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Oral microbial flora and oral malodour of the institutionalised elderly in Japan.

OBJECTIVES: To determine the oral health status, especially the level of oral microbial flora and oral malodour, of institutionalised elderly people compared with non-institutionalised elderly people. SETTING: Three institutions and numerous private homes. SUBJECTS: 56 institutionalised and 56 non-institutionalised elderly people. MAIN OUTCOME MEASURES: Institutionalised elderly people had higher levels of candida, staphylococci, and oral malodour. RESULTS: Institutionalised elderly people had a higher level of candida in denture wearers, and a higher level of staphylococci and oral malodour compared with the non-institutionalised elderly. CONCLUSION: The institutionalised elderly have a higher risk of opportunistic infections and oral health care should be improved to protect them from these infections and to reduce oral malodour.

Aged↗

Assessment of chitin decomposer diversity within an upland grassland.

The breakdown of chitin within an acidic upland grassland was studied. The aim was to provide a molecular characterisation of microorganisms involved in chitin degradation in the soil using soil microcosms and buried litter bags containing chitin. The investigation involved an examination of the effects of liming on the microbial communities within the soil and their chitinolytic activity. Microcosm experiments were designed to study the influence of lime and chitin enrichment on the grassland soil bacterial community ex situ under controlled environmental conditions. Bacterial and actinomycete counts were determined and total community DNA was extracted from the microcosms and from chitin bags buried at the experimental site. PCR based on specific 16S rRNA target sequences provided products for DGGE analysis to determine the structure of bacterial and actinomycete communities. Chitinase activity was assessed spectrophotometrically using chitin labelled with remazol brilliant violet. Both liming and chitin amendment increased bacterial and actinomycete viable counts and the chitinase activity. DGGE band patterns confirmed changes in bacterial populations under the influence of both treatments. PCR products amplified from DNA isolated from chitin bags were cloned and sequenced. Only a few matched known species but a prominent coloniser of chitin proved to be Stenotrophomonas maltophilia.

Actinomycetales↗

In vitro antiseptic properties of an ammonium compound combined with denture base acrylic resin.

BACKGROUND: Denture base acrylic resin is easily colonised by oral endogenous bacteria and Candida spp., and eventually by extra-oral species such as Staphylococcus spp., Pseudomonadaceae or members of Enterobacteriaceae. This microbial reservoir can be responsive for denture related stomatitis and aspiration pneumonia, a life-threatening infection especially in geriatric patients. However, the oral and denture hygiene of dependent elderly individuals is extremely poor. OBJECTIVE: This in vitro study aimed to determine the per cent of a quaternary ammonium compound heat-polymerised in acrylic resin necessary to obtain denture base displaying antiseptic properties. DESIGN: Acrylic resin discs containing 2-50% ammonium polymer (Poly 202063A; 0% control) were soaked in artificial saliva for 4 weeks. Resin discs were incubated for 24 hours with Escherichia coli, Staphylococcus aureus, Pseudomonas aeruginosa [37 degrees C, brain-heart infusion (BHI) broth and phosphate-buffered saline (PBS) buffer] and Candida albicans (30 degrees C, Schaedler broth), in 15 ml (168 discs) and 600 microl (168 discs) of inoculum. Microbial growth was verified at t 0 hours and t 24 hours. Data were recorded as the mean of three colony forming unit (CFU) numerations. The borderline of antimicrobial effect was determined at 0.1% viable cells. RESULTS: In 600 microl of PBS inoculum, resin specimens had a bactericidal effect (E. coli and S. aureus: 2%; P. aeruginosa: 10%) and a fungicidal effect (C. albicans: 50%). Long-term stability and toxicity in vivo studies are now required. CONCLUSION: A 2% quaternary ammonium compound polymerised with a denture acrylic resin displayed antiseptic properties after a 4-week soaking period in artificial saliva. Such antiseptic denture base could help geriatric patients to improve their oral health.

Acrylic Resins↗

Molecular analysis of commensal host-microbial relationships in the intestine.

Human beings contain complex societies of indigenous microbes, yet little is known about how resident bacteria shape our physiology. We colonized germ-free mice with Bacteroides thetaiotaomicron, a prominent component of the normal mouse and human intestinal microflora. Global intestinal transcriptional responses to colonization were observed with DNA microarrays, and the cellular origins of selected responses were established by laser-capture microdissection. The results reveal that this commensal bacterium modulates expression of genes involved in several important intestinal functions, including nutrient absorption, mucosal barrier fortification, xenobiotic metabolism, angiogenesis, and postnatal intestinal maturation. These findings provide perspectives about the essential nature of the interactions between resident microorganisms and their hosts.

Animals↗

Usefulness of bacteriological surveillance cultures for monitoring infection in hospitalized patients: a critical reappraisal.

Untargeted bacteriological surveillance of superficial and deep body sites is frequently performed routinely in various clinical settings. This practice is based on the assumption that early identification of surface microbial flora might be predictive of organisms that will later cause invasive disease and that it may consequently assist in guiding empirical antibiotic therapy. A comprehensive review of the literature however indicates that the clinical value and cost-effectiveness of such practices still remain debated and appear largely unproven in most conditions and situations where they are routinely advocated. The present article reviews and critically discusses the available body of evidence supporting or disproving the use of bacteriological surveillance cultures. It is also aimed to issue general recommendations, strategies and methodologies that could be applied in different hospital care settings including the neonatal or adult intensive care as well as the hematology-oncology units.

Belgium↗

Microbial succession of Debaryomyces hansenii strains during the production of Danish surfaced-ripened cheeses.

Surface-ripened cheeses of the Danbo type were analyzed for the presence of yeasts with special emphasis on Debaryomyces hansenii. Samples were taken from pasteurized milk, brine, and inoculation slurries and from cheese surfaces during ripening at a Danish dairy. D. hansenii was found to be the dominant yeast species throughout the ripening period, whereas other yeast species such as Trichosporon spp., Rhodotorula spp., and Candida spp. were found in minor concentrations during early stages of cheese ripening. Mitochondrial DNA RFLP was used to show that several strains of D. hansenii were present from the onset of ripening. Thereafter, a microbial succession among the strains took place during the ripening. After 3 d of ripening, only one strain was found. This particular strain was found to be dominant in 16 additional batches of surface-ripened cheeses. We investigated the cause of the observed microbial succession by determining the variation in strains with regard to their ability to grow on lactate and at different pH and NaCl concentrations. The strains were shown to vary in their ability to grow on lactate. In a full factorial design at three levels with factor levels close to the actual levels on the cheese surface, differences in pH and NaCl tolerances were observed. The dominant strain was found to be better adapted than other strains to the environmental conditions existing in surface-ripened cheeses during production [e.g., lactate as the main carbon source, pH 5.5 to 6.0 and NaCl concentrations of 7 to 10% (wt/vol)].

Adaptation, Physiological↗

Dental caries: an infectious and transmissible disease where have we been and where are we going?

Dental caries is often referred to as an infectious disease, which means different things to different people. Pediatric dentists may be tempted to liken caries to other childhood diseases they encounter yet treat caries from mostly a mechanical or surgical approach. On the other end of the spectrum are clinicians who view the oral cavity as a site teaming with bacteria and think that reducing the overall microbial mass using antimicrobial agents will somehow affect this infectious disease. Here, we argue for another view, that is, to combine the most effective portion of each therapy to yield an effective outcome: the treatment and prevention of dental caries. Because primary teeth are eventually shed, pediatric dentists may be in an advantageous position to implement this approach since restorations to primary teeth need not be as durable as those made to permanent teeth. Moreover, pediatric dentists use sealants extensively and are used to working, for example, with rubber dams, especially in young patients. It may turn out that after more research is completed on the ontogeny of the oral microbial flora, changes or ecological shifts made in childhood become lifelong. More specifically, drastic reductions or eliminations of cariogenic bacteria, along with their sites for colonization in the primary dentition, may prevent or delay their transfer to the permanent dentition. Similarly, reductions in a mother's levels of cariogenic bacteria may delay or interrupt the transfer of cariogenic bacteria to her infant. Here, the goal is to discuss several concepts before their final inception into practices and to stimulate discussion of ideas, reframing the design to meet the demands of everyday practice. Ultimately, the proof will come from a large clinical trial conducted in many practices and patient populations.

Anti-Bacterial Agents↗

Effects of modified atmosphere and vacuum packaging on microbiological and chemical properties of rainbow trout (Oncorynchus mykiss) fillets.

Microbial (psychrotrophic, mesophilic aerobic bacteria and Enterobacteriacae counts), and chemical analysis [pH, total volatile bases nitrogen (TVB-N), lipid oxidation (Thiobarbituric acid reactive substance, TBARS)] of rainbow trout (Oncorynchus mykiss) fillets in air (control), vacuum and modified atmosphere packaging (MAP) with various gas mixtures conditions at 4+/-1 degrees C were determined. The gas mixtures evaluated were 100% CO2, 2.5% O2+7.5% N2+90% CO2 and 30% O2+30% N2+40% CO2. Psychrotrophic bacteria count was above 1 x 10(7) cfu/g on the 12th day in 100% CO2. However; mesophilic bacteria count was below 1 x 10(6) cfu/g at the end of the 14-day storage period. Enterobacteriaceae count was significantly lower in samples packaged with MAP. Lipid oxidation increased rapidly after 6 days of storage in the samples containing 30% O2. While minimum TBARS values were recorded in fillets containing 100% CO2 and vacuumed fillets, the lowest TVB-N values were obtained in fillets with 100% CO2.

Animals↗

Endoscope washers--a protocol for their use.

A protocol for the disinfection of gastroduodenoscopes, retrograde cholangiopancreatography endoscopes and colonoscopes using endoscope washers is described. The process recommends initial manual washing with a disinfectant containing didecyldimethylammonium chloride, surfactants and enzymes, a second washing in the endoscope washer using a detergent associated with a bacteriostatic, fungistatic substance (benzoisothiazolone) and finally the use of a 2% glutaraldehyde product buffered at pH6. After treatment with 2% Steranios added to the washer, less than 1 micro-organism/ml liquid was found in the following units: 83% of the colonoscopes, 83% of the oesophagogastroduodenoscopes, 83% of the main channels of the retrograde cholangiopancreatography endoscopes and 75% of the auxiliary channels of the latter instruments. In 14% of the colonoscopes, 42% of the gastroduodenoscopes, 42% of the main and 50% of auxiliary channels in the retrograde cholangiopancreatography endoscopes there were no signs of microbial growth in the wash liquid. The results obtained indicated that this protocol allowed adequate disinfection of the endoscope channels, structurally the most difficult part of the instrument to disinfect. Emphasis is given to the degree by which instrument contamination can increase during overnight storage, suggesting that endoscopes need to be submitted to further disinfection after overnight storage. Moreover, the water flowing into the washers can also cause instrument recontamination, particularly during the final rinses. Therefore, to better safeguard the health of patients undergoing endoscopy, special care must be taken to maintain the filters and disinfect the washers themselves, no matter how effective the disinfection protocol used may be.

Cholangiopancreatography, Endoscopic Retrograde↗

[An ecologic hygiene assessment of the microbiological processes in soil contaminated with sulfonol and lead].

Alteration of the microbial coenosis and biological activity of different types of soils as well as peculiarities of survival of sanitary-significant and pathogenic microorganisms under the effect of sulphonol and a sulphonol-lead complex have been studied in the laboratory model experiment. The most sensitive tests were as follows: the number of nitrifying bacteria; nitrifying, proteolytic, dehydrogenase and cellulase activity; survival of salmonella and lactose-positive colibacilli. Under the effect of the tested sulphonol (0.5 and 3 mg/l) and lead (0.1 and 2 mg/l) concentrations the self-purification processes in soil were not substantially disturbed, though surfactants negatively affected viability of enterobacteria.

Benzenesulfonates↗

Influence of processing schemes on indicative bacteria and quality of fresh aquacultured catfish fillets.

Fresh aquacultured catfish fillets were obtained from three processors using different processing protocols in summer, autumn, winter, and spring and evaluated for microbial quality. Twenty freshly processed fillets were randomly selected and each fillet was placed in a sterile polyethylene bag. The fillets were transported on ice-pack overnight by air immediately after processing. Five fillets were randomly selected for microbial assays. Each fillet was weighed and an equal volume of sterile 0.1% peptone water at 0 to 1 degrees C was added aseptically. The fillet was massaged (or rinsed) for 120 s and the rinse was used to determine microbial quality. Aerobes (incubation at 35 degrees C for 48 h) and psychrotrophs (incubation at 20 degrees C for 96 h) were enumerated using 3M Petrifilm Aerobic Count plates. Escherichia coli (incubation at 35 degrees C for 24 to 48 h) and total coliforms (incubation at 35 degrees C for 24 to 48 h) were enumerated on 3M Petrifilm E. coli Count plates. Staphylococcus aureus counts were determined on Baird-Parker agar (incubation at 35 degrees C for 48 h). Significant differences (P < or = 0.05) in aerobic, psychrotrophic, total coliform, E. coli, and S. aureus counts due to temperature effects during production and variations in processing protocols were observed. E. coli and S. aureus counts were significantly different during the four seasons. E. coli and S. aureus counts were high during summer and low during winter weather. There was a significant difference (P < or = 0.05) in aerobic, psychrotrophic, and total coliform counts among the three processors during warm weather; however, these differences were significantly (P < or = 0.05) reduced in cold weather.

Animals↗

Basin-wide analysis of the dynamics of fecal contamination and fecal source identification in Tillamook Bay, Oregon.

The objectives of this study were to elucidate spatial and temporal dynamics in source-specific Bacteroidales 16S rRNA genetic marker data across a watershed; to compare these dynamics to fecal indicator counts, general measurements of water quality, and climatic forces; and to identify geographic areas of intense exposure to specific sources of contamination. Samples were collected during a 2-year period in the Tillamook basin in Oregon at 30 sites along five river tributaries and in Tillamook Bay. We performed Bacteroidales PCR assays with general, ruminant-source-specific, and human-source-specific primers to identify fecal sources. We determined the Escherichia coli most probable number, temperature, turbidity, and 5-day precipitation. Climate and water quality data collectively supported a rainfall runoff pattern for microbial source input that mirrored the annual precipitation cycle. Fecal sources were statistically linked more closely to ruminants than to humans; there was a 40% greater probability of detecting a ruminant source marker than a human source marker across the basin. On a sample site basis, the addition of fecal source tracking data provided new information linking elevated fecal indicator bacterial loads to specific point and nonpoint sources of fecal pollution in the basin. Inconsistencies in E. coli and host-specific marker trends suggested that the factors that control the quantity of fecal indicators in the water column are different than the factors that influence the presence of Bacteroidales markers at specific times of the year. This may be important if fecal indicator counts are used as a criterion for source loading potential in receiving waters.

Animals↗

Microflora in oral ecosystems in subjects with hyposalivation due to medicines or of unknown origin.

PURPOSE: To analyse the microbial flora in specific sites in 20 dentate (> or = 16 teeth) subjects with hyposalivation due to medicines or of unknown origin and in 20 controls matched according to age, sex and number of teeth. MATERIALS AND METHODS: The microbial samples were analysed for the following micro-organisms: mutans streptococci, Lactobacillus spp. and Actinomyces spp., associated with the development of caries, Fusobacterium nucleatum and Prevotella intermedia/Prevotella nigrescens, associated with plaque accumulation and gingivitis, Actinobacillus actinomycetemcomitans and Porphyromonas gingivalis, frequently found in elevated numbers in periodontitis sites and Candida albicans, Staphylococcus aureus, enterics and enterococci, associated with mucosal infections. The total microbial count was also registered, together with the total number of streptococci, associated with good oral health. RESULTS: In the hyposalivation group, the mean age was 54 +/- 8 years and the mean number of teeth 25 +/- 3. The unstimulated secretion rate was 0.04 +/- 0.04 mL/min and the stimulated secretion rate 0.98 +/- 0.51 mL/min. Compared with the controls, the hyposalivation group tended to harbour a lower number and proportion of F. nucleatum on the tongue, and an increased number of enterococci in the vestibular region. In the supragingival plaque, the numbers of Strepococcus mutans and Lactobacillus spp. were significantly increased and the number of C. albicans tended to be increased. CONCLUSION: The results indicate that subjects with hyposalivation due to medicines or of unknown origin have a supragingival plaque associated with the development of caries.

Anti-Inflammatory Agents↗

Indicative and pathogenic microbiological quality of aquacultured finfish grown in different production systems.

The nature and number of indicator and pathogenic microbes in fish reared using recirculating and nonrecirculating water systems were compared. For each system, 20 samples of rainbow trout (Oncorhynchus mykiss), tilapia (Oreochromis spp.), hybrid striped bass (Morone saxatilis x M. chrysops), and pacu (Piaractus mesopotamicus) were randomly selected and gutted, and microbial analyses were performed using AOAC procedures. Five fish were subsampled and analyzed for indicative microbial quality with 3M Petrifilm (). The general microbial quality differed significantly (P < 0.05) among the production systems, except for total coliform counts. Rainbow trout cultured in recirculating and nonrecirculating water systems had lower counts for aerobes (2.00 to 3.11 log CFU/g) (p < 0.05), than other species, whereas trout reared in a recirculating water system had significantly lower psychrotrophic numbers (0.86 to 1.85 log CFU/g). Pacu had the highest fecal coliform counts (2.74 to 3.70 log CFU/g), whereas hybrid striped bass and rainbow trout grown in nonrecirculating systems had lower fecal coliform counts (0.00 to 1.39 log CFU/g). Rainbow trout grown in a nonrecirculating system had significantly higher Escherichia coli counts (0.00 to 2.11 log CFU/g). The human bacterial pathogens Listeria monocytogenes, Yersinia enterocolitica, Escherichia coli O157:H7 and Salmonella spp. were not isolated form the fish sampled. However, Clostridium botulinum botulinum was isolated from all the aquacultured fish sampled except pacu and tilapia grown in a recirculating aquaculture system. However, the counts were very low, ranging from 0.0 to 2.3 MPN/g.

Animals↗

Microbial contamination of carcasses and equipment from an Iberian pig slaughterhouse.

The microbial contamination of carcasses and equipment has been studied in an industrial slaughterhouse of Iberian pigs. Samples of the surface of carcasses were taken at different stages of the process and aerobic plate count at 37 degrees C (APC), Enterobacteriaceae-count (E-count) and Escherichia coli-count (EC-count) were determined. It was demonstrated that in scalding and singeing the APC decreased (P < 0.01), while in the dehairing it increased (P < 0.01). The E-count and EC-count decreased in the scalding but increased in the evisceration (P < 0.001). The implementation of good manufacturing practices (GMP) in the stages of closure of the anus and evisceration significantly decreased the EC-count. It changed from 61.1% in carcasses without GMP that had counts higher than 1 log CFU/cm2 to only 7.4% in GMP carcasses. A final wash of the carcasses with potable water at high pressure (the only decontaminating treatment permitted in the European Union) was tested and failed to decrease the counts. It was also demonstrated that cleaning and disinfection of the dehairing and scraping machines is not effective.

Abattoirs↗

Determination of shelf life of concentrated yogurt (labneh) produced by in-bag straining of set yogurt using hazard analysis.

Strained yogurt, labneh, produced by straining cow's milk set yogurt in cloth bags, was stored at 5, 15, and 25 degrees C, and changes in microbial counts, pH, titratable acidity, percentage of free whey, and sensory attributes were monitored during storage. Counts of total aerobes, psychrotrophic yeasts, yeasts and molds, and lactic acid bacteria, except in samples stored at 25 degrees C, increased irrespective of storage temperature. The pH of samples decreased, titratable acidity and percentage of free whey increased, and texture defects were detected at a later stage than flavor changes during storage. Shelf-life data of labneh was adequately described by the Weibull distribution. The nominal shelf life determined using sensory changes and yeast counts as failure criteria ranged from 8.5 to 10.5, 4.7 to 5.8 and 2.3 to 2.7 d at 5, 15 and 25 degrees C, respectively. Q10 (shelf life at T degrees C/shelf life at T+10 degrees C) for flavor quality loss was 1.98 at 5 degrees C, and the corresponding activation energy was 11.3 kcal/mol.

Adult↗

Effect of processing time on enteric bacteria survival and on temperature and chemical composition of broiler poultry litter processed by two methods.

This study was conducted to evaluate the effects of deepstack processing broiler poultry litter with or without aeration on enteric bacteria survival (Trial 1), and to determine the optimum aeration period (Trial 2) and aeration frequency (Trial 3) to control excessive heating of litter during the deepstacking process. The effects of deepstacked litter temperature on bacteria survival and on chemical compositions of the litter were monitored. In Trial 1, E. coli, Salmonella enteriticis and Shigella sonnei were intentionally inoculated into the litter and their survival was monitored with time of litter processing. In Trial 2, litter was deepstacked and aerated once a day from the initial day or from the day when maximum stack temperature was reached and started to decrease to the end of processing period. In Trial 3, litter was deepstacked and aerated once or twice a day from the day after maximum stack temperature was attained until constant temperature was attained. During the deepstacking process of litter pathogenic bacteria were eliminated between the 2nd and 4th day of deepstacking. This phenomenon occurred apparently not only by high heat generated, but also by other potential factors, such as ammonia and microbial competition. Aeration resulted in the heat dissipation and early temperature reduction of deepstacked litter. The optimum aeration procedure to avoid excessive heating of deepstacked litter was to aerate once a day after the stack temperature reached its maximum at the early stage of deepstacking and for 3 days or until the stack temperature dropped off and reached constancy. This aerating frequency and duration did not negatively affect the chemical composition of deepstacked litter. These results suggest that poultry litter properly deepstacked, with or without aeration, to ensure the elimination of enteric bacterial pathogens may be fed at least 10 days earlier than the 21-day deepstacking period previously recommended.

Air↗

Specific and rapid detection by fluorescent in situ hybridization of bacteria in clinical samples obtained from cystic fibrosis patients.

We report on the rapid and specific detection of bacteria commonly isolated from clinical specimens from cystic fibrosis (CF) patients by fluorescent in situ hybridization (FISH). On the basis of comparative sequence analysis, we designed oligonucleotide probes complementary to species-specific 16S rRNA regions of these microorganisms and demonstrated the specificities of the probes by hybridization of different remotely related as well as closely related reference strains. Furthermore, in a pilot project we investigated 75 sputum samples and 10 throat swab specimens from CF patients by FISH and detected Pseudomonas aeruginosa, Burkholderia cepacia, Stenotrophomonas maltophilia, Haemophilus influenzae, and Staphylococcus aureus within these specimens. The specificity of FISH was 100% in comparison to the results of conventional microbial culture. In contrast, the sensitivity of standard laboratory cultivation was moderately higher, since the limit for microscopic detection of bacteria within sputum samples by FISH was approximately 4 x 10(5) CFU/ml of sputum (resulting in a 90% sensitivity for FISH). Moreover, we demonstrated that FISH will be useful for the rapid detection of bacteria that cause acute pulmonary exacerbations in CF patients, as demonstrated in patients with H. influenzae, S. aureus, and P. aeruginosa exacerbations. Therefore, FISH is a valuable additional method for the rapid and specific detection of bacteria in clinical samples from CF patients, in particular, patients with pulmonary exacerbations.

Adult↗