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Microbial contamination of water-soaked cotton gauze and its cause.

Seven in-use cotton gauze samples and three cotton balls soaked in sterile distilled water in canisters were investigated 7 days after they were prepared in hospital. All samples were contaminated with bacteria including 10(6) to 10(7) colony forming units/ml of Pseudomonas aeruginosa. In vitro viability tests using cotton gauze and cotton balls soaked in sterile distilled water revealed rapid proliferation of P. aeruginosa, Serratia marcescens and Candida albicans. Since the cotton gauze and the cotton balls were soaked in water containing nutrients, such as protein and glucose, these materials may be readily contaminated with bacteria including P. aeruginosa. Thus, when using cotton gauze and cotton balls containing water, microbial contamination should be expected.

Candida albicans↗

Effect of nutrients on defined bacterial plaques and Streptococcus mutans C67-1 implantation in a model mouth.

Actinomyces viscosus WVU 627, Streptococcus oralis LPA-1 and Veillonella dispar OMZ 193 were cocultured on teeth in a model mouth for 66 h. Synthetic saliva containing bovine salivary glycoprotein supported bacterial growth, although the delivery of an intermittent nutrient supplement, containing 1% (w/v) glucose or sucrose, gave greater bacterial cell and viable counts. When Streptococcus mutans C67-1 was super-inoculated onto 24-hour mixed plaques, it became established under all regimens, but there was pronounced colonization resistance. With saliva only, the proportion of S. mutans at 66 h was less than 0.5% of the total cultivable microflora. When a glucose supplement was delivered for 1 h every 6 h, S. mutans attained a final proportion of 2.4%. With sucrose, both S. mutans C67-1 and its non-cariogenic glucan-deficient mutant, C67-25, attained similar proportions of 15-20%. These experiments indicate how this model can be used to study the factors influencing colonizing ability and microbial interactions in biofilms under controlled conditions.

Actinomyces↗

Relationship between skin microbial counts and surgical site infection after neurosurgery.

A prospective study was performed to describe the density of bacterial counts on the skin of neurosurgical patients and examine the association between total colony-forming unit (cfu) counts of skin flora at the operative site and surgical site infection (SSI). Two skin cultures were obtained, immediately before and after skin preparation, from the operative sites of 609 neurosurgical patients. SSI surveillance that used Centers for Disease Control/National Nosocomial Infection Surveillance definitions was performed. Predictors for high bacterial counts and SSI among craniotomies were analyzed by means of logistic regression. Neither pre- nor postpreparation counts were associated with SSI. Other SSI risk factors were obesity (relative risk [RR], 2.5), duration of surgery (RR, 1.3 for every additional 30 minutes) and age (RR, 0.7 for each additional 10 years). Duration of skin preparation was not correlated with postpreparation cfu counts. We were unable to detect an association between preoperative bacterial skin counts and SSI.

Adolescent↗

Bacterial contamination of blenderized whole food and commercial enteral tube feedings in the Philippines.

Hospital-prepared tube feedings from four Philippine acute-care hospitals were analysed for microbial contamination. Two feedings were prepared on three separate days at each hospital. The tube feedings were either blended natural whole foods or a reconstituted commercial powdered nutritional product. Samples of each feeding were collected for coliform count and standard plate count at the time of preparation and at 1, 2 and 4 h after preparation after maintenance at room temperature (26-31 degrees C). At the time of preparation, mean coliform and standard plate counts for all samples were 10.3 most probable number per gram (MPN/g) and 7.4x10(4)colony-forming units per gram (cfu/g), respectively. Nine of 24 samples (38%) had coliform counts greater than 10 MPN/g, and 22/24 (92%) samples had standard plate counts greater than 10(3) cfu/g. There were significant increases in mean coliform and standard plate counts over 4 h (P=0.0005 and P=0.008, respectively). At 4 h after preparation, the mean coliform and standard plate counts were 18.2 MPN/g and 2.1x10(5) cfu/g, respectively. At this time, 18/24 (75%) samples had coliform counts greater than 10 MPN/g and 20/24 (83%) samples had standard plate counts greater than 10(5) cfu/g. The results of this study show that the microbial quality of the majority of the hospital-prepared enteral tube feedings analysed were not within published guidelines for safety.

Colony Count, Microbial↗

The direct effect of ultrasound on the extraction of date syrup and its micro-organisms.

In the date syrup industry, date fruits are mixed with a suitable amount of water at a temperature greater than 50 degrees C for about 1 h. This condition is not sufficient for killing the micro-organisms present in the fruit. In addition, Overheating for a long time can damages nutritious materials and also changes the final product's color. Ultrasound was applied for improving the quantity and quality of the extraction, and also to overcome the health problem. In this research, the following variables were examined: date fruit/water ratio, ultrasonic intensity and temperature. The results showed that sonication under the proper conditions can lead to a higher extraction in a shorter time with a better physical quality of the product. Most importantly, the sonication significantly decreased the microbial count in comparison to the classical method. This study also confirmed the presence of anti-microbial substances in date fruit, and that ultrasonic waves can accelerate their effects.

Colony Count, Microbial↗

Comparison of the intestinal bacteria in specific pathogen free mice from different breeders.

Specific pathogen free BALB/c mice from 3 commercial laboratory animal breeders in Japan were compared on the composition of caecal flora revealed by selective and nonselective cultivation as well as direct microscopical observation on smears, and relative caecal weight. Large differences were detected in viable counts of total bacteria and almost all bacterial groups, while direct microscopical counts which consisted mainly of fusiform bacteria were almost equal, resulting in diverse recovery rates among 3 breeders. Eubacterium and spiral shaped organisms were not detected from all breeders. There also existed variations within breeders, especially those in the number of Enterobacteriaceae. Relative caecal weight also differed among breeders, suggesting the participation of variety of caecal bacteria in determining this parameter. As these variations in bacteriological status of animals can influence experimental results, standardization of intestinal microbial flora is required.

Animals↗

Organic acid dipping of catfish fillets: effect on color, microbial load, and Listeria monocytogenes.

Microbiological and color changes of catfish fillets were determined following dip treatment in solutions at 4 degrees C of 2% acetic, citric, hydrochloric, lactic, malic, or tartaric acid. Fillets were inoculated with an eight-strain mixture of Listeria monocytogenes prior to dipping. L. monocytogenes, coliform, and aerobic plate counts and surface pH and Hunter color were measured at 0, 2, 5, and 8 days of storage at 4 degrees C. Acid dipping reduced surface pH and L. monocytogenes, coliform, and aerobic microbial loads. Little microbial proliferation was observed on acid-treated fillets, however, controls had a distinct foul odor and microbial loads in excess of 10(6) CFU/g by day 8. On untreated fillets, L. monocytogenes counts did not increase during storage, perhaps due to competitive inhibition by normal catfish microflora. Hunter color analysis revealed lighter and yellower acid-treated fillets than untreated controls, with malic acid producing the least bleaching. The shelf life of refrigerated fillets increased when fillets were acid dipped. It remains to be established if this enhanced microbial quality also parallels sensory acceptability.

Animals↗

Bioluminescence as a real-time monitor of injury and recovery in Salmonella typhimurium.

Bioluminescence provides a novel tool with which to probe in real time the recovery of microorganisms such as Salmonella typhimurium from sub-lethal injury, a condition of immense importance in the enumeration of microorganisms in food and environmental samples such as from estuarine waters. We describe the application of in vivo bioluminescence to monitor the recovery of S. typhimurium from freeze injury and, in addition, we explore the effect of multiple stress conditions on microbial recovery where we compare bioluminescence with viable count measurement.

Colony Count, Microbial↗

Abundance of polymers degrading microorganisms in a sea-based solid waste disposal site.

In order to assess the degradability of plastics in solid waste disposal landfill sites, microbial populations capable of degrading five kinds of plastic-constituting polymers, poly epsilon-caprolactone (PCL), polylactic acid (PLA), polyethylene glycol (PEG), poly-beta-hydroxybutyrate (PHB) and cellulose acetate (CA), in a sea-based solid waste disposal site were investigated. Enumeration of aerobic and anaerobic polymers-degrading microorganisms (PDMs) was performed against to total 8 leachate samples, which were seasonally collected from the facultative pretreatment pond and the aerated lagoon. Both aerobic and anaerobic PDMs for natural polymers, PHB and CA, were found in all of the samples, while those for chemically-synthesized polymers, PCL, PLA and PEG, could not be always detected. In most cases, the ratios of the PHB- and CA-degraders to the heterotrophic bacterial population were more than 0.1%. On the other hand, the ratios of PCL-, PLA- and PEG-degraders were often much lower. These data indicate that the plastics degradation potential is commonly present in the studied disposal site, and that the degradation potential for plastics composed of chemically-synthesized polymers is inferior to that of natural polymers. Population sizes of the PDMs correlated to those of heterotrophic bacteria, and the counts of aerobic heterotrophic bacteria and PDMs in the aerated lagoon tended to be higher than those of anaerobic ones, indicating that the aeration of the leachate resulted in the activation of growth of whole aerobic microbial community including the PDMs.

Bacteria↗

Bacterial diversity in spent mushroom compost assessed by amplified rDNA restriction analysis and sequencing of cultivated isolates.

Spent mushroom compost (SMC) is the residual by-product of commercial Agaricus spp. cultivation, and it is mainly composed of a thermally treated cereal straw/animal manure mixture colonized by the fungal biomass. Research on the valorization of this material is mainly focusing on its use as soil conditioner and plant fertilizer. An investigation of the bacterial diversity in SMC was performed using molecular techniques in order to reveal the origin of SMC microflora and its potential effect on soil microbial communities after incorporation into agricultural soils. The bacterial population was estimated by the plate count method to a mean of 2.7 10(9) colony forming units (cfu) per g of dry weight, while the numbers of Gram-positive and Gram-negative bacteria were 1.9 10(9) and 4.9 10(8) cfu per g dw respectively as estimated by enumeration on semi-selective media. Fifty bacterial isolates were classified into 14 operational taxonomic units (OTUs) following ARDRA-PCR of the 16S rDNA gene. Sequencing of the 16S rDNA amplicon assigned 12 of the 14 OTUs to Gram-positive bacteria, associated with the genera Bacillus, Paenibacillus, Exiguobacterium, Staphylococcus, Desemzia, Carnobacterium, Brevibacterium, Arthrobacter and Microbacterium of the bacterial divisions Firmicutes and Actinobacteria. Two bacterial groups have phylogenetic links with the genera Comamonas and Sphingobacterium, which belong to beta-Proteobacteria and Bacteroidetes respectively. Two potentially novel bacteria are reported, which are associated with the genera Bacillus and Microbacterium. Most of the bacteria identified are of environmental origin, while strains related to species usually isolated from insects, animal and clinical sources were also detected. It appears that bacterial diversity in SMC is greatly affected by the origin of the initial material, its thermal pasteurization treatment and the potential unintended colonization of the mushroom substrate during the cultivation process.

Agaricus↗

Comparison of a rapid ATP bioluminescence assay and standard plate count methods for assessing microbial contamination of consumers' refrigerators.

The feasibility of using an ATP bioluminescence assay for assessing microbial contamination of home refrigerators was evaluated and compared with the standard culture methods. Samples of refrigerator surfaces were collected from 123 households by swabbing an area of 100 cm2 on three locations in the refrigerator with premoisturized sterile swabs. Microbial contaminations were determined by aerobic plate count (APC; incubated at 35 degrees C for 48 h) and psychrotrophic plate count (PPC; incubated at 7 degrees C for 10 days) on plate count agar. The results were compared to the readings from the microbial ATP (mATP) bioluminescence assay. The correlation coefficient (r) between mATP and PPC (r = 0.851) was slightly higher than that between mATP and APC (r = 0.823). Our results indicated a potential discrepancy in the population of mesophilic and psychrotrophic bacteria in the refrigerator samples. Nevertheless, mATP appeared to be a reliable indication of the average of APC and PPC (r = 0.895). The mATP bioluminescence assay would provide a rapid and convenient test for researchers in field studies to assess microbial contamination in refrigerators.

Adenosine Triphosphate↗

Electrochemical disinfection of drinking water using an activated-carbon-fiber reactor capable of monitoring its microbial fouling.

An electrochemical reactor employing activated carbon fibers (ACF) was constructed for the disinfection of bacteria in drinking water. The application of an alternating potential of 1.0 V and -0.8 V versus a saturated calomel electrode, for disinfecting and desorbing bacteria, enabled reactor operation for 840 h. Drinking water was passed through the reactor in stop/flow mode: 300 ml/min flow for 12 h and no flow for 12 h, alternately. The bacterial cell density in treated water was always been less than 20 cells/ml. It was also found that the formation of biofilm on the ACF reactor caused an increase in current, enabling the self-detection of microbial fouling.

Absorption↗

Pulsed electric fields cause sublethal injury in Escherichia coli.

AIMS: The objective was to investigate the occurrence of sublethal injury in Escherichia coli by pulsed electric fields (PEF) at different pH values. METHODS AND RESULTS: The occurrence of sublethal injury in PEF-treated E. coli cells depended on the pH of the treatment medium. Whereas a slight sublethal injury was detected at pH 7, 99.95% of survivors were injured when cells were treated at pH 4 for 400 micros at 19 kV. The PEF-injured cells were progressively inactivated by a subsequent holding at pH 4. CONCLUSIONS: PEF cause sublethal injury in E. coli. The measurement of sublethal injury using a selective medium plating technique allowed prediction of the number of cells that would be inactivated by subsequent storage in acidic conditions. SIGNIFICANCE AND IMPACT OF THE STUDY: This work could be useful for improving food preservation by PEF technology and contributes to the knowledge of the mechanism of microbial inactivation by PEF.

Colony Count, Microbial↗

The causes of and cures for contact lens-induced peripheral ulcer.

PURPOSE: Contact lens-induced peripheral ulceration (CLPU) is a relatively common adverse response associated with wearing hydrogel lenses, especially on an extended wear schedule. Bacteriologic examination of lenses at the time of an event has demonstrated an association with Staphylococci spp. We sought to investigate the causes of CLPU in a rabbit model of contact lens wear. METHODS: Rabbits wore contact lenses for a period of 24 hr in the presence or absence of bacteria or in presence or absence of epithelial scratches made in the periphery of the cornea before lens wear. Bacteria tested were a strain of Staphylococcus aureus or a strain of Staphylococcus epidermidis isolated from human CLPUs. Rabbits were also challenged with S. aureus in the presence of an epithelial defect and in the absence of a contact lens. Corneas were monitored by slitlamp, histology, and microbial culture after 24 hr. RESULTS: No CLPU-like lesions were detected under the following conditions: corneal scratch plus lens wear with no bacteria; corneal scratch plus S. epidermidis colonized lenses; corneal scratch without lens wear and with S. aureus applied to eyes; no scratch plus S. aureus colonized lenses; and corneal scratch plus contact lenses colonized by dead S. aureus. CLPU-like lesions were found only when the corneas were scratched and contact lenses colonized by viable S. aureus were applied to the eye. The histology of the lesions demonstrated a frank epithelial break with underlying stromal infiltration. Only low numbers of bacteria could be cultivated from the corneas with CLPU-like lesions. The CLPU-like lesions shared many similarities with CLPU in humans. CONCLUSION: CLPU-like lesions were only produced by S. aureus and not S. epidermidis in the presence of an epithelial abrasion and contact lens. Thus, we hypothesize that to reduce the incidence of CLPU, contact lenses designed to reduce corneal interaction and repel microbial colonization should be produced.

Animals↗

Exploiting molecular methods to explore endodontic infections: Part 1--current molecular technologies for microbiological diagnosis.

Endodontic infections have been traditionally studied by culture-dependent methods. However, as with other areas of clinical microbiology, culture-based investigations are plagued by significant problems, including the probable involvement of viable but uncultivable micro-organisms with disease causation and inaccurate microbial identification. Innumerous molecular technologies have been used for microbiological diagnosis in clinical microbiology, but only recently some of these techniques have been applied in endodontic microbiology research. This paper intended to review the main molecular methods that have been used or have the potential to be used in the study of endodontic infections. Moreover, advantages and limitations of current molecular techniques when compared to conventional methods for microbial identification are also discussed.

Bacterial Typing Techniques↗

A new experimental approach to the search for chemical density factors in the regulation of monoculture growth.

In monocultures of micro-organisms, growth is controlled by feedback mechanisms involving chemical factors such as limiting substrates and inhibitory metabolic products. The role of such feedback in the growth regulation of Escherichia coli O-124 was investigated by growing cells in batch culture using a medium containing glucose and mineral salts. In various phases of growth, portions of the native culture were diluted with culture filtrate, so that although cell density decreased, the chemical composition of the growth medium was unaltered. As the diluted cultures grew, variations in growth acceleration were calculated and compared with those of native (undiluted) cultures. Towards the end of the exponential phase and in the growth deceleration phase, the specific feedback level (FBL) was between -20 and -200 (h g l-1)-1. The feedback components resulting from changes in glucose concentrations were calculated using experimentally determined values of mu max (0.55 +/- 0.05 h-1) and Ks (2.5 +/- 0.7 mg l-1). Only 0.1-40% of FBL could be accounted for by changes in glucose concentration, indicating the presence of additional growth regulators. The method developed may become a new tool for determination of growth-regulating cell-density factors in microbial cultures.

Bacteriological Techniques↗