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In vitro evaluation of the antimicrobial activity of chlorhexidine and sodium hypochlorite.

The aim of this study was to investigate in vitro the antimicrobial activity of 0.2%, 1%, and 2% chlorhexidine gluconate (CHX gel and CHX liquid), against endodontic pathogens and compare the results with the ones achieved by 0.5%, 1%, 2.5%, 4%, and 5.25% sodium hypochlorite (NaOCl). A broth dilution test was performed, and the timing for irrigants to kill microbial cells was recorded and statistically analyzed. Both 2.0% gel and liquid formulations eliminated Staphylococcus aureus and Candida albicans in 15 seconds, whereas the gel formulation killed Enterococcus faecalis in 1 minute. All tested irrigants eliminated Porphyromonas endodontalis, Porphyromonas gingivalis, and Prevotella intermedia in 15 seconds. The timing required for 1.0% and 2.0% CHX liquid to eliminate all microorganisms was the same required for 5.25% NaOCl. The antimicrobial action is related to type, concentration, and presentation form of the irrigants as well as the microbial susceptibility.

Anti-Infective Agents, Local↗

Removal and inactivation of Cryptosporidium and microbial indicators by a quaternary ammonium chloride (QAC)-treated zeolite in pilot filters.

A set of pilot filters packed with Zeolite filter media treated with a quaternary ammonium chloride (QAC) were evaluated to verify the proof of concept of their combined antimicrobial capabilities. Escherichia coli was removed and inactivated the most (2.83 log), followed by MS-2 (2.75 log), Klebsiella terriena (2.21 log), PRD-1 (1.95 log), Chlorella vulgaris (1.92 log), and Cryptosporidium parvum oocysts (1.78 log). Especially, inactivation of C. parvum oocysts (1.19 log) was higher than physical removal (0.54 log). The data suggest that QAC-treated Zeolite in the pilot filters has microbial inactivation capabilities and may have useful applications in other types of filter media.

Animals↗

The effect of a portable HEPA-filtered body exhaust system on airborne microbial contamination in a conventional operating room.

OBJECTIVE: To study the effect of a portable HEPA-filtered air exhaust system (Stackhouse Freedom Surgical Helmet System) on airborne microbial contamination in a modern conventional operating room. DESIGN AND SETTING: Microbial air sampling was done with a two-stage Anderson sampler at the wound site during 46 total joint replacements. All operations were performed by the same surgeon in the same operating room at a large community hospital. RESULTS: In 18 cases done without air exhaust hoods, the number of bacterial and fungal colony-forming units (CFU) ranged from 0.6 to 11.7 (mean, 3.6). Air sampling during 28 operations with the operating team in air exhaust hoods revealed a mean of 3.6 CFU (range, 0 to 11.4). Bacterial CFU averaged 3.4 without hoods and 3.2 with exhaust hoods. Coagulase-negative staphylococci were the most common isolates (48% of isolates with hood, 55% without hood). No infections occurred. CONCLUSION: We concluded that these air exhaust hoods did not lower airborne microbial contamination detectable with this air sampling method, as compared to standard head cover and mask, in a modern conventional operating room.

Air Microbiology↗

Effects of hydrocarbon enrichment on trichloroethylene biodegradation and microbial populations in finished compost.

This study focused on the capacity of finished compost, often used as packing material in biofiltration units, to support microbial biodegradation of trichloroethylene (TCE). Finished compost was enriched with methane or propane (10% head space) to stimulate cometabolic biodegradation of gaseous TCE. Successful hydrocarbon enrichment, as indicated by rapid depletion of hydrocarbon gas and measurable growth of hydrocarbon-utilizing micro-organisms, occurred within a week. Within batch reactor flasks, approximately 75% of head space TCE (1-40 ppmv) was rapidly sorbed onto compost material. Up to 99% of the remaining head space TCE was removed via biodegradation in compost enriched with either hydrocarbon. Hydrocarbon enrichment with methane or propane corresponded to 10-fold increases in methanotrophic or propanotrophic populations, respectively. Based on growth assessment under different nutritional regimes, there appeared to be complex metabolic interactions within the microbial community in enriched compost. Five separate bacterial cultures were derived from the hydrocarbon-enriched compost and assayed for the ability to degrade TCE.

Actinomyces↗

Plasminogen is a critical host pathogenicity factor for group A streptococcal infection.

Group A streptococci, a common human pathogen, secrete streptokinase, which activates the host's blood clot-dissolving protein, plasminogen. Streptokinase is highly specific for human plasminogen, exhibiting little or no activity against other mammalian species, including mouse. Here, a transgene expressing human plasminogen markedly increased mortality in mice infected with streptococci, and this susceptibility was dependent on bacterial streptokinase expression. Thus, streptokinase is a key pathogenicity factor and the primary determinant of host species specificity for group A streptococcal infection. In addition, local fibrin clot formation may be implicated in host defense against microbial pathogens.

Ancrod↗

Effects of high pressure treatment on volatile profile during ripening of ewe milk cheese.

The effect of high-pressure treatment on the volatile profile of ewe milk cheeses was investigated. Cheeses were submitted to 200, 300, 400 and 500 MPa at 2 stages of ripening (after 1 and 15 d of manufacturing) and volatile compounds were assayed at 15 and 60 d of ripening. High-pressure treatment altered the balance of volatile profile of cheeses, limiting the formation of acids, alcohols, ketones, aldehydes, and sulfur compounds and enhancing the formation of 2,3-butanedione. In general, cheeses pressurized at 15 d of ripening were more similar to untreated cheeses than those treated at 1 d. Cheeses treated at 300 MPa after 1 d of manufacturing were characterized by higher levels of free amino acids, ethanol, ethyl esters, and branched-chain aldehydes, whereas cheeses treated at 500 MPa after 1 d of manufacturing had lower microbial populations, showed the highest abundance of 2,3-butanedione, pyruvaldehyde, and methyl ketones, and the lowest abundance of alcohols.

Animals↗

Evaluation of microbial hazards during processing of Spanish prepared Flamenquín.

Flamenquín is a traditional, prepared, frozen meat product from the south of Spain made with minced pork, chicken, and cooked ham. Since it is a prepared raw meat product some microbial hazards could be associated with the process of making it. Microbiological analyses have been performed throughout the various steps of processing over a 1-year period to evaluate microbial hazards in the commercial process. High levels of microorganisms were observed all through the processing of this product, the mincing and mixing steps being where major microbial contamination was observed. Pathogenic bacteria such as Staphylococcus aureus, Clostridium perfringens, Escherichia coli and Pseudomonas aeruginosa were detected during processing. Raw materials and food handlers were the principal sources of microbial contamination. A modification of processing to include a heating step after mincing and mixing and an improvement in hygiene practices could eliminate the microbial hazards. Both modifications should be noted for the implementation of a hazard analysis of critical control points (HACCP) program in commercial flamenquín processing.

Clostridium perfringens↗

Development, implementation, and analysis of an on-farm food safety program for the production of greenhouse vegetables.

Fresh fruits and vegetables are increasingly recognized as vectors for foodborne illness. Consequently, an on-farm food safety program was developed, implemented, and analyzed for the Ontario Greenhouse Vegetable Growers in Ontario, Canada, during a 2 1/2-year period. This hazard analysis critical control point-based system was designed to reduce the potential of microbial contamination along the entire production and distribution process. Through the use of microbiological testing, on-site visits, and producer surveys, it was determined that the program has increased grower knowledge, understanding, and awareness of microbial risks associated with fresh produce and caused improvements in practices used within the greenhouse and packing sheds.

Agriculture↗

[Mathematical approach to modeling of the treatment of suppurative processes].

Consideration of an inflammation focus as an "open system" provided analogy between microbiological processes in inflamed wounds and in systems of continuous cultivation of microorganisms. Mathematical modeling of such systems is widely used. Some of the methods for the mathematical modeling were applied to chemoprophylaxis and chemotherapy of postoperative wounds. In modeling continuous cultivation of microorganisms it is usually necessary to determine optimal conditions for the maximum yield of their biomass. In modeling of wound treatment the aim was to determine the process parameters providing the minimum biomass. The described simple models showed that there could be certain optimal flow rate of the washing fluid in the aspiration-washing procedure for wound treatment at which the drug was not completely washed out while the growth rate of the microbial population was minimal. Such mathematical models were shown valuable in optimizing the use of bactericidal and bacteriostatic antibiotics.

Anti-Infective Agents, Local↗

Detection and recovery rates achieved using direct plate and enrichment/immunomagnetic separation methods for Escherichia coli O157:H7 in minced beef and on bovine hide.

AIMS: To assess the detection and recovery rates achieved with commonly used cultural methods for the enumeration and recovery of Escherichia coli O157:H7 from minced beef and bovine hide. METHODS AND RESULTS: Minced beef and bovine hide were inoculated with varying concentrations (log(10) 1.58-2.58 CFU g(-1) and log(10) 2.42-4.49 CFU 100 cm(2) respectively) of E. coli O157:H7 and recovered using a direct plate method or an enrichment/immunomagnetic separation (IMS) method and then plated onto SMAC or SMAC-CT in both cases. The direct plate method detected the pathogen consistently from minced beef samples with an average recovery of 69.2-91.2%. From faecal material on the bovine hide the recovery of the pathogen ranged from 1.80 to 64.5% with fresh faeces depending on the inocula while from dried faeces on hide the results ranged from no recovery at all to 25.1%. Enrichment/IMS recovered E. coli O157:H7 at all inocula levels tested in minced beef while the pathogen was only detected consistently at an average inocula level of log(10) 2.73 CFU 100 cm(2) from fresh faeces and log(10) 4.49 CFU 100 cm(2) from dried faeces on bovine hide. CONCLUSIONS: The direct count enumeration method for E. coli O157:H7 underestimated the numbers of pathogens present. The enrichment/IMS procedure consistently detected the pathogen from minced beef but did not always detect E. coli O157:H7 from faeces on bovine hide. SIGNIFICANCE AND IMPACT OF THE STUDY: Overall this study highlights that any microbial data, used in either predictive microbiology or risk assessment, must take account of the sensitivity and associated performance of the methods employed, in order to make an accurate reflection of the true microbiology of the examined sample.

Animals↗

A comparison of hand washing techniques to remove Escherichia coli and caliciviruses under natural or artificial fingernails.

Compared with other parts of the hand, the area beneath fingernails harbors the most microorganisms and is most difficult to clean. Artificial fingernails, which are usually long and polished, reportedly harbor higher microbial populations than natural nails. Hence, the efficacy of different hand washing methods for removing microbes from natural and artificial fingernails was evaluated. Strains of nonpathogenic Escherichia coli JM109 and feline calicivirus (FCV) strain F9 were used as bacterial and viral indicators, respectively. Volunteers with artificial or natural nails were artificially contaminated with ground beef containing E. coli JM109 or artificial feces containing FCV. Volunteers washed their hands with tap water, regular liquid soap, antibacterial liquid soap, alcohol-based hand sanitizer gel, regular liquid soap followed by alcohol gel, or regular liquid soap plus a nailbrush. The greatest reduction of inoculated microbial populations was obtained by washing with liquid soap plus a nailbrush, and the least reduction was obtained by rubbing hands with alcohol gel. Lower but not significantly different (P > 0.05) reductions of E. coli and FCV counts were obtained from beneath artificial than from natural fingernails. However, significantly (P < or = 0.05) higher E. coli and FCV counts were recovered from hands with artificial nails than from natural nails before and after hand washing. In addition, microbial cell numbers were correlated with fingernail length, with greater numbers beneath fingernails with longer nails. These results indicate that best practices for fingernail sanitation of food handlers are to maintain short fingernails and scrub fingernails with soap and a nailbrush when washing hands.

Alcohols↗

A new approach to determine the genetic diversity of viable and active bacteria in aquatic ecosystems.

BACKGROUND: Discrimination among viable, active, and inactive cells in aquatic ecosystems is of great importance to understand which species participate in microbial processes. In this study, a new approach combining flow cytometry (FCM), cell sorting, and molecular analyses was developed to compare the diversity of viable cells determined by different methods with the diversity of total cells and active cells. METHODS: Total bacteria were determined by SYBR-II staining. Viable bacteria were determined in water samples from different sites by plate count techniques and by the direct viable count (DVC) method. Substrate-responsive cells (i.e., DVC(+) cells) were distinguished from nonresponsive cells (i.e., DVC(-) cells) by FCM and sorted. The genetic diversity of the sorted cell fraction was compared with the diversity of the total microbial community and with that of the culturable cell fraction by denaturing gradient gel electrophoresis (DGGE) of polymerase chain reaction (PCR)-amplified 16S rDNA fragments. The same approach was applied to a seawater sample enriched with nutrients. In this case, actively respiring cells (CTC+) were also enumerated by FCM, sorted, and analyzed by DGGE. RESULTS: The diversity of viable cells varied depending on the methods (traditional culture or DVC) used for viability assessment. Some phylotypes detected in the fraction of viable cells were not detectable at the community level (from total DNA). Similar results were found for actively respiring cells. Inversely, some phylotypes found at the community level were not found in viable and active cell-sorted fractions. It suggests that diversity determined at the community level includes nonactive and nonviable cells. CONCLUSION: This new approach allows investigation of the genetic diversity of viable and active cells in aquatic ecosystems. The diversity determined from sorted cells provides relevant ecological information and uncultured organisms can also be detected. New investigations in the field of microbial ecology such as the identification of species able to maintain cellular activity under environmental changes or in the presence of toxic compounds are now possible.

Bacteria↗

Brewers dried yeast as a source of mannan oligosaccharides for weanling pigs.

Brewers dried yeast, a source of mannan oligosaccharides (MOS), was assessed as an alternative to an antimicrobial agent (carbadox) for young pigs in two experiments. The yeast contained 5.2% MOS. Agglutination tests confirmed adsorption of several serovars of E. coli and Salmonella spp. onto the yeast product. In Exp. 1, seven replicates (five pigs per pen) of 22-d-old pigs were fed a nonmedicated basal diet or the basal diet with carbadox (55 mg/kg), yeast (3%), or a combination of 3% yeast and 2% citric acid for 28 d. Carbadox did not improve growth performance. Growth rate and feed intake were depressed (P < 0.05) in pigs fed yeast alone or in combination with acid. Log counts of total coliforms, Escherichia coli, and Clostridium perfringens in feces were not affected by diet, but Bifidobacteria spp. counts were lower (P < 0.05) in pigs fed the yeast + acid diet and lactobacilli counts were higher (P < 0.05) in pigs fed yeast. Fecal pH and VFA concentrations and intestinal morphological traits were not consistently affected by diet. Serum IgG levels were elevated in the yeast + acid (P < 0.01) group. In Exp. 2, the effects of yeast and carbadox additions to the diet on enteric microbial populations in young pigs housed in isolation units were evaluated. Pigs (n = 24) were weaned at 11 d of age (4.1 kg BW) and placed in isolation chambers (two pigs per chamber) equipped with individual air filtering systems and excrement containers. Treatments were a nonmedicated basal diet and the basal diet with 55 mg/kg of carbadox or with 3% yeast. Diets were fed for 29 d, then each pig was orally dosed with approximately 9.5 x 10(8) CFU of E. coli K88. Daily fecal E. coli K88 counts were not different (P > 0.05) among treatments, but fecal shedding of carbadox-resistant coliforms was higher (P < 0.01) during the 9-d period in pigs fed carbadox. Total fecal coliforms were consistently lower throughout the postinoculation period in pigs fed yeast (P < 0.05). Yeast reduced colonization oftotal coliforms in the duodenum,jejunum, cecum, and colon, but it did not have a consistent effect on colonization of E. coli K88. Pigs fed yeast tended (P < 0.10) to have higher serum IgG levels than controls. In these experiments, brewers dried yeast and carbadox had minimal effects on growth, microbial populations, and intestinal health traits of early-weaned pigs, but certain serum immunological traits were enhanced by feeding yeast.

Agglutination Tests↗

Quantitative analysis of antimicrobial effect kinetics in an in vitro dynamic model.

Variants of the available methods for estimating antimicrobial effect kinetics in an in vitro dynamic model were analyzed. Two integral parameters characterizing antimicrobial effect duration (TE) and intensity (IE) are suggested to define and analyze the concentration-effect relationships in these models, irrespective of the method of recording. TE is defined by the time from the moment of antibiotic administration to the movement when the bacterial count again reaches its initial level. IE is defined by the area between the microbial growth curves in the presence and absence of an antibiotic. TE and IE were used to quantify the antimicrobial effects of sisomicin on Pseudomonas aeruginosa 58, Escherichia coli 93, and Klebsiella pneumoniae 5056, simulating the pharmacokinetic profiles of the drugs observed following intramuscular administration in therapeutic doses, including the variability of aminoglycoside concentrations in human blood.

Anti-Bacterial Agents↗

Reduction of endogenous bacteria associated with catfish fillets using the Grovac process.

Fresh catfish (Ictalurus punctatus) fillets are known to be contaminated with a large number of spoilage and pathogenic bacteria. The Grovac method, a new patented (U.S. 5,543,163) process, was evaluated for its efficacy in reducing the number of pathogens and spoilage microorganisms associated with food. This process involves using a processing solution containing ascorbic acid (AA) and sodium chloride (NaCl), vacuum, and tumbling. A total of 51 bacterial isolates were isolated and identified from whole catfish and catfish fillets using both selective and nonselective media, phenotypic tests, and the Vitek identification system. Psychrotrophic foodborne pathogens included: Aeromonas hydrophila, Escherichia coli, Listeria sp., Plesiomonas shigelloides, Proteus sp., Staphylococcus aureus, and Vibrio parahaemolyticus. High aerobic plate counts (2.6 x 10(7) CFU/g) for catfish fillets indicated that fillets were heavily contaminated during processing of catfish. The Grovac process showed that various treatment combinations of AA and NaCl resulted in a 1.2 to 2.3 CFU/g log reduction of microbial counts associated with catfish fillets. The effectiveness of the process may be related to the synergistic effect of tumbling, AA, NaCl, and vacuum. These results suggested that the Grovac process could be used as an alternative processing procedure to reduce microbial populations associated with catfish fillets and may be useful to improve the shelf-life and food safety of the product. Microbiological data from this study will be used for the development of a hazard analysis for the implementation of the hazard analysis critical control point program for processed catfish fillets.

Animals↗

Microbial contamination of brushes used for preoperative shaving.

Microbial contamination of brushes used for preoperative shaving was investigated. Of the 24 brushed examined, 18 were contaminated with 10(6)-10(9) colony forming units (cfu) per brush. Non-fermentative Gram-negative bacilli such as Pseudomonas aeruginosa and Xanthomonas maltophilia, and yeast-like fungi such as Candida parapsilosis were the primary contaminants. The mean bacterial count on the skin after the use of contaminated brushes (having a mean bacterial count 2.2 x 10(8) cfu) in 14 subjects was 4.6 x 10(5) cfu 25 cm-2, which was about 100 times (p less than 0.001) the control level. Contaminated brushes could not be disinfected with 80% ethyl alcohol, 0.1% sodium hypochlorite or 0.5% chlorhexidine. These findings suggest that the use of brushes should be avoided for preoperative shaving with a razor, and that sterile gauze and shaving foam should be used instead of a brush and soap.

Candida↗

Functional effects of the bacterial insecticide Bacillus thuringiensis var. kurstaki on aquatic microbial communities.

Epilithic microbial communities were colonized on leaf disks and exposed to commercial preparations of Bacillus thuringiensis var. kurstaki (Btk) in aquatic microcosms. Responses in terms of microbial respiration, bacterial cell density, protozoan density, and microbial decomposition activity were measured. Test concentrations for treatments with Dipel 64AF and Dipel 8AF in microcosms were the expected environmental concentration (EEC) of 20 IU/ml, 100x the EEC, and 1000x the EEC. Bacterial cell density in the biofilm of leaf disks was significantly increased at concentrations as low as the EEC. There were no concomitant alterations in protozoan density. Microbial respiration was significantly increased, and decomposition activity was significantly decreased, but only at the artificially high concentration of 1000x the EEC. This effect was attributed to the spore-crystal component rather than formulation ingredients. Microbial decomposition of leaf material was also determined in outdoor stream channels treated at concentrations ranging from the EEC to 100x the EEC. Although there tended to be reduced decomposition activity in treated channels, there were no significant differences in mass loss of leaf material between treated and control channels. Various regression, classification, and ordination procedures were applied to the experimental data, and none indicated significant treatment effects. These results from laboratory and controlled field experiments indicate that contamination of watercourses with Btk is unlikely to result in significant adverse effects on microbial community function in terms of detrital decomposition.

Agammaglobulinaemia Tyrosine Kinase↗

Prevention of catheter-related infections by antiseptic bonding.

A novel catheter pretreated with the antiseptics chlorhexidine and silver-sulfadiazine, designed to reduce catheter-related colonization and infection, was tested in both in vitro and in vivo studies. In vitro experiments demonstrated the long-lasting antibacterial properties of this catheter. For the in vivo study a total of 40 rats divided into different experimental groups were used. Colonization rates of both antiseptic bonded (AS) and control (C) catheters were assessed either three (-3) or seven (-7) days after implantation and local challenge using live Staphylococcus epidermidis ATCC 35984 with 10(7) colony-forming units (cfu) per inoculum. At the time of removal, catheters, organ specimens, and blood samples were taken for cultivation. Significant reductions in the magnitude of colonization of the antiseptic catheters by the test organism were observed in all groups. The average number of cfu colonizing control segments exceeded those found on the treated catheter segments by log 3 (C3/AS3) and log 5 (C7/AS7), respectively (C3: 1.2 x 10(6) +/- 4.1 x 10(5) cfu/segment and AS3: 1.8 x 10(3) +/- 6.6 x 10(2); C7: 2.7 x 10(5) +/- 8.6 x 10(4) and AS7: 1.1 +/- 0.7; mean +/- standard error of the mean, all differences between matching groups statistically significant, Wilcoxon rank sum test, P < 0.0001). These data suggest that antiseptic catheters may substantially decrease the magnitude of catheter-related microbial colonization and subsequent catheter-related infections and may offer a more effective alternative to current methods.

Animals↗