold nanoparticle-loaded latex microspheres as a signal-amplified hybridization tag, while the incorporated IC was analyzed using a simplified horseradish peroxidase enzyme-based colorimetric scheme by simple visual observation of enzymatic color development. The platform showed excellent diagnostic sensitivity and specificity (100%) when challenged with 145 clinical isolate-spiked fecal specimens. The limits of detection were 0.5ng/ml of genomic DNA and 10 colony-forming units (CFU)/ml of bacterial cells with dynamic ranges of 0-100ng/ml (R 2 =0.992) and log 10 (1-10 4 CFU/ml) (R 2 =0.9918), respectively. An accelerated stability test revealed that the assay reagents were stable at temperatures of 4-37°C, with an estimated ambient shelf life of 200 days. The versatility of the biosensing platform makes it easily adaptable for quantitative detection of other microbial pathogens. Copyright © 2016 Elsevier B.V. All rights reserved.

Live encapsulated Lactobacillus acidophilus cells in yogurt for therapeutic oral delivery: preparation and in vitro analysis of alginate-chitosan microcapsules.

PubMed

Urbanska, Aleksandra Malgorzata; Bhathena, Jasmine; Prakash, Satya

2007-09-01

Targeted delivery of live microencapsulated bacterial cells has strong potential for application in treating various diseases, including diarrhea, kidney failure, liver failure, and high cholesterol, among others. This study investigates the potential of microcapsules composed of two natural polymers, alginate and chitosan (AC), and the use of these artificial cells in yogurt for delivery of probiotic Lactobacillus acidophilus bacterial live cells. Results show that the integrity of AC microcapsules was preserved after 76 h of mechanical shaking in MRS broth and after 12 h and 24 h in simulated gastric and intestinal fluids. Using an in vitro computer-controlled simulated human gastrointestinal (GI) model, we found 8.37 log CFU/mL of viable bacterial cells were present after 120 min of gastric exposure and 7.96 log CFU/mL after 360 min of intestinal exposure. In addition, AC microcapsules composed of chitosan 10 and 100 at various concentrations were subjected to 4-week storage in 2% milk fat yogurt or 0.85% physiological solution. It was found that 9.37 log CFU/mL of cells encapsulated with chitosan 10 and 8.24 log CFU/mL of cells encapsulated with chitosan 100 were alive after 4 weeks. The AC capsule composed of 0.5% chitosan 10 provided the highest bacterial survival of 9.11 log CFU/mL after 4 weeks. Finally, an investigation of bacterial viability over 72 h in different pH buffers yielded highest survival of 6.34 log CFU/mL and 10.34 log CFU/mL at pH 8 for free and AC-encapsulated cells, respectively. We conclude from these findings that encapsulation allows delivery of a higher number of bacteria to desired targets in the GI tract and that microcapsules containing bacterial cells are good candidates for oral artificial cells for bacterial cell therapy.

Comparison of the ActiDes-Blue and CARELA HYDRO-DES technology for the sanitation of contaminated cooling water systems in dental units.

PubMed

Kramer, Axel; Lemanski, Sandra; Demond, Kathleen; Assadian, Ojan

2012-01-01

The hygienic-microbiological control of 6 dental units being in use for the past 16 years revealed a significantly increased microbial contamination of their cooling water system. In order to comply with the requirements of the drinking water directive ("Trinkwasserverordnung"), the commercially available production system ActiDes, producing on-site ActiDes-Blue which is based on hypochlorous acid (HOCl) and generated by anodic oxidation, was investigated. Water samples from the 6 contaminated dental units were examined for the total number of colony forming units (cfu), contamination with molds, L. pneumophila and P. aeruginosa. The control period for the total colony count was 4 weeks (8 samples/unit). The subsequent application phase of the ActiDes-Blue procedure was 6 months (31 samples/unit). Additionally, the redox potential and the pH value were measured.Futhermore, the decontamination agent CARELA HYDRO-DES, a two component agent based on H(2)O(2) with the addition of a mixture of sodium hydrogen sulphate and sulphuric acid in an aqueous solution effective at 0.1% and higher, was applied in a unit that had been put out of service for a month before. Before application, the system was first filled with a 5% solution of the alkaline pre-cleaning agent CARELA Solvent for bacterial slime; the system was left with this solution for 1 h. The pre-cleaning agent was then completely displaced from the system with tap water and a decontaminating solution of 5% CARELA HYDRO-DES and left in place for 1 h. Drinking water quality level was reached only twice during the control phase. The average values of the dental units ranged between 3,633 CFU/ml and 29,417 c/ml. During the application phase, drinking water level could be achieved in 11 water samples. In another 6 water samples a total colony count of <150 cfu/ml was reached. The average values for the dental units' total colony count ranged between 529 cfu/ml and 87,450 cfu/ml. No significant differences between the control phase and the action phase could be demonstrated. During the control phase, contamination of the water samples with a mold was noticed so that examinations for molds were carried out beyond the scope of the drinking water directive. For this parameter as well, no significant differences between the phases of the study could be shown.The Legionella load of the dental units was low. L. pneumophila were yielded in only 4 out of 130 water samples. During the control phase, twice colony counts at 50 cfu/1,000 ml and 110 cfu/1,000 ml were measured. During the action phase, counts with Legionella spp. could be measured at 5 cfu/1,000 ml for one unit only. Also, with 1-10 cfu/100 ml, the P. aeruginosa contamination was low. During the application phase, it ranged between 0-7 cfu/100 ml.Redox potential and pH value showed a slight decrease during the application phase.Before treatment with CARELA Solvent and CARELA HYDRO-DES, the initial contamination of the total count of bacterial colonies was 1,432 cfu/ml at 22°C and 846 cfu/ml at 36°C as well as >1,000 cfu/100 ml for molds. 1 h after the decontamination, no bacteria and molds could be detected in 1,000 ml of tap water. Despite the fact that the unit was not used any longer, after 7 d the bacterial colony count was 3 cfu/ml at 22°C and 2 cfu/ml at 36°C while molds could not be detected. Even after a rest time of 14 d only 167 cfu/ml or 42 cfu/ml could be yielded. Molds were further not cultivable. A material damage could not be observed. Pertaining to the ActiDes technology's effectiveness, it has to be pointed out that the dental units investigated were those used for dental students' teaching and therefore were clearly less frequently used than clinically used units in a dental practice. This resulted in distinctly longer stagnation periods which favored formation of biofilms. In summary, the ActiDes technology and ActiDes-Blue showed not to be sufficiently effective for the sanitation of contaminated water reservoirs in dental units under aggravated conditions of repeated and longer periods of non-use in connection with longer water stagnation periods. In comparison, the biofilm was sustainably eliminated through the combined application of CARELA(®) Solvent for Bacterial Slime with subsequent decontamination using CARELA(®) HYDRO-DES.

The effect of high-level chlorine carcass drench on the recovery of Salmonella and enumeration of bacteria from broiler carcasses.

PubMed

Bartenfeld, L N; Fletcher, D L; Northcutt, J K; Bourassa, D V; Cox, N A; Buhr, R J

2014-11-01

A study was conducted to determine the bacteriological effect of exposing processed broiler carcasses to a high (10-fold increase) concentration chlorinated drench. During each of 6 replicate trials, eviscerated prechill carcasses were obtained from a commercial processing plant and chlorine-treated carcasses were subjected to a 1-min drench in 500 mL of a 500 mg/kg chlorine solution (sodium hypochlorite). Water-drenched carcasses were treated the same way except water was used in place of chlorinated water drench. Control carcasses were not drenched. All carcasses were then subjected to a whole carcass rinse (WCR) in 450 mL of buffered peptone water, from which 50 mL of the rinsate was removed for enumeration of total aerobic bacteria (APC), Escherichia coli, and total coliforms (TC). The entire carcass was then incubated 24 h at 37°C (whole carcass enrichment, WCE) for recovery of Salmonella. Levels of bacteria recovered from WCR were lower by 0.6 log10 cfu/mL for APC, 0.8 for E. coli, and 0.9 for TC when carcasses were drenched with water compared with undrenched control levels. Similarly, the levels of bacteria recovered from WCR were further lower by 1.0 log10 cfu/mL for APC, 0.5 for E. coli, and 0.5 for TC, when carcasses were drenched with 500 mg/kg of chlorine compared with water. However, there was no significant difference (P > 0.05) in prevalence of Salmonella among the treatments (29% positive for control, 26% positive for water, 38% positive for chlorinated). These results indicate that drenching eviscerated carcasses with water or chlorinated water at 500 mg/kg significantly, but minimally, reduces the numbers of APC, E. coli, and TC bacteria recovered compared with undrenched carcasses. However, neither drenching carcasses with water or high chlorine had an effect on the prevalence of Salmonella that remain with the carcass as determined by WCE. The results of this study confirms the importance of maintaining and replenishing free chlorine for optimal antimicrobial activity, because chlorine at 500 mg/kg was rapidly used within 1 min of exposure to the carcass to <10 mg/kg. ©2014 Poultry Science Association Inc.

Use of an Electrostatic Spraying System or the Sprayed Lethality in Container Method To Deliver Antimicrobial Agents onto the Surface of Beef Subprimals To Control Shiga Toxin-Producing Escherichia coli.

PubMed

Stella, J Max; Luchansky, John B; Miller, Kelsey; Shoyer, Bradley A; Shane, Laura E; McGeary, Lianna; Osoria, Manuela; Stahler, Laura J; Sevart, Nicholas J; Phebus, Randall K; Thippareddi, Harshavardhan; Porto-Fett, Anna C S

2017-08-01

The efficacy of an electrostatic spraying system (ESS) and/or the sprayed lethality in container (SLIC) method to deliver antimicrobial agents onto the surface of beef subprimals to reduce levels of Shiga toxin-producing Escherichia coli (STEC) was evaluated. Beef subprimals were surface inoculated (lean side; ca. 5.8 log CFU per subprimal) with 2 mL of an eight-strain cocktail comprising single strains of rifampin-resistant (100 μg/mL) STEC (O26:H11, O45:H2, O103:H2, O104:H4, O111:H - , O121:H19, O145:NM, and O157:H7). Next, inoculated subprimals were surface treated with lauric arginate (LAE; 1%), peroxyacetic acid (PAA; 0.025%), or cetylpyridinium chloride (CPC; 0.4%) by passing each subprimal, with the inoculated lean side facing upward, through an ESS cabinet or via SLIC. Subprimals were then vacuum packaged and stored at 4°C. One set of subprimals was sampled after an additional 2 h, 3 days, or 7 days of refrigerated storage, whereas another set was retreated via SLIC after 3 days of storage with a different one of the three antimicrobial agents (e.g., a subprimal treated with LAE on day 0 was then treated with PAA or CPE on day 3). Retreated subprimals were sampled after 2 h or 4 days of additional storage at 4°C. A single initial application of LAE, PAA, or CPC via ESS or SLIC resulted in STEC reductions of ca. 0.3 to 1.3 log CFU per subprimal after 7 days of storage. However, when subprimals were initially treated with LAE, PAA, or CPC via ESS or SLIC and then separately retreated with a different one of these antimicrobial agents via SLIC on day 3, additional STEC reductions of 0.4 to 1.0 log CFU per subprimal were observed after an additional 4 days of storage. Application of LAE, PAA, or CPC, either alone or in combination, via ESS or SLIC is effective for reducing low levels (ca. 0.3 to 1.6 log CFU) of STEC that may be naturally present on the surface of beef subprimals.

Eating oysters without risk of vibriosis: application of a bacteriophage against Vibrio parahaemolyticus in oysters.

PubMed

Jun, Jin Woo; Kim, Hyoun Joong; Yun, Sae Kil; Chai, Ji Young; Park, Se Chang

2014-10-01

Vibrio parahaemolyticus is a major cause of foodborne illness and related with the consumption of raw contaminated seafood, especially oysters. To evaluate the effectiveness of various applications of a bacteriophage (phage), pVp-1, against a multiple-antibiotic-resistant V. parahaemolyticus pandemic strain (CRS 09-17), we designed artificial contamination models that are most likely to be encountered during oyster processing. When live oysters were treated with bath immersion with pVp-1 after CRS 09-17 challenge, the growth of bacterial strain was significantly reduced. After 72h of phage application with bath immersion, bacterial growth reduction was observed to be 8.9×10(6)CFU/ml (control group) to 1.4×10CFU/ml (treatment group). When pVp-1 was surface-applied on the flesh of oysters after CRS 09-17 inoculation, bacterial growth was properly inhibited. After 12h of phage application on the surface of oysters, bacterial growth inhibition was revealed to be 1.44×10(6)CFU/ml (control group) to 1.94CFU/ml (treatment group). This is the first report, to the best of our knowledge, of oyster surface-application of a phage against a multiple-antibiotic-resistant V. parahaemolyticus pandemic strain, and our successful phage application to various situations emphasizes the potential use of the phage to avoid V. parahaemolyticus infection from aquaculture to consumption. Copyright © 2014 Elsevier B.V. All rights reserved.

Effects of ultraviolet light emitting diodes (LEDs) on microbial and enzyme inactivation of apple juice.

PubMed

Akgün, Merve Pelvan; Ünlütürk, Sevcan

2017-11-02

In this study, the effects of Ultraviolet light-emitting diodes (UV-LEDs) on the inactivation of E. coli K12 (ATCC 25253), an indicator organism of E. coli O157:H7, and polyphneoloxidase (PPO) in cloudy apple juice (CAJ) were investigated. The clear (AJ) and cloudy apple juice were exposed to UV rays for 40min by using a UV device composed of four UV-LEDs with peak emissions at 254 and 280nm and coupled emissions as follows: 254/365, 254/405, 280/365, 280/405 and 254/280/365/405nm. UV-LEDs at 254nm achieved 1.6±0.1 log 10 CFU/mL inactivation of E. coli K12 at UV dose of 707.2mJ/cm 2 . The highest inactivation of E. coli K12 (2.0±0.1log 10 CFU/mL and 2.0±0.4log 10 CFU/mL) was achieved when the cloudy apple juice was treated with both 280nm and 280/365nm UV-LEDs. For clear apple juice the highest inactivation 4.4log 10 CFU/mL obtained for E. coli K12 was achieved using 4 lamps emitting light at 280nm for 40min exposure time. For the same treatment time, the experiments using a combination of lamps emitting light at 280 and 365nm (2lamp/2lamp) were resulted in 3.9±0.2log 10 CFU/mL reductions. UV-A and UV-C rays in combination showed a better inactivation effect on PPO than UV-C rays used separately. Residual activity of PPO in CAJ was reduced to 32.58% when treated with UV-LED in combination of UV-C (280nm) and UV-A (365nm) rays. Additionally, the total color change (ΔE) of CAJ subjected to combined UV-LED irradiation at 280/365nm was the lowest compared to other studied processing conditions. This study provides key implications for the future application of UV-LEDs to fruit juice pasteurization. Copyright © 2017 Elsevier B.V. All rights reserved.

The use and performance of BioSand filters in the Artibonite Valley of Haiti: a field study of 107 households.

PubMed

Duke, W F; Nordin, R N; Baker, D; Mazumder, A

2006-01-01

Approximately one billion people world-wide lack access to adequate amounts of safe water. Most are in developing countries, especially in rapidly expanding urban fringes, poor rural areas, and indigenous communities. In February and March 2005, a field study of 107 households was conducted to evaluate the use and performance of the Manz BioSand filter in the Artibonite Valley of Haiti. Approximately 2000 filters had been installed in this area over the preceding 5 years by the staff in Community Development at Hospital Albert Schweitzer, Deschappelle, Haiti. Interviews, observations, and water samplings were carried-out by two teams of Haitian enumerators, each consisting of a nurse and a filter technician. Water analyses were performed by Haitian lab technicians using the membrane filtration method to determine Escherichia coli counts. The enumerators and the lab technicians completed a 2 week training program before beginning the study; they worked under the direct supervision of the primary investigator. Laboratory quality was monitored by running 10% blank and 10% duplicate samples. The households contained an average of 5.4 persons. Filters had been in use for an average of 2.5 years, and participants were generally satisfied with their filter's performance. Shallow, hand-dug wells provided the only source of water for 61% of the households, with 26% using water piped from springs or deep wells, and 13% having access to both. Only 3% had plumbing in their homes. Source water from shallow wells contained an average of 234 E. coli cfu/100 mL. Piped sources averaged 195 E. coli cfu/100 mL. Of the source water samples 26% contained 0-10 E. coli cfu/100 mL. Of the filtered water samples 97% contained 0-10 E. coli cfu/100 mL (80% with 0 cfu/100 mL, and 17% with 1-10 cfu/100 mL). Overall bacterial removal efficiency for the filters was calculated to be 98.5%. Turbidity decreased from an average of 6.2 NTU in source water samples to 0.9 NTU in the filtered water. None of the households treated the water after filtering; 91% used the filtered water only for drinking. No problems related to filter construction were observed; 13% were found to have significantly decreased flow rates (all restored by cleaning the filter). Recontamination was found to occur, with only 3% of the samples from the filters' spouts containing >10 E. coli cfu/100 mL and 22% of the stored filtered water samples at point-of-use containing >10 cfu/100 mL. The Manz BioSand filters are an attractive option for supplying water treatment to family units in rural areas of poorly developed countries.

Evaluation of a lytic bacteriophage, Φ st1, for biocontrol of Salmonella enterica serovar Typhimurium in chickens.

PubMed

Wong, Chuan Loo; Sieo, Chin Chin; Tan, Wen Siang; Abdullah, Norhani; Hair-Bejo, Mohd; Abu, Jalila; Ho, Yin Wan

2014-02-17

In this study, a Salmonella Typhimurium lytic bacteriophage, Φ st1, which was isolated from chicken faecal material, was evaluated as a candidate for biocontrol of Salmonella in chickens. The morphology of Φ st1 showed strong resemblance to members of the Siphoviridae family. Φ st1 was observed to be a DNA phage with an estimated genome size of 121 kbp. It was found to be able to infect S. Typhimurium and S. Hadar, with a stronger lytic activity against the former. Subsequent characterisation of Φ st1 against S. Typhimurium showed that Φ st1 has a latent period of 40 min with an average burst size of 22 particles per infective centre. Approximately 86.1% of the phage adsorbed to the host cells within the initial 5 min of infection. At the optimum multiplicity of infection (MOI) (0.1), the highest reduction rate of S. Typhimurium (6.6 log₁₀ CFU/ml) and increment in phage titre (3.8 log₁₀ PFU/ml) was observed. Φ st1 produced adsorption rates of 88.4-92.2% at pH7-9 and demonstrated the highest bacteria reduction (6.6 log₁₀ CFU/ml) at pH9. Φ st1 also showed an insignificant different (P>0.05) reduction rate of host cells at 37 °C (6.4 log₁₀ CFU/ml) and 42 °C (6.0 log₁₀ CFU/ml). The in vivo study using Φ st1 showed that intracloacal inoculation of ~10¹² PFU/ml of the phage in the chickens challenged with ~10¹⁰ CFU/ml of S. Typhimurium was able to reduce (P<0.05) the S. Typhimurium more rapidly than the untreated group. The Salmonella count reduced to 2.9 log₁₀ CFU/ml within 6h of post-challenge and S. Typhimurium was not detected at and after 24h of post-challenge. Reduction of Salmonella count in visceral organs was also observed at 6h post-challenge. Approximately 1.6 log₁₀ FU/ml Φ st1 was found to persist in the caecal wall of the chicks at 72 h of post-challenge. The present study indicated that Φ st1 may serve as a potential biocontrol agent to reduce the Salmonella count in caecal content of chickens. Copyright © 2013 Elsevier B.V. All rights reserved.

Evaluation of Antimicrobial Activities of Sequential Spray Applications of Decontamination Treatments on Chicken Carcasses

PubMed Central

Benli, Hakan; Sanchez-Plata, Marcos X.; Ilhak, Osman Irfan; Núñez De González, Maryuri T.; Keeton, Jimmy T.

2015-01-01

The objective of this study was to evaluate the effects of sequential applications of ɛ-polylysine (EPL) or lauramide arginine ethyl ester (LAE) sprays followed by an acidic calcium sulfate (ACS) spray on inoculated chicken carcasses to reduce Salmonella (Salmonella enterica serovars including Salmonella typhimurium and Salmonella enteritidis) contamination during 6 days of storage (4.4°C). Secondly, reductions of the resident microflora were studied on uninoculated chicken carcasses following the sequential application of the treatments, chilling and 10 days of storage at 4.4°C. The treatment of Salmonella inoculated carcasses with 300 mg/L EPL followed by 30% ACS (EPL300-ACS30) sprays reduced Salmonella counts initially by 1.5 log cfu/mL and then by 1.2 log cfu/mL (p<0.05) following 6 days of storage at 4.4°C. Likewise, 200 mg/L LAE followed by 30% ACS (LAE200-ACS30) treatment reduced initial Salmonella counts on poultry carcasses by 1.8, 1.4 and 1.8 log cfu/mL (p<0.05), respectively, after 0, 3, and 6 days storage. Immediately after the treatments, EPL300-ACS30 and LAE200-ACS30 both reduced Escherichia coli counts significantly by 2.6 and 2.9 log cfu/mL, respectively. EPL300-ACS30 and LAE200-ASC30 were effective in lowering psychrotroph counts by 1 log cfu/mL on day 10 when compared to the control and distilled water treatments. This study demonstrated that EPL300-ACS30 and LAE200-ACS30 were effective in reducing Salmonella on inoculated chicken carcasses both after treatment and during the storage at 4.4°C for up to 6 days. In addition, reductions in psychrotroph counts indicated that these treatments might have the potential to increase the shelf-life of poultry carcasses. PMID:25656180

Microencapsulation of Lactobacillus plantarum spp in an alginate matrix coated with whey proteins.

PubMed

Gbassi, Gildas Komenan; Vandamme, Thierry; Ennahar, Saïd; Marchioni, Eric

2009-01-31

Whey proteins were used as a coating material to improve encapsulation of Lactobacillus plantarum strains in calcium alginate beads. L. plantarum 299v, L. plantarum 800 and L. plantarum CIP A159 were used in this study. Inactivation experiments were carried out in simulated gastric fluid (SGF) and simulated intestinal fluid (SIF). Cross-sections of freeze-dried beads revealed the random distribution of bacteria throughout the alginate network. From an initial count of 10.04+/-0.01 log(10) CFU g(-1) for L. plantarum 299v, 10.12+/-0.04 for L. plantarum CIP A159 and 10.03+/-0.01 for L. plantarum 800, bacteria in coated beads and incubated in SGF (37 degrees C, 60 min) showed a better survival for L. plantarum 299v, L. plantarum CIP A159 and L. plantarum 800 (respectively 7.76+/-0.12, 6.67+/-0.08 and 5.81+/-0.25 log(10) CFU g(-1)) when compared to uncoated beads (2.19+/-0.09, 1.89+/-0.09 and 1.65+/-0.10 log(10) CFU g(-1)) (p<0.05). Only bacteria in the coated beads survived in the SIF medium (37 degrees C, 180 min) after SGF treatment. This preliminary work showed that whey proteins are a convenient, cheap and efficient material for coating alginate beads loaded with bacteria.

Inactivation of Bacillus anthracis Spores during Laboratory-Scale Composting of Feedlot Cattle Manure

PubMed Central

Xu, Shanwei; Harvey, Amanda; Barbieri, Ruth; Reuter, Tim; Stanford, Kim; Amoako, Kingsley K.; Selinger, Leonard B.; McAllister, Tim A.

2016-01-01

Anthrax outbreaks in livestock have social, economic and health implications, altering farmer’s livelihoods, impacting trade and posing a zoonotic risk. Our study investigated the survival of Bacillus thuringiensis and B. anthracis spores sporulated at 15, 20, or 37°C, over 33 days of composting. Spores (∼7.5 log10 CFU g-1) were mixed with manure and composted in laboratory scale composters. After 15 days, the compost was mixed and returned to the composter for a second cycle. Temperatures peaked at 71°C on day 2 and remained ≥55°C for an average of 7 days in the first cycle, but did not exceed 55°C in the second. For B. thuringiensis, spores generated at 15 and 21°C exhibited reduced (P < 0.05) viability of 2.7 and 2.6 log10 CFU g-1 respectively, as compared to a 0.6 log10 CFU g-1 reduction for those generated at 37°C. For B. anthracis, sporulation temperature did not impact spore survival as there was a 2.5, 2.2, and 2.8 log10 CFU g-1 reduction after composting for spores generated at 15, 21, and 37°C, respectively. For both species, spore viability declined more rapidly (P < 0.05) in the first as compared to the second composting cycle. Our findings suggest that the duration of thermophilic exposure (≥55°C) is the main factor influencing survival of B. anthracis spores in compost. As sporulation temperature did not influence survival of B. anthracis, composting may lower the viability of spores associated with carcasses infected with B. anthracis over a range of sporulation temperatures. PMID:27303388

Enhanced Reduction of Helicobacter pylori Load in Precolonized Mice Treated with Combined Famotidine and Urease-Binding Polysaccharides

PubMed Central

Icatlo, Faustino C.; Kimura, Nobutake; Goshima, Hideo; Kodama, Yoshikatsu

2000-01-01

The present study investigated the effect of a model urease-binding polysaccharide in combination with a histamine H2 receptor antagonist on Helicobacter pylori colonization in vivo. Euthymic hairless mice were treated daily with dextran sulfate via drinking water and/or famotidine via intragastric gavage starting at 1 week postchallenge with a CagA+ VacA+ (type 1) strain of H. pylori. Treatment of precolonized mice for 2 weeks with dextran sulfate combined with famotidine yielded a group mean bacterial load (per 100 mg of gastric tissue) of log10 1.04 CFU, which was significantly lower than those of the famotidine (log10 3.35 CFU, P < 0.01) and dextran sulfate (log10 2.45 CFU, P < 0.05) monotherapy groups and the infected nontreated group (log10 3.64 CFU, P < 0.01). Eradication was achieved after 2 weeks of treatment in 50% or more of the test mice using drug combinations (1 or 2 weeks of famotidine plus 2 weeks of dextran sulfate) versus none in the monotherapy and positive control groups. The enhanced activity of the drug combination may be related to the daily pattern of transient acid suppression by famotidine inducing periodic bacterial convergence to superficial mucus sites penetrated by dextran sulfate from the lumen. Increased urease-dextran sulfate avidity was observed in vitro in the presence of famotidine and may partly account for the enhanced activity. With potential utility in abbreviating treatment time and eradication of antibiotic-resistant strains, the use of urease-targeted polysaccharides concurrently with a gastric acid inhibitor warrants consideration as an additional component of the standard multidrug chemotherapy of H. pylori infection. PMID:10952600

Effect of pH on the survival of Listeria innocua in calcium ascorbate solutions and on quality of fresh-cut apples.

PubMed

Karaibrahimoglu, Yildiz; Fan, Xuetong; Sapers, Gerald M; Sokorai, Kimberly

2004-04-01

Fresh-cut apple slices were dipped in calcium ascorbate (CaA) solution at pH values ranging from 2.5 to 7.0 to inhibit browning. After treatment, the cut apples were stored at 4 and 10 degrees C for up to 21 days. Color and texture of the apples were determined on days 1, 14, and 21. In a separate experiement, the pH of CaA solution was adjusted with acetic acid to six different pH levels, and the solution was inoculated with Listeria innocua. The survival of the bacterium and the stability of CaA were determined at 0, 20, and 96 h. The cut apples maintained fresh quality when the pH of the CaA solution was above 4.5, but slight discoloration of apple slices dipped in pH 4.5 solution was observed after 14 days at 10 degrees C. At pH 5.0, the CaA dip maintained the quality of the apples at both temperatures for at least 21 days. The L. innocua population was reduced by 4 to 5 log CFU/ml at pH 4.5 after 96 h. At pH 5, the bacterial population in the CaA solution was reduced by approximately 2 log CFU/ml during the same period. The CaA solution was stable at pH 5 for at least 96 h. Reduction of the pH to between 4.5 and 5.0 might reduce the risk of foodborne illness due to consumption of fresh-cut apples treated with a CaA solution contaminated with Listeria.

Effect of species, breed, and age on bacterial load in bovine and bubaline semen

PubMed Central

Sannat, Chandrahas; Nair, Ajit; Sahu, S. B.; Sahasrabudhe, S. A.; Kumar, Ashish; Gupta, Amit Kumar; Shende, R. K.

2015-01-01

Aim: The present study was conducted to investigate the effect of species, breed and age on bacterial load in fresh and frozen semen of Cattle and Buffalo bull. Materials and Methods: Present study covered 56 cow and 10 buffalo bulls stationed at Central Semen Station Anjora, Durg (Chhattisgarh). Impact of breeds on bacterial load in semen was assessed using six breeds of cattle viz. Sahiwal, Gir, Red Sindhi, Tharparkar, Jersey and Holstein Friesian (HF) cross. Cow bulls were categorized into four different groups based on their age (<4 years, 4-5 years, 5-6 years and > 6 years) to study variation among age groups. Bacterial load was measured in fresh and frozen semen samples from these bulls using the standard plate count (SPC) method and count was expressed as colony forming unit (CFU) per ml of semen. Results: Higher bacterial load was reported in fresh (2.36 × 104 ± 1943 CFU/ml) and frozen (1.00 × 10 ± 90 CFU/ml) semen of cow bulls as compared to buffalo bulls (1.95 × 104 ± 2882 and 7.75 × 102 ± 160 CFU/ml in fresh and frozen semen, respectively). Jersey bull showed significantly higher bacterial count (p < 0.05) both in fresh (4.07 × 104 ± 13927 CFU/ml) and frozen (1.92 × 103 ± 178 CFU/ml) semen followed by HF cross, Sahiwal, Gir, Red Sindhi and Tharparkar bull. Bulls aged < 4 years and more than 6 years yielded increased bacterial load in their semen. Although a minor variation was reported between species and among age groups, no significant differences were measured. Conclusion: Bacterial load in semen did not differ significantly between species and age groups; however significant variation was reported among different breeds. Bulls of Jersey breed showed significantly higher bacterial load in semen as compared to the crossbred and indigenous bull. PMID:27047115

Salmonella enterica growth and biofilm formation in flesh and peel cantaloupe extracts on four food-contact surfaces.

PubMed

De Abrew Abeysundara, Piumi; Dhowlaghar, Nitin; Nannapaneni, Ramakrishna; Schilling, Mark W; Mahmoud, Barakat; Sharma, Chander S; Ma, Din-Pow

2018-05-04

Salmonella enterica is responsible for the highest number of foodborne disease outbreaks pertaining to cantaloupe industry. The objective of this study was to examine the growth and biofilm formation by outbreak strains of S. enterica ser. Poona (S. Poona), S. enterica ser. Stanley (S. Stanley) and S. enterica ser. Montevideo (S. Montevideo) on different food-contact processing surfaces in cantaloupe flesh and peel extracts at 22 °C and 10 °C. The generation time of all S. enterica strains tested was shorter in the high concentration (50 mg/ml) of cantaloupe extract and high temperature. In 50 mg/ml of cantaloupe flesh or peel extract, the populations of S. enterica were increased by 5 log CFU/ml in 24 h at 22 °C and 1 log CFU/ml in 72 h at 10 °C. In 2 mg/ml of cantaloupe flesh or peel extracts, the populations of S. enterica were increased by 3.5 log CFU/ml in 56 h at 22 °C, but there were no changes in 72 h at 10 °C. The biofilm production of S. enterica was greater at 50 mg/ml of cantaloupe extract and 22 °C, but no major differences (P ≥ 0.05) were found among the strains tested. In 50 mg/ml cantaloupe extract, S. enterica produced 5-6 log CFU/cm 2 biofilm in 4-7 d at 22 °C and approximately 3.5-4 log CFU/cm 2 in 7 d at 10 °C. In 2 mg/ml of cantaloupe extract, S. enterica produced 4-4.5 log CFU/cm 2 biofilms in 4-7 d at 22 °C and 3 log CFU/cm 2 in 7 d at 10 °C. Biofilm formation by S. Poona (01A4754) was lowest on buna-n rubber compared to stainless steel, polyethylene and polyurethane surfaces under the majority of conditions tested. Overall, these findings show that S. enterica strains can grow rapidly and form biofilms on different cantaloupe processing surfaces in the presence of low concentrations of cantaloupe flesh or peel extracts. Copyright © 2018 Elsevier B.V. All rights reserved.

Heterotrophic bacteria in an air-handling system.

PubMed Central

Hugenholtz, P; Fuerst, J A

1992-01-01

Heterotrophic bacteria from structural surfaces, drain pan water, and the airstream of a well-maintained air-handling system with no reported building-related illness were enumerated. Visually the system appeared clean, but large populations of bacteria were found on the fin surface of the supply-side cooling coils (10(5) to 10(6) CFU cm-2), in drain pan water (10(5) to 10(7) CFU ml-1), and in the sump water of the evaporative condenser (10(5) CFU ml-1). Representative bacterial colony types recovered from heterotrophic plate count cultures on R2A medium were identified to the genus level. Budding bacteria belonging to the genus Blastobacter dominated the supply surface of the coil fins, the drain pan water, and the postcoil air. These data and independent scanning electron microscopy indicated that a resident population of predominantly Blastobacter bacteria was present as a biofilm on the supply-side cooling coil fins. Images PMID:1476435

Heterotrophic bacteria in an air-handling system.

PubMed

Hugenholtz, P; Fuerst, J A

1992-12-01

Heterotrophic bacteria from structural surfaces, drain pan water, and the airstream of a well-maintained air-handling system with no reported building-related illness were enumerated. Visually the system appeared clean, but large populations of bacteria were found on the fin surface of the supply-side cooling coils (10(5) to 10(6) CFU cm-2), in drain pan water (10(5) to 10(7) CFU ml-1), and in the sump water of the evaporative condenser (10(5) CFU ml-1). Representative bacterial colony types recovered from heterotrophic plate count cultures on R2A medium were identified to the genus level. Budding bacteria belonging to the genus Blastobacter dominated the supply surface of the coil fins, the drain pan water, and the postcoil air. These data and independent scanning electron microscopy indicated that a resident population of predominantly Blastobacter bacteria was present as a biofilm on the supply-side cooling coil fins.

Kinetic Behavior of Escherichia coli on Various Cheeses under Constant and Dynamic Temperature.

PubMed

Kim, K; Lee, H; Gwak, E; Yoon, Y

2014-07-01

In this study, we developed kinetic models to predict the growth of pathogenic Escherichia coli on cheeses during storage at constant and changing temperatures. A five-strain mixture of pathogenic E. coli was inoculated onto natural cheeses (Brie and Camembert) and processed cheeses (sliced Mozzarella and sliced Cheddar) at 3 to 4 log CFU/g. The inoculated cheeses were stored at 4, 10, 15, 25, and 30°C for 1 to 320 h, with a different storage time being used for each temperature. Total bacteria and E. coli cells were enumerated on tryptic soy agar and MacConkey sorbitol agar, respectively. E. coli growth data were fitted to the Baranyi model to calculate the maximum specific growth rate (μ max; log CFU/g/h), lag phase duration (LPD; h), lower asymptote (log CFU/g), and upper asymptote (log CFU/g). The kinetic parameters were then analyzed as a function of storage temperature, using the square root model, polynomial equation, and linear equation. A dynamic model was also developed for varying temperature. The model performance was evaluated against observed data, and the root mean square error (RMSE) was calculated. At 4°C, E. coli cell growth was not observed on any cheese. However, E. coli growth was observed at 10°C to 30°C with a μ max of 0.01 to 1.03 log CFU/g/h, depending on the cheese. The μ max values increased as temperature increased, while LPD values decreased, and μ max and LPD values were different among the four types of cheese. The developed models showed adequate performance (RMSE = 0.176-0.337), indicating that these models should be useful for describing the growth kinetics of E. coli on various cheeses.

Toys and toilets: cross-sectional study using children's toys to evaluate environmental faecal contamination in rural Bangladeshi households with different sanitation facilities and practices.

PubMed

Vujcic, Jelena; Ram, Pavani K; Hussain, Faruqe; Unicomb, Leanne; Gope, Partha Sarathi; Abedin, Jaynal; Mahmud, Zahid Hayat; Islam, M Sirajul; Luby, Stephen P

2014-05-01

To evaluate household faecal contamination using children's toys among 100 rural Bangladeshi households categorised as 'cleaner' (toilet that reliably separates faeces from the environment and no human faeces in/around living space) or 'less clean' (no toilet or toilet that does not reliably separate faeces from the environment and human faeces in/around living space). We distributed toy balls to each household and rinsed each study toy and a toy already owned by the household in 200 ml of Ringer's solution. We enumerated faecal coliforms and faecal streptococci from each rinse using membrane filtration methods. Study toys from 39 cleaner households had lower mean faecal coliform contamination than toys from 61 less clean households (2.4 log10 colony-forming units (CFU)/200 ml vs. 3.2 log10 CFU/200 ml, P = 0.03). However, wealth measures explained a portion of this relationship. Repeat measures were moderately variable [coefficient of variation (CV) = 6.5 between two toys in the household at the same time, CV = 37.6 between toys in the household at two different times 3-4 days apart]. Too few households owned a non-porous toy to compare groups without risk of bias. In rural Bangladesh, improved sanitation facilities and practices were associated with less environmental contamination. Whether this association is independent of household wealth and whether the difference in contamination improves child health merit further study. The variation found was typical for measures of environmental contamination, and requires large sample sizes to ascertain differences between groups with statistical significance. © 2014 John Wiley & Sons Ltd.

Effects of anticoagulant, processing delay, and assay method (branched DNA versus reverse transcriptase PCR) on measurement of human immunodeficiency virus type 1 RNA levels in plasma.

PubMed

Kirstein, L M; Mellors, J W; Rinaldo, C R; Margolick, J B; Giorgi, J V; Phair, J P; Dietz, E; Gupta, P; Sherlock, C H; Hogg, R; Montaner, J S; Muñoz, A

1999-08-01

We conducted two studies to determine the potential influence of delays in blood processing, type of anticoagulant, and assay method on human immunodeficiency virus type 1 (HIV-1) RNA levels in plasma. The first was an experimental study in which heparin- and EDTA-anticoagulated blood samples were collected from 101 HIV-positive individuals and processed to plasma after delays of 2, 6, and 18 h. HIV-1 RNA levels in each sample were then measured by both branched-DNA (bDNA) and reverse transcriptase PCR (RT-PCR) assays. Compared to samples processed within 2 h, the loss (decay) of HIV-1 RNA in heparinized blood was significant (P < 0.05) but small after 6 h (bDNA assay, -0.12 log(10) copies/ml; RT-PCR, -0.05 log(10) copies/ml) and after 18 h (bDNA assay, -0.27 log(10) copies/ml; RT-PCR, -0.15 log(10) copies/ml). Decay in EDTA-anticoagulated blood was not significant after 6 h (bDNA assay, -0.002 log(10) copies/ml; RT-PCR, -0.02 log(10) copies/ml), but it was after 18 h (bDNA assay, -0.09 log(10) copies/ml; RT-PCR, -0.09 log(10) copies/ml). Only 4% of samples processed after 6 h lost more than 50% (>/=0.3 log(10) copies/ml) of the HIV-1 RNA, regardless of the anticoagulant or the assay that was used. The second study compared HIV-1 RNA levels in samples from the Multicenter AIDS Cohort Study (MACS; samples were collected in heparin-containing tubes in 1985, had a 6-h average processing delay, and were assayed by bDNA assay) and the British Columbia Drug Treatment Program (BCDTP) (collected in EDTA- or acid citrate dextrose-containing tubes in 1996 and 1997, had a 2-h maximum processing delay, and were assayed by RT-PCR). HIV-1 RNA levels in samples from the two cohorts were not significantly different after adjusting for CD4(+)-cell count and converting bDNA assay values to those corresponding to the RT-PCR results. In summary, the decay of HIV-1 RNA measured in heparinized blood after 6 h was small (-0.05 to -0.12 log(10) copies/ml), and the minor impact of this decay on HIV-1 RNA concentrations in archived plasma samples of the MACS was confirmed by the similarity of CD4(+)-cell counts and assay-adjusted HIV-1 RNA concentrations in the MACS and BCDTP.

Lethal photosensitization of wound-associated microbes using indocyanine green and near-infrared light.

PubMed

Omar, Ghada S; Wilson, Michael; Nair, Sean P

2008-07-01

The increase in resistance to antibiotics among disease-causing bacteria necessitates the development of alternative antimicrobial approaches such as the use of light-activated antimicrobial agents (LAAAs). Light of an appropriate wavelength activates the LAAA to produce cytotoxic species which can then cause bacterial cell death via loss of membrane integrity, lipid peroxidation, the inactivation of essential enzymes, and/or exertion of mutagenic effects due to DNA modification. In this study, the effect of the LAAA indocyanine green excited with high or low intensity light (808 nm) from a near-infrared laser (NIR) on the viability of Staphylococcus aureus, Streptococcus pyogenes and Pseudomonas aeruginosa was investigated. All species were susceptible to killing by the LAAA, the bactericidal effect being dependent on both the concentration of indocyanine green and the light dose. Indocyanine green photosensitization using both high (1.37 W cm(-2)) and low (0.048 W cm(-2)) intensity NIR laser light was able to achieve reductions of 5.6 log10 (>99.99%) and 6.8 log10 (>99.99%) in the viable counts of Staph. aureus and Strep. pyogenes (using starting concentrations of 106-107 CFU ml(-1)). Kills of 99.99% were obtained for P. aeruginosa (initial concentration 108-109 CFU ml(-1)) photosensitized by the high intensity light (1.37 W cm(-2)); while a kill of 80% was achieved using low intensity irradiation (0.07 W cm(-2)). The effects of L-tryptophan (a singlet oxygen scavenger) and deuterium oxide (as an enhancer of the life span of singlet oxygen) on the survival of Staph. aureus was also studied. L-tryptophan reduced the proportion of Staph. aureus killed; whereas deuterium oxide increased the proportion killed suggesting that singlet oxygen was involved in the killing of the bacteria. These findings imply that indocyanine green in combination with light from a near-infrared laser may be an effective means of eradicating bacteria from wounds and burns.

Standardization of the CFU-GM assay: Advantages of plating a fixed number of CD34+ cells in collagen gels.

PubMed

Dobo, Irène; Pineau, Danielle; Robillard, Nelly; Geneviève, Frank; Piard, Nicole; Zandecki, Marc; Hermouet, Sylvie

2003-10-01

We investigated whether plating a stable amount of CD34(+) cells improves the CFU-GM assay. Data of CFU-GM assays performed with leukaphereses products in two transplant centers using a commercial collagen-based medium and unified CFU-GM scoring criteria were pooled and analyzed according to the numbers of CD34(+) cells plated. A first series of 113 CFU-GM assays was performed with a fixed number of mononuclear cells (i.e., a variable number of CD34(+) cells). In these cultures the CFU-GM/CD34 ratio varied according to the number of CD34(+) cells plated: median CFUGM/CD34 ratios were 1/6.2 to 1/6.6 for grafts containing <2% CD34(+) cells, vs. 1/10.2 for grafts containing > or =2% CD34(+) cells. The median CFU-GM/CD34 ratio also varied depending on pathology: 1/9.3 for multiple myeloma (MM), 1/6.8 for Hodgkin's disease (HD), 1/6.5 for non-Hodgkin lymphoma (NHL), and 1/4.5 for solid tumors (ST). A second series of 95 CFU-GM assays was performed with a fixed number of CD34(+) cells (220/ml). The range of median CFU-GM/CD34 ratios was narrowed to 1/7.0 to 1/5.2, and coefficients of variation for CFU-GM counts decreased by half to 38.1% (NHL), 36.1% (MM), 49.9% (HD), and 22.4% (ST). In addition, CFU-GM scoring was facilitated as the percentages of cultures with >50 CFU/GM/ml decreased from 6.7% to 43.8% when a variable number of CD34(+) cells was plated, to 4.5% to 16.7% when 220 CD34(+) cells/ml were plated. Hence, plating a fixed number of CD34(+) cells in collagen gels improves the CFU-GM assay by eliminating cell number-related variability and reducing pathology-related variability in colony growth.

Pilot-scale production and liquid formulation of Rhodotorula minuta, a potential biocontrol agent of mango anthracnose.

PubMed

Patiño-Vera, M; Jiménez, B; Balderas, K; Ortiz, M; Allende, R; Carrillo, A; Galindo, E

2005-01-01

To develop a pilot-plant fermentation process for the production of the yeast Rhodotorula minuta, to be used as a biocontrol agent of mango anthracnose, using a low-cost culture medium. To develop a stable liquid formulation that preserve high viability of the yeast stored at 4 degrees C. Keeping constant the volumetric power input, a fermentation process was scaled-up from shake flasks to a 100 l bioreactor. Preharvest applications of the yeast resulted in postharvest anthracnose severity equal or lower than that observed with a chemical fungicide. Glycerol was added to the formulation as water activity reducer and xanthan gum as a viscosity-enhancing agent. Yeast initial concentration of 10(10) CFU ml(-1) resulted in 4-5 orders of magnitude decrease after 1 month of storage at 4 degrees C, whereas when it was formulated at 10(9) CFU ml(-1), the decrease was of two orders of magnitude in 6 months. The fermentation process was successfully scaled-up using a low-cost culture medium. Postharvest anthracnose severity could be considerably reduced using this yeast. Formulating the yeast at 10(9) CFU ml(-1) and adding glycerol (20%) and xanthan (5 g l(-1)) avoided both contamination and yeast sedimentation and it was able to preserve up to 10(7) CFU ml(-1) after 6 months at 4 degrees C. The yeast R. minuta is reported as a novel antagonistic micro-organism against the pathogen Colletotrichum gloeosporioides. Pilot plant production of this yeast allowed us to conduct field tests in commercial orchards during three harvest seasons. Yeast suspensions applied to mango trees reduced the fruit anthracnose severity in levels similar or better than chemical fungicides.

Fate of Yersinia enterocolitica during manufacture, ripening and storage of Lighvan cheese.

PubMed

Hanifian, Shahram; Khani, Sajjad

2012-05-15

This study aimed to evaluate the behavior of virulent Yersinia enterocolitica (YE) during the manufacture, ripening and storage of Lighvan cheese with particular reference to strains of YE, initial inoculation level, and storage time. Three strains of YE with low (1 log cfu/ml) and high (3 log cfu/ml) inoculation levels were inoculated to raw whole ewe's milk which was then used for manufacturing of Lighvan cheese. Throughout the manufacturing, ripening and storage periods the number of YE was counted on selective media. Enumerated colonies were then confirmed by duplex PCR using ail and virF genes. Moreover, some microbial and physiochemical characteristics of the cheese samples were examined. According to the results, initial inoculation level and storage time had statistically significant (P<0.01) effects on persistency of YE, while strain type exhibited no statistically significant (P>0.01) impact on survival of the pathogen. Results showed a rapid increase in the number of YE during manufacturing, however, in the ripening and storage periods the number of YE was decreased and eventually it was eliminated in all cheese batches after 4 months of storage. Copyright © 2012 Elsevier B.V. All rights reserved.

Inactivation of Salmonella enterica and Listeria monocytogenes in cantaloupe puree by high hydrostatic pressure with/without added ascorbic acid.

PubMed

Mukhopadhyay, Sudarsan; Sokorai, Kimberly; Ukuku, Dike; Fan, Xuetong; Juneja, Vijay; Sites, Joseph; Cassidy, Jennifer

2016-10-17

The objective of this research was to evaluate and develop a method for inactivation of Salmonella enterica and Listeria monocytogenes in cantaloupe puree (CP) by high hydrostatic pressure (HHP). Cantaloupe being the most netted varieties of melons presents a greater risk of pathogen transmission. Freshly prepared CP with or without 0.1% ascorbic acid (AA) was inoculated with a bacterial cocktail composed of a three serotype mixture of S. enterica (S. Poona, S. Newport H1275 and S. Stanley H0558) and a mixture of three strains of L. monocytogenes (Scott A, 43256 and 51742) to a population of ca. 10(8)CFU/g. Double sealed and double bagged inoculated CP (ca. 5g) were pressure treated at 300, 400 and 500MPa at 8°C and 15°C for 5min. Data indicated increased inactivation of both Salmonella and Listeria spp. with higher pressure. Log reduction for CP at 300MPa, 8°C for 5min was 2.4±0.2 and 1.6±0.5logCFU/g for Salmonella and Listeria, respectively. Survivability of the pathogens was significantly compromised at 400MPa and 8°C, inactivating 4.5±0.3logCFU/g of Salmonella and 3.0±0.4logCFU/g of Listeria spp. Complete inactivation of the pathogens in the puree (log reduction >6.7logCFU/g), with or without AA, was achieved when the pressure was further increased to 500MPa, except that for Listeria containing no AA at 8°C. Listeria presented higher resistance to pressure treatment compared to Salmonella spp. Initial temperatures (8 and 15°C) had no significant influence on Salmonella log reductions. Log reduction of pathogens increased but not significantly with increase of temperature. AA did not show any significant antimicrobial activity. Viable counts were about 0.2-0.4logCFU/g less in presence of 0.1% AA. These data validate that HHP can be used as an effective method for decontamination of cantaloupe puree. Published by Elsevier B.V.

Preliminary Studies on Membrane Filtration for the Production of Potable Water: A Case of Tshaanda Rural Village in South Africa

PubMed Central

Molelekwa, Gomotsegang F.; Mukhola, Murembiwa S.; Van der Bruggen, Bart; Luis, Patricia

2014-01-01

Ultrafiltration (UF) systems have been used globally for treating water from resources including rivers, reservoirs, and lakes for the production of potable water in the past decade. UF membranes with a pore size of between 0.1 and 0.01 micrometres provide an effective barrier for bacteria, viruses, suspended particles, and colloids. The use of UF membrane technology in treating groundwater for the supply of potable water in the impoverished and rural village, Tshaanda (i.e., the study area) is demonstrated. The technical and administrative processes that are critical for the successful installation of the pilot plant were developed. Given the rural nature of Tshaanda, the cultural and traditional protocols were observed. Preliminary results of the water quality of untreated water and the permeate are presented. Escherichia coli in the untreated water during the dry season (i.e., June and July) was 2 cfu/100 ml and was <1 cfu/100 ml (undetected) following UF, which complied with the WHO and South African National Standards and Guidelines of <1 cfu/100 ml. During the wet/rainy season (February) total coliform was unacceptably high (>2419.2 cfu/100 ml) before UF. Following UF, it dramatically reduced to acceptable level (7 cfu/100 ml) which is within the WHO recommended level of <10 cfu/100 ml. Additionally, during the wet/rainy season E. coli and enterococci were unacceptably high (40.4 cfu/100 ml and 73.3 cfu/100 ml, respectively) before UF but were completely removed following UF, which are within the WHO and SANS recommended limit. The values for electrical conductivity (EC) and turbidity were constantly within the WHO recommended limits of 300 µS/cm corrected at 25°C and <5 NTU, respectively, before and after UF, during dry season and wet season. This suggests that there is no need for pre-treatment of the water for suspended particles and colloids. Considering these data, it can be concluded that the water is suitable for human consumption, following UF. PMID:25144640

Preliminary studies on membrane filtration for the production of potable water: a case of Tshaanda rural village in South Africa.

PubMed

Molelekwa, Gomotsegang F; Mukhola, Murembiwa S; Van der Bruggen, Bart; Luis, Patricia

2014-01-01

Ultrafiltration (UF) systems have been used globally for treating water from resources including rivers, reservoirs, and lakes for the production of potable water in the past decade. UF membranes with a pore size of between 0.1 and 0.01 micrometres provide an effective barrier for bacteria, viruses, suspended particles, and colloids. The use of UF membrane technology in treating groundwater for the supply of potable water in the impoverished and rural village, Tshaanda (i.e., the study area) is demonstrated. The technical and administrative processes that are critical for the successful installation of the pilot plant were developed. Given the rural nature of Tshaanda, the cultural and traditional protocols were observed. Preliminary results of the water quality of untreated water and the permeate are presented. Escherichia coli in the untreated water during the dry season (i.e., June and July) was 2 cfu/100 ml and was <1 cfu/100 ml (undetected) following UF, which complied with the WHO and South African National Standards and Guidelines of <1 cfu/100 ml. During the wet/rainy season (February) total coliform was unacceptably high (>2419.2 cfu/100 ml) before UF. Following UF, it dramatically reduced to acceptable level (7 cfu/100 ml) which is within the WHO recommended level of <10 cfu/100 ml. Additionally, during the wet/rainy season E. coli and enterococci were unacceptably high (40.4 cfu/100 ml and 73.3 cfu/100 ml, respectively) before UF but were completely removed following UF, which are within the WHO and SANS recommended limit. The values for electrical conductivity (EC) and turbidity were constantly within the WHO recommended limits of 300 µS/cm corrected at 25°C and <5 NTU, respectively, before and after UF, during dry season and wet season. This suggests that there is no need for pre-treatment of the water for suspended particles and colloids. Considering these data, it can be concluded that the water is suitable for human consumption, following UF.

Exposure to biohazards in wood dust: bacteria, fungi, endotoxins, and (1-->3)-beta-D-glucans.

PubMed

Alwis, K U; Mandryk, J; Hocking, A D

1999-09-01

Personal exposure to fungi, bacteria, endotoxin, and (1-->3)-beta-D-glucan was determined at different woodworking sites--logging sites, sawmills, woodchipping sites, and joineries. Exposure levels to fungi at logging sites and sawmills were in the range of 10(3)-10(4) cfu/m3, at the woodchipping mill, 10(3)-10(5) cfu/m3, and at joineries, 10(2)-10(4) cfu/m3. Although mean endotoxin levels were lower than the suggested threshold value of 20 ng/m3, some personal exposures at sawmills and a joinery exceeded the standard. The geometric mean personal (1-->3)-beta-D-glucan exposure level at the woodchipping mill was 2.32 ng/m3, at sawmills, 1.37 ng/m3, at logging sites, 2.02 ng/m3, and at joineries, 0.43 ng/m3. Highly significant associations were found between mean personal inhalable endotoxin exposures and Gram-negative bacteria levels (p < 0.0001), and mean personal inhalable (1-->3)-beta-D-glucan exposures and fungi levels (p = 0.0003). The prevalence of cough, phlegm, chronic bronchitis, nasal symptoms, frequent headaches, and eye and throat irritations was significantly higher among woodworkers than controls. Dose-response relationships were found between personal exposures and work-related symptoms among joinery workers and sawmill and chip mill workers.

Antimicrobial activity of Hibiscus sabdariffa aqueous extracts against Escherichia coli O157:H7 and Staphylococcus aureus in a microbiological medium and milk of various fat concentrations.

PubMed

Higginbotham, Kristen L; Burris, Kellie P; Zivanovic, Svetlana; Davidson, P Michael; Stewart, C Neal

2014-02-01

Hibiscus sabdariffa L. calyces are widely used in the preparation of beverages. The calyces contain compounds that exhibit antimicrobial activity, yet little research has been conducted on their possible use in food systems as antimicrobials. Aqueous extracts prepared from the brand "Mi Costenita" were sterilized by membrane filtration (0.22-μm pore size) or autoclaving (121 °C, 30 min) and tested for antimicrobial activity against the foodborne pathogens Escherichia coli O157:H7 strains ATCC 43894 and Cider and Staphylococcus aureus strains SA113 and ATCC 27708 in a microbiological medium and ultrahigh-temperature-processed milk with various fat percentages. Extracts heated by autoclaving exhibited greater activity than did filtered extracts in a microbiological medium. Against E. coli, results of 20 mg/ml filtered extract were not different from those of the control, whereas autoclaved extracts reduced viable cells ca. 3 to 4 log CFU/ml. At 60 mg/ml, both extracts inactivated cells after 24 h. There were reduced populations of both strains of S. aureus (ca. 2.7 and 3 log CFU/ml, respectively) after 24 h of incubation in 40 mg/ml filtered extracts. When grown in autoclaved extracts at 40 mg/ml, both strains of S. aureus were inactivated after 9 h. Autoclaved extracts had decreased anthocyanin content (2.63 mg/liter) compared with filtered extracts (14.27 mg/liter), whereas the phenolic content (48.7 and 53.8 mg/g) remained similar for both treatments. Autoclaved extracts were then tested for activity in milk at various fat concentrations (skim [<0.5%], 1%, 2%, and whole [>3.25%]) against a 1:1 mixture of the two strains of E. coli O157:H7 and a 1:1 mixture of the two strains of S. aureus. Extracts at 40 mg/ml inactivated S. aureus after 168 h in skim and whole milk, and E. coli was inactivated after 96 h in 60 mg/ml extract in all fat levels. These findings show the potential use of Hibiscus extracts to prevent the growth of pathogens in foods and beverages.

Effect of phage and host concentration on the inactivation of Escherichia coli O157:H7 on cooked and raw beef.

PubMed

Hudson, J A; Billington, C; Wilson, T; On, S L W

2015-03-01

A previously described phage infecting Escherichia coli O157:H7 was added to raw and cooked beef pieces at concentrations ranging from 10(1)-10(8) plaque forming units/cm(2) to either low (<100 CFU/cm(2)) or high (10(4) CFU/cm(2)) concentrations of host bacterial cells. Incubation for up to 24 h was performed at 5 ℃ and 24 ℃ to simulate refrigerated and room temperature storage/temperature abuse. Surviving bacteria were enumerated during the incubation period, with phages being counted at the first and last sampling times. Significant reductions of E. coli O157:H7 of the order of >4 log10 CFU/cm(2) at both temperatures could be achieved compared to phage-free controls. There was a trend for greater inactivation to occur with increasing phage concentration. While re-growth of surviving cells occurred in nearly all samples incubated for 24 h at 24 ℃, these conditions are not typical of those experienced by perishable foods. It was concluded that phages can be used to reduce the concentration of a bacterial pathogen on meat, but the concentration of phages needs to be high (>4-5 log10 plaque forming units/cm(2)) for reductions to occur. A concentration of the order 8 log10 plaque forming units/cm(2) was needed to achieve a 4 log10 CFU/cm(2) reduction. © The Author(s) 2013 Reprints and permissions: sagepub.co.uk/journalsPermissions.nav.

Leakage of Intracellular UV Materials of High Hydrostatic Pressure-Injured Escherichia Coli O157:H7 Strains in Tomato Juice

USDA-ARS?s Scientific Manuscript database

The effect of high hydrostatic pressure (HHP) treatment on inactivation, injury and recovery of Salmonella Enteritidis and Escherichia coli O157:H7 cocktail inoculated in tomato juice (pH 4.1) and phosphate buffer saline (PBS. pH 7.2) at 8.0 log CFU/ml and treated at 350, 400, 450 MPa for 20 min at ...

Kefir alleviates obesity and hepatic steatosis in high-fat diet-fed mice by modulation of gut microbiota and mycobiota: targeted and untargeted community analysis with correlation of biomarkers.

PubMed

Kim, Dong-Hyeon; Kim, Hyunsook; Jeong, Dana; Kang, Il-Byeong; Chon, Jung-Whan; Kim, Hong-Seok; Song, Kwang-Young; Seo, Kun-Ho

2017-06-01

Kefir is a probiotic beverage containing over 50 species of lactic acid bacteria and yeast. In this study, the anti-obesity and anti-non-alcoholic fatty liver disease (NAFLD) effects of kefir were comprehensively addressed along with targeted and untargeted community analysis of the fecal microbiota in a high-fat diet (HFD)-induced obese mouse model. HFD-fed C57BL/6 mice were orally administrated either kefir or milk (control) once a day for 12 weeks, and body and organ weight, fecal microbiota and mycobiota, histopathology, blood cholesterol and cytokines and gene expressions were analyzed. Compared to the control, mice in the kefir group exhibited a significantly lower body weight (34.18 g vs. 40.24 g; p=0.00004) and histopathological liver lesion score (1.13 vs. 3.25; p=0.002). Remarkably, the kefir-fed mice also harbored more Lactobacillus/Lactococcus (7.01 vs. 6.32 log CFU/g), total yeast (6.07 vs. 5.01 log CFU/g) and Candida (5.56 vs. 3.88 log CFU/g). Kefir administration also up-regulated genes related to fatty acid oxidation, PPARα and AOX, in both the liver and adipose tissue (PPARα, 2.95- and 2.15-fold; AOX, 1.89- and 1.9-fold, respectively). The plasma concentration of IL-6, a proinflammatory marker, was significantly reduced following kefir consumption (50.39 pg/ml vs. 111.78 pg/ml; p=0.03). Strikingly, the populations of Lactobacillus/Lactococcus, total yeast and Candida were strongly correlated with PPARα gene expression in adipose and hepatic tissue (r=0.599, 0.580 and 0.562, respectively). These data suggest that kefir consumption modulates gut microbiota and mycobiota in HFD-fed mice, which prevents obesity and NAFLD via promoting fatty acid oxidation. Copyright © 2017 Elsevier Inc. All rights reserved.

Acid and alcohol tolerance of Escherichia coli O157:H7 in pulque, a typical Mexican beverage.

PubMed

Gómez-Aldapa, Carlos A; Díaz-Cruz, Claudio A; Villarruel-López, Angelica; Del Refugio Torres-Vitela, M; Rangel-Vargas, Esmeralda; Castro-Rosas, Javier

2012-03-01

Pulque is a traditional Mexican fermented alcoholic beverage produced from the nectar of maguey agave plants. No data exist on the behavior of Escherichia coli O157:H7 in agave nectar and pulque. An initial trial was done of the behavior of E. coli O157:H7 during fermentation of nectar from a single producer, a nectar mixture from different producers and "seed" pulque. A second trial simulating artisanal pulque production was done by contaminating fresh nectar with a cocktail of three E. coli O157:H7 strains, storing at 16 ° and 22 °C for 14 h, adding seed pulque and fermenting until pulque was formed. A third trial used pulque from the second trial stored at 22 °C as seed to ferment fresh nectar at 22 °C for 48 h (fermentation cycle). This procedure was repeated for an additional two fermentation cycles. During incubation at 16 ° or 22 °C in the first trial, the E. coli O157:H7 strains multiplied in both the single producer nectar and nectar mixture, reaching maximum concentration at 12h. E. coli O157:H7 cell concentration then decreased slowly, although it survived at least 72 h in both fermented nectars. E. coli O157:H7 did not multiply in the seed pulque but did survive at least 72 h. In the second trial, the numbers of E. coli O157:H7 increased approximately 1.5 log CFU/ml at 22 °C and 1.2 log CFU/ml at 16 °C after 14 h. After seed pulque was added, E. coli O157:H7 concentration decreased to approximately 2 log CFU/ml, and then remained constant until pulque was produced. In the third trial, the E. coli O157:H7 cells multiplied and survived during at least three nectar fermentation cycles. The results suggest that E. coli O157:H7 can develop acid and alcohol tolerance in pulque, and constitutes a public health risk for pulque consumers. Copyright © 2011 Elsevier B.V. All rights reserved.

Activity of Tedizolid in Methicillin-Resistant Staphylococcus epidermidis Experimental Foreign Body-Associated Osteomyelitis.

PubMed

Park, Kyung-Hwa; Greenwood-Quaintance, Kerryl E; Schuetz, Audrey N; Mandrekar, Jayawant N; Patel, Robin

2017-02-01

We developed a rat model of methicillin-resistant Staphylococcus epidermidis (MRSE) foreign body-associated osteomyelitis and used it to compare tedizolid alone and in combination with rifampin against rifampin alone, vancomycin plus rifampin, and vancomycin alone. A clinical strain of MRSE was inoculated into the proximal tibia, and a stainless steel wire with a precolonized MRSE biofilm was implanted. Following a 1-week infection period, 92 rats received either no treatment (n = 17) or 14 days of intraperitoneal tedizolid (n = 15), tedizolid plus rifampin (n = 15), rifampin (n = 15), vancomycin plus rifampin (n = 15), or vancomycin (n = 15). Quantitative bone and wire cultures were performed after treatment completion and also 1 week after infection in a separate group of five rats. The median quantity of staphylococci in bone after the 1-week infection period was 4.89 log 10 CFU/g bone (interquartile range, 3.83 to 5.33 log 10 CFU/g bone); staphylococci were recovered from all associated wires. A median quantity of staphylococci of 3.70 log 10 CFU/g bone was detected in bones of untreated control rats after 3 weeks. Quantities of staphylococci in bones of all treatment groups except the group receiving vancomycin alone (2.78 log 10 CFU/g) were significantly lower than those for untreated controls, with no staphylococci being detected in the groups receiving rifampin monotherapy, tedizolid-plus-rifampin combination therapy, and vancomycin-plus-rifampin combination therapy. Quantities of staphylococci on wires from all treatment groups that included rifampin were significantly lower than those for untreated controls. No resistance to rifampin, tedizolid, or vancomycin was detected. Tedizolid combined with rifampin was active in a rat model of MRSE foreign body-associated osteomyelitis. Copyright © 2017 American Society for Microbiology.

Growth and persistence of pathogens on granular activated carbon filters.

PubMed Central

Camper, A K; LeChevallier, M W; Broadaway, S C; McFeters, G A

1985-01-01

Three enteric pathogens Yersinia enterocolitica O:8, Salmonella typhimurium, and enterotoxigenic Escherichia coli, were examined for their ability to colonize granular activated carbon (GAC) in pure cultures and in the presence of autochthonous river water organisms. All three organisms readily colonized sterile GAC and maintained populations of ca. 10(5) to 10(7) CFU g-1 for 14 days when suspended in sterile river water. Exposure of pathogen biofilms on GAC to unsterile river water resulted in a gradual decline in pathogens on the carbon (0.08 to 0.14 log day-1). When pathogens were introduced to sterile GAC in the presence of heterotrophic plate count organisms, they attached at levels similar to those in the pure cultures and then decreased (0.10 to 0.22 log day-1). When added with heterotrophic plate count bacteria to GAC supporting a mature biofilm of native river water bacteria, they attached at a lower level (1.0 X 10(4) to 4.6 X 10(4) CFU g-1) and decreased at a more rapid rate (0.11 to 0.70 log day-1). PMID:3911903

Growth of Legionella anisa in a model drinking water system to evaluate different shower outlets and the impact of cast iron rust.

PubMed

van der Lugt, Wilco; Euser, Sjoerd M; Bruin, Jacob P; Den Boer, Jeroen W; Walker, Jimmy T; Crespi, Sebastian

2017-11-01

Legionella continues to be a problem in water systems. This study investigated the influence of different shower mixer faucets, and the influence of the presence of cast iron rust from a drinking water system on the growth of Legionella. The research is conducted using a model of a household containing four drinking water systems. All four systems, which contained standard plumbing components including copper pipes and a water heater, were filled with unchlorinated drinking water. Furthermore, all systems had three different shower faucets: (A) a stainless-steel faucet, (B) a brass-ceramic faucet, and (C) a brass thermostatic faucet. System 1 was solely filled with drinking water. System 2 was filled with drinking water, and cast iron rust. System 3 was contaminated with Legionella, and system 4 was contaminated with a Legionella, and cast iron rust. During a period of 34 months, 450 cold water samples were taken from 15 sample points of the four drinking water systems, and tested for Legionella according to the Dutch Standard (NEN 6265). In system 4, with added cast iron rust, the stainless-steel mixer faucet (A) had the highest concentration of Legionella at >4.3log10CFU/l (>20,000CFU/l) and was positive in 46.4% of samples. In contrast, the stainless-steel mixer faucet (A) of system 3 without cast iron rust showed 14.3% positive samples with a maximum concentration of 3.9log10CFU/l (7600CFU/l) Legionella. Additionally, both contaminated systems (3 and 4), with the brass thermostatic faucet (C), tested positive for Legionella. System 3 in 85.7% of the samples, with a maximum concentration of 4.38log10CFU/l (24,200CFU/l), and system 4 in 64.3% of the samples with a maximum concentration of 4.13log10CFU/l (13.400CFU/l). These results suggest that both the type of faucet used in a drinking water system and the presence or absence of cast iron rust influence the growth of Legionella. Copyright © 2017 Elsevier GmbH. All rights reserved.

Contamination of hospital tap water: the survival and persistence of Pseudomonas aeruginosa on conventional and 'antimicrobial' outlet fittings.

PubMed

Hutchins, C F; Moore, G; Thompson, K-A; Webb, J; Walker, J T

2017-10-01

Pseudomonas aeruginosa infections have been linked to contaminated hospital taps, highlighting the potential for tap outlet fittings (OF) to harbour biofilm. P. aeruginosa may be transferred to OFs via contaminated cleaning cloths. Suggested interventions include flushing regimens and alternative OF designs. To investigate the transfer of P. aeruginosa from a contaminated cleaning cloth to conventional and 'antimicrobial/antibiofilm' OFs and to determine whether this contamination persists and/or leads to contamination of tap water. Microfibre cloths contaminated with P. aeruginosa (10 8  cfu/mL) were used to wipe four different types of OF [one of conventional design (OF-A) and three marketed as 'antimicrobial' and/or 'antibiofilm' (OF- B, -C and -D)]. OFs were inserted into an experimental water distribution system for up to 24 h. Survival was assessed by culture. Single and multiple water samples were collected and cultured for P. aeruginosa. The median number of P. aeruginosa transferred from cloth to OF was 5.7 × 10 5  cfu (OF-A), 1.9 × 10 6  cfu (OF-B), 1.4 × 10 5  cfu (OF-C) and 2.9 × 10 6  cfu (OF-D). Numbers declined on all OFs during the 24 h period with log reductions ranging from 3.5 (OF-C) to 5.2 (OF-B; P > 0.05). All water samples delivered immediately after OF contamination contained P. aeruginosa at ≥10 cfu per 100 mL. Contamination of water delivered from OF-A persisted despite continued flushing. Water delivered from OF-B did not contain P. aeruginosa beyond the first flush. Contaminated cleaning cloths may transfer P. aeruginosa to OFs, leading to contamination of tap water. Although not removing the potential for contamination, 'antimicrobial/antibiofilm' OFs may prevent P. aeruginosa from continually contaminating water delivered from the outlet. Copyright © 2017 The Healthcare Infection Society. All rights reserved.

Microbiological Quality and Incidence of Salmonella on Cherry Tomatoes at Retail in Querétaro, México.

PubMed

Leal-Cervantes, Marla G; Arvizu-Medrano, Sofía M; Martínez-Peniche, Ramón; Martínez-Gonzáles, Nanci E; Hernández-Iturriaga, Montserrat

2018-04-01

Multiple outbreaks related to Salmonella in tomatoes require an evaluation of the risk associated with cherry tomatoes due to the increase in its production, consumption, and marketing in Mexico's central region. The purpose of this study was to determine the microbial quality of cherry tomatoes obtained from two retail sale points (supermarkets and local markets). Cherry tomato samples (333) were collected from four supermarkets and from four local markets, and the contents of aerobic plate count, molds and yeasts, total coliforms, and Escherichia coli were quantified; the presence of Salmonella was simultaneously determined. The median values of the microbial populations were obtained, and the data were analyzed per the sampling site by using the Wilcoxon and Kruskal-Wallis tests. The median of aerobic plate count content in tomatoes obtained from supermarkets ranged between 2.2 and 4.4 log CFU/g, and in markets from 2.9 to 4.8 log CFU/g. For molds and yeasts, the tomatoes from supermarkets (2.0 to 4.1 log CFU/g) and markets (1.5 to 4.5 log CFU/g) showed similar contents. Regardless of the sampling site, the values of total coliforms were very low, ranging from 1.0 to 1.8 log CFU/g. E. coli was detected in 5.4 and 20.1% of samples from supermarkets and markets, respectively; in both sites, the content was low (0.3 to 5.8 most probable number per g). The incidence of Salmonella was 14.1% in supermarkets and 7.8% in local markets. The results obtained from this investigation highlight the elevated risk for consumer health associated with the ingestion of cherry tomatoes.

Application of high pressure processing to reduce verotoxigenic E. coli in two types of dry-fermented sausage.

PubMed

Omer, M K; Alvseike, O; Holck, A; Axelsson, L; Prieto, M; Skjerve, E; Heir, E

2010-12-01

The effect of high pressure processing (HPP) on the survival of verotoxigenic Escherichia coli (VTEC) in two types of Norwegian type dry-fermented sausages was studied. Two different types of recipes for each sausage type were produced. The sausage batter was inoculated with 6.8 log(10) CFU/g of VTEC O103:H25. After fermentation, drying and maturation, slices of finished sausages were vacuum packed and subjected to two treatment regimes of HPP. One group was treated at 600 MPa for 10 min and another at three cycles of 600 MPa for 200 s per cycle. A generalized linear model split by recipe type showed that these two HPP treatments on standard recipe sausages reduced E. coli by 2.9 log(10) CFU/g and 3.3 log(10) CFU/g, respectively. In the recipe with higher levels of dextrose, sodium chloride and sodium nitrite E. coli reduction was 2.7 log(10) CFU/g in both treatments. The data show that HPP has a potential to make the sausages safer and also that the effect depends somewhat on recipe. Copyright © 2010 The American Meat Science Association. Published by Elsevier Ltd. All rights reserved.

Behavior of radio frequency electric fields injured Escherichia coli in nutrient and non nutrient media during storage

USDA-ARS?s Scientific Manuscript database

Information on conditions required for resuscitation of Radio Frequency Electric Fields (RFEF) damaged E. coli cells is limited. Apple juice inoculated with Escherichia coli K-12 at 7.8 log CFU/ml was treated with RFEF at 20 kHz, 15 kV/cm for 170 micro second at 55C with a flow rate of 540 ml/min. A...

Lack of antimicrobial effect on periodontopathic bacteria by ultrasonic and sonic scalers in vitro.

PubMed

Schenk, G; Flemmig, T F; Lob, S; Ruckdeschel, G; Hickel, R

2000-02-01

The purpose of this study was to assess the antimicrobial effects of a sonic and ultrasonic scaler generally used for subgingival scaling on gram-negative and gram-positive periodontopathic bacteria. Actinobacillus actinomycetemcomitans, Porphyromonas gingivalis, Campylobacter rectus, or Peptostreptococcus micros were suspended in Schaedler's broth medium and treated by a sonic or a magnetostrictive ultrasonic scaler for 30 s and 150 s in vitro. Bacterial suspensions treated by an ultrasonic cell disruptor served as a positive control and untreated bacterial suspensions served as a negative control. Following sonication, samples were serially diluted, streaked on blood agar plates and incubated for 2-5 days at 37 degrees C. Treatment by the sonic or ultrasonic scaler for up to 150 s did not reduce the viability of any of the tested periodontal pathogens. Compared to untreated controls, the viability of A. actinomycetemcomitans and P. gingivalis was significantly (p<0.05) reduced only following ultrasonication with the cell disruptor after 30 s (0.72 and 0.54 log CFU/ml, respectively) and of A. actinomycetemcomitans, P. gingivalis, C. rectus, and P. micros after 150 s (1.98, 1.34, 1.95 and 1.98 log CFU/ml, respectively). The data of the study may indicate that the assessed sonic and ultrasonic scaler used for subgingival debridement do not result in killing of the tested periodontal pathogens.

Benzimidazole-Based Antibacterial Agents Against F. tularensis

PubMed Central

Kumar, Kunal; Awasthi, Divya; Lee, Seung-Yub; Cummings, Jason E.; Knudson, Susan E.; Slayden, Richard A.; Ojima, Iwao

2013-01-01

Francisella tularensis is a highly virulent pathogenic bacterium. In order to identify novel potential antibacterial agents against F. tularensis, libraries of trisubstituted benzimidazoles were screened against F. tularensis LVS strain. In a preliminary screening assay, remarkably, 23 of 2,5,6- and 2,5,7-trisubstituted benzimidazoles showed excellent activity exhibiting greater than 90 % growth inhibition at 1 µg/mL. Among those hits, 21 compounds showed MIC90 values in the range of 0.35–48.6 µg/mL after accurate MIC determination. In ex-vivo efficacy assays, four of these compounds exhibited 2–3 Log reduction in colony forming units (CFU) per mL at concentrations of 10 and 50 µg/mL. PMID:23623254

Assessment of microbiological quality of sachet-packaged drinking water in Western Nigeria and its public health significance.

PubMed

Olaoye, O A; Onilude, A A

2009-11-01

To assess the microbiological quality of sachet-packaged drinking water in Western Nigeria and its impact on public health. Cross-sectional microbiological testing. Ninety-two sachet-packaged water samples were analysed for microbiological and metal qualities. Total bacterial and coliform counts were determined, and the presence of Escherichia coli, an important water quality indicator, was tested. The level of conformity of the water processors with the guidelines of Nigeria's quality regulatory agency was also determined. Varying levels of microbial contamination were recorded in samples from the different sampling locations. The total bacteria count ranged between 2.86 and 3.45log colony-forming units (cfu)/ml. The highest coliform count recorded was 1.62log cfu/ml. Faecal coliform E. coli was detected in one sample from Oke-Iho and one sample from Okaka, representing 2.2% of total samples. Lead and manganese were not found in any of the samples. However, iron was detected and the highest iron concentration (0.10mg/l) was detected in samples from Ikorodu. The bacteria that were identified from the water samples included E. coli, Pseudomonas aeruginosa, Enterobacter aerogenes, Klebsiella sp., Proteus vulgaris, Alcaligenes faecalis, Bacillus cereus, Staphylococcus aureus, Streptococcus lactis, Aeromonas sp. and Micrococcus luteum. Many of the water processors did not comply with the guidelines of the quality regulatory agency. Some of the sachet-packaged samples of drinking water were of poor quality. The results indicate a need for Nigeria's quality regulatory agency to take appropriate measures in safeguarding public health.

Efficacy of humanlike Augmentin SR (2000/125 mg) twice daily treatment on Haemophilus influenzae experimental pneumonia in rabbits.

PubMed

Croisier, Delphine; Benoit, Martha; Durand, David; Lequeu, Catherine; Piroth, Lionel; Portier, Henri; Chavanet, Pascal

2007-01-01

We investigated the efficacy of 2 formulations of Augmentin on experimental pneumonia due to Haemophilus influenzae (HI) in rabbits. Two strains were used (H128 and 401285) with amoxicillin/clavulanic acid MICs of 1/0.5 mg/l and 4/2 mg/l. Pneumonia was induced in immunocompetent rabbits by inoculation of 10 log(10) CFU HI. The treatments were infused by using computer controlled pumps in order to mimic the human pharmacokinetic (PK) profile of either conventional Augmentin treatment (875/125 mg twice daily) or the sustained release formulation (SR: 2000/125 mg twice daily). After 2 d of treatment, the bacterial concentrations in the lungs were similar for both strains and both treatments: isolate H128, conventional Augmentin reduced bacterial numbers to 3.8+/-2.1 log(10) CFU/g and Augmentin SR to 3.1+/-2.4 log(10) CFU/g; isolate 401285, conventional Augmentin to 3.5+/-2. Thus, both treatments demonstrated similar efficacy against H. influenzae pneumonia in this model, even when induced by a strain with an amoxicillin/clavulanic acid MIC of 4/2 mg/l. These results support current breakpoints for conventional Augmentin against H. influenzae and suggest that Augmentin SR is at least as effective against these isolates.

Bactericidal activity of lemon juice and lemon derivatives against Vibrio cholerae.

PubMed

de Castillo, M C; de Allori, C G; de Gutierrez, R C; de Saab, O A; de Fernandez, N P; de Ruiz, C S; Holgado, A P; de Nader, O M

2000-10-01

Food products can be possible vectors of the agent responsible for cholera epidemics, because some of these products allow Vibrio cholerae O1 to develop to concentrations above the dangerous level. This study deals with the behaviour of essential oils, natural and concentrated lemon juice and fresh and dehydrated lemon peel against V. cholerae O1 biotype Eltor serotype Inaba tox+. Our aim was to evaluate whether these products, used at different dilutions, exhibit bactericidal or bacteriostatic activity against the microorganism, when present at concentrations of 10(2), 10(4), 10(6) and 10(8) colony forming units (CFU) ml(-1), and after different exposure times. 10(8) CFU ml(-1) was considered an infectious dose. Concentrated lemon juice and essential oils inhibited V. cholerae completely at all studied dilutions and exposure times. Fresh lemon peel and dehydrated lemon peel partially inhibited growth of V. cholerae. Freshly squeezed lemon juice, diluted to 10(-2), showed complete inhibition of V. cholerae at a concentration of 10(8) CFU ml(-1) after 5 min of exposure time; a dilution of 2 x 10(-3) produced inhibition after 15 min and a dilution of 10(-3) after 30 min. It can be concluded that lemon, a natural product which is easily obtained, acts as a biocide against V. cholerae, and is, therefore, an efficient decontaminant, harmless to humans.

Serum interleukin-6 levels in murine models of Candida albicans infection.

PubMed

Kovács, Renátó; Czudar, Anita; Horváth, László; Szakács, Levente; Majoros, László; Kónya, József

2014-03-01

Two Balb/C mouse models of Candida infection were used to detect serum interleukin-6 (IL-6) responses. The first model used systemic infection by Candida albicans ATCC 10231 strain infected through the lateral tail vein of mice without any specific pretreatment. The median Candida burdens of the kidneys were 1.5 × 106 CFU/ml 24 h postinoculation (p.i.) and 1.2 × 107 CFU/ml 72 h p.i., while median serum IL-6 levels were 479.3 pg/ml and 934.5 pg/ml, respectively. The Candida burden showed significant correlation with serum IL-6 24 h p.i. (R2 = 0.6358; P = 0.0082) but not 72 h p.i.The second model was a mouse vaginitis model applying intravaginal inoculation of mice pretreated with subcutaneous estradiol-valerate (10 mg/ml) 3 days before infection. Candida cell count in vaginal lavage fluid was 2.8 × 106 CFU/ml 24 h p.i. and 1.4 × 108 CFU/ml 72 h p.i. Serum IL-6 response was detected in 4 of 15 mice 24 h p.i. and 9 of 15 mice 72 h p.i. Even the responders had low IL-6 serum levels (mean values 29.9 pg/ml and 60.1 pg/ml, respectively) not correlating with Candida cell count in vaginal lavage fluid.In conclusion, serum IL-6 had strong relationship with systemic C. albicans infection while the local C. albicans infection of the vagina led to partial, prolonged and limited serum IL-6 response.

[Detection of Brucella with an automatic hemoculture system: Bact/Alert].

PubMed

Casas, J; Partal, Y; Llosá, J; Leiva, J; Navarro, J M; de la Rosa, M

1994-12-01

The ability of in vitro and in vivo detection of Brucella spp. with the Bact/Alert system was studied. Three strains of Brucella melitensis and two of Brucella abortus were used. Different dilutions of the five strains were performed in trypticase soy broth (TSB), achieving concentrations of 1 cfu/ml, 5 cfu/ml, 10 cfu/ml and 100 cfu/ml. Ten ml of each dilution and strain were inoculated into 5 aerobic bottles Bact/Alert and 5 biphasic Hemóline bottles. Furthermore, over a 9 month period, 8,216 bottles of Bact/Alert bottles from hospitalized patients and from the emergency department were processed in the authors' laboratory. The mean detection time for Brucella growth was from 2 to 3 days with the Bact/Alert system, and 14 days in the biphasic bottles. Former bottles processed in the authors' laboratory, 11 aerobic bottles belonged to 5 patients in whom brucelosis was confirmed by bloodculture. The Bact/Alert system detected Brucella melitensis in only on bottle at 2.9 days of incubation. In 7 bottles Bact/Alert detected B. melitensis by a blind pass of these bottles at 10 to 20 days of incubation. These results suggest that the Bact/Alert system does not totally solve the diagnosis of brucellosis. Blind passes of the bloodcultures are required.

In vitro and in vivo activity of a killer peptide against Malassezia pachydermatis causing otitis in dogs.

PubMed

Cafarchia, Claudia; Immediato, Davide; Paola, Giancarlo Di; Magliani, Walter; Ciociola, Tecla; Conti, Stefania; Otranto, Domenico; Polonelli, Luciano

2014-05-01

In order to overcome the limitations inherent in current pharmacological treatments for Malassezia pachydermatis, the cause of otitis externa in dogs, the efficacy of a killer decapeptide (KP) was evaluated in vitro and in vivo. Sixteen dogs with naturally occurring M. pachydermatis otitis externa were enrolled, and the in vitro fungicidal activity of KP was evaluated using yeasts recovered from these animals. The therapeutic activity was evaluated in four groups of four animals each. The dogs were topically treated with KP (150 μl, 2 mg/ml) three times per week (group A) or every day (group B), treated with a scramble peptide every day (group C), or left untreated (group D). Assessment of clinical signs (pruritus, erythema, and lichenification and/or hyperpigmentation), expressed as mean of the total clinical index score (mTCIS), the population size of M. pachydermatis at the cytological examination (mean number of yeast cells at 40× magnification [mYC]), and culture testing (mean number of log10 CFU/swab [mCFU]), were conducted daily from the first day of treatment (T0) until two consecutive negative cultures (mCFU ≤ 2). KP showed an in vitro fungicidal effect against M. pachydermatis isolates, with an MFC90 value of 1 μg/ml. The mTCIS, mYC and mCFU were negative only in animals in group B after T8. Daily administration of KP for 8 days was safe and effective in controlling both clinical signs and the population size of M. pachydermatis causing otitis externa, thus offering an alternative to the currently available therapeutic or prophylactic protocols for recurrent cases of Malassezia otitis in dogs.

In vitro pharmacodynamics of human simulated exposures of ceftaroline and daptomycin against MRSA, hVISA, and VISA with and without prior vancomycin exposure.

PubMed

Bhalodi, Amira A; Hagihara, Mao; Nicolau, David P; Kuti, Joseph L

2014-01-01

The effects of prior vancomycin exposure on ceftaroline and daptomycin therapy against methicillin-resistant Staphylococcus aureus (MRSA) have not been widely studied. Humanized free-drug exposures of vancomycin at 1 g every 12 h (q12h), ceftaroline at 600 mg q12h, and daptomycin at 10 mg/kg of body weight q24h were simulated in a 96-h in vitro pharmacodynamic model against three MRSA isolates, including one heteroresistant vancomycin-intermediate S. aureus (hVISA) isolate and one VISA isolate. A total of five regimens were tested: vancomycin, ceftaroline, and daptomycin alone for the entire 96 h, and then sequential therapy with vancomycin for 48 h followed by ceftaroline or daptomycin for 48 h. Microbiological responses were measured by the changes in log10 CFU during 96 h from baseline. Control isolates grew to 9.16 ± 0.32, 9.13 ± 0.14, and 8.69 ± 0.28 log10 CFU for MRSA, hVISA, and VISA, respectively. Vancomycin initially achieved ≥3 log10 CFU reductions against the MRSA and hVISA isolates, followed by regrowth beginning at 48 h; minimal activity was observed against VISA. The change in 96-h log10 CFU was largest for sequential therapy with vancomycin followed by ceftaroline (-5.22 ± 1.2, P = 0.010 versus ceftaroline) and for sequential therapy with vancomycin followed by ceftaroline (-3.60 ± 0.6, P = 0.037 versus daptomycin), compared with daptomycin (-2.24 ± 1.0), vancomycin (-1.40 ± 1.8), and sequential therapy with vancomycin followed by daptomycin (-1.32 ± 1.0, P > 0.5 for the last three regimens). Prior exposure of vancomycin at 1 g q12h reduced the initial microbiological response of daptomycin, particularly for hVISA and VISA isolates, but did not affect the response of ceftaroline. In the scenario of poor vancomycin response for high-inoculum MRSA infection, a ceftaroline-containing regimen may be preferred.

Effects of Anticoagulant, Processing Delay, and Assay Method (Branched DNA versus Reverse Transcriptase PCR) on Measurement of Human Immunodeficiency Virus Type 1 RNA Levels in Plasma

PubMed Central

Kirstein, Lynn M.; Mellors, John W.; Rinaldo, Charles R.; Margolick, Joseph B.; Giorgi, Janis V.; Phair, John P.; Dietz, Edith; Gupta, Phalguni; Sherlock, Christopher H.; Hogg, Robert; Montaner, J. S. G.; Muñoz, Alvaro

1999-01-01

We conducted two studies to determine the potential influence of delays in blood processing, type of anticoagulant, and assay method on human immunodeficiency virus type 1 (HIV-1) RNA levels in plasma. The first was an experimental study in which heparin- and EDTA-anticoagulated blood samples were collected from 101 HIV-positive individuals and processed to plasma after delays of 2, 6, and 18 h. HIV-1 RNA levels in each sample were then measured by both branched-DNA (bDNA) and reverse transcriptase PCR (RT-PCR) assays. Compared to samples processed within 2 h, the loss (decay) of HIV-1 RNA in heparinized blood was significant (P < 0.05) but small after 6 h (bDNA assay, −0.12 log10 copies/ml; RT-PCR, −0.05 log10 copies/ml) and after 18 h (bDNA assay, −0.27 log10 copies/ml; RT-PCR, −0.15 log10 copies/ml). Decay in EDTA-anticoagulated blood was not significant after 6 h (bDNA assay, −0.002 log10 copies/ml; RT-PCR, −0.02 log10 copies/ml), but it was after 18 h (bDNA assay, −0.09 log10 copies/ml; RT-PCR, −0.09 log10 copies/ml). Only 4% of samples processed after 6 h lost more than 50% (≥0.3 log10 copies/ml) of the HIV-1 RNA, regardless of the anticoagulant or the assay that was used. The second study compared HIV-1 RNA levels in samples from the Multicenter AIDS Cohort Study (MACS; samples were collected in heparin-containing tubes in 1985, had a 6-h average processing delay, and were assayed by bDNA assay) and the British Columbia Drug Treatment Program (BCDTP) (collected in EDTA- or acid citrate dextrose-containing tubes in 1996 and 1997, had a 2-h maximum processing delay, and were assayed by RT-PCR). HIV-1 RNA levels in samples from the two cohorts were not significantly different after adjusting for CD4+-cell count and converting bDNA assay values to those corresponding to the RT-PCR results. In summary, the decay of HIV-1 RNA measured in heparinized blood after 6 h was small (−0.05 to −0.12 log10 copies/ml), and the minor impact of this decay on HIV-1 RNA concentrations in archived plasma samples of the MACS was confirmed by the similarity of CD4+-cell counts and assay-adjusted HIV-1 RNA concentrations in the MACS and BCDTP. PMID:10405379

Behavior of Escherichia coli O157:H7 and Listeria monocytogenes during fermentation and storage of camel yogurt.

PubMed

Al-Nabulsi, Anas A; Olaimat, Amin N; Osaili, Tareq M; Ayyash, Mutamed M; Abushelaibi, Aisha; Jaradat, Ziad W; Shaker, Reyad; Al-Taani, Mahmoud; Holley, Richard A

2016-03-01

In addition to its nutritional and therapeutic properties, camel milk has the ability to suppress the growth of a wide range of foodborne pathogens, but there is a lack of information regarding the behavior of these pathogens in products such as yogurt produced from camel milk. The objective of the current study was to investigate the behavior of Listeria monocytogenes and Escherichia coli O157:H7 during manufacture and storage of camel yogurt. Camel milk inoculated with L. monocytogenes and E. coli O157:H7 was fermented at 43° C for 5h using freeze-dried lactic acid bacteria (LAB) starter cultures (Streptococcus thermophilus and Lactobacillus bulgaricus) and stored at 4 or 10 °C for 14 d. Camel milk inoculated with L. monocytogenes and E. coli O157:H7 without starter culture was also prepared. During fermentation, the numbers of L. monocytogenes and E. coli O157:H7 increased 0.3 and 1.6 log cfu/mL, respectively, in the presence of LAB, and by 0.3 and 2.7 log cfu/mL in the absence of LAB. During storage at 4 or 10 °C, L. monocytogenes increased 0.8 to 1.2 log cfu/mL by 14 d in camel milk without LAB, but in the presence of LAB, the numbers of L. monocytogenes were reduced by 1.2 to 1.7 log cfu/mL by 14 d. Further, E. coli O157:H7 numbers in camel milk were reduced by 3.4 to 3.5 log cfu/mL in the absence of LAB, but E. coli O157:H7 was not detected (6.3 log cfu/mL reduction) by 7d in camel yogurt made with LAB and stored at either temperature. Although camel milk contains high concentrations of natural antimicrobials, L. monocytogenes was able to tolerate these compounds in camel yogurt stored at refrigerator temperatures. Therefore, appropriate care should be taken during production of yogurt from camel milk to minimize the potential for postprocess contamination by this and other foodborne pathogens. Copyright © 2016 American Dairy Science Association. Published by Elsevier Inc. All rights reserved.

[MAXIMUM SINGLE DOSE OF COLLOIDAL SILVER NEGATIVELY AFFECTS ERYTHROPOIESIS IN VITRO].

PubMed

Tishevskayal, N V; Zakharovl, Y M; Bolotovl, A A; Arkhipenko, Yu V; Sazontova, T G

2015-01-01

Erythroblastic islets (EI) of rat bone marrow were cultured for 24 h in the presence of silver nanoparticles (1.07 · 10(-4) mg/ml; 1.07 · 10(-3) mg/ml; and 1.07 · 10(-2) mg/mL). The colloidal silver at 1.07 · 10(-3) mg/ml concentration inhibited the formation of new Elby disrupting contacts of bone marrow macrophages with CFU-E (erythropoiesis de novo) by 65.3% (p < 0.05). Colloidal silver nanoparticles suppressed the reconstruction of erythropoiesis and inhibited the formation of new EI by disrupting contacts of CFU-E and central macrophages with matured erythroidal "crown" (erythropoiesis de repeto). The colloidal silver concentration of 1.07 · 10(-3) mg/ml in the culture medium also reduced the number of self-reconstructing EI by 67.5% (p <0.05), whereas 1.07 · 10(-2) mg/ml colloidal silver reduced this value by 93.7% (p < 0.05). Silver nanoparticles retarded maturation of erythroid cells at the stage of oxiphylic normoblast denucleation: 1.07 · 10(-3) mg/ml colloidal silver increased the number of mature El by 53% (p < 0.05). The retardation of erythropoiesis by colloidal silver in concentration equivalent to the maximum single dose is related to the effect of silver nanoparticles rather than glycerol present in the colloidal suspension.

Differential Expression of Virulence Genes and Motility in Ralstonia (Pseudomonas) solanacearum during Exponential Growth.

PubMed

Clough, S J; Flavier, A B; Schell, M A; Denny, T P

1997-03-01

A complex network regulates virulence in Ralstonia solanacearum (formerly Pseudomonas solanacearum); central to this system is PhcA, a LysR-type transcriptional regulator. We report here that two PhcA-regulated virulence factors, endoglucanase (Egl) and acidic exopolysaccharide I (EPS I), and motility are expressed differentially during exponential growth in batch cultures. Tests with strains carrying lacZ fusions in a wild-type genetic background revealed that expression (on a per-cell basis) of phcA was constant but expression of egl and epsB increased 20- to 50-fold during multiplication from 1 x 10(sup7) to 5 x 10(sup8) CFU/ml. Expression of xpsR, an intermediate regulator downstream of PhcA in the regulatory cascade for eps expression, was similar to that of epsB and egl. Motility track photography revealed that all strains were essentially nonmotile at 10(sup6) CFU/ml. As cell density increased, 30 to 50% of wild-type cells were motile between 10(sup7) and 10(sup8) CFU/ml, but this population was again nonmotile at 10(sup9) CFU/ml. In contrast, about 60% of the cells of phcB and phcA mutants remained motile at 10(sup9) CFU/ml. Expression of phcB, which is not positively regulated by PhcA, was the inverse of epsB, egl, and xpsR (i.e., it decreased 20-fold at high cell density). PhcB is essential for production of an extracellular factor, tentatively identified as 3-hydroxypalmitic acid methyl ester (3-OH PAME), that might act as an exponential-phase signal to activate motility or expression of virulence genes. However, growth of the lacZ fusion strains in medium containing excess 3-OH PAME did not result in motility or expression of virulence genes at dramatically lower cell densities, suggesting that 3-OH PAME is not the only factor controlling these traits.

Actual distribution of Cronobacter spp. in industrial batches of powdered infant formula and consequences for performance of sampling strategies.

PubMed

Jongenburger, I; Reij, M W; Boer, E P J; Gorris, L G M; Zwietering, M H

2011-11-15

The actual spatial distribution of microorganisms within a batch of food influences the results of sampling for microbiological testing when this distribution is non-homogeneous. In the case of pathogens being non-homogeneously distributed, it markedly influences public health risk. This study investigated the spatial distribution of Cronobacter spp. in powdered infant formula (PIF) on industrial batch-scale for both a recalled batch as well a reference batch. Additionally, local spatial occurrence of clusters of Cronobacter cells was assessed, as well as the performance of typical sampling strategies to determine the presence of the microorganisms. The concentration of Cronobacter spp. was assessed in the course of the filling time of each batch, by taking samples of 333 g using the most probable number (MPN) enrichment technique. The occurrence of clusters of Cronobacter spp. cells was investigated by plate counting. From the recalled batch, 415 MPN samples were drawn. The expected heterogeneous distribution of Cronobacter spp. could be quantified from these samples, which showed no detectable level (detection limit of -2.52 log CFU/g) in 58% of samples, whilst in the remainder concentrations were found to be between -2.52 and 2.75 log CFU/g. The estimated average concentration in the recalled batch was -2.78 log CFU/g and a standard deviation of 1.10 log CFU/g. The estimated average concentration in the reference batch was -4.41 log CFU/g, with 99% of the 93 samples being below the detection limit. In the recalled batch, clusters of cells occurred sporadically in 8 out of 2290 samples of 1g taken. The two largest clusters contained 123 (2.09 log CFU/g) and 560 (2.75 log CFU/g) cells. Various sampling strategies were evaluated for the recalled batch. Taking more and smaller samples and keeping the total sampling weight constant, considerably improved the performance of the sampling plans to detect such a type of contaminated batch. Compared to random sampling, stratified random sampling improved the probability to detect the heterogeneous contamination. Copyright © 2011 Elsevier B.V. All rights reserved.

Inactivation of Heat Adapted and Chlorine Adapted Listeria Monocytogenes ATCC 7644 on Tomatoes Using Sodium Dodecyl Sulphate, Levulinic Acid and Sodium Hypochlorite Solution.

PubMed

Ijabadeniyi, Oluwatosin Ademola; Mnyandu, Elizabeth

2017-04-13

The effectiveness of sodium dodecyl sulphate (SDS), sodium hypochlorite solution and levulinic acid in reducing the survival of heat adapted and chlorine adapted Listeria monocytogenes ATCC 7644 was evaluated. The results against heat adapted L. monocytognes revealed that sodium hypochlorite solution was the least effective, achieving log reduction of 2.75, 2.94 and 3.97 log colony forming unit (CFU)/mL for 1, 3 and 5 minutes, respectively. SDS was able to achieve 8 log reduction for both heat adapted and chlorine adapted bacteria. When used against chlorine adapted L. monocytogenes sodium hypochlorite solution achieved log reduction of 2.76, 2.93 and 3.65 log CFU/mL for 1, 3 and 5 minutes, respectively. Using levulinic acid on heat adapted bacteria achieved log reduction of 3.07, 2.78 and 4.97 log CFU/mL for 1, 3, 5 minutes, respectively. On chlorine adapted bacteria levulinic acid achieved log reduction of 2.77, 3.07 and 5.21 log CFU/mL for 1, 3 and 5 minutes, respectively. Using a mixture of 0.05% SDS and 0.5% levulinic acid on heat adapted bacteria achieved log reduction of 3.13, 3.32 and 4.79 log CFU/mL for 1, 3 and 5 minutes while on chlorine adapted bacteria it achieved 3.20, 3.33 and 5.66 log CFU/mL, respectively. Increasing contact time also increased log reduction for both test pathogens. A storage period of up to 72 hours resulted in progressive log reduction for both test pathogens. Results also revealed that there was a significant difference (P≤0.05) among contact times, storage times and sanitizers. Findings from this study can be used to select suitable sanitizers and contact times for heat and chlorine adapted L. monocytogenes in the fresh produce industry.

Identification of lactic acid bacteria isolated from wine using real-time PCR.

PubMed

Kántor, Attila; Kluz, Maciej; Puchalski, Czeslaw; Terentjeva, Margarita; Kačániová, Miroslava

2016-01-01

Different lactic acid bacteria strains have been shown to cause wine spoilage, including the generation of substances undesirable for the health of wine consumers. The aim of this study was to investigate the occurrence of selected species of heterofermentative lactobacilli, specifically Lactobacillus brevis, Lactobacillus hilgardii, and Lactobacillus plantarum in six different Slovak red wines following the fermentation process. In order to identify the dominant Lactobacillus strain using quantitative (real time) polymerized chain reaction (qPCR) method, pure lyophilized bacterial cultures from the Czech Collection of Microorganisms were used. Six different red wine samples following malolactic fermentation were obtained from selected wineries. After collection, the samples were subjected to a classic plate dilution method for enumeration of lactobacilli cells. Real-time PCR was performed after DNA extraction from pure bacterial strains and wine samples. We used SYBR® Green master mix reagents for measuring the fluorescence in qPCR. The number of lactobacilli ranged from 3.60 to 5.02 log CFU mL(-1). Specific lactobacilli strains were confirmed by qPCR in all wine samples. The number of lactobacilli ranged from 10(3) to 10(6) CFU mL(-1). A melting curve with different melting temperatures (T(m)) of DNA amplicons was obtained after PCR for the comparison of T(m) of control and experimental portions, revealing that the most common species in wine samples was Lactobacillus plantarum with a T(m) of 84.64°C.

Escherichia coli and Cronobacter sakazakii in 'Tommy Atkins' minimally processed mangos: Survival, growth and effect of UV-C and electrolyzed water.

PubMed

Santo, David; Graça, Ana; Nunes, Carla; Quintas, Célia

2018-04-01

These studies were aimed at assessing the growing capacity of Escherichia coli and Cronobacter sakazakii and the effectiveness of Ultraviolet-C (UV-C) radiation, acidic electrolyzed (AEW) and neutral electrolyzed (NEW) waters in the inhibition of these bacteria on minimally processed 'Tommy Atkins' mangoes (MPM). The fruits were contaminated by dip inoculation and kept 10 days at 4, 8, 12 and 20 °C while enumerating bacteria. Contaminated mangoes were disinfected using UV-C (2.5, 5, 7.5 and 10 kJ/m 2 ), AEW, NEW and sodium hypochlorite (SH) and the microorganisms were monitored. None of the enterobacteria grew at 4, 8 and 12 °C regardless of having persisted during the 10-day period. At 20 °C, E. coli and C. sakazakii grew, after adaption phases of 48 h and 24 h, to values of 8.7 and 8.5 log cfu/g at day eight, respectively. E. coli showed the highest reduction counts on the MPM washed with NEW and SH (2.2 log cfu/g). UV-C was more effective in reducing C. sakazakii (2.4-2.6 log cfu/g), when compared to AEW, NEW and SH (1.2-1.8 log cfu/g). The efficacy of decontamination technologies depends on microorganisms, highlighting the importance of preventing contamination at the primary production and of combining different methods to increase the safety of fresh-cut fruits. Copyright © 2017 Elsevier Ltd. All rights reserved.

Refrigerated Shelf Life of a Coconut Water-Oatmeal Mix and the Viability of Lactobacillus Plantarum Lp 115-400B

PubMed Central

Dharmasena, Muthu; Barron, Felix; Fraser, Angela; Jiang, Xiuping

2015-01-01

Non-dairy probiotic products have the advantage of being lactose-free and can be manufactured to sustain the growth of probiotics. In this study, coconut water and oatmeal were used with the probiotic, Lactobacillus plantarum Lp 115-400B (L. plantarum) as a starter culture. Two separate treatments were carried out probiotic (P) and probiotic and prebiotic (PP) added. In both treatments, oatmeal-coconut water matrix was inoculated with 7 log CFU/g of L. plantarum and fermented at 27 °C for 10 h. For the PP treatment, 1 g of inulin/100 mL of the product was added additionally. The fermented products were then refrigerated (4 °C) and the viability of L. plantarum, pH, total acidity, and apparent viscosity of the matrix were monitored at selected time intervals. The shelf life to reach was defined by maintenance of L. plantarum count of 7 log CFU/g product. Refrigerated shelf life was determined to be seven-weeks for the P treatment and five-weeks for PP treatment. A significant reduction of pH was observed at the end of the considered shelf life; conversely, the apparent viscosity of the product did not change significantly. PMID:28231208

Refrigerated Shelf Life of a Coconut Water-Oatmeal Mix and the Viability of Lactobacillus Plantarum Lp 115-400B.

PubMed

Dharmasena, Muthu; Barron, Felix; Fraser, Angela; Jiang, Xiuping

2015-08-10

Non-dairy probiotic products have the advantage of being lactose-free and can be manufactured to sustain the growth of probiotics. In this study, coconut water and oatmeal were used with the probiotic, Lactobacillus plantarum Lp 115-400B ( L. plantarum ) as a starter culture. Two separate treatments were carried out probiotic (P) and probiotic and prebiotic (PP) added. In both treatments, oatmeal-coconut water matrix was inoculated with 7 log CFU/g of L. plantarum and fermented at 27 °C for 10 h. For the PP treatment, 1 g of inulin/100 mL of the product was added additionally. The fermented products were then refrigerated (4 °C) and the viability of L. plantarum , pH, total acidity, and apparent viscosity of the matrix were monitored at selected time intervals. The shelf life to reach was defined by maintenance of L. plantarum count of 7 log CFU/g product. Refrigerated shelf life was determined to be seven-weeks for the P treatment and five-weeks for PP treatment. A significant reduction of pH was observed at the end of the considered shelf life; conversely, the apparent viscosity of the product did not change significantly.

Photodynamic therapy with high-power LED mediated by erythrosine eliminates Enterococcus faecalis in planktonic forms.

PubMed

Borba, Alberto Sabin Moura; da Silva Pereira, Sângela Maria; Borba, Mellyna Cavalcante Mendes; Paschoal, Marco Aurélio Benini; de Jesus Tavarez, Rudys Rodolfo; de Castro Rizzi, Claudia; Ferreira, Meire Coelho; Maia Filho, Etevaldo Matos

2017-09-01

The failure of endodontic treatment is linked to the presence of microorganisms, particularly Enterococcus faecalis, in the root canals. This study evaluated the effectiveness of photodynamic therapy (PDT) using erythrosine irradiated by a high-power curing light on a planktonic suspension culture of E. faecalis. Bacterial suspensions of E. faecalis were adjusted and then mixed in a 1:1 proportion, in triplicate, in treatment groups by varying the length of irradiation time (120 and 240s) and the molarity of the erythrosine (5 and 10μM). In order to verify the post-treatment bactericidal effect, a count of the viable bacteria was performed (CFUmL -1 ) and transformed into Log10 CFU. The one-way ANOVA with Tukey post-hoc test was applied to check for differences between the groups. The bacteria were completely eradicated in the groups that used PDT with 5μM 240s, 10μM 120s and 10μM 240s (p≪0.001). The effect of the PDT 5μM 120s group was significant (p≪0.05) in comparison with the groups using only light or only erythrosine. Positive control (exposure to 2.5% NaClO for 120 and 240s) completely eradicated E. faecalis. The negative control (PBS) did not alter the quantities of E. faecalis CFU with 9.605 Log10 CFU at 120s and 9.621 Log10 CFU at 240s. PDT with erythrosine in a concentration of 10μM and high-power LED is capable of totally eliminating E. faecalis in planktonic suspension. Copyright © 2017 Elsevier B.V. All rights reserved.

Survival and Heat Resistance of Listeria monocytogenes after Exposure to Alkali and Chlorine

PubMed Central

Taormina, P. J.; Beuchat, L. R.

2001-01-01

A strain of Listeria monocytogenes isolated from a drain in a food-processing plant was demonstrated, by determination of D values, to be more resistant to the lethal effect of heat at 56 or 59°C following incubation for 45 min in tryptose phosphate broth (TPB) at pH 12.0 than to that of incubation for the same time in TPB at pH 7.3. Cells survived for at least 6 days when they were suspended in TPB at pHs 9.0, 10.0, and 11.0 and stored at 4 or 21°C. Cells of L. monocytogenes incubated at 37°C for 45 min and then stored for 48 or 144 h in TPB at pH 10.0 were more resistant to heat treatment at 56°C than were cells stored in TPB at pH 7.3. The alkaline-stress response in L. monocytogenes may induce resistance to otherwise lethal thermal-processing conditions. Treatment of cells in 0.05 M potassium phosphate buffer (pH 7.00 ± 0.05) containing 2.0 or 2.4 mg of free chlorine per liter reduced populations by as much as 1.3 log10 CFU/ml, while treatment with 6.0 mg of free chlorine per liter reduced populations by as much as 4.02 log10 CFU/ml. Remaining subpopulations of chlorine-treated cells exhibited some injury, and cells treated with chlorine for 10 min were more sensitive to heating at 56°C than cells treated for 5 min. Contamination of foods by L. monocytogenes cells that have survived exposure to processing environments ineffectively cleaned or sanitized with alkaline detergents or disinfectants may have more severe implications than previously recognized. Alkaline-pH-induced cross-protection of L. monocytogenes against heat has the potential to enhance survival in minimally processed as well as in heat-and-serve foods and in foods on holding tables, in food service facilities, and in the home. Cells surviving exposure to chlorine, in contrast, are more sensitive to heat; thus, the effectiveness of thermal processing in achieving desired log10-unit reductions is not compromised in these cells. PMID:11375163

Antimicrobial effects of hypochlorite on Escherichia coli in water and selected vegetables.

PubMed

Erkmen, Osman

2010-08-01

In this study, the antimicrobial effects of hypochlorite (HOCl) on Escherichia coli in tap water were investigated. The effects of 0.1% thyme oil and 100 mg/L HOCl on E. coli on vegetables (lettuce, parsley leafs, and red pepper) were also studied. E. coli was reduced by 2.54, 3.33, 3.93, 4.87, and 5.57 log colony forming units (cfu)/mL with 0.25, 0.5, 1.0, 10, and 50 mg/mL HOCl, respectively. There was an increase of more than 30% in the inactivation of E. coli with 10 degrees C rise in temperature, a remarkable increase in antimicrobial activity at pH 5.0 was also observed with 5.62 log cfu/mL reductions in 30 sec, as well as marked neutralization of the effect in the presence of 0.1% peptone in water was noted. Biphasic kinetics in the inactivation curves of E. coli was observed. HOCl, thyme oil, and their mixture reduced the number of E. coli between 1.23 and 3.75 log cfu/mL after 5-min exposure on vegetables. The degree of E. coli inactivation depends on concentration of residual chlorine, suspending medium, type of vegetables, and the use of thyme essential oil.

Slurry-phase biodegradation of weathered oily sludge waste.

PubMed

Machín-Ramírez, C; Okoh, A I; Morales, D; Mayolo-Deloisa, K; Quintero, R; Trejo-Hernández, M R

2008-01-01

We assessed the biodegradation of a typical oily sludge waste (PB401) in Mexico using several regimes of indigenous microbial consortium and relevant bioremediation strategies in slurry-phase system. Abiotic loss of total petroleum hydrocarbons (TPH) in the PB401 was insignificant, and degradation rates under the various treatment conditions ranged between 666.9 and 2168.7 mg kg(-1) day(-1) over a 15 days reaction period, while viable cell count peaked at between log(10)5.7 and log(10)7.4 cfu g(-1). Biostimulation with a commercial fertilizer resulted in 24% biodegradation of the TPH in the oily waste and a corresponding peak cell density of log(10)7.4 cfu g(-1). Addition of non-indigenous adapted consortium did not appear to enhance the removal of TPH from the oily waste. It would appear that the complexities of the components of the alkylaromatic fraction of the waste limited biodegradation rate even in a slurry system.

Cell division inhibitors with efficacy equivalent to isoniazid in the acute murine Mycobacterium tuberculosis infection model

PubMed Central

Knudson, Susan E.; Awasthi, Divya; Kumar, Kunal; Carreau, Alexandra; Goullieux, Laurent; Lagrange, Sophie; Vermet, Hélène; Ojima, Iwao; Slayden, Richard A.

2015-01-01

Objectives The increasing number of clinical strains resistant to one or more of the front-line TB drugs complicates the management of this disease. To develop next-generation benzimidazole-based FtsZ inhibitors with improved efficacy, we employed iterative optimization strategies based on whole bacteria potency, bactericidal activity, plasma and metabolic stability and in vivo efficacy studies. Methods Candidate benzimidazoles were evaluated for potency against Mycobacterium tuberculosis H37Rv and select clinical strains, toxicity against Vero cells and compound stability in plasma and liver microsomes. The efficacy of lead compounds was assessed in the acute murine M. tuberculosis infection model via intraperitoneal and oral routes. Results MICs of SB-P17G-A33, SB-P17G-A38 and SB-P17G-A42 for M. tuberculosis H37Rv and select clinical strains were 0.18–0.39 mg/L. SB-P17G-A38 and SB-P17G-A42 delivered at 50 mg/kg twice daily intraperitoneally or orally demonstrated efficacy in reducing the bacterial load by 5.7–6.3 log10 cfu in the lungs and 3.9–5.0 log10 cfu in the spleen. SB-P17G-A33 delivered at 50 mg/kg twice daily intraperitoneally or orally also reduced the bacterial load by 1.7–2.1 log10 cfu in the lungs and 2.5–3.4 log10 cfu in the spleen. Conclusions Next-generation benzimidazoles with excellent potency and efficacy against M. tuberculosis have been developed. This is the first report on benzimidazole-based FtsZ inhibitors showing an equivalent level of efficacy to isoniazid in an acute murine M. tuberculosis infection model. PMID:26245639

Effect of chlorine treatment on inhibition of E. coli serogroup O2 incorporation into 7-day-old biofilm on polyvinylchloride surface.

PubMed

Maharjan, P; Dey, S; Huff, G; Zhang, W; Phillips, G K; Watkins, S

2017-08-01

Poultry waterlines are constructed using polyvinylchloride (PVC) material on which bacterial biofilm can easily form. Biofilm can harbor pathogens including avian pathogenic E. coli (APEC) strains. An in vitro evaluation was performed to determine if E. coli sero group O2 (avian pathogenic) could attach on a PVC surface that had pre-formed biofilm and if this phenomenon could be affected when water was treated with chlorine. Initially, biofilm growth was induced in PVC test coupons (15.16 cm2) for a 7-day period mimicking the waterline scenario in the first wk of poultry brooding; and then this biofilm was challenged with E. coli O2 seeded water in presence/absence of chlorine treatment. After rinsing, test coupons were sampled for bacterial (APC) and E. coli O2 enumeration at various occasions post seeding the pathogen and chlorine treatment. Day 7 APC recovered from coupons was 4.35 log10 cfu/cm2 in trial 1 and 3.66 log10 cfu/cm2 in trial 2. E. coli O2 was not recovered from chlorine treated test coupons (P < 0.05), whereas it was retrieved from untreated coupons (untreated contained > 3 log10 cfu/cm2 in trial 1 and > 2 log10 cfu/cm2 in trial 2). This study suggests that E. coli O2 can incorporate into pre-formed biofilm on a PVC surface within 24 h if water sanitation is not present, and the attachment time of the pathogen can prolong in the absence of already formed biofilm. © 2017 Poultry Science Association Inc.

Sensitizing Clostridium difficile Spores with Germinants on Skin and Environmental Surfaces Represents a New Strategy for Reducing Spores via Ambient Mechanisms

PubMed Central

Nerandzic, Michelle M.; Donskey, Curtis J.

2017-01-01

Background Clostridium difficile is a leading cause of healthcare-associated infections worldwide. Prevention of C. difficile transmission is challenging because spores are not killed by alcohol-based hand sanitizers or many commonly used disinfectants. One strategy to control spores is to induce germination, thereby rendering the spores more susceptible to benign disinfection measures and ambient stressors. Methods/Results C. difficile spores germinated on skin after a single application of cholic acid-class bile salts and co-germinants; for 4 C. difficile strains, recovery of viable spores from skin was reduced by ~0.3 log10CFU to 2 log10CFU after 2 hours and ~1 log10CFU to > 2.5 log10CFU after 24 hours. The addition of taurocholic acid to 70% and 30% ethanol significantly enhanced reduction of viable spores on skin and on surfaces. Desiccation, and to a lesser extent the presence of oxygen, were identified as the stressors responsible for reductions of germinated spores on skin and surfaces. Additionally, germinated spores became susceptible to killing by pH 1.5 hydrochloric acid, suggesting that germinated spores that remain viable on skin and surfaces might be killed by gastric acid after ingestion. Antibiotic-treated mice did not become colonized after exposure to germinated spores, whereas 100% of mice became colonized after exposure to the same quantity of dormant spores. Conclusions Germination could provide a new approach to reduce C. difficile spores on skin and in the environment and to render surviving spores less capable of causing infection. Our findings suggest that it may be feasible to develop alcohol-based hand sanitizers containing germinants that reduce spores on hands. PMID:29167835

Microbiological Evaluation of the Efficacy of Soapy Water to Clean Hands: A Randomized, Non-Inferiority Field Trial

PubMed Central

Amin, Nuhu; Pickering, Amy J.; Ram, Pavani K.; Unicomb, Leanne; Najnin, Nusrat; Homaira, Nusrat; Ashraf, Sania; Abedin, Jaynal; Islam, M. Sirajul; Luby, Stephen P.

2014-01-01

We conducted a randomized, non-inferiority field trial in urban Dhaka, Bangladesh among mothers to compare microbial efficacy of soapy water (30 g powdered detergent in 1.5 L water) with bar soap and water alone. Fieldworkers collected hand rinse samples before and after the following washing regimens: scrubbing with soapy water for 15 and 30 seconds; scrubbing with bar soap for 15 and 30 seconds; and scrubbing with water alone for 15 seconds. Soapy water and bar soap removed thermotolerant coliforms similarly after washing for 15 seconds (mean log10 reduction = 0.7 colony-forming units [CFU], P < 0.001 for soapy water; mean log10 reduction = 0.6 CFU, P = 0.001 for bar soap). Increasing scrubbing time to 30 seconds did not improve removal (P > 0.05). Scrubbing hands with water alone also reduced thermotolerant coliforms (mean log10 reduction = 0.3 CFU, P = 0.046) but was less efficacious than scrubbing hands with soapy water. Soapy water is an inexpensive and microbiologically effective cleansing agent to improve handwashing among households with vulnerable children. PMID:24914003

Procedural revision to the use-dilution methods: establishment of maximum log density value for test microbes on inoculated carriers.

PubMed

Tomasino, Stephen F; Pines, Rebecca M; Hamilton, Gordon C

2012-01-01

(Staphylococcus aureus) and 964.02 (Pseudomonas aeruginosa), were revised in 2009 to include a standardized procedure to measure the log density of the test microbe and to establish a minimum mean log density value of 6.0 (geometric mean of 1.0 x 10(6) CFU/carrier) to qualify the test results. This report proposes setting a maximum mean log density value of 7.0 (geometric mean of 1.0 x 10(7) CFU/carrier) to further standardize the procedure. The minimum value was based on carrier count data collected by four laboratories over an 8-year period (1999-2006). The data have been updated to include an additional 4 years' worth of data (2006-2010) collected by the same laboratories. A total of 512 tests were conducted on products bearing claims against P. aeruginosa and S. aureus with and without an organic soil load (OSL) added to the inoculum (as specified on the product label claim). Six carriers were assayed in each test, for a total of 3072 carriers. Mean log densities for each of the 512 tests were at least 6.0. With the exception of two tests, one for P. aeruginosa without OSL and one for S. aureus with OSL, the mean log densities did not exceed 7.5 (geometric mean of 3.2 x 10(7) CFU/carrier). Across microbes and OSL treatments, the mean log density (+/- SEM) was 6.80 (+/- 0.07) per carrier (a geometric mean of 6.32 x 10(6) CFUlcarrier) and acceptable repeatability (0.28) and reproducibility (0.31) SDs were exhibited. A maximum mean log density per carrier of 7.0 is being proposed here as a validity requirement for S. aureus and P. aeruginosa. A modification to the method to allow for dilution of the final test cultures to achieve carrier counts within 6.0-7.0 logs is also being proposed. Establishing a range of 6.0-7.0 logs will help improve the reliability of the method and should allow for more consistent results within and among laboratories.

Biological efficacy and toxic effect of emergency water disinfection process based on advanced oxidation technology.

PubMed

Tian, Yiping; Yuan, Xiaoli; Xu, Shujing; Li, Rihong; Zhou, Xinying; Zhang, Zhitao

2015-12-01

An innovative and removable water treatment system consisted of strong electric field discharge and hydrodynamic cavitation based on advanced oxidation technologies was developed for reactive free radicals producing and waterborne pathogens eliminating in the present study. The biological efficacy and toxic effects of this advanced oxidation system were evaluated during water disinfection treatments. Bench tests were carried out with synthetic microbial-contaminated water, as well as source water in rainy season from a reservoir of Dalian city (Liaoning Province, China). Results showed that high inactivation efficiency of Escherichia coli (>5 log) could be obtained for synthetic contaminated water at a low concentration (0.5-0.7 mg L(-1)) of total oxidants in 3-10 s. The numbers of wild total bacteria (108 × 10(3) CFU mL(-1)) and total coliforms (260 × 10(2) MPN 100 mL(-1)) in source water greatly reduced to 50 and 0 CFU mL(-1) respectively after treated by the advanced oxidation system, which meet the microbiological standards of drinking water, and especially that the inactivation efficiency of total coliforms could reach 100%. Meanwhile, source water qualities were greatly improved during the disinfection processes. The values of UV254 in particular were significantly reduced (60-80%) by reactive free radicals. Moreover, the concentrations of possible disinfection by-products (formaldehyde and bromide) in treated water were lower than detection limits, indicating that there was no harmful effect on water after the treatments. These investigations are helpful for the ecotoxicological studies of advanced oxidation system in the treatments of chemical polluted water or waste water. The findings of this work suggest that the developed water treatment system is ideal in the acute phases of emergencies, which also could offer additional advantages over a wide range of applications in water pollution control.

Microbiological characterization of vegetables and their rhizosphere soil in Eastern Poland.

PubMed

Kłapeć, Teresa; Wójcik-Fatla, Angelina; Cholewa, Alicja; Cholewa, Grażyna; Dutkiewicz, Jacek

2016-12-23

The aim of this study was to investigate the bacteriological quality of 5 kinds of vegetables (lettuce, dill, radish, beetroot, carrot) and their rhizosphere soil, originating from conventional farms located in the Lublin Province of Eastern Poland. A total number of 35 samples of fresh vegetables (FV) taken immediately from soil, 35 samples of soil from rhizosphere of these vegetables (SR) and 35 samples of vegetables sold at retail in the markets in Lublin (VR) were examined. The samples were analysed for the content of: aerobic mesophilic bacteria (AMB) grown at 30°C and 37°C, Gram-negative bacteria of Enterobacteriaceae family, faecal coliform (FC) bacteria, Salmonella spp., and Clostridium perfringens. Median AMB values determined at 30°C for FV, SR and VR were 5.27, 5.00, and 5.00 log 10 CFU g -1 , respectively, being significantly greater compared to those recorded at 37°C. The exceeding of the threshold value of 6.0 log 10 CFU g -1 proposed by Gelosa (1998) was noted only in 5 FV samples grown at 30°C (14.3%), and in 3 FV samples grown at 37°C (8.6%). The threshold value was never exceeded in SR and VR samples. Median concentrations of Enterobacteriaceae determined for FV, SR and VR were 4.03, 3.87, and 3.04 log 10 CFU g -1 , respectively. Eleven species of Enterobacteriaceae were identified in the FV, SR and VR samples. The percent of samples containing Escherichia coli was greatest for VR (22.9%), smaller for FV (17.1%) and smallest for SR (5.7%). The median concentrations of the faecal coliform bacteria (FC), determined by culture at 44°C, were low, amounting to 1.000 log 10 CFU g -1 for FV and SR and 0.00 for VR. All examined vegetable and soil samples tested negative for the presence of Salmonella. The median concentrations of Clostridium perfringens were low, amounting to 0.00 log 10 CFU g -1 for all categories of samples. This bacterium was relatively common in soil samples with the prevalence of 40.0%, but very rare in vegetable samples (occurring in 5.7% of FV and in none of VR samples). In conclusion, the results of the present study generally indicate that the microbiological quality of Polish vegetables grown on conventional farms is satisfactory and safe for consumers.

Modeling the survival of Escherichia coli O157:H7, Listeria monocytogenes, and Salmonella Typhimurium during fermentation, drying, and storage of soudjouk-style fermented sausage.

PubMed

Hwang, Cheng-An; Porto-Fett, Anna C S; Juneja, Vijay K; Ingham, Steven C; Ingham, Barbara H; Luchansky, John B

2009-02-28

This study quantified and modeled the survival of Escherichia coli O157:H7, Listeria monocytogenes and Salmonella Typhimurium in soudjouk-style fermented sausage during fermentation, drying, and storage. Batter prepared from ground beef (20% fat), seasonings, starter culture, and dextrose was separately inoculated with a multi-strain mixture of each pathogen to an initial inoculum of ca. 6.5 log(10) CFU/g in the batter. The sausages were subsequently fermented at 24 degrees C with a relative humidity (RH) of 90% to 95% for 3 to 5 days to ca. pH 5.2, pH 4.9 or pH 4.6, then dried at 22 degrees C to a(w) 0.92, a(w) 0.89, or a(w) 0.86, respectively, and then stored at 4, 21, or 30 degrees C for up to 60 days. Lethality of the three pathogens was modeled as a function of pH, a(w) and/or storage temperature. During fermentation to pH 5.2 to pH 4.6, cell reductions ranged from 0 to 0.9 log(10) CFU/g for E. coli O157:H7, 0.1 to 0.5 log(10) CFU/g for L. monocytogenes, and 0 to 2.2 log(10) CFU/g for S. Typhimurium. Subsequent drying of sausages of pH 5.2 to pH 4.6 at 22 degrees C with 80% to 85% RH for 3 to 7 days to a(w) of 0.92 to a(w) 0.86 resulted in additional reductions that ranged from 0 to 3.5 log(10) CFU/g for E. coli O157:H7, 0 to 0.4 log(10) CFU/g for L. monocytogenes, and 0.3 to 2.4 log(10) CFU/g for S. Typhimurium. During storage at 4, 21, or 30 degrees C the reduction rates of the three pathogens were generally higher (p<0.05) in sausages with lower pH and lower a(w) that were stored at higher temperatures. Polynomial equations were developed to describe the inactivation of the three pathogens during fermentation, drying, and storage. The applicability of the resulting models for fermented sausage was evaluated by comparing model predictions with published data. Pathogen reductions estimated by the models for E. coli O157:H7 and S. Typhimurium were comparable to 67% and 73% of published data, respectively. Due to limited published data for L. monocytogenes, the models for L. monocytogenes would need additional validations. Results of pathogen reductions from this study may be used as a reference to assist manufacturers of soudjouk-style sausages to adopt manufacturing processes that meet the regulatory requirements. The resulting models may also be used for estimating the survival of E. coli O157:H7 and S. Typhimurium in other similar fermented sausage during fermentation and storage.

Contamination of faecal coliforms in ice cubes sampled from food outlets in Kubang Kerian, Kelantan.

PubMed

Noor Izani, N J; Zulaikha, A R; Mohamad Noor, M R; Amri, M A; Mahat, N A

2012-03-01

The use of ice cubes in beverages is common among patrons of food outlets in Malaysia although its safety for human consumption remains unclear. Hence, this study was designed to determine the presence of faecal coliforms and several useful water physicochemical parameters viz. free residual chlorine concentration, turbidity and pH in ice cubes from 30 randomly selected food outlets in Kubang Kerian, Kelantan. Faecal coliforms were found in ice cubes in 16 (53%) food outlets ranging between 1 CFU/100mL to >50 CFU/ 100mL, while in the remaining 14 (47%) food outlets, in samples of tap water as well as in commercially bottled drinking water, faecal coliforms were not detected. The highest faecal coliform counts of >50 CFU/100mL were observed in 3 (10%) food outlets followed by 11-50 CFU/100mL and 1-10 CFU/100mL in 7 (23%) and 6 (20%) food outlets, respectively. All samples recorded low free residual chlorine concentration (<0.10mg/L) with the pH ranging between 5.5 and 7.3 and turbidity between 0.14-1.76 NTU. Since contamination by faecal coliforms was not detected in 47% of the samples, tap water and commercially bottled drinking water, it was concluded that (1) contamination by faecal coliforms may occur due to improper handling of ice cubes at the food outlets or (2) they may not be the water sources used for making ice cubes. Since low free residual chlorine concentrations were observed (<0.10mg/ L) in all samples as well as in both tap water and commercially bottled drinking water, with the pH ranged between 5.5-7.3, ineffective disinfection of water source as a contributing factor to such high counts of faecal coliforms in ice cubes also could not be ruled out. Therefore, a periodical, yet comprehensive check on the food outlets, including that of ice cube is crucial in ensuring better food and water for human consumption.

Efficacy of essential oils to reduce Salmonella in organic soil

USDA-ARS?s Scientific Manuscript database

Salmonella populations in soil were reduced by up to 5 log CFU/ml after 28 days of incubation using vinegar or eugenol. The bactericidal effect of Cinnamanaldehyde was not evident. S. negev was sensitive to oils resulting in significant reduction of this bacterium. Increase in oil concentration resu...

Bacteriophages reduce Yersinia enterocolitica contamination of food and kitchenware.

PubMed

Jun, Jin Woo; Park, Se Chang; Wicklund, Anu; Skurnik, Mikael

2018-04-20

Yersinia enterocolitica, the primary cause of yersiniosis, is one of the most important foodborne pathogens globally and is associated with the consumption of raw contaminated pork. In the current study, four virulent bacteriophages (phages), one of Podoviridae (fHe-Yen3-01) and three of Myoviridae (fHe-Yen9-01, fHe-Yen9-02, and fHe-Yen9-03), capable of infecting Y. enterocolitica were isolated and characterized. fHe-Yen9-01 had the broadest host range (61.3% of strains, 65/106). It demonstrated a latent period of 35 min and a burst size of 33 plaque-forming units/cell, and was found to have a genome of 167,773 bp with 34.79% GC content. To evaluate the effectiveness of phage fHe-Yen9-01 against Y. enterocolitica O:9 strain Ruokola/71, we designed an experimental model of the food market environment. Phage treatment after bacterial inoculation of food samples, including raw pork (4 °C, 72 h), ready-to-eat pork (26 °C, 12 h), and milk (4 °C, 72 h), prevented bacterial growth throughout the experiments, with counts decreasing by 1-3 logs from the original levels of 2-4 × 10 3  CFU/g or ml. Similarly, when artificially contaminated kitchen utensils, such as wooden and plastic cutting boards and knives, and artificial hands, were treated with phages for 2 h, bacterial growth was effectively inhibited, with counts decreasing by 1-2 logs from the original levels of ca 10 4  CFU/cm 2 or ml. To the best of our knowledge, this is the first report of the successful application of phages for the control of Y. enterocolitica growth in food and on kitchen utensils. Copyright © 2018 Elsevier B.V. All rights reserved.

Rationale behind high-dose amoxicillin therapy for acute otitis media due to penicillin-nonsusceptible pneumococci: support from in vitro pharmacodynamic studies.

PubMed Central

Lister, P D; Pong, A; Chartrand, S A; Sanders, C C

1997-01-01

To evaluate whether increased doses of amoxicillin should be used to treat acute pneumococcal otitis media, an in vitro pharmacokinetic model was used to evaluate the killing of pneumococci by amoxicillin when middle ear pharmacokinetics were simulated. Logarithmic-phase cultures were exposed to peak concentrations of 3, 6, and 9 microg of amoxicillin per ml every 12 h, and an elimination half-life of 1.6 h was simulated. Changes in viable bacterial counts were measured over 36 h. All three doses rapidly decreased the viable bacterial counts of penicillin-susceptible strains below the 10-CFU/ml limit of detection by 6 to 10 h and maintained counts below this limit through 36 h. The 3-microg/ml peak dose was much less effective against two of three strains with intermediate penicillin resistance and all three penicillin-resistant strains, with bacterial counts approaching those in drug-free control cultures by 12 h. The 6-microg/ml peak dose completely eliminated two of three strains with intermediate penicillin resistance and maintained viable counts of the other nonsusceptible strains at 1.5 to 2 logs below the initial inoculum through 36 h. The 9-microg/ml peak dose was most effective, completely eliminating all three strains with intermediate penicillin resistance and maintaining the viable counts of the resistant strains at 3 to 4 logs below the original inoculum. The pharmacodynamics observed in this study suggest that peak concentrations of amoxicillin of 6 to 9 microg/ml may be sufficient for the elimination of penicillin-nonsusceptible pneumococcal strains causing otitis media, especially those with intermediate resistance to amoxicillin. In vivo pharmacokinetic studies are needed to determine if these levels can be achieved in middle ear fluid with amoxicillin at 70 to 90 mg/kg/day divided into two daily doses. If these levels are reliably achieved, then clinical studies are warranted. PMID:9303386

Influence of Cooling Rate on Growth of Bacillus cereus from Spore Inocula in Cooked Rice, Beans, Pasta, and Combination Products Containing Meat or Poultry.

PubMed

Juneja, Vijay K; Mohr, Tim B; Silverman, Meryl; Snyder, O Peter

2018-02-23

The objective of this study was to assess the ability of Bacillus cereus spores to germinate and grow in order to determine a safe cooling rate for cooked rice, beans, and pasta, rice-chicken (4:1), rice-chicken-vegetables (3:1:1), rice-beef (4:1), and rice-beef-vegetables (3:1:1). Samples were inoculated with a cocktail of four strains of heat-shocked (80°C for 10 min) B. cereus spores (NCTC 11143, 935A/74, Brad 1, and Mac 1) to obtain a final spore concentration of approximately 2 log CFU/g. Thereafter, samples were exponentially cooled through the temperature range of 54.5 to 7.2°C in 6, 9, 12, 15, 18, and 21 h. At the end of the cooling period, samples were removed and plated on mannitol egg yolk polymyxin agar. The plates were incubated at 30°C for 24 h. The net B. cereus growth from spores in beans was <1 log after 9 h of cooling, but the pathogen grew faster in rice and pasta. In combination products, the net growth was as follows: 3.05, 3.89, and 4.91 log CFU/g in rice-chicken; 3.49, 4.28, and 4.96 log CFU/g in rice-beef; 3.50, 4.20, and 5.32 CFU/g in rice-chicken-mixed vegetables; and 3.68, 4.44, and 5.25 CFU/g in rice-beef-mixed vegetables after 15, 18, and 21 h of cooling, respectively. This study suggests safe cooling rates for cooling cooked rice, beans, pasta, rice-chicken, rice-chicken-vegetables, rice-beef, and rice-beef-vegetables to guard against the hazards associated with B. cereus.

Correlation of in vitro time-kill curves and kinetics of bacterial killing in cerebrospinal fluid during ceftriaxone therapy of experimental Escherichia coli meningitis.

PubMed Central

Decazes, J M; Ernst, J D; Sande, M A

1983-01-01

Ceftriaxone was highly active in eliminating Escherichia coli from the cerebrospinal fluid of rabbits infected with experimental meningitis. However, concentrations equal to or greater than 10 times the minimal bactericidal concentration had to be achieved to ensure optimal efficacy (rate of kill, 1.5 log10 CFU/ml per h). In contrast to other beta-lactams studied in this model, ceftriaxone concentrations in cerebrospinal fluid progressively increased, whereas serum steady state was obtained by constant infusion. The percent penetration was 2.1% after 1 h of therapy, in contrast to 8.9% after 7 h (P less than 0.001). In vitro time-kill curves done in cerebrospinal fluid or broth more closely predicted the drug concentrations required for a maximum cidal effect in vivo than that predicted by determinations of minimal inhibitory or bactericidal concentrations. PMID:6316841

Microbial risk assessment with the OAEL approach at water abstraction points in rural Kenya

NASA Astrophysics Data System (ADS)

Yillia, Paul T.; Kreuzinger, Norbert; Mathooko, Jude M.; Ndomahina, Ernest T.

US-based models for recreational water quality were applied to characterize the potential health risk (PHR) of infection with gastroenteritis (GI) and highly credible gastroenteritis (HCGI) illnesses from single exposure at several water abstraction points (WAPs) along the Njoro River in rural Kenya. Ambient geometric mean densities of Escherichia coli (EC) and intestinal enterococci (IE) were generally high (2-4 log units of cfu/100 ml) and risk levels were grossly in excess of acceptable health risk (AHR) levels for bathing and drinking. PHR was 2-3 times higher with the Cabelli (IE) model (Equation (2)) compared to the US EPA (EC) model (Equation (1)). Risk levels varied among WAPs in concomitance to the spatial and seasonal variability of ambient EC and IE densities. With the Cabelli IE model, PHR of HCGI illness on single exposure to the dry weather 95th percentile IE density for bathing was 2.5% of the exposed population at Logoman compared to 5.2% at Turkana Flats, 4.9% at Kenyatta or Nessuit and 4.6%, 4.5% and 4.2% at Treetop, Segotik and Njoro Bridge, respectively. PHR was ⩾5% on exposure to the wet weather 95th percentile IE density at all WAPs, excepting Treetop with 4.3%. Relative risk levels increased by at least 30 and 70 times for GI and HCGI illnesses, respectively, from drinking (250 ml) raw stream water, rising erratically in wet weather by >80% of the dry weather risk at Logoman, >30% at Njoro Bridge and Kenyatta and 10-15% at Segotik, Nessuit and Turkana Flats. By stipulating freshwater bathing water quality guidelines of 126 and 33 cfu/100 ml for EC and IE, respectively, US, EPA upholds maximum AHR levels at 0.7% and 1.9% for EC and IE, respectively. Hence, reducing current PHR levels at the WAPs to the US, EPA bathing AHR levels would require at least 2-4 log reductions of IE and EC densities with even further log reductions to achieve the WHO recommended drinking water AHR level of 0.1%. This would necessitate specialized treatment, in particular point-of-use treatment at the household level, as well as the implementation of comprehensive catchment management measures to protect the stream and the WAPs.

Comparative efficacy of Zataria multiflora Boiss., Origanum compactum and Eugenia caryophyllus essential oils against E. coli O157:H7, feline calicivirus and endogenous microbiota in commercial baby-leaf salads.

PubMed

Azizkhani, Maryam; Elizaquível, Patricia; Sánchez, Gloria; Selma, María Victoria; Aznar, Rosa

2013-09-02

Ready-to-eat salads using baby-leaf and multi-leaf mixes are one of the most promising developments in the fresh-cut food industry. There is great interest in developing novel decontamination treatments, which are both safe for consumers and more efficient against foodborne pathogens. In this study, emulsions of essential oils (EOs) from Origanum compactum (oregano), Eugenia caryophyllus (clove), and Zataria multiflora Boiss (zataria) were applied by spray (0.8 ml) after the sanitizing washing step. The aim was to investigate their ability to control the growth of potentially cross-contaminating pathogens and endogenous microbiota in commercial baby leaves, processed in a fresh-cut produce company. Zataria EO emulsions of 3%, 5% and 10% reduced Escherichia coli O157:H7 by 1.7, 2.2 and 3.5 log cfu/g in baby-leaf salads after 5 days of storage at 7°C. By contrast, reductions in E. coli O157:H7 counts remained the same when clove was applied at concentrations of 5% and 10% (2.5 log cfu/g reduction). Oregano (10%) reduced inoculated E. coli O157:H7 counts in baby-leaf salads by a maximum of 0.5 log cfu/g after 5 days of storage. Zataria showed strong antimicrobial efficacy against E. coli O157:H7 and also against the endogenous microbiota of baby-leaf salads stored for 9 days. Feline calicivirus (FCV), a norovirus surrogate, survived on inoculated baby-leaf salads during refrigerated storage (9 days at 7°C) regardless of treatment. Refrigeration temperatures completely annulled the effectiveness of the EOs against FCV inoculated in baby-leaf salads as occurred in FCV cultures. This study shows that EOs, and zataria in particular, have great potential use as an additional barrier to reduce contamination-related risks in baby-leaf salads. However, further research should be done into foodborne viruses in order to improve food safety. © 2013.

Simultaneous differential detection of human pathogenic and nonpathogenic Vibrio species using a multiplex PCR based on gyrB and pntA genes.

PubMed

Teh, C S J; Chua, K H; Thong, K L

2010-06-01

To develop a multiplex PCR targeting the gyrB and pntA genes for Vibrio species differentiation. Four pairs of primers targeting gyrB gene of Vibrios at genus level and pntA gene of Vibrio cholerae, Vibrio parahaemolyticus, Vibrio vulnificus were designed. This PCR method precisely identified 250 Vibrio species and demonstrated sensitivity in the range of 4 x 10(4) CFU ml(-1) (c. 200 CFU per PCR) to 2 x 10(3) CFU ml(-1) (c. 10 CFU per PCR). Overall, the gyrB gene marker showed a higher specificity than the dnaJ gene marker for Vibrio detection and was able to distinguish Aeromonas from Vibrio species. The multiplex PCR based on combined gyrB and pntA provides a high discriminatory power in the differentiation between Vibrio alginolyticus and V. parahaemolyticus, and between V. cholerae and Vibrio mimicus. This assay will be useful for rapid differentiation of various Vibrio species from clinical and environmental sources and significantly overcomes the limitations of the conventional methods.

Fate of Escherichia coli O157:H7 in mechanically tenderized beef prime rib following searing, cooking, and holding under commercial conditions.

PubMed

Porto-Fett, Anna C S; Shoyer, Bradley A; Thippareddi, Harshavardhan; Luchansky, John B

2013-03-01

We evaluated the effect of commercial times and temperatures for searing, cooking, and holding on the destruction of Escherichia coli O157:H7 (ECOH) within mechanically tenderized prime rib. Boneless beef ribeye was inoculated on the fat side with ca. 5.7 log CFU/g of a five-strain cocktail of ECOH and then passed once through a mechanical tenderizer with the fat side facing upward. The inoculated and tenderized prime rib was seared by broiling at 260°C for 15 min in a conventional oven and then cooked in a commercial convection oven at 121.1°C to internal temperatures of 37.8, 48.9, 60.0, and 71.1°C before being placed in a commercial holding oven maintained at 60.0°C for up to 8 h. After searing, ECOH levels decreased by ca. 1.0 log CFU/g. Following cooking to internal temperatures of 37.8 to 71.1°C, pathogen levels decreased by an additional ca. 2.7 to 4.0 log CFU/g. After cooking to 37.8, 48.9, or 60.0°C and then warm holding at 60.0°C for 2 h, pathogen levels increased by ca. 0.2 to 0.7 log CFU/g. However, for prime rib cooked to 37.8°C, pathogen levels remained relatively unchanged over the next 6 h of warm holding, whereas for those cooked to 48.9 or 60.0°C pathogen levels decreased by ca. 0.3 to 0.7 log CFU/g over the next 6 h of warm holding. In contrast, after cooking prime rib to 71.1°C and holding for up to 8 h at 60.0°C, ECOH levels decreased by an additional ca. 0.5 log CFU/g. Our results demonstrated that to achieve a 5.0-log reduction of ECOH in blade tenderized prime rib, it would be necessary to sear at 260°C for 15 min, cook prime rib to internal temperatures of 48.9, 60.0, or 71.1°C, and then hold at 60.0°C for at least 8 h.

Inactivation of Escherichia coli in a tropical fruit smoothie by a combination of heat and pulsed electric fields.

PubMed

Walkling-Ribeiro, M; Noci, F; Cronin, D A; Lyng, J G; Morgan, D J

2008-10-01

Moderate heat in combination with pulsed electric fields (PEF) was investigated as a potential alternative to thermal pasteurization of a tropical fruit smoothie based on pineapple, banana, and coconut milk, inoculated with Escherichia coli K12. The smoothie was heated from 25 degrees C to either 45 or 55 degrees C over 60 s and subsequently cooled to 10 degrees C. PEF was applied at electric field strengths of 24 and 34 kV/cm with specific energy inputs of 350, 500, and 650 kJ/L. Both processing technologies were combined using heat (45 or 55 degrees C) and the most effective set of PEF conditions. Bacterial inactivation was estimated on standard and NaCl-supplemented tryptone soy agar (TSA) to enumerate sublethally injured cells. By increasing the temperature from 45 to 55 degrees C, a higher reduction in E. coli numbers (1 compared with 1.7 log(10) colony forming units {CFU} per milliliter, P < 0.05) was achieved. Similarly, as the field strength was increased during stand-alone PEF treatment from 24 to 34 kV/cm, a greater number of E. coli cells were inactivated (2.8 compared with 4.2 log(10) CFU/mL, P < 0.05). An increase in heating temperature from 45 to 55 degrees C during a combined heat/PEF hurdle approach induced a higher inactivation (5.1 compared with 6.9 log(10) CFU/mL, respectively [P < 0.05]) with the latter value comparable to the bacterial reduction of 6.3 log(10) CFU/mL (P> or = 0.05) achieved by thermal pasteurization (72 degrees C, 15 s). A reversed hurdle processing sequence did not affect bacterial inactivation (P> or = 0.05). No differences were observed (P> or = 0.05) between the bacterial counts estimated on nonselective and selective TSA, suggesting that sublethal cell injury did not occur during single PEF treatments or combined heat/PEF treatments.

Survival of Salmonella, Escherichia coli O157:H7, and Listeria monocytogenes on raw peanut and pecan kernels stored at -24, 4, and 22°C.

PubMed

Brar, Pardeepinder K; Proano, Lisseth G; Friedrich, Loretta M; Harris, Linda J; Danyluk, Michelle D

2015-02-01

Cocktails of lawn-collected cells were used to determine the survival of Salmonella, Escherichia coli O157:H7, and Listeria monocytogenes on the surface of raw peanut and pecan kernels. Kernels were inoculated with mixtures of four to five strains at 3 or 6 log CFU/g, dried at room temperature, and then stored at -24 ± 1, 4 ± 2, and 22 ± 1°C for 28 or 365 days. In most cases, rates of decline of the pathogens did not differ significantly between the two inoculum concentrations in the 28-day study. At 6 log CFU/g, populations of all pathogens were reduced by 0.5 to 1.6 log CFU/g during an initial 3-day drying period on both peanuts and pecans. The moisture content of peanuts and pecans remained stable at -24 ± 1 and 22 ± 1°C; at 4 ± 2°C, the moisture content increased from 3.8 to 5.6% on peanuts and from 2.6 to 3% on pecans over 365 days. Pathogen populations were stable on pecans stored under frozen and refrigerated conditions, except for L. monocytogenes, which declined at a rate of 0.03 log CFU/g/30 days at 4 ± 2°C. Salmonella populations were stable on peanuts stored at -24 ± 1 and 4 ± 2°C, but E. coli O157:H7 and L. monocytogenes declined at rates of 0.03 to 0.12 log CFU/g/30 days. At 22 ± 1°C, Salmonella, E. coli O157:H7, and L. monocytogenes declined at a rate of 0.22, 0.37, and 0.59 log CFU/g/30 days, respectively, on peanuts, and at 0.15, 0.34, and 1.17 log CFU/g/30 days, respectively, on pecans. Salmonella counts were above the limit of detection (0.30 log CFU/g) throughout the study. In most cases during storage, counts obtained from pecans were higher than from peanuts.

Post-processing application of chemical solutions for control of Listeria monocytogenes, cultured under different conditions, on commercial smoked sausage formulated with and without potassium lactate-sodium diacetate.

PubMed

Geornaras, Ifigenia; Skandamis, Panagiotis N; Belk, Keith E; Scanga, John A; Kendall, Patricia A; Smith, Gary C; Sofos, John N

2006-12-01

This study evaluated post-processing chemical solutions for their antilisterial effects on commercial smoked sausage formulated with or without 1.5% potassium lactate plus 0.05% sodium diacetate, and contaminated (approximately 3-4 log cfu/cm(2)) with 10-strain composite Listeria monocytogenes inocula prepared under various conditions. Inoculated samples were left untreated, or were immersed (2 min, 25 +/- 2 degrees C) in solutions of acetic acid (2.5%), lactic acid (2.5%), potassium benzoate (5%) or Nisaplin (0.5%, equivalent to 5000 IU/ml of nisin) alone, and in sequence (Nisaplin followed by acetic acid, lactic acid or potassium benzoate), before vacuum packaging and storage at 10 degrees C (48 days). Acetic acid, lactic acid or potassium benzoate applied alone reduced initial L. monocytogenes populations by 0.4-1.5 log cfu/cm(2), while treatments including Nisaplin caused reductions of 2.1-3.3 log cfu/cm(2). L. monocytogenes on untreated sausage formulated with antimicrobials had a lag phase duration of 10.2 days and maximum specific growth rate (mu(max)) of 0.089 per day, compared to no lag phase and mu(max) of 0.300 per day for L. monocytogenes on untreated product that did not contain antimicrobials in the formulation. The immersion treatments inhibited growth of the pathogen for 4.9-14.8 days on sausage formulated without potassium lactate-sodium diacetate; however, in all cases significant (P < 0.05) growth occurred by the end of storage. The antilisterial activity of chemical solutions was greatly enhanced when applied to product formulated with antimicrobials; growth was completely inhibited on sausage treated with acetic or lactic acid alone, and in sequence with Nisaplin. In general, habituation (15 degrees C, 7 days) of L. monocytogenes cells, planktonically or as attached cells to stainless-steel coupons in sausage homogenate prior to contamination of product, resulted in shorter lag phase durations compared with cells cultivated planktonically in a broth medium. Furthermore, when present, high levels of spoilage flora were found to suppress growth of the pathogen. Findings of this study could be useful to US meat processors in their efforts to select required regulatory alternatives for control of post-processing contamination in meat products.

Borosilicate Glass Fiber-Optic Biosensor for the Detection of Escherichia coli.

PubMed

Maas, Michael B; Maybery, Giles H C; Perold, Willem J; Neveling, Deon P; Dicks, Leon M T

2018-02-01

Polyclonal antibodies against Escherichia coli and fluorescent, secondary, antibodies were immobilized on borosilicate glass fibers pre-treated with 3-glycidyloxypropyl trimethoxysilane (GPS). Light with an average wavelength of 627 nm, emitted by a diode placed at one end of the glass fiber, was detected by an ultrasensitive photodiode with peak sensitivity at 640 nm. Changes in fluorescence, caused by binding of E. coli to the antibodies, changed the net refractive index of the glass fiber and thus the internal reflection of light. These evanescent changes in photon energy were recorded by an ultrasensitive photodiode. Signals were amplified and changes in voltage recorded with a digital multimeter. A linear increase in voltage readings was recorded over 2 h when 3.0 × 10 7 CFU/ml and 2.77 × 10 9 CFU/ml E. coli were adhered to the antibodies. Voltage readings were recorded with E. coli cell numbers from 2 × 10 3 CFU/ml to 2 × 10 6 CFU/ml, but readings remained unchanged for 2 h, indicating that the limit of detection is 3.0 × 10 7 CFU/ml. This simple technology may be used to develop a low-cost, portable, fiber-optic biosensor to detect E. coli in infections and may have applications in the medical field. Research is in progress to optimize the sensitivity of the fiber-optic biosensor and determine its specificity.

Hyperbaric oxygen therapy augments the photodynamic action of methylene blue against bacteria in vitro

NASA Astrophysics Data System (ADS)

Bisland, S. K.; Dadani, F. N.; Chien, C.; Wilson, B. C.

2007-02-01

Photodynamic therapy (PDT) entails the combination of photosensitizer and light to generate cytotoxic molecules that derive from molecular oxygen (O II). The presence of sufficient O II within the target tissues is critical to the efficiency of PDT. This study investigates the use of hyperbaric oxygen therapy in combination with PDT (HOTPDT) to augment the photodynamic action of methylene blue (MB) or 5-aminolevulinic acid (ALA) against gram positive and gram negative bacterial strains in vitro. Staphylococcus aureus or Pseudomonas aeruginosa were grown in trypticase soy broth as planktonic cultures (~10 8/mL) or as established biofilms in 48 well plates (3 days old) at 32°C. Dark toxicity and PDT response in the presence or absence of HOT (2 atmospheres, 100% O II for 30, 60 or 120 min) was established for both MB (0-0.1 mM) and ALA (0- 1 mM) for a range of incubation times. The number of surviving colonies (CFU/mL) was plotted for each treatment groups. Light treatments (5, 10, 20 or 30 J/cm2) were conducted using an array of halogen bulbs with a red filter providing 90% transmittance over 600-800 nm at 21 mW/cm2. HOT increased the dark toxicity of MB (30 min, 0.1 mM) from < 0.2 log cell kill to 0.5 log cell kill. Dark toxicity of ALA (4 hr, 1 mM) was negligible and did not increase with HOT. For non-dark toxic concentrations of MB or ALA, (0.05 mM and 1 mM respectively) HOT-PDT enhanced the antimicrobial effect of MB against Staphylococcus aureus in culture by >1 and >2 logs of cell kill (CFU/mL) at 5 and 10 J/cm2 light dose respectively as compared to PDT alone. HOT-PDT also increased the anti-microbial effects of MB against Staphylococcus aureus biofilms compared to PDT, albeit less so (> 2 logs) following 10 J/cm2 light dose. Anti-microbial effects of PDT using ALA were not significant for either strain with or without HOT. These data suggest that HOTPDT may be useful for improving the PDT treatment of bacterial infections.

Evaluation of modified dichloran 18% glycerol (DG18) agar for enumerating fungi in wheat flour: a collaborative study.

PubMed

Beuchat, L R; Hwang, C A

1996-04-01

Dichloran 18% glycerol agar base supplemented with 100 micrograms of chloramphenicol ml-1 (DG18 agar) was compared to DG18 agar supplemented with 100 micrograms of Triton X-301 ml-1 (DG18T) and DG18 agar supplemented with 1 microgram of iprodione [3-(3,5-dichlorophenyl)-N-(1-methyl-ethyl)-2,4-dioxo-1-imidazolidine- carboxamide] ml-1 (DG18I agar) for enumeration of fungi in ten brands of wheat flour. As the flours contained low fungal populations, all were inoculated with two to four strains of xerophilic fungi (Aspergillus candidus, A. penicillioides, Eurotium amstelodami, E. intermedium, E. repens, E. rubrum, E. tonophilum, E. umbrosum and Wallemia sebi), after which counts ranged from 3.87 to 6.37 log10 CFU g-1. Significantly higher populations (p < 0.05) were detected in four flours: three were on DG18T compared to DG18 and DG18I agar. A. candidus had been inoculated into all three flours. E. amstelodami, E. intermedium, E. repens or E. tonophilum had also been inoculated into at least one of the three flours showing significantly higher numbers of CFU on DG18T agar. Analysis of collapsed data from all samples showed that DG18T agar was significantly better than DG18 or DG18I agars at p < 0.10 but not at p < 0.05. Coefficients of variation for reproducibility (among-laboratory variation) were 8.4%, 7.5% and 8.6%, respectively, for DG18, DG18T and DG18I agars. DG18I agar restricted colony development most, especially for Eurotium species. Naturally occurring Penicillium species grew equally well on DG18 and DG18T agars, whereas W. sebi grew well on all three media. DG18T agar was judged to be superior to DG18 and DG18I agars for enumerating fungi in wheat flours.

An in vitro evaluation of the antibacterial efficacy of chlorine dioxide on E. faecalis in bovine incisors.

PubMed

Eddy, Russell S; Joyce, Anthony P; Roberts, Steven; Buxton, Thomas B; Liewehr, Frederick

2005-09-01

This study investigated the ability of chlorine dioxide to eliminate Enterococcus faecalis from dentinal tubules of bovine incisors. Thirty-seven extracted bovine incisor roots were sectioned into seventy-four 5 mm disks. Standardized lumens were filled with either sterile Brain Heart Infusion Broth (contamination controls, n = 10) or BHI containing E. faecalis (1.0 x 10 cfu/ml). Disks were incubated in 5% CO2 at 37 degrees C for 72 h. To simulate endodontic instrumentation the lumens were again enlarged. Sixty disks were randomly divided into four experimental groups and filled with one of the following irrigants: 10% Clidox-S (chlorine dioxide), 13.8% BioClenz (chlorine dioxide), 5.25% Clorox, or saline. The disks were incubated for 30 min and were then frozen, pulverized, serially diluted in phosphate buffered saline, and plated on BHI plates in triplicate. Total colony forming units were counted macroscopically. Statistical analysis of the data was performed with a Kruskal-Wallis one-way ANOVA on ranks (p < 0.05, n = 60). Bacterial counts, expressed in log10 cfu/disk were as follows (">" denotes significant differences): Saline > Clidox-S = BioClenz > Clorox. All negative controls were sterile. Chlorine dioxide and NaOCL were both effective in eliminating E. faecalis from the dentinal disks within 30 min.

Application of a novel antimicrobial coating on roast beef for inactivation and inhibition of Listeria monocytogenes during storage.

PubMed

Wang, Luxin; Zhao, Liang; Yuan, Jing; Jin, Tony Z

2015-10-15

The antilisterial efficacy of novel coating solutions made with organic acids, lauric arginate ester, and chitosan was evaluated in a three-stage study on inoculated roast beef for the first time. Ready-to-eat roast beef was specially ordered from the manufacturer. The meat surface was inoculated with five-strain Listeria monocytogenes cocktail inoculums at two different levels, ~3 and 6 Log CFU/cm(2) and treated with the stock solution (HAMS), the 1:5 diluted solution (MAMS), and the 1:10 diluted solution (LAMS) (stage 1). During the 20 min contact time, the antimicrobial coatings reduced the Listeria populations by approximately 0.9-0.3 Log CFU/cm(2). The higher the concentrations of the antimicrobial solution, the better the antilisterial effects were. The treated inoculated beef samples were then stored at 4 °C for 30 days. During storage, Listeria growth inhibition effects were seen. While no growth was seen from the HAMS-treated samples, a 1.6 Log CFU/cm(2) increase was seen for MAMS-treated samples, a 4.6 Log CFU/cm(2) increase was seen for LAMS-treated samples, and a 5.7 Log CFU/cm(2) increase was seen for NoAMS-treated samples on Day 30 (~3 Log CFU/cm(2) inoculation level). In the second stage, the impact of the roast beef storage time on solution's antilisterial effect was evaluated. Results showed that the effect of the antimicrobial solution was dependent on both the initial inoculation levels and storage times. In stage 3, the effect of the antimicrobial solution on roast beef quality was studied with both instrument measurement and sensory evaluation. Minor changes in color, pH, and water activity were found. However, only limited sensory differences were seen between the treated and untreated samples. When panels were able to accurately find color differences between samples, they preferred the treated samples. The findings of this research proved the antilisterial efficacy of the novel antimicrobial solution and showed its potential for being used as a roast beef cut surface coating to control Listeria contamination and for color protection. Copyright © 2015 Elsevier B.V. All rights reserved.

Effect of storage temperatures and stresses on the survival of Salmonella spp. in halva.

PubMed

Osaili, T M; Al-Nabulsi, A A; Nazzal, D S; Shaker, R R

2017-11-01

The presence of Salmonella spp. in halva has been associated with foodborne illnesses and product recalls from the markets. This study investigated the effect of environmental stresses on the survival of Salmonella spp. in halva during storage for 12 months at 10 and 25°C (log (N 0 /N) g -1 ). Halva samples were inoculated with a cocktail of four strains of unstressed, desiccation stressed or heat stressed Salmonella (10 6 -10 7  CFU per gram). In general, survival of Salmonella spp. in halva decreased significantly (P ˂ 0·05) as storage time and temperature increased. At the end of halva shelf life at 10°C, the initial populations of unstressed, desiccation stressed or heat stressed Salmonella spp. decreased by 2·7, 2·6 or 2·8 log CFU per gram (reduction rate c. 0·2 log CFU per month), respectively. While at 25°C, the populations decreased 5·2, 6·7 or 6·3 log CFU per gram, respectively (reduction rate c. 0·4-0·5 log CFU per month). The populations of stressed Salmonella spp. in halva samples were not significantly different (P ≥ 0·05) from populations of unstressed cells during storage at 10 and 25°C, except during the last 3 months of storage at 25°C when populations of unstressed cells were higher (P < 0·05). Exposing Salmonella spp. to desiccation or heat stress prior product contamination may play a role in Salmonella spp. survival in halva during storage. Contamination of halva (tahini halva) with Salmonella from raw materials or during production was documented. Halva and tahini have been involved in salmonellosis outbreaks in different countries. The study demonstrated enhanced survivability of stressed and unstressed Salmonella spp. in halva over a 12-month storage period at 10 and 25°C with lower log reductions than expected. Exposing Salmonella spp. to desiccation or heat stress prior product contamination may play a role in microbial survival in halva during storage. These findings serve as a model to halva producers to implement control measures to prevent Salmonella spp. contamination in halva. © 2017 The Society for Applied Microbiology.

Phage Inactivation of Listeria monocytogenes on San Daniele Dry-Cured Ham and Elimination of Biofilms from Equipment and Working Environments

PubMed Central

Iacumin, Lucilla; Manzano, Marisa; Comi, Giuseppe

2016-01-01

The anti-listerial activity of generally recognized as safe (GRAS) bacteriophage Listex P100 (phage P100) was demonstrated in broths and on the surface of slices of dry-cured ham against 5 strains or serotypes (i.e., Scott A, 1/2a, 1/2b, and 4b) of Listeria monocytogenes. In a broth model system, phage P100 at a concentration equal to or greater than 7 log PFU/mL completely inhibited 2 log CFU/cm2 or 3 log CFU/cm2 of L. monocytogenes growth at 30 °C. The temperature (4, 10, 20 °C) seemed to influence P100 activity; the best results were obtained at 4 °C. On dry-cured ham slices, a P100 concentration ranging from 5 to 8 log PFU/cm2 was required to obtain a significant reduction in L. monocytogenes. At 4, 10, and 20 °C, an inoculum of 8 log PFU/cm2 was required to completely eliminate 2 log L. monocytogenes/cm2 and to reach the absence in 25 g product according to USA food law. Conversely, it was impossible to completely eradicate L. monocytogenes with an inoculum of approximately of 3.0 and 4.0 log CFU/cm2 and with a P100 inoculum ranging from 1 to 7 log PFU/cm2. P100 remained stable on dry-cured ham slices over a 14-day storage period, with only a marginal loss of 0.2 log PFU/cm2 from an initial phage treatment of approximately 8 log PFU/cm2. Moreover, phage P100 eliminated free L. monocytogenes cells and biofilms on the machinery surfaces used for dry-cured ham production. These findings demonstrate that the GRAS bacteriophage Listex P100 at level of 8 log PFU/cm2 is listericidal and useful for reducing the L. monocytogenes concentration or eradicating the bacteria from dry-cured ham. PMID:27681898

[In vitro susceptibilities of causative organisms isolated from patients with primary respiratory tract infections to BRL 25000 (clavulanic acid/amoxicillin)].

PubMed

Deguchi, K; Fukayama, S; Nishimura, Y; Yokota, N; Tanaka, S; Oda, S; Matsumoto, Y; Ikegami, R; Sato, K; Fukumoto, T

1985-10-01

The in vitro susceptibilities of various causative organisms recently isolated from patients with primary respiratory tract infections to BRL 25000 (a formulation of amoxicillin, 2 parts, and potassium clavulanate, 1 part), amoxicillin (AMPC), cefaclor (CCL), cephalexin (CEX), cefadroxil (CDX) and cefroxadine (CXD) were determined. beta-Lactamase producing strains were detected by nitrocefin chromogenic method and PCG acidometric method. The frequency of isolation of beta-lactamase production in strains of S. aureus, H. influenzae, B. catarrhalis and K. pneumoniae was 92%, 18%, 36% and 98%, respectively. Against S. aureus strains with MIC values to AMPC of less than or equal to 100 micrograms/ml and CEX of less than or equal to 25 micrograms/ml BRL 25000 showed MIC values in the range 0.39-6.25 micrograms/ml with inocula of 10(6) CFU/ml, while BRL 25000 required 12.5-100 micrograms/ml of concentrations for inhibition of the strains with MIC values to AMPC of greater than 100 micrograms/ml and CEX of greater than or equal to 25 micrograms/ml. Against S. pyogenes and S. pneumoniae BRL 25000 showed MIC values in the range less than 0.024-0.10 micrograms/ml with inocula of 10(6) CFU/ml, which is much more active than CCL, CEX, CDX and CXD and slight less active than AMPC. Against H. influenzae and B. catarrhalis BRL 25000 showed MIC values in the range 0.20-6.25 micrograms/ml with inocula of 10(6) CFU/ml, which showed most potent activity among the agents tested. The activity of BRL 25000 against K. pneumoniae was approximately equal to that of CCL and superior to that of AMPC, CEX, CDX and CXD.

Identification of the cell binding domain in Nipah virus G glycoprotein using a phage display system.

PubMed

Lam, Chui-Wan; AbuBakar, Sazaly; Chang, Li-Yen

2017-05-01

Nipah virus (NiV) is a highly pathogenic zoonotic paramyxovirus with unusual broad host tropism and is designated as a Category C pathogen by the U.S. National Institute of Allergy and Infectious Diseases. NiV infection is initiated after binding of the viral G glycoprotein to the host cell receptor. The aim of this study was to map the NiV G glycoprotein cell binding domain using a phage display system. The NiV G extracellular domain was truncated and displayed as attachment proteins on M13 phage g3p minor coat protein. The binding efficiency of recombinant phages displaying different regions of NiV G to mammalian cells was evaluated. Results showed that regions of NiV G consisting of amino acids 396-602 (recombinant phage G4) and 498-602 (recombinant phage G5) demonstrated the highest binding to both Vero (5.5×10 3 cfu/ml and 5.6×10 3 cfu/ml) and THP-1 cells (3.5×10 3 cfu/ml and 2.9×10 3 cfu/ml). However, the binding of both of these recombinant phages to THP-1 cells was significantly lower than to Vero cells, and this could be due to the lack of primary host cell receptor expression on THP-1 cells. Furthermore, the binding between these two recombinant phages was competitive suggesting that there was a common host cell attachment site. This study employed an approach that is suitable for use in a biosafety level 2 containment laboratory without the need to use live virus to show that NiV G amino acids 498-602 play an important role for attachment to host cells. Copyright © 2017 Elsevier B.V. All rights reserved.

Control of Salmonella on sprouting mung bean and alfalfa seeds by using a biocontrol preparation based on antagonistic bacteria and lytic bacteriophages.

PubMed

Ye, Jianxiong; Kostrzynska, Magdalaena; Dunfield, Kari; Warriner, Keith

2010-01-01

The following reports on the application of a combination of antagonistic bacteria and lytic bacteriophages to control the growth of Salmonella on sprouting mung beans and alfalfa seeds. Antagonistic bacteria were isolated from mung bean sprouts and tomatoes by using the deferred plate assay to assess anti-Salmonella activity. From the isolates screened, an Enterobacter asburiae strain (labeled "JX1") exhibited stable antagonistic activity against a broad range of Salmonella serovars (Agona, Berta, Enteritidis, Hadar, Heidelberg, Javiana, Montevideo, Muenchen, Newport, Saint Paul, and Typhimurium). Lytic bacteriophages against Salmonella were isolated from pig or cattle manure effluent. A bacteriophage cocktail prepared from six isolates was coinoculated with E. asburiae JX1 along with Salmonella in broth culture. The combination of E. asburiae JX1 and bacteriophage cocktail reduced the levels of Salmonella by 5.7 to 6.4 log CFU/ml. Mung beans inoculated with Salmonella and sprouted over a 4-day period attained levels of 6.72 + or - 0.78 log CFU/g. In contrast, levels of Salmonella were reduced to 3.31 + or - 2.48 or 1.16 + or - 2.14 log CFU/g when the pathogen was coinoculated with bacteriophages or E. asburiae JX1, respectively. However, by using a combination of E. asburiae JX1 and bacteriophages, the levels of Salmonella associated with mung bean sprouts were only detected by enrichment. The biocontrol preparation was effective at controlling the growth of Salmonella under a range of sprouting temperatures (20 to 30 degrees Celsius) and was equally effective at suppressing the growth of Salmonella on sprouting alfalfa seeds. The combination of E. asburiae JX1 and bacteriophages represents a promising, chemical-free approach for controlling the growth of Salmonella on sprouting seeds.

Analysis of alcohol-based hand sanitizer delivery systems: efficacy of foam, gel, and wipes against influenza A (H1N1) virus on hands.

PubMed

Larson, Elaine L; Cohen, Bevin; Baxter, Kathleen A

2012-11-01

Minimal research has been published evaluating the effectiveness of hand hygiene delivery systems (ie, rubs, foams, or wipes) at removing viruses from hands. The purposes of this study were to determine the effect of several alcohol-based hand sanitizers in removing influenza A (H1N1) virus, and to compare the effectiveness of foam, gel, and hand wipe products. Hands of 30 volunteers were inoculated with H1N1 and randomized to treatment with foam, gel, or hand wipe applied to half of each volunteer's finger pads. The log(10) count of each subject's treated and untreated finger pads were averaged. Log(10) reductions were calculated from these differences and averaged within treatment group. Between-treatment analysis compared changes from the untreated finger pads using analysis of covariance with treatment as a factor and the average log(10) untreated finger pads as the covariate. Log(10) counts on control finger pads were 2.7-5.3 log(10) of the 50% infectious dose for tissue culture (TCID(50)/0.1 mL) (mean, 3.8 ± 0.5 log(10) TCID(50)/0.1 mL), and treated finger pad counts for all test products were 0.5-1.9 log(10) TCID(50)/0.1 mL (mean, 0.53 ± 0.17 log(10) TCID(50)/0.1 mL). Treatments with all products resulted in a significant reduction in viral titers (>3 logs) at their respective exposure times that were statistically comparable. All 3 delivery systems (foam, gel, and wipe) produced significantly reduced viral counts on hands. Copyright © 2012 Association for Professionals in Infection Control and Epidemiology, Inc. Published by Mosby, Inc. All rights reserved.

Influence of probiotics, included in peanut butter, on the fate of selected Salmonella and Listeria strains under simulated gastrointestinal conditions.

PubMed

Klu, Y A K; Chen, J

2016-04-01

This study observed the behaviour of probiotics and selected bacterial pathogens co-inoculated into peanut butter during gastrointestinal simulation. Peanut butter homogenates co-inoculated with Salmonella/Listeria strains (5 log CFU ml(-1) ) and lyophilized or cultured probiotics (9 log CFU ml(-1) ) were exposed to simulated gastrointestinal conditions for 24 h at 37°C. Sample pH, titratable acidity and pathogen populations were determined. Agar diffusion assay was performed to assess the inhibitory effect of probiotic culture supernatants with either natural (3·80 (Lactobacillus), 3·78 (Bifidobacteirum) and 5·17 (Streptococcus/Lactococcus)) or neutralized (6·0) pH. Antibacterial effect of crude bacteriocin extracts were also evaluated against the pathogens. After 24 h, samples with probiotics had lower pH and higher titratable acidity than those without probiotics. The presence of probiotics caused a significant reduction (P < 0·05) in pathogen populations. Supernatants of Bifidobacterium and Lactobacillus cultures inhibited pathogen growth; however, the elevation of pH diminished their antibacterial activities. Crude bacteriocin extracts had a strain-specific inhibitory effect only towards Listeria monocytogenes. Probiotics in 'peanut butter' survived simulated gastrointestinal conditions and inhibited the growth of Salmonella/Listeria. Peanut butter is a plausible carrier to deliver probiotics to improve the gastrointestinal health of children in developing countries. © 2016 The Society for Applied Microbiology.

Growth and inactivation of Salmonella enterica and Listeria monocytogenes in broth and validation in ground pork meat during simulated home storage abusive temperature and home pan-frying

PubMed Central

Wang, Xiang; Lahou, Evy; De Boeck, Elien; Devlieghere, Frank; Geeraerd, Annemie; Uyttendaele, Mieke

2015-01-01

Ground pork meat with natural microbiota and inoculated with low initial densities (1–10 or 10–100 CFU/g) of Salmonella enterica or Listeria monocytogenes was stored under abusive temperature at 10°C and thermally treated by a simulated home pan-frying procedure. The growth and inactivation characteristics were also evaluated in broth. In ground pork meat, the population of S. enterica increased by less than one log after 12-days of storage at 10°C, whereas L. monocytogenes increased by 2.3 to 2.8 log units. No unusual intrinsic heat resistance of the pathogens was noted when tested in broth at 60°C although shoulders were observed on the inactivation curves of L. monocytogenes. After growth of S. enterica and L. monocytogenes at 10°C for 5 days to levels of 1.95 log CFU/g and 3.10 log CFU/g, respectively, in ground pork meat, their inactivation in the burger subjected to a simulated home pan-frying was studied. After thermal treatment S. enterica was undetectable but L. monocytogenes was recovered in three out of six of the 25 g burger samples. Overall, the present study shows that data on growth and inactivation of broths are indicative but may underestimate as well as overestimate behavior of pathogens and thus need confirmation in food matrix conditions to assess food safety in reasonably foreseen abusive conditions of storage and usual home pan-frying of meat burgers in Belgium. PMID:26579079

Comparison of corneal collagen cross-linking (PACK-CXL) and voriconazole treatments in experimental fungal keratitis.

PubMed

Özdemir, Hüseyin Baran; Kalkancı, Ayşe; Bilgihan, Kamil; Göçün, Pınar Uyar; Öğüt, Betül; Karakurt, Funda; Erdoğan, Merve

2018-06-04

To compare the antifungal efficacy of corneal collagen cross-linking with photoactivated riboflavin (PACK-CXL) and voriconazole in experimental Fusarium solani and Candida albicans keratitis models. Sixty-four corneas of 32 New Zealand rabbits were included and divided into two main groups. Intrastromal injection of Fusarium and Candida suspensions was performed, and it was observed that keratitis was formed on the third day. Both groups were randomly separated into the following four groups: control, PACK-CXL, voriconazole and PACK-CXL combined with voriconazole. PACK-CXL was applied using 0.25% riboflavin in an accelerated Dresden protocol (total ultraviolet A dose 5.4 J/cm²). Voriconazole was applied topically as 7x1/day with a dose of 1% (10 mg/ml). Corneal buttons were excised on the tenth day, and microbiological and pathological examinations were performed. The PACK-CXL and PACK-CXL combined with voriconazole groups each had 100 colony-forming unit (CFU/ml) of reproduced micro-organisms compared with 500 CFU/ml in the voriconazole group and 1500 CFU/ml in the control group (p < 0.001) in the Fusarium keratitis model. The PACK-CXL combined with voriconazole group had 100 CFU/ml, the PACK-CXL group had 150 CFU/ml, and the voriconazole group had 200 CFU/ml of reproduced micro-organisms compared with 4000 CFU/ml in the control group (p < 0.002) in the Candida keratitis model. (p < 0.001). Fewer hyphae and non-specific stromal changes were observed in the pathological cross sections examined in subgroups that used CXL. There was less fungus reproduction and a lower keratitis score for Fusarium solani and Candida albicans in the treatment groups compared to the control groups, especially in groups that used PACK-CXL. These results suggest that it is useful to combine PACK-CXL treatment with medical treatment in the fungal keratitis algorithm at the early stage of the disease. © 2018 Acta Ophthalmologica Scandinavica Foundation. Published by John Wiley & Sons Ltd.

Comparative evaluation of calcium hypochlorite and sodium hypochlorite associated with passive ultrasonic irrigation on antimicrobial activity of a root canal system infected with Enterococcus faecalis: an in vitro study.

PubMed

de Almeida, Ana Paula; Souza, Matheus Albino; Miyagaki, Daniela Cristina; Dal Bello, Yuri; Cecchin, Doglas; Farina, Ana Paula

2014-12-01

The purpose of this study was to compare in vitro the effectiveness of calcium hypochlorite (Ca[OCl]2) and sodium hypochlorite (NaOCl) associated with passive ultrasonic irrigation in root canals of bovine teeth infected with Enterococcus faecalis. The root canals of 60 single-rooted bovine extracted teeth were enlarged up to a file 45, autoclaved, inoculated with Enterococcus faecalis, and incubated for 30 days. The samples were divided into 6 groups (n = 10) according to the protocol for decontamination: G1: no treatment; G2: distilled water; G3: 2.5% NaOCl; G4: 2.5% Ca(OCl)2; G5: 2.5% NaOCl with ultrasonic activation; and G6: 2.5% Ca(OCl)2 with ultrasonic activation (US). Microbiological testing (colony-forming unit [CFU] counting) was performed to evaluate and show, respectively, the effectiveness of the proposed treatments. Data were subjected to 1-way analysis of variance followed by the post hoc Tukey test (α = 0.05). Groups 1 and 2 showed the highest mean contamination (3.26 log10 CFU/mL and 2.69 log10 CFU/mL, respectively), which was statistically different from all other groups (P < .05). Group 6 (Ca[OCl]2 + US) showed the lowest mean contamination (1.00 log10 CFU/mL), with no statistically significant difference found in groups 3 (NaOCl), 4 (Ca[OCl]2), and 5 (NaOCl + US) (P < .05). Ca(OCl)2 as well as passive ultrasonic irrigation can aid in chemomechanical preparation, contributing in a significant way to the reduction of microbial content during root canal treatment. Copyright © 2014 American Association of Endodontists. Published by Elsevier Inc. All rights reserved.

Presence and Persistence of Salmonella enterica Serotype Typhimurium in the Phyllosphere and Rhizosphere of Spray-Irrigated Parsley

PubMed Central

Kisluk, Guy

2012-01-01

Salmonella enterica is one of the major food-borne pathogens associated with ready-to-eat fresh foods. Although polluted water might be a significant source of contamination in the field, factors that influence the transfer of Salmonella from water to the crops are not well understood, especially under conditions of low pathogen levels in water. The aim of this study was to investigate the short- and long-term (1 h to 28 days) persistence of Salmonella enterica serotype Typhimurium in the phyllosphere and the rhizosphere of parsley following spray irrigation with contaminated water. Plate counting and quantitative real-time PCR (qRT-PCR)-based methods were implemented for the quantification. By applying qRT-PCR with enrichment, we were able to show that even irrigation with water containing as little as ∼300 CFU/ml resulted in the persistence of S. Typhimurium on the plants for 48 h. Irrigation with water containing 8.5 log CFU/ml resulted in persistence of the bacteria in the phyllosphere and the rhizosphere for at least 4 weeks, but the population steadily declined with a major reduction in bacterial counts, of ∼2 log CFU/g, during the first 2 days. Higher levels of Salmonella were detected in the phyllosphere when plants were irrigated during the night compared to irrigation during the morning and during winter compared to the other seasons. Further elucidation of the mechanisms underlying the transfer of Salmonella from contaminated water to crops, as well as its persistence over time, will enable the implementation of effective irrigation and control strategies. PMID:22447598

Presence and persistence of Salmonella enterica serotype typhimurium in the phyllosphere and rhizosphere of spray-irrigated parsley.

PubMed

Kisluk, Guy; Yaron, Sima

2012-06-01

Salmonella enterica is one of the major food-borne pathogens associated with ready-to-eat fresh foods. Although polluted water might be a significant source of contamination in the field, factors that influence the transfer of Salmonella from water to the crops are not well understood, especially under conditions of low pathogen levels in water. The aim of this study was to investigate the short- and long-term (1 h to 28 days) persistence of Salmonella enterica serotype Typhimurium in the phyllosphere and the rhizosphere of parsley following spray irrigation with contaminated water. Plate counting and quantitative real-time PCR (qRT-PCR)-based methods were implemented for the quantification. By applying qRT-PCR with enrichment, we were able to show that even irrigation with water containing as little as ∼300 CFU/ml resulted in the persistence of S. Typhimurium on the plants for 48 h. Irrigation with water containing 8.5 log CFU/ml resulted in persistence of the bacteria in the phyllosphere and the rhizosphere for at least 4 weeks, but the population steadily declined with a major reduction in bacterial counts, of ∼2 log CFU/g, during the first 2 days. Higher levels of Salmonella were detected in the phyllosphere when plants were irrigated during the night compared to irrigation during the morning and during winter compared to the other seasons. Further elucidation of the mechanisms underlying the transfer of Salmonella from contaminated water to crops, as well as its persistence over time, will enable the implementation of effective irrigation and control strategies.

A 10-day vacancy period after cleaning and disinfection has no effect on the bacterial load in pig nursery units.

PubMed

Luyckx, K; Millet, S; Van Weyenberg, S; Herman, L; Heyndrickx, M; Dewulf, J; De Reu, K

2016-10-19

Biosecurity measures such as cleaning, disinfection and a vacancy period between production cycles on pig farms are essential to prevent disease outbreaks. No studies have tested the effect of a longer vacancy period on bacterial load in nursery units. The present study evaluated the effect of a 10-day vacancy period in pig nursery units on total aerobic flora, Enterococcus spp., Escherichia coli, faecal coliforms and methicillin resistant Staphylococcus aureus (MRSA). Three vacancy periods of 10 days were monitored, each time applied in 3 units. The microbiological load was measured before disinfection and at 1, 4, 7 and 10 days after disinfection. No significant decrease or increase in E. coli, faecal coliforms, MRSA and Enterococcus spp. was noticed. Total aerobic flora counts were the lowest on day 4 after disinfection (i.e. 4.07 log CFU/625 cm 2 ) (P < 0.05), but the difference with other sampling moments was limited (i.e. 0.6 log CFU/625 cm 2 ) and therefore negligible. Furthermore, this observation on day 4 was not confirmed for the other microbiological parameters. After disinfection, drinking nipples were still mostly contaminated with total aerobic flora (i.e. 5.32 log CFU/625 cm 2 ) and Enterococcus spp. (i.e. 95 % of the samples were positive) (P < 0.01); the feeding troughs were the cleanest location (total aerobic flora: 3.53 log CFU/625 cm 2 and Enterococcus spp.: 50 % positive samples) (P < 0.01). This study indicates that prolonging the vacancy period in nursery units to 10 days after disinfection with no extra biosecurity measures has no impact on the environmental load of total aerobic flora, E. coli, faecal coliforms, MRSA and Enterococcus spp..

Fate of Listeria monocytogenes on Fresh Apples under Different Storage Temperatures.

PubMed

Sheng, Lina; Edwards, Katheryn; Tsai, Hsieh-Chin; Hanrahan, Ines; Zhu, Mei-Jun

2017-01-01

Fresh apples are typically stored for up to 1 year commercially; different apple varieties require different storage temperatures to maintain their quality characteristics. There is sparse information available about Listeria monocytogenes survival on fresh apples under various storage temperatures. The objective of this study was to comprehensively evaluate the effect of storage temperature on apple fruit decay and L. monocytogenes survival. Unwaxed apple fruits of selected varieties (Fuji and Granny Smith) were dip inoculated in a three-strain L. monocytogenes cocktail to establish ∼3.5 and 6.0 Log 10 CFU/apple. Twenty-four hours post-inoculation, apples were subjected to 1, 4, 10, or 22°C storage for up to 3 months. Apples under the different storage treatments were sampled at 1-, 4-, 7- and 14-day for short-term storage under all four tested temperatures, and 2-, 4-, 8-, and 12-week for long-term storage at 1, 4, and 10°C. A set of uninoculated and unwaxed apples were simultaneously subjected to the previously mentioned storage temperatures and sampled biweekly for their total bacterial count (TPC) and yeasts/molds (Y/M) count. During the 2-week short-term storage, L. monocytogenes population on organic Granny Smith apples stored at 1, 4, or 10°C was reduced by 0.2-0.3 Log. When apples were stored at 22°C, there was a 0.5-1.2 Log 10 CFU/apple reduction 14-day post storage dependent on the initial inoculation level. During the 12-week cold storage under 1, 4, and 10°C, L. monocytogenes count on organic Granny Smith apples decreased by 0.5-1.5 Log 10 CFU/apple for both inoculation levels. L. monocytogenes had similar survival pattern on conventional Granny Smith and Fuji apples with 0.8-2.0 Log 10 CFU/apple reduction over a 3-month cold storage period. Interestingly, both TPC and Y/M count were stable regardless of apple variety or cultivation practice during the 12-week storage at all tested temperatures. In summary, while L. monocytogenes did not proliferate on apple surfaces during 12 weeks of refrigerated storage, only a limited reduction of L. monocytogenes was observed in this study. Therefore, the apple industry cannot rely on cold storage alone to control this pathogen. Additional interventions are needed to eradicate Listeria on fresh apples during long-term cold storage.

Microbial effect of steam vacuum pasteurisation implemented after slaughtering and dressing of sheep and lamb.

PubMed

Hassan, Ammar Ali; Skjerve, Eystein; Bergh, Claus; Nesbakken, Truls

2015-01-01

The main objective of the study was to assess the effect of steam vacuum pasteurisation on carcass contamination with focus on Escherichia coli, Enterobacteriaceae and total plate count (TPC). Additionally, the effect of an additional tryptone soy agar (TSA) step for resuscitation of Enterobacteriaceae after steam vacuum pasteurisation was investigated. Steam vacuum pasteurisation was applied at a temperature of >82°C for a duration of 10s on sheep and lamb carcasses (n=120). Samples were taken immediately: i) after trimming just before the use of steam vacuum and ii) after use of steam vacuum. Nordic Committee on Food Analysis methods were used in microbial analyses. The differences in log reduction were found significant for all of the three microorganisms (p<0.05). For TPC, the general reduction was a 0.65 log10 in the number of colony forming units (CFU) per cm(2). For E. coli, the median reduction effect on carcasses positive before decontamination was 1.1 log10 CFU/cm(2). A large variability of the effect was however found, with 50% of the figures ranging from a 0.24 to 1.62 log10 CFU/cm(2) reduction and a 10-90% range of 0-2.1. The number of positive carcasses with Enterobacteriaceae after steam vacuum pasteurisation was higher in samples where TSA+violet red bile glucose agar (VRBGA) was used compared to samples where only VRBGA was used (p<0.01). Steam vacuum pasteurisation was found efficient in reducing the total count, read as TPC, as well as the level of E. coli and Enterobacteriaceae. Copyright © 2014 Elsevier Ltd. All rights reserved.

Listeria monocytogenes Internalizes in Romaine Lettuce Grown in Greenhouse Conditions.

PubMed

Shenoy, Archana G; Oliver, Haley F; Deering, Amanda J

2017-04-01

Listeria monocytogenes has been implicated in a number of outbreaks involving fresh produce, including an outbreak in 2016 resulting from contaminated packaged salads. The persistence and internalization potential of L. monocytogenes in romaine lettuce was evaluated, and the persistence of two L. monocytogenes strains was assessed on three romaine lettuce cultivars. Seeds were germinated, and plants grown in three soil types (i.e., standard potting mix, autoclaved potting mix, and top soil) and sterile soft-top agar for up to 21 days. Average CFU per gram of L. monocytogenes on seeds and plants was calculated from five replicates per harvest day. Up to 8.2 log CFU/g L. monocytogenes persisted on romaine lettuce plants (Braveheart cultivar) grown in soft-top agar, while those grown in commercial potting mix (initial soil aerobic plate count of 4.0 × 10 4 CFU/g) had a final concentration of 5.4 log CFU/g, and autoclaved commercial potting mix had a final concentration of 3.8 ± 0.2 log CFU/g after a 21-day period. Pathogen levels dropped below the limit of detection (2 log CFU/g) by day 18 in 75% topsoil (initial soil aerobic plate count of 4.0 × 10 1 CFU/g); this did not occur in sterile media. Although L. monocytogenes strain differences and presence of a clay coating on seeds did not affect persistence, differences were observed in L. monocytogenes growth and survival among cultivars. To assess internalization, seeds were inoculated with L. monocytogenes expressing green fluorescent protein. Three plants were fixed, paraffin embedded, and sectioned; localization was studied by using standard immunohistochemistry techniques. A total of 539 internalized L. monocytogenes cells were visualized among three 20-day seedlings. L. monocytogenes cells were located in all major tissue types (pith followed by cortex, xylem, phloem, and epidermis). The presence of L. monocytogenes in the plant vasculature suggests potential for transport throughout the plant into edible tissue.

High pressure homogenization vs heat treatment: safety and functional properties of liquid whole egg.

PubMed

Patrignani, Francesca; Vannini, Lucia; Sado Kamdem, Sylvain L; Hernando, Isabel; Marco-Molés, Raquel; Guerzoni, M Elisabetta; Lanciotti, Rosalba

2013-10-01

This research investigated the potential of multi-pass homogenization treatment for the inactivation of Salmonella enterica serovar Enteritidis inoculated at different levels in liquid whole egg (LWE) comparing the efficacy of this treatment with a traditional thermal one performed at 65 °C. Moreover, the effects of high pressure treatment (HPH) on structural and functional properties such as viscosity, microstructure and foaming abilities of LWE were investigated. The data obtained suggested that the multi-pass high pressure treatment at 100 MPa of S. enterica serovar Enteritidis inoculated in LWE at 7 and 4 log CFU/ml resulted in a first order inactivation kinetic, while the thermal inactivation curves of S. enterica serovar Enteritidis inoculated at 8 and 4 log CFU/ml presented a non-linear behaviour, with a marked tail after 3 min of treatment at 65 °C. Additionally, HPH treatment caused an increase in foaming capacity of LWE, with respect to the untreated samples, passing from values of 26% of the control to 50% of pressure treated samples. Copyright © 2013 Elsevier Ltd. All rights reserved.

Antimicrobial efficacy and wound-healing property of a topical ointment containing nitric-oxide-loaded zeolites

PubMed Central

Ercan, Utku K.; Bhattacharyya, Aparna; Samuels, Joshua; Sedlak, Jason; Trikha, Ritika; Barbee, Kenneth A.; Weingarten, Michael S.

2014-01-01

Topical delivery of nitric oxide (NO) through a wound dressing has the potential to reduce wound infections and improve healing of acute and chronic wounds. This study characterized the antibacterial efficacy of an ointment containing NO-loaded, zinc-exchanged zeolite A that releases NO upon contact with water. The release rate of NO from the ointment was measured using a chemiluminescence detection system. Minimum bactericidal concentration assays were performed using five common wound pathogens, including Gram-negative bacteria (Escherichia coli and Acinetobacter baumannii), Gram-positive bacteria (Staphylococcus epidermidis and meticillin-resistant Staphylococcus aureus) and a fungus (Candida albicans). The time dependence of antimicrobial activity was characterized by performing log-reduction assays at four time points after 1–8 h ointment exposure. The cytotoxicity of the ointment after 24 h was assessed using cultured 3T3 fibroblast cells. Minimum microbicidal concentrations (MMCs) for bacterial organisms (5×107 c.f.u.) ranged from 50 to 100 mg ointment (ml media)−1; the MMC for C. albicans (5×104 c.f.u.) was 50 mg ointment (ml media)−1. Five to eight log reductions in bacterial viability and three log reductions in fungal viability were observed after 8 h exposure to NO–zeolite ointment compared with untreated organisms. Fibroblasts remained viable after 24 h exposure to the same concentration of NO–zeolite ointment as was used in antimicrobial tests. In parallel studies, full-thickness cutaneous wounds on Zucker obese rats healed faster than wounds treated with a control ointment. These data indicate that ointment containing NO-loaded zeolites could potentially be used as a broad-spectrum antimicrobial wound-healing dressing. PMID:24196133

Thermal inactivation of Escherichia coli O157:H7 in blade-tenderized beef steaks cooked on a commercial open-flame gas grill.

PubMed

Luchansky, John B; Porto-Fett, Anna C S; Shoyer, Bradley; Phebus, Randall K; Thippareddi, Harshavardhan; Call, Jeffrey E

2009-07-01

Beef subprimals were inoculated on the lean side with ca. 4.0 log CFU/g of a cocktail of rifampin-resistant (Rif(r)) Escherichia coli O157:H7 strains and then passed once through a mechanical blade tenderizer with the lean side facing upward. Inoculated subprimals that were not tenderized served as controls. Two core samples were removed from each of three tenderized subprimals and cut into six consecutive segments starting from the inoculated side. A total of six cores were also obtained from control subprimals, but only segment 1 (topmost) was sampled. Levels of E. coli O157:H7 recovered from segment 1 were 3.81 log CFU/g for the control subprimals and 3.36 log CFU/g for tenderized subprimals. The percentage of cells recovered in segment 2 was ca. 25-fold lower than levels recovered from segment 1, but E. coli O157:H7 was recovered from all six segments of the cores obtained from tenderized subprimals. In phase II, lean-side-inoculated (ca. 4.0 log CFU/g), single-pass tenderized subprimals were cut into steaks of various thicknesses (1.91 cm [0.75 in.], 2.54 cm [1.0 in.], and 3.18 cm [1.25 in.]) that were subsequently cooked on a commercial open-flame gas grill to internal temperatures of 48.8 degrees C (120 degrees F), 54.4 degrees C (130 degrees F), and 60 degrees C (140 degrees F). In general, regardless of temperature or thickness, we observed about a 2.6- to 4.2-log CFU/g reduction in pathogen levels following cooking. These data validate that cooking on a commercial gas grill is effective at eliminating relatively low levels of the pathogen that may be distributed throughout a blade-tenderized steak.

Incidence and behavior of Salmonella and Escherichia coli on whole and sliced zucchini squash (Cucurbitapepo) fruit.

PubMed

Castro-Rosas, Javier; Santos López, Eva María; Gómez-Aldapa, Carlos Alberto; González Ramírez, Cesar Abelardo; Villagomez-Ibarra, José Roberto; Gordillo-Martínez, Alberto José; López, Angélica Villarruel; del Refugio Torres-Vitela, M

2010-08-01

The incidence of coliform bacteria (CB), thermotolerant coliforms (TC), Escherichia coli, and Salmonella was determined for zucchini squash fruit. In addition, the behavior of four serotypes of Salmonella and a cocktail of three E. coli strains on whole and sliced zucchini squash at 25+/-2 degrees C and 3 to 5 degrees C was tested. Squash fruit was collected in the markets of Pachuca city, Hidalgo State, Mexico. CB, TC, E. coli, and Salmonella were detected in 100, 70, 62, and 10% of the produce, respectively. The concentration ranged from 3.8 to 7.4 log CFU per sample for CB, and >3 to 1,100 most probable number per sample for TC and E. coli. On whole fruit stored at 25+/-2 degrees C or 3 to 5 degrees C, no growth was observed for any of the tested microorganisms or cocktails thereof. After 15 days at 25+/-2 degrees C, the tested Salmonella serotypes had decreased from an initial inoculum level of 7 log CFU to <1 log, and at 3 to 5 degrees C they decreased to approximately 2 log. Survival of E. coli was significantly greater than for the Salmonella strains at the same times and temperatures; after 15 days, at 25+/-2 degrees C E. coli cocktail strains had decreased to 3.4 log CFU per fruit and at 3 to 5 degrees C they decreased to 3.6 log CFU per fruit. Both the Salmonella serotypes and E. coli strains grew when inoculated onto sliced squash: after 24 h at 25+/-2 degrees C, both bacteria had grown to approximately 6.5 log CFU per slice. At 3 to 5 degrees C, the bacterial growth was inhibited. The squash may be an important factor contributing to the endemicity of Salmonella in Mexico.

Establishment of the first international repository for transfusion-relevant bacteria reference strains: ISBT working party transfusion-transmitted infectious diseases (WP-TTID), subgroup on bacteria.

PubMed

Störmer, M; Arroyo, A; Brachert, J; Carrero, H; Devine, D; Epstein, J S; Gabriel, C; Gelber, C; Goodrich, R; Hanschmann, K-M; Heath, D G; Jacobs, M R; Keil, S; de Korte, D; Lambrecht, B; Lee, C-K; Marcelis, J; Marschner, S; McDonald, C; McGuane, S; McKee, M; Müller, T H; Muthivhi, T; Pettersson, A; Radziwon, P; Ramirez-Arcos, S; Reesink, H W; Rojo, J; Rood, I; Schmidt, M; Schneider, C K; Seifried, E; Sicker, U; Wendel, S; Wood, E M; Yomtovian, R A; Montag, T

2012-01-01

Bacterial contamination of platelet concentrates (PCs) still remains a significant problem in transfusion with potential important clinical consequences, including death. The International Society of Blood Transfusion Working Party on Transfusion-Transmitted Infectious Diseases, Subgroup on Bacteria, organised an international study on Transfusion-Relevant Bacteria References to be used as a tool for development, validation and comparison of both bacterial screening and pathogen reduction methods. Four Bacteria References (Staphylococcus epidermidis PEI-B-06, Streptococcus pyogenes PEI-B-20, Klebsiella pneumoniae PEI-B-08 and Escherichia coli PEI-B-19) were selected regarding their ability to proliferate to high counts in PCs and distributed anonymised to 14 laboratories in 10 countries for identification, enumeration and bacterial proliferation in PCs after low spiking (0·3 and 0·03 CFU/ml), to simulate contamination occurring during blood donation. Bacteria References were correctly identified in 98% of all 52 identifications. S. pyogenes and E. coli grew in PCs in 11 out of 12 laboratories, and K. pneumoniae and S. epidermidis replicated in all participating laboratories. The results of bacterial counts were very consistent between laboratories: the 95% confidence intervals were for S. epidermidis: 1·19-1·32 × 10(7) CFU/ml, S. pyogenes: 0·58-0·69 × 10(7) CFU/ml, K. pneumoniae: 18·71-20·26 × 10(7) CFU/ml and E. coli: 1·78-2·10 × 10(7) CFU/ml. The study was undertaken as a proof of principle with the aim to demonstrate (i) the quality, stability and suitability of the bacterial strains for low-titre spiking of blood components, (ii) the property of donor-independent proliferation in PCs, and (iii) their suitability for worldwide shipping of deep frozen, blinded pathogenic bacteria. These aims were successfully fulfilled. The WHO Expert Committee Biological Standardisation has approved the adoption of these four bacteria strains as the first Repository for Transfusion-Relevant Bacteria Reference Strains and, additionally, endorsed as a project the addition of six further bacteria strain preparations suitable for control of platelet contamination as the next step of enlargement of the repository. © 2011 The Author(s). Vox Sanguinis © 2011 International Society of Blood Transfusion.

Campylobacter spp. contamination of chicken carcasses during processing in relation to flock colonisation.

PubMed

Allen, V M; Bull, S A; Corry, J E L; Domingue, G; Jørgensen, F; Frost, J A; Whyte, R; Gonzalez, A; Elviss, N; Humphrey, T J

2007-01-01

The presence and numbers of campylobacters on chicken carcasses from 26 slaughter groups, originating from 22 single-house flocks and processed in four UK plants, were studied in relation to the level of flock colonisation determined by examining the caecal contents of at least ten birds per group. The prevalence of campylobacters on carcasses from five campylobacter-negative flocks processed just after other negative flocks was low (8.0 log(10) cfu) than carcasses originating from low prevalence flocks (average of 2.3 log(10) cfu; range: <1.1 to 4.1 log(10) cfu). There was a reduction in the numbers of campylobacters on carcasses between plucking and chilling in eight of ten fully colonised flocks. In another eight flocks, a significant (P<0.001) decrease (0.8 log(10) cfu) in the number of campylobacters on carcasses from just before to after chilling was detected. Campylobacter spp. could be isolated from aerosols, particles and droplets in considerable numbers in the hanging-on, defeathering and evisceration areas but not in the chillers. This was the case even when campylobacters were not isolated from the target flock. Campylobacters on carcasses from two partly colonised flocks were either the same subtype, as determined by speciation, Multi-Locus Sequence Typing (MLST) and flaA Restricted Fragment Length Polymorphism (RFLP) typing, as those in the fully colonised flocks processed previously, although not necessarily the most prevalent ones; or were the same subtypes as those found in the caeca of the flock itself. The prevalences of the different campylobacter subtypes found on carcasses from two fully colonised flocks did not closely reflect those found in the caeca. MLST combined with flaA RFLP provided a good method for ascertaining the relatedness of strains isolated from carcasses and caecal contents. This study showed that carcass contamination is related to the within-flock prevalence of campylobacter colonisation, but that contamination from previously processed flocks was also significant, especially on carcasses from low prevalence flocks. Forced dry air cooling of carcasses reduced contamination levels.

Inactivation of Bacillus anthracis Spores by a Combination of Biocides and Heating under High-Temperature Short-Time Pasteurization Conditions ▿

PubMed Central

Xu, Sa; Labuza, Theodore P.; Diez-Gonzalez, Francisco

2008-01-01

The milk supply is considered a primary route for a bioterrorism attack with Bacillus anthracis spores because typical high-temperature short-time (HTST) pasteurization conditions cannot inactivate spores. In the event of intentional contamination, an effective method to inactivate the spores in milk under HTST processing conditions is needed. This study was undertaken to identify combinations and concentrations of biocides that can inactivate B. anthracis spores at temperatures in the HTST range in less than 1 min. Hydrogen peroxide (HP), sodium hypochlorite (SH), and peroxyacetic acid (PA) were evaluated for their efficacy in inactivating spores of strains 7702, ANR-1, and 9131 in milk at 72, 80, and 85°C using a sealed capillary tube technique. Strains ANR-1 and 9131 were more resistant to all of the biocide treatments than strain 7702. Addition of 1,260 ppm SH to milk reduced the number of viable spores of each strain by 6 log CFU/ml in less than 90 and 60 s at 72 and 80°C, respectively. After neutralization, 1,260 ppm SH reduced the time necessary to inactivate 6 log CFU/ml (TTI6-log) at 80°C to less than 20 s. Treatment of milk with 7,000 ppm HP resulted in a similar level of inactivation in 60 s. Combined treatment with 1,260 ppm SH and 1,800 ppm HP inactivated spores of all strains in less than 20 s at 80°C. Mixing 15 ppm PA with milk containing 1,260 ppm SH resulted in TTI6-log of 25 and 12 s at 72 and 80°C, respectively. TTI6-log of less than 20 s were also achieved at 80°C by using two combinations of biocides: 250 ppm SH, 700 ppm HP, and 150 ppm PA; and 420 ppm SH (pH 7), 1,100 ppm HP, and 15 ppm PA. These results indicated that different combinations of biocides could consistently result in 6-log reductions in the number of B. anthracis spores in less than 1 min at temperatures in the HTST range. This information could be useful for developing more effective thermal treatment strategies which could be used in HTST milk plants to process contaminated milk for disposal and decontamination, as well as for potential protective measures. PMID:18390680

Inactivation of Bacillus anthracis spores by a combination of biocides and heating under high-temperature short-time pasteurization conditions.

PubMed

Xu, Sa; Labuza, Theodore P; Diez-Gonzalez, Francisco

2008-06-01

The milk supply is considered a primary route for a bioterrorism attack with Bacillus anthracis spores because typical high-temperature short-time (HTST) pasteurization conditions cannot inactivate spores. In the event of intentional contamination, an effective method to inactivate the spores in milk under HTST processing conditions is needed. This study was undertaken to identify combinations and concentrations of biocides that can inactivate B. anthracis spores at temperatures in the HTST range in less than 1 min. Hydrogen peroxide (HP), sodium hypochlorite (SH), and peroxyacetic acid (PA) were evaluated for their efficacy in inactivating spores of strains 7702, ANR-1, and 9131 in milk at 72, 80, and 85 degrees C using a sealed capillary tube technique. Strains ANR-1 and 9131 were more resistant to all of the biocide treatments than strain 7702. Addition of 1,260 ppm SH to milk reduced the number of viable spores of each strain by 6 log CFU/ml in less than 90 and 60 s at 72 and 80 degrees C, respectively. After neutralization, 1,260 ppm SH reduced the time necessary to inactivate 6 log CFU/ml (TTI6-log) at 80 degrees C to less than 20 s. Treatment of milk with 7,000 ppm HP resulted in a similar level of inactivation in 60 s. Combined treatment with 1,260 ppm SH and 1,800 ppm HP inactivated spores of all strains in less than 20 s at 80 degrees C. Mixing 15 ppm PA with milk containing 1,260 ppm SH resulted in TTI6-log of 25 and 12 s at 72 and 80 degrees C, respectively. TTI6-log of less than 20 s were also achieved at 80 degrees C by using two combinations of biocides: 250 ppm SH, 700 ppm HP, and 150 ppm PA; and 420 ppm SH (pH 7), 1,100 ppm HP, and 15 ppm PA. These results indicated that different combinations of biocides could consistently result in 6-log reductions in the number of B. anthracis spores in less than 1 min at temperatures in the HTST range. This information could be useful for developing more effective thermal treatment strategies which could be used in HTST milk plants to process contaminated milk for disposal and decontamination, as well as for potential protective measures.

Survival of Lactobacillus acidophilus and Bifidobacterium bifidum in ice cream for use as a probiotic food.

PubMed

Hekmat, S; McMahon, D J

1992-06-01

Probiotic ice cream was made by fermenting a standard ice cream mix with Lactobacillus acidophilus and Bifidobacterium bifidum cultures and then freezing the mix in a batch freezer. Survival of the L. acidophilus and B. bifidum, as well as beta-galactosidase activity, was monitored during 17 wk of frozen storage at -29 degrees C. After freezing of the fermented mix, bacterial counts were 1.5 x 10(8) cfu/ml for L. acidophilus and 2.5 x 10(8) cfu/ml for B. bifidum. Seventeen weeks after freezing, these counts had decreased to 4 x 10(6) and 1 x 10(7) cfu/ml, respectively. During the same period, beta-galactosidase activity decreased from 1800 to 1300 units/ml. Probiotic ice cream was prepared at pH 5.0, 5.5, and 6.0 to determine consumer preferences and was compared with standard Utah State University "Aggie" ice cream. All samples were strawberry-flavored and were evaluated by 88 judges. The preferred pH of probiotic ice cream, based on overall acceptance, was pH 5.5. We demonstrated that probiotic ice cream is a suitable vehicle for delivering beneficial microorganisms such as L. acidophilus and B. bifidum to consumers. The bacteria can be grown to high numbers in ice cream mix and remain viable during frozen storage.

Antimicrobial effect of herb extracts against Escherichia coli O157:H7, Listeria monocytogenes, and Salmonella typhimurium associated with beef.

PubMed

Cutter, C N

2000-05-01

The effects of plant extracts against pathogenic bacteria in vitro are well known, yet few studies have addressed the effects of these compounds against pathogens associated with muscle foods. A series of experiments was conducted to determine the effectiveness of a commercially available, generally recognized as safe, herb extract dispersed in sodium citrate (Protecta One) or sodium chloride (Protecta Two) against Escherichia coli O157:H7, Salmonella typhimurium, and Listeria monocytogenes associated with beef. In the first experiment, E. coli O157:H7, Salmonella typhimurium, and L. monocytogenes inoculated onto beef and subjected to surface spray treatments with 2.5% solutions of Protecta One or Protecta Two were not affected by immediate application (day 0) of the herbal extracts. However, after 7 days of storage at 4 degrees C, E. coli O157:H7 was reduced by >1.3 log10 CFU/cm2 by Protecta Two; L. monocytogenes was reduced by 1.8 and 1.9 log10 CFU/cm2 by Protecta One and Protecta Two, respectively; Salmonella typhimurium was not reduced >0.3 log10 CFU/cm2 by either extract by day 7. In the second experiment, 2.5% Protecta Two (wt/vol or wt/wt) added to inoculated lean and adipose beef trim, processed, and packaged as ground beef chubs (80% lean, 20% adipose), did not reduce pathogen populations >0.5 log10 CFU/g up to 14 days at 4 degrees C. In the third experiment, surface spray treatments of beef with 2.5% lactic acid or 2.5% solutions of Protecta One or Protecta Two, vacuum packaged, and stored up to 35 days at 4 degrees C did reduce E. coli O157:H7, L. monocytogenes, and Salmonella Typhimurium slightly. These studies suggest that the use of herb extracts may afford some reductions of pathogens on beef surfaces; however, the antimicrobial activity may be diminished in ground beef by adipose components.

Survival of cold-stressed Campylobacter jejuni on ground chicken and chicken skin during frozen storage.

PubMed

Bhaduri, Saumya; Cottrell, Bryan

2004-12-01

Campylobacter jejuni is prevalent in poultry, but the effect of combined refrigerated and frozen storage on its survival, conditions relevant to poultry processing and storage, has not been evaluated. Therefore, the effects of refrigeration at 4 degrees C, freezing at -20 degrees C, and a combination of refrigeration and freezing on the survival of C. jejuni in ground chicken and on chicken skin were examined. Samples were enumerated using tryptic soy agar containing sheep's blood and modified cefoperazone charcoal deoxycholate agar. Refrigerated storage alone for 3 to 7 days produced a reduction in cell counts of 0.34 to 0.81 log10 CFU/g in ground chicken and a reduction in cell counts of 0.31 to 0.63 log10 CFU/g on chicken skin. Declines were comparable for each sample type using either plating medium. Frozen storage, alone and with prerefrigeration, produced a reduction in cell counts of 0.56 to 1.57 log10 CFU/g in ground chicken and a reduction in cell counts of 1.38 to 3.39 log10 CFU/g on chicken skin over a 2-week period. The recovery of C. jejuni following freezing was similar on both plating media. The survival following frozen storage was greater in ground chicken than on chicken skin with or without prerefrigeration. Cell counts after freezing were lower on chicken skin samples that had been prerefrigerated for 7 days than in those that had been prerefrigerated for 0, 1, or 3 days. This was not observed for ground chicken samples, possibly due to their composition. C. jejuni survived storage at 4 and -20 degrees C with either sample type. This study indicates that, individually or in combination, refrigeration and freezing are not a substitute for safe handling and proper cooking of poultry.

Performance Characteristics of the QUANTIPLEX HIV-1 RNA 3.0 Assay for Detection and Quantitation of Human Immunodeficiency Virus Type 1 RNA in Plasma

PubMed Central

Erice, Alejo; Brambilla, Donald; Bremer, James; Jackson, J. Brooks; Kokka, Robert; Yen-Lieberman, Belinda; Coombs, Robert W.

2000-01-01

The QUANTIPLEX HIV-1 RNA assay, version 3.0 (a branched DNA, version 3.0, assay [bDNA 3.0 assay]), was evaluated by analyzing spiked and clinical plasma samples and was compared with the AMPLICOR HIV-1 MONITOR Ultrasensitive (ultrasensitive reverse transcription-PCR [US-RT-PCR]) method. A panel of spiked plasma samples that contained 0 to 750,000 copies of human immunodeficiency virus type 1 (HIV-1) RNA per ml was tested four times in each of four laboratories (1,344 assays). Negative results (<50 copies/ml) were obtained in 30 of 32 (94%) assays with seronegative samples, 66 of 128 (52%) assays with HIV-1 RNA at 50 copies/ml, and 5 of 128 (4%) assays with HIV-1 RNA at 100 copies/ml. The assay was linear from 100 to 500,000 copies/ml. The within-run standard deviation (SD) of the log10 estimated HIV-1 RNA concentration was 0.08 at 1,000 to 500,000 copies/ml, increased below 1,000 copies/ml, and was 0.17 at 100 copies/ml. Between-run reproducibility at 100 to 500 copies/ml was <0.10 log10 in most comparisons. Interlaboratory differences across runs were ≤0.10 log10 at all concentrations examined. A subset of the panel (25 to 500 copies/ml) was also analyzed by the US-RT-PCR assay. The within-run SD varied inversely with the log10 HIV-1 RNA concentration but was higher than the SD for the bDNA 3.0 assay at all concentrations. Log-log regression analysis indicated that the two methods produced very similar estimates at 100 to 500 copies/ml. In parallel testing of clinical specimens with low HIV-1 RNA levels, 80 plasma samples with <50 copies/ml by the US-RT-PCR assay had <50 copies/ml when they were retested by the bDNA 3.0 assay. In contrast, 11 of 78 (14%) plasma samples with <50 copies/ml by the bDNA 3.0 assay had ≥50 copies/ml when they were retested by the US-RT-PCR assay (median, 86 copies/ml; range, 50 to 217 copies/ml). Estimation of bDNA 3.0 values of <50 copies/ml by extending the standard curve of the assay showed that these samples with discrepant results had higher HIV-1 RNA levels than the samples with concordant results (median, 34 versus 17 copies/ml; P = 0.0051 by the Wilcoxon two-sample test). The excellent reproducibility, broad linear range, and good sensitivity of the bDNA 3.0 assay make it a very attractive method for quantitation of HIV-1 RNA levels in plasma. PMID:10921936

Biofilm growth on polyvinylchloride surface incubated in suboptimal microbial warm water and effect of sanitizers on biofilm removal post biofilm formation.

PubMed

Maharjan, Pramir; Huff, Geraldine; Zhang, Wen; Watkins, Susan

2017-01-01

An in vitro experiment was conducted to understand the nature of biofilm growth on polyvinyl chloride (PVC) surface when exposed to suboptimal-quality microbial water (>4 log 10 cfu/mL) obtained from a poultry drinking water source mimicking water in waterlines during the first week of poultry brooding condition. PVC sections (internal surface area of 15.16 cm 2 ) were utilized in the study to grow biofilm. After a 7-d test period, test coupons with 7-day-old biofilm were transferred into autoclaved municipal water and then treated with either chlorine-based or hydrogen peroxide-based sanitizer at bird drinking water rate, to see the impact on removal of biofilm formed on test coupons. Two trials (T1 and T2) were conducted. Test coupons used in T1 and T2 had the bacterial growth of 3.67 (SEM 0.04) and 3.97 (SEM 0.11) log 10 cfu/cm 2 on d 7. After sanitizer application, chlorine-based sanitizer removed bacteria in biofilm completely (0 cfu/cm 2 ) within 24 h post treatment whereas hydrogen peroxide-based sanitizer reduced the counts to 1.68 log 10 cfu/cm 2 (P < 0.05) by 48 h post sanitizer application. Control remained the same (P > 0.05). Results indicated that biofilm formation can occur quickly under suboptimal water condition on PVC surface, and sanitizer application helped mitigate already formed biofilm, yet chlorine proved to be more effective than hydrogen peroxide. © 2016 Poultry Science Association Inc.

Intra- and Interlaboratory Variabilities of Results Obtained with the Quantiplex Human Immunodeficiency Virus Type 1 RNA bDNA Assay, Version 3.0

PubMed Central

Kellogg, James A.; Atria, Peter V.; Sanders, Jeffrey C.; Eyster, M. Elaine

2001-01-01

Normal assay variation associated with bDNA tests for human immunodeficiency virus type 1 (HIV-1) RNA performed at two laboratories with different levels of test experience was investigated. Two 5-ml aliquots of blood in EDTA tubes were collected from each patient for whom the HIV-1 bDNA test was ordered. Blood was stored for no more than 4 h at room temperature prior to plasma separation. Plasma was stored at −70°C until transported to the Central Pennsylvania Alliance Laboratory (CPAL; York, Pa.) and to the Hershey Medical Center (Hershey, Pa.) on dry ice. Samples were stored at ≤−70°C at both laboratories prior to testing. Pools of negative (donor), low-HIV-1-RNA-positive, and high-HIV-1-RNA-positive plasma samples were also repeatedly tested at CPAL to determine both intra- and interrun variation. From 11 August 1999 until 14 September 2000, 448 patient specimens were analyzed in parallel at CPAL and Hershey. From 206 samples with results of ≥1,000 copies/ml at CPAL, 148 (72%) of the results varied by ≤0.20 log10 when tested at Hershey and none varied by >0.50 log10. However, of 242 specimens with results of <1,000 copies/ml at CPAL, 11 (5%) of the results varied by >0.50 log10 when tested at Hershey. Of 38 aliquots of HIV-1 RNA pool negative samples included in 13 CPAL bDNA runs, 37 (97%) gave results of <50 copies/ml and 1 (3%) gave a result of 114 copies/ml. Low-positive HIV-1 RNA pool intrarun variation ranged from 0.06 to 0.26 log10 while the maximum interrun variation was 0.52 log10. High-positive HIV-1 RNA pool intrarun variation ranged from 0.04 to 0.32 log10, while the maximum interrun variation was 0.55 log10. In our patient population, a change in bDNA HIV-1 RNA results of ≤0.50 log10 over time most likely represents normal laboratory test variation. However, a change of >0.50 log10, especially if the results are >1,000 copies/ml, is likely to be significant. PMID:11329458

Intra- and interlaboratory variabilities of results obtained with the Quantiplex human immunodeficiency virus type 1 RNA bDNA assay, version 3.0.

PubMed

Kellogg, J A; Atria, P V; Sanders, J C; Eyster, M E

2001-05-01

Normal assay variation associated with bDNA tests for human immunodeficiency virus type 1 (HIV-1) RNA performed at two laboratories with different levels of test experience was investigated. Two 5-ml aliquots of blood in EDTA tubes were collected from each patient for whom the HIV-1 bDNA test was ordered. Blood was stored for no more than 4 h at room temperature prior to plasma separation. Plasma was stored at -70 degrees C until transported to the Central Pennsylvania Alliance Laboratory (CPAL; York, Pa.) and to the Hershey Medical Center (Hershey, Pa.) on dry ice. Samples were stored at < or =-70 degrees C at both laboratories prior to testing. Pools of negative (donor), low-HIV-1-RNA-positive, and high-HIV-1-RNA-positive plasma samples were also repeatedly tested at CPAL to determine both intra- and interrun variation. From 11 August 1999 until 14 September 2000, 448 patient specimens were analyzed in parallel at CPAL and Hershey. From 206 samples with results of > or =1,000 copies/ml at CPAL, 148 (72%) of the results varied by < or =0.20 log(10) when tested at Hershey and none varied by >0.50 log(10). However, of 242 specimens with results of <1,000 copies/ml at CPAL, 11 (5%) of the results varied by >0.50 log(10) when tested at Hershey. Of 38 aliquots of HIV-1 RNA pool negative samples included in 13 CPAL bDNA runs, 37 (97%) gave results of <50 copies/ml and 1 (3%) gave a result of 114 copies/ml. Low-positive HIV-1 RNA pool intrarun variation ranged from 0.06 to 0.26 log(10) while the maximum interrun variation was 0.52 log(10). High-positive HIV-1 RNA pool intrarun variation ranged from 0.04 to 0.32 log(10), while the maximum interrun variation was 0.55 log(10). In our patient population, a change in bDNA HIV-1 RNA results of < or =0.50 log(10) over time most likely represents normal laboratory test variation. However, a change of >0.50 log(10), especially if the results are >1,000 copies/ml, is likely to be significant.

Antimicrobial treatments to control Listeria monocytogenes in queso fresco.

PubMed

Lourenço, António; Kamnetz, Mary B; Gadotti, Camila; Diez-Gonzalez, Francisco

2017-06-01

Queso fresco, is a Hispanic non-fermented cheese highly susceptible to contamination with L. monocytogenes. This research was aimed to determine the effect of GRAS antimicrobial ingredients to control L. monocytogenes. Antimicrobials included caprylic acid (CA), Nisaplin ® (N, 2.5% nisin), a mixture of sodium lactate and sodium diacetate (SL/SD), Lactococcus lactis sbp. lactis DPC 3147, monolaurin, and lactic acid (LA). Batches of queso fresco curds were inoculated with 10 4  CFU/g and stored at 4 °C for three weeks. During storage the count of L. monocytogenes reached 7 to 8 Log CFU/g in control samples. Most individual antimicrobial treatments resulted in less than 1 Log CFU/g reductions in final counts, with the exception of N (0.5 g/kg) and CA (2.9 g/kg) that caused more than 3 and 5 Log CFU/g differences with controls, respectively. Mixtures of ingredients were more effective in inhibiting L. monocytogenes growth, and treatments with N and CA consistently delivered 6 Log CFU/g less counts than controls. Supplementation of 12 g/kg LA to treatments with SL/SD (3%/0.22%) caused differences of more than 4 Log CFU/g in final Listeria populations. Samples treated with the binary mixtures of N and CA (0.5 and 0.7 g/kg, respectively) were evaluated in a consumer panel (n = 67). Panelists slightly preferred control and commercial over treated samples, but all samples were in average rated between "slightly liking" and "moderately liking." These experiments indicated that combined use of antimicrobial ingredients may be an effective way to control the population of Listeria monocytogenes in queso fresco. Copyright © 2016 Elsevier Ltd. All rights reserved.

Sequential disinfection of E. coli O157:H7 on shredded lettuce leaves by aqueous chlorine dioxide, ozonated water, and thyme essential oil

NASA Astrophysics Data System (ADS)

Singh, Nepal; Singh, Rakesh K.; Bhunia, Arun K.; Stroshine, Richard L.; Simon, James E.

2001-03-01

There have been numerous studies on effectiveness of different sanitizers for microbial inactivation. However, results obtained from different studies indicate that microorganism cannot be easily removed from fresh cut vegetables because of puncture and cut surfaces with varying surface topographies. In this study, three step disinfection approach was evaluated for inactivation of E. coli O157:H7 on shredded lettuce leaves. Sequential application of thyme oil, ozonated water, and aqueous chlorine dioxide was evaluated in which thyme oil was applied first followed by ozonated water and aqueous chlorine dioxide. Shredded lettuce leaves inoculated with cocktail culture of E. coli O157:H7 (C7927, EDL 933 and 204 P), were washed with ozonated water (15 mg/l for 10min), aqueous chlorine dioxide (10 mg/l,for 10min) and thyme oil suspension (0.1%, v/v for 5min). Washing of lettuce leaves with ozonated water, chlorine dioxide and thyme oil suspension resulted in 0.44, 1.20, and 1.46 log reduction (log10 cfu/g), respectively. However, the sequential treatment achieved approximately 3.13 log reductions (log10 cfu/g). These results demonstrate the efficacy of sequential treatments in decontaminating shredded lettuce leaves containing E. coli O157:H7.

Inactivation effect of X-ray treatments on Cronobacter species (Enterobacter sakazakii) in tryptic soy broth, skim milk, low-fat milk and whole-fat milk.

PubMed

Mahmoud, B S M

2009-11-01

To determine the inactivation effect of X-ray treatments on Cronobacter (E. sakazakii) in tryptic soy broth (TSB), skim milk (0% fat), low-fat milk (1% and 2%) and whole-fat milk (3.5%). X-rays were produced using the RS 2400 generator system (Rad Source Technologies Inc.). Cronobacter (in TSB), inoculated skim milk (0% fat), low-fat milk (1% and 2% fat) and whole-fat milk (3.5% fat) were treated with 0.0, 0.1, 0.5, 0.75, 1.0, 2.0, 3.0, 4.0, 5.0 and 6.0 kGy X-ray doses. Surviving bacteria in the TSB and inoculated milk, before and after treatment, were enumerated using plating method onto trypticase soy agar. Greater than 7.0-log CFU reduction in Cronobacter population was observed with 4.0, 5.0, 6.0, 6.0 and 6.0 kGy X-ray in the TSB, skim milk, 1% fat milk, 2% fat milk and 3.5% fat milk, respectively. Treatment with X-rays significantly (P < 0.05) reduced Cronobacter to less than detectable limits (<1 log CFU ml(-1)) in skim milk at 5.0 kGy and milk with 1% fat content and greater at 6.0 kGy dose levels. The D-value for Cronobacter in TSB was significantly (P < 0.05) lower than those in milk samples. Treatment with X-rays could be an effective and safe alternative technology to control pathogenic bacteria (Cronobacter) in the dairy industry.

Internalization of Listeria monocytogenes in cantaloupes during dump tank washing and hydrocooling.

PubMed

Macarisin, Dumitru; Wooten, Anna; De Jesus, Antonio; Hur, Minji; Bae, Seonjae; Patel, Jitendra; Evans, Peter; Brown, Eric; Hammack, Thomas; Chen, Yi

2017-09-18

Recent listeriosis outbreaks and recalls associated with cantaloupes urge for studies to understand the mechanisms of cantaloupe contamination by Listeria monocytogenes. Postharvest practices such as washing and hydrocooling were suggested to facilitate the contamination of fresh fruits by human pathogens. This study assessed the potential of L. monocytogenes internalization into cantaloupes during dump tank washing and immersion-type hydrocooling in water contaminated with L. monocytogenes. The effect of cantaloupe cultivar, water temperature, and harvesting technique on L. monocytogenes internalization was also evaluated. Full slip (cantaloupe without any residual stem) Western and Eastern cultivar cantaloupes were pre-warmed to 42°C (to imitate peak-high field temperatures of freshly harvested cantaloupes) and then immersed in water at 6°C and 18°C containing 4 and 6logCFU/ml of L. monocytogenes. Clipped (cantaloupe with short stem residues obtained by clipping the stem at harvest) Western and Eastern cantaloupes were pre-warmed to 42°C and then immersed in water at 6°C containing 6logCFU/ml of L. monocytogenes. Additionally, full slip and clipped Western cantaloupes were equilibrated to 18°C and then immersed in water at 18°C containing 6logCFU/ml of L. monocytogenes (isothermal immersion without temperature differential). Water containing L. monocytogenes infiltrated both full slip and clipped cantaloupes through the stems/stem scars and was then distributed along the vascular system in hypodermal mesocarp reaching the calyx area of the fruit. The current study demonstrated that, under experimental conditions, L. monocytogenes can internalize into cantaloupes during immersion in water contaminated by L. monocytogenes, both in the presence and absence of temperature differential, and that temperature differential moderately enhanced the internalization of L. monocytogenes. The incidence and levels of L. monocytogenes internalized in the middle-mesocarp were significantly affected by harvesting technique but not by cantaloupe cultivar. Published by Elsevier B.V.

A novel quantitative reverse-transcription PCR (qRT-PCR) for the enumeration of total bacteria, using meat micro-flora as a model.

PubMed

Dolan, Anthony; Burgess, Catherine M; Barry, Thomas B; Fanning, Seamus; Duffy, Geraldine

2009-04-01

A sensitive quantitative reverse-transcription PCR (qRT-PCR) method was developed for enumeration of total bacteria. Using two sets of primers separately to target the ribonuclease-P (RNase P) RNA transcripts of gram positive and gram negative bacteria. Standard curves were generated using SYBR Green I kits for the LightCycler 2.0 instrument (Roche Diagnostics) to allow quantification of mixed microflora in liquid media. RNA standards were used and extracted from known cell equivalents and subsequently converted to cDNA for the construction of standard curves. The number of mixed bacteria in culture was determined by qRT-PCR, and the results correlated (r(2)=0.88, rsd=0.466) with the total viable count over the range from approx. Log(10) 3 to approx. Log(10) 7 CFU ml(-1). The rapid nature of this assay (8 h) and its potential as an alternative method to the standard plate count method to predict total viable counts and shelf life are discussed.

Inactivation of Escherichia coli, Listeria monocytogenes, and Salmonella Enteritidis by Cymbopogon citratus D.C. Stapf. Essential Oil in Pineapple Juice.

PubMed

Leite, Caroline Junqueira Barcellos; de Sousa, Jossana Pereira; Medeiros, José Alberto da Costa; da Conceição, Maria Lúcia; dos Santos Falcão-Silva, Vivyanne; de Souza, Evandro Leite

2016-02-01

In the present study, the efficacy of Cymbopogon citratus D.C. Stapf. essential oil (CCEO) to provoke a 5-log CFU/ml (5-log) inactivation in a mixed composite of Escherichia coli, Listeria monocytogenes, and Salmonella enterica serovar Enteritidis in pineapple (Ananas comosus (L.) Merril) juice (4°C) was assessed. Moreover, the effects of CCEO on the physicochemical and sensory quality parameters of pineapple juice were evaluated. The MIC of CCEO was 5 μl/ml against the composite mix examined. For L. monocytogenes and E. coli inoculated in juice containing CCEO (5, 2.5, and 1.25 μl/ml), a ≥5-log reduction was detected after 15 min of exposure. This same result was obtained for Salmonella Enteritidis incubated alone in pineapple juice containing CCEO at 5 and 2.5 μl/ml. Overall, Salmonella Enteritidis was the most tolerant and L. monocytogenes was the most sensitive to CCEO. The physicochemical properties (pH, titratable acidic [citric acid per 100 g], and soluble solids) of pineapple juice containing CCEO (2.5 and 1.25 μl/ml) were maintained. Juice containing CCEO (2.5 and 1.25 μl/ml) exhibited similar scores for odor, appearance, and viscosity compared with juice without CCEO. However, unsatisfactory changes in taste and aftertaste were observed in juices containing CCEO. These results suggest that CCEO could be used as an alternative antimicrobial compound to ensure the safety of pineapple juice, although CCEO at the tested concentrations negatively impacted its taste. Therefore, further studies are needed to determine the balance between microbial safety and taste acceptability of pineapple juice containing CCEO.

Occurrence of potentially pathogenic Vibrio in oysters (Crassostrea gigas) and waters from bivalve mollusk cultivations in the South Bay of Santa Catarina.

PubMed

Ramos, Roberta Juliano; Miotto, Letícia Adélia; Miotto, Marília; Silveira Junior, Nelson; Cirolini, Andréia; Silva, Helen Silvestre da; Rodrigues, Dália dos Prazeres; Vieira, Cleide Rosana Werneck

2014-01-01

This research aimed to identify and quantify potentially pathogenic Vibrio from different cultivations of bivalve shellfish in the State of Santa Catarina, Brazil, and water regions in the South Bay, as well as correlate the incidence of these microorganisms with the physicochemical parameters of marine waters. Between October 2008 and March 2009, 60 oyster and seawater samples were collected from six regions of bivalve mollusk cultivation, and these samples were submitted for Vibrio counts. Twenty-nine (48.3%) oyster samples were revealed to be contaminated with one or more Vibrio species. The Vibrio parahaemolyticus and Vibrio vulnificus counts in the samples ranged from < 0.5 log10 Most Probable Number (MPN) g(-1) to 2.3 log10 MPN g(-1) oyster and from < 0.5 log10 MPN g(-1) to 2.1 log10 MPN g(-1) oyster, respectively. Of the 60 seawater samples analyzed, 44 (73.3%) showed signs of contamination with one or more vibrio species. The counts of V. parahaemolyticus and V. vulnificus in the samples ranged from < 0.3 log10 MPN·100mL(-1) to 1.7 log10MPN·100mL(-1) seawater and from < 0.3 log10 MPN·100mL(-1) to 2.0 log10 MPN·100mL(-1) seawater, respectively. A positive correlation between V. vulnificus counts and the seawater temperature as well as a negative correlation between the V. parahaemolyticus counts and salinity were observed. The results suggest the need to implement strategies to prevent vibrio diseases from being transmitted by the consumption of contaminated bivalve shellfish.

Microbial communities and fecal indicator bacteria associated with Cladophora mats on beach sites along Lake Michigan shores.

PubMed

Olapade, Ola A; Depas, Morgan M; Jensen, Erika T; McLellan, Sandra L

2006-03-01

A high biomasses of Cladophora, a filamentous green alga, is found mainly during the summer along the shores of Lake Michigan. In this study, the abundance and persistence of the fecal indicator bacterium Escherichia coli and sulfate-reducing bacteria (SRB) on Cladophora mats collected at Lake Michigan beaches were evaluated using both culture-based and molecular analyses. Additionally, 16S rRNA gene cloning and sequencing were used to examine the bacterial community composition. Overall, E. coli was detected in all 63 samples obtained from 11 sites, and the average levels at most beaches ranged from 2,700 CFU/100 g (wet weight) of Cladophora to 7,500 CFU/100 g of Cladophora. However, three beaches were found to have site average E. coli densities of 12,800, 21,130, and 27,950 CFU/100 g of Cladophora. The E. coli levels in the lake water collected at the same time from these three sites were less than the recommended U.S. Environmental Protection Agency limit, 235 CFU/100 ml. E. coli also persisted on Cladophora mats in microcosms at room temperature for more than 7 days, and in some experiments it persisted for as long as 28 days. The SRB densities on Cladophora mats were relatively high, ranging from 4.4x10(6) cells/g (6.64 log CFU/g) to 5.73x10(6) cells/g (6.76 log CFU/g) and accounting for between 20% and 27% of the total bacterial counts. Partial sequences of the 16S rRNA gene clones revealed a phylogenetically diverse community, in which the Cytophaga-Flavobacterium-Bacteroides cluster and the low-G+C-content gram-positive bacteria were the dominant organisms, accounting for 40% and 12.8%, respectively, of the total clone library. These results further reveal the potential public health and ecological significance of Cladophora mats that are commonly found along the shoreline of Lake Michigan, especially with regard to the potential to harbor microorganisms associated with fecal pollution and odor-causing bacteria.

Fosfomycin synergy in vitro with amoxicillin, daptomycin, and linezolid against vancomycin-resistant Enterococcus faecium from renal transplant patients with infected urinary stents.

PubMed

Descourouez, Jillian L; Jorgenson, Margaret R; Wergin, Justine E; Rose, Warren E

2013-03-01

Fosfomycin is a potential option for vancomycin-resistant enterococcus (VRE) infections despite limited in vitro and clinical data. In this study, 32 VRE isolates from renal transplant patients with urinary stent infections were susceptible to fosfomycin, daptomycin, and linezolid and resistant to amoxicillin, minocycline, and nitrofurantoin based on their MIC(50)s and MIC(90)s. Fosfomycin was bacteriostatic at 0.5 to 16× the MIC (32 to 2,048 μg/ml); synergy occurred when fosfomycin was combined with daptomycin (2.8 to 3.9 log(10) CFU/ml kill; P < 0.001) or amoxicillin (2.6 to 3.4; P < 0.05). These combinations may be potent options to treat VRE urinary infections pending investigation of clinical efficacy.

Validation of a Clostridium Endospore Viability Assay and Analysis of Greenland Ices and Atacama Desert Soils▿ †

PubMed Central

Yang, Wan-Wan; Ponce, Adrian

2011-01-01

A microscopy-based endospore viability assay (micro-EVA) capable of enumerating germinable Clostridium endospores (GCEs) in less than 30 min has been validated and employed to determine GCE concentrations in Greenland ices and Atacama Desert soils. Inoculation onto agarose doped with Tb3+ and d-alanine triggers Clostridium spore germination and the concomitant release of ∼108 molecules of dipicolinic acid (DPA) per endospore, which, under pulsed UV excitation, enables enumeration of resultant green Tb3+-DPA luminescent spots as GCEs with time-gated luminescence microscopy. The intensity time courses of the luminescent spots were characteristic of stage I Clostridium spore germination dynamics. Micro-EVA was validated against traditional CFU cultivation from 0 to 1,000 total endospores/ml (i.e., phase-bright bodies/ml), yielding 56.4% ± 1.5% GCEs and 43.0% ± 1.0% CFU. We also show that d-alanine serves as a Clostridium-specific germinant (three species tested) that inhibits Bacillus germination of spores (five species tested) in that endospore concentration regime. Finally, GCE concentrations in Greenland ice cores and Atacama Desert soils were determined with micro-EVA, yielding 1 to 2 GCEs/ml of Greenland ice (versus <1 CFU/ml after 6 months of incubation) and 66 to 157 GCEs/g of Atacama Desert soil (versus 40 CFU/g soil). PMID:21296951

Effect of Lactobacillus plantarum isolated from digestive tract of wild shrimp on growth and survival of white shrimp (Litopenaeus vannamei) challenged with Vibrio harveyi.

PubMed

Kongnum, Khanitta; Hongpattarakere, Tipparat

2012-01-01

Two hundred and two strains of lactic acid bacteria (LAB) isolated from digestive tracts of cultivated and wild adult shrimp, including Litopenaeus vannamei, Metapenaeus brevicornis and Penaeus merguiensis were selected based on their antibacterial activity against Vibrio harveyi. LAB strain of MRO3.12 exhibiting highest reduction of V. harveyi was identified as Lactobacillus plantarum MRO3.12 based on the nucleotide sequence of its 16S rDNA, which showed 99% (780/786 bp) homology to L. plantarum strain L5 (GenBank accession number DQ 239698.1). Co-cultivation of V. harveyi and L. plantarum MRO3.12 showed complete reduction of V. harveyi at 24 h under aerobic and anaerobic conditions, whereas L. plantarum increased from 5.29 to 9.47 log CFU ml(-1). After 6-week feeding trial with L. plantarum supplemented diet, white shrimp (L. vannamei) exhibited significant differences (p < 0.05) in relative growth rate (% RGR), feed conversion ratio (FCR) and survival compared to the control group fed with non-supplemented diet. LAB-fed group showed 98.89% survival, whereas only 68.89% survival was observed in the control group. LAB from the digestive tract of probiotic-fed shrimp showed higher level of 5.0 ± 0.14 log CFU/g than the non-supplemented ones (3.34 ± 0.21 log CFU/g). However, total bacterial and non-fermenting vibrios counts decreased in shrimps fed on L. plantarum. Ten days after infection with V. harveyi (5.3-5.5 log CFU ml(-1)), significant survival (p < 0.05) of 77% was observed in LAB supplemented shrimp, while only 67% survival was observed in the control. Copyright © 2011 Elsevier Ltd. All rights reserved.

Use of gamma radiation for inactivating Salmonella spp., Escherichia coli O157:H7 and Listeria monocytogenes in tahini halva.

PubMed

Osaili, Tareq M; Al-Nabulsi, Anas A; Aljaafreh, Taqwa F

2018-08-02

Tahini halva is a traditional sweet product that is consumed with bread in different countries. It is a low water activity (a w ) product basically made by mixing and cooking tahini, sugar, citric acid and Saponaria officinalis root extract together. Tahini halva maybe contaminated with foodborne pathogens during any stage of production from tahini and other raw ingredients, workers, environment or contact surfaces. The objectives of the study were to i) investigate the efficacy of gamma radiation to inactivate Salmonella spp., Escherichia coli O157:H7 and Listeria monocytogenes in tahini halva, ii) evaluate the effect of pre-irradiation storage (0, 7 and 30 days at 21 °C) of tahini halva on the sensitivity of these microorganisms toward gamma radiation, and iii) evaluate the effect of post-irradiation storage of tahini halva for up to 6 months on the their survival characteristics. Tahini halva samples were inoculated with Salmonella spp., E. coli O157:H7 and L. monocytogenes separately then stored at 21 °C for 0, 7 and 30 days prior to irradiation at 0-4 KGy and for up to 6 months after irradiation at 4 KGy. Salmonella spp. were the most irradiation resistance among the tested microorganisms. Irradiation (0.8-4.0 KGy) reduced the bacteria in samples stored for 0, 7 and 30 days pre-irradiation in the range of 0.43-2.11, 0.45-2.68 and 0.52-2.7 log 10  CFU/g for Salmonella spp., 0.55-3.08, 0.66-3.00 and 0.60-2.80 log 10  CFU/g for E. coli O157:H7, and 0.69-2.96, 0.86-4.30, 0.62-3.29 log 10  CFU/g for L. monocytogenes, respectively. The D 10 -value, the irradiation dose needed to inactivate 1 log 10 of pathogen, was 1.83, 1.47 and 1.50 KGy for Salmonella spp., 1.28, 1.32 and 1.48 KGy for E. coli O157:H7, and 1.33, 0.94 and 1.27 KGy for L. monocytogenes in pre-irradiation stored samples for 0, 7 and 30 days, respectively. Post-irradiation storage was efficient in decreasing the levels of the microorganisms ca. ≥2 log 10  CFU/g in the first month and to undetected level after the second month of storage but enrichment results showed that Salmonella spp. and L. monocytogenes were detected in the samples until of the end of storage period. The study demonstrates that gamma radiation can be applied to inactivate of foodborne pathogens in tahini halva. Irradiation dose at 4 KGy can reduce Salmonella spp., E. coli O157:H7 and L. monocytogenes in tahini halva by 2-3 log 10  CFU/g. Storage of tahini halva before or after irradiation may reduce the risk of foodborne pathogens in the product. Copyright © 2018 Elsevier B.V. All rights reserved.

Evaluation of Ceftolozane-Tazobactam in Combination with Meropenem against Pseudomonas aeruginosa Sequence Type 175 in a Hollow-Fiber Infection Model.

PubMed

Montero, M; VanScoy, Brian D; López-Causapé, Carla; Conde, Haley; Adams, Jonathan; Segura, Concepción; Zamorano, Laura; Oliver, Antonio; Horcajada, Juan P; Ambrose, Paul G

2018-05-01

The aim of this study was to investigate the efficacy of ceftolozane-tazobactam in combination with meropenem against an extensively drug-resistant (XDR) Pseudomonas aeruginosa high-risk clone, sequence type 175, isolated in a Spanish university hospital. A 14-day hollow-fiber infection model was used to simulate clinical exposure of the two drug regimens alone and in combination, and serial samples were collected to determine drug concentrations and CFU counts. The untreated control failed, as did each study regimen when administered alone. However, when ceftolozane-tazobactam was administered in combination with meropenem, there was a >4-log 10 CFU/ml bacterial density reduction and suppression of resistance for the duration of the study. These data suggest that ceftolozane-tazobactam plus meropenem may be a useful combination for treating XDR P. aeruginosa . Copyright © 2018 American Society for Microbiology.

Bacillus sp. Acting as Dual Role for Corrosion Induction and Corrosion Inhibition with Carbon Steel (CS)

PubMed Central

Karn, Santosh K.; Fang, Guan; Duan, Jizhou

2017-01-01

Present work investigated the role of five different bacteria species as a corrosion inducer as well as corrosion inhibitor with carbon steel (CS). We observed the ability of different bacteria species on the metal surface attachment, biofilm formation, and determined Peroxidase, Catalase enzyme activity in the detached biofilm from the CS surface. We found that each strain has diverse conduct for surface attachment like DS1 3.3, DS2 2.5, DS3 4.3, DS4 4.0, and DS5 4.71 log cfu/cm2 and for biofilm 8.3 log cfu/cm2. The enzyme Peroxidase, Catalase was found in huge concentration inside the biofilm Peroxidase was maximum for DS4 36.0 U/ml and least for DS3 19.54 U/ml. Whereas, Catalase was highest for DS4, DS5 70.14 U/ml and least 57.2 U/ml for DS2. Scanning electron microscopy (SEM) was conducted to examine the biofilm and electrochemical impedance spectroscopy (EIS) were utilized to observe corrosion in the presence of bacteria. The electrochemical results confirmed that DS1, DS3, DS4, and DS5 strains have statistically significant MIC-factors (Microbially Influenced Corrosion) of 5.46, 8.51, 2.36, and 1.04, while DS2 protective effect factor of 0.89. Weight reduction results with carbon steel likewise supports that corrosion was initiated by DS1 and DS3, while DS2 and DS5 have no any impact though with DS4 we watched less weight reduction however assumed no role in the corrosion. We established the relation of Peroxidase enzyme activity of the isolates. DS1, DS3 and having Peroxidase in the range 22.18, 19.54 U/ml which induce the corrosion whereas DS2 and DS5 having 28.57 and 27.0 U/ml has no any effect and DS4 36 U/ml has inhibitory effect, increasing concentration inhibiting the corrosion. For Catalase DS1, DS3 have 67.28, 61.57 U/ml which induce corrosion while DS2 and DS5 57.71 and 59.14 U/ml also has no effect whereas DS4 70.14 U/ml can inhibit corrosion. Results clearly express that in a specific range both enzymes can induce the corrosion. Our goals are to pursuit and locate the potential role of the enzyme in corrosion induction and inhibition. There is still further work is proceeded for the more profound perception. PMID:29114242

Antimicrobial blue light inactivation of biofilms formed by clinical isolates of multidrug-resistant microorganisms

NASA Astrophysics Data System (ADS)

Ferrer-Espada, Raquel; Fang, Yanyan; Dai, Tianhong

2018-02-01

Antibiotic resistance is one of the most serious threats to public health. It is estimated that at least 23,000 people die each year in the USA as a direct result of antibiotic-resistant infections. In addition, many antibiotic-resistant microorganisms develop biofilms, surface-associated microbial communities that are extremely resistant to antibiotics and the immune system. A light-based approach, antimicrobial blue light (aBL), has attracted increasing attention due to its intrinsic antimicrobial effect without the involvement of exogenous photosensitizers. In this study, we investigated the effectiveness of this non-antibiotic approach against biofilms formed by multidrug-resistant (MDR) microorganisms. MDR Acinetobacter baumannii, Escherichia coli, Candida albicans, and Pseudomonas aeruginosa biofilms were grown either in 96-well microtiter plates for 24 h or in a CDC biofilm reactor for 48 h, and then exposed to aBL at 405 nm emitted from a light-emitting diode (LED). We demonstrated that, for the biofilms grown in the CDC biofilm reactor, approximately 1.88 log10 CFU reduction was achieved in A. baumannii, 2.78 log10 CFU in E. coli and 3.18 log10 CFU in P. aeruginosa after 162 J/cm2 , 576 J/cm2 and 500 J/cm2 aBL were delivered, respectively. For the biofilms formed in the 96-well microtiter plates, 5.67 and 2.46 log10 CFU reduction was observed in P. aeruginosa and C. albicans polymicrobial biofilm after an exposure of 216 J/cm2 . In conclusion, aBL is potentially an alternative non-antibiotic approach against MDR biofilm-related infections. Future studies are warranted to investigate other important MDR microorganisms, the mechanism of action of aBL, and aBL efficacy in vivo.

Efficacy of feed additives against Campylobacter in live broilers during the entire rearing period.

PubMed

Guyard-Nicodème, M; Keita, A; Quesne, S; Amelot, M; Poezevara, T; Le Berre, B; Sánchez, J; Vesseur, P; Martín, Á; Medel, P; Chemaly, Marianne

2016-02-01

Poultry meat is the major source of human campylobacteriosis, the most frequently reported zoonosis in the EU. The prevalence of Campylobacter colonization in European broiler flocks is 71%. Despite considerable efforts, there is still no effective strategy available to prevent or reduce Campylobacter colonization in broilers. This study tested a wide variety of feed additives to reduce Campylobacter shedding in primary poultry production. Twelve additives containing organic or fatty acids, monoglycerides, plant extracts, prebiotics, or probiotics were tested. For each additive, broilers contaminated with Campylobacter jejuni were fed with an additive free diet (control group) or with a supplemented diet (treated group) and Campylobacter loads compared at three sampling times. No treatment was able to prevent broiler colonization by Campylobacter, and there was a high degree of variation in contamination among the birds. At 14 d of age, eight treatments significantly decreased the colonization level compared to the control group by a maximum of 2 log10 CFU/g. At 35 d of age, three of these treatments still had a significant effect with a maximum reduction of 1.88 log10 CFU/g for a probiotic. At 42 d of age, only one short-chain fatty acid was still significantly efficient with a mean reduction over 2 log10 CFU/g. In addition, a probiotic and a prebiotic-like compound significantly decreased the contamination by a maximum of 3 log10 CFU/g, only at the 42-d sampling period. This study gives promising results regarding the use of feed additives to reduce Campylobacter infection in flocks. Nevertheless, a global approach, combining intervention measures at the different steps of the broiler meat production chain could have a greater impact on the reduction of public health risk. © 2015 Poultry Science Association Inc.

Decreased dosage of acidified sodium chlorite reduces microbial contamination and maintains organoleptic qualities of ground beef products.

PubMed

Bosilevac, Joseph M; Shackelford, Steven D; Fahle, Rick; Biela, Timothy; Koohmaraie, Mohammad

2004-10-01

Acidified sodium chlorite (ASC) spray was evaluated at decreased dosages and application rates to determine its efficacy for reducing bacterial contamination on boneless beef trimmings used for production of raw ground beef products while maintaining desirable consumer qualities in the finished ground beef products. Two different applications of ASC (600 ppm applied at a rate of 1.3 oz/lb and 300 ppm applied at a rate of 1 oz/lb) were used to treat boneless beef trimmings before grinding. The effect of ASC treatment on 50/50 lean beef trimmings was greater than on 90/10 trimmings. ASC at 600 ppm reduced both the aerobic plate counts (APC) and Enterobacteriaceae counts (EBC) by 2.3 log CFU/g on 50/50 trimmings, whereas treatment with 300 ppm ASC reduced APC and EBC of 50/50 trimmings by 1.1 and 0.7 log CFU/g, respectively. Ground beef formulations of 90/10 and 73/27 were produced from the treated boneless beef trim and packaged in chubs and in modified atmosphere packaging. The efficacy of ASC spray treatment to inhibit APC and EBC over the shelf life of each ground beef product was monitored. The APC and EBC in ground beef chubs were reduced by 1.0 to 1.5 log CFU/g until day 20. The APC and EBC for products in modified atmosphere packaging were reduced 1.5 to 3.0 log CFU/g throughout their shelf life. Both decreased dosages of ASC were equally effective on 90/10 lean ground beef, but the 300 ppm ASC treatment was slightly better at reducing the EBC of 73/27 ground beef. The organoleptic qualities (color, odor, and taste) of the ground beef products treated with 300 ppm ASC were found to be superior to those treated with 600 ppm ASC. Our results indicated that decreased dosages of ASC reduce contamination and lengthen the shelf life of ground beef. Furthermore, the 300 ppm ASC treatment reduced bacterial counts while maintaining desirable organoleptic ground beef qualities.

Magnetic nanoparticle-enhanced PCR for the detection and identification of Staphylococcus aureus and Salmonella enteritidis.

PubMed

Houhoula, Dimitra; Papaparaskevas, Joseph; Zatsou, Katerina; Nikolaras, Nikolaos; Malkawi, Hanan I; Mingenot-Leclercq, Marie-Paule; Konteles, Spyros; Koussisis, Stamatis; Tsakris, Athanassios; Charvalos, Ekatherina

2017-07-01

This paper evaluated magnetic nanoparticle-enhanced PCR for the detection and identification of Staphylococcus aureus and Salmonella enteritidis. Two different types of magnetic nanoparticles designated MPIO (iron concentration 2.5 mg/ml, size 1 µm) and NP (iron concentration 8.7 mg/ml, size 60 nm), both conjugated with S. aureus or S. enteritidis antibodies were evaluated as an enrichment procedure for PCR-detection of the pathogens in Trypticase Soy Broth, milk, blood and meat broth. Bacterial suspensions (1.5x108 cfu/ml) were prepared and serial diluted 10-1. The MPIO and NP nanoparticles were added, followed by incubation for 1 hour at room temperature, magnetic separation of the pellet, DNA extraction and PCR, targeting the femA and invA sequences. The nanoparticle-free and the NP-supplemented dilutions were positive down to the 1.5x102 cfu/ml concentration for both bacteria. The MPIO-supplemented dilutions were positive down to approx. 2x100 cfu/ml concentration, respectively. Bacteria-free TSB was negative by PCR. MPIO nanoparticles (size 1 µm) enhanced the detection of S. aureus and S. enteritidis by PCR, whilst NP nanoparticles (size 60 nm) did not, thus indicating that the size of the magnetic nanoparticles play a significant role in the enrichment procedure.

Growth of Listeria spp. in shredded cabbage is enhanced by a mild heat treatment.

PubMed

Ells, Timothy C; Truelstrup Hansen, Lisbeth

2010-03-01

Mild thermal processing can enhance the shelf life of cut fruits and vegetables by delaying the onset of spoilage and preserving the organoleptic properties of shredded cabbage. However, food safety issues related to this process have not been fully investigated. Therefore, the survival and growth of Listeria spp. on cabbage treated in this manner was examined. Experimentally, 24 strains of Listeria spp. (including L. monocytogenes) were inoculated onto cut and intact cabbage tissues and stored at 5 degrees C. All strains on intact tissues exhibited a moderate decline in numbers (up to 1.0 log CFU/cm(2)) over the 28-day storage period. Conversely, cut tissue supported growth of most strains during the first 7 to 14 days of incubation with maximum increases of 1.2 log CFU/cm(2). Subsequently, the survival or growth on heat-treated (50 degrees C for 3 min) and untreated shredded cabbage of four L. monocytogenes and four nonpathogenic Listeria spp. strains were compared during storage for 21 days at 5 degrees C. Growth on untreated shred for all strains was similar to the results observed on cut tissue with a maximum increase of approximately 1.0 log CFU/g. However, in the heat-treated cabbage shred all strains displayed a rapid increase in growth (up to 2.5 log CFU/g) during the first 7 days of incubation, which may be indicative of the destruction of an endogenous growth-inhibiting compound within the cabbage. In conclusion, this study shows that mild thermal treatments of cut cabbage may promote pathogen growth if other inimical barriers are not implemented downstream of the thermal treatment.

Salmonella and Escherichia coli O157:H7 Inactivation, Color, and Bioactive Compounds Enhancement on Raspberries during Frozen Storage after Decontamination Using New Formula Sanitizer Washing or Pulsed Light.

PubMed

Xu, Wenqing; Chen, Haiqiang; Wu, Changqing

2016-07-01

Berries are normally washed before they are frozen. Washing with sanitizer and treatment with pulsed light (PL) were studied for their effectiveness to inactivate foodborne pathogens on raspberries during frozen storage, while maintaining or enhancing major quality parameters. Raspberries were inoculated with Salmonella or Escherichia coli O157:H7 and then underwent a washing treatment with citric acid plus sodium dodecyl sulfate (CA+SDS) or citric acid plus thymol (CA+THY) or treatment with PL (dry PL, water-assisted [wet] PL, and PL-SDS). Pathogen survival was determined immediately after treatments and during frozen storage at -20°C for 3 months. Washing with CA+SDS or CA+THY significantly reduced Salmonella (by 3.6 and 3.2 log CFU/g, respectively) and E. coli O157:H7 (by 4.1 and 3.7 log CFU/g, respectively). At the end of storage, washing with CA+SDS reduced Salmonella to 0.6 log CFU/g and E. coli O157:H7 to 0.5 log CFU/g; washing with CA+THY reduced Salmonella to 0.9 log CFU/g and E. coli O157:H7 to 0.5 log CFU/g. PL-SDS showed decontamination efficacy on raspberries, with 0.7 log CFU/g Salmonella and 0.9 log CFU/g E. coli O157:H7 surviving at the end of storage; in comparison, in the control, 1.6 log CFU/g Salmonella and 1.5 log CFU/g E. coli O157:H7 survived. Pathogen survival in raspberries that had been washed or treated with PL-SDS was significantly lower than in untreated raspberries. Major quality parameters, including color, total phenolic content, total anthocyanin content, total bacterial count, and total yeast and mold counts, were evaluated on raspberries immediately after treatments and during frozen storage. Redness increased in PL-treated raspberries. At the end of storage, PL-treated raspberries had significantly higher total phenolic content and total anthocyanin content compared with control samples. Washing with sanitizers and treatment with PL decreased the total bacterial count and total yeast and mold counts on raspberries and maintained the low counts. Our findings suggest that washing with a sanitizer or treatment with PL could be used to process frozen raspberries for enhanced food safety and quality.

Simultaneous detection of Escherichia coli O157:H7 and Salmonella Typhimurium using quantum dots as fluorescence labels.

PubMed

Yang, Liju; Li, Yanbin

2006-03-01

In this study, we explored the use of semiconductor quantum dots (QDs) as fluorescence labels in immunoassays for simultaneous detection of two species of foodborne pathogenic bacteria, Escherichia coli O157:H7 and Salmonella Typhimurium. QDs with different sizes can be excited with a single wavelength of light, resulting in different emission peaks that can be measured simultaneously. Highly fluorescent semiconductor quantum dots with different emission wavelengths (525 nm and 705 nm) were conjugated to anti-E. coli O157 and anti-Salmonella antibodies, respectively. Target bacteria were separated from samples by using specific antibody coated magnetic beads. The bead-cell complexes reacted with QD-antibody conjugates to form bead-cell-QD complexes. Fluorescent microscopic images of QD labeled E. coli and Salmonella cells demonstrated that QD-antibody conjugates could evenly and completely attach to the surface of bacterial cells, indicating that the conjugated QD molecules still retain their effective fluorescence, while the conjugated antibody molecules remain active and are able to recognize their specific target bacteria in a complex mixture. The intensities of fluorescence emission peaks at 525 nm and 705 nm of the final complexes were measured for quantitative detection of E. coli O157:H7 and S. Typhimurium simultaneously. The fluorescence intensity (FI) as a function of cell number (N) was found for Salmonella and E. coli, respectively. The regression models can be expressed as: FI = 60.6 log N- 250.9 with R(2) = 0.97 for S. Typhimurium, and FI = 77.8 log N- 245.2 with R(2) = 0.91 for E. coli O157:H7 in the range of cell numbers from 10(4) to 10(7) cfu ml(-1). The detection limit of this method was 10(4) cfu ml(-1). The detection could be completed within 2 hours. The principle of this method could be extended to detect multiple species of bacteria (3-4 species) simultaneously, depending on the availability of each type of QD-antibody conjugates with a unique emission peak and the antibody coated magnetic beads specific to each species of bacteria.

Efficacy of cetylpyridinium chloride against Listeria monocytogenes and its influence on color and texture of cooked roast beef.

PubMed

Singh, M; Thippareddi, H; Phebus, R K; Marsden, J L; Herald, T J; Nutsch, A L

2005-11-01

Sliced (cut) and exterior (intact) surfaces of restructured cooked roast beef were inoculated with Listeria monocytogenes, treated with cetylpyridinium chloride (CPC; immersion in 500 ml of 1% solution for 1 min), individually vacuum packaged, and stored for 42 days at 0 or 4 degrees C. Noninoculated samples were similarly treated, packaged, and stored to determine effects on quality (color and firmness) and on naturally occurring bacterial populations, including aerobic plate counts and lactic acid bacteria. Immediately after CPC treatment, regardless of inoculation level, L. monocytogenes populations were reduced (P = 0.05) by about 2 log CFU/cm2 on sliced surfaces and by about 4 log CFU/cm2 on exterior surfaces. Throughout 42 days of refrigerated storage (at both 0 and 4 degrees C), L. monocytogenes populations on CPC-treated samples remained lower (P = 0.05) than those of nontreated samples for both surface types. After 42 days of storage at both 0 and 4 degrees C, aerobic plate count and lactic acid bacteria populations of treated samples were 1 to 1.5 log CFU/cm2 lower (P = 0.05) than those of nontreated samples for both surface types. CPC treatment resulted in negligible effects (P > 0.05) on the color (L*, a*, and b* values) of exterior and sliced roast beef surfaces during storage. For both sliced and exterior surfaces, CPC-treated samples were generally less firm than nontreated samples. CPC treatment effectively reduced L. monocytogenes populations on roast beef surfaces and resulted in relatively minor impacts on color and texture attributes. CPC treatment, especially when applied to products prior to slicing, may serve as an effective antimicrobial intervention for ready-to-eat meat products.

Performance evaluation of the QIAGEN EZ1 DSP Virus Kit with Abbott RealTime HIV-1, HBV and HCV assays.

PubMed

Schneider, George J; Kuper, Kevin G; Abravaya, Klara; Mullen, Carolyn R; Schmidt, Marion; Bunse-Grassmann, Astrid; Sprenger-Haussels, Markus

2009-04-01

Automated sample preparation systems must meet the demands of routine diagnostics laboratories with regard to performance characteristics and compatibility with downstream assays. In this study, the performance of QIAGEN EZ1 DSP Virus Kit on the BioRobot EZ1 DSP was evaluated in combination with the Abbott RealTime HIV-1, HCV, and HBV assays, followed by thermalcycling and detection on the Abbott m2000rt platform. The following performance characteristics were evaluated: linear range and precision, sensitivity, cross-contamination, effects of interfering substances and correlation. Linearity was observed within the tested ranges (for HIV-1: 2.0-6.0 log copies/ml, HCV: 1.3-6.9 log IU/ml, HBV: 1.6-7.6 log copies/ml). Excellent precision was obtained (inter-assay standard deviation for HIV-1: 0.06-0.17 log copies/ml (>2.17 log copies/ml), HCV: 0.05-0.11 log IU/ml (>2.09 log IU/ml), HBV: 0.03-0.07 log copies/ml (>2.55 log copies/ml)), with good sensitivity (95% hit rates for HIV-1: 50 copies/ml, HCV: 12.5 IU/ml, HBV: 10 IU/ml). No cross-contamination was observed, as well as no negative impact of elevated levels of various interfering substances. In addition, HCV and HBV viral load measurements after BioRobot EZ1 DSP extraction correlated well with those obtained after Abbott m2000sp extraction. This evaluation demonstrates that the QIAGEN EZ1 DSP Virus Kit provides an attractive solution for fully automated, low throughput sample preparation for use with the Abbott RealTime HIV-1, HCV, and HBV assays.

Treatment of Candida albicans biofilms with low-temperature plasma induced by dielectric barrier discharge and atmospheric pressure plasma jet

NASA Astrophysics Data System (ADS)

Koban, Ina; Matthes, Rutger; Hübner, Nils-Olaf; Welk, Alexander; Meisel, Peter; Holtfreter, Birte; Sietmann, Rabea; Kindel, Eckhard; Weltmann, Klaus-Dieter; Kramer, Axel; Kocher, Thomas

2010-07-01

Because of some disadvantages of chemical disinfection in dental practice (especially denture cleaning), we investigated the effects of physical methods on Candida albicans biofilms. For this purpose, the antifungal efficacy of three different low-temperature plasma devices (an atmospheric pressure plasma jet and two different dielectric barrier discharges (DBDs)) on Candida albicans biofilms grown on titanium discs in vitro was investigated. As positive treatment controls, we used 0.1% chlorhexidine digluconate (CHX) and 0.6% sodium hypochlorite (NaOCl). The corresponding gas streams without plasma ignition served as negative treatment controls. The efficacy of the plasma treatment was determined evaluating the number of colony-forming units (CFU) recovered from titanium discs. The plasma treatment reduced the CFU significantly compared to chemical disinfectants. While 10 min CHX or NaOCl exposure led to a CFU log10 reduction factor of 1.5, the log10 reduction factor of DBD plasma was up to 5. In conclusion, the use of low-temperature plasma is a promising physical alternative to chemical antiseptics for dental practice.

[Development and comparative evaluation of up-converting phosphor technology based lateral flow assay for rapid detection of Yersinia pestis, Bacillus anthracis spore and Brucella spp].

PubMed

Li, Chunfeng; Zhang, Pingping; Wang, Xiaoying; Liu, Xiao; Zhao, Yong; Sun, Chongyun; Wang, Chengbin; Yang, Ruifu; Zhou, Lei

2015-01-01

To develop an up-converting phosphor technology based lateral flow (UPT-LF) assay for rapid and quantitative detection of Yersinia pestis, Bacillus anthracis spore and Brucella spp.and make the comparison with BioThreat Alert (BTA) test strips (Tetracore Inc., USA). Using up-converting phosphor nano-particles (UCP-NPs) as the bio-marker, three double-antibody-sandwich model based UPT-LF strips including Plague-UPT-LF, Anthrax-UPT-LF, Brucella-UPT-LF were prepared and its sensitivity, accuracy, linearity and specificity were determined by detecting 10(10), 10(9), 10(8), 10(7), 10(6), 10(5) and 0 CFU/ml series of concentrations of Y.pestis, B.anthracis, Brucella standards and other 27 kinds of 10(9) CFU/ml series of contrations of bacteria strains.Furthermore, the speed, sensitivity and accuracy of bacteria standards and simulated sample detection were compared between UPT-LF and BTA system. The detection limit of Plague-UPT-LF, Anthrax-UPT-LF and Brucella-LF was 10(5) CFU/ml. The CV of series of bacteria concentrations was ≤ 15%, and the r between lg (T/C-cut-off) and lg (concentration) was 0.996,0.998 and 0.999 (F values were 1 647.57, 743.51 and 1 822.17. All the P values were <0.001), respectively. The specificity of Plague-UPT-LF and Brucella-LF were excellent, while that of Anthrax-UPT-LF was a little bit regretful because of non-specific reaction with two isolates of B. subtilis and one B.cereus. On-site evaluation showed the detection time of UPT-LF for all Y.pestis, B.anthracis spore and Brucella spp.was 33, 36 and 37 min, while BTA was 115, 115 and 111 min, which revealed the higher detection speed and sensitivity of UPT-LF comparing with BTA. The negative rate of two methods for blank standard was both 5/5, the sensitivity of UPT-LF for Y.pestis,B.anthracis spore and Brucella spp. was all 10(5) CFU/ml, then BTA was 10(6), 10(6) and 10(5) CFU/ml, respectively. The detection rate of UPT-LF for all three bacteria analog positive samples was 16/16, while BTA for B.anthracis was 7/16 only. The good performance including rapidness, simplicity and high sensitivity will bring the bright future of UPT-LF to be broadly used on-site as first response to bio-terrorism.

An Assessment of Coliform Bacteria in Water Sources Near Appalachian Trail Shelters Within the Great Smoky Mountains National Park.

PubMed

Reed, Brian C; Rasnake, Mark S

2016-03-01

Hikers and campers are exposed to risks while in the wilderness. One of these risks is the possibility of contracting an illness, including infectious diarrhea. This project tested for coliform bacteria in water samples taken near popular Appalachian Trail shelters. Water was collected from access points within the Great Smoky Mountains National Park. Samples were collected in sterile bottles and inoculated on a commercially available coliform detection kit for quantitative determination of total coliform and Escherichia coli counts. Water samples were taken during summer and fall seasons. During summer, 7 of 10 samples were positive for coliform bacteria and 6 of those 7 for E coli. The most probable number (MPN) of colony-forming units (CFU) for coliform bacteria ranged from 0 to 489 CFU/100 mL, with the MPN for E coli varying from 0 to 123 CFU/100 mL. These data differed from the fall collection, revealing 3 of 7 samples positive for coliform bacteria and 1 of those 3 for E coli. The MPN of CFU for coliform bacteria in fall samples varied from 0 to 119 CFU/100 mL and 0 to 5 to CFU/100 mL for E coli. Environmental Protection Agency drinking water standards set the standard of 0 CFU/100 mL to be considered safe. This analysis of water samples along the Appalachian Trail emphasizes that the majority of water access points require treatment during the summer season. Coliform burden was not as high through the fall months. These data suggest one infectious disease risk for wilderness travelers. Copyright © 2016 Wilderness Medical Society. Published by Elsevier Inc. All rights reserved.

Incorporation of preservatives in polylactic acid films for inactivating Escherichia coli O157:H7 and extending microbiological shelf life of strawberry puree.

PubMed

Jin, Tony; Zhang, Howard; Boyd, Glenn

2010-05-01

Antimicrobial films of polylactic acid polymer incorporated with nisin, EDTA, sodium benzoate (SB), potassium sorbate (PS), and their combinations were developed, and their antimicrobial effects on the inactivation of Escherichia coli O157:H7 and natural background microflora (total aerobic bacteria, molds, and yeasts) in strawberry puree at 10 and 22 degrees C were determined. Direct addition of SB+PS to strawberry puree was also used as a comparison with SB+PS film treatment. The combination treatment reduced the cell populations of E. coli O157:H7 from 3.5 log CFU/ml to undetectable levels (<1 CFU/ml) after 14 days and 1 day at 10 and 22 degrees C, respectively, while the cells of E. coli O157:H7 in control samples survived up to 48 days at 10 degrees C and more than 14 days at 22 degrees C. The SB+PS film treatment produced a greater reduction of population of E. coli O157:H7 cells than did the SB+PS direct addition treatment. Similar results were observed for inactivation of natural microflora. In general, the antimicrobial effect was in the following order: film combination > SB+PS film > SB+PS direct addition > EDTA film > nisin film. The data obtained in this study suggest two approaches toward the development of control interventions against E. coli O157:H7 and extension of the microbiological shelf life of strawberry puree: (i) using antimicrobial packaging and (ii) using combinations of preservatives. The film formulas developed here can be used to make bottles or as coatings on the surface of bottles for use in liquid food packaging.

Fasting hypochlorhydria with gram positive gastric flora is highly prevalent in healthy old people.

PubMed Central

Husebye, E; Skar, V; Høverstad, T; Melby, K

1992-01-01

Fifteen healthy old people mean age 84 years (range 80-91 years), were examined to assess the effect of advanced age on the microecology of the upper gastrointestinal tract. Twelve of 15 (80%) were hypochlorhydric with pH 6.6 (0.3) (mean (SEM) and a mean bacterial count of 10(8) colony forming units (CFU) per ml (range 10(5)-10(10)) in fasting gastric aspirate. Normochlorhydric subjects had low counts (< or = 10(1) CFU/ml). The microbial flora was dominated by viridans streptococci, coagulase negative staphylococci, and Haemophilus sp. Only one subject harboured significant concentrations of Gram negative bacilli with Escherichia coli (10(4-5) CFU/ml) and Klebsiella (10(4-5)). Strict anaerobes were not found. The total concentration of short chain fatty acids in gastric aspirate was 10.6 (2.9) mmol/l (mean (SEM). Absence of significant, intraluminal fermentation of xylose to CO2 was shown by the 14C-d Xylose breath test, and ambulatory manometry showed preserved fasting motility pattern of the small intestine. Serum immunoglobulins were normal. Advanced age is accompanied by fasting hypochlorhydria and colonisation with mainly Gram positive flora in the upper gut. Other factors than old age and fasting hypochlorhydria are required for colonisation with Gram negative bacilli. PMID:1446855

Phage inactivation of Staphylococcus aureus in fresh and hard-type cheeses.

PubMed

Bueno, Edita; García, Pilar; Martínez, Beatriz; Rodríguez, Ana

2012-08-01

Bacteriophages are regarded as natural antibacterial agents in food since they are able to specifically infect and lyse food-borne pathogenic bacteria without disturbing the indigenous microbiota. Two Staphylococcus aureus obligately lytic bacteriophages (vB_SauS-phi-IPLA35 and vB_SauS-phi-SauS-IPLA88), previously isolated from the dairy environment, were evaluated for their potential as biocontrol agents against this pathogenic microorganism in both fresh and hard-type cheeses. Pasteurized milk was contaminated with S. aureus Sa9 (about 10(6) CFU/mL) and a cocktail of the two lytic phages (about 10(6) PFU/mL) was also added. For control purposes, cheeses were manufactured without addition of phages. In both types of cheeses, the presence of phages resulted in a notorious decrease of S. aureus viable counts during curdling. In test fresh cheeses, a reduction of 3.83 log CFU/g of S. aureus occurred in 3h compared with control cheese, and viable counts were under the detection limits after 6h. The staphylococcal strain was undetected in both test and control cheeses at the end of the curdling process (24 h) and, of note, no re-growth occurred during cold storage. In hard cheeses, the presence of phages resulted in a continuous reduction of staphylococcal counts. In curd, viable counts of S. aureus were reduced by 4.64 log CFU/g compared with the control cheeses. At the end of ripening, 1.24 log CFU/g of the staphylococcal strain was still detected in test cheeses whereas 6.73log CFU/g was present in control cheeses. Starter strains were not affected by the presence of phages in the cheese making processes and cheeses maintained their expected physico-chemical properties. Copyright © 2012 Elsevier B.V. All rights reserved.

Ultra-low dose of Mycobacterium tuberculosis aerosol creates partial infection in mice.

PubMed

Saini, Divey; Hopkins, Gregory W; Seay, Sarah A; Chen, Ching-Ju; Perley, Casey C; Click, Eva M; Frothingham, Richard

2012-03-01

A murine low dose (LD) aerosol model is commonly used to test tuberculosis vaccines. Doses of 50-400 CFU (24h lung CFU) infect 100% of exposed mice. The LD model measures progression from infection to disease based on organ CFU at defined time points. To mimic natural exposure, we exposed mice to an ultra-low dose (ULD) aerosol. We estimated the presented dose by sampling the aerosol. Female C57BL/6 mice were exposed to Mycobacterium tuberculosis H37Rv aerosol at 1.0, 1.1, 1.6, 5.4, and 11 CFU presented dose, infecting 27%, 36%, 36%, 100%, and 95% of mice, respectively. These data are compatible with a stochastic infection event (Poisson distribution, weighted R(2)=0.97) or with a dose-response relationship (sigmoid distribution, weighted R(2)=0.97). Based on the later assumption, the ID50 was 1.6CFU presented dose (95% confidence interval, 1.2-2.1). We compared organ CFU after ULD and LD aerosols (5.4 vs. 395CFU presented dose). Lung burden was 30-fold lower in the ULD model at 4 weeks (3.4 vs. 4.8 logs, p<0.001) and 18 weeks (≤3.6 vs. 5.0 logs, p=0.01). Mice exposed to ULD aerosols as compared to LD aerosols had greater within-group CFU variability. Exposure to ULD aerosols leads to infection in a subset of mice, and to persistently low organ CFU. The ULD aerosol model may resemble human pulmonary tuberculosis more closely than the standard LD model, and may be used to identify host or bacterial factors that modulate the initial infection event. Copyright © 2011 Elsevier Ltd. All rights reserved.

The EmulSiv filter removes microbial contamination from propofol but is not a substitute for aseptic technique.

PubMed

Hall, Wendy C E; Jolly, Donald T; Hrazdil, Jiri; Galbraith, John C; Greacen, Maria; Clanachan, Alexander S

2003-01-01

To evaluate the ability of the EmulSiv filter (EF) to remove extrinsic microbial contaminants from propofol. Aliquots of Staphylococcus aureus (S. aureus), Candida albicans (C. albicans), Klebsiella pneumoniae (K. pneumoniae), Moraxella osloensis (M. osloensis), Enterobacter agglomerans (E. agglomerans), Escherichia coli (E. coli), Serratia marcescens (S. marcescens), Moraxella catarrhalis (M. catarrhalis), Haemophilus influenzae (H. influenzae) and Campylobacter jejuni (C. jejuni) were inoculated into vials containing 20 mL of sterile propofol. The unfiltered inoculated propofol solutions served as controls. Ten millilitres and 20 mL samples of the inoculated propofol were filtered through the EF. All solutions were then subplated onto three culture plates using a precision 1 micro L calibrated platinum loop and incubated. The number of colony forming units (CFU) were counted. Data were analyzed using a one-sample t test, and a P value of less than 0.05 was selected as the level of statistical significance. The EF was able to completely remove CFU of S. aureus, C. albicans, K. pneumoniae, M. osloensis, E. agglomerans, E. coli, S. marcescens, and M. catarrhalis (P < 0.05). A small number of H. influenzae CFU were able to evade filtration in both the 10 mL and 20 mL samples. C. jejuni CFU were able to evade filtration in only the 10 mL sample. The EF removes the majority of microbial contaminates from propofol with the exception of H. influenzae and C. jejuni. Although the EF is capable of removing most of the microbial contamination produced by H. influenzae and C. jejuni, a few CFU are capable of evading filtration. Consequently, even the use of a filter capable of removing microbial contaminants is not a substitute for meticulous aseptic technique and prompt administration when propofol is used.

Microbiological Quality and Safety of Fresh Fruits and Vegetables at Retail Levels in Korea.

PubMed

Tango, Charles Nkufi; Wei, Shuai; Khan, Imran; Hussain, Mohammad Shakhawat; Kounkeu, Paul-François Ngnitcho; Park, Joong-Hyun; Kim, Se-Hun; Oh, Deog Hwan

2018-02-01

The aim of this study was to evaluate the microbiological quality and safety of fresh produce at retail level in Korea in order to periodically update information and establish available risks associated with consumption of fresh fruits and vegetables. The samples from different markets located in 3 provinces of South Korea were collected. The protocol in the Korean Food Standards Codex was applied and generic Escherichia coli, coliforms, aerobic mesophilic bacteria (AMB), and yeast and mold (YM) in 360 packaged and unpackaged fresh fruits and vegetables were analyzed. Presence of pathogens was examined using real-time polymerase chain reaction (q-PCR) after enrichment of samples. For all, the microbial counts ranged from 1.7 to 10.6 log cfu/g for AMB, 2.2 to 7.9 log cfu/g for coliforms, and 5.5 to 7.9 log cfu/g for YM. Three lettuce samples were contaminated by E. coli with a bacterial load ranging from 2 to 4 log cfu/g. Salmonella spp. were not detected in any fresh produce. Listeria monocytogenes, E. coli O157:H7, and Staphylococcus aureus were found in 1 (0.6%), 3 (0.8%), and 5 (1.4%) fresh produce samples, respectively. Bacillus cereus (50.3%) and Clostridium perfringens (13.3%) had the highest prevalence. These results indicate the need for employing strict control measures and developing preventive strategies to improve the quality and safety of fresh produce in Korea. © 2018 Institute of Food Technologists®.

Inactivation of E. coli O157:H7 and nonpathogenic E. coli in strawberry juice by pulsed electric field, sodium benzoate, potassium sorbate, and citric acid

USDA-ARS?s Scientific Manuscript database

Introduction: Current regulations require that juice processors effect a 5 log CFU/ml reduction of a target pathogen prior to distributing products. Whereas thermal pasteurization reduces the sensory characteristics of juice by altering flavor components, pulsed electric field (PEF) treatment may ...

Appearance and overall acceptability of fresh-cut cantaloupe pieces from whole melon treated with wet steam process

USDA-ARS?s Scientific Manuscript database

Minimally processed fresh-cut fruits have a limited shelf-life because of deterioration caused by spoilage microflora and changes in physiological processes. Whole melons were inoculated with 7 log CFU/ml of each bacterium (Salmonella, Escherichia coli O157:H7 and Listeria monocytogenes) and then t...

Antimicrobials for reduction of Salmonella contamination in uncooked, surface-browned breaded chicken products.

PubMed

Moschonas, Galatios; Geornaras, Ifigenia; Stopforth, Jarret D; Wach, Damien; Woerner, Dale R; Belk, Keith E; Smith, Gary C; Sofos, John N

2012-06-01

Surface-browned but uncooked frozen breaded chicken products have been associated with salmonellosis outbreaks due to inadequate or no cooking of the products before consumption. This study was conducted to evaluate the effect of three antimicrobials against Salmonella during manufacture of a surface-browned, uncooked frozen breaded chicken meat product. Fresh chicken breast meat portions (5 by 5 by 5 cm) were inoculated (4 to 5 log CFU/g) with Salmonella and mixed with caprylic acid (CAA; 0.5 and 1.0%), carvacrol (CAR; 0.3 and 0.5%), ε-polylysine (POL; 0.125 and 0.25%), or distilled water (control). Sodium chloride (1.2%) and sodium tripolyphosphate (0.3%) were added to all treatments, and the mixtures were ground (5% total moisture enhancement level) and formed into portions (9 by 5 by 3 cm). The products were breaded and surface browned by baking in an oven (208°C for 15 min) or deep frying in vegetable oil (190°C for 15 s), packaged in polyethylene bags, and stored at -20°C for 7 days. Total reductions of inoculated Salmonella in untreated control oven- or fryer-browned products after frozen storage were 1.2 and 0.8 log CFU/g, respectively. In comparison, treatment with CAA, CAR, or POL reduced initial pathogen counts by 3.3 to >4.5, 4.1 to >4.7, and 1.1 to 1.6 log CFU/g, respectively, regardless of the antimicrobial concentration and browning method. Treatment with 1.0% CAA (oven browned) or 0.5% CAR (oven or fryer browned) reduced Salmonella to nondetectable levels (<0.3 log CFU/g) in stored frozen products. These data may be useful for development of suitable antimicrobial treatments to reduce the risk of Salmonella contamination in surface-browned, uncooked frozen breaded chicken products.

Reduction of Escherichia coli O157:H7 on produce by use of electrolyzed water under simulated food service operation conditions.

PubMed

Pangloli, Philipus; Hung, Yen-Con; Beuchat, Larry R; King, C Harold; Zhao, Zhi-Hui

2009-09-01

Treatment of fresh fruits and vegetables with electrolyzed water (EW) has been shown to kill or reduce foodborne pathogens. We evaluated the efficacy of EW in killing Escherichia coli O157:H7 on iceberg lettuce, cabbage, lemons, and tomatoes by using washing and/or chilling treatments simulating those followed in some food service kitchens. Greatest reduction levels on lettuce were achieved by sequentially washing with 14-A (amperage) acidic EW (AcEW) for 15 or 30 s followed by chilling in 16-A AcEW for 15 min. This procedure reduced the pathogen by 2.8 and 3.0 log CFU per leaf, respectively, whereas washing and chilling with tap water reduced the pathogen by 1.9 and 2.4 log CFU per leaf. Washing cabbage leaves for 15 or 30 s with tap water or 14-A AcEW reduced the pathogen by 2.0 and 3.0 log CFU per leaf and 2.5 to 3.0 log CFU per leaf, respectively. The pathogen was reduced by 4.7 log CFU per lemon by washing with 14-A AcEW and 4.1 and 4.5 log CFU per lemon by washing with tap water for 15 or 30 s. A reduction of 5.3 log CFU per lemon was achieved by washing with 14-A alkaline EW for 15 s prior to washing with 14-A AcEW for 15 s. Washing tomatoes with tap water or 14-A AcEW for 15 s reduced the pathogen by 6.4 and 7.9 log CFU per tomato, respectively. Application of AcEW using procedures mimicking food service operations should help minimize cross-contamination and reduce the risk of E. coli O157:H7 being present on produce at the time of consumption.

Lactobacillus plantarum BSGP201683 Isolated from Giant Panda Feces Attenuated Inflammation and Improved Gut Microflora in Mice Challenged with Enterotoxigenic Escherichia coli.

PubMed

Liu, Qian; Ni, Xueqin; Wang, Qiang; Peng, Zhirong; Niu, Lili; Wang, Hengsong; Zhou, Yi; Sun, Hao; Pan, Kangcheng; Jing, Bo; Zeng, Dong

2017-01-01

In this work, we searched for an effective probiotic that can help control intestinal infection, particularly enterotoxigenic Escherichia coli K88 (ETEC) invasion, in giant panda ( Ailuropoda melanoleuca ). As a potential probiotic strain, Lactobacillus plantarum BSGP201683 ( L. plantarum G83) was isolated from the feces of giant panda and proven beneficial in vitro . This study was aimed to evaluate the protective effect of L. plantarum G83 in mice challenged with ETEC. The mice were orally administered with 0.2 mL of PBS containing L. plantarum G83 at 0 colony-forming units (cfu) mL -1 (control; negative control, ETEC group), 5.0 × 10 8 cfu mL -1 (LDLP), 5.0 × 10 9 cfu mL -1 (MDLP), and 5.0 × 10 10 cfu mL -1 (HDLP) for 14 consecutive days. At day 15, the mice (LDLP, MDLP, HDLP, and ETEC groups) were challenged with ETEC and assessed at 0, 24, and 144 h. Animal health status; chemical and biological intestinal barriers; and body weight were measured. Results showed that L. plantarum G83 supplementation protected the mouse gut mainly by attenuating inflammation and improving the gut microflora. Most indices significantly changed at 24 h after challenge compared to those at 0 and 144 h. All treatment groups showed inhibited plasma diamine oxidase activity and D -lactate concentration. Tight-junction protein expression was down-regulated, and interleukin (IL)-1β, IL-6, IL-8, TLR4, and MyD88 levels were up-regulated in the jejunum in the LDLP and MDLP groups. The number of the Enterobacteriaceae family and the heat-labile enterotoxin (LT) gene decreased ( P < 0.05) in the colons in the LDLP and MDLP groups. All data indicated that L. plantarum G83 could attenuate acute intestinal inflammation caused by ETEC infection, and the low and intermediate doses were superior to the high dose. These findings suggested that L. plantarum G83 may serve as a protective probiotic for intestinal disease and merits further investigation.

Reduction of Bacterial Pathogens and Potential Surrogates on the Surface of Almonds Using High-Intensity 405-Nanometer Light.

PubMed

Lacombe, Alison; Niemira, Brendan A; Sites, Joseph; Boyd, Glenn; Gurtler, Joshua B; Tyrell, Breanna; Fleck, Melissa

2016-11-01

The disinfecting properties of high-intensity monochromatic blue light (MBL) were investigated against Escherichia coli O157:H7, Salmonella , and nonpathogenic bacteria inoculated onto the surface of almonds. MBL was generated from an array of narrow-band 405-nm light-emitting diodes. Almonds were inoculated with higher or lower levels (8 or 5 CFU/g) of pathogenic E. coli O157:H7 and Salmonella , as well as nonpathogenic E. coli K-12 and an avirulent strain of Salmonella Typhimurium, for evaluation as potential surrogates for their respective pathogens. Inoculated almonds were treated with MBL for 0, 1, 2, 4, 6, 8, and 10 min at a working distance of 7 cm. Simultaneous to treatment, cooling air was directed onto the almonds at a rate of 4 ft 3 /min (1.89 ×10 -3 m 3 /s), sourced through a container of dry ice. An infrared camera was used to monitor the temperature readings after each run. For E. coli K-12, reductions of up to 1.85 or 1.63 log CFU/g were seen for higher and lower inoculum levels, respectively; reductions up to 2.44 and 1.44 log CFU/g were seen for E. coli O157:H7 (higher and lower inoculation levels, respectively). Attenuated Salmonella was reduced by up to 0.54 and 0.97 log CFU/g, whereas pathogenic Salmonella was reduced by up to 0.70 and 0.55 log CFU/g (higher and lower inoculation levels, respectively). Inoculation level did not significantly impact minimum effective treatment times, which ranged from 1 to 4 min. Temperatures remained below ambient throughout treatment, indicating that MBL is a nonthermal antimicrobial process. The nonpathogenic strains of E. coli and Salmonella each responded to MBL in a comparable manner to their pathogenic counterparts. These results suggest that these nonpathogenic strains may be useful in experiments with MBL in which a surrogate is required, and that MBL warrants further investigation as a potential antimicrobial treatment for low-moisture foods.

Fate of Escherichia coli O157:H7 in manure compost-amended soil and on carrots and onions grown in an environmentally controlled growth chamber.

PubMed

Islam, Mahbub; Morgan, Jennie; Doyle, Michael P; Jiang, Xiuping

2004-03-01

Studies were done to determine the fate of Escherichia coli O157:H7 in manure compost-amended soil and on carrots and green onions grown in an environmentally controlled growth chamber. Commercial dairy cattle manure compost was inoculated with a five-strain mixture of green fluorescent protein-labeled E. coli O157:H7 at 10(7) CFU g(-1) and mixed with unsterilized Tifton sandy loam soil at a ratio of 1:5. Baby carrot or green onion seedlings were planted into the manure compost-amended soil in pots, and soil samples surrounding the plant, edible carrot roots and onion bulb samples, and soil immediately beneath the roots were assayed for E. coli O157:H7 in triplicate at weekly intervals for the first 4 weeks, and every 2 weeks for the remainder of the plant growth cycle (up to 3 months). E. coli O157:H7 cell numbers decreased within 64 days by 3 log CFU/g in soil and soil beneath the roots of green onions and by more than 2 log CFU/g on onions. E. coli O157:H7 survived better during the production of carrots, with a 2.3-log CFU/g reduction in soil and a 1.7-log CFU/g reduction on carrots within 84 days. These results indicate that the type of plant grown is an important factor influencing the survival of E. coli O157:H7 both on the vegetable and in the soil in which the vegetable is grown.

Quinolone Resistance Reversion by Targeting the SOS Response.

PubMed

Recacha, E; Machuca, J; Díaz de Alba, P; Ramos-Güelfo, M; Docobo-Pérez, F; Rodriguez-Beltrán, J; Blázquez, J; Pascual, A; Rodríguez-Martínez, J M

2017-10-10

Suppression of the SOS response has been postulated as a therapeutic strategy for potentiating antimicrobial agents. We aimed to evaluate the impact of its suppression on reversing resistance using a model of isogenic strains of Escherichia coli representing multiple levels of quinolone resistance. E. coli mutants exhibiting a spectrum of SOS activity were constructed from isogenic strains carrying quinolone resistance mechanisms with susceptible and resistant phenotypes. Changes in susceptibility were evaluated by static (MICs) and dynamic (killing curves or flow cytometry) methodologies. A peritoneal sepsis murine model was used to evaluate in vivo impact. Suppression of the SOS response was capable of resensitizing mutant strains with genes encoding three or four different resistance mechanisms (up to 15-fold reductions in MICs). Killing curve assays showed a clear disadvantage for survival (Δlog 10 CFU per milliliter [CFU/ml] of 8 log units after 24 h), and the in vivo efficacy of ciprofloxacin was significantly enhanced (Δlog 10 CFU/g of 1.76 log units) in resistant strains with a suppressed SOS response. This effect was evident even after short periods (60 min) of exposure. Suppression of the SOS response reverses antimicrobial resistance across a range of E. coli phenotypes from reduced susceptibility to highly resistant, playing a significant role in increasing the in vivo efficacy. IMPORTANCE The rapid rise of antibiotic resistance in bacterial pathogens is now considered a major global health crisis. New strategies are needed to block the development of resistance and to extend the life of antibiotics. The SOS response is a promising target for developing therapeutics to reduce the acquisition of antibiotic resistance and enhance the bactericidal activity of antimicrobial agents such as quinolones. Significant questions remain regarding its impact as a strategy for the reversion or resensitization of antibiotic-resistant bacteria. To address this question, we have generated E. coli mutants that exhibited a spectrum of SOS activity, ranging from a natural SOS response to a hypoinducible or constitutively suppressed response. We tested the effects of these mutations on quinolone resistance reversion under therapeutic concentrations in a set of isogenic strains carrying different combinations of chromosome- and plasmid-mediated quinolone resistance mechanisms with susceptible, low-level quinolone resistant, resistant, and highly resistant phenotypes. Our comprehensive analysis opens up a new strategy for reversing drug resistance by targeting the SOS response. Copyright © 2017 Recacha et al.

Efficacy of various antimicrobials on reduction of salmonella and campylobacter and quality attributes of ground chicken obtained from poultry parts treated in a postchill decontamination tank.

PubMed

Chen, Xi; Bauermeister, Laura J; Hill, Gretchen N; Singh, Manpreet; Bilgili, Sacit F; McKee, Shelly R

2014-11-01

Ground chicken is likely to have higher microbiological loads than whole carcasses and parts. Therefore, it is necessary to identify antimicrobials that reduce pathogens and overall microbial loads without negatively impacting meat quality. The objectives of this research were to evaluate the effect of various postchill antimicrobials on reducing Salmonella and Campylobacter, and determine the impact of these treatments on shelf life and quality attributes of ground chicken. Five treatments (0.003% chlorine, 0.07 and 0.1% peracetic acid [PAA], and 0.35 and 0.6% cetylpyridinium chloride [CPC]) were evaluated. Samples (n = 120) of skin-on chicken breast and thigh meat were inoculated with Salmonella Typhimurium (10(8) CFU/ml) and C. jejuni (10(8) CFU/ml). Following a 30-min attachment time, parts were rinsed with either chlorine, PAA, or CPC in a decontamination tank for 23 s. Parts then were ground, samples (25 g) were plated, and reduction of Salmonella Typhimurium and C. jejuni was determined. Noninoculated ground breast and thigh meat were used for sensory and shelf-life determination. Samples (n = 200) for shelf-life determination were collected on days 1, 4, 7, and 10 to estimate spoilage microflora of ground chicken stored at 4°C. Additionally, color measurement and sensory evaluation were conducted on days 1, 4, and 7. Ground chicken treated with 0.07 and 0.1% PAA had the greatest reductions (P ≤ 0.05) in Salmonella and Campylobacter providing approximately a 1.5-log reduction, followed by a 0.8-log reduction after treatment with 0.35 and 0.6% CPC. Chlorine (0.003%) was the least effective treatment (P ≤ 0.05), while treatments with 0.07 and 0.1% PAA also extended the shelf life of ground chicken for 3 days. None of the treatments had negative impact on color or sensory attributes of ground chicken patties during the storage (P ≤ 0.05). Results from this study indicated that using PAA as an antimicrobial agent in a postchill decontamination tank to treat ground poultry parts is effective for the reduction of Salmonella and Campylobacter while maintaining product quality.

High-level penicillin resistance and penicillin-gentamicin synergy in Enterococcus faecium.

PubMed Central

Torres, C; Tenorio, C; Lantero, M; Gastañares, M J; Baquero, F

1993-01-01

Thirty-seven Enterococcus faecium strains with different levels of penicillin susceptibility were studied in time-kill experiments with a fixed concentration (5 micrograms/ml) of gentamicin combined with different penicillin concentrations (6 to 600 micrograms/ml). Synergy was defined as a relative decrease in counts of greater than 2 log10 CFU per milliliter after 24 h of incubation when the combination of the antibiotics was compared with its most active component alone. The minimal synergistic penicillin concentrations found were 6 micrograms/ml for 16 of 16 strains for which penicillin MICs were < or = 25 micrograms/ml, 20 to 100 micrograms/ml for 14 of 17 strains for which penicillin MICs were 50 to 200 micrograms/ml, and 200 to 500 micrograms/ml for 4 of 4 strains for which MICs penicillin were > 200 micrograms/ml. Penicillin-gentamicin synergy was observed even in high-level penicillin-resistant E. faecium strains at penicillin concentrations close to one-half the penicillin MIC. The possibility of treating infections caused by high-level penicillin-resistant E. faecium strains with penicillin-gentamicin combinations in particular cases may depend on the penicillin levels attainable in vivo. PMID:8285628

Effect of Water Hardness on Efficacy of Sodium Hypochlorite Inactivation of Escherichia coli O157:H7 in Water.

PubMed

Swanson, Sara; Fu, Tong-Jen

2017-03-01

This study examined how the hardness of water affected the efficacy of sodium hypochlorite in inactivating Escherichia coli O157:H7 in water. Water was prepared at different degrees of total hardness (0, 50, 100, 200, 500, 1,000, 2,000, and 5,000 mg/liter CaCO 3 ). Inactivation was assessed at different levels of free chlorine (0, 0.2, 0.5, and 1.0 ppm) at 2 to 4°C and pH 6.5. Thirty milliliters of chlorinated water was inoculated with 6 log CFU/ml of E. coli O157:H7 and allowed to mix for 3, 10, 20, or 30 s. In the absence of sodium hypochlorite, no reduction in counts of E. coli O157:H7 was observed regardless of the degree of water hardness. However, in the presence of hard water, under certain chlorine concentrations and exposure times, the reduction of E. coli O157:H7 in chlorinated hard water was significantly less than the reduction observed in chlorinated deionized water. For example, after exposure to 0.5 ppm of free chlorine for 10 s, E. coli O157:H7 counts were reduced by 4.8 ± 1.4, 2.0 ± 1.3, 1.6 ± 0.7, 0.5 ± 0.7, and 0.0 ± 0.1 log CFU/ml in water containing 0, 100, 1,000, 2,000, and 5,000 mg/liter CaCO 3 , respectively. With the exception of 5,000 mg/liter CaCO 3 , the effect of water hardness was no longer visible after 20 s of exposure to 0.5 ppm of free chlorine. Also, hard water significantly lowered the efficacy of sodium hypochlorite at 3 s of exposure to 1.0 ppm of free chlorine. But after 20 s of exposure to 1.0 ppm of free chlorine, the impact of water hardness was no longer observed. This study demonstrated that water hardness can affect the germicidal efficacy of sodium hypochlorite, and such an impact may or may not be apparent depending on the condition of the solution and the treatment time at which the observation is made. Under the conditions typically seen in commercial produce washing operations, the impact of water hardness on chlorine efficacy is likely to be insignificant compared with that of organic load.

[Effects of antimicrobial peptide LL-37 expressed and purified from prokaryotes in the murine model of vaginal candidiasis].

PubMed

Wang, F; Huo, Y; Yin, L R; Sun, B; Zhang, P P

2016-07-25

To study the effects of antimicrobial peptide LL-37 expressed and purified from prokaryotes on candida albicans growth. (1)Thirty female Kunming mice were treated with estrogen and white candida yeast suspension were poured into vagina to establish a vulvovaginal candidiasis(VVC)murine model. After successful establishing the VVC mouse model, mice were randomly sorted into test group(n=15)and control group(n=15). Suspension(30 μl, 100 μg/ml)of recombinant peptide LL-37 expressed and purified in Prokaryotes was given by intravaginal administration to the test group for 5 days, while the same amount of phosphate buffered saline(PBS)was given to the control group.(2)Tweenty-four hours after treatment, the fungal burden and colony-forming unit(CFU)of vaginal fluids were evaluated. All mice were subsequently sacrificed and vaginal tissues were harvested for tissue homogenate preparation. ELISA was used to determine the levels of nterleukin-10(IL-10)and interferon-γ(IFN-γ)in the isolated vaginal tissues. (1)VVC mouse model was established successfully in this study. Vaginal mucosa congestion, edema, vaginal plica disappearing were obviously observed in the control group. After treatment with recombinant protein LL-37 vaginal mucosa has no obvious change in the test group.(2)Fungal burden and CFU of vaginal fluids were significantly lower in the test group [(4.8±1.0)×10(4) CFU/ml]than that in the control group [(8.5±2.1)×10(4) CFU/ml, P=0.017]. IFN-γ level of the test group was increased [(257±11)vs(197±4)pg/ml, P=0.000], while the level of IL-10 was reduced [(287 ± 15)vs(379 ± 17)pg/ml P=0.000] resulting in a the ratio of IFN-γ/IL-10 was in significantly higher in test group(0.892±0.008 vs 0.496±0.013, P=0.000). Recombinant protein LL-37 expressed and purified from prokaryotes inhibits the growth candida albicans and improves vaginal immunity by adjusting IFN-γ and IL-10 secretion in the VVC mouse model, highlighting the therapeutic potential of LL-37 for VVC.

Effect of smokeless tobacco products on human oral bacteria growth and viability

PubMed Central

Liu, Min; Jin, Jinshan; Pan, Hongmiao; Feng, Jinhui; Cerniglia, Carl E.; Yang, Maocheng; Chen, Huizhong

2017-01-01

To evaluate the toxicity of smokeless tobacco products (STPs) on oral bacteria, seven smokeless tobacco aqueous extracts (STAEs) from major brands of STPs and three tobacco-specific N-nitrosamines (TSNAs) were used in a growth and viability test against 38 oral bacterial species or subspecies. All seven STAEs showed concentration-dependent effects on the growth and viability of tested oral bacteria under anaerobic culture conditions, although there were strain-to-strain variations. In the presence of 1 mg/ml STAEs, the growth of 4 strains decreased over 0.32–2.14 log10 fold, while 14 strains demonstrated enhanced growth of 0.3–1.76 log10 fold, and the growth of 21 strains was not significantly affected. In the presence of 10 mg/ml STAEs, the growth of 17 strains was inhibited 0.3–2.11 log10 fold, 18 strains showed enhanced growth of 0.3–0.97 log10 fold, and 4 strains were not significantly affected. In the presence of 50 mg/ml STAEs, the growth of 32 strains was inhibited 0.3–2.96 log10 fold, 8 strains showed enhanced growth of 0.3–1.0 log10 fold, and 2 strains were not significantly affected. All seven STAEs could promote the growth of 4 bacterial strains, including Eubacterium nodatum, Peptostreptococcus micros, Streptococcus anginosus, and Streptococcus constellatus. Exposure to STAEs modulated the viability of some bacterial strains, with 21.1–66.5% decrease for 4 strains at 1 mg/ml, 20.3–85.7% decrease for 10 strains at 10 mg/ml, 20.0–93.3% decrease for 27 strains at 50 mg/ml, and no significant effect for 11 strains at up to 50 mg/ml. STAEs from snuffs inhibited more tested bacterial strains than those from snus indicating that the snuffs may be more toxic to the oral bacteria than snus. For TSNAs, cell growth and viability of 34 tested strains were not significantly affected at up to 100 μg/ml; while the growth of P. micros was enhanced 0.31–0.54 log10 fold; the growth of Veillonella parvula was repressed 0.33–0.36 log10 fold; and the cell viabilities of 2 strains decreased 56.6–69.9%. The results demonstrate that STAEs affected the growth of some types of oral bacteria, which may affect the healthy ecological balance of oral bacteria in humans. On the other hand, TSNAs did not significantly affect the growth of the oral bacteria. PMID:27756619

Alkaline phosphatase labeled SERS active sandwich immunoassay for detection of Escherichia coli

NASA Astrophysics Data System (ADS)

Bozkurt, Akif Goktug; Buyukgoz, Guluzar Gorkem; Soforoglu, Mehmet; Tamer, Ugur; Suludere, Zekiye; Boyaci, Ismail Hakki

2018-04-01

In this study, a sandwich immunoassay method utilizing enzymatic activity of alkaline phosphatase (ALP) on 5-bromo-4-chloro-3-indolyl phosphate (BCIP) for Escherichia coli (E. coli) detection was developed using surface enhanced Raman spectroscopy (SERS). For this purpose, spherical magnetic gold coated core-shell nanoparticles (MNPs-Au) and rod shape gold nanoparticles (Au-NRs) were synthesized and modified for immunomagnetic separation (IMS) of E. coli from the solution. In order to specify the developed method to ALP activity, Au-NRs were labeled with this enzyme. After successful construction of the immunoassay, BCIP substrate was added to produce the SERS-active product; 5-bromo-4-chloro-3-indole (BCI). A good linearity (R2 = 0.992) was established between the specific SERS intensity of BCI at 600 cm- 1 and logarithmic E. coli concentration in the range of 1.7 × 101-1.7 × 106 cfu mL- 1. LOD and LOQ values were also calculated and found to be 10 cfu mL- 1 and 30 cfu mL- 1, respectively.

Improving the AOAC use-dilution method by establishing a minimum log density value for test microbes on inoculated carriers.

PubMed

Tomasino, Stephen F; Pines, Rebecca M; Hamilton, Martin A

2009-01-01

The AOAC Use-Dilution methods, 955.14 (Salmonella enterica), 955.15 (Staphylococcus aureus), and 964.02 (Pseudomonas aeruginosa), are used to measure the efficacy of disinfectants on hard inanimate surfaces. The methods do not provide procedures to assess log density of the test microbe on inoculated penicylinders (carrier counts). Without a method to measure and monitor carrier counts, the associated efficacy data may not be reliable and repeatable. This report provides a standardized procedure to address this method deficiency. Based on carrier count data collected by four laboratories over an 8 year period, a minimum log density value is proposed to qualify the test results. Carrier count data were collected concurrently with 242 Use-Dilution tests. The tests were conducted on products bearing claims against P. aeruginosa and S. aureus with and without an organic soil load (OSL) added to the inoculum (as specified on the product label claim). Six carriers were assayed per test for a total of 1452 carriers. All 242 mean log densities were at least 6.0 (geometric mean of 1.0 x 10(6) CFU/carrier). The mean log densities did not exceed 7.5 (geometric mean of 3.2 x 10(7) CFU/carrier). For all microbes and OSL treatments, the mean log density (+/- SEM) was 6.7 (+/- 0.07) per carrier (a geometric mean of 5.39 x 10(6) CFU/carrier). The mean log density for six carriers per test showed good repeatability (0.29) and reproducibility (0.32). A minimum mean log density of 6.0 is proposed as a validity requirement for S. aureus and P. aeruginosa. The minimum level provides for the potential inherent variability that may be experienced by a wide range of laboratories and the slight effect due to the addition of an OSL. A follow-up report is planned to present data to support the carrier count procedure and carrier counts for S. enterica.

Effectiveness of disinfectant treatments for inactivating Piscirickettsia salmonis.

PubMed

Muniesa, A; Escobar-Dodero, J; Silva, N; Henríquez, P; Bustos, P; Perez, A M; Mardones, F O

2018-03-08

This short communication investigated in vitro differences between commercial disinfectants types (n = 36), doses of application, and time of action in the elimination of Piscirickettsia salmonis, the most important bacterium affecting farmed salmon in Chile. Seven different treatments were examined, including active and inactive chlorine dioxides, glutaraldehyde, hypochlorite disinfectants and detergents, peracetic acid, peroxides and other miscellaneous methods A 3 replicate set of each of the sample groups was stored at 20 °C and 95% relative humidity and retested after 1, 5 and 30 min with varying doses (low, recommended and high doses). Multiple comparison tests were performed for the mean log CFU/ml among different disinfectant types, dose (ppm) and time of exposure (minutes) on the reduction of P. salmonis. Overall, disinfection using peracetic acid, peroxides, and both active and inactive chlorine dioxides caused significantly higher reduction of >7.5 log CFU/ml in samples, compared to other tested sanitizers. The lowest reduction was obtained after disinfection with hypochlorite detergents. As expected, as doses and time of action increase, there was a significant reduction of the overall counts of P. salmonis. However, at lowest doses, only use of paracetic acids resulted in zero counts. Implementation of effective protocols, making use of adequate disinfectants, may enhance biosecurity, and ultimately, mitigate the impact of P. salmonis in farmed salmon. Copyright © 2018. Published by Elsevier B.V.

Effect of levofloxacin treatment on semen hyperviscosity in chronic bacterial prostatitis patients.

PubMed

Vicari, L O; Castiglione, R; Salemi, M; Vicari, B O; Mazzarino, M C; Vicari, E

2016-05-01

Changes in seminal fluid viscosity (SFV), reactive oxygen species (ROS) production, cytokines and seminal leucocyte concentration related to microbiological outcome in patients with chronic bacterial prostatitis (CBP) were studied. One hundred and ten infertile patients with CBP (positive sperm culture ≥10(5) colony-forming units [CFU] ml(-1), pathogens or Chlamydia in expressed prostatic secretions) were treated with levofloxacin 500 mg daily for 14 consecutive days per month for 3 months. In case of bacterial prostatitis, two conditions were examined: responders, eradication of 0 to <10(3) CFU ml(-1) (n = 78) and poor responders, >10(3) to <10(5) CFU ml(-1) (n = 32). Compared with poor responders, responders showed a significant increase of sperm progressive motility and a significant decrease in seminal leucocyte count, SFV, liquefaction time, ROS production (in all fractions and conditions), seminal tumour necrosis factor-α and interleukin 6. None of these variables showed significant differences compared with a control group of 37 fertile men. On the other hand, the poor responders showed significant changes in these variables compared with matched pretreatment values. In patients with CBP, antibiotic therapy alone leads to eradication in ≈71%, with improvement of sperm progressive motility, SFV and the framework of prooxidative factors. However, in the remaining ≈29% with poor antibiotic responsiveness, a deterioration of all variables is observed. © 2015 Blackwell Verlag GmbH.

Solar Disinfection of MODS Mycobacterial Cultures in Resource-Poor Settings

PubMed Central

Nathavitharana, Ruvandhi; Coronel, Jorge; Moore, David A. J.

2007-01-01

Introduction Safe disposal of TB culture material in which the infectious burden of clinical samples has been greatly amplified is an important challenge in resource-limited settings. The bactericidal capacity of solar cookers has been demonstrated previously for conventional bacteria and contaminated clinical waste. We investigated the use of a simple solar cooker for the sterilization of mycobacterial broth cultures from the microscopic observation drug susceptibility assay (MODS). Methods Simulated TB culture materials were prepared by inoculating 24-well MODS plates with 500 µL of a known concentration of Mycobacterium bovis BCG. In a series of experiments, samples were simultaneously placed inside a box-type solar cooker and control box and removed at timepoints between 15 minutes and 6 hours. Quantitative cultures were performed using retrieved samples to determine sterilization effect. Results All cultures from the control box were positive at or within 1–4 logs of inoculation concentration. Simulated culture plates at concentrations from 103colony-forming-units (CFU)/ml to 107 CFU/ml were completely sterilized after only one hour of cooker exposure, at temperatures between 50–102°C. At 109 CFU/ml (far in excess of diagnostic cultures), it was only possible to recover mycobacterial growth in plates removed after 15 minutes. By 30 minutes all plates were effectively sterilized. Discussion Solar disinfection provides a very effective, safe and low-cost alternative to conventional equipment used for disposal of mycobacterial culture material. Effect of climatic conditions and optimal operating procedure remain to be defined. PMID:17971863

Detection of shigella in lettuce by the use of a rapid molecular assay with increased sensitivity

PubMed Central

Jiménez, Kenia Barrantes; McCoy², Clyde B.; Achí, Rosario

2010-01-01

A Multiplex Polymerase Chain Reaction (PCR) assay to be used as an alternative to the conventional culture method in detecting Shigella and enteroinvasive Escherichia coli (EIEC) virulence genes ipaH and ial in lettuce was developed. Efficacy and rapidity of the molecular method were determined as compared to the conventional culture. Lettuce samples were inoculated with different Shigella flexneri concentrations (from 10 CFU/ml to 107 CFU/ml). DNA was extracted directly from lettuce after inoculation (direct-PCR) and after an enrichment step (enrichment PCR). Multiplex PCR detection limit was 104CFU/ml, diagnostic sensitivity and specificity were 100% accurate. An internal amplification control (IAC) of 100 bp was used in order to avoid false negative results. This method produced results in 1 to 2 days while the conventional culture method required 5 to 6 days. Also, the culture method detection limit was 106 CFU/ml, diagnostic sensitivity was 53% and diagnostic specificity was 100%. In this study a Multiplex PCR method for detection of virulence genes in Shigella and EIEC was shown to be effective in terms of diagnostic sensitivity, detection limit and amount of time as compared to Shigella conventional culture. PMID:24031579

Prevalence, quantification and antimicrobial resistance of Campylobacter spp. on chicken neck-skins at points of slaughter in 5 major cities located on 4 continents.

PubMed

Garin, Benoit; Gouali, Malika; Wouafo, Marguerite; Perchec, Anne-Marie; Pham, Minh Thu; Ravaonindrina, Noro; Urbès, Florence; Gay, Manu; Diawara, Abdoulaye; Leclercq, Alexandre; Rocourt, Jocelyne; Pouillot, Régis

2012-06-15

Quantitative data on Campylobacter contamination of food are lacking, notably in developing countries. We assessed Campylobacter contamination of chicken neck-skins at points of slaughter in 5 major cities in Africa (Dakar in Senegal, Yaounde in Cameroon), Oceania (Noumea in New Caledonia), the Indian Ocean (Antananarivo in Madagascar) and Asia (Ho Chi Minh City (HCMC) in Vietnam. One hundred and fifty slaughtered chickens were collected in each of the 5 major cities from semi-industrial abattoirs or markets (direct slaughter by the seller), and 65.5% (491/750) were found to be Campylobacter-positive. Two cities, Yaounde and Noumea, demonstrated high prevalence Campylobacter detection rates (92.7% and 96.7% respectively) in contrast with HCMC (15.3%). Four species were identified among 633 isolates, namely C. jejuni (48.3%), C. coli (37.3%), C. lari (11.7%) and C. upsaliensis (1%). HCMC was the only city with C. lari isolation as was Antananarivo for C. upsaliensis. C. coli was highly prevalent only in Yaounde (69.5%). Among the 491 samples positive in Campylobacter detection, 329 were also positive with the enumeration method. The number of Campylobacter colony-forming units (CFU) per gram of neck-skin in samples positive in enumeration was high (mean of the log(10): 3.2 log(10) CFU/g, arithmetic mean: 7900CFU/g). All the cities showed close enumeration means except HCMC with a 1.81 log(10) CFU/g mean for positive samples. Semi-industrial abattoir was linked to a significant lower count of Campylobacter contamination than direct slaughter by the seller (p=0.006). On 546 isolates (546/633, 86.3%) tested for antibiotic susceptibility, resistance to erythromycin, ampicillin and ciprofloxacin was observed for respectively 11%, 19% and 50%. HCMC was the city where antibiotic resistant rates were the highest (95%, p=0.014). Considering the 329 positive chickens in Campylobacter enumeration, the mean number of resistant isolates to at least 2 different antibiotic families (19.8%), may be estimated ca. 1500CFU/g; the corresponding mean of the log(10) would be 2.5 log(10)CFU/g. As chickens are sold at slaughter and brought directly at home to be cooked, these data suggest a high probability of cross-contamination. A substantial proportion of isolates are drug-resistant, which could lead to potential public health issues. Health authorities should consider measures to reduce Campylobacter contamination of chicken during farming and at slaughter, and to provide appropriate food hygiene education. Further studies are needed in particular to investigate food-handling practices in domestic kitchens. Copyright © 2012 Elsevier B.V. All rights reserved.

Effect of age of cook-in-bag delicatessen meats formulated with lactate-diacetate on the behavior of Listeria monocytogenes contamination introduced when opening the packages during storage.

PubMed

Geornaras, Ifigenia; Toczko, Darren; Sofos, John N

2013-07-01

This study evaluated the potential effect of age of cook-in-bag ham and turkey breast delicatessen meats formulated with lactate-diacetate on survival and/or growth of Listeria monocytogenes introduced after opening of packages and slicing of product. Commercially prepared cured ham and turkey breast products formulated with potassium lactate and sodium diacetate were stored at 1.7°C unsliced, in their original cook-in-bags, and without postlethality exposure. On days 5, 90, 120, and 180 of storage, product slices (10.2 by 7.6 cm) were surface inoculated (1 to 2 log CFU/cm²) with a 10-strain mixture of L. monocytogenes, vacuum packaged (seven slices per bag), and stored at 4°C for up to 13 weeks. Inoculated levels of L. monocytogenes on both products were 1.4 to 1.5 log CFU/cm². Irrespective of product age at slicing and inoculation, after 13 weeks of vacuum-packaged storage (4°C), pathogen counts on product slices were 1.5 to 2.3 (ham) and 2.3 to 2.5 (turkey) log CFU/cm². Overall, the results of the study showed that the age of the cook-in-bag products prior to slicing and inoculation with the pathogen did not (P ≥ 0.05) affect the behavior of L. monocytogenes during vacuum-packaged storage (4°C, up to 13 weeks) of ham and turkey slices. Mean counts of lactic acid bacteria and yeasts and molds, when detected, did not exceed approximately 1 and 2 log CFU/cm², respectively, among all stored samples. Findings of the study will be useful to the meat industry and risk assessors in their efforts to control L. monocytogenes in ready-to-eat meat products.

The human jejunum has an endogenous microbiota that differs from those in the oral cavity and colon.

PubMed

Sundin, Olof H; Mendoza-Ladd, Antonio; Zeng, Mingtao; Diaz-Arévalo, Diana; Morales, Elisa; Fagan, B Matthew; Ordoñez, Javier; Velez, Philip; Antony, Nishaal; McCallum, Richard W

2017-07-17

The upper half of the human small intestine, known as the jejunum, is the primary site for absorption of nutrient-derived carbohydrates, amino acids, small peptides, and vitamins. In contrast to the colon, which contains 10 11 -10 12 colony forming units of bacteria per ml (CFU/ml), the normal jejunum generally ranges from 10 3 to 10 5  CFU per ml. Because invasive procedures are required to access the jejunum, much less is known about its bacterial microbiota. Bacteria inhabiting the jejunal lumen have been investigated by classical culture techniques, but not by culture-independent metagenomics. The lumen of the upper jejunum was sampled during enteroscopy of 20 research subjects. Culture on aerobic and anaerobic media gave live bacterial counts ranging from 5.8 × 10 3 CFU/ml to 8.0 × 10 6 CFU/ml. DNA from the same samples was analyzed by 16S rRNA gene-specific quantitative PCR, yielding values from 1.5 × 10 5 to 3.1 × 10 7 bacterial genomes per ml. When calculated for each sample, estimated bacterial viability ranged from effectively 100% to a low of 0.3%. 16S rRNA metagenomic analysis of uncultured bacteria by Illumina MiSeq sequencing gave detailed microbial composition by phylum, genus and species. The genera Streptococcus, Prevotella, Veillonella and Fusobacterium, were especially abundant, as well as non-oral genera including Escherichia, Klebsiella, and Citrobacter. The jejunum was devoid of the genera Alistipes, Ruminococcus, Faecalibacterium, and other extreme anaerobes abundant in the colon. In patients with higher bacterial loads, there was no significant change in microbial species composition. The jejunal lumen contains a distinctive bacterial population consisting primarily of facultative anaerobes and oxygen-tolerant obligate anaerobes similar to those found in the oral cavity. However, the frequent abundance of Enterobacteriaceae represents a major difference from oral microbiota. Although a few genera are shared with the colon, we found no evidence for retrograde movement of the most abundant colonic microbes to the jejunum. Some individuals had much higher bacterial loads, but this was not correlated with decreases in bacterial species diversity or other evidence of dysbiosis.

Performance of point-of-care Xpert HIV-1 plasma viral load assay at a tertiary HIV care centre in Southern India.

PubMed

Swathirajan, Chinnambedu Ravichandran; Vignesh, Ramachandran; Boobalan, Jayaseelan; Solomon, Sunil Suhas; Saravanan, Shanmugam; Balakrishnan, Pachamuthu

2017-10-01

Sustainable suppression of HIV replication forms the basis of anti-retroviral therapy (ART) medication. Thus, reliable quantification of HIV viral load has become an essential factor to monitor the effectiveness of the ART. Longer turnaround-time (TAT), batch testing and technical skills are major drawbacks of standard real-time PCR assays. The performance of the point-of-care Xpert HIV-1 viral load assay was evaluated against the Abbott RealTime PCR m2000rt system. A total of 96 plasma specimens ranging from 2.5 log10 copies ml -1 to 4.99 log10 copies ml -1 and proficiency testing panel specimens were used. Precision and accuracy were checked using the Pearson correlation co-efficient test and Bland-Altman analysis. Compared to the Abbott RealTime PCR, the Xpert HIV-1 viral load assay showed a good correlation (Pearson r=0.81; P<0.0001) with a mean difference of 0.27 log10 copies ml -1 (95 % CI, -0.41 to 0.96 log10 copies ml -1 ; sd, 0.35 log10 copies ml -1 ). Reliable and ease of testing individual specimens could make the Xpert HIV-1 viral load assay an efficient alternative method for ART monitoring in clinical management of HIV disease in resource-limited settings. The rapid test results (less than 2 h) could help in making an immediate clinical decision, which further strengthens patient care.

Enhancing the antibacterial effect of 461 and 521 nm light emitting diodes on selected foodborne pathogens in trypticase soy broth by acidic and alkaline pH conditions.

PubMed

Ghate, Vinayak; Leong, Ai Ling; Kumar, Amit; Bang, Woo Suk; Zhou, Weibiao; Yuk, Hyun-Gyun

2015-06-01

Light emitting diodes (LEDs) with their antibacterial effect present a novel method for food preservation. This effect may be influenced by environmental conditions such as the pH of the food contaminated by the pathogen. Thus, it is necessary to investigate the influence of pH on the antibacterial effect of LEDs before their application to real food matrices. Escherichia coli O157:H7, Salmonella Typhimurium and Listeria monocytogenes in trypticase soy broth were illuminated using 10-W 461 (22.1 mW/cm(2)) and 521 nm (16 mW/cm(2)) LEDs at pH values of 4.5, 6.0, 7.3, 8.0 and 9.5 for 7.5 h at 15 °C. Using the 461 nm LEDs, the populations of E. coli O157:H7 decreased by 2.1 ± 0.02, 1.2 ± 0.08 and 4.1 ± 0.42 log CFU/ml at pH 4.5, 7.3 and 9.5 respectively, after a dosage of 596.7 J/cm(2). For L. monocytogenes, approximately a 5.8 ± 0.03 log reduction was observed after 238.7 J/cm(2) at pH 4.5 using the 461 nm LEDs, while the bacterial concentration was reduced by 1.8 ± 0.01 log at pH 9.5 after 596.7 J/cm(2). Bacterial inactivation using the 521 nm LEDs showed similar trends to the 461 nm LEDs at both acidic and alkaline pH conditions but with lower (1-2 log CFU/ml) reductions after 432 J/cm(2). Lower D-values were observed for L. monocytogenes when exposed to LEDs at acidic pH values, while the sensitivity of E. coli O157:H7 and S. Typhimurium to LED was markedly increased at an alkaline pH. Regardless of the pH at which the cultures were illuminated, the percentage of sublethal injury increased with the treatment time. These results highlight the enhanced antibacterial effect of the 461 nm LED under acidic and alkaline pH conditions, proving its potential to preserve foods as well as to have synergistic effect with acidic and alkaline antimicrobials. Copyright © 2014 Elsevier Ltd. All rights reserved.

Viability of and Escherichia coli O157:H7 and Listeria monocytogenes in a delicatessen appetizer (yogurt-based) salad as affected by citrus extract (Citrox©) and storage temperature.

PubMed

Tsiraki, Maria I; Yehia, Hany M; Elobeid, Tahra; Osaili, Tareq; Sakkas, Hercules; Savvaidis, Ioannis N

2018-02-01

The antimicrobial effect of citrus extract (at 1 mL/kg [C1] and 2 mL/kg [C2]) on naturally occurring microbiota and inoculated pathogens (E. coli O157:H7 and L. monocytogenes at ca. 6 log cfu/g) in the traditional Greek yogurt-based salad Tzatziki stored at 4, 10, or 21 °C, was examined. Lactic acid bacteria (LAB) were high (8.0-8.5 log cfu/g) and varied only minimally for both the control (untreated) and the citrus extract-treated salad samples, whereas the higher citrus extract concentration yielded the lowest yeast populations, irrespective of temperature, during the entire storage period. Populations of inoculated E. coli (6 log cfu/g) declined in both untreated and citrus extract-treated samples from day 0-70, 35, and 15 at 4, 10, and 21 °C, respectively. Citrus extract had a significant effect on the survival of the inoculated E. coli O157:H7, with reductions of 2.8-4.8 log cfu/g in the citrus extract-treated samples at the end of the storage period. Our data show that L. monocytogenes survived in both untreated and citrus extract-treated samples during the entire storage period, irrespective of the storage temperature. The higher concentration of citrus extract had a significant effect on the survival of L. monocytogenes in the treated samples, and reductions of 1.5-3.0 logs were noted on final day 70, 35 and 15 at 4, 10 and 21 °C, respectively. The results of our study demonstrated the potential of citrus extract as a natural compound that can control the growth of food-borne pathogenic bacteria, such as E. coli O157:H7 and L. monocytogenes in Tzatziki, a yogurt-based salad. Copyright © 2017 Elsevier Ltd. All rights reserved.

Efficacy of acidified sodium chlorite treatments in reducing Escherichia coli O157:H7 on Chinese cabbage.

PubMed

Inatsu, Yasuhiro; Bari, Md Latiful; Kawasaki, Susumu; Isshiki, Kenji; Kawamoto, Shinichi

2005-02-01

Efficacy of acidified sodium chlorite for reducing the population of Escherichia coli O157:H7 pathogens on Chinese cabbage leaves was evaluated. Washing leaves with distilled water could reduce the population of E. coli O157:H7 by approximately 1.0 log CFU/g, whereas treating with acidified chlorite solution could reduce the population by 3.0 log CFU/g without changing the leaf color. A similar level of reduction was achieved by washing with sodium chlorite solution containing various organic acids. However, acidified sodium chlorite in combination with a mild heat treatment reduced the population by approximately 4.0 log CFU/g without affecting the color, but it softened the leaves. Moreover, the efficacy of the washing treatment was similar at low (4 degrees C) and room (25 degrees C) temperatures, indicating that acidified sodium chloride solution could be useful as a sanitizer for surface washing of fresh produce.

Immobilized Hydrolytic Enzymes Exhibit Antibiofilm Activity Against Escherichia coli at Sub-Lethal Concentrations.

PubMed

Villa, Federica; Secundo, Francesco; Polo, Andrea; Cappitelli, Francesca

2015-07-01

The effects of two commercially available immobilized enzymes (namely the glycosidase pectinase and the protease subtilisin A) at sub-lethal concentrations were investigated in terms of their influence on biofilm genesis, on the composition of the biofilm matrix, and their antibiotic synergy against Escherichia coli biofilm, used as a model system of bacterial biofilms. The best antibiofilm performance of solid-supported hydrolases was obtained at the surface concentration of 0.022 and 0.095 U/cm(2) with a reduction of 1.2 and 2.3 log CFU/biofilm for pectinase and subtilisin, respectively. At these enzyme surface concentrations, the biocatalysts affected the structural composition of the biofilm matrix, impacting biofilm thickness. Finally, the immobilized hydrolases enhanced biofilm sensitivity to a clinically relevant concentration of the antibiotic ampicillin. At the final antibiotic concentration of 0.1 mg/ml, a reduction of 2 and 3.5 log10 units in presence of 0.022 Upectinase/cm(2) and 0.095 Usubtilisin/cm(2) was obtained, respectively, in comparison the antibiotic alone. Immobilized pectinase and subtilisin at sub-lethal concentrations demonstrated a great potential for antibiofilm applications.

Validation of cooking times and temperatures for thermal inactivation of Yersinia pestis strains KIM5 and CDC-A1122 in irradiated ground beef.

PubMed

Porto-Fett, Anna C S; Juneja, Vijay K; Tamplin, Mark L; Luchansky, John B

2009-03-01

Irradiated ground beef samples (ca. 3-g portions with ca. 25% fat) inoculated with Yersina pestis strain KIM5 (ca. 6.7 log CFU/g) were heated in a circulating water bath stabilized at 48.9, 50, 52.5, 55, 57.5, or 60 degrees C (120, 122, 126.5, 131, 135.5, and 140 degrees F, respectively). Average D-values were 192.17, 34.38, 17.11, 3.87, 1.32, and 0.56 min, respectively, with a corresponding z-value of 4.67 degrees C (8.41 degrees F). In related experiments, irradiated ground beef patties (ca. 95 g per patty with ca. 25% fat) were inoculated with Y. pestis strains KIMS or CDC-A1122 (ca. 6.0 log CFU/g) and cooked on an open-flame gas grill or on a clam-shell type electric grill to internal target temperatures of 48.9, 60, and 71.1 degrees C (120, 140, and 160 degrees F, respectively). For patties cooked on the gas grill, strain KIM5 populations decreased from ca. 6.24 to 4.32, 3.51, and < or = 0.7 log CFU/g at 48.9, 60, and 71.1 degrees C, respectively, and strain CDC-A1122 populations decreased to 3.46 log CFU/g at 48.9 degrees C and to < or = 0.7 log CFU/g at both 60 and 71.1 degrees C. For patties cooked on the clam-shell grill, strain KIM5 populations decreased from ca. 5.96 to 2.53 log CFU/g at 48.9 degrees C and to < or = 0.7 log CFU/g at 60 or 71.1 degrees C, and strain CDC-A1122 populations decreased from ca. 5.98 to < or = 0.7 log CFU/g at all three cooking temperatures. These data confirm that cooking ground beef on an open-flame gas grill or on a clam-shell type electric grill to the temperatures and times recommended by the U.S. Department of Agriculture and the U.S. Food and Drug Administration Food Code, appreciably lessens the likelihood, severity, and/or magnitude of consumer illness if the ground beef were purposefully contaminated even with relatively high levels of Y. pestis.

Quantitation of bacteria through adsorption of intracellular biomolecules on carbon paste and screen-printed carbon electrodes and voltammetry of redox-active probes.

PubMed

Obuchowska, Agnes

2008-03-01

A new electrochemical method for the quantitation of bacteria that is rapid, inexpensive, and amenable to miniaturization is reported. Cyclic voltammetry was used to quantitate M. luteus, C. sporogenes, and E. coli JM105 in exponential and stationary phases, following exposure of screen-printed carbon working electrodes (SPCEs) to lysed culture samples. Ferricyanide was used as a probe. The detection limits (3s) were calculated and the dynamic ranges for E. coli (exponential and stationary phases), M. luteus (exponential and stationary phases), and C. sporogenes (exponential phase) lysed by lysozyme were 3 x 10(4) to 5 x 10(6) colony-forming units (CFU) mL(-1), 5 x 10(6) to 2 x 10(8) CFU mL(-1) and 3 x 10(3) to 3 x 10(5) CFU mL(-1), respectively. Good overlap was obtained between the calibration curves when the electrochemical signal was plotted against the dry bacterial weight, or between the protein concentration in the bacterial lysate. In contrast, unlysed bacteria did not change the electrochemical signal of ferricyanide. The results indicate that the reduction of the electrochemical signal in the presence of the lysate is mainly due to the fouling of the electrode by proteins. Similar results were obtained with carbon-paste electrodes although detection limits were better with SPCEs. The method described herein was applied to quantitation of bacteria in a cooling tower water sample.

Monte Carlo simulation of the risk of contamination of apples with Escherichia coli O157:H7.

PubMed

Duffy, Siobain; Schaffner, Donald W

2002-10-25

Quantitative descriptions of the frequency and extent of contamination of apple cider with pathogenic bacteria were obtained using literature data and computer simulation. Probability distributions were chosen to describe the risk of apple contamination by each suspected pathway. Tree-picked apples may be contaminated by birds infected with Escherichia coli O157:H7 when orchards were located near a sewage source (ocean or landfill). Dropped apples could become contaminated from either infected animal droppings or from contaminated manure if used as fertilizer. A risk assessment model was created in Analytica. The results of worst-case simulations revealed that 6-9 log CFU E. coli O157:H7 might be found on a harvest of 1000 dropped apples, while 3-4 log CFU contamination could be present on 1000 tree-picked apples. This model confirms that practices such as using dropped apples and using animal waste as fertilizer increase risk in the production of apple cider, and that pasteurization may not eliminate all contamination in juice from heavily contaminated fruit. Recently published FDA regulations for juices requiring a 5-log CFU/ml reduction of pathogenic bacteria in fresh juices should be a fail-safe measure for apples harvested in all but the worst-case scenarios.

Use of phytic acid and hyper-salting to eliminate Escherichia coli O157:H7 from napa cabbage for kimchi production in a commercial plant.

PubMed

Kim, Nam Hee; Jang, Seong Ho; Kim, Soon Han; Lee, Hee Jung; Kim, Younghoon; Ryu, Jee Hoon; Rhee, Min Suk

2015-12-02

The aim of this study was to develop a new salting method using natural phytic acid (PA) to ensure the microbiological safety and quality of salted napa cabbage used for kimchi production. The production of salted napa cabbage involves several stages: trimming, hyper-salting (20% NaCl) for up to 1h, salting (10% NaCl for 10-18 h), three sequential washes in water (30s for each), and draining (2 h). Two separate experiments were performed: one to determine the appropriate treatment conditions and a second to validate applicability under commercial conditions. In Experiment I, the effects of hyper-salting with PA on Escherichia coli O157:H7 numbers were tested in the laboratory. The following variables were monitored: 1) PA concentration (1, 2, 3%, w/w); 2) the ratio of the sample weight to the total volume of the solution (1:1.5, 1:3, or 1:6); 3) the hyper-salting time (30 or 60 min); and 4) the salting time (2, 5, or 8 h). A procedure that achieved a >5-log reduction in the E. coli O157:H7 population was then tested in an actual kimchi processing plant (Experiment II). The results from Experiment I showed that bactericidal efficacy increased as all the measured variables increased (p<0.05). Hyper-salting with 2% PA at a sample-to-water ratio (w/v) of 1:3 and 1:6 for 60 min resulted in a >5-log CFU/g reduction in the E. coli O157:H7 population. Further salting for 5h completely eliminated (<1-log CFU/g) all bacteria. Thus, hyper-salting with PA 2% at a sample-to-water ratio of 1:3 for 60 min, followed by salting for 5h, was tested under large-scale production conditions. The results revealed that the initial aerobic plate counts (APC), total coliform counts (TC), and fecal coliform counts (FC) were 6.6, 3.4, and 2.8-log CFU/g, respectively. The selected protocol reduced these values by 3.7-, >2.4-, and >1.8-log CFU/g, respectively. The 5h salting step maintained the TC and FC at <1-log CFU/g; however, the APC recovered somewhat. The pH and salinity of the treated salted napa cabbages were within the ranges required for kimchi production (pH 5.1-5.3 and 1.5-2.0%, respectively). These results suggest that this novel method of salting food ensures microbiological safety and reduces the production time. Copyright © 2015 Elsevier B.V. All rights reserved.

Water quality status of dugouts from five districts in Northern Ghana: implications for sustainable water resources management in a water stressed tropical savannah environment.

PubMed

Cobbina, Samuel J; Anyidoho, Louis Y; Nyame, Frank; Hodgson, I O A

2010-08-01

This study was primarily aimed at investigating the physicochemical and microbial quality of water in 14 such dugouts from five districts in the northern region of Ghana. Results obtained suggest that except for colour, turbidity, total iron and manganese, many physicochemical parameters were either within or close to the World Health Organisation's acceptable limits for drinking water. Generally, colour ranged from 5 to 750 Hz (mean 175 Hz), turbidity from 0.65 to 568 nephelometric turbidity units (NTU; mean 87.9 NTU), total iron from 0.07 to 7.85 mg/L (mean 1.0 mg/L) and manganese from 0.03 to 1.59 mg/L (mean 0.50 mg/L). Coliform counts in water from all the dugouts in both wet and dry seasons were, however, above the recommended limits for drinking water. Total and faecal coliforms ranged from 125 to 68,000 colony forming units (cfu)/100 mL (mean 10,623 cfu/100 mL) and <1 to 19,000 cfu/100 mL (mean 1,310 cfu /100 mL), respectively. The poor microbial quality, as indicated by the analytically significant presence of coliform bacteria in all samples of dugout water, strongly suggests susceptibility and exposure to waterborne diseases of, and consequent health implications on, the many people who continuously patronise these vital water resources throughout the year. In particular, more proactive sustainable water management options, such as introduction to communities of simple but cost-effective purification techniques for water drawn from dugouts for drinking purposes, education and information dissemination to the water users to ensure environmentally hygienic practices around dugouts, may be needed.

Biofilms of Enterococcus faecalis and Enterococcus faecium isolated from the processing of ricotta and the control of these pathogens through cleaning and sanitization procedures.

PubMed

da Silva Fernandes, Meg; Kabuki, Dirce Yorika; Kuaye, Arnaldo Yoshiteru

2015-05-04

The biofilm formation of Enterococcus faecalis and Enterococcus faecium isolated from the processing of ricotta on stainless steel coupons was evaluated, and the effect of cleaning and sanitization procedures in the control of these biofilms was determined. The formation of biofilms was observed while varying the incubation temperature (7, 25 and 39°C) and time (0, 1, 2, 4, 6 and 8 days). At 7°C, the counts of E. faecalis and E. faecium were below 2 log10 CFU/cm(2). For the temperatures of 25 and 39°C, after 1 day, the counts of E. faecalis and E. faecium were 5.75 and 6.07 log10 CFU/cm(2), respectively, which is characteristic of biofilm formation. The tested sanitation procedures a) acid-anionic tensioactive cleaning, b) anionic tensioactive cleaning+sanitizer and c) acid-anionic tensioactive cleaning+sanitizer were effective in removing the biofilms, reducing the counts to levels below 0.4 log10 CFU/cm(2). The sanitizer biguanide was the least effective, and peracetic acid was the most effective. These studies revealed the ability of enterococci to form biofilms and the importance of the cleaning step and the type of sanitizer used in sanitation processes for the effective removal of biofilms. Copyright © 2015 Elsevier B.V. All rights reserved.

Efficacy of gaseous chlorine dioxide as a sanitizer for killing Salmonella, yeasts, and molds on blueberries, strawberries, and raspberries.

PubMed

Sy, Kaye V; McWatters, Kay H; Beuchat, Larry R

2005-06-01

Gaseous chlorine dioxide (ClO2) was tested for its effectiveness in killing Salmonella, yeasts, and molds on blueberries, strawberries, and red raspberries. An inoculum (100 microl, 6.0 to 6.8 log CFU/g of fruit) that contained five serotypes of Salmonella enterica was deposited on the skin, calyx tissue, or stem scar tissue of blueberries, skin or stem scar tissue of strawberries, and skin of red raspberries, dried for 2 h at 22 degrees C, then held for 20 h at 4 degrees C and 2 h at 22 degrees C before treatment. Sachets that contained reactant chemicals were formulated to release gaseous ClO2 at concentrations of 4.1, 6.2, and 8.0 mg/ liter of air within treatment times of 30, 60, and 120 min, respectively, at 23 +/- 1 degrees C. Lethality of ClO2 to Salmonella, yeasts, and molds was measured when fruits were in an atmosphere that contained 75 to 90% relative humidity. Treatment with 8.0 mg/liter of ClO2 significantly (alpha = 0.05) reduced the population of Salmonella on blueberries by 2.4 to 3.7 log CFU/g. Lethality was higher to cells in inoculum placed on the skin compared with the stem scar tissue. Populations of Salmonella on strawberries treated with 8.0 mg/liter of ClO2 were reduced by 3.8 to 4.4 log CFU/g; a significant reduction of 1.5 log CFU/g of raspberries was achieved. Treatment with 4.1 to 8.0 mg/liter of ClO2 caused reductions in populations of yeast and molds on blueberries, strawberries, and raspberries of 1.4 to 2.5, 1.4 to 4.2, and 2.6 to 3.0 log CFU/g, respectively. Treatment with 4.1 mg/liter of ClO2 did not markedly affect the sensory quality of fruits stored for up to 10 days at 8 degrees C. Results indicate that gaseous ClO2 has promise as a sanitizer for small fruits.

Antimicrobial activity of allyl isothiocyanate used to coat biodegradable composite films as affected by storage and handling conditions.

PubMed

Li, Weili; Liu, Linshu; Jin, Tony Z

2012-12-01

We evaluated the effects of storage and handling conditions on the antimicrobial activity of biodegradable composite films (polylactic acid and sugar beet pulp) coated with allyl isothiocyanate (AIT). Polylactic acid and chitosan were incorporated with AIT and used to coat one side of the film. The films were subjected to different storage conditions (storage time, storage temperature, and packed or unpacked) and handling conditions (washing, abrasion, and air blowing), and the antimicrobial activity of the films against Salmonella Stanley in tryptic soy broth was determined. The films (8.16 μl of AIT per cm(2) of surface area) significantly (P < 0.05) inhibited the growth of Salmonella during 24 h of incubation at 22°C, while the populations of Salmonella in controls increased from ca. 4 to over 8 log CFU/ml, indicating a minimum inactivation of 4 log CFU/ml on films in comparison to the growth on controls. Statistical analyses indicated that storage time, storage temperature, and surface abrasion affected the antimicrobial activity of the films significantly (P < 0.05). However, the differences in microbial reduction between those conditions were less than 0.5 log cycle. The results suggest that the films' antimicrobial properties are stable under practical storage and handling conditions and that these antimicrobial films have potential applications in food packaging.

Effects of essential oils of oregano and nutmeg on growth and survival of Yersinia enterocolitica and Listeria monocytogenes in barbecued chicken.

PubMed

Firouzi, R; Shekarforoush, S S; Nazer, A H K; Borumand, Z; Jooyandeh, A R

2007-11-01

The in vitro effects of plant essential oils (EOs) against pathogenic bacteria are well known, yet few studies have addressed the effects of these compounds against pathogens associated with ready-to-cook foods. Experiments were conducted to determine the effectiveness of oregano and nutmeg EOs on the growth and survival of Yersinia enterocolitica and Listeria monocytogenes in broth culture and in Iranian barbecued chicken. Ready-to-cook Iranian barbecued chicken was prepared according to the common practice with 1, 2, and 3 microl/g of oregano and nutmeg EOs. The test and control (without EOs) samples were inoculated with Y. enterocolitica and L. monocytogenes to a final concentration of 6 to 7 log CFU/g and stored at 3, 8, and 20 degrees C. Microorganisms were counted just before and at 24, 48, and 72 h after storage based on growth on Yersinia selective agar supplemented with cefsulodine, igrasan, and novobiocin and on Listeria selective agar supplemented with nalidixic acid and acriflavin. In the broth culture system, the nutmeg EO had a greater effect on L. monocytogenes (MIC = 0.20 nicrol/ml) than did the oregano EO (MIC = 0.26 microl/ml). However, the oregano EO had a greater effect on Y. enterocolitica (MIC = 0.16 microl/ml) than did the nutmeg EO (MIC = 0.25 microl/ml). In ready-to-cook Iranian barbecued chicken, the log CFU per gram of both bacteria after up to 72 h of incubation was not decreased significantly by various combinations of oregano and nutmeg EOs (1, 2, and 3 microl/g) and storage temperatures (3, 8, and 20 degrees C) when compared with control samples (without EOs). Although examination of spices in culture media can yield accurate microbiological data, without complementary tests in foods these data are of limited value for assessing food safety.

Application of edible coating with starch and carvacrol in minimally processed pumpkin.

PubMed

Santos, Adriele R; da Silva, Alex F; Amaral, Viviane C S; Ribeiro, Alessandra B; de Abreu Filho, Benicio A; Mikcha, Jane M G

2016-04-01

The present study evaluated the effect of an edible coating of cassava starch and carvacrol in minimally processed pumpkin (MPP). The minimal inhibitory concentration (MIC) of carvacrol against Escherichia coli, Salmonella enterica serotype Typhimurium, Aeromonas hydrophila, and Staphylococcus aureus was determined. The edible coating that contained carvacrol at the MIC and 2 × MIC was applied to MPP, and effects were evaluated with regard to the survival of experimentally inoculated bacteria and autochthonous microflora in MPP. Total titratable acidity, pH, weight loss, and soluble solids over 7 days of storage under refrigeration was also analyzed. MIC of carvacrol was 312 μg/ml. Carvacrol at the MIC reduced the counts of E. coli and S. Typhimurium by approximately 5 log CFU/g. A. hydrophila was reduced by approximately 8 log CFU/g, and S. aureus was reduced by approximately 2 log CFU/g on the seventh day of storage. Carvacrol at the 2 × MIC completely inhibited all isolates on the first day of Storage. coliforms at 35 °C and 45 °C were not detected (< 3 MPN/g) with either treatment on all days of shelf life. The treatment groups exhibited a reduction of approximately 2 log CFU/g in psychrotrophic counts compared with controls on the last day of storage. Yeast and mold were not detected with either treatment over the same period. The addition of carvacrol did not affect total titratable acidity, pH, or soluble solids and improved weight loss. The edible coating of cassava starch with carvacrol may be an interesting approach to improve the safety and microbiological quality of MPP.

Occurrence and characterization of Campylobacter in the Brazilian production and processing of broilers.

PubMed

Kuana, S L; Santos, L R; Rodrigues, L B; Borsoi, A; Moraes, H L S; Salle, C T P; Nascimento, V P

2008-12-01

Twenty-two commercial broiler flocks and their carcasses, totaling 546 samples (450 collected from a poultry farm and 96 from a slaughterhouse), were surveyed for the presence of Campylobacter. The positive results for Campylobacter among the analyzed samples were homogeneous, yielding 81.8% for cecal droppings, 80.9% for feces, and 80.4% for cloacal swabs. Pre-enrichment and direct plating showed that 77.85% and 81.8% of cloacal swabs, respectively, were positive for Campylobacter compared to 99.0% and 97.9% of carcasses testing positive with the pre-enrichment and direct plating methods. The Campylobacter count averaged 7.0 log10 colony-forming units (CFU)/g in cecal droppings, 5.15 log10 CFU/carcass after defeathering, and 4.24 log10 CFU/carcass after chilling. The samples were identified by the API Campy system as Campylobacter jejuni subsp. jejuni (68.8%), Campylobacter coli (8.3%), Campylobacter jejuni subsp. doylei (6.3%), Campylobacter upsaliensis (4.2%), and Campylobacter fetus subsp. fetus (2.1%). The analyzed broiler flocks were positive for Campylobacter in 81.8% of the cases, thus characterizing the occurrence of this pathogen in a broiler-producing region in southern Brazil. These results highlight the importance of programs targeted at the reduction of Campylobacter in poultry products, in order to minimize the risks for consumers.

Pulsed-plasma gas-discharge inactivation of microbial pathogens in chilled poultry wash water.

PubMed

Rowan, N J; Espie, S; Harrower, J; Anderson, J G; Marsili, L; MacGregor, S J

2007-12-01

A pulsed-plasma gas-discharge (PPGD) system was developed for the novel decontamination of chilled poultry wash water. Treatment of poultry wash water in the plasma generation chamber for up to 24 s at 4 degrees C reduced Escherichia coli NCTC 9001, Campylobacter jejuni ATCC 33560, Campylobacter coli ATCC 33559, Listeria monocytogenes NCTC 9863, Salmonella enterica serovar Enteritidis ATCC 4931, and S. enterica serovar Typhimurium ATCC 14028 populations to non-detectable levels (< or = 8 log CFU/ml). Although similar PPGD treatments at 4 degrees C also produced significant reductions (> or = 3 log CFU/ml) in recalcitrant B. cereus NCTC 11145 endospore numbers within 30 s, the level of endospore reduction was dependent on the nature of the sparged gas used in the plasma treatments. Scanning electron microscopy revealed that significant damage occurred at the cellular level in PPGD-treated test organisms. This electrotechnology delivers energy in intense ultrashort bursts, generating products such as ozone, UV light, acoustic and shock waves, and pulsed electric fields that have multiple bactericidal properties. This technology offers an exciting complementary or alternative approach for treating raw poultry wash water and for preventing cross-contamination in processing environments.

Enumeration of verocytotoxigenic Escherichia coli (VTEC) O157 and O26 in milk by quantitative PCR.

PubMed

Mancusi, Rocco; Trevisani, Marcello

2014-08-01

Quantitative real-time polymerase chain reaction (qPCR) can be a convenient alternative to the Most Probable Number (MPN) methods to count VTEC in milk. The number of VTEC is normally very low in milk; therefore with the aim of increasing the method sensitivity a qPCR protocol that relies on preliminary enrichment was developed. The growth pattern of six VTEC strains (serogroups O157 and O26) was studied using enrichment in Buffered Peptone Water (BPW) with or without acriflavine for 4-24h. Milk samples were inoculated with these strains over a five Log concentration range between 0.24-0.50 and 4.24-4.50 Log CFU/ml. DNA was extracted from the enriched samples in duplicate and each extract was analysed in duplicate by qPCR using pairs of primers specific for the serogroups O157 and O26. When samples were pre-enriched in BPW at 37°C for 8h, the relationship between threshold cycles (CT values) and VTEC Log numbers was linear over a five Log concentration range. The regression of PCR threshold cycle numbers on VTEC Log CFU/ml had a slope coefficient equal to -3.10 (R(2)=0.96) which is indicative of a 10-fold difference of the gene copy numbers between samples (with a 100 ± 10% PCR efficiency). The same 10-fold proportion used for inoculating the milk samples with VTEC was observed, therefore, also in the enriched samples at 8h. A comparison of the CT values of milk samples and controls revealed that the strains inoculated in milk grew with 3 Log increments in the 8h enrichment period. Regression lines that fitted the qPCR and MPN data revealed that the error of the qPCR estimates is lower than the error of the estimated MPN (r=0.982, R(2)=0.965 vs. r=0.967, R(2)=0.935). The growth rates of VTEC strains isolated from milk should be comparatively assessed before qPCR estimates based on the regression model are considered valid. Comparative assessment of the growth rates can be done using spectrophotometric measurements of standardized cultures of isolates and reference strains cultured in BPW at 37°C for 8h. The method developed for the serogroups O157 and O26 can be easily adapted to the other VTEC serogroups that are relevant for human health. The qPCR method is less laborious and faster than the standard MPN method and has been shown to be a good technique for quantifying VTEC in milk. Copyright © 2014 Elsevier B.V. All rights reserved.

Dynamic of Campylobacter Species Contamination Along a Poultry Slaughtering Chain

PubMed Central

Dib, Hussein; Mrad, Rachelle; Chami, Christelle; Jalkh, Rita

2014-01-01

The prevalence of Campylobacters was studied in a poultry farm and along the slaughtering chain. Fifteen swabs from a farm and 75 samples (swabs and rinsates) from its slaughterhouse were collected. All the faecal and cloacal farm swabs were contaminated by Campylobacter jejuni and C. coli against 50% for breast swabs. C. jejuni had a concentration of 6.26, 6.34 and 5.38 Log10 CFU/mL in faecal, cloacal and breast swabs respectively. Rinsates showed an almost constant concentration of Campylobacters (6 Log10 CFU/mL) with a predominance of the presumptive C. jejuni. C. lari was found in 22% of eviscerated samples. Faecal coliforms and E. coli, used as indicators, were detected in all samples (5.46 and 5.15 Log10 CFU/mL, respectively). Final chilling and chlorine (50 ppm) treatments decreased them to acceptable levels, unlike for Campylobacters. Further investigation of the dynamics of Campylobacters and their response to prevention and treatment measures is required. PMID:27800361

Decontamination of green onions and baby spinach by vaporized ethyl pyruvate.

PubMed

Durak, M Zeki; Churey, John J; Gates, Matthew; Sacks, Gavin L; Worobo, Randy W

2012-06-01

Foodborne illnesses associated with fresh produce continue to be a major concern as consumer demand for healthier and nonthermally processed food increases. The objective of this study was to evaluate vaporized ethyl pyruvate (EP; CAS 617-35-6) as a safe alternative antimicrobial agent for the decontamination of Escherichia coli O157:H7 on green onions and spinach. Baby spinach leaves and green onions were inoculated with a five-strain cocktail of E. coli O157:H7 (pGFP) by the dipping method. Samples were treated with concentrations of 0, 42, 105, and 420 mg/liter vaporized EP in a 2.6-liter enclosed container. The efficacy of EP vapors for reducing E. coli O157:H7((GFP)) populations on green onions and baby spinach at 4 and 10°C was monitored for 7 and 5 days, respectively. The lowest EP concentration (42 mg/liter) resulted in a 1.7-log reduction of E. coli O157:H7((GFP)) on green onions after 7 days at 4°C and a 1.9-log reduction after 5 days at 10°C (P < 0.05). In baby spinach, the same concentration resulted in 0.9-log and 1.4-log reductions (P < 0.05) of E. coli O157:H7((GFP)) after 7 days at 4°C and 5 days at 10°C, respectively. On green onions, the highest concentration of EP (420 mg/liter) reduced the population of E. coli O157:H7((GFP)) by >4.7 log CFU/g after 7 days at 4°C and 5 days at 10°C. The same concentration was also effective for reducing E. coli O157:H7((GFP)) populations in baby spinach by 4.3 log CFU/g after 7 days at 4°C and by >6.5 log CFU/g after 3 days at 10°C. Although the successful EP treatments minimally affected the sensory attributes of green onions, the treatments resulted in significant changes in the sensory attributes of baby spinach samples stored at 4 and 10°C. These results indicate that EP is an effective antimicrobial that could be used to enhance the safety of fresh produce depending on the sensory characteristics of the product.

Combined steam-ultrasound treatment of 2 seconds achieves significant high aerobic count and Enterobacteriaceae reduction on naturally contaminated food boxes, crates, conveyor belts, and meat knives.

PubMed

Musavian, Hanieh S; Butt, Tariq M; Larsen, Annette Baltzer; Krebs, Niels

2015-02-01

Food contact surfaces require rigorous sanitation procedures for decontamination, although these methods very often fail to efficiently clean and disinfect surfaces that are visibly contaminated with food residues and possible biofilms. In this study, the results of a short treatment (1 to 2 s) of combined steam (95°C) and ultrasound (SonoSteam) of industrial fish and meat transportation boxes and live-chicken transportation crates naturally contaminated with food and fecal residues were investigated. Aerobic counts of 5.0 to 6.0 log CFU/24 cm(2) and an Enterobacteriaceae spp. level of 2.0 CFU/24 cm(2) were found on the surfaces prior to the treatment. After 1 s of treatment, the aerobic counts were significantly (P < 0.0001) reduced, and within 2 s, reductions below the detection limit (<10 CFU) were reached. Enterobacteriaceae spp. were reduced to a level below the detection limit with only 1 s of treatment. Two seconds of steam-ultrasound treatment was also applied on two different types of plastic modular conveyor belts with hinge pins and one type of flat flexible rubber belt, all visibly contaminated with food residues. The aerobic counts of 3.0 to 5.0 CFU/50 cm(2) were significantly (P < 0.05) reduced, while Enterobacteriaceae spp. were reduced to a level below the detection limit. Industrial meat knives were contaminated with aerobic counts of 6.0 log CFU/5 cm(2) on the handle and 5.2 log CFU/14 cm(2) on the steel. The level of Enterobacteriaceae spp. contamination was approximately 2.5 log CFU on the handle and steel. Two seconds of steam-ultrasound treatment reduced the aerobic counts and Enterobacteriaceae spp. to levels below the detection limit on both handle and steel. This study shows that the steam-ultrasound treatment may be an effective replacement for disinfection processes and that it can be used for continuous disinfection at fast process lines. However, the treatment may not be able to replace efficient cleaning processes used to remove high loads of debris.

Comparison of cleaning efficacy between in-use disinfectant and electrolysed water in an English residential care home.

PubMed

Meakin, N S; Bowman, C; Lewis, M R; Dancer, S J

2012-02-01

Infection control in hospitals and care homes remains a key issue. They are regularly inspected regarding standards of hygiene, but visual assessment does not necessarily correlate with microbial cleanliness. Pathogens can persist in the inanimate environment for extended periods of time. This prospective study compared the effectiveness of a novel sanitizer containing electrolysed water, in which the active ingredient is stabilized hypochlorous acid (Aqualution™), with the effectiveness of the quaternary ammonium disinfectant in current use for microbial removal from hand-touch surfaces in a care home. The study had a two-period crossover design. Five surfaces were cleaned daily over a four-week period, with screening swabs taken before and after cleaning. Swabs were cultured in order to compare levels of surface microbial contamination [colony-forming units (cfu)/cm(2)] before and after cleaning with each product. Cleaning with electrolysed water reduced the mean surface bacterial load from 2.6 [interquartile range (IQR) 0.30-30.40] cfu/cm(2) to 0.10 (IQR 0.10-1.40) cfu/cm(2) [mean log(10) reduction factor 1.042, 95% confidence interval (CI) 0.79-1.30]. Cleaning with the in-use quaternary ammonium disinfectant increased the bacterial load from 0.90 (IQR 0.10-8.50) cfu/cm(2) to 93.30 (IQR 9.85-363.65) cfu/cm(2) (mean log(10) reduction -1.499, 95% CI -1.87 to -1.12) (P < 0.0001). Using two proposed benchmark standards for surface microbial levels in hospitals, electrolysed water resulted in a higher 'pass rate' than the in-use quaternary ammonium disinfectant (80-86% vs 15-21%, P < 0.0001). Electrolysed water exerts a more effective bacterial kill than the in-use quaternary ammonium disinfectant, which suggests that it may be useful as a surface sanitizer in environments such as care homes. Copyright © 2011 The Healthcare Infection Society. Published by Elsevier Ltd. All rights reserved.

An evaluation of selected methods for the decontamination of cattle hides prior to skinning.

PubMed

Small, A; Wells-Burr, B; Buncic, S

2005-02-01

The effectiveness of different decontamination treatments in reducing microbial loads on cattle hides was assessed. The 10-s hide treatments were conducted using a wet-and-dry vacuum cleaner filled with one of the liquids (heated to 50 °C) indicated below, followed or not by 10-min drying in the air. Also, the hide was clipped, followed or not by 10-s singeing using a hand-held blowtorch. Before and after each decontamination treatment, the hide was sampled (100 cm(2) areas) by a sponge-swabbing method to compare the total viable counts of bacteria (TVC). The largest bacterial reduction (P<0.001; 2.31log(10) cfu/cm(2)) was achieved by singeing of previously clipped hide. Treatment of hide with a food industry sanitizer solution (10% Betane Plus) resulted in significant reductions of 1.80 (P<0.001) and 1.98log(10) cfu/cm(2) (P<0.001) without and with subsequent drying, respectively. Treatment of hide with a food industry disinfectant (P3-Topactive DES) significantly reduced TVC by 0.97 (P<0.001) and 1.18log(10) cfu/cm(2) (P<0.001) without and with subsequent drying, respectively. Treatments of hide with water alone or with a food-safe detergent solution (Formula 963B), or hide clipping alone, did not produce significant decontamination effects. Since hide contamination is associated with microbial contamination of the carcasses, the results indicate that post-killing/pre-skinning hide decontamination (used alone, or in combination with carcass decontamination) has a potential to improve microbial meat safety. Nevertheless, further research is required to optimise the efficacy of these treatments in the reduction of specific pathogens under commercial conditions.

Potential of Lactobacillus curvatus LFC1 to produce slits in Cheddar cheese.

PubMed

Porcellato, D; Johnson, M E; Houck, K; Skeie, S B; Mills, D A; Kalanetra, K M; Steele, J L

2015-08-01

Defects in Cheddar cheese resulting from undesired gas production are a sporadic problem that results in significant financial losses in the cheese industry. In this study, we evaluate the potential of a facultatively heterofermentative lactobacilli, Lactobacillus curvatus LFC1, to produce slits, a gas related defect in Cheddar cheese. The addition of Lb. curvatus LFC1 to cheese milk at log 3 CFU/ml resulted in the development of small slits during the first month of ripening. Chemical analyses indicated that the LFC1 containing cheeses had less galactose and higher levels of lactate and acetate than the control cheeses. The composition the cheese microbiota was examined through a combination of two culture independent approaches, 16S rRNA marker gene sequencing and automated ribosomal intergenic spacer analysis; the results indicated that no known gas producers were present and that high levels of LFC1 was the only significant difference between the cheese microbiotas. A ripening cheese model system was utilized to examine the metabolism of LFC1 under conditions similar to those present in cheeses that exhibited the slit defect. The combined cheese and model system results indicate that when Lb. curvatus LFC1 was added to the cheese milk at log 3 CFU/ml it metabolized galactose to lactate, acetate, and CO2. For production of sufficient CO2 to result in the formation of slits there needs to be sufficient galactose and Lb. curvatus LFC1 present in the cheese matrix. To our knowledge, facultatively heterofermentative lactobacilli have not previously been demonstrated to result in gas-related cheese defects. Copyright © 2015 Elsevier Ltd. All rights reserved.

Rain Splash Dispersal of Gibberella zeae Within Wheat Canopies in Ohio.

PubMed

Paul, P A; El-Allaf, S M; Lipps, P E; Madden, L V

2004-12-01

ABSTRACT Rain splash dispersal of Gibberella zeae, causal agent of Fusarium head blight of wheat, was investigated in field studies in Ohio between 2001 and 2003. Samplers placed at 0, 30, and 100 cm above the soil surface were used to collect rain splash in wheat fields with maize residue on the surface and fields with G. zeae-infested maize kernels. Rain splash was collected during separate rain episodes throughout the wheat-growing seasons. Aliquots of splashed rain were transferred to petri dishes containing Komada's selective medium, and G. zeae was identified based on colony and spore morphology. Dispersed spores were measured in CFU/ml. Intensity of splashed rain was highest at 100 cm and ranged from 0.2 to 10.2 mm h(-1), depending on incident rain intensity and sampler height. Spores were recovered from splash samples at all heights in both locations for all sampled rain events. Both macroconidia and ascospores were found based on microscopic examination of random samples of splashed rain. Spore density and spore flux density per rain episode ranged from 0.4 to 40.9 CFU cm(-2) and 0.4 to 84.8 CFU cm(-2) h(-1), respectively. Spore flux density was higher in fields with G. zeae-infested maize kernels than in fields with maize debris, and generally was higher at 0 and 30 cm than at 100 cm at both locations. However, on average, spore flux density was only 30% lower at 100 cm (height of wheat spikes) than at the other heights. The log of spore flux density was linearly related to the log of splashed rain intensity and the log of incident rain intensity. The regression slopes were not significantly affected by year, location, height, and their interactions, but the intercepts were significantly affected by both sampler height and location. Thus, our results show that spores of G. zeae were consistently splash dispersed to spike heights within wheat canopies, and splashed rain intensity and spore flux density could be predicted based on incident rain intensity in order to estimate inoculum dispersal within the wheat canopy.

Growth of Salmonella on sprouting alfalfa seeds as affected by the inoculum size, native microbial load and Pseudomonas fluorescens 2-79.

PubMed

Liao, C-H

2008-02-01

To investigate the growth of salmonellae on sprouting alfalfa seeds as affected by the inoculum size, microbial load and Pseudomonas fluorescens 2-79. Alfalfa seeds pre-inoculated with < or =10(1)-10(3) CFU g(-1) of salmonellae and with or without Ps. fluorescens 2-79 were sprouted in glass jars and the population of salmonellae were determined daily for up to 6 days. The population of salmonellae on germinating seeds reached the maximum 2-3 days after sprouting when total bacterial count reached the maximum (10(9) CFU g(-1)). The population of salmonellae on sprouting seeds not treated with Ps. fluorescens 2-79 showed a net increase of 3-4 log units. However, the population of salmonellae on alfalfa seeds treated with Ps. fluorescens 2-79 showed a net increase of only 1-2 log units. Disinfection of seeds with calcium hypochlorite enhanced the growth of salmonellae. Treatment of seeds with Ps. fluorescens 2-79 reduced the growth of salmonellae by 2-3 log units. The potential of Ps. fluorescens 2-79 as a biological agent for use in control of salmonellae on sprouting seeds was demonstrated and warrants further investigation.

Salmonella Typhimurium and Salmonella Sofia: Growth in and Persistence on Eggs under Production and Retail Conditions

PubMed Central

McAuley, Catherine M.; Duffy, Lesley L.; Subasinghe, Nela; Hogg, Geoff; Coventry, John; Fegan, Narelle

2015-01-01

Salmonellosis in Australia has been linked to eggs and egg products with specific serotypes associated with outbreaks. We compared attachment to and survival on egg shells and growth in eggs of two Salmonella serotypes, an egg outbreak associated Salmonella Typhimurium and a non-egg-associated Salmonella enterica ssp. II 1,4,12,27:b:[e,n,x] (S. Sofia). Experiments were conducted at combinations of 4, 15, 22, 37 and 42°C. No significant differences occurred between the serotypes in maximum growth rates, which were significantly greater (P < 0.001) in egg yolk (0.427 log10 CFU/mL/h) compared to whole egg (0.312 log10 CFU/mL/h) and egg white (0.029 log10 CFU/mL/h). Attachment to egg shells varied by time (1 or 20 min) and temperature (4, 22 and 42°C), with S. Typhimurium isolates attaching at higher levels (P < 0.05) than S. Sofia after 1 min at 4°C and S. Typhimurium ATCC 14028 attaching at higher (P < 0.05) levels at 22°C. Survival on egg shells was not significantly different across isolates. Salmonella serotypes behaved similarly regarding growth in egg contents, attachment to egg shells and survival on eggs, indicating that other factors more likely contributed to reasons for S. Typhimurium being implicated in multiple egg-associated outbreaks. PMID:26539536

Study of the quality characteristics in cold-smoked salmon (Salmo salar) originating from pre- or post-rigor raw material.

PubMed

Birkeland, S; Akse, L

2010-01-01

Improved slaughtering procedures in the salmon industry have caused a delayed onset of rigor mortis and, thus, a potential for pre-rigor secondary processing. The aim of this study was to investigate the effect of rigor status at time of processing on quality traits color, texture, sensory, microbiological, in injection salted, and cold-smoked Atlantic salmon (Salmo salar). Injection of pre-rigor fillets caused a significant (P<0.001) contraction (-7.9%± 0.9%) on the caudal-cranial axis. No significant differences in instrumental color (a*, b*, C*, or h*), texture (hardness), or sensory traits (aroma, color, taste, and texture) were observed between pre- or post-rigor processed fillets; however, post-rigor (1477 ± 38 g) fillets had a significant (P>0.05) higher fracturability than pre-rigor fillets (1369 ± 71 g). Pre-rigor fillets were significantly (P<0.01) lighter, L*, (39.7 ± 1.0) than post-rigor fillets (37.8 ± 0.8) and had significantly lower (P<0.05) aerobic plate count (APC), 1.4 ± 0.4 log CFU/g against 2.6 ± 0.6 log CFU/g, and psychrotrophic count (PC), 2.1 ± 0.2 log CFU/g against 3.0 ± 0.5 log CFU/g, than post-rigor processed fillets. This study showed that similar quality characteristics can be obtained in cold-smoked products processed either pre- or post-rigor when using suitable injection salting protocols and smoking techniques. © 2010 Institute of Food Technologists®

Effect of packaging and storage time on survival of Listeria monocytogenes on kippered beef steak and turkey tenders.

PubMed

Uppal, Kamaldeep K; Getty, Kelly J K; Boyle, Elizabeth A E; Harper, Nigel M; Lobaton-Sulabo, April Shayne S; Barry, Bruce

2012-01-01

The objective of our study was to determine effect of packaging method and storage time on reducing Listeria monocytogenes in shelf-stable meat snacks. Commercially available kippered beef steak strips and turkey tenders were dipped into a 5-strain L. monocytogenes cocktail, and dried at 23 °C until a water activity of 0.80 was achieved. Inoculated samples were packaged with 4 treatments: (1) vacuum, (2) nitrogen flushed with oxygen scavenger, (3) heat sealed with oxygen scavenger, and (4) heat sealed without oxygen scavenger. Samples were stored at 23 °C and evaluated for L. monocytogenes levels at 0, 24, 48, and 72 h. Initial levels (time 0) of L. monocytogenes were approximately 5.7 log CFU/cm² for steak and tenders. After 24 h of storage time, a 1 log CFU/cm² reduction of L. monocytogenes was observed for turkey tenders for all packaging treatments. After 48 h, turkey tenders showed >1 log CFU/cm² reduction of L. monocytogenes for all packaging treatments except for vacuum, where only 0.9 log CFU/cm² reduction was observed. After 72 h, reductions for all packaging treatments for turkey tenders ranged from 1.5 to 2.4 log CFU/cm². For kippered beef steak, there was no interaction between the packaging treatments and all storage times (P > 0.05) whereas, time was different (P <0.05). For kippered beef steak, there was 1 log reduction of L. monocytogenes at 24 and 48 h of storage times at 23 °C for all packaging treatments and a 2.1 log CFU/ cm² L. monocytogenes reduction at 72 h of storage time. Processors of kippered beef steak and turkey tenders could use a combination of vacuum or nitrogen-flushing or heat sealed with an oxygen scavenger packaging methods and a holding time of 24 h prior to shipping to reduce potential L. monocytogenes numbers by ≥1 log. However, processors should be encouraged to hold packaged product a minimum of 72 h to enhance the margin of safety for L. monocytogenes control. © 2011 Institute of Food Technologists®

The effect of short G-CSF administration on the numbers and clonogenic efficiency of hematopoietic progenitor cells in bone marrow and peripheral blood of normal donors.

PubMed

Zaucha, J M; Knopińska-Posłuszny, W; Bieniaszewska, M; Myśliwski, A; Hellmann, A

2000-01-01

We have analysed the cellularity, the number of clonogenic cells and their clonogenic efficiency (the number of clonogenic cells/2 x 10(5) MNC) in peripheral blood (PB) and bone marrow (BM) during and after filgrastim (rhG-CSF) mobilization of CD34+ cells in 12 healthy donors for allogeneic stem cell donation. G-CSF was administrated subcutaneously for 5 consecutive days at a dose of 10 micrograms/kg/day. WBC, MNC, CD34+ cell counts, CFU-GM and BFU-E assays in PB were performed at baseline and then daily 12 hours after each G-CSF dose. BM was assayed before start (day 1) and after the last dose (day 6) of G-CSF. Results are given as medians, with ranges in parentheses. In PB the total WBC and MNC increased 7.4-fold (6.0-12.3) and 3.3-fold (1.5-9.4), respectively, reaching a peak of 49.4 x 10(9)/l (32.5-66.6) on day 6 for WBC and 6.28 x 10(9)/l (4.7-13.3) for MNC on day 5. CD34+ cell number reached a peak value of 48.0 x 10(6)/l (45.6-285) on day 6 whereas CFU-GM and BFU-E reached their peaks on day 5, 0.95 x 10(4)/ml (0.05-6.08) and 1.04 x 10(4)/ml, respectively. CFU-MIX, not detectable at baseline, reached a peak of 0.95 x 10(4)/ml (0.006-0.51) on day 5 as well. This was accompanied by an increase in CFU-GM, BFU-E and CFU-MIX clonogenic efficiency: 23-fold (3-150), 9.75-fold (2.2-27.8) and 20-fold (2.5-210), respectively. In BM the total WBC number increased 2.5-fold (1.3-4.9) from the baseline value of 52.6 x 10(9)/l (7.9-137.0) whereas the MNC count increased 2.0-fold (0.81-3.7) from a baseline of 13.6 x 10(9)/l (3.5-54.8). This was, however, not significant. The number of CD34+ cells increased significantly 2.9-fold (0.8-8.3). In 8 donors CFU-MIX were detectable before but not after G-CSF treatment. A similar decrease in CFU-GM and BFU-E clonogenic efficiency occurred but was not significant. CFU-GM and BFU-E numbers did not change. We conclude that the total body numbers of lineage committed progenitors increased during G-CSF administration, which indicate their proliferation in addition to mobilization. The effect of G-CSF on the number of more primitive progenitors in BM is less clear and needs further investigation.

Preslaughter diet management in sheep and goats: effects on physiological responses and microbial loads on skin and carcass

PubMed Central

2014-01-01

Sixteen crossbred buck goats (Kiko x Spanish; BW = 32.8 kg) and wether sheep (Dorset x Suffolk; BW = 39.9 kg) were used to determine the effect of preslaughter diet and feed deprivation time (FDT) on physiological responses and microbial loads on skin and carcasses. Experimental animals were fed either a concentrate (CD) or a hay diet (HD) for 4 d and then deprived of feed for either 12-h or 24-h before slaughter. Blood samples were collected for plasma cortisol and blood metabolite analyses. Longisimus muscle (LM) pH was measured. Skin and carcass swabs were obtained to assess microbial loads. Plasma creatine kinase activity (863.9 and 571.7 ± 95.21 IU) and non-esterified fatty acid concentrations (1,056.1 and 589.8 ± 105.01 mEq/L) were different (P < 0.05) between sheep and goats. Species and diet treatments had significant effects on the ultimate pH of LM. Pre-holding total coliform (TCC) and aerobic plate counts (APC) of skin were significantly different between species. Goats had lower (P < 0.05) TCC (2.1 vs. 3.0 log10 CFU/cm2) and APC (8.2 vs. 8.5 log10 CFU/cm2) counts in the skin compared to sheep. Preslaughter skin E. coli counts and TCC were different (P < 0.05) between species. Goats had lower (P < 0.05) counts of E. coli (2.2 vs. 2.9 log10 CFU/cm2) and TCC (2.3 vs. 3.0 log10 CFU/cm2) in the skin compared with those in sheep. Diet, species, and FDT had no effect (P > 0.05) on E. coli and TCC in carcass swab samples. The APC of carcass swab samples were only affected (P < 0.05) by the FDT. The results indicated that preslaughter dietary management had no significant changes on hormone and blood metabolite concentrations and sheep might be more prone for fecal contamination than goats in the holding pens at abattoir. PMID:25343027

Reduction of Escherichia coli O157:H7 in Biofilms Using Bacteriophage BPECO 19.

PubMed

Sadekuzzaman, Mohammad; Yang, Sungdae; Mizan, Md Furkanur Rahaman; Ha, Sang-Do

2017-06-01

Biofilm formation is a growing concern in the food industry. Escherichia coli O157:H7 is one of the most important foodborne pathogens that can persists in food and food-related environments and subsequently produce biofilms. The efficacy of bacteriophage BPECO 19 was evaluated against three E. coli O157:H7 strains in biofilms. Biofilms of the three E. coli O157:H7 strains were grown on abiotic (stainless steel, rubber, and minimum biofilm eradication concentration [MBEC TM ] device) and biotic (lettuce) surfaces at different temperatures. The effectiveness of bacteriophage BPECO 19 in reducing preformed biofilms on these surfaces was further evaluated by treating the surfaces with a phage suspension (10 8 PFU/mL) for 2 h. The results indicated that the phage treatment significantly reduced (P  < 0.05) the number of adhered cells in all the surfaces. Following phage treatment, the viability of adhered cells was reduced by ≥3 log CFU/cm 2 , 2.4 log CFU/cm 2 , and 3.1 log CFU/peg in biofilms grown on stainless steel, rubber, and the MBEC TM device, respectively. Likewise, the phage treatment reduced cell viability by ≥2 log CFU/cm 2 in biofilms grown on lettuce. Overall, these results suggested that bacteriophages such as BPECO 19 could be effective in reducing the viability of biofilm-adhered cells. © 2017 Institute of Food Technologists®.

Antimicrobial efficacy of 3 oral antiseptics containing octenidine, polyhexamethylene biguanide, or Citroxx: can chlorhexidine be replaced?

PubMed

Rohrer, Nadine; Widmer, Andreas F; Waltimo, Tuomas; Kulik, Eva M; Weiger, Roland; Filipuzzi-Jenny, Elisabeth; Walter, Clemens

2010-07-01

Use of oral antiseptics decreases the bacterial load in the oral cavity. To compare the antimicrobial activity of 3 novel oral antiseptics with that of chlorhexidine, which is considered the "gold standard" of oral hygiene. Comparative in vitro study. Four common oral microorganisms (Streptococcus sanguinis, Streptococcus mutans, Candida albicans, and Fusobacterium nucleatum) were tested under standard conditions and at different concentrations, by use of a broth dilution assay and an agar diffusion assay and by calculating the log10 reduction factor (RF). The antimicrobial activity of each antiseptic was assessed by counting the difference in bacterial densities (ie, the log10 number of colony-forming units of bacteria) before and after the disinfection process. The oral antiseptics containing octenidine (with an RF in the range of 7.1-8.24 CFU/mL) and polyhexamethylene biguanide (with an RF in the range of 7.1-8.24 CFU/mL) demonstrated antimicrobial activity comparable to that of chlorhexidine (with an RF in the range of 1.03-8.24 CFU/mL), whereas the mouth rinse containing Citroxx (Citroxx Biosciences; with an RF in the range of 0.22-1.36 CFU/mL) showed significantly weaker antimicrobial efficacy. Overall, octenidine and polyhexamethylene biguanide were more active at lower concentrations.conclusion. Oral antiseptics containing the antimicrobial agent octenidine or polyhexamethylene biguanide may be considered as potent alternatives to chlorhexidine-based preparations.

Multilaboratory comparison of hepatitis C virus viral load assays.

PubMed

Caliendo, A M; Valsamakis, A; Zhou, Y; Yen-Lieberman, B; Andersen, J; Young, S; Ferreira-Gonzalez, A; Tsongalis, G J; Pyles, R; Bremer, J W; Lurain, N S

2006-05-01

We report a multilaboratory evaluation of hepatitis C virus (HCV) viral load assays to determine their linear range, reproducibility, subtype detection, and agreement. A panel of HCV RNA samples ranging in nominal concentration from 1.0 to 7.0 log10 IU/ml was constructed by diluting a clinical specimen (genotype 1b). Replicates of the panel were tested in multiple laboratories using the Abbott TaqMan analyte-specific reagent (Abbott reverse transcription-PCR [RT-PCR]), Roche TaqMan RUO (Roche RT-PCR), Roche Amplicor Monitor HCV 2.0 (Roche Monitor), and Bayer VERSANT HCV RNA 3.0 (Bayer bDNA) assays. Bayer bDNA-negative specimens were tested reflexively using the Bayer VERSANT HCV RNA qualitative assay (Bayer TMA). Abbott RT-PCR and Roche RT-PCR detected all 28 replicates with a concentration of 1.0 log10 IU/ml and were linear to 7.0 log10 IU/ml. Roche Monitor and Bayer bDNA detected 27 out of 28 and 13 out of 28 replicates, respectively, of 3.0 log10 IU/ml. Bayer TMA detected all seven replicates with 1.0 log10 IU/ml. Bayer bDNA was the most reproducible of the four assays. The mean viral load values for panel members in the linear ranges of the assays were within 0.5 log10 for the different tests. Eighty-nine clinical specimens of various genotypes (1 through 4) were tested in the Bayer bDNA, Abbott RT-PCR, and Roche RT-PCR assays. For Abbott RT-PCR, mean viral load values were 0.61 to 0.96 log10 greater than the values for Bayer bDNA assay for samples with genotype 1, 2, or 3 samples and 0.08 log10 greater for genotype 4 specimens. The Roche RT-PCR assay gave mean viral load values that were 0.28 to 0.82 log10 greater than those obtained with the Bayer bDNA assay for genotype 1, 2, and 3 samples. However, for genotype 4 samples the mean viral load value obtained with the Roche RT-PCR assay was, on average, 0.15 log10 lower than that of the Bayer bDNA. Based on these data, we conclude that the sensitivity and linear range of the Abbott and Roche RT-PCR assays enable them to be used for HCV diagnostics and therapeutic monitoring. However, the differences in the viral load values obtained with the different assays underscore the importance of using one assay when monitoring response to therapy.

Pharmacodynamic Evaluation of the Intracellular Activity of Antibiotics towards Pseudomonas aeruginosa PAO1 in a Model of THP-1 Human Monocytes

PubMed Central

Buyck, Julien M.; Tulkens, Paul M.

2013-01-01

Pseudomonas aeruginosa invades epithelial and phagocytic cells, which may play an important role in the persistence of infection. We have developed a 24-h model of THP-1 monocyte infection with P. aeruginosa PAO1 in which bacteria are seen multiplying in vacuoles by electron microscopy. The model has been used to quantitatively assess antibiotic activity against intracellular and extracellular bacteria by using a pharmacodynamic approach (concentration-dependent experiments over a wide range of extracellular concentrations to calculate bacteriostatic concentrations [Cs] and maximal relative efficacies [Emax]; Hill-Langmuir equation). Using 16 antipseudomonal antibiotics (three aminoglycosides, nine β-lactams, three fluoroquinolones, and colistin), dose-response curves were found to be undistinguishable for antibiotics of the same pharmacological class if data were expressed as a function of the corresponding MICs. Extracellularly, all of the antibiotics reached a bacteriostatic effect at their MIC, and their Emax exceeded the limit of detection (−4.5 log10 CFU compared to the initial inoculum). Intracellularly, Cs values remained unchanged for β-lactams, fluoroquinolones, and colistin but were approximately 10 times higher for aminoglycosides, whereas Emax values were markedly reduced (less negative), reaching −3 log10 CFU for fluoroquinolones and only −1 to −1.5 log10 CFU for all other antibiotics. The decrease in intracellular aminoglycoside potency (higher Cs) can be ascribed to the acid pH to which bacteria are exposed in vacuoles. The decrease in the Emax may reflect a reversible alteration of bacterial responsiveness to antibiotics in the intracellular milieu. The model may prove useful for comparison of antipseudomonal antibiotics to reduce the risk of persistence or relapse of pseudomonal infections. PMID:23478951

Cold plasma rapid decontamination of food contact surfaces contaminated with Salmonella biofilms.

PubMed

Niemira, Brendan A; Boyd, Glenn; Sites, Joseph

2014-05-01

Cross-contamination of foods from persistent pathogen reservoirs is a known risk factor in processing environments. Industry requires a rapid, waterless, zero-contact, chemical-free method for removing pathogens from food contact surfaces. Cold plasma was tested for its ability to inactivate Salmonella biofilms. A 3-strain Salmonella culture was grown to form adherent biofilms for 24, 48, or 72 h on a test surface (glass slides). These were placed on a conveyor belt and passed at various line speeds to provide exposure times of 5, 10, or 15 s. The test plate was either 5 or 7.5 cm under a plasma jet emitter operating at 1 atm using filtered air as the feed gas. The frequency of high-voltage electricity was varied from 23 to 48 kHz. At the closer spacing (5 cm), cold plasma reduced Salmonella biofilms by up to 1.57 log CFU/mL (5 s), 1.82 log CFU/mL (10 s), and 2.13 log CFU/mL (15 s). Increasing the distance to 7.5 cm generally reduced the efficacy of the 15 s treatment, but had variable effects on the 5 and 10 s treatments. Variation of the high-voltage electricity had a greater effect on 10 and 15 s treatments, particularly at the 7.5 cm spacing. For each combination of time, distance, and frequency, Salmonella biofilms of 24, 48, and 72 h growth responded consistently with each other. The results show that short treatments with cold plasma yielded up to a 2.13 log reduction of a durable form of Salmonella contamination on a model food contact surface. This technology shows promise as a possible tool for rapid disinfection of materials associated with food processing. Pathogens such as Salmonella can form chemical-resistant biofilms, making them difficult to remove from food contact surfaces. A 15 s treatment with cold plasma reduced mature Salmonella biofilms by up to 2.13 log CFU/mL (99.3%). This contact-free, waterless method uses no chemical sanitizers. Cold plasma may therefore have a practical application for conveyor belts, equipment, and other food contact surfaces where a rapid, dry antimicrobial process is required. © 2014 Institute of Food Technologists®

Efficacy of a Fixed Combination of Tetracycline, Chloramphenicol, and Colistimethate Sodium for Treatment of Candida albicans Keratitis.

PubMed

Blanco, Anna R; Nostro, Antonia; D'Angelo, Valeria; D'Arrigo, Manuela; Mazzone, Maria G; Marino, Andreana

2017-08-01

To evaluate the antifungal activity of a fixed antibiotic combination (AC) containing tetracycline (TET), chloramphenicol (CAF), and colistimethate sodium (CS). In vitro: Candida ATCC and clinical strains were used. The minimum inhibitory concentrations (MICs) of AC and of each antibiotic were determined. Fluconazole (FLC) was tested for comparison. Time-killing curves of selected strains were performed. Ex vivo keratitis: corneas were injected intrastromally with the selected strains. After the injection, corneas were divided into groups of treatments: AC, FLC, or saline. Then, the tissues were analyzed for colony-forming units per gram (CFU/g). Propidium iodide (PI) and MitoTracker (MTR) staining were used to investigate the mode of action. Values of MIC required to inhibit the growth of 90% of organisms for the antibiotics alone were higher than FLC. However, their activity was enhanced when used in combination against Candida yeasts. Time-killing curves showed that at 24 hours, AC reduced the load of both strains of approximately 1 Log10 CFU/g compared with the initial inoculum (P < 0.0001). This effect was also significant versus FLC. In ex vivo, AC was effective in decreasing the loads of both strains by 4 Log10 CFU/g with respect to the control. Moreover, it showed higher activity than FLC against Candida albicans ATCC 10231 (1 Log10 CFU/g, P < 0.01 versus control). PI staining demonstrated that CS changed the membrane's permeability, whereas MTR staining demonstrated that TET or CAF altered mitochondrial function. The cells treated with AC and stained showed both effects. In this study, AC showed antifungal efficacy versus Candida spp.; this activity can be due to the synergistic effects of antibiotics in it.

Adhesion of Porphyromonas gingivalis and Tannerella forsythia to dentin and titanium with sandblasted and acid etched surface coated with serum and serum proteins - An in vitro study.

PubMed

Eick, Sigrun; Kindblom, Christian; Mizgalska, Danuta; Magdoń, Anna; Jurczyk, Karolina; Sculean, Anton; Stavropoulos, Andreas

2017-03-01

To evaluate the adhesion of selected bacterial strains incl. expression of important virulence factors at dentin and titanium SLA surfaces coated with layers of serum proteins. Dentin- and moderately rough SLA titanium-discs were coated overnight with human serum, or IgG, or human serum albumin (HSA). Thereafter, Porphyromonas gingivalis, Tannerella forsythia, or a six-species mixture were added for 4h and 24h. The number of adhered bacteria (colony forming units; CFU) was determined. Arg-gingipain activity of P. gingivalis and mRNA expressions of P. gingivalis and T. forsythia proteases and T. forsythia protease inhibitor were measured. Coating specimens never resulted in differences exceeding 1.1 log10 CFU, comparing to controls, irrespective the substrate. Counts of T. forsythia were statistically significantly higher at titanium than dentin, the difference was up to 3.7 log10 CFU after 24h (p=0.002). No statistically significant variation regarding adhesion of the mixed culture was detected between surfaces or among coatings. Arg-gingipain activity of P. gingivalis was associated with log10 CFU but not with the surface or the coating. Titanium negatively influenced mRNA expression of T. forsythia protease inhibitor at 24h (p=0.026 uncoated, p=0.009 with serum). The present findings indicate that: a) single bacterial species (T. forsythia) can adhere more readily to titanium SLA than to dentin, b) low expression of T. forsythia protease inhibitor may influence the virulence of the species on titanium SLA surfaces in comparison with teeth, and c) surface properties (e.g. material and/or protein layers) do not appear to significantly influence multi-species adhesion. Copyright © 2016 Elsevier Ltd. All rights reserved.

Growth and survival of Escherichia coli O157:H7 and Listeria monocytogenes in egg products held at different temperatures.

PubMed

Yang, S E; Chou, C C

2000-07-01

Growth and survival of Escherichia coli O157:H7 and Listeria monocytogenes in steamed eggs and scrambled eggs held at different temperatures (5, 18, 22, 37, 55, and 60 degrees C) were investigated in the present study. Among the holding temperatures tested, both pathogens multiplied best at 37 degrees C followed by 22, 18, and 5 degrees C. In general, E. coli O157:H7 grew better in the egg products than L. monocytogenes did at all the storage temperatures tested except at 5 degrees C. E. coli O157:H7 did not grow in steamed eggs and scrambled eggs held at 5 degrees C. L. monocytogenes showed a slight population increase of approximately 0.6 to 0.9 log CFU/g in these egg products at the end of the 36-h storage period at 5 degrees C. The population of both pathogens detected in the egg products was affected by the initial population, holding temperature, and length of the holding period. It was also noted that L. monocytogenes was more susceptible than E. coli O157:H7 in steamed eggs held at 60 degrees C. After holding at 60 degrees C for 1 h, no detectable viable cells of L. monocytogenes with a population reduction of 5.4 log CFU/g was observed in steamed eggs, whereas a lower population reduction of only approximately 0.5 log CFU/ml was noted for E. coli O157:H7.

Efficacy of two hydrogen peroxide vapour aerial decontamination systems for enhanced disinfection of meticillin-resistant Staphylococcus aureus, Klebsiella pneumoniae and Clostridium difficile in single isolation rooms.

PubMed

Ali, S; Muzslay, M; Bruce, M; Jeanes, A; Moore, G; Wilson, A P R

2016-05-01

Hydrogen peroxide vapour (HPV) disinfection systems are being used to reduce patients' exposure to hospital pathogens in the environment. HPV whole-room aerial disinfection systems may vary in terms of operating concentration and mode of delivery. To assess the efficacy of two HPV systems (HPS1 and HPS2) for whole-room aerial disinfection of single isolation rooms (SIRs). Ten SIRs were selected for manual terminal disinfection after patient discharge. Test coupons seeded with biological indicator (BI) organisms [∼10(6) colony-forming units (cfu) of meticillin-resistant Staphylococcus aureus (MRSA) or Klebsiella pneumoniae, or ∼10(5)cfu Clostridium difficile 027 spores] prepared in a soil challenge were placed at five locations per room. For each cycle, 22 high-frequency-touch surfaces in SIRs were sampled with contact plates (∼25cm(2)) before and after HPV decontamination, and BIs were assayed for the persistence of pathogens. Approximately 95% of 214 sites were contaminated with bacteria after manual terminal disinfection, with high numbers present on the SIR floor (238.0-352.5cfu), bed control panel (24.0-33.5cfu), and nurse call button (21.5-7.0cfu). Enhanced disinfection using HPV reduced surface contamination to low levels: HPS1 [0.25cfu, interquartile range (IQR) 0-1.13] and HPS2 (0.5cfu, IQR 0-2.0). Both systems demonstrated similar turnaround times (∼2-2.5h), and no differences were observed in the efficacy of the two systems against BIs (C. difficile ∼5.1log10 reduction; MRSA/K. pneumoniae ∼6.3log10 reduction). Despite different operating concentrations of hydrogen peroxide, MRSA persisted on 27% of coupons after HPV decontamination. Enhanced disinfection with HPV reduces surface contamination left by manual terminal cleaning, minimizing the risks of cross-contamination. The starting concentration and mode of delivery of hydrogen peroxide may not improve the efficacy of decontamination in practice, and therefore the choice of HPV system may be based upon other considerations such as cost, convenience and logistics. Copyright © 2016. Published by Elsevier Ltd.

Bacteriophage biocontrol of Listeria monocytogenes on soft ripened white mold and red-smear cheeses.

PubMed

Guenther, Susanne; Loessner, Martin J

2011-03-01

Soft-ripened cheeses belong to the type of food most often contaminated with Listeria monocytogenes, and they have been implicated in several outbreaks of listeriosis. Bacteriophages represent an attractive way to combat foodborne pathogens without affecting other properties of the food. We used the broad host range, virulent Listeria phage A511 for control of L. monocytogenes during the production and ripening phases of both types of soft-ripened cheeses, white mold (Camembert-type) cheese, as well as washed-rind cheese with a red-smear surface (Limburger-type). The surfaces of young, unripened cheese were inoculated with 10(1)-10(3) cfu/cm(2)L. monocytogenes strains Scott A (serovar 4b) or CNL 10(3)/2005 (serovar 1/2a). Phage was applied at defined time points thereafter, in single or repeated treatments, at 3 × 10(8) or 1 × 10(9) pfu/cm(2). With Scott A (10(3) cfu/cm(2)) and a single dose of A511 (3 × 10(8) pfu/cm(2)) on camembert-type cheese, viable counts dropped 2.5 logs at the end of the 21 day ripening period. Repeated phage application did not further inhibit the bacteria, whereas a single higher dose (1 × 10(9) pfu/cm(2)) was found to be more effective. On red-smear cheese ripened for 22 days, Listeria counts were down by more than 3 logs. Repeated application of A511 further delayed re-growth of Listeria, but did not affect bacterial counts after 22 days. With lower initial Listeria contamination (10(1)-10(2) cfu/cm(2)), viable counts dropped below the limit of detection, corresponding to more than 6 logs reduction compared to the control. Our data clearly demonstrate the potential of bacteriophage for biocontrol of L. monocytogenes in soft cheese.

Efficacy of ultraviolet light exposure against survival of Listeria monocytogenes on conveyor belts.

PubMed

Morey, Amit; McKee, Shelly R; Dickson, James S; Singh, Manpreet

2010-06-01

Listeria monocytogenes has been repeatedly isolated from foods and food-processing facilities including food contact surfaces such as conveyor belts (CB). CBs are often difficult to clean and require rigorous sanitation programs for decontamination. Ultraviolet (UV) light has exhibited microbicidal properties on food contact surfaces and this study was conducted to determine the efficacy of UV against L. monocytogenes on CB made of different materials. A four-strain cocktail of L. monocytogenes (serotypes 3A, 4A, 4B, and 4C) was made to give a suspension of approximately 10(7) CFU/mL. CBs made from four different types of materials, (1) Ropanyl DM 8/2 A2 + 04 (belt 1), (2) Volta FRMW-3.0 (belt 2), (3) Volta FRMB-3.0 (belt 3), and (4) Ropanyl DM (belt 4), were inoculated with 1 mL of the four-strain cocktail (approximately 10(7) CFU/mL) of the bacterial suspension. CBs were treated with UV light (254 nm) for 1 and 3 sec at 5.53 and 5.95 mW/cm(2). Three replications of the experiments were conducted. Two-way analysis of variance of survival populations of L. monocytogenes showed that bacterial counts were significantly reduced (p < 0.05) on all belt types irrespective of UV light intensities and times of exposure. L. monocytogenes populations were reduced (p < 0.05) to below detection limits on belts 1, 2, and 3 after exposure to 5.95 mW/cm(2) UV light intensity for 3 sec. L. monocytogenes-inoculated CBs that were exposed to 5.53 mW/cm(2) showed higher (p < 0.05) survival populations of L. monocytogenes compared with 5.95 mW/cm(2) on all the four CBs. Belt 4 showed survival populations of L. monocytogenes ranging from 1.42 to 1.73 log(10) CFU/cm(2) after UV light treatment for 1 and 3 sec. UV light can be effectively used to reduce L. monocytogenes contamination on CBs.

Intracellular activity of antibiotics in a model of human THP-1 macrophages infected by a Staphylococcus aureus small-colony variant strain isolated from a cystic fibrosis patient: pharmacodynamic evaluation and comparison with isogenic normal-phenotype and revertant strains.

PubMed

Nguyen, Hoang Anh; Denis, Olivier; Vergison, Anne; Theunis, Anne; Tulkens, Paul M; Struelens, Marc J; Van Bambeke, Françoise

2009-04-01

Small-colony variant (SCV) strains of Staphylococcus aureus show reduced antibiotic susceptibility and intracellular persistence, potentially explaining therapeutic failures. The activities of oxacillin, fusidic acid, clindamycin, gentamicin, rifampin, vancomycin, linezolid, quinupristin-dalfopristin, daptomycin, tigecycline, moxifloxacin, telavancin, and oritavancin have been examined in THP-1 macrophages infected by a stable thymidine-dependent SCV strain in comparison with normal-phenotype and revertant isogenic strains isolated from the same cystic fibrosis patient. The SCV strain grew slowly extracellularly and intracellularly (1- and 0.2-log CFU increase in 24 h, respectively). In confocal and electron microscopy, SCV and the normal-phenotype bacteria remain confined in acid vacuoles. All antibiotics tested, except tigecycline, caused a net reduction in bacterial counts that was both time and concentration dependent. At an extracellular concentration corresponding to the maximum concentration in human serum (total drug), oritavancin caused a 2-log CFU reduction at 24 h; rifampin, moxifloxacin, and quinupristin-dalfopristin caused a similar reduction at 72 h; and all other antibiotics had only a static effect at 24 h and a 1-log CFU reduction at 72 h. In concentration dependence experiments, response to oritavancin was bimodal (two successive plateaus of -0.4 and -3.1 log CFU); tigecycline, moxifloxacin, and rifampin showed maximal effects of -1.1 to -1.7 log CFU; and the other antibiotics produced results of -0.6 log CFU or less. Addition of thymidine restored intracellular growth of the SCV strain but did not modify the activity of antibiotics (except quinupristin-dalfopristin). All drugs (except tigecycline and oritavancin) showed higher intracellular activity against normal or revertant phenotypes than against SCV strains. The data may help rationalizing the design of further studies with intracellular SCV strains.

Rapid Assessment of the Viability of Mycobacterium avium subsp. paratuberculosis Cells after Heat Treatment, Using an Optimized Phage Amplification Assay▿

PubMed Central

Foddai, Antonio; Elliott, Christopher T.; Grant, Irene R.

2010-01-01

Thermal inactivation experiments were carried out to assess the utility of a recently optimized phage amplification assay to accurately enumerate viable Mycobacterium avium subsp. paratuberculosis cells in milk. Ultra-heat-treated (UHT) whole milk was spiked with large numbers of M. avium subsp. paratuberculosis organisms (106 to 107 CFU/ml) and dispensed in 100-μl aliquots in thin-walled 200-μl PCR tubes. A Primus 96 advanced thermal cycler (Peqlab, Erlangen, Germany) was used to achieve the following time and temperature treatments: (i) 63°C for 3, 6, and 9 min; (ii) 68°C for 20, 40, and 60 s; and (iii) 72°C for 5, 10, 15, and 25 s. After thermal stress, the number of surviving M. avium subsp. paratuberculosis cells was assessed by both phage amplification assay and culture on Herrold's egg yolk medium (HEYM). A high correlation between PFU/ml and CFU/ml counts was observed for both unheated (r2 = 0.943) and heated (r2 = 0.971) M. avium subsp. paratuberculosis cells. D and z values obtained using the two types of counts were not significantly different (P > 0.05). The D68°C, mean D63°C, and D72°C for four M. avium subsp. paratuberculosis strains were 81.8, 9.8, and 4.2 s, respectively, yielding a mean z value of 6.9°C. Complete inactivation of 106 to 107 CFU of M. avium subsp. paratuberculosis/ml milk was not observed for any of the time-temperature combinations studied; 5.2- to 6.6-log10 reductions in numbers were achieved depending on the temperature and time. Nonlinear thermal inactivation kinetics were consistently observed for this bacterium. This study confirms that the optimized phage assay can be employed in place of conventional culture on HEYM to speed up the acquisition of results (48 h instead of a minimum of 6 weeks) for inactivation experiments involving M. avium subsp. paratuberculosis-spiked samples. PMID:20097817

Effect of single or combined chemical and natural antimicrobial interventions on Escherichia coli O157:H7, total microbiota and color of packaged spinach and lettuce.

PubMed

Poimenidou, Sofia V; Bikouli, Vasiliki C; Gardeli, Chryssavgi; Mitsi, Christina; Tarantilis, Petros A; Nychas, George-John; Skandamis, Panagiotis N

2016-03-02

Aqueous extract of Origanum vulgare (oregano), sodium hypochlorite (60 and 300 ppm of free chlorine), Citrox® (containing citric acid and phenolic compounds [bioflavonoids] as active ingredients), vinegar, lactic acid, and double combinations of Citrox, lactic acid and oregano were evaluated against Escherichia coli O157:H7 and total mesophilic microbiota on fresh-cut spinach and lettuce and for their impact on color of treated vegetables. Spinach and lettuce leaves were inoculated with E. coli O157:H7 to a level of 5-6 log CFU/g and immersed in washing solutions for 2 or 5 min at 20 °C, followed by rinsing with ice water (30s). Bacterial populations on vegetables were enumerated immediately after washing and after storage of the samples at 5 °C for 7 days under 20% CO2: 80% N2. No significant post-washing microbial reductions were achieved by chlorinated water, whereas after storage total microbiota was increased by 2.4 log CFU/g on lettuce. Vinegar wash was the most effective treatment causing E. coli O157:H7 reductions of 1.8-4.3 log CFU/g. During storage, pathogen was further decreased to below the detection limit level (<2 log CFU/g) and total microbiota exhibited the highest reductions compared to other treatments. Lactic acid reduced pathogen by 1.6-3.7 log CFU/g after washing; however levels of total microbiota increased by up to 2 log CFU/g on packaged lettuce during storage. Washing lettuce samples with oregano for 2 min resulted in 2.1 log CFU/g reduction of E. coli O157:H7. When Citrox was combined with oregano, 3.7-4.0 log CFU/g reduction was achieved on spinach and lettuce samples, with no significant effect on color parameters. Additionally, rinsing with ice water after decontamination treatments contributed to maintenance of color of the treated vegetables. In conclusion, the results indicated that vinegar, lactic acid or oregano aqueous extract alone or in combination, as alternative washing solutions to chlorine, may be effectively used to control E. coli O157:H7 and sustain acceptable appearance of fresh cut spinach and lettuce. Copyright © 2016 Elsevier B.V. All rights reserved.

Relating physicochemical and microbiological safety indicators during processing of linguiça, a Portuguese traditional dry-fermented sausage.

PubMed

Gonzales-Barron, U; Cadavez, V; Pereira, A P; Gomes, A; Araújo, J P; Saavedra, M J; Estevinho, L; Butler, F; Pires, P; Dias, T

2015-12-01

Linguiça is a Portuguese traditional fermented sausage whose microbiological quality and safety can be highly variable. In order to elucidate risk factors and the particularities of the manufacturing technology that explain the between-batch variability in total viable counts (TVC), Enterobacteriaceae, Staphylococcus aureus and Listeria monocytogenes in the product; microbiological and physicochemical characterisation of linguiça at five stages of production (i.e., raw pork meat, mixed with ingredients, macerated, smoked and ripened) was carried out. A total of six production batches were surveyed from two factories; one utilised curing salts and polyphosphate in their formulation (Factory II). The delayed fermentation in the nitrite-formulated sausages was partly responsible for the increase (p<0.01) in Enterobacteriaceae, S. aureus and L. monocytogenes from raw meat (3.21logCFU/g, 1.30logCFU/g and 22.2CFU/g, respectively) to the end of maceration (4.14logCFU/g, 2.10logCFU/g and 140CFU/g, respectively) while the better acidification process in the nitrite-free sausages (Factory I) led to lower counts of S. aureus (2.64logCFU/g) and L. monocytogenes (10CFU/g) in the finished products. In Factory II, although L. monocytogenes entered the chain at the point of mixing, it became steadily inactivated during smoking and ripening (<50CFU/g), despite the initially-delayed fermentation. Nitrite had a strong effect on reducing Enterobacteriaceae throughout smoking (r=-0.73) and ripening (r=-0.59), while it failed to control the growth of S. aureus. The main hurdle preventing the development of S. aureus in linguiça is the pH, and other factors contributing to its control are: longer ripening days (p=0.019), low S. aureus in raw meat (p=0.098), properly-washed casings (p=0.094), and less contamination during mixing (p=0.199). In the case of L. monocytogenes, at least three hurdles hinder its development in linguiça: low a w (p=0.004), low pH (p=0.040) and nitrite (p=0.060), and other factors contributing to its control are: longer ripening (p=0.072) and maceration (p=0.106) periods, lower a w at the end of smoking (p=0.076) and properly-washed casings (p=0.099). Results have shown that there is a need to standardise the productive process of linguiça, to optimise the initial acidification process, and to reinforce proper programmes of quality control of ingredients and good hygiene practices, so as to minimise the introduction of Enterobacteriaceae and pathogens from external sources. Copyright © 2015 Elsevier Ltd. All rights reserved.

Influence of mycorrhizal fungi on fate of E. coli O157:H7 and Salmonella in soil and internalization into Romaine lettuce plants.

PubMed

Nicholson, April M; Gurtler, Joshua B; Bailey, Rebecca B; Niemira, Brendan A; Douds, David D

2015-01-02

The objectives of this study were to determine the influence of a symbiotic arbuscular mycorrhizal (AM) fungus on persistence of Salmonella and enterohemorrhagic Escherichia coli O157:H7 (EHEC) within soil, and survival within Romaine lettuce. Romaine seedlings were grown with or without AM fungi. Soil surrounding plants was inoculated with ca. 8 log CFU/plant of either Salmonella enterica or E. coli EHEC composites. Samples (soil, root, and shoot) were analyzed on days 1, 8, 15 and 22 for Salmonella and EHEC by direct plating and selective enrichment. Twenty-four hours after inoculation, populations of Salmonella and EHEC, respectively, were 4.20 and 3.24 log CFU/root, 2.52 and 1.17 log CFU/shoot, and 5.46 and 5.17 log CFU/g soil. By selective enrichment, samples tested positive for Salmonella or EHEC at day 22 at rates of 94 and 68% (shoot), 97 and 56% (root), and 100 and 75% (soil), respectively, suggesting that Salmonella has a greater propensity for survival than EHEC. Salmonella populations in soil remained as high as 4.35 log CFU/g by day 22, while EHEC populations dropped to 1.12 log CFU/g in the same amount of time. Ninety-two percent of all Romaine leaves in our study were positive for internalized Salmonella from days 8 to 22 and remained as high as 1.26 log CFU/shoot on day 22 in AM fungi+Romaine plants. There were no differences (P>0.05) between the survival of either pathogen based on the presence or absence of mycorrhizal fungi. Results of this study suggest that AM fungi do not affect the internalization and/or survival of either S. enterica or E. coli O157:H7 in Romaine lettuce seedlings. Our results should provide Romaine lettuce farmers confidence that the presence and/or application of AM fungi to crop soil is not a contributing factor to the internalization and survival of Salmonella or E. coli O157:H7 within Romaine lettuce plants. Published by Elsevier B.V.

Performance of a completely automated system for monitoring CMV DNA in plasma.

PubMed

Mengelle, C; Sandres-Sauné, K; Mansuy, J-M; Haslé, C; Boineau, J; Izopet, J

2016-06-01

Completely automated systems for monitoring CMV-DNA in plasma samples are now available. Evaluate analytical and clinical performances of the VERIS™/MDx System CMV Assay(®). Analytical performance was assessed using quantified quality controls. Clinical performance was assessed by comparison with the COBAS(®) Ampliprep™/COBAS(®) Taqman CMV test using 169 plasma samples that had tested positive with the in-house technique in whole blood. The specificity of the VERIS™/MDx System CMV Assay(®) was 99% [CI 95%: 97.7-100]. Intra-assay reproducibilities were 0.03, 0.04, 0.05 and 0.04 log10IU/ml (means 2.78, 3.70, 4.64 and 5.60 log10IU/ml) for expected values of 2.70, 3.70, 4.70 and 5.70 log10IU/ml. The inter-assay reproducibilities were 0.12 and 0.08 (means 6.30 and 2.85 log10IU/ml) for expected values of 6.28 and 2.80 log10IU/ml. The lower limit of detection was 14.6IU/ml, and the assay was linear from 2.34 to 5.58 log10IU/ml. The results for the positive samples were concordant (r=0.71, p<0.0001; slope of Deming regression 0.79 [CI 95%: 0.56-1.57] and y-intercept 0.79 [CI 95%: 0.63-0.95]). The VERIS™/MDx System CMV Assay(®) detected 18 more positive samples than did the COBAS(®) Ampliprep™/COBAS(®) Taqman CMV test and the mean virus load were higher (0.41 log10IU/ml). Patient monitoring on 68 samples collected from 17 immunosuppressed patients showed similar trends between the two assays. As secondary question, virus loads detected by the VERIS™/MDx System CMV Assay(®) were compared to those of the in-house procedure on whole blood. The results were similar between the two assays (-0.09 log10IU/ml) as were the patient monitoring trends. The performances of the VERIS™/MDx System CMV Assay(®) facilitated its routine use in monitoring CMV-DNA loads in plasma samples. Copyright © 2016 Elsevier B.V. All rights reserved.

Biofilms associated with poultry processing equipment.

PubMed

Lindsay, D; Geornaras, I; von Holy, A

1996-01-01

Aerobic and Gram-negative bacteria were enumerated on non-metallic surfaces and stainless steel test pieces attached to equipment surfaces by swabbing and a mechanical dislodging procedure, respectively, in a South African grade B poultry processing plant. Changes in bacterial numbers were also monitored over time on metal test pieces. The highest bacterial counts were obtained from non-metallic surfaces such as rubber fingered pluckers and plastic defeathering curtains which exceeded the highest counts found on the metal surfaces by at least 1 log CFU cm-2. Gram-negative bacterial counts on all non-metallic surface types were at least 2 log CFU cm-2 lower than corresponding aerobic plate counts. On metal surfaces, the highest microbial numbers were obtained after 14 days exposure, with aerobic plate counts ranging from 3.57 log CFU cm-2 to 5.13 log CFU cm-2, and Gram-negative counts from 0.70 log CFU cm-2 to 3.31 log CFU cm-2. Scanning electron microscopy confirmed the presence of bacterial cells on non-metallic and metallic surfaces associated with poultry processing. Rubber 'fingers', plastic curtains, conveyor belt material and stainless steel test surfaces placed on the scald tank overflow and several chutes revealed extensive and often confluent bacterial biofilms. Extracellular polymeric substances, but few bacterial cells were visible on test pieces placed on evisceration equipment, spinchiller blades and the spinchiller outlet.

The effect of slaughter operations on the contamination of chicken carcasses with thermotolerant Campylobacter.

PubMed

Rosenquist, Hanne; Sommer, Helle M; Nielsen, Niels L; Christensen, Bjarke B

2006-04-25

To evaluate the effect of specific slaughter operations on the contamination of broiler carcasses with naturally occurring thermotolerant Campylobacter, experiments were carried out in two Danish commercial slaughter plants (Plant I and Plant II). Six broiler flocks determined Campylobacter positive prior to slaughter were investigated at four sampling locations within each slaughter plant. Quantification of thermotolerant Campylobacter in 30 neck skin samples per flock per sampling location showed that the evisceration operation in Plant I led to a significant increase in the Campylobacter concentration of 0.5 log(10) cfu/g in average, whereas no significant changes were observed during this operation in Plant II. Air chilling (Plant I) and water chilling (Plant II), both including a carcass wash prior to the chilling operation, caused similar, but significant reductions of 0.83 and 0.97 log(10) cfu/g, respectively. In packed frozen chickens (Plant II) an additional reduction of 1.38 log(10) cfu/g in average was obtained due to the freezing operation. In packed chilled chickens (Plant I), however, the number of thermotolerant Campylobacter per gram remained at the same level as after air chilling. Enumeration of thermotolerant Campylobacter in 30 intestinal samples per flock showed that in two of the six flocks examined the within flock colonization was very low (<3% and 27% positive samples). The remaining four flocks were colonized at percentages of 100 (three flocks) and 97 (one flock) and had intestinal mean counts ranging from 6.65 to 8.20 log(10) cfu/g. A correlation between Campylobacter concentrations in intestinal content and on chicken carcasses after the defeathering operation was documented. This finding indicates that a reduction in the Campylobacter concentration on chicken carcasses may also be obtained by interventions aimed at reducing the concentration of Campylobacter in the intestines of the living birds.

Behaviour of co-inoculated pathogenic and spoilage bacteria on poultry following several decontamination treatments.

PubMed

Alonso-Hernando, Alicia; Capita, Rosa; Alonso-Calleja, Carlos

2012-10-01

The potential of chemical decontaminants to cause harmful effects on human health is among the causes of the rejection of antimicrobial treatments for removing surface contamination from poultry carcasses in the European Union. This study was undertaken to determine whether decontaminants might give a competitive advantage to pathogenic bacteria on poultry and involve a potential risk to consumer. A total of 144 chicken legs were co-inoculated with similar concentrations of pathogenic bacteria (Listeria monocytogenes, Staphylococcus aureus, Salmonella enterica serotype Enteritidis or Escherichia coli) and spoilage bacteria (Brochothrix thermosphacta or Pseudomonas fluorescens). Samples were dipped for 15min in solutions (w/v) of trisodium phosphate (12%; TSP), acidified sodium chlorite (1200ppm; ASC), citric acid (2%; CA), peroxyacids (220ppm; PA) or chlorine dioxide (50ppm; CD), or were left untreated (control). Microbiological analyses were carried out on day 0 and every 24h until day 7 of storage (at 10±1°C). The modified Gompertz equation was used as the primary model to fit observed data. TSP, ASC and CA were effective in extending the lag phase (L, ranging from 1.47±1.34days to 4.06±1.16days) and in decreasing the concentration of bacteria during the stationary phase (D, ranging from 2.46±0.51 log(10) cfu/cm(2) to 8.64±0.53 log(10) cfu/cm(2)), relative to the control samples (L values ranging from 0.59±0.38days and 2.52±2.28days, and D values ranging from 6.32±0.89 log(10) cfu/cm(2) to 9.39±0.39 log(10) cfu/cm(2), respectively). Both on untreated and on most decontaminated samples the overgrowth of spoilage bacteria among the species tested was observed throughout storage, suggesting that spoilage would occur prior to any noteworthy increase in the levels of pathogenic microorganisms. However, L. monocytogenes counts similar to, or higher than, those for spoilage bacteria were observed on samples treated with TSP, ASC or CA, suggesting that these treatments might pose a danger to consumers. However, these results derive from laboratory-based experiments testing artificially-inoculated bacteria. Further investigations of the natural micro-biota of poultry carcasses would be appropriate in order to substantiate these findings. Copyright © 2012 Elsevier B.V. All rights reserved.

Sodium benzoate, potassium sorbate, and citric acid induce sublethal injury and enhance pulsed electric field inactivation of E. coli O157:H7 and nonpathogenic surrogate E. coli in strawberry juice

USDA-ARS?s Scientific Manuscript database

Current FDA regulations require that juice processors effect a 5 log CFU/ml reduction of a target pathogen prior to distributing products. Whereas thermal pasteurization reduces the sensory characteristics of juice by altering flavor components, pulsed electric field (PEF) treatment can be conducte...

Characterization of relative abundance of lactic acid bacteria species in French organic sourdough by cultural, qPCR and MiSeq high-throughput sequencing methods.

PubMed

Michel, Elisa; Monfort, Clarisse; Deffrasnes, Marion; Guezenec, Stéphane; Lhomme, Emilie; Barret, Matthieu; Sicard, Delphine; Dousset, Xavier; Onno, Bernard

2016-12-19

In order to contribute to the description of sourdough LAB composition, MiSeq sequencing and qPCR methods were performed in association with cultural methods. A panel of 16 French organic bakers and farmer-bakers were selected for this work. The lactic acid bacteria (LAB) diversity of their organic sourdoughs was investigated quantitatively and qualitatively combining (i) Lactobacillus sanfranciscensis-specific qPCR, (ii) global sequencing with MiSeq Illumina technology and (iii) molecular isolates identification. In addition, LAB and yeast enumeration, pH, Total Titratable Acidity, organic acids and bread specific volume were analyzed. Microbial and physico-chemical data were statistically treated by Principal Component Analysis (PCA) and Hierarchical Ascendant Classification (HAC). Total yeast counts were 6 log 10 to 7.6 log 10 CFU/g while LAB counts varied from 7.2 log 10 to 9.6 log 10 CFU/g. Values obtained by L. sanfranciscensis-specific qPCR were estimated between 7.2 and 10.3 log 10 CFU/g, except for one sample at 4.4 log 10 CFU/g. HAC and PCA clustered the sixteen sourdoughs into three classes described by their variables but without links to bakers' practices. L. sanfranciscensis was the dominant species in 13 of the 16 sourdoughs analyzed by Next Generation Sequencing (NGS), by the culture dependent method this species was dominant only in only 10 samples. Based on isolates identification, LAB diversity was higher for 7 sourdoughs with the recovery of L. curvatus, L. brevis, L. heilongjiangensis, L. xiangfangensis, L. koreensis, L. pontis, Weissella sp. and Pediococcus pentosaceus, as the most representative species. L. koreensis, L. heilongjiangensis and L. xiangfangensis were identified in traditional Asian food and here for the first time as dominant in organic sourdough. This study highlighted that L. sanfranciscensis was not the major species in 6/16 sourdough samples and that a relatively high LAB diversity can be observed in French organic sourdough. Copyright © 2016. Published by Elsevier B.V.

[Occurrence of bacteria in the mouth from genera of Micrococcus, Kocuria, Nesterenkonia, Kytococcus and Dermacoccus].

PubMed

Szczerba, Izabela; Krzemiński, Zbigniew

2002-01-01

The aim of the study was to assess the prevalence of different bacteria in the oral cavity. The bacteria were present in the oral cavities of 73 (48.7%) of 150 individuals. Nesterenkonia halobia, the most frequently isolated species, was found in 20 (27%) individuals, Micrococcus luteus in 16 (22%), Kocuria kristinae in 12 (16%), Kocuria varians in 10 (14%), Dermacoccus sedentarius in 9 (12%), Micrococcus lylae in 8 (11%), and Kytococcus nishinomiyaensis in 3 (4%). Mean counts of these microorganisms were relatively low and amounted in log10 CFU/ml saliva for M. luteus 1.87 +/- 0.52, for M. lylae 2.03 +/- 0.39, for N. halobia 2.14 +/- 0.56, for K. kristinae 2.20 +/- 0.69, for K. varians 2.19 +/- 0.67, for K. nishinomiyaensis 1.72 +/- 0.39, and for D. sedentarius 2.27 +/- 0.55. The factor limiting the population sizes of these microorganisms was most probably the antagonistic activity of the bacteria living in oral cavity.

Survival of Escherichia coli O157:H7 during manufacture and storage of white brined cheese.

PubMed

Osaili, Tareq M; Al-Nabulsi, Anas A; Olaimat, Amin N; Shaker, Reyad R; Taha, Mohammad; Holley, Richard A

2014-09-01

Escherichia coli O157:H7 is a major foodborne pathogen that causes severe disease in humans. Survival of E. coli O157:H7 during processing and storage of white brined cheese was investigated. Cheeses were prepared using pasteurized milk inoculated with a 4 strain E. coli O157:H7 cocktail (7 log(10) CFU/g) with or without yogurt starter culture (Lactobacillus delbrueckii ssp. bulgaricus and Streptococcus salivarius ssp. thermophilus) and stored in 10% or 15% NaCl brine at 10 and 21 ºC for 28 d. NaCl concentration, water activity (a(w)), pH, and numbers of E. coli O157:H7 and lactic acid bacteria (LAB) were determined in cheese and brine. E. coli O157:H7 was able to survive in cheese stored in both brines at 10 and 21 ºC regardless of the presence of starter LAB, although the latter significantly enhanced E. coli O157:H7 reduction in cheese or its brine at 10 ºC. E. coli O157:H7 numbers were reduced by 2.6 and 3.4 log(10) CFU/g in cheese stored in 10% and 15% NaCl brine, respectively, in the presence of starter LAB and by 1.4 and 2.3 log(10) CFU/g, respectively, in the absence of starter LAB at 10 ºC. The pathogen survived, but at lower numbers in the brines. The salt concentration of cheese stored in 10% brine remained about 5% during ripening, but in 15% brine, the NaCl level increased 1.6% to 8.1% (w/w) by 28 d. Values of pH and a(w) slightly decreased 1 d after exposure to brine and reached 5.5 to 6.6 and 0.88 to 0.94, respectively, in all treatments. © 2014 Institute of Food Technologists®

Reduction of Escherichia coli 0157:H7 in shredded iceberg lettuce by chlorination and gamma irradiation

NASA Astrophysics Data System (ADS)

Foley, D. M.; Dufour, A.; Rodriguez, L.; Caporaso, F.; Prakash, A.

2002-03-01

Lettuce was inoculated with a six-strain cocktail of acid-adapted Escherichia coli 0157:H7 at a level of 1×10 7 CFU/g. Following chlorination at 200 μg/ml, the lettuce was irradiated at 0.15, 0.38, or 0.55 kGy using a 60Co source. Survival of E. coli 0157:H7, aerobic mesophiles and yeast and molds were measured over a period of 10 days. For quality analysis, chlorinated lettuce was subjected to irradiation at 0.33 and 0.53 kGy and stored at 1.0°C, 4.0°C or 7.0°C. Changes in texture and color were determined by instrumental means and changes in flavor, odor, and visual quality were determined by sensory testing. Chlorination plus irradiation at 0.55 kGy produced a 5.4-log reduction in E. coli 0157:H7 levels. Chlorination alone reduced the E. coli 0157:H7 counts by 1-2 logs. Irradiation at 0.55 kGy was also effective in reducing standard plate counts and yeast and mold counts. Irradiation at this level did not cause softening of lettuce and sensory attributes were not adversely affected. In general, appearance and flavor were affected more by the length of storage than by temperature conditions. The 5+log reduction in E. coli counts and lack of adverse effects on sensory attributes indicate that low-dose irradiation can improve the safety and shelf-life of fresh-cut iceberg lettuce for retail sale or food service.

Efficacy of Gaseous Ozone Application during Vacuum Cooling against Escherichia coli O157:H7 on Spinach Leaves as Influenced by Bacterium Population Size.

PubMed

Yesil, Mustafa; Kasler, David R; Huang, En; Yousef, Ahmed E

2017-07-01

Foodborne disease outbreaks associated with the consumption of fresh produce pose a threat to public health, decrease consumer confidence in minimally processed foods, and negatively impact the sales of these commodities. The aim of the study was to determine the influence of population size of inoculated pathogen on its inactivation by gaseous ozone treatment during vacuum cooling. Spinach leaves were spot inoculated with Escherichia coli O157:H7 at approximate initial populations of 10 8 , 10 7 , and 10 5 CFU/g. Inoculated leaves were vacuum cooled (28.5 inHg; 4°C) in a custom-made vessel and then were subjected to a gaseous ozone treatment under the following conditions: 1.5 g of ozone per kg of gas mixture, vessel pressure at 10 lb/in 2 gauge, 94 to 98% relative humidity, and 30 min of holding time at 9°C. Treatment of the leaves, having the aforementioned inocula, decreased E. coli populations by 0.2, 2.1, and 2.8 log CFU/g, respectively, compared with the inoculated untreated controls. Additionally, spinach leaves were inoculated at 1.4 × 10 3 CFU/g, which approximates natural contamination level, and the small populations remaining after ozone treatment were quantified using the most-probable-number (MPN) method. Vacuum and ozone sequential treatment decreased this E. coli O157:H7 population to <3 MPN/g (i.e., greater than 3-log reduction). Resulting log reductions were greater (P < 0.05) at the lower rather than the higher inoculum levels. In conclusion, treatment of spinach leaves with gaseous ozone is effective against pathogen loads comparable to those found in naturally contaminated fresh produce, but efficacy decreases as inoculum level increases.

A novel method to prepare concentrated conidial biomass formulation of Trichoderma harzianum for seed application.

PubMed

Singh, P C; Nautiyal, C S

2012-12-01

To prepare concentrated formulation of Trichoderma harzianum MTCC-3841 (NBRI-1055) with high colony forming units (CFU), long shelf life and efficient in root colonization by a simple scrapping method. NBRI-1055 spores scrapped from potato dextrose agar plates were used to prepare a concentrated formulation after optimizing carrier material, moisture content and spore harvest time. The process provides an advantage of maintaining optimum moisture level by the addition of water rather than dehydration. The formulation had an initial 11-12 log(10) CFU g(-1). Its concentrated form reduces its application amount by 100 times (10 g 100 kg(-1) seed) and provides 3-4 log(10) CFU seed(-1). Shelf life of the product was experimentally determined at 30 and 40 °C and predicted at other temperatures following Arrhenius equation. The concentrated formulation as compared to similar products provides an extra advantage of smaller packaging for storage and transportation, cutting down product cost. Seed application of the formulation recorded significant increase in plant growth promotion. Stable and effective formulation of Trichoderma harzianum NBRI-1055 was obtained by a simple scrapping method. A new method for the production of concentrated, stable, effective and cost efficient formulation of T. harzianum has been validated for seed application. © 2012 The Society for Applied Microbiology.

Tracheal tube biofilm removal through a novel closed-suctioning system: an experimental study.

PubMed

Aguilera Xiol, E; Li Bassi, G; Wyncoll, D; Ntoumenopoulos, G; Fernandez-Barat, L; Marti, J D; Comaru, T; De Rosa, F; Rigol, M; Rinaudo, M; Ferrer, M; Torres, A

2015-11-01

Tracheal tube biofilm develops during mechanical ventilation. We compared a novel closed-suctioning system vs standard closed-suctioning system in the prevention of tracheal tube biofilm. Eighteen pigs, on mechanical ventilation for 76 h, with P. aeruginosa pneumonia were randomized to be tracheally suctioned via the KIMVENT* closed-suctioning system (control group) or a novel closed-suctioning system (treatment group), designed to remove tracheal tube biofilm through saline jets and an inflatable balloon. Upon autopsy, two tracheal tube hemi-sections were dissected for confocal and scanning electron microscopy. Biofilm area, maximal and minimal thickness were computed. Biofilm stage was assessed. Sixteen animals were included in the final analysis. In the treatment and control group, the mean (sd) pulmonary burden was 3.34 (1.28) and 4.17 (1.09) log cfu gr(-1), respectively (P=0.18). Tracheal tube P. aeruginosa colonization was 5.6 (4.9-6.3) and 6.2 (5.6-6.9) cfu ml(-1) (median and interquartile range) in the treatment and control group, respectively (P=0.23). In the treatment group, median biofilm area was 3.65 (3.22-4.21) log10 μm2 compared with 4.49 (4.27-4.52) log10 μm2 in the control group (P=0.031). In the treatment and control groups, the maximal biofilm thickness was 48.3 (26.7-71.2) µm (median and interquartile range) and 88.8 (43.8-125.7) µm, respectively. The minimal thickness in the treatment and control group was 0.6 (0-4.0) µm and 23.7 (5.3-27.8) µm (P=0.040) (P=0.017). Earlier stages of biofilm development were found in the treatment group (P<0.001). The novel CSS reduces biofilm accumulation within the tracheal tube. A clinical trial is required to confirm these findings and the impact on major outcomes. © The Author 2015. Published by Oxford University Press on behalf of the British Journal of Anaesthesia. All rights reserved. For Permissions, please email: journals.permissions@oup.com.

Real-Time PCR Detection of Paenibacillus spp. in Raw Milk To Predict Shelf Life Performance of Pasteurized Fluid Milk Products

PubMed Central

Ranieri, Matthew L.; Ivy, Reid A.; Mitchell, W. Robert; Call, Emma; Masiello, Stephanie N.; Wiedmann, Martin

2012-01-01

Psychrotolerant sporeformers, specifically Paenibacillus spp., are important spoilage bacteria for pasteurized, refrigerated foods such as fluid milk. While Paenibacillus spp. have been isolated from farm environments, raw milk, processing plant environments, and pasteurized fluid milk, no information on the number of Paenibacillus spp. that need to be present in raw milk to cause pasteurized milk spoilage was available. A real-time PCR assay targeting the 16S rRNA gene was designed to detect Paenibacillus spp. in fluid milk and to discriminate between Paenibacillus and other closely related spore-forming bacteria. Specificity was confirmed using 16 Paenibacillus and 17 Bacillus isolates. All 16 Paenibacillus isolates were detected with a mean cycle threshold (CT) of 19.14 ± 0.54. While 14/17 Bacillus isolates showed no signal (CT > 40), 3 Bacillus isolates showed very weak positive signals (CT = 38.66 ± 0.65). The assay provided a detection limit of approximately 3.25 × 101 CFU/ml using total genomic DNA extracted from raw milk samples inoculated with Paenibacillus. Application of the TaqMan PCR to colony lysates obtained from heat-treated and enriched raw milk provided fast and accurate detection of Paenibacillus. Heat-treated milk samples where Paenibacillus (≥1 CFU/ml) was detected by this colony TaqMan PCR showed high bacterial counts (>4.30 log CFU/ml) after refrigerated storage (6°C) for 21 days. We thus developed a tool for rapid detection of Paenibacillus that has the potential to identify raw milk with microbial spoilage potential as a pasteurized product. PMID:22685148