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In vitro evaluation of the antimicrobial activity of five root canal sealers.

The aim of the present study was to analyze the antimicrobial properties of five endodontic sealers: Endo Fill, Endomethasone, Endomethasone N, Sealer 26 and AH-Plus, against the following microorganisms: Candida albicans, Staphylococcus aureus, Enterococcus faecalis, Streptococcus sanguis and Actinomyces naeslundii. The sealers were tested immediately, 24 h, 48 h and 7 days after manipulation.The direct contact method through the observation of the microbial growth in liquid medium and the agar diffusion test were used to evaluate the antimicrobial properties of the sealers. The results, in both methodologies used, showed that immediately after manipulation, Endo-Fill and Endomethasone demonstrated the highest antimicrobial activity, with no statistically significant difference between them. Sealer 26 demonstrated the lowest antimicrobial activity. At all other times after manipulation, there were no statistically significant differences among all the sealers tested. In conclusion, none of the sealers totally inhibited the growth of the microorganisms. Furthermore, the antimicrobial activity of each sealer decreased with time and was dependent upon the microbial susceptibility to them.

Actinomyces↗

Effect of prior serial in vivo passage on the frequency of Salmonella enteritidis contamination in eggs from experimentally infected laying hens.

Experimental infection models are valuable tools for understanding and preventing the deposition of Salmonella enteritidis inside eggs. Oral inoculation is believed to closely simulate naturally occurring S. enteritidis infections of chickens, but oral infection studies have often generated relatively low frequencies of egg contamination. The present study assessed whether repeated in vivo passage of an S. enteritidis strain could affect its ability to cause egg contamination in experimentally infected hens. The incidence of egg contamination was determined in groups of hens inoculated orally with either a phage type 13a S. enteritidis strain or derivatives of this parent strain that were obtained by three successive rounds of passage and reisolation from tissues of infected hens. Passaged S. enteritidis isolates recovered from ovaries and oviducts induced a significantly higher incidence of egg contamination (16.97%) than was attributed to the parent strain (8.27%). However, passaged S. enteritidis isolates recovered from livers and spleens were not associated with a significantly increased frequency of deposition in eggs. By either inducing or selecting for the expression of relevant microbial properties, passage of S. enteritidis through reproductive tissues of chickens may be useful for improving the efficiency at which experimental infection models produce egg contamination.

Administration, Oral↗

Inactivation of Listeria innocua inoculated in liquid whole egg by high hydrostatic pressure.

The resistance of Listeria innocua, as a model microorganism for Listeria monocytogenes, to high hydrostatic pressure in liquid whole egg was studied at several pressures (300, 350, 400, and 450 MPa),temperatures (- 15, 2, and 20 degrees C), and times (5, 10, and 15 min). Listeria innocus was added to liquid whole egg at approximately 10(6) CFU/ml. Listeria innocua was not totally inactivated in any of the treatments. In general, reduction was better at 2 degrees than at room temperature, but the greatest inactivation was obtained at 450 MPa at 20 degrees C for 15 min (over 5 log of reduction), The results indicate that microbial inactivation was increased with prolonged exposure to pressure. D values for Listeria innocua were obtained at 400 MPa for two temperatures (2 and 20 degrees C), and different times (0 to 20 min). The microbial inactivation followed apparent first-order kinetics, exhibiting a decimal reduction time of 7.35 min at 2 degrees C and 8.23 min at 20 degrees C.

Animals↗

Microbiological study of ready-to-eat salad vegetables from retail establishments uncovers a national outbreak of salmonellosis.

The increasing availability of bagged prepared salad vegetables reflects consumer demand for fresh, healthy, convenient, and additive-free foods that are safe and nutritious. During May and June 2001 a study of retail bagged prepared ready-to-eat salad vegetables was undertaken to determine the microbiological quality of these vegetables. Examination of the salad vegetables revealed that the vast majority (3,826 of 3,852 samples; 99.3%) were of satisfactory or acceptable microbiological quality according to Public Health Laboratory Service microbiological guidelines, while 20 (0.5%) samples were of unsatisfactory microbiological quality. Unsatisfactory quality was due to Escherichia coli and Listeria spp. (not Listeria monocytogenes) levels in excess of 10(2) CFU/g. However, six (0.2%) samples were of unacceptable microbiological quality because of the presence of Salmonella (Salmonella Newport PT33 [one sample], Salmonella Umbilo [three samples], and Salmonella Durban [one sample]) or because of a L. monocytogenes level of 660 CFU/g, which indicates a health risk. In each case, the retailer involved and the UK Food Standards Agency were immediately informed, and full investigations were undertaken. Nineteen cases of Salmonella Newport PT33 infection were subsequently identified throughout England and Wales. The outbreak strain of Salmonella Newport PT33 isolated from the salad and from humans had a unique plasmid profile. Campylobacter spp. and E. coli O157 were not detected in any of the samples examined. The presence of Salmonella, as well as high levels of L. monocytogenes, is unacceptable. However, minimally processed cut and packaged salad is exposed to a range of conditions during growth, harvest, preparation, and distribution, and it is possible that these conditions may increase the potential for microbial contamination, highlighting the necessity for the implementation of good hygiene practices from farm to fork to prevent contamination and/or bacterial growth in these salad products.

Colony Count, Microbial↗

Spread of marker bacteria from the hides of cattle in a simulated livestock market and at an abattoir.

The spread of microbial contamination on the hides of beef was investigated at two stages in the meat chain: (i) in a simulated livestock market ("the market") using 33 animals, and (ii) in the unloading-to-skinning area of a commercial abattoir using 18 animals. At both stages, harmless bacterial markers (nalidixic acid-resistant Escherichia coli K-12; rifampicin- and nalidixic acid-resistant Pseudomonas fluorescens; and a tetracycline-resistant E. coli) were inoculated on the hides of a small number of selected animals, and their transfer to other animals and the environment was examined. At the market, the initial prevalence of animals positive for the hide markers (9.1% in each phase) introduced in the presale pen, sale ring, and postsale pen changed to 39.4, 15.1, and 54.5%, respectively, by the end of the market process. In addition, widespread contamination of the market environment with the hide markers was observed. At the abattoir, the initial prevalence of animals positive for the hide marker (11.1%) inoculated at unloading increased to 100% (hide before skinning) and 88.8% (skinned carcass). In addition, another marker inoculated on environmental surfaces in lairage pens, races, and stunning box was detected on 83.3% (hide before skinning) and 88.8% (skinned carcass). These results, although obtained with a relatively small number of animals, demonstrate that both the livestock market process and the unloading-to-skinning process at abattoirs can facilitate the extensive spread of microbial contamination on hides not just within, but also between, batches of animals.

Abattoirs↗

Interrelationships among microbiological, physicochemical, and biochemical properties of Terrincho cheese, with emphasis on biogenic amines.

Changes in the microbiological, physicochemical, and biochemical characteristics of Terrincho cheese as represented by native microflora, pH, water activity, soluble nitrogen fractions, free amino acids, and biogenic amines (e.g., ethylamine, dimethylamine, tryptamine, phenylethylamine, putrescine, cadaverine, histamine, tyramine, cystamine, and spermine) during ripening were monitored. Terrincho is a traditional Portuguese cheese manufactured from raw ewe's milk. The main groups of microorganisms (lactococci, lactobacilli, enterococci, pseudomonads, staphylococci, coliforms, yeasts, and molds) were determined following conventional microbiological procedures. Free amino acids and biogenic amines were determined by reverse-phase high-performance liquid chromatography, following extraction from the cheese matrix and derivatization with dabsyl chloride. The total content of free amino acids ranged from 1,730 mg/kg of dry matter at the beginning of the ripening stage to 5,180 mg/kg of dry matter by day 60 of ripening; such an increase was highly correlated with the increase of water-soluble nitrogen in total nitrogen, 12% trichloroacetic acid-soluble nitrogen in total nitrogen, and 5% phosphotungstic acid-soluble nitrogen in total nitrogen throughout ripening. Histamine was consistently present at very low levels, whereas putrescine, cadaverine, and tryptamine were the dominant biogenic amines and increased in concentration during ripening. Ethylamine, tryptamine, phenylethylamine, and cystamine reached maxima by 30 days of ripening and decreased thereafter. Significant correlations between amino acid precursors and corresponding biogenic amines, as well as between biogenic amines and microbial viable numbers, were observed.

Amino Acids↗

Viruses in Antarctic lakes.

Water samples collected from four perennially ice-covered Antarctic lakes during the austral summer of 1996-1997 contained high densities of extracellular viruses. Many of these viruses were found to be morphologically similar to double-stranded DNA viruses that are known to infect algae and protozoa. These constitute the first observations of viruses in perennially ice-covered polar lakes. The abundance of planktonic viruses and data suggesting substantial production potential (relative to bacteria] secondary and photosynthetic primary production) indicate that viral lysis may be a major factor in the regulation of microbial populations in these extreme environments. Furthermore, we suggest that Antarctic lakes may be a reservoir of previously undescribed viruses that possess novel biological and biochemical characteristics.

Antarctic Regions↗

Bacteriological validation of a new apparatus for disinfection of hospital waste at the point of disposal.

OBJECTIVES: To evaluate the ability of a new apparatus (Dipsys 25, Société SGN, Bagnols sur Cèze, France) to disinfect biomedical waste, including both potentially infectious agents and the normal saprophytic flora of the waste. METHODS: Disinfection was assessed using standard methods (reference strains were fixed on reference carriers according to the French AFNOR methods) and nonstandard assays. Assays in conditions of hospital use, evaluations of bacterial survival during storage, sporicidal effect, and spore survival during storage were performed in parallel. Finally, bactericidal effect in extreme conditions (association of high contamination and high bacterial protection conditions) was tested with normal fecal flora. Bacterial counts were performed after treatment by the apparatus and without treatment (controls). All tests were carried out in triplicate. RESULTS: In all treated carriers, a bacterial population decrease of at least 5 log10 was obtained. Assays performed in hospital-use conditions did not show any bacterial growth. Concerning the evaluation of sporicidal effect and spore revival during conservation, a minimum reduction of 5 log10 was observed in all assays performed, without survival. Finally, concerning assays in extreme conditions, the decrease of bacterial population was between 5 log10 and 10 log10 for vegetative anaerobes of normal fecal flora. CONCLUSION: Under our study conditions, the study apparatus reduced the tested microbial populations by a minimal factor of 5 log10. The main advantage of the apparatus is the opportunity to treat contaminated waste inside hospital wards, at the point of initial collection, without pulverization, by nonspecialized staff.

Bacteria, Aerobic↗

Salivary findings, daily medication and root caries in the old elderly.

Root caries is one of the main dental problems among the dentate old elderly. The aim of our study was to evaluate the occurrence of root caries in 196 dentate elderly (mean age 79.3) in relation to their salivary flow rate, buffering capacity and salivary microbial counts. These subjects participated in the oral health component of the Helsinki Aging Study in 1990-1991. Clinical oral examinations included assessment of the state of root surfaces and salivary flow rates, both paraffin-wax-stimulated and unstimulated. Salivary buffering capacity and the growth of salivary mutans streptococci, lactobacilli and yeasts were determined by means of commercially available kits. Root caries occurred in 52% of men and in 35% of women (p < 0.05). Number of decayed (DRS) and decayed or filled (DFRS) root surfaces correlated (r = 0.16 to 0.26) with salivary microbial counts. No correlation appeared between DRS and subjects' salivary flow rates. Basic estimates on the association between DRS and salivary findings showed that microbial counts only produced significant odds ratios, from 2.0 to 3.5. However, in a log-linear model high salivary mutans streptococci and yeast counts together with male gender were associated with greater occurrence of root caries.

Aged↗

Hygiene aspects of modern poultry chilling.

An evaluation was made of six commercial poultry chilling systems in relation to factors affecting microbial-contamination of carcasses. These systems included water immersion chilling, air chilling and air chilling with evaporative cooling using water sprays. Samples of neck skin and body cavity were taken from carcasses, together with samples from the chilling environment. These were examined for total aerobic mesophilic microbes and counts of presumptive coliform bacteria and Pseudomonas spp. at specific points in the chilling process. Physical measurements included surface and deep-muscle temperatures of carcasses, water temperatures and chlorine concentrations in the immersion system and air speed and temperature during air chilling. The results obtained for water immersion chilling confirmed previous experience that the washing effect reduces microbial contamination of carcasses, although initially the numbers of pseudomonads tended to increase. The air chillers varied in design and mode of operation, but had little overall effect on microbial contamination of the skin. When a completely dry process was used, microbial numbers were reduced approximately ten-fold in the body cavity. However, the use of water sprays tended to increase contamination of the cavity, while relatively heavy spraying using non-chlorinated water, resulted in a substantial increase in the numbers of pseudomonads.

Animals↗

Relationship between extent of burn injury and magnitude of microbial translocation from the intestine.

The gut can be a source of sepsis after thermal injury. In the present study the relationship between the extent of burn injury and magnitude of bacterial translocation was investigated. Mice underwent 0%, 10%, 20%, 30%, or 50% total body surface area full-thickness burn and simultaneous gavage with 1 x 10(10) 14C-labeled Escherichia coli. mesenteric lymph nodes, liver, spleen, peritoneal fluid, and burn wound were excised 4 hours after burn injury. Residual radioactivity and bacterial colony counts were measured, and percentages of viable organisms were calculated. Results showed that the rate of translocation of 14C E. coli increased proportionally with the burn size, reaching a maximum at 30%. The cutaneous eschar collected a remarkable amount of labeled bacteria, suggesting enteric microflora as a possible source of contamination of the burn wound via endogenous routes. The percentage of viable organisms in the tissues demonstrated that the ability of mesenteric lymph nodes, liver, and eschar to clear translocated bacteria was directly affected by the severity of the burn injury.

Animals↗

Skin debris and micro-organisms on the periwound skin of pressure ulcers and the influence of periwound cleansing on microbial flora.

Many clinicians use the same solution, most often normal saline, to cleanse the periwound skin and the wound bed itself. However, skin debris such as water-insoluble proteins and lipids are not efficiently removed by normal saline solutions. To analyze the skin debris and micro-organisms found on the periwound skin of pressure ulcers and to evaluate the effect of periwound cleansing on the microbial flora, a descriptive study was conducted among 17 long-term care residents with Stage III and IV pressure ulcers. Skin debris from both the periwound area and normal skin was collected from all 17 residents. In addition, micro-organisms from the wound bed, periwound, and normal skin of five residents were collected before, immediately after, and 6 hours and 24 hours after periwound cleansing using a skin cleanser. All microbial species were identified by cultivation. Cholesterol and nitrogen-containing substances were found in greater quantity on the periwound than on normal skin (P = 0.0027 and P = 0.0054, respectively) and the number of isolated micro-organisms from the periwound area was larger than that from normal skin. Protein showed the highest correlation to the microbial count present on the periwound (r = 0.71, P = 0.0014). The microbial counts of all isolated micro-organisms decreased immediately after cleansing but the number of isolates with high microbial counts increased over time. In the wound bed, the number of isolates with decreasing microbial counts was larger than the number of isolates with increasing microbial counts. Both numbers returned to pre-cleansing values after 24 hours, suggesting that periwound cleansing only (without directly cleansing the wound bed) is effective at reducing the microbial counts in the wound bed for up to 24 hours. Further research is needed to evaluate the effects of periwound cleansing on healing time.

Administration, Cutaneous↗

Microbiological quality of rabbit meat.

World rabbit meat production is estimated to be over 1 million tons, and Spain is the third largest producer. Although rabbit meat is marketed and consumed worldwide, information on microbiological quality is very scarce. Here, we report indicator organisms, spoilage flora, sensory quality, and some physicochemical traits of 24 h postmortem chilled rabbit carcasses and prepackaged rabbit meat stored chilled in air for 0 to 3 days at the retail level. The mean total bacterial count (4.01 +/- 0.48 log CFU/g) for carcasses dressed at a small abattoir by a manual process was significantly lower (P < 0.05) than that (4.96 +/- 0.90 log CFU/g) for carcasses dressed at a large abattoir in automated slaughter lines. Both groups of carcasses had mean pH values of 5.98. The dominant contaminants on carcasses from the small abattoir were Pseudomonas, lactic acid bacteria, and yeasts. These microorganisms and Brochothrix thermosphacta were dominant on carcasses from the large abattoir. On prepacked hind legs (pH 6.26 +/- 0.18) stored at -1 to +1 degree C (supermarket 1), mean aerobic mesophilic count was 5.87 +/- 1.03 log CFU/g, and the major microbial groups were Pseudomonas, yeasts, lactic acid bacteria, and B. thermosphacta. On prepacked whole carcasses (pH 6.37 +/- 0.18) displayed at -1 to +5 degrees C (supermarket 2), mean aerobic mesophilic count was 6.60 +/- 1.18 and the same microbial groups were dominant. Relative Escherichia coli incidence was supermarket 2 > large abattoir > supermarket 1 > small abattoir. Overall, low numbers of coliforms, Enterobacteriaceae, psychrotrophic clostridia, coagulase-positive staphylococci, and molds were found. Sensory scores, pH values, and L-lactic acid content differentiated fresh carcasses from retail samples. Data obtained suggest that the microflora of chilled rabbit meat are different from those found on the meat of other animals.

Abattoirs↗

Response of Bdellovibrio and like organisms (BALOs) to the migration of naturally occurring bacteria to chemoattractants.

A dual culture-based and non-culture-based approach was applied to characterize predator bacterial groups in surface water samples collected from Apalachicola Bay, Florida. Chemotaxis drop assays were performed on concentrated samples in an effort to isolate predator bacteria by their chemotactic ability. Yeast extract (YE) and casamino acids (CA) proved to be strong chemoattractants and resulted in three visibly distinct bands; however, dextrose, succinate, pyruvate, and concentrated cells of Vibrio parahaemolyticus P5 as prey did not elicit any response. The three distinct bands from YE and CA were separately collected to identify the chemotactic microbial assemblages. Plaque-forming unit assays from different chemotaxis bands with P5 as prey indicated 5- (CA) to 10-fold (YE) higher numbers of predator bacteria in the outermost chemotactic bands. Polymerase chain reaction-restriction fragment length polymorphism and 16S rDNA sequencing of clones from different chemotaxis bands resulted in identification of Pseudoalteromonas spp., Marinomonas spp., and Vibrio spp., with their numbers inversely proportional to the numbers of predators-i.e., Bdellovibrio spp. and Bacteriovorax spp-in the chemotaxis bands. This study indicates that predatorial bacteria potentially respond to high densities of microbial biomass in aquatic ecosystems and that chemotaxis drop assay may be an alternate culture-independent method to characterize predatorial bacterial guilds from the environment.

Bacteria↗

Chlorhexidine prophylaxis for chemotherapy- and radiotherapy-induced stomatitis: a randomized double-blind trial.

Patients receiving cytotoxic antineoplastic therapy often have treatment-associated stomatitis. A 0.12% chlorhexidine digluconate mouthrinse was evaluated (15 ml, three times a day) in a prospective, double-blind randomized trial as prophylaxis against cytotoxic therapy-induced damage to oral soft tissues. Seventy subjects, forty inpatients receiving high-dose chemotherapy and thirty outpatients receiving high-dose head and neck radiation therapy, were evaluated. Chlorhexidine mouthrinse significantly reduced the incidence of oral mucositis in the chemotherapy group on day 14 (p less than 0.02) and at 1 week follow-up on day 28 (p less than 0.002). Mucositis in the patients undergoing chemotherapy who received chlorhexidine also resolved more rapidly. Mucositis severity was significantly less compared to the control chemotherapy group on day 14 (p less than 0.03), day 21 (p less than 0.04), and on 1 week follow-up (p less than 0.02). Concomitant trends in the reduction in oral streptococci and yeast were noted in the chemotherapy group receiving chlorhexidine mouthrinse. Although no differences were observed in oral mucositis between the control and chlorhexidine groups of patients undergoing high-dose radiotherapy, similar reductions of oral microflora to those seen in the chemotherapy population were also noted for patients undergoing radiation therapy who received chlorhexidine. Although generally not significant, some increase in gram-negative bacilli was noted in the chlorhexidine-treated patients in both the chemotherapy and radiotherapy groups, but there was no correlation with increased systemic infection. Prophylactic chlorhexidine mouthrinse reduces oral mucositis and microbial burden in patients with cancer undergoing intensive chemotherapy.

Adult↗

Campylobacter jejuni loss of culturability in aqueous microcosms and ability to resuscitate in a mouse model.

Water is known as one of the main transmission routes of Campylobacter and contributes to increase the number of sporadic infections and outbreaks. Campylobacter jejuni persists in the environment, especially in water, in a viable but non-culturable (VBNC) form that is thought to be a possible cause of water-borne outbreaks. In this study, we evaluated the loss of culturability and viability of 9 C. jejuni strains of clinical origin and one ATCC reference strain when kept at 4 degrees C in artificial sea water (ASW). Culturability was measured as colony-forming units while viability was evaluated by CTC-DAPI double staining and the combined CTC-specific fluorescent antibody technique (CTC-FA). When cultured on Columbia Agar plates, strains exhibited different growth profiles which allowed to classify them into three different groups. Both techniques used to monitor the viability of the bacterial cells showed that C. jejuni strains survived in the VBNC form in the microcosms through a period lasting from 138 to 152 days. The recovery of C. jejuni VBNC forms to culturability, as evidenced by cell division, was obtained by passage in the mouse intestine. Our results indicate that C. jejuni VBNC cells were able to remain in this state for a few months and regain their culturability after in vivo passage depending on their lasting in the VBNC state, which affects the number of respiring bacteria. In fact, the resuscitation was achieved when the number of respiring bacteria became higher than 10(4) cell/ml. Therefore, a relatively high microbial titer of respiring bacteria in the VBNC state seems to be important for the resuscitation and subsequent intestinal colonisation.

Animals↗

Effect of acid adaptation on inactivation of Salmonella during drying and storage of beef jerky treated with marinades.

This study evaluated the influence of pre-drying marinade treatments on inactivation of acid-adapted or nonadapted Salmonella on beef jerky during preparation, drying and storage. The inoculated (five-strain composite, 6.0 log CFU/cm2) slices were subjected to the following marinades (24 h, 4 degrees C) prior to drying at 60 degrees C for 10 h and aerobic storage at 25 degrees C for 60 days: (1) no marinade, control (C), (2) traditional marinade (TM), (3) double amount of TM modified with added 1.2% sodium lactate, 9% acetic acid, and 68% soy sauce with 5% ethanol (MM), (4) dipping into 5% acetic acid and then TM (AATM), and (5) dipping into 1% Tween 20 and then into 5% acetic acid, followed by TM (TWTM). Bacterial survivors were determined on tryptic soy agar with 0.1% pyruvate and xylose-lysine-tergitol 4 (XLT4) agar. Results indicated that drying reduced bacterial populations in the order of pre-drying treatments TWTM (4.8-6.0 log CFU/cm2)> or =AATM> or =MM>TM> or =C (2.6-5.0 log CFU/cm2). Nonadapted Salmonella were significantly (P<0.05) more resistant to inactivation during drying than acid-adapted Salmonella in all treatments. Bacterial populations decreased below the detection limit (-0.4 log CFU/cm2) as early as 7 h during drying or remained detectable even after 60 days of storage, depending on acid adaptation, pre-drying treatment, and agar media. The results indicated that acid adaptation may not cause increased resistance of Salmonella to the microbial hurdles involved in jerky processing and that use of modified marinades in manufacturing jerky may improve the effectiveness of drying in inactivating Salmonella.

Adaptation, Physiological↗

Variations in microbial indicator densities in beach waters and health-related assessment of bathing water quality.

Daily and hourly variations in microbial indicators densities in the beach-waters of Hong Kong have been described. The levels of Escherichia coli at a number of beaches was observed to be influenced by tide, and for staphylococci, by bather numbers. The tidal influence was most obvious during spring tides; and for the effect of bathers, during neap tides. Both organisms are present in high densities in external sources of faecal pollution of bathing beaches, with the average staphylococci to E. coli ratios being 0.04-3. Staphylococci may serve as an indicator of bather density and the risk of cross-infection amongst bathers (rather than as another indicator of faecal contamination) when the average staphylococci to E. coli ratio for a bathing beach is considerably higher than 3. The variability of microbial indicator densities means the routine sampling of bathing beaches should be carried out on weekend days with maximum numbers of swimmers exposed to the water, and spread throughout the bathing season.

Colony Count, Microbial↗