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

Site-specific mouth rinsing can improve oral odor by altering bacterial counts. Blind crossover clinical study.

PubMed

Alqumber, Mohammed A; Arafa, Khaled A

2014-11-01

To determine whether site-specific mouth rinsing with oral disinfectants can improve oral odor beyond the traditional panoral mouth disinfection with mouth rinses by targeting specifically oral malodor implicated anaerobic bacteria. Twenty healthy fasting subjects volunteered for a blinded prospective, descriptive correlational crossover cross-section clinical trial conducted during the month of Ramadan between July and August 2013 in Albaha province in Saudi Arabia involving the application of Listerine Cool Mint mouth rinse by either the traditional panoral rinsing method, or a site-specific disinfection method targeting the subgingival and supragingival plaque and the posterior third of the tongue dorsum, while avoiding the remaining locations within the oral cavity. The viable anaerobic and aerobic bacterial counts, volatile sulfur compounds (VSCs) levels, organoleptic assessment of oral odor, and the tongue-coating index were compared at baseline, one, 5, and 9 hours after the treatment. The site-specific disinfection method reduced the VSCs and anaerobic bacterial loads while keeping the aerobic bacterial numbers higher than the traditional panoral rinsing method. Site-specific disinfection can more effectively maintain a healthy oral cavity by predominantly disinfecting the niches of anaerobic bacteria within the oral cavity.

Distribution and identification of culturable airborne microorganisms in a Swiss milk processing facility.

PubMed

Brandl, Helmut; Fricker-Feer, Claudia; Ziegler, Dominik; Mandal, Jyotshna; Stephan, Roger; Lehner, Angelika

2014-01-01

Airborne communities (mainly bacteria) were sampled and characterized (concentration levels and diversity) at 1 outdoor and 6 indoor sites within a Swiss dairy production facility. Air samples were collected on 2 sampling dates in different seasons, one in February and one in July 2012 using impaction bioaerosol samplers. After cultivation, isolates were identified by mass spectrometry (matrix-assisted laser desorption/ionization-time-of-flight) and molecular (sequencing of 16S rRNA and rpoB genes) methods. In general, total airborne particle loads and total bacterial counts were higher in winter than in summer, but remained constant within each indoor sampling site at both sampling times (February and July). Bacterial numbers were generally very low (<100 cfu/m(3) of air) during the different steps of milk powder production. Elevated bacterial concentrations (with mean values of 391 ± 142 and 179 ± 33 cfu/m(3) of air during winter and summer sampling, respectively; n=15) occurred mainly in the "logistics area," where products in closed tins are packed in secondary packaging material and prepared for shipping. However, total bacterial counts at the outdoor site varied, with a 5- to 6-fold higher concentration observed in winter compared with summer. Twenty-five gram-positive and gram-negative genera were identified as part of the airborne microflora, with Bacillus and Staphylococcus being the most frequent genera identified. Overall, the culturable microflora community showed a composition typical and representative for the specific location. Bacterial counts were highly correlated with total airborne particles in the size range 1 to 5 µm, indicating that a simple surveillance system based upon counting of airborne particles could be implemented. The data generated in this study could be used to evaluate the effectiveness of the dairy plant's sanitation program and to identify potential sources of airborne contamination, resulting in increased food safety. Copyright © 2014 American Dairy Science Association. Published by Elsevier Inc. All rights reserved.

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.

A combination of thalidomide and augmentin protects BALB/c mice suffering from Klebsiella pneumoniae B5055-induced sepsis.

PubMed

Kumar, V; Harjai, K; Chhibber, S

2009-04-01

Despite extensive research, the mortality associated with sepsis in hospitals remains very high. We have evaluated the protective immunomodulatory effect of thalidomide alone or with Augmentin in Klebsiella pneumoniae B5055-induced sepsis in BALB/c mice. The mouse model of sepsis was developed by placing K. pneumoniae B5055 entrapped in fibrin and thrombin clots in the peritoneal cavity of mice. The septic mice were treated with thalidomide alone (30 mg/kg/day/po), Augmentin alone (20 microg/ml/ip) and with their combination. the thalidomide-alone treated mice showed 75% survival whereas 60% of the Augmentin-alone treated group survived. Combination treatment provided 100% survival. Treatment with thalidomide alone significantly (p<0.05) decreased interleukin-1 alpha (IL-1alpha), nitric oxide (NO) and malondialdehyde (MDA) levels in the serum without significantly (p<0.05) decreasing the bacterial count in blood. Augmentin alone only decreased the bacterial load in blood significantly (p<0.05). However, a combination of thalidomide with Augmentin significantly (p<0.05) decreased both the bacterial count and inflammatory mediators.

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.

Short communication: Repeatability of differential goat bulk milk culture and associations with somatic cell count, total bacterial count, and standard plate count.

PubMed

Koop, G; Dik, N; Nielen, M; Lipman, L J A

2010-06-01

The aims of this study were to assess how different bacterial groups in bulk milk are related to bulk milk somatic cell count (SCC), bulk milk total bacterial count (TBC), and bulk milk standard plate count (SPC) and to measure the repeatability of bulk milk culturing. On 53 Dutch dairy goat farms, 3 bulk milk samples were collected at intervals of 2 wk. The samples were cultured for SPC, coliform count, and staphylococcal count and for the presence of Staphylococcus aureus. Furthermore, SCC (Fossomatic 5000, Foss, Hillerød, Denmark) and TBC (BactoScan FC 150, Foss) were measured. Staphylococcal count was correlated to SCC (r=0.40), TBC (r=0.51), and SPC (r=0.53). Coliform count was correlated to TBC (r=0.33), but not to any of the other variables. Staphylococcus aureus did not correlate to SCC. The contribution of the staphylococcal count to the SPC was 31%, whereas the coliform count comprised only 1% of the SPC. The agreement of the repeated measurements was low. This study indicates that staphylococci in goat bulk milk are related to SCC and make a significant contribution to SPC. Because of the high variation in bacterial counts, repeated sampling is necessary to draw valid conclusions from bulk milk culturing. 2010 American Dairy Science Association. Published by Elsevier Inc. All rights reserved.

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.

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.

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

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.

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.

HETEROTROPHIC PLATE COUNT (HPC) METHODOLOGY IN THE UNITED STATES

EPA Science Inventory

ABSTRACT

In the United States (U.S.), the history of bacterial plate counting methods used for water can be traced largely through Standard Methods for the Examination of Water and Wastewater (Standard Methods). The bacterial count method has evolved from the original St...

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.

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

Biofilms associated with poultry processing equipment.

PubMed

Lindsay, D; Geornaras, I; von Holy, A

1996-01-01

Aerobic and Gram-negative bacteria were enumerated on non-metallic surfaces and stainless steel test pieces attached to equipment surfaces by swabbing and a mechanical dislodging procedure, respectively, in a South African grade B poultry processing plant. Changes in bacterial numbers were also monitored over time on metal test pieces. The highest bacterial counts were obtained from non-metallic surfaces such as rubber fingered pluckers and plastic defeathering curtains which exceeded the highest counts found on the metal surfaces by at least 1 log CFU cm-2. Gram-negative bacterial counts on all non-metallic surface types were at least 2 log CFU cm-2 lower than corresponding aerobic plate counts. On metal surfaces, the highest microbial numbers were obtained after 14 days exposure, with aerobic plate counts ranging from 3.57 log CFU cm-2 to 5.13 log CFU cm-2, and Gram-negative counts from 0.70 log CFU cm-2 to 3.31 log CFU cm-2. Scanning electron microscopy confirmed the presence of bacterial cells on non-metallic and metallic surfaces associated with poultry processing. Rubber 'fingers', plastic curtains, conveyor belt material and stainless steel test surfaces placed on the scald tank overflow and several chutes revealed extensive and often confluent bacterial biofilms. Extracellular polymeric substances, but few bacterial cells were visible on test pieces placed on evisceration equipment, spinchiller blades and the spinchiller outlet.

Comparing the Effectiveness of Polymer Debriding Devices Using a Porcine Wound Biofilm Model.

PubMed

Wilkinson, Holly N; McBain, Andrew J; Stephenson, Christian; Hardman, Matthew J

2016-11-01

Objective: Debridement to remove necrotic and/or infected tissue and promote active healing remains a cornerstone of contemporary chronic wound management. While there has been a recent shift toward less invasive polymer-based debriding devices, their efficacy requires rigorous evaluation. Approach: This study was designed to directly compare monofilament debriding devices to traditional gauze using a wounded porcine skin biofilm model with standardized application parameters. Biofilm removal was determined using a surface viability assay, bacterial counts, histological assessment, and scanning electron microscopy (SEM). Results: Quantitative analysis revealed that monofilament debriding devices outperformed the standard gauze, resulting in up to 100-fold greater reduction in bacterial counts. Interestingly, histological and morphological analyses suggested that debridement not only removed bacteria, but also differentially disrupted the bacterially-derived extracellular polymeric substance. Finally, SEM of post-debridement monofilaments showed structural changes in attached bacteria, implying a negative impact on viability. Innovation: This is the first study to combine controlled and defined debridement application with a biologically relevant ex vivo biofilm model to directly compare monofilament debriding devices. Conclusion: These data support the use of monofilament debriding devices for the removal of established wound biofilms and suggest variable efficacy towards biofilms composed of different species of bacteria.

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.

Phenotypic Characterization and Antibiogram of CSF Isolates in Acute Bacterial Meningitis.

PubMed

Modi, Syamal; Anand, Amit Kumar

2013-12-01

Acute bacterial meningitis (ABM) is a medical emergency, which warrants an early diagnosis and an aggressive therapy. Despite the availability of potent newer antibiotics, the mortality rate caused by acute bacterial meningitis remains significantly high in India and in other developing countries, which ranges from 16 - 32%. There is a need of a periodic review of bacterial meningitis worldwide, since the pathogens which are responsible for the infection may vary with time, geography and the age of the patient. Our aim was to study the bacterial profiles and antimicrobial susceptibility patterns of the CSF isolates which were obtained from patients of acute bacterial meningitis in our area. Two hundred and fifty two clinically diagnosed cases of acute bacterial meningitis, who were admitted to the wards of a tertiary medical centre in Patna, during the period from August 2011 to December 2012, were included in this study. Two hundred and fifty two CSF samples from as many patients of ABM were processed for cell counts, biochemical analysis, gram staining, culture, antigen detection by latex agglutination test (LAT) and antibiotic susceptibility tests, as per the standard techniques. In this study, 62.3% patients were males and 37.7% were females The most common age group of presentation was 12-60 years (80.2%). Gram stained smears were positive in 162 (64.3%) samples, while culture yielded positive growth in 200 (79.4%) patients. Streptococcus pneumoniae was the most common pathogen which was isolated in 120 (60%) culture positive cases. Cell counts showed the predominance of neutrophils in all cases with ABM. High protein and low sugar levels correlated well with the features of ABM. All gram positive isolates were sensitive to vancomycin. All the gram negative isolates were sensitive to imipenem. Twenty two (8.7%) patients expired during the course of study. Deaths were caused by N.meningitidis in 9 (40.9%) cases, by S.pneumoniae in 3 (13.6%) cases and by H.influenzea in 1 (4.5%) case. In the remaining 9 (40.9%) mortality cases, the organism could not be identified. Simple, rapid, inexpensive tests like gram staining remain significant means for diagnosis of ABM in developing countries. LAT for pneumococcal antigen should be performed first, since Streptococcus pneumoniae remains the major aetiological agent of ABM, both in adults and children. The final diagnosis of ABM depends upon a comprehensive analysis of CSF smears, cultures, LAT, cytological, biochemical and clinical findings of the cases, and a single test or parameter cannot be used to decide the course of management in the patients. However, empirical therapy is advocated, considering the potentially high rate of mortality in these patients.

Surface Enhanced Raman Spectroscopy for the Rapid Detection and Identification of Microbial Pathogens in Human Serum

DTIC Science & Technology

2014-12-11

and 1 mm depth. Bacterial culture and cell count determination Bacterial species of Acinetobacter baumannii (A. baumannii, ST-3), Escherichia coli...remove all broth components followed by a final resuspension of the pellet in ddH2O back to 1 OD. Cell count was determined by plating the 10 4 , 10 3...10 2 and 10 1 cell dilutions on TSB Nutrient Agar media. Colony forming units (CFU) were counted the following day to confirm bacterial species

Automatic counting and classification of bacterial colonies using hyperspectral imaging

USDA-ARS?s Scientific Manuscript database

Detection and counting of bacterial colonies on agar plates is a routine microbiology practice to get a rough estimate of the number of viable cells in a sample. There have been a variety of different automatic colony counting systems and software algorithms mainly based on color or gray-scale pictu...

A cocktail of humanized anti-pertussis toxin antibodies limits disease in murine and baboon models of whooping cough.

PubMed

Nguyen, Annalee W; Wagner, Ellen K; Laber, Joshua R; Goodfield, Laura L; Smallridge, William E; Harvill, Eric T; Papin, James F; Wolf, Roman F; Padlan, Eduardo A; Bristol, Andy; Kaleko, Michael; Maynard, Jennifer A

2015-12-02

Despite widespread vaccination, pertussis rates are rising in industrialized countries and remain high worldwide. With no specific therapeutics to treat disease, pertussis continues to cause considerable infant morbidity and mortality. The pertussis toxin is a major contributor to disease, responsible for local and systemic effects including leukocytosis and immunosuppression. We humanized two murine monoclonal antibodies that neutralize pertussis toxin and expressed them as human immunoglobulin G1 molecules with no loss of affinity or in vitro neutralization activity. When administered prophylactically to mice as a binary cocktail, antibody treatment completely mitigated the Bordetella pertussis-induced rise in white blood cell counts and decreased bacterial colonization. When administered therapeutically to baboons, antibody-treated, but not untreated control animals, experienced a blunted rise in white blood cell counts and accelerated bacterial clearance rates. These preliminary findings support further investigation into the use of these antibodies to treat human neonatal pertussis in conjunction with antibiotics and supportive care. Copyright © 2015, American Association for the Advancement of Science.

A cocktail of humanized anti-pertussis toxin antibodies limits disease in murine and baboon models of whooping cough

PubMed Central

Nguyen, Annalee W.; Wagner, Ellen K.; Laber, Joshua R.; Goodfield, Laura L.; Smallridge, William E.; Harvill, Eric T.; Papin, James F.; Wolf, Roman F.; Padlan, Eduardo A.; Bristol, Andy; Kaleko, Michael; Maynard, Jennifer A.

2016-01-01

In spite of wide-spread vaccination, pertussis rates are rising in industrialized countries and remain high world-wide. With no specific therapeutics to treat disease, pertussis continues to cause considerable infant morbidity and mortality. The pertussis toxin is a major contributor to disease, responsible for local and systemic effects including leukocytosis and immunosuppression. Here, we humanized two murine monoclonal antibodies that neutralize pertussis toxin and expressed them as human IgG1 molecules with no loss of affinity or in vitro neutralization activity. When administered prophylactically to mice as a binary cocktail, antibody treatment completely mitigated the B. pertussis-induced rise in white blood cell count and decreased bacterial colonization. When administered therapeutically to baboons, antibody-treated but not control animals experienced a blunted rise in white blood cell count and accelerated bacterial clearance rates. These preliminary findings support further investigation into the use of these antibodies to treat human neonatal pertussis in conjunction with antibiotics and supportive care. PMID:26631634

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.

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.

Survival of Staphylococcus pseudintermedius in modified Romanowsky staining solutions.

PubMed

Misan, Angus; Chan, Wei Yee; Trott, Darren; Hill, Peter B

2017-08-01

Stains that are used regularly for patient-side diagnosis to rapidly identify bacterial and fungal infections could become contaminated by common pathogens, such as Staphylococcus pseudintermedius, during slide immersion. To determine whether the inoculation of S. pseudintermedius into modified Romanowsky type stains (Quick Dip ® ) results in viable bacterial contamination and whether this is influenced by the addition of organic debris (canine hair and skin). A clinical isolate of S. pseudintermedius was inoculated into clean and organically contaminated Quick Dip ® solutions (methanol fixative, eosin, methylene blue), and positive (broth) and negative (bleach) controls. Each solution was tested for the presence of viable bacteria by counting the number of colony forming units (CFU/mL) at various time points. Solutions also were examined under high power microscopy to count the number of visible bacteria at each time point. Staphylococcus pseudintermedius was able to survive in the clean and contaminated Quick Dip ® stains for at least one hour, but by 24 h no viable bacteria remained. Survival of the bacteria was not supported in the fixative at any time point. Staphylococcus pseudintermedius remained visible under high power microscopy for up to 2 weeks in all organically contaminated solutions of the Quick Dip ® set. Staphylococcus pseudintermedius only remains viable in eosin and methylene blue for short periods of time, but the prolonged visibility of dead organisms could theoretically lead to the misdiagnosis of cytology samples. © 2017 ESVD and ACVD.

Microbiological assessment of house and imported bottled water by comparison of bacterial endotoxin concentration, heterotrophic plate count, and fecal coliform count.

PubMed

Reyes, Mayra I; Pérez, Cynthia M; Negrón, Edna L

2008-03-01

Consumers increasingly use bottled water and home water treatment systems to avoid direct tap water. According to the International Bottled Water Association (IBWA), an industry trade group, 5 billion gallons of bottled water were consumed by North Americans in 2001. The principal aim of this study was to assess the microbial quality of in-house and imported bottled water for human consumption, by measurement and comparison of the concentration of bacterial endotoxin and standard cultivable methods of indicator microorganisms, specifically, heterotrophic and fecal coliform plate counts. A total of 21 brands of commercial bottled water, consisting of 10 imported and 11 in-house brands, selected at random from 96 brands that are consumed in Puerto Rico, were tested at three different time intervals. The Standard Limulus Amebocyte Lysate test, gel clot method, was used to measure the endotoxin concentrations. The minimum endotoxin concentration in 63 water samples was less than 0.0625 EU/mL, while the maximum was 32 EU/mL. The minimum bacterial count showed no growth, while the maximum was 7,500 CFU/mL. Bacterial isolates like P. fluorescens, Corynebacterium sp. J-K, S. paucimobilis, P. versicularis, A. baumannii, P. chlororaphis, F. indologenes, A. faecalis and P. cepacia were identified. Repeated measures analysis of variance demonstrated that endotoxin concentration did not change over time, while there was a statistically significant (p < 0.05) decrease in bacterial count over time. In addition, multiple linear regression analysis demonstrated that a unit change in the concentration of endotoxin across time was associated with a significant (p < 0.05) reduction in the bacteriological cell count. This analysis evidenced a significant time effect in the average log bacteriological cell count. Although bacterial growth was not detected in some water samples, endotoxin was present. Measurement of Gram-negative bacterial endotoxins is one of the methods that have been suggested as a rapid way of determining bacteriological water quality.

Microbiological survey of a South African poultry processing plant.

PubMed

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

1995-01-01

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

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.

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.

Microbiological and biochemical effectiveness of an antiseptic gel on the bacterial contamination of the inner space of dental implants: a 3-month human longitudinal study.

PubMed

D'Ercole, S; Tetè, S; Catamo, G; Sammartino, G; Femminella, B; Tripodi, D; Spoto, G; Paolantonio, M

2009-01-01

Microbial penetration inside the implants internal cavity produces a bacterial reservoir that is associated with an area of inflamed connective tissue facing the fixture-abutment junction. The aim of this clinical trial is to evaluate the effectiveness of a 1 percent chlorhexidine gel on the internal bacterial contamination of implants with screw-retained abutments and on the level of AST secreted in peri-implant crevicular fluid. Twenty-five patients (aged 29 to 58 years) each received one implant. Three months after the end of the restorative treatment, and immediately after a clinical and radiographic examination and the abutment removal, microbiological samples were obtained from the internal part of each fixture and biochemical samples were collected by peri-implant sulci. The patients were then divided into two groups: the control (CG; n=10) and test (TG; n=15) groups. The CG had the abutment screwed into place and the crown cemented without any further intervention. In contrast, before the abutment placement and screw tightening, the TG had the internal part of the fixture filled with a 1 percent chlorhexidine gel. Three months later, the same clinical, microbiological and biochemical procedures were repeated in both groups. Total bacterial count, specific pathogens and AST activity were detected. The clinical parameters remained stable throughout the study. From baseline to the 3-month examination, the total bacterial counts underwent a significant reduction only in the TG. In contrast, the AST activity showed a significant increase in the CG. The administration of a 1% chlorhexidine gel appears to be an effective method for the reduction of bacterial colonization of the implant cavity and for safeguarding the health status of peri-implant tissue over a 3-month administration period.

Comparative analyses of viable bacterial counts in foods and seawater under microplate based liquid- and conventional agar plate cultivation: increased culturability of marine bacteria under liquid cultivation.

PubMed

Shigematsu, Toru; Ueno, Shigeaki; Tsuchida, Yasuharu; Hayashi, Mayumi; Okonogi, Hiroko; Masaki, Haruhiko; Fujii, Tomoyuki

2007-12-01

Bacterial counts under liquid cultivation using 96-well microplates were performed. The counts under liquid and under solid cultivation were equivalent in foods, although the counts under liquid cultivation exceeded those under solid cultivation in seawater, suggesting that some bacteria in seawater were viable but did not form detectable colonies. Phylogenetic analysis of bacteria obtained under liquid cultivation was also performed.

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.

Long-term effect of the combined use of powered toothbrush and triclosan dentifrice in periodontal maintenance patients.

PubMed

Bogren, Anna; Teles, Ricardo P; Torresyap, Gay; Haffajee, Anne D; Socransky, Sigmund S; Jönsson, Kerstin; Wennström, Jan L

2008-02-01

To test the hypothesis of a superior clinical and microbiological effect of the combined use of powered toothbrush+triclosan-containing dentifrice compared with manual toothbrush+regular fluoride-containing dentifrice in periodontal maintenance patients. A total of 128 periodontitis subjects involved in recall programmes were randomized to use either powered toothbrush with triclosan-dentifrice (test) or manual toothbrush and standard dentifrice (control). Supportive periodontal treatment was provided at baseline and every 6 months. Plaque, bleeding on probing (BoP), probing pocket depth (PPD) and relative attachment level (RAL) were scored at baseline, 1, 2 and 3 years. Subgingival plaque samples were taken and analysed for their content of 40 bacterial species at each examination interval. All analyses were performed by "intention-to-treat" protocol. Both groups showed significant reduction in BoP, PPD and in mean total counts of the 40 bacterial species between baseline and 3 years, while plaque score and RAL remained almost unchanged. No significant differences between the two prevention programmes were found for any of the clinical outcome variables or in mean counts of the various bacterial species. The study failed to demonstrate superior clinical and microbiological effects of powered toothbrush+triclosan dentifrice compared with manual toothbrush+standard fluoride-dentifrice in periodontitis-susceptible patients on regular maintenance therapy.

Comparing the Effectiveness of Polymer Debriding Devices Using a Porcine Wound Biofilm Model

PubMed Central

Wilkinson, Holly N.; McBain, Andrew J.; Stephenson, Christian; Hardman, Matthew J.

2016-01-01

Objective: Debridement to remove necrotic and/or infected tissue and promote active healing remains a cornerstone of contemporary chronic wound management. While there has been a recent shift toward less invasive polymer-based debriding devices, their efficacy requires rigorous evaluation. Approach: This study was designed to directly compare monofilament debriding devices to traditional gauze using a wounded porcine skin biofilm model with standardized application parameters. Biofilm removal was determined using a surface viability assay, bacterial counts, histological assessment, and scanning electron microscopy (SEM). Results: Quantitative analysis revealed that monofilament debriding devices outperformed the standard gauze, resulting in up to 100-fold greater reduction in bacterial counts. Interestingly, histological and morphological analyses suggested that debridement not only removed bacteria, but also differentially disrupted the bacterially-derived extracellular polymeric substance. Finally, SEM of post-debridement monofilaments showed structural changes in attached bacteria, implying a negative impact on viability. Innovation: This is the first study to combine controlled and defined debridement application with a biologically relevant ex vivo biofilm model to directly compare monofilament debriding devices. Conclusion: These data support the use of monofilament debriding devices for the removal of established wound biofilms and suggest variable efficacy towards biofilms composed of different species of bacteria. PMID:27867752

Feasibility study for epidemic prevention and control in a regional hospital.

PubMed

Chen, Yung-Liang; Yeh, Ming-Yang; Huang, Shau-Yen; Liu, Chi-Ming; Sun, Chi-Chen; Lu, Hsu-Feng; Chiu, Tsan-Hung; Hsia, Te-Chun; Chung, Jing-Gung

2012-03-01

Epidemic prevention policies in hospitals address issues such as, indoor air quality control, cleanliness of medical staff clothing and employee hand-washing procedures. Our hospital employed Bio-Kil to treat air-conditioning filters and nursing staff uniforms. We also assessed the efficacy of different detergents. Using Bio-Kil technology, the mean bacterial count in the air was reduced from 108.8 CFU/h/plate (n=420) to 68.6 CFU/h/plate (n=630). On the lower hems of the Bio-Kil-treated gowns, the mean bacterial count was 1,201 CFU/100 cm(2), markedly lower than the bacterial count of 7,753 CFU/100 cm(2), found on the parts of the gowns not treated with Bio-Kil (p=0.0401). On the cuffs of sleeves treated with Bio-Kil, the mean count was 1,165 CFU/100 cm(2), markedly lower than that of 2,131 CFU/100 cm(2), found on the cuffs not treated with Bio-Kil (p=0.0073). With regard to the mean bacterial eradication rates of antimicrobial solutions, Steridal Solution, 75% alcohol and Bio-Kil (3rd generation) were shown to be the most effective, with rates exceeding 80%. Hibiscrub with paper towels and Fresh Protect Skin were the second most effective. Bio-Kil (1st generation), tap water with paper towels, liquid hand soap with paper towels and ozone water were the least effective. One important observation was that hand-washing without the use of paper towels increased the bacterial count by as much as 84% . Bio-Kil is effective in reducing bacterial counts in the air, on nursing staff uniforms and is an effective detergent.

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

Legionella in industrial cooling towers: monitoring and control strategies.

PubMed

Carducci, A; Verani, M; Battistini, R

2010-01-01

Legionella contamination of industrial cooling towers has been identified as the cause of sporadic cases and outbreaks of legionellosis among people living nearby. To evaluate and control Legionella contamination in industrial cooling tower water, microbiological monitoring was carried out to determine the effectiveness of the following different disinfection treatments: (i) continuous chlorine concentration of 0.01 ppm and monthly chlorine shock dosing (5 ppm) on a single cooling tower; (ii) continuous chlorine concentration of 0.4 ppm and monthly shock of biocide P3 FERROCID 8580 (BKG Water Solution) on seven towers. Legionella spp. and total bacterial count (TBC) were determined 3 days before and after each shock dose. Both strategies demonstrated that when chlorine was maintained at low levels, the Legionella count grew to levels above 10(4) CFU l(-1) while TBC still remained above 10(8 )CFU l(-1). Chlorine shock dosing was able to eliminate bacterial contamination, but only for 10-15 days. Biocide shock dosing was also insufficient to control the problem when the disinfectant concentration was administered at only one point in the plant and at the concentration of 30 ppm. On the other hand, when at a biocide concentration of 30 or 50 ppm was distributed throughout a number of points, depending on the plant hydrodynamics, Legionella counts decreased significantly and often remained below the warning limit. Moreover, the contamination of water entering the plant and the presence of sediment were also important factors for Legionella growth. For effective decontamination of outdoor industrial cooling towers, disinfectants should be distributed in a targeted way, taking into account the possible sources of contamination. The data of the research permitted to modify the procedure of disinfection for better reduce the water and aerosol contamination and consequently the exposure risk.

Comparison of the selection of antimicrobial resistance in fecal Escherichia coli during enrofloxacin administration with a local drug delivery system or with intramuscular injections in a swine model

PubMed Central

Béraud, Romain; Huneault, Louis; Bernier, Dave; Beaudry, Francis; Letellier, Ann; del Castillo, Jérôme R.E.

2008-01-01

This study evaluated, for the first time, the selection of antibiotic resistance in fecal Escherichia coli, a potential reservoir of genes of resistance, during the prolonged exposure to fluoroquinolones after the implantation of a local drug delivery system (LDDS) in a swine model. Fourteen pigs were randomly assigned to group IM (5 mg/kg/day of intramuscular enrofloxacin — EFX) or LD (surgical implantation of EFX-polymethyl-methacrylate perifemoral implants). Blood samples were collected daily for determination of plasma EFX and ciprofloxacin (CFX) concentrations. Fecal samples were collected daily to determine the E. coli counts and the susceptibility patterns of its isolates as evaluated by antibiotic disk diffusion tests. In both groups, EFX administration significantly reduced the bacterial counts after 2 days. During recolonization, the bacterial counts remained lower than baseline in group IM but not significantly, and almost reached pre-treatment levels in group LD. Susceptibility to EFX, CFX, and nalidixic acid of recolonizing E. coli in LD pigs slightly decreased but remained within the limit of “susceptible” isolates. In contrast, quinolone susceptibility of recolonizing E. coli in IM pigs dropped dramatically (P < 0.0001). In addition, intramuscular exposure to fluoroquinolones significantly decreased the susceptibility of E. coli to ampicillin and trimethoprim-sulfamethoxazole (P < 0.05). In conclusion, the use of a dosing regimen that minimized the intestinal output of fluoroquinolones also minimized the selection of resistance to several classes of antibiotics. This could represent another advantage of LDDS usage compared to long-lasting systemic administration of fluoroquinolones. PMID:18783019

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.

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%.

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

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.

[Laboratory diagnosis of bacterial meningitis: usefulness of various tests for the determination of the etiological agent].

PubMed

Carbonnelle, E

2009-01-01

Despite breakthroughs in the diagnosis and treatment of infectious diseases, meningitis still remains an important cause of mortality and morbidity. An accurate and rapid diagnosis of acute bacterial meningitis is essential for a good outcome. The gold-standard test for diagnosis is CSF analysis. Gram staining of CSF reveals bacteria in about 50 to 80 % of cases and cultures are positive in at best 80 % of cases. However, the sensitivity of both tests is less than 50 % in patients who are already on antibiotic treatment. CSF leukocyte count and concentration of protein and glucose lack specificity and sensitivity for the diagnosis of meningitis. Other biological tests are available for the diagnosis. Latex agglutination test were adapted for rapid and direct detection of soluble bacterial antigens in CSF of patients suspected with bacterial meningitis. This test is efficient in detecting antigens of most common central nervous system bateria but lacks sensibility. Furthermore, in the early phases of acute bacterial and viral meningitis, signs and symptoms are often non specific and it is not always possible to make a differential diagnosis. Markers like CRP, procalcitonin, or sTREM-1 may be very useful for the diagnosis and to differentiate between viral and bacterial meningitis. Bacterial meningitis diagnosis and management require various biological tests and a multidisciplinary approach.

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.

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.

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.

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.

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.

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.

A THUMBNAIL HISTORY OF HETEROTROPHIC PLATE COUNT (HPC) METHODOLOGY IN THE UNITED STATES

EPA Science Inventory

Over the past 100 years, the method of determining the number of bacteria in water, foods or other materials has been termed variously as: bacterial plate count, total plate count, total viable plate count, aerobic plate count, standard plate cound and more recently, heterotrophi...

Relationship between Oral Malodor and the Global Composition of Indigenous Bacterial Populations in Saliva ▿

PubMed Central

Takeshita, Toru; Suzuki, Nao; Nakano, Yoshio; Shimazaki, Yoshihiro; Yoneda, Masahiro; Hirofuji, Takao; Yamashita, Yoshihisa

2010-01-01

Oral malodor develops mostly from the metabolic activities of indigenous bacterial populations within the oral cavity, but whether healthy or oral malodor-related patterns of the global bacterial composition exist remains unclear. In this study, the bacterial compositions in the saliva of 240 subjects complaining of oral malodor were divided into groups based on terminal-restriction fragment length polymorphism (T-RFLP) profiles using hierarchical cluster analysis, and the patterns of the microbial community composition of those exhibiting higher and lower malodor were explored. Four types of bacterial community compositions were detected (clusters I, II, III, and IV). Two parameters for measuring oral malodor intensity (the concentration of volatile sulfur compounds in mouth air and the organoleptic score) were noticeably lower in cluster I than in the other clusters. Using multivariate analysis, the differences in the levels of oral malodor were significant after adjustment for potential confounding factors such as total bacterial count, mean periodontal pocket depth, and tongue coating score (P < 0.001). Among the four clusters with different proportions of indigenous members, the T-RFLP profiles of cluster I were implicated as the bacterial populations with higher proportions of Streptococcus, Granulicatella, Rothia, and Treponema species than those of the other clusters. These results clearly correlate the global composition of indigenous bacterial populations with the severity of oral malodor. PMID:20228112

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

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.

Sensitive quantification of Clostridium perfringens in human feces by quantitative real-time PCR targeting alpha-toxin and enterotoxin genes.

PubMed

Nagpal, Ravinder; Ogata, Kiyohito; Tsuji, Hirokazu; Matsuda, Kazunori; Takahashi, Takuya; Nomoto, Koji; Suzuki, Yoshio; Kawashima, Kazunari; Nagata, Satoru; Yamashiro, Yuichiro

2015-10-19

Clostridium perfringens is a widespread pathogen, but the precise quantification of this subdominant gut microbe remains difficult due to its low fecal count (particularly in asymptomatic subjects) and also due to the presence of abundant polymerase-inhibitory substances in human feces. Also, information on the intestinal carriage of toxigenic C. perfringens strains in healthy subjects is sparse. Therefore, we developed a sensitive quantitative real-time PCR assays for quantification of C. perfringens in human feces by targeting its α-toxin and enterotoxin genes. To validate the assays, we finally observed the occurrence of α-toxigenic and enterotoxigenic C. perfringens in the fecal microbiota of healthy Japanese infants and young adults. The plc-specific qPCR assay was newly validated, while primers for 16S rRNA and cpe genes were retrieved from literature. The assays were validated for specificity and sensitivity in pre-inoculated fecal samples, and were finally applied to quantify C. perfringens in stool samples from apparently healthy infants (n 124) and young adults (n 221). The qPCR assays were highly specific and sensitive, with a minimum detection limit of 10(3) bacterial cells/g feces. Alpha-toxigenic C. perfringens was detected in 36% infants and 33% adults, with counts ranging widely (10(3)-10(7) bacterial cells/g). Intriguingly, the mean count of α-toxigenic C. perfringens was significantly higher in infants (6.0±1.5 log10 bacterial cells/g), as compared to that in adults (4.8±1.2). Moreover, the prevalence of enterotoxigenic C. perfringens was also found to be significantly higher in infants, as compared to that in adults. The mean enterotoxigenic C. perfringens count was 5.9±1.9 and 4.8±0.8 log10 bacterial cells/g in infants and adults, respectively. These data indicate that some healthy infants and young adults carry α-toxigenic and enterotoxigenic C. perfringens at significant levels, and may be predisposed to related diseases. Thus, high fecal carriage of toxigenic C. perfringens in healthy children warrants further investigation on its potential sources and clinical significance in these subjects. In summary, we present a novel qPCR assay for sensitive and accurate quantification of α-toxigenic and enterotoxigenic C. perfringens in human feces, which should facilitate prospective studies of the gut microbiota.

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.

Rapid staining and enumeration of small numbers of total bacteria in water by solid-phase laser cytometry

NASA Technical Reports Server (NTRS)

Broadaway, Susan C.; Barton, Stephanie A.; Pyle, Barry H.

2003-01-01

The nucleic acid stain SYBR Green I was evaluated for use with solid-phase laser cytometry to obtain total bacterial cell counts from several water sources with small bacterial numbers. Results were obtained within 30 min and exceeded or equaled counts on R2A agar plates incubated for 14 days at room temperature.

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.

What's growing on your stethoscope? (And what you can do about it).

PubMed

Schroeder, Ariel; Schroeder, Maryellen A; D'Amico, Frank

2009-08-01

Studies have shown that rubbing alcohol pads on stethoscope diaphragms can reduce bacterial colonization, but alcohol pads are used infrequently used and not always available. We conducted a prospective, single-blinded study to investigate whether simultaneously scrubbing hands and stethoscope head with alcohol-based hand foam would significantly reduce bacterial counts on the stethoscope. Using their own stethoscope, participants imprinted the stethoscope head onto a chocolate agar plate, then used alcohol-based hand foam to cleanse their hands while simultaneously rubbing the stethoscope head. Once the stethoscope heads were dry, the participants imprinted their stethoscope heads onto a second plate. After 48 hours' incubation, we determined the bacterial counts for the prewash and post-wash plates, and compared the 2. We analyzed a total of 184 cultures (from 92 stethoscopes). Both the mean (28 prewash vs 3 post-wash, P=.001) and median (11 prewash vs 1 post-wash, P=.001) colony counts were significantly greater before being cleansed. Three methicillin-resistant Staphylococcus aureus (MRSA) colonies were identified in the prewash period; all were destroyed by the foam. The estimated number of hand washes needed to prevent 1 MRSA colony is 31 (95% confidence interval [CI], 18-89). Simultaneously using hand foam to clean hands and stethoscope heads reduces bacterial counts on stethoscopes. Further research is needed to determine whether this intervention can reduce morbidity and mortality associated with bacterial infection.

Sonication reduces the attachment of Salmonella Typhimurium ATCC 14028 cells to bacterial cellulose-based plant cell wall models and cut plant material.

PubMed

Tan, Michelle S F; Rahman, Sadequr; Dykes, Gary A

2017-04-01

This study investigated the removal of bacterial surface structures, particularly flagella, using sonication, and examined its effect on the attachment of Salmonella Typhimurium ATCC 14028 cells to plant cell walls. S. Typhimurium ATCC 14028 cells were subjected to sonication at 20 kHz to remove surface structures without affecting cell viability. Effective removal of flagella was determined by staining flagella of sonicated cells with Ryu's stain and enumerating the flagella remaining by direct microscopic counting. The attachment of sonicated S. Typhimurium cells to bacterial cellulose-based plant cell wall models and cut plant material (potato, apple, lettuce) was then evaluated. Varying concentrations of pectin and/or xyloglucan were used to produce a range of bacterial cellulose-based plant cell wall models. As compared to the non-sonicated controls, sonicated S. Typhimurium cells attached in significantly lower numbers (between 0.5 and 1.0 log CFU/cm 2 ) to all surfaces except to the bacterial cellulose-only composite without pectin and xyloglucan. Since attachment of S. Typhimurium to the bacterial cellulose-only composite was not affected by sonication, this suggests that bacterial surface structures, particularly flagella, could have specific interactions with pectin and xyloglucan. This study indicates that sonication may have potential applications for reducing Salmonella attachment during the processing of fresh produce. Copyright © 2016 Elsevier Ltd. All rights reserved.

Microbial dynamics during production of lesser mealworms (Alphitobius diaperinus) for human consumption at industrial scale.

PubMed

Wynants, E; Crauwels, S; Verreth, C; Gianotten, N; Lievens, B; Claes, J; Van Campenhout, L

2018-04-01

In this study, the microbial dynamics during an industrial production cyle of lesser mealworms (Alphitobius diaperinus), sold for human consumption, were characterised. The microbial numbers as well as the microbial diversity were generally higher for the substrate, existing of remaining feed, faeces and exuviae, than for the larvae. Most of the species-level operational taxonomic units, identified using Illumina MiSeq sequencing, that were present in the feed were also detected in the larvae and vice versa. However, bacterial diversity decreased in the larvae during rearing. These results suggested that the feed is an important determinant of the insect bacterial community, but that some bacterial species show a competitive advantage inside the insect gut and become dominant. A blanching treatment of the larvae after harvest reduced most microbial counts, but the number of aerobic endospores remained at 4.0 log cfu/g. Whereas food pathogens Salmonella spp., Listeria monocytogenes, Bacillus cereus or coagulase-positive staphylococci were not detected in our study, fungal isolates corresponding to the genera Aspergillus and Fusarium were recovered. Therefore, it cannot be excluded that mycotoxins were present. The results of this study contribute to a better understanding of the microbial dynamics and food safety aspects during the production of edible insects. Copyright © 2017 Elsevier Ltd. All rights reserved.

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.

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.

[Efficacy of five disinfectants to reduce bacterial load in the household].

PubMed

Stambullian, Julián; Rossotti, Daniel; Fridman, Diego; Luchetti, Pablo; Cheade, Yamila; Stamboulian, Daniel

2011-01-01

The proper use of products containing sodium hypochlorite,ammonium salts and triclosan has proved to be effective in the elimination of infectious agents in the household environment. Our objective was to evaluate the immediate, one-week and one-month efficacy of controlled use of five products containing these components, compared to other commonly used products. Within a six month period, thirty two middle-class homes from Buenos Aires City and suburbs were included in this open-label, randomized, parallel-group intervention study. Sixteen homes were randomized to use products containing sodium hypochlorite, ammonia and triclosan in the kitchen and bathroom during one month. The remaining maintained usual practices for domestic cleaning. Bacterial counts and identification were performed from samples taken from each study site. Baseline samples (no group discrimination) contained a mean bacterial count in kitchen of 66.0 CFU/cm2, and in bathroom 40.1 CFU/cm2. Samples taken immediately after-cleaning (no group discrimination) contained: kitchen 0.8 CFU/cm2; bathroom < 1 CFU/cm2. After one week (intervention group vs. control group) contained: kitchen 18.0 vs. 32.5 CFU/cm2; bathroom 12.7 vs. 7.7 CFU/cm2. After one month (intervention group vs. control group): kitchen 60.1 vs. 62.1 CFU/cm2; bathroom 37.0 vs. 42.0 CFU/cm2. A remarkable decrease of bacterial load was observed in both groups, which suggests that not only product quality but also education for suitable use plays a key role in successful house disinfection. This approach could be an important tool for improving prevention of foodborne infections since fecal coliforms widely predominated in all analyzed samples.

Variation in antibiotic-induced microbial recolonization impacts on the host metabolic phenotypes of rats.

PubMed

Swann, Jonathan R; Tuohy, Kieran M; Lindfors, Peter; Brown, Duncan T; Gibson, Glenn R; Wilson, Ian D; Sidaway, James; Nicholson, Jeremy K; Holmes, Elaine

2011-08-05

The interaction between the gut microbiota and their mammalian host is known to have far-reaching consequences with respect to metabolism and health. We investigated the effects of eight days of oral antibiotic exposure (penicillin and streptomycin sulfate) on gut microbial composition and host metabolic phenotype in male Han-Wistar rats (n = 6) compared to matched controls. Early recolonization was assessed in a third group exposed to antibiotics for four days followed by four days recovery (n = 6). Fluorescence in situ hybridization analysis of the intestinal contents collected at eight days showed a significant reduction in all bacterial groups measured (control, 10(10.7) cells/g feces; antibiotic-treated, 10(8.4)). Bacterial suppression reduced the excretion of mammalian-microbial urinary cometabolites including hippurate, phenylpropionic acid, phenylacetylglycine and indoxyl-sulfate whereas taurine, glycine, citrate, 2-oxoglutarate, and fumarate excretion was elevated. While total bacterial counts remained notably lower in the recolonized animals (10(9.1) cells/g faeces) compared to the controls, two cage-dependent subgroups emerged with Lactobacillus/Enterococcus probe counts dominant in one subgroup. This dichotomous profile manifested in the metabolic phenotypes with subgroup differences in tricarboxylic acid cycle metabolites and indoxyl-sulfate excretion. Fecal short chain fatty acids were diminished in all treated animals. Antibiotic treatment induced a profound effect on the microbiome structure, which was reflected in the metabotype. Moreover, the recolonization process was sensitive to the microenvironment, which may impact on understanding downstream consequences of antibiotic consumption in human populations.

Microbiological hazard identification and exposure assessment of street food vending in Johannesburg, South Africa.

PubMed

Mosupye, F M; von Holy, A

2000-11-01

One hundred and thirty-two samples of beef, chicken, salad and gravy were collected from two street vendors over eleven replicate surveys to assess microbiological safety and quality. For each food type samples were collected during preparation and holding. Dish water was also collected and food preparation surfaces swabbed during preparation and display. Standard methods were used to determine aerobic plate counts, Enterobacteriaceae counts, coliform counts and spore counts. Six hundred and seventy-five predominant colonies were isolated from aerobic plate counts of all samples and characterised. The incidence of selected foodborne bacterial pathogens and non-pathogenic E. coli 1 was also determined. In most cases mean bacterial counts of the raw materials were significantly higher (P < 0.05) than those of corresponding cooked foods. No significant differences (P > 0.05) in all count types were observed between food samples collected during cooking and those collected during holding. In addition, no significant differences (P > 0.05) in all count types were observed between prepared salads and their raw materials. Mean bacterial counts of water and swab samples collected from vendor 1 were lower than those of water and swab samples collected from vendor 2.The predominant populations isolated from the aerobic plate counts were Bacillus spp., Staphylococcus spp., Enterobacteriaceae and Alcaligenes spp. Bacillus cereus was detected in 17%, Clostridium perfringens in 1%, Staphylococcus aureus in 3% and Vibrio metchnikovii in 2% of the food samples. Campylobacter jejuni, Listeria monocytogenes, and Escherichia coli O157:H7 were not detected. Non-pathogenic E. coli 1 was detected in 13% of food samples, in 86 and 36% of dish water samples collected from vendors 1 and 2, respectively, and in 36% of surface swab samples from vendor 2.

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.

Can Particulate Air Sampling Predict Microbial Load in Operating Theatres for Arthroplasty?

PubMed Central

Cristina, Maria Luisa; Spagnolo, Anna Maria; Sartini, Marina; Panatto, Donatella; Gasparini, Roberto; Orlando, Paolo; Ottria, Gianluca; Perdelli, Fernanda

2012-01-01

Several studies have proposed that the microbiological quality of the air in operating theatres be indirectly evaluated by means of particle counting, a technique derived from industrial clean-room technology standards, using airborne particle concentration as an index of microbial contamination. However, the relationship between particle counting and microbiological sampling has rarely been evaluated and demonstrated in operating theatres. The aim of the present study was to determine whether particle counting could predict microbiological contamination of the air in an operating theatre during 95 surgical arthroplasty procedures. This investigation was carried out over a period of three months in 2010 in an orthopedic operating theatre devoted exclusively to prosthetic surgery. During each procedure, the bacterial contamination of the air was determined by means of active sampling; at the same time, airborne particulate contamination was assessed throughout the entire procedure. On considering the total number of surgical operations, the mean value of the total bacterial load in the center of the operating theatre proved to be 35 CFU/m3; the mean particle count was 4,194,569 no./m3 for particles of diameter ≥0.5 µm and 13,519 no./m3 for particles of diameter ≥5 µm. No significant differences emerged between the median values of the airborne microbial load recorded during the two types of procedure monitored. Particulates with a diameter of ≥0.5 µm were detected in statistically higher concentrations (p<0.001) during knee-replacement procedures. By contrast, particulates with a diameter of ≥5 µm displayed a statistically higher concentration during hip-replacement procedures (p<0.05). The results did not reveal any statistically significant correlation between microbial loads and particle counts for either of the particle diameters considered (≥0.5 µm and ≥5 µm). Consequently, microbiological monitoring remains the most suitable method of evaluating the quality of air in operating theatres. PMID:23285189

Influence of raw milk quality on fluid milk shelf life.

PubMed

Barbano, D M; Ma, Y; Santos, M V

2006-03-01

Pasteurized fluid milk shelf life is influenced by raw milk quality. The microbial count and somatic cell count (SCC) determine the load of heat-resistant enzymes in milk. Generally, high levels of psychrotrophic bacteria in raw milk are required to contribute sufficient quantities of heat-stable proteases and lipases to cause breakdown of protein and fat after pasteurization. Sanitation, refrigeration, and the addition of CO2 to milk are used to control both total and psychrotrophic bacteria count. It is not uncommon for total bacterial counts of raw milk to be < 10,000 cfu/mL. In the past, fluid milk processors have not focused much attention on milk SCC. Increased SCC is correlated with increased amounts of heat-stable protease (plasmin) and lipase (lipoprotein lipase) in milk. When starting with raw milk that has a low bacterial count, and in the absence of microbial growth in pasteurized milk, enzymes associated with high SCC will cause protein and fat degradation during refrigerated storage, and produce off-flavors. As the ability to kill, remove, or control microbial growth in pasteurized refrigerated milk continues to improve, the original milk SCC will be the factor limiting the time of refrigerated storage before development of an off-flavor in milk. Most healthy cows in a dairy herd have a milk SCC < 50,000 cell/mL. Bulk tank SCC > 200,000 cell/mL are usually due to the contribution of high SCC milk from a small number of cows in the herd. Technology to identify these cows and keep their milk out of the bulk tank could substantially increase the value of the remaining milk for use in fluid milk processing. To achieve a 60- to 90-d shelf life of refrigerated fluid milk, fluid processors and dairy farmers need to work together to structure economic incentives that allow farmers to produce milk with the SCC needed for extended refrigerated shelf life.

Can particulate air sampling predict microbial load in operating theatres for arthroplasty?

PubMed

Cristina, Maria Luisa; Spagnolo, Anna Maria; Sartini, Marina; Panatto, Donatella; Gasparini, Roberto; Orlando, Paolo; Ottria, Gianluca; Perdelli, Fernanda

2012-01-01

Several studies have proposed that the microbiological quality of the air in operating theatres be indirectly evaluated by means of particle counting, a technique derived from industrial clean-room technology standards, using airborne particle concentration as an index of microbial contamination. However, the relationship between particle counting and microbiological sampling has rarely been evaluated and demonstrated in operating theatres. The aim of the present study was to determine whether particle counting could predict microbiological contamination of the air in an operating theatre during 95 surgical arthroplasty procedures. This investigation was carried out over a period of three months in 2010 in an orthopedic operating theatre devoted exclusively to prosthetic surgery. During each procedure, the bacterial contamination of the air was determined by means of active sampling; at the same time, airborne particulate contamination was assessed throughout the entire procedure. On considering the total number of surgical operations, the mean value of the total bacterial load in the center of the operating theatre proved to be 35 CFU/m(3); the mean particle count was 4,194,569 no./m(3) for particles of diameter ≥0.5 µm and 13,519 no./m(3) for particles of diameter ≥5 µm. No significant differences emerged between the median values of the airborne microbial load recorded during the two types of procedure monitored. Particulates with a diameter of ≥0.5 µm were detected in statistically higher concentrations (p<0.001) during knee-replacement procedures. By contrast, particulates with a diameter of ≥5 µm displayed a statistically higher concentration during hip-replacement procedures (p<0.05). The results did not reveal any statistically significant correlation between microbial loads and particle counts for either of the particle diameters considered (≥0.5 µm and ≥5 µm). Consequently, microbiological monitoring remains the most suitable method of evaluating the quality of air in operating theatres.

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.

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.

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.

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.

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.

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.

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.

Viral-bacterial coinfection affects the presentation and alters the prognosis of severe community-acquired pneumonia.

PubMed

Voiriot, Guillaume; Visseaux, Benoit; Cohen, Johana; Nguyen, Liem Binh Luong; Neuville, Mathilde; Morbieu, Caroline; Burdet, Charles; Radjou, Aguila; Lescure, François-Xavier; Smonig, Roland; Armand-Lefèvre, Laurence; Mourvillier, Bruno; Yazdanpanah, Yazdan; Soubirou, Jean-Francois; Ruckly, Stephane; Houhou-Fidouh, Nadhira; Timsit, Jean-François

2016-10-25

Multiplex polymerase chain reaction (mPCR) enables recovery of viruses from airways of patients with community-acquired pneumonia (CAP), although their clinical impact remains uncertain. Among consecutive adult patients who had undergone a mPCR within 72 hours following their admission to one intensive care unit (ICU), we retrospectively included those with a final diagnosis of CAP. Four etiology groups were clustered: bacterial, viral, mixed (viral-bacterial) and no etiology. A composite criterion of complicated course (hospital death or mechanical ventilation > 7 days) was used. A subgroup analysis compared patients with bacterial and viral-bacterial CAP matched on the bacterial pathogens. Among 174 patients (132 men [76 %], age 63 [53-75] years, SAPSII 38 [27;55], median PSI score 106 [78;130]), bacterial, viral, mixed and no etiology groups gathered 46 (26 %), 53 (31 %), 45 (26 %) and 30 (17 %) patients, respectively. Virus-infected patients displayed a high creatine kinase serum level, a low platelet count, and a trend toward more frequent alveolar-interstitial infiltrates. A complicated course was more frequent in the mixed group (31/45, 69 %), as compared to bacterial (18/46, 39 %), viral (15/53, 28 %) and no etiology (12/30, 40 %) groups (p < 0.01). In multivariate analysis, the mixed (viral-bacterial) infection was independently associated with complicated course (reference: bacterial pneumonia; OR, 3.58; CI 95 %, 1.16-11; p = 0.03). The subgroup analysis of bacteria-matched patients confirmed these findings. Viral-bacterial coinfection during severe CAP in adults is associated with an impaired presentation and a complicated course.

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.

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.

Bacterial screening by flow cytometry offers potential for extension of platelet storage: results of 14 months of active surveillance.

PubMed

Vollmer, T; Engemann, J; Kleesiek, K; Dreier, J

2011-06-01

Bacterial contamination is currently the major infectious hazard of platelet transfusion in developed countries. It has been demonstrated that a significant transfusion risk remains, in particular with older platelet concentrates (PCs). In 2009, the shelf life of PCs was therefore reduced in Germany to 4 days after the day of production according to Vote 38. The aim of the present study was the application and implementation of a recently developed flow cytometry-based rapid screening method (BactiFlow) for bacterial contamination at the end of PC shelf life as a routine in-process control. A total of 472 apheresis-derived PCs were tested using the BactiFlow flow cytometric assay to detect and count bacteria based on esterase activity in viable bacterial cells, while the BacT/Alert automated culture system served as the reference method. The automation potential of the flow cytometric assay was analysed by applying the semi-automated BactiFlow ALS system. An algorithm was developed for use in routine blood bank operations to extend the storage period of PCs. Two of the 472 apheresis PCs tested were positive in culture and identified as Propionibacterium species. One PC was positive for Staphylococcus aureus by both methods. All remaining specimens were tested negative by both methods. Our study demonstrates that routine bacterial testing of PCs was successfully implemented and the established algorithm proved efficient. The BactiFlow flow cytometric assay is the first rapid screening method which is suitable for a routine application combined with a high sensitivity. © 2011 The Authors. Transfusion Medicine © 2011 British Blood Transfusion Society.

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.

Validation of an automated colony counting system for group A Streptococcus.

PubMed

Frost, H R; Tsoi, S K; Baker, C A; Laho, D; Sanderson-Smith, M L; Steer, A C; Smeesters, P R

2016-02-08

The practice of counting bacterial colony forming units on agar plates has long been used as a method to estimate the concentration of live bacteria in culture. However, due to the laborious and potentially error prone nature of this measurement technique, an alternative method is desirable. Recent technologic advancements have facilitated the development of automated colony counting systems, which reduce errors introduced during the manual counting process and recording of information. An additional benefit is the significant reduction in time taken to analyse colony counting data. Whilst automated counting procedures have been validated for a number of microorganisms, the process has not been successful for all bacteria due to the requirement for a relatively high contrast between bacterial colonies and growth medium. The purpose of this study was to validate an automated counting system for use with group A Streptococcus (GAS). Twenty-one different GAS strains, representative of major emm-types, were selected for assessment. In order to introduce the required contrast for automated counting, 2,3,5-triphenyl-2H-tetrazolium chloride (TTC) dye was added to Todd-Hewitt broth with yeast extract (THY) agar. Growth on THY agar with TTC was compared with growth on blood agar and THY agar to ensure the dye was not detrimental to bacterial growth. Automated colony counts using a ProtoCOL 3 instrument were compared with manual counting to confirm accuracy over the stages of the growth cycle (latent, mid-log and stationary phases) and in a number of different assays. The average percentage differences between plating and counting methods were analysed using the Bland-Altman method. A percentage difference of ±10 % was determined as the cut-off for a critical difference between plating and counting methods. All strains measured had an average difference of less than 10 % when plated on THY agar with TTC. This consistency was also observed over all phases of the growth cycle and when plated in blood following bactericidal assays. Agreement between these methods suggest the use of an automated colony counting technique for GAS will significantly reduce time spent counting bacteria to enable a more efficient and accurate measurement of bacteria concentration in culture.

Efficacy of chlorinated and ozonated water in reducing Salmonella typhimurium attached to tomato surfaces.

PubMed

Chaidez, Cristobal; Lopez, Javier; Vidales, Juan; Campo, Nohelia Castro-Del

2007-08-01

The purpose of this study was to compare chlorinated and ozonated water in reducing Salmonella typhimurium inoculated onto fresh ripe tomatoes. Surface-inoculated tomatoes were immersed/sprayed with chlorinated (200 mg l(-1)) and ozonated water (1 and 2 mg l(-1)) under 2 and 100 nefelometric turbidity units (NTU). Contact times were 120 and 30 s for immersing and spraying applications, respectively. Immersing in chlorinated water and low turbidity resulted in the most effective application with 3.61 log(10) bacterial reduction, while 1 and 2 mg l(-1) of ozone reduced 2.32 and 2.53 log(10), respectively. High turbidity and chlorine reduced the bacterial counts by 3.39 log(10), while 1 and 2 mg l(-1) of ozonated water and low turbidity reduced the bacteria by 1.48 and 1.92 log(10), respectively. Spraying chlorinated water reduced bacteria by 3 log(10), and ozonated water at 1 and 2 mg l(-1) reduced counts by 1.84 and 2.40 log(10), respectively. No statistical differences were found between chlorine and ozone (2 mg l(-1)) during spraying applications (p < 0.05). The use of ozonated water both in immersing and spraying applications is suggested when water turbidity remains low.

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.

Diagnosis of Streptococcus pneumoniae and Haemophilus influenzae type b meningitis by identifying DNA from cerebrospinal fluid-impregnated filter paper strips.

PubMed

Peltola, Heikki; Roine, Irmeli; Leinonen, Maija; Kuisma, Leena; Mata, Antonio González; Arbo, Antonio; Goyo, José; Saukkoriipi, Annika

2010-02-01

Bacterial meningitis remains often etiologically unconfirmed, especially in resource-poor settings. We tested the potential of real-time polymerase chain reaction to identify Streptococcus pneumoniae (Pnc) and Haemophilus influenzae type b (Hib) from cerebrospinal fluid impregnated on filter paper strips. Pnc and Hib genome equivalents were blindly quantified by polymerase chain reaction from 129 liquid cerebrospinal fluid (CSF) samples-the standard-and strips stored at room temperature for months. Genome counts were compared by simple regression. The strips showed a sensitivity and specificity of 92% and 99% for Pnc, and of 70% and 100% for Hib, respectively. The positive and negative predictive values were 94% and 97% for Pnc, and 100% and 89% for Hib, respectively. For Pnc, the positive and negative likelihood ratio was 92 and 0.08, and the overall accuracy 98%, whereas for Hib they were 70 and 0.30, and 91%, respectively. Genome counting showed good correlation between the filter paper and liquid CSF samples, r(2) being 0.87 for Pnc and 0.68 for Hib (P < 0.0001 for both). Although not replacing bacterial culture, filter paper strips offer an easy way to collect and store CSF samples for later bacteriology. They can also be transported in standard envelops by regular mail.

Fermentation characteristics and angiotensin I-converting enzyme-inhibitory activity of Lactobacillus helveticus isolate H9 in cow milk, soy milk, and mare milk.

PubMed

Wang, Jicheng; Li, Changkun; Xue, Jiangang; Yang, Jie; Zhang, Qing; Zhang, Heping; Chen, Yongfu

2015-06-01

Lactobacillus helveticus isolate H9 demonstrated high angiotensin I-converting enzyme (ACE)-inhibitory activity in previous research. Here, we evaluated the fermentation characteristics (pH, titratable acidity, free amino nitrogen, and viable bacterial counts), ACE-inhibitory activity, and contents of Val-Pro-Pro (VPP) and Ile-Pro-Pro (IPP) peptides of stored yogurt (4°C for 28 d) fermented by L. helveticus isolate H9 (initially inoculated at 4 concentrations), from cow, mare, and soy milks. During storage, the pH and titratable acidity remained stable in yogurts produced from all milk types and all inoculation concentrations. The viable bacterial counts in all stored yogurts ranged between 10(6.72) and 10(8.59) cfu/g. The highest ACE-inhibitory activity (70.9-74.5%) was achieved at inoculation concentrations of 5×10(6) cfu/mL. The ACE-inhibitory tripeptides VPP and IPP as determined by ultra-performance liquid chromatography-tandem mass spectrometry were not produced in yogurt made from soy milk or mare milk. These evaluations indicate that L. helveticus H9 has good probiotic properties and would be a promising candidate for production of fermented food with probiotic properties. Copyright © 2015 American Dairy Science Association. Published by Elsevier Inc. All rights reserved.

Performance and Longevity of a Novel Intraosseous Device in a Goat (Capra hircus) Model

PubMed Central

Jackson, Erin E; Ashley, T Clay; Snowden, Karen F; Gresham, Vincent C; Budke, Christine M; Eichelberger, Bunita M; Taylor, Destiny A

2011-01-01

We performed 2 studies to assess the function and longevity of a novel intraosseous catheter device. For study 1, 9 goats were assigned to 3 groups (intraosseous catheter in the proximal humerus, intraosseous catheter in the proximal tibia, or standard jugular catheter). Devices in the tibia remained in place for less time than did those in the humerus, and no goats exhibited radiographic evidence of resulting damage or structural change in surrounding bone. Positive bacterial cultures were found in all 9 goats at various time points. In study 2, 18 goats were assigned to 2 groups (intraosseous catheter in the wing of the ilium or proximal humerus). Samples for serial aerobic and anaerobic blood cultures and CBC were collected while devices remained in use. Clinical monitoring and removal criteria were identical those for study 1. Catheters in the ilium remained in place for less than 24 h on average, and those in the humerus remained in place for an average of 2.5 d. Several goats with proximal humeral catheters demonstrated moderate lameness after removal, and radiographic evidence of periosteal bone growth was noted in another goat. Bloodwork indicated mild elevations of WBC counts from baseline in some cases. Bacterial growth was found in samples from 4 of 18 goats at various time points. Our study indicated that intraosseous catheters may remain safely in place for more than 24 h, but animals should be monitored closely for negative side effects for several days after removal. PMID:21640033

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.

Back to the Basics: Community-Acquired Pneumonia in Children.

PubMed

Boyd, Kathleen

2017-07-01

Community-acquired pneumonia (CAP) is a common childhood infection and often a reason for inpatient admission, especially when a child is hypoxic or in respiratory distress. Despite advances in technology and diagnostics, it remains difficult to accurately differentiate bacterial CAP from a viral process. Most of the laboratory tests routinely done in inpatient medicine, such as complete blood counts and acute phase reactants, do little to differentiate a viral pneumonia from a bacterial pneumonia. Clinicians must rely heavily on the clinical presentation and decide whether to treat empirically with antibiotics. Guidelines published by the Infectious Disease Society of America in 2011 have helped clinicians standardize the diagnosis and treatment of CAP. The guidelines recommend relatively narrow-spectrum antibiotics, such as ampicillin or penicillin, as empiric coverage for the fully immunized child older than age 3 months who requires hospitalization for CAP. [Pediatr Ann. 2017;46(7):e257-e261.]. Copyright 2017, SLACK Incorporated.

Kinetics of bacterial fluorescence staining with 3,3'-diethylthiacyanine.

PubMed

Thomas, Marlon S; Nuñez, Vicente; Upadhyayula, Srigokul; Zielins, Elizabeth R; Bao, Duoduo; Vasquez, Jacob M; Bahmani, Baharak; Vullev, Valentine I

2010-06-15

For more than a century, colorimetric and fluorescence staining have been the foundation of a broad range of key bioanalytical techniques. The dynamics of such staining processes, however, still remains largely unexplored. We investigated the kinetics of fluorescence staining of two gram-negative and two gram-positive species with 3,3'-diethylthiacyanine (THIA) iodide. An increase in the THIA fluorescence quantum yield, induced by the bacterial dye uptake, was the principal reason for the observed emission enhancement. The fluorescence quantum yield of THIA depended on the media viscosity and not on the media polarity, which suggested that the microenvironment of the dye molecules taken up by the cells was restrictive. The kinetics of fluorescence staining did not manifest a statistically significant dependence neither on the dye concentration, nor on the cell count. In the presence of surfactant additives, however, the fluorescence-enhancement kinetic patterns manifested species specificity with statistically significant discernibility.

Photoperiod but not food restriction modulates innate immunity in an opportunistic breeder, Loxia curvirostra.

PubMed

Schultz, Elizabeth M; Hahn, Thomas P; Klasing, Kirk C

2017-02-15

An organism's investment in immune function often varies seasonally but understanding of how fluctuations in environmental conditions directly modulate investment remains limited. This experiment investigated how changes in photoperiod and food availability affect investment in constitutive innate immunity and the acute phase response induced by lipopolysaccharide (LPS) injections in captive red crossbills ( Loxia curvirostra ). Crossbills are reproductively flexible songbirds that specialize on an unpredictably available food resource and display temporal variation in immunity in the wild. Birds were separated into four treatments and exposed to long or short day lengths for 6 weeks before continuing on an ad libitum diet or experiencing a 20% food reduction for 10 days. Birds were un-injected or injected with LPS both before and after diet change. Innate immunity was quantified throughout the experiment to assess effects of photoperiod, food availability and their interactions on hemolysis-hemagglutination, haptoglobin, bacterial killing ability and leukocyte counts. Overall, increasing day length significantly increased both bacterial killing ability and leukocyte counts. Surprisingly, food restriction had little effect on the immune parameters, potentially owing to the 'low-cost' environment of captivity and suggesting that investment in innate immunity is prioritized and maintained whenever possible. LPS injections induced stereotypical sickness behaviors and increased bacterial killing ability in short day birds and complement activity (hemolysis) both before and after food restriction. These results demonstrate robust seasonal modulation of immune investment and an ability to maintain innate immunity in the face of limited resources in these temporally flexible songbirds. © 2017. Published by The Company of Biologists Ltd.

Diagnosis of nosocomial pneumonia in intubated, intensive care unit patients.

PubMed

Salata, R A; Lederman, M M; Shlaes, D M; Jacobs, M R; Eckstein, E; Tweardy, D; Toossi, Z; Chmielewski, R; Marino, J; King, C H

1987-02-01

The clinical distinction between bacterial colonization of the tracheobronchial tree and nosocomial pneumonia is difficult, especially in intubated patients. We studied 51 intubated, intensive care unit patients prospectively by serial examinations of tracheal aspirates for elastin fibers, graded Gram's stains, and quantitative bacterial cultures in conjunction with clinical and radiologic observations in an attempt to develop criteria for the early detection of pulmonary infection. Patients with infection had new or progressive pulmonary infiltrates plus 1 of the following: positive blood culture results, radiographic evidence of cavitation, or histologic evidence of pneumonia, or 2 or more of the following: new fever, new leukocytosis, or grossly purulent tracheal aspirates. Twenty-one patients developed infection, 22 remained colonized, and 8 had an uncertain status. Infiltrates developed in 34 patients (21 infected, 8 colonized, 5 uncertain status). Gram-negative bacilli were most commonly isolated and were more frequent in infected patients (81 versus 47%, p less than 0.05); Pseudomonas aeruginosa and Serratia marcescens were most often associated with infection. No differences were observed between infected and colonized patients in demographic features, smoking history, underlying disease, previous antibiotic therapy, days in hospital before intubation, preexisting pneumonia upon intubation, or highest temperature or leukocyte count during course. By univariate analysis, infected patients had a longer duration of intubation (p less than 0.05), higher Gram's stain grading for neutrophils (p less than 0.05) or bacteria (p less than 0.005), higher bacterial colony counts (p less than 0.05), and more frequent detection of elastin fibers in tracheal aspirates (p less than 0.02).(ABSTRACT TRUNCATED AT 250 WORDS)

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.

[Effects of grape seed addition in swine manure-wheat straw composting on the compost microbial community and carbon and nitrogen contents].

PubMed

Huang, Yi-Mei; Liu, Xue-Ling; Jiang, Ji-Shao; Huang, Hua; Liu, Dong

2012-08-01

Taking substrates swine manure and wheat straw (fresh mass ratio 10.5:1) as the control (PMW), a composting experiment was conducted in a self-made aerated static composting bin to study the effects of adding 8% grape seed (treatment PMW + G) on the succession of microbial community and the transformation of carbon and nitrogen in the substrates during the composting. Seven samples were collected from each treatment, according to the temperature of the compost during the 30 d composting period. The microbial population and physiological groups were determined, and the NH4(+)-N, NO3(-)-N, organic N, and organic C concentrations in the compost were measured. Grape seed addition induced a slight increase of bacterial count and a significant increase of actinomycetes count, but decreased the fungal count significantly. Grape seed addition also decreased the ratio of bacteria to actinomycetes and the counts of ammonifiers and denitrifiers, but increased the counts of nitrifiers, N-fixing bacteria, and cellulose-decomposing microorganisms. The contents of NH4(+)-N and organic C decreased, while that of NO3(-)-N increased obviously. The NO3(-)-N content in the compost was positively correlated with the actinomycetes count. During composting, the compost temperature in treatment PMW + G increased more rapidly, and remained steady in thermophilic phase, while the water content changed little, which provided a stable and higher population of actinomycetes and nitrifiers in thermophilic phase, being beneficial to the increase of compost nitrate N.

Comparison of plate counts, Petrifilm, dipslides, and adenosine triphosphate bioluminescence for monitoring bacteria in cooling-tower waters.

PubMed

Mueller, Sherry A; Anderson, James E; Kim, Byung R; Ball, James C

2009-04-01

Effective bacterial control in cooling-tower systems requires accurate and timely methods to count bacteria. Plate-count methods are difficult to implement on-site, because they are time- and labor-intensive and require sterile techniques. Several field-applicable methods (dipslides, Petrifilm, and adenosine triphosphate [ATP] bioluminescence) were compared with the plate count for two sample matrices--phosphate-buffered saline solution containing a pure culture of Pseudomonas fluorescens and cooling-tower water containing an undefined mixed bacterial culture. For the pure culture, (1) counts determined on nutrient agar and plate-count agar (PCA) media and expressed as colony-forming units (CFU) per milliliter were equivalent to those on R2A medium (p = 1.0 and p = 1.0, respectively); (2) Petrifilm counts were not significantly different from R2A plate counts (p = 0.99); (3) the dipslide counts were up to 2 log units higher than R2A plate counts, but this discrepancy was not statistically significant (p = 0.06); and (4) a discernable correlation (r2 = 0.67) existed between ATP readings and plate counts. For cooling-tower water samples (n = 62), (1) bacterial counts using R2A medium were higher (but not significant; p = 0.63) than nutrient agar and significantly higher than tryptone-glucose yeast extract (TGE; p = 0.03) and PCA (p < 0.001); (2) Petrifilm counts were significantly lower than nutrient agar or R2A (p = 0.02 and p < 0.001, respectively), but not statistically different from TGE, PCA, and dipslides (p = 0.55, p = 0.69, and p = 0.91, respectively); (3) the dipslide method yielded bacteria counts 1 to 3 log units lower than nutrient agar and R2A (p < 0.001), but was not significantly different from Petrifilm (p = 0.91), PCA (p = 1.00) or TGE (p = 0.07); (4) the differences between dipslides and the other methods became greater with a 6-day incubation time; and (5) the correlation between ATP readings and plate counts varied from system to system, was poor (r2 values ranged from < 0.01 to 0.47), and the ATP method was not sufficiently sensitive to measure counts below approximately 10(4) CFU/mL.

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.

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.

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.

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.

Dynamics of indigenous bacterial communities associated with crude oil degradation in soil microcosms during nutrient-enhanced bioremediation.

PubMed

Chikere, Chioma B; Surridge, Karen; Okpokwasili, Gideon C; Cloete, Thomas E

2012-03-01

Bacterial population dynamics were examined during bioremediation of an African soil contaminated with Arabian light crude oil and nutrient enrichment (biostimulation). Polymerase chain reaction followed by denaturing gradient gel electrophoresis (DGGE) were used to generate bacterial community fingerprints of the different treatments employing the 16S ribosomal ribonucleic acid (rRNA) gene as molecular marker. The DGGE patterns of the nutrient-amended soils indicated the presence of distinguishable bands corresponding to the oil-contaminated-nutrient-enriched soils, which were not present in the oil-contaminated and pristine control soils. Further characterization of the dominant DGGE bands after excision, reamplification and sequencing revealed that Corynebacterium spp., Dietzia spp., Rhodococcus erythropolis sp., Nocardioides sp., Low G+C (guanine plus cytosine) Gram positive bacterial clones and several uncultured bacterial clones were the dominant bacterial groups after biostimulation. Prominent Corynebacterium sp. IC10 sequence was detected across all nutrient-amended soils but not in oil-contaminated control soil. Total heterotrophic and hydrocarbon utilizing bacterial counts increased significantly in the nutrient-amended soils 2 weeks post contamination whereas oil-contaminated and pristine control soils remained fairly stable throughout the experimental period. Gas chromatographic analysis of residual hydrocarbons in biostimulated soils showed marked attenuation of contaminants starting from the second to the sixth week after contamination whereas no significant reduction in hydrocarbon peaks were seen in the oil-contaminated control soil throughout the 6-week experimental period. Results obtained indicated that nutrient amendment of oil-contaminated soil selected and enriched the bacterial communities mainly of the Actinobacteria phylogenetic group capable of surviving in toxic contamination with concomitant biodegradation of the hydrocarbons. The present study therefore demonstrated that the soil investigated harbours hydrocarbon-degrading bacterial populations which can be biostimulated to achieve effective bioremediation of oil-contaminated soil.

Probiotic Strawberry Yogurts: Microbiological, Chemical and Sensory Properties.

PubMed

Turgut, Tamer; Cakmakci, Songül

2018-03-01

This study was performed to determine the viability of Lactobacillus acidophilus and Bifidobacterium bifidum in yogurt made with strawberry marmalade (SM) and to examine the quality properties of probiotic yogurt. Acidity, pH, bacterial counts and sensory analysis of the yogurt samples were investigated on days 1, 3, 5, 7, 10 and 14 during storage at 4 °C. The survival rate of L. acidophilus was greater than that of B. bifidum. The viability of L. acidophilus decreased during the storage period, but B. bifidum numbers remained stable during the storage period. The highest L. acidophilus count (7.20 log cfu/g) was found in L. acidophilus + B. bifidum SM yogurt on day 1. The highest B. bifidum count (6.13 log cfu/g) was detected in yogurt containing L. acidophilus + B. bifidum SM yogurt on day 7. Yeast and mould counts of all yogurts increased during the storage period. Coliform bacteria and Staphylococcus aureus were not detected in the yogurt samples. The highest overall acceptance sensory score was observed in yogurts containing L. acidophilus. Considering the sensory and probiotic characteristics of all yogurt samples, this study suggested that strawberry yogurt with a suitable 5-7 day storage period can be produced with single L. acidophilus addition or single B. bifidum addition.

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.

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.

Effect of a Benzalkonium Chloride Surfactant-Sodium Hypochlorite Combination on Elimination of Enterococcus faecalis.

PubMed

Baron, Aleksandr; Lindsey, Kimberly; Sidow, Stephanie J; Dickinson, Douglas; Chuang, Augustine; McPherson, James C

2016-01-01

The purpose of this investigation was to determine the effect of a sodium hypochlorite-surfactant combination on the removal of Enterococcus faecalis from infected teeth. Sixty-four extracted human single canal anterior teeth were prepared with rotary instrumentation and sterilized. Teeth were divided into 4 groups, N = 16. Three experimental groups were inoculated with E. faecalis and cultured for 21 days before use: positive control group, no irrigation; NaOCl group, irrigated with 5 mL 6% NaOCl; and NaOCl/BAK group, irrigated with 5 mL 6% NaOCl/0.008% benzalkonium chloride (BAK). The negative control group received medium only and no inoculate. Paper point sampling of the canals was obtained before irrigation (S1) for all 4 groups and for 2 groups after irrigation (S2) to determine remaining colony-forming units. After sampling, all teeth were split in half and evaluated for bacterial viability colony-forming units and penetration of dentinal tubules by using fluorescent vital dye staining and confocal laser scanning microscopy. Comparison of pre-irrigation and post-irrigation paper point samples from the 2 irrigated groups showed a significant reduction in bacterial canal load (P < .001, Kruskal-Wallis), with a significantly lower load in the NaOCl/BAK group than in the NaOCl group (P = .001, Mann-Whitney U test); 68.8% of the NaOCl/BAK samples gave no recoverable counts. In contrast, no significant difference between these groups was found for counts recovered from dentin. Confocal laser scanning microscopy showed no differences in tubule penetration. The addition of BAK to NaOCl significantly reduced the number of remaining bacteria within the canal after irrigation compared with NaOCl alone. Published by Elsevier Inc.

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.

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

The survival of Listeria monocytogenes during long term desiccation is facilitated by sodium chloride and organic material.

PubMed

Vogel, Birte Fonnesbech; Hansen, Lisbeth Truelstrup; Mordhorst, Hanne; Gram, Lone

2010-06-15

One specific DNA-subtype, as determined by RAPD, of Listeria monocytogenes persisted in a fish slaughterhouse for years, even during months with no production where the plant was cleaned and kept dry. We hypothesised that tolerance to desiccation could be a factor in explaining the persistence of L. monocytogenes in food processing environments and the purpose of the present study was to determine ability of L. monocytogenes to survive desiccation on stainless steel under simulated food processing conditions. Viable counts of eight different L. monocytogenes strains exposed to different soils and relative humidities (RHs) during desiccation decreased significantly (p<0.05) during the first week but subsequently remained constant at a plateau for weeks or even months thereafter. Desiccation in physiological peptone saline (PPS) reduced survivors by 3-5 log units whereas bacterial cells suspended in bacteriological growth substrates (tryptone soy broth with 1% glucose, TSB-glu) or PPS with 5% NaCl only were reduced by 1-3 log units. At RHs of 2, 43 and 75%, surfaces were visibly dry after 1, 3 and 5days of incubation, respectively. The lowest RH resulted in the most significant loss of viability, however, 10(3)-10(4)CFU/cm(2) remained viable regardless of the desiccation treatment (i.e., presence of TSB-glu and/or salt). At 75% RH, the bacterial counts remained almost constant when desiccated in TSB-glu. When bacteria were grown and desiccated (15 degrees C, 43% RH) in salmon or smoked salmon juice, survivors decreased slowly resulting in low numbers (10(2)-10(3)CFU/cm(2)) from all eight strains remaining viable after 3months. Whilst conditions during desiccation had a pronounced influence on inactivation kinetics and the number of survivors, persistent L. monocytogenes were not more tolerant to desiccation than presumed non-persistent isolates. Our study shows that the ability to survive for months during desiccated conditions may be a factor explaining the ability of L. monocytogenes to persist in food processing environments. Copyright 2010 Elsevier B.V. All rights reserved.

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.

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.

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.

Reduction in bacterial load using hypochlorous acid hygiene solution on ocular skin

PubMed Central

Stroman, David W; Mintun, Keri; Epstein, Arthur B; Brimer, Crystal M; Patel, Chirag R; Branch, James D; Najafi-Tagol, Kathryn

2017-01-01

Purpose To examine the magnitude of bacterial load reduction on the surface of the periocular skin 20 minutes after application of a saline hygiene solution containing 0.01% pure hypochlorous acid (HOCl). Methods Microbiological specimens were collected immediately prior to applying the hygiene solution and again 20 minutes later. Total microbial colonies were counted and each unique colony morphology was processed to identify the bacterial species and to determine the susceptibility profile to 15 selected antibiotics. Results Specimens were analyzed from the skin samples of 71 eyes from 36 patients. Prior to treatment, 194 unique bacterial isolates belonging to 33 different species were recovered. Twenty minutes after treatment, 138 unique bacterial isolates belonging to 26 different species were identified. Staphylococci accounted for 61% of all strains recovered and Staphylococcus epidermidis strains comprised 60% of the staphylococcal strains. No substantial differences in the distribution of Gram-positive, Gram-negative, or anaerobic species were noted before and after treatment. The quantitative data demonstrated a >99% reduction in the staphylococcal load on the surface of the skin 20 minutes following application of the hygiene solution. The total S. epidermidis colony-forming units were reduced by 99.5%. The HOCl hygiene solution removed staphylococcal isolates that were resistant to multiple antibiotics equally well as those isolates that were susceptible to antibiotics. Conclusion The application of a saline hygiene solution preserved with pure HOCl acid reduced the bacterial load significantly without altering the diversity of bacterial species remaining on the skin under the lower eyelid. PMID:28458509

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.

Duration of fever and markers of serious bacterial infection in young febrile children.

PubMed

Pratt, Amanda; Attia, Magdy W

2007-02-01

Despite the drastic change in the evaluation of the febrile young child due to the decreased incidence of serious bacterial infections (SBI) effected by Haemophilus influenza type B and pneumococcal vaccine, there remains a small role for blood work in the evaluation of these patients. Bacterial markers including white blood cell (WBC) count, absolute neutrophil count (ANC) and C-reactive protein (CRP) have been studied and are widely used as predictors of SBI in febrile children. It has been suggested that CRP values should be interpreted cautiously when fever has been present <12 h based on the kinetics of this biological marker. This limitation has not been previously addressed with CRP, nor was it described with other markers, specifically WBC and ANC, therefore the purpose of the present paper was to assess WBC, ANC and CRP values as predictors of SBI in relation to duration of fever. Patients who presented to a pediatric emergency department between the ages of 1 and 36 months, with fever > or =39 degrees C and no source of infection had a complete blood count (CBC) blood culture, and CRP level drawn. A urinalysis and/or urine culture was obtained when age and gender appropriate. A chest X-ray was performed at the discretion of the treating physician. The study subjects were enrolled prospectively and then divided into two groups based on duration of fever of < or = or >12 h, and compared. One hundred and twenty-eight patients were originally enrolled. Nine patients were excluded. Seventeen patients (14%) had SBI. One patient (<1%) had bacteremia, three (3%) had pneumonia, and 13 (10%) had urinary tract infections. Forty-five patients presented with fever < or =12 h and 74 patients presented with fever >12 h. Area under the curve (AUC) for WBC, ANC and CRP was significantly larger in patients with SBI presenting with fever >12 h (0.85, 0.83, 0.92 respectively) compared to patients with SBI who presented with fever for <12 h (0.37, 0.42, 0.68 respectively). Bacterial markers studied were more predictive of SBI if the duration of fever was >12 h as shown by the AUC. CRP performed better than WBC and ANC in both scenarios.

Intra-familial comparison of supragingival dental plaque microflora using the checkerboard DNA-DNA hybridisation technique.

PubMed

Mannaa, Alaa; Carlén, Anette; Dahlén, Gunnar; Lingström, Peter

2012-12-01

The aims of the present study were to correlate the quantified supragingival plaque bacteria between mothers and their children and identify possible microbial associations. A total of 86 mothers and their 4- to 6-year-old and 12- to 16-year-old children participated. Pooled supragingival plaque samples were obtained from interproximal sites between teeth 16/15, 25/26, 35/36 and 46/45 in mothers and older children and teeth 55/54, 64/65, 74/75 and 85/84 in younger children. All the samples were individually analysed for their content of 18 bacterial strains using checkerboard DNA-DNA hybridisation (whole genomic probes). Microbial associations were sought using cluster analysis (dendrogram) for all three age groups together, while community ordination techniques were used for each of the three groups separately. Three complexes were formed from the dendrogram in addition to associations between these complexes and remaining bacterial strains. Principal component analysis results were similar in all three groups. The correlation analyses of bacterial counts between mothers and their children showed a significant association for most of the bacterial strains (p<0.05 or 0.01). Supragingival plaque microbiota are correlated between mothers and their children. In addition, similar supragingival plaque microbial associations are present in family members.. Copyright © 2012 Elsevier Ltd. All rights reserved.

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.

Characterisation of the spoilage bacterial microbiota in oyster gills during storage at different temperatures.

PubMed

Chen, Huibin; Liu, Zhiyu; Wang, Meiying; Chen, Shaojun; Chen, Tuanwei

2013-12-01

The spoilage bacterial community in oyster gill was investigated during storage at 4, 10 and 20 °C. Aerobic plate counts and pH values were determined. Total bacterial DNA was extracted from oyster gill and bulk cells of plate count media. The major bacterial species during fresh or different temperatures storage were determined by polymerase chain reaction-denaturing gradient gel electrophoresis (PCR-DGGE). The initial aerobic plate count in oyster gill reached 6.70 log CFU g(-1). PCR-DGGE fingerprinting analysis of the 16S rRNA gene V3 region revealed that most of the strains in fresh oyster gill belonged to the genera Lactococcus and Enterobacter. The major spoilage bacteria at a storage temperature of 20 °C were Leuconostoc pseudomesenteroides, an uncultured bacterium, Cytophaga fermentans, Lactococcus lactis, Pseudoalteromonas sp., Enterococcus mundtii, Clostridium difficile and an uncultured Fusobacteria; those at 10 °C were Lactococcus spp., Lactobacillus curvatus, Weissella confusa and C. difficile; those at 4 °C were Lactococcus, Weissella, Enterobacter and Aeromonas. The other minor species were L. curvatus, Pseudomonas sp. and E. mundtii. Lactococcus spp. was the most common main spoilage bacteria in oyster gill during chilled storage. PCR-DGGE revealed the complexity of the bacterial microbiota and the major bacteria species in oyster gill for fresh and storage. © 2013 Society of Chemical Industry.

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

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

Evaluation in a Dog Model of Three Antimicrobial Glassy Coatings: Prevention of Bone Loss around Implants and Microbial Assessments.

PubMed

López-Píriz, Roberto; Solá-Linares, Eva; Rodriguez-Portugal, Mercedes; Malpica, Beatriz; Díaz-Güemes, Idoia; Enciso, Silvia; Esteban-Tejeda, Leticia; Cabal, Belén; Granizo, Juan José; Moya, José Serafín; Torrecillas, Ramón

2015-01-01

The aim of the present study is to evaluate, in a ligature-induced peri-implantitis model, the efficacy of three antimicrobial glassy coatings in the prevention of biofilm formation, intrasulcular bacterial growth and the resulting peri-implant bone loss. Mandibular premolars were bilaterally extracted from five beagle dogs. Four dental implants were inserted on each hemiarch. Eight weeks after, one control zirconia abutment and three with different bactericidal coatings (G1n-Ag, ZnO35, G3) were connected. After a plaque control period, bacterial accumulation was allowed and biofilm formation on abutments was observed by Scanning Electron Microscopy (SEM). Peri-implantitis was induced by cotton ligatures. Microbial samples and peri-implant crestal bone levels of all implant sites were obtained before, during and after the breakdown period. During experimental induce peri-implantitis: colony forming units counts from intrasulcular microbial samples at implants with G1n-Ag coated abutment remained close to the basal inoculum; G3 and ZnO35 coatings showed similar low counts; and anaerobic bacterias counts at control abutments exhibited a logarithmic increase by more than 2. Bone loss during passive breakdown period was no statistically significant. Additional bone loss occurred during ligature-induce breakdown: 0.71 (SD 0.48) at G3 coating, 0.57 (SD 0.36) at ZnO35 coating, 0.74 (SD 0.47) at G1n-Ag coating, and 1.29 (SD 0.45) at control abutments; and statistically significant differences (p<0.001) were found. The lowest bone loss at the end of the experiment was exhibited by implants dressing G3 coated abutments (mean 2.1; SD 0.42). Antimicrobial glassy coatings could be a useful tool to ward off, diminish or delay peri-implantitis progression.

Comparison of the platelet-rich plasma and buffy coat protocols for preparation of canine platelet concentrates.

PubMed

Hoareau, Guillaume L; Jandrey, Karl E; Burges, Julie; Bremer, Daphne; Tablin, Fern

2014-12-01

Platelet (PLT) concentrates (PC) can be produced via the buffy coat (BC) or platelet-rich plasma (PRP) protocols. The 2 methods have not been compared with canine blood. The aims of the study were to compare the PLT, WBC, and RBC concentrations, in vitro PLT function, and markers of platelet storage lesion (PSL) in canine PC generated by 2 different protocols, and determine microbial growth throughout storage. PC from 8 healthy donor dogs were produced using 2 standard protocols, PRP and BC. PLT, WBC, and RBC counts, optical aggregometry assays, and PSL markers (pH, pCO2 , HCO3 , lactate and glucose concentrations, and LDH activity) were determined on storage days 0, 1, 3, 5, and 7. Aerobic and anaerobic bacterial cultures were also performed. Mean PLT counts were comparable between protocols and remained stable throughout storage up to day 7, while median WBC and RBC counts on day 0 were significantly higher in the BC-PC group (17,800 WBCs/μL; 195,000 RBCs/μL) than in the PRP-PC group (200 WBCs/μL; 10,000 RBCs/μL) (P = .012). In PRP-PC aggregometry, the median slope and amplitude in response to γ-thrombin and convulxin (+ ADP) were significantly decreased, and virtually absent in BC-PC during storage. PSL markers (lactate, LDH activity) were higher in BC-PC. Aerobic bacterial growth was observed in 2 PRP-PC and 1 BC-PC. This in vitro study suggests that PRP-PC had lesser WBC and RBC contamination and superior PLT function compared with BC-PC. In vivo studies are required to address safety and efficacy of PRP-PC. © 2014 American Society for Veterinary Clinical Pathology.

Evaluating the 4-hour and 30-minute rules: effects of room temperature exposure on red blood cell quality and bacterial growth.

PubMed

Ramirez-Arcos, Sandra; Mastronardi, Cherie; Perkins, Heather; Kou, Yuntong; Turner, Tracey; Mastronardi, Emily; Hansen, Adele; Yi, Qi-Long; McLaughlin, Natasha; Kahwash, Eiad; Lin, Yulia; Acker, Jason

2013-04-01

A 30-minute rule was established to limit red blood cell (RBC) exposure to uncontrolled temperatures during storage and transportation. Also, RBC units issued for transfusion should not remain at room temperature (RT) for more than 4 hours (4-hour rule). This study was aimed at determining if single or multiple RT exposures affect RBC quality and/or promote bacterial growth. Growth and RT exposure experiments were performed in RBCs inoculated with Serratia liquefaciens and Serratia marcescens. RBCs were exposed once to RT for 5 hours (S. liquefaciens) or five times to RT for 30 minutes (S. marcescens) with periodic sampling for bacterial counts. Noncontaminated units were exposed to RT once (5 hr) or five times (30 min each) and sampled to measure in vitro quality variables. RBC core temperature was monitored using mock units with temperature loggers. Growth and RT exposure experiments were repeated three and at least six times, respectively. Statistical analysis was done using mixed-model analysis. RBC core temperature ranged from 7.3 to 11.6°C during 30-minute RT exposures and the time to reach 10°C varied from 22 to 55 minutes during 5-hour RT exposures. RBC quality was preserved after single or multiple RT exposures. Increased growth of S. liquefaciens was only observed after 2 hours of continuous RT exposure. S. marcescens concentration increased significantly in multiple-exposed units compared to the controls but did not reach clinically important levels. Single or multiple RT exposures did not affect RBC quality but slightly promoted bacterial growth in contaminated units. The clinical significance of these results remains unclear and needs further investigation. © 2012 American Association of Blood Banks.

Effect of diesel leakage in circulating cooling water system on preponderant bacteria diversity and bactericidal effect of biocides.

PubMed

Zhong, Huiyun; Liu, Fang; Lu, Jinjin; Yang, Wei; Zhao, Chaocheng

2015-01-01

Petroleum products leakage results in adverse effect on the normal operation of a circulating cooling water system. However, relatively little research has been done to explore the effect of petroleum products leakage on circulating cooling water quality and biofilm preponderant bacteria diversity. Also, normal biocides application modes cannot fulfil the need for biofilm control. In this study, diesel oil was used as the experimental subject representing leaking petroleum products; the effect of diesel addition on biofilm preponderant bacteria diversity and the bactericidal effect of chlorine dioxide and tetradecyl dimethyl benzyl ammonium chloride (1427) was investigated. Bacterial community structures were examined by PCR-denaturing gradient gel electrophoresis and PCR cloning of 16S rDNA genes. Except for 100 mg/L diesel, increasing diesel concentration enhanced the biofilm detachment ratio compared with the control test. The microstructure of biofilm samples with 0, 300 and 900 mg/L diesel addition was observed. The species of preponderant bacteria in the biofilm sample with 300 mg/L diesel addition were more and the bacterial distribution was more uniform than those in the biofilm sample with 900 mg/L diesel addition. With ClO2 and 1427 addition, chemical oxygen demand increased, lipid phosphorus and bacterial count first decreased and then remained stable, and the bactericidal ratio first increased and then remained stable. Diesel addition variation has more obvious effect on ClO2 than 1427.

Does pharmaconutrition with L-arginine and/or alpha-tocopherol improve the gut barrier in bile duct ligated rats?

PubMed

Tuncyurek, P; Sari, M; Firat, O; Mutaf, I; Gulter, C; Tunger, A; Yuce, G; Yilmaz, M; Makay, O; Dayangac, M; Ersin, S

2006-01-01

Nitric oxide supplementation and antioxidant therapy modulate gut barrier function, but the relationships between enhanced nitric oxide production, antioxidant administration, and biliary obstruction remain unclear. We evaluated the role of nitric oxide and alpha-tocopherol supplementation in bile duct ligated rats. Fifty male Wistar albino rats underwent sham operation (group I; control animals) or bile duct ligation (groups II, III, IV, and V). The ligation groups received the following regimens: standard pellet diet (group II), pellet diet plus intramuscularly administered alpha-tocopherol (group III), and L-arginine-enriched pellet diet without (group IV) or with (group V) alpha-tocopherol. Nitric oxide, malondialdehyde, and alpha-tocopherol concentrations were assessed at the end of 3 weeks. Liver and intestinal samples were scored histologically. Mesenteric lymph node and liver cultures were assessed for bacterial translocation. The liver malondialdehyde concentration was highest in group III. The nitric oxide content in the liver was higher in groups III and V, as were the blood alpha-tocopherol levels. Bacterial translocation was evident following bile duct ligation, but did not differ among the treatment groups. Intestinal histology revealed that group III had the lowest villus height, that group V had the least villus count, and that group II had the highest mucous cell count. The fibrosis scores were higher in groups IV and V. An obvious effect of alpha-tocopherol (with or without L-arginine) on the gut barrier could not be demonstrated. Moreover, the L-arginine-enriched diet promoted fibrosis in the liver. Thus, while biliary duct obstruction triggers bacterial translocation, nitric oxide and/or alpha-tocopherol supplementation did not seem to improve the gut barrier in our model. Copyright 2006 S. Karger AG, Basel.

Stable Concentrated Emulsions of the 1-Monoglyceride of Capric Acid (Monocaprin) with Microbicidal Activities against the Food-Borne Bacteria Campylobacter jejuni, Salmonella spp., and Escherichia coli

PubMed Central

Thormar, Halldor; Hilmarsson, Hilmar; Bergsson, Gudmundur

2006-01-01

Of 11 fatty acids and monoglycerides tested against Campylobacter jejuni, the 1-monoglyceride of capric acid (monocaprin) was the most active in killing the bacterium. Various monocaprin-in-water emulsions were prepared which were stable after storage at room temperature for many months and which retained their microbicidal activity. A procedure was developed to manufacture up to 500 ml of 200 mM preconcentrated emulsions of monocaprin in tap water. The concentrates were clear and remained stable for at least 12 months. They were active against C. jejuni upon 160- to 200-fold dilution in tap water and caused a >6- to 7-log10 reduction in viable bacterial count in 1 min at room temperature. The addition of 0.8% Tween 40 to the concentrates as an emulsifying agent did not change the microbicidal activity. Emulsions of monocaprin killed a variety of Campylobacter isolates from humans and poultry and also killed strains of Campylobacter coli and Campylobacter lari, indicating a broad anticampylobacter activity. Emulsions of 1.25 mM monocaprin in citrate-lactate buffer at pH 4 to 5 caused a >6- to 7-log10 reduction in viable bacterial counts of Salmonella spp. and Escherichia coli in 10 min. C. jejuni was also more susceptible to monocaprin emulsions at low pH. The addition of 5 and 10 mM monocaprin emulsions to Campylobacter-spiked chicken feed significantly reduced the bacterial contamination. These results are discussed in view of the possible utilization of monocaprin emulsions in controlling the spread of food-borne bacteria from poultry to humans. PMID:16391087

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.

Cerebrospinal fluid cytokines in the diagnosis of bacterial meningitis in infants.

PubMed

Srinivasan, Lakshmi; Kilpatrick, Laurie; Shah, Samir S; Abbasi, Soraya; Harris, Mary C

2016-10-01

Bacterial meningitis poses diagnostic challenges in infants. Antibiotic pretreatment and low bacterial density diminish cerebrospinal fluid (CSF) culture yield, while laboratory parameters do not reliably identify bacterial meningitis. Pro and anti-inflammatory cytokines are elevated in bacterial meningitis and may be useful diagnostic adjuncts when CSF cultures are negative. In a prospective cohort study of infants, we used cytometric bead arrays to measure tumor necrosis factor alpha (TNF-α), interleukin 1 (IL-1), IL-6, IL-8, IL-10, and IL-12 in CSF. Receiver operating characteristic (ROC) analyses and Principal component analysis (PCA) were used to determine cytokine combinations that identified bacterial meningitis. Six hundred and eighty four infants < 6 mo were included; 11 had culture-proven bacterial meningitis. IL-6 and IL-10 were the individual cytokines possessing greatest accuracy in diagnosis of culture proven bacterial meningitis (ROC analyses; area under the concentration-time curve (AUC) 0.91; 0.9103 respectively), and performed as well as, or better than combinations identified using ROC and PCA. CSF cytokines were highly correlated with each other and with CSF white blood cell count (WBC) counts in infants with meningitis. A subset of antibiotic pretreated culture-negative subjects demonstrated cytokine patterns similar to culture positive subjects. CSF cytokine levels may aid diagnosis of bacterial meningitis, and facilitate decision-making regarding treatment for culture negative meningitis.

The vaginal microflora in relation to gingivitis

PubMed Central

2009-01-01

Background Gingivitis has been linked to adverse pregnancy outcome (APO). Bacterial vaginosis (BV) has been associated with APO. We assessed if bacterial counts in BV is associated with gingivitis suggesting a systemic infectious susceptibilty. Methods Vaginal samples were collected from 180 women (mean age 29.4 years, SD ± 6.8, range: 18 to 46), and at least six months after delivery, and assessed by semi-quantitative DNA-DNA checkerboard hybridization assay (74 bacterial species). BV was defined by Gram stain (Nugent criteria). Gingivitis was defined as bleeding on probing at ≥ 20% of tooth sites. Results A Nugent score of 0–3 (normal vaginal microflora) was found in 83 women (46.1%), and a score of > 7 (BV) in 49 women (27.2%). Gingivitis was diagnosed in 114 women (63.3%). Women with a diagnosis of BV were more likely to have gingivitis (p = 0.01). Independent of gingival conditions, vaginal bacterial counts were higher (p < 0.001) for 38/74 species in BV+ in comparison to BV- women. Counts of four lactobacilli species were higher in BV- women (p < 0.001). Independent of BV diagnosis, women with gingivitis had higher counts of Prevotella bivia (p < 0.001), and Prevotella disiens (p < 0.001). P. bivia, P. disiens, M. curtisii and M. mulieris (all at the p < 0.01 level) were found at higher levels in the BV+/G+ group than in the BV+/G- group. The sum of bacterial load (74 species) was higher in the BV+/G+ group than in the BV+/G- group (p < 0.05). The highest odds ratio for the presence of bacteria in vaginal samples (> 1.0 × 104 cells) and a diagnosis of gingivitis was 3.9 for P. bivia (95% CI 1.5–5.7, p < 0.001) and 3.6 for P. disiens (95%CI: 1.8–7.5, p < 0.001), and a diagnosis of BV for P. bivia (odds ratio: 5.3, 95%CI: 2.6 to 10.4, p < 0.001) and P. disiens (odds ratio: 4.4, 95% CI: 2.2 to 8.8, p < 0.001). Conclusion Higher vaginal bacterial counts can be found in women with BV and gingivitis in comparison to women with BV but not gingivitis. P. bivia and P. disiens may be of specific significance in a relationship between vaginal and gingival infections. PMID:19161595

The vaginal microflora in relation to gingivitis.

PubMed

Persson, Rutger; Hitti, Jane; Verhelst, Rita; Vaneechoutte, Mario; Persson, Rigmor; Hirschi, Regula; Weibel, Marianne; Rothen, Marilynn; Temmerman, Marleen; Paul, Kathleen; Eschenbach, David

2009-01-22

Gingivitis has been linked to adverse pregnancy outcome (APO). Bacterial vaginosis (BV) has been associated with APO. We assessed if bacterial counts in BV is associated with gingivitis suggesting a systemic infectious susceptibilty. Vaginal samples were collected from 180 women (mean age 29.4 years, SD +/- 6.8, range: 18 to 46), and at least six months after delivery, and assessed by semi-quantitative DNA-DNA checkerboard hybridization assay (74 bacterial species). BV was defined by Gram stain (Nugent criteria). Gingivitis was defined as bleeding on probing at >or= 20% of tooth sites. A Nugent score of 0-3 (normal vaginal microflora) was found in 83 women (46.1%), and a score of > 7 (BV) in 49 women (27.2%). Gingivitis was diagnosed in 114 women (63.3%). Women with a diagnosis of BV were more likely to have gingivitis (p = 0.01). Independent of gingival conditions, vaginal bacterial counts were higher (p < 0.001) for 38/74 species in BV+ in comparison to BV- women. Counts of four lactobacilli species were higher in BV- women (p < 0.001). Independent of BV diagnosis, women with gingivitis had higher counts of Prevotella bivia (p < 0.001), and Prevotella disiens (p < 0.001). P. bivia, P. disiens, M. curtisii and M. mulieris (all at the p < 0.01 level) were found at higher levels in the BV+/G+ group than in the BV+/G- group. The sum of bacterial load (74 species) was higher in the BV+/G+ group than in the BV+/G- group (p < 0.05). The highest odds ratio for the presence of bacteria in vaginal samples (> 1.0 x 104 cells) and a diagnosis of gingivitis was 3.9 for P. bivia (95% CI 1.5-5.7, p < 0.001) and 3.6 for P. disiens (95%CI: 1.8-7.5, p < 0.001), and a diagnosis of BV for P. bivia (odds ratio: 5.3, 95%CI: 2.6 to 10.4, p < 0.001) and P. disiens (odds ratio: 4.4, 95% CI: 2.2 to 8.8, p < 0.001). Higher vaginal bacterial counts can be found in women with BV and gingivitis in comparison to women with BV but not gingivitis. P. bivia and P. disiens may be of specific significance in a relationship between vaginal and gingival infections.

[Factors affecting the DAPI fluorescence direct count in the tidal river sediment].

PubMed

Chen, Chen; Huang, Shan; Wu, Qun-he; Li, Rui-yi; Zhang, Ren-duo

2010-08-01

The factors affecting the DAPI (4', 6-diamidino-2-phenylidole) fluorescence direct count in the tidal river sediment were examined. Sediment samples were collected from the Guangzhou section of the Pearl River. Besides sediment texture and organic matter, an improved staining procedure and the involved parameters were analyzed. Results showed that the procedure with the sediment with 2000 fold dilution and ultrasonic water bath for 10 min, and with a final DAPI concentration of 10 microg x mL(-1) and staining time for more than 30 min produced the optimum results of DAPI direct count in the sediment. The total bacterial number was correlated to the proportion of the non-nucleoid-containing cells to the total bacterial number (r = 0.587, p = 0.004). The organic matter content also correlated to the ration. The clay content had a strong correlation with the organic matter, through which the clay content also affected the ratio. A multiple regression analysis between the ration versus the organic matter, the total bacterial number, and the clay content showed that the regression equation fit the measure values satisfactorily (r = 0.694). These results indicated that the above factors needed to be considered in the applications of the DAPI fluorescence direct counting method to the tidal river sediment.

Superiority of a functional leukocyte adhesiveness/aggregation test over the white blood cell count to discriminate between mild and significant inflammatory response in patients with acute bacterial infections.

PubMed

Rogowski, Ori; Rotstein, Rivka; Zeltzer, David; Misgav, Sarit; Justo, Daniel; Avitzour, Daniel; Mardi, Tamar; Serov, Jacob; Arber, Nadir; Berliner, Shlomo; Shapira, Itzhak

2002-01-01

Electronic cell counters may underestimate the white blood cell count (WBCC) in the presence of aggregated leukocytes. In the present study we focused on the possibility of using a functional, as opposed to an anatomic, count to circumvent this eventual underestimation. A model of bacterial infection was used because of the importance of leukocytosis in the physician's clinical decision-making process. There were 35 patients with low C-reactive protein (CRP) concentrations (0.5-4.9 mg/dL), 45 with intermediate (5-9.9 mg/dL), and 120 with relatively high (>10 mg/dL) CRP concentrations. A significant (P=0.008) difference was noted between the state of leukocyte adhesiveness/aggregation in the peripheral blood of individuals with low CRP concentrations (3.5%+/-4.3%) and those with high CRP concentrations (7.4%+/-8%), while there was no significant difference in the respective number of WBCs per cubic millimeter (cmm) (11,600 +/- 5,500 and 14,000 +/- 7,200, respectively). We raise the possibility that a functional test might be superior over an anatomic count in patients with acute bacterial infection and a significant acute phase response. Copyright 2002 Wiley-Liss, Inc.

Effect of Probiotic Lactobacillus reuteri on Salivary Cariogenic Bacterial Counts among Groups of Preschool Children in Jeddah, Saudi Arabia: A Randomized Clinical Trial.

PubMed

Alamoudi, Najlaa M; Almabadi, Eman S; El Ashiry, Eman A; El Derwi, Douaa A

2018-05-15

To evaluate the effect of probiotic Lactobacilli reuteri lozenges on caries-associated salivary bacterial counts (Mutans streptococci and Lactobacillus), dental plaque accumulation, and salivary buffer capacity in a group of preschool children. The study group consisted of 178 healthy children (aged 3-6 years). Children were randomly grouped: the experimental group (n = 90) received L. reuteri probiotic lozenges and the control group (n = 88) received placebo lozenges, twice daily, for 28 days. Salivary Mutans streptococci and Lactobacillus counts, and buffer capacity were assessed using chair-side caries-risk test (CRT®) kits. The Simplified Oral Hygiene index (OHI-S) was used to assess dental plaque accumulation at baseline and after 28 days. After 28 days, the experimental group had a statistically significant reduction in Mutans streptococci and lactobacilli (p = 0.000 and p = 0.020, respectively) and both groups had less plaque accumulation than at baseline. While the buffer capacity in the experimental group increased more than in the control group, it was not statistically significant (p = 0.577). Compliance was 90%, with no adverse events. Consumption of probiotic lozenges containing L. reuteri reduces caries-associated bacterial counts significantly. Probiotics consumption may have a beneficial caries-preventive effect.

Diagnostic markers of serious bacterial infections in febrile infants younger than 90 days old.

PubMed

Nosrati, Adi; Ben Tov, Amir; Reif, Shimon

2014-02-01

The aim of this study was to assess correlations between demographic, clinical and laboratory characteristics and the risk of serious bacterial infection (SBI) in febrile <90-day-old infants. Medical records of all infants younger than 90 days old hospitalized at Dana-Dwek Children's Hospital (2006-2008) for evaluation of fever were retrospectively reviewed. Data on clinical, laboratory and demographic characteristics were retrieved and evaluated. Forty-eight of the 401 study infants (12%) had SBI: most of them had urinary tract infection (43 infants; 90% of all SBI), three infants had bacteremia, one had bacterial pneumonia and one had bacterial meningitis. Significant independent clinical predictors for the diagnosis of SBI included duration of fever, absence of rhinitis and the absence of lung and skin manifestations. Significant independent laboratory predictors were absolute neutrophil count (ANC), platelets, blood urea nitrogen and C-reactive protein (CRP) level. On receiver operating characteristic curve analysis, the CRP area under the curve (0.819) was significantly superior to ANC and leukocyte count. Of the clinical and laboratory variables selected for evaluation, qualitative CRP was the strongest independent predictor for diagnosing SBI and a significantly better diagnostic marker than clinical characteristics, ANC and white blood cell count. © 2013 The Authors. Pediatrics International © 2013 Japan Pediatric Society.

Immunological and infectious risk factors for lung cancer in US veterans with HIV: a longitudinal cohort study.

PubMed

Sigel, Keith; Wisnivesky, Juan; Crothers, Kristina; Gordon, Kirsha; Brown, Sheldon T; Rimland, David; Rodriguez-Barradas, Maria C; Gibert, Cynthia; Goetz, Matthew Bidwell; Bedimo, Roger; Park, Lesley S; Dubrow, Robert

2017-02-01

HIV infection is independently associated with risk of lung cancer, but few data exist for the relation between longitudinal measurements of immune function and lung-cancer risk in people living with HIV. We followed up participants with HIV from the Veterans Aging Cohort Study for a minimum of 3 years between Jan 1, 1998, and Dec 31, 2012, and used cancer registry data to identify incident cases of lung cancer. The index date for each patient was the later of the date HIV care began or Jan 1, 1998. We excluded patients with less than 3 years' follow-up, prevalent diagnoses of lung cancer, or incomplete laboratory data. We used Cox regression models to investigate the relation between different time-updated lagged and cumulative exposures (CD4 cell count, CD8 cell count, CD4/CD8 ratio, HIV RNA, and bacterial pneumonia) and risk of lung cancer. Models were adjusted for age, race or ethnicity, smoking, hepatitis C virus infection, alcohol use disorders, drug use disorders, and history of chronic obstructive pulmonary disease and occupational lung disease. We identified 277 cases of incident lung cancer in 21 666 participants with HIV. In separate models for each time-updated 12 month lagged, 24 month simple moving average cumulative exposure, increased risk of lung cancer was associated with low CD4 cell count (p trend=0·001), low CD4/CD8 ratio (p trend=0·0001), high HIV RNA concentration (p=0·004), and more cumulative bacterial pneumonia episodes (12 month lag only; p trend=0·0004). In a mutually adjusted model including these factors, CD4/CD8 ratio and cumulative bacterial pneumonia episodes remained significant (p trends 0·003 and 0·004, respectively). In our large HIV cohort in the antiretroviral therapy era, we found evidence that dysfunctional immune activation and chronic inflammation contribute to the development of lung cancer in the setting of HIV infection. These findings could be used to target lung-cancer prevention measures to high-risk groups. US National Institutes of Health. Copyright © 2017 Elsevier Ltd. All rights reserved.

Microbial Contamination on Used Surgical Masks among Hospital Personnel and Microbial Air Quality in their Working Wards: A Hospital in Bangkok

PubMed Central

Luksamijarulkul, Pipat; Aiempradit, Natkitta; Vatanasomboon, Pisit

2014-01-01

Objective To assess the relationship of bacterial and fungal contamination on used surgical masks worn by the hospital personnel and microbial air quality in their working wards. Methods This is a cross-sectional study of 230 used surgical masks collected from 214 hospital personnel, and 215 indoor air samples collected from their working wards to culture for bacterial and fungal counts. This study was carried out at the hospital in Bangkok. Group or genus of isolated bacteria and fungi were preliminarily identified by Gram’s stain and lacto-phenol cotton blue. Data were analyzed using paired t-test and Pearson’s correlation coefficient at the significant level of p<0.050. Results Means and standard deviation of bacterial and fungal contamination on inside area of the used masks were 47 ± 56 and 15 ± 9 cfu/ml/piece, and on outside area were 166 ± 199 and 34 ± 18 cfu/ml/piece, respectively, p<0.001. The bacterial and fungal contamination on used masks from hospital personnel working in the male and female medical wards and out-patient department, as well as the bacterial and fungal counts of the indoor air sample collected from the same area were relatively higher than the other wards. The predominant isolated bacteria and fungi contaminated on inside and outside areas of the used masks and air samples were similar (Staphylococcus spp. and Aspergillus spp.; respectively). For its relationship, results found that bacterial and fungal counts in air samples showed significantly positive correlation with the bacterial contamination load on outside area of the used masks, r=0.16, p=0.018 and r=0.21, p=0.003, respectively. Conclusion High bacterial contamination on outside area of the used masks was demonstrated, and it showed a significant correlation with microbial air quality of working wards. PMID:25337311

Microbial Contamination on Used Surgical Masks among Hospital Personnel and Microbial Air Quality in their Working Wards: A Hospital in Bangkok.

PubMed

Luksamijarulkul, Pipat; Aiempradit, Natkitta; Vatanasomboon, Pisit

2014-09-01

To assess the relationship of bacterial and fungal contamination on used surgical masks worn by the hospital personnel and microbial air quality in their working wards. This is a cross-sectional study of 230 used surgical masks collected from 214 hospital personnel, and 215 indoor air samples collected from their working wards to culture for bacterial and fungal counts. This study was carried out at the hospital in Bangkok. Group or genus of isolated bacteria and fungi were preliminarily identified by Gram's stain and lacto-phenol cotton blue. Data were analyzed using paired t-test and Pearson's correlation coefficient at the significant level of p<0.050. Means and standard deviation of bacterial and fungal contamination on inside area of the used masks were 47 ± 56 and 15 ± 9 cfu/ml/piece, and on outside area were 166 ± 199 and 34 ± 18 cfu/ml/piece, respectively, p<0.001. The bacterial and fungal contamination on used masks from hospital personnel working in the male and female medical wards and out-patient department, as well as the bacterial and fungal counts of the indoor air sample collected from the same area were relatively higher than the other wards. The predominant isolated bacteria and fungi contaminated on inside and outside areas of the used masks and air samples were similar (Staphylococcus spp. and Aspergillus spp.; respectively). For its relationship, results found that bacterial and fungal counts in air samples showed significantly positive correlation with the bacterial contamination load on outside area of the used masks, r=0.16, p=0.018 and r=0.21, p=0.003, respectively. High bacterial contamination on outside area of the used masks was demonstrated, and it showed a significant correlation with microbial air quality of working wards.

SPARTA: Simple Program for Automated reference-based bacterial RNA-seq Transcriptome Analysis.

PubMed

Johnson, Benjamin K; Scholz, Matthew B; Teal, Tracy K; Abramovitch, Robert B

2016-02-04

Many tools exist in the analysis of bacterial RNA sequencing (RNA-seq) transcriptional profiling experiments to identify differentially expressed genes between experimental conditions. Generally, the workflow includes quality control of reads, mapping to a reference, counting transcript abundance, and statistical tests for differentially expressed genes. In spite of the numerous tools developed for each component of an RNA-seq analysis workflow, easy-to-use bacterially oriented workflow applications to combine multiple tools and automate the process are lacking. With many tools to choose from for each step, the task of identifying a specific tool, adapting the input/output options to the specific use-case, and integrating the tools into a coherent analysis pipeline is not a trivial endeavor, particularly for microbiologists with limited bioinformatics experience. To make bacterial RNA-seq data analysis more accessible, we developed a Simple Program for Automated reference-based bacterial RNA-seq Transcriptome Analysis (SPARTA). SPARTA is a reference-based bacterial RNA-seq analysis workflow application for single-end Illumina reads. SPARTA is turnkey software that simplifies the process of analyzing RNA-seq data sets, making bacterial RNA-seq analysis a routine process that can be undertaken on a personal computer or in the classroom. The easy-to-install, complete workflow processes whole transcriptome shotgun sequencing data files by trimming reads and removing adapters, mapping reads to a reference, counting gene features, calculating differential gene expression, and, importantly, checking for potential batch effects within the data set. SPARTA outputs quality analysis reports, gene feature counts and differential gene expression tables and scatterplots. SPARTA provides an easy-to-use bacterial RNA-seq transcriptional profiling workflow to identify differentially expressed genes between experimental conditions. This software will enable microbiologists with limited bioinformatics experience to analyze their data and integrate next generation sequencing (NGS) technologies into the classroom. The SPARTA software and tutorial are available at sparta.readthedocs.org.

Studies on stannous fluoride toothpaste and gel (2). Effects on salivary bacterial counts and plaque regrowth in vivo.

PubMed

Addy, M; Greenman, J; Renton-Harper, P; Newcombe, R; Doherty, F

1997-02-01

There has been a resurgence of interest in stannous fluoride (SF) products in particular to provide oral hygiene and gingival health benefits. The aim of this study was to assess the persistence of antimicrobial action of a number of SF formulations in the mouth and relate these to plaque inhibitory activity. The formulations were 2 SF toothpastes (SF1, SF2), 2 SF plus stannous pyrophosphate toothpastes (SFSP1, SFSP2), a SF gel (G), a NaF toothpaste (C) and saline (S) as control. Both studies involve 2 different groups of 21 healthy dentate volunteers. The studies were single, blind, randomised, crossover designs balanced for residual effects, with a minimum 2 1/2 day washout period. Salivary bacterial counts were determined before and to 7 h after a single rinse with the formulations. Plaque regrowth from a zero baseline (day 1) was measured by index and area on day 5, after 2x daily rinsing with slurries of the formulations or saline. For bacterial counts, highly significant treatment differences were found. Bacterial counts were variably reduced by all treatments to 30 min then showed a variable rate of return towards baseline. All test agents were significantly better than S at some timepoints. The order for greatest persistence of action downwards was; (1) SFSP2; (2) SFSP1, G, and SF1; (3) SF2; (4) C; (5) S. Highly significant differences in plaque regrowth between treatments were found with similar mean ordering of efficacy as for salivary bacterial counts from most effective downwards namely; (1) SFSP1 and SFSP2; (2) SF1; (3) SF2; G and C; (4) S. The results were consistent with a parallel study measuring tea staining in vitro, whereby formulations causing the most staining produced the greatest persistence of action and plaque inhibitory activity. This suggests the availability of stannous ions was important for the clinical effects. It is concluded that stannous ions can enhance the plaque inhibitory action of toothpaste via a persistent antimicrobial action.

Devitalization of bacterial and parasitic germs in sewage sludge during aerobic digestion under laboratory conditions.

PubMed

Juris, P; Plachý, P; Lauková, A

1995-05-01

The survival of 8 bacterial species (Pseudomonas sp., Salmonella sp., Enterobacteriae, Streptococcus sp., Escherichia coli) was detected in municipal sewage sludge up to 37 hours of mesophilic aerobic digestion under laboratory conditions. The model strain Enterococcus faecium CCM 4231 survived almost twice as long as the above-mentioned isolates. Similar findings, regarding the viability of the microorganisms studied, were also determined during thermophilic aerobic digestion of municipal sewage sludges. The final reduction in the total count of bacteria was not directly dependent on the temperature during aerobic digestion. It may be supposed that E. faecium CCM 4231 strain as a bacteriocin-producing strain with a broad antimicrobial spectrum, inoculated into the sludges, could inhibit the growth of microorganisms in the sludges by the way of its bacteriocin activity. Studying the effect of aerobic digestion on the viability of helminth eggs, the observed negative effect of higher temperatures was more expressive in comparison with bacterial strains. During thermophilic digestion process all helminth eggs (Ascaris suum, Toxocara canis) were devitalized. All eggs of T. canis were killed in experiments under mesophilic temperature. However, 32% of nonembryonated A. suum eggs remained viable.

Protection from Staphylococcus aureus mastitis associated with poly-N-acetyl β-1,6 glucosamine specific antibody production using biofilm-embedded bacteria

PubMed Central

Pérez, M. M.; Prenafeta, A.; Valle, J.; Penadés, J.; Rota, C.; Solano, C.; Marco, J.; Grilló, M.J.; Lasa, I.; Irache, J.M.; Maira-Litran, T.; Jiménez-Barbero, J.; Costa, L.; Pier, G.B.; de Andrés, D.; Amorena, B.

2010-01-01

Staphylococcus aureus vaccines based on bacterins surrounded by slime, surface polysaccharides coupled to protein carriers and polysaccharides embedded in liposomes administered together with non-biofilm bacterins confer protection against mastitis. However, it remains unknown whether protective antibodies are directed to slime-associated known exopolysaccharides and could be produced in the absence of bacterin immunizations. Here, a sheep mastitis vaccination study was carried out using bacterins, crude bacterial extracts or a purified exopolysaccharide from biofilm bacteria delivered in different vehicles. This polysaccharide reacted specifically with antibodies to poly-N-acetyl-β-1,6-glucosamine (PNAG) and not with antibodies to other capsular antigens or bacterial components. Following intra-mammary challenge with biofilm-producing bacteria, antibody production against the polysaccharide, milk bacterial counts and mastitis lesions were determined. Bacterins from strong biofilm-producing bacteria triggered the highest production of antibodies to PNAG and conferred the highest protection against infection and mastitis, compared with weak biofilm-producing bacteria and non-cellular inocula. Thus, bacterins from strong biofilm bacteria, rather than purified polysaccharide, are proposed as a cost-efficient vaccination against S. aureus ruminant mastitis. PMID:19428854

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.

Do thawing and warming affect the integrity of human milk?

PubMed

Handa, D; Ahrabi, A F; Codipilly, C N; Shah, S; Ruff, S; Potak, D; Williams, J E; McGuire, M A; Schanler, R J

2014-11-01

To evaluate the integrity of the human milk (pH, bacterial counts, host defense factors and nutrients) subjected to thawing, warming, refrigeration and maintenance at room temperature. Mothers in the neonatal intensive care unit donated freshly expressed milk. A baseline sample was stored at -80 °C and the remainder of the milk was divided and stored for 7 days at -20 °C. The milk was then subjected to two methods of thawing and warming: tepid water and waterless warmer. Thawed milk also was refrigerated for 24 h prior to warming. Lastly, warmed milk was maintained at room temperature for 4 h to simulate a feeding session. Samples were analyzed for pH, bacterial colony counts, total fat and free fatty acids, and the content of protein, secretory IgA and lactoferrin. Data were analyzed by repeated-measures analysis of variance and paired t test. There were no differences between processing methods and no changes in fat, protein, lactoferrin and secretory immunoglobulin A with processing steps. Milk pH and bacterial colony counts declined while free fatty acids rose with processing. Refrigeration of thawed milk resulted in greater declines in pH and bacteria and increases in free fatty acids. Bacterial colony counts and free fatty acids increased with maintenance at room temperature. The integrity of the milk was affected similarly by the two thawing and warming methods. Thawing and warming change the integrity of previously frozen human milk, but not adversely. Concerns about maintaining warmed milk at room temperature need to be explored.

Evaluation of changes in periodontal bacteria in healthy dogs over 6 months using quantitative real-time PCR.

PubMed

Maruyama, N; Mori, A; Shono, S; Oda, H; Sako, T

2018-03-01

Porphyromonas gulae, Tannerella forsythia and Campylobacter rectus are considered dominant periodontal pathogens in dogs. Recently, quantitative real-time PCR (qRT-PCR) methods have been used for absolute quantitative determination of oral bacterial counts. The purpose of the present study was to establish a standardized qRT-PCR procedure to quantify bacterial counts of the three target periodontal bacteria (P. gulae, T. forsythia and C. rectus). Copy numbers of the three target periodontal bacteria were evaluated in 26 healthy dogs. Then, changes in bacterial counts of the three target periodontal bacteria were evaluated for 24 weeks in 7 healthy dogs after periodontal scaling. Analytical evaluation of each self-designed primer indicated acceptable analytical imprecision. All 26 healthy dogs were found to be positive for P. gulae, T. forsythia and C. rectus. Median total bacterial counts (copies/ng) of each target genes were 385.612 for P. gulae, 25.109 for T. forsythia and 5.771 for C. rectus. Significant differences were observed between the copy numbers of the three target periodontal bacteria. Periodontal scaling reduced median copy numbers of the three target periodontal bacteria in 7 healthy dogs. However, after periodontal scaling, copy numbers of all three periodontal bacteria significantly increased over time (p<0.05, Kruskal-Wallis test) (24 weeks). In conclusion, our results demonstrated that qRT-PCR can accurately measure periodontal bacteria in dogs. Furthermore, the present study has revealed that qRT-PCR method can be considered as a new objective evaluation system for canine periodontal disease. Copyright© by the Polish Academy of Sciences.

Preponderance of bacterial isolates in urine of HIV-positive malaria-infected pregnant women with urinary tract infection.

PubMed

Ako-Nai, Kwashie Ajibade; Ebhodaghe, Blessing Itohan; Osho, Patrick; Adejuyigbe, Ebun; Adeyemi, Folasade Mubiat; Kassim, Olakunle O

2014-12-15

This study examined HIV and malaria co-infection as a risk factor for urinary tract infections (UTIs) in pregnancy. The study group included 74 pregnant women, 20 to 42 years of age, who attended the antenatal clinic at the Specialist Hospital at Akure, Ondo State, Nigeria. Forty-four of the pregnant women were either HIV seropositive with malaria infection (HIV+Mal+) or HIV seropositive without malaria (HIV+Mal-). The remaining thirty pregnant women served as controls and included women HIV seronegative but with malaria (HIV-Mal+) and women HIV seronegative without malaria. UTI was indicated by a bacterial colony count of greater than 10⁵/mL of urine, using cysteine lactose electrolyte deficient medium (CLED) as the primary isolation medium. Bacterial isolates were characterized using convectional bacteriological methods, and antibiotics sensitivity tests were carried out using the disk diffusion method. A total of 246 bacterial isolates were recovered from the cultures, with a mean of 3.53 isolates per subject. Women who were HIV+Mal+ had the most diverse group of bacterial isolates and the highest frequency of UTIs. The bacterial isolates from the HIV+Mal+ women also showed the highest degree of antibiotic resistance. While pregnancy and HIV infection may each represent a risk factor for UTI, HIV and malaria co-infection may increase its frequency in pregnancy. The higher frequency of multiple antibiotic resistance observed among the isolates, particularly isolates from HIV+Mal+ subjects, poses a serious public health concern as these strains may aggravate the prognosis of both UTI and HIV infection.

Inoculation of a phenanthrene-degrading endophytic bacterium reduces the phenanthrene level and alters the bacterial community structure in wheat.

PubMed

Liu, Juan; Xiang, Yanbing; Zhang, Zhiming; Ling, Wanting; Gao, Yanzheng

2017-06-01

Colonization by polycyclic aromatic hydrocarbon (PAH)-degrading endophytic bacteria (PAHDEB) can reduce the PAH contamination risk in plant. However, little information is available on the impact of PAHDEB colonization on the endophytic bacterial community of inner plant tissues. A phenanthrene-degrading endophytic bacterium (PDEB), Massilia sp. Pn2, was inoculated onto the roots of wheat and subjected to greenhouse container experiments. The endophytic bacterial community structure in wheat was investigated using high-throughput sequencing technology. The majority of endophytic bacteria in wheat were Proteobacteria, and the dominant genus was Pseudomonas. Phenanthrene contamination clearly increased the diversity of endophytic bacteria in wheat. The cultivable endophytic bacteria counts in wheat decreased with increasing the level of phenanthrene contamination; the endophytic bacterial community structure changed correspondingly, and the bacterial richness first increased and then decreased. Inoculation of strain Pn2 reduced the phenanthrene contamination in wheat, enlarged the biomass of wheat roots, changed the bacterial community structure and enhanced the cell counts, diversity and richness of endophytic bacteria in phenanthrene-contaminated wheat in a contamination level-dependent manner. The findings of this investigation provide insight into the responses of endophytic bacterial community in plant to external PAH contamination and PAHDEB colonization.

International Space Station environmental microbiome - microbial inventories of ISS filter debris.

PubMed

Venkateswaran, Kasthuri; Vaishampayan, Parag; Cisneros, Jessica; Pierson, Duane L; Rogers, Scott O; Perry, Jay

2014-01-01

Despite an expanding array of molecular approaches for detecting microorganisms in a given sample, rapid and robust means of assessing the differential viability of the microbial cells, as a function of phylogenetic lineage, remain elusive. A propidium monoazide (PMA) treatment coupled with downstream quantitative polymerase chain reaction (qPCR) and pyrosequencing analyses was carried out to better understand the frequency, diversity, and distribution of viable microorganisms associated with debris collected from the crew quarters of the International Space Station (ISS). The cultured bacterial counts were more in the ISS samples than cultured fungal population. The rapid molecular analyses targeted to estimate viable population exhibited 5-fold increase in bacterial (qPCR-PMA assay) and 25-fold increase in microbial (adenosine triphosphate assay) burden than the cultured bacterial population. The ribosomal nucleic acid-based identification of cultivated strains revealed the presence of only four to eight bacterial species in the ISS samples, however, the viable bacterial diversity detected by the PMA-pyrosequencing method was far more diverse (12 to 23 bacterial taxa) with the majority consisting of members of actinobacterial genera (Propionibacterium, Corynebacterium) and Staphylococcus. Sample fractions not treated with PMA (inclusive of both live and dead cells) yielded a great abundance of highly diverse bacterial (94 to 118 taxa) and fungal lineages (41 taxa). Even though deep sequencing capability of the molecular analysis widened the understanding about the microbial diversity, the cultivation assay also proved to be essential since some of the spore-forming microorganisms were detected only by the culture-based method. Presented here are the findings of the first comprehensive effort to assess the viability of microbial cells associated with ISS surfaces, and correlate differential viability with phylogenetic affiliation.

Effect of early antibiotic administration on cecal bacterial communities and their metabolic profiles in pigs fed diets with different protein levels.

PubMed

Zhang, Chuanjian; Yu, Miao; Yang, Yuxiang; Mu, Chunlong; Su, Yong; Zhu, Weiyun

2016-12-01

This study investigated the effects of early antibiotic administration (EAA) on cecal bacterial communities and their metabolic profiles in pigs fed diets with different protein levels. Eighteen litters (total 180) of piglets on day (d) 7 were fed either a commercial creep feed or commercial creep feed + antibiotic (Olaquindox, Oxytetracycline Calcium and Kitasamycin) until d 42. On d 42, pigs within each group were further randomly fed a normal crude protein (CP) diet (20% and 18% CP from d 42 to d 77 and d 77 to d 120, respectively) or a low-CP diet (16% and 14% CP from d 42 to d 77 and d 77 to d 120, respectively), generating 4 groups, control-low CP (Con-LP), control-normal CP (Con-NP), antibiotic-low CP (Ant-LP) and antibiotic-normal CP (Ant-NP), respectively. On d 77 and d 120, 5 pigs per group were slaughtered and cecal materials were collected for bacterial analysis. With cecal bacteria, principle component analysis (PCA) of the denaturing gradient gel electrophoresis (DGGE) profile showed two distinct groups of samples from low-CP diet and samples from normal-CP diet. Real-time PCR showed that EAA did not have significant effect on major bacterial groups, only showed significant interactions (P < 0.05) with CP level for Lactobacillus counts on d 77 and Clostridium cluster XIVa counts on d 120 with higher values in the Con-NP group compared to the Ant-NP groups. Low-CP diet increased (P < 0.05) short-chain fatty acids (SCFA) producing bacteria counts (Bacteroidetes on d 77 and d 120; Clostridium cluster IV and Clostridium cluster XIVa on d 77), but decreased (P < 0.05) Escherichia coli counts on d 77 and d 120. For metabolites, EAA increased (P < 0.05) protein fermentation products (p-cresol, indole and skatole on d 77; ammonia, putrescine and spermidine on d 120), and showed significant interactions (P < 0.05) with CP level for p-cresol and skatole concentrations on d 77 and putrescine and spermidine concentrations on d 120 with higher values in the Ant-LP group compared to the Con-LP groups. Low-CP diet increased (P < 0.05) SCFA concentration (propionate and butyrate) on d 77, but reduced (P < 0.05) the protein fermentation products (ammonia, phenol and indole on d 77; branched chain fatty acid (BCFA), ammonia, tyramine, cadaverine and indole on d 120). These results indicate that EAA had less effect on bacterial communities, but increased bacterial fermentation of protein in the cecum under low-CP diet. Low-CP diet altered bacterial communities with an increase in the counts of SCFA-producing bacteria and a decrease in the counts of Escherichia coli, and markedly reduced the protein fermentation products. Copyright © 2016 Elsevier Ltd. All rights reserved.

Is ultraviolet radiation on haemodialysis RO water beneficial?

PubMed

Stragier, A

2005-01-01

The quality of dialysis fluids has become increasingly important in the treatment of HD patients. Purified water represents over 95% of its volume. Bacterial and endotoxin content of Reverse Osmosis (RO) water is usually kept under control by bacterial filters, inserted in the distribution departure loop, and by monthly disinfection of the distribution circuit; the simpler the circuit, the better. This paper reports 12 years experience during which Ultraviolet Irradiation (UV) has replaced bacterial filters. To keep the bacterial growth under control in a complex RO water circuit (including a tank and multiple loops) a simple UV lamp was inserted in the departure line. It proved sufficient to keep bacterial count within AAMI norms. Failure of the UV lamp was associated with a rise of up to 500 cfu/ml in the last (fourth week) before routine disinfection. Normal levels were again obtained after replacement of the UV lamp. Six years later, a second UV lamp was added on the return loop. Bacterial counts and endotoxin levels in RO water promptly fell to <1 cfu/ml and <0.125 EU, till today. It is concluded that UV lamps should be favoured over bacterial filters in systems that are not disinfected daily, such as the RO water circuit. The principle of UV irradiation is explained and its advantage over bacterial filters is discussed. Future possible applications of UV are presented.

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.

Evaluation of Petrifilm Lactic Acid Bacteria Plates for Counting Lactic Acid Bacteria in Food.

PubMed

Kanagawa, Satomi; Ohshima, Chihiro; Takahashi, Hajime; Burenqiqige; Kikuchi, Misato; Sato, Fumina; Nakamura, Ayaka; Mohamed, Shimaa M; Kuda, Takashi; Kimura, Bon

2018-06-01

Although lactic acid bacteria (LAB) are used widely as starter cultures in the production of fermented foods, they are also responsible for food decay and deterioration. The undesirable growth of LAB in food causes spoilage, discoloration, and slime formation. Because of these adverse effects, food companies test for the presence of LAB in production areas and processed foods and consistently monitor the behavior of these bacteria. The 3M Petrifilm LAB Count Plates have recently been launched as a time-saving and simple-to-use plate designed for detecting and quantifying LAB. This study compares the abilities of Petrifilm LAB Count Plates and the de Man Rogosa Sharpe (MRS) agar medium to determine the LAB count in a variety of foods and swab samples collected from a food production area. Bacterial strains isolated from Petrifilm LAB Count Plates were identified by 16S rDNA sequence analysis to confirm the specificity of these plates for LAB. The results showed no significant difference in bacterial counts measured by using Petrifilm LAB Count Plates and MRS medium. Furthermore, all colonies growing on Petrifilm LAB Count Plates were confirmed to be LAB, while yeast colonies also formed in MRS medium. Petrifilm LAB Count Plates eliminated the plate preparation and plate inoculation steps, and the cultures could be started as soon as a diluted food sample was available. Food companies are required to establish quality controls and perform tests to check the quality of food products; the use of Petrifilm LAB Count Plates can simplify this testing process for food companies.

NOWCASTING AND FORECASTING BEACH BACTERIA CONCENTRATION USING THE EPA VIRTUAL BEACH SOFTWARE

EPA Science Inventory

Beaches are subject to closure when bacterial counts exceed water quality criteria. Many authorities base these decisions on sample counts, which typically require a day or more to analyze. Sometimes called the persistence model, because conditions are assumed to persist, experie...

Variation in Honey Bee Gut Microbial Diversity Affected by Ontogenetic Stage, Age and Geographic Location

PubMed Central

Hroncova, Zuzana; Havlik, Jaroslav; Killer, Jiri; Doskocil, Ivo; Tyl, Jan; Kamler, Martin; Titera, Dalibor; Hakl, Josef; Mrazek, Jakub; Bunesova, Vera; Rada, Vojtech

2015-01-01

Social honey bees, Apis mellifera, host a set of distinct microbiota, which is similar across the continents and various honey bee species. Some of these bacteria, such as lactobacilli, have been linked to immunity and defence against pathogens. Pathogen defence is crucial, particularly in larval stages, as many pathogens affect the brood. However, information on larval microbiota is conflicting. Seven developmental stages and drones were sampled from 3 colonies at each of the 4 geographic locations of A. mellifera carnica, and the samples were maintained separately for analysis. We analysed the variation and abundance of important bacterial groups and taxa in the collected bees. Major bacterial groups were evaluated over the entire life of honey bee individuals, where digestive tracts of same aged bees were sampled in the course of time. The results showed that the microbial tract of 6-day-old 5th instar larvae were nearly equally rich in total microbial counts per total digestive tract weight as foraging bees, showing a high percentage of various lactobacilli (Firmicutes) and Gilliamella apicola (Gammaproteobacteria 1). However, during pupation, microbial counts were significantly reduced but recovered quickly by 6 days post-emergence. Between emergence and day 6, imago reached the highest counts of Firmicutes and Gammaproteobacteria, which then gradually declined with bee age. Redundancy analysis conducted using denaturing gradient gel electrophoresis identified bacterial species that were characteristic of each developmental stage. The results suggest that 3-day 4th instar larvae contain low microbial counts that increase 2-fold by day 6 and then decrease during pupation. Microbial succession of the imago begins soon after emergence. We found that bacterial counts do not show only yearly cycles within a colony, but vary on the individual level. Sampling and pooling adult bees or 6th day larvae may lead to high errors and variability, as both of these stages may be undergoing dynamic succession. PMID:25768309

Comparison of the test characteristics of procalcitonin to C-reactive protein and leukocytosis for the detection of serious bacterial infections in children presenting with fever without source: a systematic review and meta-analysis.

PubMed

Yo, Chia-Hung; Hsieh, Pei-Shan; Lee, Si-Huei; Wu, Jiunn-Yih; Chang, Shy-Shin; Tasi, Kuang-Chau; Lee, Chien-Chang

2012-11-01

We determine the usefulness of the procalcitonin for early identification of young children at risk for severe bacterial infection among those presenting with fever without source. The design was a systematic review and meta-analysis of diagnostic studies. Data sources were searches of MEDLINE and EMBASE in April 2011. Included were diagnostic studies that evaluated the diagnostic value of procalcitonin alone or compared with other laboratory markers, such as C-reactive protein or leukocyte count, to detect severe bacterial infection in children with fever without source who were aged between 7 days and 36 months. Eight studies were included (1,883 patients) for procalcitonin analysis, 6 (1,265 patients) for C-reactive protein analysis, and 7 (1,649 patients) for leukocyte analysis. The markers differed in their ability to predict serious bacterial infection: procalcitonin (odds ratio [OR] 10.6; 95% confidence interval [CI] 6.9 to 16.0), C-reactive protein (OR 9.83; 95% CI 7.05 to 13.7), and leukocytosis (OR 4.26; 95% CI 3.22 to 5.63). The random-effect model was used for procalcitonin analysis because heterogeneity across studies existed. Overall sensitivity was 0.83 (95% CI 0.70 to 0.91) for procalcitonin, 0.74 (95% CI 0.65 to 0.82) for C-reactive protein, and 0.58 (95% CI 0.49 to 0.67) for leukocyte count. Overall specificity was 0.69 (95% CI 0.59 to 0.85) for procalcitonin, 0.76 (95% CI 0.70 to 0.81) for C-reactive protein, and 0.73 (95% CI 0.67 to 0.77) for leukocyte count. Procalcitonin performs better than leukocyte count and C-reactive protein for detecting serious bacterial infection among children with fever without source. Considering the poor pooled positive likelihood ratio and acceptable pooled negative likelihood ratio, procalcitonin is better for ruling out serious bacterial infection than for ruling it in. Existing studies do not define how best to combine procalcitonin with other clinical information. Copyright © 2012. Published by Mosby, Inc.

Characterization of Epiphytic Bacterial Communities from Grapes, Leaves, Bark and Soil of Grapevine Plants Grown, and Their Relations

PubMed Central

Martins, Guilherme; Lauga, Béatrice; Miot-Sertier, Cécile; Mercier, Anne; Lonvaud, Aline; Soulas, Marie-Louise; Soulas, Guy; Masneuf-Pomarède, Isabelle

2013-01-01

Despite its importance in plant health and crop quality, the diversity of epiphytic bacteria on grape berries and other plant parts, like leaves and bark, remains poorly described, as does the role of telluric bacteria in plant colonization. In this study, we compare the bacterial community size and structure in vineyard soils, as well as on grapevine bark, leaves and berries. Analyses of culturable bacteria revealed differences in the size and structure of the populations in each ecosystem. The highest bacteria population counts and the greatest diversity of genera were found in soil samples, followed by bark, grapes and leaves. The identification of isolates revealed that some genera – Pseudomonas, Curtobacterium, and Bacillus – were present in all ecosystems, but in different amounts, while others were ecosystem-specific. About 50% of the genera were common to soil and bark, but absent from leaves and grapes. The opposite was also observed: grape and leaf samples presented 50% of genera in common that were absent from trunk and soil. The bacterial community structure analyzed by T-RFLP indicated similarities between the profiles of leaves and grapes, on the one hand, and bark and soil, on the other, reflecting the number of shared T-RFs. The results suggest an interaction between telluric bacterial communities and the epiphytic bacteria present on the different grapevine parts. PMID:24023666

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

PubMed

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

2000-12-01

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

Air, water, and surface bacterial contamination in a university-hospital autopsy room.

PubMed

Maujean, Géraldine; Malicier, Daniel; Fanton, Laurent

2012-03-01

Today, little is known about the bacteriological environment of the autopsy room and its potential interest for medico-legal practices. Seven hundred fifty microbiological samples were taken from surface (n = 660), air (n = 48), and water (n = 42) to evaluate it in a French University Forensic Department. Median bacterial counts were compared before and during autopsy for air samples, and before and after autopsy for surface samples, using Wilcoxon matched pairs signed ranks test. Bacterial identification relied on traditional phenotypic methods. Bacterial counts in the air were low before autopsy, increased significantly during procedure, and seemed more linked to the number of people in the room than to an important production of aerosol-containing bacteria. Despite cleaning, human fecal flora was omnipresent on surfaces, which revealed insufficient disinfection. Bacteriological sampling is an easy way to monitor cleaning practices in postmortem rooms, but chiefly a way to improve the reliability of medico-legal proofs of infectious deaths. © 2012 American Academy of Forensic Sciences.

The impact of shrimp farming effluent on bacterial communities in mangrove waters, Ceará, Brazil.

PubMed

Sousa, O V; Macrae, A; Menezes, F G R; Gomes, N C M; Vieira, R H S F; Mendonça-Hagler, L C S

2006-12-01

The effects of shrimp farm effluents on bacterial communities in mangroves have been infrequently reported. Classic and molecular biology methods were used to survey bacterial communities from four mangroves systems. Water temperature, salinity, pH, total heterotrophic bacteria and maximum probable numbers of Vibrio spp. were investigated. Genetic profiles of bacterial communities were also characterized by polymerase chain reaction (PCR) amplification of eubacterial and Vibrio 16S rDNA using denaturing gradient gel electrophoresis (DGGE). Highest heterotrophic counts were registered in the mangrove not directly polluted by shrimp farming. The Enterobacteriaceae and Chryseomonas luteola dominated the heterotrophic isolates. Vibrio spp. pathogenic to humans and shrimps were identified. Eubacterial genetic profiles suggest a shared community structure independent of mangrove system. Vibrio genetic profiles were mangrove specific. Neither microbial counts nor genetic profiling revealed a significant decrease in species richness associated with shrimp farm effluent. The complex nature of mangrove ecosystems and their microbial communities is discussed.

Rapid classification of hairtail fish and pork freshness using an electronic nose based on the PCA method.

PubMed

Tian, Xiu-Ying; Cai, Qiang; Zhang, Yong-Ming

2012-01-01

We report a method for building a simple and reproducible electronic nose based on commercially available metal oxide sensors (MOS) to monitor the freshness of hairtail fish and pork stored at 15, 10, and 5 °C. After assembly in the laboratory, the proposed product was tested by a manufacturer. Sample delivery was based on the dynamic headspace method, and two features were extracted from the transient response of each sensor using an unsupervised principal component analysis (PCA) method. The compensation method and pattern recognition based on PCA are discussed in the current paper. PCA compensation can be used for all storage temperatures, however, pattern recognition differs according to storage conditions. Total volatile basic nitrogen (TVBN) and aerobic bacterial counts of the samples were measured simultaneously with the standard indicators of hairtail fish and pork freshness. The PCA models based on TVBN and aerobic bacterial counts were used to classify hairtail fish samples as "fresh" (TVBN ≤ 25 g and microbial counts ≤ 10(6) cfu/g) or "spoiled" (TVBN ≥ 25 g and microbial counts ≥ 10(6) cfu/g) and pork samples also as "fresh" (TVBN ≤ 15 g and microbial counts ≤ 10(6) cfu/g) or "spoiled" (TVBN ≥ 15 g and microbial counts ≥ 10(6) cfu/g). Good correlation coefficients between the responses of the electronic nose and the TVBN and aerobic bacterial counts of the samples were obtained. For hairtail fish, correlation coefficients were 0.97 and 0.91, and for pork, correlation coefficients were 0.81 and 0.88, respectively. Through laboratory simulation and field application, we were able to determine that the electronic nose could help ensure the shelf life of hairtail fish and pork, especially when an instrument is needed to take measurements rapidly. The results also showed that the electronic nose could analyze the process and level of spoilage for hairtail fish and pork.

Reduced Bacterial Colony Count of Anaerobic Bacteria Is Associated with a Worsening in Lung Clearance Index and Inflammation in Cystic Fibrosis

PubMed Central

Bradley, Judy M.; Johnston, Elinor; McGrath, Stephanie; McIlreavey, Leanne; Rowan, Stephen; Reid, Alastair; Bradbury, Ian; Einarsson, Gisli

2015-01-01

Anaerobic bacteria have been identified in abundance in the airways of cystic fibrosis (CF) subjects. The impact their presence and abundance has on lung function and inflammation is unclear. The aim of this study was to investigate the relationship between the colony count of aerobic and anaerobic bacteria, lung clearance index (LCI), spirometry and C-Reactive Protein (CRP) in patients with CF. Sputum and blood were collected from CF patients at a single cross-sectional visit when clinically stable. Community composition and bacterial colony counts were analysed using extended aerobic and anaerobic culture. Patients completed spirometry and a multiple breath washout (MBW) test to obtain LCI. An inverse correlation between colony count of aerobic bacteria (n = 41, r = -0.35; p = 0.02), anaerobic bacteria (n = 41, r = -0.44, p = 0.004) and LCI was observed. There was an inverse correlation between colony count of anaerobic bacteria and CRP (n = 25, r = -0.44, p = 0.03) only. The results of this study demonstrate that a lower colony count of aerobic and anaerobic bacteria correlated with a worse LCI. A lower colony count of anaerobic bacteria also correlated with higher CRP levels. These results indicate that lower abundance of aerobic and anaerobic bacteria may reflect microbiota disruption and disease progression in the CF lung. PMID:25992575

Reduced bacterial colony count of anaerobic bacteria is associated with a worsening in lung clearance index and inflammation in cystic fibrosis.

PubMed

O'Neill, Katherine; Bradley, Judy M; Johnston, Elinor; McGrath, Stephanie; McIlreavey, Leanne; Rowan, Stephen; Reid, Alastair; Bradbury, Ian; Einarsson, Gisli; Elborn, J Stuart; Tunney, Michael M

2015-01-01

Anaerobic bacteria have been identified in abundance in the airways of cystic fibrosis (CF) subjects. The impact their presence and abundance has on lung function and inflammation is unclear. The aim of this study was to investigate the relationship between the colony count of aerobic and anaerobic bacteria, lung clearance index (LCI), spirometry and C-Reactive Protein (CRP) in patients with CF. Sputum and blood were collected from CF patients at a single cross-sectional visit when clinically stable. Community composition and bacterial colony counts were analysed using extended aerobic and anaerobic culture. Patients completed spirometry and a multiple breath washout (MBW) test to obtain LCI. An inverse correlation between colony count of aerobic bacteria (n = 41, r = -0.35; p = 0.02), anaerobic bacteria (n = 41, r = -0.44, p = 0.004) and LCI was observed. There was an inverse correlation between colony count of anaerobic bacteria and CRP (n = 25, r = -0.44, p = 0.03) only. The results of this study demonstrate that a lower colony count of aerobic and anaerobic bacteria correlated with a worse LCI. A lower colony count of anaerobic bacteria also correlated with higher CRP levels. These results indicate that lower abundance of aerobic and anaerobic bacteria may reflect microbiota disruption and disease progression in the CF lung.

Histological observations in the Hawaiian reef coral, Porites compressa, affected by Porites bleaching with tissue loss

USGS Publications Warehouse

Sudek, M.; Work, Thierry M.; Aeby, G.S.; Davy, S.K.

2012-01-01

The scleractinian finger coral Porites compressa is affected by the coral disease Porites bleaching with tissue loss (PBTL). This disease initially manifests as bleaching of the coenenchyme (tissue between polyps) while the polyps remain brown with eventual tissue loss and subsequent algal overgrowth of the bare skeleton. Histopathological investigation showed a loss of symbiont and melanin-containing granular cells which was more pronounced in the coenenchyme than the polyps. Cell counts confirmed a 65% reduction in symbiont density. Tissue loss was due to tissue fragmentation and necrosis in affected areas. In addition, a reduction in putative bacterial aggregate densities was found in diseased samples but no potential pathogens were observed.

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

Microbiological changes associated with dental prophylaxis.

PubMed

Goodson, J Max; Palys, Michael D; Carpino, Elizabeth; Regan, Elizabeth O; Sweeney, Michael; Socransky, Sigmund S

2004-11-01

Despite the common application of dental prophylaxis as part of patient therapy, there is little reported that describes the microbiological impact of this treatment. The authors gave 20 healthy college-aged subjects three dental prophylaxes with a fluoride-containing prophylaxis paste during a two-week period and instructed them in oral hygiene. They evaluated the microbiological composition of dental plaque samples collected before and after treatment using DNA probe analysis. They analyzed 40 representative bacterial species in seven bacterial complexes by checkerboard DNA-DNA hybridization assay techniques. After three dental prophylaxes, the patients' mean Gingival Index score decreased from 0.82 to 0.77, the mean Plaque Index score decreased from 0.72 to zero, and the total number of bacteria per tooth decreased to approximately one-third of the original number. The authors computed two different measures of bacterial presence. The reduction in bacterial numbers was statistically significant and occurred in many species. Bacterial proportion (DNA percentage or percentage of the bacteria per tooth) did not change significantly. Greater reductions in bacterial count occurred in species that showed high numbers before treatment. The total bacterial count decreased by approximately 72 percent of its original level before prophylaxis was initiated. Professional dental prophylaxis did not target any particular bacteria or bacterial groups but removed bacteria nonspecifically and in proportion to their initial numbers. Repeated dental prophylaxes effect a reduction in bacterial amount that is commensurate with the initial amount, but they do does not alter composition. This suggests that mild gingivitis may be a bacterially nonspecific effect of plaque accumulation and emphasizes the need for regular plaque removal to maintain optimal gingival health.

Microbiology of processed edible insect products - Results of a preliminary survey.

PubMed

Grabowski, Nils Th; Klein, Günter

2017-02-21

Little is known of the microbiology of processed insect products. The present survey analysed a total of n=38 samples of deep-fried and spiced (Acheta domesticus, Locusta migratoria, and Omphisa fuscidentalis), cooked in soy sauce ("tsukudani"; Oxya yezoensis, Vespula flaviceps, and Bombyx mori), dried (A. domesticus, L. migatoria, Alphitobius diaperinus, Tenebrio molitor, B. mori, Hermetia illucens, and Musca domestica), powdered (H. illucens, T. molitor) and other (incl. deep-frozen B. mori and honeybee pollen) insect products microbiologically (total bacterial count [TBC], Enterobacteriaceae, staphylococci, bacilli, and yeasts and moulds counts, salmonellae, Listeria monocytogenes, and Escherichia coli). Although each product type revealed a microbiological profile of its own, dried and powdered insects ("class I") displayed markedly higher counts than the deep-fried and cooked ones ("class II"). Thresholds between class I and II products were estimated at 4.0 (TBC), 1.0 (Enterobacteriaceae, yeasts and moulds), 2.5 (staphylococci), and 3.0lgcfu/g (bacilli). All samples were negative for salmonellae, L. monocytogenes, E. coli and Stapyhlococcus aureus, but dried and powdered insects, as well as pollen, contained B. cereus, coliforms, Serratia liquefaciens, Listeria ivanovii, Mucor spp., Aspergillus spp., Penicillium spp., and Cryptococcus neoformans. Comparing the results with the hygiene criteria for edible insects proposed by Belgium and the Netherlands, class I products failed to comply with many bacterial count limits despite the absence of classical food pathogens. Therefore, class I products should always be consumed after another heating step as indicated by the manufacturer, until drying techniques are able to ensure lower bacterial counts. Copyright © 2016 Elsevier B.V. All rights reserved.

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.

Bacterial flora of the sigmoid neovagina.

PubMed Central

Toolenaar, T A; Freundt, I; Wagenvoort, J H; Huikeshoven, F J; Vogel, M; Jeekel, H; Drogendijk, A C

1993-01-01

The bacterial microbiota of 15 sigmoid neovaginas, created in patients with congenital vaginal aplasia or male transsexualism, was studied. No specimen was sterile, and only normal inhabitants of the colon were cultured. The total counts of bacteria were lower than those reported for healthy sigmoid colons. PMID:8308126

Real-time measurements of airborne biologic particles using fluorescent particle counter to evaluate microbial contamination: results of a comparative study in an operating theater.

PubMed

Dai, Chunyang; Zhang, Yan; Ma, Xiaoling; Yin, Meiling; Zheng, Haiyang; Gu, Xuejun; Xie, Shaoqing; Jia, Hengmin; Zhang, Liang; Zhang, Weijun

2015-01-01

Airborne bacterial contamination poses a risk for surgical site infection, and routine surveillance of airborne bacteria is important. Traditional methods for detecting airborne bacteria are time consuming and strenuous. Measurement of biologic particle concentrations using a fluorescent particle counter is a novel method for evaluating air quality. The current study was to determine whether the number of biologic particles detected by the fluorescent particle counter can be used to indicate airborne bacterial counts in operating rooms. The study was performed in an operating theater at a university hospital in Hefei, China. The number of airborne biologic particles every minute was quantified using a fluorescent particle counter. Microbiologic air sampling was performed every 30 minutes using an Andersen air sampler (Pusong Electronic Instruments, Changzhou, China). Correlations between the 2 different methods were analyzed by Pearson correlation coefficients. A significant correlation was observed between biologic particle and bacterial counts (Pearson correlation coefficient = 0.76), and the counting results from 2 methods both increased substantially between operations, corresponding with human movements in the operating room. Fluorescent particle counters show potential as important tools for monitoring bacterial contamination in operating theatres. Copyright © 2015 Association for Professionals in Infection Control and Epidemiology, Inc. Published by Elsevier Inc. All rights reserved.

Effect of Manuka honey gel on the transforming growth factor β1 and β3 concentrations, bacterial counts and histomorphology of contaminated full-thickness skin wounds in equine distal limbs.

PubMed

Bischofberger, A S; Dart, C M; Horadagoda, N; Perkins, N R; Jeffcott, L B; Little, C B; Dart, A J

2016-01-01

To investigate the effect of 66% Manuka honey gel on the concentrations of transforming growth factor (TGF)-β1 and TGF-β3, bacterial counts and histomorphology during healing of contaminated equine distal limb wounds. In this experimental study of 10 Standardbred horses, five full-thickness skin wounds (2 × 1.5 cm) were created on one metacarpus and six similar wounds were created on the contralateral metacarpus. Wounds were assigned to three groups: non-contaminated control wounds; contaminated control wounds; contaminated wounds treated daily with 1 mL Manuka honey gel topically for 10 days. For the contaminated wounds, faeces were applied for 24 h after wound creation. In five horses wounds were bandaged and in the other five horses wounds were left without a bandage. Biopsies were taken on days 1, 2, 7 and 10 after wounding to evaluate the effects of Manuka honey gel, wound contamination and bandaging on TGF-β1 and TGF-β3 concentrations, aerobic and anaerobic bacterial counts, and histomorphology. Manuka honey gel had no significant effect on TGF-β1 and TGF-β3 concentrations or wound bacterial counts. Manuka honey gel decreased wound inflammation (days 7, 10), increased angiogenesis (days 2, 7, 10), increased fibrosis and collagen organisation (day 7) and increased epithelial hyperplasia (days 7, 10). Treatment with Manuka honey gel resulted in a more organised granulation tissue bed early in wound repair, which may contribute to enhanced healing of equine distal limb wounds. © 2016 Australian Veterinary Association.

A Prospective Multicenter Study of Leukopenia in Infants Younger Than Ninety Days With Fever Without Source.

PubMed

Gomez, Borja; Mintegi, Santiago; Benito, Javier

2016-01-01

Little is known about the value of leukopenia for assessing the risk of having a bacterial infection in young febrile infants. Infants younger than 90 days with fever without source were prospectively recruited between October 2011 and September 2013 in 19 Spanish Pediatric Emergency Departments. We analyzed the prevalence of invasive bacterial infection (IBI, positive blood or cerebrospinal fluid culture) and non-IBI (urinary tract infections and any other microbiologically confirmed bacterial infection excluding IBIs) by leukocyte count and general appearance. Among the 3401 infants recruited, 680 were diagnosed with non-IBIs (19.9%) and 107 with IBIs (3.1%). Overall, 244 infants had leukopenia (<5000 cells/mcL), 2369 a normal leukocyte count and 790 leukocytosis (>15,000 cells/mcL). Among the 3034 well-appearing patients, those with leukopenia had a lower prevalence of non-IBI [8.1% vs. 14.7%; odds ratio (OR) 0.51 (95% confidence interval (CI): 0.29-0.88)] and a similar prevalence of IBI [2.5% vs. 2.0%; OR, 1.20 (95% CI: 0.44-3.44)] compared with those with a normal leukocyte count. Among the 367 not-well-appearing infants, those with leukopenia had a similar prevalence of non-IBI [8.9% vs. 14.7%; OR, 0.57 (95% CI: 0.16-1.79)] and a higher prevalence of IBI [17.8% vs. 6.9%; OR, 2.90 (95% CI: 1.06-7.78)]. In the subgroup of well-appearing infants 22-90 days old without leukocyturia according to urine dipstick results, prevalence of both non-IBIs and IBIs was similar in patients with leukopenia and those with a normal leukocyte count. Leukopenia in well-appearing young febrile infants should not be considered a risk factor for having a bacterial infection.

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.

Age-specific application of neutrophil-to-lymphocyte ratio in meningitis: a nationwide study.

PubMed

Mentis, A-F A; Kyprianou, M A; Tzanakaki, G

2017-09-01

Cerebrospinal fluid (CSF) neutrophil counts and neutrophil-to-lymphocyte ratio (NLR) are useful in distinguishing bacterial and viral meningitis. Given that meningitis is clinically heterogeneous with regard to age, here we investigated the validity of the CSF NLR and neutrophil assay according to age group. Data from the nationwide referral of >4,000 meningitis cases to the Hellenic Meningitis Reference Laboratory between 2006 and 2013 were examined. CSF NLR and neutrophil counts were stratified according to age, and assay performance was determined using previous cut-off values of 2 and 287 cells/μl for CSF NLR and neutrophils respectively. The distribution of bacterial versus viral meningitis was heterogenous across age groups, with a low proportion of bacterial meningitis in patients aged 5-14. CSF neutrophil count and NLR were significantly more discriminatory for bacterial meningitis in patients aged over 14 years than those aged 0-14. The odds ratio (OR), sensitivity, specificity and positive predictive value (PPV) were significantly higher in older patients for both biomarkers. When combined, the false-positive and false-negative detection of bacterial meningitis was 3.9 and 8.5% respectively, and the OR of 262.2 was 2.5-fold greater than expected from a multiplicative effect alone in patients aged >14 years. Care is required when applying diagnostic tests for meningitis in different age groups because of patient heterogeneity. This is the first description of the age distribution of meningitis cases in Greece, and knowledge of the age-related distribution of neutrophils and NLR in meningitis cases could help towards developing age-specific meningitis diagnostic assays.

Antibacterial Activity of Diode Laser and Sodium Hypochlorite in Enterococcus Faecalis-Contaminated Root Canals.

PubMed

Sohrabi, Khosrow; Sooratgar, Aidin; Zolfagharnasab, Kaveh; Kharazifard, Mohammad Javad; Afkhami, Farzaneh

2016-01-01

The aim of the present in vitro study was to evaluate the disinfection ability of 980-nm diode laser in comparison with sodium hypochlorite (NaOCl) as a common root canal irrigant in canals infected with Enterococcus faecalis (E. faecalis). The root canals of 18 extracted single-rooted premolars were prepared by rotary system. After decoronation, the roots were autoclaved. One specimen was chosen for the negative control, and the remaining teeth were incubated with E. faecalis suspension for two weeks. Subsequently, one specimen was selected as the positive control and the remaining samples were divided into two groups (n=8). The samples of the first group were irrigated with 5.25% NaOCl and the second group were treated with a 980-nm diode laser. Microbial samples were taken from the root canals and bacterial cultivation was carried out. The average value and the standard deviation of colony-forming units (CFU) of each specimen were measured using descriptive statistics. The student's t-test was used to compare the reduction in CFU in each group. The equality of variance of CFU was measured by the Levene's test. NaOCl resulted in 99.87% removal of the bacteria and showed significantly more antibacterial effect compared to the 980-nm diode laser which led to 96.56% bacterial reduction (P<0.05). Although 5.25% NaOCl seems to reduce E. faecalis more effectively, the diode laser also reduced the bacterial count. Therefore a 980-nm diode laser could be considered as a complementary disinfection method in root canal treatment.

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/).

Chemomechanical preparation by hand instrumentation and by Mtwo engine-driven rotary files, an ex vivo study.

PubMed

Krajczár, Károly; Tigyi, Zoltán; Papp, Viktória; Marada, Gyula; Sára, Jeges; Tóth, Vilmos

2012-07-01

To compare the disinfecting efficacy of the sodium hypochlorite irrigation by root canal preparation with stainless steel hand files, taper 0.02 and nickel-titanium Mtwo files with taper 0.04-0.06. 40 extracted human teeth were sterilized, and then inoculated with Enterococcus faecalis (ATCC 29212). After 6 day incubation time the root canals were prepared by hand with K-files (n=20) and by engine-driven Mtwo files (VDW, Munich, Germany) (n=20). Irrigation was carried out with 2.5% NaOCl in both cases. Samples were taken and determined in colony forming units (CFU) from the root canals before and after the preparation with instruments #25 and #35. Significant reduction in bacterial count was determined after filing at both groups. The number of bacteria kept on decreasing with the extension of apical preparation diameter. There was no significant difference between the preparation sizes in the bacterial counts after hand or engine-driven instrumentation at the same apical size. Statistical analysis was carried out with Mann-Whitney test, paired t-test and independent sample t-test. Significant reduction in CFU was achieved after the root canal preparation completed with 2.5% NaOCl irrigation, both with stainless steel hand or nickel-titanium rotary files. The root canal remained slightly infected after chemo mechanical preparation in both groups. Key words:Chemomechanical preparation, root canal disinfection, nickel-titanium, conicity, greater taper, apical size.

Chemomechanical preparation by hand instrumentation and by Mtwo engine-driven rotary files, an ex vivo study

PubMed Central

Krajczár, Károly; Tigyi, Zoltán; Papp, Viktória; Sára, Jeges; Tóth, Vilmos

2012-01-01

Objective: To compare the disinfecting efficacy of the sodium hypochlorite irrigation by root canal preparation with stainless steel hand files, taper 0.02 and nickel-titanium Mtwo files with taper 0.04-0.06. Study Design: 40 extracted human teeth were sterilized, and then inoculated with Enterococcus faecalis (ATCC 29212). After 6 day incubation time the root canals were prepared by hand with K-files (n=20) and by engine-driven Mtwo files (VDW, Munich, Germany) (n=20). Irrigation was carried out with 2.5% NaOCl in both cases. Samples were taken and determined in colony forming units (CFU) from the root canals before and after the preparation with instruments #25 and #35. Results: Significant reduction in bacterial count was determined after filing at both groups. The number of bacteria kept on decreasing with the extension of apical preparation diameter. There was no significant difference between the preparation sizes in the bacterial counts after hand or engine-driven instrumentation at the same apical size. Statistical analysis was carried out with Mann-Whitney test, paired t-test and independent sample t-test. Conclusions: Significant reduction in CFU was achieved after the root canal preparation completed with 2.5% NaOCl irrigation, both with stainless steel hand or nickel-titanium rotary files. The root canal remained slightly infected after chemo mechanical preparation in both groups. Key words:Chemomechanical preparation, root canal disinfection, nickel-titanium, conicity, greater taper, apical size. PMID:24558545

Survival and enumeration of the fecal indicators Bifidobacterium adolescentis and Escherichia coli in a tropical rain forest watershed.

PubMed Central

Carrillo, M; Estrada, E; Hazen, T C

1985-01-01

The density of Bifidobacterium spp., fecal coliforms, Escherichia coli, and total anaerobic bacteria, acridine orange direct counts, percentages of total bacterial community activity and respiration, and 12 physical and chemical parameters were measured simultaneously at six sites for 12 months in the Mameyes River rain forest watershed, Puerto Rico. The densities of all bacteria were higher than those reported for uncontaminated temperate rivers, even though other water quality parameters would indicate that all uncontaminated sites were oligotrophic. The highest densities for all indicator bacteria were at the site receiving sewage effluent; however, the highest elevation site in the watershed had the next highest densities. Correlations between bacterial densities, nitrates, temperature, phosphates, and total phosphorus indicated that all viable counts were related to nutrient levels, regardless of the site sampled. In situ diffusion chamber studies at two different sites indicated that E. coli could survive, remain physiologically active, and regrow at rates that were dependent on nutrient levels of the ambient waters. Bifidobacterium adolescentis did not survive at either site but did show different rates of decline and physiological activity at the two sites. Bifidobacteria show promise as a better indicator of recent fecal contamination in tropical freshwaters than E. coli or fecal coliforms; however, the YN-6 medium did not prove to be effective for enumeration of bifidobacteria. The coliform maximum contaminant levels for assessing water usability for drinking and recreation appear to be unworkable in tropical freshwaters. PMID:3901921

Bacterial and fungal microbiota of spontaneously fermented Chinese products, Rubing milk cake and Yan-cai vegetable pickles.

PubMed

Liu, Xin; Kuda, Takashi; Takahashi, Hajime; Kimura, Bon

2018-06-01

The Rubing milk cake from Yunnan and the Yan-cai vegetable pickles from Guangdong are traditional spontaneously fermented foods in China. We evaluated the microbial properties of these products with the analysis of their bacterial and fungal microbiota using classical culture-dependent and culture-independent methods, including a 16S rDNA gene (V4) and an internal transcribed spacer (ITS) region pyrosequencing method with MiSeq system. The viable lactic acid bacteria (LAB) count was 8 and 6 log colony-forming units (CFU)/g in Rubing and Yan-cai samples, respectively. The yeast count was approximately 100-1000 times less than the LAB count in most samples, except one Yan-cai sample. In addition, the gram-negative rod count in half of the samples was similar to the LAB count. Pyrosequencing results revealed the high abundance (10%-20%) of gram-negative Pseudomonas spp. and Enterobacteriaceae in these samples. These results suggest that some of these traditional foods are undesirable as ready-to-eat (RTE) foods, even when these are typical lactic acid fermented foods. Copyright © 2017 Elsevier Ltd. All rights reserved.

Examination of body fluids.

PubMed

Feldman, B F; Ruehl, W W

1984-04-01

In dogs, the pericardial sac contains about 0.3 ml, and the pleural and peritoneal cavities 0-15 ml of clear, straw-colored fluid of pH 7.4, specific gravity 1.016, protein content less than 3.0 g/dl and cell count less than 3000/microliter. Fat can be cleared from chylous fluid with NaOH and ether. Inflammation is indicated by a cell count greater than 3000/microliter. Amylase levels in peritoneal fluid are elevated in necrotizing pancreatitis. The percentage of polymorphonuclear WBC exceeds 50% in bacterial inflammations. Normal joints contain less than 1 ml highly viscid, clear or straw-colored synovial fluid with less than 1000 nucleated cells/microliter. Synovial fluid becomes flocculent and less viscid in septic and occasionally in immune-mediated arthritis, often with cell counts greater than 75,000/microliter, with 75-90% polymorphonuclear WBC. Cerebrospinal fluid is normally acellular, clear and colorless but may be red, yellow or brown with intracranial hematomas. Viral or aseptic meningitis is characterized by mononuclear cell counts of less than 500/microliter. In acute bacterial meningitis, nucleated cell counts are greater than 1000/microliter, with most being polymorphonuclear WBC. Gram staining of cerebrospinal fluid is not useful.

Surface topography of composite restorative materials following ultrasonic scaling and its Impact on bacterial plaque accumulation. An in-vitro SEM study.

PubMed

Hossam, A Eid; Rafi, A Togoo; Ahmed, A Saleh; Sumanth, Phani Cr

2013-06-01

This is an in vitro study to investigate the effects of ultrasonic scaling on the surface roughness and quantitative bacterial count on four different types of commonly used composite restorative materials for class V cavities. Nanofilled, hybrid, silorane and flowable composites were tested. Forty extracted teeth served as specimen and were divided into 4 groups of 10 specimens, with each group receiving a different treatment and were examined by a Field emission scanning electron microscope. Bacterial suspension was then added to the pellicle-coated specimens, and then bacterial adhesion was analyzed by using image analyzing program. Flowable and silorane-based composites showed considerably smoother surfaces and lesser bacterial count in comparison to other types, proving that bacterial adhesion is directly proportional to surface roughness. The use of ultrasonic scalers affects the surfaces of composite restorative materials. Routine periodontal scaling should be carried out very carefully, and polishing of the scaled surfaces may overcome the alterations in roughness, thus preventing secondary caries, surface staining, plaque accumulation and subsequent periodontal inflammation. How to cite this article: Eid H A, Togoo R A, Saleh A A, Sumanth C R. Surface Topography of Composite Restorative Materials following Ultrasonic Scaling and its Impact on Bacterial Plaque Accumulation. An In-Vitro SEM Study. J Int Oral Health 2013; 5(3):13-19.

Surface topography of composite restorative materials following ultrasonic scaling and its Impact on bacterial plaque accumulation. An in-vitro SEM study

PubMed Central

Hossam, A. Eid; Rafi, A. Togoo; Ahmed, A Saleh; Sumanth, Phani CR

2013-01-01

Background: This is an in vitro study to investigate the effects of ultrasonic scaling on the surface roughness and quantitative bacterial count on four different types of commonly used composite restorative materials for class V cavities. Materials & Methods: Nanofilled, hybrid, silorane and flowable composites were tested. Forty extracted teeth served as specimen and were divided into 4 groups of 10 specimens, with each group receiving a different treatment and were examined by a Field emission scanning electron microscope. Bacterial suspension was then added to the pellicle-coated specimens, and then bacterial adhesion was analyzed by using image analyzing program. Results: Flowable and silorane-based composites showed considerably smoother surfaces and lesser bacterial count in comparison to other types, proving that bacterial adhesion is directly proportional to surface roughness. Conclusion: The use of ultrasonic scalers affects the surfaces of composite restorative materials. Routine periodontal scaling should be carried out very carefully, and polishing of the scaled surfaces may overcome the alterations in roughness, thus preventing secondary caries, surface staining, plaque accumulation and subsequent periodontal inflammation. How to cite this article: Eid H A, Togoo R A, Saleh A A, Sumanth C R. Surface Topography of Composite Restorative Materials following Ultrasonic Scaling and its Impact on Bacterial Plaque Accumulation. An In-Vitro SEM Study. J Int Oral Health 2013; 5(3):13-19. PMID:24155597

Role of Clinical Presentations and Routine CSF Analysis in the Rapid Diagnosis of Acute Bacterial Meningitis in Cases of Negative Gram Stained Smears

PubMed Central

Fouad, Rabab; El-Kholy, Badawy; Yosry, Ayman

2014-01-01

Background and Aim. Bacterial meningitis is a lethal, disabling endemic disease needing prompt antibiotic management. Gram stained smears is rapid accurate method for diagnosis of bacterial meningitis. In cases of negative gram stained smears diagnosis is delayed till culture results. We aim to assess the role of clinical presentations and routine CSF analysis in the cost-effective rapid diagnosis of negative gram stained smears bacterial meningitis. Methods. Cross sectional study including 623 acute meningitis patients divided into two groups: bacterial meningitis and nonbacterial meningitis groups. The clinical presentations, systemic inflammatory parameters, and CSF analysis were evaluated and compared in both groups. Results. Altered conscious level, localizing neurological signs, Kernig's and Brudzinski's signs together with peripheral leucocytosis (>10.000/mm3), high CRP (>6) together with high CSF protein (>50 gl/dL), CSF neutrophilic count (≥50% of total CSF leucocytic count), and low CSF glucose level (<45 gm/dL) and CSF/serum glucose ≤0.6 were significantly diagnostic in bacterial meningitis patients. From the significant CSF analysis variables CSF protein carried the higher accuracy of diagnosis 78% with sensitivity 88% and specificity 72%. Conclusions. High CSF protein (>50 mg/dL) together with plasma inflammatory markers and CSF cytochemical parameters can diagnose bacterial meningitis in gram stain negative smear till culture results. PMID:24803939

Role of neutrophil to lymphocyte and monocyte to lymphocyte ratios in the diagnosis of bacterial infection in patients with fever.

PubMed

Naess, Are; Nilssen, Siri Saervold; Mo, Reidun; Eide, Geir Egil; Sjursen, Haakon

2017-06-01

To study the role of the neutrophil:lymphocyte ratio (NLR) and monocyte:lymphocyte ratio (MLR) in discriminating between different patient groups hospitalized for fever due to infection and those without infection. For 299 patients admitted to hospital for fever with unknown cause, a number of characteristics including NLR and MLR were recorded. These characteristics were used in a multiple multinomial regression analysis to estimate the probability of a final diagnostic group of bacterial, viral, clinically confirmed, or no infection. Both NLR and MLR significantly predicted final diagnostic group. Being highly correlated, however, both variables could not be retained in the same model. Both variables also interacted significantly with duration of fever. Generally, higher values of NLR and MLR indicated larger probabilities for bacterial infection and low probabilities for viral infection. Patients with septicemia had significantly higher NLR compared to patients with other bacterial infections with fever for less than one week. White blood cell counts, neutrophil counts, and C-reactive proteins did not differ significantly between septicemia and the other bacterial infection groups. NLR is a more useful diagnostic tool to identify patients with septicemia than other more commonly used diagnostic blood tests. NLR and MLR may be useful in the diagnosis of bacterial infection among patients hospitalized for fever.

Comparison of bacteria populations in clean and recycled sand used for bedding in dairy facilities.

PubMed

Kristula, M A; Rogers, W; Hogan, J S; Sabo, M

2005-12-01

Bedding samples were collected twice from commercial dairy free-stall facilities that used recycled sand and clean sand in both the summer and winter. Collection began on the day sand was taken from the pile (d 0) and placed in the free stalls, and continued for 5 to 7 additional days. The number of colonies per gram of bedding of gram-negative bacteria, coliforms, Streptococcus spp., and Klebsiella spp. were estimated for each sand sample as well as amounts of dry and organic matter. Clean sand (CS) and recycled sand (RS) had the same bacterial counts when compared at any sampling time. The mean counts of bacterial populations did vary over the course of the study in both CS and RS. There was a significant increase in bacterial counts from d 0 to d 1 for gram-negative bacteria, coliforms, and Streptococcus spp. in both winter and summer. Counts of gram-negative bacteria, coliforms, Klebsiella spp., and Streptococcus spp. did not differ from d 1 to 7 in the winter. Total counts of gram-negative bacteria did not differ from d 1 to 7 in the summer. On d 1 in the summer, coliform counts were lower than at d 5 to 7, and Klebsiella spp. counts were lower than on d 3 to 7. Streptococcus spp. counts were high on d 1 and were constant through d 7 in both winter and summer trials. The number of coliform and Klebsiella spp. in both CS and RS was below the threshold thought to cause mastitis during the sampling times. The number of Streptococcus spp. was high in both CS and RS during the sampling periods. Other management factors need to be identified to decrease the number of Streptococcus spp. in bedding. Recycled sand had a higher organic matter and lower dry matter compared with CS in winter and summer. The results for this study were obtained from multiple herd comparisons, and herd was a significant effect suggesting that different management systems influence the number and types of bacteria in both CS and RS.

Evaluation of a novel gel-based ureteral stent with biofilm-resistant characteristics.

PubMed

Rosman, Brian M; Barbosa, Joao A B A; Passerotti, Carlo P; Cendron, Marc; Nguyen, Hiep T

2014-06-01

Current ureteral stents, while effective at maintaining a ureteral lumen, provide a substrate for bacterial growth. This propensity for biofilm formation may be a nidus for bacterial growth leading to infection and a reason for early removal of a stent before it is clinically indicated. A newly devised stent, composed of a highly hydrated, partially hydrolyzed polyacrylonitrile polymer, is believed to have bacterial resistant properties. The objective of this study is to evaluate the biofilm growth and bacterial resistant properties of this novel stent. Multiple 1 cm sections of the pAguaMedicina™ Pediatric Ureteral Stent (pAMS) (Q Urological, Natick, MA) and the conventional polymer stent (SS) (Boston Scientific, Natick, MA) were incubated for 3 days in the 3 different growth media. Afterward, J96 human pathogenic Escherichia coli was added. At 3, 6, 9, 12, and 15 days following bacterial inoculation, the stent segments were washed, sonicated, and analyzed for bacterial growth. Scanning electron microscopy (SEM) imaging was performed to assess biofilm formation. pAMS demonstrated significant reductions (43-71 %) in bacterial counts when compared to standard stents in all conditions tested. SEM imaging demonstrated biofilm formation on both types of stents in all media, with a relative reduction in apparent cell debris and bacteria on the pAMS. In this study, the gel-based stent shows a demonstrable reduction in bacterial counts and biofilm formation. The use of the pAMS may reduce the risk of infection associated with stent usage.

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…

Post-immunization leucocytosis and its implications for the management of febrile infants.

PubMed

Prentice, Sarah; Kamushaaga, Zephyrian; Nash, Stephen B; Elliott, Alison M; Dockrell, Hazel M; Cose, Stephen

2018-05-11

Clinical guidelines for management of infants with fever but no evident focus of infection recommend that those aged 1-3 months with a white cell count >15 × 10 9 /l have a full septic screen and be admitted for parenteral antibiotics. However, there is limited information about leucocyte changes following routine immunization, a common cause of fever. We investigated white cell counts shortly after routine immunization in Ugandan infants under 3 months of age. White cell counts were measured in 212 healthy infants following routine immunizations (DTwP-HepB-Hib, oral polio and pneumococcal conjugate 7 vaccines) received prior to 3 months of age. Mean leucocyte counts increased from 9.03 × 10 9 /l (95% confidence interval 8.59-9.47 × 10 9 /l) pre-immunizations to 16.46 × 10 9 /l (15.4-17.52 × 10 9 /l) at one-day post-immunizations at 6 weeks of age, and 15.21 × 10 9 /l (14.07-16.36 × 10 9 /l) at one-day post-immunizations at 10 weeks of age. The leucocytosis was primarily a neutrophilia, with neutrophil percentages one-day post-immunization of 49% at 6 weeks of age and 46% at 10 weeks of age. White cell parameters returned to baseline by two-days post-immunization. No participant received antibiotics when presenting with isolated fever post-immunization and all remained well at follow-up. In our study almost half the children <3 months old presenting with fever but no evident focus of infection at one-day post-immunization met commonly used criteria for full septic screen and admission for parenteral antibiotics, despite having no serious bacterial infection. These findings add to the growing body of literature that questions the utility of white blood cell measurement in identification of young infants at risk of serious bacterial infections, particularly in the context of recent immunizations, and suggest that further exploration of the effect of different immunization regimes on white cell counts is needed. This observational work was nested within a clinical trial, registration number ISRCTN59683017. Copyright © 2018 The Authors. Published by Elsevier Ltd.. All rights reserved.

Black soldier fly (Diptera: Stratiomyidae) larvae reduce Escherichia coli in dairy manure.

PubMed

Liu, Qiaolin; Tomberlin, Jeffery K; Brady, Jeff A; Sanford, Michelle R; Yu, Ziniu

2008-12-01

Escherichia coli labeled with a green fluorescent protein was inoculated into sterile dairy manure at 7.0 log cfu/g. Approximately 125 black soldier fly larvae were placed in manure inoculated and homogenized with E. coli. Manure inoculated with E. coli but without black soldier fly larvae served as the control. For the first experiment, larvae were introduced into 50, 75, 100, or 125 g sterilized dairy manure inoculated and homogenized with E. coli and stored 72 h at 27 degrees C. Black soldier fly larvae significantly reduced E. coli counts in all treatments. However, varying the amount of manure provided the black soldier fly larvae significantly affected their weight gain and their ability to reduce E. coli populations present. For the second experiment, larvae were introduced into 50 g manure inoculated with E. coli and stored for 72 h at 23, 27, 31, or 35 degrees C. Minimal bacterial growth was recorded in the control held at 35 degrees C and was excluded from the analysis. Black soldier fly larvae significantly reduced E. coli counts in manure held at remaining temperatures. Accordingly, temperature significantly influenced the ability of black soldier fly larvae to develop and reduce E. coli counts with greatest suppression occurring at 27 degrees C.

Population dynamics of two antilisterial cheese surface consortia revealed by temporal temperature gradient gel electrophoresis

PubMed Central

2010-01-01

Background Surface contamination of smear cheese by Listeria spp. is of major concern for the industry. Complex smear ecosystems have been shown to harbor antilisterial potential but the microorganisms and mechanisms involved in the inhibition mostly remain unclear, and are likely related to complex interactions than to production of single antimicrobial compounds. Bacterial biodiversity and population dynamics of complex smear ecosystems exhibiting antilisterial properties in situ were investigated by Temporal temperature gradient gel electrophoresis (TTGE), a culture independent technique, for two microbial consortia isolated from commercial Raclette type cheeses inoculated with defined commercial ripening cultures (F) or produced with an old-young smearing process (M). Results TTGE revealed nine bacterial species common to both F and M consortia, but consortium F exhibited a higher diversity than consortium M, with thirteen and ten species, respectively. Population dynamics were studied after application of the consortia on fresh-produced Raclette cheeses. TTGE analyses revealed a similar sequential development of the nine species common to both consortia. Beside common cheese surface bacteria (Staphylococcus equorum, Corynebacterium spp., Brevibacterium linens, Microbacterium gubbeenense, Agrococcus casei), the two consortia contained marine lactic acid bacteria (Alkalibacterium kapii, Marinilactibacillus psychrotolerans) that developed early in ripening (day 14 to 20), shortly after the growth of staphylococci (day 7). A decrease of Listeria counts was observed on cheese surface inoculated at day 7 with 0.1-1 × 102 CFU cm-2, when cheeses were smeared with consortium F or M. Listeria counts went below the detection limit of the method between day 14 and 28 and no subsequent regrowth was detected over 60 to 80 ripening days. In contrast, Listeria grew to high counts (105 CFU cm-2) on cheeses smeared with a defined surface culture. Conclusions This work reports the first population dynamics study of complex smear ecosystems exhibiting in situ antilisterial activity. TTGE revealed the presence of marine lactic acid bacteria that are likely related to the strong Listeria inhibition, as their early development in the smear occurred simultaneously with a decrease in Listeria cell count. PMID:20222967

Urinary tract infections in women with urogynaecological symptoms.

PubMed

Lakeman, Marielle M E; Roovers, Jan-Paul W R

2016-02-01

Urinary tract infections are common in the field of urogynaecology. Women with persistent urinary symptoms seem more likely to have bacteriuria despite negative cultures. In this review, we will give an overview of the recent insights on the relationship between urinary tract infection and persistent urinary symptoms and possible new therapeutic options. Recently published articles evaluated the prevalence of low-count bacteriuria (≥10 CFU/ml) or intracellular bacterial communities in women with overactive bladder symptoms (OAB). Differences in urinary microbioma observed in women with and without OAB symptoms were evaluated. In the light of these findings, current screening strategies were discussed and alternative screening methods for bacteriuria developed. Low-count bacteriuria (≥10 CFU/ml) seems to be more prevalent in women with OAB. Also intracellular bacterial communities are more commonly detected in these women. The microbioma found in women with urinary symptoms appeared to differ from healthy controls. The current screening methods might be insufficient as they are targeted at detecting uropathogenic Escherichia coli, mostly using a detection threshold of at least 10 CFU/ml and failing to detect intracellular bacterial communities. Studies evaluating the efficacy of treating women with low-count bacteriuria are limited but promising.

Direct viable count as test for toxicity assessment: the effects of four metals on a Salmonella enteritidis strain.

PubMed

Scoglio, M E; Di Pietro, A; Anzalone, C; Calimeri, S; Lo Giudice, D; Trimarchi, G R

2000-01-01

The toxicity of synthetic sewage containing increasing concentrations of arsenic (.125, .25, .5, 1.0 mg L-1), cadmium (.02, .05, .1, .2 mg L-1), lead (.2, .5, 1.0, 2.0 mg L-1) and nickel (.5, 1.0, 2.0, 4.0 mg L-1) has been investigated by determining the total direct count (TDC) and the direct viable count (DVC) of Salmonella enteritidis by means of an immunofluorescence technique (IFA). This has been done in order to evaluate the possibility of using the IFA technique to estimate the toxicity of complex effluents. Arsenic, cadmium and nickel produced a concentration-dependent reduction in the number of viable bacterial cells. This was more clear when the viable bacterial cells were considered than when only the culturable part was used. Lead did not show a concentration-dependent and reproducible effect. At the highest concentrations allowed by the Italian wastewater regulations, lead, cadmium, arsenic and nickel reduced the viable/total bacterial cells ratio to 74.5%, 68.5%, 28.4% and 6.9%, respectively. The toxic effects of the metals were also tested using the standard Microtox assay.

Antimicrobial Efficacy of Contact Lens Care Solutions Against Neutrophil-Enhanced Bacterial Biofilms

PubMed Central

Hinojosa, Jorge A.; Patel, Naiya B.; Zhu, Meifang; Robertson, Danielle M.

2017-01-01

Purpose Neutrophil-derived extracellular debris has been shown to accelerate bacterial biofilm formation on hydrogel and silicone hydrogel contact lens surfaces compared to lenses inoculated with bacteria alone. The purpose of this study was to evaluate the disinfection efficacy of four standard commercial contact lens cleaning regimens against neutrophil-enhanced bacterial biofilms formed on silicone hydrogel contact lenses. Methods Four reference strains were used: Pseudomonas aeruginosa, Serratia marcescens, Stenotrophomonas maltophilia, and Staphylococcus aureus. Human neutrophils were isolated from peripheral blood by venipuncture. Unworn Lotrafilcon B lenses were incubated overnight in each respective strain with stimulated neutrophils. Contact lenses were then cleaned using one of four contact lens care solutions according to manufacturer instructions. Bacterial viability was assessed by colony counts and confocal microscopy. Volume of residual debris on lens surfaces after cleaning was quantified using IMARIS software. Results All four solutions tested showed effective antimicrobial activity against each bacterial strain; however, substantial amounts of nonviable bacteria and cellular debris remained on the lens surface despite concomitant digital cleaning. Conclusions Necrotic cellular debris that accumulates under the posterior lens surface during wear of an inoculated contact lens is not fully removed during routine cleaning and disinfection. Translational Relevance The accumulation of residual cellular debris on the contact lens surface may contribute to new colonization of the lens and represents a significant risk factor for a contact lens–related adverse event. Additional studies are needed to correlate these findings with risk for corneal infiltrative and/or infectious events in a standard animal model. PMID:28473944

Establishment of the first international repository for transfusion-relevant bacteria reference strains: ISBT working party transfusion-transmitted infectious diseases (WP-TTID), subgroup on bacteria.

PubMed

Störmer, M; Arroyo, A; Brachert, J; Carrero, H; Devine, D; Epstein, J S; Gabriel, C; Gelber, C; Goodrich, R; Hanschmann, K-M; Heath, D G; Jacobs, M R; Keil, S; de Korte, D; Lambrecht, B; Lee, C-K; Marcelis, J; Marschner, S; McDonald, C; McGuane, S; McKee, M; Müller, T H; Muthivhi, T; Pettersson, A; Radziwon, P; Ramirez-Arcos, S; Reesink, H W; Rojo, J; Rood, I; Schmidt, M; Schneider, C K; Seifried, E; Sicker, U; Wendel, S; Wood, E M; Yomtovian, R A; Montag, T

2012-01-01

Bacterial contamination of platelet concentrates (PCs) still remains a significant problem in transfusion with potential important clinical consequences, including death. The International Society of Blood Transfusion Working Party on Transfusion-Transmitted Infectious Diseases, Subgroup on Bacteria, organised an international study on Transfusion-Relevant Bacteria References to be used as a tool for development, validation and comparison of both bacterial screening and pathogen reduction methods. Four Bacteria References (Staphylococcus epidermidis PEI-B-06, Streptococcus pyogenes PEI-B-20, Klebsiella pneumoniae PEI-B-08 and Escherichia coli PEI-B-19) were selected regarding their ability to proliferate to high counts in PCs and distributed anonymised to 14 laboratories in 10 countries for identification, enumeration and bacterial proliferation in PCs after low spiking (0·3 and 0·03 CFU/ml), to simulate contamination occurring during blood donation. Bacteria References were correctly identified in 98% of all 52 identifications. S. pyogenes and E. coli grew in PCs in 11 out of 12 laboratories, and K. pneumoniae and S. epidermidis replicated in all participating laboratories. The results of bacterial counts were very consistent between laboratories: the 95% confidence intervals were for S. epidermidis: 1·19-1·32 × 10(7) CFU/ml, S. pyogenes: 0·58-0·69 × 10(7) CFU/ml, K. pneumoniae: 18·71-20·26 × 10(7) CFU/ml and E. coli: 1·78-2·10 × 10(7) CFU/ml. The study was undertaken as a proof of principle with the aim to demonstrate (i) the quality, stability and suitability of the bacterial strains for low-titre spiking of blood components, (ii) the property of donor-independent proliferation in PCs, and (iii) their suitability for worldwide shipping of deep frozen, blinded pathogenic bacteria. These aims were successfully fulfilled. The WHO Expert Committee Biological Standardisation has approved the adoption of these four bacteria strains as the first Repository for Transfusion-Relevant Bacteria Reference Strains and, additionally, endorsed as a project the addition of six further bacteria strain preparations suitable for control of platelet contamination as the next step of enlargement of the repository. © 2011 The Author(s). Vox Sanguinis © 2011 International Society of Blood Transfusion.

Top–down effects of a grazing, omnivorous minnow ( Campostoma anomalum) on stream microbial communities

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

Veach, Allison M.; Troia, Matthew; Jumpponen, Ari

We report top–down control exerted by macroconsumers can strongly affect lower trophic levels and ecosystem processes. Studies of effects on primary consumers in streams have been focused on algae, and effects on bacteria are largely unknown. We manipulated the density of an omnivorous, grazing minnow, the central stoneroller (Campostoma anomalum), in experimental stream mesocosms (treatments with 0, 1, 2, 3, 4, 5, 6, or 7 individuals) to understand consumer effects on algal and bacterial abundance (chlorophyll a [Chl a] extraction, bacterial cell counts, biomass measurements) and bacterial diversity and community composition (via Illumina MiSeq sequencing of the V4 region ofmore » the 16S ribosomal RNA gene). Increasing C. anomalum density reduced algal biomass until density reached ~2 fish (5 g fish biomass/m 2), and higher fish densities did not affect algal biomass. Fish biomass did not affect bacterial cell counts. Biofilm organic matter decreased with increasing C. anomalum biomass. Bacterial community composition was not affected by fish biomass, but variation in community composition was correlated with shifts in bacterial abundances. Evenness of bacterial operational taxonomic units (OTUs) decreased with increasing C. anomalum biomass, indicating that bacterial communities exhibited a greater degree of OTU dominance when fish biomass was higher. These findings suggest that this grazing fish species reduces algal abundance and organic matter in low-nutrient streams until a threshold of moderate fish abundance is reached and that it reduces evenness of benthic bacterial communities but not bacterial biomass. Finally, given the importance of biofilm bacteria for ecosystem processes and the ubiquity of grazing fishes in streams, future researchers should explore both top–down and bottom–up interactions in alternative environmental contexts and with other grazing fish species.« less

Top–down effects of a grazing, omnivorous minnow ( Campostoma anomalum) on stream microbial communities

DOE PAGES

Veach, Allison M.; Troia, Matthew; Jumpponen, Ari; ...

2017-12-21

We report top–down control exerted by macroconsumers can strongly affect lower trophic levels and ecosystem processes. Studies of effects on primary consumers in streams have been focused on algae, and effects on bacteria are largely unknown. We manipulated the density of an omnivorous, grazing minnow, the central stoneroller (Campostoma anomalum), in experimental stream mesocosms (treatments with 0, 1, 2, 3, 4, 5, 6, or 7 individuals) to understand consumer effects on algal and bacterial abundance (chlorophyll a [Chl a] extraction, bacterial cell counts, biomass measurements) and bacterial diversity and community composition (via Illumina MiSeq sequencing of the V4 region ofmore » the 16S ribosomal RNA gene). Increasing C. anomalum density reduced algal biomass until density reached ~2 fish (5 g fish biomass/m 2), and higher fish densities did not affect algal biomass. Fish biomass did not affect bacterial cell counts. Biofilm organic matter decreased with increasing C. anomalum biomass. Bacterial community composition was not affected by fish biomass, but variation in community composition was correlated with shifts in bacterial abundances. Evenness of bacterial operational taxonomic units (OTUs) decreased with increasing C. anomalum biomass, indicating that bacterial communities exhibited a greater degree of OTU dominance when fish biomass was higher. These findings suggest that this grazing fish species reduces algal abundance and organic matter in low-nutrient streams until a threshold of moderate fish abundance is reached and that it reduces evenness of benthic bacterial communities but not bacterial biomass. Finally, given the importance of biofilm bacteria for ecosystem processes and the ubiquity of grazing fishes in streams, future researchers should explore both top–down and bottom–up interactions in alternative environmental contexts and with other grazing fish species.« less

Identity and Behavior of Xylem-Residing Bacteria in Rough Lemon Roots of Florida Citrus Trees †

PubMed Central

Gardner, John M.; Feldman, Albert W.; Zablotowicz, Robert M.

1982-01-01

An aseptic vacuum extraction technique was used to obtain xylem fluid from the roots of rough lemon (Citrus jambhiri Lush.) rootstock of Florida citrus trees. Bacteria were consistently isolated from vascular fluid of both healthy and young tree decline-affected trees. Thirteen genera of bacteria were found, the most frequently occurring genera being Pseudomonas (40%), Enterobacter (18%), Bacillus, Corynebacterium, and other gram-positive bacteria (16%), and Serratia (6%). Xylem bacterial counts fluctuated seasonally. Bacterial populations ranged from 0.1 to 22 per mm3 of root tissue (about 102 to 2 × 104 bacteria per g of xylem) when bacterial counts were made on vascular fluid, but these numbers were 10- to 1,000-fold greater when aseptically homogenized xylem tissue was examined similarly. Some of the resident bacteria (4%) are potentially phytopathogenic. It is proposed that xylem bacteria have an important role in the physiology of citrus. PMID:16346030

Food-safe modification of stainless steel food processing surfaces to reduce bacterial biofilms.

PubMed

Awad, Tarek Samir; Asker, Dalal; Hatton, Benjamin D

2018-06-11

Biofilm formation on stainless steel (SS) surfaces of food processing plants, leading to foodborne illness outbreaks, is enabled by the attachment and confinement within microscale cavities of surface roughness (grooves, scratches). We report Foodsafe Oil-based Slippery Coatings (FOSCs) for food processing surfaces that suppress bacterial adherence and biofilm formation by trapping residual oil lubricant within these surface cavities to block microbial growth. SS surfaces were chemically functionalized with alkylphosphonic acid to preferentially wet a layer of food grade oil. FOSCs reduced the effective surface roughness, the adhesion of organic food residue, and bacteria. FOSCs significantly reduced Pseudomonas aeruginosa biofilm formation on standard roughness SS-316 by 5 log CFU cm-2, and by 3 log CFU cm-2 for mirror-finished SS. FOSCs also enhanced surface cleanability, which we measured by bacterial counts after conventional detergent cleaning. Importantly, both SS grades maintained their anti-biofilm activity after erosion of the oil layer by surface wear with glass beads, which suggests there is a residual volume of oil that remains to block surface cavity defects. These results indicate the potential of such low-cost, scalable approaches to enhance the cleanability of SS food processing surfaces and improve food safety by reducing biofilm growth.

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.

Comparative evaluation of antibacterial property and substantivity of chlorhexidine containing dentifrices with sodium lauryl sulfate and Tween as surfactants: an in vivo study.

PubMed

Venu, V; Prabhakar, A R; Basappa, N

2013-01-01

The aim of the study was to determine the antibacterial property and substantivity of chlorhexidine containing dentifrices with sodium lauryl sulfate (SLS) and Tween as surfactants. It is a double-blind cross over the study, a total of 20 children within their mixed dentition period (7-13 year) having Streptococci mutans count more than 10(6) were selected for the main study. Three types of chlorhexidine containing dentifrices were used with a washout period of 1 week. Out of the three toothpastes, one was without surfactant and other two toothpastes contained SLS and Tween as surfactants respectively. 20 volunteers brushed for 1 min during the study day with their assigned toothpaste. Saliva samples were collected before brushing, immediately after brushing and 1, 3, 5, and 7 hand sent for microbial analysis. The culture carried out by inoculating saliva sample onto Mitis salivarius agar for selective isolation of S. mutans followed by counting of colony forming unit. Group I and III (Chlorhexidine and CHX + Tween) had shown statistically significant reduction in bacterial count until 7 h when compared to their baseline values ( P < 0.001). Group II toothpaste (CHX + SLS) had shown significant reduction in bacterial count until 3 h only. On inter group comparison, Group III had shown good amount of percentage reduction in bacterial count when compared to other groups. CHX + Tween toothpaste had shown statistically significant reduction in antibacterial activity and substantivity than other groups. These findings show chlorhexidine containing toothpaste with non-ionic surfactant will be able to maintain the antibacterial property and substantivity of chlorhexidine.

Pharmacodynamic Modeling of Bacillary Elimination Rates and Detection of Bacterial Lipid Bodies in Sputum to Predict and Understand Outcomes in Treatment of Pulmonary Tuberculosis

PubMed Central

Sloan, Derek J.; Mwandumba, Henry C.; Garton, Natalie J.; Khoo, Saye H.; Butterworth, Anthony E.; Allain, Theresa J.; Heyderman, Robert S.; Corbett, Elizabeth L.; Barer, Mike R.; Davies, Geraint R.

2015-01-01

Background. Antibiotic-tolerant bacterial persistence prevents treatment shortening in drug-susceptible tuberculosis, and accumulation of intracellular lipid bodies has been proposed to identify a persister phenotype of Mycobacterium tuberculosis cells. In Malawi, we modeled bacillary elimination rates (BERs) from sputum cultures and calculated the percentage of lipid body–positive acid-fast bacilli (%LB + AFB) on sputum smears. We assessed whether these putative measurements of persistence predict unfavorable outcomes (treatment failure/relapse). Methods. Adults with pulmonary tuberculosis received standard 6-month therapy. Sputum samples were collected during the first 8 weeks for serial sputum colony counting (SSCC) on agar and time-to positivity (TTP) measurement in mycobacterial growth indicator tubes. BERs were extracted from nonlinear and linear mixed-effects models, respectively, fitted to these datasets. The %LB + AFB counts were assessed by fluorescence microscopy. Patients were followed until 1 year posttreatment. Individual BERs and %LB + AFB counts were related to final outcomes. Results. One hundred and thirty-three patients (56% HIV coinfected) participated, and 15 unfavorable outcomes were reported. These were inversely associated with faster sterilization phase bacillary elimination from the SSCC model (odds ratio [OR], 0.39; 95% confidence interval [CI], .22–.70) and a faster BER from the TTP model (OR, 0.71; 95% CI, .55–.94). Higher %LB + AFB counts on day 21–28 were recorded in patients who suffered unfavorable final outcomes compared with those who achieved stable cure (P = .008). Conclusions. Modeling BERs predicts final outcome, and high %LB + AFB counts 3–4 weeks into therapy may identify a persister bacterial phenotype. These methods deserve further evaluation as surrogate endpoints for clinical trials. PMID:25778753

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.

[Assessment of the air quality improment of cleaning and disinfection on central air-conditioning ventilation system].

PubMed

Liu, Hongliang; Zhang, Lei; Feng, Lihong; Wang, Fei; Xue, Zhiming

2009-09-01

To assess the effect of air quality of cleaning and disinfection on central air-conditioning ventilation systems. 102 air-conditioning ventilation systems in 46 public facilities were sampled and investigated based on Hygienic assessment criterion of cleaning and disinfection of public central air-conditioning systems. Median dust volume decreased from 41.8 g/m2 to 0.4 g/m2, and the percentage of pipes meeting the national standard for dust decreased from 17.3% (13/60) to 100% (62/62). In the dust, median aerobic bacterial count decreased from 14 cfu/cm2 to 1 cfu/cm2. Median aerobic fungus count decreased from 10 cfu/cm2 to 0 cfu/cm2. The percentage of pipes with bacterial and fungus counts meeting the national standard increased from 92.4% (171/185) and 82.2% (152/185) to 99.4% (165/166) and 100% (166/166), respectively. In the ventilation air, median aerobic bacterial count decreased from 756 cfu/m3 to 229 cfu/m3. Median aerobic fungus count decreased from 382 cfu/m3 to 120 cfu/m3. The percentage of pipes meeting the national standard for ventilation air increased from 33.3% (81/243) and 62.1% (151/243) to 79.8% (292/366) and 87.7% (242/276), respectively. But PM10 rose from 0.060 mg/m3 to 0.068 mg/m3, and the percentage of pipes meeting the national standard for PM10 increased from 74.2% (13/60) to 90.2% (46/51). The cleaning and disinfection of central air-conditioning ventilation systems could have a beneficial effect of air quality.

Microbial contamination of cosmetics and personal care items in Egypt--body lotions and talcum powders.

PubMed

Ashour, M S; Abdelaziz, A A; Hefni, H; el-Tayeb, O M

1989-06-01

We examined a total of 54 samples, including 18 body lotions and 36 talcum powders, for their total aerobic bacterial, coliform and fungal counts. We also carried out anaerobic bacterial counts for talcum powder as well as tests to detect some potentially hazardous bacteria in all tested samples. Talcum powders were more heavily contaminated with bacteria and fungi than body lotions. More than 40% of the tested body lotions contained no viable bacteria or less than 100 c.f.u./g. while all the talcum powders tested contained more than 100 c.f.u./g. Thirty per cent of the talcum powders were contaminated with 10(4) c.f.u./g. and none of the body lotions were contaminated to that extent. No coliforms were recovered from any of the body lotions, while 17% of the talcum powder examined contained coliforms in the range of 230-500 c.f.u./g. Staphylococcus spp. were detected in 18 samples of both talcum powders and body lotions, three of these Staphylococci were of the aureus type. Three samples of talcum powder contained E. coli, two samples contained Enterobacter agglomerans and one sample contained Citrobacter freundii. Seventy per cent of the body lotions showed no fungal counts, while 83% of the talcum powders examined were contaminated with fungi and most of the contaminated talcum powders contained more than 100 fungal cells/g. With regard to the anaerobic bacterial counts for talcum powders, 50% of the samples showed no counts while the other 50% contained less than 100 c.f.u./g. Four samples were contaminated with Clostridium perfringens, although C. tetani was not recovered from any of the samples.

Influence of protein deposition on bacterial adhesion to contact lenses.

PubMed

Subbaraman, Lakshman N; Borazjani, Roya; Zhu, Hua; Zhao, Zhenjun; Jones, Lyndon; Willcox, Mark D P

2011-08-01

The aim of the study is to determine the adhesion of Gram positive and Gram negative bacteria onto conventional hydrogel (CH) and silicone hydrogel (SH) contact lens materials with and without lysozyme, lactoferrin, and albumin coating. Four lens types (three SH-balafilcon A, lotrafilcon B, and senofilcon A; one CH-etafilcon A) were coated with lysozyme, lactoferrin, or albumin (uncoated lenses acted as controls) and then incubated in Staphylococcus aureus (Saur 31) or either of two strains of Pseudomonas aeruginosa (Paer 6294 and 6206) for 24 h at 37 °C. The total counts of the adhered bacteria were determined using the H-thymidine method and viable counts by counting the number of colony-forming units on agar media. All three strains adhered significantly lower to uncoated etafilcon A lenses compared with uncoated SH lenses (p < 0.05). Lysozyme coating on all four lens types increased binding (total and viable counts) of Saur 31 (p < 0.05). However, lysozyme coating did not influence P. aeruginosa adhesion (p > 0.05). Lactoferrin coating on lenses increased binding (total and viable counts) of Saur 31 (p < 0.05). Lactoferrin-coated lenses showed significantly higher total counts (p < 0.05) but significantly lower viable counts (p < 0.05) of adhered P. aeruginosa strains. There was a significant difference between the total and viable counts (p < 0.05) that were bound to lactoferrin-coated lenses. Albumin coating of lenses increased binding (total and viable counts) of all three strains (p < 0.05). Lysozyme deposited on contact lenses does not possess antibacterial activity against certain bacterial strains, whereas lactoferrin possess an antibacterial effect against strains of P. aeruginosa.

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.

Microbial air quality in mass transport buses and work-related illness among bus drivers of Bangkok Mass Transit Authority.

PubMed

Luksamijarulkul, Pipat; Sundhiyodhin, Viboonsri; Luksamijarulkul, Soavalug; Kaewboonchoo, Orawan

2004-06-01

The air quality in mass transport buses, especially air-conditioned buses may affect bus drivers who work full time. Bus numbers 16, 63, 67 and 166 of the Seventh Bus Zone of Bangkok Mass Transit Authority were randomly selected to investigate for microbial air quality. Nine air-conditioned buses and 2-4 open-air buses for each number of the bus (36 air-conditioned buses and 12 open-air buses) were included. Five points of in-bus air samples in each studied bus were collected by using the Millipore A ir Tester Totally, 180 and 60 air samples collected from air-conditioned buses and open-air buses were cultured for bacterial and fungal counts. The bus drivers who drove the studied buses were interviewed towards histories of work-related illness while working. The results revealed that the mean +/- SD of bacterial counts in the studied open-air buses ranged from 358.50 +/- 146.66 CFU/m3 to 506 +/- 137.62 CFU/m3; bus number 16 had the highest level. As well as the mean +/- SD of fungal counts which ranged from 93.33 +/- 44.83 CFU/m3 to 302 +/- 294.65 CFU/m3; bus number 166 had the highest level. Whereas, the mean +/- SD of bacterial counts in the studied air-conditioned buses ranged from 115.24 +/- 136.01 CFU/m3 to 244.69 +/- 234.85 CFU/m3; bus numbers 16 and 67 had the highest level. As well as the mean +/- SD of fungal counts which rangedfrom 18.84 +/- 39.42 CFU/m3 to 96.13 +/- 234.76 CFU/m3; bus number 166 had the highest level. When 180 and 60 studied air samples were analyzed in detail, it was found that 33.33% of the air samples from open-air buses and 6.11% of air samples from air-conditioned buses had a high level of bacterial counts (> 500 CFU/m3) while 6.67% of air samples from open-air buses and 2.78% of air samples from air-conditioned buses had a high level of fungal counts (> 500 CFU/m3). Data from the history of work-related illnesses among the studied bus drivers showed that 91.67% of open-air bus drivers and 57.28% of air-conditioned bus drivers had symptoms of work-related illnesses, p = 0.0185.

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.

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.

Neutrophil Extracellular Traps and Fibrin in Otitis Media: Analysis of Human and Chinchilla Temporal Bones.

PubMed

Schachern, Patricia A; Kwon, Geeyoun; Briles, David E; Ferrieri, Patricia; Juhn, Steven; Cureoglu, Sebahattin; Paparella, Michael M; Tsuprun, Vladimir

2017-10-01

Bacterial resistance in acute otitis can result in bacterial persistence and biofilm formation, triggering chronic and recurrent infections. To investigate the middle ear inflammatory response to bacterial infection in human and chinchilla temporal bones. Six chinchillas underwent intrabullar inoculations with 0.5 mL of 106 colony-forming units (CFUs) of Streptococcus pneumoniae, serotype 2. Two days later, we counted bacteria in middle ear effusions postmortem. One ear from each chinchilla was processed in paraffin and sectioned at 5 µm. The opposite ear was embedded in epoxy resin, sectioned at a thickness of 1 µm, and stained with toluidine blue. In addition, we examined human temporal bones from 2 deceased donors with clinical histories of otitis media (1 with acute onset otitis media, 1 with recurrent infection). Temporal bones had been previously removed at autopsy, processed, embedded in celloidin, and cut at a thickness of 20 µm. Sections of temporal bones from both chinchillas and humans were stained with hematoxylin-eosin and immunolabeled with antifibrin and antihistone H4 antibodies. Histopatological and imminohistochemical changes owing to otitis media. Bacterial counts in chinchilla middle ear effusions 2 days after inoculation were approximately 2 logs above initial inoculum counts. Both human and chinchilla middle ear effusions contained bacteria embedded in a fibrous matrix. Some fibers in the matrix showed positive staining with antifibrin antibody, others with antihistone H4 antibody. In acute and recurrent otitis media, fibrin and neutrophil extracellular traps (NETs) are part of the host inflammatory response to bacterial infection. In the early stages of otitis media the host defense system uses fibrin to entrap bacteria, and NETs function to eliminate bacteria. In chronic otitis media, fibrin and NETs appear to persist.

Specific MAIT cell behaviour among innate-like T lymphocytes in critically ill patients with severe infections.

PubMed

Grimaldi, David; Le Bourhis, Lionel; Sauneuf, Bertrand; Dechartres, Agnès; Rousseau, Christophe; Ouaaz, Fatah; Milder, Maud; Louis, Delphine; Chiche, Jean-Daniel; Mira, Jean-Paul; Lantz, Olivier; Pène, Frédéric

2014-02-01

In between innate and adaptive immunity, the recently identified innate-like mucosal-associated invariant T (MAIT) lymphocytes display specific reactivity to non-streptococcal bacteria. Whether they are involved in bacterial sepsis has not been investigated. We aimed to assess the number and the time course of circulating innate-like T lymphocytes (MAIT, NKT and γδ T cells) in critically ill septic and non-septic patients and to establish correlations with the further development of intensive care unit (ICU)-acquired infections. We prospectively enrolled consecutive patients with severe sepsis and septic shock. Controls were critically ill patients with non-septic shock and age-matched healthy subjects. Circulating innate-like lymphocytes were enumerated using a flow cytometry assay at day 1, 4 and 7. One hundred and fifty six patients (113 severe bacterial infections, 36 non-infected patients and 7 patients with severe viral infections) and 26 healthy subjects were enrolled into the study. Patients with severe bacterial infections displayed an early decrease in MAIT cell count [median 1.3/mm(3); interquartile range (0.4-3.2)] as compared to control healthy subjects [31.1/mm(3) (12.1-45.2)], but also to non-infected critically ill patients [4.3/mm(3) (1.4-13.2)] (P < 0.0001 for all comparisons). In contrast NKT and γδ T cell counts did not differ between patients groups. The multivariate analysis identified non-streptococcal bacterial infection as an independent determinant of decrease in MAIT cell count. Furthermore, the incidence of ICU-acquired infections was higher in patients with persistent MAIT cell depletion. This large human study provides valuable information about MAIT cells in severe bacterial infections. The persistent depletion of MAIT cells is associated with the further development of ICU-acquired infections.

Rationale behind high-dose amoxicillin therapy for acute otitis media due to penicillin-nonsusceptible pneumococci: support from in vitro pharmacodynamic studies.

PubMed Central

Lister, P D; Pong, A; Chartrand, S A; Sanders, C C

1997-01-01

To evaluate whether increased doses of amoxicillin should be used to treat acute pneumococcal otitis media, an in vitro pharmacokinetic model was used to evaluate the killing of pneumococci by amoxicillin when middle ear pharmacokinetics were simulated. Logarithmic-phase cultures were exposed to peak concentrations of 3, 6, and 9 microg of amoxicillin per ml every 12 h, and an elimination half-life of 1.6 h was simulated. Changes in viable bacterial counts were measured over 36 h. All three doses rapidly decreased the viable bacterial counts of penicillin-susceptible strains below the 10-CFU/ml limit of detection by 6 to 10 h and maintained counts below this limit through 36 h. The 3-microg/ml peak dose was much less effective against two of three strains with intermediate penicillin resistance and all three penicillin-resistant strains, with bacterial counts approaching those in drug-free control cultures by 12 h. The 6-microg/ml peak dose completely eliminated two of three strains with intermediate penicillin resistance and maintained viable counts of the other nonsusceptible strains at 1.5 to 2 logs below the initial inoculum through 36 h. The 9-microg/ml peak dose was most effective, completely eliminating all three strains with intermediate penicillin resistance and maintaining the viable counts of the resistant strains at 3 to 4 logs below the original inoculum. The pharmacodynamics observed in this study suggest that peak concentrations of amoxicillin of 6 to 9 microg/ml may be sufficient for the elimination of penicillin-nonsusceptible pneumococcal strains causing otitis media, especially those with intermediate resistance to amoxicillin. In vivo pharmacokinetic studies are needed to determine if these levels can be achieved in middle ear fluid with amoxicillin at 70 to 90 mg/kg/day divided into two daily doses. If these levels are reliably achieved, then clinical studies are warranted. PMID:9303386

Methods for quantitative and qualitative evaluation of vaginal microflora during menstruation.

PubMed Central

Onderdonk, A B; Zamarchi, G R; Walsh, J A; Mellor, R D; Muñoz, A; Kass, E H

1986-01-01

The quantitative and qualitative changes in the bacterial flora of the vagina during menstruation have received inadequate study. Similarly, the effect of vaginal tampons on the microbial flora as well as the relationship between the microbial flora of the vagina and that of the tampon has not been adequately evaluated. The purposes of the present study were (i) to develop quantitative methods for studying the vaginal flora and the flora of tampons obtained during menstruation and (ii) to determine whether there were differences between the microflora of the tampon and that of the vaginal vault. Tampon and swab samples were obtained at various times from eight young healthy volunteers for 8 to 10 menstrual cycles. Samples consisted of swabs from women wearing menstrual pads compared with swab and tampon samples taken at various times during the menstrual cycle. Samples were analyzed for total facultative and anaerobic bacterial counts, and the six dominant bacterial species in each culture were identified. Statistical evaluation of the results indicates that total bacterial counts decreased during menstruation and that swab and tampon samples yielded similar total counts per unit weight of sample. The numbers of bacteria in tampons tended to be lower than in swabs taken at the same time. Overall, during menstruation, the concentrations of lactobacilli declined, but otherwise there was little difference among the species found during menstruation compared with those found in intermenstrual samples. Cotton tampons had little discernible effect on the microbial flora. PMID:3954346

Evaluation in a Dog Model of Three Antimicrobial Glassy Coatings: Prevention of Bone Loss around Implants and Microbial Assessments

PubMed Central

López-Píriz, Roberto; Solá-Linares, Eva; Rodriguez-Portugal, Mercedes; Malpica, Beatriz; Díaz-Güemes, Idoia; Enciso, Silvia; Esteban-Tejeda, Leticia; Cabal, Belén; Granizo, Juan José; Moya, José Serafín; Torrecillas, Ramón

2015-01-01

Objectives The aim of the present study is to evaluate, in a ligature-induced peri-implantitis model, the efficacy of three antimicrobial glassy coatings in the prevention of biofilm formation, intrasulcular bacterial growth and the resulting peri-implant bone loss. Methods Mandibular premolars were bilaterally extracted from five beagle dogs. Four dental implants were inserted on each hemiarch. Eight weeks after, one control zirconia abutment and three with different bactericidal coatings (G1n-Ag, ZnO35, G3) were connected. After a plaque control period, bacterial accumulation was allowed and biofilm formation on abutments was observed by Scanning Electron Microscopy (SEM). Peri-implantitis was induced by cotton ligatures. Microbial samples and peri-implant crestal bone levels of all implant sites were obtained before, during and after the breakdown period. Results During experimental induce peri-implantitis: colony forming units counts from intrasulcular microbial samples at implants with G1n-Ag coated abutment remained close to the basal inoculum; G3 and ZnO35 coatings showed similar low counts; and anaerobic bacterias counts at control abutments exhibited a logarithmic increase by more than 2. Bone loss during passive breakdown period was no statistically significant. Additional bone loss occurred during ligature-induce breakdown: 0.71 (SD 0.48) at G3 coating, 0.57 (SD 0.36) at ZnO35 coating, 0.74 (SD 0.47) at G1n-Ag coating, and 1.29 (SD 0.45) at control abutments; and statistically significant differences (p<0.001) were found. The lowest bone loss at the end of the experiment was exhibited by implants dressing G3 coated abutments (mean 2.1; SD 0.42). Significance Antimicrobial glassy coatings could be a useful tool to ward off, diminish or delay peri-implantitis progression. PMID:26489088

Survival of foodborne pathogenic bacteria (Bacillus cereus, Escherichia coli O157:H7, Salmonella enterica serovar Typhimurium, Staphylococcus aureus, and Listeria monocytogenes) and Bacillus cereus spores in fermented alcoholic beverages (beer and refined rice wine).

PubMed

Kim, S A; Kim, N H; Lee, S H; Hwang, I G; Rhee, M S

2014-03-01

Only limited information is available on the microbiological safety of fermented alcoholic beverages because it is still a common belief that such beverages do not provide a favorable environment for bacterial growth and survival. Thus, in this study, we examined the survival of major foodborne pathogens and spores in fermented alcoholic beverages. Foodborne pathogens (Bacillus cereus, Escherichia coli O157:H7, Listeria monocytogenes, Salmonella enterica serovar Typhimurium, and Staphylococcus aureus) and B. cereus spores (initial population, 3 to 4 log CFU/ml) were inoculated separately into three types of beer and refined rice wine, which were then stored at 5 and 22°C. Bacterial counts were assayed periodically for up to 28 days. Vegetative B. cereus counts decreased rapidly, whereas B. cereus spore counts remained constant (P > 0.05) for a long period of time in all beverages. Vegetative B. cereus cells formed spores in beer at 5 and 22°C, and the spores survived for long periods. Among vegetative cells, E. coli O157:H7 had the highest survival (only 1.49 to 1.56 log reduction during 28 days in beer at 5°C). Beer and refined rice wine supported microbial survival from several days to several weeks. Our results appear to contradict the common belief that pathogens cannot survive in alcoholic beverages. Long-term survival of pathogens (especially B. cereus and E. coli O157:H7) in beer and refined rice wine should be taken into consideration by the manufacturers of these beverages. This study provides basic information that should help further research into microbial survival in alcoholic beverages and increase the microbiological safety regulation of fermented alcoholic beverages.

Effect of human milk fortifiers on bacterial growth in human milk.

PubMed

Santiago, Myla S; Codipilly, Champa N; Potak, Debra C; Schanler, Richard J

2005-10-01

As a component in human milk fortifiers (HMF), iron may equilibrate with human milk for as long as 24 hours, bind important bacteriostatic proteins, and potentially affect the host defense properties of human milk. We compared bacterial growth in human milk prepared with each of two HMF differing in their content of iron. Samples of human milk obtained from mothers of premature infants were divided and mixed with one of two HMF and maintained at refrigerator temperature. Refrigerated milk samples were removed at 0, 24, and 72 hours for determination of total bacterial colony counts (TBCC). TBCC did not differ between groups but declined from 0 to 72 hours, p<0.001. These data suggest that differences in iron content, or other nutrients in HMF, do not affect bacterial growth in human milk. Storage of fortified human milk at refrigerator temperature for 72 hours results in decreased bacterial growth. As a component in human milk fortifiers (HMF), iron may equilibrate with human milk for as long as 24 hours, bind important bacteriostatic proteins, and potentially affect the host defense properties of human milk. We compared bacterial growth in human milk prepared with each of two HMF differing in their content of iron. Samples of human milk obtained from mothers of premature infants were divided and mixed with one of two HMF and maintained at refrigerator temperature. Refrigerated milk samples were removed at 0, 24, and 72 hours for determination of total bacterial colony counts (TBCC).

Influence of Thawing Methods and Storage Temperatures on Bacterial Diversity, Growth Kinetics, and Biogenic Amine Development in Atlantic Mackerel.

PubMed

Onyango, S; Palmadottir, H; Tómason, T; Marteinsson, V T; Njage, P M K; Reynisson, E

2016-11-01

Limited knowledge is currently available on the influence of fish thawing and subsequent storage conditions on bacterial growth kinetics, succession, and diversity alongside the production of biogenic amines. This study aimed to address these factors during the thawing and subsequent storage of mackerel. Thawing was either done fast in 18°C water for 2 h or slowly at 30°C overnight. Subsequent storage was at 30°C (ambient) for 36 h and 2 to 5°C (refrigerated) for 12 days. The cultivation methods used were total viable counts, hydrogen sulfide-producing bacteria, and Pseudomonas . Maximum growth rate, population density, and lag time were fitted on the counts using the Baranyi model. The bacterial diversity and succession were based on sequencing of 16S rRNA amplicons, and biogenic amines were quantified on high-pressure liquid chromatography-UV. The results show that lag time of hydrogen sulfide-producing bacteria was significantly affected by both thawing methods, and further, the interaction between thawing and storage significantly affected the maximum growth rate of these bacteria. However, the maximum growth rate of Pseudomonas was higher during refrigerated storage compared with storage at ambient temperature. Total viable counts showed longer lag time and reduced growth rate under refrigerated storage. Higher bacterial diversity was correlated to slow thawing and storage at ambient temperature compared with slow thawing and refrigerated storage. Overall, Acinetobacter and Psychrobacter genera were the dominant bacterial populations. The amine levels were low and could not be differentiated along the thawing and storage approaches, despite a clear increase in bacterial load, succession, and diversity. This corresponded well with the low abundance of biogenic amine-producing bacteria, with the exception of the genus Proteus , which was 8.6% in fast-thawed mackerel during storage at ambient temperature. This suggests that the decarboxylation potential is dependent on both microbial load and microbial community structure.

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

PubMed

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

2015-04-01

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

Clinical antibacterial effectiveness of the self-adjusting file system.

PubMed

Neves, M A S; Rôças, I N; Siqueira, J F

2014-04-01

To evaluate in vivo the antibacterial effectiveness of the self-adjusting file (SAF) using molecular methods. Root canals from single-rooted teeth with apical periodontitis were instrumented using the SAF system under continuous irrigation with 2.5% NaOCl. DNA extracts from samples taken before and after instrumentation were subjected to quantitative analysis of total bacteria counts and levels of streptococci by quantitative real-time polymerase chain reaction (qPCR). The reverse-capture checkerboard assay was also used to identify 28 bacterial taxa before (S1) and after (S2) SAF instrumentation. SAF was also compared with a conventional hand nickel-titanium instrumentation technique for total bacterial reduction. Data from qPCR were analysed statistically within groups using the Wilcoxon matched pairs test and between groups using the Mann-Whitney U-test and the Fisher's exact test, with significance level set at P < 0.05. Self-adjusting file significantly reduced the total bacterial counts from a mean number of 1.96 × 10(7) cells to 1.34 × 10(4) cells (P < 0.001). Quantitatively, the 99.9% reduction in total bacterial counts associated with the SAF system was significantly superior to the 95.1% reduction obtained by hand instrumentation (P < 0.001). Qualitatively, SAF resulted in significantly more cases with negative PCR results for bacteria (54.5%) than hand instrumentation (4.5%) (P < 0.001). The SAF system succeeded in significantly reducing the streptococcal levels, but four cases still harboured these bacteria in S2. Checkerboard analysis revealed that not only streptococci but also some anaerobic and even as-yet-uncultivated bacteria may resist the effects of chemomechanical procedures. The SAF instrumentation system was highly effective in reducing bacterial populations from infected root canals and performed significantly better than hand instrumentation. However, because half of the samples still had detectable bacteria after preparation with SAF, supplementary disinfection is still required to maximize bacterial elimination. © 2013 International Endodontic Journal. Published by John Wiley & Sons Ltd.

Diagnostic value of lactate, procalcitonin, ferritin, serum-C-reactive protein, and other biomarkers in bacterial and viral meningitis: A cross-sectional study.

PubMed

Sanaei Dashti, Anahita; Alizadeh, Shekoofan; Karimi, Abdullah; Khalifeh, Masoomeh; Shoja, Seyed Abdolmajid

2017-09-01

There are many difficulties distinguishing bacterial from viral meningitis that could be reasonably solved using biomarkers. The aim of this study was to evaluate lactate, procalcitonin (PCT), ferritin, serum-CRP (C-reactive protein), and other known biomarkers in differentiating bacterial meningitis from viral meningitis in children.All children aged 28 days to 14 years with suspected meningitis who were admitted to Mofid Children's Hospital, Tehran, between October 2012 and November 2013, were enrolled in this prospective cross-sectional study. Children were divided into 2 groups of bacterial and viral meningitis, based on the results of cerebrospinal fluid (CSF) culture, polymerase chain reaction, and cytochemical profile. Diagnostic values of CSF parameters (ferritin, PCT, absolute neutrophil count [ANC], white blood cell count, and lactate) and serum parameters (PCT, ferritin, CRP, and erythrocyte sedimentation rate [ESR]) were evaluated.Among 50 patients with meningitis, 12 were diagnosed with bacterial meningitis. Concentrations of all markers were significantly different between bacterial and viral meningitis, except for serum (P = .389) and CSF (P = .136) PCT. The best rates of area under the receiver operating characteristic (ROC) curve (AUC) were achieved by lactate (AUC = 0.923) and serum-CRP (AUC = 0.889). The best negative predictive values (NPV) for bacterial meningitis were attained by ANC (100%) and lactate (97.1%).The results of our study suggest that ferritin and PCT are not strong predictive biomarkers. A combination of low CSF lactate, ANC, ESR, and serum-CRP could reasonably rule out the bacterial meningitis.

Diagnostic value of lactate, procalcitonin, ferritin, serum-C-reactive protein, and other biomarkers in bacterial and viral meningitis

PubMed Central

Sanaei Dashti, Anahita; Alizadeh, Shekoofan; Karimi, Abdullah; Khalifeh, Masoomeh; Shoja, Seyed Abdolmajid

2017-01-01

Abstract There are many difficulties distinguishing bacterial from viral meningitis that could be reasonably solved using biomarkers. The aim of this study was to evaluate lactate, procalcitonin (PCT), ferritin, serum-CRP (C-reactive protein), and other known biomarkers in differentiating bacterial meningitis from viral meningitis in children. All children aged 28 days to 14 years with suspected meningitis who were admitted to Mofid Children's Hospital, Tehran, between October 2012 and November 2013, were enrolled in this prospective cross-sectional study. Children were divided into 2 groups of bacterial and viral meningitis, based on the results of cerebrospinal fluid (CSF) culture, polymerase chain reaction, and cytochemical profile. Diagnostic values of CSF parameters (ferritin, PCT, absolute neutrophil count [ANC], white blood cell count, and lactate) and serum parameters (PCT, ferritin, CRP, and erythrocyte sedimentation rate [ESR]) were evaluated. Among 50 patients with meningitis, 12 were diagnosed with bacterial meningitis. Concentrations of all markers were significantly different between bacterial and viral meningitis, except for serum (P = .389) and CSF (P = .136) PCT. The best rates of area under the receiver operating characteristic (ROC) curve (AUC) were achieved by lactate (AUC = 0.923) and serum-CRP (AUC = 0.889). The best negative predictive values (NPV) for bacterial meningitis were attained by ANC (100%) and lactate (97.1%). The results of our study suggest that ferritin and PCT are not strong predictive biomarkers. A combination of low CSF lactate, ANC, ESR, and serum-CRP could reasonably rule out the bacterial meningitis. PMID:28858084

Histological observations in the Hawaiian reef coral, Porites compressa, affected by Porites bleaching with tissue loss.

PubMed

Sudek, M; Work, T M; Aeby, G S; Davy, S K

2012-10-01

The scleractinian finger coral Porites compressa is affected by the coral disease Porites bleaching with tissue loss (PBTL). This disease initially manifests as bleaching of the coenenchyme (tissue between polyps) while the polyps remain brown with eventual tissue loss and subsequent algal overgrowth of the bare skeleton. Histopathological investigation showed a loss of symbiont and melanin-containing granular cells which was more pronounced in the coenenchyme than the polyps. Cell counts confirmed a 65% reduction in symbiont density. Tissue loss was due to tissue fragmentation and necrosis in affected areas. In addition, a reduction in putative bacterial aggregate densities was found in diseased samples but no potential pathogens were observed. Copyright © 2012 Elsevier Inc. All rights reserved.

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

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.

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.

Microbial aerosol liberation from soiled textiles isolated during routine residuals handling in a modern health care setting.

PubMed

Handorean, Alina; Robertson, Charles E; Harris, J Kirk; Frank, Daniel; Hull, Natalie; Kotter, Cassandra; Stevens, Mark J; Baumgardner, Darrel; Pace, Norman R; Hernandez, Mark

2015-12-09

A wide variety of specialty textiles are used in health care settings for bedding, clothing, and privacy. The ability of textiles to host or otherwise sequester microbes has been well documented; however, their reciprocal potential for liberating airborne bacteria remains poorly characterized. In response, a multi-season survey of bacterial bioaerosols was conducted in the origin and terminus of residual paths which are specifically designed to isolate soiled hospital textiles as they are moved to laundering. This survey used conventional optical particle counting which incorporated multi-channel fluorescence in conjunction with molecular phylogenetic analyses to characterize the bioaerosols liberated during soiled textile storage--immediately before and after the occupation of a modern hospital. Although outfitted with a HEPA filtration system, the number of airborne particles presenting fluorescing optical signatures consistent with airborne bacteria and fungi significantly increased in textile holding rooms soon after the hospital's commissioning, even though these isolated residual areas rarely host personnel. The bioaerosol liberated during textile storage was characterized using Illumina MiSeq sequencing of bacterial 16S ribosomal ribonucleic acid (rRNA) genes. Gene copies recovered by quantitative PCR from aerosol collected in co-located impingers were consistent with fluorescence gated optical particle counting. The relative abundance patterns of proximal bacterial bioaerosol were such that the air in the origin and terminus of textile storage rooms could not be differentiated once the hospital began processing soiled linens. Genes from microbes typically associating with human skin, feces, and hair--Staphylococcus, Propionibacteria, Corynebacteria, Lactobacillus, and Streptococcus spp.--dominated the aerosol abundance profiles in textile holding rooms, which were generally far less diverse than communities recovered from surfaces in patient rooms. These results suggest that aerosol partitioning from the routine handling of soiled textiles can contribute to airborne exposures in the health care environment.

Purifying, Separating, and Concentrating Cells From a Sample Low in Biomass

NASA Technical Reports Server (NTRS)

Benardini, James N.; LaDuc, Myron T.; Diamond, Rochelle

2012-01-01

Frequently there is an inability to process and analyze samples of low biomass due to limiting amounts of relevant biomaterial in the sample. Furthermore, molecular biological protocols geared towards increasing the density of recovered cells and biomolecules of interest, by their very nature, also concentrate unwanted inhibitory humic acids and other particulates that have an adversarial effect on downstream analysis. A novel and robust fluorescence-activated cell-sorting (FACS)-based technology has been developed for purifying (removing cells from sampling matrices), separating (based on size, density, morphology), and concentrating cells (spores, prokaryotic, eukaryotic) from a sample low in biomass. The technology capitalizes on fluorescent cell-sorting technologies to purify and concentrate bacterial cells from a low-biomass, high-volume sample. Over the past decade, cell-sorting detection systems have undergone enhancements and increased sensitivity, making bacterial cell sorting a feasible concept. Although there are many unknown limitations with regard to the applicability of this technology to environmental samples (smaller cells, few cells, mixed populations), dogmatic principles support the theoretical effectiveness of this technique upon thorough testing and proper optimization. Furthermore, the pilot study from which this report is based proved effective and demonstrated this technology capable of sorting and concentrating bacterial endospore and bacterial cells of varying size and morphology. Two commercial off-the-shelf bacterial counting kits were used to optimize a bacterial stain/dye FACS protocol. A LIVE/DEAD BacLight Viability and Counting Kit was used to distinguish between the live and dead cells. A Bacterial Counting Kit comprising SYTO BC (mixture of SYTO dyes) was employed as a broad-spectrum bacterial counting agent. Optimization using epifluorescence microscopy was performed with these two dye/stains. This refined protocol was further validated using varying ratios and mixtures of cells to ensure homogenous staining compared to that of individual cells, and were utilized for flow analyzer and FACS labeling. This technology focuses on the purification and concentration of cells from low-biomass spacecraft assembly facility samples. Currently, purification and concentration of low-biomass samples plague planetary protection downstream analyses. Having a capability to use flow cytometry to concentrate cells out of low-biomass, high-volume spacecraft/ facility sample extracts will be of extreme benefit to the fields of planetary protection and astrobiology. Successful research and development of this novel methodology will significantly increase the knowledge base for designing more effective cleaning protocols, and ultimately lead to a more empirical and true account of the microbial diversity present on spacecraft surfaces. Refined cleaning and an enhanced ability to resolve microbial diversity may decrease the overall cost of spacecraft assembly and/or provide a means to begin to assess challenging planetary protection missions.

Cellular and soluble components decrease the viable pathogen counts in milk from dairy cows with subclinical mastitis.

PubMed

Koshiishi, Tomoko; Watanabe, Masako; Miyake, Hajime; Hisaeda, Keiichi; Isobe, Naoki

2017-08-10

The present study was undertaken to clarify the factors that reduce the viable pathogen count in milk collected from the udders of subclinical mastitic cows during preservation. Milk was centrifuged to divide somatic cells (cellular components, precipitates) and antimicrobial peptides (soluble components, supernatants without fat layer); each fraction was cultured with bacteria, and the number of viable bacteria was assessed prior to and after culture. In 28.8% of milk samples, we noted no viable bacteria immediately after collection; this value increased significantly after a 5-hr incubation of milk with cellular components but not with soluble components (48.1 and 28.8%, respectively). After culture with cellular components, the numbers of bacteria (excluding Staphylococcus aureus and Streptococcus uberis) and yeast decreased dramatically, although the differences were not statistically significant. After cultivation with soluble components, only yeasts showed a tendency toward decreased mean viability, whereas the mean bacterial counts of S. uberis and T. pyogenes tended to increase after 5-hr preservation with soluble components. These results suggest that most pathogens in high somatic cell count (SCC) milk decreased during preservation at 15 to 25°C, due to both the cellular components and antimicrobial components in the milk. Particularly, the cellular components more potently reduced bacterial counts during preservation.

Refrigerator storage of expressed human milk in the neonatal intensive care unit.

PubMed

Slutzah, Meredith; Codipilly, Champa N; Potak, Debra; Clark, Richard M; Schanler, Richard J

2010-01-01

To provide recommendations for refrigerator storage of human milk, the overall integrity (bacterial growth, cell counts, and component concentrations) of milk was examined during 96 hours of storage at 4 degrees C. Fresh milk samples (n = 36) were divided and stored at 4 degrees C for 0, 24, 48, 72, and 96 hours. At each time, pH, white cell count, and osmolality were measured and additional samples were stored at -80 degrees C until analyzed for bacteria and concentrations of lactoferrin, secretory (s)IgA, fat, fatty acids, and protein. There were no significant changes for osmolality, total and Gram-negative bacterial colony counts or concentrations of sIgA, lactoferrin, and fat. Gram-positive colony counts (2.9 to 1.6 x 10(5) colony-forming units per mL), pH (7.21 to 6.68), white blood cell counts (2.31 to 1.85 x 10(6) cells per mL), and total protein (17.5 to 16.7 g/L) declined, and free fatty acid concentrations increased (0.35 to 1.28 g/L) as storage duration increased, P < .001. Changes were minimal and the overall integrity of milk during refrigerator storage was preserved. Fresh mother's milk may be stored at refrigerator temperature for as long as 96 hours.

Contribution of bacterial cells to lacustrine organic matter based on amino sugars and D-amino acids

NASA Astrophysics Data System (ADS)

Carstens, Dörte; Köllner, Krista E.; Bürgmann, Helmut; Wehrli, Bernhard; Schubert, Carsten J.

2012-07-01

Amino sugars (ASs), D-amino acids (D-AAs), and bacterial cell counts were measured in two Swiss lakes to study the contribution of bacterial cells to organic matter (OM) and the fate of ASs and bacterial amino biomarkers during OM degradation. Concentrations of individual ASs (glucosamine, galactosamine, muramic acid, and mannosamine) in the particulate and total OM pools were analyzed in water-column profiles of Lake Brienz (oligotrophic and oxic throughout the entire water column) and Lake Zug (eutrophic, stratified, and permanently anoxic below 170 m) in spring and in fall. Generally, carbon-normalized AS concentrations decreased with water depth, indicating the preferential decomposition of ASs. For Lake Brienz the relative loss of particulate ASs was higher than in Lake Zug, suggesting enhanced AS turnover in an oligotrophic environment. AS ratio changes in the water column revealed a replacement of plankton biomass with OM from heterotrophic microorganisms with increasing water depth. Similar to the ASs, highest carbon normalized D-AA concentrations were found in the upper water column with decreasing concentrations with depth and an increase close to the sediments. In Lake Zug, an increase in the percentage of D-AAs also showed the involvement of bacteria in OM degradation. Estimations of OM derived from bacterial cells using cell counts and the bacterial biomarkers muramic acid and D-AAs gave similar results. For Lake Brienz 0.2-14% of the organic carbon pool originated from bacterial cells, compared to only 0.1-5% in Lake Zug. Based on our estimates, muramic acid appeared primarily associated with bacterial biomass and not with refractory bacterial necromass. Our study underscores that bacteria are not only important drivers of OM degradation in lacustrine systems, they also represent a significant source of OM themselves, especially in oligotrophic lakes.

In vitro adherence of radioactively labeled Escherichia coli in normal and cystitis-prone females

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

Parsons, C.L.; Anwar, H.; Stauffer, C.

Numerous investigators report data obtained using an in vitro quantitative assay for measuring bacterial adherence to epithelial cells. In the modified assay described here, we eliminated the need for visual counting of bacteria by incorporating the use of radioactively labeled Escherichia coli. This allowed quantitation of bacterial adherence to as many as 50,000 vaginal cells, whereas the visual counting system limits the determination to perhaps 50 cells. Using the modified method, we found no statistically significant differences among values for adherence of E. coli type 04 to the vaginal cells of control and cystitis-prone women at either pH 6.4 ormore » 4.0.« less

Bacteriological status of beef carcasses at a commercial abattoir before and after slaughterline improvements.

PubMed Central

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

1987-01-01

The bacteriological status of beef carcasses was monitored at a commercial abattoir before and after two stages of modernization to the beef slaughterline which included changing from cradle dressing to dressing on an overhead rail, and the introduction of hot water spray cleaning of carcasses. Although small significant (P less than 0.05) differences in bacterial count occurred among carcass sites within modernization stages, significant visit within stage variation and stage X site interactions prevented any significant change in overall count being observed among stages and carcass sites. Principal components analysis revealed small changes in the distribution of bacterial numbers on the sites sampled. PMID:3556439

Pilot-scale biopesticide production by Bacillus thuringiensis subsp. kurstaki using starch industry wastewater as raw material.

PubMed

Ndao, Adama; Sellamuthu, Balasubramanian; Gnepe, Jean R; Tyagi, Rajeshwar D; Valero, Jose R

2017-09-02

Pilot-scale Bacillus thuringiensis based biopesticide production (2000 L bioreactor) was conducted using starch industry wastewater (SIW) as a raw material using optimized operational parameters obtained in 15 L and 150 L fermenters. In pilot scale fermentation process the oxygen transfer rate is a major limiting factor for high product yield. Thus, the volumetric mass transfer coefficient (K L a) remains a tool to determine the oxygen transfer capacity [oxygen utilization rate (OUR) and oxygen transfer rate (OTR)] to obtain better bacterial growth rate and entomotoxicity in new bioreactor process optimization and scale-up. This study results demonstrated that the oxygen transfer rate in 2000 L bioreactor was better than 15 L and 150 L fermenters. The better oxygen transfer in 2000 L bioreactor augmented the bacterial growth [total cell (TC) and viable spore count (SC)] and delta-endotoxin yield. Prepared a stable biopesticide formulation for field use and its entomotoxicity was also evaluated. This study result corroborates the feasibility of industrial scale operation of biopesticide production using starch industry wastewater as raw material.

Associations between management practices and colostrum quality on New Zealand dairy farms.

PubMed

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

2017-09-01

To describe colostrum quality in spring-calving dairy herds in New Zealand, in terms of Brix, pH and total and coliform bacterial counts and to investigate associations between farm management practices and these measures of colostrum quality. In June 2015, commercial dairy farms (n=105), located in North and South Islands of New Zealand, were visited shortly after the first cows had calved, and when approximately 50% and 80% of the herd had calved (early, middle and late visits). One litre of pooled colostrum that was being fed to newborn calves was collected at each visit and used to determine Brix, pH, total bacterial and coliform counts. A survey of calf management practices was conducted with the herd manager or calf rearer after the final visit. Of 298 pooled colostrum samples tested 29/298 (9.7%) had Brix >22%. Brix was higher on farms where calves were picked up twice daily compared with once daily (18.2 (95% CI=16.5-19.9)% vs. 15.9% (95% CI=15.2-16.6)%; p=0.012), and was lower where first milking colostrum was combined with colostrum obtained at later milkings (15.0 (95% CI=13.9-16.1)%) compared with where it was not (16.9 (95% CI=16.3-17.6)%; p=0.002). Vaccination of all cows against calf diarrhoeal pathogens was associated with increased Brix compared with no vaccination (18.1 (95% CI=16.6-19.6)% vs. 16.3 (95% CI=15.6-17.0)%; p=0.033). Mean pH of samples tested decreased from 5.97 (95% CI=5.84-6.09) to 5.58 (95% CI=5.45-5.71) for early and late-season visits, respectively (p<0.001). Of 268 samples tested, 23 (8.6%) had bacterial counts below the recommended threshold of 1.00×10 5 cfu/mL. Mean bacterial counts increased from 2.75 (95% CI=1.80-3.70)×10 8 to 4.99 (95% CI=3.95-6.03)×10 8 cfu/mL for early and late-season visits, respectively (p<0.001). Of 259 samples tested, 23 (8.9%) had coliform counts below the recommended threshold of 1.00×10 4 cfu/mL. On a large majority of dairy farms included in this study the pooled colostrum fed to newborn calves had sub-optimal Brix and excessive bacterial counts. Farm-level risk factors such as twice daily pick up of calves from the paddocks, herd vaccination and feeding pooled colostrum that did not combine colostrum obtained at later milkings with first milking colostrum were associated with colostrum quality measures.

A substantial fraction of phytoplankton-derived DON is resistant to degradation by a metabolically versatile, widely distributed marine bacterium

PubMed Central

Kimmance, Susan; McCormack, Paul

2017-01-01

The capacity of bacteria for degrading dissolved organic nitrogen (DON) and remineralising ammonium is of importance for marine ecosystems, as nitrogen availability frequently limits productivity. Here, we assess the capacity of a widely distributed and metabolically versatile marine bacterium to degrade phytoplankton-derived dissolved organic carbon (DOC) and nitrogen. To achieve this, we lysed exponentially growing diatoms and used the derived dissolved organic matter (DOM) to support an axenic culture of Alteromonas sp.. Bacterial biomass (as particulate carbon and nitrogen) was monitored for 70 days while growth dynamics (cell count), DOM (DOC, DON) and dissolved nutrient concentrations were monitored for up to 208 days. Bacterial biomass increased rapidly within the first 7 days prior to a period of growth/death cycles potentially linked to rapid nutrient recycling. We found that ≈75% of the initial DOC and ≈35% of the initial DON were consumed by bacteria within 40 and 4 days respectively, leaving a significant fraction of DOM resilient to degradation by this bacterial species. The different rates and extents to which DOC and DON were accessed resulted in changes in DOM stoichiometry and the iterative relationship between DOM quality and bacterial growth over time influenced bacterial cell C:N molar ratio. C:N values increased to 10 during the growth phase before decreasing to values of ≈5, indicating a change from relative N-limitation/C-sufficiency to relative C-limitation/N-sufficiency. Consequently, despite its reported metabolic versatility, we demonstrate that Alteromonas sp. was unable to access all phytoplankton derived DOM and that a bacterial community is likely to be required. By making the relatively simple assumption that an experimentally derived fraction of DOM remains resilient to bacterial degradation, these experimental results were corroborated by numerical simulations using a previously published model describing the interaction between DOM and bacteria in marine systems, thus supporting our hypothesis. PMID:28158278

A substantial fraction of phytoplankton-derived DON is resistant to degradation by a metabolically versatile, widely distributed marine bacterium.

PubMed

Polimene, Luca; Clark, Darren; Kimmance, Susan; McCormack, Paul

2017-01-01

The capacity of bacteria for degrading dissolved organic nitrogen (DON) and remineralising ammonium is of importance for marine ecosystems, as nitrogen availability frequently limits productivity. Here, we assess the capacity of a widely distributed and metabolically versatile marine bacterium to degrade phytoplankton-derived dissolved organic carbon (DOC) and nitrogen. To achieve this, we lysed exponentially growing diatoms and used the derived dissolved organic matter (DOM) to support an axenic culture of Alteromonas sp.. Bacterial biomass (as particulate carbon and nitrogen) was monitored for 70 days while growth dynamics (cell count), DOM (DOC, DON) and dissolved nutrient concentrations were monitored for up to 208 days. Bacterial biomass increased rapidly within the first 7 days prior to a period of growth/death cycles potentially linked to rapid nutrient recycling. We found that ≈75% of the initial DOC and ≈35% of the initial DON were consumed by bacteria within 40 and 4 days respectively, leaving a significant fraction of DOM resilient to degradation by this bacterial species. The different rates and extents to which DOC and DON were accessed resulted in changes in DOM stoichiometry and the iterative relationship between DOM quality and bacterial growth over time influenced bacterial cell C:N molar ratio. C:N values increased to 10 during the growth phase before decreasing to values of ≈5, indicating a change from relative N-limitation/C-sufficiency to relative C-limitation/N-sufficiency. Consequently, despite its reported metabolic versatility, we demonstrate that Alteromonas sp. was unable to access all phytoplankton derived DOM and that a bacterial community is likely to be required. By making the relatively simple assumption that an experimentally derived fraction of DOM remains resilient to bacterial degradation, these experimental results were corroborated by numerical simulations using a previously published model describing the interaction between DOM and bacteria in marine systems, thus supporting our hypothesis.

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.

Comparisons of Internal Behavior after Exposure to Flavobacterium psychrophilum between Two Ayu (Plecoglossus altivelis altivelis) Strains Showing Different Cumulative Mortality to Bacterial Cold Water Disease

PubMed Central

KAGEYAMA, Tetsushi; KUWADA, Tomonori; OHARA, Kenichi; NOUNO, Aya; UMINO, Tetsuya; FURUSAWA, Shuichi

2013-01-01

ABSTRACT Bacterial cold water disease (BCWD) in ayu (Plecoglossus altivelis altivelis) has a serious impact on aquaculture and fisheries. There is known to be a significant difference among ayu strains with regard to mortality caused by BCWD. In this study, the immune response of different ayu strains against Flavobacterium psychrophilum infection was observed. One strain was resistant to infection by F. psychrophilum, and the other was susceptible to infection by the same bacteria. The number of bacteria in the body was observed in each ayu strain, and the change in bacterial counts was similar. However, there was a significant difference in bacterial count in the spleen between the two strains on days 6, 9, 12 and 15 after exposure. To observe the immune response against F. psychrophilum, agglutination assay using serum was performed. An agglutination reaction in the resistant ayu strain was observed in 4 out of 6 ayu on day 6 after exposure, while no reactions in the susceptible ayu strain were observed in any sampled fish until day 12. However, some reactions in the susceptible ayu strain were observed in surviving ayu. These results indicate that there is a correlation between the presence of bacterial multiplication and agglutination reaction against F. psychrophilum. PMID:23902927

Silkworm larvae plasma (SLP) assay for detection of bacteria: False positives secondary to inflammation in vivo.

PubMed

Ma, Michelle; Rice, Tyler A; Percopo, Caroline M; Rosenberg, Helene F

2017-01-01

The silkworm larvae plasma (SLP) assay has been developed as a means to detect bacterial peptidoglycan as a surrogate for live bacteria. Here, we present results that indicate that generation of melanin by this assay is not fully reliable as a surrogate marker for bacterial count. Published by Elsevier B.V.

Establishment of HPC(R2A) for regrowth control in non-chlorinated distribution systems.

PubMed

Uhl, Wolfgang; Schaule, Gabriela

2004-05-01

Drinking water distributed without disinfection and without regrowth problems for many years may show bacterial regrowth when the residence time and/or temperature in the distribution system increases or when substrate and/or bacterial concentration in the treated water increases. An example of a regrowth event in a major German city is discussed. Regrowth of HPC bacteria occurred unexpectedly at the end of a very hot summer. No pathogenic or potentially pathogenic bacteria were identified. Increased residence times in the distribution system and temperatures up to 25 degrees C were identified as most probable causes and the regrowth event was successfully overcome by changing flow regimes and decreasing residence times. Standard plate counts of HPC bacteria using the spread plate technique on nutrient rich agar according to German Drinking Water Regulations (GDWR) had proven to be a very good indicator of hygienically safe drinking water and to demonstrate the effectiveness of water treatment. However, the method proved insensitive for early regrowth detection. Regrowth experiments in the lab and sampling of the distribution system during two summers showed that spread plate counts on nutrient-poor R2A agar after 7-day incubation yielded 100 to 200 times higher counts. Counts on R2A after 3-day incubation were three times less than after 7 days. As the precision of plate count methods is very poor for counts less than 10 cfu/plate, a method yielding higher counts is better suited to detect upcoming regrowth than a method yielding low counts. It is shown that for the identification of regrowth events HPC(R2A) gives a further margin of about 2 weeks for reaction before HPC(GDWR). Copyright 2003 Elsevier B.V.

Comparison of bacterial counts obtained from naturally contaminated foods by means of Stomacher and blender.

PubMed

Purvis, U; Sharpe, A N; Bergener, D M; Lachapelle, G; Milling, M; Spiring, F

1987-01-01

Four Regional Health Protection Branch laboratories each compared aerobic colony counts obtained after "stomaching" and blending, for a minimum of 10 samples in each of the seven food groups: dry pastas; chocolate and cocoa powders; frozen entrees (macaroni and cheese, chow mein, chop suey, fried rice, seafood casseroles, and Salisbury steak); nonfat dry milk; shrimp and crabmeats; spices; and breakfast sausages. Overall, counts obtained after using the Stomacher were equivalent to or higher than counts obtained after using the blender in 73% of the comparisons (alpha = 0.05). Where differences existed, counts obtained after using the Stomacher tended to be higher than counts obtained after using the blender from milk powder and lower from sausage. Aerobic colony counts from these foods are not unacceptably biased when obtained by Stomacher.

Utilization of buffered vinegar to increase the shelf life of chicken retail cuts packaged in carbon dioxide.

PubMed

Desai, Monil A; Kurve, Vikram; Smith, Brian S; Campano, Stephen G; Soni, Kamlesh; Schilling, M Wes

2014-07-01

Poultry processors commonly place whole parts of broilers in plastic packages and seal them in an atmosphere of 100% carbon dioxide before shipping them to food service and retail customers. This practice extends the shelf life of retail cuts to approximately 12 d under refrigerated conditions. The objective of this study was to determine the antimicrobial efficacy of vinegar for growth inhibition of mesophilic and lactic acid bacterial counts and enhancement of shelf life in CO2-packaged refrigerated chicken thigh samples. Meat quality, sensory differences, and microbial enumeration were evaluated for chicken thighs that were sprayed with 0, 0.5, or 1.0% vinegar. No differences were observed (P > 0.05) among treatments (control vs. 0.5 and 1.0% vinegar-treated chicken thighs) with respect to pH and Commission Internationale d'Eclairage L*a*b*for both chicken skin and the meat tissue. The difference from the control test indicated that trained panelists were not able to detect a difference (P > 0.05) in flavor between the chicken thigh treatments. The mesophilic and Lactobacillus bacterial counts were enumerated after 0, 4, 8, 12, 16, and 20 d of storage. The mesophilic bacterial load for the 1.0% vinegar treatment was less than all other treatments after 8, 12, 16, and 20 d of storage, whereas the 0.5% vinegar treatment had lower bacterial counts at d 12 than both controls and had an approximate shelf life of 16 d. For lactic acid bacteria, the vinegar 1.0% treatment had lower counts than the control treatments at d 12 and 16. The results from the study indicate that a combination of 1.0% vinegar with CO2 packaging can extend the shelf life from 12 to 20 d for chicken retail cuts without negatively affecting the quality and sensory properties of the broiler meat. © 2014 Poultry Science Association Inc.

Acute bacterial meningitis cases diagnosed by culture and PCR in a children's hospital throughout a 9-Year period (2000-2008) in Athens, Greece.

PubMed

Papavasileiou, Konstantina; Papavasileiou, Eleni; Tzanakaki, Georgina; Voyatzi, Aliki; Kremastinou, Jenny; Chatzipanagiotou, Stylianos

2011-04-01

Acute bacterial meningitis is one of the most severe infectious diseases, affecting mainly infants and, secondarily, older children and adolescents. Diagnosis in the early stages is often difficult and despite treatment with appropriate antibiotic therapy, the case fatality rate remains high. In the present study, the incidence of bacterial meningitis was registered in a general pediatric hospital in Athens, Greece, during a 9-year period (2000-2008), and the use of molecular methods in the diagnosis of bacterial meningitis versus the conventional cultural methods was evaluated. The impact of vaccination against meningitis-causing bacteria on the incidence of bacterial meningitis was also assessed. From a total of 1833 children hospitalized with suspected clinical symptoms and signs of meningitis, all cerebrospinal fluid (CSF) and blood samples were analyzed by white blood cell (WBC) count, measurement of glucose, protein, and C-reactive protein (CRP) levels, as well as by conventional bacteriologic culture methods. If samples showed altered CSF markers that were consistent with meningitis in general, they were further investigated by PCR for bacterial pathogens. Of the 1833 patients, 289 (15.76%) were found to be positive for meningitis after CSF examination, based on white blood cell count and differentiation, glucose, protein, and CRP. Fifty-six of the 289 (19.37%) had confirmed bacterial meningitis, as diagnosed by either culture and/or PCR. Of these 56 cases, 44 (78.6%) were detected only by PCR, and 12 cases (21.4%) were confirmed by PCR and culture. The predominant microorganism was Neisseria meningitidis serogroup B (n = 40; 71.4%), followed by Streptococcus pneumoniae not typed [NT] (n = 7; 12.5%), Streptococcus spp. (n =4; 7.1%), Haemophilus influenzae NT (n = 2; 3.6%), and S. pneumoniae serotype 3, Streptococcus group B, and S. pneumoniae serotype 18C (each n = 1; 1.8%). In Greece, according to data from the National Meningitis Reference Laboratory, vaccination against N. meningitidis serogroup C since 2001 led to a 10-fold decrease in the incidence of meningitis cases, vaccination against S. pneumoniae serotypes included in the heptavalent conjugate vaccine since 2005 led to a 3.4-fold incidence decrease, and vaccination against H. influenzae type b since 1992 led almost to an absence of cases. In the population of the present study, none of the cases were caused by the above-mentioned vaccine pathogens, except for one S. pneumoniae serotype 18C case with no history of past vaccination. The introduction of vaccination against meningitis-causing bacteria has drastically decreased the emergence of the infection. The improved molecular amplification assays proved to be superior to conventional bacteriologic methods and should be introduced into routine diagnosis, as well as the epidemiologic surveillance of bacterial meningitis.

Bacterial quality and safety of packaged fresh leafy vegetables at the retail level in Finland.

PubMed

Nousiainen, L-L; Joutsen, S; Lunden, J; Hänninen, M-L; Fredriksson-Ahomaa, M

2016-09-02

Consumption of packaged fresh leafy vegetables, which are convenient ready-to-eat products, has increased during the last decade. The number of foodborne outbreaks associated with these products has concurrently increased. In our study, (1) label information, (2) O2/CO2 composition, (3) bacterial quality and (4) safety of 100 fresh leafy vegetables at the retail level were studied in Finland during 2013. Bacterial quality was studied using aerobic bacteria (AB) and coliform bacteria (CB) counts, and searching for the presence of Escherichia coli, Listeria and Yersinia. The safety was studied by the presence of Salmonella, ail-positive Yersinia, stx-positive E. coli (STEC) and Listeria monocytogenes using PCR and culturing. Important label information was unavailable on several packages originating from different companies. The packaging date was missing on all packages and the date of durability on 83% of the packages. Storage temperature was declared on 62% of the packages and 73% of the packages contained information about prewashing. The batch/lot number was missing on 29% of the packages. Very low oxygen (O2) (<1%) and elevated carbon dioxide (CO2) (2-22%) concentrations were measured in all packages labelled to contain a protective atmosphere. O2 and CO2 concentrations varied widely in the rest of the packages. AB and CB counts were high in the leafy vegetable samples varying between 6.2 and 10.6 and 4.2-8.3logcfu/g, respectively. In most of the samples, the AB and CB counts exceeded 10(8) and 10(6)cfu/g, respectively. A positive correlation was observed between the AB and CB counts. E. coli was isolated from 15% of the samples and Yersinia from 33%. L. monocytogenes was isolated from two samples and ail-positive Y. enterocolitica in one. Using PCR, STEC was detected in seven samples, and Salmonella and ail-positive Y. enterocolitica in two samples each. The AB and CB mean values of products originating from different companies varied widely. High AB and CB counts and pathogenic bacteria were detected in ready-to-eat products not needing washing before use. Our study shows that the bacterial quality and safety of packaged fresh leafy vegetables is poor and label information on the packages is inadequate. More studies are needed concerning the impact of a protective atmosphere on bacterial growth, and the impact of washing for removing bacteria. Copyright © 2016 Elsevier B.V. All rights reserved.

No lower bacterial adhesion for ceramics compared to other biomaterials: An in vitro analysis.

PubMed

Slullitel, P A; Buttaro, M A; Greco, G; Oñativia, J I; Sánchez, M L; Mc Loughlin, S; García-Ávila, C; Comba, F; Zanotti, G; Piccaluga, F

2018-06-01

Although there is some clinical evidence of ceramic bearings being associated with a lower infection rate after total hip arthroplasty (THA), available data remains controversial since this surface is usually reserved for young, healthy patients. Therefore, we investigated the influence of five commonly used biomaterials on the adhesion potential of four biofilm-producing bacteria usually detected in infected THAs. Ceramic biomaterials exhibit less bacterial adherence than other biomaterials. In this in vitro research, we evaluated the ability of Staphylococcus aureus, Staphylococcus epidermidis ATCC 35984, Escherichia coli ATCC 25922 and Pseudomonas aeruginosa to adhere to the surface of a cobalt-chromium metal head, a fourth-generation ceramic head, a fourth-generation ceramic insert, a highly-crossed linked polyethylene insert and a titanium porous-coated acetabular component. After an initial washing step, bacterial separation from the surface of each specimen was done with a vortex agitator. The colony-forming units were counted to determine the number of viable adherent bacteria. We found no differences on global bacterial adhesion between the different surfaces (p=0.5). E. coli presented the least adherence potential among the analysed pathogens (p<0.001). The combination of E. coli and S. epidermidis generated an antagonist effect over the adherence potential of S. epidermidis individually (58±4% vs. 48±5%; p=0.007). The combination of P. aeruginosa and S. aureus presented a trend to an increased adherence of P. aeruginosa independently, suggesting an agonist effect (71% vs. 62%; p=0.07). Ceramic bearings appeared not to be related to a lower bacterial adhesion than other biomaterials. However, different adhesive potentials among bacteria may play a major role on infection's inception. IV, in vitro study. Copyright © 2018 Elsevier Masson SAS. All rights reserved.

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.

AIRBORNE MICROORGANISMS IN BROILER PROCESSING PLANTS.

PubMed

KOTULA, A W; KINNER, J A

1964-05-01

Concentrations of total aerobic bacteria, molds, yeasts, coliforms, enterococci, and psychrophiles were determined in the air of two poultry processing plants with Andersen samplers and a mobile power supply. Total aerobic bacterial counts were highest in the dressing room, with diminishing numbers in the shackling, eviscerating, and holding rooms, when sampling was carried out during plant operation. The average counts per ft(3) of air in these four rooms were 2,200; 560; 230; and 62, respectively. (Each value is the average of 36 observations.) The number of organisms increased in the shackling and dressing rooms once processing was begun. Average total aerobic bacterial counts increased from 70 to 870 to 3,000 in the shackling room and from 310 to 4,900 to 7,000 in the dressing room when sampling was carried out at 5:00 am (before plant operations), 9:00 am, and 2:00 pm, respectively. (Each value is the mean of 12 observations.) Airborne molds might originate from a source other than the poultry being processed.

Airborne Microorganisms in Broiler Processing Plants

PubMed Central

Kotula, Anthony W.; Kinner, Jack A.

1964-01-01

Concentrations of total aerobic bacteria, molds, yeasts, coliforms, enterococci, and psychrophiles were determined in the air of two poultry processing plants with Andersen samplers and a mobile power supply. Total aerobic bacterial counts were highest in the dressing room, with diminishing numbers in the shackling, eviscerating, and holding rooms, when sampling was carried out during plant operation. The average counts per ft3 of air in these four rooms were 2,200; 560; 230; and 62, respectively. (Each value is the average of 36 observations.) The number of organisms increased in the shackling and dressing rooms once processing was begun. Average total aerobic bacterial counts increased from 70 to 870 to 3,000 in the shackling room and from 310 to 4,900 to 7,000 in the dressing room when sampling was carried out at 5:00 am (before plant operations), 9:00 am, and 2:00 pm, respectively. (Each value is the mean of 12 observations.) Airborne molds might originate from a source other than the poultry being processed. Images FIG. 3 PMID:14170951

A vegan or vegetarian diet substantially alters the human colonic faecal microbiota.

PubMed

Zimmer, J; Lange, B; Frick, J-S; Sauer, H; Zimmermann, K; Schwiertz, A; Rusch, K; Klosterhalfen, S; Enck, P

2012-01-01

Consisting of ≈10(14) microbial cells, the intestinal microbiota represents the largest and the most complex microbial community inhabiting the human body. However, the influence of regular diets on the microbiota is widely unknown. We examined faecal samples of vegetarians (n=144), vegans (n=105) and an equal number of control subjects consuming ordinary omnivorous diet who were matched for age and gender. We used classical bacteriological isolation, identification and enumeration of the main anaerobic and aerobic bacterial genera and computed absolute and relative numbers that were compared between groups. Total counts of Bacteroides spp., Bifidobacterium spp., Escherichia coli and Enterobacteriaceae spp. were significantly lower (P=0.001, P=0.002, P=0.006 and P=0.008, respectively) in vegan samples than in controls, whereas others (E. coli biovars, Klebsiella spp., Enterobacter spp., other Enterobacteriaceae, Enterococcus spp., Lactobacillus spp., Citrobacter spp. and Clostridium spp.) were not. Subjects on a vegetarian diet ranked between vegans and controls. The total microbial count did not differ between the groups. In addition, subjects on a vegan or vegetarian diet showed significantly (P=0.0001) lower stool pH than did controls, and stool pH and counts of E. coli and Enterobacteriaceae were significantly correlated across all subgroups. Maintaining a strict vegan or vegetarian diet results in a significant shift in the microbiota while total cell numbers remain unaltered.

Salivary bacterial leakage into implant-abutment connections: preliminary results of an in vitro study.

PubMed

Mencio, F; Papi, P; Di Carlo, S; Pompa, G

2016-06-01

The occurrence of bacterial leakage in the internal surface of implants, through implant-abutment interface (IAI), is one of the parameters for analyzing the fabrication quality of the connections. The aim of this in vitro study is to evaluate two different types of implant-abutment connections: the screwed connection (Group 1) and the cemented connection (Group 2), analyzing the permeability of the IAI to bacterial colonization, using human saliva as culture medium. A total of twelve implants were tested, six in each experimental group. Five healthy patients were enrolled in this study. Two milliliters of non-stimulated saliva were collected from each subject and mixed in a test tube. After 14 days of incubation of the bacteria sample in the implant fixtures, a PCR-Real Time analysis was performed. Fisher's exact test was used to compare the proportions of implant-abutment assembled structures detected with bacterial leakage. Differences in the bacterial counts of the two groups were compared using the Mann-Whitney U test. A p value < 0.05 was considered significant. The results showed a decreased stability with the screwed implant-abutment connections compared to the cemented implant-abutment connections. A mean total bacterial count of 1.2E+07 (± 0.25E+07) for Group 1 and of 7.2E+04 (± 14.4E+04) for Group 2 was found, with a high level of significance, p = .0001. Within the limitations of this study it can be concluded that bacterial species from human saliva may penetrate along the implant-abutment interface in both connections, however the cemented connection implants showed the lowest amount of bacterial colonization.

Bacterial adhesion on direct and indirect dental restorative composite resins: An in vitro study on a natural biofilm.

PubMed

Derchi, Giacomo; Vano, Michele; Barone, Antonio; Covani, Ugo; Diaspro, Alberto; Salerno, Marco

2017-05-01

Both direct and indirect techniques are used for dental restorations. Which technique should be preferred or whether they are equivalent with respect to bacterial adhesion is unclear. The purpose of this in vitro study was to determine the affinity of bacterial biofilm to dental restorative composite resins placed directly and indirectly. Five direct composite resins for restorations (Venus Diamond, Adonis, Optifil, Enamel Plus HRi, Clearfil Majesty Esthetic) and 3 indirect composite resins (Gradia, Estenia, Signum) were selected. The materials were incubated in unstimulated whole saliva for 1 day. The biofilms grown were collected and their bacterial cells counted. In parallel, the composite resin surface morphology was analyzed with atomic force microscopy. Both bacterial cell count and surface topography parameters were subjected to statistical analysis (α=.05). Indirect composite resins showed significantly lower levels than direct composite resins for bacterial cell adhesion, (P<.001). No significant differences were observed within the direct composite resins (P>.05). However, within the indirect composite resins a significantly lower level was found for Gradia than Estenia or Signum (P<.01). A partial correlation was observed between composite resin roughness and bacterial adhesion when the second and particularly the third-order statistical moments of the composite resin height distributions were considered. Indirect dental restorative composite resins were found to be less prone to biofilm adhesion than direct composite resins. A correlation of bacterial adhesion to surface morphology exists that is described by kurtosis; thus, advanced data analysis is required to discover possible insights into the biologic effects of morphology. Copyright © 2016 Editorial Council for the Journal of Prosthetic Dentistry. Published by Elsevier Inc. All rights reserved.

Plasma surface modification of rigid contact lenses decreases bacterial adhesion.

PubMed

Wang, Yingming; Qian, Xuefeng; Zhang, Xiaofeng; Xia, Wei; Zhong, Lei; Sun, Zhengtai; Xia, Jing

2013-11-01

Contact lens safety is an important topic in clinical studies. Corneal infections usually occur because of the use of bacteria-carrying contact lenses. The current study investigated the impact of plasma surface modification on bacterial adherence to rigid contact lenses made of fluorosilicone acrylate materials. Boston XO and XO2 contact lenses were modified using plasma technology (XO-P and XO2-P groups). Untreated lenses were used as controls. Plasma-treated and control lenses were incubated in solutions containing Staphylococcus aureus or Pseudomonas aeruginosa. MTT colorimetry, colony-forming unit counting method, and scanning electron microscopy were used to measure bacterial adhesion. MTT colorimetry measurements showed that the optical density (OD) values of XO-P and XO2-P were significantly lower than those of XO and XO2, respectively, after incubation with S. aureus (P < 0.01). The OD value of XO-P was also much lower than that of XO after incubation with P. aeruginosa (P < 0.01). Colony-forming unit counting revealed that a significantly lower number of bacterial colonies attached to the XO-P versus XO lenses and to the XO2-P versus XO2 lenses incubated with S. aureus (P < 0.01). Fewer bacterial colonies attached to the XO-P versus XO lenses incubated with P. aeruginosa (P < 0.01). Further, scanning electron microscopy suggested different bacterial adhesion morphology on plasma-treated versus control lenses. Plasma surface modification can significantly decrease bacterial adhesion to fluorosilicone acrylate contact lenses. This study provides important evidence of a unique benefit of plasma technology in contact lens surface modification.

USE OF SCORE AND CEREBROSPINAL FLUID LACTATE DOSAGE IN DIFFERENTIAL DIAGNOSIS OF BACTERIAL AND ASEPTIC MENINGITIS.

PubMed

Pires, Frederico Ribeiro; Franco, Andréia Christine Bonotto Farias; Gilio, Alfredo Elias; Troster, Eduardo Juan

2017-01-01

To evaluate Bacterial Meningitis Score (BMS) on its own and in association with Cerebrospinal Fluid (CSF) lactate dosage in order to distinguish bacterial from aseptic meningitis. Children diagnosed with meningitis at a tertiary hospital between January/2011 and December/2014 were selected. All data were obtained upon admission. BMS was applied and included: CSF Gram staining (2 points); CSF neutrophil count ≥1,000 cells/mm3 (1 point); CSF protein ≥80 mg/dL (1 point); peripheral blood neutrophil count ≥10,000 cells/mm3 (1 point) and seizures upon/before arrival (1 point). Cutoff value for CSF lactate was ≥30 mg/dL. Sensitivity, specificity and negative predictive value of several BMS cutoffs and BMS associated with high CSF lactate were evaluated for prediction of bacterial meningitis. Among 439 eligible patients, 94 did not have all data available to complete the score, and 345 patients were included: 7 in bacterial meningitis group and 338 in aseptic meningitis group. As predictive factors of bacterial meningitis, BMS ≥1 had 100% sensitivity (95%CI 47.3-100), 64.2% specificity (58.8-100) and 100% negative predictive value (97.5-100); BMS ≥2 or BMS ≥1 associated with high CSF lactate also showed 100% sensitivity (47.3-100); but 98.5% specificity (96.6-99.5) and 100% negative predictive value (98.3-100). 2 point BMS in association with CSF lactate dosage had the same sensitivity and negative predictive value, with increased specificity for diagnosis of bacterial meningitis when compared with 1-point BMS.

Performance of a veterinary urine dipstick paddle system for diagnosis and identification of urinary tract infections in dogs and cats.

PubMed

Ybarra, Winnie L; Sykes, Jane E; Wang, Yenlie; Byrne, Barbara A; Westropp, Jodi L

2014-04-01

To evaluate the performance of a veterinary urine dipstick paddle (UDP) for diagnosis and identification of urinary tract infection (UTI) in dogs and cats. Prospective, randomized, blinded study. 207 urine specimens. UDPs were inoculated by 2 investigators and incubated according to manufacturer's instructions. Results, including presence or absence of bacterial growth, organism counts, and identification of uropathogens, were compared between investigators and with microbiology laboratory results. A subset of UDPs with bacterial growth was submitted to the laboratory for confirmation. The laboratory reported 64 (30.9%) specimens had growth of bacteria. Bacterial growth was reported for 63 (30.4%) and 58 (28.0%) of the UDPs by investigators 1 and 2, respectively. Sensitivity and specificity of the UDP for detection of bacterial growth were 97.3% and 98.6%, respectively, for investigator 1 and 89.1% and 99.3%, respectively, for investigator 2. For UPDs with ≥ 10(5) colony-forming units/mL, organism counts correlated well between the laboratory and investigators 1 (r = 0.95) and 2 (r = 0.89). Pathogen identification was not always accurate. Only 25 of 33 (75.8%) UDPs submitted for confirmation yielded bacteria consistent with those isolated from the original bacterial culture of urine. The veterinary UDP system was a sensitive test for screening patients for bacterial UTI, but uropathogen identification was not always accurate. When UDPs have bacterial growth, a fresh urine specimen should be submitted to the laboratory to confirm the identity of the organisms and to permit antimicrobial susceptibility testing.

Clinical Value of Assessing Cytokine Levels for the Differential Diagnosis of Bacterial Meningitis in a Pediatric Population

PubMed Central

Ye, Qing; Shao, Wen-Xia; Shang, Shi-Qiang; Shen, Hong-Qiang; Chen, Xue-Jun; Tang, Yong-Min; Yu, Yong-Lin; Mao, Jian-Hua

2016-01-01

Abstract We performed a prospective observational study to evaluate the utility of measuring inflammatory cytokine levels to discriminate bacterial meningitis from similar common pediatric diseases. Inflammatory cytokine levels and other cerebrospinal fluid (CSF) physicochemical indicators were evaluated in 140 patients who were diagnosed with bacterial meningitis via microbiological culture or PCR assay. The CSF concentrations of interleukin (IL)-6 and IL-10, CSF/blood IL-6 and IL-10 ratios, CSF white blood cell count, and CSF micro total protein were significantly elevated in bacterial meningitis patients compared with healthy children or patients with viral encephalitis, epilepsy, or febrile convulsions (P < 0.001). The area under the curve values for CSF concentrations of IL-6 and IL-10, CSF/blood IL-6 and IL-10 ratios, CSF white blood cell count, and CSF micro total protein to identify bacterial meningitis episodes by receiver-operating characteristic analysis were 0.988, 0.949, 0.995, 0.924, 0.945, and 0.928, respectively. The area under the curve for the combination of CSF IL-6 and CSF/blood IL-6 ratio was larger than that for either parameter alone, and the combination exhibited enhanced specificity and positive predictive value. After effective meningitis treatment, CSF IL-6 levels dropped significantly. These results suggest that CSF IL-6 and CSF/blood IL-6 ratio are good biomarkers in discriminating bacterial meningitis. Evaluating CSF IL-6 and CSF/blood IL-6 ratio in combination can improve diagnostic efficiency. Additionally, CSF IL-6 levels can be used to monitor the effects of bacterial meningitis treatment. PMID:27043692

Clinical Value of Assessing Cytokine Levels for the Differential Diagnosis of Bacterial Meningitis in a Pediatric Population.

PubMed

Ye, Qing; Shao, Wen-Xia; Shang, Shi-Qiang; Shen, Hong-Qiang; Chen, Xue-Jun; Tang, Yong-Min; Yu, Yong-Lin; Mao, Jian-Hua

2016-03-01

We performed a prospective observational study to evaluate the utility of measuring inflammatory cytokine levels to discriminate bacterial meningitis from similar common pediatric diseases. Inflammatory cytokine levels and other cerebrospinal fluid (CSF) physicochemical indicators were evaluated in 140 patients who were diagnosed with bacterial meningitis via microbiological culture or PCR assay. The CSF concentrations of interleukin (IL)-6 and IL-10, CSF/blood IL-6 and IL-10 ratios, CSF white blood cell count, and CSF micro total protein were significantly elevated in bacterial meningitis patients compared with healthy children or patients with viral encephalitis, epilepsy, or febrile convulsions (P < 0.001). The area under the curve values for CSF concentrations of IL-6 and IL-10, CSF/blood IL-6 and IL-10 ratios, CSF white blood cell count, and CSF micro total protein to identify bacterial meningitis episodes by receiver-operating characteristic analysis were 0.988, 0.949, 0.995, 0.924, 0.945, and 0.928, respectively. The area under the curve for the combination of CSF IL-6 and CSF/blood IL-6 ratio was larger than that for either parameter alone, and the combination exhibited enhanced specificity and positive predictive value. After effective meningitis treatment, CSF IL-6 levels dropped significantly. These results suggest that CSF IL-6 and CSF/blood IL-6 ratio are good biomarkers in discriminating bacterial meningitis. Evaluating CSF IL-6 and CSF/blood IL-6 ratio in combination can improve diagnostic efficiency. Additionally, CSF IL-6 levels can be used to monitor the effects of bacterial meningitis treatment.

Comparison of bacterial counts in expressed breast milk following standard or strict infection control regimens in neonatal intensive care units: compliance of mothers does matter.

PubMed

Haiden, N; Pimpel, B; Assadian, O; Binder, C; Kreissl, A; Repa, A; Thanhäuser, M; Roberts, C D; Berger, A

2016-03-01

Bacterial counts in 1466 expressed breast milk (EBM) samples from women following one of two infection control regimens (standard vs strict) were investigated. Overall, 12% of samples yielded Gram-negative bacteria, with no significant differences between the standard [11.9% (94/788)] and strict [12.1% (82/678)] regimens (P = 0.92). Significantly more samples were contaminated when expressed at home (standard regimen home/hospital: 17.9% vs 6.1%; strict regimen home/hospital: 19.6% vs 3.4%; P < 0.001). Bacterial contamination of EBM was not associated with the regimen, but was associated with the location of breast milk expression. Attempts to improve personal hygiene during milk collection seem to be of limited value. Good hygiene of collection and storage equipment is likely to be the most important way to ensure the microbiological quality of EBM. Copyright © 2016 The Healthcare Infection Society. Published by Elsevier Ltd. All rights reserved.

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.

Efficacy of contact lens disinfecting solutions against Staphylococcus aureus and Pseudomonas aeruginosa.

PubMed

Manuj, Kapur; Gunderson, Charlise; Troupe, John; Huber, Mary E

2006-07-01

To evaluate the disinfection properties of multipurpose contact lens disinfection solutions, based on the International Organization for Standardization (ISO) 14729 guidelines. ReNu with MoistureLoc Multi-Purpose Solution, OPTI-FREE Express with Aldox Multi-Purpose Solution, Betadine 5% sterile ophthalmic preparation solution (povidone iodine), and 0.9% normal saline solution were inoculated with strains of Staphylococcus aureus and Pseudomonas aeruginosa. Surviving bacteria were quantified at specified times. ReNu with MoistureLoc, OPTI-FREE Express, and 5% ophthalmic povidone iodine were effective in achieving a 5-log reduction in bacterial count. Additionally, all three products maintained their effectivity at 72 hours. However, ReNu with MoistureLoc and povidone iodine resulted in the greatest reduction in bacterial colonization. ReNu with MoistureLoc, OPTI-FREE Express, and 5% ophthalmic povidone iodine meet the ISO 14729 guidelines for standalone contact lens solutions. However, ReNu with MoistureLoc and 5% ophthalmic povidone iodine are most efficient in reducing and maintaining low bacterial count for a period of 72 hours.

Fluorometry as a bacterial source tracking tool in coastal watersheds, Trinidad, CA

Treesearch

Trever Parker; Andrew Stubblefield

2012-01-01

Bacterial counts have long been used as indicators of water pollution that may affect public health. By themselves, bacteria are indicators only and can not be used to identify the source of the pollutant for remediation efforts. Methods of microbial source tracking are generally time consuming, labor intensive and expensive. As an alternative, a fluorometer can be...

Low Circulating Natural Killer Cell Counts are Associated With Severe Disease in Patients With Common Variable Immunodeficiency.

PubMed

Ebbo, Mikael; Gérard, Laurence; Carpentier, Sabrina; Vély, Frédéric; Cypowyj, Sophie; Farnarier, Catherine; Vince, Nicolas; Malphettes, Marion; Fieschi, Claire; Oksenhendler, Eric; Schleinitz, Nicolas; Vivier, Eric

2016-04-01

Natural Killer (NK) cells have been shown to exert antiviral and antitumoural activities. Nevertheless most available data are derived from mouse models and functions of these cells in human remain unclear. To evaluate the impact of low circulating NK cell counts and to provide some clues to the role of NK cells in natural conditions, we studied a large cohort of patients with common variable immunodeficiency (CVID) included in a multicenter cohort of patients with primary hypogammaglobulinaemia. Patients were classified into three groups on the basis of their NK cell counts: severe and mild NK cell lymphopenia (<50 and 50-99×10(6)/L respectively), and normal NK cell counts (>100×10(6)/L). Clinical events were analyzed and compared between these three groups of patients. During study period, 457 CVID patients were included: 99 (21.7%) with severe NK cell lymphopenia, 118 (25.8%) with mild NK cell lymphopenia and 240 (52.5%) with normal NK cell counts. Non-infectious complications (57% vs. 36% and 35%), and, particularly, granulomatous complications (25.3% vs. 13.6% and 8.8%), were more frequent in patients with severe NK cell lymphopenia than in other groups. Invasive infections (68.7% vs. 60.2% and 48.8%), including bacteraemia (22.2% vs. 5.9% and 8.3%) and infectious pneumonia (63.6% vs. 59.3% and 44.2%), were also more frequent in this population. However, no difference was observed for viral infections and neoplasms. Low circulating NK cell counts are associated with more severe phenotypes of CVID, which may indicate a protective role of these immune cells against severe bacterial infections and other complications and non-redundant immune functions when the adaptive immune response is not optimal. Copyright © 2016 The Authors. Published by Elsevier B.V. All rights reserved.

Comparison of epifluorescent viable bacterial count methods

NASA Technical Reports Server (NTRS)

Rodgers, E. B.; Huff, T. L.

1992-01-01

Two methods, the 2-(4-Iodophenyl) 3-(4-nitrophenyl) 5-phenyltetrazolium chloride (INT) method and the direct viable count (DVC), were tested and compared for their efficiency for the determination of the viability of bacterial populations. Use of the INT method results in the formation of a dark spot within each respiring cell. The DVC method results in elongation or swelling of growing cells that are rendered incapable of cell division. Although both methods are subjective and can result in false positive results, the DVC method is best suited to analysis of waters in which the number of different types of organisms present in the same sample is assumed to be small, such as processed waters. The advantages and disadvantages of each method are discussed.

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

Changes in bacterial and archaeal communities during the concentration of brine at the graduation towers in Ciechocinek spa (Poland).

PubMed

Kalwasińska, Agnieszka; Deja-Sikora, Edyta; Burkowska-But, Aleksandra; Szabó, Attila; Felföldi, Támas; Kosobucki, Przemysław; Krawiec, Arkadiusz; Walczak, Maciej

2018-03-01

This study evaluates the changes in bacterial and archaeal community structure during the gradual evaporation of water from the brine (extracted from subsurface Jurassic deposits) in the system of graduation towers located in Ciechocinek spa, Poland. The communities were assessed with 16S rRNA gene sequencing (MiSeq, Illumina) and microscopic methods. The microbial cell density determined by direct cell count was at the order of magnitude of 10 7 cells/mL. It was found that increasing salt concentration was positively correlated with both the cell counts, and species-level diversity of bacterial and archaeal communities. The archaeal community was mostly constituted by members of the phylum Euryarchaeota, class Halobacteria and was dominated by Halorubrum-related sequences. The bacterial community was more diverse, with representatives of the phyla Proteobacteria and Bacteroidetes as the most abundant. The proportion of Proteobacteria decreased with increasing salt concentration, while the proportion of Bacteroidetes increased significantly in the more concentrated samples. Representatives of the genera Idiomarina, Psychroflexus, Roseovarius, and Marinobacter appeared to be tolerant to changes of salinity. During the brine concentration, the relative abundances of Sphingobium and Sphingomonas were significantly decreased and the raised contributions of genera Fabibacter and Fodinibius were observed. The high proportion of novel (not identified at 97% similarity level) bacterial reads (up to 42%) in the 16S rRNA gene sequences indicated that potentially new bacterial taxa inhabit this unique environment.

Effect of levofloxacin treatment on semen hyperviscosity in chronic bacterial prostatitis patients.

PubMed

Vicari, L O; Castiglione, R; Salemi, M; Vicari, B O; Mazzarino, M C; Vicari, E

2016-05-01

Changes in seminal fluid viscosity (SFV), reactive oxygen species (ROS) production, cytokines and seminal leucocyte concentration related to microbiological outcome in patients with chronic bacterial prostatitis (CBP) were studied. One hundred and ten infertile patients with CBP (positive sperm culture ≥10(5) colony-forming units [CFU] ml(-1), pathogens or Chlamydia in expressed prostatic secretions) were treated with levofloxacin 500 mg daily for 14 consecutive days per month for 3 months. In case of bacterial prostatitis, two conditions were examined: responders, eradication of 0 to <10(3) CFU ml(-1) (n = 78) and poor responders, >10(3) to <10(5) CFU ml(-1) (n = 32). Compared with poor responders, responders showed a significant increase of sperm progressive motility and a significant decrease in seminal leucocyte count, SFV, liquefaction time, ROS production (in all fractions and conditions), seminal tumour necrosis factor-α and interleukin 6. None of these variables showed significant differences compared with a control group of 37 fertile men. On the other hand, the poor responders showed significant changes in these variables compared with matched pretreatment values. In patients with CBP, antibiotic therapy alone leads to eradication in ≈71%, with improvement of sperm progressive motility, SFV and the framework of prooxidative factors. However, in the remaining ≈29% with poor antibiotic responsiveness, a deterioration of all variables is observed. © 2015 Blackwell Verlag GmbH.

In situ production of human β defensin-3 in lager yeasts provides bactericidal activity against beer-spoiling bacteria under fermentation conditions.

PubMed

James, T C; Gallagher, L; Titze, J; Bourke, P; Kavanagh, J; Arendt, E; Bond, U

2014-02-01

To examine the use of a natural antimicrobial peptide, human β-defensin-3 (HBD3), as a means of preventing spoilage from bacterial contamination in brewery fermentations and in bottled beer. A chemically synthesised HBD3 peptide was tested for bactericidal activity against common Gram-positive and Gram-negative beer-spoiling bacteria, including species of Lactobacillus, Pediococcus and Pectinatus. The peptide was effective at the μmol l(-1) range in vitro, reducing bacterial counts by 95%. A gene construct encoding a secretable form of HBD3 was integrated into the genome of the lager yeast Saccharomyces pastorianus strain CMBS-33. The integrated gene was expressed under fermentation conditions and was secreted from the cell into the medium, but a significant amount remains associated with yeast cell surface. We demonstrate that under pilot-scale fermentation conditions, secreted HBD3 possesses bactericidal activity against beer-spoiling bacteria. Furthermore, when added to bottled beer, a synthetic form of HBD3 reduces the growth of beer-spoiling bacteria. Defensins provide prophylactic protection against beer-spoiling bacteria under brewing conditions and also in bottled beer. The results have direct application to the brewing industry where beer spoilage due to bacterial contamination continues to be a major problem in breweries around the world. © 2013 The Society for Applied Microbiology.

Measurement of lipocalin-2 and syndecan-4 levels to differentiate bacterial from viral infection in children with community-acquired pneumonia.

PubMed

Esposito, Susanna; Bianchini, Sonia; Gambino, Monia; Madini, Barbara; Di Pietro, Giada; Umbrello, Giulia; Presicce, Maria Lory; Ruggiero, Luca; Terranova, Leonardo; Principi, Nicola

2016-07-20

In this study, we evaluated the lipocalin-2 (LIP2) and syndecan-4 (SYN4) levels in children who were hospitalized for radiologically confirmed CAP in order to differentiate bacterial from viral infection. The results regarding the LIP2 and SYN4 diagnostic outcomes were compared with the white blood cell (WBC) count and C reactive protein (CRP) levels. A total of 110 children <14 years old who were hospitalized for radiologically confirmed CAP were enrolled. Serum samples were obtained upon admission and on day 5 to measure the levels of LIP2, SYN4, and CRP as well as the WBC. Polymerase chain reaction of the respiratory secretions and tests on blood samples were performed to detect respiratory viruses, Streptococcus pneumoniae, and Mycoplasma pneumoniae. CAP was considered to be due to a probable bacterial infection in 74 children (67.3 %) and due to a probable viral infection in 16 children (14.5 %). Overall, 84 children (76.4 %) were diagnosed with severe CAP. The mean values of the WBC count and the LIP2 and SYN4 levels did not differ among the probable bacterial, probable viral, and undetermined cases. However, the CRP serum concentrations were significantly higher in children with probable bacterial CAP than in those with probable viral disease (32.2 ± 55.5 mg/L vs 9.4 ± 17.0 mg/L, p < 0.05). The WBC count was the best predictor of severe CAP, but the differences among the studied variables were marginal. The WBC count was significantly lower on day 5 in children with probable bacterial CAP (p < 0.01) and in those with an undetermined etiology (p < 0.01). The CRP and LIP2 levels were significantly lower 5 days after enrollment in all of the studied groups, independent of the supposed etiology of CAP (p < 0.01 for all comparisons). No statistically significant variation was observed for SYN4. Measuring the LIP2 and SYN4 levels does not appear to solve the problem of the poor reliability of routine laboratory tests in defining the etiology and severity of pediatric CAP. Currently, the CRP levels and WBC, when combined with evaluation of clinical data, can be used to limit the overuse of antibiotics as much as possible and to provide the best treatment to the patient.

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

PubMed

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

2018-04-01

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

A Characterization of the Oral Microbiome in Allogeneic Stem Cell Transplant Patients

PubMed Central

Ames, Nancy J.; Sulima, Pawel; Ngo, Thoi; Barb, Jennifer; Munson, Peter J.; Paster, Bruce J.; Hart, Thomas C.

2012-01-01

Background The mouth is a complex biological structure inhabited by diverse bacterial communities. The purpose of this study is to describe the effects of allogeneic stem cell transplantation on the oral microbiota and to examine differences among those patients who acquired respiratory complications after transplantation. Methodology/Principal Findings All patients were consented at the National Institutes of Health, Clinical Center. Bacterial DNA was analyzed from patients' oral specimens using the Human Oral Microbe Identification Microarray. The specimens were collected from four oral sites in 45 allogeneic transplantation patients. Specimens were collected at baseline prior to transplantation, after transplantation at the nadir of the neutrophil count and after myeloid engraftment. If respiratory signs and symptoms developed, additional specimens were obtained. Patients were followed for 100 days post transplantation. Eleven patients' specimens were subjected to further statistical analysis. Many common bacterial genera, such as Streptococcus, Veillonella, Gemella, Granulicatella and Camplyobacter were identified as being present before and after transplantation. Five of 11 patients developed respiratory complications following transplantation and there was preliminary evidence that the oral microbiome changed in their oral specimens. Cluster analysis and principal component analysis revealed this change in the oral microbiota. Conclusions/Significance After allogeneic transplantation, the oral bacterial community's response to a new immune system was not apparent and many of the most common core oral taxa remained unaffected. However, the oral microbiome was affected in patients who developed respiratory signs and symptoms after transplantation. The association related to the change in the oral microbiota and respiratory complications after transplantation will be validated by future studies using high throughput molecular methods. PMID:23144704

Hyperglycemia impedes lung bacterial clearance in a murine model of cystic fibrosis-related diabetes

PubMed Central

Hunt, William R.; Zughaier, Susu M.; Guentert, Dana E.; Shenep, Melissa A.; Koval, Michael; McCarty, Nael A.

2013-01-01

Cystic fibrosis-related diabetes (CFRD) is the most common comorbidity associated with cystic fibrosis (CF), impacting more than half of patients over age 30. CFRD is clinically significant, portending accelerated decline in lung function, more frequent pulmonary exacerbations, and increased mortality. Despite the profound morbidity associated with CFRD, little is known about the underlying CFRD-related pulmonary pathology. Our aim was to develop a murine model of CFRD to explore the hypothesis that elevated glucose in CFRD is associated with reduced lung bacterial clearance. A diabetic phenotype was induced in gut-corrected CF transmembrane conductance regulator (CFTR) knockout mice (CFKO) and their CFTR-expressing wild-type littermates (WT) utilizing streptozotocin. Mice were subsequently challenged with an intratracheal inoculation of Pseudomonas aeruginosa (PAO1) (75 μl of 1–5 × 106 cfu/ml) for 18 h. Bronchoalveolar lavage fluid was collected for glucose concentration and cell counts. A portion of the lung was homogenized and cultured as a measure of the remaining viable PAO1 inoculum. Diabetic mice had increased airway glucose compared with nondiabetic mice. The ability to clear bacteria from the lung was significantly reduced in diabetic WT mice and control CFKO mice. Critically, bacterial clearance by diabetic CFKO mice was significantly more diminished compared with nondiabetic CFKO mice, despite an even more robust recruitment of neutrophils to the airways. This finding that CFRD mice boast an exaggerated, but less effective, inflammatory cell response to intratracheal PAO1 challenge presents a novel and useful murine model to help identify therapeutic strategies that promote bacterial clearance in CFRD. PMID:24097557

Comparison of irrigation solutions and devices in a contaminated musculoskeletal wound survival model.

PubMed

Owens, Brett D; White, Daniel W; Wenke, Joseph C

2009-01-01

There is much to learn about the effectiveness of different methods currently used for the irrigation of open wounds. The purpose of this study was to compare various approaches in a survival animal model. We used an established goat model involving the creation of a reproducible complex musculoskeletal wound followed by inoculation with Pseudomonas aeruginosa (lux) bacteria. This genetically altered luminescent bacterium provides the ability for quantitative analysis with a photon-counting camera system. For Study 1, wound irrigation was performed six hours after the injury and inoculation; the goats were assigned to four treatment groups: normal saline solution, bacitracin solution, castile soap, and benzalkonium chloride. All wounds received sharp débridement and irrigation with use of a pulsatile lavage device (19 psi). Images and photon counts were obtained prior to irrigation, after irrigation, and forty-eight hours after injury and inoculation. For Study 2, we used the same animal model and compared bulb syringe and pulsatile lavage irrigation with saline solution. In Study 1, the irrigation treatment lowered the bacterial counts in all treatment groups. The greatest reduction was seen with castile soap, which lowered the photon count to 13% of the pretreatment level. This was followed by benzalkonium chloride, bacitracin, and saline solution at 18%, 22%, and 29%, respectively. At forty-eight hours, imaging showed a rebound in bacterial counts in every group. The highest rebound was measured in the castile soap group, which rebounded to 120% of the pretreatment level. The benzalkonium chloride group experienced a rebound to 94% of the pretreatment level. These were followed by bacitracin solution (89%) and normal saline solution (68%). In Study 2, both treatment methods were effective in removing 75% of the bacteria initially. At forty-eight hours, the bacterial levels in the pulsed lavage group rebounded to 94% of the original levels (compared with 48% in the bulb syringe group). The difference in the mean photon count ratios at forty-eight hours was significant (p = 0.048). Approaches used to remove bacteria from wounds, such as irrigants other than saline solution or high-pressure devices, may not have the best clinical outcome.

Real-time monitoring of bacterial infection in vivo: development of bioluminescent staphylococcal foreign-body and deep-thigh-wound mouse infection models.

PubMed

Kuklin, Nelly A; Pancari, Gregory D; Tobery, Timothy W; Cope, Leslie; Jackson, Jesse; Gill, Charles; Overbye, Karen; Francis, Kevin P; Yu, Jun; Montgomery, Donna; Anderson, Annaliesa S; McClements, William; Jansen, Kathrin U

2003-09-01

Staphylococcal infections associated with catheter and prosthetic implants are difficult to eradicate and often lead to chronic infections. Development of novel antibacterial therapies requires simple, reliable, and relevant models for infection. Using bioluminescent Staphylococcus aureus, we have adapted the existing foreign-body and deep-wound mouse models of staphylococcal infection to allow real-time monitoring of the bacterial colonization of catheters or tissues. This approach also enables kinetic measurements of bacterial growth and clearance in each infected animal. Persistence of infection was observed throughout the course of the study until termination of the experiment at day 16 in a deep-wound model and day 21 in the foreign-body model, providing sufficient time to test the effects of antibacterial compounds. The usefulness of both animal models was assessed by using linezolid as a test compound and comparing bioluminescent measurements to bacterial counts. In the foreign-body model, a three-dose antibiotic regimen (2, 5, and 24 h after infection) resulted in a decrease in both luminescence and bacterial counts recovered from the implant compared to those of the mock-treated infected mice. In addition, linezolid treatment prevented the formation of subcutaneous abscesses, although it did not completely resolve the infection. In the thigh model, the same treatment regimen resulted in complete resolution of the luminescent signal, which correlated with clearance of the bacteria from the thighs.

Effect of mixing techniques on bacterial attachment and disinfection time of polyether impression material

PubMed Central

Guler, Umut; Budak, Yasemin; Ruh, Emrah; Ocal, Yesim; Canay, Senay; Akyon, Yakut

2013-01-01

Objective: The aim of this study was 2-fold. The first aim was to evaluate the effects of mixing technique (hand-mixing or auto-mixing) on bacterial attachment to polyether impression materials. The second aim was to determine whether bacterial attachment to these materials was affected by length of exposure to disinfection solutions. Materials and Methods: Polyether impression material samples (n = 144) were prepared by hand-mixing or auto-mixing. Escherichia coli, Staphylococcus aureus and Pseudomonas aeruginosa were used in testing. After incubation, the bacterial colonies were counted and then disinfectant solution was applied. The effect of disinfection solution was evaluated just after the polymerization of impression material and 30 min after polymerization. Differences in adherence of bacteria to the samples prepared by hand-mixing and to those prepared by auto-mixing were assessed by Kruskal-Wallis and Mann-Whitney U-tests. For evaluating the efficiency of the disinfectant, Kruskal-Wallis multiple comparisons test was used. Results: E. coli counts were higher in hand-mixed materials (P < 0.05); no other statistically significant differences were found between hand- and auto-mixed materials. According to the Kruskal-Wallis test, significant differences were found between the disinfection procedures (Z > 2.394). Conclusion: The methods used for mixing polyether impression material did not affect bacterial attachment to impression surfaces. In contrast, the disinfection procedure greatly affects decontamination of the impression surface. PMID:24966729

Effect of mixing techniques on bacterial attachment and disinfection time of polyether impression material.

PubMed

Guler, Umut; Budak, Yasemin; Ruh, Emrah; Ocal, Yesim; Canay, Senay; Akyon, Yakut

2013-09-01

The aim of this study was 2-fold. The first aim was to evaluate the effects of mixing technique (hand-mixing or auto-mixing) on bacterial attachment to polyether impression materials. The second aim was to determine whether bacterial attachment to these materials was affected by length of exposure to disinfection solutions. Polyether impression material samples (n = 144) were prepared by hand-mixing or auto-mixing. Escherichia coli, Staphylococcus aureus and Pseudomonas aeruginosa were used in testing. After incubation, the bacterial colonies were counted and then disinfectant solution was applied. The effect of disinfection solution was evaluated just after the polymerization of impression material and 30 min after polymerization. Differences in adherence of bacteria to the samples prepared by hand-mixing and to those prepared by auto-mixing were assessed by Kruskal-Wallis and Mann-Whitney U-tests. For evaluating the efficiency of the disinfectant, Kruskal-Wallis multiple comparisons test was used. E. coli counts were higher in hand-mixed materials (P < 0.05); no other statistically significant differences were found between hand- and auto-mixed materials. According to the Kruskal-Wallis test, significant differences were found between the disinfection procedures (Z > 2.394). The methods used for mixing polyether impression material did not affect bacterial attachment to impression surfaces. In contrast, the disinfection procedure greatly affects decontamination of the impression surface.

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.

Bacterial vaginosis: Etiology and modalities of treatment—A brief note

PubMed Central

Kumar, Nikhil; Behera, Beauty; Sagiri, Sai S.; Pal, Kunal; Ray, Sirsendu S.; Roy, Saroj

2011-01-01

A large women population of the world is suffering from a vaginal infection commonly known as bacterial vaginosis. The disease is associated with the decrease in the lactobacilli count in the vagina. Till date, there is a lack of full proof treatment modalities for the cure of the disease. The treatment includes the use of antimicrobials and/or acidifying agents and probiotics, either separately or in combination. This note discusses about the etiology and the various present-day modalities of treatment of bacterial vaginosis. PMID:22219582

Short-term microbiological effects of scaling and root planing and essential-oils mouthwash in Chinese adults*

PubMed Central

He, Jia-yan; Qi, Gang-gang; Huang, Wu-jing; Sun, Xu-dong; Tong, Yu; Peng, Chun-mei; Zhou, Xue-ping; Chen, Hui

2013-01-01

Objective: To assess the short-term effect of scaling and root planing (SRP) and essential-oils mouthwash on the levels of specific bacteria in Chinese adults. Methods: Fifty Chinese adults with chronic periodontitis were randomly assigned to full-mouth SRP or a 7-d essential-oils mouthwash regimen. In addition, 22 periodontally healthy adults used essential-oils mouthwash for 7 d. Clinical examination and plaque/saliva sampling were performed at baseline and on Day 7. Quantitative real-time polymerase chain reaction (PCR) was used to measure Aggregatibacter actinomycetemcomitans (Aa), Fusobacterium nucleatum (Fn), Porphyromonas gingivalis (Pg), Prevotella intermedia (Pi), and total bacterial loads in saliva, supra- and sub-gingival plaque samples. Results: The detection frequencies of four tested species remained unchanged after either treatment. However, the bacterial loads of Fn, Pg, and Pi were significantly reduced by SRP; the mean reduction of bacterial counts in saliva ranged from 52.2% to 62.5% (p<0.01), in supragingival plaque from 68.2% to 81.0% (p<0.05), and in subgingival plaque from 67.9% to 93.0% (p<0.01). Total bacterial loads were reduced after SRP in supra- and sub-gingival plaque (p<0.05). Essential-oils mouthwash reduced Fn levels in supragingival plaque by a mean of 53.2%, and reduced total bacterial loads in supra- and sub-gingival plaque (p<0.01). In subgingival plaque from periodontal patients, Pg and Pi reductions were high after SRP compared to essential-oils mouthwash (93.0% vs. 37.7% and 87.0% vs. 21.0%, p<0.05). No significant bacterial reduction was observed in periodontally healthy subjects using essential-oils mouthwash. Conclusions: SRP and essential-oils mouthwash both have an impact on saliva and gingival plaque flora in Chinese periodontitis patients in 7 d, with greater microbiological improvement by SRP. PMID:23645178

Multi-parameter flow cytometry as a process analytical technology (PAT) approach for the assessment of bacterial ghost production.

PubMed

Langemann, Timo; Mayr, Ulrike Beate; Meitz, Andrea; Lubitz, Werner; Herwig, Christoph

2016-01-01

Flow cytometry (FCM) is a tool for the analysis of single-cell properties in a cell suspension. In this contribution, we present an improved FCM method for the assessment of E-lysis in Enterobacteriaceae. The result of the E-lysis process is empty bacterial envelopes-called bacterial ghosts (BGs)-that constitute potential products in the pharmaceutical field. BGs have reduced light scattering properties when compared with intact cells. In combination with viability information obtained from staining samples with the membrane potential-sensitive fluorescent dye bis-(1,3-dibutylarbituric acid) trimethine oxonol (DiBAC4(3)), the presented method allows to differentiate between populations of viable cells, dead cells, and BGs. Using a second fluorescent dye RH414 as a membrane marker, non-cellular background was excluded from the data which greatly improved the quality of the results. Using true volumetric absolute counting, the FCM data correlated well with cell count data obtained from colony-forming units (CFU) for viable populations. Applicability of the method to several Enterobacteriaceae (different Escherichia coli strains, Salmonella typhimurium, Shigella flexneri 2a) could be shown. The method was validated as a resilient process analytical technology (PAT) tool for the assessment of E-lysis and for particle counting during 20-l batch processes for the production of Escherichia coli Nissle 1917 BGs.

Stability of saliva microbiota during moderate consumption of red wine.

PubMed

Barroso, Elvira; Martín, Virginia; Martínez-Cuesta, M Carmen; Peláez, Carmen; Requena, Teresa

2015-12-01

This study has evaluated the effect of regular and moderate red wine consumption on the diversity and occurrence of different groups of bacteria that are representative in human saliva. Saliva from twenty-two healthy volunteers (age range 20-48 years) was analyzed in this study. Fourteen individuals consumed red wine (250mL/day) during 4 weeks, whereas 8 volunteers were included in the control group. The evolution and composition of the microbial community in saliva was evaluated by PCR-DGGE and quantitative PCR. The microbial inter-individual variability observed in the PCR-DGGE band patterns was higher than the differences observed after the 4-weeks period of red wine intake. Bifidobacterium dentium, Bifidobacterium spp. and Alloscardovia omnicolens were the most representative bifidobacterial species, whereas the Streptococcus mitis-Streptococcus oralis group predominated within Streptococcus. This genus was the most numerous of the bacterial groups assayed, reaching average counts above 8 log copy numbers/mL. On the other hand, the lowest counts were recorded for Actinomyces, Fusobacterium, Haemophilus, Neisseria and Veillonella, which showed average values of 5 log copy numbers/mL. The results showed no significant differences (P>0.5) in bacterial counts after the period of red wine intake. The overall diversity and stability of representative bacterial groups of the human saliva is not disturbed due to regular-moderate red wine consumption. Copyright © 2015 Elsevier Ltd. All rights reserved.

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).

Quantitative assessment of vaginal microflora during use of tampons of various compositions.

PubMed Central

Onderdonk, A B; Zamarchi, G R; Rodriguez, M L; Hirsch, M L; Muñoz, A; Kass, E H

1987-01-01

Although the effect of vaginal tampons on the microbial flora during menstruation has recently been studied by several investigators, quantitative effects attributable to particular tampon fibers have received less attention. The purposes of the present study were (i) to determine and then to compare the effects of polyacrylate rayon tampons and viscose rayon tampons on the normal vaginal flora, (ii) to compare quantitative bacterial counts obtained from these tampons with those obtained from concomitant vaginal swabs, and (iii) to determine whether either of these tampon types alters the vaginal microflora when compared with the microflora in the same women using all-cotton tampons or external catamenial pads. Tampon and swab samples were obtained at predetermined times from 18 women for an average of seven menstrual cycles. Samples consisting of swabs from women wearing menstrual pads were compared with swab and tampon samples taken at predetermined times during the menstrual cycle from women using cotton, polyacrylate rayon, or viscose rayon tampons. Samples were analyzed for total aerobic, facultative, and anaerobic bacterial counts. Statistical evaluation of the results indicated that, on the whole, total bacterial counts decreased during menstruation and that the numbers of bacteria in tampons tended to be lower than those in swab samples taken at the same time. The tampon type had little effect on the vaginal microflora. PMID:3435142

A NON-DAIRY PROBIOTIC’S (POI) INFLUENCE ON CHANGING THE GASTROINTESTINAL TRACT’S MICROFLORA ENVIRONMENT

PubMed Central

Brown, Amy C.; Shovic, Anne; Ibrahim, Salam; Holck, Peter; Huang, Alvin

2006-01-01

Justification Yogurt has been historically used to restore gut microflora adversely affected by antibiotic treatment. Certain fermented dairy products are probiotics; “live microorganisms which when administered in adequate amounts confer a health benefit to the host.” Microorganisms in foods may benefit certain health conditions such as diarrhea, gastroenteritis, irritable bowel syndrome, inflammatory bowel disease, and cancer. A potential new probiotic from a Polynesian traditional food is poi; a starchy paste made from the corm of taro plants. Objective The purpose of this study was to determine if consumption of poi, a potential non-dairy probiotic, altered the microflora in the gastrointestinal tract of healthy adults. Methods A cross-over clinical study included 18 subjects (19–64 years of age) divided into a poi group (n=10) and control group (n=8). The study duration of 14 weeks consisted of a 2-week washout, 4-week treatment or control, a subsequent 2-week washout, cross-over of 4-week treatment or control, and a final 2-week washout. Subjects thus served as their own controls. While receiving the poi treatment, participants consumed fresh poi (1–2 days old) three times a day (130 g/meal or about _ cup/meal); the control group did not. Both groups filled out 3-day dietary records to ensure compliance. Measurable outcomes include pre-and post-treatment microbiological fecal culture analyses. Results We found no significant differences in total bacterial counts following a poi diet versus following a control diet, nor were significant differences found in counts of specific bacterial species. Lactococcus tends to be higher in poi when it is analyzed for specific bacteria, but the poi consumption in our study did not alter the mean concentration of individual bacterial species (log10 CFU/g wet feces) for Escherichia coli, Enterobacter, Klebsiella, Lactobacillus, Lactococcus, and Bifidobacterium. No significant differences in stool frequency or consistency were observed between the treatment and control group periods. Conclusion Poi consumption did not significantly alter total or individual bacterial counts in the human gastrointestinal tract. Further research might determine if “sour poi” (3–4 days old) has a greater affect than “fresh poi” (1–2 days old) as a potential probiotic, and a larger trial with longer diet durations may detect more subtle effects of poi consumption on bacterial counts. PMID:15712767

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

PubMed

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

2005-01-01

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

Bacterial pneumonia, HIV therapy, and disease progression among HIV-infected women in the HIV epidemiologic research (HER) study.

PubMed

Kohli, Rakhi; Lo, Yungtai; Homel, Peter; Flanigan, Timothy P; Gardner, Lytt I; Howard, Andrea A; Rompalo, Anne M; Moskaleva, Galina; Schuman, Paula; Schoenbaum, Ellie E

2006-07-01

To determine the rate and predictors of community-acquired bacterial pneumonia and its effect on human immunodeficiency virus (HIV) disease progression in HIV-infected women, we performed a multiple-site, prospective study of HIV-infected women in 4 cities in the United States. During the period of 1993-2000, we observed 885 HIV-infected and 425 HIV-uninfected women with a history of injection drug use or high-risk sexual behavior. Participants underwent semiannual interviews, and CD4+ lymphocyte count and viral load were assessed in HIV-infected subjects. Data regarding episodes of bacterial pneumonia were ascertained from medical record reviews. The rate of bacterial pneumonia among 885 HIV-infected women was 8.5 cases per 100 person-years, compared with 0.7 cases per 100 person-years in 425 HIV-uninfected women (P < .001). In analyses limited to follow-up after 1 January 1996, highly active antiretroviral therapy (HAART) and trimethoprim-sulfamethoxazole (TMP-SMX) use were associated with a decreased risk of bacterial pneumonia. Among women who had used TMP-SMX for 12 months, each month of HAART decreased bacterial pneumonia risk by 8% (adjusted hazard ratio [HR(adj)], 0.92; 95% confidence interval [CI], 0.89-0.95). Increments of 50 CD4+ cells/mm3 decreased the risk (HR(adj), 0.88; 95% CI, 0.84-0.93), and smoking doubled the risk (HR(adj), 2.12; 95% CI, 1.26-3.55). Bacterial pneumonia increased mortality risk (HR(adj), 5.02; 95% CI, 2.12-11.87), with adjustment for CD4+ lymphocyte count and duration of HAART and TMP-SMX use. High rates of bacterial pneumonia persist among HIV-infected women. Although HAART and TMP-SMX treatment decreased the risk, bacterial pneumonia was associated with an accelerated progression to death. Interventions that improve HAART utilization and promote smoking cessation among HIV-infected women are warranted.

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.

USE OF SCORE AND CEREBROSPINAL FLUID LACTATE DOSAGE IN DIFFERENTIAL DIAGNOSIS OF BACTERIAL AND ASEPTIC MENINGITIS

PubMed Central

Pires, Frederico Ribeiro; Franco, Andréia Christine Bonotto Farias; Gilio, Alfredo Elias; Troster, Eduardo Juan

2017-01-01

ABSTRACT Objective: To evaluate Bacterial Meningitis Score (BMS) on its own and in association with Cerebrospinal Fluid (CSF) lactate dosage in order to distinguish bacterial from aseptic meningitis. Methods: Children diagnosed with meningitis at a tertiary hospital between January/2011 and December/2014 were selected. All data were obtained upon admission. BMS was applied and included: CSF Gram staining (2 points); CSF neutrophil count ≥1,000 cells/mm3 (1 point); CSF protein ≥80 mg/dL (1 point); peripheral blood neutrophil count ≥10,000 cells/mm3 (1 point) and seizures upon/before arrival (1 point). Cutoff value for CSF lactate was ≥30 mg/dL. Sensitivity, specificity and negative predictive value of several BMS cutoffs and BMS associated with high CSF lactate were evaluated for prediction of bacterial meningitis. Results: Among 439 eligible patients, 94 did not have all data available to complete the score, and 345 patients were included: 7 in bacterial meningitis group and 338 in aseptic meningitis group. As predictive factors of bacterial meningitis, BMS ≥1 had 100% sensitivity (95%CI 47.3-100), 64.2% specificity (58.8-100) and 100% negative predictive value (97.5-100); BMS ≥2 or BMS ≥1 associated with high CSF lactate also showed 100% sensitivity (47.3-100); but 98.5% specificity (96.6-99.5) and 100% negative predictive value (98.3-100). Conclusions: 2 point BMS in association with CSF lactate dosage had the same sensitivity and negative predictive value, with increased specificity for diagnosis of bacterial meningitis when compared with 1-point BMS. PMID:29185620

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

Potentially pathogenic airway bacteria and neutrophilic inflammation in treatment resistant severe asthma.

PubMed

Green, Benjamin J; Wiriyachaiporn, Surasa; Grainge, Christopher; Rogers, Geraint B; Kehagia, Valia; Lau, Laurie; Carroll, Mary P; Bruce, Kenneth D; Howarth, Peter H

2014-01-01

Molecular microbiological analysis of airway samples in asthma has demonstrated an altered microbiome in comparison to healthy controls. Such changes may have relevance to treatment-resistant severe asthma, particularly those with neutrophilic airway inflammation, as bacteria might be anticipated to activate the innate immune response, a process that is poorly steroid responsive. An understanding of the relationship between airway bacterial presence and dominance in severe asthma may help direct alternative treatment approaches. We aimed to use a culture independent analysis strategy to describe the presence, dominance and abundance of bacterial taxa in induced sputum from treatment resistant severe asthmatics and correlate findings with clinical characteristics and airway inflammatory markers. Induced sputum was obtained from 28 stable treatment-resistant severe asthmatics. The samples were divided for supernatant IL-8 measurement, cytospin preparation for differential cell count and Terminal Restriction Fragment Length Polymorphism (T-RFLP) profiling for bacterial community analysis. In 17/28 patients, the dominant species within the airway bacterial community was Moraxella catarrhalis or a member of the Haemophilus or Streptococcus genera. Colonisation with these species was associated with longer asthma disease duration (mean (SD) 31.8 years (16.7) vs 15.6 years (8.0), p = 0.008), worse post-bronchodilator percent predicted FEV1 (68.0% (24.0) vs 85.5% (19.7), p = 0.025) and higher sputum neutrophil differential cell counts (median (IQR) 80% (67-83) vs 43% (29-67), p = 0.001). Total abundance of these organisms significantly and positively correlated with sputum IL-8 concentration and neutrophil count. Airway colonisation with potentially pathogenic micro-organisms in asthma is associated with more severe airways obstruction and neutrophilic airway inflammation. This altered colonisation may have a role in the development of an asthma phenotype that responds less well to current asthma therapies.

Abiotic Stress and Phyllosphere Bacteria Influence the Survival of Human Norovirus and Its Surrogates on Preharvest Leafy Greens

PubMed Central

Esseili, Malak A.; Gao, Xiang; Tegtmeier, Sarah; Saif, Linda J.

2015-01-01

Foodborne outbreaks of human noroviruses (HuNoVs) are frequently associated with leafy greens. Because there is no effective method to eliminate HuNoV from postharvest leafy greens, understanding virus survival under preharvest conditions is crucial. The objective of this study was to evaluate the survival of HuNoV and its surrogate viruses, murine norovirus (MNV), porcine sapovirus (SaV), and Tulane virus (TV), on preharvest lettuce and spinach that were subjected to abiotic stress (physical damage, heat, or flood). We also examined the bacteria culturable from the phyllosphere in response to abiotic stress and in relation to viral persistence. Mature plants were subjected to stressors 2 days prior to inoculation of the viruses on leaves. We quantified the viral RNA, determined the infectivity of the surrogates, and performed bacterial counts on postinoculation days (PIDs) 0, 1, 7, and 14. For both plant types, time exerted significant effects on HuNoV, MNV, SaV, and TV RNA titers, with greater effects being seen for the surrogates. Infectious surrogate viruses were undetectable on PID 14. Only physical damage on PID 14 significantly enhanced HuNoV RNA persistence on lettuce, while the three stressors differentially enhanced the persistence of MNV and TV RNA. Bacterial counts were significantly affected by time and plant type but not by the stressors. However, bacterial counts correlated significantly with HuNoV RNA titers on spinach and with the presence of surrogate viruses on both plant types under various conditions. In conclusion, abiotic stressors and phyllosphere bacterial density may differentially influence the survival of HuNoV and its surrogates on lettuce and spinach, emphasizing the need for the use of preventive measures at the preharvest stage. PMID:26497461

The short-term treatment effects on the microbiota at the dorsum of the tongue in intra-oral halitosis patients--a randomized clinical trial.

PubMed

Ademovski, Seida Erovic; Persson, G Rutger; Winkel, Edwin; Tangerman, Albert; Lingström, Peter; Renvert, Stefan

2013-03-01

This study aims to assess the effects of rinsing with zinc- and chlorhexidine-containing mouth rinse with or without adjunct tongue scraping on volatile sulfur compounds (VSCs) in breath air, and the microbiota at the dorsum of the tongue. A randomized single-masked controlled clinical trial with a cross-over study design over 14 days including 21 subjects was performed. Bacterial samples from the dorsum of the tongue were assayed by checkerboard DNA-DNA hybridization. No halitosis (identified by VSC assessments) at day 14 was identified in 12/21 subjects with active rinse alone, in 10/21 with adjunct use of tongue scraper, in 1/21 for negative control rinse alone, and in 3/21 in the control and tongue scraping sequence. At day 14, significantly lower counts were identified only in the active rinse sequence (p < 0.001) for 15/78 species including, Fusobacterium sp., Porphyromonas gingivalis, Pseudomonas aeruginosa, Staphylococcus aureus, and Tannerella forsythia. A decrease in bacteria from baseline to day 14 was found in successfully treated subjects for 9/74 species including: P. gingivalis, Prevotella melaninogenica, S. aureus, and Treponema denticola. Baseline VSC scores were correlated with several bacterial species. The use of a tongue scraper combined with active rinse did not change the levels of VSC compared to rinsing alone. VSC scores were not associated with bacterial counts in samples taken from the dorsum of the tongue. The active rinse alone containing zinc and chlorhexidine had effects on intra-oral halitosis and reduced bacterial counts of species associated with malodor. Tongue scraping provided no beneficial effects on the microbiota studied. Periodontally healthy subjects with intra-oral halitosis benefit from daily rinsing with zinc- and chlorhexidine-containing mouth rinse.

Community-acquired bacterial pneumonia in human immunodeficiency virus-infected patients: validation of severity criteria. The Grupo Andaluz para el Estudio de las Enfermedades Infecciosas.

PubMed

Cordero, E; Pachón, J; Rivero, A; Girón, J A; Gómez-Mateos, J; Merino, M D; Torres-Tortosa, M; González-Serrano, M; Aliaga, L; Collado, A; Hernández-Quero, J; Barrera, A; Nuño, E

2000-12-01

Severity criteria for community-acquired pneumonia (CAP) have always excluded patients with human immunodeficiency virus (HIV) infection. A 1-yr, multicenter, prospective observational study of HIV-infected patients with bacterial CAP was done to validate the criteria used in the American Thoracic Society (ATS) guidelines for CAP, and to determine the prognosis-associated factors in the HIV-infected population with bacterial CAP. Overall, 355 cases were included, with an attributable mortality of 9.3%. Patients who met the ATS criteria had a longer hospital stay (p = 0.01), longer duration of fever (p < 0.001), and higher attributable mortality (13.1% versus 3.5%, p = 0.02) than those who did not. Three factors were independently related to mortality: CD4(+) cell count < 100/microl, radiologic progression of disease, and shock. Pleural effusion, cavities, and/or multilobar infiltrates at admission were independently associated with radiologic progression. A prognostic rule based on the five criteria of shock, CD4(+) cell count < 100/microl, pleural effusion, cavities, and multilobar infiltrates had a high negative predictive value for mortality (97.1%). The attributable mortality for severe pneumonia was 11.3%, as compared with 1.3% for nonsevere disease (p = 0.008). The ATS severity criteria are valid in HIV-infected patients with bacterial CAP. Our study provides the basis for identification of patients who may require hospitalization determined by clinical judgment and the five clinical criteria of shock, a CD4(+) cell count < 100/microl, pleural effusion, cavities, and multilobar involvement. These prognostic factors should be validated in independent cohort studies.

Effect of compliance during periodontal maintenance therapy on levels of bacteria associated with periodontitis: A 6-year prospective study.

PubMed

Costa, Fernando Oliveira; Vieira, Thaís Riberal; Cortelli, Sheila Cavalca; Cota, Luís Otávio Miranda; Costa, José Eustáquio; Aguiar, Maria Cássia Ferreira; Cortelli, José Roberto

2018-05-01

It is well established that regular compliance during periodontal maintenance therapy (PMT) maintains the stability of periodontal clinical parameters obtained after active periodontal therapy (APT). However, compliance during PMT has not yet been related to subgingival bacterial levels. Thus, this study followed individuals in PMT over 6 years and longitudinally evaluated the effects of compliance on periodontitis-associated bacterial levels and its relation to periodontal status. From a 6-year prospective cohort study with 212 individuals in PMT, 91 were determined to be eligible. From this total, 28 regular compliers (RC) were randomly selected and matched for age and sex with 28 irregular compliers (IC). Complete periodontal examination and microbiological samples were obtained 5 times: T1 (prior to APT), T2 (after APT), T3 (2 years), T4 (4 years), and T5 (6 years). Total bacteria counts and levels of Actinomyces naeslundii, Porphyromonas gingivalis, Tannerella forsythia, and Treponema denticola were evaluated through quantitative polymerase chain reaction. RC had less tooth loss and better clinical and microbiological conditions over time when compared with IC. IC had higher total bacterial counts and higher levels of T. denticola. Moreover, among IC, total bacterial counts were positively associated with plaque index and bleeding on probing, while levels of A. naeslundii, T. forsythia, and T. denticola were negatively associated with clinical attachment loss (4 to 5 mm) among RC. Compliance positively influenced subgingival microbiota and contributed to stability of periodontal clinical status. Regular visits during PMT sustained microbiological benefits provided by APT over a 6-year period. © 2018 American Academy of Periodontology.

Prophylactic Effect of a Therapeutic Vaccine against TB Based on Fragments of Mycobacterium tuberculosis

PubMed Central

Cáceres, Neus; Pinto, Sergio; Díaz, Jorge; Cardona, Pere-Joan

2011-01-01

The prophylactic capacity of the RUTI® vaccine, based on fragmented cells of Mycobacterium tuberculosis, has been evaluated in respect to aerosol challenge with virulent bacilli. Subcutaneous vaccination significantly reduced viable bacterial counts in both lungs and spleens of C57Bl mice, when challenged 4 weeks after vaccination. RUTI® protected the spleen less than BCG. Following a 9 month vaccination-challenge interval, protection was observed for the lungs, but not for the spleen. Survival of infected guinea pigs was prolonged by vaccination given 5 weeks before challenge. Inoculations of RUTI® shortly after infection significantly reduced the viable bacterial counts in the lungs, when compared with infected control mice. Thus, vaccination by RUTI® has potential for both the prophylaxis and immunotherapy of tuberculosis. PMID:21647222

Prophylactic effect of a therapeutic vaccine against TB based on fragments of Mycobacterium tuberculosis.

PubMed

Vilaplana, Cristina; Gil, Olga; Cáceres, Neus; Pinto, Sergio; Díaz, Jorge; Cardona, Pere-Joan

2011-01-01

The prophylactic capacity of the RUTI® vaccine, based on fragmented cells of Mycobacterium tuberculosis, has been evaluated in respect to aerosol challenge with virulent bacilli. Subcutaneous vaccination significantly reduced viable bacterial counts in both lungs and spleens of C57Bl mice, when challenged 4 weeks after vaccination. RUTI® protected the spleen less than BCG. Following a 9 month vaccination-challenge interval, protection was observed for the lungs, but not for the spleen. Survival of infected guinea pigs was prolonged by vaccination given 5 weeks before challenge. Inoculations of RUTI® shortly after infection significantly reduced the viable bacterial counts in the lungs, when compared with infected control mice. Thus, vaccination by RUTI® has potential for both the prophylaxis and immunotherapy of tuberculosis.

Mortality in Severe Human Immunodeficiency Virus-Tuberculosis Associates With Innate Immune Activation and Dysfunction of Monocytes

PubMed Central

Janssen, Saskia; Schutz, Charlotte; Ward, Amy; Nemes, Elisa; Wilkinson, Katalin A; Scriven, James; Huson, Mischa A; Aben, Nanne; Maartens, Gary; Burton, Rosie; Wilkinson, Robert J; Grobusch, Martin P; Van der Poll, Tom; Meintjes, Graeme

2017-01-01

Abstract Background Case fatality rates among hospitalized patients diagnosed with human immunodeficiency virus (HIV)-associated tuberculosis remain high, and tuberculosis mycobacteremia is common. Our aim was to define the nature of innate immune responses associated with 12-week mortality in this population. Methods This prospective cohort study was conducted at Khayelitsha Hospital, Cape Town, South Africa. Hospitalized HIV-infected tuberculosis patients with CD4 counts <350 cells/µL were included; tuberculosis blood cultures were performed in all. Ambulatory HIV-infected patients without active tuberculosis were recruited as controls. Whole blood was stimulated with Escherichia coli derived lipopolysaccharide, heat-killed Streptococcus pneumoniae, and Mycobacterium tuberculosis. Biomarkers of inflammation and sepsis, intracellular (flow cytometry) and secreted cytokines (Luminex), were assessed for associations with 12-week mortality using Cox proportional hazard models. Second, we investigated associations of these immune markers with tuberculosis mycobacteremia. Results Sixty patients were included (median CD4 count 53 cells/µL (interquartile range [IQR], 22–132); 16 (27%) died after a median of 12 (IQR, 0–24) days. Thirty-one (52%) grew M. tuberculosis on blood culture. Mortality was associated with higher concentrations of procalcitonin, activation of the innate immune system (% CD16+CD14+ monocytes, interleukin-6, tumour necrosis factor-ɑ and colony-stimulating factor 3), and antiinflammatory markers (increased interleukin-1 receptor antagonist and lower monocyte and neutrophil responses to bacterial stimuli). Tuberculosis mycobacteremia was not associated with mortality, nor with biomarkers of sepsis. Conclusions Twelve-week mortality was associated with greater pro- and antiinflammatory alterations of the innate immune system, similar to those reported in severe bacterial sepsis. PMID:28369200

Mortality in Severe Human Immunodeficiency Virus-Tuberculosis Associates With Innate Immune Activation and Dysfunction of Monocytes.

PubMed

Janssen, Saskia; Schutz, Charlotte; Ward, Amy; Nemes, Elisa; Wilkinson, Katalin A; Scriven, James; Huson, Mischa A; Aben, Nanne; Maartens, Gary; Burton, Rosie; Wilkinson, Robert J; Grobusch, Martin P; Van der Poll, Tom; Meintjes, Graeme

2017-07-01

Case fatality rates among hospitalized patients diagnosed with human immunodeficiency virus (HIV)-associated tuberculosis remain high, and tuberculosis mycobacteremia is common. Our aim was to define the nature of innate immune responses associated with 12-week mortality in this population. This prospective cohort study was conducted at Khayelitsha Hospital, Cape Town, South Africa. Hospitalized HIV-infected tuberculosis patients with CD4 counts <350 cells/µL were included; tuberculosis blood cultures were performed in all. Ambulatory HIV-infected patients without active tuberculosis were recruited as controls. Whole blood was stimulated with Escherichia coli derived lipopolysaccharide, heat-killed Streptococcus pneumoniae, and Mycobacterium tuberculosis. Biomarkers of inflammation and sepsis, intracellular (flow cytometry) and secreted cytokines (Luminex), were assessed for associations with 12-week mortality using Cox proportional hazard models. Second, we investigated associations of these immune markers with tuberculosis mycobacteremia. Sixty patients were included (median CD4 count 53 cells/µL (interquartile range [IQR], 22-132); 16 (27%) died after a median of 12 (IQR, 0-24) days. Thirty-one (52%) grew M. tuberculosis on blood culture. Mortality was associated with higher concentrations of procalcitonin, activation of the innate immune system (% CD16+CD14+ monocytes, interleukin-6, tumour necrosis factor-ɑ and colony-stimulating factor 3), and antiinflammatory markers (increased interleukin-1 receptor antagonist and lower monocyte and neutrophil responses to bacterial stimuli). Tuberculosis mycobacteremia was not associated with mortality, nor with biomarkers of sepsis. Twelve-week mortality was associated with greater pro- and antiinflammatory alterations of the innate immune system, similar to those reported in severe bacterial sepsis. © The Author 2017. Published by Oxford University Press for the Infectious Diseases Society of America.

Automated Image Analysis for the Detection of Benthic Crustaceans and Bacterial Mat Coverage Using the VENUS Undersea Cabled Network

PubMed Central

Aguzzi, Jacopo; Costa, Corrado; Robert, Katleen; Matabos, Marjolaine; Antonucci, Francesca; Juniper, S. Kim; Menesatti, Paolo

2011-01-01

The development and deployment of sensors for undersea cabled observatories is presently biased toward the measurement of habitat variables, while sensor technologies for biological community characterization through species identification and individual counting are less common. The VENUS cabled multisensory network (Vancouver Island, Canada) deploys seafloor camera systems at several sites. Our objective in this study was to implement new automated image analysis protocols for the recognition and counting of benthic decapods (i.e., the galatheid squat lobster, Munida quadrispina), as well as for the evaluation of changes in bacterial mat coverage (i.e., Beggiatoa spp.), using a camera deployed in Saanich Inlet (103 m depth). For the counting of Munida we remotely acquired 100 digital photos at hourly intervals from 2 to 6 December 2009. In the case of bacterial mat coverage estimation, images were taken from 2 to 8 December 2009 at the same time frequency. The automated image analysis protocols for both study cases were created in MatLab 7.1. Automation for Munida counting incorporated the combination of both filtering and background correction (Median- and Top-Hat Filters) with Euclidean Distances (ED) on Red-Green-Blue (RGB) channels. The Scale-Invariant Feature Transform (SIFT) features and Fourier Descriptors (FD) of tracked objects were then extracted. Animal classifications were carried out with the tools of morphometric multivariate statistic (i.e., Partial Least Square Discriminant Analysis; PLSDA) on Mean RGB (RGBv) value for each object and Fourier Descriptors (RGBv+FD) matrices plus SIFT and ED. The SIFT approach returned the better results. Higher percentages of images were correctly classified and lower misclassification errors (an animal is present but not detected) occurred. In contrast, RGBv+FD and ED resulted in a high incidence of records being generated for non-present animals. Bacterial mat coverage was estimated in terms of Percent Coverage and Fractal Dimension. A constant Region of Interest (ROI) was defined and background extraction by a Gaussian Blurring Filter was performed. Image subtraction within ROI was followed by the sum of the RGB channels matrices. Percent Coverage was calculated on the resulting image. Fractal Dimension was estimated using the box-counting method. The images were then resized to a dimension in pixels equal to a power of 2, allowing subdivision into sub-multiple quadrants. In comparisons of manual and automated Percent Coverage and Fractal Dimension estimates, the former showed an overestimation tendency for both parameters. The primary limitations on the automatic analysis of benthic images were habitat variations in sediment texture and water column turbidity. The application of filters for background corrections is a required preliminary step for the efficient recognition of animals and bacterial mat patches. PMID:22346657

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.

Compositions of maple sap microflora and collection system biofilms evaluated by scanning electron microscopy and denaturing gradient gel electrophoresis.

PubMed

Lagacé, L; Jacques, M; Mafu, A A; Roy, D

2006-05-25

The bacterial microflora of maple sap and biofilms in collection system tubing were studied through the use of bacterial counts, scanning electron microscopy (SEM) of surfaces and the analysis of 16S rRNA gene by denaturing gradient gel electrophoresis (DGGE). Samples were taken at five times during the 2002 and 2003 seasons in order to follow the changes in the microflora of this complex ecosystem. Bacterial counts showed the growth of bacterial populations as the season advanced. These populations were mainly composed of psychrotrophic bacteria and Pseudomonas spp. SEM results confirmed the suspected presence of biofilms on the inner surfaces of tubing samples. Bacterial colonization and biofilm formation progressively increased during the season for both lateral and main line surfaces, and biofilms were mainly composed of rod shape bacteria. The bacterial microflora profiles obtained for sap and corresponding biofilm by DGGE showed up to 12 major bands. The Shannon-Weaver index of diversity (H) calculated from DGGE bands were statistically higher for sap samples compared to biofilm. The diversity index was relatively stable or increasing for lateral line sap and biofilm samples during the season while the diversity index for sap and biofilm samples of the main line showed a decreasing profile as the season progressed. Sequence analysis of major DGGE bands revealed the predominance of bacteria from the genera Pseudomonas, Rahnella and another, unidentified genus. The results describe the composition of sap collection system microflora as well as the formation of biofilms and will be useful for further studies on factors affecting maple product quality.

Role of type 1 and S fimbriae in the pathogenesis of Escherichia coli O18:K1 bacteremia and meningitis in the infant rat.

PubMed

Saukkonen, K M; Nowicki, B; Leinonen, M

1988-04-01

The role of fimbriae in the pathogenesis of Escherichia coli infection was studied in the infant rat model. Rat pups were challenged intraperitoneally at the age of 5 days with E. coli K1 (strain IH3080, O18:K1:H7) and three different subpopulations (type 1, type S, or nonfimbriated) of it. All bacterial subpopulations were able to produce peritonitis, bacteremia, and meningitis. However, the type 1 fraction was the least virulent and the type S fraction was the most virulent, as judged by the bacterial counts in body fluids and by the mortality rates of the pups. Fimbrial phase variation to mainly the type-S-fimbriated forms was observed in all body fluids. An initially type-S-fimbriated inoculum remained predominantly type S fimbriated in the peritoneal fluid and blood. In the cerebrospinal fluid, however, about 50% of the bacteria were type S fimbriated and 50% were nonfimbriated 1 h after challenge with the type-S-fimbriated subpopulation; at later times the share of type-S-fimbriated bacteria also increased in the cerebrospinal fluid.

Characterization of bacterial pathogens in rural and urban irrigation water.

PubMed

Aijuka, Matthew; Charimba, George; Hugo, Celia J; Buys, Elna M

2015-03-01

The study aimed to compare the bacteriological quality of an urban and rural irrigation water source. Bacterial counts, characterization, identification and diversity of aerobic bacteria were determined. Escherichia coli isolated from both sites was subjected to antibiotic susceptibility testing, virulence gene (Stx1/Stx2 and eae) determination and (GTG)5 Rep-PCR fingerprinting. Low mean monthly counts for aerobic spore formers, anaerobic spore formers and Staphylococcus aureus were noted although occasional spikes were observed. The most prevalent bacterial species at both sites were Bacillus spp., E. coli and Enterobacter spp. In addition, E. coli and Bacillus spp. were most prevalent in winter and summer respectively. Resistance to at least one antibiotic was 84% (rural) and 83% (urban). Highest resistance at both sites was to cephalothin and ampicillin. Prevalence of E. coli possessing at least one virulence gene (Stx1/Stx2 and eae) was 15% (rural) and 42% (urban). All (rural) and 80% (urban) of E. coli possessing virulence genes showed antibiotic resistance. Complete genetic relatedness (100%) was shown by 47% of rural and 67% of urban E. coli isolates. Results from this study show that surface irrigation water sources regardless of geographical location and surrounding land-use practices can be reservoirs of similar bacterial pathogens.

Characterization of Early Microbial Communities on Volcanic Deposits along a Vegetation Gradient on the Island of Miyake, Japan

PubMed Central

Guo, Yong; Fujimura, Reiko; Sato, Yoshinori; Suda, Wataru; Kim, Seok-won; Oshima, Kenshiro; Hattori, Masahira; Kamijo, Takashi; Narisawa, Kazuhiko; Ohta, Hiroyuki

2014-01-01

The 2000 eruption of Mount Oyama on the island of Miyake (Miyake-jima) created a unique opportunity to study the early ecosystem development on newly exposed terrestrial substrates. In this study, bacterial and fungal communities on 9- and 11-year-old volcanic deposits at poorly to fully vegetation-recovered sites in Miyake-jima, Japan, were characterized by conventional culture-based methods and pyrosequencing of 16S rRNA and 18S rRNA genes. Despite the differences in the vegetation cover, the upper volcanic deposit layer samples displayed low among-site variation for chemical properties (pH, total organic carbon, and total nitrogen) and microbial population densities (total direct count and culturable count). Statistical analyses of pyrosequencing data revealed that the microbial communities of volcanic deposit samples were phylogenetically diverse, in spite of very low-carbon environmental conditions, and their diversity was comparable to that in the lower soil layer (buried soil) samples. Comparing with the microbial communities in buried soil, the volcanic deposit communities were characterized by the presence of Betaproteobacteria and Gammaproteobacteria as the main bacterial class, Deinococcus- Thermus as the minor bacterial phyla, and Ascomycota as the major fungal phyla. Multivariate analysis revealed that several bacterial families and fungal classes correlated positively or negatively with plant species. PMID:24463576

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

Evaluation of antimicrobial efficacy of Aloe vera and Meswak containing dentifrices with fluoridated dentifrice: An in vivo study

PubMed Central

Bhati, Neha; Jaidka, Shipra; Somani, Rani

2015-01-01

Aim: To comparatively evaluate the antimicrobial efficacy of fluoridated and herbal dentifrices. Materials and Methods: Sixty students in the age group 6–12 years with DMF/def score 0 were selected from an orphanage center. The participants were divided into four groups. In group A, no dentifrice was used; in group B, fluoride containing dentifrice was used; group C subjects used Aloe vera containing dentifrice; and in group D, Meswak containing dentifrice was used. The salivary samples were collected at the washout period of 2 days, 15 days, and 30 days and cultured on Mitis Salivarius Agar for determining Streptococcus mutans count. Results obtained were statistically analyzed using Student's t-test. Results: There was an increase in bacterial count in group A where no dentifrices were used, while the bacterial count steadily decreased in groups B, C, and D by 83.7%, 80.94%, and 83.5%, respectively. Conclusion: Herbal dentifrices containing A. vera and Meswak can be safely recommended as an alternative to fluoridated dentifrices in terms of antimicrobial efficacy. PMID:26539392

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.

Microbial profile on metallic and ceramic bracket materials.

PubMed

Anhoury, Patrick; Nathanson, Dan; Hughes, Christopher V; Socransky, Sigmund; Feres, Magda; Chou, Laisheng Lee

2002-08-01

The placement of orthodontic appliances creates a favorable environment for the accumulation of a microbiota and food residues, which, in time, may cause caries or exacerbate any pre-existing periodontal disease. The purpose of the present study was to compare the total bacterial counts present on metallic and ceramic orthodontic brackets in order to clarify which bracket type has a higher plaque retaining capacity and to determine the levels of Streptococcus mutans and Lactobacillus spp on both types of brackets. Thirty-two metallic brackets and 24 ceramic brackets were collected from orthodontic patients at the day of debonding. Two brackets were collected from each patient; one from a maxillary central incisor and another from a maxillary second premolar. Sixteen patients who used metallic brackets and 12 patients who used ceramic brackets were sampled. Bacterial populations were studied using "checkerboard" DNA-DNA hybridization, which uses DNA probes to identify species in complex microbial samples. The significance of differences between groups was determined using the Mann-Whitney U-test. Results showed no significant differences between metallic and ceramic brackets with respect to the caries-inducing S mutans and L acidophilus spp counts. Mean counts of 8 of 35 additional species differed significantly between metallic and ceramic brackets with no obvious pattern favoring one bracket type over the other. This study showed higher mean counts of Treponema denticola, Actinobacillus actinomycetemcomitans, Fusobacterium nucleatum ss vincentii, Streptococcus anginosus, and Eubacterium nodatum on metallic brackets while higher counts of Eikenella corrodens, Campylobacter showae, and Selenomonas noxia were found on ceramic brackets.

Zoonotic bacterial meningitis in human adults.

PubMed

van Samkar, Anusha; Brouwer, Matthijs C; van der Ende, Arie; van de Beek, Diederik

2016-09-13

To describe the epidemiology, etiology, clinical characteristics, treatment, outcome, and prevention of zoonotic bacterial meningitis in human adults. We identified 16 zoonotic bacteria causing meningitis in adults. Zoonotic bacterial meningitis is uncommon compared to bacterial meningitis caused by human pathogens, and the incidence has a strong regional distribution. Zoonotic bacterial meningitis is mainly associated with animal contact, consumption of animal products, and an immunocompromised state of the patient. In a high proportion of zoonotic bacterial meningitis cases, CSF analysis showed only a mildly elevated leukocyte count. The recommended antibiotic therapy differs per pathogen, and the overall mortality is low. Zoonotic bacterial meningitis is uncommon but is associated with specific complications. The suspicion should be raised in patients with bacterial meningitis who have recreational or professional contact with animals and in patients living in regions endemic for specific zoonotic pathogens. An immunocompromised state is associated with a worse prognosis. Identification of risk factors and underlying disease is necessary to improve treatment. © 2016 American Academy of Neurology.

Asthma Quiz

MedlinePlus

... the AAAAI Foundation Donate Utility navigation Español Journals Pollen Counts Annual Meeting Member Login / My Membership Search ... bacterial infections, exercise, exposure to allergens such as pollen, dust mites or pet dander, acid reflux, some ...

Preoperative Hand Decontamination in Ophthalmic Surgery: A Comparison of the Removal of Bacteria from Surgeons' Hands by Routine Antimicrobial Scrub versus an Alcoholic Hand Rub.

PubMed

Forer, Yaara; Block, Colin; Frenkel, Shahar

2017-09-01

The goal of this experiment was to evaluate and compare the antimicrobial efficacy of routine preoperative hand washing using commercial medicated sponge brushes versus an alcoholic hand rub, by comparing bacterial growth on ophthalmic surgeons' hands after application of each of these methods. Twenty ophthalmic surgeons were recruited at the Hadassah-Hebrew University Medical Center in Jerusalem, Israel. Samples were collected twice from the hands of each surgeon after hand decontamination using two different protocols during routine surgical practice. The routine preparation consisted of a 3-minute surgical scrub using commercial brush-sponges incorporating either 4% chlorhexidine gluconate (CHG) or 1% povidone-iodine (PVP-I) formulations with detergent, followed by drying the hands with a sterile towel, while the 70% ethanol solution was applied for 60-seconds and allowed to air dry. Half of the group was randomly assigned to provide samples first after the routine method and the alcoholic solution a week later, and the other half of the group was sampled in the reverse order. Viable counts of bacteria were evaluated using a modified glove juice method. Bacterial colonies were enumerated after incubation for 24 hours and expressed as colony forming units (CFU)/mL for each pair of hands. Geometric mean counts were 1310 and 39 CFU/mL, in the routine and alcohol rub groups, respectively, representing a mean log 10 reduction in 1.53. The difference between the paired bacterial counts for the routine versus the alcohol rub was statistically significant (p < 0.0001). There was no statistically significant difference between log 10 reductions for CHG and PVP-I (p = 0.97). This study provides evidence that an alcohol rub protocol is more effective in reducing bacterial counts on hands than routine surgical hand preparation with PVP-I and CHG in a population of practicing ophthalmic surgeons in the operative clinical setting. Thus, it provides a safe alternative as a preoperative hand disinfection method.

Randomized Controlled Trial of Antiseptic Hand Hygiene Methods in an Outpatient Surgery Clinic.

PubMed

Therattil, Paul J; Yueh, Janet H; Kordahi, Anthony M; Cherla, Deepa V; Lee, Edward S; Granick, Mark S

2015-12-01

Outpatient wound care plays an integral part in any plastic surgery practice. However, compliance with hand hygiene measures has shown to be low, due to skin irritation and lack of time. The objective of this trial was to determine whether single-use, long-acting antiseptics can be as effective as standard multiple-use hand hygiene methods in an outpatient surgical setting. A prospective, randomized controlled trial was performed in the authors' outpatient plastic surgery clinic at Rutgers New Jersey Medical School, Newark, NJ to compare the efficacy of an ethyl alcohol-based sanitizer (Avagard D Instant Hand Aniseptic, 3M Health Care, St. Paul, MN), a benzalkonium chloride-based sanitizer (Soft & Shield, Bioderm Technologies, Inc, Trenton, NJ, distributed by NAPP Technologies, Hackensack, NJ ), and soap and- water handwashing. Subjects included clinic personnel, who were followed throughout the course of a 3-hour clinic session with hourly hand bacterial counts taken. During the course of the trial, 95 subjects completed the clinic session utilizing 1 of the hand hygiene methods (36 ethyl alcohol-based sanitizer, 38 benzalkonium chloride-based sanitizer, and 21 soap-and-water handwashing). There was no difference between hand bacterial counts using the different methods at 4 hourly time points (P greater than 0.05). Hand bacterial counts increased significantly over the 3-hour clinic session with the ethyl alcohol-based sanitizer (9.24 to 21.90 CFU, P less than 0.05), benzalkonium chloride-based sanitizer (6.69 to 21.59 CFU, P less than 0.05), and soap-and-water handwashing (8.43 to 22.75 CFU, P less than 0.05). There does not appear to be any difference in efficacy between single-use, long-acting sanitizer, and standard multiple-use hand hygiene methods. Hand bacterial counts increased significantly over the course of the 3-hour clinic session regardless of the hand hygiene measure used. Hand condition of subjects was improved with the ethyl alcohol-based sanitizer and the benzalkonium chloride-based sanitizer compared with soap-and-water handwashing.

Bacterial prostatitis.

PubMed

Gill, Bradley C; Shoskes, Daniel A

2016-02-01

The review provides the infectious disease community with a urologic perspective on bacterial prostatitis. Specifically, the article briefly reviews the categorization of prostatitis by type and provides a distillation of new findings published on bacterial prostatitis over the past year. It also highlights key points from the established literature. Cross-sectional prostate imaging is becoming more common and may lead to more incidental diagnoses of acute bacterial prostatitis. As drug resistance remains problematic in this condition, the reemergence of older antibiotics such as fosfomycin, has proven beneficial. With regard to chronic bacterial prostatitis, no clear clinical risk factors emerged in a large epidemiological study. However, bacterial biofilm formation has been associated with more severe cases. Surgery has a limited role in bacterial prostatitis and should be reserved for draining of a prostatic abscess or the removal of infected prostatic stones. Prostatitis remains a common and bothersome clinical condition. Antibiotic therapy remains the basis of treatment for both acute and chronic bacterial prostatitis. Further research into improving prostatitis treatment is indicated.

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.

Comparison of innovative molecular approaches and standard spore assays for assessment of surface cleanliness.

PubMed

Cooper, Moogega; La Duc, Myron T; Probst, Alexander; Vaishampayan, Parag; Stam, Christina; Benardini, James N; Piceno, Yvette M; Andersen, Gary L; Venkateswaran, Kasthuri

2011-08-01

A bacterial spore assay and a molecular DNA microarray method were compared for their ability to assess relative cleanliness in the context of bacterial abundance and diversity on spacecraft surfaces. Colony counts derived from the NASA standard spore assay were extremely low for spacecraft surfaces. However, the PhyloChip generation 3 (G3) DNA microarray resolved the genetic signatures of a highly diverse suite of microorganisms in the very same sample set. Samples completely devoid of cultivable spores were shown to harbor the DNA of more than 100 distinct microbial phylotypes. Furthermore, samples with higher numbers of cultivable spores did not necessarily give rise to a greater microbial diversity upon analysis with the DNA microarray. The findings of this study clearly demonstrated that there is not a statistically significant correlation between the cultivable spore counts obtained from a sample and the degree of bacterial diversity present. Based on these results, it can be stated that validated state-of-the-art molecular techniques, such as DNA microarrays, can be utilized in parallel with classical culture-based methods to further describe the cleanliness of spacecraft surfaces.

Growth and heavy metal removal by Klebsiella aerogenes at different pH and temperature

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

Al-Shahwani, M.F.; Jazrawi, S.F.; Al-Rawi, E.H.

1984-01-01

A strain of Klebsiella aerogenes isolated from Rustamiyah Station for treatment of wastewater was examined for its ability to grow in a media supplemented with maximum tolerance concentrations of Pb/sup + +/, Zn/sup + +/, Ni/sup + +/, and Cd/sup + +/, separately, at different temperatures and initial pH. The results indicated that at 28/sup 0/C during the first 24 hr, Pb/sup + +/ and Ni/sup + +/ had no effect on the growth of the bacteria, while the presence of Zn/sup + +/ and Cd/sup + +/ decreased the cell count. The growth reached a maximum level after themore » second day and started to decrease gradually. The bacterial count at 37/sup 0/C was less than that at 28/sup 0/C. No bacterial multiplication occurred at 44/sup 0/C. There was little difference between heavy metal removal at 28 and 37/sup 0/C. At 44/sup 0/C, little removal took place. In general, slightly acidic or neutral medium was better for both bacterial growth and metal removal.« less

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.

Impact of pulsed xenon ultraviolet disinfection on surface contamination in a hospital facility's expressed human milk feed preparation area.

PubMed

Dippenaar, Ricky; Smith, Johan

2018-02-23

Expressed human milk (EHM) feed preparation areas represent a potential source of unintentional nosocomial infection. Daily disinfection of environmental surfaces remains an essential intervention to mitigate nosocomial infections. The inefficiency of conventional cleaning and disinfection contributes to an increased risk for the acquisition of multi-drug resistant pathogens. "Non touch" technologies such as the pulsed xenon ultraviolet (PX-UVD) light device have documented sustained reduction in surface bacterial colonization and reduced cross contamination. The impact of a PX-UVD on surface colony forming units per square centimeter (cfu/cm 2 ) in feed preparation areas was evaluated following its implementation as standard care. A quasi-experimental study was performed documenting bacterial colonization from 6 high risk feed preparation areas in a community care hospital in South Africa. Pre and post conventional cleaning neutralizing rinse swabs were collected fortnightly over a 16 week control period prior to the introduction of the PX-UVD and compared to a matching set of samples for the PX-UVD period. A 90% reduction in total surface bioburden was noted from the control period (544 cfu/cm 2 ) compared to the corresponding PX-UVD period (50 cfu/cm 2 ). Sub -analysis of both the Pre-clean Control: Pre-clean PX-UVD counts as well as the Post-clean Control: Post-clean PX-UVD counts noted significant improvements (p < 0.001). A statistically significant improvement was noted between pre-and post-cleaning total surface bioburden following exposure to the PX-UVD (p = 0.0004). The introduction of the PX-UVD was associated with a sustained reduction in the pre clean bioburden counts with a risk trend (per week) 0.19, (95% CI [0.056, 0.67], p = 0.01). The use of a PX-UVD as adjunct to standard cleaning protocols was associated with a significant decrease in surface bioburden. The study demonstrated the inefficiency of conventional cleaning. Persistence of potentially pathological species in both periods highlights current health sector challenges.

Viability and virulence of pneumolysin, pneumococcal surface protein A, and pneumolysin/pneumococcal surface protein A mutants in the ear.

PubMed

Schachern, Patricia A; Tsuprun, Vladimir; Goetz, Sarah; Cureoglu, Sebahattin; Juhn, Steven K; Briles, David E; Paparella, Michael M; Ferrieri, Patricia

2013-09-01

Understanding how pneumococcal proteins affect the pathology of the middle ear and inner ear is important for the development of new approaches to prevent otitis media and its complications. To determine the viability and virulence of Streptococcus pneumoniae mutants deficient in pneumolysin (Ply-) and pneumococcal surface protein A (PspA-) in the chinchilla middle ear. Bullae of chinchillas were inoculated bilaterally with wild-type (Wt), Ply-, PspA-, and Ply-/PspA- strains. Bacterial colony-forming units (CFUs) in middle ear effusions were counted at 48 hours. The CFUs of the PspA- group were also counted at 6 to 36 hours after inoculation. Temporal bone histopathological results were compared. Twenty-seven chinchillas in an academic research laboratory. Chinchilla middle ears were inoculated with S pneumoniae to produce sufficient volumes of effusions and noticeable histopathological changes in the ears. The CFU counts in the middle ear effusions and histopathological changes were compared to determine the effect of pneumococcal protein mutations on chinchilla ears. At 48 hours, CFUs in middle ears were increased for the Wt and Ply-/PspA- strains, but Ply- remained near inoculum level. No bacteria were detected in the PspA- group. The CFUs of PspA- decreased over time to a low level at 30 to 36 hours. In vitro, PspA- in Todd-Hewitt broth showed an increase in bacterial growth of 2 logs at 43 hours, indicating PspA- susceptibility to host defenses in vivo. The PspA- and Ply- groups had fewer pathologic findings than the Wt or Ply-/PspA- groups. Histopathological analysis showed significant differences in the number of bacteria in the scala tympani in the Wt group compared with the Ply-, PspA-, and Ply-/PspA- groups. The PspA- strain was the least virulent. The PspA- mutant was much less viable and less virulent in the ear than the Wt, Ply-, and Ply-/PspA- strains. There was no significant attenuation in the viability and virulence of the Ply-/PspA- mutant compared with the Wt or single mutants. The viability and virulence of pneumococcal mutants seemed to be protein and organ specific.

7 CFR 58.520 - Nondairy ingredients.

Code of Federal Regulations, 2010 CFR

2010-01-01

... grade quality and free from extraneous material. (b) Salt. Salt shall be free flowing, white refined... reasonably free of bacteria so as not to appreciably increase the bacterial count of the finished product...

7 CFR 58.520 - Nondairy ingredients.

Code of Federal Regulations, 2012 CFR

2012-01-01

... grade quality and free from extraneous material. (b) Salt. Salt shall be free flowing, white refined... reasonably free of bacteria so as not to appreciably increase the bacterial count of the finished product...

7 CFR 58.520 - Nondairy ingredients.

Code of Federal Regulations, 2013 CFR

2013-01-01

... grade quality and free from extraneous material. (b) Salt. Salt shall be free flowing, white refined... reasonably free of bacteria so as not to appreciably increase the bacterial count of the finished product...

7 CFR 58.520 - Nondairy ingredients.

Code of Federal Regulations, 2011 CFR

2011-01-01

... grade quality and free from extraneous material. (b) Salt. Salt shall be free flowing, white refined... reasonably free of bacteria so as not to appreciably increase the bacterial count of the finished product...

7 CFR 58.520 - Nondairy ingredients.

Code of Federal Regulations, 2014 CFR

2014-01-01

... grade quality and free from extraneous material. (b) Salt. Salt shall be free flowing, white refined... reasonably free of bacteria so as not to appreciably increase the bacterial count of the finished product...

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.

The effect of an acidic cleanser versus soap on the skin pH and micro-flora of adult patients: a non-randomised two group crossover study in an intensive care unit.

PubMed

Duncan, Christine N; Riley, Thomas V; Carson, Kerry C; Budgeon, Charley A; Siffleet, Joanne

2013-10-01

To test the effects of two different cleansing regimens on skin surface pH and micro-flora, in adult patients in the intensive care unit (ICU). Forty-three patients were recruited from a 23-bed tertiary medical/surgical ICU. The nineteen patients in Group One were washed using soap for daily hygiene care over a four week period. In Group 2, 24 patients were washing daily using an acidic liquid cleanser (pH 5.5) over a second four week period. Skin pH measurements and bacterial swabs were sampled daily from each for a maximum of ten days or until discharged from the ICU. Skin surface pH and quantitative skin cultures (colony forming units). Skin pH measurements were lower in patients washed with pH 5.5 cleanser than those washed with soap. This was statistically significant for both the forearm (p = 0.0068) and leg (p = 0.0015). The bacterial count was not statistically significantly different between the two groups. Both groups demonstrated that bacterial counts were significantly affected by the length of stay in ICU (p = 0.0032). This study demonstrated that the product used in routine skin care significantly affects the skin pH of ICU patients, but not the bacterial colonisation. Bacterial colonisation of the skin increases with length of stay. Crown Copyright © 2013. Published by Elsevier Ltd. All rights reserved.

Bacterial carriage on the fingernails of OR nurses.

PubMed

Wynd, C A; Samstag, D E; Lapp, A M

1994-11-01

This study provides statistically significant data that demonstrate that chipped fingernail polish or fingernail polish worn longer than four days fosters increased numbers of bacteria on the fingernails of OR nurses after surgical hand scrubs. There were no significant correlations between fingernail length and the numbers of bacterial colonies on the fingernails of the study groups tested after performing a standard surgical hand scrub. A convenience sample of 102 perioperative nurses with either freshly polished fingernails; chipped fingernail polish; or natural, polish-free fingernails participated. The data suggest OR nurses can wear fresh fingernail polish on healthy fingernails without risking increased bacterial counts.

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.

The salivary microbiome is altered in the presence of a high salivary glucose concentration

PubMed Central

Hartman, Mor-Li; Shi, Ping; Hasturk, Hatice; Yaskell, Tina; Vargas, Jorel; Song, Xiaoqing; Cugini, Maryann; Barake, Roula; Alsmadi, Osama; Al-Mutawa, Sabiha; Ariga, Jitendra; Soparkar, Pramod; Behbehani, Jawad; Behbehani, Kazem

2017-01-01

Background Type II diabetes (T2D) has been associated with changes in oral bacterial diversity and frequency. It is not known whether these changes are part of the etiology of T2D, or one of its effects. Methods We measured the glucose concentration, bacterial counts, and relative frequencies of 42 bacterial species in whole saliva samples from 8,173 Kuwaiti adolescents (mean age 10.00 ± 0.67 years) using DNA probe analysis. In addition, clinical data related to obesity, dental caries, and gingivitis were collected. Data were compared between adolescents with high salivary glucose (HSG; glucose concentration ≥ 1.0 mg/d, n = 175) and those with low salivary glucose (LSG, glucose concentration < 0.1 mg/dL n = 2,537). Results HSG was associated with dental caries and gingivitis in the study population. The overall salivary bacterial load in saliva decreased with increasing salivary glucose concentration. Under HSG conditions, the bacterial count for 35 (83%) of 42 species was significantly reduced, and relative bacterial frequencies in 27 species (64%) were altered, as compared with LSG conditions. These alterations were stronger predictors of high salivary glucose than measures of oral disease, obesity, sleep or fitness. Conclusions HSG was associated with a reduction in overall bacterial load and alterations to many relative bacterial frequencies in saliva when compared with LSG in samples from adolescents. We propose that hyperglycemia due to obesity and/or T2D results in HSG and subsequent acidification of the oral environment, leading to a generalized perturbation in the oral microbiome. This suggests a basis for the observation that hyperglycemia is associated with an increased risk of dental erosion, dental caries, and gingivitis. We conclude that HSG in adolescents may be predicted from salivary microbial diversity or frequency, and that the changes in the oral microbial composition seen in adolescents with developing metabolic disease may the consequence of hyperglycemia. PMID:28249034

Baicalin Improves Survival in a Murine Model of Polymicrobial Sepsis via Suppressing Inflammatory Response and Lymphocyte Apoptosis

PubMed Central

Zou, Yun; Bo, Lulong; Wang, Fei; Lou, Jingsheng; Fan, Xiaohua; Bao, Rui; Wu, Youping; Chen, Feng; Deng, Xiaoming; Li, Jinbao

2012-01-01

Background An imbalance between overwhelming inflammation and lymphocyte apoptosis is the main cause of high mortality in patients with sepsis. Baicalin, the main active ingredient of the Scutellaria root, exerts anti-inflammatory, anti-apoptotic, and even antibacterial properties in inflammatory and infectious diseases. However, the therapeutic effect of baicalin on polymicrobial sepsis remains unknown. Methodology/Principal Findings Polymicrobial sepsis was induced by cecal ligation and puncture (CLP) in C57BL/6 mice. Mice were infused with baicalin intraperitoneally at 1 h, 6 h and 12 h after CLP. Survival rates were assessed over the subsequent 8 days. Bacterial burdens in blood and peritoneal cavity were calculated to assess the bacterial clearance. Neutrophil count in peritoneal lavage fluid was also calculated. Injuries to the lung and liver were detected by hematoxylin and eosin staining. Levels of cytokines, including tumor necrosis factor (TNF)-alpha, interleukin (IL)-6, IL-10 and IL-17, in blood and peritoneum were measured by enzyme-linked immunosorbent assay. Adaptive immune function was assessed by apoptosis of lymphocytes in the thymus and counts of different cell types in the spleen. Baicalin significantly enhanced bacterial clearance and improved survival of septic mice. The number of neutrophils in peritoneal lavage fluid was reduced by baicalin. Less neutrophil infiltration of the lung and liver in baicalin-treated mice was associated with attenuated injuries to these organs. Baicalin significantly reduced the levels of proinflammatory cytokines but increased the level of anti-inflammatory cytokine in blood and peritoneum. Apoptosis of CD3+ T cell was inhibited in the thymus. The numbers of CD4+, CD8+ T lymphocytes and dendritic cells (DCs) were higher, while the number of CD4+CD25+ regulatory T cells was lower in the baicalin group compared with the CLP group. Conclusions/Significance Baicalin improves survival of mice with polymicrobial sepsis, and this may be attributed to its antibacterial property as well as its anti-inflammatory and anti-apoptotic effects. PMID:22590504

The evolution of a valved hepatoduodenal intestinal conduit.

PubMed

Kaufman, B H; Luck, S R; Raffensperger, J G

1981-06-01

Ascending cholangitis remains among the most serious complications following operations for biliary disorders. The bacterial count of refluxing intestinal contents can be reduced by using an enteric conduit from the biliary tract to the relatively sterile duodenum. A valvular conduit prohibits reflux of intestinal contents and permits unobstructed antegrade flow of bile. This can be created by intussuscepting approximately 1 cm of intestine in the midportion of the conduit. During the last 3 yr, valvular conduits were created in 11 patients. Seven of these children were treated for biliary atresia, and 4 had operations for choledochal duct cysts. There have been no deaths or morbidity resulting from the use of the valvular conduits. The postoperative courses in these patients indicate that the use of an intussusception valve may be beneficial in the prevention of ascending cholangitis.

The effect of carbon dioxide on the shelf life of ready-to-eat shredded chicken breast stored under refrigeration.

PubMed

Rodriguez, M B R; Junior, C A Conte; Carneiro, C S; Franco, R M; Mano, S B

2014-01-01

The objective of the present study was to determine the shelf life of ready-to-eat cooked chicken breast fillets (shredded) stored in atmospheres that were modified with different concentrations of CO2 and to establish a relationship between the concentration of this gas and bacterial growth. The samples were divided into 7 groups with different packaging conditions: aerobiosis, vacuum, and 10, 30, 50, 70, and 90% CO2 (with the remaining volume filled with N2). All of the samples were stored at 4 ± 2°C for 28 d. During this period, pH tests and counts of aerobic heterotrophic mesophyll bacteria (AHMB), aerobic heterotrophic psychotropic bacteria (AHPB), Enterobacteriaceae, and lactic acid bacteria (LAB) were performed, and the gas compositions of the packaging atmospheres were verified. The pH of the aerobic packages increased during storage. However, the other treatments resulted in the opposite trend, with the CO2 concentration decreasing over the first 24 h and then remaining constant until the end of experiment. A gradual increase in the AHMB, AHPB, Enterobacteriaceae, and LAB counts was observed during storage; this increase was faster in the meat that was packed under aerobiosis conditions than in the other treatments. The treatments with a CO2 concentration above 10% exhibited lower Enterobacteriaceae growth, whereas LAB growth was discrete in all of the treatments, independent of the CO2 concentration. The shelf life of the samples packed with 90% CO2 was 28 d. Based on the AHMB and AHPB counts, the shelf life was 3 times longer than for the samples packed under aerobiosis conditions (9 d). The increased package CO2 concentration caused a reduction in the growth rate of the examined bacteria (r = 0.99), and treatment with 90% CO2 appears promising as a method with which to increase the product's shelf life.

Iron overload in lower international prognostic scoring system risk patients with myelodysplastic syndrome receiving red blood cell transfusions: Relation to infections and possible benefit of iron chelation therapy.

PubMed

Wong, Colleen A C; Wong, Shannon A Y; Leitch, Heather A

2018-04-01

An increased incidence of infections and infectious mortality has been reported in myelodysplastic syndromes (MDS) patients receiving red blood cell (RBC) transfusions. We examined incidence of infections requiring antibiotics, antifungal or antiviral medications in transfused lower International Prognostic Scoring System (IPSS) risk MDS patients and whether this differed with iron chelation therapy (ICT). 138 transfused MDS patients were lower IPSS risk. 59 received ICT; median duration was 13 months. There was no significant difference between groups in neutrophil count at first RBC transfusion or first infection. Infections included: bacterial, n = 88; viral; fungal; and mycobacterial; n = 2 each. In ICT and non-ICT patients, respectively, infections were (number [%]): patients, 23 (40.0%) and 22 (27.8%); episodes (median [range]), 2 (1-6) and 2 (1-5); hospitalizations, 16 (27.1%) and 8 (10.1%); and deaths, 0 (0%) and 1 (1.3%), p = NS for all. Median overall survival (OS) from first RBC transfusion was superior in ICT patients, p = 0.01, and remained significant in a multivariate analysis (MVA), p = 0.003. Median time to first infection (TTI) was 27 and 7.8 months, respectively, p < 0.0001, and ICT remained significant for TTI in an MVA, p = 0.02, hazard ratio 0.3. For ICT patients with blast count <5%, TTI was significantly superior (p = 0.004). In this retrospective analysis, for lower IPSS risk MDS patients receiving RBC transfusions, though number and type of infections were similar between groups and despite similar neutrophil counts, time to first infection was significantly longer in ICT patients (p < 0.0001). These results should be confirmed in larger, prospective analyses. Copyright © 2018 Elsevier Ltd. All rights reserved.

Surface Bacterial-Spore Assay Using Tb3+/DPA Luminescence

NASA Technical Reports Server (NTRS)

Ponce, Adrian

2007-01-01

Equipment and a method for rapidly assaying solid surfaces for contamination by bacterial spores are undergoing development. The method would yield a total (nonviable plus viable) spore count of a surface within minutes and a viable-spore count in about one hour. In this method, spores would be collected from a surface by use of a transparent polymeric tape coated on one side with a polymeric adhesive that would be permeated with one or more reagent(s) for detection of spores by use of visible luminescence. The sticky side of the tape would be pressed against a surface to be assayed, then the tape with captured spores would be placed in a reader that illuminates the sample with ultraviolet light and counts the green luminescence spots under a microscope to quantify the number of bacterial spores per unit area. The visible luminescence spots seen through the microscope would be counted to determine the concentration of spores on the surface. This method is based on the chemical and physical principles of methods described in several prior NASA Tech Briefs articles, including Live/Dead Spore Assay Using DPA-Triggered Tb Luminescence (NPO-30444), Vol. 27, No. 3 (March 2003), page 7a. To recapitulate: The basic idea is to exploit the observations that (1) dipicolinic acid (DPA) is present naturally only in bacterial spores; and (2) when bound to Tb3+ ions, DPA triggers intense green luminescence of the ions under ultraviolet excitation; (3) DPA can be released from the viable spores by using L-alanine to make them germinate; and (4) by autoclaving, microwaving, or sonicating the sample, one can cause all the spores (non-viable as well as viable) to release their DPA. One candidate material for use as the adhesive in the present method is polydimethysiloxane (PDMS). In one variant of the method for obtaining counts of all (viable and nonviable) spores the PDMS would be doped with TbCl3. After collection of a sample, the spores immobilized on the sticky tape surface would be lysed by heating or microwaving to release their DPA. Tb3+ ions from the TbCl3 would become bound to the released DPA. The tape would then be irradiated with ultraviolet and examined as described above. In another variant of the method - for obtaining counts of viable spores only - the PDMS would be doped with L-alanine in addition to TbCl3. As now envisioned, a fully developed apparatus for implementing this method would include a pulsed source of ultraviolet light and a time-gated electronic camera to record the images seen through the microscope during a prescribed exposure interval at a prescribed short time after an ultraviolet pulse. As in the method of the second-mentioned prior article, the pulsing and time-gating would be used to discriminate between the longer-lived Tb3+/DPA luminescence and the shorter-lived background luminescence in the same wavelength range. In a time-gated image, the bright luminescence from bacterial spores could easily be seen against a dark background.

Accuracy of Automated Flow Cytometry-Based Leukocyte Counts To Rule Out Urinary Tract Infection in Febrile Children: a Prospective Cross-Sectional Study

PubMed Central

Duong, Hong Phuoc; Wissing, Karl Martin; Tram, Nathalie; Mascart, Georges; Lepage, Philippe

2016-01-01

Automated flow cytometry of urine remains an incompletely validated method to rule out urinary tract infection (UTI) in children. This cross-sectional analytical study was performed to compare the predictive values of flow cytometry and a dipstick test as initial diagnostic tests for UTI in febrile children and prospectively included 1,106 children (1,247 episodes). Urine culture was used as the gold standard test for diagnosing UTI. The performance of screening tests to diagnose UTI were established using receiver operating characteristic (ROC) analysis. Among these 1,247 febrile episodes, 221 UTIs were diagnosed (17.7% [95% confidence interval {CI}, 15.6 to 19.8%]). The area under the ROC curve for flow cytometry white blood cell (WBC) counts (0.99 [95% CI, 0.98 to 0.99]) was significantly superior to that for red blood cell (0.74 [95% CI, 0.70 to 0.78]) and bacterial counts (0.89 [95% CI, 0.87 to 0.92]) (P < 0.001). Urinary WBC counts also had a significantly higher area under the ROC curve than that of the leukocyte esterase (LE) dipstick (0.92 [95% CI, 0.90 to 0.94]), nitrite dipstick (0.83 [95% CI, 0.80 to 0.87]), or the combination of positive LE and/or nitrite dipstick (0.91 [95% CI, 0.89 to 0.93]) test (P < 0.001). The presence of ≥35 WBC/μl of urine was the best cutoff point, yielding both a high sensitivity (99.5% [95% CI, 99 to 100%]) and an acceptable specificity (80.6% [95% CI, 78 to 83%]). Using this cutoff point would have reduced the number of samples sent to the laboratory for culture by 67%. In conclusion, the determination of urinary WBC counts by flow cytometry provides optimal performance as an initial diagnostic test for UTI in febrile children. PMID:27682127

Bacterial dispersion in relation to operating room clothing.

PubMed Central

Whyte, W.; Vesley, D.; Hodgson, R.

1976-01-01

The effect of operating clothing on the dispersal of bacterial particles from the wearers was studied in a dispersal chamber. A comparison was made of six gowns as well as four types of trousers. The gowns were of three basic types, namely a conventional cotton type, disposable types made of non-woven fabric and those of the total-body exhaust system (Charnley type). The dispersal chamber could simulate conditions as expected both in down-flow unidirectional ultra-clean systems and in a conventional turbulent plenum-ventilated system. It was found that the disposable gowns would reduce the dispersal rate by about 30% in the simulated conventionally ventilated system and about 65% in the laminar flow system. The total-body exhaust system (Charnley) would reduce the count by 10-fold in the conventional ventilated system and by 66-fold in the laminar-flow system. The poor performance of the gowns in conventionally ventilated systems was caused by the dispersal of bacterial particles from underneath the gown (about 80%). This was not reduced by the disposable gown and only partially by the Charnley type. This small drop would be further decreased in a conventionally ventilated operating-room as only scrubbed staff would wear the gown. In order to overcome this poor performance in conventionally ventilated operating-rooms impervious trousers would be required. Four types were studied and it was demonstrated that those made either from Ventile or non-woven fabric would reduce the bacterial dispersion fourfold. As these tests had been carried out in an artificial environment checks were carried out in the unidirectional-flow operating-room during total-hip arthroplasty. This was done by comparing conventional cotton gowns with non-woven gowns and total-body exhaust gowns. The results showed good correlation between the operating room and the chamber with the non-woven fabric gown but the total-body exhaust system did not perform as well in the operating room (12-fold compared to 66-fold) the difference being possibly due to the contribution from the patient. However, as this comparison was that which would be most open to influence from other variables confidence could be placed on the chamber test results. Values were also obtained for the total number of bacterial particles dispersed by persons during a standard exercise wearing different clothing. This count was dependent on the clothing worn but a median count of between 1000 and 1500 bacterial particles/min. would be expected when conventional clothing was worn, with a range of between 300 and 19,000. This count could be reduced to about 100/min. if a total-body exhaust suit was worn (range 30-400). PMID:778258

Profile of selected bacterial counts and Salmonella prevalence on raw poultry in a poultry slaughter establishment.

PubMed

James, W O; Williams, W O; Prucha, J C; Johnston, R; Christensen, W

1992-01-01

The USDA Food Safety and Inspection Service determined populations of bacteria on poultry during processing at a slaughter plant in Puerto Rico in November and December 1987. The plant was selected because of its management's willingness to support important changes in equipment and processing procedures. The plant was representative of modern slaughter facilities. Eight-hundred samples were collected over 20 consecutive 8-hour days of operation from 5 sites in the processing plant. Results indicated that slaughter, dressing, and chilling practices significantly decreased the bacterial contamination on poultry carcasses, as determined by counts of aerobic bacteria, Enterobacteriaceae, and Escherichia coli. Salmonella was not enumerated; rather, it was determined to be present or absent by culturing almost the entire rinse. The prevalence of Salmonella in the study decreased during evisceration, then increased during immersion chilling.

Chair-side detection of Prevotella Intermedia in mature dental plaque by its fluorescence.

PubMed

Nomura, Yoshiaki; Takeuchi, Hiroaki; Okamoto, Masaaki; Sogabe, Kaoru; Okada, Ayako; Hanada, Nobuhiro

2017-06-01

Prevotella intermedia/nigrescens is one of the well-known pathogens causing periodontal diseases, and the red florescence excited by the visible blue light caused by the protoporphyrin IX in the bacterial cells could be useful for the chair-side detection. The aim of this study was to evaluated levels of periodontal pathogen, especially P. intermedia in clinical samples of red fluorescent dental plaque. Thirty two supra gingival plaque samples from six individuals were measured its fluorescence at 640nm wavelength excited by 409nm. Periodontopathic bacteria were counted by the Invader PLUS PCR assay. Co-relations the fluorescence intensity and bacterial counts were analyzed by Person's correlation coefficient and simple and multiple regression analysis. Positive and negative predictive values of the fluorescence intensities for with or without P. intermedia in supragingival plaque was calculated. When relative fluorescence unit (RFU) were logarithmic transformed, statistically significant linear relations between RFU and bacterial counts were obtained for P. intermedia, Porphyromonas gingivalis and Tannerella forsythia. By the multiple regression analysis, only P. intermedia had statistically significant co-relation with fluorescence intensities. All of the fluorescent dental plaque contained P. intermedia m. In contrast, 28% of non-fluorescent plaques contained P. intermedia. To check the fluorescence dental plaque in the oral cavity could be the simple chair-side screening of the mature dental plaque before examining the periodontal pathogens especially P. intermedia by the PCR method. Copyright © 2017 Elsevier B.V. All rights reserved.

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.

Intermediate-level disinfection with accelerated hydrogen peroxide prevents accumulation of bacteria in Versajet™ tubing during repeated daily debridement using simulated-use testing with an inoculated pork hock.

PubMed

Gawaziuk, J P; Alfa, M J; Olson, N; Logsetty, S

2014-05-01

This study assesses the feasibility of using the Versajet™ system (VJS) on an inoculated pork hock (PH) skin surface sequentially for 8 days with daily cleaning and intermediate-level disinfection (ILD). Daily, PHs were inoculated with bacteria suspended in artificial test soil (ATS). An ILD protocol with accelerated hydrogen peroxide (AHP, OxivirTB(®)) was employed to clean and disinfect the VJS between debridements. PH skin contains 6.1-6.8×10(6)cfu/cm(2) bacteria. Bacterial counts in the handpiece and discharge hoses immediately after debridement of the PHs, and before cleaning, increased throughout the study period (5.19-6.43log10cfu/mL). Cleaning with the ILD protocol was reduced bacterial counts on the VJS by 6-log. Protein, a surrogate marker of organic contamination, was also reduced post-cleaning and ILD. Compared to a maximum post-debridement level of protein (57.9 μg/mL) obtained before ILD, VJS protein levels dropped to 9.8 (handpiece) and 13.8 μg/mL (discharge hose). Disinfection of the handpiece and discharge hose after debridement with AHP resulted in a 6-log reduction in bacterial count and 4.2 fold reduction in protein. An ILD protocol with an AHP may be a feasible method for serial skin surface debridements with the VJS for up to eight days. Copyright © 2013 Elsevier Ltd and ISBI. All rights reserved.

Efficacy of disinfecting solutions in removing biofilms from polyvinyl chloride tracheostomy tubes.

PubMed

Silva, Rodrigo C; Carver, Ryan A; Ojano-Dirain, Carolyn P; Antonelli, Patrick J

2013-01-01

Bacterial biofilms are prevalent in pediatric tracheostomy tubes (TTs) and are not completely cleared by standard cleaning with gauze and household detergents. We aimed to examine the effectiveness of different disinfecting solutions to remove Staphylococcus aureus (SA) and Pseudomonas aerginosa (PA) biofilms from TTs. Prospective, controlled, in vitro microbiologic study. Uniform coupons obtained from polyvinyl chloride (PVC) pediatric TTs were briefly exposed to human plasma. The samples were incubated in growth media with either PA or SA for 7 days, and total bacterial growth was monitored by media turbidity. Five sets of 18 coupons each were exposed for 5 minutes to one of five different solutions: 2% aqueous chlorhexidine gluconate solution, 0.3% aqueous sodium hypochlorite, Polident denture cleanser, 3% hydrogen peroxide, or preservative-free phosphate-buffered saline (PBS) as a negative control. Biofilm presence was measured with bacterial counts, and surface integrity was assessed with scanning electron microscopy (SEM). All treatments significantly reduced mean SA counts (P = <.001). Sodium hypochlorite and chlorhexidine were more effective than peroxide and Polident. Chlorhexidine, sodium hypochlorite, and peroxide reduced PA counts (P = .001, .001, and .002, respectively), but Polident tabs had no significant effect. SEM revealed preserved TT surface integrity after exposure to all solutions. Disinfection with sodium hypochlorite or chlorhexidine solutions significantly reduces SA and PA biofilms on PVC TTs. Standard home care of reusable pediatric TTs may be improved by use of these readily available solutions. Copyright © 2012 The American Laryngological, Rhinological, and Otological Society, Inc.

Platelet transfusion therapy in sub-Saharan Africa: bacterial contamination, recipient characteristics and acute transfusion reactions

PubMed Central

Hume, Heather A.; Ddungu, Henry; Angom, Racheal; Baluku, Hannington; Kajumbula, Henry; Kyeyune-Byabazaire, Dorothy; Orem, Jackson; Ramirez-Arcos, Sandra; Tobian, Aaron A.R.

2017-01-01

Background Little data are available on bacterial contamination (BC) of platelet units or acute transfusion reactions to platelet transfusions (PT) in sub-Saharan Africa (SSA). Methods This prospective observational study evaluated the rate of BC of whole blood derived platelet units (WB-PU), the utility of performing Gram stains (GS) to prevent septic reactions, characteristics of patients receiving PT and the rate of acute reactions associated with PT at the Uganda Cancer Institute in Kampala, Uganda. An aliquot of each WB-PU studied was taken to perform GS and culture using the Bactec™ 9120 instrument. Study participants were monitored for reactions. Results 337 WB-PU were evaluated for BC, of which 323 units were transfused in 151 transfusion episodes to 50 patients. The frequency of BC ranged from 0.3%–2.1% (according to criteria used to define BC). The GS had high specificity (99.1%), but low sensitivity to detect units with BC. The median platelet count prior to PT was 10,900 (IQR 6,000–18,900) cells/μL. 78% of PT were given to patients with no bleeding. Acute reactions occurred in 11 transfusion episodes, involving 13 WB-PU, for a rate of 7.3% (95%CI=3.7–12.7%) per transfusion episode. All recipients of units with positive bacterial cultures were receiving antibiotics at the time of transfusion; none experienced a reaction. Conclusions The rate of BC observed in this study is lower than previously reported in SSA, but still remains a safety issue. As GS appears to be an ineffective screening tool, alternate methods should be explored to prevent transfusing bacterially-contaminated platelets in SSA. PMID:27079627

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.

The Proprotein Convertase Subtilisin/Kexin FurinA Regulates Zebrafish Host Response against Mycobacterium marinum

PubMed Central

Ojanen, Markus J. T.; Turpeinen, Hannu; Cordova, Zuzet M.; Hammarén, Milka M.; Harjula, Sanna-Kaisa E.; Parikka, Mataleena; Rämet, Mika

2015-01-01

Tuberculosis is a chronic bacterial disease with a complex pathogenesis. An effective immunity against Mycobacterium tuberculosis requires both the innate and adaptive immune responses, including proper T helper (Th) type 1 cell function. FURIN is a proprotein convertase subtilisin/kexin (PCSK) enzyme, which is highly expressed in Th1 type cells. FURIN expression in T cells is essential for maintaining peripheral immune tolerance, but its role in the innate immunity and infections has remained elusive. Here, we utilized Mycobacterium marinum infection models in zebrafish (Danio rerio) to investigate how furin regulates host responses against mycobacteria. In steady-state furinAtd204e/+ fish reduced furinA mRNA levels associated with low granulocyte counts and elevated Th cell transcription factor expressions. Silencing furin genes reduced the survival of M. marinum-infected zebrafish embryos. A mycobacterial infection upregulated furinA in adult zebrafish, and infected furinAtd204e/+ mutants exhibited a proinflammatory phenotype characterized by elevated tumor necrosis factor a (tnfa), lymphotoxin alpha (lta) and interleukin 17a/f3 (il17a/f3) expression levels. The enhanced innate immune response in the furinAtd204e/+ mutants correlated with a significantly decreased bacterial burden in a chronic M. marinum infection model. Our data show that upregulated furinA expression can serve as a marker for mycobacterial disease, since it inhibits early host responses and consequently promotes bacterial growth in a chronic infection. PMID:25624351

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.

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.

An evaluation of bacterial contamination of barriers used in periapical tissue regeneration: Part 1--Bacterial adherence.

PubMed

Sharma, Priya; Mickel, André K; Chogle, Sami; Sharma, Prem Nath; Han, Yiping W; Jones, Jefferson J

2008-02-01

To compare the adherence of Prevotella melaninogenica and Enterococcus faecalis to 3 guided tissue regeneration membranes: Atrisorb, Lambone, and OsseoQuest. It was hypothesized that OsseoQuest would show increased bacterial adherence compared to Lambone and Atrisorb. The barriers were suspended in trypticase soy broth containing an inoculum of either P melaninogenica or E faecalis. The samples were incubated under appropriate conditions for 6, 24, and 48 hours. Following incubation, each membrane was mixed in fresh media in a vortex machine to dislodge adherent bacteria. The vortexed media was quantitatively assessed using serial dilutions for viable cell count. E faecalis exhibited higher adherence compared to P melaninogenica with time. Of the membranes tested, Lambone displayed the least bacterial adherence. An analysis of the results indicated that bacterial adherence was time-dependent for all membranes. Membrane structure, chemical configuration, hydrophobicity, and bacterial cell surface structure were suggested as factors contributing to variance in bacterial adherence.

Bacillus cereus in free-stall bedding.

PubMed

Magnusson, M; Svensson, B; Kolstrup, C; Christiansson, A

2007-12-01

To increase the understanding of how different factors affect the bacterial growth in deep sawdust beds for dairy cattle, the microbiological status of Bacillus cereus and coliforms in deep sawdust-bedded free stalls was investigated over two 14-d periods on one farm. High counts of B. cereus and coliforms were found in the entire beds. On average, 4.1 log(10) B. cereus spores, 5.5 log(10) B. cereus, and 6.7 log(10) coliforms per gram of bedding could be found in the upper layers of the sawdust likely to be in contact with the cows' udders. The highest counts of B. cereus spores, B. cereus, and coliforms were found in the bedding before fresh bedding was added, and the lowest immediately afterwards. Different factors of importance for the growth of B. cereus in the bedding material were explored in laboratory tests. These were found to be the type of bedding, pH, and the type and availability of nutrients. Alternative bedding material such as peat and mixtures of peat and sawdust inhibited the bacterial growth of B. cereus. The extent of growth of B. cereus in the sawdust was increased in a dose-dependent manner by the availability of feces. Urine added to different bedding material raised the pH and also led to bacterial growth of B. cereus in the peat. In sawdust, a dry matter content greater than 70% was needed to lower the water activity to 0.95, which is needed to inhibit the growth of B. cereus. In an attempt to reduce the bacterial growth of B. cereus and coliforms in deep sawdust beds on the farm, the effect of giving bedding daily or a full replacement of the beds was studied. The spore count of B. cereus in the back part of the free stalls before fresh bedding was added was 0.9 log units lower in stalls given daily bedding than in stalls given bedding twice weekly. No effect on coliform counts was found. Replacement of the entire sawdust bedding had an effect for a short period, but by 1 to 2 mo after replacement, the counts of B. cereus spores in the beds had increased about 2 log units and were as high as they were before bed replacement. Therefore, free-stall management could, to a limited extent, reduce the content of B. cereus spores in the beds by daily bedding and entire bed replacement.

Impact of polymeric membrane breakage on drinking water quality and an online detection method of the breakage.

PubMed

Wu, Qilong; Zhang, Zhenghua; Cao, Guodong; Zhang, Xihui

2017-10-15

Polymeric membrane has been widely used for the treatment of drinking water in China, and the total treating capacity has reached up to 3.8 million m 3 /d. However, the membrane breakage found in the membrane modules in many water treatment plants resulted in an increase in turbidity and bacterial amount in the membrane permeate. In this study, a membrane module running for 3 years in a full-scale application was examined in terms of the breaking positions and the numbers of the broken fibers. It was found that most of the breaking positions were mainly on the outlet side of the module and that the distance from these points to the outlet was about 1/10-2/10 length of the membrane module. The lab-scale tests showed that the increase of the numbers of the breaking fibers in the membrane module (the breaking fibers were from 1 to 4 of 75 fibers) resulted in the increase in turbidity, particle count and the amount of total bacteria and coliform bacteria. Meanwhile, the water quality after the filtration with broken membrane fibers was similar to the quality of the raw water, which indicated that once the membrane fiber breakage occurred in the membrane module, the quality of drinking water after membrane filtration was significantly affected. Furthermore, the breaking position closer to the outlet side of the membrane module exposed much higher microbiological risk than those in the middle or near the bottom side. A pilot scale test was conducted by using a membrane module with 6600 fibers, and the effect of the membrane breakage (1-4 broken fibers) on water quality was also investigated. The results indicated that periodical backwashing caused drastic fluctuation of turbidity, particle count and the bacterial amount in the permeate water, which might be due to the washing force and self-blocking action inside the hollow fibers. Moreover, there is a good quantitative relationship (R 2 = 0.945) between particle count and the bacterial amount, which indicated that an online detection of particle count can be used to evaluate the bacterial risk. It was also suggested that the online detection of particle count after backwashing within 100 s would be a quick and precise method to identify any fiber breakage in time. These results are very important for the safety issue in the application of polymeric membrane to water treatment plants.

Comparison of a new inorganic membrane filter (Anopore) with a track-etched polycarbonate membrane filter (Nuclepore) for direct counting of bacteria.

PubMed Central

Jones, S E; Ditner, S A; Freeman, C; Whitaker, C J; Lock, M A

1989-01-01

Bacterial counts obtained by using a new Anopore inorganic membrane filter were 21 to 33% higher than those obtained by using a Nuclepore polycarbonate membrane filter. In addition, the inorganic filter had higher flow rates, permitting lower vacuum pressures to be used, while the intrinsically flat, rigid surface resulted in easier focusing and sharp definition of bacteria across the whole field of view. Images PMID:2655539

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

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.

Multistix 10 SG Leukocyte Esterage Dipstick Testing in Rapid Bedside Diagnosis of Spontaneous Bacterial Peritonitis: A Prospective Study.

PubMed

Jha, Ashish K; Kumawat, Dal C; Bolya, Yasvant K; Goenka, Mahesh K

2012-09-01

Spontaneous bacterial peritonitis (SBP) requires rapid diagnosis and the initiation of antibiotics. Diagnosis of SBP is usually based on cytobacteriological examination of ascitic fluid. These tests require good laboratory facilities and reporting time of few hours to 1-2 day. However, the 24 h laboratory facilities not widely available in country like India. We evaluated the diagnostic utility of reagent strip (Multistix 10 SG(®)) for rapid diagnosis of SBP. The study was prospectively carried out on patients of cirrhosis with ascites. Bedside leukocyte esterase reagent strip testing was performed on ascitic fluid. Cell count as determined by colorimetric scale of reagent strip was compared with counting chamber method. Sensitivity, specificity, positive predictive value, negative predictive value, and accuracy were calculated. Out of 100 patients with cirrhotic ascites, [72 males: 28 female; mean age 44.34 (SD 13.03) years] 18 patients were diagnosed to have SBP by counting chamber method as compared to 14 patients detected to have SBP by reagent strip test ≥++ positive. The sensitivity, specificity, positive predictive value, negative predictive value and accuracy of reagent strip ≥++ positive were 77.77%, 95.12%, 77.77%, 95.12% and 92% respectively compared to counting chamber method. Reagent strip to diagnose SBP is very specific but less sensitive as compared to counting chamber method. This can be performed rapidly, easily and efficiently even in remote area of developing countries. This bedside test could be a useful tool for the diagnosis of SBP in country like India.

Predictors of bacterial pneumonia in Evaluation of Subcutaneous Interleukin-2 in a Randomized International Trial (ESPRIT).

PubMed

Pett, S L; Carey, C; Lin, E; Wentworth, D; Lazovski, J; Miró, J M; Gordin, F; Angus, B; Rodriguez-Barradas, M; Rubio, R; Tambussi, G; Cooper, D A; Emery, S

2011-04-01

Bacterial pneumonia still contributes to morbidity/mortality in HIV infection despite effective combination antiretroviral therapy (cART). Evaluation of Subcutaneous Interleukin-2 in a Randomized International Trial (ESPRIT), a trial of intermittent recombinant interleukin-2 (rIL-2) with cART vs. cART alone (control arm) in HIV-infected adults with CD4 counts ≥300cells/μL, offered the opportunity to explore associations between bacterial pneumonia and rIL-2, a cytokine that increases the risk of some bacterial infections. Baseline and time-updated factors associated with first-episode pneumonia on study were analysed using multivariate proportional hazards regression models. Information on smoking/pneumococcal vaccination history was not collected. IL-2 cycling was most intense in years 1-2. Over ≈7 years, 93 IL-2 [rate 0.67/100 person-years (PY)] and 86 control (rate 0.63/100 PY) patients experienced a pneumonia event [hazard ratio (HR) 1.06; 95% confidence interval (CI) 0.79, 1.42; P=0.68]. Median CD4 counts prior to pneumonia were 570cells/μL (IL-2 arm) and 463cells/μL (control arm). Baseline risks for bacterial pneumonia included older age, injecting drug use, detectable HIV viral load (VL) and previous recurrent pneumonia; Asian ethnicity was associated with decreased risk. Higher proximal VL (HR for 1 log(10) higher VL 1.28; 95% CI 1.11, 1.47; P<0.001) was associated with increased risk; higher CD4 count prior to the event (HR per 100 cells/μL higher 0.94; 95% CI 0.89, 1.0; P=0.04) decreased risk. Compared with controls, the hazard for a pneumonia event was higher if rIL-2 was received <180 days previously (HR 1.66; 95% CI 1.07, 2.60; P=0.02) vs.≥180 days previously (HR 0.98; 95% CI 0.70, 1.37; P=0.9). Compared with the control group, pneumonia risk in the IL-2 arm decreased over time, with HRs of 1.41, 1.71, 1.16, 0.62 and 0.84 in years 1, 2, 3-4, 5-6 and 7, respectively. Bacterial pneumonia rates in cART-treated adults with moderate immunodeficiency are high. The mechanism of the association between bacterial pneumonia and recent IL-2 receipt and/or detectable HIV viraemia warrants further exploration. © 2010 British HIV Association.

A Note on the Flora and Fauna in the Rumen of Steers Fed a Feedlot Bloat-provoking Ration and the Effect of Penicillin

PubMed Central

Bryant, M. P.; Robinson, I. M.; Lindahl, Ivan L.

1961-01-01

A study was made of the predominant culturable bacteria and ciliate protozoa present in the rumen of two steers that were regularly bloating on a pelleted ratio containing 22% alfalfa meal, 16% soybean oil meal, 61% barley, and 1% common salt. The ruminal microorganisms in the two animals differed as indicated by a high total culture count of bacteria, an almost complete absence of ciliate protozoa, a low pH, and a difference in the proportions of presumptively identified predominant bacterial groups in one animal (steer 26) as compared with the other (steer 32). The first exposure of the animals to procaine penicillin (75 or 150 mg per day on 2 successive days) resulted in an abnormal ruminal flora 31 hr after the first treatment as indicated by drastic drops in total and cellulolytic bacterial counts and a change in the proportions of predominant bacterial groups. The animals refused feed for 32 to 48 hr after the first treatment. After feed consumption resumed, further treatment with 75 mg penicillin on 4 successive days did not appear to greatly alter the flora and did not result in feed refusal in animal 32. In animal 26, amounts of penicillin progressing from 50 to 200 mg per day did not result in feed refusals and observations on rumen ingesta samples during this period indicated a decrease in total bacterial count, a great increase in numbers of ciliate protozoa, a higher pH, and a change in the proportions of predominant bacterial groups so that the ruminal picture was much more similar to that of animal 32 than to its own during the pre-penicillin period. Bloat was not relieved except during the period of feed refusal. The results indicate that the ruminal flora rapidly adapts to penicillin and that bloat of the feedlot type can occur in animals with widely differing numbers and kinds of bacteria and protozoa. Feedlot bloat does not appear to be correlated with the occurrence or numbers of any of the individual predominant groups of bacteria cultured. PMID:13874139

A note on the flora and fauna in the rumen of steers fed a feedlot bloat-provoking ration and the effect of penicillin.

PubMed

BRYANT, M P; ROBINSON, I M; LINDAHL, I L

1961-11-01

A study was made of the predominant culturable bacteria and ciliate protozoa present in the rumen of two steers that were regularly bloating on a pelleted ratio containing 22% alfalfa meal, 16% soybean oil meal, 61% barley, and 1% common salt. The ruminal microorganisms in the two animals differed as indicated by a high total culture count of bacteria, an almost complete absence of ciliate protozoa, a low pH, and a difference in the proportions of presumptively identified predominant bacterial groups in one animal (steer 26) as compared with the other (steer 32). The first exposure of the animals to procaine penicillin (75 or 150 mg per day on 2 successive days) resulted in an abnormal ruminal flora 31 hr after the first treatment as indicated by drastic drops in total and cellulolytic bacterial counts and a change in the proportions of predominant bacterial groups. The animals refused feed for 32 to 48 hr after the first treatment. After feed consumption resumed, further treatment with 75 mg penicillin on 4 successive days did not appear to greatly alter the flora and did not result in feed refusal in animal 32. In animal 26, amounts of penicillin progressing from 50 to 200 mg per day did not result in feed refusals and observations on rumen ingesta samples during this period indicated a decrease in total bacterial count, a great increase in numbers of ciliate protozoa, a higher pH, and a change in the proportions of predominant bacterial groups so that the ruminal picture was much more similar to that of animal 32 than to its own during the pre-penicillin period. Bloat was not relieved except during the period of feed refusal. The results indicate that the ruminal flora rapidly adapts to penicillin and that bloat of the feedlot type can occur in animals with widely differing numbers and kinds of bacteria and protozoa. Feedlot bloat does not appear to be correlated with the occurrence or numbers of any of the individual predominant groups of bacteria cultured.

Vitamin E Phosphate Coating Stimulates Bone Deposition in Implant-related Infections in a Rat Model.

PubMed

Lovati, Arianna B; Bottagisio, Marta; Maraldi, Susanna; Violatto, Martina B; Bortolin, Monica; De Vecchi, Elena; Bigini, Paolo; Drago, Lorenzo; Romanò, Carlo L

2018-06-01

Implant-related infections are associated with impaired bone healing and osseointegration. In vitro antiadhesive and antibacterial properties and in vivo antiinflammatory effects protecting against bone loss of various formulations of vitamin E have been demonstrated in animal models. However, to the best of our knowledge, no in vivo studies have demonstrated the synergistic activity of vitamin E in preventing bacterial adhesion to orthopaedic implants, thus supporting the bone-implant integration. The purpose of this study was to test whether a vitamin E phosphate coating on titanium implants may be able to reduce (1) the bacterial colonization of prosthetic implants and (2) bone resorption and osteomyelitis in a rat model of Staphylococcus aureus-induced implant-related infection. Twelve rats were bilaterally injected in the femurs with S aureus UAMS-1-Xen40 and implanted with uncoated or vitamin E phosphate-coated titanium Kirschner wires without local or systemic antibiotic prophylaxis. Eight rats represented the uninfected control group. A few hours after surgery, two control and three infected animals died as a result of unexpected complications. With the remaining rats, we assessed the presence of bacterial contamination with qualitative bioluminescence imaging and Gram-positive staining and with quantitative bacterial count. Bone changes in terms of resorption and osteomyelitis were quantitatively analyzed through micro-CT (bone mineral density) and semiquantitatively through histologic scoring systems. Six weeks after implantation, we found only a mild decrease in bacterial count in coated versus uncoated implants (Ti versus controls: mean difference [MD], -3.705; 95% confidence interval [CI], -4.416 to -2.994; p < 0.001; TiVE versus controls: MD, -3.063; 95% CI, -3.672 to -2.454; p < 0.001), whereas micro-CT analysis showed a higher bone mineral density at the knee and femoral metaphysis in the vitamin E-treated group compared with uncoated implants (knee joint: MD, -11.88; 95% CI, -16.100 to -7.664; p < 0.001 and femoral metaphysis: MD, -19.87; 95% CI, -28.82 to -10.93; p < 0.001). We found decreased osteonecrosis (difference between medians, 1.5; 95% CI, 1-2; p < 0.002) in the infected group receiving the vitamin E-coated nails compared with the uncoated nails. These preliminary findings indicate that vitamin E phosphate implant coatings can exert a protective effect on bone deposition in a highly contaminated animal model of implant-related infection. The use of vitamin E coatings may open new perspectives for developing coatings that can limit septic loosening of infected implants with bacterial contamination. However, a deeper insight into the mechanism of action and the local release of vitamin E as a coating for orthopaedic implants is required to be used in clinics in the near future. Although this study cannot support the antimicrobial properties of vitamin E, promising results were obtained for bone-implant osseointegration. These preliminary results will require further in vivo investigations to optimize the host response in the presence of antibiotic prophylaxis.

Differential bacterial load on components of total knee prosthesis in patients with prosthetic joint infection.

PubMed

Holinka, Johannes; Pilz, Magdalena; Hirschl, Alexander M; Graninger, Wolfgang; Windhager, Reinhard; Presterl, Elisabeth

2012-10-01

The purpose of our study was to evaluate and quantify the bacterial adherence on different components of total knee prosthesis with the sonication culture method. Explanted components of all patients with presumptive prosthetic or implant infection were treated by sonication separately in sterile containers to dislodge the adherent bacteria from the surfaces and cultured. The bacterial load of the different knee components (femur, tibia, PE-inlay and patella) was evaluated by counting of colony-forming units (CFU) dislodged from the components surfaces using the sonication culture method. Overall, 27 patients had positive sonication cultures of explanted total knee prostheses. Microorganisms were detected from 88 of 100 explanted components. Twenty femoral components were culture positive and 7 negative, 23 tibial components as well as 23 polyethylene (PE) platforms had positive microorganism detection from the surface. Staphylococcus epidermidis adhered to the highest number of components whereas Staphylococcus aureus yielded the highest load of CFU in the sonication cultures. Although not significant, PE-inlays and tibial components were most often affected. The highest CFU count was detected in polyethylene components. The sonication culture method is a reliable method to detect bacteria from the components. Additionally, the results demonstrate that bacterial adherence is not affecting a single component of knee prosthesis only. Thus, in septic revision surgery partial prosthetic exchange or exchange of single polyethylene components alone may be not sufficient.

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.

Comparison of the cytotoxic effect of polystyrene latex nanoparticles on planktonic cells and bacterial biofilms

NASA Astrophysics Data System (ADS)

Nomura, Toshiyuki; Fujisawa, Eri; Itoh, Shikibu; Konishi, Yasuhiro

2016-06-01

The cytotoxic effect of positively charged polystyrene latex nanoparticles (PSL NPs) was compared between planktonic bacterial cells and bacterial biofilms using confocal laser scanning microscopy, atomic force microscopy, and a colony counting method. Pseudomonas fluorescens, which is commonly used in biofilm studies, was employed as the model bacteria. We found that the negatively charged bacterial surface of the planktonic cells was almost completely covered with positively charged PSL NPs, leading to cell death, as indicated by the NP concentration being greater than that required to achieve single layer coverage. In addition, the relationship between surface coverage and cell viability of P. fluorescens cells correlated well with the findings in other bacterial cells ( Escherichia coli and Lactococcus lactis). However, most of the bacterial cells that formed the biofilm were viable despite the positively charged PSL NPs being highly toxic to planktonic bacterial cells. This indicated that bacterial cells embedded in the biofilm were protected by self-produced extracellular polymeric substances (EPS) that provide resistance to antibacterial agents. In conclusion, mature biofilms covered with EPS exhibit resistance to NP toxicity as well as antibacterial agents.

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.

Is it possible to sanitize athletes' shoes?

PubMed

Messina, Gabriele; Burgassi, Sandra; Russo, Carmela; Ceriale, Emma; Quercioli, Cecilia; Meniconi, Cosetta

2015-02-01

Footwear should be designed to avoid trauma and injury to the skin of the feet that can favor bacterial and fungal infections. Procedures and substances for sanitizing the interior of shoes are uncommon but are important aspects of primary prevention against foot infections and unpleasant odor. To evaluate the efficacy of a sanitizing technique for reducing bacterial and fungal contamination of footwear. Crossover study. Mens Sana basketball team. Twenty-seven male athletes and 4 coaches (62 shoes). The experimental protocol required a first sample (swab), 1/shoe, at time 0 from inside the shoes of all athletes before the sanitizing technique began and a second sample at time 1, after about 4 weeks, April 2012 to May 2012, of daily use of the sanitizing technique. The differences before and after use of the sanitizing technique for total bacterial count at 36 °C and 22 °C for Staphylococcus spp, yeasts, molds, Enterococcus spp, Pseudomonas spp, Escherichia coli , and total coliform bacteria were evaluated. Before use of the sanitizing technique, the total bacterial counts at 36 °C and 22 °C and for Staphylococcus spp were greater by a factor of 5.8 (95% confidence interval [CI] = 3.42, 9.84), 5.84 (95% CI = 3.45, 9.78), and 4.78 (95% CI = 2.84, 8.03), respectively. All the other comparisons showed a reduction in microbial loads, whereas E coli and coliforms were no longer detected. No statistically significant decrease in yeasts (P = .0841) or molds (P = .6913) was recorded probably because of low contamination. The sanitizing technique significantly reduced the bacterial presence in athletes' shoes.

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.

Inclusion of detergent in a cleaning regime and effect on microbial load in livestock housing.

PubMed

Hancox, L R; Le Bon, M; Dodd, C E R; Mellits, K H

Determining effective cleaning and disinfection regimes of livestock housing is vital to improving the health of resident animals and reducing zoonotic disease. A cleaning regime consisting of scraping, soaking with or without detergent (treatment and control), pressure washing, disinfection and natural drying was applied to multiple pig pens. After each cleaning stage, samples were taken from different materials and enumerated for total aerobic count (TAC) and Enterobacteriaceae (ENT). Soaking with detergent (Blast-Off, Biolink) caused significantly greater reductions of TAC and ENT on metal, and TAC on concrete, compared with control. Disinfection effect (Virkon S, DuPont) was not significantly associated with prior detergent treatment. Disinfection significantly reduced TAC and ENT on concrete and stock board but not on metal. Twenty-four hours after disinfection TAC and ENT on metal and stock board were significantly reduced, but no significant reductions occurred in the subsequent 96 hours. Counts on concrete did not significantly reduce during the entire drying period (120 hours). Detergent and disinfectant have varying bactericidal effects according to the surface and bacterial target; however, both can significantly reduce microbial numbers so should be used during cleaning, with a minimum drying period of 24 hours, to lower bacterial counts effectively.

Evaluation on the effects of 0.1% Peumus boldus leaf and Spiraea ulmaria plant extract combination on bacterial colonization in canine atopic dermatitis: A preliminary randomized, placebo controlled, double-blinded study.

PubMed

Santoro, Domenico; Bohannon, Mary; Ahrens, Kim; Navarro, Christelle; Gatto, Hugues; Marsella, Rosanna

2018-06-01

Defective skin barrier characterize canine atopic dermatitis (AD). Pyoderma is the most common complication. Herbal compounds have been suggested as alternatives to control bacterial colonization for their effect on natural antimicrobial peptides (AMPs). This study evaluated the effects of 0.1% Peumus boldus leaf and Spiraea ulmaria plant extract combination on clinical signs, bacterial colonization and AMPs secretion in atopic dogs compared to placebo. Twenty privately-owned atopic dogs were randomly divided in 2 groups (treatment: n = 10; placebo: n = 10) and their abdomen was sprayed every 24 h for 4 weeks. Total and inguinal clinical scores (CADESI-03), manual bacterial count, and skin washes for AMPs (cBD3-like and cCath) were performed on days 0, 14 and 28. AMPs were detected using in-house, previously-validated, canine-specific ELISAs. Data were statistically analyzed and a p < 0.05 was considered significant. Clinical scores and AMPs secretion did not differ significantly between the two groups at any time point. A significant reduction of the clinical scores was seen in the placebo group at 14 and 28 days (p < 0.04). On days 14 and 28, a reduction in the bacterial count was seen in the treated group compared with placebo (p < 0.009 and p = 0.04, respectively). Compared to baseline, a reduction in Staphylococcus spp. was seen in the treated group after 14 days of treatment (p < 0.03). These results show the efficacy of this plant extract combination against bacterial colonization, suggesting its potential usefulness in preventing bacterial infection in atopic dogs. The influence of this compound on AMPs secretion or other mechanisms should be further evaluated. Copyright © 2018 The Authors. Published by Elsevier Ltd.. All rights reserved.

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.

Assessing the cleanliness of surfaces: Innovative molecular approaches vs. standard spore assays

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

Cooper, M.; Duc, M.T. La; Probst, A.

2011-04-01

A bacterial spore assay and a molecular DNA microarray method were compared for their ability to assess relative cleanliness in the context of bacterial abundance and diversity on spacecraft surfaces. Colony counts derived from the NASA standard spore assay were extremely low for spacecraft surfaces. However, the PhyloChip generation 3 (G3) DNA microarray resolved the genetic signatures of a highly diverse suite of microorganisms in the very same sample set. Samples completely devoid of cultivable spores were shown to harbor the DNA of more than 100 distinct microbial phylotypes. Furthermore, samples with higher numbers of cultivable spores did not necessarilymore » give rise to a greater microbial diversity upon analysis with the DNA microarray. The findings of this study clearly demonstrated that there is not a statistically significant correlation between the cultivable spore counts obtained from a sample and the degree of bacterial diversity present. Based on these results, it can be stated that validated state-of-the-art molecular techniques, such as DNA microarrays, can be utilized in parallel with classical culture-based methods to further describe the cleanliness of spacecraft surfaces.« less

Microbial Characterization of Qatari Barchan Sand Dunes

PubMed Central

Chatziefthimiou, Aspassia D.; Nguyen, Hanh; Richer, Renee; Louge, Michel; Sultan, Ali A.; Schloss, Patrick; Hay, Anthony G.

2016-01-01

This study represents the first characterization of sand microbiota in migrating barchan sand dunes. Bacterial communities were studied through direct counts and cultivation, as well as 16S rRNA gene and metagenomic sequence analysis to gain an understanding of microbial abundance, diversity, and potential metabolic capabilities. Direct on-grain cell counts gave an average of 5.3 ± 0.4 x 105 cells g-1 of sand. Cultured isolates (N = 64) selected for 16S rRNA gene sequencing belonged to the phyla Actinobacteria (58%), Firmicutes (27%) and Proteobacteria (15%). Deep-sequencing of 16S rRNA gene amplicons from 18 dunes demonstrated a high relative abundance of Proteobacteria, particularly enteric bacteria, and a dune-specific-pattern of bacterial community composition that correlated with dune size. Shotgun metagenome sequences of two representative dunes were analyzed and found to have similar relative bacterial abundance, though the relative abundances of eukaryotic, viral and enterobacterial sequences were greater in sand from the dune closer to a camel-pen. Functional analysis revealed patterns similar to those observed in desert soils; however, the increased relative abundance of genes encoding sporulation and dormancy are consistent with the dune microbiome being well-adapted to the exceptionally hyper-arid Qatari desert. PMID:27655399

Evaluation of the automated urine particle analyzer UF-1000i screening for urinary tract infection in nonpregnant women.

PubMed

Dai, Qingkai; Jiang, Yongmei; Shi, Hua; Zhou, Wei; Zhou, Shengjie; Yang, Hui

2014-01-01

Urinary tract infection (UTI) is a widespread disease in women. Urine culture is still the "gold standard" diagnostic test for UTI, but most of them are negative. To reduce unnecessary culture, we evaluated the automated urine particle analyzer UF-1000i screening for UTI in nonpregnant women. The urine specimens submitted to our laboratory were submitted for culture and tested by the Sysmex UF-1000i. Bacteria and white blood cell (WBC) counts were compared to standard urine culture results to assess the best cutoff values. In this study, 272 urine samples were included, of which 98 (36.0%) were culture positive with a bacterial cutoff value of 10 x 10(5) CFU/mL. A combination of bacterial (> 95/microL) and/or WBC count (> 24/microL) provided the best screening for UTI, with a sensitivity of 0.99 and a specificity of 0.82 compared with the urine culture. Sysmex UF-1000i could be used as a screening test for UTI in nonpregnant women. According to the distribution and range of the bacterial scattergram, we could primarily identify and differentiate between Gram-negative and Gram-positive bacteria.

Dormant Cells of Staphylococcus aureus Are Resuscitated by Spent Culture Supernatant

PubMed Central

Pascoe, Ben; Dams, Lucy; Wilkinson, Tom S.; Harris, Llinos G.; Bodger, Owen; Mack, Dietrich; Davies, Angharad P.

2014-01-01

We describe the first in vitro model of dormancy in Staphylococcus aureus, showing that cells are generated which can be resuscitated by addition of spent medium supernatant taken from cultures of the same organism. Over 30 days, culturable counts in dormant cultures of S. aureus SH1000 fell from 106–107 cfu/ml to <10 cfu/ml as measured by the Most Probable Number method in liquid culture, while total counts as determined by microscopy, and supported by data from RT-qPCR, remained around 106–107 cells/ml. Supplementing cultures with 25–50% spent medium resulted in a >600-fold increase in bacterial growth. Resuscitation was a specific effect, greatly reduced by boiling or addition of trypsin to the spent supernatant. Supernatant also effected a reduction in lag phase of dormant cultures. SEM demonstrated the presence of small coccoid cells in dormant cultures. The results are similar to those seen with resuscitation promoting factors (Rpfs) in actinobacteria. This is the first time resuscitation has been demonstrated in Staphylococcus aureus, which is an important human pathogen. A better understanding of control and reactivation of dormant cells could lead to major improvements in managing staphylococcal infections; resuscitation could be an important step in restoring susceptibility to antibiotic treatment. PMID:24523858

Coliform bacteria removal from sewage in constructed wetlands planted with Mentha aquatica.

PubMed

Avelar, Fabiana F; de Matos, Antonio T; de Matos, Mateus P; Borges, Alisson C

2014-08-01

The present study evaluated the performance of the species Mentha aquatica in constructed wetlands of horizontal subsurface flow (CW-HSSF) with regard to the removal of coliforms bacteria in an effluent from the primary treatment of sewage as well as to obtain adjustment parameters of the bacterial decay kinetic model along the length of the CW-HSSF. Therefore, four CW-HSSFs measuring 24.0 m x 1.0 m x 0.35 m were built and filled with number 0 gravel as the support medium to a height of 0.20m. Two of the CW-HSSFs were planted with the species M. aquatica, while the other two remained uncultivated. Cultivation of M. aquatica in CW-HSSF resulted in total coliforms (TC) and Escherichia coli (EC) removals from 0.9 to 1.3 log units greater than those obtained in the uncultivated experimental plots, for the hydraulic retention times (HRTs) of 4.5 and 6.0 days. For HRT ranged from 1.5 to 6.0 days, the highest removal efficiencies in counts of TC and EC were obtained when using longer HRT. The mathematical models evaluated showed good fit to average counts of TC and EC highlighting the modified first-order kinetic model with the inclusion of the power parameter in the HRT variable.

Contamination pathways of spore-forming bacteria in a vegetable cannery.

PubMed

Durand, Loïc; Planchon, Stella; Guinebretiere, Marie-Hélène; André, Stéphane; Carlin, Frédéric; Remize, Fabienne

2015-06-02

Spoilage of low-acid canned food during prolonged storage at high temperatures is caused by heat resistant thermophilic spores of strict or facultative bacteria. Here, we performed a bacterial survey over two consecutive years on the processing line of a French company manufacturing canned mixed green peas and carrots. In total, 341 samples were collected, including raw vegetables, green peas and carrots at different steps of processing, cover brine, and process environment samples. Thermophilic and highly-heat-resistant thermophilic spores growing anaerobically were counted. During vegetable preparation, anaerobic spore counts were significantly decreased, and tended to remain unchanged further downstream in the process. Large variation of spore levels in products immediately before the sterilization process could be explained by occasionally high spore levels on surfaces and in debris of vegetable combined with long residence times in conditions suitable for growth and sporulation. Vegetable processing was also associated with an increase in the prevalence of highly-heat-resistant species, probably due to cross-contamination of peas via blanching water. Geobacillus stearothermophilus M13-PCR genotypic profiling on 112 isolates determined 23 profile-types and confirmed process-driven cross-contamination. Taken together, these findings clarify the scheme of contamination pathway by thermophilic spore-forming bacteria in a vegetable cannery. Copyright © 2015 Elsevier B.V. All rights reserved.

Comparison of two fluorescent antibody techniques (FATS) for detection and quantification of Renibacterium salmoninarum in coelomic fluid of spawning chinook salmon Oncorhynchus tshawytscha

USGS Publications Warehouse

Elliott, D.G.; McKibben, C.L.

1997-01-01

Two versions of the fluorescent antibody technique (FAT) were compared for detection and quantification of Renibacterium salmoninarum in coelomic fluid samples from naturally infected spawning chinook salmon Oncorhynchus tshawytscha. For the membrane filtration-FAT (MF-FAT), trypsin-treated samples were passed through 0.2 ??m polycarbonate filters to concentrate bacteria for direct enumeration by immunofluorescence microscopy. For the smear-FAT (S-FAT), samples were centrifuged at 8800 x g for 10 min and the pelleted material was smeared on slides for immunofluorescence staining Detected prevalences of Renibacterium salmoninarum were 1.8 to 3.4 times higher by the MF-FAT than by the S-FAT: differences were significant at p ??? 0.0002. The S-FAT consistently detected R. salmoninarum only in samples with calculated bacterial concentrations ??? 2.4 x 103 cells ml-1 by MF-FAT testing. Increasing the area examined on a filter or slide from 50 to 100 microscope fields at 1000x magnification resulted in the detection of a maximum of 4% additional positive samples by the MF-FAT and 7% additional positive samples by the S-FAT. In individual samples for which bacterial counts were obtained by both the MF-FAT and the S-FAT, the counts averaged from 47 times (??30 SD) to 175 times (??165 SD) higher by the MF-FAT. Centrifugation of samples at 10000 x g for 10 min resulted in a 4-fold increase in mean bacterial counts by the S-FAT compared with a 10-min centrifugation at 2000 x g, but the highest calculated bacterial concentration obtained by S-FAT testing was more than 6-fold lower than that obtained for the same sample by MF-FAT testing. Because of its greater sensitivity, the MF-FAT is preferable to the S-FAT for use in critical situations requiring the detection of low numbers of R. salmoninarum.

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.

The antimicrobial effectiveness of 25% propolis extract in root canal irrigation of primary teeth.

PubMed

Verma, Manjesh Kumar; Pandey, Ramesh Kumar; Khanna, Richa; Agarwal, Jyotsna

2014-01-01

The choice of irrigating solution used in root canals of primary teeth is complicated by their complex morphology and paucity of associated literature. Propolis is a natural product that has gained interest in this context due to its antibacterial effectiveness against several endodontic pathogens. The present study was undertaken to assess the potential of water-soluble 25% propolis extract against microorganisms present in root canals of primary teeth during endodontic procedures. The child patients in the age group of 4-7 years with radiographic evidence of carious pulp exposure were included in the study. Definitive selection was done after gaining access into the pulp chamber and root canals of the selected teeth. The clinical and radiographic evidence of pathosis was ruled out for inclusion in the study. The selected teeth were divided into two groups randomly. In Group A 0.9% isotonic saline and in Group B 25% extract water-soluble propolis were used as irrigating solution, respectively. The bacterial samples were collected both pre- and post-irrigation and were transferred for microbial assay. STAISTISTICAL ANALYSIS: Wilcoxon matched signed rank test was used to compare the pre-and post-irrigation bacterial counts. Mann-Whitney test was used to compare the mean change (pre-post) in bacterial colony counts of groups in the study. Antimicrobial effectiveness of 25% water-soluble extract of propolis in the root canals of primary teeth was confirmed in the present study. The reduction in the mean bacterial colony counts of all the isolated bacteria was noticed higher in Group B than Group A. The results of the present study have confirmed that the antibacterial effectiveness of water-soluble extract of propolis in the root canals of primary teeth in vivo. Considering the low toxicity concerns and antibacterial effectiveness, water-soluble extract of 25% propolis can be advocated as a root canal irrigant in endodontic treatment of primary teeth.

[Predictive factors of contamination in a blood culture with bacterial growth in an Emergency Department].

PubMed

Hernández-Bou, S; Trenchs Sainz de la Maza, V; Esquivel Ojeda, J N; Gené Giralt, A; Luaces Cubells, C

2015-06-01

The aim of this study is to identify predictive factors of bacterial contamination in positive blood cultures (BC) collected in an emergency department. A prospective, observational and analytical study was conducted on febrile children aged on to 36 months, who had no risk factors of bacterial infection, and had a BC collected in the Emergency Department between November 2011 and October 2013 in which bacterial growth was detected. The potential BC contamination predicting factors analysed were: maximum temperature, time to positivity, initial Gram stain result, white blood cell count, absolute neutrophil count, band count, and C-reactive protein (CRP). Bacteria grew in 169 BC. Thirty (17.8%) were finally considered true positives and 139 (82.2%) false positives. All potential BC contamination predicting factors analysed, except maximum temperature, showed significant differences between true positives and false positives. CRP value, time to positivity, and initial Gram stain result are the best predictors of false positives in BC. The positive predictive values of a CRP value≤30mg/L, BC time to positivity≥16h, and initial Gram stain suggestive of a contaminant in predicting a FP, are 95.1, 96.9 and 97.5%, respectively. When all 3 conditions are applied, their positive predictive value is 100%. Four (8.3%) patients with a false positive BC and discharged to home were revaluated in the Emergency Department. The majority of BC obtained in the Emergency Department that showed positive were finally considered false positives. Initial Gram stain, time to positivity, and CRP results are valuable diagnostic tests in distinguishing between true positives and false positives in BC. The early detection of false positives will allow minimising their negative consequences. Copyright © 2014 Asociación Española de Pediatría. Published by Elsevier España, S.L.U. All rights reserved.

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.

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

Topical antibiotic treatment reduces tympanostomy tube biofilm formation.

PubMed

Thomas, Robert G; Ojano-Dirain, Carolyn; Antonelli, Patrick J

2011-05-01

Single doses of different ototopical antibiotic preparations (OAPs) have been shown to have an unequal reduction of post tympanostomy tube otorrhea (PTTO). Microbial biofilm formation on the tympanostomy tube (TT) has been implicated as one cause of PTTO. The goal of this study was to determine if TT exposure to a single dose of OAP reduces biofilm formation by Pseudomonas aeruginosa. Prospective and controlled. Fluoroplastic TTs were briefly exposed to plasma, followed by one of three OAPs (ofloxacin, neomycin/polymyxin B/hydrocortisone, or ciprofloxacin/dexamethasone) or saline (20 TT per group). TTs were placed in growth media with P. aeruginosa and incubated for 4 days, during which total bacterial growth was monitored by media turbidity. At 4 days, planktonic organisms were killed and biofilm development was measured with microbial counts. Bacterial growth was significantly delayed by OAPs, with the least growth seen with ciprofloxacin/dexamethasone followed by ofloxacin and neomycin/polymyxin B/hydrocortisone (P ≤ .0001). At day 4, bacterial growth was less with ciprofloxacin/dexamethasone than ofloxacin and neomycin/polymyxin B/hydrocortisone (P < .05). After 4 days, biofilm counts were lower on OAP-treated than saline-treated TTs (P = .0015) with both ciprofloxacin/dexamethasone and ofloxacin significantly less than saline (P < .05). Biofilm counts were not significantly different between OAPs (P > .05). Treatment of TTs with ototopical antibiotic preparations reduces P. aeruginosa growth and biofilm formation in vitro. This may, in part, explain the reduction of PTTO rates observed with single doses of OAPs. Copyright © 2011 The American Laryngological, Rhinological, and Otological Society, Inc.

N2 Gas Flushing Alleviates the Loss of Bacterial Diversity and Inhibits Psychrotrophic Pseudomonas during the Cold Storage of Bovine Raw Milk.

PubMed

Gschwendtner, Silvia; Alatossava, Tapani; Kublik, Susanne; Fuka, Mirna Mrkonjić; Schloter, Michael; Munsch-Alatossava, Patricia

2016-01-01

The quality and safety of raw milk still remains a worldwide challenge. Culture-dependent methods indicated that the continuous N2 gas-flushing of raw milk reduced the bacterial growth during cold storage by up to four orders of magnitude, compared to cold storage alone. This study investigated the influence of N2 gas-flushing on bacterial diversity in bovine raw-milk samples, that were either cold stored at 6°C or additionally flushed with pure N2 for up to one week. Next-generation sequencing (NGS) of the V1-V2 hypervariable regions of 16S rRNA genes, derived from amplified cDNA, which was obtained from RNA directly isolated from raw-milk samples, was performed. The reads, which were clustered into 2448 operational taxonomic units (OTUs), were phylogenetically classified. Our data revealed a drastic reduction in the diversity of OTUs in raw milk during cold storage at 6°C at 97% similarity level; but, the N2-flushing treatment alleviated this reduction and substantially limited the loss of bacterial diversity during the same cold-storage period. Compared to cold-stored milk, the initial raw-milk samples contained less Proteobacteria (mainly Pseudomonadaceae, Moraxellaceae and Enterobacteriaceae) but more Firmicutes (mainly Ruminococcaceaea, Lachnospiraceae and Oscillospiraceaea) and Bacteroidetes (mainly Bacteroidales). Significant differences between cold-stored and additionally N2-flushed milk were mainly related to higher levels of Pseudomononadaceae (including the genera Pseudomonas and Acinetobacter) in cold-stored milk samples; furthermore, rare taxa were better preserved by the N2 gas flushing compared to the cold storage alone. No major changes in bacterial composition with time were found regarding the distribution of the major 9 OTUs, that dominated the Pseudomonas genus in N2-flushed or non-flushed milk samples, other than an intriguing predominance of bacteria related to P. veronii. Overall, this study established that neither bacteria causing milk spoilage nor any well-known human pathogen or anaerobe benefited from the N2 gas flushing even though the N2-flushed and non-flushed cold-stored milk differed in bacterial counts by up to 104-fold.

N2 Gas Flushing Alleviates the Loss of Bacterial Diversity and Inhibits Psychrotrophic Pseudomonas during the Cold Storage of Bovine Raw Milk

PubMed Central

Kublik, Susanne; Fuka, Mirna Mrkonjić; Schloter, Michael; Munsch-Alatossava, Patricia

2016-01-01

The quality and safety of raw milk still remains a worldwide challenge. Culture-dependent methods indicated that the continuous N2 gas-flushing of raw milk reduced the bacterial growth during cold storage by up to four orders of magnitude, compared to cold storage alone. This study investigated the influence of N2 gas-flushing on bacterial diversity in bovine raw-milk samples, that were either cold stored at 6°C or additionally flushed with pure N2 for up to one week. Next-generation sequencing (NGS) of the V1-V2 hypervariable regions of 16S rRNA genes, derived from amplified cDNA, which was obtained from RNA directly isolated from raw-milk samples, was performed. The reads, which were clustered into 2448 operational taxonomic units (OTUs), were phylogenetically classified. Our data revealed a drastic reduction in the diversity of OTUs in raw milk during cold storage at 6°C at 97% similarity level; but, the N2-flushing treatment alleviated this reduction and substantially limited the loss of bacterial diversity during the same cold-storage period. Compared to cold-stored milk, the initial raw-milk samples contained less Proteobacteria (mainly Pseudomonadaceae, Moraxellaceae and Enterobacteriaceae) but more Firmicutes (mainly Ruminococcaceaea, Lachnospiraceae and Oscillospiraceaea) and Bacteroidetes (mainly Bacteroidales). Significant differences between cold-stored and additionally N2-flushed milk were mainly related to higher levels of Pseudomononadaceae (including the genera Pseudomonas and Acinetobacter) in cold-stored milk samples; furthermore, rare taxa were better preserved by the N2 gas flushing compared to the cold storage alone. No major changes in bacterial composition with time were found regarding the distribution of the major 9 OTUs, that dominated the Pseudomonas genus in N2-flushed or non-flushed milk samples, other than an intriguing predominance of bacteria related to P. veronii. Overall, this study established that neither bacteria causing milk spoilage nor any well-known human pathogen or anaerobe benefited from the N2 gas flushing even though the N2-flushed and non-flushed cold-stored milk differed in bacterial counts by up to 104-fold. PMID:26730711

Predictive value of decoy receptor 3 in postoperative nosocomial bacterial meningitis.

PubMed

Liu, Yong-Juan; Shao, Li-Hua; Wang, Qian; Zhang, Jian; Ma, Rui-Ping; Liu, Hai-Hong; Dong, Xiao-Meng; Ma, Li-Xian

2014-11-03

Nosocomial bacterial meningitis requires timely treatment, but what is difficult is the prompt and accurate diagnosis of this disease. The aim of this study was to assess the potential role of decoy receptor 3 (DcR3) levels in the differentiation of bacterial meningitis from non-bacterial meningitis. A total of 123 patients were recruited in this study, among them 80 patients being with bacterial meningitis and 43 patients with non-bacterial meningitis. Bacterial meningitis was confirmed by bacterial culture of cerebrospinal fluid (CSF) culture and enzyme-linked immunosorbent assay (ELISA) was used to detect the level of DcR3 in CSF. CSF levels of DcR3 were statistically significant between patients with bacterial meningitis and those with non-bacterial meningitis (p<0.001). A total of 48.75% of patients with bacterial meningitis received antibiotic>24 h before CSF sampling, which was much higher than that of non-bacterial meningitis. CSF leucocyte count yielded the highest diagnostic value, with an area under the receiver operating characteristic curve (ROC) of 0.928, followed by DcR3. At a critical value of 0.201 ng/mL for DcR3, the sensitivity and specificity were 78.75% and 81.40% respectively. DcR3 in CSF may be a valuable predictor for differentiating patients with bacterial meningitis from those with non-bacterial meningitis. Further studies are needed for the validation of this study.

Development of microbial spoilage and lipid and protein oxidation in rabbit meat.

PubMed

Nakyinsige, K; Sazili, A Q; Aghwan, Z A; Zulkifli, I; Goh, Y M; Abu Bakar, F; Sarah, S A

2015-10-01

This experiment aimed to determine microbial spoilage and lipid and protein oxidation during aerobic refrigerated (4°C) storage of rabbit meat. Forty male New Zealand white rabbits were slaughtered according to the Halal slaughter procedure. The hind limbs were used for microbial analysis while the Longissimus lumborum m. was used for determination of lipid and protein oxidation. Bacterial counts generally increased with aging time and the limit for fresh meat (10(8)cfu/g) was reached at d 7 postmortem. Significant differences in malondialdehyde content were observed after 3d of storage. The thiol concentration significantly decreased with increase in aging time. The band intensities of myosin heavy chain and troponin T significantly reduced with increased refrigerated storage while actin remained relatively stable. This study thus proposes protein oxidation as a potential deteriorative change in refrigerated rabbit meat along with microbial spoilage and lipid oxidation. Copyright © 2015 Elsevier Ltd. All rights reserved.

High-Throughput Particle Uptake Analysis by Imaging Flow Cytometry

PubMed Central

Smirnov, Asya; Solga, Michael D.; Lannigan, Joanne; Criss, Alison K.

2017-01-01

Quantifying the efficiency of particle uptake by host cells is important in fields including infectious diseases, autoimmunity, cancer, developmental biology, and drug delivery. Here we present a protocol for high-throughput analysis of particle uptake using imaging flow cytometry, using the bacterium Neisseria gonorrhoeae attached and internalized to neutrophils as an example. Cells are exposed to fluorescently labeled bacteria, fixed, and stained with a bacteria-specific antibody of a different fluorophore. Thus in the absence of a permeabilizing agent, extracellular bacteria are double-labeled with two fluorophores while intracellular bacteria remain single-labeled. A spot count algorithm is used to determine the number of single- and double-labeled bacteria in individual cells, to calculate the percent of cells associated with bacteria, percent of cells with internalized bacteria, and percent of cell-associated bacteria that are internalized. These analyses quantify bacterial association and internalization across thousands of cells and can be applied to diverse experimental systems. PMID:28369762

An evaluation of bacterial contamination of barriers used in periapical tissue regeneration: Part 2--Bacterial penetration.

PubMed

Sharma, Priya; Mickel, André K; Chogle, Sami; Sharma, Prem Nath; Han, Yiping W; Jones, Jefferson J

2008-03-01

To compare the relative penetration of Prevotella melaninogenica and Enterococcus faecalis through 3 guided tissue regeneration membranes: Atrisorb, Lambone, and OsseoQuest. It was hypothesized that OsseoQuest would show increased bacterial penetration when compared to Lambone and Atrisorb. Centrifuge tubes containing trypticase soy broth were sealed with circular sections of membranes and placed in test tubes containing culture media. The bacterial penetration was assessed by passage of bacteria from the outer tube culture media to the inner centrifuge tube media through the membrane. After incubation for 4 and 48 hours, the media from the outer and inner tubes were compared for bacterial count. P melaninogenica exhibited 91% penetration for Lambone in 2 days, while OsseoQuest displayed 87% penetration with E faecalis in the same time. Atrisorb displayed a minimal penetration with both bacteria (2%). Atrisorb displayed the least bacterial penetration, which may be attributed to membrane structure, chemical configuration, hydrophobicity, and porosity of tested membranes.

Erysipelas

MedlinePlus

... the next 24- 48 hours a red shiny plaque forms usually proximal to the area of entry into the skin. A dermatologist can usually make a diagnosis by examining the patient but sometimes other tests including bacterial skin culture, white blood cell count and blood ...

Culture-free, highly sensitive, quantitative detection of bacteria from minimally processed samples using fluorescence imaging by smartphone.

PubMed

Shrivastava, Sajal; Lee, Won-Il; Lee, Nae-Eung

2018-06-30

A critical unmet need in the diagnosis of bacterial infections, which remain a major cause of human morbidity and mortality, is the detection of scarce bacterial pathogens in a variety of samples in a rapid and quantitative manner. Herein, we demonstrate smartphone-based detection of Staphylococcus aureus in a culture-free, rapid, quantitative manner from minimally processed liquid samples using aptamer-functionalized fluorescent magnetic nanoparticles. The tagged S. aureus cells were magnetically captured in a detection cassette, and then fluorescence was imaged using a smartphone camera with a light-emitting diode as the excitation source. Our results showed quantitative detection capability with a minimum detectable concentration as low as 10 cfu/ml by counting individual bacteria cells, efficiently capturing S. aureus cells directly from a peanut milk sample within 10 min. When the selectivity of detection was investigated using samples spiked with other pathogenic bacteria, no significant non-specific detection occurred. Furthermore, strains of S. aureus from various origins showed comparable results, ensuring that the approach can be widely adopted. Therefore, the quantitative fluorescence imaging platform on a smartphone could allow on-site detection of bacteria, providing great potential assistance during major infectious disease outbreaks in remote and resource-limited settings. Copyright © 2018 Elsevier B.V. All rights reserved.

Inhibition of nitric oxide production and the effects of arginine and Lactobacillus administration in an acute liver injury model.

PubMed

Adawi, D; Molin, G; Jeppsson, B

1998-12-01

To study the effect of inhibiting nitric oxide production and the effects of arginine and lactobacilli administration in an acute liver injury (LI) model. Infectious complications caused by enteric bacteria are common in patients with liver diseases and those who have undergone liver surgery. Increased bacterial translocation has been proposed as one underlying mechanism. Lactobacilli constitute an integral part of the normal gastrointestinal microecology; they are involved in host metabolism and have many beneficial properties. Arginine has numerous roles in cellular metabolism and may be metabolized by lactobacilli in some cases. We have previously shown that rectal administration of Lactobacillus plantarum DSM 9843 (strain 299v), with and without arginine, in an acute LI model significantly reduces the extent of the LI and reduces bacterial translocation. To clarify the pathogenetic mechanisms, we studied the role of nitric oxide in the effects of L. plantarum and arginine in acute LI, as determined by bacterial translocation, ileal, cecal, and colonic nucleotides, RNA, and DNA. Male Sprague-Dawley rats were used. L. plantarum, 2% arginine, and/or N-nitro-L-arginine methyl ester (L-NAME), as appropriate, were administered rectally once daily for 8 days. Acute LI was induced on the eighth day by intraperitoneal injection of D-galactosamine (1.1 g/kg body weight), and samples were collected after 24 hours. Bacterial translocation was evaluated by culture of portal and arterial blood, mesenteric lymph nodes, and liver tissue. Liver enzymes and bilirubin were assayed in the serum. The bacterial load in the cecum and colon was determined. Ileal, cecal, and colonic mucosal nucleotides, RNA, and DNA were evaluated. The levels of liver enzymes and bilirubin were lower in liver-injured rats supplemented with arginine and Lactobacillus, and this effect was abolished by the addition of L-NAME. Inhibition of nitric oxide production (by L-NAME) increased bacterial translocation in many groups. L-NAME administration increased the cecal and colonic bacterial count and decreased the levels of mucosal nucleotides, RNA, and DNA. Inhibition of nitric oxide production modulated the effects of arginine and L. plantarum in this acute LI model. L-NAME potentiated the LI, as indicated by elevation of liver enzymes and bilirubin, and it also increased bacterial translocation and the cecal and colonic bacterial count. Increased bacterial translocation could be one of the mechanisms by which LI is potentiated.

Influence of storage and preservation on microbiological quality of silo ovine milk.

PubMed

de Garnica, M L; Santos, J A; Gonzalo, C

2011-04-01

This study was designed to analyze the effects of the storage and preservation conditions on counts of mesophilic, thermoduric, psychotrophic, coliform, Escherichia coli, Streptococcus agalactiae, and Staphylococcus aureus organisms in silo ovine milk. A total of 910 analytical determinations were conducted from aliquots of 10 silo ovine milks. The conditions tested were unpreserved and azidiol-preserved milk stored at 4°C, and unpreserved milk stored at -20°C. Milk aged 2, 24, 48, 72, and 96 h post-collection for refrigerated aliquots, and 7, 15, and 30 d post-collection for frozen aliquots. The factors silo and storage conditions significantly contributed to variation of all microbiological variables, although milk age effect within storage was only significant for mesophilic, psychrotrophic, and coliform bacteria counts. In refrigerated raw milk, mesophile, psychrotroph, and coliform counts significantly increased over 96 h post-collection, whereas the other groups and bacteria species tested maintained their initial concentration. In all cases, azidiol preservation maintained the initial bacterial concentration in raw sheep milk under refrigeration throughout 96 h. Thus, azidiol was a suitable preservative for microbiological studies in sheep milk. Smallest counts were registered for frozen samples, particularly for coliforms, E. coli, Strep. agalactiae and Staph. aureus. Estimates of mesophilic, thermoduric and psychrotrophic organisms showed similar values on both azidiol-preserved and frozen milk samples. Coliforms and E. coli counts significantly decrease over time after freezing. Consequently, freezing at -20°C could also be appropriate for analysis of mesophilic, thermoduric, and psychrotrophic bacterial groups, but not for coliforms or mammary pathogens. Copyright © 2011 American Dairy Science Association. Published by Elsevier Inc. All rights reserved.

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.

Bacteriological Quality of Foods and Water Sold by Vendors and in Restaurants in Nsukka, Enugu State, Nigeria: A Comparative Study of Three Microbiological Methods

PubMed Central

Ibe, Nnenne I.; Iroegbu, Christian U.

2011-01-01

Bacterial count in prepared food or water is a key factor in assessing the quality and safety of food. It also reveals the level of hygiene adopted by food handlers in the course of preparation of such foods. This comparative study evaluated the bacteriological quality of food and water consumed in Nsukka, Enugu state, Nigeria, using three bacteria enumeration methods. Data obtained are assumed to reflect the level of personal and environmental hygiene in the study population. Ten types of foods—beans, yam, abacha, okpa, moimoi, pear, cassava foofoo, rice, agidi, and garri—and 10 water samples were evaluated for bacteriological quality, precisely determining the level of coliform contamination, using the most probable number (MPN), lactose fermentation count (LFC), and Escherichia coli count (ECC) methods. Bacterial counts differed significantly (p<0.05) among the various food samples. However, this did not differ significantly in the three methods used for the enumeration of coliforms, suggesting that any of the three methods could be validly used for such studies with confidence. Escherichia coli and Klebsiella pneumoniae were the two major coliforms identified among 98 coliform isolates obtained from the various food samples, of which 78 (79.6%) were assumed to be of human origin on account of their ability to grow at 44 °C. The level of coliform contamination in the food samples from vendors and restaurants (geometric mean count 7.64-9.21; MPN ≥50) were above the accepted 104 colony-forming unit/g or MPN ≤10 limits. The results of the study, therefore, call for stringent supervision and implementation of food-safety practices and regular education on food and personal hygiene among food vendors. PMID:22283029

Antimicrobial effects of pepper, parsley, and dill and their roles in the microbiological quality enhancement of traditional Egyptian Kareish cheese.

PubMed

Wahba, Nahed M; Ahmed, Amany S; Ebraheim, Zedan Z

2010-04-01

This study was designed to assess the application of some edible plants including cayenne, green pepper, parsley, and dill to Kareish cheese and to evaluate the antimicrobial activity of these plant materials against natural microflora, coliforms, molds, and Staphylococcus aureus. Twelve different concentrations of ethanol extract of the plants were prepared for determination of the minimal inhibitory concentration. Cayenne and green pepper extracts showed highest activity followed by dill and parsley against S. aureus. Addition of cayenne or green pepper to Kareish cheese during manufacture revealed that both plants were able reduce the S. aureus population to undetectable level within the first and second days of storage. To study the effect of combining plant materials on the microbiological quality of ready-to-eat Kareish cheese, the total bacterial count, coliform count, and yeast and molds counts were determined. It has been found that addition of plant materials to Kareish cheese reduced the total bacterial and coliform populations. All concentrations of cayenne, green pepper, dill, and parsley (9%) completely reduced the yeast count within 2 hours. Cayenne and green pepper completely reduced the mold count within 2 days, whereas parsley and dill were found to be less effective. Kareish cheese prepared with 1% cayenne pepper and 3% and 6% each of green pepper, dill, and parsley were found strongly acceptable to the consumer and considered the most preferable type. Therefore, this study revealed that pepper, parsley, and dill exhibited antibacterial activity against natural microflora, coliforms, yeast and molds, and S. aureus in Kareish cheese, and the addition of these plants is acceptable to the consumer and may contribute to the development of new and safe varieties of Kareish cheese.

Relationships Among Microbial Communities, Maternal Cells, Oligosaccharides, and Macronutrients in Human Milk.

PubMed

Williams, Janet E; Price, William J; Shafii, Bahman; Yahvah, Katherine M; Bode, Lars; McGuire, Mark A; McGuire, Michelle K

2017-08-01

Human milk provides all essential nutrients necessary for early life and is rich in nonnutrients, maternally derived (host) cells, and bacteria, but almost nothing is known about the interplay among these components. Research aim: The primary objective of this research was to characterize relationships among macronutrients, maternal cells, and bacteria in milk. Milk samples were collected from 16 women and analyzed for protein, lipid, fatty acid, lactose, and human milk oligosaccharide concentrations. Concentrations of maternal cells were determined using microscopy, and somatic cell counts were enumerated. Microbial ecologies were characterized using culture-independent methods. Absolute and relative concentrations of maternal cells were mostly consistent within each woman as were relative abundances of bacterial genera, and there were many apparent relationships between these factors. For instance, relative abundance of Serratia was negatively associated with somatic cell counts ( r = -.47, p < .0001) and neutrophil concentration ( r = -.38, p < .0006). Concentrations of several oligosaccharides were correlated with maternally derived cell types as well as somatic cell counts; for example, lacto-N-tetraose and lacto-N-neotetraose were inversely correlated with somatic cell counts ( r = -.64, p = .0082; r = -.52, p = .0387, respectively), and relative abundance of Staphylococcus was positively associated with total oligosaccharide concentration ( r = .69, p = .0034). Complex relationships between milk nutrients and bacterial community profile, maternal cells, and milk oligosaccharides were also apparent. These data support the possibility that profiles of maternally derived cells, nutrient concentrations, and the microbiome of human milk might be interrelated.

Microbiological evaluation of groups of beef carcasses: heifers and steers.

PubMed Central

Jericho, K W; Bradley, J A; Kozub, G C

1994-01-01

Numbers of mesophilic bacteria were estimated on carcasses of 25 heifers and 25 steers of beef breeds in a modern, high-line-speed abattoir. One side of each carcass from each sex was sampled at the end of the kill-floor, before the carcass wash, on each of 25 visits. Two adjacent excision samples (5 x 5 x 0.5 cm) were taken from each of ten sites and processed for automatic enumeration of aerobic bacteria on hydrophobic grid membrane filters. The effects of sex and carcass weight on bacterial counts were examined. Groups of carcasses were examined to determine the sample size required for future assessments of kill-floor hygiene. The log10 of the most probable number of growth units (MPNGU)/cm2 did not differ significantly between heifers and steers (average over the ten sites of 2.2) and there was no effect of carcass weight on bacterial counts for nine of the ten sites. There were, however, highly significant (p < 0.001) differences in the counts between sites and the counts from the ten sites clustered into five homogenous groups. The between-carcass component of variation at a site was generally larger than the within-carcass component. We conclude that, to estimate the mean log10 MPNGU/cm2 at a site to within +/- 0.5 units, future group-carcass evaluations require about 200 samples from 10 (two adjacent samples/site) or 20 carcasses (one sample/site). PMID:7954120

Neutrophil-to-lymphocyte ratio in the differential diagnosis of acute bacterial meningitis.

PubMed

Mentis, A-F A; Kyprianou, M A; Xirogianni, A; Kesanopoulos, K; Tzanakaki, G

2016-03-01

The differential diagnosis of acute community-acquired meningitis is of paramount importance in both therapeutic and healthcare-related economic terms. Despite the routinely used markers, novel, easily calculated, and rapidly available biomarkers are needed particularly in resource-poor settings. A promising, exponentially studied inflammatory marker is the neutrophil-to-lymphocyte ratio (NLR), albeit not assessed in meningitis. The aim of this study was to investigate the utility of the NLR in the differential diagnosis of acute meningitis. Data on cerebrospinal fluid (CSF) and blood leukocyte parameters from more than 4,000 patients diagnosed with either bacterial or viral meningitis in Greece during the period 2006-2013 were retrospectively examined. The diagnostic accuracy of the NLR and neutrophil counts in CSF and blood were evaluated by receiver operating characteristic curves. The discrimination ability of both the NLR and neutrophil counts was significantly higher in CSF than in blood. The optimal cutoff values of the NLR and neutrophil counts were 2 in CSF vs 8 in blood, and 287 cells in CSF vs 12,100 cells in blood, respectively. For these values, sensitivity, negative predictive value, and odds ratio were statistically significantly higher in CSF than blood for both markers. Logistic regression analysis showed that the CSF NLR carries independent and additive information to neutrophil counts in the differential diagnosis of acute meningitis. This study is the first one to assess NLR in acute meningitis, providing promising results for its differential diagnosis.

Photodynamic therapy on bacterial reduction in dental caries: in vivo study

NASA Astrophysics Data System (ADS)

Baptista, Alessandra; Araujo Prates, Renato; Kato, Ilka Tiemy; Amaral, Marcello Magri; Zanardi de Freitas, Anderson; Simões Ribeiro, Martha

2010-04-01

The reduction of pathogenic microorganisms in supragingival plaque is one of the principal factors in caries prevention and control. A large number of microorganisms have been reported to be inactivated in vitro by photodynamic therapy (PDT). The purpose of this study was to develop a rat model to investigate the effects of PDT on bacterial reduction in induced dental caries. Twenty four rats were orally inoculated with Streptococcus mutans cells (ATCC 25175) for three consecutive days. The animals were fed with a cariogenic diet and water with 10% of sucrose ad libitum, during all experimental period. Caries lesion formation was confirmed by Optical Coherence Tomography (OCT) 5 days after the beginning of the experiment. Then, the animals were randomly divided into two groups: Control Group: twelve animals were untreated by either light or photosensitizer; and PDT Group: twelve animals were treated with 100μM of methylene blue for 5min and irradiated by a Light Emitting Diode (LED) at λ = 640+/-30nm, fluence of 172J/cm2, output power of 240mW, and exposure time of 3min. Microbiological samples were collected before, immediately after, 3, 7 and 10 days after treatment and the number of total microaerophiles was counted. OCT images showed areas of enamel demineralization on rat molars. Microbiological analysis showed a significant bacterial reduction after PDT. Furthermore, the number of total microaerophiles in PDT group remained lower than control group until 10 days posttreatment. These findings suggest that PDT could be an alternative approach to reduce bacteria in dental caries.

Microbial adherence to cosmetic contact lenses.

PubMed

Chan, Ka Yin; Cho, Pauline; Boost, Maureen

2014-08-01

To investigate whether cosmetic contact lenses (CCL) with surface pigments affect microbial adherence. Fifteen brands of CCL were purchased from optical, non-optical retail outlets, and via the Internet. A standardized rub-off test was performed on each CCL (five lenses per brand) to confirm the location of the pigments. The rub-off test comprised gentle rubbing on the surfaces of each CCL with wetted cotton buds for a maximum of 20 rubs per surface. A new set of CCL (five lenses per brand) were incubated in Pseudomonas aeruginosa overnight. Viable counts of adhered bacteria were determined by the number of colony-forming units (CFU) on agar media on each lens. The adherence of P. aeruginosa as well as Staphylococcus aureus and Serratia marcescens to three brands of CCL (A-C) (five lenses per brand) were also compared to their adherences on their clear counterparts. Only two of the 15 brands of CCL tested (brands B and C) had pigments that did not detach with the rub-off test. The remaining 13 brands of CCL all failed the rub-off test and these lenses showed higher P. aeruginosa adherence (8.7 × 10(5)-1.9 × 10(6) CFU/lens). Brands B and C lenses showed at least six times less bacterial adhesion than the other 13 brands. Compared to their clear counterparts, bacterial adherence to brands B and C lenses did not differ significantly, whereas brand A lenses showed significantly higher adherence. Surface pigments on CCL resulted in significantly higher bacterial adherence. Copyright © 2013 British Contact Lens Association. Published by Elsevier Ltd. All rights reserved.

Bacteriuria and antibiotic resistance in catheter urine specimens following radical prostatectomy.

PubMed

Banks, Jessica A; McGuire, Barry B; Loeb, Stacy; Shrestha, Sanjina; Helfand, Brian T; Catalona, William J

2013-10-01

There are increasing reports of infectious complications following prostate biopsy due to fluoroquinolone resistance. To determine infectious complications at catheter removal following radical prostatectomy (RP), another setting in daily urological practice where fluoroquinolone prophylaxis is frequently used. We prospectively examined urine culture results collected from 334 RP patients immediately prior to catheter removal. Patients received prophylactic antibiotics 1 day before, the day of, and for 5 days after catheter removal. Culture results were reviewed for bacterial species and antimicrobial susceptibilities. Patients with positive urine cultures resistant to the prophylactic antibiotic were switched to culture-specific antibiotic therapy and underwent follow-up culture. The frequency of urinary tract infection (UTI), complications, additional antibiotic therapy, and repeat urine cultures was determined within 60 days. Of the 334 patients identified, 203 (61%) had cultures with no bacterial growth, and 48 (14%) had colony counts of <1,000 bacteria or Candida albicans and received no further antibiotics. The remaining 83 (25%) had positive culture results, of which 7% were resistant to ciprofloxacin. Twenty-four bacterial species were identified, with Pseudomonas aeruginosa (5%) Escherichia coli (4%), and Staphylococcus epidermidis (3%) being the most frequent. Only two (0.6%) men developed clinical symptoms consistent with UTI (i.e., suprapubic pain, fever) prior to catheter removal, and no serious complications occurred. A substantial proportion of RP patients have positive urine cultures at the time of catheter removal, despite the administration of prophylactic fluoroquinolone antibiotics. Potentially virulent organisms are commonly cultured, and ciprofloxacin resistance is frequent. However, outcomes are favorable when culture-specific oral antibiotic therapy is initiated. Copyright © 2013 Elsevier Inc. All rights reserved.

Sperm Quality during Storage Is Not Affected by the Presence of Antibiotics in EquiPlus Semen Extender but Is Improved by Single Layer Centrifugation

PubMed Central

Spergser, Joachim; Kuhl, Juliane; Schmidt, Kathrin; Johannisson, Anders

2017-01-01

Contamination of semen with bacteria arises during semen collection and handling. This bacterial contamination is typically controlled by adding antibiotics to semen extenders but intensive usage of antibiotics can lead to the development of bacterial resistance and may be detrimental to sperm quality. The objective of this study was to determine the effects of antibiotics in a semen extender on sperm quality and to investigate the effects of removal of bacteria by modified Single Layer Centrifugation (MSLC) through a colloid. Semen was collected from six adult pony stallions (three ejaculates per male). Aliquots of extended semen were used for MSLC with Equicoll, resulting in four treatment groups: control and MSLC in extender with antibiotics (CA and SA, respectively); control and MSLC in extender without antibiotics (CW and SW, respectively). Sperm motility, membrane integrity, mitochondrial membrane potential and chromatin integrity were evaluated daily by computer-assisted sperm analysis (CASA) and flow cytometry. There were no differences in sperm quality between CA and CW, or between SA and SW, although progressive motility was negatively correlated to total bacterial counts at 0 h. However, MSLC groups showed higher mean total motility (P < 0.001), progressive motility (P < 0.05), membrane integrity (P < 0.0001) and mitochondrial membrane potential (P < 0.05), as well as better chromatin integrity (P < 0.05), than controls. Sperm quality remained higher in the MSLC groups than controls throughout storage. These results indicate that sperm quality was not adversely affected by the presence of antibiotics but was improved considerably by MSLC. PMID:29267226

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.

Influence of Biopreparations on the Bacterial Community of Oily Waste

NASA Astrophysics Data System (ADS)

Biktasheva, L. R.; Galitskaya, P. Yu; Selivanovskaya, S. Yu

2018-01-01

Oil pollution is reported to be one the most serious environmental problems nowadays. Therefore, methods of remediation of oily polluted soils and oily wastes are of great importance. Bioremediation being a perspective method of sanitation of oil pollutions, includes biostimulation of the polluted sites’ indigenous microflora, and in some cases additional introduction of active strains able to decompose hydrocarbon. The efficacy of introducing such biopreparations depends on the interactions between the introduced microbes and the indigenous ones. In this study, the influence of bacterial consortium (Rhodococcus jialingiae, Stenotrophomonas rhizophila and Pseudomonas gessardii) introduction on the bioremediation of an oily waste sampled from a refinery situated in the Mari El region (Russia) was estimated. Single and multiple inoculations of the consortium in addition to moistening and aeration were compared with a control sample, which included only aeration and moistening of the waste. It was shown, that two of the three introduced strains (Rh. jialingiae and Ps.gessardii) gene copy numbers were higher in the inoculated variants than in the control sample and with their initial counts, which meant that these strains survived and included into the bacterial community of the wastes. At the same time, bacterial counts were significantly lower, and the physiological profile of waste microflora slightly altered in the inoculated remediation variants as compared with the control sample. Interestingly, no difference in the degradation rates of hydrocarbons was revealed in the inoculated remediation variants and the control sample.

New Bacterial Infection in the Prostate after Transrectal Prostate Biopsy.

PubMed

Seo, Yumi; Lee, Gilho

2018-04-23

The prostate is prone to infections. Hypothetically, bacteria can be inoculated into the prostate during a transrectal prostate biopsy (TRPB) and progress into chronic bacterial prostatitis. Therefore, we examined new bacterial infections in biopsied prostates after TRPB and whether they affect clinical characteristics in the biopsied patients. Of men whose prostate cultures have been taken prior to TRPB, 105 men with bacteria-free benign prostate pathology underwent an additional repeated prostate culture within a year after TRPB. Twenty out of 105 men (19.05%) acquired new bacteria in their naïve prostates after TRPB. Except for one single case of Escherichia coli infection, 19 men had acquired gram-positive bacteria species. Between the culture-positive and negative groups, there were no significant differences in age, serum prostate-specific antigen (PSA) level, white blood cell (WBC) counts in expressed prostatic secretion (EPS), prostate volume, symptom severities in Korean version of the National Institutes of Health-Chronic Prostatitis Symptom Index (NIH-CPSI) questionnaire, and patient-specific risk factors for biopsy associated infectious complications. Additionally, the TRPB procedure increased the WBC counts in post-biopsy EPS ( P = 0.031, McNemar test), but did not increase the serum PSA level and symptoms of NIH-CPSI in 20 men who acquired new bacteria after TRPB. The TRPB procedure was significantly associated with acquiring new bacterial infections in the biopsied prostate, but these localized bacteria did not affect patients' serum PSA level and symptoms after biopsy.

Microbiological quality of cooked foods and drinks sold in higher educational institutions around Yala, Pattani, and Narathiwat Provinces, Southern Thailand

NASA Astrophysics Data System (ADS)

Dalee, Abdullah D.; Sali, Khosiya; Hayeeyusoh, Nurainee; Hayeewangoh, Zubaidah; Thadah, Amporn

2017-08-01

Quality of cooked foods and drinks water sold within the vicinity of higher institutions located in Yala, Pattani and Narathiwat provinces were randomly sampled and microbiologically evaluated. As to Thai National Food Safety Standard, various food menu and drinks were subjected to conventionally determining the bacterial index; Most Probable Number (MPN) of coliform and fecal coliform as well as the detection of indicator organisms; Escherichia coli, Staphylococcus aureus and Salmonella sp. As for bacterial index, results showed that curry-type likes PSU-stir-fried liver (7.5x106 CFU/g), and and the lowest was PSU-Koleh chicken Roast (1.72x103 CFU/g). The highest and lowest counts of soup-type items were observed in YPH-KaengSom soup (1.9x107 CFU/g), and PSU-Palo soup (0.4x103 CFU/g), respectively. Higher bacterial counts were also found in YPH-spicy stir-fried chicken (7.5 x 106 CFU/g), and YPH-squid salad (2.2x107 CFU/g). For drinks, bacterial count ranged 2.0 x 103 to 8.3 x 103 CFU/g, and NRU-iced grape juice having the highest bacterial count (2.0x106 CFU/g). Overall, foods not complying to the Thai National Food Safety Standard of 1 x 103 CFU/g from higher to lower were those of soup, stir-fried, salad, fried, and curry categories with as much as 4:17 (23.53%), 4:21 (19.05%), 2:11 (18.18%), 2:16 (12.5%) and 1:12 (8.33%), respectively. As for Coliform and fecal coliform, the highest (>1100 MPN/g) and the lowest (0.34 MPN/g),were not much found in all food categories with percentages of 23.53, 24.00, 13.79, 9.10, and 47.37 for curry (4:17), soup (6:15), stir-fried (4:29), fried (2:22), and salad (9:19), respectively. However, indicator organisms were not detected in almost all food samples except PSU-chicken yellowish curry, NRU-chicken TongYam soup, NRU-Long-tail tuna soup, NRU-KaengSom soup, YPE-watery soup, NRU-stir-fried liver, NRU-omelets, NRU-fried chicken, YPE-crispy fish salad, and NRU-salted eggs salad, which showed the presence of E. coli, but not Staphylococcus aureus and Salmonella sp. As for drinks, 3 out of 40 samples (7.5%) were E. coli detectable, and they were YPE-water, YPE-iced tea, and YPH-iced tea. Evaluating results of such items of foods and drinks deem academically valuable, and may effectively raise public awareness and national monitoring in future.

Is it Possible to Sanitize Athletes' Shoes?

PubMed Central

Messina, Gabriele; Burgassi, Sandra; Russo, Carmela; Ceriale, Emma; Quercioli, Cecilia; Meniconi, Cosetta

2015-01-01

Context: Footwear should be designed to avoid trauma and injury to the skin of the feet that can favor bacterial and fungal infections. Procedures and substances for sanitizing the interior of shoes are uncommon but are important aspects of primary prevention against foot infections and unpleasant odor. Objective: To evaluate the efficacy of a sanitizing technique for reducing bacterial and fungal contamination of footwear. Design: Crossover study. Setting: Mens Sana basketball team. Patients or Other Participants: Twenty-seven male athletes and 4 coaches (62 shoes). Intervention(s): The experimental protocol required a first sample (swab), 1/shoe, at time 0 from inside the shoes of all athletes before the sanitizing technique began and a second sample at time 1, after about 4 weeks, April 2012 to May 2012, of daily use of the sanitizing technique. Main Outcome Measure(s): The differences before and after use of the sanitizing technique for total bacterial count at 36°C and 22°C for Staphylococcus spp, yeasts, molds, Enterococcus spp, Pseudomonas spp, Escherichia coli, and total coliform bacteria were evaluated. Results: Before use of the sanitizing technique, the total bacterial counts at 36°C and 22°C and for Staphylococcus spp were greater by a factor of 5.8 (95% confidence interval [CI] = 3.42, 9.84), 5.84 (95% CI = 3.45, 9.78), and 4.78 (95% CI = 2.84, 8.03), respectively. All the other comparisons showed a reduction in microbial loads, whereas E coli and coliforms were no longer detected. No statistically significant decrease in yeasts (P = .0841) or molds (P = .6913) was recorded probably because of low contamination. Conclusions: The sanitizing technique significantly reduced the bacterial presence in athletes' shoes. PMID:25415415

The influence of ultrasonic treatment on the growth of the strains of Salmonella enterica subs. typhimurium.

PubMed

Sienkiewicz, Jolanta Joanna; Wesołowski, Andrzej; Stankiewicz, Wanda; Kotowski, Romuald

2017-07-01

This study proposes the destruction of pathogenic bacteria with the use of ultrasound waves because the more commonly used thermal processing methods often result in lowering the nutritional value of food. The study presents the impact of ultrasound of 20, 40 and 100 kHz frequencies and the power of 10.5 W/cm 2 on the growth of the strain of Salmonella enterica subs. typhimurium . The tests were carried out both in chilled and non-chilled treatment mediums, with an average bacterial population >10 5 and >10 8  CFU/cm 3 . The total inactivation of Salmonella spp. was observed in the tests in the low-population non-chilled treatment medium after sonication at 20 and 40 kHz for 30 min, and in high bacterial population at 20 kHz for 30 min. A reduction in the average number of bacteria was reported in the low-population non-chilled medium after 15 min of sonication at 20, 40 and 100 kHz; after 15 min of sonication at 20 and 100 kHz of the material of high bacterial population; and in the low-population chilled treatment mediums after 15 and 30 min at 20 kHz. The samples with inactivated bacteria and those with reduced bacterial counts maintained the same levels when stored at 4 °C for 24 and 48 h. Bacteria inactivation obtained after sonication lasted for up to 48 h in storage at 21 °C. For the samples with reduced bacterial counts stored at 21 °C, a rise in the average number of bacteria was recorded.

Evaluating the effect of intraoperative peritoneal lavage on bacterial culture in dogs with suspected septic peritonitis.

PubMed

Swayne, Seanna L; Brisson, Brigitte; Weese, J Scott; Sears, William

2012-09-01

This pilot study describes the effect of intraoperative peritoneal lavage (IOPL) on bacterial counts and outcome in clinical cases of septic peritonitis. Intraoperative samples were cultured before and after IOPL. Thirty-three dogs with presumed septic peritonitis on the basis of cytology were managed surgically during the study period. Positive pre-lavage bacterial cultures were found in 14 cases, 13 of which were a result of intestinal leakage. The post-lavage cultures showed fewer isolates in 9 cases and in 1 case became negative. The number of dogs with a decrease in the concentration of bacteria cultured from pre-lavage to post-lavage samples was not statistically significant. There was no significant effect of the change in pre- to post-lavage culture, single versus multiple types of bacteria, selection of an appropriate empiric antimicrobial on survival or the need for subsequent surgery. This pilot study describes the effect of intraoperative peritoneal lavage (IOPL) on bacterial counts and outcome in clinical cases of septic peritonitis. Intraoperative samples were cultured before and after IOPL. Thirty-three dogs with presumed septic peritonitis on the basis of cytology were managed surgically during the study period. Positive pre-lavage bacterial cultures were found in 14 cases, 13 of which were a result of intestinal leakage. The post-lavage cultures showed fewer isolates in 9 cases and in 1 case became negative. The number of dogs with a decrease in the concentration of bacteria cultured from pre-lavage to post-lavage samples was not statistically significant. There was no significant effect of the change in pre- to post-lavage culture, single versus multiple types of bacteria, selection of an appropriate empiric antimicrobial on survival or the need for subsequent surgery.

Cefepime and amikacin synergy in vitro and in vivo against a ceftazidime-resistant strain of Enterobacter cloacae.

PubMed

Mimoz, O; Jacolot, A; Padoin, C; Tod, M; Samii, K; Petitjean, O

1998-03-01

The activities of cefepime and amikacin alone or in combination against an isogenic pair of Enterobacter cloacae strains (wild type and stably derepressed, ceftazidime-resistant mutant) were compared using an experimental model of pneumonia in non-leucopenic rats. Animals were infected by administering 8.4 log10 cfu of E. cloacae intratracheally, and therapy was initiated 12 h later. At that time, the animals' lungs showed bilateral pneumonia and contained more than 7 log10 E. cloacae cfu/g tissue. Because rats eliminate amikacin and cefepime much more rapidly than humans, renal impairment was induced in all animals to simulate the pharmacokinetic parameters of humans. In-vitro susceptibilities showed an inoculum effect with cefepime proportional to the bacterial titre against the two strains, but more pronounced with the stably derepressed mutant strain, whereas with bacterial concentrations of up to 7 log10 cfu/mL, no inoculum effect was observed with amikacin. In-vitro killing indicated that antibiotic combinations were synergic only at intermediate concentrations. At peak concentrations, the combination was merely as effective as amikacin alone. At trough concentrations, a non-significant trend towards the superiority of the combination over each antibiotic alone was noted. Moreover, cefepime was either bacteriostatic or permitted regrowth of the organisms in the range of antibiotic concentrations tested. Although each antibiotic alone failed to decrease bacterial counts in the lungs, regardless of the susceptibility of the strain used, the combination of both antibiotics was synergic and induced a significant decrease in the lung bacterial count 24 h after starting therapy when compared with tissue bacterial numbers in untreated animals or animals treated with either antibiotic alone. No resistant clones emerged during treatment with any of the antibiotic regimens studied.

Predictive Value of Decoy Receptor 3 in Postoperative Nosocomial Bacterial Meningitis

PubMed Central

Liu, Yong-Juan; Shao, Li-Hua; Wang, Qian; Zhang, Jian; Ma, Rui-Ping; Liu, Hai-Hong; Dong, Xiao-Meng; Ma, Li-Xian

2014-01-01

Nosocomial bacterial meningitis requires timely treatment, but what is difficult is the prompt and accurate diagnosis of this disease. The aim of this study was to assess the potential role of decoy receptor 3 (DcR3) levels in the differentiation of bacterial meningitis from non-bacterial meningitis. A total of 123 patients were recruited in this study, among them 80 patients being with bacterial meningitis and 43 patients with non-bacterial meningitis. Bacterial meningitis was confirmed by bacterial culture of cerebrospinal fluid (CSF) culture and enzyme-linked immunosorbent assay (ELISA) was used to detect the level of DcR3 in CSF. CSF levels of DcR3 were statistically significant between patients with bacterial meningitis and those with non-bacterial meningitis (p < 0.001). A total of 48.75% of patients with bacterial meningitis received antibiotic >24 h before CSF sampling, which was much higher than that of non-bacterial meningitis. CSF leucocyte count yielded the highest diagnostic value, with an area under the receiver operating characteristic curve (ROC) of 0.928, followed by DcR3. At a critical value of 0.201 ng/mL for DcR3, the sensitivity and specificity were 78.75% and 81.40% respectively. DcR3 in CSF may be a valuable predictor for differentiating patients with bacterial meningitis from those with non-bacterial meningitis. Further studies are needed for the validation of this study. PMID:25372942

Genome-Wide Transcriptional Profiling of Clostridium perfringens SM101 during Sporulation Extends the Core of Putative Sporulation Genes and Genes Determining Spore Properties and Germination Characteristics.

PubMed

Xiao, Yinghua; van Hijum, Sacha A F T; Abee, Tjakko; Wells-Bennik, Marjon H J

2015-01-01

The formation of bacterial spores is a highly regulated process and the ultimate properties of the spores are determined during sporulation and subsequent maturation. A wide variety of genes that are expressed during sporulation determine spore properties such as resistance to heat and other adverse environmental conditions, dormancy and germination responses. In this study we characterized the sporulation phases of C. perfringens enterotoxic strain SM101 based on morphological characteristics, biomass accumulation (OD600), the total viable counts of cells plus spores, the viable count of heat resistant spores alone, the pH of the supernatant, enterotoxin production and dipicolinic acid accumulation. Subsequently, whole-genome expression profiling during key phases of the sporulation process was performed using DNA microarrays, and genes were clustered based on their time-course expression profiles during sporulation. The majority of previously characterized C. perfringens germination genes showed upregulated expression profiles in time during sporulation and belonged to two main clusters of genes. These clusters with up-regulated genes contained a large number of C. perfringens genes which are homologs of Bacillus genes with roles in sporulation and germination; this study therefore suggests that those homologs are functional in C. perfringens. A comprehensive homology search revealed that approximately half of the upregulated genes in the two clusters are conserved within a broad range of sporeforming Firmicutes. Another 30% of upregulated genes in the two clusters were found only in Clostridium species, while the remaining 20% appeared to be specific for C. perfringens. These newly identified genes may add to the repertoire of genes with roles in sporulation and determining spore properties including germination behavior. Their exact roles remain to be elucidated in future studies.

Genome-Wide Transcriptional Profiling of Clostridium perfringens SM101 during Sporulation Extends the Core of Putative Sporulation Genes and Genes Determining Spore Properties and Germination Characteristics

PubMed Central

Xiao, Yinghua; van Hijum, Sacha A. F. T.; Abee, Tjakko; Wells-Bennik, Marjon H. J.

2015-01-01

The formation of bacterial spores is a highly regulated process and the ultimate properties of the spores are determined during sporulation and subsequent maturation. A wide variety of genes that are expressed during sporulation determine spore properties such as resistance to heat and other adverse environmental conditions, dormancy and germination responses. In this study we characterized the sporulation phases of C. perfringens enterotoxic strain SM101 based on morphological characteristics, biomass accumulation (OD600), the total viable counts of cells plus spores, the viable count of heat resistant spores alone, the pH of the supernatant, enterotoxin production and dipicolinic acid accumulation. Subsequently, whole-genome expression profiling during key phases of the sporulation process was performed using DNA microarrays, and genes were clustered based on their time-course expression profiles during sporulation. The majority of previously characterized C. perfringens germination genes showed upregulated expression profiles in time during sporulation and belonged to two main clusters of genes. These clusters with up-regulated genes contained a large number of C. perfringens genes which are homologs of Bacillus genes with roles in sporulation and germination; this study therefore suggests that those homologs are functional in C. perfringens. A comprehensive homology search revealed that approximately half of the upregulated genes in the two clusters are conserved within a broad range of sporeforming Firmicutes. Another 30% of upregulated genes in the two clusters were found only in Clostridium species, while the remaining 20% appeared to be specific for C. perfringens. These newly identified genes may add to the repertoire of genes with roles in sporulation and determining spore properties including germination behavior. Their exact roles remain to be elucidated in future studies. PMID:25978838

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...

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...

Mutacins and bacteriocins like genes in Streptococcus mutans isolated from participants with high, moderate, and low salivary count.

PubMed

Soto, Carolina; Padilla, Carlos; Lobos, Olga

2017-02-01

To detect S. mutans producers of mutacins and bacteriocins like substances (BLIS) from saliva of participants with low, moderate, and high salivary counts. 123 strains of S. mutans were obtained from participants with low, moderate, and high salivary counts (age 18 and 20 years old) and their antibacterial capacity analyzed. By using PCR amplification, the expression levels of mutacins and BLIS genes were studied (expressed in arbitrary units/ml) in all three levels. S. mutans strains from participants with low salivary counts show high production of mutacins (63%). In contrast, participants with moderate and high salivary counts depict relatively low levels of mutacins (22 and 15%, respectively). Moreover, participants with low salivary counts showed high expression levels of genes encoding mutacins, a result that correlates with the strong antimicrobial activity of the group. Participants with moderate and high salivary counts however depict low expression levels of mutacin related genes, and little antimicrobial activity. No BLIS were detected in any of the groups studied. S. mutans isolated from the saliva of participants with low bacterial counts have significant antibacterial capacity compared to that of participants with moderate and high salivary counts. The superior lethality of S. mutans in participants with low salivary counts is likely due to the augmented expression of mutacin- related genes. Copyright © 2016 Elsevier Ltd. All rights reserved.

Community-acquired bacterial meningitis.

PubMed

Costerus, Joost M; Brouwer, Matthijs C; Bijlsma, Merijn W; van de Beek, Diederik

2017-02-01

Bacterial meningitis is a medical emergency and is associated with a high disease burden. We reviewed recent progress in the management of patients with community-acquired bacterial meningitis. The worldwide burden of disease of bacterial meningitis remains high, despite the decreasing incidence following introduction of routine vaccination campaigns. Delay in diagnosis and treatment remain major concerns in the management of acute bacterial meningitis. European Society of Clinical Microbiology and Infectious Diseases guidelines strive for a door-to-antibiotic-time less than 1 h. Polymerase chain reaction (PCR) has emerged as an important diagnostic tool to identify the causative organism. Point-of-care tests using fast multiplex PCR have been developed, but additional value has not been proven. Although anecdotal observations advocate pressure-based management, a randomized controlled trial will need to be performed first to determine efficacy and safety of such an aggressive treatment approach. Adjunctive dexamethasone remains the only adjunctive therapy with proven efficacy. The incidence of bacterial meningitis has been decreasing after the implementation of effective vaccines. Treatment should be administered as soon as possible and time to treatment should not exceed 1 h.

Irradiation treatment of minimally processed carrots for ensuring microbiological safety

NASA Astrophysics Data System (ADS)

Ashraf Chaudry, Muhammad; Bibi, Nizakat; Khan, Misal; Khan, Maazullah; Badshah, Amal; Jamil Qureshi, Muhammad

2004-09-01

Minimally processed fruits and vegetables are very common in developed countries and are gaining popularity in developing countries due to their convenience and freshness. However, minimally processing may result in undesirable changes in colour, taste and appearance due to the transfer of microbes from skin to the flesh. Irradiation is a well-known technology for elimination of microbial contamination. Food irradiation has been approved by 50 countries and is being applied commercially in USA. The purpose of this study was to evaluate the effect of irradiation on the quality of minimally processed carrots. Fresh carrots were peeled, sliced and PE packaged. The samples were irradiated (0, 0.5, 1.0, 2.0, 2.5, 3.0 kGy) and stored at 5°C for 2 weeks. The samples were analyzed for hardness, organoleptic acceptance and microbial load at 0, 7th and 15th day. The mean firmness of the control and all irradiated samples remained between 4.31 and 4.42 kg of force, showing no adverse effect of radiation dose. The effect of storage (2 weeks) was significant ( P< 0.05) with values ranging between 4.28 and 4.39 kg of force. The total bacterial counts at 5°C for non-irradiated and 0.5 kGy irradiated samples were 6.3×10 5 cfu/g, 3.0×10 2 and few colonies(>10) in all other irradiated samples(1.0, 2.0, 2.5 and 3.0 kGy) after 2 weeks storage. No coliform or E. coli were detected in any of the samples (radiated or control) immediately after irradiation and during the entire storage period in minimally processed carrots. A dose of 2.0 kGy completely controlled the fungal and bacterial counts. The irradiated samples (2.0 kGy) were also acceptable sensorially.

Bacteriological monitoring of water reservoirs in oxygen humidifiers: safety of prolonged and multipatient use of prefilled disposable oxygen humidifier bottles.

PubMed

Kobayashi, Nobuharu; Yamazaki, Tsutomu; Maesaki, Shigefumi

2006-03-01

Bacterial colony counts in water specimens from oxygen humidifiers that used reusable water reservoirs were compared with counts in water specimens from humidifiers that used prefilled disposable reservoir bottles to evaluate the effects of prolonged and multipatient use of humidifiers. Bacteria were detected after 1 week of operation in water specimens collected from many humidifiers with reusable reservoirs, but no bacteria were detected in water specimens from disposable bottles for up to 12 weeks during use of the humidifier by multiple patients.

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.

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

Community-acquired bacterial meningitis in adults with cancer or a history of cancer.

PubMed

Costerus, Joost M; Brouwer, Matthijs C; van der Ende, Arie; van de Beek, Diederik

2016-03-01

To study the incidence, clinical presentation, causative bacteria, and outcome of community-acquired bacterial meningitis in adults with cancer. We evaluated incidence and characteristics of patients with cancer included in a nationwide prospective cohort study of adults with community-acquired meningitis performed in the Netherlands from March 1, 2006, to September 31, 2014. All patients underwent a neurologic examination at hospital discharge, and outcome was graded using the Glasgow Outcome Scale. Active cancer was identified in 68 of 1,351 episodes (5%) and a history of cancer in 87 (6%). The annual incidence of community-acquired bacterial meningitis was 2.71-fold (95% confidence interval [CI] 1.68-4.36, p < 0.001) increased for patients with cancer compared to patients without cancer in 2010, and 3.52-fold (95% CI 2.16-5.73, p < 0.001) in 2013. The clinical presentation of bacterial meningitis in patients with cancer compared to patients without cancer was similar. Patients with active cancer presented with lower leukocyte count in blood (12.1 × 10(9) cells/L vs 17.3 × 10(9) cells/L, p < 0.001) and CSF (670 cells/mm(3) vs 2,567 cells/mm(3), p < 0.001) and were more likely to be infected with Listeria monocytogenes (21% vs 5%, p < 0.001) than patients without cancer. Active cancer was identified as an independent risk factor for unfavorable outcome in bacterial meningitis (odds ratio 1.85, 95% CI 1.09-3.13). One of 8 patients with community-bacterial meningitis was identified to have a history of cancer and cancer was considered active in half of these patients. Patients with active cancer present with lower CSF leukocyte counts, are more likely to be infected with L monocytogenes, and are at high risk of unfavorable outcome. © 2016 American Academy of Neurology.

Polyphasic approach for assessing changes in an autochthonous marine bacterial community in the presence of Prestige fuel oil and its biodegradation potential.

PubMed

Jiménez, Núria; Viñas, Marc; Guiu-Aragonés, Cèlia; Bayona, Josep M; Albaigés, Joan; Solanas, Anna M

2011-08-01

A laboratory experiment was conducted to identify key hydrocarbon degraders from a marine oil spill sample (Prestige fuel oil), to ascertain their role in the degradation of different hydrocarbons, and to assess their biodegradation potential for this complex heavy oil. After a 17-month enrichment in weathered fuel, the bacterial community, initially consisting mainly of Methylophaga species, underwent a major selective pressure in favor of obligate hydrocarbonoclastic microorganisms, such as Alcanivorax and Marinobacter spp. and other hydrocarbon-degrading taxa (Thalassospira and Alcaligenes), and showed strong biodegradation potential. This ranged from >99% for all low- and medium-molecular-weight alkanes (C(15)-C(27)) and polycyclic aromatic hydrocarbons (C(0)- to C(2)- naphthalene, anthracene, phenanthrene, dibenzothiophene, and carbazole), to 75-98% for higher molecular-weight alkanes (C(28)-C(40)) and to 55-80% for the C(3) derivatives of tricyclic and tetracyclic polycyclic aromatic hydrocarbons (PAHs) (e.g., C(3)-chrysenes), in 60 days. The numbers of total heterotrophs and of n-alkane-, aliphatic-, and PAH degraders, as well as the structures of these populations, were monitored throughout the biodegradation process. The salinity of the counting medium affects the counts of PAH degraders, while the carbon source (n-hexadecane vs. a mixture of aliphatic hydrocarbons) is a key factor when counting aliphatic degraders. These limitations notwithstanding, some bacterial genera associated with hydrocarbon degradation (mainly belonging to α- and γ-Proteobacteria, including the hydrocarbonoclastic Alcanivorax and Marinobacter) were identified. We conclude that Thalassospira and Roseobacter contribute to the degradation of aliphatic hydrocarbons, whereas Mesorhizobium and Muricauda participate in the degradation of PAHs.

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.

Effect of tannins on growth performance and intestinal ecosystem in weaned piglets.

PubMed

Biagia, Giacomo; Cipollini, Irene; Paulicks, Brigitte R; Roth, Franz X

2010-04-01

Tannins are natural polyphenolic compounds that can reduce digestibility of dietary protein but also display antibacterial effects. The present study investigated, in vitro and in vivo, the effect of different levels of tannins (using a chestnut wood extract containing 75% tannins) on growth performance, intestinal microbiota and wall morphology in piglets. During a 24 h in vitro caecal fermentation, the utilisation of tannins at 0.75, 1.5, 3, and 6 g/l significantly reduced total gas production and concentrations of ammonia and volatile fatty acids and increased viable counts of enterococci and coliforms. When fed to piglets at 1.13, 2.25, and 4.5 g/kg, tannins significantly improved feed efficiency and reduced caecal concentrations of ammonia, iso-butyric, and iso-valeric acid. Viable counts of lactobacilli tended to be increased by tannins in the jejunum, while bacterial caecal counts were not affected. Depth of ileal crypts tended to decrease in piglets fed tannins at 2.25 and 4.5 g/kg. The present study showed that feeding weaned piglets with a tannin-rich wood extract can result in improved feed efficiency and reduction of intestinal bacterial proteolytic reactions. The growth-enhancing effect that tannins had on enterococci and coliforms under in vitro conditions deserves further investigation.

Recovery and diversity of heterotrophic bacteria from chlorinated drinking waters.

PubMed Central

Maki, J S; LaCroix, S J; Hopkins, B S; Staley, J T

1986-01-01

Heterotrophic bacteria were enumerated from the Seattle drinking water catchment basins and distribution system. The highest bacterial recoveries were obtained by using a very dilute medium containing 0.01% peptone as the primary carbon source. Other factors favoring high recovery were the use of incubation temperatures close to that of the habitat and an extended incubation (28 days or longer provided the highest counts). Total bacterial counts were determined by using acridine orange staining. With one exception, all acridine orange counts in chlorinated samples were lower than those in prechlorinated reservoir water, indicating that chlorination often reduces the number of acridine orange-detectable bacteria. Source waters had higher diversity index values than did samples examined following chlorination and storage in reservoirs. Shannon index values based upon colony morphology were in excess of 4.0 for prechlorinated source waters, whereas the values for final chlorinated tap waters were lower than 2.9. It is not known whether the reduction in diversity was due solely to chlorination or in part to other factors in the water treatment and distribution system. Based upon the results of this investigation, we provide a list of recommendations for changes in the procedures used for the enumeration of heterotrophic bacteria from drinking waters. Images PMID:3524453

Inclusion of detergent in a cleaning regime and effect on microbial load in livestock housing

PubMed Central

Hancox, L. R.; Le Bon, M.; Dodd, C. E. R.; Mellits, K. H.

2013-01-01

Determining effective cleaning and disinfection regimes of livestock housing is vital to improving the health of resident animals and reducing zoonotic disease. A cleaning regime consisting of scraping, soaking with or without detergent (treatment and control), pressure washing, disinfection and natural drying was applied to multiple pig pens. After each cleaning stage, samples were taken from different materials and enumerated for total aerobic count (TAC) and Enterobacteriaceae (ENT). Soaking with detergent (Blast-Off, Biolink) caused significantly greater reductions of TAC and ENT on metal, and TAC on concrete, compared with control. Disinfection effect (Virkon S, DuPont) was not significantly associated with prior detergent treatment. Disinfection significantly reduced TAC and ENT on concrete and stock board but not on metal. Twenty-four hours after disinfection TAC and ENT on metal and stock board were significantly reduced, but no significant reductions occurred in the subsequent 96 hours. Counts on concrete did not significantly reduce during the entire drying period (120 hours). Detergent and disinfectant have varying bactericidal effects according to the surface and bacterial target; however, both can significantly reduce microbial numbers so should be used during cleaning, with a minimum drying period of 24 hours, to lower bacterial counts effectively. PMID:23839725

Testing a Low Molecular Mass Fraction of a Mushroom (Lentinus edodes) Extract Formulated as an Oral Rinse in a Cohort of Volunteers

PubMed Central

Signoretto, Caterina; Burlacchini, Gloria; Marchi, Anna; Grillenzoni, Marcello; Cavalleri, Giacomo; Ciric, Lena; Lingström, Peter; Pezzati, Elisabetta; Daglia, Maria; Zaura, Egija; Pratten, Jonathan; Spratt, David A.; Wilson, Michael; Canepari, Pietro

2011-01-01

Although foods are considered enhancing factors for dental caries and periodontitis, laboratory researches indicate that several foods and beverages contain components endowed with antimicrobial and antiplaque activities. A low molecular mass (LMM) fraction of an aqueous mushroom extract has been found to exert these activities in in vitro experiments against potential oral pathogens. We therefore conducted a clinical trial in which we tested an LMM fraction of shiitake mushroom extract formulated in a mouthrinse in 30 young volunteers, comparing the results with those obtained in two identical cohorts, one of which received water (placebo) and the other Listerine. Plaque index, gingival index and bacterial counts in plaque samples were determined in all volunteers over the 11 days of the clinical trial. Statistically significant differences (P < 0.05) were obtained for the plaque index on day 12 in subjects treated with mushroom versus placebo, while for the gingival index significant differences were found for both mushroom versus placebo and mushroom versus Listerine. Decreases in total bacterial counts and in counts of specific oral pathogens were observed for both mushroom extract and Listerine in comparison with placebo. The data suggest that a mushroom extract may prove beneficial in controlling dental caries and/or gingivitis/periodontitis. PMID:21912481

Dynamics of microbiological parameters, enzymatic activities and worm biomass production during vermicomposting of effluent treatment plant sludge of bakery industry.

PubMed

Yadav, Anoop; Suthar, S; Garg, V K

2015-10-01

This paper reports the changes in microbial parameters and enzymatic activities during vermicomposting of effluent treatment plant sludge (ETPS) of bakery industry spiked with cow dung (CD) by Eisenia fetida. Six vermibins containing different ratios of ETPS and CD were maintained under controlled laboratory conditions for 15 weeks. Total bacterial and total fungal count increased upto 7th week and declined afterward in all the bins. Maximum bacterial and fungal count was 31.6 CFU × 10(6) g(-1) and 31 CFU × 10(4) g(-1) in 7th week. Maximum dehydrogenase activity was 1921 μg TPF g(-1) h(-1) in 9th week in 100 % CD containing vermibin, whereas maximum urease activity was 1208 μg NH4 (-)N g(-1) h(-1) in 3rd week in 100 % CD containing vermibin. The enzyme activity and microbial counts were lesser in ETPS containing vermibins than control (100 % CD). The growth and fecundity of the worms in different vermibins were also investigated. The results showed that initially biomass and fecundity of the worms increased but decreased at the later stages due to non-availability of the palatable feed. This showed that quality and palatability of food directly affect biological parameters of the system.

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.

Inducible Defenses Stay Up Late: Temporal Patterns of Immune Gene Expression in Tenebrio molitor

PubMed Central

Johnston, Paul R; Makarova, Olga; Rolff, Jens

2014-01-01

The course of microbial infection in insects is shaped by a two-stage process of immune defense. Constitutive defenses, such as engulfment and melanization, act immediately and are followed by inducible defenses, archetypically the production of antimicrobial peptides, which eliminate or suppress the remaining microbes. By applying RNAseq across a 7-day time course, we sought to characterize the long-lasting immune response to bacterial challenge in the mealworm beetle Tenebrio molitor, a model for the biochemistry of insect immunity and persistent bacterial infection. By annotating a hybrid de novo assembly of RNAseq data, we were able to identify putative orthologs for the majority of components of the conserved insect immune system. Compared with Tribolium castaneum, the most closely related species with a reference genome sequence and a manually curated immune system annotation, the T. molitor immune gene count was lower, with lineage-specific expansions of genes encoding serine proteases and their countervailing inhibitors accounting for the majority of the deficit. Quantitative mapping of RNAseq reads to the reference assembly showed that expression of genes with predicted functions in cellular immunity, wound healing, melanization, and the production of reactive oxygen species was transiently induced immediately after immune challenge. In contrast, expression of genes encoding antimicrobial peptides or components of the Toll signaling pathway and iron sequestration response remained elevated for at least 7 days. Numerous genes involved in metabolism and nutrient storage were repressed, indicating a possible cost of immune induction. Strikingly, the expression of almost all antibacterial peptides followed the same pattern of long-lasting induction, regardless of their spectra of activity, signaling possible interactive roles in vivo. PMID:24318927

Inducible defenses stay up late: temporal patterns of immune gene expression in Tenebrio molitor.

PubMed

Johnston, Paul R; Makarova, Olga; Rolff, Jens

2013-12-06

The course of microbial infection in insects is shaped by a two-stage process of immune defense. Constitutive defenses, such as engulfment and melanization, act immediately and are followed by inducible defenses, archetypically the production of antimicrobial peptides, which eliminate or suppress the remaining microbes. By applying RNAseq across a 7-day time course, we sought to characterize the long-lasting immune response to bacterial challenge in the mealworm beetle Tenebrio molitor, a model for the biochemistry of insect immunity and persistent bacterial infection. By annotating a hybrid de novo assembly of RNAseq data, we were able to identify putative orthologs for the majority of components of the conserved insect immune system. Compared with Tribolium castaneum, the most closely related species with a reference genome sequence and a manually curated immune system annotation, the T. molitor immune gene count was lower, with lineage-specific expansions of genes encoding serine proteases and their countervailing inhibitors accounting for the majority of the deficit. Quantitative mapping of RNAseq reads to the reference assembly showed that expression of genes with predicted functions in cellular immunity, wound healing, melanization, and the production of reactive oxygen species was transiently induced immediately after immune challenge. In contrast, expression of genes encoding antimicrobial peptides or components of the Toll signaling pathway and iron sequestration response remained elevated for at least 7 days. Numerous genes involved in metabolism and nutrient storage were repressed, indicating a possible cost of immune induction. Strikingly, the expression of almost all antibacterial peptides followed the same pattern of long-lasting induction, regardless of their spectra of activity, signaling possible interactive roles in vivo. Copyright © 2014 Johnston et al.

Effect of Lactobacillus johnsonii CRL1647 on different parameters of honeybee colonies and bacterial populations of the bee gut.

PubMed

Audisio, M C; Sabaté, D C; Benítez-Ahrendts, M R

2015-01-01

Lactobacillus johnsonii CRL1647, isolated from the intestinal tract of a worker-bee in Salta, Argentina, was delivered to Apis mellifera L. honey bee colonies according to two different administration schedules: 1×10(5) cfu/ml every 15 days (2011) or monthly (2012). The effect of each treatment on the bee-colony performance was monitored by measuring honey production, and the prevalence of varroasis and nosemosis. Worker bees from each assay were randomly captured 3 days after administration and assayed for the following intestinal culturable and defined bacterial populations: total aerobic microorganisms, Bacillus spp. spores, Lactobacillus spp., Enterococcus spp. and enterobacteria. Interestingly, both treatments generated a similar increase in honey production in treated colonies compared to controls: 36.8% (every 15 days) and 36.3% (monthly). Nosema index always exhibited a reduction when lactobacilli were administered; in turn, Varroa incidence was lower when the lactobacilli were administered once a month. Moreover, the administration of L. johnsonii CRL1647 every 15 days produced an increase in the total number of aerobic microorganisms and in bacteria belonging to the genera Lactobacillus and Enterococcus; at the same time, a decrease was observed in the number of total spores at the end of the treatment. The number of enterobacteria was constant and remained below that of control hives at the end of the assay. On the other hand, the delivery of lactobacilli once a month only showed an increase in the number of bacteria belonging to the genus Lactobacillus; meanwhile, viable counts of the remaining microorganisms assayed were reduced. Even though it seems that both treatments were similar, those bee colonies that received L. johnsonii CRL1647 every 15 days became so strong that they swarmed.

A model for predicting Xanthomonas arboricola pv. pruni growth as a function of temperature

PubMed Central

Llorente, Isidre; Montesinos, Emilio; Moragrega, Concepció

2017-01-01

A two-step modeling approach was used for predicting the effect of temperature on the growth of Xanthomonas arboricola pv. pruni, causal agent of bacterial spot disease of stone fruit. The in vitro growth of seven strains was monitored at temperatures from 5 to 35°C with a Bioscreen C system, and a calibrating equation was generated for converting optical densities to viable counts. In primary modeling, Baranyi, Buchanan, and modified Gompertz equations were fitted to viable count growth curves over the entire temperature range. The modified Gompertz model showed the best fit to the data, and it was selected to estimate the bacterial growth parameters at each temperature. Secondary modeling of maximum specific growth rate as a function of temperature was performed by using the Ratkowsky model and its variations. The modified Ratkowsky model showed the best goodness of fit to maximum specific growth rate estimates, and it was validated successfully for the seven strains at four additional temperatures. The model generated in this work will be used for predicting temperature-based Xanthomonas arboricola pv. pruni growth rate and derived potential daily doublings, and included as the inoculum potential component of a bacterial spot of stone fruit disease forecaster. PMID:28493954

Sediment bacterial indicators in an urban shellfishing subestuary of the lower Chesapeake Bay.

PubMed Central

Erkenbrecher, C W

1981-01-01

The objectives of this study were to document the spatial and temporal distributions and compositions of bacteria in the sediments and overlying waters of an important urban shellfishing area in the lower Chesapeake Bay region, the Lynnhaven Estuary. Marked fluctuations were observed in the date of many of the physicochemical parameters and the indicator bacteria. The higher-salinity water and coarser sediment of the inlet site showed lower overall bacterial densities than did the headwater sites, where freshwater runoff and decreased tidal action were characteristic. Densities of benthic indicator bacteria, when expressed on a volumetric basis, were significantly greater than counts in the overlying waters. These counts were indicative of a fecally polluted system and were well above the safe maximum limits for shellfish-growing waters. Significantly fewer total and fecal bacteria were observed in both the water and the sediment during the warm months of May, July, and August. The primary sources of the Lynnhaven's bacterial pollution appeared to be typical of urban and agricultural runoff, although failure of septic tank systems was suspected as a problem in the Lynnhaven's western branch. These results illustrated that sediments in shellfishing areas could serve as a reservoir for high densities of indicator bacteria and that, potentially, pathogens could pose a health hazard. PMID:7294785

Microbial community assessment of mealworm larvae (Tenebrio molitor) and grasshoppers (Locusta migratoria migratorioides) sold for human consumption.

PubMed

Stoops, J; Crauwels, S; Waud, M; Claes, J; Lievens, B; Van Campenhout, L

2016-02-01

In Western countries, the popularity of edible insects as an alternative animal protein source is increasing. Nevertheless, there is a lack of profound insight into the microbial safety and shelf life of living insects sold for human consumption. The purpose of this study was to characterise the microflora of fresh edible mealworm larvae and grasshoppers in a quantitative and qualitative way. Therefore, culture-dependent analyses (the total viable aerobic count, Enterobacteriaceae, lactic acid bacteria, yeasts and moulds, and bacterial endospores) and next-generation sequencing (454amplicon pyrosequencing) were performed. High microbial counts were obtained for both insect species. Different insect batches resulted in quite similar microbial numbers, except for bacterial endospores. However, the bacterial community composition differed between both insect species. The most abundant operational taxonomic unit in mealworm larvae was Propionibacterium. Also members of the genera Haemophilus, Staphylococcus and Clostridium were found. Grasshoppers were mainly dominated by Weissella, Lactococcus and Yersinia/Rahnella. Overall, a variety of potential spoilage bacteria and food pathogens were characterised. The results of this study suggest that a processing step with a microbiocidal effect is required to avoid or minimize risks involved with the consumption of edible insects. Copyright © 2015 Elsevier Ltd. All rights reserved.

Optical Properties of Three Beach Waters: Implications for Predictive Modeling of Enterococci

EPA Science Inventory

Sunlight plays an important role in the inactivation of fecal indicator bacteria in recreational waters. Solar radiation can explain temporal trends in bacterial counts and is commonly used as an explanatory variable in predictive models. Broadband surface radiation provides a ba...

40 CFR 412.2 - General definitions.

Code of Federal Regulations, 2011 CFR

2011-07-01

....23. (c) Fecal coliform means the bacterial count (Parameter 1) at 40 CFR 136.3 in Table 1A, which... Table 1A at 40 CFR 136.3 are defined as follows: (1) Fecal coliform means fecal coliform bacteria. (2) Total coliform means all coliform bacteria. ...

Photothermal inactivation of bacteria on plasmonic nanostructures

NASA Astrophysics Data System (ADS)

Santos, Greggy M.; Ibañez de Santi Ferrara, Felipe; Zhao, Fusheng; Rodrigues, Debora F.; Shih, Wei-Chuan

2016-03-01

Hospital-acquired bacterial infections are frequently associated with the pathogenic biofilms on surfaces of devices and instruments used in medical procedures. The utilization of thermal plasmonic agents is an innovative approach for sterilizing hospital equipment and for in vivo therapeutic treatment of bacterial infection. A photothermal inactivation technique via array of nanoporous gold disks (NPGDs) has been developed by irradiating near infrared (NIR) light onto deposited bacterial cells (Escherichia coli, Bacillus subtilis, Exiguobacterium AT1B) on the surface of metal nanostructure. The physical and photothermal properties of the NPGD substrate were investigated using topographical scanning electron microscopy (SEM) and thermographic infrared imaging. Bacterial viability studies on NPGD substrates irradiated with and without NIR light were evaluated using a fluorescence-based two-component stain assay. The results show that the heat generated from the NPGD substrate promotes high cell death counts (~100%) at short exposure durations (<25 s) even for thermally-resistant bacterial strains. The photothermal effects on NPGD substrate can lead to point-of-care applications.

Growth and histamine formation of Morganella morganii in determining the safety and quality of inoculated and uninoculated bluefish (Pomatomus saltatrix).

PubMed

Lorca, T A; Gingerich, T M; Pierson, M D; Flick, G J; Hackney, C R; Sumner, S S

2001-12-01

The objective of this study was to determine the effect of normal microflora and Morganella morganii on histamine formation and olfactory acceptability in raw bluefish under controlled storage conditions. Fillets inoculated with and without M. morganii were stored at 5, 10, and 15 degrees C for 7 days. Microbial isolates from surface swabs were identified and screened for histidine decarboxylase activity. Olfactory acceptance was performed by an informal sensory panel. Histamine levels were quantified using high-performance liquid chromatography and fluorescence detection. While olfactory acceptance decreased, histamine concentration and bacterial counts increased. Storage temperature had a significant effect on histamine levels, bacterial counts, and olfactory acceptance of the bluefish. Inoculation with M. morganii had a positive significant effect on histamine formation for bluefish held at 10 and 15 degrees C (P < 0.0001). The results of the study will serve in supporting U.S. Food and Drug Administration (FDA) regulations regarding guidance and hazard levels of histamine in fresh bluefish.

Analysis of low-marbled Hanwoo cow meat aged with different dry-aging methods

PubMed Central

Lee, Hyun Jung; Choe, Juhui; Kim, Kwan Tae; Oh, Jungmin; Lee, Da Gyeom; Kwon, Ki Moon; Choi, Yang Il; Jo, Cheorun

2017-01-01

Objective Different dry-aging methods [traditional dry-aging (TD), simplified dry-aging (SD), and SD in an aging bag (SDB)] were compared to investigate the possible use of SD and/or SDB in practical situations. Methods Sirloins from 48 Hanwoo cows were frozen (Control, 2 days postmortem) or dry-aged for 28 days using the different aging methods and analyzed for chemical composition, total aerobic bacterial count, shear force, inosine 5′-monophosphate (IMP) and free amino acid content, and sensory properties. Results The difference in chemical composition, total aerobic bacterial count, shear force, IMP, and total free amino acid content were negligible among the 3 dry-aged groups. The SD and SDB showed statistically similar tenderness, flavor, and overall acceptability relative to TD. However, SDB had a relatively higher saleable yield. Conclusion Both SD and SDB can successfully substitute for TD. However, SDB would be the best option for simplified dry-aging of low-marbled beef with a relatively high saleable yield. PMID:28728384

Microbial enhancement of compost extracts based on cattle rumen content compost - characterisation of a system.

PubMed

Shrestha, Karuna; Shrestha, Pramod; Walsh, Kerry B; Harrower, Keith M; Midmore, David J

2011-09-01

Microbially enhanced compost extracts ('compost tea') are being used in commercial agriculture as a source of nutrients and for their perceived benefit to soil microbiology, including plant disease suppression. Rumen content material is a waste of cattle abattoirs, which can be value-added by conversion to compost and 'compost tea'. A system for compost extraction and microbial enhancement was characterised. Molasses amendment increased bacterial count 10-fold, while amendment based on molasses and 'fish and kelp hydrolysate' increased fungal count 10-fold. Compost extract incubated at 1:10 (w/v) dilution showed the highest microbial load, activity and humic/fulvic acid content compared to other dilutions. Aeration increased the extraction efficiency of soluble metabolites, and microbial growth rate, as did extraction of compost without the use of a constraining bag. A protocol of 1:10 dilution and aerated incubation with kelp and molasses amendments is recommended to optimise microbial load and fungal-to-bacterial ratio for this inoculum source. Copyright © 2011 Elsevier Ltd. All rights reserved.

Analysis of low-marbled Hanwoo cow meat aged with different dry-aging methods.

PubMed

Lee, Hyun Jung; Choe, Juhui; Kim, Kwan Tae; Oh, Jungmin; Lee, Da Gyeom; Kwon, Ki Moon; Choi, Yang Il; Jo, Cheorun

2017-12-01

Different dry-aging methods [traditional dry-aging (TD), simplified dry-aging (SD), and SD in an aging bag (SDB)] were compared to investigate the possible use of SD and/or SDB in practical situations. Sirloins from 48 Hanwoo cows were frozen (Control, 2 days postmortem) or dry-aged for 28 days using the different aging methods and analyzed for chemical composition, total aerobic bacterial count, shear force, inosine 5'-monophosphate (IMP) and free amino acid content, and sensory properties. The difference in chemical composition, total aerobic bacterial count, shear force, IMP, and total free amino acid content were negligible among the 3 dry-aged groups. The SD and SDB showed statistically similar tenderness, flavor, and overall acceptability relative to TD. However, SDB had a relatively higher saleable yield. Both SD and SDB can successfully substitute for TD. However, SDB would be the best option for simplified dry-aging of low-marbled beef with a relatively high saleable yield.

Childhood meningitis in the conjugate vaccine era: a prospective cohort study.

PubMed

Sadarangani, Manish; Willis, Louise; Kadambari, Seilesh; Gormley, Stuart; Young, Zoe; Beckley, Rebecca; Gantlett, Katherine; Orf, Katharine; Blakey, Sarah; Martin, Natalie G; Kelly, Dominic F; Heath, Paul T; Nadel, Simon; Pollard, Andrew J

2015-03-01

Bacterial conjugate vaccines have dramatically changed the epidemiology of childhood meningitis; viral causes are increasingly predominant, but the current UK epidemiology is unknown. This prospective study recruited children under 16 years of age admitted to 3 UK hospitals with suspected meningitis. 70/388 children had meningitis-13 bacterial, 26 viral and 29 with no pathogen identified. Group B Streptococcus was the most common bacterial pathogen. Infants under 3 months of age with bacterial meningitis were more likely to have a reduced Glasgow Coma Score and respiratory distress than those with viral meningitis or other infections. There were no discriminatory clinical features in older children. Cerebrospinal fluid (CSF) white blood cell count and plasma C-reactive protein at all ages, and CSF protein in infants <3 months of age, distinguished between bacterial meningitis and viral meningitis or other infections. Improved diagnosis of non-bacterial meningitis is urgently needed to reduce antibiotic use and hospital stay. Published by the BMJ Publishing Group Limited. For permission to use (where not already granted under a licence) please go to http://group.bmj.com/group/rights-licensing/permissions.

Influence of zinc on bacterial populations and their proteolytic enzyme activities in freshwater environments: a cross-site comparison.

PubMed

Rasmussen, Lauren; Olapade, Ola A

2016-04-01

Temporal responses of indigenous bacterial populations and proteolytic enzyme (i.e., aminopeptidase) activities in the bacterioplankton assemblages from 3 separate freshwater environments were examined after exposure to various zinc (Zn) concentrations under controlled microcosm conditions. Zn concentrations (ranging from 0 to 10 μmol/L) were added to water samples collected from the Kalamazoo River, Rice Creek, and Huron River and examined for bacterial abundance and aminopeptidase activities at various time intervals over a 48 h incubation period in the dark. The results showed that the Zn concentrations did not significantly influence total bacterial counts directly; however, aminopeptidase activities varied significantly to increasing zinc treatments over time. Also, analysis of variance and linear regression analyses revealed significant positive relationships between bacterial numbers and their hydrolytic enzyme activities, suggesting that both probably co-vary with increasing Zn concentrations in aquatic systems. The results from this study serve as additional evidence of the ecological role of Zn as an extracellular peptidase cofactor on the dynamics of bacterial assemblages in aquatic environments.

Seasonal variability in airborne bacterial communities at a high elevation site and their relationship to other air studies and to potential sources

NASA Astrophysics Data System (ADS)

Bowers, R. M.; Mccubbin, I. B.; Hallar, A. G.; Fierer, N.

2012-12-01

Airborne bacteria are a large component of the near-surface atmospheric aerosol; however we know surprisingly little about their spatiotemporal dynamics and even less about their distributions at high-elevation. With this work, we describe seasonal shifts in bacterial abundances, total particle abundances, and bacterial community structure at a high-elevation research station located in Colorado, USA. In addition, we describe the unique composition of these high-elevation airborne bacterial communities as compared to the bacteria commonly observed throughout the lower elevation atmosphere as well as bacteria common to major sources such as leaf surfaces, soils, water bodies and various other surfaces. To address these knowledge gaps, we collected aerosol samples on the rooftop of Storm Peak Laboratory (3200 m ASL) over the course of 2-3 week periods during each of the four calendar seasons. Total bacterial abundances were assessed via flow cytometry, total particle abundances were calculated with an aerodynamic particle sizer, and bacterial communities were characterized using a high-throughput barcoded DNA sequencing approach. The airborne bacterial communities at Storm Peak Lab were then used in a meta-analysis comparing Storm Peak bacteria to other near-surface (lower elevation) bacterial communities and to the communities of likely source environments. Bacterial abundances varied by season, which was similar but not identical to the changes in total particle abundances across the same sampling period. Airborne bacterial community structure varied significantly by season, with the summer communities being the most distinct. Season specific bacterial groups were identified, suggesting that a large proportion of the airborne community may be derived from nearby sources. However following a multi-environment meta-analysis using several air and source derived bacterial community datasets, the high-elevation air communities were the most distinct as compared to the other airborne communities used in the analysis. Furthermore, a very low proportion of the Storm Peak airborne community could be explained by the source environments used in the meta-analysis, suggesting a unique airborne community at high-elevation. High-alpine bacterial communities appear to make up a large fraction of the total atmospheric aerosol, however the different seasonal patterns between bacterial counts and total particle counts suggest that distinct factors control the quantities of different particles making it into the atmosphere. Furthermore, the characteristics of local terrestrial sources that undergo seasonal cycles seem to have a large influence on the airborne communities, but these sources could not explain the occurrence of all airborne bacterial taxa. As airborne bacteria are more commonly being recognized as a ubiquitous component of the atmosphere, a better understanding of their temporal dynamics in the high-alpine environment may give us insight into their many potential roles in atmospheric dynamics, free troposphere atmospheric dispersal patterns, and their role in human and environmental health.

Assessment of pollution impact on biological activity and structure of seabed bacterial communities in the Port of Livorno (Italy).

PubMed

Iannelli, Renato; Bianchi, Veronica; Macci, Cristina; Peruzzi, Eleonora; Chiellini, Carolina; Petroni, Giulio; Masciandaro, Grazia

2012-06-01

The main objective of this study was to assess the impact of pollution on seabed bacterial diversity, structure and activity in the Port of Livorno. Samples of seabed sediments taken from five selected sites within the port were subjected to chemical analyses, enzymatic activity detection, bacterial count and biomolecular analysis. Five different statistics were used to correlate the level of contamination with the detected biological indicators. The results showed that the port is mainly contaminated by variable levels of petroleum hydrocarbons and heavy metals, which affect the structure and activity of the bacterial population. Irrespective of pollution levels, the bacterial diversity did not diverge significantly among the assessed sites and samples, and no dominance was observed. The type of impact of hydrocarbons and heavy metals was controversial, thus enforcing the supposition that the structure of the bacterial community is mainly driven by the levels of nutrients. The combined use of chemical and biological essays resulted in an in-depth observation and analysis of the existing links between pollution macro-indicators and biological response of seabed bacterial communities. Copyright © 2012 Elsevier B.V. All rights reserved.

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

Effects of soil amendment with different carbon sources and other factors on the bioremediation of an aged PAH-contaminated soil.

PubMed

Teng, Ying; Luo, Yongming; Ping, Lifeng; Zou, Dexun; Li, Zhengao; Christie, Peter

2010-04-01

Carbon supplementation, soil moisture and soil aeration are believed to enhance in situ bioremediation of PAH-contaminated soils by stimulating the growth of indigenous microorganisms. However, the effects of added carbon and nitrogen together with soil moisture and soil aeration on the dissipation of PAHs and on associated microbial counts have yet to be fully assessed. In this study the effects on bioremediation of carbon source, carbon-to-nitrogen ratio, soil moisture and aeration on an aged PAH-contaminated agricultural soil were studied in microcosms over a 90-day period. Additions of starch, glucose and sodium succinate increased soil bacterial and fungal counts and accelerated the dissipation of phenanthrene and benzo(a)pyrene in soil. Decreases in phenanthrene and benzo(a)pyrene concentrations were effective in soil supplemented with glucose and sodium succinate (both 0.2 g C kg(-1) dry soil) and starch (1.0 g C kg(-1) dry soil). The bioremediation effect at a C/N ratio of 10:1 was significantly higher (P < 0.05) than at a C/N of either 25:1 or 40:1. Soil microbial counts and PAH dissipation were lower in the submerged soil but soil aeration increased bacterial and fungal counts, enhanced indigenous microbial metabolic activities, and accelerated the natural degradation of phenanthrene and benzo(a)pyrene. The results suggest that optimizing carbon source, C/N ratio, soil moisture and aeration conditions may be a feasible remediation strategy in certain PAH contaminated soils with large active microbial populations.

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.

Gut Microbiota Composition Before and After Use of Proton Pump Inhibitors.

PubMed

Hojo, Mariko; Asahara, Takashi; Nagahara, Akihito; Takeda, Tsutomu; Matsumoto, Kohei; Ueyama, Hiroya; Matsumoto, Kenshi; Asaoka, Daisuke; Takahashi, Takuya; Nomoto, Koji; Yamashiro, Yuichiro; Watanabe, Sumio

2018-05-24

Recently, problems associated with proton pump inhibitor (PPI) use have begun to surface. PPIs influence the gut microbiota; therefore, PPI use may increase the risk of enteric infections and cause bacterial translocation. In this study, we investigated fecal microbiota composition, fecal organic acid concentrations and pH, and gut bacteria in the blood of the same patients before and after PPI use. Twenty patients with reflux esophagitis based on endoscopic examination received 8 weeks of treatment with PPIs. To analyze fecal microbiota composition and gut bacteria in blood and organic acid concentrations, 16S and 23S rRNA-targeted quantitative RT-PCR and high-performance liquid chromatography were conducted. Lactobacillus species were significantly increased at both 4 and 8 weeks after PPI treatment compared with bacterial counts before treatment (P = 0.011 and P = 0.002, respectively). Among Lactobacillus spp., counts of the L. gasseri subgroup, L. fermentum, the L. reuteri subgroup, and the L. ruminis subgroup were significantly increased at 4 and 8 weeks after treatment compared with counts before treatment. Streptococcus species were also significantly increased at 4 and 8 weeks after PPI treatment compared with counts before treatment (P < 0.01 and P < 0.001, respectively). There was no significant difference in the total organic acid concentrations before and after PPI treatment. Detection rates of bacteria in blood before and after PPI treatment were 22 and 28%, respectively, with no significant differences. Our quantitative RT-PCR results showed that gut dysbiosis was caused by PPI use, corroborating previous results obtained by metagenomic analysis.

A prospective comparison between stabilized glutaraldehyde and chlorhexidine gluconate for preoperative skin antisepsis in dogs.

PubMed

Lambrechts, Nicolaas E; Hurter, Karin; Picard, Jackie A; Goldin, Jeremy P; Thompson, Peter N

2004-01-01

To compare the efficacy of 0.3% stabilized glutaraldehyde and alcohol (SG+A), 0.3% SG and water (SG+W), and 4% chlorhexidine gluconate tincture (CG+A), as skin disinfectants in dogs undergoing ovariohysterectomy. Prospective, blinded clinical study. One hundred and twenty-one dogs. Cutaneous bacterial colony forming units (CFU) from the perioperative site after skin preparation, after antisepsis, and after surgery (incisional and paramedian), were quantified. The influence of high initial bacterial counts (> or =150 CFU) and surgical time on antibacterial efficacy was examined and the proportion of dogs from which Staphylococcus intermedius was cultured, determined. Perioperative skin reactions and wound infections were documented. All 3 antiseptic solutions significantly and equally reduced CFU to all post-antisepsis sampling levels irrespective of surgical duration (mean surgical times 151.6, 136.2, and 149.6 minutes for CG+A, SG+A and SG+W, respectively). Median percentage reductions in CFU ranged between 99.3% and 100%. In dogs with initial high counts and disinfected with CG+A and SG+W, the incisional samples had significantly higher counts than the post-antisepsis samples. In the CG+A and SG+W groups, the proportion of post-surgery samples yielding S. intermedius was significantly higher at the incisional than the paramedian sites. Eight mild cutaneous reactions were recorded in equal proportions for the 3 solutions. There were no recorded infections. All 3 preparations had an equal ability to reduce and maintain low CFU counts, with minimal cutaneous reactions. SG solutions are safe and effective preoperative skin antiseptics for elective clean-contaminated surgical procedures.

Analysis of gingival pocket microflora and biochemical blood parameters in dogs suffering from periodontal disease.

PubMed

Polkowska, Izabela; Sobczyńska-Rak, Aleksandra; Gołyńska, Magdalena

2014-01-01

Periodontal diseases in dogs are caused by bacteria colonising the oral cavity. The presence of plaque comprising accumulations of aerobic and anaerobic bacteria leads to the development of periodontitis. Due to the fact that in a large percentage of cases periodontal diseases remain undiagnosed, and consequently untreated, they tend to acquire a chronic character, lead to bacteraemia and negatively impact the health of internal organs. The aim of the present study was to perform a qualitative microbiological analysis of gingival pockets and determine the correlations between selected morphological and biochemical blood parameters and the extent periodontal diseases. Twenty-one dogs treated for periodontal diseases were qualified for the study and subsequently divided into two groups: with 3rd and 4th stage of periodontal disease. Swabs from the patients' gingival pockets were taken for bacteriological testing. Blood was tested for parameters including erythrocyte count, haemoglobin concentration, haematocrit values and leukocyte count. Blood serum was analyzed with respect to the concentrations of alanine transaminase (ALT), aspartate transaminase (AspAT/AST) and urea. The microbiological analysis of gingival pockets indicated the presence of numerous pathogens with a growth tendency in bacterial cultures observed in dogs with advanced-stage periodontal disease. The concentration of biochemical blood markers was significantly higher in dogs with 4th stage of periodontal disease, to compared to the 3rd-stage group. Morphological parameters were not significantly different with the exception of haemoglobin concentration, which was lower in dogs with 4th stage disease. In both groups, elevated leukocyte counts were observed. By conducting a detailed microbiological examination, it is possible to provide a better prognosis, plan adequate treatment and monitor dogs treated for peridontopathy. Copyright © 2014 International Institute of Anticancer Research (Dr. John G. Delinassios), All rights reserved.

Survival of Lactobacillus acidophilus and Bifidobacterium bifidum in ice cream for use as a probiotic food.

PubMed

Hekmat, S; McMahon, D J

1992-06-01

Probiotic ice cream was made by fermenting a standard ice cream mix with Lactobacillus acidophilus and Bifidobacterium bifidum cultures and then freezing the mix in a batch freezer. Survival of the L. acidophilus and B. bifidum, as well as beta-galactosidase activity, was monitored during 17 wk of frozen storage at -29 degrees C. After freezing of the fermented mix, bacterial counts were 1.5 x 10(8) cfu/ml for L. acidophilus and 2.5 x 10(8) cfu/ml for B. bifidum. Seventeen weeks after freezing, these counts had decreased to 4 x 10(6) and 1 x 10(7) cfu/ml, respectively. During the same period, beta-galactosidase activity decreased from 1800 to 1300 units/ml. Probiotic ice cream was prepared at pH 5.0, 5.5, and 6.0 to determine consumer preferences and was compared with standard Utah State University "Aggie" ice cream. All samples were strawberry-flavored and were evaluated by 88 judges. The preferred pH of probiotic ice cream, based on overall acceptance, was pH 5.5. We demonstrated that probiotic ice cream is a suitable vehicle for delivering beneficial microorganisms such as L. acidophilus and B. bifidum to consumers. The bacteria can be grown to high numbers in ice cream mix and remain viable during frozen storage.

Persistence and biodegradation of kerosene in high-arctic intertidal sediment.

PubMed

Røberg, Stian; Stormo, Svein Kristian; Landfald, Bjarne

2007-10-01

A kerosene type hydrocarbon fraction (equivalent to 7 L m(-2)) was added to enclosures in the surface layer of high-arctic intertidal beach sediment. The experimental spill was repeated in two consecutive years in the period July-September. The rate and extent of hydrocarbon removal and the accompanying bacterial response were monitored for 79 days (2002) and 78 days (2003). The bulk of added kerosene, i.e. 94-98%, was lost from the upper 5 cm layer by putatively abiotic processes within 2 days and a residual fraction in the range 0.6-1.2mg per g dry sediment was stably retained. Concomitant addition of oleophilic fertilizer led to higher initial retention, as 24% of the kerosene remained after 2 days in the presence of a modified, cold-climate adapted version of the well-known Inipol EAP 22 bioremediation agent. In these enclosures, which showed an increase in hydrocarbon-degrader counts from 6.5 x 10(3) to 4.1 x 10(7) per g dry sediment within 8 days, a 17% contribution by biodegradation to subsequent hydrocarbon removal was estimated. Stimulation in hydrocarbon-degrader counts in fertilizer-alone control enclosures was indistinguishable from the stimulation observed with both kerosene and fertilizer present, suggesting that the dynamics in numbers of hydrocarbon-degrading bacteria was primarily impacted by the bioremediation agent.

Bayesian Nonparametric Ordination for the Analysis of Microbial Communities.

PubMed

Ren, Boyu; Bacallado, Sergio; Favaro, Stefano; Holmes, Susan; Trippa, Lorenzo

2017-01-01

Human microbiome studies use sequencing technologies to measure the abundance of bacterial species or Operational Taxonomic Units (OTUs) in samples of biological material. Typically the data are organized in contingency tables with OTU counts across heterogeneous biological samples. In the microbial ecology community, ordination methods are frequently used to investigate latent factors or clusters that capture and describe variations of OTU counts across biological samples. It remains important to evaluate how uncertainty in estimates of each biological sample's microbial distribution propagates to ordination analyses, including visualization of clusters and projections of biological samples on low dimensional spaces. We propose a Bayesian analysis for dependent distributions to endow frequently used ordinations with estimates of uncertainty. A Bayesian nonparametric prior for dependent normalized random measures is constructed, which is marginally equivalent to the normalized generalized Gamma process, a well-known prior for nonparametric analyses. In our prior, the dependence and similarity between microbial distributions is represented by latent factors that concentrate in a low dimensional space. We use a shrinkage prior to tune the dimensionality of the latent factors. The resulting posterior samples of model parameters can be used to evaluate uncertainty in analyses routinely applied in microbiome studies. Specifically, by combining them with multivariate data analysis techniques we can visualize credible regions in ecological ordination plots. The characteristics of the proposed model are illustrated through a simulation study and applications in two microbiome datasets.

Oxytetracycline-Resistant Coliforms in Commercial Poultry Products

PubMed Central

Corey, R. Reece; Byrnes, Joseph M.

1963-01-01

The presence of oxytetracycline-resistant bacteria was investigated with commercially frozen chicken thighs and drumsticks. Bacterial flora were surveyed by means of total and coliform counts with Tryptone Glucose Extract Agar and Desoxycholate Agar, respectively. After counting, the Desoxycholate Agar plates were replicated on the same medium containing 25, 50, 75, and 100 ppm of oxytetracycline. Resistant colonies were found on all samples that were replicated. Of 2613 colonies isolated on Desoxycholate Agar, 47.8% grew in the presence of 25 ppm of oxytetracycline. From 50 to 100 ppm, the number of resistant isolates remained essentially the same, near 34%. Of 812 colonies of antibiotic-resistant bacteria identified with dulcitol-lactose-iron-agar, 82.5% were paracolons, 13.7% were pseudomonads, and 3.8% were Escherichia or Aerobacter. Bacteria resistant to oxytetracycline were shown to be present on commercially processed chicken. The origin of the resistance to oxytetracycline was not established; however, since the antibiotic was not used during processing, it appeared that these antibiotic-resistant bacteria arose in the intestines of the chickens as a result of feed which contained antibiotic. This is supported by a comparison with the antibiotic resistance of coliforms from chickens raised on feed both with and without oxytetracycline, for the percentages of resistant colonies are similar in both commercial chicken and chicken raised on feed containing the antibiotic. PMID:14075046

Reduction effect of bacterial counts by preoperative saline lavage of the stomach in performing laparoscopic and endoscopic cooperative surgery.

PubMed

Mori, Hirohito; Kobara, Hideki; Tsushimi, Takaaki; Fujihara, Shintaro; Nishiyama, Noriko; Matsunaga, Tae; Ayaki, Maki; Yachida, Tatsuo; Tani, Joji; Miyoshi, Hisaaki; Morishita, Asahiro; Masaki, Tsutomu

2014-11-14

To investigate the effects of gastric lavage with 2000 mL of saline in laparoscopic and endoscopic cooperative surgery. Twenty two patients who were diagnosed with a gastric gastrointestinal stromal tumor were enrolled. In former term, irrigations of the stomach were conducted whenever it was necessary, not systematically (Non systemic lavage group). In latter term, the stomach was thoroughly cleaned with 2000 mL of saline using an endoscope with a water jet, and Duodenal balloon occlusion was conducted to prevent refluxed bile and pancreatic juice (Systemic lavage+balloon occlusion group). The gastric wall was sprayed with 20 mL of distilled water, and 20 mL of gastric juice was collected in a sterile tube and submitted for culture. 20 mL of ascites was also collected from the laparoscopic ports and submitted for culture. We compared WBC, CRP, BT between two groups, and verify the reduction effect of bacterial counts in Systemic lavage+balloon occlusion group. WBC count before, 1 d after, and 3 d after laparoscopic and endoscopic cooperative surgery (LECS) were 5060 (95%CI: 4250-9640), 12140 (6050-14110), and 6910 (5320-12520) in Non systemic lavage group, 4400 (3660-7620), 8910 (6480-10980), and 5950 (4840-7860) in Systemic lavage+balloon occlusion group. Significant differences between two groups at the day after LECS (P = 0.029) and the 3 d after LECS (P = 0.042). CRP levels in Non systemic lavage group and in Systemic lavage+balloon occlusion group were significantly different at the day after LECS (P = 0.005) and the 3 d after LECS (P = 0.028). BTs (°C) in Non systemic lavage group and in Systemic lavage+balloon occlusion group were also significantly different at the day after LECS (P = 0.004) and the 3 d after LECS (P = 0.006). In a logarithmic comparison, bacterial load before gastric lavage, after lavage, and ascites culture were 6.08 (95%CI: 4.04-6.97), 0.48 (0-0.85), and 0.21 (0-0.56). The bacterial counts before and after gastric lavage were significantly suppressed (P = 0.007), but no significant difference between gastric juice culture after lavage and ascites (P = 0.154). Pre-LECS lavage with 2000 mL of saline exhibited a bacteria-reducing effect equivalent to disinfectants and obtained favorable results in terms of clinical symptoms and data.

Antibiotic Modification of Native Grafts: Improving upon nature's scaffolds

NASA Astrophysics Data System (ADS)

Ketonis, Constantinos

The use of allograft bone in orthopaedics, spine surgery and dentistry is invaluable for helping restore bone defects and promote osteointegration. However, one, and perhaps the most important, problem associated with the use of allograft is infection. It is a devastating complication for patients and physicians alike, and necessitates repeated surgeries, extended treatment and often times results in increased morbidity and poor outcomes. Previous attempts to incorporate antibiotics into allograft by soaking the graft in antibiotic solution have enjoyed limited success in providing adequate protection against bacterial colonization. To overcome problems associated with controlled release systems, I have described a novel chemical modification that allows for the attachment of vancomycin, or other antibiotics, to free amines of allograft bone thus rendering the graft bactericidal over a long time period. This modification, as evaluated by immunohistochemistry, allowed for the uniform and stable attachment of antibiotics to allograft without adversely affecting its potential for incorporation with bone. Modified allograft, placed in the presence of S. aureus, did not allow colonization by bacteria as evaluated by fluorescent imaging, scanning microscopy, and direct bacterial counts. More importantly, inhibition of bacterial colonization resulted in prevention of biofilm formation. Furthermore, I show that the spectrum of activity of the parent antibiotic was maintained, as the construct was not active against E. coli challenges. Comparison of this technology with simple antibiotic incorporation demonstrated that the covalently-coupled antibiotic did not elute from the bone, but rather remained attached and active on the surface for times out to one year, times that are far longer than currently can be achieved with the elution technologies. Despite its potent activity against bacteria, modified bone remained biocompatible allowing attachment of osteoblastic-like cells with no increased toxicity. Furthermore, the antibiotic-modified allograft incorporated well into tibial defects in the rat. Finally, this construct was efficacious in decreasing the severity of infection and host reaction when impacted in an in vivo model of allograft-associated infection. Thus, our proposed modification in surface design serves as a starting point for the development of a new generation of bone grafts that are biologically active at sites of physiological importance.

Inflammatory response in mixed viral-bacterial community-acquired pneumonia.

PubMed

Bello, Salvador; Mincholé, Elisa; Fandos, Sergio; Lasierra, Ana B; Ruiz, María A; Simon, Ana L; Panadero, Carolina; Lapresta, Carlos; Menendez, Rosario; Torres, Antoni

2014-07-29

The role of mixed pneumonia (virus+bacteria) in community-acquired pneumonia (CAP) has been described in recent years. However, it is not known whether the systemic inflammatory profile is different compared to monomicrobial CAP. We wanted to investigate this profile of mixed viral-bacterial infection and to compare it to monomicrobial bacterial or viral CAP. We measured baseline serum procalcitonin (PCT), C reactive protein (CRP), and white blood cell (WBC) count in 171 patients with CAP with definite etiology admitted to a tertiary hospital: 59 (34.5%) bacterial, 66 (39.%) viral and 46 (27%) mixed (viral-bacterial). Serum PCT levels were higher in mixed and bacterial CAP compared to viral CAP. CRP levels were higher in mixed CAP compared to the other groups. CRP was independently associated with mixed CAP. CRP levels below 26 mg/dL were indicative of an etiology other than mixed in 83% of cases, but the positive predictive value was 45%. PCT levels over 2.10 ng/mL had a positive predictive value for bacterial-involved CAP versus viral CAP of 78%, but the negative predictive value was 48%. Mixed CAP has a different inflammatory pattern compared to bacterial or viral CAP. High CRP levels may be useful for clinicians to suspect mixed CAP.

Sealing effect of a polysaccharide nanosheet for murine cecal puncture.

PubMed

Fujie, Toshinori; Kinoshita, Manabu; Shono, Satoshi; Saito, Akihiro; Okamura, Yosuke; Saitoh, Daizoh; Takeoka, Shinji

2010-07-01

Recent developments in nanobiotechnology have led us to develop a method of producing a free-standing polymer nanosheet composed of polysaccharides (ie, polysaccharide nanosheet) with a thickness of tens of nanometers. Owing to its enormous aspect ratio, the polysaccharide nanosheet is semi-absorbent and has a physical adhesive strength 7.5-fold greater than that of conventional films of >1 microm thickness. Herein, we have investigated the therapeutic sealing effect of this polysaccharide nanosheet on murine cecal puncture as a wound dressing material. Murine cecum was punctured and then overlapped with the polysaccharide nanosheet. Thereafter, we evaluated its sealing effect on bacterial peritonitis as well as the protection offered by the polysaccharide nanosheet against bacterial permeability using an in vitro transmembrane assay. The 39-nm-thick polysaccharide nanosheet overlapped tightly the perforated cecum. No adhering agents were required because of the ability of the polysaccharide nanosheet to adhere to the tissue surface by physical adsorption (eg, van der Waals interaction). Sealing the perforated cecum with the polysaccharide nanosheet increased survival rate without postoperative adhesion by comparison with untreated mice (90 vs 30%; P < .01). These data were supported by the improvement in peritonitis related to bacterial counts, white blood cell counts, and the serum tumor necrosis factor level. Moreover, using an in vitro transmembrane assay, we showed that the polysaccharide nanosheet inhibited effectively bacterial penetration. We have demonstrated the potential clinical benefits of the nanosheet-type biomaterial that can be used for repairing a cecal colotomy without chemical bonding agents. Copyright 2010 Mosby, Inc. All rights reserved.

Stenotrophomonas maltophilia: emergence of multidrug-resistant strains during therapy and in an in vitro pharmacodynamic chamber model.

PubMed Central

Garrison, M W; Anderson, D E; Campbell, D M; Carroll, K C; Malone, C L; Anderson, J D; Hollis, R J; Pfaller, M A

1996-01-01

Emergence of Stenotrophomonas maltophilia as a nosocomial pathogen is becoming increasingly apparent. Pleiotropic resistance characterizes S. maltophilia. Furthermore, a slow growth rate and an increased mutation rate generate discordance between in vitro susceptibility testing and clinical outcome. Despite original susceptibility, drug-resistant strains of S. maltophilia are often recovered from patients receiving beta-lactams, quinolones, or aminoglycosides. Given the disparity among various in vitro susceptibility methods, this study incorporated a unique pharmacodynamic model to more accurately characterize the bacterial time-kill curves and mutation rates of four clinical isolates of S. maltophilia following exposure to simulated multidose regimens of ceftazidime, ciprofloxacin, gentamicin, and ticarcillin-clavulanate. Time-kill data demonstrated regrowth of S. maltophilia with all four agents. With the exception of ticarcillin-clavulanate, viable bacterial counts at the end of 24 h exceeded the starting inoculum. Ciprofloxacin only reduced bacterial counts by less than 1.0 log prior to rapid bacterial regrowth. Resistant mutant strains, identical to their parent strain by pulsed-field gel electrophoresis, were observed following exposure to each class of antibiotic. Mutant strains also had distinct susceptibility patterns. These data are consistent with previous reports which suggest that S. maltophilia, despite susceptibility data that imply that the organism is sensitive, develops multiple forms of resistance quickly and against several classes of antimicrobial agents. Standard in vitro susceptibility methods are not completely reliable for detecting resistant S. maltophilia strains; and therefore, interpretation of these results should be done with caution. In vivo studies are needed to determine optimal therapy against S. maltophilia infections. PMID:9124855