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PubMed2003

The role of visible faecal material as a vehicle for generic Escherichia coli, coliform, and other enterobacteria contaminating poultry carcasses during slaughtering.

AIMS: A comparison of Enterobacteriaceae, coliform and Escherichia coli counts in chicken carcasses with and without visible faecal contamination was conducted to evaluate the role of contamination as a vehicle for generic E. coli, coliform and other enterobacteria contaminating broiler chicken carcasses when processed under routine commercial operations. METHODS AND RESULTS: Samples were removed from the processing line immediately after evisceration, inside-outside shower and chilling for microbiological analysis. After evisceration, mean counts were significantly different only for E. coli (P < or = 0.05) in chicken carcasses with and without visible faecal contamination. While the spray wash practice was not efficient enough for complete removal of the visible contamination from carcasses, leading to microbiological reduction percentages lower than expected, 25 ppm chlorinated water chilling did reduce the contamination level considerably in all samples. CONCLUSIONS: Carcasses with and without visible faecal contamination harboured E. coli and other potentially hazardous enterobacteria. E. coli was the predominant strain isolated in all samples, Enterobacter cloacae being next most frequent. SIGNIFICANCE AND IMPACT OF THE STUDY: The zero tolerance of visible faecal contamination requirement alone is not sufficient to assure safety and to improve the microbial quality of carcasses.

Abattoirs↗
PubMed2004

Mastitis-causing streptococci are important contributors to bacterial counts in raw bulk tank milk.

The objective of this study was to probe the contribution of streptococci to the microbial quality of raw milk. Over a 5-month period, bulk tank milk samples from 48 New York State dairy farms were analyzed qualitatively for bacterial ecology and quantitatively for total bacterial, streptococcal, staphylococcal, and gram-negative bacterial counts. Linear regression analysis was used to determine the contribution of differential counts to total bacterial counts. Streptococci, staphylococci, and gram-negative bacteria accounted for 69, 3, and 3% of total bacterial count variability, respectively. Randomly selected Streptococcus isolates from each bulk tank milk sample were identified to species by means of the API 20 STREP identification system. The most commonly identified streptococcal species were Streptococcus uberis, Aerococcus viridans, and Streptococcus agalactiae, which were detected in 81, 50, and 31% of 48 bulk tank samples, respectively. For five herds, S. uberis isolates from bulk tank milk and individual cows were characterized by PvuII ribotyping. A farm-specific dominant ribotype was found in each bulk tank sample, and that ribotype was isolated from at least one cow within each herd of origin. Bacteriological and strain typing data indicate that control of streptococci, specifically mastitis-causing species, is important for improvement of the microbial quality of raw milk in New York State.

Animals↗
PubMed1997

Distribution and sources of microbial contamination on beef carcasses.

Three beef dressing lines of different capacity (160, 440 and 800 head d(-1)) were investigated with respect to contamination associated with carcass/hide and carcass/faeces contacts, the distribution of microbial contamination on carcasses and the antimicrobial efficacy of cold water carcass washes. Swab samples were taken from up to 17 sites for determination of Aerobic Plate Counts at 37 degrees C (APC 37 degrees C) and Escherichia coli enumeration using the Petrifilm procedure. The three beef dressing systems produced virtually identical patterns of microbial contamination. High contamination was found at those sites associated with opening cuts and/or subject to hide contact during hide removal. Where contamination is intermittent, the use of mean microbial data tended to obscure evidence of faecal or hide contact. Consequently, worst-case results, as represented by the 95th percentile value, were used to identify probable instances and sources of contact contamination. Sites not subject to faecal contamination or hide contact typically had swab sample APC (37 degrees C) values of less than log 2.00 cfu cm(-2) accompanied by the occasional detection of E. coli at levels below log 1.00 cfu cm(-2). Sites contacted by 'clean' hide typically had APC (37 degrees C) counts of log 3.00 cfu cm(-2) or greater accompanied by occasional E. coli counts not exceeding log 2.00 cfu cm(-2). Sites contaminated by direct faecal contact or contact with faecally contaminated hides typically had APC (37 degrees C) counts equal to, or greater than, log 4.00 cfu cm(-2) accompanied by E. coli counts exceeding log 2.00 cfu cm(-2). Cold water carcass washing was ineffective in removing microbial contamination and tended to bring about a posterior to anterior redistribution, resulting in increased counts at forequarter sites.

Abattoirs↗
PubMed2004

Activation of neutrophils and inhibition of the proinflammatory cytokine response by endogenous granulocyte colony-stimulating factor in murine pneumococcal pneumonia.

Granulocyte colony-stimulating factor (G-CSF) is considered to improve host defense during infection, via increased recruitment of and enhanced performance of neutrophils and subsequent inhibition of potentially harmful proinflammatory mediators. The present study sought to determine the role of endogenous G-CSF in host defense against pneumococcal pneumonia. Patients with unilateral community-acquired pneumonia demonstrated elevated concentrations of G-CSF in bronchoalveolar lavage fluid obtained from the infected, but not from the contralateral, site. Treatment of mice with pneumococcal pneumonia with an anti-G-CSF antibody reduced neutrophil counts in lung tissue and diminished CD11b expression on pulmonary neutrophils but increased the lung concentrations of tumor necrosis factor- alpha, interleukin-1 beta, and cytokine-induced neutrophil chemoattractant. Treatment with anti-G-CSF did not influence the outgrowth of pneumococci in lungs, the dissemination of the infection, or survival in murine pneumonia. During pneumococcal pneumonia, G-CSF is produced locally at the site of the infection, where it exerts both pro- and anti-inflammatory effects.

Adult↗
PubMed1998

Microbial evaluation of Alaska salmon caviar.

Microbial quality of pink salmon caviar (ikura) processed at one plant in Alaska during a 30-day season was examined. Ikura (aw = 0.98; pH 6.1) averaged 49% water, 32% protein, 11% fat, 7% ash, and 3% salt. Aerobic plate counts (APCs) ranged from < 10(2)/g to 4.5 x 10(7)/g with increasing APC toward season's end. Coliform counts ranged from < 3/g to 2.4 x 10(3)/g. Escherichia coli, Staphylococcus aureus, yeasts, and molds were not detected. High-APC (10(7)/g) thawed caviar exhibited predominantly lactic acid bacteria; low-APC (10(3)/g) thawed caviar exhibited predominantly gram-negative bacteria. Freezing had little effect on the microbial counts, and shelf life of thawed caviar was 3 to 5 days at 2 degrees C.

Alaska↗
PubMed2003

Environmental surface cleanliness and the potential for contamination during handwashing.

Effective handwashing (including drying) is important in infection control. The ability of the various stages of handwashing to decrease skin-surface microbial counts has been documented. However, an important element, environmental surface cleanliness, and the potential for contamination of hands during the process has not been well studied or quantified. An examination of the adenosine triphosphate (a measure of residual organic soil), bacterial, and staphylococcal load on ward handwash station surfaces, which could be touched during handwashing, is reported. Hand contact surfaces tested consisted of approximately 620 each of: faucet handles, soap dispenser activator mechanisms, and folded paper-towel dispenser exits. Failure rates in excess of benchmark clean values were higher with adenosine triphosphate assays than microbial counts. This could indicate the presence of a higher level of general organic debris (eg, skin cells) as opposed to microbial contamination or could reflect greater assay sensitivity. Faucet handles were more likely to be contaminated and be in excess of benchmark values than paper-towel dispenser exits. However, the latter are likely to be the final surface touched during the handwashing process and overall nearly 20% were above microbiologic benchmark values. Many of the organisms isolated were staphylococci and the results are discussed within the context of microbial cross-contamination and potential pathogen spread.

Adenosine Triphosphate↗
PubMed2006

Phylogenetic diversity and activity of aerobic heterotrophic bacteria from a hypersaline oil-polluted microbial mat.

The diversity and function of aerobic heterotrophic bacteria (AHB) in cyanobacterial mats have been largely overlooked. We used culture-dependent and molecular techniques to explore the species diversity, degradative capacities and functional guilds of AHB in the photic layer (2mm) of an oil-polluted microbial mat from Saudi Arabia. Enrichment isolation was carried out at different salinities (5% and 12%) and temperatures (28 and 45 degrees C) and on various substrates (acetate, glycolate, Spirulina extract and crude oils). Counts of most probable number showed a numerical abundance of AHB in the range of 1.15-8.13x10(6) cellsg(-1) and suggested the presence of halotolerant and thermotolerant populations. Most of the 16S rRNA sequences of the obtained clones and isolates were phylogenetically affiliated to the groups Gammaproteobacteria, Bacteriodetes and Alphaproteobacteria. Groups like Deltaproteobacteria, Verrucomicrobia, Planctomycetes, Spirochaetes, Acidobacteria and Deinococcus-Thermus were only detected by cloning. The strains isolated on acetate and glycolate belonged to the genera Marinobacter, Halomonas, Roseobacter and Rhodobacter whereas the strains enriched on crude oil belonged to Marinobacter and Alcanivorax. Members of the Bacteriodetes group were only enriched on Spirulina extract indicating their specialization in the degradation of cyanobacterial dead cells. The substrate spectra of representative strains showed the ability of all AHB to metabolize cyanobacterial photosynthetic and fermentation products. However, the unique in situ conditions of the mat apparently favored the enrichment of versatile strains that grew on both the cyanobacterial exudates and the hydrocarbons. We conclude that AHB in cyanobacterial mats represent a diverse community that plays an important role in carbon-cycling within microbial mats.

Bacteria, Aerobic↗
PubMed2000

Fungal colonization and biodeterioration of plasticized polyvinyl chloride.

Significant substratum damage can occur when plasticized PVC (pPVC) is colonized by microorganisms. We investigated microbial colonization of pPVC in an in situ, longitudinal study. Pieces of pPVC containing the plasticizers dioctyl phthalate and dioctyl adipate (DOA) were exposed to the atmosphere for up to 2 years. Fungal and bacterial populations were quantified, and colonizing fungi were identified by rRNA gene sequencing and morphological characteristics. Aureobasidium pullulans was the principal colonizing fungus, establishing itself on the pPVC between 25 and 40 weeks of exposure. A group of yeasts and yeast-like fungi, including Rhodotorula aurantiaca and Kluyveromyces spp., established themselves on the pPVC much later (after 80 weeks of exposure). Numerically, these organisms dominated A. pullulans after 95 weeks, with a mean viable count +/- standard error of 1,000 +/- 200 yeast CFU cm(-2), compared to 390 +/- 50 A. pullulans CFU cm(-2). No bacterial colonization was observed. We also used in vitro tests to characterize the deteriogenic properties of fungi isolated from the pPVC. All strains of A. pullulans tested could grow with the intact pPVC formulation as the sole source of carbon, degrade the plasticizer DOA, produce extracellular esterase, and cause weight loss of the substratum during growth in vitro. In contrast, several yeast isolates could not grow on pPVC or degrade DOA. These results suggest that microbial succession may occur during the colonization of pPVC and that A. pullulans is critical to the establishment of a microbial community on pPVC.

Ascomycota↗
PubMed2006

Microbial adhesion to zirconium alloys.

We present data and analyses concerning the adhesion of clinically relevant Staphylococcus aureus, Staphylococcus epidermidis, and Pseudomonas aeruginosa (bacteria) and Candida albicans (yeast) to Zircaloy-2 (Zry-2) and Zircadyne-705 (Zr705) surfaces. These zirconium-based materials are similar to those now being used in total hip and knee replacements. Here we study clinical strains of microbes under shaken and stationary exposure conditions, and their ability to adhere to Zr surfaces having different oxide thicknesses. We use X-ray photoelectron spectroscopy (XPS), scanning electron microscopy (SEM), viable counts, endotoxin assays, and statistical analysis methods, and demonstrate a predictive model for microbial adhesion based on XPS data.

Alloys↗
PubMed1996

Growth of microorganisms in propofol, thiopental, and a 1:1 mixture of propofol and thiopental.

To assess and compare the growth of four microorganisms in solutions of intravenous anesthetics, known quanta of Staphylococcus aureus, Escherichia coli, Pseudomonas aeruginosa, and Candida albicans were inoculated into propofol, thiopental, a 1:1 mixture of propofol and thiopental, and normal saline. All microorganisms were taken from standard stock cultures and incubated for 24 h (48 h for C. albicans). Growth of microorganism in each drug was compared by counting the number of colony forming units (CFUs) grown from a subculture of each inoculated anesthetic at 0, 3, 6, 12, and 24 h. The study shows that propofol strongly supports the growth of E. coli and C. albicans but is bacteriostatic toward S. aureus and weakly bactericidal toward P. aeruginosa. In contrast, both thiopental and the 1:1 mixture of propofol and thiopental behaved differently, exhibiting markedly bactericidal properties toward E. coli, S. aureus, and P. aeruginosa and a bacteriostatic effect on C. albicans. This finding supports recommendations that a strict aseptic technique should be used when handling propofol and that the contents of an ampoule should be used within 6 h of aspirating. The measured high pH of both thiopental and the 1:1 mixture of propofol and thiopental compared to propofol alone suggests pH to be a major factor in determining whether a given drug will support microbial growth.

Anesthetics, Intravenous↗
PubMed2004

A simple method for differential isolation of freely dispersed and particle-associated peat microorganisms.

Differential isolation of physiologically and ecologically diverse microbial groups facilitates evaluation of their activities in the environment. Here we describe a new method for differential isolation of freely dispersed and particle-associated peat microorganisms. The method is based on sequential elution of peat. Two types of natural peat, "young" sphagnum peat and "old" reed-based peat that have significantly different molecular composition as revealed by solid-state 13C NMR, were used for the method development. Various combinations of elution parameters (pH values, surfactant concentration, and sonication) were tested. Protein yield, CFU counts, 16S rDNA gene sequence, and T-RFLP analyses were used as criteria for evaluating the effectiveness of the elution protocols. The optimal method consisted of sequential elutions with (i) pH 6.2, 0.1 M potassium phosphate buffer, (ii) the same buffer amended with 0.01% of Tween 80, and (iii) treatment (ii) amended with sonication for 2 min. This method allowed differential isolation of freely dispersed and particle-associated microorganisms with (i) highest yield, (ii) highest diversity within the each microbial fraction, and (iii) best discrimination between the freely dispersed and particle-associated microbial communities. The method was applied for characterization of peat microbial communities and revealed different taxonomic composition and similar spatial organization of sphagnum and reed-sage peat microorganisms.

Arthrobacter↗
PubMed2005

Phenotypic identification and technological properties of lactic acid bacteria isolated from traditionally processed fish products of the Eastern Himalayas.

Sukako maacha, gnuchi, sidra and sukuti are traditional smoked and sun-dried fish products of the Eastern Himalayan regions of Nepal and India. A total of 40 samples of sukako maacha (14), gnuchi (6), sidra (10) and sukuti (10) were collected and were analysed for microbial load. Population of lactic acid bacteria (LAB) as well as aerobic mesophilic counts ranged from 4.7-8.3 to 5.1-8.5 log cfu g(-1), respectively. A total of 189 strains of LAB were isolated from sukako maacha, gnuchi, sidra and sukuti samples, out of which 171 strains were cocci and 15 strains, were heterofermentative lactobacilli. LAB were identified on the basis of phenotypic characters including API system as Lactococcus lactis subsp. cremoris, Lactococcus lactis subsp. lactis, Lactococcus plantarum, Leuconostoc mesenteroides, Enterococcus faecium, Enterococcus faecalis, Pediococcus pentosaceus and Weissella confusa. LAB strains produced a wide spectrum of enzymes. Some strains of LAB showed antagonistic properties against pathogenic strains. None of the strains produced biogenic amines in the method applied. This paper is the first report on the microbial composition, mostly lactic acid bacteria, of traditionally processed fish products of Eastern Himalayas.

Animals↗
PubMed2005

Effects of selected pharmaceuticals on riverine biofilm communities.

Although pharmaceutical and therapeutic products are widely found in the natural environment, there is limited understanding of their ecological effects. Here we used rotating annular bioreactors to assess the impact of 10 microg.L(-1) of the selected pharmaceuticals ibuprofen, carbamazepine, furosemide, and caffeine on riverine biofilms. After 8 weeks of development, community structure was assessed using in situ microscopic analyses, fluor-conjugated lectin binding, standard plate counts, fluorescent in situ hybridization, carbon utilization spectra, and stable carbon isotope analyses. The biofilm communities varied markedly in architecture although only caffeine treated biofilms were significantly thicker. Cyanobacteria were suppressed by all 4 compounds, whereas the nitrogen containing caffeine, furosemide, and carbamazepine increased algal biomass. Ibuprofen and carbamazepine reduced bacterial biomass, while caffeine and furosemide increased it. Exopolymer content and composition of the biofilms was also influenced. Significant positive and negative effects were observed in carbon utilization spectra. In situ hybridization analyses indicated all treatments significantly decreased the gamma-proteobacterial populations and increased beta-proteobacteria. Ibuprofen in particular increased the alpha-proteobacteria, beta-proteobacteria, cytophaga-flavobacteria, and SRB385 probe positive populations. Caffeine and carbamazepine additions resulted in significant increases in the high GC354c and low GC69a probe positive cells. Live-dead analyses of the biofilms indicated that all treatments influenced the ratio of live-to-dead cells with controls having a ratio of 2.4, carbamazepine and ibuprofen being 3.2 and 3.5, respectively, and furosemide and caffeine being 1.9 and 1.7, respectively. Stable isotope analyses of the biofilms indicated delta 13C values shifted to more negative values relative to control biofilms. This shift may be consistent with proportional loss of cyanobacteria and relative increase in algal biomass rather than incorporation of pharmaceutical carbon into microbial biofilm. Thus, at 10 microg.L(-1) levels pharmaceuticals exhibit both nutrient-like and toxic effects on riverine microbial communities.

Bacteria↗
PubMed2005

In vitro assessment of the immediate and prolonged antimicrobial action of chlorhexidine gel as an endodontic irrigant against Enterococcus faecalis.

OBJECTIVE: The aim of this study was to assess in vitro the antimicrobial activity of 2% chlorhexidine gel against Enterococcus faecalis , comparing it to other endodontic irrigants (2% chlorhexidine liquid and 5.25% sodium hypochlorite). STUDY DESIGN: Eighty roots of human mandibular premolars were prepared by serial instrumentation technique, autoclaved, and contaminated for 7 days with E faecalis monocultures. The roots were then divided into 5 groups according to the irrigant substance used during the standardized biomechanical preparation. In order to evaluate the antimicrobial action of the irrigant substances, 3 microbial samples were taken: initial (before the biomechanical preparation); post-treatment (immediately after the biomechanical preparation), and final (7 days after the biomechanical preparation). The microbiological samples were plated to count the colony-forming units (CFU). RESULTS: The 2% chlorhexidine gel and 2% chlorhexidine liquid significantly reduced the E faecalis CFU in the post-treatment and final microbiological samples. The 5.25% sodium hypochlorite also reduced the E faecalis CFU immediately after the root canal instrumentation, but it was not able to keep the root canal free of detectable E faecalis in the final sample. CONCLUSIONS: The 2% chlorhexidine gluconate (gel and liquid) antimicrobial ability was more effective than 5.25% sodium hypochlorite in keeping low CFU of E faecalis for 7 days after the biomechanical preparation.

Anti-Infective Agents, Local↗
PubMed2003

Development of an adhesive sheet for direct counting of bacteria on solid surfaces.

An adhesive sheet was developed for direct counting of microorganisms on solid surfaces. The sheet consists of a polyurethane film base and water insoluble adhesive. SYBR Green II (for total direct counting) or 6-carboxyfluorescein diacetate (6CFDA) (for fluorescent vital staining) was used for fluorescent microscopy of bacteria collected on the adhesive face of the sheet. Adhesive sheet sampling showed a higher recovery rate for microbial enumeration than conventional swab method or stamp agar. This method is simple, rapid, inexpensive and reproducible.

Adhesives↗
PubMed2003

Nutritional evaluation of egg byproducts in diets for early-weaned pigs.

A total of 272 Cotswold pigs (17 +/- 1 d) were utilized in three experiments to evaluate the nutritive value of spray-dried egg proteins for early-weaned pigs. In all experiments, pigs were stratified by sex and initial BW and then assigned randomly to experimental diets. In Exp. 1, four corn-soybean meal-based diets containing 7% of either spray-dried porcine plasma (SDPP), spray-dried technical albumen (SDTA), SDTA stored at 70 degrees C for 3 d (SDTA-ht), or spray-dried whole egg (SDWE) were assigned to five pens each with four pigs for a 3-wk study period. Average daily gain, ADFI, and gain:feed ratio (G:F) were determined. At the end of wk 3, five pigs per treatment were killed to determine ileal AA and energy digestibilities, as well as Enterobacteriaceae counts. Compared with the SDPP diet, ADG and G:F were lower (P < 0.05) for SDTA-, SDTA-ht- and SDWE-containing diets. Apparent ileal digestibilities of cystine, histidine, isoleucine, methionine, and threonine in the SDPP diet were lower (P < 0.05) than in diets containing spray-dried egg products. Ileal digestible energy content did not differ (P > 0.05) in all diets (3.1 to 3.2 Mcal/kg). Enterobacteriaceae counts were lower in the SDTA-ht diet than in either the SDTA or SDWE diets (P < 0.05). In Exp. 2, the effect of substituting SDPP with varying levels of SDTA was investigated. Diets were randomly assigned to five pens (except for the 100% SDTA diet, which had four pens), each with four pigs. Average daily gain, ADFI, and G:F decreased linearly as the level of SDTA was increased in the diet (P < 0.05). Replacing SDPP with SDTA at 25 or 50% had no effect on pig performance (P > 0.10). In Exp. 3, phase I diets containing 0, 25, or 50% SDTA in place of SDPP (7% of the diet) were each assigned at random to eight pens each with four pigs for a 14-d period, after which all pigs were switched to a common phase II diet lacking both SDPP and SDTA for another 14 d. Average daily feed intake and ADG did not differ among all diets in phase I and II and overall (d 0 to 28). Pigs fed the diet containing 50% SDTA in phase I had lower (P < 0.05) G:F than those fed the SDPP diet. The results indicate that technical albumen can replace 25 to 50% of SDPP in early-weaned pig diets without compromising performance, and further suggest that heat-treated SDTA may affect intestinal microbial population in pigs.

Amino Acids↗
PubMed2006

[Influence of milking technique, milking hygiene and environmental hygiene parameters on the microbial contamination of milking machines].

It was the aim of this study to investigate the effect of various factors of the milking technique, milking hygiene and environment on microbial contamination of the milking machine. In 31 dairy herds, the degree of bacterial contamination was examined by taking swabs at four locations (teat cup liner, claw, short and long milk tube) before the milking procedure was started using a standardized protocol (DIN ISO 6887-1:1999). Furthermore, the total germ count was determined in the first milk entering the bulk tank as well as in the bulk tank milk following milking. For each farm, the quality of the milking process and the condition of the milking machine as well as of various environmental factors were recorded. A subjective evaluation of the status of the milking cluster or other parts of the milking machine ("good" or "moderate-poor") gave more information about bacterial contamination than the determination of age and type of material used. A temperature of the rinsing water of < 42 degrees C increased the contamination with Pseudomonas spp. and coliform bacteria. Milking clusters kept out of the cluster pick-up between milking had a higher risk of microbial contamination. Various methods of teat cleaning before milking or of postmilking teat disinfection did not affect the contamination of the milking machine and the bulk tank milk with environmental bacteria. Furthermore, type of bedding material affected bacterial contamination of milking clusters and bulk tank milk. In conclusion, our results suggest that the microbial contamination of the milking machine is not only influenced by the sanitation pro-

Animals↗