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Levels of microbial contamination on surgical instruments.

OBJECTIVE: To ascertain the microbial load and type of organisms on used surgical instruments following standard cleaning, which consisted of the use of a washer sterilizer followed by sonic cleaning. DESIGN: In this prospective experimental study, used surgical instruments were immersed in Peptamin Tween broth, the broth agitated, and then filtered through a 0.45 microm filter. Quantitative cultures were performed, and all microbes were identified by using standard techniques. SETTING: This study was conducted at a 660-bed university hospital. RESULTS: The microbial load remaining on used surgical instruments after cleaning was as follows: 36 (72%) instruments 0 to 10 colony-forming units (CFU), 7 (14%) instruments 11 to 100 CFU, and 7 (14%) instruments > 100 CFU. Organisms contaminating the instruments included coagulase-negative staphylococcus (56%) followed by Bacillus (22%) and diphtheroids (14%). No other microbes were isolated from more than 4% of the instruments. CONCLUSION: Most used nonlumen surgical instruments contain less than 100 CFU of relatively nonpathogenic microorganisms after cleaning. This suggests that new low-temperature sterilization technologies are likely to be highly effective in preventing cross-transmission of infection via nonlumen medical instruments.

Bacteria↗

Potential rates of fermentation in digesta from the gastrointestinal tract of pigs: effect of feeding fermented liquid feed.

Microbial catabolic capacity in digesta from the gastrointestinal tract of pigs fed either dry feed or fermented liquid feed (FLF) was determined with the PhenePlate multisubstrate system. The in vitro technique was modified to analyze the kinetics of substrate catabolism mediated by the standing stock of enzymes (potential rates of fermentation), allowing a quantitative evaluation of the dietary effect on the catabolic capacity of the microbiota. In total, the potential rates of fermentation were significantly reduced in digesta from the large intestine (cecum, P < 0.1; colon, P < 0.01; and rectum, P < 0.0001) of pigs fed FLF compared to pigs fed dry feed. No effect of diet was observed in the stomach (P = 0.71) or the distal part of the small intestine (P = 0.97). The highest rates of fermentation and the most significant effect of diet were observed for readily fermentable carbohydrates like maltose, sucrose, and lactose. Feeding FLF to pigs also led to a reduction in the large intestine of the total counts of anaerobic bacteria in general and lactic acid bacteria specifically, as well as of microbial activity, as determined by the concentration of ATP and short-chain fatty acids. The low-molecular-weight carbohydrates were fermented mainly to lactic acid in the FLF before being fed to the animals. This may have limited microbial nutrient availability in the digesta reaching the large intestine of pigs fed FLF and may have caused the observed reduction in activity and density of the cecal and colonic microbial population. On the other hand, feeding FLF to pigs reduced the viable counts of coliform bacteria (indicator of Escherichia coli and Salmonella spp.) most profoundly in the stomach and the distal part of the small intestine, probably due to the bactericidal effect of lactic acid and low pH. The results presented clearly demonstrate that feeding FLF to pigs had a great impact on the indigenous microbiota, as reflected in bacterial numbers, short-chain fatty acid concentration, and substrate utilization. However, completely different mechanisms may be involved in the proximal and the distal parts of the gastrointestinal tract. The present study illustrates the utility of the PhenePlate system for quantifying the catabolic capacity of the indigenous gastrointestinal tract microbiota.

Animal Feed↗

Is there a risk of cross-infection from laundered reusable bedpads?

The risk of cross-infection from reusable incontinence bedpads was assessed by determining their microbial content after one night's use by incontinent adults and after laundering using the standard foul wash procedure specified by the NHS Executive (NHS E, 1995) (which includes heat disinfection at 71 degrees C for 3 minutes). Measurements were made on a total of 145 bedpads from five different product designs. It was found that effective laundering destroyed all known pathogenic organisms, although some commensal flora were isolated in small numbers (mean 12.2 colony forming units/ml). It is concluded that laundering reusable incontinence pads using the foul wash procedure leaves pads safe for multiple patient reuse with no demonstrable risk of cross-infection.

Adult↗

Effect of superatmospheric oxygen packaging on sensorial quality, spoilage, and Listeria monocytogenes and Aeromonas caviae growth in fresh processed mixed salads.

Atmospheres with O2 levels higher than 70 kPa have recently been suggested as an innovation to modified atmosphere packaging (MAP) for fresh processed vegetables to maintain sensory quality and safety. In the present work, mixed vegetable salad collected from a commercial processing plant and stored with the MAP technique was studied. Two gas mixtures were actively generated by using an initial O2 concentration of 95 kPa and combined with two plastic films. The low-barrier film permeability for O2 was 1,629 mlO2/m2 x 24 h x atm with 30 microm of thickness (Hyplast, Hoogstraten, Belgium) and the O2 permeability of the high-barrier film was 2 mlO2/m2 x 24h x atm with 150 microm of thickness (Euralpack, Wommelgen, Belgium) at 23 degrees C. As control, active conventional MAP with application of 3 to 5 kPa of O2 and 6 to 8 kPa of CO2 was used. Packaged salads were stored up to 8 days at 4 degrees C and at temperatures simulating chilled distribution chain conditions. Microbial safety and sensory quality, as well as the survival of inoculated Listeria monocytogenes and Aeromonas caviae, were monitored. The effect of superatmospheric O2 on the growth of aerobic microflora was variable. Under superatmospheric conditions, lactic acid bacteria and members of Enterobacteriaceae were inhibited. Nevertheless, growth of yeast and A. caviae seem to be stimulated by superatmospheric O2, whereas growth of psychrotrophic bacteria and L monocytogenes was not affected. The overall visual appearance (mainly color) of the mixed vegetable salads was better maintained and the shelf life prolonged when packaged under O2 concentrations greater than 50 kPa.

Aeromonas↗

Effect of single or sequential hot water and lactic acid decontamination treatments on the survival and growth of listeria monocytogenes and spoilage microflora during aerobic storage of fresh beef at 4, 10, and 25 degrees C.

The survival and growth of Listeria monocytogenes and spoilage microflora during storage of fresh beef subjected to different decontamination treatments was studied. Fresh beef inoculated with a five-strain mixture of L. monocytogenes (5.18 log CFU/cm2) was left untreated (control) or was immersed (30 s) in hot water (HW; 75 degrees C), 2% lactic acid (LA; 55 degrees C), hot water followed by lactic acid (HW-LA), or lactic acid followed by hot water (LA-HW) and then stored aerobically at 4, 10, and 25 degrees C for 25, 17, and 5 days, respectively. Initial populations of L. monocytogenes were reduced by 0.82 (HW), 1.43 (LA), 2.73 (HW-LA), and 2.68 (LA-HW) log CFU/cm2. During storage, the pathogen grew at higher rates in HW than in control samples at all storage temperatures. Acid decontamination treatments (LA. HW-LA, and LA-HW) resulted in a weaker inhibition of L. monocytogenes (P < 0.05) at 25 degrees C than at 4 and 10 degrees C. In general, the order of effectiveness of treatments was HW-LA > LA > LA-HW > HW > control at all storage temperatures tested. In untreated samples, the spoilage microflora was dominated by pseudomonads, while lactic acid bacteria, Enterobacteriaceae, and yeasts remained at lower concentrations during storage. Brochothrix thermosphacta was detected periodically in only a limited number of samples. Although decontamination with HW did not affect the above spoilage microbial profile, acid treatments shifted the predominant microflora in the direction of yeasts and gram-positive bacteria (lactic acid bacteria). Overall, the results of the present study indicate that decontamination with LA and combinations of LA and HW could limit growth of L. monocytogenes and inhibit pseudomonads, which are the main spoilage bacteria of fresh beef stored under aerobic conditions. However, to optimize the efficacy of such treatments, they must be applied in the appropriate sequence and followed by effective temperature control.

Animals↗

Method of applying sanitizers and sample preparation affects recovery of native microflora and Salmonella on whole cantaloupe surfaces.

Standardized methods for applying sanitizer treatments to cantaloupes and for recovering surviving native microflora or Salmonella on inoculated cantaloupe after sanitizing are lacking. Accordingly, the objectives of this study were to compare four methods for applying sanitizers (dipping, dipping with rotation, dipping with agitation, and dipping with rubbing) using 200 ppm of chlorine or 5% H2O2, two recovery methods (homogenization of rind plugs in a stomacher or blender), and five selective recovery media for Salmonella. Whole cantaloupes were submerged in a cocktail of five strains of Salmonella (each at approximately 2 x 10(8) CFU/ml) for 10 min and allowed to dry for 1 h inside a biosafety cabinet and stored at 20 degrees C for approximately 23 h before sanitizing. The recovery of Salmonella from whole cantaloupe without sanitizing averaged 5.09 log CFU/cm2 by blending and 4.30 log CFU/cm2 by homogenization in a stomacher for the five selective agar media. Microbial populations (Salmonella or the indigenous aerobic mesophilic bacteria, gram-negative bacteria, lactic acid bacteria, Pseudomonas spp., and yeast and mold) were not significantly (P > 0.05) reduced by treating with water regardless of the treatment method used. Sanitizing with chlorine or H2O2 by dipping, with or without rotation for 2 min, also did not reduce microbial populations. However, populations of all classes of native microflora and Salmonella were significantly (P < 0.05) reduced by sanitizer treatments (2 min) applied with agitation or by rubbing. In general, sanitizer treatments applied by rubbing resulted in greater log reductions (by up to 1.7 log unit) than for treatments applied with agitation. Populations of native microflora and Salmonella recovered from cantaloupe were higher (by up to 1.8 log unit) by blending compared to homogenization in a stomacher. In most instances, selective media used did not differ significantly (P > 0.05) for recovery of Salmonella after washing treatments.

Anti-Infective Agents, Local↗

Prewashing with acidified sodium chlorite reduces pathogenic bacteria in lightly fermented Chinese cabbage.

Efficacy of prewashing with acidified sodium chlorite (ASC) for the sanitation of lightly fermented Chinese cabbage was evaluated. The population of the natural microflora on the cabbage leaves was reduced about 2.0 log CFU/g just after washing with ASC, a significant reduction compared with the control distilled water wash (P < or = 0.05). In the control experiment, viable aerobic bacteria increased gradually when incubated at 10 degrees C; however, ASC-washed cabbage maintained a lower microbial concentration. The treatment of Chinese cabbage with ASC reduced the population of artificially inoculated Escherichia coli O157:H7, Salmonella Enteritidis, Staphylococcus aureus, and Listeria monocytogenes by 2.4 log CFU/g. The sanitation efficacy of ASC was 1.6 log CFU/g higher than that of distilled water washing. The viable cell counts of all pathogenic bacteria tested remained constant during 8 days of storage at 10 degrees C for both washing treatments, with the exception of L. monocytogenes, whose viable cell counts increased gradually with time for both treatments. No significant differences in color, odor, taste, and texture in raw leaves were observed after the ASC wash compared with after the distilled water wash. These results indicate that prewashing with ASC could control bacterial growth in lightly fermented Chinese cabbage without changing the product quality.

Brassica↗

Evaluation of preoperative hospitalization duration in skin flora.

The skin microbial flora of 18 patients was evaluated during prolonged preoperative hospital stay. Five cultures for bacteria and fungi were obtained on different days: on admission one, three and seven days after admission and after skin disinfection with povidone-iodine solution. There was no change in the mean bacterial count from the admission day to seven days after admission. All but one culture obtained following skin disinfection were negative. Pathogenic bacteria were isolated in only one of each of the following culture day: admission, three and seven days after admission. All cultures were negative for yeasts. The findings suggest that the higher rate of wound sepsis observed in patients with long preoperative hospitalization may not be due to bacterial flora change.

Abdomen↗

Bacterial communities and enzyme activities of PAHs polluted soils.

Three soils (i.e. a Belgian soil, B-BT, a German soil, G, and an Italian agricultural soil, I-BT) with different properties and hydrocarbon-pollution history with regard to their potential to degrade phenanthrene were investigated. A chemical and microbiological evaluation of soils was done using measurements of routine chemical properties, bacterial counts and several enzyme activities. The three soils showed different levels of polycyclic aromatic hydrocarbons (PAHs), being their contamination strictly associated to their pollution history. High values of enzyme activities and culturable heterotrophic bacteria were detected in the soil with no or negligible presence of organic pollutants. Genetic diversity of soil samples and enrichment cultures was measured as bands on denaturing gradient gel electrophoresis (DGGE) of amplified 16S rDNA sequences from the soil and enrichment community DNAs. When analysed by Shannon index (H'), the highest genetic biodiversity (H'=2.87) was found in the Belgian soil B-BT with a medium-term exposition to PAHs and the poorest biodiversity (H'=0.85) in the German soil with a long-term exposition to alkanes and PAHs and where absence, or lower levels of enzyme activities were measured. For the Italian agricultural soil I-BT, containing negligible amounts of organic pollutants but the highest Cu content, a Shannon index=2.13 was found. The enrichment of four mixed cultures capable of degrading solid phenanthrene in batch liquid systems was also studied. Phenanthrene degradation rates in batch systems were culture-dependent, and simple (one-slope) and complex (two-slope) kinetic behaviours were observed. The presence of common bands of microbial species in the cultures and in the native soil DNA indicated that those strains could be potential in situ phenanthrene degraders. Consistent with this assumption are the decrease of PAH and phenanthrene contents of Belgian soil B-BT and the isolation of phenanthrene-degrading bacteria. From the fastest phenanthrene-degrading culture C(B-BT), representative strains were identified as Achromobacter xylosoxidans (100%), Methylobacterium sp. (99%), Rhizobium galegae (99%), Rhodococcus aetherovorans (100%), Stenotrophomonas acidaminiphila (100%), Alcaligenes sp. (99%) and Aquamicrobium defluvium (100%). DGGE-profiles of culture C(B-BT) showed bands attributable to Rhodococcus, Achromobacter, Methylobacterium rhizobium, Alcaligenes and Aquamicrobium. The isolation of Rhodococcus aetherovorans and Methylobacterium sp. can be consistent with the hypothesis that different phenanthrene-degrading strategies, cell surface properties, or the presence of xenobiotic-specific membrane carriers could play a role in the uptake/degradation of solid phenanthrene.

Bacteria↗

Predictive modelling of growth and measurement of enzymatic synthesis and activity by a cocktail of Brochothrix thermosphacta.

The possibility was examined of developing a predictive model that combined microbial growth (increase in cellular number) and extracellular enzyme activity of a cocktail of three strains of Brochothrix thermosphacta. Estimations of growth and enzyme activity were made within a three-dimensional matrix of conditions: temperature 2-20 degrees C, pH value 4.0-7.5 and water activity (a(w)) 0.95-0.995. A model which predicted growth based on increases in cell number was constructed. No extracellular lipases were detected, but slight proteolytic reactions were observed. Although it was not possible to model protease activity, the growth model and information relating to enzyme activity will be made freely available in a database on the Internet.

Colony Count, Microbial↗

Characterization of micrococcaceae isolated from salt used for Spanish dry-cured ham.

The microbial flora of salt used in the production of Spanish dry-cured ham was studied. The results indicated that Micrococcaceae constituted the predominant flora. Identification of the 369 isolates belonging to the family Micrococcaceae revealed that 60% belonged to genus Staphylococcus, 25% to Micrococcus, 6% to Kocuria, 5.4% to Dermacoccus and 0.5% to Stomatococcus. The species most often isolated was Staph. xylosus (28.9%), followed by M. lylae (21.4%), Staph. equorum (18.55%) and D. nishinomiyaensis (5.4%). The results indicate that the salt during salting process of dry-cured hams offers an ecosystem suitable for the survival of the staphylococci and micrococci.

Animals↗

Antimicrobial drug residues in milk and meat: causes, concerns, prevalence, regulations, tests, and test performance.

This paper presents a historical review of antimicrobial use in food animals, the causes of residues in meat and milk, the types of residues found, their regulation in Canada, tests used for their detection, and test performance parameters, with an emphasis on immunoassay techniques. The development of residue detection methods began shortly after the introduction of antimicrobials to food animal production in the late 1940s. From initial technical concerns expressed by the dairy industry to the present public health and international trade implications, there has been an ongoing need for reliable, sensitive, and economical methods for the detection of antimicrobial residues in food animal products such as milk and meat. Initially there were microbial growth inhibition tests, followed by more sensitive and specific methods based on receptor binding, immunochemical, and chromatographic principle. An understanding of basic test performance parameters and their implications is essential when choosing an analytical strategy for residue testing. While each test format has its own attributes, none test will meet all the required analytical needs. Therefore the use of a tiered or integrated system employing assays designated for screening and confirmation is necessary to ensure that foods containing violative residues are not introduced into the food chain.

Animals↗

[Metabolic properties of the microbial community in the biofilters using biolog microplates].

It is very important to know the structure and metabolic function of the microbial community in a bioreactor in order to improve its performance. In this study, two biofilters, packed with wood chips and granular activated carbons respectively, were operated for 160 days to treat toluene gas. The metabolic profiles of the microbial communities in the biofilters were monitored using Biolog microplates periodically during the experiments. The metabolic activities of the microorganisms in both biofilters were observed to decrease during long-term operation. According to the results of principle components analysis, the metabolic profiles of the microbial communities did not change much in the former period of the operation, but they changed in the inlet layers on day 103 and changed throughout the filter beds on day 160. The variation of the metabolic profiles in both biofilters showed little difference, which suggested that the packing media had little effect on them during long-term operation. Among the 95 carbon sources in Biolog microplate, carboxylic acids and amino acids were much easier to be utilized by the microorganisms in the biofilters than the other carbon

Air Pollutants↗

The anti-microbial activity of maggot secretions: results of a preliminary study.

The ability of larval secretions to kill or prevent the growth of a range of potentially pathogenic bacteria was investigated in a preliminary laboratory study. Marked anti-microbial activity was detected against Streptococcus A and B and Staphylococcus aureus. Some activity was also detected against Pseudomonas sp. and a clinical isolate of a resistant strain of S. aureus (MRSA). No evidence of inhibition was recorded against Enterococcus or the Gram-negative bacteria Escherichia coli and Proteus. These findings are consistent with clinical observations made by us and others that larvae of the common greenbottle are able to combat clinical infections in a variety of wound types including those caused by antibiotic-resistant strains of bacteria.

Animals↗

A new technique for quantitative bacterial assessment on burn wounds by modified dermabrasion.

Bacterial colonization and invasive bacterial infection is still one of the major problems in the treatment of burn victims. The standard procedures of bacterial monitoring of the burn would are i) swab-culture which is non-invasive but detects bacteria at the very surface and ii) biopsy-culture which gives a more complete view but has the disadvantage of being invasive. Therefore we developed a new technique for examination of microbial colonization of the wound surface. Dermabrasion of the upper layers of the wound was performed using a small rotating carbon-steel disc of defined roughness. The tissue samples obtained were analysed for bacterial growth in different culture media. Results were qualitatively and quantitatively compared with those of standard techniques performed in parallel. Our results show that this new technique is superior to the swab culture in identifying different bacterial species. The results can be compared with the biopsy technique, but has the advantage of being less invasive.

Adolescent↗

Survival of mycobacteria on N95 personal respirators.

OBJECTIVES: The overall aim of this study was to investigate the survival and possible growth of Mycobacterium tuberculosis simulant bacteria on respirator filters. METHODS: Mycobacterium smegmatis was used as a biochemical simulant for M tuberculosis. Bacterial survival was tested on National Institute for Occupational Safety and Health-certified N95 respirators from three manufacturers. The first experiments simulated one-time respirator use and subsequent storage for 1 to 9 days under ideal conditions for the growth of mycobacteria: 37 degrees C and 85% relative humidity. The bacteria were loaded on the respirator filters under three different nutritional conditions: in the absence of nutrients; in the presence of human saliva (simulating conditions when the respirator is worn); and in the presence of nutrient broth (for ideal growth potential). The subsequent experiments simulated respirator wear for 2 hours under medium work-load conditions at a breathing rate of 56 L/min. RESULTS: It was found that M smegmatis did not grow on the tested respirators, even when the respirators were stored at temperature, humidity, and nutrition conditions most favorable for microbial growth. However, these bacteria could survive on respirators for up to 3 days during storage. The culturability of M smegmatis was not affected by airflow that simulated the breathing rate associated with medium work-load conditions for 2 hours. CONCLUSIONS: This study shows that M tuberculosis surrogate bacteria collected on a respirator are not able to grow and are able to survive only in ideal (ie, not clinically relevant) conditions. Based on these experiments, we conclude that M tuberculosis is unlikely ever to become an infectious problem in the air again, once it is removed by a respirator.

Colony Count, Microbial↗

Biogenic amines and sensory changes associated with the microbial flora of Mediterranean gilt-head sea bream (Sparus aurata) stored aerobically at 0, 8, and 15 degrees C.

Changes in the concentrations of tyramine, agmatine, putrescine, cadaverine, spermidine, tryptamine, spermine, histamine, and trimethylamine were studied in parallel with the development of the microbial population during the storage of Mediterranean gilt-head sea bream (Sparus aurata) at three temperatures (0, 8, 15 degrees C). Changes in sensory scores were also recorded. Pseudomonads and H2S-producing bacteria were the dominant microorganisms. Enterobacteriaceae and lactic acid bacteria were also present in the fish microflora. Among the biogenic amines, putrescine and cadaverine were detected when pseudomonads exceeded 10(6) to 10(7) CFU/g. Histamine was produced only in samples stored at 15 degrees C. Tyramine, tryptamine, agmatine, and trimethylamine were absent regardless of the storage temperature.

Aerobiosis↗

[Microbiological and biochemical indices of changes in microbial ecology of rat large intestine under the effect of rifampicin].

Intragastric administration of rifampicin to rats in a dose of 40 mg/kg resulted in decreasing of the contents of grampositive cocci and lactobacilli and increasing of the number of gramnegative aerobic potentially pathogenic bacteria in feces of the experimental animals. It was noted that along with changes in the composition of the fecal microflora after the exposure to the antibiotic there were disorders in feces excretion of ammonia and amino acids such as alanine and glutamic acid as well as lactic, amber, butyric, valeric and alpha-ketoglutaric acids. Reduction of the changed biochemical indices was shown to be slower than that of the routine microbiological indices.

Animals↗