Comparison of viable plate count, turbidity measurement and real-time PCR for quantification of Porphyromonas gingivalis.

PubMed

Clais, S; Boulet, G; Van Kerckhoven, M; Lanckacker, E; Delputte, P; Maes, L; Cos, P

2015-01-01

The viable plate count (VPC) is considered as the reference method for bacterial enumeration in periodontal microbiology but shows some important limitations for anaerobic bacteria. As anaerobes such as Porphyromonas gingivalis are difficult to culture, VPC becomes time-consuming and less sensitive. Hence, efficient normalization of experimental data to bacterial cell count requires alternative rapid and reliable quantification methods. This study compared the performance of VPC with that of turbidity measurement and real-time PCR (qPCR) in an experimental context using highly concentrated bacterial suspensions. Our TaqMan-based qPCR assay for P. gingivalis 16S rRNA proved to be sensitive and specific. Turbidity measurements offer a fast method to assess P. gingivalis growth, but suffer from high variability and a limited dynamic range. VPC was very time-consuming and less repeatable than qPCR. Our study concludes that qPCR provides the most rapid and precise approach for P. gingivalis quantification. Although our data were gathered in a specific research context, we believe that our conclusions on the inferior performance of VPC and turbidity measurements in comparison to qPCR can be extended to other research and clinical settings and even to other difficult-to-culture micro-organisms. Various clinical and research settings require fast and reliable quantification of bacterial suspensions. The viable plate count method (VPC) is generally seen as 'the gold standard' for bacterial enumeration. However, VPC-based quantification of anaerobes such as Porphyromonas gingivalis is time-consuming due to their stringent growth requirements and shows poor repeatability. Comparison of VPC, turbidity measurement and TaqMan-based qPCR demonstrated that qPCR possesses important advantages regarding speed, accuracy and repeatability. © 2014 The Society for Applied Microbiology.

APPARENT BIAS IN RIVER WATER INOCULUM FOLLOWING CENTRIFUGATION

EPA Science Inventory

We collected four measures of viable bacterial concentration (heterotrophic plate count, total coliform, fecal coliform, and Escherichia coli) and three measures of well color development in Biolog GN2 microtiter plates from water samples that were collected on two or three separ...

In vitro effect of Reiki treatment on bacterial cultures: Role of experimental context and practitioner well-being.

PubMed

Rubik, Beverly; Brooks, Audrey J; Schwartz, Gary E

2006-01-01

To measure effects of Reiki treatments on growth of heat-shocked bacteria, and to determine the influence of healing context and practitioner well-being. Overnight cultures of Escherichia coli K12 in fresh medium were used. Culture samples were paired with controls to minimize any ordering effects. Samples were heat-shocked prior to Reiki treatment, which was performed by Reiki practitioners for up to 15 minutes, with untreated controls. Plate-count assay using an automated colony counter determined the number of viable bacteria. Fourteen Reiki practitioners each completed 3 runs (n = 42 runs) without healing context, and another 2 runs (n = 28 runs) in which they first treated a pain patient for 30 minutes (healing context). Well-being questionnaires were administered to practitioners pre-post all sessions. No overall difference was found between the Reiki and control plates in the nonhealing context. In the healing context, the Reiki treated cultures overall exhibited significantly more bacteria than controls (p < 0.05). Practitioner social (p < 0.013) and emotional well-being (p < 0.021) correlated with Reiki treatment outcome on bacterial cultures in the nonhealing context. Practitioner social (p < 0.031), physical (p < 0.030), and emotional (p < 0.026) well-being correlated with Reiki treatment outcome on the bacterial cultures in the healing context. For practitioners starting with diminished well-being, control counts were likely to be higher than Reiki-treated bacterial counts. For practitioners starting with a higher level of well-being, Reiki counts were likely to be higher than control counts. Reiki improved growth of heat-shocked bacterial cultures in a healing context. The initial level of well-being of the Reiki practitioners correlates with the outcome of Reiki on bacterial culture growth and is key to the results obtained.

Steam versus hot-water scalding in reducing bacterial loads on the skin of commercially processed poultry.

PubMed

Patrick, T E; Goodwin, T L; Collins, J A; Wyche, R C; Love, B E

1972-04-01

A comparison of two types of scalders was conducted to determine their effectiveness in reducing bacterial contamination of poultry carcasses. A conventional hot-water scalder and a prototype model of a steam scalder were tested under commercial conditions. Total plate counts from steam-scalded birds were significantly lower than the counts of water-scalded birds immediately after scalding and again after picking. No differences in the two methods could be found after chilling. Coliform counts from steam-scalded birds were significantly lower than the counts from water-scalded birds immediately after scalding. No significant differences in coliform counts were detected when the two scald methods were compared after defeathering and chilling.

Steam Versus Hot-Water Scalding in Reducing Bacterial Loads on the Skin of Commercially Processed Poultry

PubMed Central

Patrick, Thomas E.; Goodwin, T. L.; Collins, J. A.; Wyche, R. C.; Love, B. E.

1972-01-01

A comparison of two types of scalders was conducted to determine their effectiveness in reducing bacterial contamination of poultry carcasses. A conventional hot-water scalder and a prototype model of a steam scalder were tested under commercial conditions. Total plate counts from steam-scalded birds were significantly lower than the counts of water-scalded birds immediately after scalding and again after picking. No differences in the two methods could be found after chilling. Coliform counts from steam-scalded birds were significantly lower than the counts from water-scalded birds immediately after scalding. No significant differences in coliform counts were detected when the two scald methods were compared after defeathering and chilling. PMID:4553146

Responses of Escherichia coli, Listeria monocytogenes, and Staphylococcus aureus to Simulated Food Processing Treatments, Determined Using Fluorescence-Activated Cell Sorting and Plate Counting▿

PubMed Central

Kennedy, Deirdre; Cronin, Ultan P.; Wilkinson, Martin G.

2011-01-01

Three common food pathogenic microorganisms were exposed to treatments simulating those used in food processing. Treated cell suspensions were then analyzed for reduction in growth by plate counting. Flow cytometry (FCM) and fluorescence-activated cell sorting (FACS) were carried out on treated cells stained for membrane integrity (Syto 9/propidium iodide) or the presence of membrane potential [DiOC2(3)]. For each microbial species, representative cells from various subpopulations detected by FCM were sorted onto selective and nonselective agar and evaluated for growth and recovery rates. In general, treatments giving rise to the highest reductions in counts also had the greatest effects on cell membrane integrity and membrane potential. Overall, treatments that impacted cell membrane permeability did not necessarily have a comparable effect on membrane potential. In addition, some bacterial species with extensively damaged membranes, as detected by FCM, appeared to be able to replicate and grow after sorting. Growth of sorted cells from various subpopulations was not always reflected in plate counts, and in some cases the staining protocol may have rendered cells unculturable. Optimized FCM protocols generated a greater insight into the extent of the heterogeneous bacterial population responses to food control measures than did plate counts. This study underlined the requirement to use FACS to relate various cytometric profiles generated by various staining protocols with the ability of cells to grow on microbial agar plates. Such information is a prerequisite for more-widespread adoption of FCM as a routine microbiological analytical technique. PMID:21602370

Bacterial Cleanability of Various Types of Eating Surfaces.

ERIC Educational Resources Information Center

Ridenour, Gerald M.; Armbruster, E. H.

1953-01-01

Presents a study of the capability of commercial dishwashers to remove bacteria from various kinds of service plates. Gives an account of preliminary research on the bacterial cleanability of eating surfaces of different materials by two radiological procedures--(1) radiological count, and (2) autoradiographic measurement. Among the factors…

Flow cytometric bacterial cell counts challenge conventional heterotrophic plate counts for routine microbiological drinking water monitoring.

PubMed

Van Nevel, S; Koetzsch, S; Proctor, C R; Besmer, M D; Prest, E I; Vrouwenvelder, J S; Knezev, A; Boon, N; Hammes, F

2017-04-15

Drinking water utilities and researchers continue to rely on the century-old heterotrophic plate counts (HPC) method for routine assessment of general microbiological water quality. Bacterial cell counting with flow cytometry (FCM) is one of a number of alternative methods that challenge this status quo and provide an opportunity for improved water quality monitoring. After more than a decade of application in drinking water research, FCM methodology is optimised and established for routine application, supported by a considerable amount of data from multiple full-scale studies. Bacterial cell concentrations obtained by FCM enable quantification of the entire bacterial community instead of the minute fraction of cultivable bacteria detected with HPC (typically < 1% of all bacteria). FCM measurements are reproducible with relative standard deviations below 3% and can be available within 15 min of samples arriving in the laboratory. High throughput sample processing and complete automation are feasible and FCM analysis is arguably less expensive than HPC when measuring more than 15 water samples per day, depending on the laboratory and selected staining procedure(s). Moreover, many studies have shown FCM total (TCC) and intact (ICC) cell concentrations to be reliable and robust process variables, responsive to changes in the bacterial abundance and relevant for characterising and monitoring drinking water treatment and distribution systems. The purpose of this critical review is to initiate a constructive discussion on whether FCM could replace HPC in routine water quality monitoring. We argue that FCM provides a faster, more descriptive and more representative quantification of bacterial abundance in drinking water. Copyright © 2017 Elsevier Ltd. All rights reserved.

Transit time affects the community stability of Lactobacillus and Bifidobacterium species in an in vitro model of human colonic microbiotia.

PubMed

Rodes, Laetitia; Paul, Arghya; Coussa-Charley, Michael; Al-Salami, Hani; Tomaro-Duchesneau, Catherine; Fakhoury, Marc; Prakash, Satya

2011-12-01

Retention time, which is analogous to transit time, is an index for bacterial stability in the intestine. Its consideration is of particular importance to optimize the delivery of probiotic bacteria in order to improve treatment efficacy. This study aims to investigate the effect of retention time on Lactobacilli and Bifidobacteria stability using an established in vitro human colon model. Three retention times were used: 72, 96, and 144 h. The effect of retention time on cell viability of different bacterial populations was analyzed with bacterial plate counts and PCR. The proportions of intestinal Bifidobacteria, Lactobacilli, Enterococci, Staphylococci and Clostridia populations, analyzed by plate counts, were found to be the same as that in human colonic microbiota. Retention time in the human colon affected the stability of Lactobacilli and Bifidobacteria communities, with maximum stability observed at 144 h. Therefore, retention time is an important parameter that influences bacterial stability in the colonic microbiota. Future clinical studies on probiotic bacteria formulations should take into consideration gastrointestinal transit parameters to improve treatment efficacy.

Comparison of solid-phase cytometry and the plate count method for the evaluation of the survival of bacteria in pharmaceutical oils.

PubMed

De Prijck, K; Peeters, E; Nelis, H J

2008-12-01

To compare the survival of four bacterial strains (Escherichia coli, Proteus mirabilis, Staphylococcus aureus, Pseudomonas aeruginosa) in pharmaceutical oils, including jojoba oil/tea tree oil, carbol oil, jojoba oil and sesame oil. Oils were spiked with the test bacteria in a concentration of 10(4) CFU ml(-1). Bacteria were extracted from oils with phosphate-buffered saline containing 0.5% Tween 20. Aliquots of the pooled water layers were analysed by solid-phase cytometry and plate counting. Plate counts dropped to zero for all test strains exposed for 24 h to three of the four oils. In contrast, significant numbers of viable cells were still detected by SPC, except in the jojoba oil/tea tree oil mixture and partly in sesame oil. Exposure of bacteria for 24 h to the two oils containing an antimicrobial led to a loss of their culturability but not necessarily of their viability. The antibacterial activity of the jojoba oil/tea tree oil mixture supersedes that of carbol oil. These in vitro data suggest that the jojoba oil/tea tree oil mixture more than carbol oil inhibits bacterial proliferation when used for intermittent self-catherization.

The use of multiple indices of physiological activity to access viability in chlorine disinfected Escherichia coli O157:H7

NASA Technical Reports Server (NTRS)

Lisle, J. T.; Pyle, B. H.; McFeters, G. A.

1999-01-01

A suite of fluorescent intracellular stains and probes was used, in conjunction with viable plate counts, to assess the effect of chlorine disinfection on membrane potential (rhodamine 123; Rh123 and bis-(1,3-dibutylbarbituric acid) trimethine oxonol; DiBAC4(3)), membrane integrity (LIVE/DEAD BacLight kit), respiratory activity (5-cyano-2,3-ditolyl tetrazolium chloride; CTC) and substrate responsiveness (direct viable counts; DVC) in the commensal pathogen Escherichia coli O157:H7. After a 5 min exposure to the disinfectant, physiological indices were affected in the following order: viable plate counts > substrate responsiveness > membrane potential > respiratory activity > membrane integrity. In situ assessment of physiological activity by examining multiple targets, as demonstrated in this study, permits a more comprehensive determination of the site and extent of injury in bacterial cells following sublethal disinfection with chlorine. This approach to assessing altered bacterial physiology has application in various fields where detection of stressed bacteria is of interest.

Small-Scale Vertical Distribution of Bacterial Biomass and Diversity in Biological Soil Crusts from Arid Lands in the Colorado Plateau

USGS Publications Warehouse

Garcia-Pichel, F.; Johnson, S.L.; Youngkin, D.; Belnap, J.

2003-01-01

We characterized, at millimeter resolution, bacterial biomass, diversity, and vertical stratification of biological soil crusts in arid lands from the Colorado Plateau. Microscopic counts, extractable DNA, and plate counts of viable aerobic copiotrophs (VAC) revealed that the top centimeter of crusted soils contained atypically large bacterial populations, tenfold larger than those in uncrusted, deeper soils. The plate counts were not always consistent with more direct estimates of microbial biomass. Bacterial populations peaked at the immediate subsurface (1-2 mm) in light-appearing, young crusts, and at the surface (0-1 mm) in well-developed, dark crusts, which corresponds to the location of cyanobacterial populations. Bacterial abundance decreased with depth below these horizons. Spatially resolved DGGE fingerprints of Bacterial 16S rRNA genes demonstrated the presence of highly diverse natural communities, but we could detect neither trends with depth in bacterial richness or diversity, nor a difference in diversity indices between crust types. Fingerprints, however, revealed the presence of marked stratification in the structure of the microbial communities, probably a result of vertical gradients in physicochemical parameters. Sequencing and phylogenetic analyses indicated that most of the naturally occurring bacteria are novel types, with low sequence similarity (83-93%) to those available in public databases. DGGE analyses of the VAC populations indicated communities of lower diversity, with most types having sequences more than 94% similar to those in public databases. Our study indicates that soil crusts represent small-scale mantles of fertility in arid ecosystems, harboring vertically structured, little-known bacterial populations that are not well represented by standard cultivation methods.

An evidential example of airborne bacteria in a crowded, underground public concourse in Tokyo

NASA Astrophysics Data System (ADS)

Seino, Kaoruko; Takano, Takehito; Nakamura, Keiko; Watanabe, Masafumi

2005-01-01

We examined airborne bacteria in an underground concourse in Tokyo and investigated conditions that influenced bacterial counts. Airborne bacteria were collected by using an impactor sampler. Colonies on plate count agar (PCA) and Columbia colistin-nalidixic acid agar with 5% sheep blood (CNA agar) were enumerated. The range, geometric mean, and 95% CI of the bacterial counts (CFU m-3) on PCA and CNA agar were 150-1380, 456, 382-550 and 50-990, 237, 182-309, respectively. Bacterial counts on PCA significantly correlated with number of the pedestrians (r=0.89), relative humidity (r=0.70) and airborne dust (PM5.0) (r=0.73). Results of a multiple regression indicated independent positive association between the number of pedestrians and bacterial counts on PCA (p<0.01) after excluding the influence of relative humidity and airborne dust. Similar results were obtained with the statistical analysis for the counts of bacteria on CNA agar. Gram-positive cocci were dominant on PCA and CNA agar. Staphylococcus epidermidis and Micrococcus spp. were dominant among the 11 genera and 19 species identified in the present study. Considering the pattern of identified species and the significant independent association between number of pedestrians and bacterial counts, airborne bacteria in a crowded underground concourse were mostly originated from the pedestrians who were walking in the underground concourse. This study gave an evidential example of bacterial conditions in the air of an underground crowded public space in Tokyo.

Nonrecovery of varying proportions of viable bacteria during spread plating governed by the extent of spreader usage and proposal for an alternate spotting-spreading approach to maximize the CFU.

PubMed

Thomas, P; Sekhar, A C; Mujawar, M M

2012-08-01

To elucidate the cause of high variations and inconsistencies in bacterial CFU observed within and between different experiments while assessing viable bacterial counts through spread plating (SP). Following the inconsistent results, CFU estimations were undertaken through conventional SP using the spreader, or a modified approach that did not use spreader employing four organisms. The latter approach involving spotting-and-tilt-spreading of inoculum on agar surface [spotting spreading (SS)] yielded higher CFU by 11-120% over the weighted average depending on the organism and diluent. The adverse effect owing to the spreader was the most obvious in Escherichia coli followed by Staphylococcus epidermidis, Enterobacter cloacae and Bacillus pumilus. Plate attributes that determined the surface moisture levels of agar medium and the spreading practice adopted by the personnel formed two other major influencing factors. Plating for shorter periods (<60 s) using fresh 15/20 ml plates caused loss of 3-12% CFU owing to inoculum adhesion to spreader irrespective of glass or polypropylene make. On the other hand, prolonging the plating brought down the CFU significantly. Spreader movement on agar surface subsequent to the exhaustion of free moisture, which was marked by the experiencing of some friction to smooth spreader movement, was detrimental to vegetative cells, while Bacillus spores were less affected. The study brings out that the way SP is carried out exerts significant effects on CFU influenced by plate conditions. Prolonged use of spreader on dry agar surface could be highly detrimental to bacterial cells. A mild use of spreader accounting for spreader-adhering inoculum or the practice of SS not involving the spreader is recommended. This study unravels the effects owing to the spreader on bacterial cells and the CFU and recommends an alternate approach of SS to minimize CFU inconsistencies and to maximize the viable bacterial counts. © 2012 The Authors Journal of Applied Microbiology © 2012 The Society for Applied Microbiology.

Evaluation of heterotrophic plate and chromogenic agar colony counting in water quality laboratories.

PubMed

Hallas, Gary; Monis, Paul

2015-01-01

The enumeration of bacteria using plate-based counts is a core technique used by food and water microbiology testing laboratories. However, manual counting of bacterial colonies is both time and labour intensive, can vary between operators and also requires manual entry of results into laboratory information management systems, which can be a source of data entry error. An alternative is to use automated digital colony counters, but there is a lack of peer-reviewed validation data to allow incorporation into standards. We compared the performance of digital counting technology (ProtoCOL3) against manual counting using criteria defined in internationally recognized standard methods. Digital colony counting provided a robust, standardized system suitable for adoption in a commercial testing environment. The digital technology has several advantages:•Improved measurement of uncertainty by using a standard and consistent counting methodology with less operator error.•Efficiency for labour and time (reduced cost).•Elimination of manual entry of data onto LIMS.•Faster result reporting to customers.

The effect of different precooling rates and cold storage on milk microbiological quality and composition.

PubMed

Paludetti, Lizandra F; Kelly, Alan L; O'Brien, Bernadette; Jordan, Kieran; Gleeson, David

2018-03-01

The objective of this study was to measure the effect of different milk cooling rates, before entering the bulk tank, on the microbiological load and composition of the milk, as well as on energy usage. Three milk precooling treatments were applied before milk entered 3 identical bulk milk tanks: no plate cooler (NP), single-stage plate cooler (SP), and double-stage plate cooler (DP). These precooling treatments cooled the milk to 32.0 ± 1.4°C, 17.0 ± 2.8°C, and 6.0 ± 1.1°C, respectively. Milk was added to the bulk tank twice daily for 72 h, and the tank refrigeration temperature was set at 3°C. The blend temperature within each bulk tank was reduced after each milking event as the volume of milk at 3°C increased simultaneously. The bacterial counts of the milk volumes precooled at different rates did not differ significantly at 0 h of storage or at 24-h intervals thereafter. After 72 h of storage, the total bacterial count of the NP milk was 3.90 ± 0.09 log 10 cfu/mL, whereas that of the precooled milk volumes were 3.77 ± 0.09 (SP) and 3.71 ± 0.09 (DP) log 10 cfu/mL. The constant storage temperature (3°C) over 72 h helped to reduce bacterial growth rates in milk; consequently, milk composition was not affected and minimal, if any, proteolysis occurred. The DP treatment had the highest energy consumption (17.6 ± 0.5 Wh/L), followed by the NP (16.8 ± 2.7 Wh/L) and SP (10.6 ± 1.3 Wh/L) treatments. This study suggests that bacterial count and composition of milk are minimally affected when milk is stored at 3°C for 72 h, regardless of whether the milk is precooled; however, milk entering the tank should have good initial microbiological quality. Considering the numerical differences between bacterial counts, however, the use of the SP or DP precooling systems is recommended to maintain low levels of bacterial counts and reduce energy consumption. The Authors. Published by FASS Inc. and Elsevier Inc. on behalf of the American Dairy Science Association®. This is an open access article under the CC BY-NC-ND license (http://creativecommons.org/licenses/by-nc-nd/3.0/).

Effectiveness of a steam cleaning unit for disinfection in a veterinary hospital.

PubMed

Wood, Cheryl L; Tanner, Benjamin D; Higgins, Laura A; Dennis, Jeffrey S; Luempert, Louis G

2014-12-01

To evaluate whether the application of steam to a variety of surface types in a veterinary hospital would effectively reduce the number of bacteria. 5 surface types. Steam was applied as a surface treatment for disinfection to 18 test sites of 5 surface types in a veterinary hospital. A pretreatment sample was obtained by collection of a swab specimen from the left side of each defined test surface. Steam disinfection was performed on the right side of each test surface, and a posttreatment sample was then collected in the same manner from the treated (right) side of each test surface. Total bacteria for pretreatment and posttreatment samples were quantified by heterotrophic plate counts and for Staphylococcus aureus, Pseudomonas spp, and total coliforms by counts on selective media. Significant reductions were observed in heterotrophic plate counts after steam application to dog runs and dog kennel floors. A significant reduction in counts of Pseudomonas spp was observed after steam application to tub sinks. Bacterial counts were reduced, but not significantly, on most other test surfaces that had adequate pretreatment counts for quantification. Development of health-care-associated infections is of increasing concern in human and veterinary medicine. The application of steam significantly reduced bacterial numbers on a variety of surfaces within a veterinary facility. Steam disinfection may prove to be an alternative or adjunct to chemical disinfection within veterinary practices.

Escherichia coli viability determination using dynamic light scattering: a comparison with standard methods.

PubMed

Loske, Achim M; Tello, Elba M; Vargas, Susana; Rodriguez, Rogelio

2014-08-01

To determine the concentration of bacteria in a sample is important in the food industry, medicine and biotechnology. A disadvantage of the plate-counting method is that a microorganism colony could arise from one cell or from many cells. The other standard methodology, known as optical density determination, is based on the turbidity of a suspension and registers all bacteria, dead and alive. In this article, dynamic light scattering is proposed as a fast and reliable method to determine bacterial viability and, consequently, time evolution. Escherichia coli was selected because this microorganism is well known and easy to handle. A correlation between the data from these three techniques was obtained. We were able to calculate the growth rate, usually determined by plate counting or optical density measurement, using dynamic light scattering and to predict bacterial behavior. An analytical relationship between the colony forming units and the light scattered intensity was also deduced.

Fortifying fresh human milk with commercial powdered human milk fortifiers does not affect bacterial growth during 6 hours at room temperature.

PubMed

Telang, Sucheta; Berseth, Carol Lynn; Ferguson, Paul W; Kinder, Julie M; DeRoin, Mark; Petschow, Bryon W

2005-10-01

To evaluate the growth of resident aerobic mesophilic flora and added Enterobacter sakazakii in fresh, unfortified human milk; fresh human milk fortified with two commercial powdered fortifiers differing in iron content; and infant formula prepared from powder. Eight mothers provided preterm breast milk samples. Breast milk samples were divided into three aliquots: unfortified, fortified with fortifier containing 1.44 mg iron/14 kcal, and fortified with fortifier containing 0.4 mg iron/14 kcal. Aliquots of formula were prepared. Breast milk and formula aliquots were divided into two test samples. Half were inoculated with low amounts of E sakazakii; half were not. All test samples were maintained at room temperature (22 degrees C), serially diluted, and plated onto agars after 0, 2, 4, and 6 hours. Plates were incubated at 35 degrees C and enumerated. Data were analyzed using repeated measures analysis of variance. P<.05 was considered significant. There were no differences in colony counts of aerobic bacteria among uninoculated or among inoculated human milk samples at any time; counts did not increase significantly over 6 hours. There were no differences in colony counts of E sakazakii among inoculated human milk samples at any time; counts did not increase significantly over 6 hours. Aerobic bacteria and E sakazakii colony counts from infant formula did not increase significantly over 6 hours. During 6 hours at 22 degrees C, fresh human milk and formula had negligible bacterial growth; fortifying human milk with powdered fortifiers did not affect bacterial growth.

A rapid detection method using flow cytometry to monitor the risk of Legionella in bath water.

PubMed

Taguri, Toshitsugu; Oda, Yasunori; Sugiyama, Kanji; Nishikawa, Toru; Endo, Takuro; Izumiyama, Shinji; Yamazaki, Masayuki; Kura, Fumiaki

2011-07-01

Legionella species are the causative agents of human legionellosis, and bathing facilities have been identified as the sources of infection in several outbreaks in Japan. Researchers in Japan have recently reported evidence of significant associations between bacterial counts and the occurrence of Legionella in bathing facilities and in a hot tub model. A convenient and quantitative bacterial enumeration method is therefore required as an indicator of Legionella contamination or disinfection to replace existing methods such as time-consuming Legionella culture and expensive Legionella-DNA amplification. In this study, we developed a rapid detection method (RDM) to monitor the risk of Legionella using an automated microbial analyzing device based on flow cytometry techniques to measure the total number of bacteria in water samples within two minutes, by detecting typical patterns of scattered light and fluorescence. We first compared the results of our RDM with plate counting results for five filtered hot spring water samples spiked with three species of bacteria, including Legionella. Inactivation of these samples by chlorine was also assessed by the RDM, a live/dead bacterial fluorescence assay and plate counting. Using the RDM, the lower limit of quantitative bacterial counts in the spiked samples was determined as 3.0×10(3)(3.48log)counts mL(-1). We then used a laboratory model of a hot tub and found that the RDM could monitor the growth curve of naturally occurring heterotrophic bacteria with 1 and 2 days' delayed growth of amoeba and Legionella, respectively, and could also determine the killing curve of these bacteria by chlorination. Finally, samples with ≥3.48 or <3.48log total bacterial counts mL(-1) were tested using the RDM from 149 different hot tubs, and were found to be significantly associated with the positive or negative detection of Legionella with 95% sensitivity and 84% specificity. These findings indicated that the RDM can be used for Legionella control at bathing facilities, especially those where the effectiveness of chlorine is reduced by the presence of Fe(2+), Mn(2+), NH(4)(+), skin debris, and/or biofilms in the water. Copyright © 2011 Elsevier B.V. All rights reserved.

Automated food microbiology: potential for the hydrophobic grid-membrane filter.

PubMed Central

Sharpe, A N; Diotte, M P; Dudas, I; Michaud, G L

1978-01-01

Bacterial counts obtained on hydrophobic grid-membrane filters were comparable to conventional plate counts for Pseudomonas aeruginosa, Escherichia coli, and Staphylococcus aureus in homogenates from a range of foods. The wide numerical operating range of the hydrophobic grid-membrane filters allowed sequential diluting to be reduced or even eliminated, making them attractive as components in automated systems of analysis. Food debris could be rinsed completely from the unincubated hydrophobic grid-membrane filter surface without affecting the subsequent count, thus eliminating the possibility of counting food particles, a common source of error in electronic counting systems. PMID:100054

Bare below elbows: does this policy affect handwashing efficacy and reduce bacterial colonisation?

PubMed Central

Burger, A; Wijewardena, C; Clayson, S; Greatorex, RA

2010-01-01

INTRODUCTION UK Department of Health guidelines recommend that clinical staff are ‘bare below the elbows’. There is a paucity of evidence to support this policy. One may hypothesise that absence of clothing around wrists facilitates more effective handwashing: this study aims to establish whether dress code affects bacterial colonisation before and after handwashing. SUBJECTS AND METHODS Sixty-six clinical staff volunteered to take part in the study, noting whether they were bare below the elbows (BBE) or not bare (NB). Using a standardised technique, imprints of left and right fingers, palms, wrists and forearms were taken onto mini agar plates. Imprints were repeated after handwashing. After incubation, colonies per plate were counted, and subcultures taken. RESULTS Thirty-eight staff were BBE and 28 were not. A total of 1112 plates were cultured. Before handwashing there was no significant difference in number of colonies between BBE and NB groups (Mann–Whitney, P < 0.05). Handwashing reduced the colony count, with greatest effect on fingers, palms and dominant wrists (t-test, P < 0.05). Comparing the two groups again after handwashing revealed no significant difference (Mann–Whitney, P < 0.05). Subcultures revealed predominantly skin flora. CONCLUSIONS There was a large variation in number of colonies cultured. Handwashing resulted in a statistically significant reduction in colony count on fingers, palms and dominant wrist regardless of clothing. We conclude that handwashing produces a significant reduction in number of bacterial colonies on staff hands, and that clothing that is not BBE does not impede this reduction. PMID:20727253

Rapid assessment of viable but non-culturable Bacillus coagulans MTCC 5856 in commercial formulations using Flow cytometry.

PubMed

Majeed, Muhammed; Majeed, Shaheen; Nagabhushanam, Kalyanam; Punnapuzha, Ardra; Philip, Sheena; Mundkur, Lakshmi

2018-01-01

Accurate enumeration of bacterial count in probiotic formulation is imperative to ensure that the product adheres to regulatory standards and citation in consumer product label. Standard methods like plate count, can enumerate only replicating bacterial population under selected culture conditions. Viable but non culturable bacteria (VBNC) retain characteristics of living cells and can regain cultivability by a process known as resuscitation. This is a protective mechanism adapted by bacteria to evade stressful environmental conditions. B. coagulans MTCC 5856(LactoSpore®) is a probiotic endospore which can survive for decades in hostile environments without dividing. In the present study, we explored the use of flow cytometry to enumerate the viable count of B. coagulans MTCC 5856 under acidic and alkaline conditions, high temperature and in commercial formulations like compressed tablets and capsules. Flow cytometry (FCM) was comparable to plate count method when the spores were counted at physiological conditions. We show that VBNC state is induced in B. coagulans MTCC 5856by high temperature and acidic pH. The cells get resuscitated under physiological conditions and FCM was sensitive to detect the VBNC spores. Flow cytometry showed excellent ability to assess the viable spore count in commercial probiotic formulations of B. coagulans MTCC 5856. The results establish Flow cytometry as a reliable method to count viable bacteria in commercial probiotic preparations. Sporulation as well as existence as VBNC could contribute to the extreme stability of B. coagulans MTCC 5856.

Rapid assessment of viable but non-culturable Bacillus coagulans MTCC 5856 in commercial formulations using Flow cytometry

PubMed Central

Majeed, Muhammed; Majeed, Shaheen; Nagabhushanam, Kalyanam; Punnapuzha, Ardra; Philip, Sheena

2018-01-01

Accurate enumeration of bacterial count in probiotic formulation is imperative to ensure that the product adheres to regulatory standards and citation in consumer product label. Standard methods like plate count, can enumerate only replicating bacterial population under selected culture conditions. Viable but non culturable bacteria (VBNC) retain characteristics of living cells and can regain cultivability by a process known as resuscitation. This is a protective mechanism adapted by bacteria to evade stressful environmental conditions. B. coagulans MTCC 5856(LactoSpore®) is a probiotic endospore which can survive for decades in hostile environments without dividing. In the present study, we explored the use of flow cytometry to enumerate the viable count of B. coagulans MTCC 5856 under acidic and alkaline conditions, high temperature and in commercial formulations like compressed tablets and capsules. Flow cytometry (FCM) was comparable to plate count method when the spores were counted at physiological conditions. We show that VBNC state is induced in B. coagulans MTCC 5856by high temperature and acidic pH. The cells get resuscitated under physiological conditions and FCM was sensitive to detect the VBNC spores. Flow cytometry showed excellent ability to assess the viable spore count in commercial probiotic formulations of B. coagulans MTCC 5856. The results establish Flow cytometry as a reliable method to count viable bacteria in commercial probiotic preparations. Sporulation as well as existence as VBNC could contribute to the extreme stability of B. coagulans MTCC 5856. PMID:29474436

A rapid and universal bacteria-counting approach using CdSe/ZnS/SiO2 composite nanoparticles as fluorescence probe.

PubMed

Fu, Xin; Huang, Kelong; Liu, Suqin

2010-02-01

In this paper, a rapid, simple, and sensitive method was described for detection of the total bacterial count using SiO(2)-coated CdSe/ZnS quantum dots (QDs) as a fluorescence marker that covalently coupled with bacteria using glutaraldehyde as the crosslinker. Highly luminescent CdSe/ZnS were prepared by applying cadmium oxide and zinc stearate as precursors instead of pyrophoric organometallic precursors. A reverse-microemulsion technique was used to synthesize CdSe/ZnS/SiO(2) composite nanoparticles with a SiO(2) surface coating. Our results showed that CdSe/ZnS/SiO(2) composite nanoparticles prepared with this method possessed highly luminescent, biologically functional, and monodispersive characteristics, and could successfully be covalently conjugated with the bacteria. As a demonstration, it was found that the method had higher sensitivity and could count bacteria in 3 x 10(2) CFU/mL, lower than the conventional plate counting and organic dye-based method. A linear relationship of the fluorescence peak intensity (Y) and the total bacterial count (X) was established in the range of 3 x 10(2)-10(7) CFU/mL using the equation Y = 374.82X-938.27 (R = 0.99574). The results of the determination for the total count of bacteria in seven real samples were identical with the conventional plate count method, and the standard deviation was satisfactory.

A comparison of legionella and other bacteria concentrations in cooling tower water

DOE Office of Scientific and Technical Information (OSTI.GOV)

Cappabianca, R.M.; Jurinski, N.B.; Jurinski, J.B.

1994-05-01

A field study was conducted in which water samples collected from air conditioning cooling water reservoirs of high-rise buildings throughout an urban area were assayed for Legionella and for total bacteria. Buildings included within the study had ongoing biocidal treatment programs for the cooling towers. Separate sample analyses were performed to measure the viable colony concentrations of total bacteria and of Legionella in the process waters. The occurrence and viable counts of Legionella in 304 environmental water samples were determined by inoculating them onto plates of buffered charcoal yeast extract (BCYE) agar medium (a presumptive screening method). The samples weremore » collected during summer months between July and September. BCYE plate cultures of 50 (16.4%) of the samples yielded Legionella with viable counts ranging from 2 to 608 colony forming units per milliliter. In the water samples, 281 (92.4%) yielded viable counts of bacteria that ranged from 9 to 1.2 x 10{sup 6} per milliliter. This study demonstrates that Legionella are commonly present in the water of air conditioning cooling towers and that there is no significant correlation between concurrently sampled culture plate counts of Legionella and total bacteria plate counts. Correspondingly, there is no demonstrated validity for use of total bacterial counts as an inferential surrogate for the concentration of Legionella in the water. 19 refs., 3 figs., 1 tab.« less

Correlation between mastitis occurrence and the count of microorganisms in bulk raw milk of bovine dairy herds in four selective culture media.

PubMed

Souto, Luís I M; Minagawa, Clarice Y; Telles, Evelise O; Garbuglio, Márcio A; Amaku, Marcos; Melville, Priscilla A; Dias, Ricardo A; Sakata, Sonia T; Benites, Nilson R

2010-02-01

Milk is the normal secretion of the mammary gland, practically free of colostrum and obtained by the complete milking of one or more healthy animals. Mastitis is an inflammatory process of the mammary gland and it may cause alterations in the milk. The present work aimed to verify whether it is possible, by means of the counts of microorganism in the bulk raw milk in four selective culture media, to establish a correlation with the occurrence of mastitis and therefore, to monitor this disease in bovine dairy herds. The following selective culture media were used: KF Streptococcus Agar, Edwards Agar, Baird-Parker Agar, Blood Agar plus potassium tellurite. Spearman's correlation coefficient was calculated in order to compare the occurrence of mastitis (percentage) in each herd with respective selective culture media counts of microorganisms in bulk raw milk. Thirty-six possibilities were analysed (Tamis and CMT-positive rates were compared with the log-transformed count in four selective culture media) and there was a negative correlation between Tamis 3 and the Baird-Parker Agar plate count. The total results of microbiological tests showed that there were three correlations of the counts in selective culture media. Fifty-two possibilities were analysed and there was a negative correlation between no-bacterial-growth mastitis rates and log10 of KF Streptoccocus Agar plate count and there were two positive correlations between coagulase-positive staphylococci and log10 of Baird-Parker Agar plate count and Blood Agar plus potassium tellurite plate count.

Okara: A Nutritionally Valuable By-product Able to Stabilize Lactobacillus plantarum during Freeze-drying, Spray-drying, and Storage.

PubMed

Quintana, Gabriel; Gerbino, Esteban; Gómez-Zavaglia, Andrea

2017-01-01

Okara is a nutritionally valuable by-product produced in large quantities as result of soymilk elaboration. This work proposes its use as both culture and dehydration medium during freeze-drying, spray-drying, and storage of Lactobacillus plantarum CIDCA 83114. Whole and defatted okara were employed as culture media for L. plantarum CIDCA 83114. The growth kinetics were followed by plate counting and compared with those of bacteria grown in MRS broth (control). No significant differences in plate counting were observed in the three media. The fatty acid composition of bacteria grown in whole and defatted okara showed a noticeable increase in the unsaturated/saturated (U/S) fatty acid ratio, with regard to bacteria grown in MRS. This change was mainly due to the increase in polyunsaturated fatty acids, namely C18:2. For dehydration assays, cultures in the stationary phase were neutralized and freeze-dried (with or without the addition of 250 mM sucrose) or spray-dried. Bacteria were plate counted immediately after freeze-drying or spray-drying and during storage at 4°C for 90 days. Freeze-drying in whole okara conducted to the highest bacterial recovery. Regarding storage, spray-dried bacteria previously grown in whole and defatted okara showed higher plate counts than those grown in MRS. On the contrary, freeze-dried bacteria previously grown in all the three culture media were those with the lowest plate counts. The addition of sucrose to the dehydration media improved their recovery. The higher recovery of microorganisms grown in okara after freeze-drying and spray-drying processes and during storage was ascribed to both the presence of fiber and proteins in the dehydration media, and the increase in U/S fatty acids ratio in bacterial membranes. The obtained results support for the first time the use of okara as an innovative matrix to deliver L. plantarum . Considering that okara is an agro-waste obtained in large quantities, these results represent an innovative strategy to add it value, providing a symbiotic ingredient with promising industrial applications in the development of novel functional foods and feeds.

Okara: A Nutritionally Valuable By-product Able to Stabilize Lactobacillus plantarum during Freeze-drying, Spray-drying, and Storage

PubMed Central

Quintana, Gabriel; Gerbino, Esteban; Gómez-Zavaglia, Andrea

2017-01-01

Okara is a nutritionally valuable by-product produced in large quantities as result of soymilk elaboration. This work proposes its use as both culture and dehydration medium during freeze-drying, spray-drying, and storage of Lactobacillus plantarum CIDCA 83114. Whole and defatted okara were employed as culture media for L. plantarum CIDCA 83114. The growth kinetics were followed by plate counting and compared with those of bacteria grown in MRS broth (control). No significant differences in plate counting were observed in the three media. The fatty acid composition of bacteria grown in whole and defatted okara showed a noticeable increase in the unsaturated/saturated (U/S) fatty acid ratio, with regard to bacteria grown in MRS. This change was mainly due to the increase in polyunsaturated fatty acids, namely C18:2. For dehydration assays, cultures in the stationary phase were neutralized and freeze-dried (with or without the addition of 250 mM sucrose) or spray-dried. Bacteria were plate counted immediately after freeze-drying or spray-drying and during storage at 4°C for 90 days. Freeze-drying in whole okara conducted to the highest bacterial recovery. Regarding storage, spray-dried bacteria previously grown in whole and defatted okara showed higher plate counts than those grown in MRS. On the contrary, freeze-dried bacteria previously grown in all the three culture media were those with the lowest plate counts. The addition of sucrose to the dehydration media improved their recovery. The higher recovery of microorganisms grown in okara after freeze-drying and spray-drying processes and during storage was ascribed to both the presence of fiber and proteins in the dehydration media, and the increase in U/S fatty acids ratio in bacterial membranes. The obtained results support for the first time the use of okara as an innovative matrix to deliver L. plantarum. Considering that okara is an agro-waste obtained in large quantities, these results represent an innovative strategy to add it value, providing a symbiotic ingredient with promising industrial applications in the development of novel functional foods and feeds. PMID:28446905

A comparison of methods to assess the antimicrobial activity of nanoparticle combinations on bacterial cells.

PubMed

Bankier, Claire; Cheong, Yuen; Mahalingam, Suntharavathanan; Edirisinghe, Mohan; Ren, Guogang; Cloutman-Green, Elaine; Ciric, Lena

2018-01-01

Bacterial cell quantification after exposure to antimicrobial compounds varies widely throughout industry and healthcare. Numerous methods are employed to quantify these antimicrobial effects. With increasing demand for new preventative methods for disease control, we aimed to compare and assess common analytical methods used to determine antimicrobial effects of novel nanoparticle combinations on two different pathogens. Plate counts of total viable cells, flow cytometry (LIVE/DEAD BacLight viability assay) and qPCR (viability qPCR) were used to assess the antimicrobial activity of engineered nanoparticle combinations (NPCs) on Gram-positive (Staphylococcus aureus) and Gram-negative (Pseudomonas aeruginosa) bacteria at different concentrations (0.05, 0.10 and 0.25 w/v%). Results were analysed using linear models to assess the effectiveness of different treatments. Strong antimicrobial effects of the three NPCs (AMNP0-2) on both pathogens could be quantified using the plate count method and flow cytometry. The plate count method showed a high log reduction (>8-log) for bacteria exposed to high NPC concentrations. We found similar antimicrobial results using the flow cytometry live/dead assay. Viability qPCR analysis of antimicrobial activity could not be quantified due to interference of NPCs with qPCR amplification. Flow cytometry was determined to be the best method to measure antimicrobial activity of the novel NPCs due to high-throughput, rapid and quantifiable results.

Beyond the standard plate count: genomic views into microbial food ecology

USDA-ARS?s Scientific Manuscript database

Food spoilage is a complex process that involves multiple species with specific niches and metabolic processes; bacterial culturing techniques are the traditional methods for identifying the microbes responsible. These culture-dependent methods may be considered selective, targeting the isolation of...

Diversity and dynamics of antibiotic-resistant bacteria in cheese as determined by PCR denaturing gradient gel electrophoresis.

PubMed

Flórez, Ana Belén; Mayo, Baltasar

2015-12-02

This work reports the composition and succession of tetracycline- and erythromycin-resistant bacterial communities in a model cheese, monitored by polymerase chain reaction denaturing gradient gel electrophoresis (PCR-DGGE). Bacterial 16S rRNA genes were examined using this technique to detect structural changes in the cheese microbiota over manufacturing and ripening. Total bacterial genomic DNA, used as a template, was extracted from cultivable bacteria grown without and with tetracycline or erythromycin (both at 25 μg ml(-1)) on a non-selective medium used for enumeration of total and viable cells (Plate Count agar with Milk; PCA-M), and from those grown on selective and/or differential agar media used for counting various bacterial groups; i.e., lactic acid bacteria (de Man, Rogosa and Sharpe agar; MRSA), micrococci and staphylococci (Baird-Parker agar; BPA), and enterobacteria (Violet Red Bile Glucose agar; VRBGA). Large numbers of tetracycline- and erythromycin-resistant bacteria were detected in cheese samples at all stages of ripening. Counts of antibiotic-resistant bacteria varied widely depending on the microbial group and the point of sampling. In general, resistant bacteria were 0.5-1.0 Log10 units fewer in number than the corresponding susceptible bacteria. The PCR-DGGE profiles obtained with DNA isolated from the plates for total bacteria and the different bacterial groups suggested Escherichia coli, Lactococcus lactis, Enterococcus faecalis and Staphylococcus spp. as the microbial types resistant to both antibiotics tested. This study shows the suitability of the PCR-DGGE technique for rapidly identifying and tracking antibiotic resistant populations in cheese and, by extension, in other foods. Copyright © 2015 Elsevier B.V. All rights reserved.

Accurate live and dead bacterial cell enumeration using flow cytometry (Conference Presentation)

NASA Astrophysics Data System (ADS)

Ou, Fang; McGoverin, Cushla; Swift, Simon; Vanholsbeeck, Frédérique

2017-03-01

Flow cytometry (FCM) is based on the detection of scattered light and fluorescence to identify cells with particular characteristics of interest. However most FCM cannot precisely control the flow through its interrogation point and hence the volume and concentration of the sample cannot be immediately obtained. The easiest, most reliable and inexpensive way of obtaining absolute counts with FCM is by using reference beads. We investigated a method of using FCM with reference beads to measure live and dead bacterial concentration over the range of 106 to 108 cells/mL and ratio varying from 0 to 100%. We believe we are the first to use this method for such a large cell concentration range while also establishing the effect of varying the live/dead bacteria ratios. Escherichia coli solutions with differing ratios of live:dead cells were stained with fluorescent dyes SYTO 9 and propidium iodide (PI), which label live and dead cells, respectively. Samples were measured using a LSR II Flow Cytometer (BD Biosciences); using 488 nm excitation with 20 mW power. Both SYTO 9 and PI fluorescence were collected and threshold was set to side scatter. Traditional culture-based plate count was done in parallel to the FCM analysis. The concentration of live bacteria from FCM was compared to that obtained by plate counts. Preliminary results show that the concentration of live bacteria obtained by FCM and plate counts correlate well with each other and indicates this may be extended to a wider concentration range or for studying other cell characteristics.

Microcoupon Assay Of Adhesion And Growth Of Bacterial Films

NASA Technical Reports Server (NTRS)

Pierson, Duane L.; Koenig, David W.

1994-01-01

Microbiological assay technique facilitates determination of some characteristics of sessile bacteria like those that attach to and coat interior walls of water-purification systems. Biofilms cause sickness and interfere with purification process. Technique enables direct measurement of rate of attachment of bacterial cells, their metabolism, and effects of chemicals on them. Used to quantify effects of both bactericides and growth-stimulating agents and in place of older standard plate-count and tube-dilution techniques.

Disposable bioluminescence-based biosensor for detection of bacterial count in food.

PubMed

Luo, Jinping; Liu, Xiaohong; Tian, Qing; Yue, Weiwei; Zeng, Jing; Chen, Guangquan; Cai, Xinxia

2009-11-01

A biosensor for rapid detection of bacterial count based on adenosine 5'-triphosphate (ATP) bioluminescence has been developed. The biosensor is composed of a key sensitive element and a photomultiplier tube used as a detector element. The disposable sensitive element consists of a sampler, a cartridge where intracellular ATP is chemically extracted from bacteria, and a microtube where the extracted ATP reacts with the luciferin-luciferase reagent to produce bioluminescence. The bioluminescence signal is transformed into relevant electrical signal by the detector and further measured with a homemade luminometer. Parameters affecting the amount of the extracted ATP, including the types of ATP extractants, the concentrations of ATP extractant, and the relevant neutralizing reagent, were optimized. Under the optimal experimental conditions, the biosensor showed a linear response to standard bacteria in a concentration range from 10(3) to 10(8) colony-forming units (CFU) per milliliter with a correlation coefficient of 0.925 (n=22) within 5min. Moreover, the bacterial count of real food samples obtained by the biosensor correlated well with those by the conventional plate count method. The proposed biosensor, with characteristics of low cost, easy operation, and fast response, provides potential application to rapid evaluation of bacterial contamination in the food industry, environment monitoring, and other fields.

PCR-DGGE analysis of bacterial community dynamics in kava beverages during refrigeration.

PubMed

Dong, J; Kandukuru, P; Huang, A S; Li, Y

2011-07-01

Kava beverages are highly perishable even under refrigerated conditions. This study aimed to investigate the bacterial community dynamics in kava beverages during refrigeration.  Four freshly made kava beverages were obtained from kava bars and stored at 4°C. On days 0, 3 and 6, the aerobic plate count (APC), lactic acid bacteria (LAB) count and yeast and mould count (YMC) of the samples were determined. Meanwhile, bacterial DNA was extracted from each sample and subjected to the polymerase chain reaction-denaturing gradient gel electrophoresis (PCR-DGGE). Moreover, species-specific PCR assays were employed to identify predominant Pseudomonas spp. involved in kava spoilage. Over the storage period, the APC, LAB count and YMC of the four kava beverages all increased, whereas their pH values decreased. The DGGE profile revealed diverse bacterial populations in the samples. LAB, such as Weissella soli, Lactobacillus spp. and Lactococcus lactis, were found in the kava beverages. Species-specific PCR assays detected Pseudomonas putida and Pseudomonas fluorescens in the samples; Ps. fluorescens became dominant during refrigeration. LAB and Pseudomonas may play a significant role in the spoilage of kava beverages. This study provides important information that may be used to extend the shelf life of kava beverages. © 2011 The Authors. Letters in Applied Microbiology © 2011 The Society for Applied Microbiology.

A field study evaluation of Petrifilm™ plates as a 24-h rapid diagnostic test for clinical mastitis on a dairy farm.

PubMed

Mansion-de Vries, Elisabeth Maria; Knorr, Nicole; Paduch, Jan-Hendrik; Zinke, Claudia; Hoedemaker, Martina; Krömker, Volker

2014-03-01

Clinical mastitis is one of the most common and expensive diseases of dairy cattle. To make an informed treatment decision, it is important to know the causative pathogen. However, no detection of bacterial growth can be made in approximately 30% of all clinical cases of mastitis. Before selecting the treatment regimen, it is important to know whether the mastitis-causing pathogen (MCP) is Gram-positive or Gram-negative. The aim of this field study was to investigate whether using two 3M Petrifilm™ products on-farm (which conveys a higher degree of sample freshness but also bears a higher risk for contamination than working in a lab) as 24-h rapid diagnostic of clinical mastitis achieved results that were comparable to the conventional microbiological diagnostic method. AerobicCount (AC)-Petrifilm™ and ColiformCount (CC)-Petrifilm™ were used to identify the total bacterial counts and Gram-negative bacteria in samples from clinical mastitis cases, respectively. Missing growth on both plates was classified as no bacterial detection. Growth only on the AC-Petrifilm™ was assessed as Gram-positive, and growth on both Petrifilm™ plates was assessed as Gram-negative bacterial growth. Additionally, milk samples were analysed by conventional microbiological diagnostic method on aesculin blood agar as a reference method. Overall, 616 samples from clinical mastitis cases were analysed. Using the reference method, Gram-positive and Gram-negative bacteria, mixed bacterial growth, contaminated samples and yeast were determined in 32.6%, 20.0%, 2.5%, 14.1% and 1.1% of the samples, respectively. In 29.7% of the samples, microbiological growth could not be identified. Using the Petrifilm™ concept, bacterial growth was detected in 59% of the culture-negative samples. The sensitivity of the Petrifilm™ for Gram-positive and Gram-negative MCP was 85.2% and 89.9%, respectively. The specificity was 75.4% for Gram-positive and 88.4% for Gram-negative MCP. For the culture-negative samples, sensitivity was 41.0% and specificity was 91.0%. The results indicate that the Petrifilm™ concept is suitable for therapeutic decision-making at the farm level or in veterinary practice. As this concept does not allow any statement about the genus or species of microorganisms, relevant MCP should be assessed periodically at the herd level with conventional microbiological diagnostics. Copyright © 2014 Elsevier B.V. All rights reserved.

Assessment of the bacteriological activity associated with granular activated carbon treatment of drinking water.

PubMed Central

Stewart, M H; Wolfe, R L; Means, E G

1990-01-01

Bacteriological analyses were performed on the effluent from a conventional water treatment pilot plant in which granular activated carbon (GAC) had been used as the final process to assess the impact of GAC on the microbial quality of the water produced. Samples were collected twice weekly for 160 days from the effluents of six GAC columns, each of which used one of four different empty-bed contact times (7.5, 15, 30, and 60 min). The samples were analyzed for heterotrophic plate counts and total coliforms. Effluent samples were also exposed to chloramines and free chlorine for 60 min (pH 8.2, 23 degrees C). Bacterial identifications were performed on the disinfected and nondisinfected effluents. Additional studies were conducted to assess the bacteriological activity associated with released GAC particles. The results indicated that heterotrophic plate counts in the effluents from all columns increased to 10(5) CFU/ml within 5 days and subsequently stabilized at 10(4) CFU/ml. The heterotrophic plate counts did not differ at different empty-bed contact times. Coliforms (identified as Enterobacter spp.) were recovered from the nondisinfected effluent on only two occasions. The disinfection results indicated that 1.5 mg of chloramines per liter inactivated approximately 50% more bacteria than did 1.0 mg of free chlorine per liter after 1 h of contact time. Chloramines and chlorine selected for the development of different bacterial species--Pseudomonas spp. and Flavobacterium spp., respectively.(ABSTRACT TRUNCATED AT 250 WORDS) PMID:2082828

Evaluation of the effects of footwear hygiene protocols on nonspecific bacterial contamination of floor surfaces in an equine hospital.

PubMed

Stockton, Kelly A; Morley, Paul S; Hyatt, Doreene R; Burgess, Brandy A; Patterson, Gage; Dunowska, Magda; Lee, David E

2006-04-01

To evaluate the effects of footwear hygiene protocols on bacterial contamination of floor surfaces in an equine hospital. Field trial. Footwear hygiene protocols evaluated included use of rubber overboots with footbaths and footmats containing a quaternary ammonium disinfectant, rubber overboots with footbaths and footmats containing a peroxygen disinfectant, and no restrictions on footwear type but mandatory use of footbaths and footmats containing a peroxygen disinfectant. Nonspecific aerobic bacterial counts were determined via 2 procedures for sample collection and bacterial enumeration (contact plates vs swabbing combined with use of spread plates), and the effects of each footwear hygiene protocol were compared. There were no consistent findings suggesting that any of the protocols were associated with differences in numbers of bacteria recovered from floor surfaces. Although there were detectable differences in numbers of bacteria recovered in association with different footwear hygiene protocols, differences in least square mean bacterial counts did not appear to be clinically relevant (ie, were < 1 log10). Although cleaning and disinfection of footwear are important aids in reducing the risk of nosocomial transmission of infectious agents in veterinary hospitals, the numbers of aerobic bacteria recovered from floor surfaces were not affected by use of rubber overboots or the types of disinfectant used in this study. Further study is warranted to evaluate the usefulness of footwear hygiene practices relative to their efficacy for reducing transmission of specific pathogens or decreasing nosocomial disease risk.

Biodegradable Chitosan Coating Incorporated with Black Pepper Essential Oil for Shelf Life Extension of Common Carp (Cyprinus carpio) during Refrigerated Storage.

PubMed

Moosavi-Nasab, Marzieh; Shad, Ehsan; Ziaee, Esmaeil; Yousefabad, Seyyed Hossein Asadi; Golmakani, Mohammad Taghi; Azizinia, Mehdi

2016-06-01

Chitosan (Ch) coating incorporated with black pepper essential oil (Ch+BPEO) was studied to extend the shelf life of common carp (Cyprinus carpio) during refrigerated storage at 4 ± 1°C. The chemical composition of BPEO was characterized using gas chromatography-mass spectrometry (GC-MS). Antibacterial properties of BPEO were determined by disk diffusion agar, MIC, and MBC. Ch (2% [wt/vol]) and Ch+BPEO (2% [wt/vol] Ch with 1.5% [vol/vol] BPEO) were used for common carp fillet coating. The samples were analyzed periodically for chemical (pH, total volatile basic nitrogen) and microbiological (aerobic plate count, psychrophilic bacteria count, lactic acid bacteria, and Enterobacteriaceae bacterial counts) characteristics during 16 days. The GC-MS results indicated that main components in BPEO were carene, caryophyllene, limonene, β-pinene, and α-pinene. The samples coated with Ch and Ch+BPEO resulted in lower pH and total volatile basic nitrogen values in comparison with the control. The microbiological analysis of fish fillets during refrigerated storage clearly indicated that Ch+BPEO coating significantly reduced the fish fillet microbial load. The aerobic plate count, psychrophilic bacteria count, lactic acid bacteria count, and Enterobacteriaceae bacterial count of samples coated with Ch+BPEO were reduced approximately 4.1, 3.9, 2.3, and 2.8 log CFU/g, respectively, at the end of the storage period. Finally, Ch and Ch+BPEO effectively improved the quality of fish fillet during refrigerated storage and extended the shelf life of fish fillets from 8 to 16 days. Black pepper; Chitosan; Common carp; Essential oil.

The degree of bacterial contamination while performing spine surgery.

PubMed

Ahn, Dong Ki; Park, Hoon Seok; Kim, Tae Woo; Yang, Jong Hwa; Boo, Kyung Hwan; Kim, In Ja; Lee, Hye Jin

2013-03-01

Prospective experimental study. To evaluate bacterial contamination during surgery. The participants of surgery and ventilation system have been known as the most significant sources of contamination. Two pairs of air culture blood agar plate for G(+) bacteria and MacConkey agar plate for G(-) bacteria were placed at 3 different locations in a conventional operation room: in the surgical field, under the airflow of local air conditioner, and pathway to door while performing spine surgeries. One pair of culture plates was retrieved after one hour and the other pair was retrieved after 3 hours. The cultured bacteria were identified and number of colonies was counted. There was no G(-) bacteria identified. G(+) bacteria grew on all 90 air culture blood agar plates. The colony count of one hour group was 14.5±5.4 in the surgical field, 11.3±6.6 under the local air conditioner, and 13.1±8.7 at the pathway to the door. There was no difference among the 3 locations. The colony count of 3 hours group was 46.4±19.5, 30.3±12.9, and 39.7±15.2, respectively. It was more at the surgical field than under the air conditioner (p=0.03). The number of colonies of one hour group was 13.0±7.0 and 3 hours group was 38.8±17.1. There was positive correlation between the time and the number of colonies (r=0.76, p=0.000). Conventional operation room was contaminated by G(+) bacteria. The degree of contamination was most high at the surgical field. The number of bacteria increased right proportionally to the time.

The Degree of Bacterial Contamination While Performing Spine Surgery

PubMed Central

Ahn, Dong Ki; Park, Hoon Seok; Yang, Jong Hwa; Boo, Kyung Hwan; Kim, In Ja; Lee, Hye Jin

2013-01-01

Study Design Prospective experimental study. Purpose To evaluate bacterial contamination during surgery. Overview of Literature The participants of surgery and ventilation system have been known as the most significant sources of contamination. Methods Two pairs of air culture blood agar plate for G(+) bacteria and MacConkey agar plate for G(-) bacteria were placed at 3 different locations in a conventional operation room: in the surgical field, under the airflow of local air conditioner, and pathway to door while performing spine surgeries. One pair of culture plates was retrieved after one hour and the other pair was retrieved after 3 hours. The cultured bacteria were identified and number of colonies was counted. Results There was no G(-) bacteria identified. G(+) bacteria grew on all 90 air culture blood agar plates. The colony count of one hour group was 14.5±5.4 in the surgical field, 11.3±6.6 under the local air conditioner, and 13.1±8.7 at the pathway to the door. There was no difference among the 3 locations. The colony count of 3 hours group was 46.4±19.5, 30.3±12.9, and 39.7±15.2, respectively. It was more at the surgical field than under the air conditioner (p=0.03). The number of colonies of one hour group was 13.0±7.0 and 3 hours group was 38.8±17.1. There was positive correlation between the time and the number of colonies (r=0.76, p=0.000). Conclusions Conventional operation room was contaminated by G(+) bacteria. The degree of contamination was most high at the surgical field. The number of bacteria increased right proportionally to the time. PMID:23508998

Microbiological and molecular characterization of commercially available probiotics containing Bacillus clausii from India and Pakistan.

PubMed

Patrone, Vania; Molinari, Paola; Morelli, Lorenzo

2016-11-21

Probiotics are actively used for treatment of diarrhoea, respiratory infections, and prevention of infectious gastrointestinal diseases. The efficacy of probiotics is due to strain-specific features and the number of viable cells; however, several reports of deviations from the label in the actual content of strains in probiotic products are a matter of concern. Most of the available data on quality focuses on probiotic products containing lactobacilli and/or bifidobacteria, while very few data are available on spore-forming probiotics. The present study evaluates the label claims for spore count and species identification in five commercial probiotic products marketed in India and Pakistan that claim to contain Bacillus clausii: Tufpro, Ecogro, Enterogermina, Entromax, and Ospor. Bacterial enumeration from three batches was done by microbiological plating methods by two independent operators. Species identification was done using PCR amplification and sequence analysis of the 16S rRNA gene, and determination of the total amount of species present in the products was done using PCR-denaturing gradient gel electrophoresis (PCR-DGGE) analysis followed by DNA sequencing of the excised bands. Plate count methods demonstrated poor correlations between quantitative label indications and bacteria recovered from plates for Tufpro, Ecogro, and Ospor. The 16S rRNA analysis performed on bacteria isolated from plate counts showed that only Enterogermina and Ospor contained homogenous B. clausii. PCR-DGGE analysis revealed that only Enterogermina had a homogenous B. clausii population while other products had mixed bacterial populations. In conclusion, the current analysis clearly demonstrates that of the five analysed commercial probiotics, only Enterogermina followed the label claims. Copyright © 2016 Elsevier B.V. All rights reserved.

EFFECT OF AEROSOLIZATION ON CULTURABILITY AND VIABILITY OF GRAM-NEGATIVE BACTERIA

EPA Science Inventory

Estimations of the bacterial content of air can be more easily made now than a decade ago, with colony formation the method of choice for enumeration of airborne bacteria.However, plate counts are subject to error because bacteria exposed to the air may remain viable yet lose the...

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.

MRP8/14 Enhances Corneal Susceptibility to Pseudomonas aeruginosa Infection by Amplifying Inflammatory Responses

PubMed Central

Deng, Qiuchan; Sun, Mingxia; Yang, Kun; Zhu, Min; Chen, Kang; Yuan, Jin; Wu, Minhao; Huang, Xi

2013-01-01

Purpose. We explored the role of myeloid-related protein 8 and 14 (MRP8/14) in Pseudomonas aeruginosa (PA) keratitis. Methods. MRP8/14 mRNA levels in human corneal scrapes and mouse corneas infected by PA were tested using real-time PCR. MRP8/14 protein expression in C57BL/6 (B6) corneas was confirmed using Western blot assay and immunohistochemistry. B6 mice were injected subconjunctivally with siRNA for MRP8/14, and then infected with PA. Bacterial plate counts and myeloperoxidase assays were used to determine the bacterial load and polymorphonuclear neutrophil (PMN) infiltration in infected B6 corneas. Pro-inflammatory cytokine levels in vivo and in vitro were examined with PCR and ELISA. In murine macrophage-like RAW264.7 cells, phagocytosis and bacterial killing were assessed using plate count assays, and reactive oxygen species (ROS) and nitric oxide (NO) levels were tested with flow cytometry and Griess assay, respectively. Results. MRP8/14 expression levels were increased significantly in human corneal scrapes and B6 corneas after PA infection. Silencing of MRP8/14 in B6 corneas significantly reduced the severity of corneal disease, bacterial clearance, PMN infiltration, and pro-inflammatory cytokine expression after PA infection. In vitro studies demonstrated further that silencing of MRP8/14 suppressed pro-inflammatory cytokine production, bacterial killing, and ROS production, but not phagocytosis or NO production. Conclusions. Our study demonstrated a dual role for MRP8/14 in bacterial keratitis. Although MRP8/14 promotes bacterial clearance by enhancing ROS production, it functions more importantly as an inflammatory amplifier at the ocular surface by enhancing pro-inflammatory cytokine expression, thus contributing to the corneal susceptibility. PMID:23299480

Long-Term Bacterial Dynamics in a Full-Scale Drinking Water Distribution System

PubMed Central

Prest, E. I.; Weissbrodt, D. G.; Hammes, F.; van Loosdrecht, M. C. M.; Vrouwenvelder, J. S.

2016-01-01

Large seasonal variations in microbial drinking water quality can occur in distribution networks, but are often not taken into account when evaluating results from short-term water sampling campaigns. Temporal dynamics in bacterial community characteristics were investigated during a two-year drinking water monitoring campaign in a full-scale distribution system operating without detectable disinfectant residual. A total of 368 water samples were collected on a biweekly basis at the water treatment plant (WTP) effluent and at one fixed location in the drinking water distribution network (NET). The samples were analysed for heterotrophic plate counts (HPC), Aeromonas plate counts, adenosine-tri-phosphate (ATP) concentrations, and flow cytometric (FCM) total and intact cell counts (TCC, ICC), water temperature, pH, conductivity, total organic carbon (TOC) and assimilable organic carbon (AOC). Multivariate analysis of the large dataset was performed to explore correlative trends between microbial and environmental parameters. The WTP effluent displayed considerable seasonal variations in TCC (from 90 × 103 cells mL-1 in winter time up to 455 × 103 cells mL-1 in summer time) and in bacterial ATP concentrations (<1–3.6 ng L-1), which were congruent with water temperature variations. These fluctuations were not detected with HPC and Aeromonas counts. The water in the network was predominantly influenced by the characteristics of the WTP effluent. The increase in ICC between the WTP effluent and the network sampling location was small (34 × 103 cells mL-1 on average) compared to seasonal fluctuations in ICC in the WTP effluent. Interestingly, the extent of bacterial growth in the NET was inversely correlated to AOC concentrations in the WTP effluent (Pearson’s correlation factor r = -0.35), and positively correlated with water temperature (r = 0.49). Collecting a large dataset at high frequency over a two year period enabled the characterization of previously undocumented seasonal dynamics in the distribution network. Moreover, high-resolution FCM data enabled prediction of bacterial cell concentrations at specific water temperatures and time of year. The study highlights the need to systematically assess temporal fluctuations in parallel to spatial dynamics for individual drinking water distribution systems. PMID:27792739

Long-Term Bacterial Dynamics in a Full-Scale Drinking Water Distribution System.

PubMed

Prest, E I; Weissbrodt, D G; Hammes, F; van Loosdrecht, M C M; Vrouwenvelder, J S

2016-01-01

Large seasonal variations in microbial drinking water quality can occur in distribution networks, but are often not taken into account when evaluating results from short-term water sampling campaigns. Temporal dynamics in bacterial community characteristics were investigated during a two-year drinking water monitoring campaign in a full-scale distribution system operating without detectable disinfectant residual. A total of 368 water samples were collected on a biweekly basis at the water treatment plant (WTP) effluent and at one fixed location in the drinking water distribution network (NET). The samples were analysed for heterotrophic plate counts (HPC), Aeromonas plate counts, adenosine-tri-phosphate (ATP) concentrations, and flow cytometric (FCM) total and intact cell counts (TCC, ICC), water temperature, pH, conductivity, total organic carbon (TOC) and assimilable organic carbon (AOC). Multivariate analysis of the large dataset was performed to explore correlative trends between microbial and environmental parameters. The WTP effluent displayed considerable seasonal variations in TCC (from 90 × 103 cells mL-1 in winter time up to 455 × 103 cells mL-1 in summer time) and in bacterial ATP concentrations (<1-3.6 ng L-1), which were congruent with water temperature variations. These fluctuations were not detected with HPC and Aeromonas counts. The water in the network was predominantly influenced by the characteristics of the WTP effluent. The increase in ICC between the WTP effluent and the network sampling location was small (34 × 103 cells mL-1 on average) compared to seasonal fluctuations in ICC in the WTP effluent. Interestingly, the extent of bacterial growth in the NET was inversely correlated to AOC concentrations in the WTP effluent (Pearson's correlation factor r = -0.35), and positively correlated with water temperature (r = 0.49). Collecting a large dataset at high frequency over a two year period enabled the characterization of previously undocumented seasonal dynamics in the distribution network. Moreover, high-resolution FCM data enabled prediction of bacterial cell concentrations at specific water temperatures and time of year. The study highlights the need to systematically assess temporal fluctuations in parallel to spatial dynamics for individual drinking water distribution systems.

Bacteriocidal activity of sanitizers against Enterococcus faecium attached to stainless steel as determined by plate count and impedance methods.

PubMed

Andrade, N J; Bridgeman, T A; Zottola, E A

1998-07-01

Enterococcus faecium attached to stainless steel chips (100 mm2) was treated with the following sanitizers: sodium hypochlorite, peracetic acid (PA), peracetic acid plus an organic acid (PAS), quaternary ammonium, organic acid, and anionic acid. The effectiveness of sanitizer solutions on planktonic cells (not attached) was evaluated by the Association of Official Analytical Chemists (AOAC) suspension test. The number of attached cells was determined by impedance measurement and plate count method after vortexing. The decimal reduction (DR) in numbers of the E. faecium population was determined for the three methods and was analyzed by analysis of variance (P < 0.05) using Statview software. The adhered cells were more resistant (P < 0.05) than nonadherent cells. The DR averages for all of the sanitizers for 30 s of exposure were 6.4, 2.2, and 2.5 for the AOAC suspension test, plate count method after vortexing, and impedance measurement, respectively. Plate count and impedance methods showed a difference (P < 0.05) after 30 s of sanitizer exposure but not after 2 min. The impedance measurement was the best method to measure adherent cells. Impedance measurement required the development of a quadratic regression. The equation developed from 82 samples is as follows: log CFU/chip = 0.2385T2-0.96T + 9.35, r2 = 0.92, P < 0.05, T = impedance detection time in hours. This method showed that the sanitizers PAS and PA were more effective against E. faecium than the other sanitizers. At 30 s, the impedance method recovered about 25 times more cells than the plate count method after vortexing. These data suggest that impedance measurement is the method of choice when evaluating the number of bacterial cells adhered to a surface.

Rapid and direct detection of Invivo kinetics of pathogenic bacterial infection from mouse blood and urine.

PubMed

Gopal, Judy; Lee, Chia-Hsun; Wu, Hui-Fen

2012-06-06

This study demonstrates the first use of matrix assisted laser desorption/ionization mass spectrometry (MALDI-MS) to trace the Invivo infection kinetics of the well known deadly pathogen Staphylococcus aureus in Swiss albino mice. The growth curve of the bacteria from the point of injection (200μL of bacterial suspension (10(8)cfu/mL)) into the mouse blood till mortality (death) was periodically analyzed using the plate counting method and MALDI-MS. Bacterial counts of 10(3)cfu/mL were observed in the log phase of the growth curve in the blood and 10(2)cfu/mL were observed in the urine samples. Death occurred in the log phase of the growth curve, where the bacterial counts showed steady increase. In other cases, the bacteria counts started decreasing after 48h and by 96h the bacteria got totally eliminated from the mouse and these mice survived. Direct MALDI-MS was not feasible for tracking the bacteria in the infected blood. However, ionic liquid 1-Butyl-3-methylimidazolium tetrafluoroborate was successful in enabling bacterial detection amidst the strong blood peaks. But, in the case of the urine analysis, it was observed that direct MALDI-MS was adequate to enable detection. The results obtained prove the efficacy of MALDI-MS for analyzing pathogenic bacteria in clinical samples. This article is part of a Special Issue entitled: Proteomics: The clinical link. Copyright © 2011 Elsevier B.V. All rights reserved.

Bacterial, Fungal, and Actinomycete Populations in Soils Receiving Repeated Applications of 2,4-Dichlorophenoxyacetic Acid and Trifluralin 1

PubMed Central

Breazeale, F. W.; Camper, N. D.

1970-01-01

Soil samples were collected from an untreated plot and plots receiving repeated applications of 2,4-dichlorophenoxyacetic acid (2,4-D) and α,α,α-trifluoro-2, 6-dinitro-N,N-dipropyl-p-toluidine (trifluralin); they were then plated on media specific for bacteria, fungi, and actinomycetes. The actinomycete colony count in the trifluralin-treated plot was greater than the control, but the same as the control in the 2,4-D-treated plot. The bacterial count was lower in both treated plots. Fungal colonies in the trifluralin-treated plots were greater than the control, but not different from the control in the 2,4-D-treated plot. PMID:5437308

Pseudoalteromonas spp. Serve as Initial Bacterial Attractants in Mesocosms of Coastal Waters but Have Subsequent Antifouling Capacity in Mesocosms and when Embedded in Paint

PubMed Central

Bernbom, Nete; Ng, Yoke Yin; Olsen, Stefan Møller

2013-01-01

The purpose of the present study was to determine if the monoculture antifouling effect of several pigmented pseudoalteromonads was retained in in vitro mesocosm systems using natural coastal seawater and when the bacteria were embedded in paint used on surfaces submerged in coastal waters. Pseudoalteromonas piscicida survived on a steel surface and retained antifouling activity for at least 53 days in sterile seawater, whereas P. tunicata survived and had antifouling activity for only 1 week. However, during the first week, all Pseudoalteromonas strains facilitated rather than prevented bacterial attachment when used to coat stainless steel surfaces and submerged in mesocosms with natural seawater. The bacterial density on surfaces coated with sterile growth medium was 105 cells/cm2 after 7 days, whereas counts on surfaces precoated with Pseudoalteromonas were significantly higher, at 106 to 108 cells/cm2. However, after 53 days, seven of eight Pseudoalteromonas strains had reduced total bacterial adhesion compared to the control. P. piscicida, P. antarctica, and P. ulvae remained on the surface, at levels similar to those in the initial coating, whereas P. tunicata could not be detected. Larger fouling organisms were observed on all plates precoated with Pseudoalteromonas; however, plates coated only with sterile growth medium were dominated by a bacterial biofilm. Suspensions of a P. piscicida strain and a P. tunicata strain were incorporated into ship paints (Hempasil x3 87500 and Hempasil 77500) used on plates that were placed at the Hempel A/S test site in Jyllinge Harbor. For the first 4 months, no differences were observed between control plates and treated plates, but after 5 to 6 months, the control plates were more fouled than the plates with pseudoalteromonad-based paint. Our study demonstrates that no single laboratory assay can predict antifouling effects and that a combination of laboratory and real-life methods must be used to determine the potential antifouling capability of new agents or organisms. PMID:23995925

Evaluation of quantitative PCR measurement of bacterial colonization of epithelial cells.

PubMed

Schmidt, Marcin T; Olejnik-Schmidt, Agnieszka K; Myszka, Kamila; Borkowska, Monika; Grajek, Włodzimierz

2010-01-01

Microbial colonization is an important step in establishing pathogenic or probiotic relations to host cells and in biofilm formation on industrial or medical devices. The aim of this work was to verify the applicability of quantitative PCR (Real-Time PCR) to measure bacterial colonization of epithelial cells. Salmonella enterica and Caco-2 intestinal epithelial cell line was used as a model. To verify sensitivity of the assay a competition of the pathogen cells to probiotic microorganism was tested. The qPCR method was compared to plate count and radiolabel approach, which are well established techniques in this area of research. The three methods returned similar results. The best quantification accuracy had radiolabel method, followed by qPCR. The plate count results showed coefficient of variation two-times higher than this of qPCR. The quantitative PCR proved to be a reliable method for enumeration of microbes in colonization assay. It has several advantages that make it very useful in case of analyzing mixed populations, where several different species or even strains can be monitored at the same time.

A Microbial Community in Sediments Beneath the Western Antarctic Ice Sheet, Ice Stream C (Kamb)

NASA Astrophysics Data System (ADS)

Skidmore, M.; Han, S.; Foo, W.; Bui, D.; Lanoil, B.

2004-12-01

In 2000, an ice-drilling project focusing on the "sticky spot" of Ice Stream C recovered cores of sub-glacial sediments from beneath the Western Antarctic Ice Sheet. We have characterized several chemical and microbiological parameters of the sole intact sediment core. Pore waters extracted from these sediments were brackish and some were supersaturated with respect to calcite. Ion chromatography demonstrated the presence of several organic acids at low, but detectable, levels in the pore water. DAPI direct cell counts were approximately 107 cells g-1. Aerobic viable plate counts were much lower than direct cell counts; however, they were two orders of magnitude higher on plates incubated at low temperature (4 ° C; 3.63 x 105 CFU ml-1) than at higher temperatures (ca. 22° C; 1.5 x 103 CFU ml-1); no colonies were detected on plates incubated anaerobically at either temperature. 16S rDNA clone library analysis indicates extremely limited bacterial diversity in these samples: six phylogenetic clades were detected. The three dominant bacterial phylogenetic clades in the clone libraries (252 clones total) were most closely related to Thiobacillus thioparus (180 clones), Polaromonas vacuolata (34 clones), and Gallionella ferruginea (35 clones) and their relatives; one clone each represented the other three phylogenetic clades (most closely related to Ralstonia pickettii, Lysobacter antibioticus, and Xylella fastidiosa, respectively). These sequences match closely with sequences previously obtained from other subglacial environments in Alaska, Ellesmere Island, Canada and New Zealand. Implications of this microbial community to subglacial chemistry and microbial biogeography will be discussed.

Experimental Study for Automatic Colony Counting System Based Onimage Processing

NASA Astrophysics Data System (ADS)

Fang, Junlong; Li, Wenzhe; Wang, Guoxin

Colony counting in many colony experiments is detected by manual method at present, therefore it is difficult for man to execute the method quickly and accurately .A new automatic colony counting system was developed. Making use of image-processing technology, a study was made on the feasibility of distinguishing objectively white bacterial colonies from clear plates according to the RGB color theory. An optimal chromatic value was obtained based upon a lot of experiments on the distribution of the chromatic value. It has been proved that the method greatly improves the accuracy and efficiency of the colony counting and the counting result is not affected by using inoculation, shape or size of the colony. It is revealed that automatic detection of colony quantity using image-processing technology could be an effective way.

Bacteria killing nanotechnology Bio-Kil effectively reduces bacterial burden in intensive care units.

PubMed

Hsueh, P-R; Huang, H-C; Young, T-G; Su, C-Y; Liu, C-S; Yen, M-Y

2014-04-01

A contaminated hospital environment has been identified as an important reservoir of pathogens causing healthcare-associated infections. This study is to evaluate the efficacy of bacteria killing nanotechnology Bio-Kil on reducing bacterial counts in an intensive care unit (ICU). Two single-bed rooms (S-19 and S-20) in the ICU were selected from 7 April to 27 May 2011. Ten sets of new textiles (pillow cases, bed sheets, duvet cover, and patient clothing) used by patients in the two single-bed rooms were provided by the sponsors. In the room S-20, the 10 sets of new textiles were washed with Bio-Kil; the room walls, ceiling, and air-conditioning filters were treated with Bio-Kil; and the surfaces of instruments (respirator, telephone, and computer) were covered with Bio-Kil-embedded silicon pads. Room S-19 served as the control. We compared the bacterial count on textiles and environment surfaces as well as air samples between the two rooms. A total of 1,364 samples from 22 different sites in each room were collected. The mean bacterial count on textiles and environmental surfaces in room S-20 was significantly lower than that in room S-19 (10.4 vs 49.6 colony-forming units [CFU]/100 cm(2); P < 0.001). Room S-20 had lower bacterial counts in air samples than room S-19 (33.4-37.6 vs 21.6-25.7 CFU/hour/plate; P < 0.001). The density of microbial isolations was significantly greater among patients admitted to room S-19 than those to room S-20 (9.15 vs 5.88 isolates per 100 patient-days, P < 0.05). Bio-Kil can significantly reduce bacterial burden in the environment of the ICU.

Identification of Lactobacillus delbrueckii and Streptococcus thermophilus Strains Present in Artisanal Raw Cow Milk Cheese Using Real-time PCR and Classic Plate Count Methods.

PubMed

Stachelska, Milena A

2017-12-04

The aim of this paper was to detect Lactobacillus delbrueckii and Streptococcus thermophilus using real-time quantitative PCR assay in 7-day ripening cheese produced from unpasteurised milk. Real-time quantitative PCR assays were designed to identify and enumerate the chosen species of lactic acid bacteria (LAB) in ripened cheese. The results of molecular quantification and classic bacterial enumeration showed a high level of similarity proving that DNA extraction was carried out in a proper way and that genomic DNA solutions were free of PCR inhibitors. These methods revealed the presence of L. delbrueckii and S. thermophilus. The real-time PCR enabled quantification with a detection of 101-103 CFU/g of product. qPCR-standard curves were linear over seven log units down to 101 copies per reaction; efficiencies ranged from 77.9% to 93.6%. Cheese samples were analysed with plate count method and qPCR in parallel. Compared with the classic plate count method, the newly developed qPCR method provided faster and species specific identification of two dairy LAB and yielded comparable quantitative results.

Hydrazine vapor inactivates Bacillus spores

NASA Astrophysics Data System (ADS)

Schubert, Wayne W.; Engler, Diane L.; Beaudet, Robert A.

2016-05-01

NASA policy restricts the total number of bacterial spores that can remain on a spacecraft traveling to any planetary body which might harbor life or have evidence of past life. Hydrazine, N2H4, is commonly used as a propellant on spacecraft. Hydrazine as a liquid is known to inactivate bacterial spores. We have now verified that hydrazine vapor also inactivates bacterial spores. After Bacillus atrophaeus ATCC 9372 spores deposited on stainless steel coupons were exposed to saturated hydrazine vapor in closed containers, the spores were recovered from the coupons, serially diluted, pour plated and the surviving bacterial colonies were counted. The exposure times required to reduce the spore population by a factor of ten, known as the D-value, were 4.70 ± 0.50 h at 25 °C and 2.85 ± 0.13 h at 35 °C. These inactivation rates are short enough to ensure that the bioburden of the surfaces and volumes would be negligible after prolonged exposure to hydrazine vapor. Thus, all the propellant tubing and internal tank surfaces exposed to hydrazine vapor do not contribute to the total spore count.

Total and Viable Legionella pneumophila Cells in Hot and Natural Waters as Measured by Immunofluorescence-Based Assays and Solid-Phase Cytometry ▿†

PubMed Central

Parthuisot, N.; Binet, M.; Touron-Bodilis, A.; Pougnard, C.; Lebaron, P.; Baudart, J.

2011-01-01

A new method was developed for the rapid and sensitive detection of viable Legionella pneumophila. The method combines specific immunofluorescence (IF) staining using monoclonal antibodies with a bacterial viability marker (ChemChrome V6 cellular esterase activity marker) by means of solid-phase cytometry (SPC). IF methods were applied to the detection and enumeration of both the total and viable L. pneumophila cells in water samples. The sensitivity of the IF methods coupled to SPC was 34 cells liter−1, and the reproducibility was good, with the coefficient of variation generally falling below 30%. IF methods were applied to the enumeration of total and viable L. pneumophila cells in 46 domestic hot water samples as well as in cooling tower water and natural water samples, such as thermal spring water and freshwater samples. Comparison with standard plate counts showed that (i) the total direct counts were always higher than the plate counts and (ii) the viable counts were higher than or close to the plate counts. With domestic hot waters, when the IF assay was combined with the viability test, SPC detected up to 3.4 × 103 viable but nonculturable L. pneumophila cells per liter. These direct IF methods could be a powerful tool for high-frequency monitoring of domestic hot waters or for investigating the occurrence of viable L. pneumophila in both man-made water systems and environmental water samples. PMID:21742913

Smartphone-based rapid quantification of viable bacteria by single-cell microdroplet turbidity imaging.

PubMed

Cui, Xiaonan; Ren, Lihui; Shan, Yufei; Wang, Xixian; Yang, Zhenlong; Li, Chunyu; Xu, Jian; Ma, Bo

2018-05-18

Standard plate count (SPC) has been recognized as the golden standard for the quantification of viable bacteria. However, SPC usually takes one to several days to grow individual cells into a visible colony, which greatly hampers its application in rapid bacteria enumeration. Here we present a microdroplet turbidity imaging based digital standard plate count (dSPC) method to overcome this hurdle. Instead of cultivating on agar plates, bacteria are encapsulated in monodisperse microdroplets for single-cell cultivation. Proliferation of the encapsulated bacterial cell produced a detectable change in microdroplet turbidity, which allowed, after just a few bacterial doubling cycles (i.e., a few hours), enumeration of viable bacteria by visible-light imaging. Furthermore, a dSPC platform integrating a power-free droplet generator with smartphone-based turbidity imaging was established. As proof-of-concept demonstrations, a series of Gram-negative bacteria (Escherichia coli) and Gram-positive bacteria (Bacillus subtilis) samples were quantified via the smartphone dSPC accurately within 6 hours, representing a detection sensitivity of 100 CFU ml-1 and at least 3 times faster. In addition, Enterobacter sakazakii (E. sakazakii) in infant milk powder as a real sample was enumerated within 6 hours, in contrast to the 24 hours needed in traditional SPC. Results with high accuracy and reproducibility were achieved, with no difference in counts found between dSPC and SPC. By enabling label-free, rapid, portable and low-cost enumeration and cultivation of viable bacteria onsite, smartphone dSPC forms the basis for a temporally and geographically trackable network for surveying live microbes globally where every citizen with a cellphone can contribute anytime and anywhere.

Liming Poultry Manures to Kill Pathogens and Decrease Soluble Phosphorus

DOE Office of Scientific and Technical Information (OSTI.GOV)

Maguire,R.; Hesterberg, D.; Gernat, A.

2006-01-01

Received for publication September 9, 2005. Stabilizing phosphorus (P) in poultry waste to reduce P losses from manured soils is important to protect surface waters, while pathogens in manures are an emerging issue. This study was conducted to evaluate CaO and Ca(OH){sub 2} for killing manure bacterial populations (pathogens) and stabilizing P in poultry wastes and to investigate the influence on soils following amendment with the treated wastes. Layer manure and broiler litter varying in moisture content were treated with CaO and Ca(OH){sub 2} at rates of 2.5, 5, 10, and 15% by weight. All treated wastes were analyzed formore » microbial plate counts, pH, and water-soluble phosphorus (WSP), while a few selected layer manures were analyzed by phosphorus X-ray absorption near edge structure (XANES). A loamy sand and a silt loam were amended with broiler litter and layer manure treated with CaO at rates of 0, 2.5, 5, 10, and 15% and soil WSP and pH were measured at times 1, 8, and 29 d. Liming reduced bacterial populations, with greater rates of lime leading to greater reductions; for example 10% CaO applied to 20% solids broiler litter reduced the plate counts from 793 000 to 6500 mL{sup -1}. Liming also reduced the WSP in the manures by over 90% in all cases where at least 10% CaO was added. Liming the manures also reduced WSP in soils immediately following application and raised soil pH. The liming process used successfully reduced plate counts and concerns about P losses in runoff following land application of these limed products due to decreased WSP.« less

Liming poultry manures to decrease soluble phosphorus and suppress the bacteria population.

PubMed

Maguire, R O; Hesterberg, D; Gernat, A; Anderson, K; Wineland, M; Grimes, J

2006-01-01

Stabilizing phosphorus (P) in poultry waste to reduce P losses from manured soils is important to protect surface waters, while pathogens in manures are an emerging issue. This study was conducted to evaluate CaO and Ca(OH)2 for killing manure bacterial populations (pathogens) and stabilizing P in poultry wastes and to investigate the influence on soils following amendment with the treated wastes. Layer manure and broiler litter varying in moisture content were treated with CaO and Ca(OH)2 at rates of 2.5, 5, 10, and 15% by weight. All treated wastes were analyzed for microbial plate counts, pH, and water-soluble phosphorus (WSP), while a few selected layer manures were analyzed by phosphorus X-ray absorption near edge structure (XANES). A loamy sand and a silt loam were amended with broiler litter and layer manure treated with CaO at rates of 0, 2.5, 5, 10, and 15% and soil WSP and pH were measured at times 1, 8, and 29 d. Liming reduced bacterial populations, with greater rates of lime leading to greater reductions; for example 10% CaO applied to 20% solids broiler litter reduced the plate counts from 793,000 to 6500 mL-1. Liming also reduced the WSP in the manures by over 90% in all cases where at least 10% CaO was added. Liming the manures also reduced WSP in soils immediately following application and raised soil pH. The liming process used successfully reduced plate counts and concerns about P losses in runoff following land application of these limed products due to decreased WSP.

UV inactivation of pathogenic and indicator microorganisms

DOE Office of Scientific and Technical Information (OSTI.GOV)

Chang, J.C.; Ossoff, S.F.; Lobe, D.C.

1985-06-01

Survival was measured as a function of the dose of germicidal UV light for the bacteria Escherichia coli, Salmonella typhi, Shigella sonnei, Streptococcus faecalis, Staphylococcus aureus, and Bacillus subtilis spores, the enteric viruses poliovirus type 1 and simian rotavirus SA11, the cysts of the protozoan Acanthamoeba castellanii, as well as for total coliforms and standard plate count microorganisms from secondary effluent. The doses of UV light necessary for a 99.9% inactivation of the cultured vegetative bacteria, total coliforms, and standard plate count microorganisms were comparable. However, the viruses, the bacterial spores, and the amoebic cysts required about 3 to 4more » times, 9 times, and 15 times, respectively, the dose required for E. coli. These ratios covered a narrower relative dose range than that previously reported for chlorine disinfection of E. coli, viruses, spores, and cysts.« less

Bacteriological and genetic assessment of game meat from Japanese wild boars.

PubMed

Naya, Yuka; Horiuchi, Motohiro; Ishiguro, Naotaka; Shinagawa, Morikazu

2003-01-15

Bacterial tests were used to assess bacterial contamination of game meat from Japanese wild boars. The bacterial contamination of wild boar meat was less than that of domestic pork, as determined by aerobic plate counts (APC) and coliform counts. None of the meat examined in this study was contaminated by Salmonella or E. coli O-157. To detect adulteration by domestic pig meat or European wild boar meat, 46 samples of game meat sold as Japanese wild boar were examined genetically. A total of 17 samples showed genetic haplotypes of European and Asian domestic pigs in the D-loop of mitochondrial DNA (mtDNA), and 16 samples showed nuclear glucosephosphate isomerase-processed pseudogene (GPIP) genotypes of European domestic pigs. The European GPIP genotypes of these samples were confirmed by PCR-RFLP analysis. These results indicate that some game meat sold as Japanese wild boar is adulterated by cross-breeding between pigs and wild boars or by contamination with meat from domestic pigs or European wild boars.

Absolute Enumeration of Probiotic Strains Lactobacillus acidophilus NCFM® and Bifidobacterium animalis subsp. lactis Bl-04 ® via Chip-Based Digital PCR.

PubMed

Hansen, Sarah J Z; Morovic, Wesley; DeMeules, Martha; Stahl, Buffy; Sindelar, Connie W

2018-01-01

The current standard for enumeration of probiotics to obtain colony forming units by plate counts has several drawbacks: long time to results, high variability and the inability to discern between bacterial strains. Accurate probiotic cell counts are important to confirm the delivery of a clinically documented dose for its associated health benefits. A method is described using chip-based digital PCR (cdPCR) to enumerate Bifidobacterium animalis subsp. lactis Bl-04 and Lactobacillus acidophilus NCFM both as single strains and in combination. Primers and probes were designed to differentiate the target strains against other strains of the same species using known single copy, genetic differences. The assay was optimized to include propidium monoazide pre-treatment to prevent amplification of DNA associated with dead probiotic cells as well as liberation of DNA from cells with intact membranes using bead beating. The resulting assay was able to successfully enumerate each strain whether alone or in multiplex. The cdPCR method had a 4 and 5% relative standard deviation (RSD) for Bl-04 and NCFM, respectively, making it more precise than plate counts with an industry accepted RSD of 15%. cdPCR has the potential to replace traditional plate counts because of its precision, strain specificity and the ability to obtain results in a matter of hours.

Kynetic resazurin assay (KRA) for bacterial quantification of foodborne pathogens

NASA Astrophysics Data System (ADS)

Arenas, Yaxal; Mandel, Arkady; Lilge, Lothar

2012-03-01

Fast detection of bacterial concentrations is important for the food industry and for healthcare. Early detection of infections and appropriate treatment is essential since, the delay of treatments for bacterial infections tends to be associated with higher mortality rates. In the food industry and in healthcare, standard procedures require the count of colony-forming units in order to quantify bacterial concentrations, however, this method is time consuming and reports require three days to be completed. An alternative is metabolic-colorimetric assays which provide time efficient in vitro bacterial concentrations. A colorimetric assay based on Resazurin was developed as a time kinetic assay (KRA) suitable for bacterial concentration measurements. An optimization was performed by finding excitation and emission wavelengths for fluorescent acquisition. A comparison of two non-related bacteria, foodborne pathogens Escherichia coli and Listeria monocytogenes, was performed in 96 well plates. A metabolic and clonogenic dependence was established for fluorescent kinetic signals.

Infection control in digital intraoral radiography: evaluation of microbiological contamination of photostimulable phosphor plates in barrier envelopes.

PubMed

MacDonald, David S; Waterfield, J Douglas

2011-01-01

The detectors (both solid-state sensors and photostimulable phosphor [PSP] plates) used for digital intraoral radiography cannot be autoclaved, and barriers are typically used to prevent the spread of infection. The aim of this study was to determine the effectiveness of a barrier envelope system for PSP plates. Disinfected PSP plates were aseptically inserted into barrier envelopes and placed in a periapical location. One PSP plate was placed in each of 28 patients, and 12 plates in each of 2 volunteers (D.S.M., J.D.W.). After retrieval, each PSP plate was removed from its barrier envelope, immersed in trypticase soy broth and aliquots were plated on trypticase soy agar. Bacterial colonies were counted 2 days later. Fifty-two PSP plates in barrier envelopes were evaluated for contamination. Quality assurance of the PSP plates before clinical placement revealed defects in the integrity of 4 barrier envelopes, caused by forceps-related damage or failure to achieve a uniform seal. These defects allowed substantial contamination. Contamination also occurred as a result of failure to extract the PSP plate from the barrier envelope cleanly. Of the 44 barriers with no obvious defects that were placed by either final-year dental students or a radiologist, only 3 allowed bacterial contamination of the PSP plate. Detectors contained in barrier envelopes remain a potential source of contamination. PSP plates must be disinfected between removal from a contaminated barrier envelope and placement in a new barrier envelope. In addition, placement into the barrier envelope should ideally be carried out under aseptic conditions. Finally, the integrity of each sealed barrier envelope must be verified visually before release to the clinic.

High-Throughput Quantification of Bacterial-Cell Interactions Using Virtual Colony Counts

PubMed Central

Hoffmann, Stefanie; Walter, Steffi; Blume, Anne-Kathrin; Fuchs, Stephan; Schmidt, Christiane; Scholz, Annemarie; Gerlach, Roman G.

2018-01-01

The quantification of bacteria in cell culture infection models is of paramount importance for the characterization of host-pathogen interactions and pathogenicity factors involved. The standard to enumerate bacteria in these assays is plating of a dilution series on solid agar and counting of the resulting colony forming units (CFU). In contrast, the virtual colony count (VCC) method is a high-throughput compatible alternative with minimized manual input. Based on the recording of quantitative growth kinetics, VCC relates the time to reach a given absorbance threshold to the initial cell count using a series of calibration curves. Here, we adapted the VCC method using the model organism Salmonella enterica sv. Typhimurium (S. Typhimurium) in combination with established cell culture-based infection models. For HeLa infections, a direct side-by-side comparison showed a good correlation of VCC with CFU counting after plating. For MDCK cells and RAW macrophages we found that VCC reproduced the expected phenotypes of different S. Typhimurium mutants. Furthermore, we demonstrated the use of VCC to test the inhibition of Salmonella invasion by the probiotic E. coli strain Nissle 1917. Taken together, VCC provides a flexible, label-free, automation-compatible methodology to quantify bacteria in in vitro infection assays. PMID:29497603

LENGTH-HETEROGENEITY POLYMERASE CHAIN REACTION (LH-PCR) AS AN INDICATOR OF STREAM SANITARY AND ECOLOGICAL CONDITION: OPTIMAL SAMPLE SIZE AND HOLDING CONDITIONS

EPA Science Inventory

The use of coliform plate count data to assess stream sanitary and ecological condition is limited by the need to store samples at 4oC and analyze them within a 24-hour period. We are testing LH-PCR as an alternative tool to assess the bacterial load of streams, offering a cost ...

Optimization of single plate-serial dilution spotting (SP-SDS) with sample anchoring as an assured method for bacterial and yeast cfu enumeration and single colony isolation from diverse samples.

PubMed

Thomas, Pious; Sekhar, Aparna C; Upreti, Reshmi; Mujawar, Mohammad M; Pasha, Sadiq S

2015-12-01

We propose a simple technique for bacterial and yeast cfu estimations from diverse samples with no prior idea of viable counts, designated as single plate-serial dilution spotting (SP-SDS) with the prime recommendation of sample anchoring (10 0 stocks). For pure cultures, serial dilutions were prepared from 0.1 OD (10 0 ) stock and 20 μl aliquots of six dilutions (10 1 -10 6 ) were applied as 10-15 micro-drops in six sectors over agar-gelled medium in 9-cm plates. For liquid samples 10 0 -10 5 dilutions, and for colloidal suspensions and solid samples (10% w/v), 10 1 -10 6 dilutions were used. Following incubation, at least one dilution level yielded 6-60 cfu per sector comparable to the standard method involving 100 μl samples. Tested on diverse bacteria, composite samples and Saccharomyces cerevisiae , SP-SDS offered wider applicability over alternative methods like drop-plating and track-dilution for cfu estimation, single colony isolation and culture purity testing, particularly suiting low resource settings.

The association between bedding material and the bacterial counts of Staphylococcus aureus, Streptococcus uberis and coliform bacteria on teat skin and in teat canals in lactating dairy cattle.

PubMed

Paduch, Jan-Hendrik; Mohr, Elmar; Krömker, Volker

2013-05-01

Several mastitis-causing pathogens are able to colonize the bovine teat canal. The objective of this study was to investigate the association between the treatment of sawdust bedding with a commercial alkaline conditioner and the bacterial counts on teat skin and in the teat canal. The study used a crossover design. Ten lactating Holstein cows that were free of udder infections and mastitis were included in the study. The animals were bedded on either untreated sawdust or sawdust that had been treated with a hydrated lime-based conditioner. Once a day, fresh bedding material was added. After 3 weeks, the bedding material was removed from the cubicles, fresh bedding material was provided, and the cows were rotated between the two bedding material groups. Teat skin and teat canals were sampled using the wet and dry swab technique after weeks 1, 2, 3, 4, 5 and 6. Staphylococcus aureus, Streptococcus uberis, Escherichia coli and other coliform bacteria were detected in the resulting agar plate cultures. The treatment of the bedding material was associated with the teat skin bacterial counts of Str. uberis, Esch. coli and other coliform bacteria. An association was also found between the bedding material and the teat canal bacterial counts of coliform bacteria other than Esch. coli. For Staph. aureus, no associations with the bedding material were found. In general, the addition of a hydrated lime-based conditioner to sawdust reduces the population sizes of environmental pathogens on teat skin and in teat canals.

Intestinal manipulation affects mucosal antimicrobial defense in a mouse model of postoperative ileus

PubMed Central

Hieggelke, Lena; Schneiker, Bianca; Lysson, Mariola; Stoffels, Burkhard; Nuding, Sabine; Wehkamp, Jan; Kikhney, Judith; Moter, Annette; Kalff, Joerg C.

2018-01-01

Aim To explore the effects of abdominal surgery and interleukin-1 signaling on antimicrobial defense in a model of postoperative ileus. Methods C57BL/6 and Interleukin-1 receptor type I (IL-1R1) deficient mice underwent intestinal manipulation to induce POI. Expression of mucosal IL-1α, IL-1β and IL-1R1 and several antimicrobial peptides and enzymes were measured by quantitative PCR or ELISA, western blotting or immunohistochemistry. Bacterial overgrowth was determined by fluorescent in-situ hybridization and counting of jejunal luminal bacteria. Translocation of aerobic and anaerobic bacteria into the intestinal wall, mesenteric lymph nodes, liver and spleen was determined by counting bacterial colonies on agar plates 48h after plating of tissue homogenates. Antimicrobial activity against E. coli and B. vulgatus was analyzed in total and cationic fractions of small bowel mucosal tissue homogenates by a flow cytometry-based bacterial depolarization assay. Results Jejunal bacterial overgrowth was detected 24h after surgery. At the same time point, but not in the early phase 3h after surgery, bacterial translocation into the liver and mesenteric lymph nodes was observed. Increased antimicrobial activity against E. coli was induced within early phase of POI. Basal antimicrobial peptide and enzyme gene expression was higher in the ileal compared to the jejunal mucosa. The expression of lysozyme 1, cryptdin 1, cryptdin 4 and mucin 2 were reduced 24h after surgery in the ileal mucosa and mucin 2 was also reduced in the jejunum. Postoperative IL-1α and IL-1β were increased in the postoperative mucosa. Deficiency of IL-1R1 affected the expression of antimicrobial peptides during homeostasis and POI. Conclusion Small bowel antimicrobial capacity is disturbed during POI which is accompanied by bacterial overgrowth and translocation. IL-1R1 is partially involved in the gene expression of mucosal antimicrobial peptides. Altered small bowel antimicrobial activity may contribute also to POI development and manifestation in patients undergoing abdominal surgery. PMID:29652914

Protocol for evaluating the efficacy of cetylpyridinium chloride as a beef hide intervention.

PubMed

Bosilevac, Joseph M; Wheeler, Tommy L; Rivera-Betancourt, Mildred; Nou, Xiangwu; Arthur, Terrance M; Shackelford, Steven D; Kent, Matthew P; Jaroni, Divya; Osborn, Matthew S; Rossman, Michelle; Reagan, James O; Koohmaraie, Mohammad

2004-02-01

The objective of this study was to establish the necessary protocols and assess the efficacy of cetylpyridinium chloride (CPC) as an antimicrobial intervention on beef cattle hides. Experiments using CPC were conducted to determine (i) the methods of neutralization needed to obtain valid efficacy measurements, (ii) the effect of concentration and dwell time after treatment, (iii) the effect of CPC on hide and carcass microbial populations when cattle were treated at a feedlot and then transported to a processing facility for harvest, and (iv) the effectiveness of spray pressure and two-spray combinations of CPC and water to reduce hide microbial populations. Residual CPC in hide sponge samples prevented bacterial growth. Dey-Engley neutralization media at 7.8% and a centrifugation step were necessary to overcome this problem. All dwell times, ranging from 30 s to 4 h, after 1% CPC application to cattle hides resulted in aerobic plate counts and Enterobacteriaceae counts 1.5 log CFU/100 cm2 lower than controls. The most effective dose of CPC was 1%, which reduced aerobic plate counts and Enterobacteriaceae counts 2 and 1 log CFU/100 cm2, respectively. Low-pressure application of 1% CPC at the feedlot, transport to the processing facility, and harvest within 5 h of application resulted in no effect on Escherichia coli O157 prevalence on hides or preevisceration carcasses. Two high-pressure CPC washes lowered aerobic plate counts and Enterobacteriaceae counts by 4 log CFU/100 cm2, and two medium-pressure CPC washes were only slightly less effective. These results indicate that under the proper conditions, CPC may still be effective for reducing microbial populations on cattle hides. Further study is warranted to determine if this effect will result in reduction of hide-to-carcass contamination during processing.

Is hand hygiene before putting on nonsterile gloves in the intensive care unit a waste of health care worker time?--a randomized controlled trial.

PubMed

Rock, Clare; Harris, Anthony D; Reich, Nicholas G; Johnson, J Kristie; Thom, Kerri A

2013-11-01

Hand hygiene (HH) is recognized as a basic effective measure in prevention of nosocomial infections. However, the importance of HH before donning nonsterile gloves is unknown, and few published studies address this issue. Despite the lack of evidence, the World Health Organization and other leading bodies recommend this practice. The aim of this study was to assess the utility of HH before donning nonsterile gloves prior to patient contact. A prospective, randomized, controlled trial of health care workers entering Contact Isolation rooms in intensive care units was performed. Baseline finger and palm prints were made from dominant hands onto agar plates. Health care workers were then randomized to directly don nonsterile gloves or perform HH and then don nonsterile gloves. Postgloving finger and palm prints were then made from the gloved hands. Plates were incubated and colony-forming units (CFU) of bacteria were counted. Total bacterial colony counts of gloved hands did not differ between the 2 groups (6.9 vs 8.1 CFU, respectively, P = .52). Staphylococcus aureus was identified from gloves (once in "hand hygiene prior to gloving" group, twice in "direct gloving" group). All other organisms were expected commensal flora. HH before donning nonsterile gloves does not decrease already low bacterial counts on gloves. The utility of HH before donning nonsterile gloves may be unnecessary. Copyright © 2013 Association for Professionals in Infection Control and Epidemiology, Inc. Published by Mosby, Inc. All rights reserved.

Microbiological quality and potential public health risks of export meat from springbok (Antidorcas marsupialis) in Namibia.

PubMed

Magwedere, K; Shilangale, R; Mbulu, R S; Hemberger, Y; Hoffman, L C; Dziva, F

2013-01-01

To assess the microbiological quality and safety of export game meat; i) a total of 80 pooled meat samples for aerobic plate count (APC) and Enterobacteriaceae ii) water used in harvesting and processing for microbiological quality and iii) meat and rectal contents for Salmonella spp. and Shiga toxin Escherichia coli (STEC) were evaluated in 2009 and 2010. No differences (p>0.05) in the APCs were observed between the years, but the mean Enterobacteriaceae count for 2009 was 1.33 ± 0.69 log(10)cfu/cm(2) compared to 2.93 ± 1.50 log(10)cfu/cm(2) for 2010. Insignificant Heterotrophic Plate Count (HPC) levels were detected in 9/23 field water samples, while fecal bacterial (coliforms, Clostridium perfringens and enterococci) were absent in all samples. No Salmonella spp. was isolated and all E. coli isolates from meat were negative for STEC virulence genes (stx1, stx2, eae and hlyA), suggesting a negligible role by springbok in the epidemiology of STEC and Salmonella. Copyright © 2012 Elsevier Ltd. All rights reserved.

Proteus mirabilis viability after lithotripsy of struvite calculi

NASA Astrophysics Data System (ADS)

Prabakharan, Sabitha; Teichman, Joel M. H.; Spore, Scott S.; Sabanegh, Edmund; Glickman, Randolph D.; McLean, Robert J. C.

2000-05-01

Urinary calculi composed of struvite harbor urease-producing bacteria within the stone. The photothermal mechanism of holmium:YAG lithotripsy is uniquely different than other lithotripsy devices. We postulated that bacterial viability of struvite calculi would be less for calculi fragmented with holmium:YAG irradiation compared to other lithotripsy devices. Human calculi of known struvite composition (greater than 90% magnesium ammonium phosphate hexahydrate) were incubated with Proteus mirabilis. Calculi were fragmented with no lithotripsy (controls), or shock wave, intracorporeal ultrasonic, electrohydraulic, pneumatic, holmium:YAG or pulsed dye laser lithotripsy. After lithotripsy, stone fragments were sonicated and specimens were serially plated for 48 hours at 38 C. Bacterial counts and the rate of bacterial sterilization were compared. Median bacterial counts (colony forming units per ml) were 8 X 106 in controls and 3 X 106 in shock wave, 3 X 107 in ultrasonic, 4 X 105 in electrohydraulic, 8 X 106 in pneumatic, 5 X 104 in holmium:YAG and 1 X 106 in pulsed dye laser lithotripsy, p less than 0.001. The rate of bacterial sterilization was 50% for holmium:YAG lithotripsy treated stones versus 0% for each of the other cohorts, p less than 0.01. P. mirabilis viability is less after holmium:YAG irradiation compared to other lithotripsy devices.

Results from raw milk microbiological tests do not predict the shelf-life performance of commercially pasteurized fluid milk.

PubMed

Martin, N H; Ranieri, M L; Murphy, S C; Ralyea, R D; Wiedmann, M; Boor, K J

2011-03-01

Analytical tools that accurately predict the performance of raw milk following its manufacture into commercial food products are of economic interest to the dairy industry. To evaluate the ability of currently applied raw milk microbiological tests to predict the quality of commercially pasteurized fluid milk products, samples of raw milk and 2% fat pasteurized milk were obtained from 4 New York State fluid milk processors for a 1-yr period. Raw milk samples were examined using a variety of tests commonly applied to raw milk, including somatic cell count, standard plate count, psychrotrophic bacteria count, ropy milk test, coliform count, preliminary incubation count, laboratory pasteurization count, and spore pasteurization count. Differential and selective media were used to identify groups of bacteria present in raw milk. Pasteurized milk samples were held at 6°C for 21 d and evaluated for standard plate count, coliform count, and sensory quality throughout shelf-life. Bacterial isolates from select raw and pasteurized milk tests were identified using 16S ribosomal DNA sequencing. Linear regression analysis of raw milk test results versus results reflecting pasteurized milk quality consistently showed low R(2) values (<0.45); the majority of R(2) values were <0.25, indicating small relationship between the results from the raw milk tests and results from tests used to evaluate pasteurized milk quality. Our findings suggest the need for new raw milk tests that measure the specific biological barriers that limit shelf-life and quality of fluid milk products. Copyright © 2011 American Dairy Science Association. Published by Elsevier Inc. All rights reserved.

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

On-farm and postharvest processing sources of bacterial contamination to melon rinds.

PubMed

Gagliardi, J V; Millner, P D; Lester, G; Ingram, D

2003-01-01

Multistate and international foodborne illness outbreaks, particularly involving cantaloupe and often involving rare Salmonella spp., have increased dramatically over the past 13 years. This study assessed the sources and extent of melon rind contamination in production fields and at processing and packing facilities. In the spring of 1999, cantaloupe (Cucumis melo L. [reticulatus group] cv. Cruiser) sampled from two sites in the Rio Grande River Valley showed that postharvest-processed melon rinds often had greater plate counts of bacterial contaminants than field-fresh melons. Cantaloupe in the field had 2.5 to 3.5 log CFU g(-1) rind total coliforms by aerobic plate counts, whereas washed melons had 4.0 to 5.0 log CFU g(-1). In the fall of 1999, coliforms on honeydew melons (C. melo [inodorous group] cv. Honey Brew) ranged from 2.6 to 3.7 log CFU g(-1) after processing, and total and fecal coliforms and enterococci never fell below 2.5 log CFU g(-1). A hydrocooler at another site contaminated cantaloupe rinds with up to 3.4 log CFU g(-1) total and fecal enterococci; a secondary rinse with chlorinated water incompletely removed these bacteria. Sources of coliforms and enterococci were at high levels in melon production soils, especially in furrows that were flood irrigated, in standing water at one field, and in irrigation water at both sites. At one processing facility, wash water pumped from the Rio Grande River may not have been sufficiently disinfected prior to use. Because soil, irrigation water, and process water were potential sources of bacterial contamination, monitoring and management on-farm and at processing and packing facilities should focus on water quality as an important control point for growers and packers to reduce bacterial contamination on melon rinds.

Comparison of Standard Culture-Based Method to Culture-Independent Method for Evaluation of Hygiene Effects on the Hand Microbiome

PubMed Central

Leff, J.; Henley, J.; Tittl, J.; De Nardo, E.; Butler, M.; Griggs, R.; Fierer, N.

2017-01-01

ABSTRACT Hands play a critical role in the transmission of microbiota on one’s own body, between individuals, and on environmental surfaces. Effectively measuring the composition of the hand microbiome is important to hand hygiene science, which has implications for human health. Hand hygiene products are evaluated using standard culture-based methods, but standard test methods for culture-independent microbiome characterization are lacking. We sampled the hands of 50 participants using swab-based and glove-based methods prior to and following four hand hygiene treatments (using a nonantimicrobial hand wash, alcohol-based hand sanitizer [ABHS], a 70% ethanol solution, or tap water). We compared results among culture plate counts, 16S rRNA gene sequencing of DNA extracted directly from hands, and sequencing of DNA extracted from culture plates. Glove-based sampling yielded higher numbers of unique operational taxonomic units (OTUs) but had less diversity in bacterial community composition than swab-based sampling. We detected treatment-induced changes in diversity only by using swab-based samples (P < 0.001); we were unable to detect changes with glove-based samples. Bacterial cell counts significantly decreased with use of the ABHS (P < 0.05) and ethanol control (P < 0.05). Skin hydration at baseline correlated with bacterial abundances, bacterial community composition, pH, and redness across subjects. The importance of the method choice was substantial. These findings are important to ensure improvement of hand hygiene industry methods and for future hand microbiome studies. On the basis of our results and previously published studies, we propose recommendations for best practices in hand microbiome research. PMID:28351915

The presence of Enterococcus, coliforms and E. coli in a commercial yeast manufacturing process.

PubMed

O'Brien, S S; Lindsay, D; von Holy, A

2004-07-01

This study evaluated a typical commercial yeast manufacturing process for bacterial contamination. Product line samples of a commercial yeast manufacturing process and the corresponding seed yeast manufacturing process were obtained upstream from the final compressed and dry yeast products. All samples were analysed before (non-PI) and after preliminary incubation (PI) at 37 degrees C for 24 h. The PI procedure was incorporated for amplification of bacterial counts below the lower detection limit. Enterococcus, coliform and Escherichia coli counts were quantified by standard pour-plate techniques using selective media. Presence at all stages and progressive increases in counts of Enterococcus, coliforms and E. coli during processing in the commercial manufacturing operation suggested that the primary source of contamination of both compressed and dry yeast with these bacteria was the seed yeast manufacturing process and that contamination was amplified throughout the commercial yeast manufacturing process. This was confirmed by surveys of the seed yeast manufacturing process which indicated that contamination of the seed yeast with Enterococcus, coliforms and E. coli occurred during scale up of seed yeast biomass destined as inoculum for the commercial fermentation.

Hydrodebridement of wounds: effectiveness in reducing wound bacterial contamination and potential for air bacterial contamination.

PubMed

Bowling, Frank L; Stickings, Daryl S; Edwards-Jones, Valerie; Armstrong, David G; Boulton, Andrew Jm

2009-05-08

The purpose of this study was to assess the level of air contamination with bacteria after surgical hydrodebridement and to determine the effectiveness of hydro surgery on bacterial reduction of a simulated infected wound. Four porcine samples were scored then infected with a broth culture containing a variety of organisms and incubated at 37 degrees C for 24 hours. The infected samples were then debrided with the hydro surgery tool (Versajet, Smith and Nephew, Largo, Florida, USA). Samples were taken for microbiology, histology and scanning electron microscopy pre-infection, post infection and post debridement. Air bacterial contamination was evaluated before, during and after debridement by using active and passive methods; for active sampling the SAS-Super 90 air sampler was used, for passive sampling settle plates were located at set distances around the clinic room. There was no statistically significant reduction in bacterial contamination of the porcine samples post hydrodebridement. Analysis of the passive sampling showed a significant (p < 0.001) increase in microbial counts post hydrodebridement. Levels ranging from 950 colony forming units per meter cubed (CFUs/m3) to 16780 CFUs/m3 were observed with active sampling of the air whilst using hydro surgery equipment compared with a basal count of 582 CFUs/m3. During removal of the wound dressing, a significant increase was observed relative to basal counts (p < 0.05). Microbial load of the air samples was still significantly raised 1 hour post-therapy. The results suggest a significant increase in bacterial air contamination both by active sampling and passive sampling. We believe that action might be taken to mitigate fallout in the settings in which this technique is used.

Characterization of the dominant bacterial communities during storage of Norway lobster and Norway lobster tails (Nephrops norvegicus) based on 16S rDNA analysis by PCR-DGGE.

PubMed

Bekaert, Karen; Devriese, Lisa; Maes, Sara; Robbens, Johan

2015-04-01

The aim of this study was to investigate the microbial quality of whole Norway lobster (Nephrops norvegicus) and Norway lobster tails to optimize handling conditions. This was done by assessing the total viable count (TVC) and characterizing the dominant microbiota. The cultivable microorganisms were quantified via classical microbiological plating methods. To characterize as many bacterial species present as possible, we performed advanced molecular identification techniques (PCR-DGGE). The initial TVC of fresh Norway lobster meat was high (3.0 log cfu/g) as compared to fish. No significant difference between whole Norway lobster and Norway lobster tails could be found during the storage period. From day 6 of storage, a significant difference between Plate Count Agar (PCA) and Marine Agar (MA) was observed. The microbiota of Norway lobster was dominated by members of the Gram-negative genera such as Psychrobacter spp., Pseudoalteromonas spp., Pseudomonas spp., Luteimonas spp., and Aliivibrio spp. From these bacteria, mainly Psychrobacter spp. and Pseudomonas spp. remained present until the end of the storage period. These are known spoilage organisms in fishery products. Other known spoilage organisms of crustaceans such as Photobacterium spp. could not be identified. Copyright © 2014 Elsevier Ltd. All rights reserved.

A Randomized Clinical Trial Evaluating the Effects of Oligosaccharides on Transfer of Passive Immunity in Neonatal Dairy Calves.

PubMed

Short, D M; Moore, D A; Sischo, W M

2016-07-01

Bacterial contamination of colostrum is common and can decrease IgG absorption in neonatal calves. Strategies that mitigate this situation without complicating colostrum management will benefit dairy calf health and survival. To evaluate the effects of supplementing colostrum with oligosaccharides (OS) on serum IgG concentration and apparent efficiency of absorption of IgG (AEA%) in calves fed unpasteurized colostrum and characterize these outcomes with respect to colostrum bacterial exposures. One hundred twenty-three neonatal dairy calves. Randomized, blinded, controlled clinical trial conducted at a commercial dairy operation. Calves were enrolled at birth in 1 of 4 treatment groups. Data were complete for 123 calves, which were distributed across the treatment groups as follows: mannan-oligosaccharides (MOS), n = 33; Saccharomyces galacto-oligosaccharides (SGOS), n = 31; Bifidobacterium galacto-oligosaccharides (BGOS), n = 28; and lactose control (CON), n = 31. A commercial radial immunodiffusion kit was used to determine colostrum and serum IgG concentrations. Conventional microbiology methods were used to enumerate colostrum bacterial counts. Bacterial counts were not significantly different among treatment groups. Total bacterial plate counts (TPC) were relatively low for the majority of colostrum samples, but TPC had a significant negative effect on serum IgG concentration and AEA% in the lactose-supplemented control group but not the OS treatment groups. These results suggest that a complement of OS structures may mitigate adverse effects of bacteria on transfer of passive immunity (TPI). Copyright © 2016 The Authors. Journal of Veterinary Internal Medicine published by Wiley Periodicals, Inc. on behalf of the American College of Veterinary Internal Medicine.

Enumeration of the contaminating bacterial microbiota in unfermented pasteurized milks enriched with probiotic bacteria.

PubMed

Champagne, C P; Raymond, Y; Gonthier, J; Audet, P

2009-04-01

Pasteurized and unfermented milks supplemented with probiotic bacteria are appearing on the market. It then becomes a challenge to ascertain the undesirable contamination microbiota in the presence of a largely superior population of probiotic bacteria. A method to enumerate the contaminating microbial microbiota in such probiotic-enriched milks was developed. The probiotic cultures, Lactobacillus rhamnosus Lb-Immuni-T and Bifidobacterium animalis subsp. lactis BB-12(R), were added to a pasteurized unfermented milk to reach a minimum of 1 billion CFU per 250 mL portion, as ascertained by plating on de Man - Rogosa - Sharpe (MRS) agar in anaerobic conditions. No growth of B. animalis subsp. lactis BB-12 was noted on plate count agar (PCA) or Petrifilm plates, and the presence of this culture did not affect standard plate counts (SPC) of contaminating bacteria. However, L. rhamnosus formed colonies on PCA and Petrifilm plates. Attempts were thus made to inhibit the growth of the probiotic lactobacilli in PCA. The addition of 2% sodium phosphate (SP) or 5% glycerophosphate (GP) inhibited the growth of the lactobacilli in broths, but pin-point colonies of L. rhamnosus Lb-Immuni-T nevertheless appeared on PCA supplemented with phosphates. SPC could be obtained on PCA + 2% SP by only counting the large colonies, but this resulted in a significant (4.4 fold) underestimation of SPC values. On Petrifilm AC, at dilutions 0 to 2, all colonies were considered as being contaminants, while at dilutions 3 and 4, only large colonies were counted for SPC determinations. There was a direct correlation (R2 = 0.99) between SPC values with Petrifilm in uninoculated milks and those obtained on probiotic-enriched milks. The high correlation obtained over the 102 to 106 CFU/mL range of SPC values show that this Petrifilm method is appropriate to evaluate the microbiological quality of pasteurized milks enriched with L. rhamnosus Lb-Immuni-T and B. animalis subsp. lactis BB-12.

Microbial contamination of mobile phones in a health care setting in Alexandria, Egypt.

PubMed

Selim, Heba Sayed; Abaza, Amani Farouk

2015-01-01

This study aimed at investigating the microbial contamination of mobile phones in a hospital setting. Swab samples were collected from 40 mobile phones of patients and health care workers at the Alexandria University Students' Hospital. They were tested for their bacterial contamination at the microbiology laboratory of the High Institute of Public Health. Quantification of bacteria was performed using both surface spread and pour plate methods. Isolated bacterial agents were identified using standard microbiological methods. Methicillin-resistant Staphylococcus aureus was identified by disk diffusion method described by Bauer and Kirby. Isolated Gram-negative bacilli were tested for being extended spectrum beta lactamase producers using the double disk diffusion method according to the Clinical and Laboratory Standards Institute recommendations. All of the tested mobile phones (100%) were contaminated with either single or mixed bacterial agents. The most prevalent bacterial contaminants were methicillin-resistant S. aureus and coagulase-negative staphylococci representing 53% and 50%, respectively. The mean bacterial count was 357 CFU/ml, while the median was 13 CFU/ml using the pour plate method. The corresponding figures were 2,192 and 1,720 organisms/phone using the surface spread method. Mobile phones usage in hospital settings poses a risk of transmission of a variety of bacterial agents including multidrug-resistant pathogens as methicillin-resistant S. aureus. The surface spread method is an easy and useful tool for detection and estimation of bacterial contamination of mobile phones.

Microbial contamination of mobile phones in a health care setting in Alexandria, Egypt

PubMed Central

Selim, Heba Sayed; Abaza, Amani Farouk

2015-01-01

Aim: This study aimed at investigating the microbial contamination of mobile phones in a hospital setting. Methods: Swab samples were collected from 40 mobile phones of patients and health care workers at the Alexandria University Students’ Hospital. They were tested for their bacterial contamination at the microbiology laboratory of the High Institute of Public Health. Quantification of bacteria was performed using both surface spread and pour plate methods. Isolated bacterial agents were identified using standard microbiological methods. Methicillin-resistant Staphylococcus aureus was identified by disk diffusion method described by Bauer and Kirby. Isolated Gram-negative bacilli were tested for being extended spectrum beta lactamase producers using the double disk diffusion method according to the Clinical and Laboratory Standards Institute recommendations. Results: All of the tested mobile phones (100%) were contaminated with either single or mixed bacterial agents. The most prevalent bacterial contaminants were methicillin-resistant S. aureus and coagulase-negative staphylococci representing 53% and 50%, respectively. The mean bacterial count was 357 CFU/ml, while the median was 13 CFU/ml using the pour plate method. The corresponding figures were 2,192 and 1,720 organisms/phone using the surface spread method. Conclusions: Mobile phones usage in hospital settings poses a risk of transmission of a variety of bacterial agents including multidrug-resistant pathogens as methicillin-resistant S. aureus. The surface spread method is an easy and useful tool for detection and estimation of bacterial contamination of mobile phones. PMID:25699226

Evaluation of the methods for enumerating coliform bacteria from water samples using precise reference standards.

PubMed

Wohlsen, T; Bates, J; Vesey, G; Robinson, W A; Katouli, M

2006-04-01

To use BioBall cultures as a precise reference standard to evaluate methods for enumeration of Escherichia coli and other coliform bacteria in water samples. Eight methods were evaluated including membrane filtration, standard plate count (pour and spread plate methods), defined substrate technology methods (Colilert and Colisure), the most probable number method and the Petrifilm disposable plate method. Escherichia coli and Enterobacter aerogenes BioBall cultures containing 30 organisms each were used. All tests were performed using 10 replicates. The mean recovery of both bacteria varied with the different methods employed. The best and most consistent results were obtained with Petrifilm and the pour plate method. Other methods either yielded a low recovery or showed significantly high variability between replicates. The BioBall is a very suitable quality control tool for evaluating the efficiency of methods for bacterial enumeration in water samples.

Physiological responses of bacteria in biofilms to disinfection.

PubMed Central

Yu, F P; McFeters, G A

1994-01-01

In situ enumeration methods using fluorescent probes and a radioisotope labelling technique were applied to evaluate physiological changes of Klebsiella pneumoniae within biofilms after disinfection treatment. Chlorine (0.25 mg of free chlorine per liter [pH 7.2]) and monochloramine (1 mg/liter [pH 9.0]) were employed as disinfectants in the study. Two fluorgenic compounds, 5-cyano-2,3-ditolyl tetrazolium chloride and rhodamine 123, and tritiated uridine incorporation were chosen for assessment of physiological activities. Results obtained by these methods were compared with those from the plate count and direct viable count methods. 5-Cyano-2,3-ditolyl tetrazolium chloride is an indicator of bacterial respiratory activity, rhodamine 123 is incorporated into bacteria in response to transmembrane potential, and the incorporation of uridine represents the global RNA turnover rate. The results acquired by these methods following disinfection exposure showed a range of responses and suggested different physiological reactions in biofilms exposed to chlorine and monochloramine. The direct viable count response and respiratory activity were affected more by disinfection than were the transmembrane potential and RNA turnover rate on the basis of comparable efficiency as evaluated by plate count enumeration. Information revealed by these approaches can provide different physiological insights that may be used in evaluating the efficacy of biofilm disinfection. PMID:8074525

Bacterial contamination of the hands of food handlers as indicator of hand washing efficacy in some convenient food industries in South Africa.

PubMed

Aa, Lambrechts; Is, Human; Jh, Doughari; Jfr, Lues

2014-07-01

Hands of ready-to-eat food service employees have been shown to be vectors in the spread of foodborne disease, mainly because of poor personal hygiene and accounting for approximately 97% of food borne illnesses in food service establishments and homes. Our objective was to evaluate the efficacy of hand washing practices and sanitation before commencing work among food handlers in the convenient food industry in Gauteng, South Africa. A total of 230 samples were collected, involving 100% of the food handlers, in 8 selected convenient food outlets with their main focus on preparing ready-to-eat foods. The workers' cleaned and disinfected dominant hands were sampled for Total Plate Count (TPC), Staphylococcus aureus and Escherichia coli. Bacteria were isolated and counted using standard methods. The highest bacterial count from the hand samples was 7.4 x 10(3) cfu.cm(-2) and the lowest showed no detectable growth. Although hands with a count of 0 cfu.cm(-2) were found in all of the plants, the results indicated that all the plants exceeded the legal limit for food surfaces or hands of < 100 cfu.cm(-2) when the average bacterial counts on hands were compared. Sixty percent of the TPC analysed exceeded the legal limit and only 18% of the food handlers had no bacteria detectable on their hands. One sample tested positive for E. coli and S. aureus could not be detected on the hands of any of the food handlers. The study revealed that hand hygiene is unsatisfactory and may have serious implications for public health due to contamination of food from food handlers' hands. This therefore underlined the importance of further training to improve food handlers' knowledge of good hand washing practices.

Heat resistance of viable but non-culturable Escherichia coli cells determined by differential scanning calorimetry.

PubMed

Castro-Rosas, Javier; Gómez-Aldapa, Carlos Alberto; Villagómez Ibarra, José Roberto; Santos-López, Eva María; Rangel-Vargas, Esmeralda

2017-10-16

Several reports have suggested that the viable but non-culturable (VBNC) state is a resistant form of bacterial cells that allows them to remain in a dormant form in the environment. Nevertheless, studies on the resistance of VBNC bacterial cells to ecological factors are limited, mainly because techniques that allow this type of evaluation are lacking. Differential scanning calorimetry (DSC) has been used to study the thermal resistance of culturable bacteria but has never been used to study VBNC cells. In this work, the heat resistance of Escherichia coli cells in the VBNC state was studied using the DSC technique. The VBNC state was induced in E. coli ATCC 25922 by suspending bacterial cells in artificial sea water, followed by storage at 3 ± 2°C for 110 days. Periodically, the behaviour of E. coli cells was monitored by plate counts, direct viable counts and DSC. The entire bacterial population entered the VBNC state after 110 days of storage. The results obtained with DSC suggest that the VBNC state does not confer thermal resistance to E. coli cells in the temperature range analysed here. © FEMS 2017. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com.

The Rebirth of Culture in Microbiology through the Example of Culturomics To Study Human Gut Microbiota

PubMed Central

Lagier, Jean-Christophe; Hugon, Perrine; Khelaifia, Saber; Fournier, Pierre-Edouard; La Scola, Bernard

2015-01-01

SUMMARY Bacterial culture was the first method used to describe the human microbiota, but this method is considered outdated by many researchers. Metagenomics studies have since been applied to clinical microbiology; however, a “dark matter” of prokaryotes, which corresponds to a hole in our knowledge and includes minority bacterial populations, is not elucidated by these studies. By replicating the natural environment, environmental microbiologists were the first to reduce the “great plate count anomaly,” which corresponds to the difference between microscopic and culture counts. The revolution in bacterial identification also allowed rapid progress. 16S rRNA bacterial identification allowed the accurate identification of new species. Mass spectrometry allowed the high-throughput identification of rare species and the detection of new species. By using these methods and by increasing the number of culture conditions, culturomics allowed the extension of the known human gut repertoire to levels equivalent to those of pyrosequencing. Finally, taxonogenomics strategies became an emerging method for describing new species, associating the genome sequence of the bacteria systematically. We provide a comprehensive review on these topics, demonstrating that both empirical and hypothesis-driven approaches will enable a rapid increase in the identification of the human prokaryote repertoire. PMID:25567229

Prevention of Bacterial Biofilms Formation on Urinary Catheter by Selected Plant Extracts.

PubMed

Adesina, T D; Nwinyi, O C; Olugbuyiro, J A O

2015-02-01

In this study, we investigated the feasibility of using Psidium guajava, Mangifera indica and Ocimum gratissimum leaf extracts in preventing Escherichia coli biofilm formation. The plants extractions were done with methanol under cold extraction. The various concentrations 5.0, 10.0 and 20.0 mg mL(-1) were used to coat 63 catheters under mild heat from water bath. Biofilm formation on the catheter was induced using cultures of E. coli. Biofilm formation was evaluated using aerobic plate count and turbidity at 600 nm. From the obtained results, Psidium guajava, Mangifera indica and Ocimum gratissimum delayed the onset of biofilm formation for a week. Ocimum gratissimum coated catheter had the highest inhibitory effect at 5.0, 10.0 and 20.0 mg mL(-1) with bacterial count ranging from 2.2 x 10(5)-7.0 x 10(4) and 5.7 x 10(5)-3.7 x10(5) for 120 and 128 h, respectively. The Psidium guajava coated catheter had the lowest inhibitory effect at 5.0, 10.0 and 20.0 mg mL(-1), with bacterial count ranging between 4.3 x 10(5)-1.9 x 10(3) and 7.7 x 10(5)-3.8 x 10(5) for 120 and 128 h, respectively. Despite the antimicrobial activities, the differences in the activity of these plant extracts were statistically not significant (p < 0.05).

Selective cultivation and rapid detection of Staphylococcus aureus by computer vision.

PubMed

Wang, Yong; Yin, Yongguang; Zhang, Chaonan

2014-03-01

In this paper, we developed a selective growth medium and a more rapid detection method based on computer vision for selective isolation and identification of Staphylococcus aureus from foods. The selective medium consisted of tryptic soy broth basal medium, 3 inhibitors (NaCl, K2 TeO3 , and phenethyl alcohol), and 2 accelerators (sodium pyruvate and glycine). After 4 h of selective cultivation, bacterial detection was accomplished using computer vision. The total analysis time was 5 h. Compared to the Baird-Parker plate count method, which requires 4 to 5 d, this new detection method offers great time savings. Moreover, our novel method had a correlation coefficient of greater than 0.998 when compared with the Baird-Parker plate count method. The detection range for S. aureus was 10 to 10(7) CFU/mL. Our new, rapid detection method for microorganisms in foods has great potential for routine food safety control and microbiological detection applications. © 2014 Institute of Food Technologists®

Chromophore-enhanced bacterial photothermolysis

NASA Astrophysics Data System (ADS)

Huckleby, Jana K.; Morton, Rebecca J.; Bartels, Kenneth E.

1999-06-01

The use of chromophore dyes to enhance the bactericidal effect of laser energy was studied as a means to optimize laser treatment for the decontamination of wound. Using an in vitro study, various concentrations of indocyanine green (ICG), carbon black, and fluorescein were mixed with a suspension of bacteria and plated on tryptic soy agar. Plates were exposed to a laser beam of 10-15 watts for times ranging from 0 to 180 seconds, incubated overnight, and colony counts were performed. Bacteria not mixed with chromophore were used as controls. Six bacterial strains encompassing a range of bacterial types were used: Staphylococcus aureau, Streptococcus pyogenes, Escherichia coli, Pseudomonas aeruginosa, Bacillus cereus spore suspensions, and Clostridium perfringens. Laser treatment alone had no effect on any of the bacteria. Significant killing of gram-positive bacteria, including spores of Bacillus cereus, was observed only with the use of ICG and diode laser energy. No effect was observed using any of the chromophores on the gram-negative bacteria. The results of this study indicate that successful killing of gram-positive bacteria can be achieved using ICG combined with appropriate laser energy and wavelength. Efforts to enhance the susceptibility of gram-negative bacteria to photothermolysis by laser energy were unsuccessful.

Evaluation of different selective media and culturing techniques for the quantification of Campylobacter ssp. from broiler litter.

PubMed

Kiess, A S; Parker, H M; McDaniel, C D

2010-08-01

Poultry is a major reservoir for Campylobacter, the leading cause of foodborne illness in the United States, but how broilers become initially colonized is still under debate. Broiler litter is a potential source, but the best technique for quantifying Campylobacter from litter is still unknown. Therefore, our objectives were to determine if certain media are more selective for quantifying Campylobacter and if enrichment allows for the detection of stressed or viable but nonculturable cells from broiler litter samples. In this trial, 5 media and 2 culturing techniques were used to enumerate Campylobacter from broiler litter. The media used were campy-Line agar (CLA), campy-cefex agar (CCA), modified CCA, Campylobacter agar plates (CAP), and modified charcoal cefoperazone deoxycholate agar. Litter samples were obtained from a commercial broiler house. Each sample was equally divided and diluted 10-fold into peptone, for direct plating, or 4-fold into Campylobacter enrichment broth. Samples diluted in peptone were direct-plated onto each media and incubated under microaerophilic conditions for 48 h at 42 degrees C. Samples diluted in enrichment broth were incubated under the same conditions for 24 h, then further diluted to 10-fold before plating. Plates from enriched samples were incubated for an additional 24 h after plating. After incubation, all plates (direct and enriched) were counted and presumptive positive colonies were confirmed using a Campylobacter latex agglutination kit. Results indicated that there was no difference in the ability of any of the selective media tested to grow Campylobacter. Direct-plated samples had a higher Campylobacter isolation rate compared with enriched samples. The CLA and CAP were able to suppress total bacterial growth better than modified charcoal cefoperazone deoxycholate, modified CCA, and CCA. The CLA and CAP were the only media able to detect total bacterial population shifts over time. In conclusion, it is important before making a final decision on a selective medium to consider the medium's ability to suppress total bacterial growth as well as isolate Campylobacter.

Precise, High-throughput Analysis of Bacterial Growth.

PubMed

Kurokawa, Masaomi; Ying, Bei-Wen

2017-09-19

Bacterial growth is a central concept in the development of modern microbial physiology, as well as in the investigation of cellular dynamics at the systems level. Recent studies have reported correlations between bacterial growth and genome-wide events, such as genome reduction and transcriptome reorganization. Correctly analyzing bacterial growth is crucial for understanding the growth-dependent coordination of gene functions and cellular components. Accordingly, the precise quantitative evaluation of bacterial growth in a high-throughput manner is required. Emerging technological developments offer new experimental tools that allow updates of the methods used for studying bacterial growth. The protocol introduced here employs a microplate reader with a highly optimized experimental procedure for the reproducible and precise evaluation of bacterial growth. This protocol was used to evaluate the growth of several previously described Escherichia coli strains. The main steps of the protocol are as follows: the preparation of a large number of cell stocks in small vials for repeated tests with reproducible results, the use of 96-well plates for high-throughput growth evaluation, and the manual calculation of two major parameters (i.e., maximal growth rate and population density) representing the growth dynamics. In comparison to the traditional colony-forming unit (CFU) assay, which counts the cells that are cultured in glass tubes over time on agar plates, the present method is more efficient and provides more detailed temporal records of growth changes, but has a stricter detection limit at low population densities. In summary, the described method is advantageous for the precise and reproducible high-throughput analysis of bacterial growth, which can be used to draw conceptual conclusions or to make theoretical observations.

Direct quantification of test bacteria in synthetic water-polluted samples by square wave voltammetry and chemometric methods.

PubMed

Carpani, Irene; Conti, Paolo; Lanteri, Silvia; Legnani, Pier Paolo; Leoni, Erica; Tonelli, Domenica

2008-02-28

A home-made microelectrode array, based on reticulated vitreous carbon, was used as working electrode in square wave voltammetry experiments to quantify the bacterial load of Escherichia coli ATCC 13706 and Pseudomonas aeruginosa ATCC 27853, chosen as test microorganisms, in synthetic samples similar to drinking water (phosphate buffer). Raw electrochemical signals were analysed with partial least squares regression coupled to variable selection in order to correlate these values with the bacterial load estimated by aerobic plate counting. The results demonstrated the ability of the method to detect even low loads of microorganisms in synthetic water samples. In particular, the model detects the bacterial load in the range 3-2,020 CFU ml(-1) for E. coli and in the range 76-155,556 CFU ml(-1) for P. aeruginosa.

Evaluation of microbial survival post-incidence on fresh Mozzarella cheese.

PubMed

Ganesan, Balasubramanian; Irish, David A; Brothersen, Carl; McMahon, Donald J

2012-12-01

Commercial fresh Mozzarella cheese is made by direct acidification and is stored dry or in water without salt addition. The cheese has a shelf life of 6 wk, but usually develops an off-flavor and loses textural integrity by 4 wk, potentially due to the lack of salt and high moisture that allow the outgrowth of undesirable bacteria. To understand how microbial incidence affects cheese quality and how incident pathogen-related bacteria are limited by salt level during refrigerated storage, we made fresh Mozzarella cheese with high (2%) and low (0.5%) salt. The high-salt cheese was packaged and stored dry. The low-salt cheese was packaged and stored either dry or in 0.5% salt brine. One portion of cheeses was evaluated for surviving incident microbes by aerobic plate counts, coliform counts, and psychrophilic bacterial counts, of which coliforms and psychrophiles were not detected over 9 wk. Aerobic plate counts remained at 100 to 300 cfu/g up to 2 wk but increased by 1,000- to 10,000-fold between 4 and 6 wk at all salt levels and storage conditions. Other portions of cheeses were inoculated with either Escherichia coli or Enterococcus faecalis, both of which increased by 100-fold over 90 d of storage. Interestingly, E. coli added to the cheese brine first grew in the brine by 100-fold before attaching to the cheese, whereas Ent. faecalis attached to the cheese within 24h and grew only on the cheese. We conclude that incident bacteria, even from similar environments, may attach to cheese curd and survive differently in fresh Mozzarella cheese than in brine. Overall, 2% salt was insufficient to control bacterial growth, and slow-growing, cold- and salt-tolerant bacteria may survive and spoil fresh Mozzarella cheese. Copyright © 2012 American Dairy Science Association. Published by Elsevier Inc. All rights reserved.

Improved methods for the enumeration of heterotrophic bacteria in bottled mineral waters.

PubMed

Ramalho, R; Cunha, J; Teixeira, P; Gibbs, P A

2001-03-01

At this time the European Union regulations require that the heterotrophic plate counts (HPC) of mineral waters be assessed at two recovery temperatures: 22 degrees C for 72 h and 37 degrees C for 24 h. This procedure is time consuming and expensive. Development of new rapid methods for microbiological assessment of the microbial flora in the bottled water is an industry-driven need. The objectives of this work were to develop a method for the HPC that utilises only one recovery temperature and one incubation period and evaluate the use of, the LIVE/DEAD(R) BacLight Bacterial Viability Kit, 5-cyano-2,3-ditotyl tetrazolium chloride (CTC) and impedance methods to enumerate viable bacteria in bottled mineral water. Results showed that incubation at 30 degrees C could be used instead of incubation at 22 degrees C and 37 degrees C. Good correlation exists between counts at 30 degrees C and counts at 22 degrees C (r>0.90) and all the pathogens important in mineral water analyses grow similarly at 30 degrees C and 37 degrees C during 24 h. It was demonstrated that impedance methods might be useful to the mineral water industry as a rapid indicator of microbiological quality of the water. Results obtained with BacLight and CTC were similar to those obtained with plate counts.

Effect of mobile unidirectional air flow unit on microbial contamination of air in standard urologic procedures.

PubMed

Ferretti, Stefania; Pasquarella, Cesira; Fornia, Samanta; Saccani, Elisa; Signorelli, Carlo; Vitali, Pietro; Sansebastiano, Giuliano Ezio

2009-12-01

Infection is one of the most feared complications of surgery. New instrumentation is being developed to reduce deposition of bacteria. We investigated 45 major surgical procedures (21 radical nephrectomies [RN] and 24 radical retropubic prostatectomies [RRP]) in our urology department during 2007. In about one-half of the interventions, an ultraclean air flow mobile (UAF) unit was used. Bacterial sedimentation was evaluated by nitrocellulose membranes placed on the instrument tray and by settle plates positioned at four points in the operating room. In 27 operations, an additional membrane was located near the incision. Bacterial counts on the nitrocellulose membranes during RN were 230 colony-forming units (cfu)/m(2)/h with the UAF unit and 2,254 cfu/m(2)/h without the unit (p = 0.001). During RRP, the values were 288 cfu/m(2)/h and 3,126 cfu/m(2)/h respectively (p = 0.001). The membrane placed near the incision during RN showed a microbial count of 1,235 cfu/m(2)/h with the UAF unit and 5,093 cfu/m(2)/h without the unit (p = 0.002); during RRP, the values were 1,845 cfu/m(2)/h and 3,790 cfu/m(2)/h, respectively (difference not significant). Bacterial contamination detected by settle plates during RN showed a mean value of 2,273 cfu/m(2)/h when the UAF unit was used and 2,054 cfu/m(2)/h without the unit; during RRP, the values were 2,332 cfu/m(2)/h and 2,629 cfu/m(2)/h with and without the UAF unit, respectively (NS). No statistically significant differences were detected in the clinical data registered in patients operated on under standard conditions and while the UAF unit was functioning. The UAF appears able to reduce microbial contamination at the operating table, reaching a bacterial number obtained in ultraclean operating theatres.

Host Immune Response to Bacterial Cyclic Diguanylic Acid (c-di-GMP)

DTIC Science & Technology

2009-07-01

at 4°C. Sera were harvested and kept at 20°C until tested. The institutional ethics committee on animal experimentation of the Faculté des Sciences...On day 5, nonadherent DCs were harvested by gentle pipetting, counted, and plated in fresh medium containing GM-CSF and IL-4 (50 ng/ml each). On day...incubation, cells were pulsed with 1 Ci of [3H]thymidine/well for 18 h and were harvested on filter paper. Proliferative responses were measured as [3H

An evaluation of a partial-walled laminar-flow operating room

PubMed Central

Whyte, W.; Shaw, B. H.; Freeman, M. A. R.

1974-01-01

This paper contains an assessment of the physical performance of a permanently installed down-flow laminar-flow operating room at the London Hospital. This system employs partial walls extending 0·76 m (2·5 ft.) from the ceiling, from which the air is allowed to issue freely downwards at an initial velocity of about 0·4 m./sec. (80 ft./min.). The usefulness of the partial wall, as compared with a free issuing system, was demonstrated and a comparison made with a fully walled system. It was shown that a fully walled system would be more efficient than a partial-walled system as there was a loss in air velocity of about 20-25% with the partial wall due to the nonconstrained flow of air. This loss would be reflected in an increase in airborne bacterial count and would mean that an increase of 20-25% in the air volume would be required to obtain the same conditions as with the full-walled system. Entrainment of contaminated air was demonstrated but it was concluded that this would be of little consequence in the centre of the clean area, i.e. at the wound site. Sterile instruments, etc., however, on the outside of the clean area, would be more liable to airborne contamination. Bacterial and dust airborne counts taken during total hip operations gave a very low average figure (0·3 bacteria/ft.3 or 10·5/m.3) from which we conclude that the system was about 30 times cleaner in terms of airborne bacteria than a well ventilated conventional operating-room. We concluded that although the partial-walled system was slightly less efficacious than a normal full-walled system, the freedom of movement and of communication for the operating team could in some circumstances outweigh this disadvantage. Sound levels were such that normal conversation was possible with little or no awareness of background noise. ImagesFig. 2Fig. 3Plate 2Plate 2Plate 3Plate 3Plate 1 PMID:4529595

Laboratory evaluation of 3M Petrifilms and University of Minnesota Bi-plates as potential on-farm tests for clinical mastitis.

PubMed

McCarron, J L; Keefe, G P; McKenna, S L B; Dohoo, I R; Poole, D E

2009-05-01

The objective was to determine test characteristics and compare 2 potential on-farm culture systems for clinical mastitis, the Minnesota Easy Culture System II Bi-plate and Petrifilm. The tests were evaluated using clinically positive mastitic milk samples (n = 282) to determine their ability to differentiate appropriate treatment groups; all cases that had gram-positive growth were considered treatment candidates (n = 161), whereas cases that grew gram-negative organisms only or yielded no bacterial growth were classified as no treatment (n = 121). For Petrifilm, both undiluted and 1:10 diluted milk samples were used. To create treatment categories, 2 types of Petrifilms were used, Aerobic Count (AC) and Coliform Count (CC). Both Bi-plates and Petrifilms were read after 24 h of incubation. Analysis was conducted at various colony count thresholds for the Petrifilm test system. The combination of Petrifilms that had the highest sensitivity classified a case as gram-negative if there were > or =20 colonies present on the CC. If there were <20 colonies present on the CC and >5 colonies present on the AC, a case would be classified as gram-positive. The Bi-plate had a sensitivity of 97.9% and a specificity of 68.6%. The Petrifilm test system had a sensitivity of 93.8% and a specificity of 70.1%. There was no significant difference in the sensitivities between the tests. All Bi-plates and Petrifilms were read by a laboratory technician and a group of masked readers with limited microbiology training. Kappa values for the masked readers were 0.75 for Bi-plates and 0.84 and 0.86 for AC and CC Petrifilms, respectively. The Bi-plate and Petrifilm were able to successfully categorize clinical cases of mastitis into 2 treatments based on their ability to detect the presence of a gram-positive organism. Neither method had the ability to determine if a sample was contaminated. The results of this study indicate that both tests were able to appropriately categorize cases, which could potentially result in a reduction in the quantity of antibiotics used to treat clinical cases of mastitis.

Bactericidal effects of a high-power, red light-emitting diode on two periodontopathic bacteria in antimicrobial photodynamic therapy in vitro.

PubMed

Umeda, Makoto; Tsuno, Akiko; Okagami, Yoshihide; Tsuchiya, Fumito; Izumi, Yuichi; Ishikawa, Isao

2011-11-01

  Light-emitting diodes have been investigated as new light activators for photodynamic therapy. We investigated the bactericidal effects of high-power, red light-emitting diodes on two periodontopathic bacteria in vitro.   A light-emitting diode (intensity: 1100 mW/cm(2) , peak wavelength: 650 nm) was used to irradiate a bacterial solution for either 10 or 20 s. Bacterial solutions (Porphyromonas gingivalis or Aggregatibacter actinomycetemcomitans) at a concentration of 2.5 × 10(6) c.f.u./mL were mixed with an equal volume of either methylene blue or toluidine blue O (0-20 μg/mL) and added to titer plate wells. The plate wells were irradiated with red light-emitting diode light from a distance of 22 or 40 mm. The contents were diluted, and 50 μL was smeared onto blood agar plates. After 1 week of culturing, bacterial c.f.u. were counted.   The light-emitting diode energy density was estimated to be approximately 4 and 8 J/cm(2) after 10 and 20 s of irradiation, respectively. Red light-emitting diode irradiation for 10 s from a distance of 22 mm, combined with methylene blue at concentrations >10 μg/mL, completely killed Porphyromonas gingivalis and Aggregatibacter actinomycetemcomitans.   High-power, red light-emitting diode irradiation with a low concentration of dye showed effective bactericidal effects against two periodontopathic bacteria. © 2011 Blackwell Publishing Asia Pty Ltd.

Evaluation of the antibacterial activity of a conventional orthodontic composite containing silver/hydroxyapatite nanoparticles.

PubMed

Sodagar, Ahmad; Akhavan, Azam; Hashemi, Ehsan; Arab, Sepideh; Pourhajibagher, Maryam; Sodagar, Kosar; Kharrazifard, Mohammad Javad; Bahador, Abbas

2016-12-01

One of the most important complications of fixed orthodontic treatment is the formation of white spots which are initial carious lesions. Addition of antimicrobial agents into orthodontic adhesives might be a wise solution for prevention of white spot formation. The aim of this study was to evaluate the antibacterial properties of a conventional orthodontic adhesive containing three different concentrations of silver/hydroxyapatite nanoparticles. One hundred and sixty-two Transbond XT composite discs containing 0, 1, 5, and 10 % silver/hydroxyapatite nanoparticles were prepared and sterilized. Antibacterial properties of these composite groups against Streptococcus mutans, Lactobacillus acidophilus, and Streptococcus sanguinis were investigated using three different antimicrobial tests. Disk agar diffusion test was performed to assess the diffusion of antibacterial agent on brain heart infusion agar plate by measuring bacterial growth inhibition zones. Biofilm inhibition test showed the antibacterial capacity of composite discs against resistant bacterial biofilms. Antimicrobial activity of eluted components from composite discs was investigated by comparing the viable counts of bacteria after 3, 15, and 30 days. Composite discs containing 5 and 10 % silver/hydroxyapatite nanoparticles were capable of producing growth inhibition zones for all bacterial types. Results of biofilm inhibition test showed that all of the study groups reduced viable bacterial count in comparison to the control group. Antimicrobial activity of eluted components from composite discs was immensely diverse based on the bacterial type and the concentration of nanoparticles. Transbond XT composite discs containing 5 and 10 % silver/hydroxyapatite nanoparticles produce bacterial growth inhibition zones and show antibacterial properties against biofilms.

Fluorometric determination of the DNA concentration in municipal drinking water.

PubMed Central

McCoy, W F; Olson, B H

1985-01-01

DNA concentrations in municipal drinking water samples were measured by fluorometry, using Hoechst 33258 fluorochrome. The concentration, extraction, and detection methods used were adapted from existing techniques. The method is reproducible, fast, accurate, and simple. The amounts of DNA per cell for five different bacterial isolates obtained from drinking water samples were determined by measuring DNA concentration and total cell concentration (acridine orange epifluorescence direct cell counting) in stationary pure cultures. The relationship between DNA concentration and epifluorescence total direct cell concentration in 11 different drinking water samples was linear and positive; the amounts of DNA per cell in these samples did not differ significantly from the amounts in pure culture isolates. We found significant linear correlations between DNA concentration and colony-forming unit concentration, as well as between epifluorescence direct cell counts and colony-forming unit concentration. DNA concentration measurements of municipal drinking water samples appear to monitor changes in bacteriological quality at least as well as total heterotrophic plate counting and epifluorescence direct cell counting. PMID:3890737

Rapid surface colony counts determination with three new miniaturised techniques.

PubMed

Malik, K A

1977-01-01

Three different miniaturised methods for the rapid surface viable counting are described. The methods were tried in parallel to seven different existing methods (Table 1) for viable counts and were found to be easier, quicker and insome cases more accurate. The techniques require about 10% of the material and time needed for conventional spread-plates method and the results were in no way inferior to that (Table 1 and 2). Mini agar discs were cut aseptically with an especially designed stainless steel agar disc cutter (25 mm internal and 28 mm external diameter, Fig. 1b) or with a test tube of similar diameter. The area of the resulted mini-agar-disc of 25 mm diameter was kept such (about 1/10th of the normal plate) that the ratio of the colony-bearing area to the inoculm remained the same as on big plates in spread-plate-method (Table 2). In normal Petri dishes (about 90 mm diameter) up to seven mini agar discs were possible to cut. Each small agar disc was seperated from the other mini-disc by a distance of at least 6 mm (Fig. 1a). The empty place around the disc was still enlarged during over drying of the plates and during incubation. This created complete isolation from the neighbouring disc. For micro-determination of surface viable counts 10 micronl from each dilution was delivered on a well-dired mini-disc with a piston micropipette. The inoculm was immediately spread on the whole mini-disc with a specially designed flame sterilizable platinum-Mini-spreader (Fig. 2a). No spinning of the plate was needed. Alternatively the dropping pipette and spreader was replaced by a calibrated platinum wire Loop-spreader (Fig. 2b). A loop of 3 mm internal diameter made from a platinum-iridium wire of 0.75 mm thickness proved most useful and carried a drop of 10 micronl. Differences especially in surface tension of various diluting fluids did not influence to drop of this size and no recalibration was needed for water and nutrient broth. The loop was further shaped to Loop-spreader form. From each bacterial suspension 10 micronl were carried and spread on each mini-disc. The method is useful for pathogenic organisms as the loop can readily be flame sterilized. For routine purposes where only approximate numbers of bacteria need to be known a still rapid semiquantitative method was deviced making use of a calibrated stainless steel Stamping-disc (Fig. 2c). A disc of 25mm diameter and 1 mm thickness delivered approximateyl 10 microlitres of supensions and was found to be most useful to stamp seven dilutions on a single plate. In collections and bacteriology laboratories where by conventional methods large number of plates are to be plated and counted the presented techniques could prove most convenient, rapid and economical.

Impact of different-sized laminar air flow versus no laminar air flow on bacterial counts in the operating room during orthopedic surgery.

PubMed

Diab-Elschahawi, Magda; Berger, Jutta; Blacky, Alexander; Kimberger, Oliver; Oguz, Ruken; Kuelpmann, Ruediger; Kramer, Axel; Assadian, Ojan

2011-09-01

This study investigated the influence of the size of unidirectional ceiling distribution systems on counts of viable microorganisms recovered at defined sites in operating room (ORs) and on instrument tables during orthopedic surgery. We compared bacterial sedimentation during 80 orthopedic surgeries. A total of 19 surgeries were performed in ORs with a large (518 cm × 380 cm) unidirectional ceiling distribution (colloquially known as laminar air flow [LAF]) ventilation system, 21 procedures in ORs with a small (380 cm × 120 cm) LAF system, and 40 procedures in ORs with no LAF system. Bacterial sedimentation was evaluated using both settle plates and nitrocellulose membranes. Multivariate linear regression analysis revealed that the colony-forming unit count on nitrocellulose membranes positioned on the instrument table was significantly associated only with the size of the unidirectional LAF distribution system (P < .001), not with the duration of the surgical intervention (P = .753) or with the number of persons present during the surgical intervention (P = .291). Our findings indicate that simply having an LAF ventilation system in place will not provide bacteria-free conditions at the surgical site and on the instrument table. In view of the limited number of procedures studied, our findings require confirmation and further investigations on the ideal, but affordable, size of LAF ventilation systems. Copyright © 2011 Association for Professionals in Infection Control and Epidemiology, Inc. Published by Mosby, Inc. All rights reserved.

Association between bacterial survival and free chlorine concentration during commercial fresh-cut produce wash operation.

PubMed

Luo, Yaguang; Zhou, Bin; Van Haute, Sam; Nou, Xiangwu; Zhang, Boce; Teng, Zi; Turner, Ellen R; Wang, Qin; Millner, Patricia D

2018-04-01

Determining the minimal effective free chlorine (FC) concentration for preventing pathogen survival and cross-contamination during produce washing is critical for developing science- and risk-based food safety practices. The correlation between dynamic FC concentrations and bacterial survival was investigated during commercial washing of chopped Romaine lettuce, shredded Iceberg lettuce, and diced cabbage as pathogen inoculation study during commercial operation is not feasible. Wash water was sampled every 30 min and assayed for organic loading, FC, and total aerobic mesophilic bacteria after chlorine neutralization. Water turbidity, chemical oxygen demand, and total dissolved solids increased significantly over time, with more rapid increases in diced cabbage water. Combined chlorine increased consistently while FC fluctuated in response to rates of chlorine dosing, product loading, and water replenishment. Total bacterial survival showed a strong correlation with real-time FC concentration. Under approximately 10 mg/L, increasing FC significantly reduced the frequency and population of surviving bacteria detected. Increasing FC further resulted in the reduction of the aerobic plate count to below the detection limit (50 CFU/100 mL), except for a few sporadic positive samples with low cell counts. This study confirms that maintaining at least 10 mg/L FC in wash water strongly reduced the likelihood of bacterial survival and thus potential cross contamination of washed produce. Published by Elsevier Ltd.

A Chlorhexidine Solution Reduces Aerobic Organism Growth in Operative Splash Basins in a Randomized Controlled Trial.

PubMed

Lindgren, Kevin E; Pelt, Christopher E; Anderson, Mike B; Peters, Christopher L; Spivak, Emily S; Gililland, Jeremy M

2018-01-01

Despite recommendations against the use of splash basins, due to the potential of bacterial contamination, our observation has been that they continue to be used in operating theaters. In hopes of decontaminating the splash basin, we sought to determine if the addition of chlorhexidine gluconate (CHG) would eliminate aerobic bacterial growth within the splash basin. After Institutional Review Board approval, we began enrollment in a randomized controlled trial comparing 2 splash basin solutions. Splash basins (n = 111) were randomized to either the standard of care (control) solution of sterile water or the experimental solution containing 0.05% CHG. One 20 mL aliquot was taken from the basin at the end of the surgical case and delivered to an independent laboratory. Samples were plated on tryptic soy agar (medium) and incubated at 30°C-35°C to encourage growth. After 48-72 hours, the agar plates were examined for growth and a standard plate count of aerobic cultures was performed. The sterile water group was found to have bacterial growth in 9% of samples compared to no growth in the CHG group (P = .045). The organisms included Micrococcus luteus, Staphylococcus hominis, Gram-variable coccobacilli, and unidentifiable Gram-positive rods. Given the safety and efficacy of a concentration of 0.05% CHG in reducing the bacterial contamination in the operative splash basin, it would seem that if the practice of using a splash basin in the operating theater is to be continued, the addition of an antiseptic solution such as that studied here should be considered. Copyright © 2017 Elsevier Inc. All rights reserved.

Evaluation of antimicrobial activity of selected plant extracts by rapid XTT colorimetry and bacterial enumeration.

PubMed

Al-Bakri, Amal G; Afifi, Fatma U

2007-01-01

The aim of this study was to screen and evaluate the antimicrobial activity of indigenous Jordanian plant extracts, dissolved in dimethylsulfoxide, using the rapid XTT assay and viable count methods. XTT rapid assay was used for the initial screening of antimicrobial activity for the plant extracts. Antimicrobial activity of potentially active plant extracts was further assessed using the "viable plate count" method. Four degrees of antimicrobial activity (high, moderate, weak and inactive) against Bacillus subtilis, Staphylococcus aureus, Escherichia coli and Pseudomonas aeruginosa, respectively, were recorded. The plant extracts of Hypericum triquetrifolium, Ballota undulata, Ruta chalepensis, Ononis natrix, Paronychia argentea and Marrubium vulgare had shown promising antimicrobial activity. This study showed that while both XTT and viable count methods are comparable when estimating the overall antimicrobial activity of experimental substances, there is no strong linear correlation between the two methods.

Microbiological baseline study of poultry slaughtered in provincially inspected abattoirs in Alberta, Canada

PubMed Central

Bohaychuk, Valerie M.; Checkley, Sylvia L.; Gensler, Gary E.; Barrios, Pablo Romero

2009-01-01

Studies to determine baseline levels of microbial contaminants and foodborne bacterial pathogens are needed to evaluate the effectiveness of Hazard Analysis Critical Control Point (HACCP) programs, Good Manufacturing/Production Practices, and various interventions. In 2004 and 2005 poultry carcass rinses from provincially inspected abattoirs in Alberta, Canada, were tested to determine the levels of aerobic plate count bacteria, coliform bacteria, and generic Escherichia coli, the prevalence and levels of Campylobacter spp., and the prevalence of Salmonella spp. and Shiga toxin-producing E. coli (STEC). Samples were collected from 3 high volume and 62 low volume abbatoirs. All samples (1296) were positive for aerobic plate count bacteria, with 98.8% of samples having counts of 100 000 or less colony forming units (CFU)/cm2. Coliform bacteria were isolated from 99.7% of the 1296 carcasses and were recovered at levels of ≤ 1000 CFU/cm2 for 98.3% of the samples. Generic E. coli were recovered from 99.1% of the 1296 carcasses at levels of ≤ 1000 CFU/cm2 for 98.6% of the samples. Seventy five percent of 1234 samples that were tested for Campylobacter were positive; 37.5% of 1295 samples that were tested for Salmonella were positive; and only 2 of 1296 samples tested for STEC were positive (0.15%). PMID:19412397

Phage-based biomolecular filter for the capture of bacterial pathogens in liquid streams

NASA Astrophysics Data System (ADS)

Du, Songtao; Chen, I.-Hsuan; Horikawa, Shin; Lu, Xu; Liu, Yuzhe; Wikle, Howard C.; Suh, Sang Jin; Chin, Bryan A.

2017-05-01

This paper investigates a phage-based biomolecular filter that enables the evaluation of large volumes of liquids for the presence of small quantities of bacterial pathogens. The filter is a planar arrangement of phage-coated, strip-shaped magnetoelastic (ME) biosensors (4 mm × 0.8 mm × 0.03 mm), magnetically coupled to a filter frame structure, through which a liquid of interest flows. This "phage filter" is designed to capture specific bacterial pathogens and allow non-specific debris to pass, eliminating the common clogging issue in conventional bead filters. ANSYS Maxwell was used to simulate the magnetic field pattern required to hold ME biosensors densely and to optimize the frame design. Based on the simulation results, a phage filter structure was constructed, and a proof-in-concept experiment was conducted where a Salmonella solution of known concentration were passed through the filter, and the number of captured Salmonella was quantified by plate counting.

Bacterial toxicity/compatibility of platinum nanospheres, nanocuboids and nanoflowers

PubMed Central

Gopal, Judy; Hasan, Nazim; Manikandan, M.; Wu, Hui-Fen

2013-01-01

For the first time, we have investigated the bacterial toxicity or compatibility properties of Pt nanoparticles (NPs) with different sizes (P1, P2, P3, P4 and P5). The bacterio-toxic or compatible properties of these five different sized Pt NPs with the clinical pathogen, Pseudomonas aeruginosa were explored by many analytical methods such as the conventional plate count method, matrix assisted laser desorption/ionization mass spectrometry (MALDI-MS), fluorescence microscopy and fluorescence sensoring techniques. The results revealed that the 1–3 nm sized (P1 and P2) Pt NPs showed bacterio-toxic properties while the 4–21 nm (P3, P4 and P5) Pt NPs exhibited bacterio-compatible properties. This is the first study which reports the bacterial toxicity of Pt NPs. The information released from this study is significantly important to future clinical, medical, biological and biomedical applications of Pt NPs. PMID:23405274

Identification of novel eubacteria from spent mushroom compost (SMC) waste by DNA sequence typing: ecological considerations of disposal on agricultural land.

PubMed

Watabe, M; Rao, J R; Xu, J; Millar, B C; Ward, R F; Moore, J E

2004-01-01

A small study was undertaken to examine the microbiological characteristics of spent mushroom compost (SMC), which is the major waste by-product of the mushroom industry and which is regularly disposed off by application to agricultural land. The primary aim of this study was to examine SMC for the presence of faecal bacterial pathogens, including Campylobacter spp., Salmonella spp. and Listeria monocytogenes. Secondly it was desirable to quantify bacterial and fungal populations within SMC, and also qualitatively identify the diversity of bacterial populations within SMC, through employment of rDNA PCR and direct sequencing techniques on the culturable microflora. Conventional microbiological analyses of SMC material (n=30) from six commercial operations in both Northern Ireland and the Republic of Ireland, failed to detect Salmonella spp, Listeria spp. or Campylobacter spp. in any of the SMC material examined. Total aerobic plate counts gave a mean count of log10 7.01 colony forming units (cfu) per gram SMC material (range: log10 6.53-7.52 cfu/g). Fungal counts gave a mean count of log(10) 4.57 cfu per gram SMC material (range: log10 3.93-4.98 cfu/g). From a total of greater than 50 colony picks, a total of 12 bacterial morphotypes were identified and were further examined by employment of partial 16S rRNA gene amplification and sequencing techniques, yielding several genera and species, including Bacillus licheniformis, Bacillus subtilis, Klebsiella/Enterobacter sp. Microbacterium sp. Paenibacillus lentimorbus, Pseudomonas mevalonii, Sphingobacterium multivorum and Stenotrophomonas sp. This is the first preliminary report on the microbial diversity of SMC waste and demonstrates the presence of several species that have not been previously described in SMC, in addition to two potentially novel species within the genera Microbacterium and Stenotrophomonas. It is thereby important to examine the ecological microbe-microbe and plant-microbe interactions that are occurring between the native bacterial soil flora and those added annually (theoretically estimated at approximately 10(18) cells) through the application of SMC. Such studies would be beneficial in helping to ascertain the ecological consequences involved in the disposal of SMC waste on agricultural land.

Antimicrobial and anti-adherence activity of various combinations of coffee-chicory solutions on Streptococcus mutans: An in-vitro study

PubMed Central

Sharma, Rama; Reddy, Vamsi Krishna L; Prashant, GM; Ojha, Vivek; Kumar, Naveen PG

2014-01-01

Context: Several studies have demonstrated the activity of natural plants on the dental biofilm and caries development. But few studies on the antimicrobial activity of coffee-based solutions were found in the literature. Further there was no study available to check the antimicrobial effect of coffee solutions with different percentages of chicory in it. Aims: To evaluate the antimicrobial activity of different combinations of coffee-chicory solutions and their anti-adherence effect on Streptococcus mutans to glass surface. Materials and Methods: Test solutions were prepared. For antimicrobial activity testing, tubes containing test solution and culture medium were inoculated with a suspension of S. mutans followed by plating on Brain Heart Infusion (BHI) agar. S. mutans adherence to glass in presence of the different test solutions was also tested. The number of adhered bacteria (CFU/mL) was determined by plating method. Statistical Analysis: Statistical significance was measured using one way ANOVA followed by Tukey's post hoc test. P value < 0.05 was considered statistically significant. Results: Pure chicory had shown significantly less bacterial count compared to all other groups. Groups IV and V had shown significant reduction in bacterial counts over the period of 4 hrs. Regarding anti-adherence effect, group I-IV had shown significantly less adherence of bacteria to glass surface. Conclusions: Chicory exerted antibacterial effect against S. mutans while coffee reduced significantly the adherence of S. mutans to the glass surface. PMID:25328299

In vivo evaluation of the effect of essential oil-containing oral strips on salivary bacteria using the checkerboard method.

PubMed

da Silva, Carina Maciel; Colombo, Andréa Vieira; do Souto, Renata Martins; Colombo, Ana Paula

2005-01-01

The aim of this study was to evaluate the antimicrobial effect of essential oil-containing oral strips on different species of the oral microbiota. Saliva samples were collected from 20 subjects with good oral health, diluted and plated onto blood agar medium. The subjects were asked to place the strip (Listerine PocketPaks) on the tongue allowing it to dissolve. After 30 minutes, new saliva samples were collected again and the plates with the samples were incubated under anaerobic conditions at 37 degrees C for seven days. Colony counts (CFU/mL) were determined for each sample. The colonies on the plates were washed with 1 mL of TE buffer, and the bacterial suspensions were processed for the identification of 24 species by DNA probes and the Checkerboard DNA-DNA hybridization method. Differences in total counts, prevalence, and levels of the species evaluated before and after placement of the strips were determined by Wilcoxon sign rank and Chi-square tests. A modest increase in the total bacterial number in saliva from 1.4 x 10(8) to 1.7 x 10(8) bacterial cells was observed 30 minutes after the strip placement, although this change was not significant (p = 0.632). Most of the species reduced in frequency and/or levels, including the pathogens A. actinomycetemcomitans, C. rectus, E. corrodens, Fusobacterium spp., P. intermedia, and S. noxia, as well as the beneficial species A. meyeri, A. georgia, A. gerencseriae, A. odontolyticus, and P. acnes after strip placement. In contrast, A. viscosus, P. melaninogenica, P. gingivalis, P. micros, Streptococcus spp., T. forsythensis, and V. parvula presented an increase in prevalence and/or levels. These changes were not statistically significant after adjusting for multiple comparisons (p > 0.0022). The use of the essential oil-containing oral strips resulted in a short-term small increase in the total number of salivary microorganisms. In addition, a not significant decrease of certain periodontopathogens, and an increase in species compatible with oral health were observed.

Tolerance of Chemoorganotrophic Bioleaching Microorganisms to Heavy Metal and Alkaline Stresses

PubMed Central

Monballiu, Annick; Cardon, Nele; Tri Nguyen, Minh; Cornelly, Christel; Meesschaert, Boudewijn; Chiang, Yi Wai

2015-01-01

The bioleaching potential of the bacterium Bacillus mucilaginosus and the fungus Aspergillus niger towards industrial residues was investigated by assessing their response towards various heavy metals (including arsenic, cadmium, cobalt, chromium, nickel, lead, and zinc) and elevated pH. The plate diffusion method was performed for each metal to determine the toxicity effect. Liquid batch cultures were set up for more quantitative evaluation as well as for studying the influence of basicity. Growth curves were prepared using bacterial/fungal growth counting techniques such as plate counting, optical density measurement, and dry biomass determination. Cadmium, nickel, and arsenite had a negative influence on the growth of B. mucilaginosus, whereas A. niger was sensitive to cadmium and arsenate. However, it was shown that growth recovered when microorganisms cultured in the presence of these metals were inoculated onto metal-free medium. Based on the findings of the bacteriostatic/fungistatic effect of the metals and the adaptability of the microorganisms to fairly elevated pH values, it is concluded that both strains have potential applicability for further research concerning bioleaching of alkaline waste materials. PMID:26236176

Nondestructive detection of total viable count changes of chilled pork in high oxygen storage condition based on hyperspectral technology

NASA Astrophysics Data System (ADS)

Zheng, Xiaochun; Peng, Yankun; Li, Yongyu; Chao, Kuanglin; Qin, Jianwei

2017-05-01

The plate count method is commonly used to detect the total viable count (TVC) of bacteria in pork, which is timeconsuming and destructive. It has also been used to study the changes of the TVC in pork under different storage conditions. In recent years, many scholars have explored the non-destructive methods on detecting TVC by using visible near infrared (VIS/NIR) technology and hyperspectral technology. The TVC in chilled pork was monitored under high oxygen condition in this study by using hyperspectral technology in order to evaluate the changes of total bacterial count during storage, and then evaluate advantages and disadvantages of the storage condition. The VIS/NIR hyperspectral images of samples stored in high oxygen condition was acquired by a hyperspectral system in range of 400 1100nm. The actual reference value of total bacteria was measured by standard plate count method, and the results were obtained in 48 hours. The reflection spectra of the samples are extracted and used for the establishment of prediction model for TVC. The spectral preprocessing methods of standard normal variate transformation (SNV), multiple scatter correction (MSC) and derivation was conducted to the original reflectance spectra of samples. Partial least squares regression (PLSR) of TVC was performed and optimized to be the prediction model. The results show that the near infrared hyperspectral technology based on 400-1100nm combined with PLSR model can describe the growth pattern of the total bacteria count of the chilled pork under the condition of high oxygen very vividly and rapidly. The results obtained in this study demonstrate that the nondestructive method of TVC based on NIR hyperspectral has great potential in monitoring of edible safety in processing and storage of meat.

Key Issues Concerning Biolog Use for Aerobic and Anaerobic Freshwater Bacterial Community-Level Physiological Profiling

NASA Astrophysics Data System (ADS)

Christian, Bradley W.; Lind, Owen T.

2006-06-01

Bacterial heterotrophy in aquatic ecosystems is important in the overall carbon cycle. Biolog MicroPlates provide information into the metabolic potential of bacteria involved in carbon cycling. Specifically, Biolog EcoPlatesTM were developed with ecologically relevant carbon substrates to allow investigators to measure carbon substrate utilization patterns and develop community-level physiological profiles from natural bacterial assemblages. However, understanding of the functionality of these plates in freshwater research is limited. We explored several issues of EcoPlate use for freshwater bacterial assemblages including inoculum density, incubation temperature, non-bacterial color development, and substrate selectivity. Each of these has various effects on plate interpretation. We offer suggestions and techniques to resolve these interpretation issues. Lastly we propose a technique to allow EcoPlate use in anaerobic freshwater bacterial studies.

A meta-barcoding approach to assess and compare the storage temperature-dependent bacterial diversity of gilt-head sea bream (Sparus aurata) originating from fish farms from two geographically distinct areas of Greece.

PubMed

Parlapani, Foteini F; Michailidou, Sofia; Pasentsis, Konstantinos; Argiriou, Anagnostis; Krey, Grigorios; Boziaris, Ioannis S

2018-08-02

Bacterial diversity of whole gilt-head sea bream (Sparus aurata L. 1758) originating from Ionian and Aegean Sea aquaculture farms and stored at 0 (ice), 4 and 8 °C was determined by 16S rRNA gene amplicon sequencing method using the Illumina's MiSeq platform. The composition of Aerobic Plate Counts (APC) was also monitored by 16S rRNA gene sequencing. The rejection time point of sea bream from either area, as determined by sensory evaluation, was about 14, 6 and 3 days at 0, 4 and 8 °C, respectively. APC was approximately 4.5 log cfu/g at day 0 and ranged from 7.5 to 8.5 log cfu/g at sensory rejection. Culture-depended analysis showed that Pseudomonas and Shewanella were the most abundant microorganisms grown on plates for both seas. Moreover, culture-independent analysis of DNA extracted directly from fish flesh showed that sea bream originating from different geographical areas exhibited different bacterial diversity. Pseudomonas and Psychrobacter were the dominant microorganisms of chill-stored fish from Ionian (apart from 8 °C, where Carnobacterium dominated) and Aegean Sea, respectively. In addition, small changes of storage temperature greatly affected bacterial microbiota of stored fish. Various bacterial species, not detected by conventional microbiological methods, were also revealed through 16S amplicon sequencing. In conclusion, the use of NGS approach is a promising methodology for assessing bacterial diversity of sea bream originating from different geographical areas and stored at various temperatures. Copyright © 2018 Elsevier B.V. All rights reserved.

Automated agar plate streaker: a linear plater on Society for Biomolecular Sciences standard plates.

PubMed

King, Gregory W; Kath, Gary S; Siciliano, Sal; Simpson, Neal; Masurekar, Prakash; Sigmund, Jan; Polishook, Jon; Skwish, Stephen; Bills, Gerald; Genilloud, Olga; Peláez, Fernando; Martín, Jesus; Dufresne, Claude

2006-09-01

Several protocols for bacterial isolation and techniques for aerobic plate counting rely on the use of a spiral plater to deposit concentration gradients of microbial suspensions onto a circular agar plate to isolate colony growth. The advantage of applying a gradient of concentrations across the agar surface is that the original microbiological sample can be applied at a single concentration rather than as multiple serial dilutions. The spiral plater gradually dilutes the sample across a compact area and therefore saves time preparing dilutions and multiple agar plates. Commercial spiral platers are not automated and require manual sample loading. Dispensing of the sample volume and rate of gradients are often very limited in range. Furthermore, the spiral sample application cannot be used with rectangular microplates. Another limitation of commercial spiral platers is that they are useful only for dilute, filtered suspensions and cannot plate suspensions of coarse organic particles therefore precluding the use of many kinds of microorganism-containing substrata. An automated agar plate spreader capable of processing 99 rectangular microplates in unattended mode is described. This novel instrument is capable of dispensing discrete volumes of sample in a linear pattern. It can be programmed to dispense a sample suspense at a uniform application rate or across a decreasing concentration gradient.

HETEROTROPHIC PLATE COUNT BACTERIA - WHAT IS THEIR SIGNIFICANCE IN DRINKING WATER?

EPA Science Inventory

The possible health significance of heterotrophic plate count (HPC) bacteria, also know in earlier terminology as standard plate count (SPC) bacteria, in drinking water has been debated for decades. While the literature documents the universal occurrence of HPC bacteria in soil, ...

A Comparative Evaluation of the Efficacy of Different Caries Excavation Techniques in reducing the Cariogenic Flora: An in vivo Study.

PubMed

Hassan, Afrah Fatima; Yadav, Gunjan; Tripathi, Abhay Mani; Mehrotra, Mridul; Saha, Sonali; Garg, Nishita

2016-01-01

Caries excavation is a noninvasive technique of caries removal with maximum preservation of healthy tooth structure. To compare the efficacy of three different caries excavation techniques in reducing the count of cariogenic flora. Sixty healthy primary molars were selected from 26 healthy children with occlusal carious lesions without pulpal involvement and divided into three groups in which caries excavation was done with the help of (1) carbide bur; (2) polymer bur using slow-speed handpiece; and (3) ultrasonic tip with ultrasonic machine. Samples were collected before and after caries excavation for microbiological analysis with the help of sterile sharp spoon excavator. Samples were inoculated on blood agar plate and incubated at 37°C for 48 hours. After bacterial cultivation, the bacterial count of Streptococcus mutans was obtained. All statistical analysis was performed using SPSS 13 statistical software version. Kruskal-Wallis analysis of variance, Wilcoxon matched pairs test, and Z test were performed to reveal the statistical significance. The decrease in bacterial count of S. mutans before and after caries excavation was significant (p < 0.001) in all the three groups. Carbide bur showed most efficient reduction in cariogenic flora, while ultrasonic tip showed almost comparable results, while polymer bur showed least reduction in cariogenic flora after caries excavation. Hassan AF, Yadav G, Tripathi AM, Mehrotra M, Saha S, Garg N. A Comparative Evaluation of the Efficacy of Different Caries Excavation Techniques in reducing the Cariogenic Flora: An in vivo Study. Int J Clin Pediatr Dent 2016;9(3):214-217.

Sensitive detection of Escherichia coli O157:H7 in food and water by immunomagnetic separation and solid-phase laser cytometry

NASA Technical Reports Server (NTRS)

Pyle, B. H.; Broadaway, S. C.; McFeters, G. A.

1999-01-01

Rapid, direct methods are needed to assess active bacterial populations in water and foods. Our objective was to determine the efficiency of bacterial detection by immunomagnetic separation (IMS) and the compatibility of IMS with cyanoditolyl tetrazolium chloride (CTC) incubation to determine respiratory activity, using the pathogen Escherichia coli O157:H7. Counterstaining with a specific fluorescein-conjugated anti-O157 antibody (FAb) following CTC incubation was used to allow confirmation and visualization of bacteria by epifluorescence microscopy. Broth-grown E. coli O157:H7 was used to inoculate fresh ground beef (<17% fat), sterile 0.1% peptone, or water. Inoculated meat was diluted and homogenized in a stomacher and then incubated with paramagnetic beads coated with anti-O157 specific antibody. After IMS, cells with magnetic beads attached were stained with CTC and then an anti-O157 antibody-fluorescein isothiocyanate conjugate and filtered for microscopic enumeration or solid-phase laser cytometry. Enumeration by laser scanning permitted detection of ca. 10 CFU/g of ground beef or <10 CFU/ml of liquid sample. With inoculated meat, the regression results for log-transformed respiring FAb-positive counts of cells recovered on beads versus sorbitol-negative plate counts in the inoculum were as follows: intercept = 1.06, slope = 0.89, and r2 = 0. 95 (n = 13). The corresponding results for inoculated peptone were as follows: intercept = 0.67, slope = 0.88, and r2 = 0.98 (n = 24). Recovery of target bacteria on beads by the IMS-CTC-FAb method, compared with recovery by sorbitol MacConkey agar plating, yielded greater numbers (beef, 6.0 times; peptone, 3.0 times; water, 2.4 times). Thus, within 5 to 7 h, the IMS-CTC-FAb method detected greater numbers of E. coli O157 cells than were detected by plating. The results show that the IMS-CTC-FAb technique with enumeration by either fluorescence microscopy or solid-phase laser scanning cytometry gave results that compared favorably with plating following IMS.

Do bacterial cell numbers follow a theoretical Poisson distribution? Comparison of experimentally obtained numbers of single cells with random number generation via computer simulation.

PubMed

Koyama, Kento; Hokunan, Hidekazu; Hasegawa, Mayumi; Kawamura, Shuso; Koseki, Shigenobu

2016-12-01

We investigated a bacterial sample preparation procedure for single-cell studies. In the present study, we examined whether single bacterial cells obtained via 10-fold dilution followed a theoretical Poisson distribution. Four serotypes of Salmonella enterica, three serotypes of enterohaemorrhagic Escherichia coli and one serotype of Listeria monocytogenes were used as sample bacteria. An inoculum of each serotype was prepared via a 10-fold dilution series to obtain bacterial cell counts with mean values of one or two. To determine whether the experimentally obtained bacterial cell counts follow a theoretical Poisson distribution, a likelihood ratio test between the experimentally obtained cell counts and Poisson distribution which parameter estimated by maximum likelihood estimation (MLE) was conducted. The bacterial cell counts of each serotype sufficiently followed a Poisson distribution. Furthermore, to examine the validity of the parameters of Poisson distribution from experimentally obtained bacterial cell counts, we compared these with the parameters of a Poisson distribution that were estimated using random number generation via computer simulation. The Poisson distribution parameters experimentally obtained from bacterial cell counts were within the range of the parameters estimated using a computer simulation. These results demonstrate that the bacterial cell counts of each serotype obtained via 10-fold dilution followed a Poisson distribution. The fact that the frequency of bacterial cell counts follows a Poisson distribution at low number would be applied to some single-cell studies with a few bacterial cells. In particular, the procedure presented in this study enables us to develop an inactivation model at the single-cell level that can estimate the variability of survival bacterial numbers during the bacterial death process. Copyright © 2016 Elsevier Ltd. All rights reserved.

Effect of the implementation of HACCP on the microbiological quality of meals at a university restaurant.

PubMed

Cenci-Goga, B T; Ortenzi, R; Bartocci, E; Codega de Oliveira, A; Clementi, F; Vizzani, A

2005-01-01

A study was conducted to evaluate the microbiological quality, including total mesophilic counts and markers of bacteriological hygiene, as indicator of food safety of three categories of the most consumed meals in a university restaurant, before and after implementation of the HACCP system and personnel training. Cold gastronomy products, cooked warm-served products, and cooked cold-served products were tested for bacterial contamination. Throughout the experiment, 894 samples were examined for total counts of aerobic bacteria, counts of indicator organisms (coliform organisms and Escherichia coli) and pathogens (Staphylococcus aureus, Bacillus cereus, Salmonella spp., and Listeria monocytogenes). Implementation of the HACCP system, together with training in personnel hygiene, good manufacturing practices, and cleaning and sanitation procedures, resulted in lower aerobic plate counts and a lower incidence of S. aureus, coliform organisms, E. coli, and B. cereus, whereas Salmonella spp. and L. monocytogenes were not found in all samples studied. The microbial results of this study demonstrate that personnel training together with HACCP application contributed to improve the food safety of meals served in the restaurant studied.

Phage-mediated counting by the naked eye of miRNA molecules at attomolar concentrations in a Petri dish.

PubMed

Zhou, Xin; Cao, Peng; Zhu, Ye; Lu, Wuguang; Gu, Ning; Mao, Chuanbin

2015-10-01

The ability to count biomolecules such as cancer-biomarker miRNAs with the naked eye is seemingly impossible in molecular diagnostics. Here, we show an ultrasensitive naked-eye-counting strategy for quantifying miRNAs by employing T7 phage-a bacteria-specific virus nanoparticle-as a surrogate. The phage is genetically engineered to become fluorescent and capable of binding a miRNA-capturing gold nanoparticle (GNP) in a one-to-one manner. Target miRNAs crosslink the resultant phage-GNP couple and miRNA-capturing magnetic microparticles, forming a sandwich complex containing equimolar phage and miRNA. The phage is then released from the complex and developed into one macroscopic fluorescent plaque in a Petri dish by plating it in a host bacterial medium. Counting the plaques by the naked eye enables the quantification of miRNAs with detection limits of ∼3 and ∼5 aM for single-target and two-target miRNAs, respectively. This approach offers ultrasensitive and convenient quantification of disease biomarkers by the naked eye.

Phage-mediated counting by the naked eye of miRNA molecules at attomolar concentrations in a Petri dish

NASA Astrophysics Data System (ADS)

Zhou, Xin; Cao, Peng; Zhu, Ye; Lu, Wuguang; Gu, Ning; Mao, Chuanbin

2015-10-01

The ability to count biomolecules such as cancer-biomarker miRNAs with the naked eye is seemingly impossible in molecular diagnostics. Here, we show an ultrasensitive naked-eye-counting strategy for quantifying miRNAs by employing T7 phage--a bacteria-specific virus nanoparticle--as a surrogate. The phage is genetically engineered to become fluorescent and capable of binding a miRNA-capturing gold nanoparticle (GNP) in a one-to-one manner. Target miRNAs crosslink the resultant phage-GNP couple and miRNA-capturing magnetic microparticles, forming a sandwich complex containing equimolar phage and miRNA. The phage is then released from the complex and developed into one macroscopic fluorescent plaque in a Petri dish by plating it in a host bacterial medium. Counting the plaques by the naked eye enables the quantification of miRNAs with detection limits of ~3 and ~5 aM for single-target and two-target miRNAs, respectively. This approach offers ultrasensitive and convenient quantification of disease biomarkers by the naked eye.

Determination of viability of Aeromonas hydrophila in increasing concentrations of sodium chloride at different temperatures by flow cytometry and plate count technique.

PubMed

Pianetti, Anna; Manti, Anita; Boi, Paola; Citterio, Barbara; Sabatini, Luigia; Papa, Stefano; Rocchi, Marco Bruno Luigi; Bruscolini, Francesca

2008-10-31

Aeromonads in waters and foods can represent a risk to human health. Factors such as sodium chloride concentration and temperature can affect growth and viability of several food and water-borne pathogens. The behaviour of an Aeromonas hydrophila strain in the presence of 1.7%, 3.4% and 6% NaCl concentrations at 24 degrees C and 4 degrees C was studied over a 188 day period. Viability and membrane potential were assessed by flow cytometry; growth was evaluated by plate count technique. Flow cytometry evidenced that A. hydrophila retained viability over the period although varying according to temperature and salt concentrations. Colony Forming Units were generally lower in number than viable cells especially in the presence of 6% NaCl, indicating the occurrence of stressed cells which maintain metabolic activity yet are not able to grow on agar plates. In conclusion, A. hydrophila showed a long-term halotolerance even at elevated (6%) NaCl concentrations and a lesser sensitivity to salt at low temperature; therefore, low temperature and salt, which are two important factors limiting bacterial growth, do not assure safety in the case of high initial contamination. Finally, cytometry appears a valid tool for the rapid detection of the viability of pathogenic bacteria in food and environmental matrices to control and prevent health risks.

[A comparison between 2 different methods for calculating the percentage of anaerobic microorganisms in the subgingival microbial flora].

PubMed

Petti, S; Renzini, G

1994-03-01

The percentage of anaerobic micro-organisms in the subgingival microflora represents a simple microbiological index which not only refers to the state but also the risks of periodontal health. The present study aimed to compare two different methods of calculating this index. The study was performed in 45 subjects with moderate gingivitis provoked by the previous application of dental fixtures anchored to both arches. A sample of subgingival microflora was collected from each patient at the level of the vestibular gingival sulcus of the first upper right molar. This was then vortexed, diluted and inoculated in three series of plates. It was chosen to use Walker's culture medium. The total bacterial count was evaluated by incubating the first series of plates in anaerobiosis; the anaerobic bacterial was calculated by subtracting from the total the of facultative aerobic-anaerobic micro-organisms, which in turn was obtained using two methods: the first (method AE) consisted of incubating another series of plates in aerobiosis; the second (method M) involved incubating the last series of plates in anaerobiosis, and adding metronidazole to the culture medium in a solution of 2.5 mg/l. The plates were then kept at 37 degrees C for seven days. The mean percentage of anaerobic microorganisms, given by the percentage ratio between anaerobic and total, relating to the 45 cases studied, was as follows: using method AE: 57.8 +/- 26.3%, and using method M: 40.2 +/- 27.2%. Both figures come close to that proposed and calculated using a much more sophisticated method by Slots, namely 41.5 +/- 19.2% in the event of gingivitis.(ABSTRACT TRUNCATED AT 250 WORDS)

Role of mono- and oligosaccharides from FOS as stabilizing agents during freeze-drying and storage of Lactobacillus delbrueckii subsp. bulgaricus.

PubMed

Romano, Nelson; Schebor, Carolina; Mobili, Pablo; Gómez-Zavaglia, Andrea

2016-12-01

The aim of this work was to assess the role of mono- and oligosaccharides present in fructo-oligosaccharides (FOS) mixtures as protective agents during freeze-drying and storage of Lactobacillus delbrueckii subsp. bulgaricus CIDCA 333. Different FOS mixtures were enzymatically obtained from sucrose and further purified by removing the monosaccharides produced as secondary products. Their glass transition temperatures (T g ) were determined at 11, 22 and 33% relative humidity (RH). Bacterial cultures were freeze-dried in the presence of 20% w/v solutions of the studied FOS. Their protective effect during freeze-drying was assessed by bacterial plate counting, and by determining the lag time from growth kinetics and the uptake of propidium iodide (PI). Plate counting during bacterial storage at 4°C, and 11, 22 and 33% RH for 80days completed this rational analysis of the protective effect of FOS. Purification of FOS led to an increase of T g in all the conditions assayed. Microorganisms freeze-dried in the presence of non-purified FOS were those with the shortest lag times. Bacteria freeze-dried with pure or commercial FOS (92% of total FOS) showed larger lag times (8.9-12.6h). The cultivability of microorganisms freeze-dried with non-purified FOS and with sucrose was not significantly different from that of bacteria before freeze-drying (8.74±0.14logCFU/mL). Pure or commercial FOS were less efficient in protecting bacteria during freeze-drying. All the protectants prevented membrane damage. The cultivability of bacteria freeze-dried with FOS decayed <1logarithmicunit after 80days of storage at 11% RH. When storing at 22 and 33% RH, pure and commercial FOS were those that best protected bacteria, and FOS containing monosaccharides were less efficient. The effect of FOS on bacterial protection is the result of a balance between monosaccharides, sucrose and larger FOS in the mixtures: the smallest sugars are more efficient in protecting lipid membranes, and the larger ones favor the formation of vitreous states. Copyright © 2016 Elsevier Ltd. All rights reserved.

Veggie ISS Validation Test Results and Produce Consumption

NASA Technical Reports Server (NTRS)

Massa, Gioia; Hummerick, Mary; Spencer, LaShelle; Smith, Trent

2015-01-01

The Veggie vegetable production system flew to the International Space Station (ISS) in the spring of 2014. The first set of plants, Outredgeous red romaine lettuce, was grown, harvested, frozen, and returned to Earth in October. Ground control and flight plant tissue was sub-sectioned for microbial analysis, anthocyanin antioxidant phenolic analysis, and elemental analysis. Microbial analysis was also performed on samples swabbed on orbit from plants, Veggie bellows, and plant pillow surfaces, on water samples, and on samples of roots, media, and wick material from two returned plant pillows. Microbial levels of plants were comparable to ground controls, with some differences in community composition. The range in aerobic bacterial plate counts between individual plants was much greater in the ground controls than in flight plants. No pathogens were found. Anthocyanin concentrations were the same between ground and flight plants, while antioxidant and phenolic levels were slightly higher in flight plants. Elements varied, but key target elements for astronaut nutrition were similar between ground and flight plants. Aerobic plate counts of the flight plant pillow components were significantly higher than ground controls. Surface swab samples showed low microbial counts, with most below detection limits. Flight plant microbial levels were less than bacterial guidelines set for non-thermostabalized food and near or below those for fungi. These guidelines are not for fresh produce but are the closest approximate standards. Forward work includes the development of standards for space-grown produce. A produce consumption strategy for Veggie on ISS includes pre-flight assessments of all crops to down select candidates, wiping flight-grown plants with sanitizing food wipes, and regular Veggie hardware cleaning and microbial monitoring. Produce then could be consumed by astronauts, however some plant material would be reserved and returned for analysis. Implementation of this plan is a step toward developing pick-and-eat food production to supplement the packaged diet on ISS and for future exploration missions where plants could make up a larger portion of the diet. Supported by NASA Space Biology Program.

Efficacy of on-farm use of ultraviolet light for inactivation of bacteria in milk for calves.

PubMed

Gelsinger, S L; Heinrichs, A J; Jones, C M; Van Saun, R J; Wolfgang, D R; Burns, C M; Lysczek, H R

2014-05-01

Ultraviolet light is being employed for bacterial inactivation in milk for calves; however, limited evidence is available to support the claim that UV light effectively inactivates bacteria found in milk. Thus, the objective of this observational study was to investigate the efficacy of on-farm UV light treatment in reducing bacteria populations in waste milk used for feeding calves. Samples of nonsaleable milk were collected from 9 Pennsylvania herds, twice daily for 15 d, both before and after UV light treatment (n=60 samples per farm), and analyzed for standard plate count, coliforms, noncoliform, gram-negative bacteria, environmental and contagious streptococci, coagulase-negative staphylococci, Streptococcus agalactiae, Staphylococcus aureus count, and total solids percentage, and log reduction and percentage log reduction were calculated. Data were analyzed using the mixed procedure in SAS. In all bacteria types, samples collected after UV treatment contained significantly fewer bacteria compared with samples collected before UV treatment. Weighted least squares means for log reduction (percentage log reduction) were 1.34 (29%), 1.27 (58%), 1.48 (53%), 1.85 (55%), 1.37 (72%), 1.92 (63%), 1.07 (33%), and 1.67 (82%) for standard plate count, coliforms, noncoliform, gram-negative bacteria, environmental and contagious streptococci, Strep. agalactiae, coagulase-negative staphylococci, and Staph. aureus, respectively. A percentage log reduction greater than 50% was achieved in 6 of 8 bacteria types, and 43 and 94% of samples collected after UV treatment met recommended bacterial standards for milk for feeding calves. Based on these results, UV light treatment may be effective for some, but not all bacteria types found in nonsaleable waste milk. Thus, farmers should take into account the bacteria types that may need to be reduced when considering the purchase of a UV-treatment system. Copyright © 2014 American Dairy Science Association. Published by Elsevier Inc. All rights reserved.

[Harmonization of microbiologicaland parasitological indices of epidemic safety of drinking water with the international requirements].

PubMed

Ivanova, L V; Artemova, T Z; Gipp, E K; Zagaĭnova, A V; Maksimkina, T N; Krasniak, A V; Korneĭchuk, S S; Shustova, S S

2013-01-01

For the purpose of harmonization of microbiological and parasitological indices and benchmarks there was performed the comparative analysis of the requirements for the quality of drinking water in respect of the epidemic safety on the basic regulations of Russia, the Directive Council of the European Union EU, WHO, the United States, Canada, Australia, Finland, Sweden, Brazil, France, Japan and China. As a result, there were revealed the priority bacteriological, virological and parasitological parameters: E. coli--indicator of recent fecal contamination, coliforms, heterotrophic bacteria colony count (Heterotrophic plate count), which is in the water legislation of the Russian Federation is characterized as total bacterial count (TBC), being an integral index of the quality of wastewater treatment technologies and hygienic condition of the water supply systems, coliphages as an indicator of viral contamination. In the Guidelines for drinking-water quality control, WHO and a set of countries there is recommended a more wide range of indicators: enterococci, Clostridium perfringens, Pseudomonas aeruginosa, enteroviruses, parasitological indices. With aim of harmonization of the requirements for the quality of drinking water in the Russian Federation with international approaches to the revision of the Sanitary Regulations and Norms (SanPin) 2.1.4.1074 into the project there are introduced priority indicator parameters of bacterial, viral and parasitic contamination of water, evidence-based guidelines.

Water quality problems associated with intermittent water supply.

PubMed

Tokajian, S; Hashwa, F

2003-01-01

A controlled study was conducted in Lebanon over a period of 12 months to determine bacterial regrowth in a small network supplying the Beirut suburb of Naccache that had a population of about 3,000. The residential area, which is fed by gravity, is supplied twice a week with chlorinated water from two artesian wells of a confined aquifer. A significant correlation was detected between the turbidity and the levels of heterotrophic plate count bacteria (HPC) in the samples from the distribution network as well as from the artesian wells. However, a negative significant correlation was found between the temperature and the HPC count in the samples collected from the source. A statistically significant increase in counts, possibly due to regrowth, was repeatedly established between two sampling points lying on a straight distribution line but 1 km apart. Faecal coliforms were detected in the source water but none in the network except during a pipe breakage incident with confirmed Escherichia coli reaching 40 CFU/100 mL. However, coliforms such as Citrobacter freundii, Enterobacter agglomerans, E. cloacae and E. skazakii were repeatedly isolated from the network, mainly due to inadequate chlorination. A second controlled study was conducted to determine the effect of storage on the microbial quality of household storage tanks (500 L), which were of two main types - galvanized cast iron and black polyethylene. The mean bacterial count increased significantly after 7 d storage in both tank types. A significant difference was found in the mean HPC/mL between the winter and the summer. Highest counts were found April-June although the maximum temperature was reported later in the summer. A positive correlation was established between the HPC/mL and pH, temperature and storage time.

Quantitative Urine Culture Method Using a Plastic „Paddle” Containing Dual Media

PubMed Central

Craig, William A.; Kunin, Calvin M.

1972-01-01

A new dip-inoculum method for quantitative urine culture is described which utilizes a dual-chambered plastic „paddle” housing both a general purpose and differential medium. Comparative bacterial counts of 1,000 clinical specimens using the pour plate and this device were identical in 82.9% and within a factor of five in 95.6%. The „paddle” detected all but 19 of 258 specimens (92.6%) with 100,000 or greater colonies per ml. This simple, convenient method should allow more extensive use of quantitative urine culture in the diagnosis and follow-up of patients with urinary tract infections in office practice. It should not be considered as a substitute for the more definitive pour plate method or for standard methods for characterization of bacteriological species when more exact information is required. PMID:4555636

Impact of solar radiation exposure on phyllosphere bacterial community of red-pigmented baby leaf lettuce.

PubMed

Truchado, Pilar; Gil, M Isabel; Reboleiro, Patricia; Rodelas, Belén; Allende, Ana

2017-09-01

Solar radiation has been identified as a stress factor affecting phyllosphere associated bacteria colonization and survival during primary production. In the present study, the impact of different solar radiation doses on the phyllosphere microbiota of red-pigmented baby leaf lettuce cultivated in open field under commercial conditions was evaluated. Four weeks before harvest, the growing field was divided into four plots; each one was consecutively covered with one-week-interval with a light-excluding plastic to reduce the sunlight exposure. Four different solar radiation treatments were generated and cumulative photosynthetically active radiation (PAR) was used to differentiate treatments as follows: 4889 ± 428 μmol/m 2 /s (uncovered), 4265 ± 356 μmol/m 2 /s (covered for 1 week), 3602 ± 225 μmol/m 2 /s (covered for 2 weeks) and 3115 ± 313 μmol/m 2 /s (covered for 3 weeks). The size and composition of the phyllosphere bacterial community were determined by cultivation-depended (plate count) and independent (qPCR) techniques. Exposure to decreased levels of cumulative PAR did not produce significant differences in total bacterial community size, regardless of the chosen quantification techniques. However, total bacteria size quantified by qPCR was around 3.5 orders of magnitude higher than those obtained by plate count. The observed differences between cultivation-depended and independent techniques could be attributed to the presence of non-viable or viable but non-culturable (VBNC) bacteria. The bacterial community structure was analyzed using temperature gradient gel electrophoresis (TGGE), and significant differences were detected when the four solar treatment were compared. A qPCR approach was applied to the quantification of specific bacterial phyla and classes, previously identified in the phyllosphere of plants available literature, confirming that Proteobacteria, Bacteroidetes, Actinobacterias and Firmicutes were the most abundantly represented phyla in lettuce. Treatment comparison revealed higher proportions of Gammaproteobacteria as opposed to the Betaproteobacteria on the lettuce exposed to the lowest cumulative PAR dose (3115 ± 313 μmol/m 2 /s). The obtained results demonstrated that the solar radiation is a relevant environmental factor influencing the relative abundance of specific-groups of phyllosphere-associated bacteria in pigmented baby leaf lettuce. Copyright © 2017 Elsevier Ltd. All rights reserved.

Biogenic amine survey and organoleptic changes in fresh, stored, and temperature-abused bluefish (Pomatomus saltatrix).

PubMed

Gingerich, T M; Lorca, T; Flick, G J; Pierson, M D; McNair, H M

1999-09-01

Changes in histamine, putrescine, and cadaverine concentrations in bluefish filets (Pomatomus saltatrix) stored at 5, 10, and 15 degrees C were determined using high-performance liquid chromatography. An organoleptic assessment was conducted simultaneously with the biogenic amine analyses. The histamine levels found in fresh bluefish obtained from wholesale seafood distributors ranged between <1 ppm and 99 with an average of 39 ppm. Putrescine and cadaverine were not found in fresh bluefish. Fish fillets stored at each of the three temperatures developed histamine. The greatest accumulation of histamine was observed in fish stored at 15 degrees C, which developed histamine levels as high as 2,200 ppm. Putrescine levels increased at each temperature during storage. Cadaverine was present only in uninoculated bluefish stored at 15 degrees C. Histamine achieved higher levels in bluefish pieces inoculated with Morganella morganii, which demonstrates that bluefish support bacterial histamine formation. Histamine levels at each temperature exceeded the 50-ppm advisory level established by the Food and Drug Administration before 100% sensory rejection. Standard plate counts increased during storage of fish at all temperatures, but the correlation between histamine levels and standard plate count was not significant.

Monitoring Microbial Numbers in Food by Density Centrifugation

PubMed Central

Basel, Richard M.; Richter, Edward R.; Banwart, George J.

1983-01-01

Some foods contain low numbers of microbes that may be difficult to enumerate by the plate count method due to small food particles that interfere with the counting of colonies. Ludox colloidal silicon was coated with reducing agents to produce a nontoxic density material. Food homogenates were applied to a layered 10 and 80% mixture of modified Ludox and centrifuged at low speed. The top and bottom of the tube contained the food material, and the Ludox-containing portion was evaluated by conventional pour plate techniques. Plate counts of the Ludox mixture agreed with plate counts of the food homogenate alone. The absence of small food particles from pour plates resulted in a plate that was more easily read than pour plates of the homogenate alone. Modified Ludox was evaluated for its effect on bacteria at 4°C during a 24-h incubation period. No inhibition was observed. This method is applicable to food products, such as doughnuts, spices, tomato products, and meat, in which small food particles often interfere with routine plate counts or low dilution may inhibit colony formation. Inhibitory substances can be removed from spices, resulting in higher counts. Ludox is more economical than similar products, such as Percoll. Modified Ludox is easily rendered nontoxic by the addition of common laboratory reagents. In addition, the mixture is compatible with microbiological media. PMID:6303217

Influence of pre- and post-usage flushing frequencies on bacterial water quality of non-touch water fittings.

PubMed

Suchomel, Miranda; Diab-Elschahawi, Magda; Kundi, Michael; Assadian, Ojan

2013-08-30

Non-touch fittings have been reported to be susceptible for Pseudomonas aeruginosa accumulation. A number of factors may contribute to this, including the frequency of usage, duration of water stagnation, or presence of plastic materials. Programmable non-touch fittings are appearing which allow regular automated post-flushing with cold water to prevent water stagnation. However, the ideal duration of post-flushing is unknown as well as the effect of pre-rinsing with cold water before use. Eight non-touch fittings with brass valve blocks were mounted on a mobile test sink and connected to the same central water pipe source, differing only in presence or absence of water connection pipes, length of connection pipe, frequency of usage, and time intervals for pre- and post-usage water flush. The total bacteria colony-forming unit (cfu) counts were obtained by the spread plate technique. Low frequency of water use in combination with a long stagnating water column resulted in high bacterial cfu counts. Post-usage flushing for 2 seconds did not differ from no flushing. Flushing for 10 seconds with cold water after use or 30 seconds flush before use were both the most effective measures to prevent non-touch fittings from biofilm formation over a period of 20 weeks. Further improvements in water fitting technology could possibly solve the problem of bacterial water contamination in health care settings.

Inactivation of selected bacterial pathogens in dairy cattle manure by mesophilic anaerobic digestion (balloon type digester).

PubMed

Manyi-Loh, Christy E; Mamphweli, Sampson N; Meyer, Edson L; Okoh, Anthony I; Makaka, Golden; Simon, Michael

2014-07-14

Anaerobic digestion of animal manure in biogas digesters has shown promise as a technology in reducing the microbial load to safe and recommended levels. We sought to treat dairy manure obtained from the Fort Hare Dairy Farm by investigating the survival rates of bacterial pathogens, through a total viable plate count method, before, during and after mesophilic anaerobic digestion. Different microbiological media were inoculated with different serial dilutions of manure samples that were withdrawn from the biogas digester at 3, 7 and 14 day intervals to determine the viable cells. Data obtained indicated that the pathogens of public health importance were 90%-99% reduced in the order: Campylobacter sp. (18 days) < Escherichia coli sp. (62 days) < Salmonella sp. (133 days) from a viable count of 10.1 × 103, 3.6 × 105, 7.4 × 103 to concentrations below the detection limit (DL = 102 cfu/g manure), respectively. This disparity in survival rates may be influenced by the inherent characteristics of these bacteria, available nutrients as well as the stages of the anaerobic digestion process. In addition, the highest p-value i.e., 0.957 for E. coli showed the statistical significance of its model and the strongest correlation between its reductions with days of digestion. In conclusion, the results demonstrated that the specific bacterial pathogens in manure can be considerably reduced through anaerobic digestion after 133 days.

The importance of the viable but non-culturable state in human bacterial pathogens

PubMed Central

Li, Laam; Mendis, Nilmini; Trigui, Hana; Oliver, James D.; Faucher, Sebastien P.

2014-01-01

Many bacterial species have been found to exist in a viable but non-culturable (VBNC) state since its discovery in 1982. VBNC cells are characterized by a loss of culturability on routine agar, which impairs their detection by conventional plate count techniques. This leads to an underestimation of total viable cells in environmental or clinical samples, and thus poses a risk to public health. In this review, we present recent findings on the VBNC state of human bacterial pathogens. The characteristics of VBNC cells, including the similarities and differences to viable, culturable cells and dead cells, and different detection methods are discussed. Exposure to various stresses can induce the VBNC state, and VBNC cells may be resuscitated back to culturable cells under suitable stimuli. The conditions that trigger the induction of the VBNC state and resuscitation from it are summarized and the mechanisms underlying these two processes are discussed. Last but not least, the significance of VBNC cells and their potential influence on human health are also reviewed. PMID:24917854

Blue light (470 nm) effectively inhibits bacterial and fungal growth.

PubMed

De Lucca, A J; Carter-Wientjes, C; Williams, K A; Bhatnagar, D

2012-12-01

Blue light (470 nm) LED antimicrobial properties were studied alone against bacteria and with or without the food grade photosensitizer, erythrosine (ERY) against filamentous fungi. Leuconostoc mesenteroides (LM), Bacillus atrophaeus (BA) or Pseudomonas aeruginosa (PA) aliquots were exposed on nutrient agar plates to Array 1 (AR1, 0·2 mW cm(-2)) or Array 2 (AR2, 80 mW cm(-2)), which emitted impure or pure blue light (0-300 J cm(-2)), respectively. Inoculated control (room light only) plates were incubated (48 h) and colonies enumerated. The antifungal properties of blue light combined with ERY (11·4 and 22·8 μmol l(-1)) on Penicillium digitatum (PD) and Fusarium graminearum (FG) conidia were determined. Conidial controls consisted of: no light, room light-treated conidia and ERY plus room light. Light-treated (ERY + blue light) conidial samples were exposed only to AR2 (0-100 J cm(-2)), aliquots spread on potato dextrose agar plates, incubated (48 h, 30°C) and colonies counted. Blue light alone significantly reduced bacterial and FG viability. Combined with ERY, it significantly reduced PD viability. Blue light is lethal to bacteria and filamentous fungi although effectiveness is dependent on light purity, energy levels and microbial genus. Light from two arrays of different blue LEDs significantly reduced bacterial (Leuconostoc mesenteroides, Bacillus atrophaeus and Pseudomonas aeruginosa) viabilities. Significant in vitro viability loss was observed for the filamentous fungi, Penicillium digitatum and Fusarium graminearum when exposed to pure blue light only plus a photosensitizer. F. graminearum viability was significantly reduced by blue light alone. Results suggest that (i) the amount of significant loss in bacterial viability observed for blue light that is pure or with traces of other wavelengths is genus dependent and (ii) depending on fungal genera, pure blue light is fungicidal with or without a photosensitizer. © 2012 The Society for Applied Microbiology.

Detecting swift fox: Smoked-plate scent stations versus spotlighting

Treesearch

Daniel W. Uresk; Kieth E. Severson; Jody Javersak

2003-01-01

We compared two methods of detecting presence of swift fox: smoked-plate scent stations and spotlight counts. Tracks were counted on ten 1-mile (1.6-km) transects with bait/tracking plate stations every 0.1 mile (0.16 km). Vehicle spotlight counts were conducted on the same transects. Methods were compared with Spearman's rank order correlation. Repeated measures...

Fluoridated elastomers: effect on the microbiology of plaque.

PubMed

Benson, Philip E; Douglas, C W Ian; Martin, Michael V

2004-09-01

The objective of this study was to investigate the effect of fluoridated elastomeric ligatures on the microbiology of local dental plaque in vivo. This randomized, prospective, longitudinal, clinical trial had a split-mouth crossover design. The subjects were 30 patients at the beginning of their treatment with fixed orthodontic appliances in the orthodontic departments of the Liverpool and the Sheffield dental hospitals in the United Kingdom. The study consisted of 2 experimental periods of 6 weeks with a washout period between. Fluoridated elastomers were randomly allocated at the first visit to be placed around brackets on tooth numbers 12, 11, 33 or 22, 21, 43. Nonfluoridated elastomers were placed on the contralateral teeth. Standard nonantibacterial fluoridated toothpaste and mouthwash were supplied. After 6 weeks (visit 2), the elastomers were removed, placed in transport media, and plated on agar within 2 hours. Nonfluoridated elastomers were placed on all brackets for 1 visit to allow for a washout period. At visit 3, fluoridated elastomers were placed on the teeth contralateral to those that received them at visit 1. At visit 4, the procedures at visit 2 were repeated. Samples were collected on visits 2 and 4. A logistic regression was performed, with the presence or absence of streptococcal or anaerobic growth as the dependent variable. A mixed-effects analysis of variance was carried out with the percentage of streptococcal or anaerobic bacterial count as the dependent variable. The only significant independent variables were the subject variable (P =<.001) for the percentage of streptococcal and anaerobic bacterial count and the visit variable for the percentage of streptococcal count (P =<.001). The use of fluoridated or nonfluoridated elastomers was not significant for percentage of either streptococcal (P =.288) or anaerobic count (P =.230). Fluoridated elastomers are not effective at reducing local streptococcal or anaerobic bacterial growth after a clinically relevant time in the mouth.

High level bacterial contamination of secondary school students' mobile phones.

PubMed

Kõljalg, Siiri; Mändar, Rando; Sõber, Tiina; Rööp, Tiiu; Mändar, Reet

2017-06-01

While contamination of mobile phones in the hospital has been found to be common in several studies, little information about bacterial abundance on phones used in the community is available. Our aim was to quantitatively determine the bacterial contamination of secondary school students' mobile phones. Altogether 27 mobile phones were studied. The contact plate method and microbial identification using MALDI-TOF mass spectrometer were used for culture studies. Quantitative PCR reaction for detection of universal 16S rRNA, Enterococcus faecalis 16S rRNA and Escherichia coli allantoin permease were performed, and the presence of tetracycline ( tet A, tet B, tet M), erythromycin ( erm B) and sulphonamide ( sul 1) resistance genes was assessed. We found a high median bacterial count on secondary school students' mobile phones (10.5 CFU/cm 2 ) and a median of 17,032 bacterial 16S rRNA gene copies per phone. Potentially pathogenic microbes ( Staphylococcus aureus , Acinetobacter spp. , Pseudomonas spp., Bacillus cereus and Neisseria flavescens ) were found among dominant microbes more often on phones with higher percentage of E. faecalis in total bacterial 16S rRNA. No differences in contamination level or dominating bacterial species between phone owner's gender and between phone types (touch screen/keypad) were found. No antibiotic resistance genes were detected on mobile phone surfaces. Quantitative study methods revealed high level bacterial contamination of secondary school students' mobile phones.

High level bacterial contamination of secondary school students’ mobile phones

PubMed Central

Kõljalg, Siiri; Mändar, Rando; Sõber, Tiina; Rööp, Tiiu; Mändar, Reet

2017-01-01

Introduction While contamination of mobile phones in the hospital has been found to be common in several studies, little information about bacterial abundance on phones used in the community is available. Our aim was to quantitatively determine the bacterial contamination of secondary school students’ mobile phones. Methods Altogether 27 mobile phones were studied. The contact plate method and microbial identification using MALDI-TOF mass spectrometer were used for culture studies. Quantitative PCR reaction for detection of universal 16S rRNA, Enterococcus faecalis 16S rRNA and Escherichia coli allantoin permease were performed, and the presence of tetracycline (tetA, tetB, tetM), erythromycin (ermB) and sulphonamide (sul1) resistance genes was assessed. Results We found a high median bacterial count on secondary school students’ mobile phones (10.5 CFU/cm2) and a median of 17,032 bacterial 16S rRNA gene copies per phone. Potentially pathogenic microbes (Staphylococcus aureus, Acinetobacter spp., Pseudomonas spp., Bacillus cereus and Neisseria flavescens) were found among dominant microbes more often on phones with higher percentage of E. faecalis in total bacterial 16S rRNA. No differences in contamination level or dominating bacterial species between phone owner’s gender and between phone types (touch screen/keypad) were found. No antibiotic resistance genes were detected on mobile phone surfaces. Conclusion Quantitative study methods revealed high level bacterial contamination of secondary school students’ mobile phones. PMID:28626737

Antibacterial activity of antibacterial cutting boards in household kitchens.

PubMed

Kounosu, Masayuki; Kaneko, Seiichi

2007-12-01

We examined antibacterial cutting boards with antibacterial activity values of either "2" or "4" in compliance with the JIS Z 2801 standard, and compared their findings with those of cutting boards with no antibacterial activity. These cutting boards were used in ten different households, and we measured changes in the viable cell counts of several types of bacteria with the drop plate method. We also identified the detected bacterial flora and measured the minimum antimicrobial concentrations of several commonly used antibacterial agents against the kinds of bacteria identified to determine the expected antibacterial activity of the respective agents. Cutting boards with activity values of both "2" and "4" proved to be antibacterial in actual use, although no correlation between the viable cell counts and the antibacterial activity values was observed. In the kitchen environment, large quantities of Pseudomonas, Flavobacterium, Micrococcus, and Bacillus were detected, and it was confirmed that common antibacterial agents used in many antibacterial products are effective against these bacterial species. In addition, we measured the minimum antimicrobial concentrations of the agents against lactobacillus, a typical good bacterium, and discovered that this bacterium is less sensitive to these antibacterial agents compared to more common bacteria.

A comparative evaluation of antimicrobial efficacy and flow properties for Epiphany, Guttaflow and AH-Plus sealer.

PubMed

Nawal, R R; Parande, M; Sehgal, R; Naik, A; Rao, N R

2011-04-01

To test the antimicrobial efficacy and flow properties of Guttaflow, Epiphany sealer and AH-Plus sealer. With the use of Enterococcus faecalis ATCC 29212 as a test organism, both the agar diffusion test (ADT) and direct contact test (DCT) were performed. For DCT, sealers were mixed and placed over the bottom of sterile screw-capped test tubes. A 50 μL bacterial suspension was placed on the tested material samples. Bacteria were allowed to directly come in contact with the sealers for 1 h at 37 °C in one group and for 24 h in the other group. The suspensions were then diluted and inoculated over blood agar plates, and bacterial colony counts were determined with the use of a digital colony counter. The data in both 1- and 24-h groups were individually analysed using repeated measures ANOVA. Kruskal Wallis tests were further used to obtain comparison between 1- and 24-h results for all three sealers. In the flow assay, the sealers were placed between two glass slides, and a weight of 500 g was placed on the top of the glass. The diameters of the formed discs were recorded. For both the ADT and DCT tests, Epiphany and AH-Plus sealer reduced the bacterial counts significantly (P = 0.000). Epiphany produced a greater reduction in bacterial counts when compared to AH-Plus in both the tests (P = 0.000). Guttaflow paste failed to show any antibacterial activity in both ADT & DCT. According to the flow test, all root canal sealers flowed; Epiphany sealer had the maximum flow under the given conditions, followed by AH-Plus sealer and Guttaflow paste. Antimicrobial activity of the sealers was greatest for Epiphany followed by AH-Plus sealer and Guttaflow. Epiphany sealer had the maximum flow followed by AH-Plus sealer and Guttaflow. © 2010 International Endodontic Journal.

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.

Microbial air quality and bacterial surface contamination in ambulances during patient services.

PubMed

Luksamijarulkul, Pipat; Pipitsangjan, Sirikun

2015-03-01

We sought to assess microbial air quality and bacterial surface contamination on medical instruments and the surrounding areas among 30 ambulance runs during service. We performed a cross-sectional study of 106 air samples collected from 30 ambulances before patient services and 212 air samples collected during patient services to assess the bacterial and fungal counts at the two time points. Additionally, 226 surface swab samples were collected from medical instrument surfaces and the surrounding areas before and after ambulance runs. Groups or genus of isolated bacteria and fungi were preliminarily identified by Gram's stain and lactophenol cotton blue. Data were analyzed using descriptive statistics, t-test, and Pearson's correlation coefficient with a p-value of less than 0.050 considered significant. The mean and standard deviation of bacterial and fungal counts at the start of ambulance runs were 318±485cfu/m(3) and 522±581cfu/m(3), respectively. Bacterial counts during patient services were 468±607cfu/m(3) and fungal counts were 656±612cfu/m(3). Mean bacterial and fungal counts during patient services were significantly higher than those at the start of ambulance runs, p=0.005 and p=0.030, respectively. For surface contamination, the overall bacterial counts before and after patient services were 0.8±0.7cfu/cm(2) and 1.3±1.1cfu/cm(2), respectively (p<0.001). The predominant isolated bacteria and fungi were Staphylococcus spp. and Aspergillus spp., respectively. Additionally, there was a significantly positive correlation between bacterial (r=0.3, p<0.010) and fungal counts (r=0.2, p=0.020) in air samples and bacterial counts on medical instruments and allocated areas. This study revealed high microbial contamination (bacterial and fungal) in ambulance air during services and higher bacterial contamination on medical instrument surfaces and allocated areas after ambulance services compared to the start of ambulance runs. Additionally, bacterial and fungal counts in ambulance air showed a significantly positive correlation with the bacterial surface contamination on medical instruments and allocated areas. Further studies should be conducted to determine the optimal intervention to reduce microbial contamination in the ambulance environment.

A comparative study of biofilm formation by Shiga toxigenic Escherichia coli using epifluorescence microscopy on stainless steel and a microtitre plate method.

PubMed

Rivas, Lucia; Dykes, Gary A; Fegan, Narelle

2007-04-01

Attachment of Shiga toxigenic Escherichia coli (STEC) to surfaces and the formation of biofilms may enhance persistence in a food processing environment and present a risk of contaminating products. Seven strains of STEC and three non-STEC strains were selected to compare two biofilm quantification methods; epifluorescence microscopy on stainless steel (SS) and a microtitre plate assay. The influence of prior growth in planktonic (nutrient broth) and sessile (nutrient agar) culture on biofilm production, as well as expression of surface structures and the possession of antigen 43 (encoded by agn43) on biofilm formation were also investigated. Biofilms were produced in diluted nutrient broth at 25 degrees C for 24 and 48 h. Curli expression was determined using congo red indicator agar, while the presence of agn43 was determined using polymerase chain reaction. No correlation was found between counts for epifluorescence microscopy on SS and the absorbance values obtained with the microtitre plate method for planktonic and sessile grown cultures. Different abilities of individual STEC strains to attach to SS and microtitre plates were found with some strains attaching better to each surface following growth in either planktonic or sessile culture. All O157 STEC strains had low biofilm counts on SS for planktonic and sessile grown cultures; however, one STEC O157:H- strain (EC516) had significantly greater (p<0.05) biofilm production on microtitre plates compared to the other O157 STEC strains. EC516 and other STEC (O174:H21 and O91:H21) strains expressing curli fimbriae were found to produce significantly greater (p<0.05) biofilms on microtitre plates compared to the non-curli expressing strains. No relationship was found between the production of type-I fimbriae, motility, agn43 and bacterial physicochemical properties (previously determined) and biofilm formation on SS or microtitre plates. Variations between the two biofilm determination methods may suggest that the biofilm production on microtitre plates may not be appropriate to represent other surfaces such as SS and that caution should be taken when selecting a method to quantify biofilm production on a surface.

Comparison of fluorescence microscopy and solid-phase cytometry methods for counting bacteria in water

USGS Publications Warehouse

Lisle, John T.; Hamilton, Martin A.; Willse, Alan R.; McFeters, Gordon A.

2004-01-01

Total direct counts of bacterial abundance are central in assessing the biomass and bacteriological quality of water in ecological and industrial applications. Several factors have been identified that contribute to the variability in bacterial abundance counts when using fluorescent microscopy, the most significant of which is retaining an adequate number of cells per filter to ensure an acceptable level of statistical confidence in the resulting data. Previous studies that have assessed the components of total-direct-count methods that contribute to this variance have attempted to maintain a bacterial cell abundance value per filter of approximately 106 cells filter-1. In this study we have established the lower limit for the number of bacterial cells per filter at which the statistical reliability of the abundance estimate is no longer acceptable. Our results indicate that when the numbers of bacterial cells per filter were progressively reduced below 105, the microscopic methods increasingly overestimated the true bacterial abundance (range, 15.0 to 99.3%). The solid-phase cytometer only slightly overestimated the true bacterial abundances and was more consistent over the same range of bacterial abundances per filter (range, 8.9 to 12.5%). The solid-phase cytometer method for conducting total direct counts of bacteria was less biased and performed significantly better than any of the microscope methods. It was also found that microscopic count data from counting 5 fields on three separate filters were statistically equivalent to data from counting 20 fields on a single filter.

Bladder irrigation in patients with indwelling catheters.

PubMed

Bruun, J N; Digranes, A

1978-01-01

The effect of intermittent bladder irrigation on the bacterial counts in urine samples was studied in patients with indwelling catheter and pre-existing urinary tract infection. Four different irrigating solutions were used. Irrigation with saline or 0.25% acetic acid had no effect on the urinary bacterial count. The bacterial counts were effectively reduced during intermittent irrigation both with 0.02% chlorhexidine and with 0.25% silver nitrate. Silver nitrate gave the greatest reduction of bacterial counts but chlorhexidine is preferable due to fewer side effects and greater convenience.

A miniaturized counting technique for anaerobic bacteria.

PubMed

Sharpe, A N; Pettipher, G L; Lloyd, G R

1976-12-01

A miniaturized counting technique gave results as good as the pour-plate and Most Probable Number (MPN) techniques for enumeration of clostridia spp. and anaerobic isolates from the gut. Highest counts were obtained when ascorbic acid (1%) and dithiothreitol (0.015%) were added to the reinforced clostridial medium used for counting. This minimized the effect of exposure to air before incubation. The miniature technique allowed up to 40 samples to be plated and incubated in one McIntosh-Filde's-type anaerobic jar, compared with 3 or 4 by the normal pour plate.

Microbiological quality of raw milk attributable to prolonged refrigeration conditions.

PubMed

Vithanage, Nuwan R; Dissanayake, Muditha; Bolge, Greg; Palombo, Enzo A; Yeager, Thomas R; Datta, Nivedita

2017-02-01

Refrigerated storage of raw milk is a prerequisite in dairy industry. However, temperature abused conditions in the farming and processing environments can significantly affect the microbiological quality of raw milk. Thus, the present study investigated the effect of different refrigeration conditions such as 2, 4, 6, 8, 10 and 12 °C on microbiological quality of raw milk from three different dairy farms with significantly different initial microbial counts. The bacterial counts (BC), protease activity (PA), proteolysis (PL) and microbial diversity in raw milk were determined during storage. The effect of combined heating (75 ± 0·5 °C for 15 s) and refrigeration on controlling those contaminating microorganisms was also investigated. Results of the present study indicated that all of the samples showed increasing BC, PA and PL as a function of temperature, time and initial BC with a significant increase in those criteria ≥6 °C. Similar trends in BC, PA and PL were observed during the extended storage of raw milk at 4 °C. Both PA and PL showed strong correlation with the psychrotrophic proteolytic count (PPrBC: at ≥4 °C) and thermoduric psychrotrophic count (TDPC: at ≥8 °C) compared to total plate count (TPC) and psychrotrophic bacterial count (PBC), that are often used as the industry standard. Significant increases in PA and PL were observed when PPrBC and TDPC reached 5 × 104 cfu/ml and 1 × 104 cfu/ml, and were defined as storage life for quality (S LQ), and storage life for safety (S LS) aspects, respectively. The storage conditions also significantly affected the microbial diversity, where Pseudomonas fluorescens and Bacillus cereus were found to be the most predominant isolates. However, deep cooling (2 °C) and combination of heating and refrigeration (≤4 °C) significantly extended the S LQ and S Ls of raw milk.

Technical note: enumeration of mesophilic aerobes in milk: evaluation of standard official protocols and Petrifilm aerobic count plates.

PubMed

Freitas, R; Nero, L A; Carvalho, A F

2009-07-01

Enumeration of mesophilic aerobes (MA) is the main quality and hygiene parameter for raw and pasteurized milk. High levels of these microorganisms indicate poor conditions in production, storage, and processing of milk, and also the presence of pathogens. Fifteen raw and 15 pasteurized milk samples were submitted for MA enumeration by a conventional plating method (using plate count agar) and Petrifilm Aerobic Count plates (3M, St. Paul, MN), followed by incubation according to 3 official protocols: IDF/ISO (incubation at 30 degrees C for 72 h), American Public Health Association (32 degrees C for 48 h), and Brazilian Ministry of Agriculture (36 degrees C for 48 h). The results were compared by linear regression and ANOVA. Considering the results from conventional methodology, good correlation indices and absence of significant differences between mean counts were observed, independent of type of milk sample (raw or pasteurized) and incubation conditions (IDF/ISO, American Public Health Association, or Ministry of Agriculture). Considering the results from Petrifilm Aerobic Count plates, good correlation indices and absence of significant differences were only observed for raw milk samples. The microbiota of pasteurized milk interfered negatively with the performance of Petrifilm Aerobic Count plates, probably because of the presence of microorganisms that poorly reduce the dye indicator of this system.

Attachment of Asaia bogorensis Originating in Fruit-Flavored Water to Packaging Materials

PubMed Central

Otlewska, Anna; Antolak, Hubert

2014-01-01

The objective of this study was to investigate the adhesion of isolated spoilage bacteria to packaging materials used in the food industry. Microorganisms were isolated from commercial fruit-flavored mineral water in plastic bottles with flocks as a visual defect. The Gram-negative rods were identified using the molecular method through the amplification of a partial region of the 16S rRNA gene. Based on the sequence identity (99.6%) between the spoilage organism and a reference strain deposited in GenBank, the spoilage isolate was identified as Asaia bgorensis. Experiments on bacterial adhesion were conducted using plates made of glass and polystyrene (packaging materials commonly used in the beverage industry). Cell adhesion ability was determined using luminometry, plate count, and the microscopic method. The strain of A. bogorensis was characterized by strong adhesion properties which were dependent on the surface type, with the highest cell adhesion detected on polystyrene. PMID:25295262

Enumeration of total aerobic microorganisms in foods by SimPlate Total Plate Count-Color Indicator methods and conventional culture methods: collaborative study.

PubMed

Feldsine, Philip T; Leung, Stephanie C; Lienau, Andrew H; Mui, Linda A; Townsend, David E

2003-01-01

The relative efficacy of the SimPlate Total Plate Count-Color Indicator (TPC-CI) method (SimPlate 35 degrees C) was compared with the AOAC Official Method 966.23 (AOAC 35 degrees C) for enumeration of total aerobic microorganisms in foods. The SimPlate TPC-CI method, incubated at 30 degrees C (SimPlate 30 degrees C), was also compared with the International Organization for Standardization (ISO) 4833 method (ISO 30 degrees C). Six food types were analyzed: ground black pepper, flour, nut meats, frozen hamburger patties, frozen fruits, and fresh vegetables. All foods tested were naturally contaminated. Nineteen laboratories throughout North America and Europe participated in the study. Three method comparisons were conducted. In general, there was <0.3 mean log count difference in recovery among the SimPlate methods and their corresponding reference methods. Mean log counts between the 2 reference methods were also very similar. Repeatability (Sr) and reproducibility (SR) standard deviations were similar among the 3 method comparisons. The SimPlate method (35 degrees C) and the AOAC method were comparable for enumerating total aerobic microorganisms in foods. Similarly, the SimPlate method (30 degrees C) was comparable to the ISO method when samples were prepared and incubated according to the ISO method.

Enumeration procedure for monitoring test microbe populations on inoculated carriers in AOAC use-dilution methods.

PubMed

Tomasino, Stephen F; Fiumara, Rebecca M; Cottrill, Michele P

2006-01-01

The AOAC Use-Dilution methods do not provide procedures to enumerate the test microbe on stainless steel carriers (penicylinders) or guidance on the expected target populations of the test microbe (i.e., a performance standard). This report describes the procedures used by the U.S. Environmental Protection Agency to enumerate the test microbe (carrier counts) associated with conducting the Use-Dilution method with Staphylococcus aureus (Method 955.15) and Pseudomonas aeruginosa (Method 964.02) and the examination of historical data. The carrier count procedure involves the random selection of carriers, shearing bacterial cells from the carrier surface through sonication, and plating of serially diluted inoculum on trypticase soy agar. For each Use-Dilution test conducted, the official AOAC method was strictly followed for carrier preparation, culture initiation, test culture preparation, and carrier inoculation steps. Carrier count data from 78 Use-Dilution tests conducted over a 6-year period were compiled and analyzed. A mean carrier count of 6.6 logs (approximately 4.0 x 10(6) colony-forming units/carrier) was calculated for both S. aureus and P. aeruginosa. Of the mean values, 95% fell within +/- 2 repeatability standard deviations. The enumeration procedure and target carrier counts are desirable for standardizing the Use-Dilution methods, increasing their reproducibility, and ensuring the quality of the data.

Microbial Air Quality and Bacterial Surface Contamination in Ambulances During Patient Services

PubMed Central

Luksamijarulkul, Pipat; Pipitsangjan, Sirikun

2015-01-01

Objectives We sought to assess microbial air quality and bacterial surface contamination on medical instruments and the surrounding areas among 30 ambulance runs during service. Methods We performed a cross-sectional study of 106 air samples collected from 30 ambulances before patient services and 212 air samples collected during patient services to assess the bacterial and fungal counts at the two time points. Additionally, 226 surface swab samples were collected from medical instrument surfaces and the surrounding areas before and after ambulance runs. Groups or genus of isolated bacteria and fungi were preliminarily identified by Gram’s stain and lactophenol cotton blue. Data were analyzed using descriptive statistics, t-test, and Pearson’s correlation coefficient with a p-value of less than 0.050 considered significant. Results The mean and standard deviation of bacterial and fungal counts at the start of ambulance runs were 318±485cfu/m3 and 522±581cfu/m3, respectively. Bacterial counts during patient services were 468±607cfu/m3 and fungal counts were 656±612cfu/m3. Mean bacterial and fungal counts during patient services were significantly higher than those at the start of ambulance runs, p=0.005 and p=0.030, respectively. For surface contamination, the overall bacterial counts before and after patient services were 0.8±0.7cfu/cm2 and 1.3±1.1cfu/cm2, respectively (p<0.001). The predominant isolated bacteria and fungi were Staphylococcus spp. and Aspergillus spp., respectively. Additionally, there was a significantly positive correlation between bacterial (r=0.3, p<0.010) and fungal counts (r=0.2, p=0.020) in air samples and bacterial counts on medical instruments and allocated areas. Conclusions This study revealed high microbial contamination (bacterial and fungal) in ambulance air during services and higher bacterial contamination on medical instrument surfaces and allocated areas after ambulance services compared to the start of ambulance runs. Additionally, bacterial and fungal counts in ambulance air showed a significantly positive correlation with the bacterial surface contamination on medical instruments and allocated areas. Further studies should be conducted to determine the optimal intervention to reduce microbial contamination in the ambulance environment. PMID:25960835

The development and evaluation of ultrasound for the treatment of bacterial suspensions. A study of frequency, power and sonication time on cultured Bacillus species.

PubMed

Joyce, E; Phull, S S; Lorimer, J P; Mason, T J

2003-10-01

Some species of bacteria produce colonies and spores which agglomerate in spherical clusters (Bacillus subtilis) and this serves as a protection for the organisms inside against biocidal attack. Flocs of fine particles e.g. clay can entrap bacteria which can also protect them against the biocides. It is because of problems such as these that alternative methods of disinfecting water are under active investigation. One such method is the use of power ultrasound, either alone or in combination with other methods. Ultrasound is able to inactivate bacteria and deagglomerate bacterial clusters or flocs through a number of physical, mechanical and chemical effects arising from acoustic cavitation. The aim of this study was to investigate the effect of power ultrasound at different powers and frequencies on Bacillus subtilis. Viable plate count techniques were used as a measure of microbial activity. Results showed a significant increase in percent kill for Bacillus species with increasing duration of exposure and intensity of ultrasound in the low-kilohertz range (20 and 38 kHz). Results obtained at two higher frequencies (512 and 850 kHz) indicated a significant increase in bacteria count suggesting declumping. In assessing the bacterial kill with time under different sonication regimes three types of behaviour were characterized: High power ultrasound (lower frequencies) in low volumes of bacterial suspension results in a continuous reduction in bacterial cell numbers i.e. the kill rate predominates. High power ultrasound (lower frequencies) in larger volumes results in an initial rise in cell numbers suggesting declumping of the bacteria but this initial rise then falls as the declumping finishes and the kill rate becomes more important. Low intensity ultrasound (higher frequencies) gives an initial rise in cell numbers as a result of declumping. The kill rate is low and so there is no significant subsequent decrease in bacterial cell numbers.

Soil microbial response to waste potassium silicate drilling fluid.

PubMed

Yao, Linjun; Naeth, M Anne; Jobson, Allen

2015-03-01

Potassium silicate drilling fluids (PSDF) are a waste product of the oil and gas industry with potential for use in land reclamation. Few studies have examined the influence of PSDF on abundance and composition of soil bacteria and fungi. Soils from three representative locations for PSDF application in Alberta, Canada, with clay loam, loam and sand textures were studied with applications of unused, used once and used twice PSDF. For all three soils, applying ≥40 m3/ha of used PSDF significantly affected the existing soil microbial flora. No microbiota was detected in unused PSDF without soil. Adding used PSDF to soil significantly increased total fungal and aerobic bacterial colony forming units in dilution plate counts, and anaerobic denitrifying bacteria numbers in serial growth experiments. Used PSDF altered bacterial and fungal colony forming unit ratios of all three soils. Copyright © 2015. Published by Elsevier B.V.

Microbial Quality and Phylogenetic Diversity of Fresh Rainwater and Tropical Freshwater Reservoir

PubMed Central

Kaushik, Rajni; Balasubramanian, Rajasekhar; Dunstan, Hugh

2014-01-01

The impact of rainwater on the microbial quality of a tropical freshwater reservoir through atmospheric wet deposition of microorganisms was studied for the first time. Reservoir water samples were collected at four different sampling points and rainwater samples were collected in the immediate vicinity of the reservoir sites for a period of four months (January to April, 2012) during the Northeast monsoon period. Microbial quality of all fresh rainwater and reservoir water samples was assessed based on the counts for the microbial indicators: Escherichia coli (E. coli), total coliforms, and Enterococci along with total heterotrophic plate counts (HPC). The taxonomic richness and phylogenetic relationship of the freshwater reservoir with those of the fresh rainwater were also assessed using 16 S rRNA gene clone library construction. The levels of E. coli were found to be in the range of 0 CFU/100 mL – 75 CFU/100 mL for the rainwater, and were 10–94 CFU/100 mL for the reservoir water. The sampling sites that were influenced by highway traffic emissions showed the maximum counts for all the bacterial indicators assessed. There was no significant increase in the bacterial abundances observed in the reservoir water immediately following rainfall. However, the composite fresh rainwater and reservoir water samples exhibited broad phylogenetic diversity, including sequences representing Betaproteobacteria, Alphaproteobacteria, Gammaproteobacteria, Actinobacteria, Lentisphaerae and Bacteriodetes. Members of the Betaproteobacteria group were the most dominant in both fresh rainwater and reservoir water, followed by Alphaproteobacteria, Sphingobacteria, Actinobacteria and Gammaproteobacteria. PMID:24979573

Microbial quality and phylogenetic diversity of fresh rainwater and tropical freshwater reservoir.

PubMed

Kaushik, Rajni; Balasubramanian, Rajasekhar; Dunstan, Hugh

2014-01-01

The impact of rainwater on the microbial quality of a tropical freshwater reservoir through atmospheric wet deposition of microorganisms was studied for the first time. Reservoir water samples were collected at four different sampling points and rainwater samples were collected in the immediate vicinity of the reservoir sites for a period of four months (January to April, 2012) during the Northeast monsoon period. Microbial quality of all fresh rainwater and reservoir water samples was assessed based on the counts for the microbial indicators: Escherichia coli (E. coli), total coliforms, and Enterococci along with total heterotrophic plate counts (HPC). The taxonomic richness and phylogenetic relationship of the freshwater reservoir with those of the fresh rainwater were also assessed using 16 S rRNA gene clone library construction. The levels of E. coli were found to be in the range of 0 CFU/100 mL-75 CFU/100 mL for the rainwater, and were 10-94 CFU/100 mL for the reservoir water. The sampling sites that were influenced by highway traffic emissions showed the maximum counts for all the bacterial indicators assessed. There was no significant increase in the bacterial abundances observed in the reservoir water immediately following rainfall. However, the composite fresh rainwater and reservoir water samples exhibited broad phylogenetic diversity, including sequences representing Betaproteobacteria, Alphaproteobacteria, Gammaproteobacteria, Actinobacteria, Lentisphaerae and Bacteriodetes. Members of the Betaproteobacteria group were the most dominant in both fresh rainwater and reservoir water, followed by Alphaproteobacteria, Sphingobacteria, Actinobacteria and Gammaproteobacteria.

Inactivation of Selected Bacterial Pathogens in Dairy Cattle Manure by Mesophilic Anaerobic Digestion (Balloon Type Digester)

PubMed Central

Manyi-Loh, Christy E.; Mamphweli, Sampson N.; Meyer, Edson L.; Okoh, Anthony I.; Makaka, Golden; Simon, Michael

2014-01-01

Anaerobic digestion of animal manure in biogas digesters has shown promise as a technology in reducing the microbial load to safe and recommended levels. We sought to treat dairy manure obtained from the Fort Hare Dairy Farm by investigating the survival rates of bacterial pathogens, through a total viable plate count method, before, during and after mesophilic anaerobic digestion. Different microbiological media were inoculated with different serial dilutions of manure samples that were withdrawn from the biogas digester at 3, 7 and 14 day intervals to determine the viable cells. Data obtained indicated that the pathogens of public health importance were 90%–99% reduced in the order: Campylobacter sp. (18 days) < Escherichia coli sp. (62 days) < Salmonella sp. (133 days) from a viable count of 10.1 × 103, 3.6 × 105, 7.4 × 103 to concentrations below the detection limit (DL = 102 cfu/g manure), respectively. This disparity in survival rates may be influenced by the inherent characteristics of these bacteria, available nutrients as well as the stages of the anaerobic digestion process. In addition, the highest p-value i.e., 0.957 for E. coli showed the statistical significance of its model and the strongest correlation between its reductions with days of digestion. In conclusion, the results demonstrated that the specific bacterial pathogens in manure can be considerably reduced through anaerobic digestion after 133 days. PMID:25026086

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.

Bacteriological quality of the wastewater used for irrigation at the vegetable farms in Korle-bu Teaching Hospital, Accra Metropolis, Ghana.

PubMed

Pesewu, George A; Bentum, Daniel; Olu-Taiwo, Michael A; Glover, Kathreen K; Yirenya-Tawiah, Dzidzo R

2017-01-01

Many developing countries, including Ghana, are water stressed. As such, farmers, particularly those in urban areas, have adopted the use of wastewater for irrigation. This study evaluated the bacteriological water quality of the wastewater used for irrigation in the vegetable farms at Korle-Bu Teaching Hospital (KBTH), Accra Metropolis, Ghana. In all, 40 wastewater samples were collected and analysed bacteriologically using the total aerobic plate count method. The isolated bacteria were identified biochemically using Bergey's manual for determinative bacteriology. Mean total bacterial colony count values in the range of 2.75-4.44 × 10 5 CFU/100 mL were isolated which far exceeds values of 1 × 10 3 /100 mL recommended by the World Health Organization (WHO) for unrestricted irrigation of crops likely to be eaten raw. Enterobacter cloacae (51.4%), Klebsiella sp. (24.1%), Pseudomonas aeruginosa (11.3%), Salmonella typhi (10.6%), Escherichia coli (2.2%) and Proteus sp. (0.4%) were the predominant bacteria isolated. Growers should use treated wastewater for farming while processors and consumers should minimize contamination risks of produce from the vegetable farms/garden to the plate. © The Author(s) 2016.

Aerobic Heterotrophic Bacterial Populations of Sewage and Activated Sludge

PubMed Central

Prakasam, T. B. S.; Dondero, N. C.

1970-01-01

An activated sludge from a sewage treatment plant and a laboratory activated sludge developed on an artificial waste were compared for their ability to utilize 11 aromatic compounds. There were several significant differences between them. The laboratory sludge contained higher numbers of organisms and metabolized the aromatics to a greater extent. Laboratory activated sludges acclimated to utilization of the aromatics differed from each other in population structure and the pattern of oxygen consumption with aromatic substrates. The oxidative patterns of uncontrolled mixed populations were unreliable for investigating metabolic pathways. Extracts of the various sludges elevated the plate counts of the sludges. PMID:5418946

Comparison of Standard Culture-Based Method to Culture-Independent Method for Evaluation of Hygiene Effects on the Hand Microbiome.

PubMed

Zapka, C; Leff, J; Henley, J; Tittl, J; De Nardo, E; Butler, M; Griggs, R; Fierer, N; Edmonds-Wilson, S

2017-03-28

Hands play a critical role in the transmission of microbiota on one's own body, between individuals, and on environmental surfaces. Effectively measuring the composition of the hand microbiome is important to hand hygiene science, which has implications for human health. Hand hygiene products are evaluated using standard culture-based methods, but standard test methods for culture-independent microbiome characterization are lacking. We sampled the hands of 50 participants using swab-based and glove-based methods prior to and following four hand hygiene treatments (using a nonantimicrobial hand wash, alcohol-based hand sanitizer [ABHS], a 70% ethanol solution, or tap water). We compared results among culture plate counts, 16S rRNA gene sequencing of DNA extracted directly from hands, and sequencing of DNA extracted from culture plates. Glove-based sampling yielded higher numbers of unique operational taxonomic units (OTUs) but had less diversity in bacterial community composition than swab-based sampling. We detected treatment-induced changes in diversity only by using swab-based samples ( P < 0.001); we were unable to detect changes with glove-based samples. Bacterial cell counts significantly decreased with use of the ABHS ( P < 0.05) and ethanol control ( P < 0.05). Skin hydration at baseline correlated with bacterial abundances, bacterial community composition, pH, and redness across subjects. The importance of the method choice was substantial. These findings are important to ensure improvement of hand hygiene industry methods and for future hand microbiome studies. On the basis of our results and previously published studies, we propose recommendations for best practices in hand microbiome research. IMPORTANCE The hand microbiome is a critical area of research for diverse fields, such as public health and forensics. The suitability of culture-independent methods for assessing effects of hygiene products on microbiota has not been demonstrated. This is the first controlled laboratory clinical hand study to have compared traditional hand hygiene test methods with newer culture-independent characterization methods typically used by skin microbiologists. This study resulted in recommendations for hand hygiene product testing, development of methods, and future hand skin microbiome research. It also demonstrated the importance of inclusion of skin physiological metadata in skin microbiome research, which is atypical for skin microbiome studies. Copyright © 2017 Zapka et al.

Characterization of bacterial communities and functions of two submerged soils from San Vitale park (Italy)

NASA Astrophysics Data System (ADS)

Mocali, Stefano; Chiellini, Carolina; Lagomarsino, Alessandra; Ferronato, Chiara; Vittori Antisari, Livia; Vianello, Gilmo

2015-04-01

Subaqueous soils has been introduced in the last edition of the Keys to Soil Taxonomy (Soil surveystaff, 2014), to describe soils covered by a water column of up to 2.5 m where different pedogenetic processes can be recognized. However, the role of bacterial community structure and function in such environments and its potential use as pedogenetic indicator is still largely unknown. Two submerged soils (WAS-2 and WAS-4) were collected from San Vitale park (Italy), a site where the evolution of the landscape from subaqueous wetland to interdunal and dunal system, and the interfacing of freshwater with saltwater, made this site particularly suitable for examining the pedogenetic indicators which can characterize and predict the soil hydromorphism in trasitional ecosystems. The two soils were classified and their physicochemical and morphological features were investigated. Selective media were used to isolate both culturable aerobic and anaerobic (microaerophilic) bacteria associated with each horizon. In WAS-2 seven horizons were identified (depths 4-0, 0-6, 6-13, 13-20, 20-36, 36-59/60, and 59/60-83 cm) while in WAS-4, five horizons were identified (depths 0-14, 14-20, 20-40, 40-45, 45-100 cm) for a total of 12 horizons (samples). For each sample, aerobic bacterial plate count was performed on solid LB medium, coupled with microaerophilic bacterial plate count either on SA500 minimal medium and AYE medium (0.5% soft agar each). Molecular identification (16S rRNA gene sequencing) of ~100 strains isolated from each of the three used medium was performed, for a total of ~300 strains for each sample. To complete the characterization of the microbial communities in all horizons, Next Generation Sequencing (NGS) analysis was carried out with 454 platform on each of the 12 samples. Moreover, the N2O and CH4 emissions were determined from each pedon. All the parameters were used to highlight the similarities and the differences between and within the pedons. The results will potentially help to highlight some features related to the origin and morphogenesis of submerged soils, and also to clarify the process of differentiation of soil horizons from a biological point of view.

Demonstration of nitric oxide synthase activity in crustacean hemocytes and anti-microbial activity of hemocyte-derived nitric oxide.

PubMed

Yeh, Feng-Ching; Wu, Su-Hua; Lai, Chi-Yung; Lee, Chi-Ying

2006-05-01

We determined the biochemical characteristics of nitric oxide synthase (NOS) in hemocytes of the crayfish Procambarus clarkii and investigated the roles of hemocyte-derived NO in host defense. Biochemical analysis indicated the presence of a Ca2+ -independent NOS activity, which was elevated by lipopolysaccharide (LPS) treatment. When bacteria (Staphylococcus aureus) and hemocytes were co-incubated, adhesion of bacteria to hemocytes was observed. NO donor sodium nitroprusside (SNP) significantly increased the numbers of hemocytes to which bacteria adhered. Similarly, LPS elicited bacterial adhesion and the LPS-induced adhesion was prevented by NOS inhibitor NG-monomethyl-L-arginine (L-NMMA). Finally, plate count assay demonstrated that addition of LPS to the hemocytes/bacteria co-incubation resulted in a significant decrease in bacterial colony forming unit (CFU), and that L-NMMA reversed the decreasing effect of LPS on CFU. The combined results demonstrate the presence of a Ca2+ -independent LPS-inducible NOS activity in crayfish hemocytes and suggest that hemocyte-derived NO is involved in promoting bacterial adhesion to hemocytes and enhancing bactericidal activity of hemocytes.

Ultraviolet micro-Raman spectrograph for the detection of small numbers of bacterial cells

NASA Astrophysics Data System (ADS)

Chadha, S.; Nelson, W. H.; Sperry, J. F.

1993-11-01

The construction of a practical UV micro-Raman spectrograph capable of selective excitation of bacterial cells and other microscopic samples has been described. A reflective objective is used to focus cw laser light on a sample and at the same time collect the scattered light at 180°. With the aid of a quartz lens the image produced is focused on the slits of a spectrograph equipped with a single 2400 grooves/mm grating optimized for 250 nm. Spectra were detected by means of a blue-intensified diode array detector. Resonance Raman spectra of Bacillus subtilis and Flavobacterium capsulatum excited by the 257.2 nm output of a cw laser were recorded in the 900-1800 cm-1 region. Bacterial cells were immobilized on a quartz plate by means of polylysine and were counted visually. Cooling was required to retard sample degradation. Sample sizes ranged from 1 to 50 cells with excitation times varying from 15 to 180 s. Excellent spectra have been obtained from 20 cells in 15 s using a spectrograph having only 3% throughput.

Microbial structural diversity estimated by dilution-extinction of phenotypic traits and T-RFLP analysis along a land-use intensification gradient

NASA Technical Reports Server (NTRS)

Gomez, Elena del V.; Garland, Jay L.; Roberts, Michael S.

2004-01-01

The present work tested whether the relationship between functional traits and inoculum density reflected structural diversity in bacterial communities from a land-use intensification gradient applying a mathematical model. Terminal restriction fragment length polymorphism (T-RFLP) analysis was also performed to provide an independent assessment of species richness. Successive 10-fold dilutions of a soil suspension were inoculated onto Biolog GN(R) microplates. Soil bacterial density was determined by total cell and plate counts. The relationship between phenotypic traits and inoculum density fit the model, allowing the estimation of maximal phenotypic potential (Rmax) and inoculum density (KI) at which Rmax will be half-reduced. Though Rmax decreased with time elapsed since clearing of native vegetation, KI remained high in two of the disturbed sites. The genetic pool of bacterial community did not experience a significant reduction, but the active fraction responding in the Biolog assay was adversely affected, suggesting a reduction in the functional potential. c2004 Federation of European Microbiological Societies. Published by Elsevier B.V. All rights reserved.

Aseptic laboratory techniques: plating methods.

PubMed

Sanders, Erin R

2012-05-11

Microorganisms are present on all inanimate surfaces creating ubiquitous sources of possible contamination in the laboratory. Experimental success relies on the ability of a scientist to sterilize work surfaces and equipment as well as prevent contact of sterile instruments and solutions with non-sterile surfaces. Here we present the steps for several plating methods routinely used in the laboratory to isolate, propagate, or enumerate microorganisms such as bacteria and phage. All five methods incorporate aseptic technique, or procedures that maintain the sterility of experimental materials. Procedures described include (1) streak-plating bacterial cultures to isolate single colonies, (2) pour-plating and (3) spread-plating to enumerate viable bacterial colonies, (4) soft agar overlays to isolate phage and enumerate plaques, and (5) replica-plating to transfer cells from one plate to another in an identical spatial pattern. These procedures can be performed at the laboratory bench, provided they involve non-pathogenic strains of microorganisms (Biosafety Level 1, BSL-1). If working with BSL-2 organisms, then these manipulations must take place in a biosafety cabinet. Consult the most current edition of the Biosafety in Microbiological and Biomedical Laboratories (BMBL) as well as Material Safety Data Sheets (MSDS) for Infectious Substances to determine the biohazard classification as well as the safety precautions and containment facilities required for the microorganism in question. Bacterial strains and phage stocks can be obtained from research investigators, companies, and collections maintained by particular organizations such as the American Type Culture Collection (ATCC). It is recommended that non-pathogenic strains be used when learning the various plating methods. By following the procedures described in this protocol, students should be able to: Perform plating procedures without contaminating media. Isolate single bacterial colonies by the streak-plating method. Use pour-plating and spread-plating methods to determine the concentration of bacteria. Perform soft agar overlays when working with phage. Transfer bacterial cells from one plate to another using the replica-plating procedure. Given an experimental task, select the appropriate plating method.

Phylogenetic assessment of heterotrophic bacteria from a water distribution system using 16S rDNA sequencing.

PubMed

Tokajian, Sima T; Hashwa, Fuad A; Hancock, Ian C; Zalloua, Pierre A

2005-04-01

Determination of a heterotrophic plate count (HPC) for drinking-water samples alone is not enough to assess possible health hazards associated with sudden changes in the bacterial count. Speciation is very crucial to determine whether the population includes pathogens and (or) opportunistic pathogens. Most of the isolates recovered from drinking water samples could not be allocated to a specific phylogenetic branch based on the use of conventional diagnostic methods. The present study had to use phylogenetic analysis, which was simplified by determining and using the first 500-bp sequence of the 16S rDNA, to successfully identify the type and species of bacteria found in the samples. Gram-positive bacteria alpha-, beta-, and gamma-Proteobacteria were found to be the major groups representing the heterotrophic bacteria in drinking water. The study also revealed that the presence of sphingomonads in drinking water supplies may be much more common than has been reported so far and thus further studies are merited. The intermittent mode of supply, mainly characterized by water stagnation and flow interruption associated possibly with biofilm detachment, raised the possibility that the studied bacterial populations in such systems represented organisms coming from 2 different niches, the biofilm and the water column.

Biofilm responses to ageing and to a high phosphate load in a bench-scale drinking water system.

PubMed

Batté, Magali; Koudjonou, Boniface; Laurent, Patrick; Mathieu, Laurence; Coallier, Josée; Prévost, Michèle

2003-03-01

The effects of ageing and of phosphate load on drinking water biofilms developed on a polycarbonate substratum in the pseudo-equilibrium state have been evaluated. Phosphate was added in an amount higher than the stochiometric nutrient requirements of bacteria, at concentrations commonly applied in a drinking water distribution system for corrosion control. Multiple parameters were monitored: heterotrophic plate counts (HPCs), total direct counts (TDCs) and potential exoproteolytic activity (PEPA) in order to characterise changes in bacterial biofilms. The total carbohydrate, amino acid and phosphate contents of biofilms were analysed to characterise and monitor the biochemical composition of the biofilm.The three enumeration methods showed that a pseudo-equilibrium state was reached after 7 weeks of colonisation after which, the bacterial growth rate in the biofilm was 0.1 log per week on average. Bulk phosphate addition doubled the phosphate in the biofilm, but did not affect the other biological, physiological or chemical parameters measured. Polysaccharides increased in the biofilm with ageing and the dynamics of individual carbohydrate synthesis also varied with the age of the biofilm. Once pseudo-equilibrium, it was found that the total proteins were globally constant, whereas the spectra of some individual amino acids of the proteins had significantly changed.

A Molecular MST Approach to Investigate Fecal Indicator Bacteria in Bioaerosols, Bathing Water, Seaweed Wrack, and Sand at Recreational Beaches

NASA Astrophysics Data System (ADS)

Thoren, K. M.; Sinigalliano, C. D.

2016-02-01

Despite numerous cases of beach bacteria affecting millions of people worldwide, the persistence of the bacteria populations in coastal areas is still not well understood. The purpose of this study was to test the levels of persistence of Fecal Indicating Bacteria (FIB) of enterococci, Escherichia coli, and Human-source Bacteroidales, within the intertidal "swash zone" and the deeper waist zone in which people commonly bathe and play. In addition, the study sought to determine if these bacterial contaminants may also be found in aerosols at the beach. Measuring solar insolation in relation to bacterial persistence in seaweed wrack was used to determine if sunlight plays a role in modifying concentrations of FIB at the beach. Light intensity measured by a solar photometer and air quality measured by aerosol plate counts and qPCR Microbial Source Tracking (MST) was compared to varying locations where the beach samples were collected. Results from water samples demonstrate that bacteria measured using plate counts and qPCR were indeed higher within the swash zone than in the waist zone. This is in contrast with the way that the EPA currently measures and determines the public safety of beach waters. They commonly measure the waist zone, but disregard the swash zone. Results from beach bio-aerosol samples showed a wide variety of fungi and bacteria in the beach air, and qPCR MST analysis of these bio-aerosols showed the presence of FIBs such as enterococci on several of the aerosol collection plates. This emphasizes the need to collect samples from the entire beach instead of just measuring at an isolated area, and that exposure to microbial contaminants may include bathing water, beach sand, seaweed wrack, and bio-aerosols. Thus, the data reveals a potential way to identify harmful levels of bacteria and dangerous levels of poor air quality at recreational beaches. These results expound the need for broader assessment of potential beach contamination, not only the swimming water, but also the beach air, shoreline, and also varying depths of water, which can be extremely beneficial to reduce people's risk from microbial contamination exposure.

RELATIONSHIPS BETWEEN LEVELS OF HETEROTROPHIC BACTERIA AND WATER QUALITY PARAMETERS IN A DRINKING WATER DISTRIBUTION SYSTEM

EPA Science Inventory

Conventional plating methods were used to quantify heterotrophic bacteria from a drinking water distribution system. Three media, plate count agar (PCA), R2A agar and sheep blood agar (TSA-SB) were used to determine heterotrophic plate count (HPC) levels. Grab samples were collec...

Estimation method for serial dilution experiments.

PubMed

Ben-David, Avishai; Davidson, Charles E

2014-12-01

Titration of microorganisms in infectious or environmental samples is a corner stone of quantitative microbiology. A simple method is presented to estimate the microbial counts obtained with the serial dilution technique for microorganisms that can grow on bacteriological media and develop into a colony. The number (concentration) of viable microbial organisms is estimated from a single dilution plate (assay) without a need for replicate plates. Our method selects the best agar plate with which to estimate the microbial counts, and takes into account the colony size and plate area that both contribute to the likelihood of miscounting the number of colonies on a plate. The estimate of the optimal count given by our method can be used to narrow the search for the best (optimal) dilution plate and saves time. The required inputs are the plate size, the microbial colony size, and the serial dilution factors. The proposed approach shows relative accuracy well within ±0.1log10 from data produced by computer simulations. The method maintains this accuracy even in the presence of dilution errors of up to 10% (for both the aliquot and diluent volumes), microbial counts between 10(4) and 10(12) colony-forming units, dilution ratios from 2 to 100, and plate size to colony size ratios between 6.25 to 200. Published by Elsevier B.V.

Correlation between the neutrophil-lymphocyte count ratio and bacterial infection in patient with human immunodeficiency virus

NASA Astrophysics Data System (ADS)

Kusnadi, D.; Liwang, M. N. I.; Katu, S.; Mubin, A. H.; Halim, R.

2018-03-01

Parameters for starting antibiotic therapy such as CRP andleukocytosis are considered non-specific. Previous studies have shown the Neutrophil-Lymphocyte Count Ratio (NLCR) can serve as the basis of bacterial infection, the level of infection, and the basis of antibiotic therapy. Compared with the Procalcitonin parameter, this NLCR is rapid, an inexpensive and requires no additional sampling. To determine the correlation between The Neutrophil-LymphocyteCount Ratio to bacterial infection in HIV patients. This study was a cross-sectional observational approach to HIV subject at Wahidin Sudirohusodo and Hasanuddin University Hospital. The subjects performed routine blood, microbiology test,and blood Procalcitonin levels tests. Then performed NLCR calculations based on routine blood results. The subjects then grouped the presence or absence of bacterial infection.In 146 study subjects, there were 78 (53.4%) with bacterial infections and 68 (46.6%) without bacterial infection as controls. Subjects with bacterial infections had higher total neutrophils (84.83) compared with non-bacterial infections. Subjects with bacterial infections had total lymphocytes with an average of 8.51 lower than non-bacterial infections. Subjects with bacterial infections had higher NLCR values with an average of 12.80. The Neutrophil-Lymphocyte Count Ratio can become a marker of bacterial infection in HIV patients.

Performance Equivalence and Validation of the Soleris Automated System for Quantitative Microbial Content Testing Using Pure Suspension Cultures.

PubMed

Limberg, Brian J; Johnstone, Kevin; Filloon, Thomas; Catrenich, Carl

2016-09-01

Using United States Pharmacopeia-National Formulary (USP-NF) general method <1223> guidance, the Soleris(®) automated system and reagents (Nonfermenting Total Viable Count for bacteria and Direct Yeast and Mold for yeast and mold) were validated, using a performance equivalence approach, as an alternative to plate counting for total microbial content analysis using five representative microbes: Staphylococcus aureus, Bacillus subtilis, Pseudomonas aeruginosa, Candida albicans, and Aspergillus brasiliensis. Detection times (DTs) in the alternative automated system were linearly correlated to CFU/sample (R(2) = 0.94-0.97) with ≥70% accuracy per USP General Chapter <1223> guidance. The LOD and LOQ of the automated system were statistically similar to the traditional plate count method. This system was significantly more precise than plate counting (RSD 1.2-2.9% for DT, 7.8-40.6% for plate counts), was statistically comparable to plate counting with respect to variations in analyst, vial lots, and instruments, and was robust when variations in the operating detection thresholds (dTs; ±2 units) were used. The automated system produced accurate results, was more precise and less labor-intensive, and met or exceeded criteria for a valid alternative quantitative method, consistent with USP-NF general method <1223> guidance.

Advantageous Direct Quantification of Viable Closely Related Probiotics in Petit-Suisse Cheeses under In Vitro Gastrointestinal Conditions by Propidium Monoazide - qPCR

PubMed Central

Villarreal, Martha Lissete Morales; Padilha, Marina; Vieira, Antonio Diogo Silva; Franco, Bernadette Dora Gombossy de Melo; Martinez, Rafael Chacon Ruiz; Saad, Susana Marta Isay

2013-01-01

Species-specific Quantitative Real Time PCR (qPCR) alone and combined with the use of propidium monoazide (PMA) were used along with the plate count method to evaluate the survival of the probiotic strains Lactobacillus acidophilus La-5 and Bifidobacterium animalis subsp. lactis Bb-12, and the bacteriocinogenic and potentially probiotic strain Lactobacillus sakei subsp. sakei 2a in synbiotic (F1) and probiotic (F2) petit-suisse cheeses exposed throughout shelf-life to in vitro simulated gastrointestinal tract conditions. The three strains studied showed a reduction in their viability after the 6 h assay. Bb-12 displayed the highest survival capacity, above 72.6 and 74.6% of the initial populations, respectively, by plate count and PMA-qPCR, maintaining population levels in the range or above 6 log CFU/g. The prebiotic mix of inulin and FOS did not offer any additional protection for the strains against the simulated gastrointestinal environment. The microorganisms' populations were comparable among the three methods at the initial time of the assay, confirming the presence of mainly viable and culturable cells. However, with the intensification of the stress induced throughout the various stages of the in vitro test, the differences among the methods increased. The qPCR was not a reliable enumeration method for the quantification of intact bacterial populations, mixed with large numbers of injured and dead bacteria, as confirmed by the scanning electron microscopy results. Furthermore, bacteria plate counts were much lower (P<0.05) than with the PMA-qPCR method, suggesting the accumulation of stressed or dead microorganisms unable to form colonies. The use of PMA overcame the qPCR inability to differentiate between dead and alive cells. The combination of PMA and species-specific qPCR in this study allowed a quick and unequivocal way of enumeration of viable closely related species incorporated into probiotic and synbiotic petit-suisse cheeses and under stress conditions. PMID:24358142

Comparison of Dry Medium Culture Plates for Mesophilic Aerobic Bacteria in Milk, Ice Cream, Ham, and Codfish Fillet Products

PubMed Central

Park, Junghyun; Kim, Myunghee

2013-01-01

This study was performed to compare the performance of Sanita-Kun dry medium culture plate with those of traditional culture medium and Petrifilm dry medium culture plate for the enumeration of the mesophilic aerobic bacteria in milk, ice cream, ham, and codfish fillet. Mesophilic aerobic bacteria were comparatively evaluated in milk, ice cream, ham, and codfish fillet using Sanita-Kun aerobic count (SAC), Petrifilm aerobic count (PAC), and traditional plate count agar (PCA) media. According to the results, all methods showed high correlations of 0.989~1.000 and no significant differences were observed for enumerating the mesophilic aerobic bacteria in the tested food products. SAC method was easier to perform and count colonies efficiently as compared to the PCA and PAC methods. Therefore, we concluded that the SAC method offers an acceptable alternative to the PCA and PAC methods for counting the mesophilic aerobic bacteria in milk, ice cream, ham, and codfish fillet products. PMID:24551829

Conclusions on measurement uncertainty in microbiology.

PubMed

Forster, Lynne I

2009-01-01

Since its first issue in 1999, testing laboratories wishing to comply with all the requirements of ISO/IEC 17025 have been collecting data for estimating uncertainty of measurement for quantitative determinations. In the microbiological field of testing, some debate has arisen as to whether uncertainty needs to be estimated for each method performed in the laboratory for each type of sample matrix tested. Queries also arise concerning the estimation of uncertainty when plate/membrane filter colony counts are below recommended method counting range limits. A selection of water samples (with low to high contamination) was tested in replicate with the associated uncertainty of measurement being estimated from the analytical results obtained. The analyses performed on the water samples included total coliforms, fecal coliforms, fecal streptococci by membrane filtration, and heterotrophic plate counts by the pour plate technique. For those samples where plate/membrane filter colony counts were > or =20, uncertainty estimates at a 95% confidence level were very similar for the methods, being estimated as 0.13, 0.14, 0.14, and 0.12, respectively. For those samples where plate/membrane filter colony counts were <20, estimated uncertainty values for each sample showed close agreement with published confidence limits established using a Poisson distribution approach.

New procedure to reduce the time and cost of broncho-pulmonary specimen management using the Previ Isola® automated inoculation system.

PubMed

Nebbad-Lechani, Biba; Emirian, Aurélie; Maillebuau, Fabienne; Mahjoub, Nadia; Fihman, Vincent; Legrand, Patrick; Decousser, Jean-Winoc

2013-12-01

The microbiological diagnosis of respiratory tract infections requires serial manual dilutions of the clinical specimen before agar plate inoculation, disrupting the workflow in bacteriology clinical laboratories. Automated plating instrument systems have been designed to increase the speed, reproducibility and safety of this inoculating step; nevertheless, data concerning respiratory specimens are lacking. We tested a specific procedure that uses the Previ Isola® (bioMérieux, Craponne, France) to inoculate with broncho-pulmonary specimens (BPS). A total of 350 BPS from a university-affiliated hospital were managed in parallel using the manual reference and the automated methods (expectoration: 75; broncho-alveolar lavage: 68; tracheal aspiration: 17; protected distal sample: 190). A specific enumeration reading grid, a pre-liquefaction step and a fluidity test, performed before the inoculation, were designed for the automated method. The qualitative (i.e., the number of specimens yielding a bacterial count greater than the clinical threshold) and quantitative (i.e., the discrepancy within a 0.5 log value) concordances were 100% and 98.2%, respectively. The slimmest subgroup of expectorations could not be managed by the automated method (8%, 6/75). The technical time and cost savings (i.e., number of consumed plates) reached 50%. Additional studies are required for specific populations, such as cystic fibrosis specimens and associated bacterial variants. An automated decapper should be implemented to increase the biosafety of the process. The PREVI Isola® adapted procedure is a time- and cost-saving method for broncho-pulmonary specimen processing. © 2013.

Flow cytometry for immediate follow-up of drinking water networks after maintenance.

PubMed

Van Nevel, Sam; Buysschaert, Benjamin; De Roy, Karen; De Gusseme, Bart; Clement, Lieven; Boon, Nico

2017-03-15

Drinking water networks need maintenance every once in a while, either planned interventions or emergency repairs. When this involves opening of the water pipes, precautionary measures need to be taken to avoid contamination of the drinking water at all time. Drinking water suppliers routinely apply plating for faecal indicator organisms as quality control in such a situation. However, this takes at least 21 h of waiting time, which can be crucial when dealing with major supply pipes. A combination of flow cytometric (FCM) bacterial cell counts with FCM fingerprinting techniques is proposed in this study as a fast and sensitive additional technique. In three full scale situations, major supply pipes with 400-1050 mm diameter were emptied for maintenance, shock-chlorinated and flushed with large amounts of clean drinking water before taking back in operation. FCM measurements of the discharged flushing water revealed fast lowering and stabilizing bacterial concentrations once flushing is initiated. Immediate comparison with clean reference drinking water used for flushing was done, and the moment when both waters had similar bacterial concentrations was considered as the endpoint of the necessary flushing works. This was usually after 2-4 h of flushing. FCM fingerprinting, based on both bacteria and FCM background, was used as additional method to verify how similar flushing and reference samples were and yielded similar results. The FCM approved samples were several hours later approved as well by the drinking water supplier after plating and incubation for total Coliforms and Enterococci. These were used as decisive control to set the pipes back in operation. FCM proved to be a more conservative test than plating, yet it yielded immediate results. Application of these FCM methods can therefore avoid long unnecessary waiting times and large drinking water losses. Copyright © 2016 Elsevier Ltd. All rights reserved.

Use of simulation tools to illustrate the effect of data management practices for low and negative plate counts on the estimated parameters of microbial reduction models.

PubMed

Garcés-Vega, Francisco; Marks, Bradley P

2014-08-01

In the last 20 years, the use of microbial reduction models has expanded significantly, including inactivation (linear and nonlinear), survival, and transfer models. However, a major constraint for model development is the impossibility to directly quantify the number of viable microorganisms below the limit of detection (LOD) for a given study. Different approaches have been used to manage this challenge, including ignoring negative plate counts, using statistical estimations, or applying data transformations. Our objective was to illustrate and quantify the effect of negative plate count data management approaches on parameter estimation for microbial reduction models. Because it is impossible to obtain accurate plate counts below the LOD, we performed simulated experiments to generate synthetic data for both log-linear and Weibull-type microbial reductions. We then applied five different, previously reported data management practices and fit log-linear and Weibull models to the resulting data. The results indicated a significant effect (α = 0.05) of the data management practices on the estimated model parameters and performance indicators. For example, when the negative plate counts were replaced by the LOD for log-linear data sets, the slope of the subsequent log-linear model was, on average, 22% smaller than for the original data, the resulting model underpredicted lethality by up to 2.0 log, and the Weibull model was erroneously selected as the most likely correct model for those data. The results demonstrate that it is important to explicitly report LODs and related data management protocols, which can significantly affect model results, interpretation, and utility. Ultimately, we recommend using only the positive plate counts to estimate model parameters for microbial reduction curves and avoiding any data value substitutions or transformations when managing negative plate counts to yield the most accurate model parameters.

Microbial counts of mealworm larvae (Tenebrio molitor) and crickets (Acheta domesticus and Gryllodes sigillatus) from different rearing companies and different production batches.

PubMed

Vandeweyer, D; Crauwels, S; Lievens, B; Van Campenhout, L

2017-02-02

The rising interest in insects for human consumption and the changing regulations in Europe require a profound insight into the food safety of insects reared and sold in Western society. The microbial quality of edible insects has only been studied occasionally. This study aimed at generating an overview of intrinsic parameters (pH, water activity and moisture content) and microbial quality of fresh mealworm larvae and crickets for several rearing companies and for several batches per rearer. In total, 21 batches obtained from 7 rearing companies were subjected to analysis of intrinsic parameters, a range of plate counts and presence-absence tests for Salmonella spp. and Listeria monocytogenes. The microbial counts of the fresh insects were generally high. Different rearing batches from a single rearing company showed differences in microbial counts which could not be explained by variations in intrinsic properties. The largest variations were found in numbers of bacterial endospores, psychrotrophs and fungi. Salmonella spp. and L. monocytogenes were not detected in any of the samples. Altogether, our study shows that large variations were found between batches from individual rearers. As a consequence, no overall differences between rearers could be observed. Copyright © 2016 Elsevier B.V. All rights reserved.

Blood pressure cuffs as a vector for transmission of multi-resistant organisms: colonisation rates and effects of disinfection.

PubMed

Grewal, Harjeet; Varshney, Kavita; Thomas, Lee C; Kok, Jen; Shetty, Amith

2013-06-01

Blood pressure (BP) cuffs are potential vectors for transmission of multi-resistant organisms (MROs). The present study aims to determine MRO colonisation rates in BP cuffs from areas of high patient flow as an assessment of the quality of disinfection and infection control practices. BP cuffs in the ED, high dependency unit (HDU) and operating theatres (OT) were prospectively examined after routine disinfection procedures. Swabs collected from the inner and outer surfaces of BP cuffs during inter-patient intervals were plated onto replicate organism detection and counting, methicillin-resistant Staphylococcus aureus (MRSA) and vancomycin-resistant Enterococcus (VRE) chromogenic agar plates to detect rates of bacterial, MRSA and VRE colonisation, respectively. High bacterial colonisation rates were detected in BP cuffs from all three areas. BP cuffs from OT were significantly less colonised compared with cuffs from HDU and ED; 76% versus 96% and 100% (P < 0.0001) for inner surfaces and 86% versus 98% and 100% (P < 0.0001) for outer surfaces, respectively. Equivalent or higher bacterial growth was observed on the inner surface compared with outer surface in 54%, 84% and 86% of BP cuffs from OT, HDU and ED, respectively. MRSA was detected in 3 of 150 (2%) swabs collected, but no VRE was detected. Although MRSA and VRE were infrequently isolated, current disinfection and infection control protocols need to be improved given the greater recovery of organisms from the inner compared with outer surfaces of BP cuffs. © 2013 Australasian College for Emergency Medicine and Australasian Society for Emergency Medicine.

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.

Detection of Salmonella sp., Vibrio sp. and total plate count bacteria on blood cockle (Anadara granosa)

NASA Astrophysics Data System (ADS)

Ekawati, ER; Yusmiati, S. N. H.

2018-01-01

Blood cockle (Anadara granosa) has high level of zinc and protein, which is beneficial for therapeutic function for malnourished particularly stunting case in children. Zinc in animal foods is more absorbable than that from vegetable food. Blood cockle (Anadara granosa) is rich in nutrient and an excellent environment for the growth of microorganisms. This research aimed to identify the contamination of Salmonella sp., Vibrio sp. and total plate count bacteria on blood cockle (Anadara granosa). This was observation research with laboratory analysis. Salmonella sp. and Vibrio sp. were detected from blood cockle. Total plate count was determine of the total amount of the bacteria. Results detected from 20 samples of blood cockle showed that all samples were negative of Salmonella sp. and 1 sample positive Vibrio sp. The result of total plate count bacteria was < 5 x 105 colony/g sample.

Evaluation of free-stall mattress bedding treatments to reduce mastitis bacterial growth.

PubMed

Kristula, M A; Dou, Z; Toth, J D; Smith, B I; Harvey, N; Sabo, M

2008-05-01

Bacterial counts were compared in free-stall mattresses and teat ends exposed to 5 treatments in a factorial study design on 1 dairy farm. Mattresses in five 30-cow groups were subjected to 1 of 5 bedding treatments every other day: 0.5 kg of hydrated limestone, 120 mL of commercial acidic conditioner, 1 kg of coal fly ash, 1 kg of kiln-dried wood shavings, and control (no bedding). Counts of coliforms, Klebsiella spp., Escherichia coli, and Streptococcus spp. were lowest on mattresses bedded with lime. Mattresses bedded with the commercial acidic conditioner had the next lowest counts for coliforms, Klebsiella spp., and Streptococcus spp. Wood shavings and the no-bedding control had the highest counts for coliform and Klebsiella spp. Compared with wood shavings or control, fly ash reduced the counts of coliforms, whereas for the other 3 bacterial groups, the reduction was not always significant. Streptococcus spp. counts were greatest in the control group and did not differ among the shavings and fly ash groups. Teat swab results indicated that hydrated lime was the only bedding treatment that significantly decreased the counts of both coliforms and Klebsiella spp. There were no differences in Streptococcus spp. numbers on the teats between any of the bedding treatments. Bacterial populations grew steadily on mattresses and were generally higher at 36 to 48 h than at 12 to 24 h, whereas bacterial populations on teats grew rapidly by 12 h and then remained constant. Hydrated lime was the only treatment that significantly reduced bacterial counts on both mattresses and teat ends, but it caused some skin irritation.

Different binarization processes validated against manual counts of fluorescent bacterial cells.

PubMed

Tamminga, Gerrit G; Paulitsch-Fuchs, Astrid H; Jansen, Gijsbert J; Euverink, Gert-Jan W

2016-09-01

State of the art software methods (such as fixed value approaches or statistical approaches) to create a binary image of fluorescent bacterial cells are not as accurate and precise as they should be for counting bacteria and measuring their area. To overcome these bottlenecks, we introduce biological significance to obtain a binary image from a greyscale microscopic image. Using our biological significance approach we are able to automatically count about the same number of cells as an individual researcher would do by manual/visual counting. Using the fixed value or statistical approach to obtain a binary image leads to about 20% less cells in automatic counting. In our procedure we included the area measurements of the bacterial cells to determine the right parameters for background subtraction and threshold values. In an iterative process the threshold and background subtraction values were incremented until the number of particles smaller than a typical bacterial cell is less than the number of bacterial cells with a certain area. This research also shows that every image has a specific threshold with respect to the optical system, magnification and staining procedure as well as the exposure time. The biological significance approach shows that automatic counting can be performed with the same accuracy, precision and reproducibility as manual counting. The same approach can be used to count bacterial cells using different optical systems (Leica, Olympus and Navitar), magnification factors (200× and 400×), staining procedures (DNA (Propidium Iodide) and RNA (FISH)) and substrates (polycarbonate filter or glass). Copyright © 2016 Elsevier B.V. All rights reserved.

Bioorganic Fertilizer Enhances Soil Suppressive Capacity against Bacterial Wilt of Tomato

PubMed Central

Liu, Shuangri; Chai, Rushan; Huang, Weiqing; Liu, Xingxing; Tang, Caixian; Zhang, Yongsong

2015-01-01

Tomato bacterial wilt caused by Ralstonia solanacearum is one of the most destructive soil-borne diseases. Many strategies have been taken to improve soil suppressiveness against this destructive disease, but limited success has been achieved. In this study, a novel bioorganic fertilizer revealed a higher suppressive ability against bacterial wilt compared with several soil management methods in the field over four growing seasons from March 2011 to July 2013. The application of the bioorganic fertilizer significantly (P<0.05) reduced disease incidence of tomato and increased fruit yields in four independent trials. The association among the level of disease incidence, soil physicochemical and biological properties was investigated. The soil treated with the bioorganic fertilizer increased soil pH value, electric conductivity, organic carbon, NH4 +-N, NO3 --N and available K content, microbial activities and microbial biomass carbon content, which were positively related with soil suppressiveness. Bacterial and actinomycete populations assessed using classical plate counts were highest, whereas R. solanacearum and fungal populations were lowest in soil applied with the bioorganic fertilizer. Microbial community diversity and richness were assessed using denaturing gel gradient electrophoresis profile analysis. The soil treated with the bioorganic fertilizer exhibited higher bacterial community diversity but lower fungal community diversity. Redundancy analysis showed that bacterial community diversity and richness negatively related with bacterial wilt suppressiveness, while fungal community richness positively correlated with R. solanacearum population. We concluded that the alteration of soil physicochemical and biological properties in soil treated with the bioorganic fertilizer induced the soil suppressiveness against tomato bacterial wilt. PMID:25830639

New evidence for Cu-decorated binary-oxides mediating bacterial inactivation/mineralization in aerobic media.

PubMed

Rtimi, S; Pulgarin, C; Bensimon, M; Kiwi, J

2016-08-01

Binary oxide semiconductors TiO2-ZrO2 and Cu-decorated TiO2-ZrO2 (TiO2-ZrO2-Cu) uniform films were sputtered on polyester (PES). These films were irradiated under low intensity solar simulated light and led to bacterial inactivation in aerobic and anaerobic media as evaluated by CFU-plate counting. But bacterial mineralization was only induced by TiO2-ZrO2-Cu in aerobic media. The highly oxidative radicals generated on the films surface under light were identified by the use of appropriate scavengers. The hole generated on the TiO2-ZrO2 films is shown to be the main specie leading to bacterial inactivation. TiO2-ZrO2 and Cu-decorated TiO2-ZrO2 films release Zr and Ti <1ppb and Cu 4.6ppb/cm(2) as determined by inductively coupled plasma mass spectrometry (ICP-MS) This level is far below the citotoxicity permitted level allowed for mammalian cells suggesting that bacterial disinfection proceeds through an oligodynamic effect. By Fourier transform attenuated infrared spectroscopy (ATR-FTIR) the systematic shift of the predominating νs(CH2) vibrational-rotational peak making up most of the bacterial cell-wall content in C was monitored. Based on this evidence a mechanism suggested leading to CH bond stretching followed by cell lysis and cell death. Bacterial inactivation cycling was observed on TiO2-ZrO2-Cu showing the stability of these films leading to bacterial inactivation. Copyright © 2016 Elsevier B.V. All rights reserved.

Death of the Escherichia coli K-12 strain W3110 in soil and water.

PubMed Central

Bogosian, G; Sammons, L E; Morris, P J; O'Neil, J P; Heitkamp, M A; Weber, D B

1996-01-01

Whether Escherichia coli K-12 strain W3110 can enter the "viable but nonculturable" state was studied with sterile and nonsterile water and soil at various temperatures. In nonsterile river water, the plate counts of added E. coli cells dropped to less than 10 CFU/ml in less than 10 days. Acridine orange direct counts, direct viable counts, most-probable-number estimates, and PCR analyses indicated that the added E. coli cells were disappearing from the water in parallel with the number of CFU. Similar results were obtained with nonsterile soil, although the decline of the added E. coli was slower. In sterile water or soil, the added E. coli persisted for much longer, often without any decline in the plate counts even after 50 days. In sterile river water at 37 degrees C and sterile artificial seawater at 20 and 37 degrees C, the plate counts declined by 3 to 5 orders of magnitude, while the acridine orange direct counts remained unchanged. However, direct viable counts and various resuscitation studies all indicated that the nonculturable cells were nonviable. Thus, in either sterile or nonsterile water and soil, the decline in plate counts of E. coli K-12 strain W3110 is not due to the cells entering the viable but nonculturable state, but is simply due to their death. PMID:8900002

Effectiveness of Polyvalent Bacterial Lysate and Autovaccines Against Upper Respiratory Tract Bacterial Colonization by Potential Pathogens: A Randomized Study

PubMed Central

Zagólski, Olaf; Stręk, Paweł; Kasprowicz, Andrzej; Białecka, Anna

2015-01-01

Background Polyvalent bacterial lysate (PBL) is an oral immunostimulating vaccine consisting of bacterial standardized lysates obtained by lysis of different strains of bacteria. Autovaccines are individually prepared based on the results of smears obtained from the patient. Both types of vaccine can be used to treat an ongoing chronic infection. This study sought to determine which method is more effective against nasal colonization by potential respiratory tract pathogens. Material/Methods We enrolled 150 patients with aerobic Gram stain culture and count results indicating bacterial colonization of the nose and/or throat by potential pathogens. The participants were randomly assigned to each of the following groups: 1. administration of PBL, 2. administration of autovaccine, and 3. no intervention (controls). Results Reduction of the bacterial count in Streptococcus pneumoniae-colonized participants was significant after the autovaccine (p<0.001) and PBL (p<0.01). Reduction of the bacterial count of other β-hemolytic streptococcal strains after treatment with the autovaccine was significant (p<0.01) and was non-significant after PBL. In Haemophilus influenzae colonization, significant reduction in the bacterial count was noted in the PBL group (p<0.01). Methicillin-resistant Staphylococcus aureus colonization did not respond to either treatment. Conclusions The autovaccine is more effective than PBL for reducing bacterial count of Streptococcus pneumoniae and β-hemolytic streptococci, while PBL was more effective against Haemophilus influenzae colonization. PMID:26434686

A selective medium for the enumeration and differentiation of Lactobacillus delbrueckii ssp. bulgaricus.

PubMed

Nwamaioha, Nwadiuto O; Ibrahim, Salam A

2018-06-01

Modified reinforced clostridial medium (mRCM) was developed and evaluated for the differential enumeration of Lactobacillus delbrueckii ssp. bulgaricus. Lactobacillus bulgaricus, an important species of lactic acid bacteria with health benefits, is used in the production of yogurt and other fermented foods. Our results showed that supplementing reinforced clostridial medium with 0.025% CaCl 2 , 0.01% uracil, and 0.2% Tween 80 (mRCM) significantly enhanced the growth rate of L. bulgaricus RR and ATCC 11842 strains as measured by the optical densities of these strains after 12 h of incubation at 42°C. The bacterial populations (plate count) of the RR and ATCC 11842 strains were 0.76 and 0.77 log cfu/g higher in mRCM than in de Man, Rogosa, and Sharpe and reinforced clostridial medium media, respectively. Conversely, the population counts for other bacterial species (Bifidobacterium, Lactobacillus rhamnosus, and Lactobacillus reuteri) were significantly inhibited in the mRCM medium. The addition of aniline blue dye to mRCM (mRCM-blue) improved the selectivity of L. bulgaricus in mixed lactic bacterial cultures compared with de Man, Rogosa, and Sharpe medium and lactic agar with regard to colony appearance and morphology. The mRCM-blue performed better than the conventional medium in culturing, enumerating, and differentiating L. bulgaricus. Therefore, mRCM-blue could be used as a selective medium to enhance the growth and differentiation of L. bulgaricus in order to meet the increasing demand for this beneficial species of bacteria. Copyright © 2018 American Dairy Science Association. Published by Elsevier Inc. All rights reserved.

The diabetes nutrition education study randomized controlled trial: A comparative effectiveness study of approaches to nutrition in diabetes self-management education.

PubMed

Bowen, Michael E; Cavanaugh, Kerri L; Wolff, Kathleen; Davis, Dianne; Gregory, Rebecca P; Shintani, Ayumi; Eden, Svetlana; Wallston, Ken; Elasy, Tom; Rothman, Russell L

2016-08-01

To compare the effectiveness of different approaches to nutrition education in diabetes self-management education and support (DSME/S). We randomized 150 adults with type 2 diabetes to either certified diabetes educator (CDE)-delivered DSME/S with carbohydrate gram counting or the modified plate method versus general health education. The primary outcome was change in HbA1C over 6 months. At 6 months, HbA1C improved within the plate method [-0.83% (-1.29, -0.33), P<0.001] and carbohydrate counting [-0.63% (-1.03, -0.18), P=0.04] groups but not the control group [P=0.34]. Change in HbA1C from baseline between the control and intervention groups was not significant at 6 months (carbohydrate counting, P=0.36; modified plate method, P=0.08). In a pre-specified subgroup analysis of patients with a baseline HbA1C 7-10%, change in HbA1C from baseline improved in the carbohydrate counting [-0.86% (-1.47, -0.26), P=0.006] and plate method groups [-0.76% (-1.33, -0.19), P=0.01] compared to controls. CDE-delivered DSME/S focused on carbohydrate counting or the modified plate method improved glycemic control in patients with an initial HbA1C between 7 and 10%. Both carbohydrate counting and the modified plate method improve glycemic control as part of DSME/S. Copyright © 2016 Elsevier Ireland Ltd. All rights reserved.

Aseptic Laboratory Techniques: Plating Methods

PubMed Central

Sanders, Erin R.

2012-01-01

Microorganisms are present on all inanimate surfaces creating ubiquitous sources of possible contamination in the laboratory. Experimental success relies on the ability of a scientist to sterilize work surfaces and equipment as well as prevent contact of sterile instruments and solutions with non-sterile surfaces. Here we present the steps for several plating methods routinely used in the laboratory to isolate, propagate, or enumerate microorganisms such as bacteria and phage. All five methods incorporate aseptic technique, or procedures that maintain the sterility of experimental materials. Procedures described include (1) streak-plating bacterial cultures to isolate single colonies, (2) pour-plating and (3) spread-plating to enumerate viable bacterial colonies, (4) soft agar overlays to isolate phage and enumerate plaques, and (5) replica-plating to transfer cells from one plate to another in an identical spatial pattern. These procedures can be performed at the laboratory bench, provided they involve non-pathogenic strains of microorganisms (Biosafety Level 1, BSL-1). If working with BSL-2 organisms, then these manipulations must take place in a biosafety cabinet. Consult the most current edition of the Biosafety in Microbiological and Biomedical Laboratories (BMBL) as well as Material Safety Data Sheets (MSDS) for Infectious Substances to determine the biohazard classification as well as the safety precautions and containment facilities required for the microorganism in question. Bacterial strains and phage stocks can be obtained from research investigators, companies, and collections maintained by particular organizations such as the American Type Culture Collection (ATCC). It is recommended that non-pathogenic strains be used when learning the various plating methods. By following the procedures described in this protocol, students should be able to: ● Perform plating procedures without contaminating media. ● Isolate single bacterial colonies by the streak-plating method. ● Use pour-plating and spread-plating methods to determine the concentration of bacteria. ● Perform soft agar overlays when working with phage. ● Transfer bacterial cells from one plate to another using the replica-plating procedure. ● Given an experimental task, select the appropriate plating method. PMID:22617405

Evaluation of the sealing capability of implants to titanium and zirconia abutments against Porphyromonas gingivalis, Prevotella intermedia, and Fusobacterium nucleatum under different screw torque values.

PubMed

Smith, Nicole A; Turkyilmaz, Ilser

2014-09-01

When evaluating long-term implant success, clinicians have always been concerned with the gap at the implant-abutment junction, where bacteria can accumulate and cause marginal bone loss. However, little information regarding bacterial leakage at the implant-abutment junction, or microgap, is available. The purpose of this study was to evaluate sealing at 2 different implant-abutment interfaces under different screw torque values. Twenty sterile zirconia abutments and 20 sterile titanium abutments were screwed into 40 sterile implants and placed in test tubes. The ability of a bacterial mixture of Prevotella intermedia, Porphyromonas gingivalis, and Fusobacterium nucleatum to leak through an implant-titanium abutment seal under 20 and 35 Ncm torque values and an implant-zirconia abutment seal under 20 and 35 Ncm torque values was evaluated daily until leakage was noted. Once a unit demonstrated leakage, a specimen was plated. After 4 days, the number of colonies on each plate was counted with an electronic colony counter. Plating was used to verify whether or not bacterial leakage occurred and when leakage first occurred. The implant-abutment units were removed and rinsed with phosphate buffered saline solution and evaluated with a stereomicroscope. The marginal gap between the implant and the abutment was measured and correlated with the amount of bacterial leakage. The data were analyzed with ANOVA. Bacterial leakage was noted in all specimens, regardless of material or screw torque value. With titanium abutments, changing the screw torque value from 20 to 35 Ncm did not significantly affect the amount of bacterial leakage. However, with zirconia abutments, changing the screw torque value from 20 to 35 Ncm was statistically significant (P<.017). Overall, the marginal gap noted was larger at the zirconia-abutment interface (5.25 ±1.99 μm) than the titanium-abutment interface (12.38 ±3.73 μm), irrespective of the screw torque value. Stereomicroscopy revealed a nonuniform marginal gap in all specimens. The results of this study showed that, over time, bacteria will leak through the implant-abutment microgap at the implant-abutment interface. Implants with a titanium abutment demonstrate a smaller microgap than implants with a zirconia abutment. Tightening the zirconia abutment screw from 20 to 35 Ncm decreases the size of the microgap, which suggests a more intimate fit between the implant and the abutment. Copyright © 2014 Editorial Council for the Journal of Prosthetic Dentistry. Published by Elsevier Inc. All rights reserved.

Effect of face washing with soap and water and cleaning with antiseptics on upper-lid bacteria of surgical eye patients.

PubMed

Bekibele, Charles O; Kehinde, Aderemi O; Ajayi, Benedictus G K

2010-12-01

To determine the effect of face washing with soap and water and cleaning with povidone iodine and cetrimide/chlorhexidine gluconate (Savlon) on upper-lid bacteria. Prospective, nonrandomized clinical trial. Eighty patients attending the Eye Clinic, University College Hospital, Ibadan, Nigeria. Eighty patients assigned to 4 groups had swabs of the upper eyelid skin taken before and after face wash with soap and water, and cleansing with Savlon and 5% povidone iodine. Specimens were cultured and Gram stained. Bacterial counts were carried out using standard techniques. Face washing with soap and water increased the proportion of patients with bacterial isolates from 80.0% to 87.5%. The average colony count increased from 187.1 to 318.5 colony units per mL (p = 0.02). Application of 5% povidone iodine without face washing with soap and water reduced the proportion of patients with bacterial isolates from 82.6% (mean count 196.5) to 28.6% (mean count 34.1)(p = 0.001); in comparison, the application of 5% povidone iodine after face washing with soap and water reduced the proportion from 71.4% (mean count 133.9) to 40.0% (mean count 69.0)(p = 0.01). Application of Savlon without face washing with soap and water reduced the proportion of patients with bacterial isolates from 100% (mean count 310.9) to 41.2% (mean count 19.8)(p = 0.004) compared with the application after face washing, which reduced the proportion from 89.5% (mean count 240.3) to 41.2% (mean count 82.9)(p = 0.02). Both povidone and Savlon are effective in reducing periocular bacteria in an African setting. Prior face washing with soap and water had no added benefit in reducing bacterial colony count.

Zinc-ion implanted and deposited titanium surfaces reduce adhesion of Streptococccus mutans

NASA Astrophysics Data System (ADS)

Xu, Juan; Ding, Gang; Li, Jinlu; Yang, Shenhui; Fang, Bisong; Sun, Hongchen; Zhou, Yanmin

2010-10-01

While titanium (Ti) is a commonly used dental implant material with advantageous biocompatible and mechanical properties, native Ti surfaces do not have the ability to prevent bacterial colonization. The objective of this study was to evaluate the chemical composition and bacterial adhesive properties of zinc (Zn) ion implanted and deposited Ti surfaces (Zn-PIIID-Ti) as potential dental implant materials. Surfaces of pure Ti (cp-Ti) were modified with increasing concentrations of Zn using plasma immersion ion implantation and deposition (PIIID), and elemental surface compositions were characterized by X-ray photoelectron spectrometry (XPS). To evaluate bacterial responses, Streptococcus mutans were seeded onto the modifiedTi surfaces for 48 h and subsequently observed by scanning electron microscopy. Relative numbers of bacteria on each surface were assessed by collecting the adhered bacteria, reculturing and counting colony forming units after 48 h on bacterial grade plates. Ti, oxygen and carbon elements were detected on all surfaces by XPS. Increased Zn signals were detected on Zn-PIIID-Ti surfaces, correlating with an increase of Zn-deposition time. Substantial numbers of S. mutans adhered to cp-Ti samples, whereas bacterial adhesion on Zn-PIIID-Ti surfaces signficantly decreased as the Zn concentration increased ( p < 0.01). In conclusion, PIIID can successfully introduce Zn onto a Ti surface, forming a modified surface layer bearing Zn ions that consequently deter adhesion of S. mutans, a common bacterium in the oral environment.

Prevention and control of carbapenemase-producing organisms at a regional burns centre.

PubMed

Teare, L; Myers, J; Kirkham, A; Tredoux, T; Martin, R; Boasman, S; Wisbey, A; Charlton, C; Dziewulski, P

2016-06-01

In many parts of the world, carbapenemase-producing organisms (CPOs) are endemic. The transfer of medical patients from such countries to the UK requires us to have control systems in place to avoid onward transmission. This report describes the experience of a regional burns centre challenged by its first four cases of CPO in two separate incidents. Key learning from our experience was the importance of CPOs being considered in empirical antibiotics for any patient from an endemic area. Using contact plates, we demonstrated high bacterial counts after cleaning and we describe a terminal cleaning strategy along with the importance of continuing staff engagement and education. Copyright © 2016. Published by Elsevier Ltd.

Evaluation of procalcitonin and neopterin level in serum of patients with acute bacterial infection.

PubMed

Pourakbari, Babak; Mamishi, Setareh; Zafari, Javid; Khairkhah, Hanieh; Ashtiani, Mohammad H; Abedini, Masomeh; Afsharpaiman, Shahla; Rad, Soroush Seifi

2010-01-01

Fever as a common presenting complaint in pediatric patients can be due to various causes. Differentiating bacterial infection from other causes is important because the prompt use of antibiotics is critical in bacterial infection. Traditional markers of infection such as BT and WBC count may be unspecific and culture may be late or absent. CRP and Procalcitonin (PCT) have been considered to evaluate the evolution of infections and sepsis in patients presenting with SIRS. Neopterin has also been proposed to aid in the diagnosis of bacterial infection. In this study, we compared the value of the serum PCT, neopterin level, and WBC count for predicting bacterial infection and outcome in children with fever. 158 pediatric (2-120-month-old) patients suspected to have acute bacterial infection, based on clinical judgment in which other causes of SIRS were ruled out were included in the study. WBC count with differential was determined and PCT and neopterin levels were measured. PCT level was higher in bacterial infection and patients who were complicated or expired. Rapid PCT test is superior to neopterin and WBC count for anticipating bacterial infection, especially in ED where prompt decision making is critical.

Adherence of Aeromonas hydrophila strains to human enterocyte-like cells pre-infected with rotavirus.

PubMed

Bertuccio, M P; Picerno, I; Scoglio, M E

2012-09-01

The interest grown in these years about emerging pathogens in the onset of intestinal disease showed that the pathogenic mechanism is a multifactorial event. Our objective was to evaluate the role of co-infection with rotavirus in the expression of Aeromonas spp adhesiveness. The rate of co-infection involves contact of Caco-2 cells with the virus, followed by adsorption for 1 and 2 hours. Aliquots of bacterial suspensions were added to tissue-culture plates. After infection, cell monolayers were lysed; serially diluted lysates were plated to determine the number of bound bacteria by performing colony forming units (CFU) counts. Non-adhesive strains were not subject to variations resulting from co-infection, while those who had medium or high adhesiveness gave rise to an increase of the same. Infection with rotavirus promotes the Aeromonas ability to adhere to Caco-2 cells and this effect depends on the duration of infection and on the starting adhesiveness of bacteria strain.

Fluoro-luminometric real-time measurement of bacterial viability and killing.

PubMed

Lehtinen, Janne; Virta, Marko; Lilius, Esa Matti

2003-10-01

The viability and killing of Escherichia coli was measured on a real-time basis using a fluoro-luminometric device, which allows successive measurements of fluorescence and bioluminescence without user intervention. Bacteria were made fluorescent and bioluminescent by expression of gfp and insect luciferase (lucFF) genes. The green fluorescent protein (GFP) is a highly fluorescent, extremely stable protein, which accumulates in cells during growth, and therefore the measured fluorescence signal was proportional to the total number of cells. The luciferase reaction is dependent of ATP produced by living cells, so that the bioluminescence level was a direct measure of the viable cells. In contrast to the bacterial luciferase, the insect luciferase uses a water-soluble and nonvolatile substrate, which makes automated multi-well microplate assay possible. For the validation of the assay, the proportion of living and dead cell populations was experimentally modified by incubating E. coli cells in the presence of various ethanol concentrations. Bacterial viability and killing measured by a fluoro-luminometric assay correlated fairly well with the reference methods: conventional plate counting, optical density measurement and various flow cytometric analyses. The real-time assay described here allows following the changes in bacterial cultures and assessing the bactericidal and other effects of various chemical, immunological and physical agents simultaneously in large numbers of samples.

Bacterial communities in the collection and chlorinated distribution sections of a drinking water system in Budapest, Hungary.

PubMed

Homonnay, Zalán G; Török, György; Makk, Judit; Brumbauer, Anikó; Major, Eva; Márialigeti, Károly; Tóth, Erika

2014-07-01

Bacterial communities of a bank-filtered drinking water system were investigated by aerobic cultivation and clone library analysis. Moreover, bacterial communities were compared using sequence-aided terminal restriction fragment length polymorphism (T-RFLP) fingerprinting at ten characteristic points located at both the collecting and the distributing part of the water supply system. Chemical characteristics of the samples were similar, except for the presence of chlorine residuals in the distribution system and increased total iron concentration in two of the samples. Assimilable organic carbon (AOC) concentration increased within the collection system, it was reduced by chlorination and it increased again in the distribution system. Neither fecal indicators nor pathogens were detected by standard cultivation techniques. Chlorination reduced bacterial diversity and heterotrophic plate counts. Community structures were found to be significantly different before and after chlorination: the diverse communities in wells and the collection system were dominated by chemolithotrophic (e.g., Gallionella and Nitrospira) and oligocarbophilic-heterotrophic bacteria (e.g., Sphingomonas, Sphingopyxis, and Bradyrhizobium). After chlorination in the distribution system, the most characteristic bacterium was related to the facultative methylotrophic Methylocella spp. Communities changed within the distribution system too, Mycobacterium spp. or Sphingopyxis spp. became predominant in certain samples. © 2014 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

Real-time bacterial microcolony counting using on-chip microscopy

NASA Astrophysics Data System (ADS)

Jung, Jae Hee; Lee, Jung Eun

2016-02-01

Observing microbial colonies is the standard method for determining the microbe titer and investigating the behaviors of microbes. Here, we report an automated, real-time bacterial microcolony-counting system implemented on a wide field-of-view (FOV), on-chip microscopy platform, termed ePetri. Using sub-pixel sweeping microscopy (SPSM) with a super-resolution algorithm, this system offers the ability to dynamically track individual bacterial microcolonies over a wide FOV of 5.7 mm × 4.3 mm without requiring a moving stage or lens. As a demonstration, we obtained high-resolution time-series images of S. epidermidis at 20-min intervals. We implemented an image-processing algorithm to analyze the spatiotemporal distribution of microcolonies, the development of which could be observed from a single bacterial cell. Test bacterial colonies with a minimum diameter of 20 μm could be enumerated within 6 h. We showed that our approach not only provides results that are comparable to conventional colony-counting assays but also can be used to monitor the dynamics of colony formation and growth. This microcolony-counting system using on-chip microscopy represents a new platform that substantially reduces the detection time for bacterial colony counting. It uses chip-scale image acquisition and is a simple and compact solution for the automation of colony-counting assays and microbe behavior analysis with applications in antibacterial drug discovery.

Real-time bacterial microcolony counting using on-chip microscopy

PubMed Central

Jung, Jae Hee; Lee, Jung Eun

2016-01-01

Observing microbial colonies is the standard method for determining the microbe titer and investigating the behaviors of microbes. Here, we report an automated, real-time bacterial microcolony-counting system implemented on a wide field-of-view (FOV), on-chip microscopy platform, termed ePetri. Using sub-pixel sweeping microscopy (SPSM) with a super-resolution algorithm, this system offers the ability to dynamically track individual bacterial microcolonies over a wide FOV of 5.7 mm × 4.3 mm without requiring a moving stage or lens. As a demonstration, we obtained high-resolution time-series images of S. epidermidis at 20-min intervals. We implemented an image-processing algorithm to analyze the spatiotemporal distribution of microcolonies, the development of which could be observed from a single bacterial cell. Test bacterial colonies with a minimum diameter of 20 μm could be enumerated within 6 h. We showed that our approach not only provides results that are comparable to conventional colony-counting assays but also can be used to monitor the dynamics of colony formation and growth. This microcolony-counting system using on-chip microscopy represents a new platform that substantially reduces the detection time for bacterial colony counting. It uses chip-scale image acquisition and is a simple and compact solution for the automation of colony-counting assays and microbe behavior analysis with applications in antibacterial drug discovery. PMID:26902822

An anti-bacterial approach to nanoscale roughening of biomimetic rice-like pattern PP by thermal annealing

NASA Astrophysics Data System (ADS)

Jafari Nodoushan, Emad; Ebrahimi, Nadereh Golshan; Ayazi, Masoumeh

2017-11-01

In this paper, we introduced thermal annealing treatment as an effective way of increasing the nanoscale roughness of a semi-crystalline polymer surface. Annealing treatment applied to a biomimetic microscale pattern of rice leaf to achieve a superhydrophobic surface with a hierarchical roughness. Resulted surfaces was characterized by XRD, AFM and FE-SEM instruments and showed an increase of roughness and cristallinity within both time and temperature of treatment. These two parameters also impact on measured static contact angle up to 158°. Bacterial attachment potency has an inverse relationship with the similarity of surface pattern dimensions and bacterial size and due to that, thermal annealing could be an effective way to create anti-bacterial surface beyond its effect on water repellency. Point in case, the anti-bacterial properties of produced water-repellence surfaces of PP were measured and counted colonies of both gram-negative (E. coli) and gram-positive (S. aureus) bacteria reduced with the nature of PP and hierarchical pattern on that. Anti-bacterial characterization of the resulted surface reveals a stunning reduction in adhesion of gram-positive bacteria to the surface. S. aureus reduction rates equaled to 95% and 66% when compared to control blank plate and smooth surface of PP. Moreover, it also could affect the other type of bacteria, gram-negative (E. coli). In the latter case, adhesion reduction rates calculated 66% and 53% when against to the same controls, respectively.

[Study of physicochemical and bacteriological impact of a weekly assembly of automated compounding BAXA(®) Exacta-Mix 2400 on parenteral nutrition admixture manufacturing].

PubMed

Chappuy, L; Charroin, C; Vételé, F; Durand, T; Quessada, T; Klotz, M-C; Bréant, V; Aulagner, G

2014-01-01

The parenteral nutrition admixtures are manufactured with an automated compounding BAXA(®) Exacta-Mix 2400. A 48-hour assembly has been validated. To optimize time and cost, a weekly assembly was tested. Assembly was made on the first day. Ten identical parenteral nutrition admixtures (different volumes and compositions) were produced each day. A macroscopic examination was done at D0, D7 and D14. Physicochemical controls (electrolytes determinations by atomic absorption spectrophotometry, osmolalities measurements) were performed. Microbiological tests included a filtration membrane sterility test (Steritest(®)) and a plate count agar environmental monitoring. All mixtures were considered stable. The 12 Steritest(®) (H24, H48, D7 and D14) did not show any bacterial or fungal contamination. No microorganism has been detected on the plate count agar at D4 and D7. Concerning the physicochemical parameters of each parental nutrition admixture, no significant difference (Wilcoxon test) with the first day was found. The automated filling system BAXA(®) Exacta-Mix 2400 improves the quality and safety of production. According to these results, the weekly assembly is validated and permit to save time (80hours/year) and cost (40 000 euros on consumable/year). Copyright © 2013 Elsevier Masson SAS. All rights reserved.

Evaluation of free-stall mattress bedding treatments to reduce mastitis bacterial growth

DOE Office of Scientific and Technical Information (OSTI.GOV)

Kristula, M.A.; Dou, Z.; Toth, J.D.

2008-05-15

Bacterial counts were compared in free-stall mattresses and teat ends exposed to 5 treatments in a factorial study design on 1 dairy farm. Mattresses in five 30-cow groups were subjected to 1 of 5 bedding treatments every other day: 0.5 kg of hydrated limestone, 120 mL of commercial acidic conditioner, 1 kg of coal fly ash, 1 kg of kiln-dried wood shavings, and control (no bedding). Counts of coliforms, Klebsiella spp., Escherichia coli, and Streptococcus spp. were lowest on mattresses bedded with lime. Mattresses bedded with the commercial acidic conditioner had the next lowest counts for coliforms, Klebsiella spp., andmore » Streptococcus spp. Wood shavings and the no-bedding control had the highest counts for coliform and Klebsiella spp. Compared with wood shavings or control, fly ash reduced the counts of coliforms, whereas for the other 3 bacterial groups, the reduction was not always significant. Streptococcus spp. counts were greatest in the control group and did not differ among the shavings and fly ash groups. Teat swab results indicated that hydrated lime was the only bedding treatment that significantly decreased the counts of both coliforms and Klebsiella spp. There were no differences in Streptococcus spp. numbers on the teats between any of the bedding treatments. Bacterial populations grew steadily on mattresses and were generally higher at 36 to 48 h than at 12 to 24 h, whereas bacterial populations on teats grew rapidly by 12 h and then remained constant. Hydrated lime was the only treatment that significantly reduced bacterial counts on both mattresses and teat ends, but it caused some skin irritation.« less

Bifidobacterium breve IPLA20005 affects in vitro the expression of hly and luxS genes, related to the virulence of Listeria monocytogenes Lm23.

PubMed

Rios-Covian, David; Nogacka, Alicja; Salazar, Nuria; Hernández-Barranco, A M; Cuesta, Isabel; Gueimonde, Miguel; de Los Reyes Gavilán, Clara G

2018-03-01

Mechanistic features that characterize the interaction and inhibition of the food-borne pathogen Listeria monocytogenes by members of the genus Bifidobacterium still remain unclear. In the present work, we tried to shed light on the influence that co-cultivation of L. monocytogenes with Bifidobacterium breve may exert on both microorganisms and on virulence of the pathogen. Production of acetate and lactate was measured by gas chromatography and high-performance liquid chromatography, respectively; bacterial counts were obtained by plate count; gene expression was determined by RT-qPCR; and haemolytic activity was analyzed against goat erythrocytes. We found slightly but significantly lower final counts of Listeria and Bifidobacterium (p < 0.05) and lower haemolytic efficiency in L. monocytogenes cells from cocultures than in those from monocultures. In contrast, the hly and luxS genes, which code for the cytolysin listeriolysin O and participate in biofilm formation, respectively, were overexpressed when L. monocytogenes was grown in coculture. This indicates that the presence of Bifidobacterium is able to modify the gene expression and haemolytic activity of L. monocytogenes when both microorganisms grow together.

Bacterial contaminants in carbonated soft drinks sold in Bangladesh markets.

PubMed

Akond, Muhammad Ali; Alam, Saidul; Hasan, S M R; Mubassara, Sanzida; Uddin, Sarder Nasir; Shirin, Momena

2009-03-31

A total of 225 carbonated soft drink (CSD) samples from nine brands, from various locations in five metropolitan cities of Bangladesh were examined to determine their bacteriological quality. Most samples were not in compliance with microbiological standards set by organizations like the World Health Organization (WHO). Pseudomonas aeruginosa was the predominant species with an incidence of 95%. Streptococcus spp. and Bacillus stearothermophilus were the next most prevalent with numbers ranging from 6 to 122 and 9 to 105 cfu/100 ml, respectively. Fifty four percent of the samples yielded Salmonella spp. at numbers ranging from 2 to 90 cfu/100 ml. Total coliform (TC) and faecal coliform (FC) counts were found in 68-100% and 76-100% of samples of individual brands, at numbers ranging from 5 to 213 and 3 to 276 cfu/100 ml, respectively. According to WHO standards 60-88% of samples from six brands and 32% and 40% of samples from two other brands belonged to the intermediate risk group with FC counts of 100-1000 cfu/100 ml. Heterotrophic plate counts, however, were under the permissible limit in all 225 samples. These findings suggest that carbonated soft drinks commercially available in Bangladesh pose substantial risks to public health.

Modulation of cell surface hydrophobicity and attachment of bacteria to abiotic surfaces and shrimp by Malaysian herb extracts.

PubMed

Hui, Yew Woh; Dykes, Gary A

2012-08-01

The use of simple crude water extracts of common herbs to reduce bacterial attachment may be a cost-effective way to control bacterial foodborne pathogens, particularly in developing countries. The ability of water extracts of three common Malaysian herbs (Andrographis paniculata, Eurycoma longifolia, and Garcinia atroviridis) to modulate hydrophobicity and attachment to surfaces of five food-related bacterial strains (Bacillus cereus ATCC 14576, Escherichia coli ATCC 25922, Pseudomonas aeruginosa ATCC 10145, Salmonella Enteritidis ATCC 13076, Staphylococcus aureus ATCC 25923) were determined. The bacterial attachment to hydrocarbon assay was used to determine bacterial hydrophobicity. Staining and direct microscopic counts were used to determine attachment of bacteria to glass and stainless steel. Plating on selective media was used to determine attachment of bacteria to shrimp. All extracts were capable of either significantly ( P < 0.05) increasing or decreasing bacterial surface hydrophobicity, depending on the herb extract and bacteria combination. Bacterial attachment to all surfaces was either significantly (P < 0.05) increased or decreased, depending on the herb extract and bacteria combination. Overall, hydrophobicity did not show a significant correlation (P > 0.05) to bacterial attachment. For specific combinations of bacteria, surface material, and plant extract, significant correlations (R > 0.80) between hydrophobicity and attachment were observed. The highest of these was observed for S. aureus attachment to stainless steel and glass after treatment with the E. longifolia extract (R = 0.99, P < 0.01). The crude water herb extracts in this study were shown to have the potential to modulate specific bacterial and surface interactions and may, with further work, be useful for the simple and practical control of foodborne pathogens.

AUTOMATIC COUNTER

DOEpatents

Robinson, H.P.

1960-06-01

An automatic counter of alpha particle tracks recorded by a sensitive emulsion of a photographic plate is described. The counter includes a source of mcdulated dark-field illumination for developing light flashes from the recorded particle tracks as the photographic plate is automatically scanned in narrow strips. Photoelectric means convert the light flashes to proportional current pulses for application to an electronic counting circuit. Photoelectric means are further provided for developing a phase reference signal from the photographic plate in such a manner that signals arising from particle tracks not parallel to the edge of the plate are out of phase with the reference signal. The counting circuit includes provision for rejecting the out-of-phase signals resulting from unoriented tracks as well as signals resulting from spurious marks on the plate such as scratches, dust or grain clumpings, etc. The output of the circuit is hence indicative only of the tracks that would be counted by a human operator.

Influence of preservation methods on the quality of colostrum sourced from New Zealand dairy farms.

PubMed

Denholm, K S; Hunnam, J C; Cuttance, E L; McDougall, S

2017-09-01

To assess the effect of two temperatures (ambient temperature and 4°C), three preservation methods (no preservative, yoghurt and potassium sorbate), and two periods of storage (3 and 7 days) on Brix and total bacterial and coliform counts of colostrum collected from New Zealand dairy farms. One litre of colostrum destined to be fed to newborn calves was collected from 55 New Zealand dairy farms in the spring of 2015. Six aliquots of 150 mL were obtained from each colostrum sample, with two aliquots left untreated, two treated with potassium sorbate and two with yoghurt, and one of each pair of aliquots stored at ambient temperature and the other at 4°C. All samples were tested for Brix, total bacterial counts and coliform counts before treatment (Day 0), and after 3 and 7 days of storage. The effect of preservation method and storage temperature on the change in Brix, bacterial and coliform counts after 3 or 7 days of storage was analysed using multivariable random effects models. For all outcome variables there was a temperature by preservation interaction. For aliquots preserved with potassium sorbate, changes in Brix and bacterial counts did not differ between aliquots stored at ambient temperature or 4°C, but for aliquots preserved with yoghurt or no preservative the decrease in Brix and increase in bacterial counts was greater for aliquots stored at ambient temperature than 4°C (p<0.001). For aliquots preserved with potassium sorbate, coliform counts decreased at both temperatures, but for aliquots preserved with yoghurt or no preservative coliform counts increased for aliquots stored at 4°C, but generally decreased at ambient temperatures (p<0.001). There was also an interaction between duration of storage and temperature for bacterial counts (p<0.001). The difference in the increase in bacterial counts between aliquots stored at 4°C and ambient temperature after 3 days was greater than between aliquots stored at 4°C and ambient temperature after 7 days. Use of potassium sorbate to preserve colostrum for 3 or 7 days resulted in little or no reduction in Brix and a lower increase in total bacterial counts than colostrum stored without preservative or with yoghurt added. Colostrum quality was not affected by storage temperature for samples preserved with potassium sorbate, but storage at 4°C resulted in better quality colostrum than storage at ambient temperatures for colostrum with no preservative or yoghurt added.

Mechanical reduction of the intracanal Enterococcus faecalis population by Hyflex CM, K3XF, ProTaper Next, and two manual instrument systems: an in vitro comparative study.

PubMed

Tewari, Rajendra K; Ali, Sajid; Mishra, Surendra K; Kumar, Ashok; Andrabi, Syed Mukhtar-Un-Nisar; Zoya, Asma; Alam, Sharique

2016-05-01

In the present study, the effectiveness of three rotary and two manual nickel titanium instrument systems on mechanical reduction of the intracanal Enterococcus faecalis population was evaluated. Mandibular premolars with straight roots were selected. Teeth were decoronated and instrumented until 20 K file and irrigated with physiological saline. After sterilization by ethylene oxide gas, root canals were inoculated with Enterococcus faecalis. The specimens were randomly divided into five groups for canal instrumentation: Manual Nitiflex and Hero Shaper nickel titanium files, and rotary Hyflex CM, ProTaper Next, and K3XF nickel titanium files. Intracanal bacterial sampling was done before and after instrumentation. After serial dilution, samples were plated onto the Mitis Salivarius agar. The c.f.u. grown were counted, and log10 transformation was calculated. All instrumentation systems significantly reduced the intracanal bacterial population after root canal preparation. ProTaper Next was found to be significantly more effective than Hyflex CM and manual Nitiflex and Hero Shaper. However, ProTaper Next showed no significant difference with K3XF. Canal instrumentation by all the file systems significantly reduced the intracanal Enterococcus faecalis counts. ProTaper Next was found to be most effective in reducing the number of bacteria than other rotary or hand instruments. © 2014 Wiley Publishing Asia Pty Ltd.

Sanitization of an Automatic Reverse-Osmosis Watering System: Removal of a Clinically Significant Biofilm

PubMed Central

Molk, Denise M; Karr-May, Charlene L; Trang, Elaine D; Sanders, George E

2013-01-01

During environmental monitoring of our institution's rodent watering systems, one vivarium was found to have high bacterial loads in the reverse-osmosis (RO) automatic water system. These findings prompted evaluation of the entire RO water production and distribution system. Investigation revealed insufficient rack and RO system sanitization, leading to heavy biofilm accumulation within the system. Approximately 2 wk after discovery in the water system, one of the bacterial organisms isolated in the water supply, Sphingomonas paucimobilis, was isolated from a peritoneal abscess of a severely immunodeficient B6.Cg-Slc11a1r Rag1tm1Mom/Cwi mouse housed in the same vivarium, suggesting that rodents drinking from this system were being exposed randomly to fragments of biofilm. Plans were developed to sanitize the entire system. Hypercholorination was used first, followed by treatment with a combination of peracetic acid and hydrogen peroxide. Between system sanitizations, a low-level chlorine infusion was added to the system as a biocide. Heterotrophic plate counts and bacterial isolation were performed on water samples obtained before and after sanitization procedures. We here discuss the process of identifying and correcting this important water-quality issue. PMID:23562105

Adhesion of Asaia bogorensis to Glass and Polystyrene in the Presence of Cranberry Juice.

PubMed

Antolak, Hubert; Kregiel, Dorota; Czyzowska, Agata

2015-06-01

The aim of the study was to evaluate the adhesion abilities of the acetic acid bacterium Asaia bogorensis to glass and polystyrene in the presence of American cranberry (Vaccinium macrocarpon) juice. The strain of A. bogorensis used was isolated from spoiled commercial fruit-flavored drinking water. The cranberry juice was analyzed for polyphenols, organic acids, and carbohydrates using high-performance liquid chromatography and liquid chromatography-mass spectrometry techniques. The adhesive abilities of bacterial cells in culture medium supplemented with cranberry juice were determined using luminometry and microscopy. The viability of adhered and planktonic bacterial cells was determined by the plate count method, and the relative adhesion coefficient was calculated. This strain of A. bogorensis was characterized by strong adhesion properties that were dependent upon the type of surface. The highest level of cell adhesion was found on the polystyrene. However, in the presence of 10% cranberry juice, attachment of bacterial cells was three times lower. Chemical analysis of juice revealed the presence of sugars, organic acids, and anthocyanins, which were identified as galactosides, glucosides, and arabinosides of cyanidin and peonidin. A-type proanthocyanidins responsible for the antiadhesion properties of V. macrocarpon also were detected.

Regulation of Glucose Utilization by Estradiol in Breast Cancer

DTIC Science & Technology

2014-10-01

counts/min) weremeasured using 350 l of lysate. Protein concentration was determined using the BCA assay according to the manufacturer’s...instructions and measured on a Powerwave XS plate reader (Biotek). Counts were normalized to protein concentration. Glycolysis Assay—MCF-7 cells growing in 6...determined using the BCA assay according to the manufacturer’s instructions and mea- sured on a Powerwave XS plate reader. Counts were normal- ized to

Optimization, validation, and application of a real-time PCR protocol for quantification of viable bacterial cells in municipal sewage sludge and biosolids using reporter genes and Escherichia coli.

PubMed

van Frankenhuyzen, Jessica K; Trevors, Jack T; Flemming, Cecily A; Lee, Hung; Habash, Marc B

2013-11-01

Biosolids result from treatment of sewage sludge to meet jurisdictional standards, including pathogen reduction. Once government regulations are met, materials can be applied to agricultural lands. Culture-based methods are used to enumerate pathogen indicator microorganisms but may underestimate cell densities, which is partly due to bacteria existing in a viable but non-culturable physiological state. Viable indicators can also be quantified by realtime polymerase chain reaction (qPCR) used with propidium monoazide (PMA), a dye that inhibits amplification of DNA found extracellularly or in dead cells. The objectives of this study were to test an optimized PMA-qPCR method for viable pathogen detection in wastewater solids and to validate it by comparing results to data obtained by conventional plating. Reporter genes from genetically marked Pseudomonas sp. UG14Lr and Agrobacterium tumefaciens 542 cells were spiked into samples of primary sludge, and anaerobically digested and Lystek-treated biosolids as cell-free DNA, dead cells, viable cells, and mixtures of live and dead cells, followed by DNA extraction with and without PMA, and qPCR. The protocol was then used for Escherichia coli quantification in the three matrices, and results compared to plate counts. PMA-qPCR selectively detected viable cells, while inhibiting signals from cell-free DNA and DNA found in membrane-compromised cells. PMA-qPCR detected 0.5-1 log unit more viable E. coli cells in both primary solids and dewatered biosolids than plate counts. No viable E. coli was found in Lystek-treated biosolids. These data suggest PMA-qPCR may more accurately estimate pathogen cell numbers than traditional culture methods.

Evaluation of propidium monoazide real-time PCR for enumeration of probiotic lactobacilli microencapsulated in calcium alginate beads.

PubMed

Oketič, K; Matijašić, B Bogovič; Obermajer, T; Radulović, Z; Lević, S; Mirković, N; Nedović, V

2015-01-01

The aim of the study was to evaluate real-time PCR coupled with propidium monoazide (PMA) treatment for enumeration of microencapsulated probiotic lactobacilli microencapsulated in calcium alginate beads. Lactobacillus gasseri K7 (CCM 7710) and Lactobacillus delbrueckii subsp. bulgaricus (CCM 7712) were analysed by plate counting and PMA real-time PCR during storage at 4 °C for 90 days. PMA was effective in preventing PCR amplification of the target sequences of DNA released from heat-compromised bacteria. The values obtained by real-time PCR of non-treated samples were in general higher than those obtained by real-time PCR of PMA-treated samples or by plate counting, indicating the presence of sub-lethally injured cells. This study shows that plate count could not be completely replaced by culture independent method PMA real-time PCR for enumeration of probiotics, but may rather complement the well-established plate counting, providing useful information about the ratio of compromised bacteria in the samples.

Analysis of bacterial community shifts in the gastrointestinal tract of pigs fed diets supplemented with β-glucan from Laminaria digitata, Laminaria hyperborea and Saccharomyces cerevisiae.

PubMed

Murphy, P; Dal Bello, F; O'Doherty, J; Arendt, E K; Sweeney, T; Coffey, A

2013-07-01

This study was designed to evaluate the effects of algal and yeast β-glucans on the porcine gastrointestinal microbiota, specifically the community of Lactobacillus, Bifidobacterium and coliforms. A total of 48 pigs were fed four diets over a 28-day period to determine the effect that each had on these communities. The control diet consisted of wheat and soya bean meal. The remaining three diets contained wheat and soya bean meal supplemented with β-glucan at 250 g/tonne from Laminaria digitata, Laminaria hyperborea or Saccharomyces cerevisiae. Faecal samples were collected from animals before feeding each diet and after the feeding period. The animals were slaughtered the following day and samples were collected from the stomach, ileum, caecum, proximal colon and distal colon. Alterations in Lactobacillus in the gastrointestinal tract (GIT) were analysed using denaturing gradient gel electrophoresis (DGGE) profiles generated by group-specific 16S rRNA gene PCR amplicons. Plate count analysis was also performed to quantify total coliforms. DGGE profiles indicated that all β-glucan diets provoked the emergence of a richer community of Lactobacillus. The richest community of lactobacilli emerged after feeding L. digitata (LD β-glucan). Plate count analysis revealed that the L. hyperborea (LH β-glucan) diet had a statistically significant effect on the coliform counts in the proximal colon in comparison with the control diet. β-glucan from L. digitata and S. cerevisiae also generally reduced coliforms but to a lesser extent. Nevertheless, the β-glucan diets did not significantly reduce levels of Lactobacillus or Bifidobacterium. DGGE analysis of GIT samples indicated that the three β-glucan diets generally promoted the establishment of a more varied range of Lactobacillus species in the caecum, proximal and distal colon. The LH β-glucan had the most profound reducing effect on coliform counts when compared with the control diet and diets supplemented with L. digitata and S. cerevisiae β-glucans.

Effects of Extended Freezer Storage on the Integrity of Human Milk.

PubMed

Ahrabi, Ali Faraghi; Handa, Deepali; Codipilly, Champa N; Shah, Syed; Williams, Janet E; McGuire, Mark A; Potak, Debra; Aharon, Grace Golda; Schanler, Richard J

2016-10-01

To examine the integrity (pH, bacterial counts, host defense factors, nutrient contents, and osmolality) of freshly expressed and previously refrigerated human milk subjected to long-term freezer storage. Mothers donated 100 mL of freshly expressed milk. Samples were divided into baseline, storage at -20°C (fresh frozen) for 1, 3, 6, and 9 months, and prior storage at +4°C for 72 hours (refrigerated frozen) before storage at -20°C for 1 to 9 months. Samples were analyzed for pH, total bacterial colony count, gram-positive and gram-negative colony counts, and concentrations of total protein, fat, nonesterified fatty acids, lactoferrin, secretory IgA, and osmolality. Milk pH, total bacterial colony count, and Gram-positive colony counts decreased significantly with freezer storage (P < .001); bacterial counts decreased most rapidly in the refrigerated frozen group. The gram-negative colony count decreased significantly over time (P < .001). Nonesterified fatty acid concentrations increased significantly with time in storage (P < .001). Freezing for up to 9 months did not affect total protein, fat, lactoferrin, secretory IgA, or osmolality in either group. Freezer storage of human milk for 9 months at -20°C is associated with decreasing pH and bacterial counts, but preservation of key macronutrients and immunoactive components, with or without prior refrigeration for 72 hours. These data support current guidelines for freezer storage of human milk for up to 9 months for both freshly expressed and refrigerated milk. Copyright © 2016 Elsevier Inc. All rights reserved.

Effects of dark storage and retail display on beef chuck and round muscles enhanced with ammonium hydroxide, salt, and carbon monoxide.

PubMed

Hamling, A E; Jenschke, B E; Calkins, C R

2008-04-01

The objective of this study was to determine the retail shelf stability of beef chuck and round muscles enhanced with ammonium hydroxide, salt, and carbon monoxide. A split plot design was used for each of 3 muscles [triceps brachii (TB), biceps femoris (BF), and rectus femoris (RF)] with 2 treatments (0 and 20% pump), 3 dark storage periods (1, 2, and 3 wk), and 3 replications in the whole plot and retail display period as the split plot. There were a total of 12 subprimals per treatment per dark storage period (n = 72 each). Individual steaks were cut to a thickness of 2.54 cm and packaged in a modified-atmosphere package (MAP). The TB was packaged in a high-oxygen MAP (80% oxygen, 20% carbon dioxide). The BF and RF were packaged in a low-oxygen MAP (100% carbon dioxide). At the completion of each dark storage period, steaks were subjected to 7 d of simulated retail display. Steaks were used for objective and subjective color measurements, total plate counts, and determination of retail purge and oxidation. For all muscles, total plate counts were always numerically greater in injected steaks. Triceps brachii steaks held in dark storage for 3 wk and displayed at retail for 4 or more days all exceeded 10(7) log of cfu/cm(2) for aerobic plate count. Biceps femoris and RF steaks packaged in a low-oxygen MAP had much lower bacterial counts, with levels below 4.2 log of cfu/cm(2), even after 7 d of retail display. Oxidation values for the TB were extremely high (ranging from 12.3 to 26.6), whereas the BF and RF had values that were much lower (< or =1.0 mg of malonaldehyde/kg of muscle), likely due to the oxidation occurring in a high-oxygen MAP for the TB. Enhanced TB steaks proved to have greater color stability (less discoloration) than nonenhanced TB steaks. In addition, the BF and RF (low-oxygen MAP) steaks had better color stability (more stable redness values) than TB (high-oxygen MAP) steaks, although TB steaks initially exhibited a brighter red color. Retail display life was enhanced by packaging in 100% carbon dioxide, and enhanced steaks exhibited greater color stability in retail display than control steaks.

24-hour evaluation of dental plaque bacteria and halitosis after consumption of a single placebo or dental treat by dogs.

PubMed

Jeusette, Isabelle C; Román, Aurora Mateo; Torre, Celina; Crusafont, Josep; Sánchez, Nuria; Sánchez, Maria C; Pérez-Salcedo, Leire; Herrera, David

2016-06-01

OBJECTIVE To determine whether consumption of a single dental treat with specific mechanical properties and active ingredients would provide a 24-hour effect on dental plaque bacteria and halitosis in dogs. ANIMALS 10 dogs of various breeds from a privately owned colony that had received routine dental scaling and polishing 4 weeks before the study began. PROCEDURES Dogs were randomly assigned to receive 1 placebo or dental treat first. A 4-week washout period was provided, and then dogs received the opposite treatment. Oral plaque and breath samples were collected before and 0.5, 3, 12, and 24 hours after treat consumption. Volatile sulfur compounds (VSCs) concentration was measured in breath samples. Total aerobic, total anaerobic, Porphyromonas gulae, Prevotella intermedia-like, Tannerella forsythia, and Fusobacterium nucleatum bacterial counts (measured via bacterial culture) and total live bacterial counts, total live and dead bacterial counts, and bacterial vitality (measured via quantitative real-time PCR assay) were assessed in plaque samples. RESULTS Compared with placebo treat consumption, dental treat consumption resulted in a significant decrease in breath VSCs concentration and all plaque bacterial counts, without an effect on bacterial vitality. Effects of the dental treat versus the placebo treat persisted for 12 hours for several bacterial counts and for 24 hours for breath VSCs concentration. CONCLUSIONS AND CLINICAL RELEVANCE Although clinical benefits should be investigated in larger scale, longer-term studies, results of this study suggested that feeding the evaluated dental treat may help to decrease oral bacterial growth in dogs for 12 hours and oral malodor for 24 hours. A feeding interval of 12 hours is therefore recommended.

Bacterial transport in heterogeneous porous media: Observations from laboratory experiments

NASA Astrophysics Data System (ADS)

Silliman, S. E.; Dunlap, R.; Fletcher, M.; Schneegurt, M. A.

2001-11-01

Transport of bacteria through heterogeneous porous media was investigated in small-scale columns packed with sand and in a tank designed to allow the hydraulic conductivity to vary as a two-dimensional, lognormally distributed, second-order stationary, exponentially correlated random field. The bacteria were Pseudomonas ftuorescens R8, a strain demonstrating appreciable attachment to surfaces, and strain Ml, a transposon mutant of strain R8 with reduced attachment ability. In bench top, sand-filled columns, transport was determined by measuring intensity of fluorescence of stained cells in the effluent or by measuring radiolabeled cells that were retained in the sand columns. Results demonstrated that strain Ml was transported more efficiently than strain R8 through columns packed with either a homogeneous silica sand or a more heterogeneous sand with iron oxide coatings. Two experiments conducted in the tank involved monitoring transport of bacteria to wells via sampling from wells and sample ports in the tank. Bacterial numbers were determined by direct plate count. At the end of the first experiment, the distribution of the bacteria in the sediment was determined by destructive sampling and plating. The two experiments produced bacterial breakthrough curves that were quite similar even though the similarity between the two porous media was limited to first- and second-order statistical moments. This result appears consistent with the concept of large-scale, average behavior such as has been observed for the transport of conservative chemical tracers. The transported bacteria arrived simultaneously with a conservative chemical tracer (although at significantly lower normalized concentration than the tracer). However, the bacterial breakthrough curves showed significant late time tailing. The concentrations of bacteria attached to the sediment surfaces showed considerably more spatial variation than did the concentrations of bacteria in the fluid phase. This contrast between behavior in the fluid phase and on the solids is consistent with field observations by other authors and initial modeling of these heterogeneous media.

Evaluation of potassium-clavulanate-supplemented modified charcoal-cefoperazone-deoxycholate agar for enumeration of Campylobacter in chicken carcass rinse.

PubMed

Chon, Jung-Whan; Kim, Hong-Seok; Kim, Hyunsook; Oh, Deog-Hwan; Seo, Kun-Ho

2014-05-01

Potassium-clavulanate-supplemented modified charcoal-cefoperazone-deoxycholate agar (C-mCCDA) that was described in our previous study was compared with original mCCDA for the enumeration of Campylobacter in pure culture and chicken carcass rinse. The quantitative detection of viable Campylobacter cells from a pure culture, plated on C-mCCDA, is statistically similar (P > 0.05) to mCCDA. In total, 120 chickens were rinsed using 400 mL buffered peptone water. The rinses were inoculated onto C-mCCDA and mCCDA followed by incubation at 42 °C for 48 h. There was no statistical difference between C-mCCDA (45 of 120 plates; mean count, 145.5 CFU/mL) and normal mCCDA (46 of 120 plates; mean count, 160.8 CFU/mL) in the isolation rate and recovery of Campylobacter (P > 0.05) from chicken carcass rinse. The Pearson correlation coefficient value for the number of Campylobacter cells recovered in the 2 media was 0.942. However, the selectivity was much better on C-mCCDA than on mCCDA plates (P < 0.05). Significantly fewer C-mCCDA plates (33 out of 120 plates; mean count, 1.9 CFU/mL) were contaminated with non-Campylobacter cells than the normal mCCDA plates (67 out of 120 plates; mean count, 27.1 CFU/mL). The C-mCCDA may provide improved results for enumeration of Campylobacter in chicken meat alternative to mCCDA with its increased selectivity the modified agar possess. © 2014 Institute of Food Technologists®

Isolation and characterisation of phosphate solubilising microorganisms from the cold desert habitat of Salix alba Linn. in trans Himalayan region of Himachal Pradesh.

PubMed

Chatli, Anshu S; Beri, Viraj; Sidhu, B S

2008-06-01

Phosphate solubilising microorganisms (PSM) (bacteria and fungi) associated with Salix alba Linn. from Lahaul and Spiti valleys of Himachal Pradesh were isolated on Pikovskaya (PVK), modified Pikovskaya (MPVK) and National Botanical Research Institute agar (NBRIP) media by spread plating. The viable colony count of P-solubilising bacteria (PSB) and fungi (PSF) was higher in rhizosphere than that of non-rhizosphere. The frequency of PSM was highest on MPVK followed by NBRIP and PVK agar. The maximum proportion of PSM out of total bacterial and fungal count was found in upper Keylong while the least in Rong Tong. The PSB frequently were Gram-positive, endosporeforming, motile rods and belonged to Bacillus sp. The PSF mainly belonged to Penicillium sp., Aspergillus fumigatus, A. niger, A. spp. and non-sporulating sterile. Amongst the isolates with high efficiency for tricalcium phosphate (TCP) solubilisation, seven bacterial and seven fungal isolates dissolved higher amount of P from North Carolina rock phosphate (NCRP) than Mussoorie rock phosphate (MRP) and Udaipur rock phosphate (URP). However, the organisms solubilised higher-P in NBRIP broth than PVK broth. SBC5 (Bacillus sp.) and SBC7 (Bacillus sp.) bacterial isolates exhibited maximun P solubilisation (40 and 33 μg ml(-1) respectively) whereas FC28 (Penicillium sp.) isolate (52.3 μg ml(-1)) amongst fungi while solubilising URP. The amount of P solubilised was positively correlated with the decrease in pH of medium. SBC5 (Bacillus sp.), SBC7 (Bacillus sp.) and SBC4 (Micrococcus) decreased the pH of medium from 6.8 to 6.08 while FC28 (Penicillium sp.) and FC39 (Penicillium sp.) isolates of fungi recorded maximum decrease in pH of medium from 6.8 to 5.96 in NBRIP broth.

Preliminary Efficacy Testing of the Disinfectant MicrobeCare XLP for Potential Use in Military Operational Environments.

PubMed

Madden, Jonathan F; Henrichs, Lori; Ervin, Mark D; Lospinoso, Joshua; Beachkofsky, Thomas M; Hardin, Carolyn A

2018-04-04

A safe, easy-to-use, permanently bonded antiseptic that does not require post-exposure bioload reduction but maintains effectiveness over time would have far-reaching implications across multiple industries. Health care is one such arena, particularly in austere military settings where resources are at a premium. MicrobeCare XLP (MicrobeCare, Buffalo Grove, IL, USA) is a commercially available spray-on agent that is advertised to covalently bond to surfaces and provide a long-lasting antimicrobial coating inhospitable to >99.99% of surface microorganisms. A pilot study was devised to gather baseline data regarding product efficacy and laboratory parameters before consideration of extended investigations and military utilization. The product manufacturer recommends bioload reductions before product application, following product application, and after each pathogenic exposure. To investigate the product's efficacy in circumstances more closely simulating a military operational setting in which post-pathogenic exposure bioload reduction would not be possible, this step was deliberately excluded from the test sequences. Using autoclaved surgical forceps, growth of Staphylococcus aureus and Acinetobacter baumannii was evaluated in a controlled manner under multiple conditions. Test variations included duration of submersion in the MicrobeCare XLP solution and air-drying and a second autoclave sterilization. Control and treated forceps were exposed to a bacterial suspension and air-dried before being submerged in sterile saline and vortex mixed. The saline solution was serially diluted and plated on tryptic soy agar (TSA) II plates. Plates were incubated for 24 h and bacterial colony-forming units (CFU)/mL were counted. Statistical significance was defined according to the American Society for Testing and Materials (ASTM) International passing criteria of 3 Log10 or 99.9% reduction of microorganisms. Additionally, p-values were calculated using two-tailed unpaired two-sample t-tests with unequal variance with a threshold of 0.05. In the S. aureus tests, none of the reduction calculations met the ASTM International passing criteria. In addition, the difference between the means of the colony counts in the MicrobeCare XLP-treated forceps and untreated control forceps was not statistically significant (p-value 0.109). Conversely, in the A. baumannii tests, each of the percent reduction calculations met the ASTM International passing criteria; the difference between the means of the colony counts in the treatment and control groups was statistically significant (p-value 0.008). In these independent tests, MicrobeCare XLP effectively prevented growth of A. baumannii but had unpredictable results suppressing S. aureus. These results may relate to inherent properties of the bacteria or autoclave exposure, although the manufacturer asserts that the coating withstands such degradation. Additional testing could be performed using a broader range of microorganisms and exposure to varying conditions including other sterilization methods.

Indoor air bacterial load and antibiotic susceptibility pattern of isolates in operating rooms and surgical wards at jimma university specialized hospital, southwest ethiopia.

PubMed

Genet, Chalachew; Kibru, Gebre; Tsegaye, Wondewosen

2011-03-01

Surgical site infection is the second most common health care associated infection. One of the risk factors for such infection is bacterial contamination of operating rooms' and surgical wards' indoor air. In view of that, the microbiological quality of air can be considered as a mirror of the hygienic condition of these rooms. Thus, the objective of this study was to determine the bacterial load and antibiotic susceptibility pattern of isolates in operating rooms' and surgical wards' indoor air of Jimma University Specialized Hospital. A cross sectional study was conducted to measure indoor air microbial quality of operating rooms and surgical wards from October to January 2009/2010 on 108 indoor air samples collected in twelve rounds using purposive sampling technique by Settle Plate Method (Passive Air Sampling following 1/1/1 Schedule). Sample processing and antimicrobial susceptibility testing were done following standard bacteriological techniques. The data was analyzed using SPSS version 16 and interpreted according to scientifically determined baseline values initially suggested by Fisher. The mean aerobic colony counts obtained in OR-1(46cfu/hr) and OR-2(28cfu/hr) was far beyond the set 5-8cfu/hr acceptable standards for passive room. Similarly the highest mean aerobic colony counts of 465cfu/hr and 461cfu/hr were observed in Female room-1 and room-2 respectively when compared to the acceptable range of 250-450cfu/hr. In this study only 3 isolates of S. pyogenes and 48 isolates of S. aureus were identified. Over 66% of S. aureus was identified in Critical Zone of Operating rooms. All isolates of S. aureus showed 100% and 82.8% resistance to methicillin and ampicillin respectively. Higher degree of aerobic bacterial load was measured from operating rooms' and surgical wards' indoor air. Reducing foot trafficking, improving the ventilation system and routine cleaning has to be made to maintain the aerobic bacteria load with in optimal level.

Relative Numbers of Certain Microbial Groups Present in Compost Used for Mushroom (Agaricus bisporus) Propagation

PubMed Central

Fordyce, C.

1970-01-01

The relative numbers of microorganisms associated with compost during mushroom production were studied by the dilution plate method. Thermophilic actinomycetes and fungi were isolated with a very high frequency early in the growing season. Although numbers of thermophilic bacteria diminished slowly during the season, the thermophilic fungi and actinomycetes diminished rapidly with the latter disappearing after 6 weeks. Mesophilic fungi other than Agaricus or Trichoderma remained relatively stable throughout the growing period. Agaricus could be isolated between the first and third break. Trichoderma became dominant after the fourth break. The mesophilic bacterial counts diminished during the most productive portion of the mushroom cropping season and then increased to much higher numbers toward the end of the season. PMID:5529631

New method for estimating bacterial cell abundances in natural samples by use of sublimation

NASA Technical Reports Server (NTRS)

Glavin, Daniel P.; Cleaves, H. James; Schubert, Michael; Aubrey, Andrew; Bada, Jeffrey L.

2004-01-01

We have developed a new method based on the sublimation of adenine from Escherichia coli to estimate bacterial cell counts in natural samples. To demonstrate this technique, several types of natural samples, including beach sand, seawater, deep-sea sediment, and two soil samples from the Atacama Desert, were heated to a temperature of 500 degrees C for several seconds under reduced pressure. The sublimate was collected on a cold finger, and the amount of adenine released from the samples was then determined by high-performance liquid chromatography with UV absorbance detection. Based on the total amount of adenine recovered from DNA and RNA in these samples, we estimated bacterial cell counts ranging from approximately 10(5) to 10(9) E. coli cell equivalents per gram. For most of these samples, the sublimation-based cell counts were in agreement with total bacterial counts obtained by traditional DAPI (4,6-diamidino-2-phenylindole) staining.

Multiple Heavy Metal Tolerance of Soil Bacterial Communities and Its Measurement by a Thymidine Incorporation Technique

PubMed Central

Díaz-Raviña, Montserrat; Bååth, Erland; Frostegård, Åsa

1994-01-01

A thymidine incorporation technique was used to determine the tolerance of a soil bacterial community to Cu, Cd, Zn, Ni, and Pb. An agricultural soil was artificially contaminated in our laboratory with individual metals at three different concentrations, and the results were compared with the results obtained by using the plate count technique. Thymidine incorporation was found to be a simple and rapid method for measuring tolerance. Data obtained by this technique were very reproducible. A linear relationship was found between changes in community tolerance levels obtained by the thymidine incorporation and plate count techniques (r = 0.732, P < 0.001). An increase in tolerance to the metal added to soil was observed for the bacterial community obtained from each polluted soil compared with the community obtained from unpolluted soil. The only exception was when Pb was added; no indication of Pb tolerance was found. An increase in the tolerance to metals other than the metal originally added to soil was also observed, indicating that there was multiple heavy metal tolerance at the community level. Thus, Cu pollution, in addition to increasing tolerance to Cu, also induced tolerance to Zn, Cd, and Ni. Zn and Cd pollution increased community tolerance to all five metals. Ni amendment increased tolerance to Ni the most but also increased community tolerance to Zn and, to lesser degrees, increased community tolerance to Pb and Cd. In soils polluted with Pb increased tolerance to other metals was found in the following order: Ni > Cd > Zn > Cu. We found significant positive relationships between changes in Cd, Zn, and Pb tolerance and, to a lesser degree, between changes in Pb and Ni tolerance when all metals and amendment levels were compared. The magnitude of the increase in heavy metal tolerance was found to be linearly related to the logarithm of the metal concentration added to the soil. Threshold tolerance concentrations were estimated from these linear relationships, and changes in tolerance could be detected at levels of soil contamination similar to those reported previously to result in changes in the phospholipid fatty acid pattern (Å. Frostegård, A. Tunlid, and E. Bååth, Appl. Environ. Microbiol. 59: 3605-3617, 1993). PMID:16349314

Influence of phosphate and disinfection on the composition of biofilms produced from drinking water, as measured by fluorescence in situ hybridization.

PubMed

Batté, M; Mathieu, L; Laurent, P; Prévost, M

2003-12-01

Biofilms were grown in annular reactors supplied with drinking water enriched with 235 microg C/L. Changes in the biofilms with ageing, disinfection, and phosphate treatment were monitored using fluorescence in situ hybridization. EUB338, BET42a, GAM42a, and ALF1b probes were used to target most bacteria and the alpha (alpha), beta (beta), and gamma (gamma) subclasses of Proteobacteria, respectively. The stability of biofilm composition was checked after the onset of colonization between T = 42 days and T = 113 days. From 56.0% to 75.9% of the cells detected through total direct counts with DAPI (4'-6-diamidino-2-phenylindole) were also detected with the EUB338 probe, which targets the 16S rRNA of most bacteria. Among these cells, 16.9%-24.7% were targeted with the BET42a probe, 1.8%-18.3% with the ALF1b probe, and <2.5% with the GAM42a probe. Phosphate treatment induced a significant enhancement to the proportion of gamma-Proteobacteria (detected with the GAM42a probe), a group that contains many health-related bacteria. Disinfection with monochloramine for 1 month or chlorine for 3 days induced a reduction in the percentage of DAPI-stained cells that hybridized with the EUB338 probe (as expressed by percentages of EUB338 counts/DAPI) and with any of the ALF1b, BET42a, and GAM42a probes. The percentage of cells detected by any of the three probes (ALF1b+BET42a+GAM42a) tended to decrease, and reached in total less than 30% of the EUB338-hybridized cells. Disinfection with chlorine for 7 days induced a reverse shift; an increase in the percentage of EUB338 counts targeted by any of these three probes was noted, which reached up to 87%. However, it should be noted that the global bacterial densities (heterotrophic plate counts and total direct counts) tended to decrease over the duration of the experiment. Therefore, those bacteria that could be considered to resist 7 days of chlorination constituted a small part of the initial biofilm community, up to the point at which the other bacterial groups were destroyed by chlorination. The results suggest that there were variations in the kinetics of inactivation by disinfectant, depending on the bacterial populations involved.

The utility of biomarkers in differentiating bacterial from non-bacterial lower respiratory tract infection in hospitalized children: difference of the diagnostic performance between acute pneumonia and bronchitis.

PubMed

Hoshina, Takayuki; Nanishi, Etsuro; Kanno, Shunsuke; Nishio, Hisanori; Kusuhara, Koichi; Hara, Toshiro

2014-10-01

The aim of this study is to investigate the utility of several biomarkers in differentiating bacterial community-acquired lower respiratory tract infection (CA-LRTI) from non-bacterial CA-LRTI in children and the difference of their diagnostic performance between pneumonia and bronchitis. A retrospective cohort study composed of 108 pediatric patients hospitalized for CA-LRTI was performed during 2010-2013. Based on the findings of chest X-ray and sputum samples, patients were divided into 4 categories, group of bacterial pneumonia or bronchitis, and non-bacterial (viral or etiology-unknown) pneumonia or bronchitis. Peripheral white blood cell and neutrophil counts, and serum C-reactive protein (CRP) and procalcitonin (PCT) levels were compared among the 4 groups. Finally, 54 patients were the subject of this study. In the patients with pneumonia, serum CRP and PCT levels were significantly elevated in the group of bacterial pneumonia (CRP: p = 0.02, PCT: p = 0.0008). The area under the receiver operating characteristic curve for PCT for distinguishing between bacterial and non-bacterial pneumonia was the largest, and sensitivity, specificity, positive predictive value and negative predictive value of PCT were best among 4 markers. On the other hand, in the patients with bronchitis, neutrophil count was significantly decreased in non-bacterial bronchitis whereas no significant differences of WBC count, CRP level or PCT level were seen. In conclusion, PCT was the most useful marker to differentiate bacterial pneumonia whereas neutrophil count contributed most to the discrimination of bacterial bronchitis. The diagnostic performance of biomarkers may be different between pneumonia and bronchitis. Copyright © 2014. Published by Elsevier Ltd.

Effects of open drainage ditch design on bacterial and fungal communities of cold waterlogged paddy soils

PubMed Central

Qiu, Shanlian; Wang, MK; Wang, Fei; Chen, Jichen; Li, Xiaoyan; Li, Qinghua; Lin, Cheng; Lin, Xinjian

2013-01-01

A field experiment established in 1980 was conducted to evaluate the effects of open drainage ditch applied for water removal on bacterial and fungal communities of cold waterlogged paddy soils in 2011. In this experiment, traditional plate counting and temperature gradient gel electrophoresis were employed to characterize the abundance and diversity of soil bacterial and fungal communities. Four different distances from the open drainage ditch, 5, 15, 25 and 75 m with different degrees of drainage were designed for this study. Maximum populations of culturable aerobic bacteria and fungi were at 15-m distance while minimum populations were at 75-m distance. Significant differences (p < 0.05) in fungal populations were observed at all distances from open drainage ditch. The highest diversity of the bacterial community was found at a distance of 25 m, while that of the fungal community was observed at a distance of 5 m. Sequencing of excised TGGE bands indicated that the dominant bacteria at 75-m distance belonged to anaerobic or microaerobic bacteria. Relationships between microbial characteristics and soil physicochemical properties indicated that soil pH and available nitrogen contents were key factors controlling the abundance of culturable aerobic bacteria and fungi, while soil water capacity also affected the diversity of fungal community. These findings can provide the references for better design and advanced management of the drainage ditches in cold waterlogged paddy soils. PMID:24516468

An investigation of total bacterial communities, culturable antibiotic-resistant bacterial communities and integrons in the river water environments of Taipei city.

PubMed

Yang, Chu-Wen; Chang, Yi-Tang; Chao, Wei-Liang; Shiung, Iau-Iun; Lin, Han-Sheng; Chen, Hsuan; Ho, Szu-Han; Lu, Min-Jheng; Lee, Pin-Hsuan; Fan, Shao-Ning

2014-07-30

The intensive use of antibiotics may accelerate the development of antibiotic-resistant bacteria (ARB). The global geographical distribution of environmental ARB has been indicated by many studies. However, the ARB in the water environments of Taiwan has not been extensively investigated. The objective of this study was to investigate the communities of ARB in Huanghsi Stream, which presents a natural acidic (pH 4) water environment. Waishuanghsi Stream provides a neutral (pH 7) water environment and was thus also monitored to allow comparison. The plate counts of culturable bacteria in eight antibiotics indicate that the numbers of culturable carbenicillin- and vancomycin-resistant bacteria in both Huanghsi and Waishuanghsi Streams are greater than the numbers of culturable bacteria resistant to the other antibiotics tested. Using a 16S rDNA sequencing approach, both the antibiotic-resistant bacterial communities (culture-based) and the total bacterial communities (metagenome-based) in Waishuanghsi Stream exhibit a higher diversity than those in Huanghsi Stream were observed. Of the three classes of integron, only class I integrons were identified in Waishuanghsi Stream. Our results suggest that an acidic (pH 4) water environment may not only affect the community composition of antibiotic-resistant bacteria but also the horizontal gene transfer mediated by integrons. Copyright © 2013 Elsevier B.V. All rights reserved.

Action of Monomeric/Gemini Surfactants on Free Cells and Biofilm of Asaia lannensis.

PubMed

Koziróg, Anna; Kręgiel, Dorota; Brycki, Bogumił

2017-11-22

We investigated the biological activity of surfactants based on quaternary ammonium compounds: gemini surfactant hexamethylene-1,6-bis-( N,N -dimethyl- N -dodecylammonium bromide) (C6), synthesized by the reaction of N,N -dimethyl- N- dodecylamine with 1,6-dibromohexane, and its monomeric analogue dodecyltrimethylammonium bromide (DTAB). The experiments were performed with bacteria Asaia lannensis , a common spoilage in the beverage industry. The minimal inhibitory concentration (MIC) values were determined using the tube standard two-fold dilution method. The growth and adhesive properties of bacterial cells were studied in different culture media, and the cell viability was evaluated using plate count method. Both of the surfactants were effective against the bacterial strain, but the MIC of gemini compound was significantly lower. Both C6 and DTAB exhibited anti-adhesive abilities. Treatment with surfactants at or below MIC value decreased the number of bacterial cells that were able to form biofilm, however, the gemini surfactant was more effective. The used surfactants were also found to be able to eradicate mature biofilms. After 4 h of treatment with C6 surfactant at concentration 10 MIC, the number of bacterial cells was reduced by 91.8%. The results of this study suggest that the antibacterial activity of the gemini compound could make it an effective microbiocide against the spoilage bacteria Asaia sp. in both planktonic and biofilm stages.

Retreived bacteria from Noctiluca miliaris (green) bloom of the northeastern Arabian Sea

NASA Astrophysics Data System (ADS)

Basu, Subhajit; Matondkar, S. G. Prabhu; Furtado, Irene

2013-01-01

In recent years, seasonal blooms of the dinoflagellate Noctiluca miliaris have appeared in the open-waters of the northern Arabian Sea (NAS). This study provides the first characterization of bacteria from a seasonal bloom of green Noctiluca of NAS (20°N-17°N and 64°E-70°E), during the spring-inter-monsoon cruise of Sagar Sampada 253, in March 2007. Bacterial growth as assessed by most-probable number (MPN) and plate counts, revealed `variable-physiotypes' over a wide range of salinities (0%-25% w/v NaCl), pH levels (5-8.5), and organic nutrient strengths, in comparison to non-bloom waters. MPN indices of bacteria in surface waters of bloom stations *DWK and *PRB, corresponded to (3.08-4.41)×103 cells/mL at 3.5% NaCl (w/v), and (2.82-9.49)×102 cells/mL at 25% (w/v) NaCl in tryptone-yeast extract broth (TYE). Plate counts were (1.12-4)×106 CFU/mL at 0% (w/v) NaCl, (1.28-3.9)×106 CFU/mL at 3.5% (w/v) NaCl, and (0.4-7)×104 CFU/mL at 25% NaCl (w/v) on TYE. One-tenth-strength Zobell's gave (0.6-3.74)×105 CFU/mL at pH 5 to (3.58-7.5)×105 CFU/mL at pH 8.5. These bacteria were identified to the genera Bacillus, Cellulomonas, Staphylococcus, Planococcus, Dietzia, Virgibacillus, Micrococcus, Sporosarcinae, Leucobacter, and Halomonas. The identity of three strains (GUFBSS253N2, GUFBSS253N30, and GUFBSS253N84) was confirmed through 16S rDNA sequence homology as Bacillus cohnii, Bacillus flexus, and Bacillus cereus. The ˜2-3-fold higher plate counts of culturable bacteria from the open-waters of the NAS indicate that these bacteria could critically determine the biogeochemical dynamics of the bloom and its milieu. The role of these bacteria in sustaining/terminating the bloom is under evaluation.

Comparison of direct-plating and broth-enrichment culture methods for detection of potential bacterial pathogens in respiratory secretions.

PubMed

Kaur, Ravinder; Wischmeyer, Jareth; Morris, Matthew; Pichichero, Michael E

2017-11-01

We compared the recovery of potential respiratory bacterial pathogens and normal flora from nasopharyngeal specimens collected from children during health and at the onset of acute otitis media (AOM) by selective direct-plating and overnight broth-enrichment. Overall, 3442 nasal wash (NW) samples collected from young children were analysed from a 10-year prospective study. NWs were cultured by (1) direct-plating to TSAII/5 % sheep blood agar and chocolate agar plates and (2) overnight broth-enrichment in BacT/ALERT SA-broth followed by plating. Standard microbiology techniques were applied to identify three dominant respiratory bacterial pathogens: Streptococcus pneumoniae (Spn), Haemophilus influenzae (Hflu) and Moraxella catarrhalis (Mcat) as well as two common nasal flora, Staphylococcus aureus (SA) and alpha-haemolytic Streptococci (AHS).Results/Key findings. Direct-plating of NW resulted in isolation of Spn from 37.8 %, Hflu from 13.6 % and Mcat from 33.2 % of samples. In comparison, overnight broth-enrichment isolated fewer Spn (30.1 %), Hflu (6.2 %) and Mcat (16.2 %) (P<0.001-0.0001). Broth-enrichment resulted in significant increased isolation of SA (6.0 %) and AHS (30.1 %) (P<0.0001). Competition between bacterial species in broth when both species were detected by direct-plating was assessed, and it was found that SA and AHS out-competed other species during broth-enrichment when samples were collected from healthy children but not during AOM. In middle ear fluids (MEF) at the onset of AOM, broth-enrichment resulted in higher recovery of Spn (+10.4 %, P<0.001), Hflu (+4.4 %, P=0.39) and Mcat (+13.5 %, <0.001). Broth-enrichment significantly reduces the accurate detection of bacterial respiratory pathogens and increases identification of SA and AHS in NW. Broth-enrichment improves detection of bacterial respiratory pathogens in MEF samples.

Survey of bovine colostrum quality and hygiene on northern Victorian dairy farms.

PubMed

Phipps, A J; Beggs, D S; Murray, A J; Mansell, P D; Stevenson, M A; Pyman, M F

2016-11-01

One of the major challenges for dairy producers is to produce, harvest, and store high-quality colostrum and feed it to their replacement heifer calves. Limited published data are available in Australia regarding the relationship between colostrum management, hygiene, and quality. The objectives of this study were to investigate (1) the colostrum storage and handling practices carried out on farm; (2) the immunoglobulin concentration and bacterial composition of colostrum being fed to replacement dairy heifer calves; (3) the percentage of colostrum being fed to replacement dairy heifer calves that meet industry recommendations; and (4) risk factors for bacterial contamination of colostrum. The study was carried out on 24 dairy farms located near Rochester, Victoria, Australia. Two hundred forty colostrum samples were collected (10 samples per farm). Each farm harvested and stored first-milking colostrum under normal farm conditions. A 10-mL sample of the colostrum was collected in a sterile container immediately before feeding, and a Brix refractometer reading was taken. The samples were then frozen at -4°C and submitted for bacterial concentration analysis. Fifty-eight percent of colostrum samples met the recommended industry standard of a total plate count (TPC) of <100,000cfu/mL, and 94% of colostrum samples met the recommended industry standard of total coliform count (TCC) of 10,000cfu/mL. However, when all the current industry recommendations for TPC, TCC, and Brix refractometer percentage for colostrum quality were considered, only 23% of the samples met all standards. These findings demonstrate that a large number of calves were at risk of receiving colostrum of poor quality, with high bacterial loads that may have interfered with the acquisition of transfer of passive immunity and affected calf health. Further investigation is required to identify the farm-specific factors that may influence the level of bacterial contamination of colostrum. Recommendations as a result of this study include refrigeration of excess colostrum shortly (within 1h) after collection and thorough disinfection of the calf feeding apparatus before use. Copyright © 2016 American Dairy Science Association. Published by Elsevier Inc. All rights reserved.

The effect of storage temperature and duration on the microbial quality of bulk tank milk.

PubMed

O'Connell, A; Ruegg, P L; Jordan, K; O'Brien, B; Gleeson, D

2016-05-01

The dairy industry in Ireland is currently undergoing a period of expansion and, as a result, it is anticipated that milk may be stored in bulk tanks on-farm for periods greater than 48 h. The objective of this study was to investigate the effects of storage temperature and duration on microbial quality of bulk tank milk when fresh milk is added to the bulk tank twice daily. Bulk tank milk stored at 3 temperatures was sampled at 24-h intervals during storage periods of 0 to 96 h. Bulk tank milk samples were analyzed for total bacterial count (TBC), psychrotrophic bacterial count (PBC), laboratory pasteurization count (LPC), psychrotrophic-thermoduric bacterial count (PBC-LPC), proteolytic bacterial count, lipolytic bacterial count, presumptive Bacillus cereus, sulfite-reducing Clostridia (SRC), and SCC. The bulk tank milk temperature was set at each of 3 temperatures (2°C, 4°C, and 6°C) in each of 3 tanks on 2 occasions during two 6-wk periods. Period 1 was undertaken in August and September, when all cows were in mid lactation, and period 2 was undertaken in October and November, when all cows were in late lactation. None of the bulk tank bacterial counts except the proteolytic count were affected by lactation period. The proteolytic bacterial count was greater in period 2 than in period 1. The TBC and PBC of milk stored at 6°C increased as storage duration increased. The TBC did not increase with increasing storage duration when milk was stored at 2°C or 4°C but the PBC of milk stored at 4°C increased significantly between 0 and 96 h. The numbers of proteolytic and lipolytic bacteria, LPC, or PBC-LPC in bulk tank milk were not affected by temperature or duration of storage. Presumptive B. cereus were detected in 10% of all bulk tank milk samples taken over the two 6-wk periods, with similar proportions observed in both. In bulk tank milk samples, a greater incidence of SRC was observed in period 2 (20%) compared with period 1 (3%). Milk produced on-farm with minimal bacterial contamination can be successfully stored at 2°C and 4°C for up to 96h with little effect on its microbial quality. Copyright © 2016 American Dairy Science Association. Published by Elsevier Inc. All rights reserved.

A Hidden Pitfall in the Preparation of Agar Media Undermines Microorganism Cultivability

PubMed Central

Tanaka, Tomohiro; Kawasaki, Kosei; Daimon, Serina; Kitagawa, Wataru; Yamamoto, Kyosuke; Tamaki, Hideyuki; Tanaka, Michiko; Nakatsu, Cindy H.

2014-01-01

Microbiologists have been using agar growth medium for over 120 years. It revolutionized microbiology in the 1890s when microbiologists were seeking effective methods to isolate microorganisms, which led to the successful cultivation of microorganisms as single clones. But there has been a disparity between total cell counts and cultivable cell counts on plates, often referred to as the “great plate count anomaly,” that has long been a phenomenon that still remains unsolved. Here, we report that a common practice microbiologists have employed to prepare agar medium has a hidden pitfall: when phosphate was autoclaved together with agar to prepare solid growth media (PT medium), total colony counts were remarkably lower than those grown on agar plates in which phosphate and agar were separately autoclaved and mixed right before solidification (PS medium). We used a pure culture of Gemmatimonas aurantiaca T-27T and three representative sources of environmental samples, soil, sediment, and water, as inocula and compared colony counts between PT and PS agar plates. There were higher numbers of CFU on PS medium than on PT medium using G. aurantiaca or any of the environmental samples. Chemical analysis of PT agar plates suggested that hydrogen peroxide was contributing to growth inhibition. Comparison of 454 pyrosequences of the environmental samples to the isolates revealed that taxa grown on PS medium were more reflective of the original community structure than those grown on PT medium. Moreover, more hitherto-uncultivated microbes grew on PS than on PT medium. PMID:25281372

The physical and microbiological quality of chicken meat in the different type of enterprise poultry slaughterhouse: a case study in Karanganyar District

NASA Astrophysics Data System (ADS)

Hertanto, B. S.; Nurmalasari, C. D. A.; Nuhriawangsa, A. M. P.; Cahyadi, M.; Kartikasari, L. R.

2018-01-01

The aim of this study was to determine the physical and microbiological quality of chicken meat produced by the different type of enterprise slaughterhouse in Karanganyar District. The number of 20 poultry slaughterhouses was determined by convenience sampling method. The samples of chicken meat were randomly collected from medium enterprise poultry slaughterhouses (n=12) and small enterprise poultry slaughterhouses (n=8). A survey was carried out among poultry slaughterhouses in Karanganyar District. All the samples were subjected to physical quality consisted of pH test, texture, and color, while microbiological quality consisted of total plate count, microbial detection of Escherichia coli and Salmonella. The data were analyzed using descriptive quantitative analysis. The study showed that chicken meat in 6 small enterprise slaughterhouses and 11 medium enterprise slaughterhouses had normal pH of 5.81 - 6.3. Color and texture of chicken meats had relatively normal in both small and medium enterprise slaughterhouses. The total plate count of chicken meat showed in both small and medium enterprise slaughterhouses was <1x106 CFU/gr. The test of bacterial contamination showed that 3 of small and medium enterprise slaughterhouses were positively contaminated by Escherichia coli of >1x101 CFU/gr, and Salmonella was detected in 1 medium enterprise slaughterhouse. The overall results of the study suggest that the potential risk of chicken meat contamination depends on the processing of chicken meat in poultry slaughterhouses.

Effects of Bite Count Feedback from a Wearable Device and Goal Setting on Consumption in Young Adults.

PubMed

Jasper, Phillip W; James, Melva T; Hoover, Adam W; Muth, Eric R

2016-11-01

New technologies are emerging that may help individuals engage in healthier eating behaviors. One paradigm to test the efficacy of a technology is to determine its effect relative to environment cues that are known to cause individuals to overeat. The purpose of this work was to independently investigate two questions: How does the presence of a technology that provides bite count feedback alter eating behavior? and, How does the presence of a technology that provides bite count feedback paired with a goal alter eating behavior? Two studies investigated these research questions. The first study tested the effects of a large and small plate crossed with the presence or absence of a device that provided bite count feedback on intake. The second study tested the effects of a bite count goal with bite count feedback, again crossed with plate size, on intake. Both studies used a 2×2 between-subjects design. In the first study, 94 subjects (62 women aged 19.0±1.6 years with body mass index [BMI] 23.04±3.6) consumed lunch in a laboratory. The second study examined 99 subjects (56 women aged 18.5±1.5 years with BMI 22.73±2.70) under the same conditions. In both studies subjects consumed a single-course meal, using either a small or large plate. In the first study participants either wore or did not wear an automated bite counting device. In the second study all participants wore the bite counting device and were given either a low bite count goal (12 bites) or a high bite count goal (22 bites). Effect of plate size, feedback, and goal on consumption (grams) and number of bites taken were assessed using 2×2 analyses of variance. As adjunct measures, the effects of serving size, bite size (grams per bite), postmeal satiety, and satiety change were also assessed. In the first study there was a main effect of plate size on grams consumed and number of bites taken such that eating from a large plate led to greater consumption (P=0.001) and a greater number of bites (P=0.001). There was also a main effect of feedback on consumption and number of bites taken such that those who received feedback consumed less (P=0.011) and took fewer bites (P<0.001). In the second study there was a main effect of plate size on consumption such that those eating from a large plate consumed more (P=0.003) but did not take more bites. Further analysis revealed a main effect of goal on number of bites taken such that those who received the low goal took fewer bites (P<0.001) but did not consume less. Providing feedback on the number of bites taken from a wearable intake monitor can reduce overall intake during a single meal. Regarding the first research question, providing feedback significantly reduced intake in both plate size groups and reduced the overall number of bites taken. Regarding the second research question, participants were successful in eating to their goals. However, individuals in the low goal condition appeared to compensate for the restricted goal by taking larger bites, leading to comparable levels of consumption between the low and high goal groups. Hence, the interaction of technology with goals should be considered when introducing a health intervention. Copyright © 2016 Academy of Nutrition and Dietetics. Published by Elsevier Inc. All rights reserved.

[Endotoxin Contamination and Correlation with Other Water Quality Parameters of Groundwater from Self-Contained Wells in Beijing].

PubMed

Zhang, Can; Liu, Wen-jun; Ao, Lu; Shi, Yun; An, Dai-zhi; Liu, Zhi-ping

2015-12-01

A survey of endotoxin activity in groundwater from 14 self-contained wells in PLA units stationed in Beijing was conducted by the kinetic-turbid assay of Tachypleus Amebocyte Lysate (TAL). Bacteriological parameters, including total cell counts detected by flow cytometry, heterotrophic plate counts (HPC), standard plate counts and total coliforms were analyzed. Additionally, suspended particles, turbidity, dissolved organic carbon (DOC), and UV₂₅₄ were investigated. Total endotoxin activities ranged from 0. 15 to 13.20 EU · mL⁻¹, free endotoxin activities ranged from 0.10 to 5.29 EU · mL⁻¹ and bound endotoxin activities ranged from 0.01 to 8.60 EU · mL⁻¹. Most of the endotoxins in heavily contaminated groundwater existed as bound endotoxins. As for total endotoxins, the sequence of correlation coefficients with other parameters was total cell counts (r = 0.88 ) > HPC (r = 0.79) > DOC (r = 0.77) > UV₂₅₄ (r = 0.57) > total coliforms (r = 0.50) > standard plate counts (r = 0.49) = turbidity (r = 0. 49) > total particles (r = 0.41). The sequence of correlations of the bound endotoxins with other parameters was total cell counts (r = 0.81) > HPC (r = 0.66) > total coliforms (r = 0.65) > turbidity (r = 0.62) > total particles (r = 0.58) > standard plate counts (r = 0.22). Free endotoxins were correlated with DOC and UV₂₅₄, r = 0.58 and 0.26, respectively. Result showed free endotoxins had a higher correlation with DOC, and a lower correlation with UV₂₅₄.

Survival of bioluminescent Listeria monocytogenes and Escherichia coli O157:H7 in soft cheeses.

PubMed

Ramsaran, H; Chen, J; Brunke, B; Hill, A; Griffiths, M W

1998-07-01

Pasteurized and raw milks that had been inoculated at 10(4) cfu/ml with bioluminescent strains of Listeria monocytogenes and Escherichia coli O157:H7 were used in the manufacture of Camembert and Feta cheeses with or without nisin-producing starter culture. Survival of both organisms was determined during the manufacture and storage of Camembert and Feta cheeses at 2 +/- 1 degree C for 65 and 75 d, respectively. Bacterial bioluminescence was used as an indicator to enumerate the colonies plated on selective Listeria agar and on MacConkey agar. Escherichia coli O157:H7 survived the manufacturing process of both cheeses and was present at the end of the storage period in greater numbers than in the initial inoculum. At the end of 75 d of storage, E. coli O157:H7 was found in the brine of Feta cheese. The counts of L. monocytogenes increased as the pH of the Camembert cheese increased, and there were significant differences between the counts from samples taken from the inside and the counts from samples obtained near the surface of the cheese. The Feta cheese that contained nisin was the only cheese in which L. monocytogenes was at the level of the initial inoculum after 75 d of storage.

Botulism challenge studies of a modified atmosphere package for fresh mussels: inoculated pack studies.

PubMed

Newell, C R; Ma, Li; Doyle, Michael

2012-06-01

A series of botulism challenge studies were performed to determine the possibility of production of botulinum toxin in mussels (Mytilus edulis) held under a commercial high-oxygen (60 to 65% O(2)), modified atmosphere packaging (MAP) condition. Spore mixtures of six strains of nonproteolytic Clostridium botulinum were introduced into mussel MAP packages receiving different packaging buffers with or without the addition of lactic acid bacteria. Dye studies and package flipping trials were conducted to ensure internalization of spores by packed mussels. Inoculated mussel packages were stored at normal (4°C) and abusive (12°C) temperatures for 21 and 13 days, respectively, which were beyond the packaged mussels' intended shelf life. Microbiological and chemical analyses were conducted at predetermined intervals (a total of five sampling times at each temperature), including total aerobic plate counts, C. botulinum counts, lactic acid bacterial counts, package headspace gas composition, pH of packaging buffer and mussel meat, and botulinum toxin assays of packaging buffer and mussel meat. Results revealed that C. botulinum inoculated in fresh mussels packed under MAP packaging did not produce toxin, even at an abusive storage temperature and when held beyond their shelf life. No evidence was found that packaging buffers or gas composition influenced the lack of botulinum toxin production in packed mussels.

Chlorhexidine avoids skin bacteria recolonization more than triclosan.

PubMed

Macias, Juan H; Alvarez, Mildred F; Arreguin, Virginia; Muñoz, Juan M; Macias, Alejandro E; Alvarez, Jose A

2016-12-01

We do not know whether differences exist between the residual effect of 2% chlorhexidine in 70% isopropyl alcohol when compared with 1% triclosan in 70% isopropyl alcohol. Using an analytic, longitudinal, controlled, and comparative experimental trial, with blinded measurements, we recruited healthy, adult volunteers from the University of Guanajuato who completed a stabilization phase of skin microbiota and had no history of skin allergies. Four 25-cm 2 areas of the inner surface of the forearms were designated for study: unscrubbed control for establishing baseline bacterial counts, scrubbed control with tridistilled water, scrubbed with chlorhexidine, and scrubbed with triclosan. Quantitative cultures were taken of all the areas at 0, 3, and 24 hours, using agar plates with neutralizing agents. A total of 135 healthy volunteers were tested. At 24 hours, the unscrubbed control counts were 288 CFU/cm 2 , whereas the scrubbed control counts were 96 CFU/cm 2 ; 24 CFU/cm 2 for chlorhexidine and 96 CFU/cm 2 for triclosan (Kruskal-Wallis χ 2 H = 64.27; P <.001). Chlorhexidine is the best antiseptic option when a prolonged antiseptic effect is needed; for instance, when implanting medical devices or performing surgical procedures. Copyright © 2016 Association for Professionals in Infection Control and Epidemiology, Inc. Published by Elsevier Inc. All rights reserved.

Method for Performing Aerobic Plate Counts of Anhydrous Cosmetics Utilizing Tween 60 and Arlacel 80 as Dispersing Agents

PubMed Central

McConville, John F.; Anger, Claude B.; Anderson, David W.

1974-01-01

An aqueous diluent containing Tween 60 and Arlacel 80 gave greater recovery of microorganisms when compared with two common diluents as determined by aerobic plate count of inoculated anhydrous cosmetics. The greater recovery was caused by better dispersion of the anhydrous cosmetics in the diluents. Images PMID:4203790

Microbiology of beef carcasses before and after slaughterline automation.

PubMed Central

Whelehan, O. P.; Hudson, W. R.; Roberts, T. A.

1986-01-01

The bacterial status of beef carcasses at a commercial abattoir was monitored before and after slaughterline automation. Bacterial counts did not differ significantly overall (P greater than 0.05) between the original manual line and the automated line for either morning or afternoon slaughter. On the manual line counts in the morning were lower than those from carcasses slaughtered in the afternoon, but on the automated line there was no difference between morning and afternoon counts. Due to highly significant line X sample site interaction for both morning and afternoon counts, overall differences among sample sites were not found by analysis of variance. However, principal components analysis revealed a significant shift in bacterial contamination among some sites due to slaughterline changes. The incidence of Enterobacteriaceae increased marginally following automation. PMID:3701039

Suitability of optical, physical and chemical measurements for detection of changes in bacterial drinking water quality.

PubMed

Ikonen, Jenni; Pitkänen, Tarja; Miettinen, Ilkka T

2013-10-25

In this study, different optical, physical and chemical measurements were tested for their capacity to detect changes in water quality. The tests included UV-absorbance at 254 nm, absorbance at 420 nm, turbidity, particle counting, temperature, pH, electric conductivity (EC), free chlorine concentration and ATP concentration measurements. Special emphasis was given to investigating the potential for measurement tools to detect changes in bacterial concentrations in drinking water. Bacterial colony counts (CFU) and total bacterial cell counts (TBC) were used as reference methods for assessing the bacterial water quality. The study consists of a series of laboratory scale experiments: monitoring of regrowth of Pseudomonas fluorescens, estimation of the detection limits for optical measurements using Escherichia coli dilutions, verification of the relationships by analysing grab water samples from various distribution systems and utilisation of the measurements in the case of an accidentally contaminated distribution network. We found significant correlations between the tested measurements and the bacterial water quality. As the bacterial contamination of water often co-occurs with the intrusion of matrixes containing mainly non-bacterial components, the tested measurement tools can be considered to have the potential to rapidly detect any major changes in drinking water quality.

Reduction of pasteurization temperature leads to lower bacterial outgrowth in pasteurized fluid milk during refrigerated storage: a case study.

PubMed

Martin, N H; Ranieri, M L; Wiedmann, M; Boor, K J

2012-01-01

Bacterial numbers over refrigerated shelf-life were enumerated in high-temperature, short-time (HTST) commercially pasteurized fluid milk for 15 mo before and 15 mo after reducing pasteurization temperature from 79.4°C (175°F) [corrected] to 76.1°C (169°F). Total bacterial counts were measured in whole fat, 2% fat, and fat-free milk products on the day of processing as well as throughout refrigerated storage (6°C) at 7, 14, and 21 d postprocessing. Mean total bacterial counts were significantly lower immediately after processing as well as at 21 d postprocessing in samples pasteurized at 76.1°C versus samples pasteurized at 79.4°C. In addition to mean total bacterial counts, changes in bacterial numbers over time (i.e., bacterial growth) were analyzed and were lower during refrigerated storage of products pasteurized at the lower temperature. Lowering the pasteurization temperature for unflavored fluid milk processed in a commercial processing facility significantly reduced bacterial growth during refrigerated storage. Copyright © 2012 American Dairy Science Association. Published by Elsevier Inc. All rights reserved.

Suitability of Optical, Physical and Chemical Measurements for Detection of Changes in Bacterial Drinking Water Quality

PubMed Central

Ikonen, Jenni; Pitkänen, Tarja; Miettinen, Ilkka T.

2013-01-01

In this study, different optical, physical and chemical measurements were tested for their capacity to detect changes in water quality. The tests included UV-absorbance at 254 nm, absorbance at 420 nm, turbidity, particle counting, temperature, pH, electric conductivity (EC), free chlorine concentration and ATP concentration measurements. Special emphasis was given to investigating the potential for measurement tools to detect changes in bacterial concentrations in drinking water. Bacterial colony counts (CFU) and total bacterial cell counts (TBC) were used as reference methods for assessing the bacterial water quality. The study consists of a series of laboratory scale experiments: monitoring of regrowth of Pseudomonas fluorescens, estimation of the detection limits for optical measurements using Escherichia coli dilutions, verification of the relationships by analysing grab water samples from various distribution systems and utilisation of the measurements in the case of an accidentally contaminated distribution network. We found significant correlations between the tested measurements and the bacterial water quality. As the bacterial contamination of water often co-occurs with the intrusion of matrixes containing mainly non-bacterial components, the tested measurement tools can be considered to have the potential to rapidly detect any major changes in drinking water quality. PMID:24284353

Dynamic light scattering: A fast and reliable method to analyze bacterial growth during the lag phase.

PubMed

Vargas, Susana; Millán-Chiu, Blanca E; Arvizu-Medrano, Sofía M; Loske, Achim M; Rodríguez, Rogelio

2017-06-01

A comparison between plate counting (PC) and dynamic light scattering (DLS) is reported. PC is the standard technique to determine bacterial population as a function of time; however, this method has drawbacks, such as the cumbersome preparation and handling of samples, as well as the long time required to obtain results. Alternative methods based on optical density are faster, but do not distinguish viable from non-viable cells. These inconveniences are overcome by using DLS. Two different bacteria strains were considered: Escherichia coli and Staphylococcus aureus. DLS was performed at two different illuminating conditions: continuous and intermittent. By the increment of particle size as a function of time, it was possible to observe cell division and the formation of aggregates containing very few bacteria. The scattered intensity profiles showed the lag phase and the transition to the exponential phase of growth, providing a quantity proportional to viable bacteria concentration. The results revealed a clear and linear correlation in both lag and exponential phase, between the Log 10 (colony-forming units/mL) from PC and the Log 10 of the scattered intensity I s from DLS. These correlations provide a good support to use DLS as an alternative technique to determine bacterial population. Copyright © 2017 Elsevier B.V. All rights reserved.

Effects of pipe materials on chlorine-resistant biofilm formation under long-term high chlorine level.

PubMed

Zhu, Zebing; Wu, Chenguang; Zhong, Dan; Yuan, Yixing; Shan, Lili; Zhang, Jie

2014-07-01

Drinking water distribution systems are composed of various pipe materials and may harbor biofilms even in the continuous presence of disinfectants. Biofilms formation on five pipe materials (copper (Cu), polyethylene (PE), stainless steel (STS), cast iron (CI), and concrete-coated polycarbonate (CP)) within drinking water containing 1.20 mg/L free chlorine, was investigated by flow cytometry, heterotrophic plate counts, and denaturing gradient gel electrophoresis analysis. Results showed that the biofilms formation varied in pipe materials. The biofilm formed on CP initially emerged the highest biomass in 12 days, but CI presented the significantly highest biomass after 28 days, and Cu showed the lowest bacterial numbers before 120 days, while STS expressed the lowest bacterial numbers after 159 days. In the biofilm community structure, Moraxella osloensis and Sphingomonas sp. were observed in all the pipe materials while Bacillus sp. was detected except in the CP pipe and Stenotrophomonas maltophila was found from three pipe materials (Cu, PE, and STS). Other bacteria were only found from one or two pipe materials. It is noteworthy that there are 11 opportunistic pathogens in the 17 classified bacterial strains. This research has afforded crucial information regarding the influence of pipe materials on chlorine-resistant biofilm formation.

Relationships between Fungal Biomass and Nitrous Oxide Emission in Upland Rice Soils under No Tillage and Cover Cropping Systems.

PubMed

Zhaorigetu; Komatsuzaki, Masakazu; Sato, Yoshinori; Ohta, Hiroyuki

2008-01-01

The relationships between soil microbial properties and nitrous oxide emission were examined in upland soil under different tillage systems [no tillage (NT), rotary and plow tillage] and cover crop systems (fallow, cereal rye, and hairy vetch) in 2004 and 2005. Microbiological analyses included the determination of soil ergosterol as an indicator of fungal biomass, bacterial plate counting, and MPN estimations of ammonia oxidizers and denitrifiers. The combined practice of NT with rye-cover crop treatment increased fungal biomass but not bacterial populations in 0-10 cm deep soils. Such increase in fungal biomass was not found in 10-20 cm and 20-30 cm deep cover-cropped NT soil. The combined practice of NT with rye-cover cropping resulted in higher in situ N(2)O emission rates compared with rotary- and plow-till treatments. N(2)O flux was positively correlated with soil ergosterol content but not with denitrifier MPN and other soil chemical properties. These results suggested a significant contribution of fungi to N(2)O emission in cover-cropped NT soils.

Human milk antibacterial factors: the effect of temperature on defense systems.

PubMed

Chen, H Y; Allen, J C

2001-01-01

Bovine milk will eventually spoil at refrigeration temperatures, but endogenous or exogenous pathogenic or spoilage bacteria in human milk stored for delayed feeding will die. We investigated the mechanism for these antibacterial properties and their response to high-tempertature, short-time (HTST, 72 degrees C-75 degrees C, 15 sec) and low-temperature long-time (LTLT, 65 degrees C, 30 min) pasteurization. Nonpathogenic Listeria innocua (10(6) cfu/mL) was inoculated into raw and processed bovine and human milk; bacterial plate counts twice weekly determined antibacterial activities. Up to 99% of L. innocua were killed and further growth was inhibited in raw and pasteurized human milk for at least 60 days at 4 degrees C. Reactive IgA antibodies against Listeria antigens were demonstrated by enzyme immunoassay in some human milk samples; sIgA activity against Escherichia coli O antigens was significantly decreased by heat treatments (raw, 1.8; HTST, 1.1; LTLT, 1.3 activity units). Adding human lactoferrin (0.5-20 mg/mL) to the Listeria inoculum (approximately 10(7) cfu/mL) in 1% peptone water did not inhibit bacterial growth.

Effect of cadmium on lake water bacteria as determined by the luciferase assay of adenosine triphosphate

DOE Office of Scientific and Technical Information (OSTI.GOV)

Seyfried, P.L.; Horgan, C.B.L.

1981-10-01

A firefly luciferase assay of bacterial adenosine triphosphate (ATP) was developed to measure the toxic effects of cadmium ions on aquatic organisms. Toxicity was monitored using intracellular (I/C) ATP (in micrograms per litre) as well as plate counts (colony-forming units per millilitre). The bacteria, which belonged mainly to the families Enterobacteriaceae and Pseudomonadaceae, exhibited varying degrees of resistance to up to 100 ppm cadmium when grown in a glucose-salts medium at pH 6.8. Among the organisms tested, cadmium resistance decreased in the following order: Pseudomonas vesicularis > P. aeruginosa > Enterobacter sp. > P. fluorescens > Chromobacter sp. > Serratiamore » sp. A rise in the pH of the growth medium from 5 to 7 resulted in increased toxicity of cadmium.« less

Microbial changes in patients with acute periodontal abscess after treatment detected by PadoTest.

PubMed

Eguchi, T; Koshy, G; Umeda, M; Iwanami, T; Suga, J; Nomura, Y; Kawanami, M; Ishikawa, I

2008-03-01

To investigate changes in bacterial counts in subgingival plaque from patients with acute periodontal abscess by IAI-PadoTest. Ninety-one patients were randomly allocated to either test or control groups. In all the patients, pockets with acute periodontal abscess were irrigated with sterilized physiological saline, and in the test group, 2% minocycline hydrochloride ointment was applied once into the pocket in addition. Subgingival plaque samples were collected by paper point before treatment and 7 days after treatment. The total bacterial count was determined and Actinobacillus actinomycetemcomitans, Porphyromonas gingivalis, Treponema denticola, and Tannerella forsythia, were detected using IAI-PadoTest, a DNA/RNA probe method. The total bacterial count decreased in both groups, with a significant decrease in the test group. The counts and number of sites positive for P. gingivalis, T. forsythia and T. denticola significantly decreased in the test group after treatment, compared with those in the control group. Pocket depth decreased in the both groups, with a statistically significant decrease in the test group. Topical treatment with minocycline in pockets with acute periodontal abscess was effective in reducing the bacterial counts as shown by the microbiological investigation using PadoTest 4.5.

Evaluation of a clay-based acidic bedding conditioner for dairy cattle bedding.

PubMed

Proietto, R L; Hinckley, L S; Fox, L K; Andrew, S M

2013-02-01

This study investigated the effects of a clay-based acidic bedding conditioner on sawdust bedding pH, dry matter (DM), environmental pathogen counts, and environmental bacterial counts on teat ends of lactating dairy cows. Sixteen lactating Holstein cows were paired based on parity, days in milk, milk yield, and milk somatic cell count, and were negative for the presence of an intramammary pathogen. Within each pair, cows were randomly assigned to 1 of 2 treatments with 3-wk periods in a crossover design. Treatment groups consisted of 9 freestalls per group bedded with either untreated sawdust or sawdust with a clay-based acidic bedding conditioner, added at 3- to 4-d intervals over each 21-d period. Bedding and teat ends were aseptically sampled on d 0, 1, 2, 7, 14, and 21 for determination of environmental bacterial counts. At the same time points, bedding was sampled for DM and pH determination. The bacteria identified in the bedding material were total gram-negative bacteria, Streptococcus spp., and coliform bacteria. The bacteria identified on the teat ends were Streptococcus spp., coliform bacteria, and Klebsiella spp. Teat end score, milk somatic cell count, and intramammary pathogen presence were measured weekly. Bedding and teat cleanliness, environmental high and low temperatures, and dew point data were collected daily. The bedding conditioner reduced the pH, but not the DM, of the sawdust bedding compared with untreated sawdust. Overall environmental bacterial counts in bedding were lower for treated sawdust. Total bacterial counts in bedding and on teat ends increased with time over both periods. Compared with untreated sawdust, the treated bedding had lower counts of total gram-negative bacteria and streptococci, but not coliform counts. Teat end bacterial counts were lower for cows bedded on treated sawdust for streptococci, coliforms, and Klebsiella spp. compared with cows bedded on untreated sawdust. The clay-based acidic bedding conditioner reduced environmental pathogens in sawdust bedding and teat ends without affecting teat end integrity. Copyright © 2013 American Dairy Science Association. Published by Elsevier Inc. All rights reserved.

Slit lamps and lenses: a potential source of nosocomial infections?

PubMed

Sobolewska, Bianka; Buhl, Michael; Liese, Jan; Ziemssen, Focke

2018-01-30

The aim of the study was to evaluate the bacterial contamination level of contact surfaces on slit lamps and the grip areas of lenses. Within unannounced audits, two regions of the slit lamps (headrest and joystick), indirect ophthalmoscopy devices, and ultrasound probes were obtained with rayon-tipped swab. Non-contact lenses used for indirect fundoscopy were pressed on RODAC (Replicate Organism Detection and Counting) plates. One hundred and eighty-one surfaces were sampled. The total number of colony-forming units was assessed and bacterial species were identified. Spa-typing and antimicrobial susceptibility testing were performed from Staphylococcus aureus isolates. Among the total bacterial isolates from ophthalmological equipment (lenses: 51 of 78, slit lamps: 43 of 88, ophthalmoscopy helmets: 3 of 8, ultrasound probes: 2 of 7), coagulase-negative staphylococci (CNS) was most frequently found, followed by Micrococcus spp. (lenses vs. slit lamps: P < 0.001 and P = 0.01, respectively). The bacterial contamination of lenses (76%) was significantly higher than that of slit lamps (54%) (P < 0.003). A significantly higher contamination with CNS was observed on lenses from residents vs. from consultants (78% vs. 35%, P = 0.01). A total of seven different spa-types of S. aureus were isolated. No correlation was found between S. aureus contamination of different ophthalmological equipments (Spearman's rank correlation coefficient, ρ = 0.04, P = 0.75). Methicillin-resistant S. aureus was not detected. Bacterial species of the normal skin flora were isolated from the ophthalmological equipment. The bacterial contamination of the portable devices was significantly higher than that of slit lamps. Therefore, proper hygiene of the mobile instruments should be monitored in order to prevent transmission of bacteria in residents and consultants.

IN THAILAND.

PubMed

Luksamijarulkul, Pipat; Suknongbung, Siranee; Vatanasomboon, Pisit; Sujirarut, Dusit

2017-03-01

A large number of migrants have move to cities in Thailand seeking employment. These people may be at increased risk for environmental health problems. We studied the health status, environmental living conditions and microbial indoor air quality (IAQ) among selected groups of migrant workers and their households in Mueang District, Samut Sakhon, central Thailand. We conducted a cross sectional study of 240 migrant workers and their households randomly selected by multistage sampling. The person responsible for hygiene at each studied household was interviewed using a structured questionnaire. Two indoor air samples were taken from each household (480 indoor air samples) to determine bacterial and fungal counts using a Millipore air tester; 240 outdoor air samples were collected for comparison. Ninety-nine point six percent of study subjects were Myanmar, 74.2% were aged 21-40 years, 91.7% had a primary school level education or lower and 53.7% had stayed in Thailand less than 5 years. Eight point three percent had a history of an underlying disease, 20.8% had a recent history of pulmonary tuberculosis in a family member within the previous year. Forty-three point eight percent had a current illness related to IAQ during a previous month. Twenty-one point three were current cigarette smokers, 15.0% were current alcohol consumers, and 5.0% exercises ≥3 times per week. Forty-nine point two percent never opened the windows of their bedrooms or living rooms for ventilation, 45% never cleaned their window screens, and 38.3% never put their pillows or mattresses in the sunlight. The mean(±SD) air bacterial count was 230(±229) CFU/m3 (outdoor air = 128±82 CFU/ m3), and the mean fungal count was 630(±842) CFU/m3 (outdoor air = 138±94 CFU/ m3). When the bacterial and fungal counts were compared with the guidelines of the American Conference of Governmental Industrial Hygienists, the bacterial counts in 6.5% of houses surveyed and the fungal counts in 28.8% of house surveyed were higher than the recommended levels (<500 CFU/m3). Bacterial and fungal counts in the sample households were not significantly correlated with household hygiene practice scores (p>0.05). There was a positive correlation between bacterial counts and fungal counts in household air samples, r=0.28, p<0.001.

Gastrointestinal health effects associated with the consumption of drinking water produced by point-of-use domestic reverse-osmosis filtration units.

PubMed Central

Payment, P; Franco, E; Richardson, L; Siemiatycki, J

1991-01-01

During a prospective epidemiological study of gastrointestinal health effects associated with the consumption of drinking water produced by reverse-osmosis domestic units, a correlation was demonstrated between the bacterial counts on R2A medium incubated at 35 degrees C and the reported gastrointestinal symptoms in families who used these units. A univariate correlation was found with bacterial counts on R2A medium at 20 degrees C but was confounded by the bacterial counts at 35 degrees C. Other variables, such as family size and amount of water consumed, were not independently explanatory of the rate of illness. These observations raise concerns for the possibility of increased disease associated with certain point-of-use treatment devices for domestic use when high levels of bacterial growth occur. PMID:2059052

Molecular identification and quantification of bacteria from endodontic infections using real-time polymerase chain reaction.

PubMed

Blome, B; Braun, A; Sobarzo, V; Jepsen, S

2008-10-01

It was the aim of the present study to evaluate root canal samples for the presence and numbers of specific species as well as for total bacterial load in teeth with chronic apical periodontitis using quantitative real-time polymerase chain reaction (PCR). Forty adult patients with one radiographically documented periapical lesion were included. Twenty teeth presented with primary infections and 20 with secondary infections, requiring retreatment. After removal of necrotic pulp tissue or root canal filling, a first bacterial sample was obtained. Following chemo-mechanical root canal preparation a second sample was taken and a third sample was obtained after 14 days of intracanal dressing with calcium hydroxide. Analysis by real-time PCR enabled the quantification of total bacterial counts and of nine selected species. Root canals with primary infections harbored significantly more bacteria (by total bacterial count) than teeth with secondary infections (P < 0.05). Mean total bacterial count in the retreatment group was 2.1 x 10(6) and was significantly reduced following root canal preparation (3.6 x 10(4)) and intracanal dressing (1.4 x 10(5)). Corresponding values for primary infections were: 4.6 x 10(7), 3.6 x 10(4), and 6.9 x 10(4). The numbers of the selected bacteria and their detection frequency were also significantly reduced. Root canals with primary infections contained a higher bacterial load. Chemo-mechanical root canal preparation reduced bacterial counts by at least 95%.

Defective plastic infection-control barriers and faulty technique may cause PSP plate contamination used in digital intraoral radiography.

PubMed

Kuperstein, Arthur S

2012-09-01

Fifty-two disinfected photostimulable phosphor (PSP) plates in plastic barrier envelopes were evaluated for contamination following placement in 30 study participants. Forty-four plates were acceptable for use in the study. The risk factor was the abundant oropharyngeal microbial flora and its ability to breach infection-control barrier sheaths. The presence of bacterial colonies on an agar plate was used to determine bacterial contamination and the presence of any growth indicated failure of the barrier envelope. Before clinical placement of the plates, quality review of the PSP plates revealed defects in the integrity of 4 barrier envelopes most likely caused by forceps-related damage or failure to achieve a uniform seal during manufacturing. These defects allowed substantial contamination. Contamination also occurred as a result of failure to extract the PSP plate from the barrier envelope cleanly. Of the 44 barriers with no obvious signs of a defect, 3 produced bacterial growth following culture. The authors concluded that digital sensor sheathed in barrier envelopes remain a potential source of contamination. PSP plates must be disinfected between removal from a contaminated barrier envelope (used in a patient) and placement in a new barrier envelope. In addition, placement into the barrier envelope should ideally be carried out under aseptic conditions. Finally, the integrity of each sealed barrier envelope must be verified visually. Copyright © 2012. Published by Mosby, Inc. All rights reserved.

RAPA: a novel in vitro method to evaluate anti-bacterial skin cleansing products.

PubMed

Ansari, S A; Gafur, R B; Jones, K; Espada, L A; Polefka, T G

2010-04-01

Development of efficacious anti-bacterial skin cleansing products has been limited by the availability of a pre-clinical (in vitro) method to predict clinical efficacy adequately. We report a simple and rapid method, designated as rapid agar plate assay (RAPA), that uses the bacteriological agar surface as a surrogate substrate for skin and combines elements of two widely used in vivo (clinical) methods (Agar Patch and Cup Scrub). To simulate the washing of the human hand or forearm skin with the test product, trypticase soy agar plates were directly washed with the test product and rinsed under running tap water. After air-drying the washed plates, test bacteria (Staphylococcus aureus or Escherichia coli) were applied and the plates were incubated at 37 degrees C for 18-24 h. Using S. aureus as the test organism, anti-bacterial bar soap containing triclocarbanilide showed a strong linear relationship (R(2) = 0.97) between bacterial dose and their per cent reduction. A similar dose-response relationship (R(2) = 0.96) was observed for anti-bacterial liquid hand soap against E. coli. RAPA was able to distinguish between anti-bacterial products based on the nature and level of actives in them. In limited comparative tests, results obtained by RAPA were comparable with the results obtained by clinical agar patch and clinical cup scrub methods. In conclusion, RAPA provides a simple, rugged and reproducible in vitro method for testing the relative efficacy of anti-bacterial skin cleansing products with a likelihood of comparable clinical efficacy. Further testing is warranted to improve the clinical predictability of this method.

Radiolabel ratio method for measuring pulmonary clearance of intratracheal bacterial challenges

DOE Office of Scientific and Technical Information (OSTI.GOV)

LaForce, F.M.; Boose, D.S.

Calculation of bacterial clearance is a fundamental step in any study of in situ lung antibacterial defenses. A method is described whereby about 85% of a radiolabeled bacterial inoculum was consistently introduced into the bronchopulmonary tree of a mouse by the intratracheal route. Mice were then killed 1 and 4 hours later; their lungs were removed aseptically and homogenized, and viable bacteria and radiolabel counts were determined. Radiolabel counts fell slowly, and more than 80% of the original radiolabel was still present in homogenized lung samples from animals sacrificed 4 hours after challenge. Bacteria/isotope ratios for the bacterial inoculum andmore » homogenized lung samples from animals sacrificed immediately after challenge were very similar. Bacterial clearance values were the same whether computed from bacterial counts alone or according to a radiolabel ratio method whereby the change in the bacteria/isotope ratio in ground lung aliquots was divided by a similar ratio from bacteria used to inoculate animals. Some contamination resulted from oral streptococci being swept into the bronchopulmonary free during the aspiration process. This contamination was not a problem when penicillin was incorporated into the agar and penicillin-resistant strains were used for the bacterial challenges.« less

SURVIVAL OF SALMONELLA SPECIES IN RIVER WATER.

EPA Science Inventory

The survival of four Salmonella strains in river water microcosms was monitored using culturing techniques, direct counts, whole cell hybridization, scanning electron microscopy, and resuscitation techniques via the direct viable count method and flow cytrometry. Plate counts of...

Immunological techniques as tools to characterize the subsurface microbial community at a trichloroethylene contaminated site

DOE Office of Scientific and Technical Information (OSTI.GOV)

Fliermans, C.B.; Dougherty, J.M.; Franck, M.M.

Effective in situ bioremediation strategies require an understanding of the effects pollutants and remediation techniques have on subsurface microbial communities. Therefore, detailed characterization of a site`s microbial communities is important. Subsurface sediment borings and water samples were collected from a trichloroethylene (TCE) contaminated site, before and after horizontal well in situ air stripping and bioventing, as well as during methane injection for stimulation of methane-utilizing microorganisms. Subsamples were processed for heterotrophic plate counts, acridine orange direct counts (AODC), community diversity, direct fluorescent antibodies (DFA) enumeration for several nitrogen-transforming bacteria, and Biolog {reg_sign} evaluation of enzyme activity in collected water samples.more » Plate counts were higher in near-surface depths than in the vadose zone sediment samples. During the in situ air stripping and bioventing, counts increased at or near the saturated zone, remained elevated throughout the aquifer, but did not change significantly after the air stripping. Sporadic increases in plate counts at different depths as well as increased diversity appeared to be linked to differing lithologies. AODCs were orders of magnitude higher than plate counts and remained relatively constant with depth except for slight increases near the surface depths and the capillary fringe. Nitrogen-transforming bacteria, as measured by serospecific DFA, were greatly affected both by the in situ air stripping and the methane injection. Biolog{reg_sign} activity appeared to increase with subsurface stimulation both by air and methane. The complexity of subsurface systems makes the use of selective monitoring tools imperative.« less

Method of detecting and counting bacteria

NASA Technical Reports Server (NTRS)

Picciolo, G. L.; Chappelle, E. W. (Inventor)

1976-01-01

An improved method is provided for determining bacterial levels, especially in samples of aqueous physiological fluids. The method depends on the quantitative determination of bacterial adenosine triphosphate (ATP) in the presence of nonbacterial ATP. The bacterial ATP is released by cell rupture and is measured by an enzymatic bioluminescent assay. A concentration technique is included to make the method more sensitive. It is particularly useful where the fluid to be measured contains an unknown or low bacteria count.

Microbiological Surveillance of Operation Theatres: Five Year Retrospective Analysis from a Tertiary Care Hospital in North India

PubMed Central

Najotra, Dipender Kaur; Malhotra, Aneeta Singh; Slathia, Poonam; Raina, Shivani; Dhar, Ashok

2017-01-01

Introduction: Microbiological contamination of air and environment in the operation theaters (OTs) are major risk factor for surgical site and other hospital-associated infections. Objectives: The aim was to identify bacterial colonization of surfaces and equipment and to determine the microbial contamination of air in the OTs of a tertiary care hospital. Materials and Methods: Five years (January 2010–December 2014) retrospective analysis of the data obtained from routine microbiological surveillance of the five OTs of the hospital was done. Surface samples were taken with wet swabs from different sites and equipment. Bacterial species were isolated and identified by conventional methods. Air quality surveillance of OTs was done by settle plate method. Results: A total of 4387 samples were collected from surfaces and articles of various OTs. Out of these only 195 (4.4%), samples showed bacterial growth and yielded 210 isolates. The predominant species isolated was Bacillus with 184 (87.6%) isolates followed by coagulase-negative Staphylococcus 17 (8.1%), Staphylococcus aureus 6 (2.9%), and Enteroccoccus spp. 3 (1.4%). Analysis of the OT air samples showed least colony forming unit (cfu) rate of air (27 cfu/m3) in ophthalmology OT and highest rate of 133 cfu/m3 in general surgery OT. Conclusion: The study shows that OTs of our hospital showed a very low bacterial contamination rate on surface swabbing and a cfu count per m3 of air well within permissible limits. PMID:28904915

Live/Dead Bacterial Spore Assay Using DPA-Triggered Tb Luminescence

NASA Technical Reports Server (NTRS)

Ponce, Adrian

2003-01-01

A method of measuring the fraction of bacterial spores in a sample that remain viable exploits DPA-triggered luminescence of Tb(3+) and is based partly on the same principles as those described earlier. Unlike prior methods for performing such live/dead assays of bacterial spores, this method does not involve counting colonies formed by cultivation (which can take days), or counting of spores under a microscope, and works whether or not bacterial spores are attached to other small particles (i.e., dust), and can be implemented on a time scale of about 20 minutes.

Introduction of sample tubes with sodium azide as a preservative for ethyl glucuronide in urine.

PubMed

Luginbühl, Marc; Weinmann, Wolfgang; Al-Ahmad, Ali

2017-09-01

Ethyl glucuronide (EtG) is a direct alcohol marker, which is widely used for clinical and forensic applications, mainly for abstinence control. However, the instability of EtG in urine against bacterial degradation or the post-collectional synthesis of EtG in contaminated samples may cause false interpretation of EtG results in urine samples. This study evaluates the potential of sodium azide in tubes used for urine collection to hinder degradation of ethyl glucuronide by bacterial metabolism taking place during growth of bacterial colonies. The tubes are part of a commercial oral fluid collection device. The sampling system was tested with different gram-positive and gram-negative bacterial species previously observed in urinary tract infections, such as Escherichia coli, Staphylococcus aureus, Enterecoccus faecalis, Staphylococcus epidermidis, Klebsiella pneumoniae, Enterobacter cloacae, and Pseudomonas aeruginosa. Inhibition of bacterial growth by sodium azide, resulting in lower numbers of colony forming units compared to control samples, was observed for all tested bacterial species. To test the prevention of EtG degradation by the predominant pathogen in urinary tract infection, sterile-filtered urine and deficient medium were spiked with EtG, and inoculated with E. coli prior to incubation for 4 days at 37 °C in tubes with and without sodium azide. Samples were collected every 24 hours, during four consecutive days, whereby the colony forming units (CFU) were counted on Columbia blood agar plates, and EtG was analyzed by LC-MS/MS. As expected, EtG degradation was observed when standard polypropylene tubes were used for the storage of contaminated samples. However, urine specimens collected in sodium azide tubes showed no or very limited bacterial growth and no EtG degradation. As a conclusion, sodium azide is useful to reduce bacterial growth of gram-negative and gram-positive bacteria. It inhibits the degradation of EtG by E. coli and can be used for the stabilization of EtG in urine samples.

SURVIVAL OF SALMONELLA SPECIES IN RIVER WATER

EPA Science Inventory

The survival of four Salmonella strains in river water microcosms was monitored by culturing techniques, direct counts, whole-cell hybridization, scanning electron microscopy, and resuscitation techniques via the direct viable count method and flow cytometry. Plate counts of bact...

Water Sources and Their Protection from the Impact of Microbial Contamination in Rural Areas of Beijing, China

PubMed Central

Ye, Bixiong; Yang, Linsheng; Li, Yonghua; Wang, Wuyi; Li, Hairong

2013-01-01

Bacterial contamination of drinking water is a major public health problem in rural China. To explore bacterial contamination in rural areas of Beijing and identify possible causes of bacteria in drinking water samples, water samples were collected from wells in ten rural districts of Beijing, China. Total bacterial count, total coliforms and Escherichia coli in drinking water were then determined and water source and wellhead protection were investigated. The bacterial contamination in drinking water was serious in areas north of Beijing, with the total bacterial count, total coliforms and Escherichia coli in some water samples reaching 88,000 CFU/mL, 1,600 MPN/100 mL and 1,600 MPN/100 mL, respectively. Water source types, well depth, whether the well was adequately sealed and housed, and whether wellhead is above or below ground were the main factors influencing bacterial contamination levels in drinking water. The bacterial contamination was serious in the water of shallow wells and wells that were not closed, had no well housing or had a wellhead below ground level. The contamination sources around wells, including village dry toilets and livestock farms, were well correlated with bacterial contamination. Total bacterial counts were affected by proximity to sewage ditches and polluting industries, however, proximity to landfills did not influence the microbial indicators. PMID:23462436

Possible association of Bifidobacterium and Lactobacillus in the gut microbiota of patients with major depressive disorder.

PubMed

Aizawa, Emiko; Tsuji, Hirokazu; Asahara, Takashi; Takahashi, Takuya; Teraishi, Toshiya; Yoshida, Sumiko; Ota, Miho; Koga, Norie; Hattori, Kotaro; Kunugi, Hiroshi

2016-09-15

Bifidobacterium and Lactobacillus in the gut have been suggested to have a beneficial effect on stress response and depressive disorder. We examined whether these bacterial counts are reduced in patients with major depressive disorder (MDD) than in healthy controls. Bifidobacterium and Lactobacillus counts in fecal samples were estimated in 43 patients and 57 controls using bacterial rRNA-targeted reverse transcription-quantitative polymerase chain reaction The patients had significantly lower Bifidobacterium counts (P=0.012) and tended to have lower Lactobacillus counts (P=0.067) than the controls. Individuals whose bacterial counts below the optimal cut-off point (9.53 and 6.49log10 cells/g for Bifidobacterium and Lactobacillus, respectively) were significantly more common in the patients than in the controls for both bacteria (Bifidobacterium: odds ratio 3.23, 95% confidence interval [CI] 1.38-7.54, P=0.010; Lactobacillus: 2.57, 95% CI 1.14-5.78, P=0.027). Using the same cut-off points, we observed an association between the bacterial counts and Irritable bowel syndrome. Frequency of fermented milk consumption was associated with higher Bifidobacterium counts in the patients. The findings should be interpreted with caution since effects of gender and diet were not fully taken into account in the analysis. Our results provide direct evidence, for the first time, that individuals with lower Bifidobacterium and/or Lactobacillus counts are more common in patients with MDD compared to controls. Our findings provide new insight into the pathophysiology of MDD and will enhance future research on the use of pro- and prebiotics in the treatment of MDD. Copyright © 2016 Elsevier B.V. All rights reserved.

The role of bacteria in lactational mastitis and some considerations of the use of antibiotic treatment.

PubMed

Kvist, Linda J; Larsson, Bodil Wilde; Hall-Lord, Marie Louise; Steen, Anita; Schalén, Claes

2008-04-07

The role of bacterial pathogens in lactational mastitis remains unclear. The objective of this study was to compare bacterial species in breast milk of women with mastitis and of healthy breast milk donors and to evaluate the use of antibiotic therapy, the symptoms of mastitis, number of health care contacts, occurrence of breast abscess, damaged nipples and recurrent symptoms in relation to bacterial counts. In this descriptive study, breast milk from 192 women with mastitis (referred to as cases) and 466 breast milk donors (referred to as controls) was examined bacteriologically and compared using analytical statistics. Statistical analyses were also carried out to test for relationships between bacteriological content and clinical symptoms as measured on scales, prescription of antibiotics, the number of care contacts, occurrence of breast abscess and recurring symptoms. Five main bacterial species were found in both cases and controls: coagulase negative staphylococci (CNS), viridans streptococci, Staphylococcus aureus (S. aureus), Group B streptococci (GBS) and Enterococcus faecalis. More women with mastitis had S. aureus and GBS in their breast milk than those without symptoms, although 31% of healthy women harboured S. aureus and 10% had GBS. There were no significant correlations between bacterial counts and the symptoms of mastitis as measured on scales. There were no differences in bacterial counts between those prescribed and not prescribed antibiotics or those with and without breast abscess. GBS in breast milk was associated with increased health care contacts (p = 0.02). Women with >/= 10(7) cfu/L CNS or viridans streptococci in their breast milk had increased odds for damaged nipples (p = 0.003). Many healthy breastfeeding women have potentially pathogenic bacteria in their breast milk. Increasing bacterial counts did not affect the clinical manifestation of mastitis; thus bacterial counts in breast milk may be of limited value in the decision to treat with antibiotics as results from bacterial culture of breast milk may be difficult to interpret. These results suggest that the division of mastitis into infective or non-infective forms may not be practically feasible. Daily follow-up to measure the subsidence of symptoms can help detect those in need of antibiotics.

Exploring the potential environmental functions of viable but non-culturable bacteria.

PubMed

Su, Xiaomei; Chen, Xi; Hu, Jinxing; Shen, Chaofeng; Ding, Linxian

2013-12-01

A conventional plate count is the most commonly employed method to estimate the number of living bacteria in environmental samples. In fact, judging the level of viable culture by plate count is limited, because it is often several orders of magnitude less than the number of living bacteria actually present. Most of the bacteria are in "viable but non-culturable" (VBNC) state, whose cells are intact and alive and can resuscitate when surrounding conditions are more favorable. The most exciting recent development in resuscitating VBNC bacteria is a bacterial cytokine, namely, the resuscitation-promoting factor (Rpf), secreted by Micrococcus luteus, which promotes the resuscitation and growth of high G+C Gram-positive organisms, including some species of the genus Mycobacterium. However, most of studies deal with VBNC bacteria only from the point of view of medicine and epidemiology. It is therefore of great significance to research whether these VBNC state bacteria also possess some useful environmental capabilities, such as degradation, flocculation, etc. Further studies are needed to elucidate the possible environmental role of the VBNC bacteria, rather than only considering their role as potential pathogens from the point view of epidemiology and public health. We have studied the resuscitation of these VBNC bacteria in polluted environments by adding culture supernatant containing Rpf from M. luteus, and it was found that, as a huge microbial resource, VBNC bacteria could provide important answers to dealing with existing problems of environmental pollution. This mini-review will provide new insight for considering the potentially environmental functions of VBNC bacteria.

Individual based simulations of bacterial growth on agar plates

NASA Astrophysics Data System (ADS)

Ginovart, M.; López, D.; Valls, J.; Silbert, M.

2002-03-01

The individual based simulator, INDividual DIScrete SIMulations (INDISIM) has been used to study the behaviour of the growth of bacterial colonies on a finite dish. The simulations reproduce the qualitative trends of pattern formation that appear during the growth of Bacillus subtilis on an agar plate under different initial conditions of nutrient peptone concentration, the amount of agar on the plate, and the temperature. The simulations are carried out by imposing closed boundary conditions on a square lattice divided into square spatial cells. The simulator studies the temporal evolution of the bacterial population possible by setting rules of behaviour for each bacterium, such as its uptake, metabolism and reproduction, as well as rules for the medium in which the bacterial cells grow, such as concentration of nutrient particles and their diffusion. The determining factors that characterize the structure of the bacterial colony patterns in the presents simulations, are the initial concentrations of nutrient particles, that mimic the amount of peptone in the experiments, and the set of values for the microscopic diffusion parameter related, in the experiments, to the amount of the agar medium.

Usefulness of Cellular Analysis of Bronchoalveolar Lavage Fluid for Predicting the Etiology of Pneumonia in Critically Ill Patients

PubMed Central

Hong, Hyo-Lim; Kim, Sung-Han; Huh, Jin Won; Sung, Heungsup; Lee, Sang-Oh; Kim, Mi-Na; Jeong, Jin-Yong; Lim, Chae-Man; Kim, Yang Soo; Woo, Jun Hee; Koh, Younsuck

2014-01-01

Background The usefulness of bronchoalveolar lavage (BAL) fluid cellular analysis in pneumonia has not been adequately evaluated. This study investigated the ability of cellular analysis of BAL fluid to differentially diagnose bacterial pneumonia from viral pneumonia in adult patients who are admitted to intensive care unit. Methods BAL fluid cellular analysis was evaluated in 47 adult patients who underwent bronchoscopic BAL following less than 24 hours of antimicrobial agent exposure. The abilities of BAL fluid total white blood cell (WBC) counts and differential cell counts to differentiate between bacterial and viral pneumonia were evaluated using receiver operating characteristic (ROC) curve analysis. Results Bacterial pneumonia (n = 24) and viral pneumonia (n = 23) were frequently associated with neutrophilic pleocytosis in BAL fluid. BAL fluid median total WBC count (2,815/µL vs. 300/µL, P<0.001) and percentage of neutrophils (80.5% vs. 54.0%, P = 0.02) were significantly higher in the bacterial pneumonia group than in the viral pneumonia group. In ROC curve analysis, BAL fluid total WBC count showed the best discrimination, with an area under the curve of 0.855 (95% CI, 0.750–0.960). BAL fluid total WBC count ≥510/µL had a sensitivity of 83.3%, specificity of 78.3%, positive likelihood ratio (PLR) of 3.83, and negative likelihood ratio (NLR) of 0.21. When analyzed in combination with serum procalcitonin or C-reactive protein, sensitivity was 95.8%, specificity was 95.7%, PLR was 8.63, and NLR was 0.07. BAL fluid total WBC count ≥510/µL was an independent predictor of bacterial pneumonia with an adjusted odds ratio of 13.5 in multiple logistic regression analysis. Conclusions Cellular analysis of BAL fluid can aid early differential diagnosis of bacterial pneumonia from viral pneumonia in critically ill patients. PMID:24824328

RT-PCR quantification of periodontal pathogens in crack users and non-users.

PubMed

Casarin, M; Antoniazzi, R P; Vaucher, R A; Feldens, C A; Zanatta, F B

2017-04-01

To compare counts of Aggregatibacter actinomycetemcomitans, Prevotella intermedia, Porphyromonas gingivalis and Fusobacterium nucleatum between crack users and non-users. A cross-sectional study was conducted involving seventy-four crack cocaine users and eighty-one non-users matched for age, gender and tobacco use. Demographic and clinical variables were analysed. Subgingival bacterial samples were collected from four sites with the greatest probing depths and were analysed using real-time polymerase chain reaction. No significant difference was found in the prevalence of total counts for each bacterial species analysed between groups. However, crack users had a 1.85 (95% CI: 1.03-3.31), 2.19 (95% CI 1.24-3.88), 2.53 (95% CI 1.27-5.04) and 2.40 (95% CI 1.22-4.75) greater probability of having the higher counts (≥75th percentile) for Aggregatibacter actinomycetemcomitans, Porphyromonas gingivalis, Prevotella intermedia and Fusobacterium nucleatum, respectively. Although some crack users had higher (>75th percentile) bacterial counts for Aggregatibacter actinomycetemcomitans, Porphyromonas gingivalis, Prevotella intermedia and Fusobacterium nucleatum, total counts did not differ between crack users and non-users, leading to the hypothesis that the higher occurrence of periodontitis on crack users may be related to other non-bacterial factors. © 2016 John Wiley & Sons A/S. Published by John Wiley & Sons Ltd.

Simultaneous construction of PCR-DGGE-based predictive models of Listeria monocytogenes and Vibrio parahaemolyticus on cooked shrimps.

PubMed

Liao, C; Peng, Z Y; Li, J B; Cui, X W; Zhang, Z H; Malakar, P K; Zhang, W J; Pan, Y J; Zhao, Y

2015-03-01

The aim of this study was to simultaneously construct PCR-DGGE-based predictive models of Listeria monocytogenes and Vibrio parahaemolyticus on cooked shrimps at 4 and 10°C. Calibration curves were established to correlate peak density of DGGE bands with microbial counts. Microbial counts derived from PCR-DGGE and plate methods were fitted by Baranyi model to obtain molecular and traditional predictive models. For L. monocytogenes, growing at 4 and 10°C, molecular predictive models were constructed. It showed good evaluations of correlation coefficients (R(2) > 0.92), bias factors (Bf ) and accuracy factors (Af ) (1.0 ≤ Bf ≤ Af ≤ 1.1). Moreover, no significant difference was found between molecular and traditional predictive models when analysed on lag phase (λ), maximum growth rate (μmax ) and growth data (P > 0.05). But for V. parahaemolyticus, inactivated at 4 and 10°C, molecular models show significant difference when compared with traditional models. Taken together, these results suggest that PCR-DGGE based on DNA can be used to construct growth models, but it is inappropriate for inactivation models yet. This is the first report of developing PCR-DGGE to simultaneously construct multiple molecular models. It has been known for a long time that microbial predictive models based on traditional plate methods are time-consuming and labour-intensive. Denaturing gradient gel electrophoresis (DGGE) has been widely used as a semiquantitative method to describe complex microbial community. In our study, we developed DGGE to quantify bacterial counts and simultaneously established two molecular predictive models to describe the growth and survival of two bacteria (Listeria monocytogenes and Vibrio parahaemolyticus) at 4 and 10°C. We demonstrated that PCR-DGGE could be used to construct growth models. This work provides a new approach to construct molecular predictive models and thereby facilitates predictive microbiology and QMRA (Quantitative Microbial Risk Assessment). © 2014 The Society for Applied Microbiology.

Survival of B. Horneckiae Spores Under Ground-simulated Space Conditions

NASA Technical Reports Server (NTRS)

Schanche, Bradley

2012-01-01

To prevent forward contamination and maintain the scientific integrity of future life detection missions, it is important to characterize and attempt to eliminate terrestrial microorganisms associated with exploratory spacecraft and landing vehicles. Among the organisms isolated from spacecraft-associated habitats, spore-forming microbes are highly resistant to various physical and chemical conditions, which include ionizing and UV radiation, desiccation and oxidative stress, and the harsh environment of outer space or planetary surfaces. Recently a radiation resistant, spore forming bacterial isolate, Bacillus horneckiae, was isolated from a clean room of the Kennedy Space Center where the Phoenix spacecraft was assembled. The exceptionally high tolerance of extreme conditions demonstrated by sporeforming bacteria highlighted the need to assess the viability of these microbes in situ (in real) space. The proposed BOSS (Biofilm Organisms Surfing Space) project aims to understand the mechanisms by which biofilm forming organisms, such as B. horneckiae, will potentially be able to withstand harsh space conditions. As previously stated, the spore producing ability of these species gives them increased survivability to harsh conditions. Some of the spores will have the protective exosporium layer artificially removed before the test to determine if the existence of this layer significantly changes the survivability during the mission. In preparation for that experiment, we analyzed spores which were exposed during a ground simulation, the EXPOSE R2 Biofilm Organisms Surfing Space (BOSS). Previous to exposure, spores were deposited onto spacecraft grade aluminum coupons in a spore suspension calculated to contain between 10(exp 7) and 10(exp 8) spores. This precursor series will be used to establish a baseline survivability function for comparison with the future flight tests during EXPOSE-R. For each coupon, a 10% polyvinyl alcohol (PVA) film was applied and peeled from the coupon to recover the spores. One hundred µl of sterile 10% PVA was applied to the surface of the coupon and allowed to dry for 1 hour at 37 C. The films were then removed using sterile scalpel and forceps and placed into a glass test tube containing 2 milliliters of sterile deionized water. The PVA film process was then repeated on each coupon one additional time to ensure recovery of the majority of spores. The second PVA film was added in the same glass tube as in the previous round. If the spores remained 100% viable, the test tubes should now contain between 5 X 10(exp 6) and 5 X 10(exp 7) spores per millimeter; however, it is expected that some loss of viability has occurred. In order to assess this loss, the number of colony forming, viable spores was counted. To count the colony forming units (CFUs), the spore containing solution was diluted in a process of 10-fold serial dilution by mixing successive solutions in a 100 microliter spore suspension to 900 microliter deionized H2O ratio. A sample dilution series revealed that 10(exp -3) and 10(exp -4) concentrations would be necessary for an accurate CFU count to be taken. For those two concentrations, a spread on a TSA plate was prepared and incubated at 32 C. For the samples exposed to UV radiation, the cell survivability was too low to establish a count from 100 microliter spread plating. Instead, no dilutions were performed and the entire 2 milliliter spore suspension was plated and incubated at 32 C. The plate's CFU counts were taken at 24 hours and 48 hours from the time of plating. At the end of the CFU counting the total surviving spores in each sample were calculated based on the number of CFUs that were observed per 100 microliters, or per 2 milliliters for the UV irradiated samples. The results of these calculations are shown in Figures 1 and 2.

Microbial diversity and prevalence of foodborne pathogens in cheap and junk foods consumed by primary schoolchildren.

PubMed

Kim, M J; Kim, S A; Kang, Y S; Hwang, I G; Rhee, M S

2013-07-01

Aerobic plate counts (APC), coliforms, Bacillus cereus, Escherichia coli and eight foodborne pathogens were tested in 1008 cheap and junk foods, including candies, dried cakes, chewing gum, chocolate, dried and seasoned seafood, ice cream, and sugary foods. APCs were positive for 342 samples (33·9%), and the majority of the counts were 2-3 log CFU g(-1) or ml(-1) (average: 1·10 log CFU g(-1) or ml(-1) ). Most samples (97·3%) contained no coliforms (average: 0·07 log CFU g(-1) or ml(-1) ). Bacillus cereus was detected in 68 samples (average: 0·14 log CFU g(-1) or ml(-1) ). Escherichia coli and Listeria monocytogenes were detected in 6 and 1 samples, respectively, whereas other foodborne pathogens were not isolated. The highest bacterial counts were associated with dried and seasoned seafood products and dried cakes, suggesting that appropriate regulations of these food types should be considered. Cheap and junk foods were produced mainly in developing countries, but there were no significant differences in the bacterial counts among different countries of origin. The presence of foodborne pathogens may pose a risk for children. These results suggest that there is cause for deeper concern about the safety of these foods and that effective countermeasures should be established to improve their microbiological safety. Food safety is especially important for children, but only limited information is available about the microbiological quality of cheap and junk foods that are consumed frequently by primary schoolchildren (e.g. dried cakes, candies and chocolates). The present study investigated the microbial quality of cheap and junk foods, and our results indicate that these foods are a potential health risk for children, therefore, deeper concern about the safety of these foods and effective countermeasures should be established to improve their microbiological safety. The present study may contribute to the development of an appropriate child food safety management system. © 2013 The Society for Applied Microbiology.

Short communication: Bacterial counts in recycled manure solids bedding replaced daily or deep packed in freestalls.

PubMed

Sorter, D E; Kester, H J; Hogan, J S

2014-05-01

An experiment was conducted to compare bacterial counts of mastitis pathogens in deep-packed manure solids bedding with those in manure solids bedding replaced daily from mattresses. Eighteen Holstein cows were housed in 1 pen with 18 stalls. One row of 9 stalls was equipped with mattresses topped with bedding. The back one-third of these stalls toward the alleyway was covered in 25 mm of recycled manure solids, which was removed daily for the next 6 d and replaced with bedding from the brisket board and lunge space areas of stalls. The second row of 9 stalls was bedded for 3 wk with 100 to 150 mm of deep-pack recycled manure bedding from which only fecal matter was removed daily. After 3 wk, bedding treatments were changed between rows in a switchback design. Mean total gram-negative bacterial counts did not differ between treatments throughout the experiment. Coliform and Klebsiella spp. bacterial counts were lower in daily replaced bedding compared with deep pack across the experiment and on each of d 0, 1, 2, and 6. Streptococcal counts were reduced in daily replacement stalls compared with deep-pack stalls on d 0 and greater in daily replacement stalls compared with deep-pack stalls on d 1, 2, and 6. Daily replacement of recycled manure bedding from the back one-third of the stalls appeared to be an effective approach to reducing exposure to coliforms, specifically Klebsiella, but not streptococci. However, bacterial counts in bedding from both treatments were elevated throughout the trial and resulted in considerable risk for exposure to teats and development of intramammary infections. Copyright © 2014 American Dairy Science Association. Published by Elsevier Inc. All rights reserved.

Analysis of the bacterial strains using Biolog plates in the contaminated soil from Riyadh community.

PubMed

Al-Dhabaan, Fahad Abdullah M; Bakhali, Ali Hassan

2017-05-01

Routine manufacture, detonation and disposal of explosives in land and groundwater have resulted in complete pollution. Explosives are xenobiotic compounds, being toxic to biological systems, and their recalcitrance leads to persistence in the environment. The methods currently used for the remediation of explosive contaminated sites are expensive and can result in the formation of toxic products. The present study aimed to investigate the bacterial strains using the Biolog plates in the soil from the Riyadh community. The microbial strains were isolated using the spread plate technique and were identified using the Biolog method. In this study we have analyzed from bacterial families of soil samples, obtained from the different sites in 5 regions at Explosive Institute. Our results conclude that Biolog MicroPlates were developed for the rapid identification of bacterial isolates by sole-carbon source utilization and can be used for the identification of bacteria. Out of five communities, only four families of bacteria indicate that the microbial community lacks significant diversity in region one from the Riyadh community in Saudi Arabia. More studies are needed to be carried out in different regions to validate our results.

Anatomical characteristics of teats and premilking bacterial counts of teat skin swabs of primiparous cows exposed to different types of bedding.

PubMed

Guarín, J F; Baumberger, C; Ruegg, P L

2017-02-01

Bacterial populations of teat skin are associated with risk of intramammary infection and may be influenced by anatomical characteristics of teats. The objective of this study was to evaluate associations of selected anatomical characteristics of teats with bacterial counts of teat skin of cows exposed to different types of bedding. Primarily primiparous Holstein cows (n = 128) were randomly allocated to 4 pens within a single barn. Each pen contained 1 type of bedding [new sand (NES), recycled sand (RS), deep-bedded manure solids (DBMS), and shallow-bedded manure solids over foam core mattresses (SBMS)]. During a single farm visit udders (n = 112) were scored for hygiene and 1 front (n = 112) and 1 rear teat (n = 111) of each enrolled cow were scored for hyperkeratosis (HK). Teat length, teat barrel diameter, and teat apex diameter were measured and teat skin swabs were systematically collected for microbiological analysis. Linear type evaluation data for udders of each cow were retrieved for each cow. Teat position (front or rear) was associated with occurrence of clinical mastitis during the 12 mo before the farm visit and more cases occurred in front quarters. The proportion of udders that were classified as clean (score 1 or 2) was 68, 82, 54, and 95% for cows housed in pens containing NES, RS, SBMS, and DBMS, respectively. No association was found between HK score and teat position and no association was found between HK score and teat skin bacterial count. Bacterial counts of teat skin swabs from front teats of cows in pens containing RS and SBMS were significantly less than those of rear teats of cows in pens containing DBMS or NES. Teat skin bacterial counts were significantly greater for swabs obtained from teats of cows with udder hygiene scores of 3 and 4 as compared with swabs obtained from cows with cleaner udders. Of all udder conformation traits evaluated, only narrower rear teat placement was positively associated with bacterial counts on teat skin. Copyright © 2017 American Dairy Science Association. Published by Elsevier Inc. All rights reserved.

[Analysis on 2011 quality control results on aerobic plate count of microbiology laboratories in China].

PubMed

Han, Haihong; Li, Ning; Li, Yepeng; Fu, Ping; Yu, Dongmin; Li Zhigang; Du, Chunming; Guo, Yunchang

2015-01-01

To test the aerobic plate count examining capability of microbiology laboratories, to ensure the accuracy and comparability of quantitative bacteria examination results, and to improve the quality of monitoring. The 4 different concentration aerobic plate count piece samples were prepared and noted as I, II, III and IV. After homogeneity and stability tests, the samples were delivered to monitoring institutions. The results of I, II, III samples were logarithmic transformed, and evaluated with Z-score method using the robust average and standard deviation. The results of IV samples were evaluated as "satisfactory" when reported as < 10 CFU/piece or as "not satisfactory" otherwise. Pearson χ2 test was used to analyze the ratio results. 309 monitoring institutions, which was 99.04% of the total number, reported their results. 271 institutions reported a satisfactory result, and the satisfactory rate was 87.70%. There was no statistical difference in satisfactory rates of I, II and III samples which were 81.52%, 88.30% and 91.40% respectively. The satisfactory rate of IV samples was 93.33%. There was no statistical difference in satisfactory rates between provincial and municipal CDC. The quality control program has provided scientific data that the aerobic plate count capability of the laboratories meets the requirements of monitoring tasks.

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.

Short communication: Environmental mastitis pathogen counts in freestalls bedded with composted and fresh recycled manure solids.

PubMed

Cole, K J; Hogan, J S

2016-02-01

An experiment was conducted to compare bacterial counts of environmental mastitis pathogens in composted recycled manure solids bedding with those in fresh recycled manure solids. Eighteen Holstein cows were housed in 1 pen with 18 stalls. One row of 9 freestalls included mattresses and was bedded weekly with composted recycled manure solids. The second row of 9 freestalls included mattresses and was bedded weekly with fresh recycled manure solids. The back one-third of stalls toward the alleyway was covered in 25 to 50 mm of bedding. Samples were taken from the back one-third of 4 stalls for both treatments on d 0, 1, 2, and 6 of each week. After 3 wk, bedding treatments were switched between rows, making the total duration 6 wk. Mean total gram-negative bacterial counts were approximately 0.5 log10 cfu/g of dry matter lower in the composted recycled manure solids on d 0 compared with fresh recycled manure solids. Klebsiella species, coliform, and Streptococcus species counts were at least 1.0 log10 cfu/g of dry matter lower in composted compared with fresh recycled manure solids on d 0. Only gram-negative bacterial counts on d 1 were reduced in composted recycled manure solids compared with fresh recycled manure solids. Differences were not observed between treatments in gram-negative bacterial, coliform, Klebsiella species, or Streptococcus species counts on d 2 and 6. Ash content was higher in composted recycled manure solids compared with fresh recycled manure solids on d 0, 1, 2, and 6. Despite the increase in ash after composting, bacterial counts of mastitis pathogens in composted recycled manure solids were comparable with those in fresh recycled manure when used as freestall bedding. Copyright © 2016 American Dairy Science Association. Published by Elsevier Inc. All rights reserved.

Mobile laminar air flow screen for additional operating room ventilation: reduction of intraoperative bacterial contamination during total knee arthroplasty.

PubMed

Sossai, D; Dagnino, G; Sanguineti, F; Franchin, F

2011-12-01

Surgical site infections are important complications in orthopedic surgery. A mobile laminar air flow (LAF) screen could represent a useful addition to an operating room (OR) with conventional turbulent air ventilation (12.5 air changes/h), as it could decrease the bacterial count near the operating field. The purpose of this study was to evaluate LAF efficacy at reducing bacterial contamination in the surgical area during 34 total knee arthroplasties (TKAs). The additional unit was used in 17 operations; the LAF was positioned beside the operating table between two of the surgeons, with the air flow directed towards the surgical area (wound). The whole team wore conventional OR clothing and the correct hygiene procedures and rituals were used. Bacterial air contamination (CFU/m(3)) was evaluated in the wound area in 17 operations with the LAF unit and 17 without the LAF unit. The LAF unit reduced the mean bacterial count in the wound area from 23.5 CFU/m(3) without the LAF to 3.5 CFU/m(3) with the LAF (P < 0.0001), which is below the suggested limit for an OR with ultraclean laminar ventilation. There were no significant differences in the mean bacterial count in the instrument table area: 28.6 CFU/m(3) were recorded with the LAF (N = 6) unit and 30.8 CFU/m(3) (N = 6) without the LAF unit (P = 0.631). During six operations with LAF and six without LAF, particle counts were performed and the number of 0.5 μm particles was analyzed. The particle counts decreased significantly when the LAF unit was used (P = 0.003). When a mobile LAF unit was added to the standard OR ventilation, bacterial contamination of the wound area significantly decreased to below the accepted level for an ultraclean OR, preventing SSI infections.

Inactivation of Escherichia coli and coliform bacteria in traditional brass and earthernware water storage vessels.

PubMed

Tandon, Puja; Chhibber, Sanjay; Reed, Robert H

2005-07-01

The detection and enumeration of indicator bacteria such as Escherichia coli is used to assess the extent of faecal contamination of drinking water. On the basis of this approach, the effectiveness of storing water contaminated with faecal indicator bacteria in brass or earthern vessels (mutkas) of the type used in rural India have been investigated. Suspensions of bacteria in sterile distilled water were maintained for up to 48 h in each vessel and enumerated by surface plate counts on nutrient agar (non-selective) and several selective coliform media at 37 degrees C either under standard aerobic conditions, or under conditions designed to neutralise reactive oxygen species (ROS), e.g. using an anaerobic cabinet to prepare plates of pre-reduced growth medium or by inclusion of sodium pyruvate in the growth medium, with incubation of aerobically-prepared plates in an anaerobic jar. The counts obtained for E. coli decreased on short-term storage in a brass mutka; counts for selective media were lower than for equivalent counts for non-selective medium, with ROS-neutralised conditions giving consistently higher counts than aerobic incubation. However, after 48 h, no bacteria were cultivable under any conditions. Similar results were obtained using water from environmental sources in the Panjab, and from rural households where brass and earthern mutkas are used for storage of drinking water, with enumeration on selective coliform media (presumptive total coliforms). In all cases results indicated that, while storage of water in a brass mutka can inactivate E. coli and coliforms over a 48 h period, standard aerobic plate counting using selective media may not be fully effective in enumerating sub-lethally damaged bacteria.

Effect of photoactivated disinfection with a light-emitting diode on bacterial species and biofilms associated with periodontitis and peri-implantitis.

PubMed

Eick, Sigrun; Markauskaite, Giedre; Nietzsche, Sandor; Laugisch, Oliver; Salvi, Giovanni E; Sculean, Anton

2013-05-01

To determine the effect of photoactivated disinfection (PAD) using toluidine blue and a light-emitting diode (LED) in the red spectrum (wave length at 625-635 nm) on species associated with periodontitis and peri-implantitis and bacteria within a periodontopathic biofilm. Sixteen single microbial species including 2 Porphyromonas gingivalis and 2 Aggregatibacter actinomycetemcomitans and a multispecies mixture consisting of 12 species suspended in saline without and with 25% human serum were exposed to PAD. Moreover, single-species biofilms consisting of 2 P. gingivalis and 2 A. actinomycetemcomitans strains and a multi-species biofilm on 24-well-plates, grown on titanium discs and in artificial periodontal pockets were exposed to PAD with and without pretreatment with 0.25% hydrogen peroxide. Changes in the viability were determined by counting the colony forming units (cfu). PAD reduced the cfu counts in saline by 1.42 log₁₀ after LED application for 30s and by 1.99 log₁₀ after LED application for 60s compared with negative controls (each p<0.001). Serum did not inhibit the efficacy of PAD. PAD reduced statistically significantly (p<0.05) the cfu counts of the P. gingivalis biofilms. The viability of the A. actinomycetemcomitans biofilms and the multi-species biofilms was statistically significantly decreased when PAD was applied after a pretreatment with 0.25% hydrogen peroxide. The biofilm formed in artificial pockets was more sensitive to PAD with and without pretreatment with hydrogen peroxide compared with those formed on titanium discs. PAD using a LED was effective against periodontopathic bacterial species and reduced viability in biofilms but was not able to completely destroy complex biofilms. The use of PAD following pretreatment with hydrogen peroxide resulted in an additional increase in the antimicrobial activity which may represent a new alternative to treat periodontal and peri-implant infections thus warranting further testing in clinical studies. Copyright © 2012 Elsevier B.V. All rights reserved.

Applications of chemiluminescence to bacterial analysis

NASA Technical Reports Server (NTRS)

Searle, N. D.

1975-01-01

Luminol chemiluminescence method for detecting bacteria was based on microbial activation of the oxidation of the luminol monoanion by hydrogen peroxide. Elimination of the prior lysing step, previously used in the chemiluminescence technique, was shown to improve considerably the reproducibility and accuracy of the method in addition to simplifying it. An inexpensive, portable photomultiplier detector was used to measure the maximum light intensity produced when the sample is added to the reagent. Studies of cooling tower water show that the luminol chemiluminescence technique can be used to monitor changes in viable cell population both under normal conditions and during chlorine treatment. Good correlation between chemiluminescence and plate counts was also obtained in the analysis of process water used in paper mills. This method showed good potential for monitoring the viable bacteria populations in activated sludge used in waste treatment plants to digest organic matter.

Rapid depletion of dissolved oxygen in 96-well microtiter plate Staphylococcus epidermidis biofilm assays promotes biofilm development and is influenced by inoculum cell concentration.

PubMed

Cotter, John J; O'Gara, James P; Casey, Eoin

2009-08-01

Biofilm-related research using 96-well microtiter plates involves static incubation of plates indiscriminate of environmental conditions, making oxygen availability an important variable which has not been considered to date. By directly measuring dissolved oxygen concentration over time we report here that dissolved oxygen is rapidly consumed in Staphylococcus epidermidis biofilm cultures grown in 96-well plates irrespective of the oxygen concentration in the gaseous environment in which the plates are incubated. These data indicate that depletion of dissolved oxygen during growth of bacterial biofilm cultures in 96-well plates may significantly influence biofilm production. Furthermore higher inoculum cell concentrations are associated with more rapid consumption of dissolved oxygen and higher levels of S. epidermidis biofilm production. Our data reveal that oxygen depletion during bacterial growth in 96-well plates may significantly influence biofilm production and should be considered in the interpretation of experimental data using this biofilm model.

A non-invasive in vivo imaging system to study dissemination of bioluminescent Yersinia pestis CO92 in a mouse model of pneumonic plague

PubMed Central

Sha, Jian; Rosenzweig, Jason A.; Kirtley, Michelle L.; van Lier, Christina J.; Fitts, Eric C.; Kozlova, Elena V.; Erova, Tatiana E.; Tiner, Bethany L.; Chopra, Ashok K.

2012-01-01

The gold standard in microbiology for monitoring bacterial dissemination in infected animals has always been viable plate counts. This method, despite being quantitative, requires sacrificing the infected animals. Recently, however, an alternative method of in vivo imaging of bioluminescent bacteria (IVIBB) for monitoring microbial dissemination within the host has been employed. Yersina pestis is a Gram-negative bacterium capable of causing bubonic, septicemic, and pneumonic plague. In this study, we compared the conventional counting of bacterial colony forming units (cfu) in the various infected tissues to IVIBB in monitoring Y. pestis dissemination in a mouse model of pneumonic plague. By using a transposon mutagenesis system harboring the luciferase (luc) gene, we screened approximately 4000 clones and obtained a fully virulent, luc-positive Y. pestis CO92 (Y. pestis-luc2) reporter strain in which transposition occurred within the largest pMT1 plasmid which possesses murine toxin and capsular antigen encoding genes. The aforementioned reporter strain and the wild-type CO92 exhibited similar growth curves, formed capsule based on immunofluorescence microscopy and flow cytometry, and had a similar LD50. Intranasal infection of mice with 15 LD50 of CO92-luc2 resulted in animal mortality by 72 h, and an increasing number of bioluminescent bacteria were observed in various mouse organs over a 24–72 h period when whole animals were imaged. However, following levofloxacin treatment (10 mg/kg/day) for 6 days 24 h post infection, no luminescence was observed after 72 h of infection, indicating that the tested antimicrobial killed bacteria preventing their detection in host peripheral tissues. Overall, we demonstrated that IVIBB is an effective and non-invasive way of monitoring bacterial dissemination in animals following pneumonic plague having strong correlation with cfu, and our reporter CO92-luc2 strain can be employed as a useful tool to monitor the efficacy of antimicrobial countermeasures in real time. PMID:23063826

Bile Salt-induced Biofilm Formation in Enteric Pathogens: Techniques for Identification and Quantification.

PubMed

Nickerson, Kourtney P; Faherty, Christina S

2018-05-06

Biofilm formation is a dynamic, multistage process that occurs in bacteria under harsh environmental conditions or times of stress. For enteric pathogens, a significant stress response is induced during gastrointestinal transit and upon bile exposure, a normal component of human digestion. To overcome the bactericidal effects of bile, many enteric pathogens form a biofilm hypothesized to permit survival when transiting through the small intestine. Here we present methodologies to define biofilm formation through solid-phase adherence assays as well as extracellular polymeric substance (EPS) matrix detection and visualization. Furthermore, biofilm dispersion assessment is presented to mimic the analysis of events triggering release of bacteria during the infection process. Crystal violet staining is used to detect adherent bacteria in a high-throughput 96-well plate adherence assay. EPS production assessment is determined by two assays, namely microscopy staining of the EPS matrix and semi-quantitative analysis with a fluorescently-conjugated polysaccharide binding lectin. Finally, biofilm dispersion is measured through colony counts and plating. Positive data from multiple assays support the characterization of biofilms and can be utilized to identify bile salt-induced biofilm formation in other bacterial strains.

Bacterial and chemical oxidation of pyritic mine tailings at low temperatures

NASA Astrophysics Data System (ADS)

Elberling, Bo; Schippers, Axel; Sand, Wolfgang

2000-02-01

Microbial and chemical sulfide oxidation activity and oxygen consumption was investigated in the active layer of pyritic mine tailings at Nanisivik Mine, located in a permafrost area on Baffin Island in northern Canada. Samples of tailings were collected up to a depth of 60 cm in mid-August 1998 at 4 sites, for which the metabolic activity of sulfur- and iron-oxidizing leaching bacteria besides the chemical pyrite oxidation activity were measured on 39 tailings samples and 7 samples from a natural pyritic site by calorimetry. The tailings of varying age and water content were deposited under alkaline conditions. In situ oxygen uptake rates were measured at the tailings surface every third day, prior to sampling. In addition, cell counts of iron(II), sulfur, and thiosulfate oxidizing, lithotrophic bacteria and chemoorganotrophic microorganisms were determined quantitatively by the most-probable-number technique or by agar-plating. Results show consistent pyrite oxidation rates based on in situ oxygen uptake rates, and laboratory heat output measurements. Litho- and organotrophic bacteria were found in the tailings. Calorimetric measurements revealed that the present bacterial activity is responsible for approximately one third of the ongoing oxidation. Although leaching bacteria have previously been found in the Arctic, this study is the first to prove the significance of bacterial activity in the overall pollution resulting from tailings deposited in the Arctic.

Antibacterial and Antiadhesive Activities of Extracts from Edible Plants against Soft Drink Spoilage by Asaia spp.

PubMed

Antolak, Hubert; Czyzowska, Agata; Kregiel, Dorota

2017-01-01

This study was conducted to investigate the antibacterial and antiadhesive activities of ethanol extracts from five edible plant parts: cinnamon bark ( Cinnamomum zeylanicum ), licorice root ( Glycyrrhiza radix ), nettle leaves ( Urtica dioica ), green tea leaves ( Camellia sinensis ), and elderberry flowers ( Sambucus nigra ). The chemical constituents of the extracts were identified using high-performance liquid chromatography and liquid chromatography plus mass spectrometry. Six strains of Asaia lannensis and Asaia bogorensis bacteria isolated from spoiled commercial fruit-flavored noncarbonated mineral water were used. Bacterial adhesion to polystyrene as an attachment substrate in culture media supplemented with 10% plant extract was evaluated using luminometric measurement of the ATP extracted from adhered cells. The viability of the adhered and planktonic cells was assessed using the plate count method, and the relative adhesion coefficient was calculated. All tested crude extracts contained flavonols (kaempferol, quercetin, and their derivatives), flavanols (catechin and derivatives), flavanones (glabrol, licorice glycoside A, and liquiritin), and phenolic acids (gallic, quinic, chlorogenic, neochlorogenic, caffeic, coumaric, and ferulic). The culture medium with 10% elderberry extract provided the least favorable environment for all tested bacterial strains. Extracts from green tea, cinnamon, and licorice also had significant inhibitory effects on the adhesion of the tested bacterial strains. This research suggests that the addition of selected edible plant extracts could improve the microbial stability of noncarbonated soft drinks.

Evaluation of the viability of Lactobacillus spp. after the production of different solid dosage forms.

PubMed

Brachkova, Mariya I; Duarte, Aida; Pinto, João F

2009-09-01

The work aims to provide evidence on the viability of Lactobacillus spp. and a spore form of Bacillus subtilis from nonprocessed bacteria to coated dosage forms (i.e., mini-tablets, pellets, and their coated forms). Lactobacillus spp. were cultivated overnight in MRS broth (10(9) cfu/mL) and B. subtilis spores were produced on plate count agar (10(7) cfu/mL) for 2 weeks. Bacteria and spores were freeze-dried in skim milk enriched with glycerol. The cakes were further processed into tablets (2.5 mm diameter) by direct compression with or without microcrystalline cellulose and inulin. Pellets (1-1.4 mm diameter) were produced by extrusion-spheronization of bacterial and spore suspensions with microcrystalline cellulose, lactose, inulin, and skim milk. Both tablets and pellets were film coated. The properties of the dosage forms, particularly the bacterial viability, were evaluated immediately after production and throughout storage for 6 months at 4 degrees C. The study has shown that for an adequate stabilization of the bacteria a protective matrix (e.g., skim milk) and cryoprotectors (e.g., glycerol) must be present at early stages of bacterial de-hydration. Tabletting had a less deleterious effect (<2 log units) on bacteria when compared to pelletization (in some cases 3 log units). Enteric coating (15%, w/w) of either tablets or pellets did not affect the viability of the bacteria.

Analysis of Bacterial Community Structure in Sulfurous-Oil-Containing Soils and Detection of Species Carrying Dibenzothiophene Desulfurization (dsz) Genes

PubMed Central

Duarte, Gabriela Frois; Rosado, Alexandre Soares; Seldin, Lucy; de Araujo, Welington; van Elsas, Jan Dirk

2001-01-01

The selective effects of sulfur-containing hydrocarbons, with respect to changes in bacterial community structure and selection of desulfurizing organisms and genes, were studied in soil. Samples taken from a polluted field soil (A) along a concentration gradient of sulfurous oil and from soil microcosms treated with dibenzothiophene (DBT)-containing petroleum (FSL soil) were analyzed. Analyses included plate counts of total bacteria and of DBT utilizers, molecular community profiling via soil DNA-based PCR-denaturing gradient gel electrophoresis (PCR-DGGE), and detection of genes that encode enzymes involved in the desulfurization of hydrocarbons, i.e., dszA, dszB, and dszC.Data obtained from the A soil showed no discriminating effects of oil levels on the culturable bacterial numbers on either medium used. Generally, counts of DBT degraders were 10- to 100-fold lower than the total culturable counts. However, PCR-DGGE showed that the numbers of bands detected in the molecular community profiles decreased with increasing oil content of the soil. Analysis of the sequences of three prominent bands of the profiles generated with the highly polluted soil samples suggested that the underlying organisms were related to Actinomyces sp., Arthrobacter sp., and a bacterium of uncertain affiliation. dszA, dszB, and dszC genes were present in all A soil samples, whereas a range of unpolluted soils gave negative results in this analysis. Results from the study of FSL soil revealed minor effects of the petroleum-DBT treatment on culturable bacterial numbers and clear effects on the DBT-utilizing communities. The molecular community profiles were largely stable over time in the untreated soil, whereas they showed a progressive change over time following treatment with DBT-containing petroleum. Direct PCR assessment revealed the presence of dszB-related signals in the untreated FSL soil and the apparent selection of dszA- and dszC-related sequences by the petroleum-DBT treatment. PCR-DGGE applied to sequential enrichment cultures in DBT-containing sulfur-free basal salts medium prepared from the A and treated FSL soils revealed the selection of up to 10 distinct bands. Sequencing a subset of these bands provided evidence for the presence of organisms related to Pseudomonas putida, a Pseudomonas sp., Stenotrophomonas maltophilia, and Rhodococcus erythropolis. Several of 52 colonies obtained from the A and FSL soils on agar plates with DBT as the sole sulfur source produced bands that matched the migration of bands selected in the enrichment cultures. Evidence for the presence of dszB in 12 strains was obtained, whereas dszA and dszC genes were found in only 7 and 6 strains, respectively. Most of the strains carrying dszA or dszC were classified as R. erythropolis related, and all revealed the capacity to desulfurize DBT. A comparison of 37 dszA sequences, obtained via PCR from the A and FSL soils, from enrichments of these soils, and from isolates, revealed the great similarity of all sequences to the canonical (R. erythropolis strain IGTS8) dszA sequence and a large degree of internal conservation. The 37 sequences recovered were grouped in three clusters. One group, consisting of 30 sequences, was minimally 98% related to the IGTS8 sequence, a second group of 2 sequences was slightly different, and a third group of 5 sequences was 95% similar. The first two groups contained sequences obtained from both soil types and enrichment cultures (including isolates), but the last consisted of sequences obtained directly from the polluted A soil. PMID:11229891

Effect of surgical hand scrub time on subsequent bacterial growth.

PubMed

Wheelock, S M; Lookinland, S

1997-06-01

In this experimental study, the researchers evaluated the effect of surgical hand scrub time on subsequent bacterial growth and assessed the effectiveness of the glove juice technique in a clinical setting. In a randomized crossover design, 25 perioperative staff members scrubbed for two or three minutes in the first trial and vice versa in the second trial, after which the wore sterile surgical gloves for one hour under clinical conditions. The researchers then sampled the subjects' nondominant hands for bacterial growth, cultured aliquots from the sampling solution, and counted microorganisms. Scrubbing for three minutes produced lower mean log bacterial counts than scrubbing for two minutes. Although the mean bacterial count differed significantly (P = .02) between the two-minute and three-minute surgical hand scrub times, it fell below 0.5 log, which is the threshold for practical and clinical significance. This finding suggests that a two-minute surgical hand scrub is clinically as effective as a three-minute surgical had scrub. The glove juice technique demonstrated sensitivity and reliability in enumerating bacteria on the hands of perioperative staff members in a clinical setting.

Project environmental microbiology as related to planetary quarantine

NASA Technical Reports Server (NTRS)

Pflug, I. J.

1974-01-01

Microbiological analyses of soil particles allow for the following conclusions: (1) there is a considerable range in the values of aerobic, mesophilic microbial counts associated with different size soil fractions; (2) as soil particle size increases, there is an increase in the mean microbial concentration per particle; (3) plate counts of aerobic, mesophilic organisms in unheated soils yielded a mean concentration of about six organisms per particle for the smallest soil fraction; (4) aerobic, mesophilic counts for sonicated particles heated at 80 C for 20 minutes yielded mean values of about two organisms per particle for the smallest particles; (5) some actinomycetes associated with the soil fractions could survive dry heat treatment at 110 C for one hour; and (6) soil particles stored under ambient laboratory conditions for 2.5 years aerobic, mesophilic plate counts which were comparable or slightly greater than the counts for more recently collected soil.

A colorimetric assay of 1-aminocyclopropane-1-carboxylate (ACC) based on ninhydrin reaction for rapid screening of bacteria containing ACC deaminase.

PubMed

Li, Z; Chang, S; Lin, L; Li, Y; An, Q

2011-08-01

1-Aminocyclopropane-1-carboxylate (ACC) deaminase activity is an efficient marker for bacteria to promote plant growth by lowering ethylene levels in plants. We aim to develop a method for rapidly screening bacteria containing ACC deaminase, based on a colorimetric ninhydrin assay of ACC. A reliable colorimetric ninhydrin assay was developed to quantify ACC using heat-resistant polypropylene chimney-top 96-well PCR plates, having the wells evenly heated in boiling water, preventing accidental contamination from boiling water and limiting evaporation. With this method to measure bacterial consumption of ACC, 44 ACC-utilizing bacterial isolates were rapidly screened out from 311 bacterial isolates that were able to grow on minimal media containing ACC as the sole nitrogen source. The 44 ACC-utilizing bacterial isolates showed ACC deaminase activities and belonged to the genus Burkholderia, Pseudomonas or Herbaspirillum. Determination of bacterial ACC consumption by the PCR-plate ninhydrin-ACC assay is a rapid and efficient method for screening bacteria containing ACC deaminase from a large number of bacterial isolates. The PCR-plate ninhydrin-ACC assay extends the utility of the ninhydrin reaction and enables a rapid screening of bacteria containing ACC deaminase from large numbers of bacterial isolates. © 2011 The Authors. Letters in Applied Microbiology © 2011 The Society for Applied Microbiology.

Designing primers and evaluation of the efficiency of propidium monoazide - Quantitative polymerase chain reaction for counting the viable cells of Lactobacillus gasseri and Lactobacillus salivarius.

PubMed

Lai, Chieh-Hsien; Wu, Sih-Rong; Pang, Jen-Chieh; Ramireddy, Latha; Chiang, Yu-Cheng; Lin, Chien-Ku; Tsen, Hau-Yang

2017-07-01

The purpose of this study is to evaluate the efficiency of using propidium monoazide (PMA) real-time quantitative polymerase chain reaction (qPCR) to count the viable cells of Lactobacillus gasseri and Lactobacillus salivarius in probiotic products. Based on the internal transcription spacer and 23S rRNA genes, two primer sets specific for these two Lactobacillus species were designed. For a probiotic product, the total deMan Rogosa Sharpe plate count was 8.65±0.69 log CFU/g, while for qPCR, the cell counts of L. gasseri and L. salivarius were 8.39±0.14 log CFU/g and 8.57±0.24 log CFU/g, respectively. Under the same conditions, for its heat-killed product, qPCR counts for L. gasseri and L. salivarius were 6.70±0.16 log cells/g and 7.67±0.20 log cells/g, while PMA-qPCR counts were 5.33±0.18 log cells/g and 5.05±0.23 log cells/g, respectively. For cell dilutions with a viable cell count of 8.5 log CFU/mL for L. gasseri and L. salivarius, after heat killing, the PMA-qPCR count for both Lactobacillus species was near 5.5 log cells/mL. When the PMA-qPCR counts of these cell dilutions were compared before and after heat killing, although some DNA might be lost during the heat killing, significant qPCR signals from dead cells, i.e., about 4-5 log cells/mL, could not be reduced by PMA treatment. Increasing PMA concentrations from 100 μM to 200 μM or light exposure time from 5 minutes to 15 minutes had no or, if any, only minor effect on the reduction of qPCR signals from their dead cells. Thus, to differentiate viable lactic acid bacterial cells from dead cells using the PMA-qPCR method, the efficiency of PMA to reduce the qPCR signals from dead cells should be notable. Copyright © 2016. Published by Elsevier B.V.

Microbial community dynamics during fermentation of doenjang-meju, traditional Korean fermented soybean.

PubMed

Jung, Ji Young; Lee, Se Hee; Jeon, Che Ok

2014-08-18

Bacterial and fungal community dynamics, along with viable plate counts and water content, were investigated in the exterior and interior regions of doenjang-meju, traditional Korean fermented soybean, during its fermentation process. Measurement of viable cells showed that the meju molding equipment might be an important source of bacterial cells (mostly Bacillus) during doenjang-meju fermentation, whereas fungi might be mostly derived from the fermentation environment including incubation shelves, air, and rice straws. Community analysis using rRNA-targeted pyrosequencing revealed that Bacillus among bacteria and Mucor among fungi were predominant in both the exterior and interior regions of doenjang-meju during the early fermentation period. Bacteria such as Ignatzschineria, Myroides, Enterococcus, Corynebacterium, and Clostridium and fungi such as Geotrichum, Scopulariopsis, Monascus, Fusarium, and eventually Aspergillus were mainly detected as the fermentation progressed. Bacillus, an aerobic bacterial group, was predominant in the exterior regions during the entire fermentation period, while anaerobic, facultative anaerobic, and microaerobic bacteria including Enterococcus, Lactobacillus, Clostridium, Myroides, and Ignatzschineria were much more abundant in the interior regions. Principal component analysis (PCA) also indicated that the bacterial communities in the exterior and interior regions were clearly differentiated, suggesting that aeration might be an important factor in determining the bacterial communities during doenjang-meju fermentation. However, PCA showed that fungal communities were not separated in the exterior and interior regions and Pearson's correlation coefficients showed that the major fungal taxa had significantly positive (Mucor and Geotrichum) or negative (Aspergillus) correlations with the water content during doenjang-meju fermentation, indicating that water content might be a significant factor in determining the fungal communities during doenjang-meju fermentation. Copyright © 2014 Elsevier B.V. All rights reserved.

Bacteria permeabilization and disruption caused by sludge reduction technologies evaluated by flow cytometry.

PubMed

Foladori, P; Tamburini, S; Bruni, L

2010-09-01

Technologies proposed in the last decades for the reduction of the sludge production in wastewater treatment plants and based on the mechanism of cell lysis-cryptic growth (physical, mechanical, thermal, chemical, oxidative treatments) have been widely investigated at lab-, pilot- and, in some cases, at full-scale but the effects on cellular lysis have not always been demonstrated in depth. The research presented in this paper aims to investigate how these sludge reduction technologies affect the integrity and permeabilization of bacterial cells in sludge using flow cytometry (FCM), which permits the rapid and statistically accurate quantification of intact, permeabilised or disrupted bacteria in the sludge using a double fluorescent DNA-staining instead of using conventional methods like plate counts and microscope. Physical/mechanical treatments (ultrasonication and high pressure homogenisation) caused moderate effects on cell integrity and caused significant cell disruption only at high specific energy levels. Conversely, thermal treatment caused significant damage of bacterial membranes even at moderate temperatures (45-55 °C). Ozonation significantly affected cell integrity, even at low ozone dosages, below 10 mgO(3)/gTSS, causing an increase of permeabilised and disrupted cells. At higher ozone dosages the compounds solubilised after cell lysis act as scavengers in the competition between soluble compounds and (particulate) bacterial cells. An original aspect of this paper, not yet reported in the literature, is the comparison of the effects of these sludge reduction technologies on bacterial cell integrity and permeabilization by converting pressure, temperature and ozone dosage to an equivalent value of specific energy. Among these technologies, comparison of the applied specific energy demonstrates that achieving the complete disruption of bacterial cells is not always economically advantageous because excessive energy levels may be required. Copyright © 2010 Elsevier Ltd. All rights reserved.

Changes in bacterial diversity associated with bioremediation of used lubricating oil in tropical soils.

PubMed

Meeboon, Naruemon; Leewis, Mary-Cathrine; Kaewsuwan, Sireewan; Maneerat, Suppasil; Leigh, Mary Beth

2017-08-01

Used lubricating oil (ULO) is a widespread contaminant, particularly throughout tropical regions, and may be a candidate for bioremediation. However, little is known about the biodegradation potential or basic microbial ecology of ULO-contaminated soils. This study aims to determine the effects of used ULO on bacterial community structure and diversity. Using a combination of culture-based (agar plate counts) and molecular techniques (16S rRNA gene sequencing and DGGE), we investigated changes in soil bacterial communities from three different ULO-contaminated soils collected from motorcycle mechanical workshops (soil A, B, and C). We further explored the relationship between bacterial community structure, physiochemical soil parameters, and ULO composition in three ULO-contaminated soils. Results indicated that the three investigated soils had different community structures, which may be a result of the different ULO characteristics and physiochemical soil parameters of each site. Soil C had the highest ULO concentration and also the greatest diversity and richness of bacteria, which may be a result of higher nutrient retention, organic matter and cation exchange capacity, as well as freshness of oil compared to the other soils. In soils A and B, Proteobacteria (esp. Gammaproteobacteria) dominated the bacterial community, and in soil C, Actinobacteria and Firmicutes dominated. The genus Enterobacter, a member of the class Gammaproteobacteria, is known to include ULO-degraders, and this genus was the only one found in all three soils, suggesting that it could play a key role in the in situ degradation of ULO-contaminated tropical Thai soils. This study provides insights into our understanding of soil microbial richness, diversity, composition, and structure in tropical ULO-contaminated soils, and may be useful for the development of strategies to improve bioremediation.

Effect of Punica granatum L. Flower Water Extract on Five Common Oral Bacteria and Bacterial Biofilm Formation on Orthodontic Wire

PubMed Central

VAHID DASTJERDI, Elahe; ABDOLAZIMI, Zahra; GHAZANFARIAN, Marzieh; AMDJADI, Parisa; KAMALINEJAD, Mohammad; MAHBOUBI, Arash

2014-01-01

Background: Use of herbal extracts and essences as natural antibacterial compounds has become increasingly popular for the control of oral infectious diseases. Therefore, finding natural antimicrobial products with the lowest side effects seems necessary. The present study sought to assess the effect of Punica granatum L. water extract on five oral bacteria and bacterial biofilm formation on orthodontic wire. Methods: Antibacterial property of P. granatum L. water extract was primarily evaluated in brain heart infusion agar medium using well-plate method. The minimum inhibitory concentration and minimum bactericidal concentration were determined by macro-dilution method. The inhibitory effect on orthodontic wire bacterial biofilm formation was evaluated using viable cell count in biofilm medium. At the final phase, samples were fixed and analyzed by Scanning Electron Microscopy. Results: The growth inhibition zone diameter was proportional to the extract concentration. The water extract demonstrated the maximum antibacterial effect on Streptococcus sanguinis ATCC 10556 with a minimum inhibitory concentration of 6.25 mg/ml and maximum bactericidal effect on S. sanguinis ATCC 10556 and S. sobrinus ATCC 27607 with minimum bactericidal concentration of 25 mg/ml. The water extract decreased bacterial biofilm formation by S. sanguinis, S. sobrinus, S. salivarius, S. mutans ATCC 35608 and E. faecalis CIP 55142 by 93.7–100%, 40.6–99.9%, 85.2–86.5%, 66.4–84.4% and 35.5–56.3% respectively. Conclusion: Punica granatum L. water extract had significant antibacterial properties against 5 oral bacteria and prevented orthodontic wire bacterial biofilm formation. However, further investigations are required to generalize these results to the clinical setting. PMID:26171362

Rapid in situ assessment of physiological activities in bacterial biofilms using fluorescent probes

NASA Technical Reports Server (NTRS)

Yu, F. P.; McFeters, G. A.

1994-01-01

Two rapid in situ enumeration methods using fluorescent probes were used to assess the physiological activities of Klebsiella pneumoniae biofilms on stainless steel. Fluorescent dyes, 5-cyano-2,3-ditolyl tetrazolium chloride (CTC) and rhodamine 123 (Rh 123), were chosen to perform this study. CTC is a soluble redox indicator which can be reduced by respiring bacteria to fluorescent CTC-formazan crystals. Rh 123 is incorporated into bacteria with respect to cellular proton motive force. The intracellular accumulation of these fluorescent dyes can be determined using epifluorescence microscopy. The results obtained with these two fluorescent probes in situ were compared to the plate count (PC) and in situ direct viable count (DVC) methods. Viable cell densities within biofilms determined by the three in situ methods were comparable and always showed approximately 2-fold higher values than those obtained with the PC method. As an additional advantage, the results were observed after 2 h, which was shorter than the 4 h incubation time required for the DVC method and 24 h for colony formation. The results indicate that staining with CTC and Rh 123 provides rapid information regarding cell numbers and physiological activities of bacteria within biofilms.

Bacterial Diversity within the Extreme Arid Atacama Desert Soils of the Yungay Region, Chile

NASA Astrophysics Data System (ADS)

Connon, S. A.; Lester, E. D.; Shafaat, H. S.; Obenhuber, D. C.; Ponce, A.

2006-12-01

Surface and subsurface soil samples analyzed for this study were collected from the hyper-arid Yungay region of the Atacama Desert, Chile. This is the first report of microbial diversity from DNA extracted directly from these extremely desiccated soils. Our data shows that 94% of the 16S rRNA genes cloned from these soils belong to the Actinobacteria phylum. A 24-hour time course series showed a diurnal water activity (aw) cycle that peaked at 0.52 in the early predawn hours, and ranged from 0.08 0.01 during the day. All measured water activity values were below the level required for microbial growth or enzyme activity. Total organic carbon (TOC) levels in this region were just above the limits of detection and ranged from 220 660 μg/g of soil. Phospholipid fatty acid (PLFA) levels indicated cellular biomass ranging from 2 ×105 to 7 ×106 cell equivalents per gram of soil. The culturable counts were low with most samples showing no growth on standard plates of R2A medium; the highest single count was 47 colony forming units (CFU) per gram.

Nonuniform spatial patterns of respiratory activity within biofilms during disinfection.

PubMed Central

Huang, C T; Yu, F P; McFeters, G A; Stewart, P S

1995-01-01

Fluorescent stains in conjunction with cryoembedding and image analysis were applied to demonstrate spatial gradients in respiratory activity within bacterial biofilms during disinfection with monochloramine. Biofilms of Klebsiella pneumoniae and Pseudomonas aeruginosa grown together on stainless steel surfaces in continuous-flow annular reactors were treated with 2 mg of monochloramine per liter (influent concentration) for 2 h. Relatively little biofilm removal occurred as evidenced by total cell direct counts. Plate counts (of both species summed) indicated an average 1.3-log decrease after exposure to 2 mg of monochloramine per liter. The fluorogenic redox indicator 5-cyano-2,3-ditolyl tetrazolium chloride (CTC) and the DNA stain 4',6-diamidino-2-phenylindole (DAPI) were used to differentiate respiring and nonrespiring cells in biofilms. Epifluorescence micrographs of frozen biofilm cross sections clearly revealed gradients of respiratory activity within biofilms in response to monochloramine treatment. These gradients in specific respiratory activity were quantified by calculating the ratio of CTC and DAPI intensities measured by image analysis. Cells near the biofilm-bulk fluid interface lost respiratory activity first. After 2 h of biocide treatment, greater respiratory activity persisted deep in the biofilm than near the biofilm-bulk fluid interface. PMID:7793945

Study of the microbial ecology of wild and aquacultured Tunisian fresh fish.

PubMed

Boulares, Mouna; Mejri, Lobna; Hassouna, Mnasser

2011-10-01

Eighty samples of fresh fish were collected in Tunisia and analyzed for microbial load. Quality and hygienic safety of the meat and intestines of wild and aquacultured fresh fish were determined. The mesophilic aerobic plate count and populations of psychrotrophic lactic acid bacteria (LAB) and other psychrotrophic bacteria ranged from 5.67 to 7.29, 4.51 to 6, and 5.07 to 6.21 log CFU/g, respectively. For all microbiological determinations, bacterial counts were lower in meat than in the intestines of fresh fish. For all samples lower microbial populations were found in most of the wild fish than in the aquacultured fish. No isolates of the pathogenic genera Salmonella and Listeria were detected in any sample. Among the 160 strains of biopreservative psychrotrophic LAB and the 150 strains of spoilage psychrotrophic gram-negative bacteria identified by biochemical and molecular methods, Lactobacillus (six species) and Pseudomonas (six species) predominated. Lactococcus, Leuconostoc, Carnobacterium (C. piscicola and C. divergens), Aeromonas, and Photobacterium were the most common genera, and Lactococcus lactis, Lactobacillus plantarum, Pseudomonas fluorescens, and Aeromonas hydrophila were the most common species. These findings indicate that the microbiological quality of fresh fish in Tunisia can be preserved by controlling pathogenic and psychrotrophic bacteria.

Study on spoilage capability and VBNC state formation and recovery of Lactobacillus plantarum.

PubMed

Liu, Junyan; Li, Lin; Li, Bing; Peters, Brian M; Deng, Yang; Xu, Zhenbo; Shirtliff, Mark E

2017-09-01

The present study aimed at investigating the capability of L. plantarum strain BM-LP14723 to enter into and recover from the viable but nonculturable (VBNC) state and to cause beer spoilage. VBNC state was induced by incubating in beer with subculturing or low temperature treatment. Culturable, total, and viable cells numbers were assessed by MRS agar plate counting, acridine orange direct counting, and Live/Dead BacLight bacterial viability kit, respectively. Organic acids concentrations were measured by reversed-phase high performance liquid chromatography. VBNC L. plantarum cells were detected after 189 ± 1.9 days low temperature treatment or 29 ± 0.7 subcultures in beer. The VBNC L. plantarum retained spoilage capability. Addition of catalase is an effective method for the recovery of the VBNC L. plantarum cells. L. plantarum strain BM-LP14723 is capable of entering into and recovery from the VBNC state and maintained spoilage capability. The current study presented that beer-spoilage L. plantarum can hide both in breweries and during transporting and marketing process and thus lead to beer-spoilage incidents. Copyright © 2017 Elsevier Ltd. All rights reserved.

Rapid Membrane Filtration-Epifluorescent Microscopy Technique for Direct Enumeration of Bacteria in Raw Milk

PubMed Central

Pettipher, Graham L.; Mansell, Roderick; McKinnon, Charles H.; Cousins, Christina M.

1980-01-01

Membrane filtration and epifluorescent microscopy were used for the direct enumeration of bacteria in raw milk. Somatic cells were lysed by treatment with trypsin and Triton X-100 so that 2 ml of milk containing up to 5 × 106 somatic cells/ml could be filtered. The majority of the bacteria (ca. 80%) remained intact and were concentrated on the membrane. After being stained with acridine organe, the bacteria fluoresced under ultraviolet light and could easily be counted. The clump count of orange fluorescing cells on the membrane correlated well (r = 0.91) with the corresponding plate count for farm, tanker, and silo milks. Differences between counts obtained by different operators and between the membrane clump count and plate count were not significant. The technique is rapid, taking less than 25 min, inexpensive, costing less than 50 cents per sample, and is suitable for milks containing 5 × 103 to 5 × 108 bacteria per ml. Images PMID:16345515

Combination of therapeutic ultrasound with antibiotics interfere with the growth of bacterial culture that colonizes skin ulcers: An in-vitro study.

PubMed

Guirro, Elaine Caldeira de Oliveira; Angelis, Dejanira de Franceschi de; Sousa, Natanael Teixeira Alves de; Guirro, Rinaldo Roberto de Jesus

2016-09-01

Staphylococcus aureus and Escherichia coli are among the major bacterial species that colonize skin ulcers. Therapeutic ultrasound (TUS) produces biophysical effects that are relevant to wound healing; however, its application over a contaminated injury is not evidence-based. The objective of this research was to analyze the effect of TUS on in vitro-isolated S. aureus and E. coli, including the combination of ultrasound and antibiotics, in order to assess their antibiotic action on bacterial susceptibility. For the experiments, the bacterial strains were suspended in saline, then diluted (10(4)CFU/mL) for irradiation (at 1 and 3MHz, 0.5 and 0.8W/cm(2) for 0 and 15min) and the combination treatment of ultrasonication and antibiotics was administered by adding nalidixic acid (S. aureus) and tetracycline (E. coli) at concentrations equivalent to 50% of the minimum inhibitory concentration (MIC). The experiments were carried out in duplicate with six repetitions. The suspensions were inoculated on to Petri plates and incubated at 37°C and the colony forming units (CFUs) were counted after 24h. The results were subjected to the Shapiro-Wilk normality test, followed by parametric ANOVA and Tukey's post hoc test at a significance level of 1%. The results demonstrated that the action of TUS at 1MHz inhibited bacterial growth while at 3MHz, bacterial growth was observed in both species. However, the synergistic combination of ultrasound and antibiotics was able to inhibit the growth of both bacteria completely after 15min of ultrasonication. The results suggest that the action of ultrasound on S. aureus and E. coli are dependent on the oscillation frequency as well as the intensity and time of application. The combination of ultrasound with antibiotics was able to inhibit bacterial growth fully at all frequencies and doses in both species. Copyright © 2016 Elsevier B.V. All rights reserved.

Moxifloxacin superior to cefuroxime in reducing bacterial adhesion of Staphylococcus epidermidis on hydrophobic intraocular lenses.

PubMed

Benbouzid, Fathalah; Kodjikian, Laurent; Hartmann, Daniel; Renaud, François; Baillif, Stéphanie

2016-02-01

To compare the anti-adhesive effect of cefuroxime and moxifloxacin on the primary attachment phase of Staphylococcus epidermidis on hydrophobic acrylic intraocular lenses (IOLs). Forty hydrophobic acrylic IOLs were used. Two groups of IOLs were soaked in a moxifloxacin (Mox-T1: 0.5 mg/0.1 ml) or a cefuroxime (Cef-T1: cefuroxime 1 mg/0.1 ml) solution before incubation in a S. epidermidis bacterial suspension. Two other groups were incubated in the bacterial suspension before antibiotics (Cef-T2 and Mox-T2) were added. The control group (Ctrl) consisted of IOLs incubated in the bacterial suspension. After incubation, IOLs were sonicated and vortexed. The resultant suspension was spread over a nutritive agar plate. Bacterial colonies were counted after 24 hr of incubation. Mean number of colony-forming units per IOL was Cef-T1: 184 × 10(3) (SE: 5.24; SD: 28.21), Cef-T2: 117 × 10(3) (SE: 5.74; SD: 30.37), Mox-T1: 1.27 × 10(3) (SE: 0.12; SD: 0.61), Mox-T2: 25 × 10(3) (SE:1.98; SD: 9.72) and Ctrl: 361 × 10(3) (SE: 26.9; SD: 107.6). The number of adhering bacteria did not vary whether cefuroxime was added before or after IOL incubation in the bacterial suspension (p = 0.132). Moxifloxacin was more effective in reducing the number of adhering bacteria when used before IOL incubation (p < 0.001). Overall for T1 and T2, moxifloxacin was more effective than cefuroxime in reducing bacterial adhesion on IOLs (p < 0.001). Moxifloxacin and cefuroxime significantly reduced S. epidermidis adhesion on hydrophobic acrylic IOLs. The anti-adhesive effect was superior with moxifloxacin. © 2015 Acta Ophthalmologica Scandinavica Foundation. Published by John Wiley & Sons Ltd.

The effect of microchannel plate gain depression on PAPA photon counting cameras

NASA Astrophysics Data System (ADS)

Sams, Bruce J., III

1991-03-01

PAPA (precision analog photon address) cameras are photon counting imagers which employ microchannel plates (MCPs) for image intensification. They have been used extensively in astronomical speckle imaging. The PAPA camera can produce artifacts when light incident on its MCP is highly concentrated. The effect is exacerbated by adjusting the strobe detection level too low, so that the camera accepts very small MCP pulses. The artifacts can occur even at low total count rates if the image has highly a concentrated bright spot. This paper describes how to optimize PAPA camera electronics, and describes six techniques which can avoid or minimize addressing errors.

Microbial contamination in intraoral phosphor storage plates: the dilemma.

PubMed

de Souza, Tricia Murielly Pereira Andrade; de Castro, Ricardo Dias; de Vasconcelos, Laís César; Pontual, Andréa Dos Anjos; de Moraes Ramos Perez, Flávia Maria; Pontual, Maria Luiza Dos Anjos

2017-01-01

The aims of this study were to evaluate microbial contamination in phosphor storage plates in dental radiology services and discuss the possible origin of this contamination. The sample comprised 50 phosphor plates: 14 plates from service A, 30 from service B, and 6 in the control group, consisting of plates never used. Damp sterile swabs were rubbed on the phosphor plates, and then transferred to tests tubes containing sterile saline solution. Serial dilutions were made, and then inoculated in triplicate on Mueller Hinton agar plates and incubated at 37 °C/48 h, before counting the colony-forming units (CFU). The samples were also seeded in brain-heart infusion medium to confirm contamination by turbidity of the culture medium. All solutions, turbid and clean, were seeded in selective and non-selective media. At service A and B, 50 and 73.3 % of the phosphor plates were contaminated, respectively. This contamination was mainly due to bacteria of the genus Staphylococcus. CFU counts ranged from 26.4 to 80.0 CFU/plate. Most of the phosphor plates evaluated shown to be contaminated, mainly by Staphylococcus ssp. Quantitatively, this contamination occurred at low levels, possibly arising from handling of the plates. The use of a second plastic barrier may have diminished contamination by microorganisms from the oral cavity. There is a risk of cross-contamination by phosphor storage plates used in dental radiology services.

Evidence of variable bacterial colonization on coloured elastomeric ligatures during orthodontic treatment: An intermodular comparative study.

PubMed

Sharma, Ravish; Sharma, Kavita; Sawhney, Rajesh

2018-03-01

Besides, other factors, the choice of materials used as orthodontic ligatures could be one of the many tools to counter the effects of microbial adhesion, that culminates into dental ailments. Therefore, we assessed bacterial adhesion on elastomeric ligatures with special reference to coloured elastomeric rings during orthodontic treatment. A split mouth study, involving 240 samples of different elastomeric ligatures from forty orthodontic patients possessing good oral hygiene was carried out. The archwire was ligated to the brackets on both arches with elastomeric rings (superslick, clear transparent , blue and pink) at predetermined quadrants. After six weeks, ligatures from second premolars were removed and processed for bacterial enumeration using standard techniques. Bacterial counts were also determined for stimulated saliva samples taken at 0 and 6 weeks. A statistically significant difference in bacterial counts was obtained amongst different elastomeric modules used. Maximum bacterial counts were found on conventional pigmented elastomeric modules, followed by Superslick module and clear module. More number of bacteria associated with the conventional pink as compared to the conventional blue pigmented modules, however it was not statistically significant. The three bacterial genera Streptococcus Staphylococcus and Aerobic Lactobacilli adhered to elastomeric modules in following predominant pattern i.e. Conventional pink>Conventional Blue>Superslick>Clear. The studies evidenced colour and material dependent bacterial colonization on orthodontic modules and could be an indicator of bacterial biofilm forming potential based on surface chemistries and a clinically efficacious tool to redesign conventional and modified elastomeric rings as orthodontic ligation accessories. Key words: Bacterial colonization, biofilm, coloured elastomers, orthodontic ligatures.

[Validation of a clinical prediction rule to distinguish bacterial from aseptic meningitis].

PubMed

Agüero, Gonzalo; Davenport, María C; Del Valle, María de la P; Gallegos, Paulina; Kannemann, Ana L; Bokser, Vivian; Ferrero, Fernando

2010-02-01

Despite most meningitis are not bacterial, antibiotics are usually administered on admission because bacterial meningitis is difficult to be rule-out. Distinguishing bacterial from aseptic meningitis on admission could avoid inappropriate antibiotic use and hospitalization. We aimed to validate a clinical prediction rule to distinguish bacterial from aseptic meningitis in children, on arriving to the emergency room. This prospective study included patients aged < 19 years with meningitis. Cerebrospinal fluid (CSF) and peripheral blood neutrophil count were obtained from all patients. The BMS (Bacterial Meningitis Score) described by Nigrovic (Pediatrics 2002; 110: 712), was calculated: positive CSF Gram stain= 2 points, CSF absolute neutrophil count > or = 1000 cells/mm(3), CSF protein > or = 80 mg/dl, peripheral blood absolute neutrophil count > or = 10.000/mm(3), seizure = 1 point each. Sensitivity (S), specificity (E), positive and negative predictive values (PPV and NPV), positive and negative likelihood ratios (PLR and NLR) of the BMS to predict bacterial meningitis were calculated. Seventy patients with meningitis were included (14 bacterial meningitis). When BMS was calculated, 25 patients showed a BMS= 0 points, 11 BMS= 1 point, and 34 BMS > or = 2 points. A BMS = 0 showed S: 100%, E: 44%, VPP: 31%, VPN: 100%, RVP: 1,81 RVN: 0. A BMS > or = 2 predicted bacterial meningitis with S: 100%, E: 64%, VPP: 41%, VPN: 100%, PLR: 2.8, NLR:0. Using BMS was simple, and allowed identifying children with very low risk of bacterial meningitis. It could be a useful tool to assist clinical decision making.

A New Method for Estimating Bacterial Abundances in Natural Samples using Sublimation

NASA Technical Reports Server (NTRS)

Glavin, Daniel P.; Cleaves, H. James; Schubert, Michael; Aubrey, Andrew; Bada, Jeffrey L.

2004-01-01

We have developed a new method based on the sublimation of adenine from Escherichia coli to estimate bacterial cell counts in natural samples. To demonstrate this technique, several types of natural samples including beach sand, seawater, deep-sea sediment, and two soil samples from the Atacama Desert were heated to a temperature of 500 C for several seconds under reduced pressure. The sublimate was collected on a cold finger and the amount of adenine released from the samples then determined by high performance liquid chromatography (HPLC) with UV absorbance detection. Based on the total amount of adenine recovered from DNA and RNA in these samples, we estimated bacterial cell counts ranging from approx. l0(exp 5) to l0(exp 9) E. coli cell equivalents per gram. For most of these samples, the sublimation based cell counts were in agreement with total bacterial counts obtained by traditional DAPI staining. The simplicity and robustness of the sublimation technique compared to the DAPI staining method makes this approach particularly attractive for use by spacecraft instrumentation. NASA is currently planning to send a lander to Mars in 2009 in order to assess whether or not organic compounds, especially those that might be associated with life, are present in Martian surface samples. Based on our analyses of the Atacama Desert soil samples, several million bacterial cells per gam of Martian soil should be detectable using this sublimation technique.

Introducing a Novel Media to Improve the Recovery of Culturable Bacteria from the Fish Parasite Anisakis spp. larvae (Nematoda: Anisakidae).

PubMed

Svanevik, Cecilie S; Lunestad, Bjørn T

2017-09-01

This paper describes a cultivation method to increase the recovery of bacteria from the marine muscle-invading parasitic nematode larvae of Anisakis spp. These larvae hold a high and complex population of accumulated bacteria, originating from seawater, crustaceans, fish, and marine mammals, all involved in the lifecycle of Anisakis. Two in-house agars based on fish juice prepared by either mechanical or enzymatic degradation of the fish tissue, were made. The Anisakis larvae were homogenised prior to cultivation on the in-house fish juice agars and the bacterial numbers and diversity were compared to those obtained applying the commercially available Marine Agar and Iron Agar Lyngby. Bacterial colonies of unique appearance were subcultured and identified by 16S rRNA gene sequencing. Totally three of twenty identified taxa were found on the in-house fish juice agars only. Fish juice agar prepared enzymatically would be the best supplementary agar, as this agar gave significantly higher heterotrophic plate counts, compared to mechanical preparation. The enzymatically prepared fish juice gave more suitable agar quality, was more resource efficient, and had apparently increased nutrient density and availability.

Salecan Enhances the Activities of β-1,3-Glucanase and Decreases the Biomass of Soil-Borne Fungi

PubMed Central

Chen, Yunmei; Xu, Haiyang; Zhou, Mengyi; Wang, Yang; Wang, Shiming; Zhang, Jianfa

2015-01-01

Salecan, a linear extracellular polysaccharide consisting of β-1,3-D-glucan, has potential applications in the food, pharmaceutical and cosmetic industries. The objective of this study was to evaluate the effects of salecan on soil microbial communities in a vegetable patch. Compositional shifts in the genetic structure of indigenous soil bacterial and fungal communities were monitored using culture-dependent dilution plating, culture-independent PCR-denaturing gradient gel electrophoresis (DGGE) and quantitative PCR. After 60 days, soil microorganism counts showed no significant variation in bacterial density and a marked decrease in the numbers of fungi. The DGGE profiles revealed that salecan changed the composition of the microbial community in soil by increasing the amount of Bacillus strains and decreasing the amount of Fusarium strains. Quantitative PCR confirmed that the populations of the soil-borne fungi Fusarium oxysporum and Trichoderma spp. were decreased approximately 6- and 2-fold, respectively, in soil containing salecan. This decrease in the amount of fungi can be explained by salecan inducing an increase in the activities of β-1,3-glucanase in the soil. These results suggest the promising application of salecan for biological control of pathogens of soil-borne fungi. PMID:26247592

Effects of lead and zinc mining contamination on bacterial community diversity and enzyme activities of vicinal cropland.

PubMed

Qu, Juanjuan; Ren, Guangming; Chen, Bao; Fan, Jinghua; E, Yong

2011-11-01

In the process of mining activity, many kinds of heavy metals enter into soils with dust, causing serious contamination to the environment. In this study, six soils were sampled from cropland at different distances from a lead/zinc mine in Heilongjiang Province, China. The total contents of lead and zinc in the vicinal cropland exceeded the third level of environmental quality standard for soil in China, which indicated that soils in this area were moderately contaminated. Bacterial community diversity and population were greatly decreased when the concentrations of lead and zinc were beyond 1,500 and 995 mg kg(-1), respectively, as analyzed by plate counting and polymerase chain reaction-denaturing gradient gel electrophoresis (PCR-DGGE). The bands of DGGE patterns varied with the degree of contamination. The activities of soil urease, phosphatase, and dehydrogenase were negatively correlated with the concentrations of lead and zinc. The highest inhibitory effect of heavy metals on soil enzyme activities was observed in urease. It was noted that PCR-DGGE patterns combined with soil enzyme activity analysis can be indices for the soil quality assessment by heavy metal contamination.

Response of the Extract-Release Volume and Water-Holding Capacity Phenomena to Microbiologically Spoiled Beef and Aged Beef

PubMed Central

Jay, James M.

1966-01-01

The aging of ground beef was effected by storing in gas-impermeable, sterile plastic bags with incubation at 7 and 15 C. Control meat from the same preparations was wrapped in aluminum foil and stored at the same temperature. In three experiments where control meat was tested, aged meat did not attain a log bacterial number of ca. 8.4 per gram until an average of 6 days after this level was reached in control meats. This degree of difference was shown in values for both extract-release volume (ERV) and water-holding capacity. The previously reported ERV value of around 25, which was found to correspond to an average log bacterial number of ca. 8.5 per gram for ground beef allowed to spoil in aluminum foil and freezer paper, was approximated for aged meats, which required an average of 9.7 days to attain this number compared with 4.1 days for unaged meats. Plate count methods indicated the predominant flora of aged beef to be gram-negative, facultatively psychrophilic rods. Images Fig. 1 Fig. 2 Fig. 4 Fig. 5 PMID:5927019

Comparison of PCR-DGGE and PCR-SSCP analysis for bacterial flora of Japanese traditional fermented fish products, aji-narezushi and iwashi-nukazuke.

PubMed

An, Choa; Takahashi, Hajime; Kimura, Bon; Kuda, Takashi

2010-08-30

The bacterial flora of two Japanese traditional fermented fish products, aji-narezushi (salted and long-fermented horse mackerel (Trachurus japonicas) with rice) and iwashi-nukazuke (salted and long-fermented sardine (Sardinops melanostica) with rice bran), was analysed using non-culture-based polymerase chain reaction (PCR) denaturing gradient gel electrophoresis (DGGE) and culture-based PCR single-strand conformation polymorphism (SSCP) methods. Viable plate counts in aji-narezushi and iwashi-nukazuke were about 6.3-6.6 and 5.7-6.9 log colony-forming units g(-1) respectively. In the PCR-DGGE analysis, Lactobacillus acidipiscis was detected as the predominant bacterium in two of three aji-narezushi samples, while Lactobacillus versmoldensis was predominant in the third sample. By the PCR-SSCP method, Lb. acidipiscis and Lactobacillus plantarum were isolated as the predominant bacteria, while Lb. versmoldensis was not detected. The predominant bacterium in two of three iwashi-nukazuke samples was Tetragenococcus muriaticus, while Tetragenococcus halophilus was predominant in the third sample. The results suggest that the detection of some predominant lactic acid bacteria species in fermented fish by cultivation methods is difficult. Copyright (c) 2010 Society of Chemical Industry.

Microbiology, chemistry and biofilm development in a pilot drinking water distribution system with copper and plastic pipes.

PubMed

Lehtola, Markku J; Miettinen, Ilkka T; Keinänen, Minna M; Kekki, Tomi K; Laine, Olli; Hirvonen, Arja; Vartiainen, Terttu; Martikainen, Pertti J

2004-10-01

We studied the changes in water quality and formation of biofilms occurring in a pilot-scale water distribution system with two generally used pipe materials: copper and plastic (polyethylene, PE). The formation of biofilms with time was analysed as the number of total bacteria, heterotrophic plate counts and the concentration of ATP in biofilms. At the end of the experiment (after 308 days), microbial community structure, viable biomass and gram-negative bacterial biomass were analysed via lipid biomarkers (phospholipid fatty acids and lipopolysaccharide 3-hydroxy fatty acids), and the numbers of virus-like particles and total bacteria were enumerated by SYBR Green I staining. The formation of biofilm was slower in copper pipes than in the PE pipes, but after 200 days there was no difference in microbial numbers between the pipe materials. Copper ion led to lower microbial numbers in water during the first 200 days, but thereafter there were no differences between the two pipe materials. The number of virus-like particles was lower in biofilms and in outlet water from the copper pipes than PE pipes. Pipe material influenced also the microbial and gram-negative bacterial community structure in biofilms and water.

[USE OF LASER RAMAN-LUMINESCENT TECHNOLOGIES FOR EVALUATION OF QUALITY OF MEAT PRODUCTS AND DETERMINATION OF THE DEGREE OF THEIR BACTERIAL CONTAMINATION].

PubMed

Kukushkin, V I; Satusheva, E V; Aleksandrov, M T; Morozova, O A; Pashkov, E P; Ambartsumyan, O A; Amosova, V A

2015-01-01

Determination of the effect of microorganisms on spoilage of meat products during various temperature regimes of storage by integral indexes of luminescent lines in their spectra and development of an algorithm of microorganism indication by an express method using laser Raman-luminescent spectroscopy. Minced meat from beef and pork was used. Determination of quantity of mesophilic aerobic and opportunistic-anaerobic microorganisms was carried out by serial 10-fold dilutions with subsequent parallel seeding into Rida count total 24 plates and Petri dishes with 5% blood agar. Sample study was carried out in luminescent software-hardware complex Enspectr L405 (a variant of Enspectr M software-hardware complexes). Meat spoilage was established to be caused to a large degree by Pseudomonas genus (P. fluorescens, P. putida, P. fragi et al.) bacterial growth. Raman-luminescent spectra of bacteria that compose microflora, characterizing and accompanying beef and pork spoilage, were measured and recorded into a database. The results obtained will allow to use this technique in the future for both express-indication and differentiation of microorganisms and express-evaluation of quality of meat products at all stages of their manufacturing, storage, transport and realization.

Portable bacterial identification system based on elastic light scatter patterns.

PubMed

Bae, Euiwon; Ying, Dawei; Kramer, Donald; Patsekin, Valery; Rajwa, Bartek; Holdman, Cheryl; Sturgis, Jennifer; Davisson, V Jo; Robinson, J Paul

2012-08-28

Conventional diagnosis and identification of bacteria requires shipment of samples to a laboratory for genetic and biochemical analysis. This process can take days and imposes significant delay to action in situations where timely intervention can save lives and reduce associated costs. To enable faster response to an outbreak, a low-cost, small-footprint, portable microbial-identification instrument using forward scatterometry has been developed. This device, weighing 9 lb and measuring 12 × 6 × 10.5 in., utilizes elastic light scatter (ELS) patterns to accurately capture bacterial colony characteristics and delivers the classification results via wireless access. The overall system consists of two CCD cameras, one rotational and one translational stage, and a 635-nm laser diode. Various software algorithms such as Hough transform, 2-D geometric moments, and the traveling salesman problem (TSP) have been implemented to provide colony count and circularity, centering process, and minimized travel time among colonies. Experiments were conducted with four bacteria genera using pure and mixed plate and as proof of principle a field test was conducted in four different locations where the average classification rate ranged between 95 and 100%.

Phylogenetic changes in soil microbial and diazotrophic diversity with application of butachlor.

PubMed

Yen, Jui-Hung; Wang, Yei-Shung; Hsu, Wey-Shin; Chen, Wen-Ching

2013-01-01

We investigated changes in population and taxonomic distribution of cultivable bacteria and diazotrophs with butachlor application in rice paddy soils. Population changes were measured by the traditional plate-count method, and taxonomic distribution was studied by 16S rDNA sequencing, then maximum parsimony phylogenic analysis with bootstrapping (1,000 replications). The bacterial population was higher after 39 than 7 days of rice cultivation, which indicated the augmentation of soil microbes by rice root exudates. The application of butachlor increased the diazotrophic population in both upper (0-3 cm) and lower (3-15 cm) layers of soils. Especially at day 39, the population of diazotrophs was 1.8 and 1.6 times that of the control in upper and lower layer soils, respectively. We found several bacterial strains only with butachlor application; examples are strains closest to Bacillus arsenicus, B. marisflavi, B. luciferensis, B. pumilus, and Pseudomonas alvei. Among diazotrophs, three strains closely related to Streptomyces sp. or Rhrizobium sp. were found only with butachlor application. The population of cultivable bacteria and the species composition were both changed with butachlor application, which explains in part the contribution of butachlor to augmenting soil nitrogen-fixing ability.

Evolution of bacterial communities in the Gironde Estuary (France) according to a salinity gradient

NASA Astrophysics Data System (ADS)

Prieur, D.; Troussellier, M.; Romana, A.; Chamroux, S.; Mevel, G.; Baleux, B.

1987-01-01

Three surveys were performed in the Gironde Estuary (France) in August 1981, March 1982 and July 1982. For each campaign, seventy samples were taken by helicopter, in order to follow the tide along the estuary. Of the parameters that were studied, salinity appeared to be the most important and which controls the bacterial communities along the estuary. This paper deals with the evolution of bacterial communities along a salinity gradient. The information obtained from various bacteriological parameters (total bacterial counts, viable counts on salted and unsalted media, functional evenness) were convergent. The bacterial community is dominated by an halotolerant microflora. In the estuary, a continental microflora is followed by a marine microflora. The succession zone between these two microflora is located between 5 and 10‰ areas of salinity.

Intermittent fasting promotes bacterial clearance and intestinal IgA production in Salmonella typhimurium-infected mice.

PubMed

Godínez-Victoria, M; Campos-Rodriguez, R; Rivera-Aguilar, V; Lara-Padilla, E; Pacheco-Yepez, J; Jarillo-Luna, R A; Drago-Serrano, M E

2014-05-01

The impact of intermittent fasting versus ad libitum feeding during Salmonella typhimurium infection was evaluated in terms of duodenum IgA levels, bacterial clearance and intestinal and extra-intestinal infection susceptibility. Mice that were intermittently fasted for 12 weeks or fed ad libitum were infected with S. typhimurium and assessed at 7 and 14 days post-infection. Next, we evaluated bacterial load in the faeces, Peyer's patches, spleen and liver by plate counting, as well as total and specific intestinal IgA and plasmatic corticosterone levels (by immunoenzymatic assay) and lamina propria IgA levels in plasma cells (by cytofluorometry). Polymeric immunoglobulin receptor, α- and J-chains, Pax-5 factor, pro-inflammatory cytokine (tumour necrosis factor-α and interferon-γ) and anti-inflammatory cytokine (transforming growth factor-β) mRNA levels were assessed in mucosal and liver samples (by real-time PCR). Compared with the infected ad libitum mice, the intermittently fasted infected animals had (1) lower intestinal and systemic bacterial loads; (2) higher SIgA and IgA plasma cell levels; (3) higher mRNA expression of most intestinal parameters; and (4) increased or decreased corticosterone levels on day 7 and 14 post-infection, respectively. No contribution of liver IgA was observed at the intestinal level. Apparently, the changes following metabolic stress induced by intermittent fasting during food deprivation days increased the resistance to S. typhimurium infection by triggering intestinal IgA production and presumably, pathogen elimination by phagocytic inflammatory cells. © 2014 John Wiley & Sons Ltd.

Covalent Immobilization of Enoxacin onto Titanium Implant Surfaces for Inhibiting Multiple Bacterial Species Infection and In Vivo Methicillin-Resistant Staphylococcus aureus Infection Prophylaxis

PubMed Central

Nie, Bin'en; Long, Teng; Ao, Haiyong; Zhou, Jianliang; Tang, Tingting

2016-01-01

ABSTRACT Infection is one of the most important causes of titanium implant failure in vivo. A developing prophylactic method involves the immobilization of antibiotics, especially vancomycin, onto the surface of the titanium implant. However, these methods have a limited effect in curbing multiple bacterial infections due to antibiotic specificity. In the current study, enoxacin was covalently bound to an amine-functionalized Ti surface by use of a polyethylene glycol (PEG) spacer, and the bactericidal effectiveness was investigated in vitro and in vivo. The titanium surface was amine functionalized with 3-aminopropyltriethoxysilane (APTES), through which PEG spacer molecules were covalently immobilized onto the titanium, and then the enoxacin was covalently bound to the PEG, which was confirmed by X-ray photoelectron spectrometry (XPS). A spread plate assay, confocal laser scanning microscopy (CLSM), and scanning electron microscopy (SEM) were used to characterize the antimicrobial activity. For the in vivo study, Ti implants were inoculated with methicillin-resistant Staphylococcus aureus (MRSA) and implanted into the femoral medullary cavity of rats. The degree of infection was assessed by radiography, micro-computed tomography, and determination of the counts of adherent bacteria 3 weeks after surgery. Our data demonstrate that the enoxacin-modified PEGylated Ti surface effectively prevented bacterial colonization without compromising cell viability, adhesion, or proliferation in vitro. Furthermore, it prevented MRSA infection of the Ti implants in vivo. Taken together, our results demonstrate that the use of enoxacin-modified Ti is a potential approach to the alleviation of infections of Ti implants by multiple bacterial species. PMID:27799220

Surface charge-conversion polymeric nanoparticles for photodynamic treatment of urinary tract bacterial infections

NASA Astrophysics Data System (ADS)

Liu, Shijie; Qiao, Shenglin; Li, Lili; Qi, Guobin; Lin, Yaoxin; Qiao, Zengying; Wang, Hao; Shao, Chen

2015-12-01

Urinary tract infections are typical bacterial infections which result in a number of economic burdens. With increasing antibiotic resistance, it is urgent that new approaches are explored that can eliminate pathogenic bacteria without inducing drug resistance. Antimicrobial photodynamic therapy (PDT) is a new promising tactic. It is a gentle in situ photochemical reaction in which a photosensitizer (PS) generates reactive oxygen species (ROS) under laser irradiation. In this work, we have demonstrated Chlorin e6 (Ce6) encapsulated charge-conversion polymeric nanoparticles (NPs) for efficiently targeting and killing pathogenic bacteria in a weakly acidic urinary tract infection environment. Owing to the surface charge conversion of NPs in an acidic environment, the NPs exhibited enhanced recognition for Gram-positive (ex. S. aureus) and Gram-negative (ex. E. coli) bacteria due to the charge interaction. Also, those NPs showed significant antibacterial efficacy in vitro with low cytotoxicity. The MIC value of NPs to E. coli is 17.91 μg ml-1, compared with the free Ce6 value of 29.85 μg ml-1. Finally, a mouse acute cystitis model was used to assess the photodynamic therapy effects in urinary tract infections. A significant decline (P < 0.05) in bacterial cells between NPs and free Ce6 occurred in urine after photodynamic therapy treatment. And the plated counting results revealed a remarkable bacterial cells drop (P < 0.05) in the sacrificed bladder tissue. Above all, this nanotechnology strategy opens a new door for the treatment of urinary tract infections with minimal side effects.

Dynamics of bacterial communities before and after distribution in a full-scale drinking water network.

PubMed

El-Chakhtoura, Joline; Prest, Emmanuelle; Saikaly, Pascal; van Loosdrecht, Mark; Hammes, Frederik; Vrouwenvelder, Hans

2015-05-01

Understanding the biological stability of drinking water distribution systems is imperative in the framework of process control and risk management. The objective of this research was to examine the dynamics of the bacterial community during drinking water distribution at high temporal resolution. Water samples (156 in total) were collected over short time-scales (minutes/hours/days) from the outlet of a treatment plant and a location in its corresponding distribution network. The drinking water is treated by biofiltration and disinfectant residuals are absent during distribution. The community was analyzed by 16S rRNA gene pyrosequencing and flow cytometry as well as conventional, culture-based methods. Despite a random dramatic event (detected with pyrosequencing and flow cytometry but not with plate counts), the bacterial community profile at the two locations did not vary significantly over time. A diverse core microbiome was shared between the two locations (58-65% of the taxa and 86-91% of the sequences) and found to be dependent on the treatment strategy. The bacterial community structure changed during distribution, with greater richness detected in the network and phyla such as Acidobacteria and Gemmatimonadetes becoming abundant. The rare taxa displayed the highest dynamicity, causing the major change during water distribution. This change did not have hygienic implications and is contingent on the sensitivity of the applied methods. The concept of biological stability therefore needs to be revised. Biostability is generally desired in drinking water guidelines but may be difficult to achieve in large-scale complex distribution systems that are inherently dynamic. Copyright © 2015 Elsevier Ltd. All rights reserved.

Surface charge-conversion polymeric nanoparticles for photodynamic treatment of urinary tract bacterial infections.

PubMed

Liu, Shijie; Qiao, Shenglin; Li, Lili; Qi, Guobin; Lin, Yaoxin; Qiao, Zengying; Wang, Hao; Shao, Chen

2015-12-11

Urinary tract infections are typical bacterial infections which result in a number of economic burdens. With increasing antibiotic resistance, it is urgent that new approaches are explored that can eliminate pathogenic bacteria without inducing drug resistance. Antimicrobial photodynamic therapy (PDT) is a new promising tactic. It is a gentle in situ photochemical reaction in which a photosensitizer (PS) generates reactive oxygen species (ROS) under laser irradiation. In this work, we have demonstrated Chlorin e6 (Ce6) encapsulated charge-conversion polymeric nanoparticles (NPs) for efficiently targeting and killing pathogenic bacteria in a weakly acidic urinary tract infection environment. Owing to the surface charge conversion of NPs in an acidic environment, the NPs exhibited enhanced recognition for Gram-positive (ex. S. aureus) and Gram-negative (ex. E. coli) bacteria due to the charge interaction. Also, those NPs showed significant antibacterial efficacy in vitro with low cytotoxicity. The MIC value of NPs to E. coli is 17.91 μg ml(-1), compared with the free Ce6 value of 29.85 μg ml(-1). Finally, a mouse acute cystitis model was used to assess the photodynamic therapy effects in urinary tract infections. A significant decline (P < 0.05) in bacterial cells between NPs and free Ce6 occurred in urine after photodynamic therapy treatment. And the plated counting results revealed a remarkable bacterial cells drop (P < 0.05) in the sacrificed bladder tissue. Above all, this nanotechnology strategy opens a new door for the treatment of urinary tract infections with minimal side effects.

Comparison of wiping and rinsing techniques after oral care procedures in critically ill patients during endotracheal intubation and after extubation: A prospective cross-over trial.

PubMed

Muramatsu, Keita; Matsuo, Koichiro; Kawai, Yusuke; Yamamoto, Tsukasa; Hara, Yoshitaka; Shimomura, Yasuyo; Yamashita, Chizuru; Nishida, Osamu

2018-06-26

Endotracheal intubation of critically ill patients increases the risk of aspiration pneumonia, which can be reduced by regular oral care. However, the rinsing of the residual oral contaminants after mechanical cleaning carries the risk of aspirating the residue during the intubation period. Removing the contaminants by wiping with mouth wipes could be an alternative to rinsing with water because of no additional fluid. This study tested: (i) the amount of oral bacteria during endotracheal intubation and after extubation; and (ii) the changes in the bacterial count during oral care procedures. Thirty-five mechanically ventilated patients in the intensive care unit were enrolled. The amount of bacteria on the dorsal tongue surface was counted before and following oral care and then after the elimination of contaminants either by rinsing with water and suctioning or by wiping with mouth wipes. The oral bacterial amount was compared statistically between the intubation and extubation status and among set time points during the oral care procedure. The oral bacterial count was significantly decreased after extubation. During the oral care procedure, the oral bacterial amount was significantly lower after eliminating the contaminants either by rinsing or wiping, with no remarkable difference between the elimination techniques. The findings suggest that the oral bacterial amount is elevated during endotracheal intubation, which could increase the risk of aspiration pneumonia. The significant reduction in the bacterial count by wiping indicates that it might be a suitable alternative to rinsing for mechanically ventilated patients. © 2018 Japan Academy of Nursing Science.

Flow cytometry analysis using sysmex UF-1000i classifies uropathogens based on bacterial, leukocyte, and erythrocyte counts in urine specimens among patients with urinary tract infections.

PubMed

Monsen, Tor; Rydén, Patrik

2015-02-01

Urinary tract infections (UTIs) are the second most common bacterial infection. Urine culture is the gold standard for diagnosis, but new techniques, such as flow cytometry analysis (FCA), have been introduced. The aim of the present study was to evaluate FCA characteristics regarding bacteriuria, leukocyturia, and erythrocyturia in relation to cultured uropathogens in specimens from patients with a suspected UTI. We also wanted to evaluate whether the FCA characteristics can identify uropathogens prior to culture. From a prospective study, 1,587 consecutive urine specimens underwent FCA prior to culture during January and February 2012. Outpatients and inpatients (79.6% and 19.4%, respectively) were included, of whom women represented 67.5%. In total, 620 specimens yielded growth, of which Escherichia coli represented 65%, Enterococcus spp. 8%, Klebsiella spp. 7%, and Staphylococcus spp. 5%. For the uropathogens, the outcome of FCA was compared against the results for specimens with E. coli and those with a negative culture. E. coli had high bacterial (median, 17,914/μl), leukocyte (median, 348/μl), and erythrocyte (median, 23/μl) counts. With the exception of Klebsiella spp., the majority of the uropathogens had considerable or significantly lower bacterial counts than that of E. coli. High leukocyte counts were found in specimens with Staphylococcus aureus, Proteus mirabilis, Pseudomonas aeruginosa, and group C streptococci. Elevated erythrocyte counts were found for P. vulgaris, P. aeruginosa, and group C streptococci, as well as for Staphylococcus saprophyticus. In essence, FCA adds new information about the bacterial, leukocyte, and erythrocyte counts in urine specimens for different uropathogens. Based on FCA characteristics, uropathogens can be classified and identified prior to culture. E. coli and Klebsiella spp. have similar FCA characteristics. Copyright © 2015, American Society for Microbiology. All Rights Reserved.

Gingival fluid cytokine expression and subgingival bacterial counts during pregnancy and postpartum: a case series.

PubMed

Bieri, Regina Alessandri; Adriaens, Laurence; Spörri, Stefan; Lang, Niklaus P; Persson, G Rutger

2013-01-01

The aim of this study was to assess gingival fluid (GCF) cytokine messenger RNA (mRNA) levels, subgingival bacteria, and clinical periodontal conditions during a normal pregnancy to postpartum. Subgingival bacterial samples were analyzed with the checkerboard DNA-DNA hybridization method. GCF samples were assessed with real-time PCR including five proinflammatory cytokines and secretory leukocyte protease inhibitor. Nineteen pregnant women with a mean age of 32 years (S.D. ± 4 years, range 26-42) participated in the study. Full-mouth bleeding scores (BOP) decreased from an average of 41.2% (S.D. ± 18.6%) at the 12th week of pregnancy to 26.6% (S.D. ± 14.4%) at the 4-6 weeks postpartum (p < 0.001). Between week 12 and 4-6 weeks postpartum, the mean probing pocket depth changed from 2.4 mm (S.D. ± 0.4) to 2.3 mm (S.D. ± 0.3) (p = 0.34). Higher counts of Eubacterium saburreum, Parvimonas micra, Selenomonas noxia, and Staphylococcus aureus were found at week 12 of pregnancy than at the 4-6 weeks postpartum examinations (p < 0.001). During and after pregnancy, statistically significant correlations between BOP scores and bacterial counts were observed. BOP scores and GCF levels of selected cytokines were not related to each other and no differences in GCF levels of the cytokines were observed between samples from the 12th week of pregnancy to 4-6 weeks postpartum. Decreasing postpartum counts of Porphyromonas endodontalis and Pseudomonas aeruginosa were associated with decreasing levels of Il-8 and Il-1β. BOP decreased after pregnancy without any active periodontal therapy. Associations between bacterial counts and cytokine levels varied greatly in pregnant women with gingivitis and a normal pregnancy outcome. Postpartum associations between GCF cytokines and bacterial counts were more consistent. Combined assessments of gingival fluid cytokines and subgingival bacteria may provide important information on host response.

Microbial colonization at the implant-abutment interface and its possible influence on periimplantitis: A systematic review and meta-analysis.

PubMed

Tallarico, Marco; Canullo, Luigi; Caneva, Martina; Özcan, Mutlu

2017-07-01

The aim of this systematic review and meta-analysis was to evaluate the microbial colonization at the implant-abutment interfaces (IAI) on bone-level implants and to identify possible association with peri-implant conditions. The focus question aimed to answer whether two-piece osseointegrated implants, in function for at least 1 year, in human, relate to higher bacterial count and the onset of periimplantitis, compared to healthy peri-implant conditions. Search strategy encompassed the on-line (MedLine, Google scholar, Cochrane library) literature from 1990 up to March 2015 published in English using combinations of MeSH (Medical Subject Headings) and search terms. Quality assessment of selected full-text articles was performed according to the ARRIVE and CONSORT statement guidelines. For data analysis, the total bacterial count of Porphyromonas gingivalis, Tannerella forsythia, Treponema denticola, Prevotella intermedia, and Fusobacterium nucleatum was calculated and compared to IAI with or without peri-implant pathology. A total of 14 articles, reporting data from 1126 implants, fulfilled the inclusion criteria and subjected to quality assessment. The selected studies revealed contamination of the IAI, in patients who received two-piece implant systems. Meta-analysis indicated significant difference in total bacterial count between implants affected by periimplantitis versus healthy peri-implant tissues (0.387±0.055; 95% CI 0.279-0.496). Less bacterial counts were identified in the healthy IAI for all the investigated gram-negative bacteria except for T. forsythia. Significantly higher bacterial counts were found for periodontal pathogenic bacteria within the IAI of implants in patients with periimplantitis compared to those implants surrounded by healthy peri-implant tissues. Copyright © 2017 Japan Prosthodontic Society. Published by Elsevier Ltd. All rights reserved.

In situ evaluation of a new silorane-based composite resin's bioadhesion properties.

PubMed

Claro-Pereira, Diogo; Sampaio-Maia, Benedita; Ferreira, Carla; Rodrigues, Andreia; Melo, Luís F; Vasconcelos, Mário R

2011-12-01

The aim of the present study was to compare, in situ, the initial dental plaque formation on a recently developed silorane-based composite resin, Filtek Silorane, and on a widely used methacrylate-based composite resin, Synergy D6, and to relate possible differences to surface free energy, hydrophobicity and type of organic matrix. Discs of Filtek Silorane and Synergy D6 were prepared and polished equally in order to attain the same surface roughness. Water, formamide and 1-bromonaphthalene contact angles were determined and the surface free energy and the hydrophobicity of the materials calculated. Two discs of each material were mounted in individual oral splints and exposed to the oral cavity of 20 participants for 4h. After this period the microbial adhesion to both materials' surface was measured by two different approaches, the DAPI staining and the plate count. Statistical analysis was performed using non-parametric tests. The surface roughness (R(a) parameter) was similar between the two materials and lower than 0.2μm. Mean water and formamide contact angles were significantly higher for Filtek Silorane, which presented significantly lower surface free energy and greater degree of hydrophobicity in comparison to Synergy D6. The bioadhesion potential evaluated by either DAPI staining or plate count did not differ between the two materials. In contrast to previous in vitro studies, the present in situ study found no statistically significant differences with respect to bacterial adhesion between Filtek Silorane and Synergy D6, despite the differences found for surface free energy and hydrophobicity. Copyright © 2011 Academy of Dental Materials. Published by Elsevier Ltd. All rights reserved.

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.

Assessment of pathogenic bacteria in water and sediment from a water reservoir under tropical conditions (Lake Ma Vallée), Kinshasa Democratic Republic of Congo.

PubMed

Mwanamoki, Paola M; Devarajan, Naresh; Thevenon, Florian; Atibu, Emmanuel K; Tshibanda, Joseph B; Ngelinkoto, Patience; Mpiana, Pius T; Prabakar, Kandasamy; Mubedi, Josué I; Kabele, Christophe G; Wildi, Walter; Poté, John

2014-10-01

This study was conducted to assess potential human health risks presented by pathogenic bacteria in a protected multi-use lake-reservoir (Lake Ma Vallée) located in west of Kinshasa, Democratic Republic of Congo (DRC). Water and surface sediments from several points of the Lake were collected during summer. Microbial analysis was performed for Escherichia coli, Enterococcus (ENT), Pseudomonas species and heterotrophic plate counts. PCR amplification was performed for the confirmation of E. coli, ENT, Pseudomonas spp. and Pseudomonas aeruginosa isolated from samples. The results reveal low concentration of bacteria in water column of the lake, the bacterial quantification results observed in this study for the water column were below the recommended limits, according to WHO and the European Directive 2006/7/CE, for bathing water. However, high concentration of bacteria was observed in the sediment samples; the values of 2.65 × 10(3), 6.35 × 10(3), 3.27 × 10(3) and 3.60 × 10(8) CFU g(-1) of dry sediment for E. coli, ENT, Pseudomonas spp. and heterotrophic plate counts, respectively. The results of this study indicate that sediments of the Lake Ma Vallée can constitute a reservoir of pathogenic microorganisms which can persist in the lake. Possible resuspension of faecal indicator bacteria and pathogens would affect water quality and may increase health risks to the population during recreational activities. Our results indicate that the microbial sediment analysis provides complementary and important information for assessing sanitary quality of surface water under tropical conditions.

Evaluation of the limulus amoebocyte lysate test in conjunction with a gram negative bacterial plate count for detecting irradiation of chicken

NASA Astrophysics Data System (ADS)

Scotter, Susan L.; Wood, Roger; McWeeny, David J.

A study to evaluate the potential of the Limulus amoebocyte lysate (LAL) test in conjuction with a Gram negative bacteria (GNB) plate count for detecting the irradiation of chicken is described. Preliminary studies demonstrated that chickens irradiated at an absorbed dose of 2.5 kGy could be differentiated from unirradiated birds by measuring levels of endotoxin and of numbers of GNB on chicken skin. Irradiated birds were found to have endotoxin levels similar to those found in unirradiated birds but significantly lower numbers of GNB. In a limited study the test was found to be applicable to birds from different processors. The effect of temperature abuse on the microbiological profile, and thus the efficacy of the test, was also investigated. After temperature abuse, the irradiated birds were identifiable at worst up to 3 days after irradiation treatment at the 2.5 kGy level and at best some 13 days after irradiation. Temperature abuse at 15°C resulted in rapid recovery of surviving micro-organisms which made differentiation of irradiated and unirradiated birds using this test unreliable. The microbiological quality of the bird prior to irradiation treatment also affected the test as large numbers of GNB present on the bird prior to irradiation treatment resulted in larger numbers of survivors. In addition, monitoring the developing flora after irradiation treatment and during subsequent chilled storage also aided differentiation of irradiated and unirradiated birds. Large numbers of yeasts and Gram positive cocci were isolated from irradiated carcasses whereas Gram negative oxidative rods were the predominant spoilage flora on unirradiated birds.

Serum bactericidal assay for the evaluation of typhoid vaccine using a semi-automated colony-counting method.

PubMed

Jang, Mi Seon; Sahastrabuddhe, Sushant; Yun, Cheol-Heui; Han, Seung Hyun; Yang, Jae Seung

2016-08-01

Typhoid fever, mainly caused by Salmonella enterica serovar Typhi (S. Typhi), is a life-threatening disease, mostly in developing countries. Enzyme-linked immunosorbent assay (ELISA) is widely used to quantify antibodies against S. Typhi in serum but does not provide information about functional antibody titers. Although the serum bactericidal assay (SBA) using an agar plate is often used to measure functional antibody titers against various bacterial pathogens in clinical specimens, it has rarely been used for typhoid vaccines because it is time-consuming and labor-intensive. In the present study, we established an improved SBA against S. Typhi using a semi-automated colony-counting system with a square agar plate harboring 24 samples. The semi-automated SBA efficiently measured bactericidal titers of sera from individuals immunized with S. Typhi Vi polysaccharide vaccines. The assay specifically responded to S. Typhi Ty2 but not to other irrelevant enteric bacteria including Vibrio cholerae and Shigella flexneri. Baby rabbit complement was more appropriate source for the SBA against S. Typhi than complements from adult rabbit, guinea pig, and human. We also examined the correlation between SBA and ELISA for measuring antibody responses against S. Typhi using pre- and post-vaccination sera from 18 human volunteers. The SBA titer showed a good correlation with anti-Vi IgG quantity in the serum as determined by Spearman correlation coefficient of 0.737 (P < 0.001). Taken together, the semi-automated SBA might be efficient, accurate, sensitive, and specific enough to measure functional antibody titers against S. Typhi in sera from human subjects immunized with typhoid vaccines. Copyright © 2016 The Authors. Published by Elsevier Ltd.. All rights reserved.

Antibacterial Activities of Hibiscus sabdariffa Extracts and Chemical Sanitizers Directly on Green Leaves Contaminated with Foodborne Pathogens.

PubMed

Gómez-Aldapa, Carlos A; Rangel-Vargas, Esmeralda; Torres-Vitela, Ma Refugio; Villarruel-López, Angélica; Acevedo-Sandoval, Otilio A; Gordillo-Martínez, Alberto J; Godínez-Oviedo, Angélica; Castro-Rosas, Javier

2018-02-01

Leafy greens have been associated with foodborne disease outbreaks in different countries. To decrease microbial contamination of leafy greens, chemical agents are commonly used; however, a number of studies have shown these agents to have limited antimicrobial effect against pathogenic bacteria on vegetables. The objective of this study was to compare the antibacterial effect of Hibiscus sabdariffa calyx extracts (water, methanol, acetone, and ethyl acetate), sodium hypochlorite, acetic acid, and colloidal silver against foodborne bacteria on leafy greens. Thirteen foodborne bacteria were used in the study: Listeria monocytogenes, Shigella flexneri, Salmonella serotypes Typhimurium Typhi, and Montevideo, Staphylococcus aureus, Escherichia coli O157:H7, five E. coli pathotypes (Shiga toxin-producing, enteropathogenic, enterotoxigenic, enteroinvasive, and enteroaggregative), and Vibrio cholerae O1. Each foodborne bacterium was separately inoculated on romaine lettuce, spinach, and coriander leaves. Separately, contaminated leafy greens were immersed in four hibiscus extracts and in sanitizers for 5 min. Next, green leaves were washed with sterile tap water. Separately, each green leaf was placed in a bag that contained 0.1% sterile peptone water and was rubbed for 2 min. Counts were done by plate count using appropriate dilutions (in sterile peptone water) of the bacterial suspensions spread on Trypticase soy agar plates and incubated at 35 ± 2°C for 48 h. Statistically significant differences ( P < 0.05) were calculated with an analysis of variance and Duncan's test. All 13 foodborne bacteria attached to leafy greens. Roselle calyx extracts caused a significantly greater reduction ( P < 0.05) in concentration of all foodborne bacteria on contaminated romaine lettuce, spinach, and coriander than did the sodium hypochlorite, colloidal silver, and acetic acid. Dry roselle calyx extracts may potentially be a useful addition to disinfection procedures for romaine lettuce, spinach, and coriander.

Development of multiplex PCR for the detection of total coliform bacteria for Escherichia coli and Clostridium perfringens in drinking water.

PubMed

Tantawiwat, Suwalee; Tansuphasiri, Unchalee; Wongwit, Waranya; Wongchotigul, Varee; Kitayaporn, Dwip

2005-01-01

Multiplex PCR amplification of lacZ, uidA and plc genes was developed for the simultaneous detection of total coliform bacteria for Escherichia coli and Clostridium perfringens, in drinking water. Detection by agarose gel electrophoresis yielded a band of 876 bp for the lacZ gene of all coliform bacteria; a band of 147 bp for the uidA gene and a band of 876 bp for the lacZ gene of all strains of E. coli; a band of 280 bp for the p/c gene for all strains of C. perfringens; and a negative result for all three genes when tested with other bacteria. The detection limit was 100 pg for E. coli and C. perfringens, and 1 ng for coliform bacteria when measured with purified DNA. This assay was applied to the detection of these bacteria in spiked water samples. Spiked water samples with 0-1,000 CFU/ml of coliform bacteria and/or E. coli and/or C. perfringens were detected by this multiplex PCR after a pre-enrichment step to increase the sensitivity and to ensure that the detection was based on the presence of cultivable bacteria. The result of bacterial detection from the multiplex PCR was comparable with that of a standard plate count on selective medium (p=0.62). When using standard plate counts as a gold standard, the sensitivity for this test was 99.1% (95% CI 95.33, 99.98) and the specificity was 90.9 % (95% CI 75.67, 98.08). Multiplex PCR amplification with a pre-enrichment step was shown to be an effective, sensitive and rapid method for the simultaneous detection of these three microbiological parameters in drinking water.

Heterotrophic plate count and consumer's health under special consideration of water softeners.

PubMed

Hambsch, Beate; Sacré, Clara; Wagner, Ivo

2004-05-01

The phenomenon of bacterial growth in water softeners is well known since years. To upgrade the hygienic safety of water softeners, the German DIN Standard 19636 was developed, to assure that the distribution system could not be contaminated by these devices and that the drinking water to be used in the household still meets the microbiological standards according to the German drinking water guidelines, i.e. among others heterotrophic plate count (HPC) below 100 CFU/ml. Moreover, the standard for the water softeners includes a test for contamination with Pseudomonas aeruginosa which has to be disinfected during the regeneration phase. This is possible by sanitizing the resin bed during regeneration by producing chlorine. The results of the last 10 years of tests of water softeners according to DIN 19636 showed that it is possible to produce water softeners that comply with that standard. Approximately 60% of the tested models were accepted. P. aeruginosa is used as an indicator for potentially pathogenic bacteria being able to grow also in low nutrient conditions which normally prevail in drinking water. Like other heterotrophs, the numbers of P. aeruginosa increase rapidly as stagnation occurs. Normally P. aeruginosa is not present in the distributed drinking water. However, under certain conditions, P. aeruginosa can be introduced into the drinking water distribution system, for instance, during construction work. The occurrence of P. aeruginosa is shown in different cases in treatment plants, public drinking water systems and in-house installations. The compliance with DIN 19636 provides assurance that a water softener will not be a constant source of contamination, even if it is once inoculated with a potentially pathogenic bacterium like P. aeruginosa. Copyright 2003 Elsevier B.V.

Simplifying Collection of Corneal Specimens in Cases of Suspected Bacterial Keratitis

PubMed Central

Kaye, Stephen B.; Rao, Prasad G.; Smith, Godfrey; Scott, John A.; Hoyles, Sharon; Morton, Clare E.; Willoughby, Colin; Batterbury, Mark; Harvey, Graham

2003-01-01

Identification of the causative organisms in suspected bacterial keratitis traditionally involves collecting multiple corneal scrapes, which are plated directly onto different solid agar culture media. Difficulties have been reported with this practice, so the development of a simpler diagnostic method in suspected bacterial keratitis would be useful. It is unclear whether a single corneal scrape sent to the microbiology laboratory in a liquid transport culture medium (indirect method) is as reliable for the diagnosis of bacterial keratitis as inoculation of multiple scrapes directly onto agar plates (direct method). To investigate this, bacterial recovery was assessed following transfer and transport of different concentrations and types of bacteria from an artificially contaminated surgical blade into brain heart infusion (BHI). Bacterial recovery rates between the proposed (indirect) and standard (direct) method were then compared after the in vitro inoculation of pig corneas and following specimen collection in patients with presumed bacterial ulcerative keratitis. Recovery of bacteria from contaminated surgical blades was found to be the same from both solid and liquid culture media. There was no significant difference in the numbers of positive cultures from solid (direct) and liquid (indirect) culture media, both in the experimental pig cornea inoculation study (P = 0.34) and in experiments with patients with clinical infections (P = 0.4), with an 85.2% agreement between methods (kappa = 0.61, P < 0.0001). In conclusion, therefore, the collection of two corneal scrapes, one used for Gram staining and the other transported in BHI followed by plating and subculturing in an enrichment medium, provides a simple method for the investigation of presumed bacterial keratitis. PMID:12843063

Hydrodynamics of bacterial colonies: Phase diagrams

NASA Astrophysics Data System (ADS)

Lega, J.; Passot, T.

2004-09-01

We present numerical simulations of a recent hydrodynamic model describing the growth of bacterial colonies on agar plates. We show that this model is able to qualitatively reproduce experimentally observed phase diagrams, which relate a colony shape to the initial quantity of nutrients on the plate and the initial wetness of the agar. We also discuss the principal features resulting from the interplay between hydrodynamic motions and colony growth, as described by our model.

MALDI-TOF mass spectrometry and high-consequence bacteria: safety and stability of biothreat bacterial sample testing in clinical diagnostic laboratories.

PubMed

Tracz, Dobryan M; Tober, Ashley D; Antonation, Kym S; Corbett, Cindi R

2018-03-01

We considered the application of MALDI-TOF mass spectrometry for BSL-3 bacterial diagnostics, with a focus on the biosafety of live-culture direct-colony testing and the stability of stored extracts. Biosafety level 2 (BSL-2) bacterial species were used as surrogates for BSL-3 high-consequence pathogens in all live-culture MALDI-TOF experiments. Viable BSL-2 bacteria were isolated from MALDI-TOF mass spectrometry target plates after 'direct-colony' and 'on-plate' extraction testing, suggesting that the matrix chemicals alone cannot be considered sufficient to inactivate bacterial culture and spores in all samples. Sampling of the instrument interior after direct-colony analysis did not recover viable organisms, suggesting that any potential risks to the laboratory technician are associated with preparation of the MALDI-TOF target plate before or after testing. Secondly, a long-term stability study (3 years) of stored MALDI-TOF extracts showed that match scores can decrease below the threshold for reliable species identification (<1.7), which has implications for proficiency test panel item storage and distribution.

Cutaneous antimicrobial preparation prior to intravenous catheterization in healthy dogs: clinical, microbiological, and histopathological evaluation.

PubMed

Coolman, B R; Marretta, S M; Kakoma, I; Wallig, M A; Coolman, S L; Paul, A J

1998-12-01

The purpose of this study was to determine the effects of a one-minute chlorhexidine gluconate skin preparation protocol prior to cephalic vein catheterization. Twenty-three healthy beagle dogs had one leg aseptically prepared and the opposite leg served as a control. Twenty-six- and 77-hour time groups were studied. Chlorhexidine-treated legs had significantly lower cutaneous bacterial counts than the control legs prior to catheter insertion and prior to catheter withdrawal for both time groups. Control legs developed significantly more dermatitis than the treated legs after 77 h. A one-minute preparation with 4% chlorhexidine gluconate was an effective method for sustained reduction of cutaneous bacterial counts at peripheral intravenous catheter insertion points in dogs. Increased cutaneous bacterial counts were associated with significantly more microscopic dermatitis in untreated legs after 77 h of catheterization.

Growth of methylaminotrophic, acetotrophic and hydrogenotrophic methanogenic bacteria on artificial supports.

PubMed

Urrutia, H; Vidal, R; Baeza, M; Reyes, J E; Aspe, E

1997-06-01

The efficiency of organic matter degradation in attached biomass reactors depends on the suitable selection of artificial support for the retention of bacterial communities. We have studied the growth on glass and clay beads of methylaminotrophic, acetotrophic and hydrogenotrophic methanogenic bacterial communities isolated from anaerobic reactors. Bacterial counts were performed by the standard MPN technique. Experiments were performed in 50 ml vials for 12 days at 35 degrees C. Increase in the counts of methylaminotrophic and hydrogenotrophic methanogens occurred on both glass and clay beads. The latter support material also stimulated the growth rate of methylaminotrophic methanogens.

Characterization of reticulated vitreous carbon foam using a frisch-grid parallel-plate ionization chamber

NASA Astrophysics Data System (ADS)

Edwards, Nathaniel S.; Conley, Jerrod C.; Reichenberger, Michael A.; Nelson, Kyle A.; Tiner, Christopher N.; Hinson, Niklas J.; Ugorowski, Philip B.; Fronk, Ryan G.; McGregor, Douglas S.

2018-06-01

The propagation of electrons through several linear pore densities of reticulated vitreous carbon (RVC) foam was studied using a Frisch-grid parallel-plate ionization chamber pressurized to 1 psig of P-10 proportional gas. The operating voltages of the electrodes contained within the Frisch-grid parallel-plate ionization chamber were defined by measuring counting curves using a collimated 241Am alpha-particle source with and without a Frisch grid. RVC foam samples with linear pore densities of 5, 10, 20, 30, 45, 80, and 100 pores per linear inch were separately positioned between the cathode and anode. Pulse-height spectra and count rates from a collimated 241Am alpha-particle source positioned between the cathode and each RVC foam sample were measured and compared to a measurement without an RVC foam sample. The Frisch grid was positioned in between the RVC foam sample and the anode. The measured pulse-height spectra were indiscernible from background and resulted in negligible net count rates for all RVC foam samples. The Frisch grid parallel-plate ionization chamber measurement results indicate that electrons do not traverse the bulk of RVC foam and consequently do not produce a pulse.

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.

Efficacy of High-volume Evacuator in Aerosol Reduction: Truth or Myth? A Clinical and Microbiological Study.

PubMed

Desarda, Hitesh; Gurav, Abhijit; Dharmadhikari, Chandrakant; Shete, Abhijeet; Gaikwad, Subodh

2014-01-01

Background and aims. Basic periodontal treatment aims at eliminating supra- and sub-gingival plaque and establishing conditions which will allow effective self-performed plaque control. This aim is primarily achieved with sonic and ultrasonic scalers. However, generation of bacterial aerosols during these procedures is of great concern to patients, the dentist and the dental assistant. The aim of this study was to compare the reduction in aerosol with and without high-volume evacuator through a microbiological study. Materials and methods. For this clinical study a fumigated closed operatory was selected. Maxillary incisors and canines were selected as an area for scaling. Piezoelectric ultrasonic scaling was performed in the absence and in the presence of a high-volume evacuator at 12 and 20 inches from the patient's oral cavity. In both groups scaling was carried out for 10 minutes. Nutrient agar plates were exposed for a total of 20 minutes. After this procedure, nutrient agar plates were incubated in an incubator at 37°C for 24 hours. The next day the nutrient agar plates were examined for colony forming units by a single microbiologist. Results. The results showed no statistically significant differences in colony forming units (CFU) with and without the use of a high-volume evacuator either at 12 or 20 inches from the patient's oral cavity. Conclusion. It was concluded that high-volume evacuator, when used as a separate unit without any modification, is not effective in reducing aerosol counts and environmental contamination.

[Incidence of Campylobacter spp. and Salmonella spp. in raw and roasted chicken in Guadalajara, Mexico].

PubMed

Castillo-Ayala, A; Salas-Ubiarco, M G; Márquez-Padilla, M L; Osorio-Hernández, M D

1993-01-01

The presence of Campylobacter spp. and Salmonella was studied in 70 samples of fresh retail chicken pieces and in 40 samples of roast chicken. Total plate count was performed in every sample as well. Most of the samples of fresh chicken yielded total plate counts > 10(8)/piece (thigh), while in roast chicken these counts ranged from 10(3) to 10(5)/piece (leg and thigh). Campylobacter was isolated from 33% of fresh chicken and from no sample of roast chicken. Salmonella was isolated from 69% of fresh chicken and 2.5% of roast chicken. There was no relationship between total plate counts in fresh chicken and isolation of either Campylobacter or Salmonella. Sixty percent of the Salmonella isolates belonged to serotype S. anatum, and about 50% of the isolates of Campylobacter were identified as being C. coli. The only Salmonella-positive sample of roast chicken yielded three serotypes: S. give, S. muenster, and S. manhattan. Presence of Campylobacter and Salmonella in chicken is of concern, due to the risk of spreading from the raw food to other cooked foods. The isolation of pathogens from roast chicken indicates mishandling during processing and/or storage of the product.

A generalized plate method for estimating total aerobic microbial count.

PubMed

Ho, Kai Fai

2004-01-01

The plate method outlined in Chapter 61: Microbial Limit Tests of the U.S. Pharmacopeia (USP 61) provides very specific guidance for assessing total aerobic bioburden in pharmaceutical articles. This methodology, while comprehensive, lacks the flexibility to be useful in all situations. By studying the plate method as a special case within a more general family of assays, the effects of each parameter in the guidance can be understood. Using a mathematical model to describe the plate counting procedure, a statistical framework for making more definitive statements about total aerobic bioburden is developed. Such a framework allows the laboratory scientist to adjust the USP 61 methods to satisfy specific practical constraints. In particular, it is shown that the plate method can be conducted, albeit with stricter acceptance criteria, using a test specimen quantity that is smaller than the 10 g or 10 mL prescribed in the guidance. Finally, the interpretation of results proffered by the guidance is re-examined within this statistical framework and shown to be overly aggressive.

The Accuracy of the Sysmex UF-1000i in Urine Bacterial Detection Compared With the Standard Urine Analysis and Culture.

PubMed

Erdman, Patrick; Anderson, Brian; Zacko, J Christopher; Taylor, Kirk; Donaldson, Keri

2017-11-01

- Urinary tract infections are characterized by the presence of microbial pathogens within the urinary tract. They represent one of the most common infections in hospitalized and clinic patients. - To model the parameters of the Sysmex UF-1000i to the gold standard, urine culture, and to compare the detection of dipstick leukocyte esterase and nitrates to urine cultures and UF-1000i results. - Data were compared from urine samples collected in sterile containers for bacterial culture and microscopic analysis. One sample was used to inoculate a 5% sheep blood agar and MacConkey agar plate using a 0.001-mL calibrated loop. The second sample was analyzed by urinalysis-associated microscopy. The media plates were investigated for growth after 18 to 24 hours of aerobic incubation at 37°C. The second sample was analyzed for bacteria and leukocytes with the Sysmex UF-1000i according to the manufacturer's guidelines. Three definitions for culture results, sensitivity, and specificity at different cutoff values were calculated for the UF-1000i. - The negative predictive value for any positive culture in the adult population included in the study was 95.5%, and the negative predictive value for positive cultures containing growth of 100 000 or more colony-forming units was 99.3% using the Sysmex UF-1000i. - Sysmex UF-1000i showed 98% sensitivity and 93.7% specificity with a 95.5% negative predictive value. Thus, a negative screen with the UF-1000i using defined thresholds for white blood cell counts and bacteria was likely to be a true negative, decreasing the need for presumptive antibiotics.

Long-term clinical and bacterial effects of xylitol on patients with fixed orthodontic appliances.

PubMed

Masoud, Mohamed I; Allarakia, Reem; Alamoudi, Najlaa M; Nalliah, Romesh; Allareddy, Veerasathpurush

2015-01-01

The objective of this study was to evaluate long-term clinical and bacterial effects of using 6 g of xylitol per day for 3 months on patients with full fixed orthodontic appliances. The study was a pilot clinical trial that included 41 subjects who were undergoing orthodontic treatment. The subjects were randomly divided into three groups. Group A received xylitol chewing gum, group B received xylitol dissolvable chewable tablets, and Group C served as the control group and did not receive xylitol gums or tablets. Clinical examination and the collection of plaque and saliva samples were carried out at baseline and 3, 6, and 12 months. All three groups were given oral hygiene instruction and were put on a 6-month cleaning and topical fluoride schedule. Plaque scores and bacterial counts were used to evaluate the effectiveness of the different approaches at reducing the caries risk. Xylitol groups did not experience any more reduction in plaque score, plaque MS counts, or salivary MS counts than the control group nor did they have lower values at any of the time points. Chewing gum did not significantly increase the incidence of debonded brackets over the other groups. Xylitol does not have a clinical or bacterial benefit in patients with fixed orthodontic appliances. Oral hygiene instructions and 6-month topical fluoride application were effective at reducing plaque scores and bacterial counts in patients with full fixed appliances regardless of whether or not xylitol was used.

Analysis of clinical outcomes in pediatric bacterial meningitis focusing on patients without cerebrospinal fluid pleocytosis.

PubMed

Lin, Wen-Li; Chi, Hsin; Huang, Fu-Yuan; Huang, Daniel Tsung-Ning; Chiu, Nan-Chang

2016-10-01

Cerebrospinal fluid (CSF) cell count and biochemical examinations and cultures form the basis for the diagnosis of bacterial meningitis. However, some patients do not have typical findings and are at a higher risk of being missed or having delayed treatment. To better understand the correlation between CSF results and outcomes, we evaluated CSF data focusing on the patients with atypical findings. This study enrolled CSF culture-proven bacterial meningitis patients aged from 1 month to 18 years in a medical center. The patients were divided into "normal" and "abnormal" groups for each laboratory result and in combination. The correlations between the laboratory results and the outcomes were analyzed. A total of 175 children with confirmed bacterial meningitis were enrolled. In CSF examinations, 16.2% of patients had normal white blood cell counts, 29.5% had normal glucose levels, 24.5% had normal protein levels, 10.2% had normal results in two items, and 8.6% had normal results in all three items. In logistic regression analysis, a normal CSF leukocyte count and increased CSF protein level were related to poor outcomes. Patients with meningitis caused by Streptococcus pneumoniae and hyponatremia were at a higher risk of mortality and the development of sequelae. In children with bacterial meningitis, nontypical CSF findings and, in particular, normal CSF leukocyte count and increased protein level may indicate a worse prognosis. Copyright © 2014. Published by Elsevier B.V.

Development of a Flow Cytometry-Based Method for Rapid Detection of Escherichia coli and Shigella Spp. Using an Oligonucleotide Probe

PubMed Central

Xue, Yong; Wilkes, Jon G.; Moskal, Ted J.; Williams, Anna J.; Cooper, Willie M.; Nayak, Rajesh; Rafii, Fatemeh; Buzatu, Dan A.

2016-01-01

Standard methods to detect Escherichia coli contamination in food use the polymerase chain reaction (PCR) and agar culture plates. These methods require multiple incubation steps and take a long time to results. An improved rapid flow-cytometry based detection method was developed, using a fluorescence-labeled oligonucleotide probe specifically binding a16S rRNA sequence. The method positively detected 51 E. coli isolates as well as 4 Shigella species. All 27 non-E. coli strains tested gave negative results. Comparison of the new genetic assay with a total plate count (TPC) assay and agar plate counting indicated similar sensitivity, agreement between cytometry cell and colony counts. This method can detect a small number of E.coli cells in the presence of large numbers of other bacteria. This method can be used for rapid, economical, and stable detection of E. coli and Shigella contamination in the food industry and other contexts. PMID:26913737

Development of a Flow Cytometry-Based Method for Rapid Detection of Escherichia coli and Shigella Spp. Using an Oligonucleotide Probe.

PubMed

Xue, Yong; Wilkes, Jon G; Moskal, Ted J; Williams, Anna J; Cooper, Willie M; Nayak, Rajesh; Rafii, Fatemeh; Buzatu, Dan A

2016-01-01

Standard methods to detect Escherichia coli contamination in food use the polymerase chain reaction (PCR) and agar culture plates. These methods require multiple incubation steps and take a long time to results. An improved rapid flow-cytometry based detection method was developed, using a fluorescence-labeled oligonucleotide probe specifically binding a16S rRNA sequence. The method positively detected 51 E. coli isolates as well as 4 Shigella species. All 27 non-E. coli strains tested gave negative results. Comparison of the new genetic assay with a total plate count (TPC) assay and agar plate counting indicated similar sensitivity, agreement between cytometry cell and colony counts. This method can detect a small number of E.coli cells in the presence of large numbers of other bacteria. This method can be used for rapid, economical, and stable detection of E. coli and Shigella contamination in the food industry and other contexts.

Method of detecting and counting bacteria in body fluids

NASA Technical Reports Server (NTRS)

Chappelle, E. W.; Picciolo, G. L. (Inventor)

1973-01-01

A novel method is reported for determining bacterial levels in urine samples, which method depends on the quantitative determination of bacterial adenosine triphosphate (ATP) in the presence of non-bacterial ATP. After the removal of non-bacterial ATP, the bacterial ATP is released by cell rupture and is measured by an enzymatic bioluminescent assay using an enzyme obtained from the firefly.

Differential Attachment of Salmonella enterica and Enterohemorrhagic Escherichia coli to Alfalfa, Fenugreek, Lettuce, and Tomato Seeds

PubMed Central

Cui, Yue; Walcott, Ronald

2017-01-01

ABSTRACT Vegetable seeds have the potential to disseminate and transmit foodborne bacterial pathogens. This study was undertaken to assess the abilities of selected Salmonella and enterohemorrhagic Escherichia coli (EHEC) strains to attach to fungicide-treated versus untreated, and intact versus mechanically damaged, seeds of alfalfa, fenugreek, lettuce, and tomato. Surface-sanitized seeds (2 g) were exposed to four individual strains of Salmonella or EHEC at 20°C for 5 h. Contaminated seeds were rinsed twice, each with 10 ml of sterilized water, before being soaked overnight in 5 ml of phosphate-buffered saline at 4°C. The seeds were then vortexed vigorously for 1 min, and pathogen populations in seed rinse water and soaking buffer were determined using a standard plate count assay. In general, the Salmonella cells had higher attachment ratios than the EHEC cells. Lettuce seeds by unit weight had the highest numbers of attached Salmonella or EHEC cells, followed by tomato, alfalfa, and fenugreek seeds. In contrast, individual fenugreek seeds had more attached pathogen cells, followed by lettuce, alfalfa, and tomato seeds. Significantly more Salmonella and EHEC cells attached to mechanically damaged seeds than to intact seeds (P < 0.05). Although, on average, significantly more Salmonella and EHEC cells were recovered from untreated than fungicide-treated seeds (P < 0.05), fungicide treatment did not significantly affect the attachment of individual bacterial strains to vegetable seeds (P > 0.05), with a few exceptions. This study fills gaps in the current body of literature and helps explain bacterial interactions with vegetable seeds with differing surface characteristics. IMPORTANCE Vegetable seeds, specifically sprout seeds, have the potential to disseminate and transmit foodborne bacterial pathogens. This study investigated the interaction between two important bacterial pathogens, i.e., Salmonella and EHEC, and vegetable seeds with differing surface characteristics. This research helps understand whether seed surface structure, integrity, and fungicide treatment affect the interaction between bacterial cells and vegetable seeds. PMID:28130295

Differential Attachment of Salmonella enterica and Enterohemorrhagic Escherichia coli to Alfalfa, Fenugreek, Lettuce, and Tomato Seeds.

PubMed

Cui, Yue; Walcott, Ronald; Chen, Jinru

2017-04-01

Vegetable seeds have the potential to disseminate and transmit foodborne bacterial pathogens. This study was undertaken to assess the abilities of selected Salmonella and enterohemorrhagic Escherichia coli (EHEC) strains to attach to fungicide-treated versus untreated, and intact versus mechanically damaged, seeds of alfalfa, fenugreek, lettuce, and tomato. Surface-sanitized seeds (2 g) were exposed to four individual strains of Salmonella or EHEC at 20°C for 5 h. Contaminated seeds were rinsed twice, each with 10 ml of sterilized water, before being soaked overnight in 5 ml of phosphate-buffered saline at 4°C. The seeds were then vortexed vigorously for 1 min, and pathogen populations in seed rinse water and soaking buffer were determined using a standard plate count assay. In general, the Salmonella cells had higher attachment ratios than the EHEC cells. Lettuce seeds by unit weight had the highest numbers of attached Salmonella or EHEC cells, followed by tomato, alfalfa, and fenugreek seeds. In contrast, individual fenugreek seeds had more attached pathogen cells, followed by lettuce, alfalfa, and tomato seeds. Significantly more Salmonella and EHEC cells attached to mechanically damaged seeds than to intact seeds ( P < 0.05). Although, on average, significantly more Salmonella and EHEC cells were recovered from untreated than fungicide-treated seeds ( P < 0.05), fungicide treatment did not significantly affect the attachment of individual bacterial strains to vegetable seeds ( P > 0.05), with a few exceptions. This study fills gaps in the current body of literature and helps explain bacterial interactions with vegetable seeds with differing surface characteristics. IMPORTANCE Vegetable seeds, specifically sprout seeds, have the potential to disseminate and transmit foodborne bacterial pathogens. This study investigated the interaction between two important bacterial pathogens, i.e., Salmonella and EHEC, and vegetable seeds with differing surface characteristics. This research helps understand whether seed surface structure, integrity, and fungicide treatment affect the interaction between bacterial cells and vegetable seeds. Copyright © 2017 American Society for Microbiology.

High temperature, short time pasteurization temperatures inversely affect bacterial numbers during refrigerated storage of pasteurized fluid milk.

PubMed

Ranieri, M L; Huck, J R; Sonnen, M; Barbano, D M; Boor, K J

2009-10-01

The grade A Pasteurized Milk Ordinance specifies minimum processing conditions of 72 degrees C for at least 15 s for high temperature, short time (HTST) pasteurized milk products. Currently, many US milk-processing plants exceed these minimum requirements for fluid milk products. To test the effect of pasteurization temperatures on bacterial numbers in HTST pasteurized milk, 2% fat raw milk was heated to 60 degrees C, homogenized, and treated for 25 s at 1 of 4 different temperatures (72.9, 77.2, 79.9, or 85.2 degrees C) and then held at 6 degrees C for 21 d. Aerobic plate counts were monitored in pasteurized milk samples at d 1, 7, 14, and 21 postprocessing. Bacterial numbers in milk processed at 72.9 degrees C were lower than in milk processed at 85.2 degrees C on each sampling day, indicating that HTST fluid milk-processing temperatures significantly affected bacterial numbers in fluid milk. To assess the microbial ecology of the different milk samples during refrigerated storage, a total of 490 psychrotolerant endospore-forming bacteria were identified using DNA sequence-based subtyping methods. Regardless of processing temperature, >85% of the isolates characterized at d 0, 1, and 7 postprocessing were of the genus Bacillus, whereas more than 92% of isolates characterized at d 14 and 21 postprocessing were of the genus Paenibacillus, indicating that the predominant genera present in HTST-processed milk shifted from Bacillus spp. to Paenibacillus spp. during refrigerated storage. In summary, 1) HTST processing temperatures affected bacterial numbers in refrigerated milk, with higher bacterial numbers in milk processed at higher temperatures; 2) no significant association was observed between genus isolated and pasteurization temperature, suggesting that the genera were not differentially affected by the different processing temperatures; and 3) although typically present at low numbers in raw milk, Paenibacillus spp. are capable of growing to numbers that can exceed Pasteurized Milk Ordinance limits in pasteurized, refrigerated milk.

Evaluation of the antibacterial effects of vancomycin hydrochloride released from agar-gelatin-bioactive glass composites.

PubMed

Rivadeneira, Josefina; Di Virgilio, Ana Laura; Audisio, M Carina; Boccaccini, Aldo R; Gorustovich, Alejandro A

2015-01-13

The aim of this work was to evaluate the perfomance of agar-gelatin (AG) composites and AG-containing 45S5 bioactive glass (BG) microparticles (AGBG) in relation to their water uptake capacity, sustained release of a drug over time, and antibacterial effects. The composites were fabricated by the gel-casting method. To impart the local drug release capacity, vancomycin hydrochloride (VC) was loaded in the composites in concentrations of 0.5 and 1 mg ml(-1). VC release was assessed in distilled water at 37 °C up to 72 h and quantified spectrophotometrically. The antibacterial activity of composites was evaluated by the inhibition zone test and the plate count method. The experiments were performed in vitro up to 48 h on three staphylococcus strains: Staphylococcus aureus ATCC29213, S. aureus ATCC6538 and Staphylococcus epidermidis ATCC12228. The results showed that the addition of BG to AG composites did not affect the degree of water uptake. The release of VC was significantly affected by the presence of BG. VC release was higher from AGBGVC films than from AGVC ones over prolonged incubation times. Bacterial inhibition zones were found around the composites. The halos were larger when the cells were put in contact with AGVC composites than when they were put in contact with AGBGVC ones. Nevertheless, the viable count method demonstrated that the composites inhibited Staphylococcus cell growth with no statistical differences. In conclusion, the addition of BG did not reflect an improvement in the parameters studied. On the other hand, composites loaded with VC would have a role in prophylaxis against bacterial infection.

The effect of water extract of sumac (Rhus coriaria L.) and lactic acid on decontamination and shelf life of raw broiler wings.

PubMed

Gulmez, M; Oral, N; Vatansever, L

2006-08-01

In an attempt to improve the bacteriological quality and refrigerated shelf life of broiler meat, 10-min surface wash treatments with sterile distilled water (DW), 8% (wt/vol) water extract of sumac (Rhus coriaria L.) fruits (WES), and 2% (vol/vol) lactic acid (LA) were compared by using a broiler wing model. The aerobic plate counts (log10 cfu/g) of psychrotrophs, mesophilic aerobes, Enterobacteriaceae, coliforms and presumptive fecal coliforms on the samples were determined. Immediately after a 10-min decontaminaton, the mean count of all the bacterial groups was determined to be 3.9, 2.6, and 1.7 (log10 cfu/g) for DW, WES, and LA, respectively. Because the postdecontamination population level of psychrotrophs, mesophiles, and Enterobacteriaceae were low in the LA-treated group compared to the WES group, an equity between the 2 groups in the point of view of the 3 bacterial groups existed at d 10 of cold storage (3 +/- 1 degrees C). Shelf life was 7 and 14 d for wings treated with DW and WES, respectively, whereas the LA-treated wings did not spoil after 14 d of cold storage (3 +/- 1 degrees C). Nevertheless, an undesirable pale color and an acidulous odor occurred in the LA-treated wings. In contrast, a good color appeared on the WES-treated wings, which was also superior to the color of the DW-treated wings. Such advantages of WES may be important for poultry processors and for consumers. However, the immediate decontamination and refrigerated shelf life extension potential of WES should be intensively studied in antimicrobial interventions in poultry processing plants.

Effect of refrigeration and frozen storage on the Campylobacter jejuni recovery from naturally contaminated broiler carcasses

PubMed Central

Maziero, Maike T.; de Oliveira, Tereza Cristina R. M.

2010-01-01

Campylobacter jejuni is the most common thermophilic Campylobacter associated with human enteritis in many countries. Broilers and their by-products are the main sources for human enteritis. Refrigeration and freezing are used to control bacterial growth in foods. The effect of these interventions on survival of Campylobacter jejuni is yet not quite understood. This study evaluated the effect of storage temperature on the survival of C. jejuni in chicken meat stored for seven days at 4°C and for 28 days at -20°C. The influence of selective enrichment on recovery of Campylobacter was also evaluated. Thirty fresh chicken meat samples were analyzed and 93.3% was contaminated with termotolerant Campylobacter spp. with average count of 3.08 Log10 CFU/g on direct plating. After refrigeration, 53.3% of the analyzed samples tested positive for Campylobacter and the average count was 1.19 Log10 CFU/g. After storage at -20°C, 36.6% of the samples were positive with a verage count of 0.75 Log10 CFU/g. C. jejuni was detected after enrichment, respectively, in 50% of the fresh, 36.7% of the refrigerated and 33.3% of the frozen meat samples analyzed. No difference was detected for the recovery of C. jejuni from fresh, refrigerated or frozen samples after selective enrichment, showing that this microorganism can survive under the tested storage conditions. PMID:24031523

Bacterial Quality and Prevalence of Foodborne Pathogens in Edible Offal from Slaughterhouses in Korea.

PubMed

Im, Min Chan; Seo, Kwang Won; Bae, Dong Hwa; Lee, Young Ju

2016-01-01

Edible offal meats have recently received significant attention worldwide. However, studies evaluating the microbial quality of diverse edible offal and specifically investigating contamination by pathogens that cause foodborne illnesses are rare. Our study was conducted to investigate the microbiological quality of six kinds of edible offal produced from 11 pigs and 8 cattle slaughterhouses in the Republic of Korea and the prevalence of pathogenic microorganisms such as Salmonella, Clostridium perfringens, Staphylococcus aureus, and Escherichia coli O157:H7 in these products. The values for aerobic plate counts, coliform counts, and E. coli counts in red offal were 1.00 to 6.70, 0 (below 10 CFU) to 4.78, and 0 to 4.00 log CFU/g, respectively. For green offal, the values were 3.00 to 7.00, 1.48 to 6.30, and 0 to 6.00 log CFU/g, respectively. The most frequently detected foodborne pathogen was Salmonella (23.8% prevalence in pig offal and 7.1% prevalence in cattle offal), followed by C. perfringens (11.1 and 7.1%, respectively) and S. aureus (12.7 and 2.4%, respectively). None of the offal samples tested positive for E. coli O157:H7. Considering the microbial quality of offal from Korean slaughterhouses and the prevalence of foodborne pathogens in this material, more refined hygienic standards such as a hazard analysis critical control point system for processing, packing, and transporting edible offal are necessary for preventing further contamination.

Microbiologic results after non-surgical erbium-doped:yttrium, aluminum, and garnet laser or air-abrasive treatment of peri-implantitis: a randomized clinical trial.

PubMed

Persson, G Rutger; Roos-Jansåker, Ann-Marie; Lindahl, Christel; Renvert, Stefan

2011-09-01

The purpose of this study is to assess clinical and microbiologic effects of the non-surgical treatment of peri-implantitis lesions using either an erbium-doped:yttrium, aluminum, and garnet (Er:YAG) laser or an air-abrasive subgingival polishing method. In a 6-month clinical trial, 42 patients with peri-implantitis were treated at one time with an Er:YAG laser or an air-abrasive device. Routine clinical methods were used to monitor clinical conditions. Baseline and 6-month intraoral radiographs were assessed with a software program. The checkerboard DNA-DNA hybridization method was used to assess 74 bacterial species from the site with the deepest probing depth (PD) at the implant. Non-parametric tests were applied to microbiology data. PD reductions (mean ± SD) were 0.9 ± 0.8 mm and 0.8 ± 0.5 mm in the laser and air-abrasive groups, respectively (not significant). No baseline differences in bacterial counts between groups were found. In the air-abrasive group, Pseudomonas aeruginosa, Staphylococcus aureus, and Staphylococcus anaerobius were found at lower counts at 1 month after therapy (P <0.001) and with lower counts in the laser group for Fusobacterium nucleatum naviforme (P = 0.002), and Fusobacterium nucleatum nucleatum (P = 0.002). Both treatments failed to reduce bacterial counts at 6 months. Porphyromonas gingivalis counts were higher in cases with progressive peri-implantitis (P <0.001). At 1 month, P. aeruginosa, S. aureus, and S. anaerobius were reduced in the air-abrasive group, and Fusobacterium spp. were reduced in the laser group. Six-month data demonstrated that both methods failed to reduce bacterial counts. Clinical improvements were limited.

Development of a rapid optic bacteria detecting system based on ATP bioluminescence

NASA Astrophysics Data System (ADS)

Liu, Jun Tao; Luo, JinPing; Liu, XiaoHong; Cai, XinXia

2014-12-01

A rapid optic bacteria detecting system based on the principle of Adenosine triphosphate(ATP) bioluminescence was presented in this paper. This system consisted of bioluminescence-based biosensor and the high-sensitivity optic meter. A photon counting photomultiplier tube (PMT) module was used to improve the detection sensitivity, and a NIOS II/f processor based on a Field Programmable Gate Array(FPGA) was used to control the system. In this work, Micrococcus luteus were chosen as the test sample. Several Micrococcus luteus suspension with different concentration was tested by both T2011 and plate counting method. By comparing the two group results, an calibration curve was obtained from the bioluminescence intensity for Micrococcus luteus in the range of 2.3×102 ~ 2.3×106 CFU/mL with a good correlation coefficient of 0.960. An impacting Air microorganism sampler was used to capture Airborne Bacteria, and 8 samples were collected in different place. The TBC results of 8 samples by T2011 were between 10 ~ 2×103 cfu/mL, consistent with that of plate counting method, which indicated that 8 samples were between 10 ~ 3×103 cfu/mL. For total airborne bacteria count was small, correlation coefficient was poor. Also no significant difference was found between T2011 and plate counting method by statistical analyses.

Rapid enumeration of viable bacteria by image analysis

NASA Technical Reports Server (NTRS)

Singh, A.; Pyle, B. H.; McFeters, G. A.

1989-01-01

A direct viable counting method for enumerating viable bacteria was modified and made compatible with image analysis. A comparison was made between viable cell counts determined by the spread plate method and direct viable counts obtained using epifluorescence microscopy either manually or by automatic image analysis. Cultures of Escherichia coli, Salmonella typhimurium, Vibrio cholerae, Yersinia enterocolitica and Pseudomonas aeruginosa were incubated at 35 degrees C in a dilute nutrient medium containing nalidixic acid. Filtered samples were stained for epifluorescence microscopy and analysed manually as well as by image analysis. Cells enlarged after incubation were considered viable. The viable cell counts determined using image analysis were higher than those obtained by either the direct manual count of viable cells or spread plate methods. The volume of sample filtered or the number of cells in the original sample did not influence the efficiency of the method. However, the optimal concentration of nalidixic acid (2.5-20 micrograms ml-1) and length of incubation (4-8 h) varied with the culture tested. The results of this study showed that under optimal conditions, the modification of the direct viable count method in combination with image analysis microscopy provided an efficient and quantitative technique for counting viable bacteria in a short time.

Low noise and conductively cooled microchannel plates

NASA Technical Reports Server (NTRS)

Feller, W. B.

1990-01-01

Microchannel plate (MCP) dynamic range has recently been enhanced for both very low and very high input flux conditions. Improvements in MCP manufacturing technology reported earlier have led to MCPs with substantially reduced radioisotope levels, giving dramatically lower internal background-counting rates. An update is given on the Galileo low noise MCP. Also, new results in increasing the MCP linear counting range for high input flux densities are presented. By bonding the active face of a very low resistance MCP (less than 1 megaohm) to a substrate providing a conductive path for heat transport, the bias current limit (hence, MCP output count rate limit) can be increased up to two orders of magnitude. Normal pulse-counting MCP operation was observed at bias currents of several mA when a curved-channel MCP (80:1) was bonded to a ceramic multianode substrate; the MCP temperature rise above ambient was less than 40 C.

Uncertainty of quantitative microbiological methods of pharmaceutical analysis.

PubMed

Gunar, O V; Sakhno, N G

2015-12-30

The total uncertainty of quantitative microbiological methods, used in pharmaceutical analysis, consists of several components. The analysis of the most important sources of the quantitative microbiological methods variability demonstrated no effect of culture media and plate-count techniques in the estimation of microbial count while the highly significant effect of other factors (type of microorganism, pharmaceutical product and individual reading and interpreting errors) was established. The most appropriate method of statistical analysis of such data was ANOVA which enabled not only the effect of individual factors to be estimated but also their interactions. Considering all the elements of uncertainty and combining them mathematically the combined relative uncertainty of the test results was estimated both for method of quantitative examination of non-sterile pharmaceuticals and microbial count technique without any product. These data did not exceed 35%, appropriated for a traditional plate count methods. Copyright © 2015 Elsevier B.V. All rights reserved.

Photon-counting detector arrays based on microchannel array plates. [for image enhancement

NASA Technical Reports Server (NTRS)

Timothy, J. G.

1975-01-01

The recent development of the channel electron multiplier (CEM) and its miniaturization into the microchannel array plate (MCP) offers the possibility of fully combining the advantages of the photographic and photoelectric detection systems. The MCP has an image-intensifying capability and the potential of being developed to yield signal outputs superior to those of conventional photomultipliers. In particular, the MCP has a photon-counting capability with a negligible dark-count rate. Furthermore, the MCP can operate stably and efficiently at extreme-ultraviolet and soft X-ray wavelengths in a windowless configuration or can be integrated with a photo-cathode in a sealed tube for use at ultraviolet and visible wavelengths. The operation of one- and two-dimensional photon-counting detector arrays based on the MCP at extreme-ultraviolet wavelengths is described, and the design of sealed arrays for use at ultraviolet and visible wavelengths is briefly discussed.

Evaluation of commercially prepared transport systems for nonlethal detection of Aeromonas salmonicida in salmonid fish

USGS Publications Warehouse

Cipriano, R.C.; Bullock, G.L.

2001-01-01

In vitro studies indicated that commercially prepared transport systems containing Amies, Stuart's, and Cary-Blair media worked equally well in sustaining the viability of the fish pathogen Aeromonas salmonicida, which causes furunculosis. The bacterium remained viable without significant increase or decrease in cell numbers for as long as 48 h of incubation at 18-20??C in Stuart's transport medium; consequently, obtaining mucus samples in such tubes were comparable to on-site detection of A. salmonicida by dilution plate counts on Coomassie Brilliant Blue agar. In three different assays of 100 samples of mucus from Atlantic salmon Salmo salar infected subclinically with A. salmonicida, dilution counts conducted on-site proved more reliable for detecting the pathogen than obtaining the samples in the transport system. In the on-site assays, dilution counts detected the pathogen in 34, 41, and 22 samples, whereas this was accomplished in only 15, 15, and 3 of the respective samples when the transport system was used. In an additional experiment, Arctic char Salvelinus alpinus sustaining a frank epizootic of furunculosis were sampled similarly. Here, too, dilution counts were more predictive of the prevalence of A. salmonicida and detected the pathogen in 46 mucus samples; in comparison, only 6 samples collected by using the transport system were positive. We also observed that the transport system supported the growth of the normal mucus bacterial flora. Particularly predominant among these were motile aeromonads and Pseudomonas fluorescens. In studies of mixed culture growth, two representatives of both of the latter genera of bacteria outgrew A. salmonicida - in some cases, to the total exclusion of the pathogen itself.

Effects of bacterial pollution caused by a strong typhoon event and the restoration of a recreational beach: Transitions of fecal bacterial counts and bacterial flora in beach sand.

PubMed

Suzuki, Yoshihiro; Teranishi, Kotaro; Matsuwaki, Tomonori; Nukazawa, Kei; Ogura, Yoshitoshi

2018-05-28

To determine the effects of bacteria pollution associated with a strong typhoon event and to assess the restoration of the normal bacterial flora, we used conventional filtration methods and nextgeneration sequencing of 16S rRNA genes to analyze the transition of fecal and total bacterial counts in water and core sand samples collected from a recreational beach. Immediately after the typhoon event, Escherichia coli counts increased to 82 CFU/100 g in the surface beach sand. E. coli was detected through the surface to sand 85-cm deep at the land side point (10-m land side from the high-water line). However, E. coli disappeared within a month from the land side point. The composition of the bacterial flora in the beach sand at the land point was directly influenced by the typhoon event. Pseudomonas was the most prevalent genus throughout the sand layers (0-102-cm deep) during the typhoon event. After 3 months, the population of Pseudomonas significantly decreased, and the predominant genus in the surface layer was Kaistobacter, although Pseudomonas was the major genus in the 17- to 85-cm layer. When the beach conditions stabilized, the number of pollutant Pseudomonas among the 10 most abundant genera decreased to lower than the limit of detection. The bacterial population of the sand was subsequently restored to the most populous pre-event orders at the land point. A land-side beach, where users directly contact the sand, was significantly affected by bacterial pollution caused by a strong typhoon event. We show here that the normal bacterial flora of the surface sand was restored within 1 month. Copyright © 2018 Elsevier B.V. All rights reserved.

The development of a new three-step protocol to determine the efficacy of disinfectant wipes on surfaces contaminated with Staphylococcus aureus.

PubMed

Williams, G J; Denyer, S P; Hosein, I K; Hill, D W; Maillard, J-Y

2007-12-01

We developed a three-step protocol to quantify the efficacy of disinfectant wipes, their ability to remove and prevent microbial transfer from surfaces and their overall antimicrobial activity. Meticillin-resistant (MRSA) or -susceptible (MSSA) Staphylococcus aureus (6-7 log(10)cfu) were inoculated onto stainless steel discs with or without organic load and dried. Grapefruit extract-containing test wipes and unmedicated control wipes were used. In step 1, wipes were mechanically rotated against surfaces for 10s at 60rpm, exerting a weight of 100+/-5g. Bacterial removal was assessed by transferring the steel discs to neutraliser, resuspending and counting remaining bacteria. In step 2, bacterial transfer from wipes was assessed by eight consecutive mechanical adpression transfers to agar/neutraliser plates. Step 3 was the measurement of antimicrobial activity by direct inoculation of the wipes for 10s followed by neutralisation and enumeration. Test wipes achieved a significantly higher bacterial cell removal than control wipes on all surfaces (P<0.05). The low bactericidal activity of the wipes (<1 log(10) reduction when directly inoculated) and the subsequent survival of bacteria on the wipes, however, led to repeated microbial transfer when initially high contamination levels were present. There were no differences between MRSA and MSSA in removal, transfer or antimicrobial activity. The three-step method is a useful tool for developing future guidelines to assess the ability of wipes to disinfect surfaces.

Magazines in waiting areas of hospital: a forgotten microbial reservoir?

PubMed

Adé, Mathias; Burger, Sandrine; Cuntzmann, Anaelle; Exinger, Julien; Meunier, Olivier

2017-12-01

The hospital environment is a potential source of microbial contamination. Thus, the magazines in hospital's waiting rooms are handled by patients and visitors whose health and hygiene conditions can vary widely. In this context, we had measured the microbial load on the surface of magazines. Fifteen magazines from 5 waiting rooms of hospital are sampled by agar prints at the areas taken in hand. The agar plates are incubated at 30̊C for 72h. The colonies are counted and identified by MALDI-TOF mass spectrometry (Vitek ® -MS). The extraction efficiency of bacteria by the agar print method on the magazines is calculated. All the samples highlight a varied bacterial flora: 32CFU/agar in mean. Isolated bacteria come principally from the skin flora (>60%), but we also isolate potentially pathogenic micro-organisme like S. aureus, E. faecalis, A. viridans and Aspergillus sp. as well as oropharyngeal flora bacteria like A. iwolfii and M. osloensis and fecal like B. stercoris. Some species rarely described in hospital are also isolated such as P. yeei or K. sedentarius. The extraction efficiency of the sampling method on a magazine is 36%. Our study, which is the first to be interested in the bacterial contamination of magazines in hospital, could make them consider as microbial reservoir to be controlled, especially for the most fragile patients. New bacterial identification techniques as the MALDI-TOF allow to reveal the presence of rarely described and often underestimated species.

Microbial diversity of consumption milk during processing and storage.

PubMed

Porcellato, Davide; Aspholm, Marina; Skeie, Siv Borghild; Monshaugen, Marte; Brendehaug, Johanne; Mellegård, Hilde

2018-02-02

Bovine milk contains a complex microbial community that affects the quality and safety of the product. Detailed knowledge of this microbiota is, therefore, of importance for the dairy industry. In this study, the bacterial composition of consumption milk was assessed during different stages in the production line and throughout the storage in cartons by using culturing techniques and 16S rRNA marker gene sequencing. Monthly samples from two dairies were analyzed to capture the seasonal variations in the milk microbiota. Although there was a core microbiota present in milk samples from both dairies, the composition of the bacterial communities were significantly influenced by sampling month, processing stage and storage temperature. Overall, a higher abundance of operational taxonomic units (OTUs) within the order Bacillales was detected in samples of raw and pasteurized milk from the spring and summer months, while Pseudomonadales and Lactobacillales OTUs were predominant in the winter months. OTUs belonging to the order Lactobacillales, Pseudomonadales, Clostridiales and Bacillales were significantly more abundant in milk samples taken immediately after pasteurization compared to raw milk samples. During storage of milk in cartons at 4°C, the bacterial composition remained stable throughout the product shelf life, while storage at 8°C significantly increased the abundance of OTUs belonging to the genus Bacillus and the plate count levels of presumptive Bacillus cereus. The knowledge obtained in this work will be useful to the dairy industry during their quality assurance work and risk assessment practices. Copyright © 2017 Elsevier B.V. All rights reserved.

The effect of a honey based gel and silver sulphadiazine on bacterial infections of in vitro burn wounds.

PubMed

Boekema, B K H L; Pool, L; Ulrich, M M W

2013-06-01

Bacterial contamination remains a constant threat in burn wound care. Topical treatments to combat contaminations have good bactericidal effects but can have detrimental effects for the healing process. Treatments with for example silver can increase healing times. Honey based products can be a good alternative as it is antibacterial and patient-friendly. We evaluated the bactericidal and cytotoxic effects of a honey based gel and silver sulphadiazine in a human burn wound model with Pseudomonas aeruginosa. After adding 10(5)colony forming units of P. aeruginosa, topical treatments were applied on the burn wound models. After 2, 12, 24, 28 and 70 h, bacteria were dislodged and counted by plating dilutions. Cytotoxic effects were evaluated histologically in samples of burn wound models treated topically for 3 weeks, without bacteria. L-Mesitran Soft significantly reduced the bacterial load (5-log reduction) up to 24h but did not completely eliminate bacteria from the burn wounds. After Flammazine(®) treatment, only a few colony forming units were observed at all time points. In contrast, re-epithelialization was significantly reduced after application of Flammazine(®) compared to L-Mesitran Soft or control. This in vitro model of burn wound infection can be used to evaluate topical treatments. L-Mesitran Soft is a good alternative for treating burn wounds but the slightly lower bactericidal activity in the burn wound model warrants a higher frequency of application. Copyright © 2012 Elsevier Ltd and ISBI. All rights reserved.

Digital Holographic Microscopy, a Method for Detection of Microorganisms in Plume Samples from Enceladus and Other Icy Worlds

PubMed Central

Bedrossian, Manuel; Lindensmith, Chris

2017-01-01

Abstract Detection of extant microbial life on Earth and elsewhere in the Solar System requires the ability to identify and enumerate micrometer-scale, essentially featureless cells. On Earth, bacteria are usually enumerated by culture plating or epifluorescence microscopy. Culture plates require long incubation times and can only count culturable strains, and epifluorescence microscopy requires extensive staining and concentration of the sample and instrumentation that is not readily miniaturized for space. Digital holographic microscopy (DHM) represents an alternative technique with no moving parts and higher throughput than traditional microscopy, making it potentially useful in space for detection of extant microorganisms provided that sufficient numbers of cells can be collected. Because sample collection is expected to be the limiting factor for space missions, especially to outer planets, it is important to quantify the limits of detection of any proposed technique for extant life detection. Here we use both laboratory and field samples to measure the limits of detection of an off-axis digital holographic microscope (DHM). A statistical model is used to estimate any instrument's probability of detection at various bacterial concentrations based on the optical performance characteristics of the instrument, as well as estimate the confidence interval of detection. This statistical model agrees well with the limit of detection of 103 cells/mL that was found experimentally with laboratory samples. In environmental samples, active cells were immediately evident at concentrations of 104 cells/mL. Published estimates of cell densities for Enceladus plumes yield up to 104 cells/mL, which are well within the off-axis DHM's limits of detection to confidence intervals greater than or equal to 95%, assuming sufficient sample volumes can be collected. The quantitative phase imaging provided by DHM allowed minerals to be distinguished from cells. Off-axis DHM's ability for rapid low-level bacterial detection and counting shows its viability as a technique for detection of extant microbial life provided that the cells can be captured intact and delivered to the sample chamber in a sufficient volume of liquid for imaging. Key Words: In situ life detection—Extant microorganisms—Holographic microscopy—Ocean Worlds—Enceladus—Imaging. Astrobiology 17, 913–925. PMID:28708412

Streptococcal adhesin SspA/B analogue peptide inhibits adherence and impacts biofilm formation of Streptococcus mutans

PubMed Central

Ito, Tatsuro; Ichinosawa, Takahiro; Shimizu, Takehiko

2017-01-01

Streptococcus mutans, the major causative agent of dental caries, adheres to tooth surfaces via the host salivary glycoprotein-340 (gp340). This adherence can be competitively inhibited by peptides derived from the SspA/B adhesins of Streptococcus gordonii, a human commensal microbe that competes for the same binding sites. Ssp(A4K-A11K), a double-lysine substituted SspA/B peptide analogue, has been shown to exhibit superior in vitro binding affinity for a gp340-derived peptide (SRCRP2), suggesting that Ssp(A4K-A11K) may be of clinical interest. In the present work, we tested the inhibitory effects of Ssp(A4K-A11K) on adherence and biofilm formation of S. mutans by reconstructing an artificial oral environment using saliva-coated polystyrene plates and hydroxyapatite disks. Bacterial adherence (adherence period: 1 h) was assessed by an enzyme-linked immunosorbent assay using biotinylated bacterial cells. Biofilm formation (periods: 8, 11, or 14 h) was assessed by staining and imaging of the sessile cells, or by recovering biofilm cells and plating for cell counts. The pH values of the culture media were measured as a biofilm acidogenicity indicator. Bactericidality was measured by loss of optical density during culturing in the presence of the peptide. We observed that 650 μM Ssp(A4K-A11K) significantly inhibited adherence of S. mutans to saliva-coated polystyrene; a similar effect was seen on bacterial affinity for SRCRP2. Ssp(A4K-A11K) had lesser effects on the adherence of commensal streptococci. Pretreatment of polystyrene and hydroxyapatite with 650 μM Ssp(A4K-A11K) significantly attenuated biofilm formation, whether tested with glucose- or sucrose-containing media. The SspA/B peptide’s activity did not reflect bactericidality. Strikingly, pH in Ssp-treated 8-h (6.8 ± 0.06) and 11-h (5.5 ± 0.06) biofilms showed higher values than the critical pH. Thus, Ssp(A4K-A11K) acts by inhibiting bacterial adherence and cariogrnic biofilm formation. We further consider these results in the context of the safety, specificity, and stability properties of the Ssp(A4K-A11K) peptide. PMID:28394940

Monitoring of bioaerosol inhalation risks in different environments using a six-stage Andersen sampler and the PCR-DGGE method.

PubMed

Xu, Zhenqiang; Yao, Maosheng

2013-05-01

Increasing evidences show that inhalation of indoor bioaerosols has caused numerous adverse health effects and diseases. However, the bioaerosol size distribution, composition, and concentration level, representing different inhalation risks, could vary with different living environments. The six-stage Andersen sampler is designed to simulate the sampling of different human lung regions. Here, the sampler was used in investigating the bioaerosol exposure in six different environments (student dorm, hospital, laboratory, hotel room, dining hall, and outdoor environment) in Beijing. During the sampling, the Andersen sampler was operated for 30 min for each sample, and three independent experiments were performed for each of the environments. The air samples collected onto each of the six stages of the sampler were incubated on agar plates directly at 26 °C, and the colony forming units (CFU) were manually counted and statistically corrected. In addition, the developed CFUs were washed off the agar plates and subjected to polymerase chain reaction (PCR)-denaturing gradient gel electrophoresis (DGGE) for diversity analysis. Results revealed that for most environments investigated, the culturable bacterial aerosol concentrations were higher than those of culturable fungal aerosols. The culturable bacterial and fungal aerosol fractions, concentration, size distribution, and diversity were shown to vary significantly with the sampling environments. PCR-DGGE analysis indicated that different environments had different culturable bacterial aerosol compositions as revealed by distinct gel band patterns. For most environments tested, larger (>3 μm) culturable bacterial aerosols with a skewed size distribution were shown to prevail, accounting for more than 60 %, while for culturable fungal aerosols with a normal size distribution, those 2.1-4.7 μm dominated, accounting for 20-40 %. Alternaria, Cladosporium, Chaetomium, and Aspergillus were found abundant in most environments studied here. Viable microbial load per unit of particulate matter was also shown to vary significantly with the sampling environments. The results from this study suggested that different environments even with similar levels of total microbial culturable aerosol concentrations could present different inhalation risks due to different bioaerosol particle size distribution and composition. This work fills literature gaps regarding bioaerosol size and composition-based exposure risks in different human dwellings in contrast to a vast body of total bioaerosol levels.

Development and test of photon-counting microchannel plate detector arrays for use on space telescopes

NASA Technical Reports Server (NTRS)

Timothy, J. G.

1976-01-01

The full sensitivity, dynamic range, and photometric stability of microchannel array plates(MCP) are incorporated into a photon-counting detection system for space operations. Components of the system include feedback-free MCP's for high gain and saturated output pulse-height distribution with a stable response; multi-anode readout arrays mounted in proximity focus with the output face of the MCP; and multi-layer ceramic headers to provide electrical interface between the anode array in a sealed detector tube and the associated electronics.

Interactions of Seedborne Bacterial Pathogens with Host and Non-Host Plants in Relation to Seed Infestation and Seedling Transmission

PubMed Central

Dutta, Bhabesh; Gitaitis, Ronald; Smith, Samuel; Langston, David

2014-01-01

The ability of seed-borne bacterial pathogens (Acidovorax citrulli, Clavibacter michiganensis subsp. michiganensis, Pseudomonas syringae pv. tomato, Xanthomonas euvesicatoria, and Pseudomonas syringae pv. glycinea) to infest seeds of host and non-host plants (watermelon, tomato, pepper, and soybean) and subsequent pathogen transmission to seedlings was investigated. A non-pathogenic, pigmented strain of Serratia marcescens was also included to assess a null-interacting situation with the same plant species. Flowers of host and non-host plants were inoculated with 1×106 colony forming units (CFUs)/flower for each bacterial species and allowed to develop into fruits or umbels (in case of onion). Seeds harvested from each host/non-host bacterial species combination were assayed for respective bacteria by plating on semi-selective media. Additionally, seedlots for each host/non-host bacterial species combination were also assayed for pathogen transmission by seedling grow-out (SGO) assays under greenhouse conditions. The mean percentage of seedlots infested with compatible and incompatible pathogens was 31.7 and 30.9% (by plating), respectively and they were not significantly different (P = 0.67). The percentage of seedlots infested with null-interacting bacterial species was 16.8% (by plating) and it was significantly lower than the infested lots generated with compatible and incompatible bacterial pathogens (P = 0.03). None of the seedlots with incompatible/null-interacting bacteria developed symptoms on seedlings; however, when seedlings were assayed for epiphytic bacterial presence, 19.5 and 9.4% of the lots were positive, respectively. These results indicate that the seeds of non-host plants can become infested with incompatible and null-interacting bacterial species through flower colonization and they can be transmitted via epiphytic colonization of seedlings. In addition, it was also observed that flowers and seeds of non-host plants can be colonized by compatible/incompatible/null-interacting bacteria to higher populations; however, the level of colonization differed significantly depending on the type of bacterial species used. PMID:24936863

Changes in bacterial diversity and community structure following pesticides addition to soil estimated by cultivation technique.

PubMed

Cycoń, Mariusz; Piotrowska-Seget, Zofia

2009-07-01

An experiment was conducted under laboratory conditions to investigate the effect of increasing concentrations of fenitrothion (2, 10 and 200 mg a.i./kg soil), diuron (1.5, 7.5 and 150 mg a.i./kg soil) and thiram (3.5, 17.5 and 350 mg a.i./kg soil) on soil respiration, bacterial counts and changes in culturable fraction of soil bacteria. To ascertain these changes, the community structure, bacterial biodiversity and process of colony formation, based on the r/K strategy concept, EP- and CD-indices and the FOR model, respectively, were determined. The results showed that the measured parameters were generally unaffected by the lowest dosages of pesticides, corresponding to the recommended field rates. The highest dosages of fenitrothion and thiram suppressed the peak SIR by 15-70% and 20-80%, respectively, while diuron increased respiration rate by 17-25% during the 28-day experiment. Also, the total numbers of bacteria increased in pesticide-treated soils. However, the reverse effect on day 1 and, in addition, in case of the highest dosages of insecticide on days 14 and 28, was observed. Analysis of the community structure revealed that in all soil treatments bacterial communities were generally dominated by K-strategists. Moreover, differences in the distribution of individual bacteria classes and the gradual domination of bacteria populations belonging to r-strategists during the experiment, as compared to control, was observed. However, on day 1, at the highest pesticide dosages, fast growing bacteria constituted only 1-10% of the total colonies number during 48 h of plate incubation, whereas in remaining samples they reached from 20 to 40% of total cfu. This effect, in case of fenitrothion, lasted till the end of the experiment. At the highest dosages of fenitrothion, diuron and at all dosages of thiram the decrease of biodiversity, as indicated by EP- and CD-indices on day 1, was found. At the next sampling time, no significant retarding or stimulating effect was detected. However, in case of CD values the higher differences were observed. The significant impact of pesticides on the physiological state of soil bacteria was not found. They were generally in dormant state (lambda < 0.5), but immediately after pesticides application, the additional reduction of frequency of bacterial cell proliferation (max. decrease of lambda value to 0.15 for thiram on day 14) and prolonged retardation time of colony appearance (max. increase of t(r) value to 1.39 for fenitrothion on day 1) on agar plates were found.

Sterilization of Escherichia coli O157:H7 using micro corona ionizer.

PubMed

Chua, Beelee; Son, Ahjeong

2014-06-01

We demonstrated in vitro sterilization of Escherichia coli O157:H7 bacteria on agar by a pin-between-planes micro corona ionizer. The gap between the pin and the grid was ~1.1 mm, the length of the grid was ~2.1 mm and the height was ~1.0 mm. The effective pin radius and discharge length were both approximated to be 200 μm. Ozone generation rates of ~2.3 × 10(-3) mg/s, ~2.7 × 10(-3) mg/s and ~3.5 × 10(-3) mg/s at 1,500 V were calculated for relative humidity (RH) of 35 %, 25 % and 10 % respectively. Analytical ozone generation rate increases as RH decreases and it is consistent with experimental observations. Using target and control petri dishes with E. coli plated agar, the sterilization capability of the micro corona ionizer at 37 °C for 24 h was evaluated. A ~60 % reduction in bacterial colony was shown with plate counting and its kill radius could be tuned from ~ 20 mm to ~5 mm by reducing the duty cycle from 100 % to 50 % with 30 min pulse width. The results suggested that the micro corona ionizer might be suitable as a tunable ozone source in wound dressing for chronic wound management.

GreenLight Model 960.

PubMed

Fernandes, Richard; Carey, Conn; Hynes, James; Papkovsky, Dmitri

2013-01-01

The importance of food safety has resulted in a demand for a more rapid, high-throughput method for total viable count (TVC). The industry standard for TVC determination (ISO 4833:2003) is widely used but presents users with some drawbacks. The method is materials- and labor-intensive, requiring multiple agar plates per sample. More importantly, the method is slow, with 72 h typically required for a definitive result. Luxcel Biosciences has developed the GreenLight Model 960, a microtiter plate-based assay providing a rapid high-throughput method of aerobic bacterial load assessment through analysis of microbial oxygen consumption. Results are generated in 1-12 h, depending on microbial load. The mix and measure procedure allows rapid detection of microbial oxygen consumption and equates oxygen consumption to microbial load (CFU/g), providing a simple, sensitive means of assessing the microbial contamination levels in foods (1). As bacteria in the test sample grow and respire, they deplete O2, which is detected as an increase in the GreenLight probe signal above the baseline level (2). The time required to reach this increase in signal can be used to calculate the CFU/g of the original sample, based on a predetermined calibration. The higher the initial microbial load, the earlier this threshold is reached (1).

Extensive subclinical sinusitis leading to Moraxella osloensis meningitis.

PubMed

Fox-Lewis, A; Coltart, G; Rice, S; Sen, R; Gourtsoyannis, Y; Hyare, H; Gupta, R K

2016-01-01

We report a case of a 31 year old male with extensive subclinical sinusitis leading to erosion in the cribriform plate and subsequent meningitis caused by the organism Moraxella osloensis . The patient presented to the emergency department with rapid onset confusion, neck stiffness and headache. Inflammatory markers, renal and liver function, and a chest radiograph were all normal. CT Head showed extensive polyp disease in the paranasal sinuses with expansion of the left frontal sinus and CT Sinuses revealed an area of low attenuation in the cribriform plate consistent with bony erosion. MRI Head showed thick loculated sinus inflammation. Lumbar puncture yielded CSF with a high white cell count of predominantly mononuclear cells, no visible organisms and an elevated protein. CSF microscopy, culture and viral PCR were not diagnostic, and so the CSF was sent for 16S rDNA PCR screening, which identified the rDNA of Moraxella osloensis . Moraxella osloensis is a rare cause of bacterial meningitis, with only a few reported cases. This case illustrates that sinusitis, while a common condition, when severe can predispose to intracranial infection with atypical and low virulence organisms such as Moraxella species, which do not commonly cause invasive CNS disease. This case represents the first case of Moraxella osloensis meningitis reported from the United Kingdom.

Evaluation of microbiological accumulation capability of the commercial sponge Spongia officinalis var. adriatica (Schmidt) (Porifera, Demospongiae).

PubMed

Stabili, Loredana; Licciano, Margherita; Longo, Caterina; Corriero, Giuseppe; Mercurio, Maria

2008-05-01

This study was carried out to evaluate the microbiological accumulation capability of the demosponge Spongia officinalis var. adriatica. Six microbiological parameters were researched in two sampling periods in the water and in reared sponge samples coming from sites with different degrees of microbial contamination: an off-shore fish farm displaced off the Apulian coast (Southern Adriatic Sea) and a no-impacted area displaced into the Marine Protected Area of Porto Cesareo (Apulian coast-Ionian Sea). We detected the density of culturable heterotrophic bacteria by spread plate on marine agar, total culturable bacteria at 37 degrees C on Plate Count Agar and vibrios on thiosulphate-citrate-bile-sucrose-salt (TCBS) agar. Total and fecal coliforms as well as fecal streptococci concentrations were detected by the MPN method. Bacterial densities were always higher in the sponge homogenates compared with the corresponding seawater in the sampling points and in both sampling periods. As regard vibrios, total culturable bacteria at 37 degrees C and fecal streptococci concentrations, the highest values were observed in the sponge samples coming from the off-shore fish farm during the summer period. The ability of Spongia officinalis var. adriatica to accumulate the microbial pollution indicators suggests that this species can be employed as a bioindicator for monitoring water quality.

Assessment of Chicken Carcass Microbiome Responses During Processing in the Presence of Commercial Antimicrobials Using a Next Generation Sequencing Approach

PubMed Central

Ae Kim, Sun; Hong Park, Si; In Lee, Sang; Owens, Casey M.; Ricke, Steven C.

2017-01-01

The purpose of this study was to 1) identify microbial compositional changes on chicken carcasses during processing, 2) determine the antimicrobial efficacy of peracetic acid (PAA) and Amplon (blend of sulfuric acid and sodium sulfate) at a poultry processing pilot plant scale, and 3) compare microbial communities between chicken carcass rinsates and recovered bacteria from media. Birds were collected from each processing step and rinsates were applied to estimate aerobic plate count (APC) and Campylobacter as well as Salmonella prevalence. Microbiome sequencing was utilized to identify microbial population changes over processing and antimicrobial treatments. Only the PAA treatment exhibited significant reduction of APC at the post chilling step while both Amplon and PAA yielded detectable Campylobacter reductions at all steps. Based on microbiome sequencing, Firmicutes were the predominant bacterial group at the phyla level with over 50% frequency in all steps while the relative abundance of Proteobacteria decreased as processing progressed. Overall microbiota between rinsate and APC plate microbial populations revealed generally similar patterns at the phyla level but they were different at the genus level. Both antimicrobials appeared to be effective on reducing problematic bacteria and microbiome can be utilized to identify optimal indicator microorganisms for enhancing product quality. PMID:28230180

Short communication: bulk tank total bacterial count in dairy sheep: factors of variation and relationship with somatic cell count.

PubMed

Gonzalo, C; Carriedo, J A; Beneitez, E; Juárez, M T; De La Fuente, L F; San Primitivo, F

2006-02-01

A total of 9,353 records for bulk tank total bacterial count (TBC) were obtained over 1 yr from 315 dairy ewe flocks belonging to the Sheep Improvement Consortium (CPO) in Castilla-León (Spain). Analysis of variance showed significant effects of flock, breed, month within flock, dry therapy, milking type and installation, and logSCC on logTBC. Flock and month within flock were important variation factors as they accounted for 22.0 and 22.1% of the variance, respectively. Considerable repeatability values were obtained for both random factors. Hand milking and bucket-milking machines elicited highest logTBC (5.31), whereas parlor systems with looped milkline (5.01) elicited the lowest logTBC. The implementation of dry therapy practice (5.12) showed significantly lower logTBC than when not used (5.25). Variability in logTBC among breeds ranged from 5.24 (Awassi) to 5.07 (Churra). However, clinical outbreaks of contagious agalactia did not increase TBC significantly. A statistically significant relationship was found between logTBC and logSCC, the correlation coefficient between the variables being r = 0.23. Programs for improving milk hygiene should be implemented for both total bacterial count and somatic cell count variables at the same time.

Identification of Bacterial Species in Kuwaiti Waters Through DNA Sequencing

NASA Astrophysics Data System (ADS)

Chen, K.

2017-01-01

With an objective of identifying the bacterial diversity associated with ecosystem of various Kuwaiti Seas, bacteria were cultured and isolated from 3 water samples. Due to the difficulties for cultured and isolated fecal coliforms on the selective agar plates, bacterial isolates from marine agar plates were selected for molecular identification. 16S rRNA genes were successfully amplified from the genome of the selected isolates using Universal Eubacterial 16S rRNA primers. The resulted amplification products were subjected to automated DNA sequencing. Partial 16S rDNA sequences obtained were compared directly with sequences in the NCBI database using BLAST as well as with the sequences available with Ribosomal Database Project (RDP).

Efficacy of Various Chemical Disinfectants on Biofilms Formed in Spacecraft Potable Water System Component

NASA Technical Reports Server (NTRS)

Wong, Willy; Garcia, Veronica; Castro, Victoria; Ott, Mark; Duane

2009-01-01

As the provision of potable water is critical for successful habitation of the International Space Station (ISS), life support systems were installed in December 2008 to recycle both humidity from the atmosphere and urine to conserve available water in the vehicle. Pre-consumption testing from the dispensing needle at the Potable Water Dispenser (PWD) indicated that bacterial concentrations exceeded the current ISS specifications of 50 colony forming units (CFU) per ml. Subsequent investigations revealed that a corrugated stainless steel flex hose upstream of the dispensing needle in the PWD was filled with non-sterile water and left at room temperature for over one month before launch. To simulate biofilm formation that was suspected in the flight system, sterile flex hoses were seeded with a consortium of bacterial isolates previously recovered from other ISS water systems, which included Ralstonia pickettii, Burkholderia multivorans, Caulobacter vibrioides., and Cupriavidus pauculus. After 5 days of incubation, these hoses were challenged with various chemical disinfectants including hydrogen peroxide, colloidal silver, and buffered pH solutions to determine the ability of the disinfectants to decrease and maintain bacterial concentrations below ISS specifications. Disinfection efficacy over time was measured by collecting daily heterotrophic plate counts following exposure to the disinfectants. A single flush with either 6% hydrogen peroxide solution or a mixture of 3% hydrogen peroxide and 400 ppb colloidal silver effectively reduced the bacterial concentrations to less than 1 CFU/ml for a period of up to 2 months. Testing results indicated that hydrogen peroxide and mixtures of hydrogen peroxide and colloidal silver have tremendous potential as alternative disinfectants for ISS water systems.

Reducing oyster-associated bacteria levels using supercritical fluid CO2 as an agent of warm pasteurization.

PubMed

Meujo, Damaris A F; Kevin, Dion A; Peng, Jiangnan; Bowling, John J; Liu, Jianping; Hamann, Mark T

2010-03-31

An innovative approach to Post-Harvest Processing (PHP) of oysters is introduced focusing on the effects of supercritical carbon dioxide (scCO(2)) on bacterial contaminants trapped in the digestive system of oysters. Oysters were exposed to scCO(2) under two conditions: (1) 100 bar and 37 degrees C for 30 min and (2) 172 bar and 60 degrees C for 60 min. Using FDA standard guidelines for food analysis, variations in the Aerobic Plate Count (APC) were assessed. It was established that exposing oysters to CO(2) at 100 bar and 37 degrees C for 30 min and at 172 bar and 60 degrees C for 60 min induced 2-log and 3-log reductions in the APC respectively. The decrease in the microbial load as a result of treatment with scCO(2) was found to be significant (P=0.002). A release of adductor muscles from the shell was noted in oysters treated at 172 bar and 60 degrees C for 60 min; this was not the case for oysters treated at 100 bar and 37 degrees C for 30 min. A blind study allowing sensory analysis of treated vs. untreated oysters was also completed and no significant change in the physical appearance, smell, or texture was recorded. In this paper, we also report the effect of scCO(2) on several bacterial isolates, including a referenced ATCC strain of a non-pathogenic Vibrio (Vibrio fischeri) as well as several other bacterial isolates cultured from oyster' tissues and found to share biochemical features common to pathogenic Vibrio strains. A complete inactivation (minimum 7-log reduction) was achieved with these latter bacterial isolates. A 6-log reduction was observed with V. fischeri. Copyright 2009 Elsevier B.V. All rights reserved.

Cultivation and qPCR Detection of Pathogenic and Antibiotic-Resistant Bacterial Establishment in Naive Broiler Houses.

PubMed

Brooks, J P; McLaughlin, M R; Adeli, A; Miles, D M

2016-05-01

Conventional commercial broiler production involves the rearing of more than 20,000 broilers in a single confined space for approximately 6.5 wk. This environment is known for harboring pathogens and antibiotic-resistant bacteria, but studies have focused on previously established houses with mature litter microbial populations. In the current study, a set of three naive houses were followed from inception through 11 broiler flocks and monitored for ambient climatic conditions, bacterial pathogens, and antibiotic resistance. Within the first 3 wk of the first flock cycle, 100% of litter samples were positive for and , whereas was cultivation negative but PCR positive. Antibiotic resistance genes were ubiquitously distributed throughout the litter within the first flock, approaching 10 to 10 genomic units g. Preflock litter levels were approximately 10 CFU g for heterotrophic plate count bacteria, whereas midflock levels were >10 colony forming units (CFU) g; other indicators demonstrated similar increases. The influence of intrahouse sample location was minor. In all likelihood, given that preflock levels were negative for pathogens and antibiotic resistance genes and 4 to 5 Log lower than flock levels for indicators, incoming birds most likely provided the colonizing microbiome, although other sources were not ruled out. Most bacterial groups experienced a cyclical pattern of litter contamination seen in other studies, whereas microbial stabilization required approximately four flocks. This study represents a first-of-its-kind view into the time required for bacterial pathogens and antibiotic resistance to colonize and establish in naive broiler houses. Copyright © by the American Society of Agronomy, Crop Science Society of America, and Soil Science Society of America, Inc.

Effect of postharvest UV-C treatment on the bacterial diversity of Ataulfo mangoes by PCR-DGGE, survival of E. coli and antimicrobial activity

PubMed Central

Fernández-Suárez, Rocío; Ramírez-Villatoro, Guadalupe; Díaz-Ruiz, Gloria; Eslava, Carlos; Calderón, Montserrat; Navarro-Ocaña, Arturo; Trejo-Márquez, Andrea; Wacher, Carmen

2013-01-01

Since Mexico is the second largest exporter of mangoes, its safety assurance is essential. Research in microbial ecology and knowledge of complex interactions among microbes must be better understood to achieve maximal control of pathogens. Therefore, we investigated the effect of UV-C treatments on bacterial diversity of the Ataulfo mangoes surface using PCR-DGGE analysis of variable region V3 of 16S rRNA genes, and the survival of E. coli, by plate counting. The UV-C irradiation reduced the microbial load on the surface of mangoes immediately after treatment and the structure of bacterial communities was modified during storage. We identified the key members of the bacterial communities on the surface of fruits, predominating Enterobacter genus. Genera as Lactococcus and Pantoea were only detected on the surface of non-treated (control) mangoes. This could indicate that these genera were affected by the UV-C treatment. On the other hand, the treatment did not have a significant effect on survival of E. coli. However, genera that have been recognized as antagonists against foodborne pathogens were identified in the bands patterns. Also, phenolic compounds were determined by HPLC and antimicrobial activity was assayed according to the agar diffusion method. The main phenolic compounds were chlorogenic, gallic, and caffeic acids. Mango peel methanol extracts (UV-C treated and control mangoes) showed antimicrobial activity against strains previously isolated from mango, detecting significant differences (P < 0.05) among treated and control mangoes after 4 and 12 days of storage. Ps. fluorescens and Ps. stutszeri were the most sensitive. PMID:23761788

Normalization of test and evaluation of biothreat detection systems: overcoming microbial air content fluctuations by using a standardized reagent bacterial mixture.

PubMed

Berchebru, Laurent; Rameil, Pascal; Gaudin, Jean-Christophe; Gausson, Sabrina; Larigauderie, Guilhem; Pujol, Céline; Morel, Yannick; Ramisse, Vincent

2014-10-01

Test and evaluation of engineered biothreat agent detection systems ("biodetectors") are a challenging task for government agencies and industries involved in biosecurity and biodefense programs. In addition to user friendly features, biodetectors need to perform both highly sensitive and specific detection, and must not produce excessive false alerts. In fact, the atmosphere displays a number of variables such as airborne bacterial content that can interfere with the detection process, thus impeding comparative tests when carried out at different times or places. To overcome these bacterial air content fluctuations, a standardized reagent bacterial mixture (SRBM), consisting in a collection of selected cultivable environmental species that are prevalent in temperate climate bioaerosols, was designed to generate a stable, reproducible, and easy to use surrogate of bioaerosol sample. The rationale, design, and production process are reported. The results showed that 8.59; CI 95%: 8.46-8.72 log cfu distributed into vials underwent a 0.95; CI 95%: 0.65-1.26 log viability decay after dehydration and subsequent reconstitution, thus advantageously mimicking a natural bioaerosol sample which is typically composed of cultivable and uncultivable particles. Dehydrated SRBM was stable for more than 12months at 4°C and allowed the reconstitution of a dead/live cells aqueous suspension that is stable for 96h at +4°C, according to plate counts. Specific detection of a simulating biothreat agent (e.g. Bacillus atrophaeus) by immuno-magnetic or PCR assays did not display any significant loss of sensitivity, false negative or positive results in the presence of SRBM. This work provides guidance on testing and evaluating detection devices, and may contribute to the establishment of suitable standards and normalized procedures. Copyright © 2014 Elsevier B.V. All rights reserved.

Reducing Oyster-Associated Bacteria Levels Using Supercritical Fluid CO2 as an Agent of Warm Pasteurization

PubMed Central

Meujo, Damaris A.F.; Kevin, Dion; Peng, Jiangnan; Bowling, John J.; Liu, Jianping; Hamann, Mark T.

2010-01-01

An innovative approach to Post-Harvest Processing (PHP) of oysters is introduced focusing on the effects of supercritical carbon dioxide (scCO2) on bacterial contaminants trapped in the digestive system of oysters. Oysters were exposed to scCO2 under two conditions: (1) 100 bar and 37 °C for 30 minutes and (2) 172 bar and 60 °C for 60 minutes. Using FDA standard guidelines for food analysis, variations in the Aerobic Plate Count (APC) was assessed. It was established that exposing oysters to CO2 at 100 bar and 37 °C for 30 minutes and at 172 bar and 60°C for 60 minutes induced 2-log and 3-log reductions in the APC respectively. The decrease in the microbial load as a result of treatment with scCO2 was found to be significant (P=0.002). A release of adductor muscles from the shell was noted in oysters treated at 172 bar and 60 °C for 60 minutes; this was not the case for oysters treated at 100 bar and 37 °C for 30 minutes. A blind study allowing sensory analysis of treated vs. untreated oysters was also completed and no significant change in the physical appearance, smell, or texture was recorded. In this paper, we also report the effect of scCO2 on several bacterial isolates, including a referenced ATCC strain of a non pathogenic Vibrio (V. fisherii) as well as several other bacterial isolates cultured from oyster’ tissues and found to share biochemical features common to pathogenic Vibrio strains. A complete inactivation (minimum 7-log reduction) was achieved with these latter bacterial isolates. A 6-log reduction was observed with V. fisherii. PMID:20022650

Inhibition of Biofilm Formation by Esomeprazole in Pseudomonas aeruginosa and Staphylococcus aureus

PubMed Central

Singh, Vandana; Arora, Vaneet; Alam, M. Jahangir

2012-01-01

Staphylococcus aureus and Pseudomonas aeruginosa are common nosocomial pathogens responsible for biofilm-associated infections. Proton pump inhibitors (PPI), such as esomeprazole, may have novel antimicrobial properties. The objective of this study was to assess whether esomeprazole prevents sessile bacterial growth and biofilm formation and whether it may have synergistic killing effects with standard antibiotics. The antibiofilm activity of esomeprazole at 0.25 mM was tested against two strains each of S. aureus and P. aeruginosa. Bacterial biofilms were prepared using a commercially available 96-peg-plate Calgary biofilm device. Sessile bacterial CFU counts and biomass were assessed during 72 hours of esomeprazole exposure. The killing activities after an additional 24 hours of vancomycin (against S. aureus) and meropenem (against P. aeruginosa) treatment with or without preexposure to esomeprazole were also assessed by CFU and biomass analyses. P. aeruginosa and S. aureus strains exposed to esomeprazole displayed decreased sessile bacterial growth and biomass (P < 0.001, each parameter). After 72 h of exposure, there was a 1-log10 decrease in the CFU/ml of esomeprazole-exposed P. aeruginosa and S. aureus strains compared to controls (P < 0.001). After 72 h of exposure, measured absorbance was 100% greater in P. aeruginosa control strains than in esomeprazole-exposed strains (P < 0.001). Increased killing and decreased biomass were observed for esomeprazole-treated bacteria compared to untreated controls exposed to conventional antibiotics (P < 0.001, each parameter). Reduced biofilm growth after 24 h was visibly apparent by light micrographs for P. aeruginosa and S. aureus isolates exposed to esomeprazole compared to untreated controls. In conclusion, esomeprazole demonstrated an antibiofilm effect against biofilm-producing S. aureus and P. aeruginosa. PMID:22664967

Statistical modeling of dental unit water bacterial test kit performance.

PubMed

Cohen, Mark E; Harte, Jennifer A; Stone, Mark E; O'Connor, Karen H; Coen, Michael L; Cullum, Malford E

2007-01-01

While it is important to monitor dental water quality, it is unclear whether in-office test kits provide bacterial counts comparable to the gold standard method (R2A). Studies were conducted on specimens with known bacterial concentrations, and from dental units, to evaluate test kit accuracy across a range of bacterial types and loads. Colony forming units (CFU) were counted for samples from each source, using R2A and two types of test kits, and conformity to Poisson distribution expectations was evaluated. Poisson regression was used to test for effects of source and device, and to estimate rate ratios for kits relative to R2A. For all devices, distributions were Poisson for low CFU/mL when only beige-pigmented bacteria were considered. For higher counts, R2A remained Poisson, but kits exhibited over-dispersion. Both kits undercounted relative to R2A, but the degree of undercounting was reasonably stable. Kits did not grow pink-pigmented bacteria from dental-unit water identified as Methylobacterium rhodesianum. Only one of the test kits provided results with adequate reliability at higher bacterial concentrations. Undercount bias could be estimated for this device and used to adjust test kit results. Insensitivity to methylobacteria spp. is problematic.

Is it safe to re-access sodium bicarbonate bottles for use in minor surgery?

PubMed

Bjornson, Lindsay; Bucevska, Marija; Tilley, Peter; Verchere, Cynthia

2018-04-06

Sodium bicarbonate is added to lidocaine to reduce injection pain. In Canada, it is available in vials exceeding the injection volume 100-fold. These are single-use vials that should be disposed of after one access. Some surgeons re-use vials to reduce waste, potentially causing contamination. This study aims to review the safety of sodium bicarbonate and assess alternatives to current practice. Strains of Staphylococcus aureus, Staphylococcus epidermidis, Pseudomonas aeruginosa, and Burkholderia cepacia were used to assess bacterial growth in vials of sodium bicarbonate. Each pathogen was inoculated into a vial for 14 days at room temperature. At several time points, 1 mL of solution was removed and diluted. One hundred microliters were transferred to blood agar plates and incubated at 35 °C. Colony counts were calculated, averaged and plotted onto a logarithmic graph. Colony counts of all strains fell below observational threshold after 7 days in sodium bicarbonate. Although all strains were reduced, bacteria can survive in sodium bicarbonate for several days, during which transmission may occur. Sodium bicarbonate vials should be treated as single-dose, as indicated by the manufacturers. To reduce waste, hospital pharmacies can repackage sodium bicarbonate into smaller vials or pre-alkalize lidocaine with sodium bicarbonate. Copyright © 2018 Elsevier Inc. All rights reserved.

Sustained release vancomycin-coated titanium alloy using a novel electrostatic dry powder coating technique may be a potential strategy to reduce implant-related infection.

PubMed

Han, Jing; Yang, Yi; Lu, Junren; Wang, Chenzhong; Xie, Youtao; Zheng, Xuebin; Yao, Zhenjun; Zhang, Chi

2017-07-24

In order to tackle the implant-related infection, a novel way was developed in this study to coat vancomycin particles mixed with controlled release coating materials onto the surface of titanium alloy by using an electrostatic dry powder coating technique. To characterize this sustained release antibacterial coating, surface morphology, in vitro and in vivo drug release were sequentially evaluated. In vitro cytotoxicity was tested by Cell Counting Kit-8 (CCK-8) assay and cytological changes were observed by inverted microscope. The antibacterial properties against MRSA, including a bacterial growth inhibition assay and a colony-counting test by spread plate method were performed. Results indicated that the vancomycin-coated sample was biocompatible for Human osteoblast cell line MG-63 and displayed effective antibacterial ability against MRSA. The coating film was revealed uniform by scanning electron microscopy. Both the in vitro and in vivo drug release kinetics showed an initially high release rate, followed by an extended period of sustained drug release over 7 days. These results suggest that with good biocompatibility and antibacterial ability, the sustained release antibacterial coating of titanium alloy using our novel electrostatic dry powder coating process may provide a promising candidate for the treatment of orthopedic implant-related infection.

Effect of Iranian Ziziphus honey on growth of some foodborne pathogens.

PubMed

Ekhtelat, Maryam; Ravaji, Karim; Parvari, Masood

2016-01-01

Honey has previously been shown to have wound healing and antimicrobial properties, but this is dependent on the type of honey, geographical location, and flower from which the final product is derived. We tested the antimicrobial activity of a natural honey originating from the Ziziphus spina-christi tree, against selected strains of bacteria. Ziziphus honey among more than a 100 verities of honey is known to have the greatest value of energy and minerals in it. The aim of this study was to determine the antibacterial activity of Ziziphus honey in 10%, 20%, 30%, and 40% dilutions (v/v) against Listeria monocytogenes, Salmonella typhimurium, Escherichia coli, and Staphylococcus aureus. Viable count enumeration of the sample was investigated after 0, 24, 72, and 120 h postinoculation with any of the bacteria using pour-plate method. The findings indicate that Ziziphus honey was effective against these pathogenic bacteria. In a comparative trial, antibacterial activity of Ziziphus honey was higher after 120 h incubation for each four bacteria in most dilutions. The microbial count showed 3-7.5 log reduction comparing with control after 120 h. Therefore, it is recommended using Ziziphus honey as a natural preservative and antibacterial agent. Also, it could potentially be used as therapeutic agents against bacterial infection particularly to the tested microorganisms.

An investigation of the antibacterial ability and cytotoxicity of a novel cu-bearing 317L stainless steel

NASA Astrophysics Data System (ADS)

Sun, Da; Xu, Dake; Yang, Chunguang; Shahzad, M. Babar; Sun, Ziqing; Xia, Jin; Zhao, Jinlong; Gu, Tingyue; Yang, Ke; Wang, Guixue

2016-07-01

In order to solve the challenging problem of microbial infections caused by microorganisms on medical implants, it is imperative to develop novel antimicrobial biomaterials. This work demonstrated that 317L-Cu stainless steel (SS), created by adding copper through a solution and aging heat treatment process, exhibited good antibacterial properties against staphylococcus aureus, achieving 2 log reduction of planktonic cells after 5 days of incubation. In this study, the antibacterial test was performed using the plate count method, the fluorescence cell staining method and the quantitative polymerase chain reaction (qPCR) method. It is well known that a high concentration of copper ion can lead to cytotoxicity. This work explored the cytotoxicity of 317L-Cu SS through real-time cell analysis (RTCA). Experimental results demonstrated that the 317L-Cu SS possessed a satisfactory antibacterial ability against S. aureus, and the antibacterial rate based on the reduction of sessile cell count reached 98.3% after 24-hour treatment. The bacterial adhesion and the biofilm thickness were considerably reduced by the 317L-Cu SS. The results of RTCA suggested that 317L-Cu SS did not introduce cytotoxicity to mouse cells, indicating its suitability as a medical implant material.

Long-term impact of farm management and crops on soil microorganisms assessed by combined DGGE and PLFA analyses

PubMed Central

Stagnari, Fabio; Perpetuini, Giorgia; Tofalo, Rosanna; Campanelli, Gabriele; Leteo, Fabrizio; Della Vella, Umberto; Schirone, Maria; Suzzi, Giovanna; Pisante, Michele

2014-01-01

In the present study, long-term organic and conventional managements were compared at the experimental field of Monsampolo del Tronto (Marche region, Italy) with the aim of investigating soil chemical fertility and microbial community structure. A polyphasic approach, combining soil fertility indicators with microbiological analyses (plate counts, PCR-denaturing gradient gel electrophoresis [DGGE] and phospholipid fatty acid analysis [PLFA]) was applied. Organic matter, N as well as some important macro and micronutrients (K, P, Mg, Mn, Cu, and Zn) for crop growth, were more available under organic management. Bacterial counts were higher in organic management. A significant influence of management system and management x crop interaction was observed for total mesophilic bacteria, nitrogen fixing bacteria and actinobacteria. Interestingly, cultivable fungi were not detected in all analyzed samples. PLFA biomass was higher in the organic and Gram positive bacteria dominated the microbial community in both systems. Even if fungal biomass was higher in organic management, fungal PCR-DGGE fingerprinting revealed that the two systems were very similar in terms of fungal species suggesting that 10 years were not enough to establish a new dynamic equilibrium among ecosystem components. A better knowledge of soil biota and in particular of fungal community structure will be useful for the development of sustainable management strategies. PMID:25540640

Adherence of coagulase-negative staphylococci to plastic tissue culture plates: a quantitative model for the adherence of staphylococci to medical devices.

PubMed Central

Christensen, G D; Simpson, W A; Younger, J J; Baddour, L M; Barrett, F F; Melton, D M; Beachey, E H

1985-01-01

The adherence of coagulase-negative staphylococci to smooth surfaces was assayed by measuring the optical densities of stained bacterial films adherent to the floors of plastic tissue culture plates. The optical densities correlated with the weight of the adherent bacterial film (r = 0.906; P less than 0.01). The measurements also agreed with visual assessments of bacterial adherence to culture tubes, microtiter plates, and tissue culture plates. Selected clinical strains were passed through a mouse model for foreign body infections and a rat model for catheter-induced endocarditis. The adherence measurements of animal passed strains remained the same as those of the laboratory-maintained parent strain. Spectrophotometric classification of coagulase-negative staphylococci into nonadherent and adherent categories according to these measurements had a sensitivity, specificity, and accuracy of 90.6, 80.8, and 88.4%, respectively. We examined a previously described collection of 127 strains of coagulase-negative staphylococci isolated from an outbreak of intravascular catheter-associated sepsis; strains associated with sepsis were more adherent than blood culture contaminants and cutaneous strains (P less than 0.001). We also examined a collection of 84 strains isolated from pediatric patients with cerebrospinal fluid (CSF) shunts; once again, pathogenic strains were more adherent than were CSF contaminants (P less than 0.01). Finally, we measured the adherence of seven endocarditis strains. As opposed to strains associated with intravascular catheters and CSF shunts, endocarditis strains were less adherent than were saprophytic strains of coagulase-negative staphylococci. The optical densities of bacterial films adherent to plastic tissue culture plates serve as a quantitative model for the study of the adherence of coagulase-negative staphylococci to medical devices, a process which may be important in the pathogenesis of foreign body infections. Images PMID:3905855

Hyperspectral scattering profiles for prediction of the microbial spoilage of beef

NASA Astrophysics Data System (ADS)

Peng, Yankun; Zhang, Jing; Wu, Jianhu; Hang, Hui

2009-05-01

Spoilage in beef is the result of decomposition and the formation of metabolites caused by the growth and enzymatic activity of microorganisms. There is still no technology for the rapid, accurate and non-destructive detection of bacterially spoiled or contaminated beef. In this study, hyperspectral imaging technique was exploited to measure biochemical changes within the fresh beef. Fresh beef rump steaks were purchased from a commercial plant, and left to spoil in refrigerator at 8°C. Every 12 hours, hyperspectral scattering profiles over the spectral region between 400 nm and 1100 nm were collected directly from the sample surface in reflection pattern in order to develop an optimal model for prediction of the beef spoilage, in parallel the total viable count (TVC) per gram of beef were obtained by classical microbiological plating methods. The spectral scattering profiles at individual wavelengths were fitted accurately by a two-parameter Lorentzian distribution function. TVC prediction models were developed, using multi-linear regression, on relating individual Lorentzian parameters and their combinations at different wavelengths to log10(TVC) value. The best predictions were obtained with r2= 0.96 and SEP = 0.23 for log10(TVC). The research demonstrated that hyperspectral imaging technique is a valid tool for real-time and non-destructive detection of bacterial spoilage in beef.

Oregano essential oil as an antimicrobial additive to detergent for hand washing and food contact surface cleaning.

PubMed

Rhoades, J; Gialagkolidou, K; Gogou, M; Mavridou, O; Blatsiotis, N; Ritzoulis, C; Likotrafiti, E

2013-10-01

To investigate the potential use of oregano essential oil as an antimicrobial agent in liquid soap for hand washing and for food contact surface cleaning. Oregano essential oil (O.E.O.) was emulsified in liquid detergent solution. This was challenge tested against a commercial antimicrobial soap in hand washing trials using natural flora. Soap with O.E.O. was as effective as the commercial antimicrobial soap at reducing aerobic plate count on the hands and more effective than plain soap with no additives. Cloths wetted with soap with O.E.O. were used to clean three different surfaces contaminated with four bacterial pathogens. For three of the four pathogens, the addition of 0·5% v/v O.E.O. to the soap solution enhanced cleaning performance and also reduced bacterial survival on the cloth after cleaning. Oregano essential oil (0·5%) is effective as an antimicrobial additive to detergent solutions for hand washing and surface cleaning. This preliminary study has shown that oregano essential oil is a potential alternative to antimicrobials used in various detergents, such as chloroxylenol and triclosan, which can have adverse environmental and health effects. Further development could lead to a commercial product. © 2013 The Society for Applied Microbiology.

Black Currant (Ribes nigrum L.) and Bilberry (Vaccinium myrtillus L.) Fruit Juices Inhibit Adhesion of Asaia spp.

PubMed Central

2016-01-01

The aim of the study was to evaluate the activity of high-polyphenolic black currant (Ribes nigrum L.) and bilberry (Vaccinium myrtillus L.) juices against bacterial strains Asaia lannensis and Asaia bogorensis isolated as spoilage of commercial soft drinks. The composition of fruit juices was evaluated using chromatographic techniques HPLC and LC-MS. The adhesion to glass, polystyrene, and polyethylene terephthalate in two different culture media was evaluated by luminometry and the plate count method. The major anthocyanins in the V. myrtillus were petunidin-3-glucoside, malvidin-3-glucoside, cyanidin-3-glucoside, and delphinidin-3-glucoside, while in R. nigrum delphinidin-3-rutinoside and cyanidin-3-rutinoside were detected. The LC-MS analysis showed presence of anthocyanins (delphinidin, cyanidin, petunidin, and malvidin derivatives), phenolic acids (chlorogenic and neochlorogenic acids), flavonols (quercetin-3-glucoside, quercetin-3-rutinoside), and flavanols (procyanidin B2 and procyanidin type A2). Additionally, in the bilberry juice A type procyanidin trimer was detected. The adhesion of Asaia spp. cells depended on the type of medium, carbon sources, and the type of abiotic surfaces. We noted that the adhesion was significantly stronger in minimal medium containing sucrose. The addition of bilberry and black currant juices notably reduced bacterial growth as well as cell adhesion to polyethylene terephthalate surfaces. PMID:27747228

Rainfall simulation in greenhouse microcosms to assess bacterial-associated runoff from land-applied poultry litter.

PubMed

Brooks, John P; Adeli, Ardeshir; Read, John J; McLaughlin, Michael R

2009-01-01

Runoff water following a rain event is one possible source of environmental contamination after a manure application. This greenhouse study used a rainfall simulator to determine bacterial-associated runoff from troughs of common bermudagrass [Cynodon dactylon (L.) Pers.] that were treated with P-based, N-based, and N plus lime rates of poultry (Gallus gallus) litter, recommended inorganic fertilizer, and control. Total heterotrophic plate count (HPC) bacteria, total and thermotolerant coliforms, enterococci, staphylococci, Clostridium perfringens, Salmonella, and Campylobacter, as well as antibiotic resistance profiles for the staphylococci and enterococci isolates were all monitored in runoff waters. Analysis following five rainfall events indicated that staphylococci, enterococci, and clostridia levels were related to manure application rate. Runoff release of staphylococci, enterococci, and C. perfringens were approximately 3 to 6 log10 greater in litter vs. control treatment. In addition, traditional indicators such as thermotolerant and total coliforms performed poorly as fecal indicators. Some isolated enterococci demonstrated increased antibiotic resistance to polymixin b and/or select aminoglyocosides, while many staphylococci were susceptible to most antimicrobials tested. Results indicated poultry litter application can lead to microbial runoff following simulated rain events. Future studies should focus on the use of staphylococci, enterococci, and C. perfringens as indicators.

Growth on solid media.

PubMed

Elbing, Karen; Brent, Roger

2002-08-01

Detailed protocols are provided for titering and isolating bacterial colonies by serial dilutions, or alternatively by streaking or spreading a plate. Support protocols describe replica plating as well as methods for storing strains as agar stabs or frozen glycerol stocks.

Strain Library Imaging Protocol for high-throughput, automated single-cell microscopy of large bacterial collections arrayed on multiwell plates.

PubMed

Shi, Handuo; Colavin, Alexandre; Lee, Timothy K; Huang, Kerwyn Casey

2017-02-01

Single-cell microscopy is a powerful tool for studying gene functions using strain libraries, but it suffers from throughput limitations. Here we describe the Strain Library Imaging Protocol (SLIP), which is a high-throughput, automated microscopy workflow for large strain collections that requires minimal user involvement. SLIP involves transferring arrayed bacterial cultures from multiwell plates onto large agar pads using inexpensive replicator pins and automatically imaging the resulting single cells. The acquired images are subsequently reviewed and analyzed by custom MATLAB scripts that segment single-cell contours and extract quantitative metrics. SLIP yields rich data sets on cell morphology and gene expression that illustrate the function of certain genes and the connections among strains in a library. For a library arrayed on 96-well plates, image acquisition can be completed within 4 min per plate.

Selective enumeration of propionibacteria in Emmental-type cheese using Petrifilm™ aerobic count plates added to lithium glycerol broth.

PubMed

de Freitas, Rosângela; Luiz, Lívia M Pinheiro; Alves, Maura Pinheiro; Valence-Bertel, Florence; Nero, Luís Augusto; de Carvalho, Antônio Fernandes

2013-08-01

Propionibacteria derived from dairy products are relevant starter cultures for the production of Swiss and Emmental-type cheeses, and the monitoring of which is mandatory for proper quality control. This study aimed to evaluate an alternative procedure to enumerate propionibacteria, in order to develop a reliable and practical methodology to be employed by dairy industries. 2,3,5-triphenyltetrazolium chloride (TTC) inhibitory activity was tested against five reference strains (CIRM 09, 38, 39, 40 and 116); TTC at 0·0025% (w/v) was not inhibitory, with the exception of one strain (CIRM 116). Subsequently, the four TTC-resistant strains, three commercial starter cultures (PS-1, PB-I, and CHOO) and twelve Emmental-type cheese samples were subjected to propionibacteria enumeration using Lithium Glycerol (LG) agar, and Petrifilm™ Aerobic Count (AC) plates added to LG broth (anaerobic incubation at 30 °C for 7 d). Petrifilm™ AC added to LG broth presented high counts than LG agar (P<0·05) for only two reference strains (CIRM 39, and 40) and for all commercial starter cultures. Cheese sample counts obtained by both procedures did not show significant differences (P<0·05). Significant correlation indexes were observed between the counts recorded by both methods (P<0·05). These results demonstrate the reliability of Petrifilm™ AC plates added to LG broth in enumerating select Propionibacterium spp., despite some limitations observed for specific commercial starter cultures.

Optimization of high count rate event counting detector with Microchannel Plates and quad Timepix readout

NASA Astrophysics Data System (ADS)

Tremsin, A. S.; Vallerga, J. V.; McPhate, J. B.; Siegmund, O. H. W.

2015-07-01

Many high resolution event counting devices process one event at a time and cannot register simultaneous events. In this article a frame-based readout event counting detector consisting of a pair of Microchannel Plates and a quad Timepix readout is described. More than 104 simultaneous events can be detected with a spatial resolution of 55 μm, while >103 simultaneous events can be detected with <10 μm spatial resolution when event centroiding is implemented. The fast readout electronics is capable of processing >1200 frames/sec, while the global count rate of the detector can exceed 5×108 particles/s when no timing information on every particle is required. For the first generation Timepix readout, the timing resolution is limited by the Timepix clock to 10-20 ns. Optimization of the MCP gain, rear field voltage and Timepix threshold levels are crucial for the device performance and that is the main subject of this article. These devices can be very attractive for applications where the photon/electron/ion/neutron counting with high spatial and temporal resolution is required, such as energy resolved neutron imaging, Time of Flight experiments in lidar applications, experiments on photoelectron spectroscopy and many others.

Direct quantification of bacterial biomass in influent, effluent and activated sludge of wastewater treatment plants by using flow cytometry.

PubMed

Foladori, P; Bruni, L; Tamburini, S; Ziglio, G

2010-07-01

A rapid multi-step procedure, potentially amenable to automation, was proposed for quantifying viable and active bacterial cells, estimating their biovolume using flow cytometry (FCM) and to calculate their biomass within the main stages of a wastewater treatment plant: raw wastewater, settled wastewater, activated sludge and effluent. Fluorescent staining of bacteria using SYBR-Green I + Propidium Iodide (to discriminate cell integrity or permeabilisation) and BCECF-AM (to identify enzymatic activity) was applied to count bacterial cells by FCM. A recently developed specific procedure was applied to convert Forward Angle Light Scatter measured by FCM into the corresponding bacterial biovolume. This conversion permits the calculation of the viable and active bacterial biomass in wastewater, activated sludge and effluent, expressed as Volatile Suspended Solids (VSS) or particulate Chemical Oxygen Demand (COD). Viable bacterial biomass represented only a small part of particulate COD in raw wastewater (4.8 +/- 2.4%), settled wastewater (10.7 +/- 3.1%), activated sludge (11.1 +/- 2.1%) and effluent (3.2 +/- 2.2%). Active bacterial biomass counted for a percentage of 30-47% of the viable bacterial biomass within the stages of the wastewater treatment plant. Copyright 2010 Elsevier Ltd. All rights reserved.

Soybean extracts facilitate bacterial agglutination and prevent biofilm formation on orthodontic wire.

PubMed

Lee, Heon-Jin; Kwon, Tae-Yub; Kim, Kyo-Han; Hong, Su-Hyung

2014-01-01

Soybean is an essential food ingredient that contains a class of organic compounds known as isoflavones. It is also well known that several plant agglutinins interfere with bacterial adherence to smooth surfaces. However, little is known about the effects of soybean extracts or genistein (a purified isoflavone from soybean) on bacterial biofilm formation. We evaluated the effects of soybean (Glycine max) extracts, including fermented soybean and genistein, on streptococcal agglutination and attachment onto stainless steel orthodontic wire. After cultivating streptococci in biofilm medium containing soybean extracts and orthodontic wire, the viable bacteria attached to the wire were counted. Phase-contrast microscopy and scanning electron microscopy (SEM) analyses were conducted to evaluate bacterial agglutination and attachment. Our study showed that soybean extracts induce agglutination between streptococci, which results in bacterial precipitation. Conversely, viable bacterial counting and SEM image analysis of Streptococcus mutans attached to the orthodontic wire show that bacterial attachment decreases significantly when soybean extracts were added. However, there was no significant change in pre-attached S. mutans biofilm in response to soybean. A possible explanation for these results is that increased agglutination of planktonic streptococci by soybean extracts results in inhibition of bacterial attachment onto the orthodontic wire.

Bacterial adenosine triphosphate as a measure of urinary tract infection

NASA Technical Reports Server (NTRS)

Chappelle, E. W.; Picciolo, G. L.

1971-01-01

Procedure detects and counts bacteria present in urine samples. Method also determines bacterial levels in other aqueous body fluids including lymph fluid, plasma, blood, spinal fluid, saliva and mucous.

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.

Antimicrobial peptides secreted by equine mesenchymal stromal cells inhibit the growth of bacteria commonly found in skin wounds.

PubMed

Harman, Rebecca M; Yang, Steven; He, Megan K; Van de Walle, Gerlinde R

2017-07-04

The prevalence of chronic skin wounds in humans is high, and treatment is often complicated by the presence of pathogenic bacteria. Therefore, safe and innovative treatments to reduce the bacterial load in cutaneous wounds are needed. Mesenchymal stromal cells (MSC) are known to provide paracrine signals that act on resident skin cells to promote wound healing, but their potential antibacterial activities are not well described. The present study was designed to examine the antibacterial properties of MSC from horses, as this animal model offers a readily translatable model for MSC therapies in humans. Specifically, we aimed to (i) evaluate the in vitro effects of equine MSC on the growth of representative gram-negative and gram-positive bacterial species commonly found in skin wounds and (ii) define the mechanisms by which MSC inhibit bacterial growth. MSC were isolated from the peripheral blood of healthy horses. Gram-negative E. coli and gram-positive S. aureus were cultured in the presence of MSC and MSC conditioned medium (CM), containing all factors secreted by MSC. Bacterial growth was measured by plating bacteria and counting viable colonies or by reading the absorbance of bacterial cultures. Bacterial membrane damage was detected by incorporation of N-phenyl-1-naphthylamine (NPN). Antimicrobial peptide (AMP) gene and protein expression by equine MSC were determined by RT-PCR and Western blot analysis, respectively. Blocking of AMP activity of MSC CM was achieved using AMP-specific antibodies. We found that equine MSC and MSC CM inhibit the growth of E. coli and S. aureus, and that MSC CM depolarizes the cell membranes of these bacteria. In addition, we found that equine MSC CM contains AMPs, and blocking these AMPs with antibodies reduces the effects of MSC CM on bacteria. Our results demonstrate that equine MSC inhibit bacterial growth and secrete factors that compromise the membrane integrity of bacteria commonly found in skin wounds. We also identified four specific AMPs produced by equine MSC. The secretion of AMPs may contribute to the value of MSC as a therapy for cutaneous wounds in both horses and humans.

Hybrid Ion-Detector/Data-Acquisition System for a TOF-MS

NASA Technical Reports Server (NTRS)

Burton, William D., Jr.; Schultz, J. Albert; Vaughn, Valentine; McCully, Michael; Ulrich, Steven; Egan, Thomas F.

2006-01-01

A modified ion-detector/data-acquisition system has been devised to increase the dynamic range of a time-of-flight mass spectrometer (TOF-MS) that, previously, included a microchannel-plate detector and a data-acquisition system based on counting pulses and time-tagging them by use of a time-to-digital converter (TDC). The dynamic range of the TOF-MS was limited by saturation of the microchannel plate detector, which can handle no more than a few million counts per second. The modified system includes (1) a combined microchannel plate/discrete ion multiplier and (2) a hybrid data-acquisition system that simultaneously performs analog current or voltage measurements and multianode single-ion-pulse-counting time-of-flight measurements to extend the dynamic range of a TDC into the regime in which a mass peak comprises multiple ions arriving simultaneously at the detector. The multianode data are used to determine, in real time, whether the detector is saturated. When saturation is detected, the data-acquisition system selectively enables circuitry that simultaneously determines the ion-peak intensity by measuring the time profile of the analog current or voltage detector-output signal.

Detection of microbial concentration in ice-cream using the impedance technique.

PubMed

Grossi, M; Lanzoni, M; Pompei, A; Lazzarini, R; Matteuzzi, D; Riccò, B

2008-06-15

The detection of microbial concentration, essential for safe and high quality food products, is traditionally made with the plate count technique, that is reliable, but also slow and not easily realized in the automatic form, as required for direct use in industrial machines. To this purpose, the method based on impedance measurements represents an attractive alternative since it can produce results in about 10h, instead of the 24-48h needed by standard plate counts and can be easily realized in automatic form. In this paper such a method has been experimentally studied in the case of ice-cream products. In particular, all main ice-cream compositions of real interest have been considered and no nutrient media has been used to dilute the samples. A measurement set-up has been realized using benchtop instruments for impedance measurements on samples whose bacteria concentration was independently measured by means of standard plate counts. The obtained results clearly indicate that impedance measurement represents a feasible and reliable technique to detect total microbial concentration in ice-cream, suitable to be implemented as an embedded system for industrial machines.

Vancomycin-modified implant surface inhibits biofilm formation and supports bone-healing in an infected osteotomy model in sheep: a proof-of-concept study.

PubMed

Stewart, Suzanne; Barr, Stephanie; Engiles, Julie; Hickok, Noreen J; Shapiro, Irving M; Richardson, Dean W; Parvizi, Javad; Schaer, Thomas P

2012-08-01

Implant-associated infections contribute to patient morbidity and health care costs. We hypothesized that surface modification of titanium fracture hardware with vancomycin would support bone-healing and prevent bacterial colonization of the implant in a large-animal model. A unilateral transverse mid-diaphyseal tibial osteotomy was performed and repaired with a titanium locking compression plate in nine sheep. Four control animals were treated with an unmodified plate and five experimental animals were treated with a vancomycin-modified plate. The osteotomy was inoculated with 2.5 × 106 colony-forming units of Staphylococcus aureus. The animals were killed at three months postoperatively, and implants were retrieved aseptically. Microbiologic and histologic analyses, scanning electron and confocal microscopy, and microcomputed tomography were performed. All animals completed the study. Compared with the treatment cohort, control animals exhibited protracted lameness in the operatively treated leg. Gross findings during necropsy were consistent with an infected osteotomy accompanied by a florid and lytic callus. Microcomputed tomography and histologic analysis of the tibiae further supported the presence of septic osteomyelitis in the control cohort. Thick biofilms were also evident, and bacterial cultures were positive for Staphylococcus aureus in three of four control animals. In contrast, animals treated with vancomycin-treated plates exhibited a healed osteotomy site with homogenous remodeling, there was no evidence of biofilm formation on the retrieved plate, and bacterial cultures from only one of five animals were positive for Staphylococcus aureus. Vancomycin-derivatized plate surfaces inhibited implant colonization with Staphylococcus aureus and supported bone-healing in an infected large-animal model.

A non-invasive in vivo imaging system to study dissemination of bioluminescent Yersinia pestis CO92 in a mouse model of pneumonic plague.

PubMed

Sha, Jian; Rosenzweig, Jason A; Kirtley, Michelle L; van Lier, Christina J; Fitts, Eric C; Kozlova, Elena V; Erova, Tatiana E; Tiner, Bethany L; Chopra, Ashok K

2013-02-01

The gold standard in microbiology for monitoring bacterial dissemination in infected animals has always been viable plate counts. This method, despite being quantitative, requires sacrificing the infected animals. Recently, however, an alternative method of in vivo imaging of bioluminescent bacteria (IVIBB) for monitoring microbial dissemination within the host has been employed. Yersinia pestis is a Gram-negative bacterium capable of causing bubonic, septicemic, and pneumonic plague. In this study, we compared the conventional counting of bacterial colony forming units (cfu) in the various infected tissues to IVIBB in monitoring Y. pestis dissemination in a mouse model of pneumonic plague. By using a transposon mutagenesis system harboring the luciferase (luc) gene, we screened approximately 4000 clones and obtained a fully virulent, luc-positive Y. pestis CO92 (Y. pestis-luc2) reporter strain in which transposition occurred within the largest pMT1 plasmid which possesses murine toxin and capsular antigen encoding genes. The aforementioned reporter strain and the wild-type CO92 exhibited similar growth curves, formed capsule based on immunofluorescence microscopy and flow cytometry, and had a similar LD(50). Intranasal infection of mice with 15 LD(50) of CO92-luc2 resulted in animal mortality by 72 h, and an increasing number of bioluminescent bacteria were observed in various mouse organs over a 24-72 h period when whole animals were imaged. However, following levofloxacin treatment (10 mg/kg/day) for 6 days 24 h post infection, no luminescence was observed after 72 h of infection, indicating that the tested antimicrobial killed bacteria preventing their detection in host peripheral tissues. Overall, we demonstrated that IVIBB is an effective and non-invasive way of monitoring bacterial dissemination in animals following pneumonic plague having strong correlation with cfu, and our reporter CO92-luc2 strain can be employed as a useful tool to monitor the efficacy of antimicrobial countermeasures in real time. Copyright © 2012 Elsevier Ltd. All rights reserved.

Leuconostoc gasicomitatum is the dominating lactic acid bacterium in retail modified-atmosphere-packaged marinated broiler meat strips on sell-by-day.

PubMed

Susiluoto, Tuija; Korkeala, Hannu; Björkroth, K Johanna

2003-01-15

Lactic acid bacteria (LAB) in retail, modified-atmosphere-packaged (MAP), marinated broiler meat strips on sell-by-day were mainly identified as Leuconostoc gasicomitatum. A total of 32 packages, three to five packages of seven differently marinated broiler meat products, were studied at the end of the producer-defined shelf life (at 6 degrees C, 7-9 days depending on the manufacturer). Prior to the microbiological analyses, appearance and smell of the product was checked and pH measured. Bacteria were cultured on MRS and Tomato Juice Agar (TJA), Rogosa SL agar (SLA), Plate Count Agar (PCA) and Streptomycin Thallium Acetate Agar (STAA) for the enumeration of LAB, lactobacilli, total bacterial count and Brochothrix thermosphacta, respectively. The average CFU/g of the 32 packages was 2.3 x 10(8) on PCA. The highest bacterial average, 3.1 x 10(8), was recovered on TJA, the corresponding CFU/g averages on MRS and SLA being 2.3 x 10(8) and 1.3 x 10(8), respectively. Despite the high LAB numbers detected, radical spoilage changes such as unpleasant odor, slime production and formation of gas were not seen. B. thermosphacta did not form a significant part of the bacterial population since none of the levels exceeded the spoilage threshold level of 10(5) CFU/g reported in previous studies for this organism. In order to characterize the dominating LAB population, as many as 85, 85 and 88 colonies from MRS, TJA and SLA, respectively, were randomly picked and cultured pure. LAB were identified to species level using a 16 and 23S rDNA HindIiI RFLP (ribotyping) database. Fifty-six of the 170 isolates picked from the non-selective LAB media (MRS and TJA) were identified as L. gasicomitatum, followed by Carnobacterium divergens (41 isolates), Lactobacillus sakei and Lactobacillus curvatus subsp. melibiosus (31 isolates) and L. curvatus subsp. curvatus (20 isolates) species. SLA proved not to be completely selective for lactobacilli because the growth of Leuconostoc spp. was not inhibited, Carnobacterium spp. were the only species not detected on SLA.

Transverse Chemotactic Migration of Bacteria from High to Low Permeability Regions in a Dual Permeability Porous Microfluidic Device

NASA Astrophysics Data System (ADS)

Singh, R.; Olson, M. S.

2011-12-01

Low permeability regions sandwiched between high permeability regions such as clay lenses are difficult to treat using conventional treatment methods. Trace concentrations of contaminants such as non-aqueous phase liquids (NAPLs) remain trapped in these regions and over the time diffuse out into surrounding water thereby acting as a long term source of groundwater contamination. Bacterial chemotaxis (directed migration toward a contaminant source), may be helpful in enhancing bioremediation of such contaminated sites. This study is focused on simulating a two-dimensional dual-permeability groundwater contamination scenario using microfluidic devices and evaluating transverse chemotactic migration of bacteria from high to low permeability regions. A novel bi-layer polydimethylsiloxane (PDMS) microfluidic device was fabricated using photolithography and soft lithography techniques to simulate contamination of a dual- permeability region due to leakage from an underground storage tank into a low permeability region. This device consists of a porous channel through which a bacterial suspension (Escherchia Coli HCB33) is flown and another channel for injecting contaminant/chemo-attractant (DL-aspertic acid) into the porous channel. The pore arrangement in the porous channel contains a 2-D low permeability region surrounded by high permeability regions on both sides. Experiments were performed under chemotactic and non-chemotactic (replacing attractant with buffer solution in the non porous channel) conditions. Images were captured in transverse pore throats at cross-sections 4.9, 9.8, and 19.6 mm downstream from the attractant injection point and bacteria were enumerated in the middle of each pore throat. Bacterial chemotaxis was quantified in terms of the change in relative bacterial counts in each pore throat at cross-sections 9.8 and 19.6 mm with respect to counts at the cross-section at 4.9 mm. Under non-chemotactic conditions, relative bacterial count was observed to decrease at 9.8 mm and 19.6 mm cross-sections in low permeability regions due to dilution with the injectate from the non-porous channel (Figure 1). However, relative bacterial counts increased in the low permeability region at both downstream cross-sections under chemotactic conditions. A large increase in relative bacterial count in the pore throats just outside the low permeability region was also observed at both cross-sections (Figure 1). The bacterial chemotactic response was observed to decrease linearly with increasing Darcy velocity and at flow rate 0.220 mm/s the chemotactic effect was offset by the advective flow in the porous channel.

Influence of short distance transportation on tracheal bacterial content and lower airway cytology in horses.

PubMed

Allano, Marion; Labrecque, Olivia; Rodriguez Batista, Edisleidy; Beauchamp, Guy; Bédard, Christian; Lavoie, Jean-Pierre; Leclere, Mathilde

2016-08-01

The aim of this study was to determine the effects of short distance transportation on airway mucus, cytology and bacterial culture to identify potential biases in the diagnosis of airway diseases in referral centres. Eight healthy adult horses were studied using a prospective cross-over design. Mucus scores, tracheal wash (cytology, bacterial culture) and bronchoalveolar lavage fluid (BALF; cytology) were obtained while stabled and following 2.5 h transportation (with and without hay). Neutrophil counts, percentages and BALF neutrophilia frequency increased following transport without hay (P <0.05). No effect was observed on tracheal cytology and bacterial count (P > 0.05). BALF neutrophilia could develop solely as a result of transportation or due to interactions between repeated transports, ambient temperature, head position or other environmental factors. Copyright © 2016 Elsevier Ltd. All rights reserved.

In vitro assessments on bacterial adhesion and corrosion performance of TiN coating on Ti6Al4V titanium alloy synthesized by multi-arc ion plating

NASA Astrophysics Data System (ADS)

Lin, Naiming; Huang, Xiaobo; Zhang, Xiangyu; Fan, Ailan; Qin, Lin; Tang, Bin

2012-07-01

TiN coating was synthesized on Ti6Al4V titanium alloy surface by multi-arc ion plating (MIP) technique. Surface morphology, cross sectional microstructure, elemental distributions and phase compositions of the obtained coating were analyzed by means of scanning electron microscope (SEM), optical microscope (OM), glow discharge optical emission spectroscope (GDOES) and X-ray diffraction (XRD). Bacterial adhesion and corrosion performance of Ti6Al4V and the TiN coating were assessed via in vitro bacterial adhesion tests and corrosion experiments, respectively. The results indicated that continuous and compact coating which was built up by pure TiN with a typical columnar crystal structure has reached a thickness of 1.5 μm. This TiN coating could significantly reduce the bacterial adhesion and enhance the corrosion resistance of Ti6Al4V substrate.

A case-control study of domestic kitchen microbiology and sporadic Salmonella infection.

PubMed

Parry, S M; Slader, J; Humphrey, T; Holmes, B; Guildea, Z; Palmer, S R

2005-10-01

The microbiology of domestic kitchens in the homes of subjects who had suffered sporadic Salmonella infection (cases) was compared with control domestic kitchens. Case and control dishcloths and refrigerator swabs were examined for the presence of Salmonella spp., total Enterobacteriaceae counts and total aerobic colony counts. Salmonella spp. were isolated from both case and control dishcloths and refrigerators but there were no significant differences between the two groups. Colony counts were similar in case and control dishcloths and refrigerator swabs. There was no relationship between the total counts and presence of Salmonella . There was no evidence that cases of Salmonella infection were more likely to have kitchens which were contaminated with these bacteria or have higher bacterial counts than controls. Total bacterial counts were poor indicators of Salmonella contamination of the domestic kitchen environment. Further factors which could not be identified by a study of this design may increase risk of Salmonella food poisoning. These factors may include individual susceptibility of the patient. Alternatively, sporadic cases of Salmonella food poisoning may arise from food prepared outside the home.

A novel method of personnel cooling in an operating theatre environment.

PubMed

Casha, Aaron R; Manché, Alexander; Camilleri, Liberato; Gauci, Marilyn; Grima, Joseph N; Borg, Michael A

2014-10-01

An optimized theatre environment, including personal temperature regulation, can help maintain concentration, extend work times and may improve surgical outcomes. However, devices, such as cooling vests, are bulky and may impair the surgeon's mobility. We describe the use of a low-cost, low-energy 'bladeless fan' as a personal cooling device. The safety profile of this device was investigated by testing air quality using 0.5- and 5-µm particle counts as well as airborne bacterial counts on an operating table simulating a wound in a thoracic operation in a busy theatre environment. Particle and bacterial counts were obtained with both an empty and full theatre, with and without the 'bladeless fan'. The use of the 'bladeless fan' within the operating theatre during the simulated operation led to a minor, not statistically significant, lowering of both the particle and bacterial counts. In conclusion, the 'bladeless fan' is a safe, effective, low-cost and low-energy consumption solution for personnel cooling in a theatre environment that maintains the clean room conditions of the operating theatre. © The Author 2014. Published by Oxford University Press on behalf of the European Association for Cardio-Thoracic Surgery. All rights reserved.

Comparison of a chlorhexidine and a benzoyl peroxide shampoo as sole treatment in canine superficial pyoderma.

PubMed

Loeffler, A; Cobb, M A; Bond, R

2011-09-03

The clinical and antibacterial efficacy of two shampoos used as a sole antibacterial treatment in dogs with superficial pyoderma were investigated and compared. In a randomised, partially blinded study, a 3 per cent chlorhexidine gluconate shampoo (Chlorhex 3; Leo Animal Health) was compared against a 2.5 per cent benzoyl peroxide shampoo (Paxcutol; Virbac) in 22 dogs with superficial pyoderma. Dogs were washed two to three times weekly with a 10-minute contact time over 21 days. Clinical scores and bacterial counts were assessed on days 1, 8 and 22 and compared within and between treatment groups; overall response was assessed at the end of the study. Twenty dogs completed the study; 15 (68.2 per cent) showed an overall clinical improvement and the clinical signs resolved in three chlorhexidine-treated dogs. In the chlorhexidine-treated group, scores for papules/pustules (P<0.001), investigator-assessed pruritus (P=0.003), total bacterial counts (P=0.003) and counts for coagulase-positive staphylococci (P=0.003) were reduced after three weeks. Scores and bacterial counts did not vary significantly in the benzoyl peroxide-treated group.

Aerosol, a health hazard during ultrasonic scaling: A clinico-microbiological study.

PubMed

Singh, Akanksha; Shiva Manjunath, R G; Singla, Deepak; Bhattacharya, Hirak S; Sarkar, Arijit; Chandra, Neeraj

2016-01-01

Ultrasonic scaling is a routinely used treatment to remove plaque and calculus from tooth surfaces. These scalers use water as a coolant which is splattered during the vibration of the tip. The splatter when mixed with saliva and plaque of the patients causes the aerosol highly infectious and acts as a major risk factor for transmission of the disease. In spite of necessary protection, sometimes, the operator might get infected because of the infectious nature of the splatter. To evaluate the aerosol contamination produced during ultrasonic scaling by the help of microbiological analysis. This clinico-microbiological study consisted of twenty patients. Two agar plates were used for each patient; the first was kept at the center of the operatory room 20 min before the treatment while the second agar plate was kept 40 cm away from the patient's chest during the treatment. Both the agar plates were sent for microbiological analysis. The statistical analysis was done with the help of STATA 11.0 (StataCorp. 2013. Stata Statistical Software, Release 13. College Station, TX: StataCorp LP, 4905 Lakeway Drive College Station, Texas, USA). Statistical software was used for data analysis and the P < 0.001 was considered to be statistically significant. The results for bacterial count were highly significant when compared before and during the treatment. The Gram staining showed the presence of Staphylococcus and Streptococcus species in high numbers. The aerosols and splatters produced during dental procedures have the potential to spread infection to dental personnel. Therefore, proper precautions should be taken to minimize the risk of infection to the operator.

Validation of the bacterial meningitis score in adults presenting to the ED with meningitis.

PubMed

McArthur, Robert; Edlow, Jonathan A; Nigrovic, Lise E

2016-07-01

The Bacterial Meningitis Score classifies children with meningitis and none of the following high-risk predictors at very low risk for bacterial meningitis: positive cerebrospinal fluid (CSF) Gram stain, CSF protein ≥80mg/dL, CSF absolute neutrophil count (ANC) ≥1000 cells/mm(3), peripheral ANC ≥10,000 cells/mm(3), and seizure at or prior to presentation. Although extensively validated in children, the Bacterial Meningitis Score has not been rigorously evaluated in adults. We performed a single-center cross-sectional retrospective study of adults presenting to the emergency department between 2003 and 2013 with meningitis (defined by CSF white blood cell count ≥10 cells/mm(3)). We defined a case of bacterial meningitis with either a positive CSF or blood culture. We report the performance of the Bacterial Meningitis Score in the study population. We identified 441 eligible patients of which, 4 (1%) had bacterial meningitis. The Bacterial Meningitis Score had a sensitivity of 100% [95% confidence interval (CI) 40%-100%], specificity 51% (95% CI, 46%-56%) and negative predictive value of 100% (95% CI, 98%-100%). None of the low risk adults had bacterial meningitis. If Bacterial Meningitis Score had been applied prospectively, the hospital admission rate would have dropped from 84% to 49% without missing any patients with bacterial meningitis. The Bacterial Meningitis Score accurately identified patients at low risk for bacterial meningitis and could assist clinical decision-making for adults with meningitis. Copyright © 2016 Elsevier Inc. All rights reserved.

A direct plating method for estimating populations of Escherichia coli O157 in bovine manure and manure-based materials.

PubMed

Berry, Elaine D; Wells, James E

2008-11-01

Escherichia coli O157:H7 outbreaks associated with produce consumption have brought attention to livestock manures and manure-based soil amendments as potential sources of pathogens for the contamination of these crops. Procedures for enumeration of E. coli O157:H7 are needed to assess the risks of transmission from these manures and their by-products. A direct plating method employing spiral plating onto CHROMagar O157 was investigated for enumeration of E. coli O157:H7 in feedlot surface material, aged bovine manure, bovine manure compost, and manure-amended soil. In studies utilizing samples spiked with a five-strain cocktail of E. coli O157:H7 at levels ranging from 102 to 10(5) CFU/g of sample, there were strong correlations between the observed and predicted levels of this pathogen. Although the addition of 2.5 mg/liter potassium tellurite and 5 mg/liter novobiocin made the medium more restrictive, these amendments enhanced the ability to identify and enumerate E. coli O157:H7 in feedlot surface material, which contained a higher proportion of fresh feces than did the other three sample types and therefore higher levels of interfering bacterial microflora. The spiral plating method was further assessed to determine its ability to enumerate E. coli O157:H7 in naturally contaminated feedlot surface material. Comparison of E. coli O157:H7 counts in feedlot surface material obtained by the spiral plating method and a most probable number technique were well correlated. We conclude that direct spiral plating onto CHROMagar O157 is effective for estimating E. coli O157:H7 levels in a variety of manures and manure-containing sample types to a lower detection limit of 200 CFU/g. The method has application for determining E. coli O157:H7 concentrations in manures and composts before their sale and use as soil amendments and for measuring the effectiveness of manure treatment processes to reduce or inactivate this pathogen.

Lack of effect of lactose digestion status on baseline fecal microflora

PubMed Central

Szilagyi, Andrew; Shrier, Ian; Chong, George; Je, Jung Sung; Park, Sunghoon; Heilpern, Debra; Lalonde, Catherine; Cote, Louis-Francois; Lee, Byong

2009-01-01

BACKGROUND: The genetics of intestinal lactase divide the world’s population into two phenotypes: the ability (a dominant trait) or inability (a recessive trait) to digest lactose. A prebiotic effect of lactose may impact the colonic flora of these phenotypes differently. OBJECTIVE: To detect and evaluate the effects of lactose on subjects divided according to their ability to digest lactose. METHODS: A total of 57 healthy maldigesters (n=30) and digesters (n=27) completed diet questionnaires, genetic and breath hydrogen testing, and quantitative stool analysis for species of bacteria. Log10 transformation of bacterial counts was compared with lactose intake in both groups using multiple regression analysis. RESULTS: There was a significant relationship between genetic and breath hydrogen tests. Daily lactose intake was marginally lower in lactose maldigesters (median [interquartile range] 12.2 g [31 g] versus 15 g [29.6 g], respectively). There was no relationship between lactose intake and breath hydrogen tests in either group. There were no differences in bacterial counts between the two groups, nor was there a relationship between bacterial counts and lactose intake in either group. CONCLUSION: The differential bacterial effects of lactose were not quantitatively detected in stool samples taken in the present study. PMID:19893771

Effects of storage temperature on bacterial growth rates and community structure in fresh retail sushi.

PubMed

Hoel, S; Jakobsen, A N; Vadstein, O

2017-09-01

This study was conducted to assess the effects of different storage temperatures (4-20°C), on bacterial concentrations, growth rates and community structure in fresh retail sushi, a popular retail product with a claimed shelf life of 2-3 days. The maximum specific growth rate based on aerobic plate count (APC) at 4°C was 0·06 h -1 and displayed a sixfold increase (0·37 h -1 ) at 20°C. Refrigeration resulted in no growth of hydrogen sulphide (H 2 S)-producing bacteria, but this group had the strongest temperature response. The bacterial community structure was determined by PCR/DGGE (denaturing gradient gel electrophoresis). Multivariate analysis based on Bray-Curtis similarities demonstrated that temperature alone was not the major determinant for the bacterial community structure. The total concentration of aerobic bacteria was the variable that most successfully explained the differences between the communities. The dominating organisms, detected by sequencing of DNA bands excised from the DGGE gel, were Brochothrix thermosphacta and genera of lactic acid bacteria (LAB). The relationship between growth rates and storage temperatures clearly demonstrates that these products are sensitive to deviations from optimal storage temperature, possibly resulting in loss of quality during shelf life. Regardless of the storage temperature, the bacterial communities converged towards a similar structure and density, but the storage temperature determined how fast the community reached its carrying capacity. Little information is available on the microbial composition of ready-to-eat food that are prepared with raw fish, subjected to contamination during handling, and susceptible to microbial growth during cold storage. Moreover, the data are a good first possibility to simulate growth of APC, H 2 S-producing bacteria and LAB under different temperature scenarios that might occur during production, distribution or storage. © 2017 The Society for Applied Microbiology.

Bacterial aerosol emission rates from municipal wastewater aeration tanks.

PubMed Central

Sawyer, B; Elenbogen, G; Rao, K C; O'Brien, P; Zenz, D R; Lue-Hing, C

1993-01-01

In this report we describe the results of a study conducted to determine the rates of bacterial aerosol emission from the surfaces of the aeration tanks of the Metropolitan Water Reclamation District of Greater Chicago John E. Egan Water Reclamation Plant. This study was accomplished by conducting test runs in which Andersen six-stage viable samplers were used to collect bacterial aerosol samples inside a walled tower positioned above an aeration tank liquid surface at the John E. Egan Water Reclamation Plant. The samples were analyzed for standard plate counts (SPC), total coliforms (TC), fecal coliforms, and fecal streptococci. Two methods of calculation were used to estimate the bacterial emission rate. The first method was a conventional stack emission rate calculation method in which the measured air concentration of bacteria was multiplied by the air flow rate emanating from the aeration tanks. The second method was a more empirical method in which an attempt was made to measure all of the bacteria emanating from an isolated area (0.37 m2) of the aeration tank surface over time. The data from six test runs were used to determine bacterial emission rates by both calculation methods. As determined by the conventional calculation method, the average SPC emission rate was 1.61 SPC/m2/s (range, 0.66 to 2.65 SPC/m2/s). As determined by the empirical calculation method, the average SPC emission rate was 2.18 SPC/m2/s (range, 1.25 to 2.66 SPC/m2/s). For TC, the average emission rate was 0.20 TC/m2/s (range, 0.02 to 0.40 TC/m2/s) when the conventional calculation method was used and 0.27 TC/m2/s (range, 0.04 to 0.53 TC/m2/s) when the empirical calculation method was used.(ABSTRACT TRUNCATED AT 250 WORDS) PMID:8250547

[Analysis of bactericidal material generated by electrical devices advertising bactericidal ability against bacteria on the agar gel plates].

PubMed

Nishimura, Hidekazu

2012-11-01

Several Japanese companies sell electrical devices advertised as effective in inactivating viruses and killing bacteria by releasing special materials, e.g., Plasmacluster ions, Nanoe particle and minus ions, into the air. These companies claim that their devices killed bacteria on plates in their own experiments. We tested device effectiveness using the same experiments from the Plasmacluster ioniser SHARP Co., Japan, the Nanoe generator Panasonic Co., Japan, and the Vion KING JIM Co., Japan, to test their advertising claims. Bactericidal ability on agar plate was tested, using Staphylococcus aureus, Pseudomonas aeruginosa, Bacillus cereus, and Enterococcus faecalis as follows: the medium containing a certain amount of each bacterium was put onto an agar plate and smeared. Plates were kept in a closed chamber (inner volume 14.4 m3) or a glove box (inner volume 0.2 m), with one of the devices run for 2 hours. Plates not exposed to any device were used as controls. Each plate was retrieved and put in an incubator to count the number of bacterial colonies formed on the plate. There was no significant difference in the number of colonies on plates exposed to devices compared to control, in the number for all devices, or in all bacteria tested in experiments in the 14.4 m3 chamber. These results strongly suggest that these devices have almost no bactericidal effect, at least in space exceeding this volume. Colony formation was suppressed in the glove box in all devices and in all bacteria tested except P. aeruginosa, although the degree of suppression differed among experiments. The colony formation suppression mechanism was analyzed, and indicated that:colony formation did not change even after the removal of Plasmacluster ions, Nanoe particles, or negative ions from the air, while colony formation was decreased drastically by the removal of ozone from space, which was revealed to be generated inevitably during device operation. These results strongly suggest that the bactericidal effect seen only on the agar plate in narrow space was explained by ozone released in space as a by-product, not by special materials as advertising claimed. It is thus important to analyze the effect of special materials such as those done in this study and to suggest the involvement of ozone as the true cause, as have been done in this study, in evaluating bactericidal effect or viral inactivation as advertised by these companies.

Physical impaction injury effects on bacterial cells during spread plating influenced by cell characteristics of the organisms.

PubMed

Thomas, P; Mujawar, M M; Sekhar, A C; Upreti, R

2014-04-01

To understand the factors that contribute to the variations in colony-forming units (CFU) in different bacteria during spread plating. Employing a mix culture of vegetative cells of ten organisms varying in cell characteristics (Gram reaction, cell shape and cell size), spread plating to the extent of just drying the agar surface (50-60 s) was tested in comparison with the alternate spotting-and-tilt-spreading (SATS) approach where 100 μl inoculum was distributed by mere tilting of plate after spotting as 20-25 microdrops. The former imparted a significant reduction in CFU by 20% over the spreader-independent SATS approach. Extending the testing to single organisms, Gram-negative proteobacteria with relatively larger cells (Escherichia, Enterobacter, Agrobacterium, Ralstonia, Pantoea, Pseudomonas and Sphingomonas spp.) showed significant CFU reduction with spread plating except for slow-growing Methylobacterium sp., while those with small rods (Xenophilus sp.) and cocci (Acinetobacter sp.) were less affected. Among Gram-positive nonspore formers, Staphylococcus epidermidis showed significant CFU reduction while Staphylococcus haemolyticus and actinobacteria (Microbacterium, Cellulosimicrobium and Brachybacterium spp.) with small rods/cocci were unaffected. Vegetative cells of Bacillus pumilus and B. subtilis were generally unaffected while others with larger rods (B. thuringiensis, Brevibacillus, Lysinibacillus and Paenibacillus spp.) were significantly affected. A simulated plating study coupled with live-dead bacterial staining endorsed the chances of cell disruption with spreader impaction in afflicted organisms. Significant reduction in CFU could occur during spread plating due to physical impaction injury to bacterial cells depending on the spreader usage and the variable effects on different organisms are determined by Gram reaction, cell size and cell shape. The inoculum spreader could impart physical disruption of vegetative cells against a hard surface. Possibility of CFU reduction in sensitive organisms and the skewed selection of hardier organisms during spread plating, and the recommendation of SATS as an easier and safer alternative for CFU enumerations. © 2013 The Society for Applied Microbiology.

Alternative Hand Contamination Technique To Compare the Activities of Antimicrobial and Nonantimicrobial Soaps under Different Test Conditions▿

PubMed Central

Fuls, Janice L.; Rodgers, Nancy D.; Fischler, George E.; Howard, Jeanne M.; Patel, Monica; Weidner, Patrick L.; Duran, Melani H.

2008-01-01

Antimicrobial hand soaps provide a greater bacterial reduction than nonantimicrobial soaps. However, the link between greater bacterial reduction and a reduction of disease has not been definitively demonstrated. Confounding factors, such as compliance, soap volume, and wash time, may all influence the outcomes of studies. The aim of this work was to examine the effects of wash time and soap volume on the relative activities and the subsequent transfer of bacteria to inanimate objects for antimicrobial and nonantimicrobial soaps. Increasing the wash time from 15 to 30 seconds increased reduction of Shigella flexneri from 2.90 to 3.33 log10 counts (P = 0.086) for the antimicrobial soap, while nonantimicrobial soap achieved reductions of 1.72 and 1.67 log10 counts (P > 0.6). Increasing soap volume increased bacterial reductions for both the antimicrobial and the nonantimicrobial soaps. When the soap volume was normalized based on weight (∼3 g), nonantimicrobial soap reduced Serratia marcescens by 1.08 log10 counts, compared to the 3.83-log10 reduction caused by the antimicrobial soap (P < 0.001). The transfer of Escherichia coli to plastic balls following a 15-second hand wash with antimicrobial soap resulted in a bacterial recovery of 2.49 log10 counts, compared to the 4.22-log10 (P < 0.001) bacterial recovery on balls handled by hands washed with nonantimicrobial soap. This indicates that nonantimicrobial soap was less active and that the effectiveness of antimicrobial soaps can be improved with longer wash time and greater soap volume. The transfer of bacteria to objects was significantly reduced due to greater reduction in bacteria following the use of antimicrobial soap. PMID:18441107

Alternative hand contamination technique to compare the activities of antimicrobial and nonantimicrobial soaps under different test conditions.

PubMed

Fuls, Janice L; Rodgers, Nancy D; Fischler, George E; Howard, Jeanne M; Patel, Monica; Weidner, Patrick L; Duran, Melani H

2008-06-01

Antimicrobial hand soaps provide a greater bacterial reduction than nonantimicrobial soaps. However, the link between greater bacterial reduction and a reduction of disease has not been definitively demonstrated. Confounding factors, such as compliance, soap volume, and wash time, may all influence the outcomes of studies. The aim of this work was to examine the effects of wash time and soap volume on the relative activities and the subsequent transfer of bacteria to inanimate objects for antimicrobial and nonantimicrobial soaps. Increasing the wash time from 15 to 30 seconds increased reduction of Shigella flexneri from 2.90 to 3.33 log(10) counts (P = 0.086) for the antimicrobial soap, while nonantimicrobial soap achieved reductions of 1.72 and 1.67 log(10) counts (P > 0.6). Increasing soap volume increased bacterial reductions for both the antimicrobial and the nonantimicrobial soaps. When the soap volume was normalized based on weight (approximately 3 g), nonantimicrobial soap reduced Serratia marcescens by 1.08 log(10) counts, compared to the 3.83-log(10) reduction caused by the antimicrobial soap (P < 0.001). The transfer of Escherichia coli to plastic balls following a 15-second hand wash with antimicrobial soap resulted in a bacterial recovery of 2.49 log(10) counts, compared to the 4.22-log(10) (P < 0.001) bacterial recovery on balls handled by hands washed with nonantimicrobial soap. This indicates that nonantimicrobial soap was less active and that the effectiveness of antimicrobial soaps can be improved with longer wash time and greater soap volume. The transfer of bacteria to objects was significantly reduced due to greater reduction in bacteria following the use of antimicrobial soap.

The Immediate and Delayed Post-Debridement Effects on Tissue Bacterial Wound Counts of Hypochlorous Acid Versus Saline Irrigation in Chronic Wounds.

PubMed

Hiebert, John M; Robson, Martin C

2016-01-01

Introduction: Wound debridement is considered essential in chronic wound management. Hypochlorous acid has been shown to be an effective agent in reducing wound bacterial counts in open wounds. Ultrasound-enabled wound debridement is an effective and efficient method of debridement. This study compared ultrasound irrigation with hypochlorous acid versus saline irrigation for wound debridement on pre- and postoperative wounds and determined regrowth of bacteria over 1 week period of time. Finally, the outcome of definitive wound closure of the clinically clean-appearing wounds was recorded. Methods: Seventeen consenting adult patients with chronic open wounds were randomly selected for study. The patients were randomly divided into the hypochlorous acid irrigation or saline irrigation group. All patients provided pre- and postoperative tissue samples for qualitative and quantitative bacteriology. For the time (7 days) between the debridement procedure and the definitive closure procedure, the wounds were dressed with a silver-impregnated dressing and a hydroconductive dressing. Results : Both types of irrigation in the ultrasonic system initially lowered the bacterial counts by 4 to 6 logs. However, by the time of definitive closure, the saline-irrigated wounds had bacterial counts back up to 10 5 whereas the hypochlorous acid-irrigated wounds remained at 10 2 or fewer. More than 80% of patients in the saline group had postoperative closure failure compared with 25% of patients in the hypochlorous acid group. Conclusions: Hypochlorous acid irrigation with ultrasound debridement reduced bacterial growth in chronic open wounds more efficiently than saline alone. Postoperative wound closure outcomes suggest a remarkable reduction in wound complications after wound debridement using hypochlorous acid irrigation with ultrasound versus saline alone.

Efficacy of an ethanol/guar/triclosan/glycerine gel on bacteria and yeast loads in canine pododermatitis: a pilot study.

PubMed

Ortalda, C; Noli, C; Cena, T

2016-04-01

To assess efficacy of a gel compound containing guar, glycerine, triclosan and ethanol (Pawcare®, JOKER Technologies, Kerzers, Switzerland) in decreasing bacterial and yeast loads on the paws of dogs with erythematous, greasy and/or malodorous pododermatitis. In 20 dogs, each with at least two affected paws, semiquantitative Malassezia species counts were performed on 10 oil-immersion fields (range: 0 to 30) from acetate tapes pressed on the palmar/plantar surface of one paw. Half of the area was sampled before and the other half immediately after the application of Pawcare(®) . With a similar procedure, swab samples were collected from the other paw for bacterial culture, identification and evaluation of colony-forming units before and immediately after treatment. Statistical evaluation of pre- and posttreatment counts was performed with the Wilcoxon signed-rank test. Nine dogs were positive for Malassezia species Mean acetate tape preparation counts decreased significantly from 8·78 (±8·03) to 5·668 (±6·65) (P=0·0039) after treatment. Twenty-five bacterial isolates of 11 different species were cultured in 19 dogs. Posttreatment cultures were sterile in 8 dogs that had an initial zero or low number (1 to 2 log counts) of colony-forming units. In cases with a higher pre-treatment number of colony forming units (2 to 6 log counts), there was a significant decrease - by a mean of 1·16 log counts (pre 3·12 ±1·69, post 1·96 ±1·57) (P=0·0002). The findings of the present study support the use of PawCare® gel to decrease bacterial and yeast loads in dogs affected by chronic diseases involving the inter-digital spaces. © 2016 British Small Animal Veterinary Association.

Altered Virome and Bacterial Microbiome in Human Immunodeficiency Virus-Associated Acquired Immunodeficiency Syndrome

PubMed Central

Monaco, Cynthia L.; Gootenberg, David B.; Zhao, Guoyan; Handley, Scott A.; Ghebremichael, Musie S.; Lim, Efrem S.; Lankowski, Alex; Baldridge, Megan T.; Wilen, Craig B.; Flagg, Meaghan; Norman, Jason M.; Keller, Brian C.; Luévano, Jesús Mario; Wang, David; Boum, Yap; Martin, Jeffrey N.; Hunt, Peter W.; Bangsberg, David R.; Siedner, Mark J.; Kwon, Douglas S.; Virgin, Herbert W.

2016-01-01

SUMMARY Human immunodeficiency virus (HIV) infection is associated with increased intestinal translocation of microbial products and enteropathy as well as alterations in gut bacterial communities. However, whether the enteric virome contributes to this infection and resulting immunodeficiency remains unknown. We characterized the enteric virome and bacterial microbiome in a cohort of Ugandan patients, including HIV-uninfected or HIV-infected subjects and those either treated with anti-retroviral therapy (ART) or untreated. Low peripheral CD4 T cell counts were associated with an expansion of enteric adenovirus sequences and this increase was independent of ART treatment. Additionally, the enteric bacterial microbiome of patients with lower CD4 T counts exhibited reduced phylogenetic diversity and richness with specific bacteria showing differential abundance, including increases in Enterobacteriaceae, which have been associated with inflammation. Thus, immunodeficiency in progressive HIV infection is associated with alterations in the enteric virome and bacterial microbiome, which may contribute to AIDS-associated enteropathy and disease progression. PMID:26962942

Altered Virome and Bacterial Microbiome in Human Immunodeficiency Virus-Associated Acquired Immunodeficiency Syndrome.

PubMed

Monaco, Cynthia L; Gootenberg, David B; Zhao, Guoyan; Handley, Scott A; Ghebremichael, Musie S; Lim, Efrem S; Lankowski, Alex; Baldridge, Megan T; Wilen, Craig B; Flagg, Meaghan; Norman, Jason M; Keller, Brian C; Luévano, Jesús Mario; Wang, David; Boum, Yap; Martin, Jeffrey N; Hunt, Peter W; Bangsberg, David R; Siedner, Mark J; Kwon, Douglas S; Virgin, Herbert W

2016-03-09

Human immunodeficiency virus (HIV) infection is associated with increased intestinal translocation of microbial products and enteropathy as well as alterations in gut bacterial communities. However, whether the enteric virome contributes to this infection and resulting immunodeficiency remains unknown. We characterized the enteric virome and bacterial microbiome in a cohort of Ugandan patients, including HIV-uninfected or HIV-infected subjects and those either treated with anti-retroviral therapy (ART) or untreated. Low peripheral CD4 T cell counts were associated with an expansion of enteric adenovirus sequences and this increase was independent of ART treatment. Additionally, the enteric bacterial microbiome of patients with lower CD4 T counts exhibited reduced phylogenetic diversity and richness with specific bacteria showing differential abundance, including increases in Enterobacteriaceae, which have been associated with inflammation. Thus, immunodeficiency in progressive HIV infection is associated with alterations in the enteric virome and bacterial microbiome, which may contribute to AIDS-associated enteropathy and disease progression. Copyright © 2016 Elsevier Inc. All rights reserved.

Bacterial contamination monitor

NASA Technical Reports Server (NTRS)

Rich, E.; Macleod, N. H.

1973-01-01

Economical, simple, and fast method uses apparatus which detects bacteria by photography. Apparatus contains camera, film assembly, calibrated light bulb, opaque plastic plate with built-in reflecting surface and transparent window section, opaque slide, plate with chemical packages, and cover containing roller attached to handle.

Microbiological Analysis in Three Diverse Natural Geothermal Bathing Pools in Iceland

PubMed Central

Thorolfsdottir, Berglind Osk Th.; Marteinsson, Viggo Thor

2013-01-01

Natural thermal bathing pools contain geothermal water that is very popular to bathe in but the water is not sterilized, irradiated or treated in any way. Increasing tourism in Iceland will lead to increasing numbers of bath guests, which can in turn affect the microbial flora in the pools and therefore user safety. Today, there is no legislation that applies to natural geothermal pools in Iceland, as the water is not used for consumption and the pools are not defined as public swimming pools. In this study, we conducted a microbiological analysis on three popular but different natural pools in Iceland, located at Lýsuhóll, Hveravellir and Landmannalaugar. Total bacterial counts were performed by flow cytometry, and with plate count at 22 °C, 37 °C and 50 °C. The presence of viable coliforms, Enterococcus spp. and pseudomonads were investigated by growth experiments on selective media. All samples were screened for noroviruses by real time PCR. The results indicate higher fecal contamination in the geothermal pools where the geothermal water flow was low and bathing guest count was high during the day. The number of cultivated Pseudomonas spp. was high (13,000–40,000 cfu/100 mL) in the natural pools, and several strains were isolated and classified as opportunistic pathogens. Norovirus was not detected in the three pools. DNA was extracted from one-liter samples in each pool and analyzed by partial 16S rRNA gene sequencing. Microbial diversity analysis revealed different microbial communities between the pools and they were primarily composed of alpha-, beta- and gammaproteobacteria. PMID:23493033

Adjunctive Steps for Disinfection of the Mandibular Molar Root Canal System: A Correlative Bacteriologic, Micro-Computed Tomography, and Cryopulverization Approach.

PubMed

Alves, Flávio R F; Andrade-Junior, Carlos V; Marceliano-Alves, Marília F; Pérez, Alejandro R; Rôças, Isabela N; Versiani, Marco A; Sousa-Neto, Manoel D; Provenzano, José C; Siqueira, José F

2016-11-01

This study evaluated the disinfecting ability of chemomechanical preparation with rotary nickel-titanium instruments, followed by 2 distinct adjunctive procedures in the root canals of extracted mandibular molars by means of a correlative analytical approach. Twenty-two extracted mandibular molars were selected and anatomically matched between groups on the basis of micro-computed tomographic analysis. In the first phase of the experiment, root canals were contaminated with Enterococcus faecalis and subjected to chemomechanical preparation with BT RaCe instruments and 2.5% NaOCl irrigation. Then either XP-Endo Finisher instrument or passive ultrasonic irrigation was used to supplement disinfection. Micro-computed tomography was used to show whether the percentage of unprepared areas correlated to bacterial counts. In the second phase, the same teeth were contaminated once again, and the adjunctive procedures were used. Samples from the isthmus area of mesial roots and the apical 5-mm fragment of distal roots were obtained by cryopulverization. Samples taken before and after treatment steps in both phases were evaluated by quantitative polymerase chain reaction and statistically analyzed. In phase 1, preparation in both groups resulted in substantial decrease of bacterial counts (P < .001). The adjunctive approaches led to a further small bacterial reduction, which was significant for XP-Endo Finisher (P < .05). No significant differences were observed between groups for persisting bacterial counts. Correlative analysis revealed no statistically significant relationship between bacterial reduction and the percentage of unprepared areas (P > .05). In phase 2, both methods had significant antibacterial effects in the main canal, but none of them could predictably disinfect the isthmus/recess areas. Both XP-Endo Finisher and passive ultrasonic irrigation exhibited antibacterial effectiveness, but only the former caused a significant reduction in the bacterial counts after chemomechanical preparation. None of them were effective in predictably disinfecting the isthmus/recess areas. Copyright © 2016 American Association of Endodontists. Published by Elsevier Inc. All rights reserved.

Does pregnancy have an impact on the subgingival microbiota?

PubMed

Adriaens, Laurence M; Alessandri, Regina; Spörri, Stefan; Lang, Niklaus P; Persson, G Rutger

2009-01-01

We investigated clinical and subgingival microbiologic changes during pregnancy in 20 consecutive pregnant women > or =18 years not receiving dental care. Bacterial samples from weeks 12, 28, and 36 of pregnancy and at 4 to 6 weeks postpartum were processed for 37 species by checkerboard DNA-DNA hybridization. Clinical periodontal data were collected at week 12 and at 4 to 6 weeks postpartum, and bleeding on probing (BOP) was recorded at sites sampled at the four time points. The mean BOP at week 12 and postpartum was 40.1% +/- 18.2% and 27.4% +/- 12.5%, respectively. The corresponding mean BOP at microbiologic test sites was 15% (week 12) and 21% (postpartum; not statistically significant). Total bacterial counts decreased between week 12 and postpartum (P <0.01). Increased bacterial counts over time were found for Neisseria mucosa (P <0.001). Lower counts (P <0.001) were found for Capnocytophaga ochracea, Capnocytophaga sputigena, Eubacterium saburreum, Fusobacterium nucleatum naviforme, Fusobacterium nucleatum polymorphum, Leptotrichia buccalis, Parvimonas micra (previously Peptostreptococcus micros or Micromonas micros), Prevotella intermedia, Prevotella melaninogenica, Staphylococcus aureus, Streptococcus anginosus, Streptococcus intermedius, Streptococcus mutans, Streptococcus oralis, Streptococcus sanguinis, Selenomonas noxia, and Veillonella parvula. No changes occurred between weeks 12 and 28 of pregnancy. Counts of Aggregatibacter actinomycetemcomitans (previously Actinobacillus actinomycetemcomitans), Porphyromonas gingivalis, Tannerella forsythia (previously T. forsythensis), and Treponema denticola did not change. Counts of P. gingivalis and T. forsythia at week 12 were associated with gingivitis (P <0.001). Subgingival levels of bacteria associated with periodontitis did not change. P. gingivalis and T. forsythia counts were associated with BOP at week 12. A decrease was found in 17 of 37 species from week 12 to postpartum. Only counts of N. mucosa increased.

Semiquantitative determination of mesophilic, aerobic microorganisms in cocoa products using the Soleris NF-TVC method.

PubMed

Montei, Carolyn; McDougal, Susan; Mozola, Mark; Rice, Jennifer

2014-01-01

The Soleris Non-fermenting Total Viable Count method was previously validated for a wide variety of food products, including cocoa powder. A matrix extension study was conducted to validate the method for use with cocoa butter and cocoa liquor. Test samples included naturally contaminated cocoa liquor and cocoa butter inoculated with natural microbial flora derived from cocoa liquor. A probability of detection statistical model was used to compare Soleris results at multiple test thresholds (dilutions) with aerobic plate counts determined using the AOAC Official Method 966.23 dilution plating method. Results of the two methods were not statistically different at any dilution level in any of the three trials conducted. The Soleris method offers the advantage of results within 24 h, compared to the 48 h required by standard dilution plating methods.

Growth of Chlorella vulgaris and associated bacteria in photobioreactors

PubMed Central

Lakaniemi, Aino‐Maija; Intihar, Veera M.; Tuovinen, Olli H.; Puhakka, Jaakko A.

2012-01-01

Summary The aim of this study was to test three flat plate photobioreactor configurations for growth of Chlorella vulgaris under non‐axenic conditions and to characterize and quantify associated bacterial communities. The photobioreactor cultivations were conducted using tap water‐based media to introduce background bacterial population. Growth of algae was monitored over time with three independent methods. Additionally, the quantity and quality of eukaryotes and bacteria were analysed using culture‐independent molecular tools based on denaturing gradient gel electrophoresis (PCR‐DGGE) and quantitative polymerase chain reaction (QPCR). Static mixers used in the flat plate photobioreactors did not generally enhance the growth at the low light intensities used. The maximum biomass concentration and maximum specific growth rate were 1.0 g l−1 and 2.0 day−1 respectively. Bacterial growth as determined by QPCR was associated with the growth of C. vulgaris. Based on PCR‐DGGE, bacteria in the cultures mainly originated from the tap water. Bacterial community profiles were diverse but reproducible in all flat plate cultures. Most prominent bacteria in the C. vulgaris cultures belonged to the class Alphaproteobacteria and especially to the genus Sphingomonas. Analysis of the diversity of non‐photosynthetic microorganisms in algal mass cultures can provide useful information on the public health aspects and unravel community interactions. PMID:21936882

Quality and shelf life evaluation of fermented sausages of buffalo meat with different levels of heart and fat.

PubMed

Ahmad, S; Srivastava, P K

2007-04-01

Investigations were carried to study the effect of heart incorporation (0%, 15% and 20%) and increasing levels of fat (20% and 25%) on physicochemical (pH, moisture content and thiobarbituric acid, TBA number) and microbiological (total plate count and yeast and mold count) quality and shelf life of semi dry sausages of buffalo meat during refrigerated storage (4°C). Different levels of fat significantly (p<0.05) increased the pH of the sausage samples. However different levels of heart incorporation did not significantly (p<0.05) affect pH, moisture content and TBA number of sausage samples. Fresh samples had pH, moisture content and TBA number in the range of 5.15-5.28, 42.4-47.4% and 0.073-0.134 respectively. Refrigerated storage significantly (p<0.05) increased TBA number of control samples while storage did not significantly (p<0.05) increase the TBA number of sodium ascorbate (SA) treated samples. Total plate counts of twelve sausage samples were f under the TFTC (too few to count) limit at the initial stage. Incorporation of different levels of heart and also increasing levels of fat did not significantly (p<0.05) increase the log TPC/g values. Yeast and molds were not detected in twelve samples of semi dry fermented sausages in their fresh condition. Storage revealed that there was a consistent decrease in pH, and moisture content. Refrigerated storage significantly (p<0.05) reduced both pH and moisture contents. TBA number and total plate counts and yeast and mold counts of controls were found to increase significantly (p<0.05) during refrigerated storage. However, in SA treated sausage, only TPC and yeast and mold count significantly (p<0.05) increased during refrigerated storage. Shelf life of the sausages was found to be 60 days under refrigerated storage (4°C).

The impact of dairy cows' bedding material and its microbial content on the quality and safety of milk - A cross sectional study of UK farms.

PubMed

Bradley, Andrew J; Leach, Katharine A; Green, Martin J; Gibbons, Jenny; Ohnstad, Ian C; Black, David H; Payne, Barbara; Prout, Victoria E; Breen, James E

2018-03-23

The introduction of bedding dairy cows on recycled manure solids (RMS) in the UK led to concern by competent authorities that there could be an increased, unacceptable risk to animal and human health. A cross-sectional study was designed to evaluate the microbial content of different bedding materials, when used by dairy cows, and its impact on the microbial content of milk. Data were collected from farms bedding lactating cows on sand (n=41), sawdust (n=44) and RMS (n=40). The mean duration of RMS use prior to sampling was 13months. Total bacterial count, and counts of Streptococcus/Enterococcus spp., Staphylococcus spp., Bacillus cereus, thermophilic, thermoduric and psychrotrophic bacteria were determined in used bedding and milk. Samples were evaluated for the presence/absence of Listeria monocytogenes, Salmonella spp. and Yersinia enterocolitica. Data on milking practices were collected to investigate their potential to reduce microbial transfer from bedding to milk. There were substantial differences in bacterial counts both within and between bedding materials. However, there were no significant differences between bedding groups in counts in milk for any of the organisms studied, and no significant correlations between bacterial load in used bedding and milk. Fore-milking was associated with a reduced total bacterial count in milk. Dipping teats with disinfectant and drying, prior to milking, was associated with lower numbers of Streptococcus/Enterococcus spp. in milk. Disinfecting clusters between milking different cows was associated with a reduction in thermophilic and psychrotrophic counts in milk. This study did not provide evidence that use of RMS bedding increased the risk of presence of Y. enterocolitica, Salmonella spp. or L. monocytogenes in milk. However, the strength of this conclusion should be tempered by the relatively small number of farms on which Y. enterocolitica and Salmonella spp. were isolated. It is concluded that, despite the higher bacterial load of RMS, its use as bedding for lactating dairy cows need not be associated with a higher bacterial load in milk than the use of sand or sawdust. However, this finding must be interpreted in the light of the relatively recent introduction of RMS as a bedding material on the farms studied. Teat preparation provides a control point for the potential transfer of microorganisms from bedding to milk. The detection of zoonotic pathogens in a small proportion of milk samples, independent of bedding type, indicates that pasteurisation of milk prior to human consumption remains an important control measure. Copyright © 2017 Elsevier B.V. All rights reserved.

Optimal Irrigation and Debridement of Infected Joint Implants

PubMed Central

Schwechter, Evan M.; Folk, David; Varshney, Avanish K.; Fries, Bettina C.; Kim, Sun Jin; Hirsh, David M.

2014-01-01

Acute postoperative and acute, late hematogenous prosthetic joint infections have been treated with 1-stage irrigation and debridement with polyethylene exchange. Success rates, however, are highly variable. Reported studies demonstrate that detergents are effective at decreasing bacterial colony counts on orthopedic implants. Our hypothesis is that the combination of a detergent and an antiseptic would be more effective than using a detergent alone to decrease colony counts from a methicillin-resistant Staphylococcus aureus biofilm-coated titanium alloy disk simulating an orthopedic implant. In our study of various agents tested, chlorhexidine gluconate scrub (antiseptic and detergent) was the most effective at decreasing bacterial colony counts both prereincubation and postreincubation of the disks; pulse lavage and scrubbing were not more effective than pulse lavage alone. PMID:21641757

Bacteriology of Dehydrated Space Foods 1

PubMed Central

Powers, Edmund M.; Ay, Carl; El-Bisi, Hamed M.; Rowley, Durwood B.

1971-01-01

The initial bacteriological requirement established in 1964 for space foods by the U.S. Army Natick Laboratories are: a total aerobic plate count (≤ 10,000 per g), a total coliform count (≤ 10 per g), fecal coliforms (negative per gram), fecal streptococci (≤ 20 per g), coagulase-positive staphylococci (negative in 5 g) and salmonellae (negative in 10 g). Of the space foods and prototypes tested during 1968 and 1969, 93% complied with the total aerobic plate count, 98% had less than 1 coliform per g, and 99% were negative for fecal coliforms; 88% complied with the streptococci requirement; 100 and 98% were negative for staphylococci and salmonellae, respectively. Nineteen food samples which did not comply (as indicated parenthetically by actual counts per gram) with the requirements were (i) total aerobic plate count: beef soup and gravy base (18,000), chicken soup and gravy base (57,000), spaghetti with meat sauce (12,100 and 14,000), sugared coffee (> 300,000), chocolate ice cream cubes (20,000), and each of four samples of chocolate candy (12,000 to 61,000); (ii) coliforms: two out of three vanilla milk drinks (16 and 127) and one beef hash bar (14); (iii) fecal coliforms: one sample of chicken soup and gravy base positive; (iv) fecal streptococci: two samples of peanut cubes (40 and 108), coconut cubes (75), chicken soup and gravy base (2,650), beef soup and gravy base (33), and five out of six flavored milk drinks (23 to 300); (v) salmonellae: one each of chicken and beef soup and gravy base were positive. Images PMID:4940878

Reflected scatterometry for noninvasive interrogation of bacterial colonies

USDA-ARS?s Scientific Manuscript database

A phenotyping of bacterial colonies on agar plates using forward-scattering diffraction-pattern analysis provided promising classification of several different bacteria such as Salmonella, Vibrio, Listeria, and E. coli. Since the technique is based on forward-scattering phenomena, light transmittanc...

Microbial succession in a compost-packed biofilter treating benzene-contaminated air.

PubMed

Borin, Sara; Marzorati, Massimo; Brusetti, Lorenzo; Zilli, Mario; Cherif, Hanene; Hassen, Abdennaceur; Converti, Attilio; Sorlini, Claudia; Daffonchio, Daniele

2006-03-01

Air artificially contaminated with increasing concentrations of benzene was treated in a laboratory scale compost-packed biofilter for 240 days with a removal efficiency of 81-100%. The bacterial community in the packing material (PM) at different heights of the biofilter was analysed every 60 days. Bacterial plate counts and ribosomal intergenic spacer analysis (RISA) of the isolated strains showed that the number of cultivable aerobic heterotrophic bacteria and the species diversity increased with benzene availability. Identification of the isolated species and the main bands in denaturing gradient gel electrophoresis (DGGE) profiles from total compost DNA during the treatment revealed that, at a relatively low volumetric benzene load (1.2< or =VBL< or =6.4 g m(-3) (PM) h(-1)), besides low G+C Gram positive bacteria, originally present in the packing compost, bacteroidetes and beta- and gamma-proteobacteria became detectable in the colonising population. At the VBL value (24.8 g m(-3) (PM) h(-1)) ensuring the maximum elimination capacity of the biofilter (20.1 g m(-3) (PM) h(-1)), strains affiliated to the genus Rhodococcus dominated the microflora, followed by beta-proteobacteria comprising the genera Bordetella and Neisseria. Under these conditions, more than 35% of the isolated strains were able to grow on benzene as the sole carbon source. Comparison of DGGE and automated RISA profiles of the total community and isolated strains showed that a complex bacterial succession occurred in the reactor in response to the increasing concentrations of the pollutant and that cultivable bacteria played a major role in benzene degradation under the adopted conditions.

Comparison of candidate materials for a synthetic osteo-odonto keratoprosthesis device.

PubMed

Tan, Xiao Wei; Perera, A Promoda P; Tan, Anna; Tan, Donald; Khor, K A; Beuerman, Roger W; Mehta, Jodhbir S

2011-01-05

Osteo-odonto keratoprosthesis is one of the most successful forms of keratoprosthesis surgery for end-stage corneal and ocular surface disease. There is a lack of detailed comparison studies on the biocompatibilities of different materials used in keratoprosthesis. The aim of this investigation was to compare synthetic bioinert materials used for keratoprosthesis surgery with hydroxyapatite (HA) as a reference. Test materials were sintered titanium oxide (TiO(2)), aluminum oxide (Al(2)O(3)), and yttria-stabilized zirconia (YSZ) with density >95%. Bacterial adhesion on the substrates was evaluated using scanning electron microscopy and the spread plate method. Surface properties of the implant discs were scanned using optical microscopy. Human keratocyte attachment and proliferation rates were assessed by cell counting and MTT assay at different time points. Morphologic analysis and immunoblotting were used to evaluate focal adhesion formation, whereas cell adhesion force was measured with a multimode atomic force microscope. The authors found that bacterial adhesion on the TiO(2), Al(2)O(3), and YSZ surfaces were lower than that on HA substrates. TiO(2) significantly promoted keratocyte proliferation and viability compared with HA, Al(2)O(3,) and YSZ. Immunofluorescent imaging analyses, immunoblotting, and atomic force microscope measurement revealed that TiO(2) surfaces enhanced cell spreading and cell adhesion compared with HA and Al(2)O(3). TiO(2) is the most suitable replacement candidate for use as skirt material because it enhanced cell functions and reduced bacterial adhesion. This would, in turn, enhance tissue integration and reduce device failure rates during keratoprosthesis surgery.

Cultivable anaerobic and aerobic bacterial communities in the fermentation chambers of Holotrichia parallela (coleoptera: scarabaeidae) larvae.

PubMed

Zhang, Zhen-Yu; Yuan, Yimin; Ali, Muhammad Waqar; Peng, Tao; Peng, Wei; Raza, Muhammad Fahim; Zhao, Yongshun; Zhang, Hongyu

2018-01-01

As important pests, scarab beetle larvae survive on plant biomass and the microbiota of the fermentation chamber play an important role in the digestion of lignocellulose-rich diets. However, the cultivable microbes, especially the anaerobic cultivable microbes, are still largely unknown. Here, both cultivable anaerobic and aerobic bacterial communities associated with the fermentation chamber of Holotrichia parallela larvae were investigated. In total bacteria cells directly enumerated by the 4', 6-diamidino-2-phenylindole (DAPI) staining method, the viable plate counts of cultivable bacteria in the fermentation chamber accounted for 0.92% of proportion. These cultivable bacteria were prone to attach to the fermentation chamber wall (88.41%) compared to the chamber contents. Anaerobic bacteria were dominant in the cultivable bacteria attaching to the fermentation chamber wall (70.20%), while the quantities of anaerobes and aerobes were similar in the chamber contents. Polymerase chain reaction-denaturing gradient gel electrophoresis (PCR-DGGE), fingerprinting and sequence analysis of isolated colonies revealed that the cultivable bacteria are affiliated with class γ-Proteobacteria, Bacteroidia, Actinobacteria, Clostridia and β-Proteobacteria. γ-Proteobacteria was the major type of anaerobic cultivable bacteria and even the only one type of aerobic cultivable bacteria. Taken together, our results suggest, for the first time, that anaerobic microbiota are dominant in cultivable bacteria in the special anoxia niche of the fermentation chamber from H. parallela larvae. These bacterial isolates could be a treasure trove for screening lignocellulytic microbes which are essential for the plant biomass digestion of this scarab species.

Cultivable anaerobic and aerobic bacterial communities in the fermentation chambers of Holotrichia parallela (coleoptera: scarabaeidae) larvae

PubMed Central

Ali, Muhammad Waqar; Peng, Tao; Peng, Wei; Raza, Muhammad Fahim; Zhao, Yongshun; Zhang, Hongyu

2018-01-01

As important pests, scarab beetle larvae survive on plant biomass and the microbiota of the fermentation chamber play an important role in the digestion of lignocellulose-rich diets. However, the cultivable microbes, especially the anaerobic cultivable microbes, are still largely unknown. Here, both cultivable anaerobic and aerobic bacterial communities associated with the fermentation chamber of Holotrichia parallela larvae were investigated. In total bacteria cells directly enumerated by the 4’, 6-diamidino-2-phenylindole (DAPI) staining method, the viable plate counts of cultivable bacteria in the fermentation chamber accounted for 0.92% of proportion. These cultivable bacteria were prone to attach to the fermentation chamber wall (88.41%) compared to the chamber contents. Anaerobic bacteria were dominant in the cultivable bacteria attaching to the fermentation chamber wall (70.20%), while the quantities of anaerobes and aerobes were similar in the chamber contents. Polymerase chain reaction-denaturing gradient gel electrophoresis (PCR-DGGE), fingerprinting and sequence analysis of isolated colonies revealed that the cultivable bacteria are affiliated with class γ-Proteobacteria, Bacteroidia, Actinobacteria, Clostridia and β-Proteobacteria. γ-Proteobacteria was the major type of anaerobic cultivable bacteria and even the only one type of aerobic cultivable bacteria. Taken together, our results suggest, for the first time, that anaerobic microbiota are dominant in cultivable bacteria in the special anoxia niche of the fermentation chamber from H. parallela larvae. These bacterial isolates could be a treasure trove for screening lignocellulytic microbes which are essential for the plant biomass digestion of this scarab species. PMID:29304141

Temporal variations in the abundance and composition of biofilm communities colonizing drinking water distribution pipes.

PubMed

Kelly, John J; Minalt, Nicole; Culotti, Alessandro; Pryor, Marsha; Packman, Aaron

2014-01-01

Pipes that transport drinking water through municipal drinking water distribution systems (DWDS) are challenging habitats for microorganisms. Distribution networks are dark, oligotrophic and contain disinfectants; yet microbes frequently form biofilms attached to interior surfaces of DWDS pipes. Relatively little is known about the species composition and ecology of these biofilms due to challenges associated with sample acquisition from actual DWDS. We report the analysis of biofilms from five pipe samples collected from the same region of a DWDS in Florida, USA, over an 18 month period between February 2011 and August 2012. The bacterial abundance and composition of biofilm communities within the pipes were analyzed by heterotrophic plate counts and tag pyrosequencing of 16S rRNA genes, respectively. Bacterial numbers varied significantly based on sampling date and were positively correlated with water temperature and the concentration of nitrate. However, there was no significant relationship between the concentration of disinfectant in the drinking water (monochloramine) and the abundance of bacteria within the biofilms. Pyrosequencing analysis identified a total of 677 operational taxonomic units (OTUs) (3% distance) within the biofilms but indicated that community diversity was low and varied between sampling dates. Biofilms were dominated by a few taxa, specifically Methylomonas, Acinetobacter, Mycobacterium, and Xanthomonadaceae, and the dominant taxa within the biofilms varied dramatically between sampling times. The drinking water characteristics most strongly correlated with bacterial community composition were concentrations of nitrate, ammonium, total chlorine and monochloramine, as well as alkalinity and hardness. Biofilms from the sampling date with the highest nitrate concentration were the most abundant and diverse and were dominated by Acinetobacter.

Formation and resuscitation of viable but nonculturable Salmonella typhi.

PubMed

Zeng, Bin; Zhao, Guozhong; Cao, Xiaohong; Yang, Zhen; Wang, Chunling; Hou, Lihua

2013-01-01

Salmonella typhi is a pathogen that causes the human disease of typhoid fever. The aim of this study was to investigate the viable but nonculturable (VBNC) state of S. typhi. Some samples were stimulated at 4°C or -20°C, while others were induced by different concentrations of CuSO4. Total cell counts remained constant throughout several days by acridine orange direct counting; however, plate counts declined to undetectable levels within 48 hours by plate counting at -20°C. The direct viable counts remained fairly constant at this level by direct viable counting. Carbon and nitrogen materials slowly decreased which indicated that a large population of cells existed in the VBNC state and entered the VBNC state in response to exposure to 0.01 or 0.015 mmol/L CuSO4 for more than 14 or 12 days, respectively. Adding 3% Tween 20 or 1% catalase enabled cells to become culturable again, with resuscitation times of 48 h and 24 h, respectively. The atomic force microscope results showed that cells gradually changed in shape from short rods to coccoids, and decreased in size when they entered the VBNC state. Further animal experiments suggested that resuscitated cells might regain pathogenicity.

A Sortase A-Immobilized Mesoporous Hollow Carbon Sphere-Based Biosensor for Detection of Gram-Positive Bacteria

NASA Astrophysics Data System (ADS)

Wang, Hongsu; Luo, Ruiping; Chen, Yang; Si, Qi; Niu, Xiaodi

2018-05-01

A sensor based on mesoporous carbon materials immobilized with sortase A (SrtA) for determination of Staphylococcus aureus (S. aureus) is reported. To prepare the biosensor, we first synthesized carboxyl-functionalized mesoporous hollow carbon spheres, then applied them as carriers for immobilization of SrtA. Based on the catalytic mechanism of SrtA, a highly sensitive, inexpensive, and rapid method was developed for S. aureus detection. The sensor showed a linear response in the bacterial concentration range of 0.125 × 102 colony-forming units (CFU) mL-1 to 2.5 × 102 CFU mL-1, with detection limit as low as 9.0 CFU mL-1. The method was successfully used for quantitative detection of S. aureus in whole milk samples, giving results similar to experimental results obtained from the plate counting method. This biosensor could also be used to detect other Gram-positive bacteria that secrete SrtA.

Cell viability of mycorrhiza helper bacteria solid inoculant in different carrier material

NASA Astrophysics Data System (ADS)

Asyiah, Iis Nur; Hindersah, Reginawanti; Harni, Rita

2018-02-01

Roots of food crops are colonized by nonpathogenic mycorrhizal fungi which show natural ability to control plant pathogen. Mycorrhizal establishment in plant roots is affected by rhizobacteria, known as mycorrhiza helper bacteria (MHB), which has synergetic effects on mycorrhizal associations. Laboratory experiment has been conducted to assess the best carrier material to develop well-qualified MHB of Pseudomonas diminuta and Bacillus subtilis solid inoculant. Carrier materials were 100 mesh organic matter of agricultural waste. Different spore concentration of both bacterial liquid inoculants were grown on three kinds of 100-mesh organic matter and stored at room temperature up to 90 days. Cell viability of both MHB were counted by serial dilution plate method by using specific medium. The results showed that sugar cane baggase ash was the best carrier material to maintain cell viability for both MHB. However, the population of Pseudomonas diminuta and Bacillus subtilis in sugar cane baggase ash were slightly decreased after 90 days. The use of sugarcane baggase ash for solid MHB inoculant development could be suggested.

The Mediterranean non-indigenous ascidian Polyandrocarpa zorritensis: Microbiological accumulation capability and environmental implications.

PubMed

Stabili, Loredana; Licciano, Margherita; Longo, Caterina; Lezzi, Marco; Giangrande, Adriana

2015-12-15

We investigated the bacterial accumulation and digestion capability of Polyandrocarpa zorritensis, a non-indigenous colonial ascidian originally described in Peru and later found in the Mediterranean. Microbiological analyses were carried out on homogenates from "unstarved" and "starved" ascidians and seawater from the same sampling site (Adriatic Sea, Italy). Culturable heterotrophic bacteria (22 °C), total culturable bacteria (37 °C) and vibrios abundances were determined on Marine Agar 2216, Plate Count Agar and TCBS Agar, respectively. Microbial pollution indicators were measured by the most probable number method. All the examined microbiological groups were accumulated by ascidians but differently digested. An interesting outcome is the capability of P. zorritensis to digest allochthonous microorganisms such as coliforms as well as culturable bacteria at 37 °C, counteracting the effects of microbial pollution. Thus, the potential exploitation of these filter feeders to restore polluted seawater should be taken into consideration in the management of this alien species. Copyright © 2015 Elsevier Ltd. All rights reserved.

Decreased mortality of weaned pigs with Streptococcus suis with the use of in-water potassium penicillin G.

PubMed

Byra, Chris; Gadbois, Pierre; Cox, William R; Gottschalk, Marcelo; Farzan, Vahab; Bauer, Sharon A; Wilson, Jeff B

2011-03-01

This study evaluated the efficacy of potassium penicillin G in drinking water of weaned pigs to reduce mortality and spread of infection caused by Streptococcus suis. A total of 896 18-day-old weaned pigs were randomly assigned to either treatment with potassium penicillin G in-water (Treated), or no treatment (Control). The outcomes analyzed were total mortality, mortality due to S. suis, and overall counts of S. suis colonies. The risk of mortality due to S. suis and total mortality were significantly increased in the Control group compared with Treated pigs (P < 0.05). Bacterial culture of posterior pharyngeal swabs indicated that Control pigs were significantly more likely to have ≥ 1000 colonies of S. suis per plate than were Treated pigs (P < 0.05). This study demonstrates that potassium penicillin G administered in drinking water is effective in reducing mortality associated with S. suis infection and reducing tonsillar carriage of S. suis.

Decreased mortality of weaned pigs with Streptococcus suis with the use of in-water potassium penicillin G

PubMed Central

Byra, Chris; Gadbois, Pierre; Cox, William R.; Gottschalk, Marcelo; Farzan, Vahab; Bauer, Sharon A.; Wilson, Jeff B.

2011-01-01

This study evaluated the efficacy of potassium penicillin G in drinking water of weaned pigs to reduce mortality and spread of infection caused by Streptococcus suis. A total of 896 18-day-old weaned pigs were randomly assigned to either treatment with potassium penicillin G in-water (Treated), or no treatment (Control). The outcomes analyzed were total mortality, mortality due to S. suis, and overall counts of S. suis colonies. The risk of mortality due to S. suis and total mortality were significantly increased in the Control group compared with Treated pigs (P < 0.05). Bacterial culture of posterior pharyngeal swabs indicated that Control pigs were significantly more likely to have ≥ 1000 colonies of S. suis per plate than were Treated pigs (P < 0.05). This study demonstrates that potassium penicillin G administered in drinking water is effective in reducing mortality associated with S. suis infection and reducing tonsillar carriage of S. suis. PMID:21629419

Effects of combined sewer overflow and stormwater on indicator bacteria concentrations in the Tama River due to the high population density of Tokyo Metropolitan area.

PubMed

Ham, Young-Sik; Kobori, Hiromi; Takasago, Masahisa

2009-05-01

The indicator bacteria (standard plate count, total coliform, and fecal coliform bacteria) concentrations have been investigated using six ambient habitats (population density, percent sewer penetration, stream flow rate (m(3)/sec), percent residential area, percent forest area and percent agricultural area) in the Tama River basin in Tokyo, Japan during June 2003 to January 2005. The downstream and tributary Tama River showed higher concentrations of TC and FC bacteria than the upstream waters, which exceeded an environmental quality standard for rivers and a bathing water quality criterion. It was estimated that combined sewer overflow (CSO) and stormwater effluents contributed -4-23% to the indicator bacteria concentrations of the Tama River. The results of multiple regression analyses show that the indicator bacteria concentrations of Tama River basin are significantly affected by population density. It is concluded that the Tama River received a significant bacterial contamination load originating from the anthropogenic source.