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PubMedDate not supplied

Anaerobic microflora under Class I and Class II composite and amalgam restorations.

OBJECTIVES: The microflora around and beneath restorations is an important factor of restoration failure. The aim of this pilot study was to determine and compare the microbial spectrum under composite and amalgam restorations with special attention to the anaerobic flora. METHOD AND MATERIALS: Ten composite and five amalgam restorations scheduled for replacement were clinically evaluated for marginal gaps, fractures, and secondary caries. After their removal and caries diagnosis, a dentin sample just below the restoration was taken under sterile conditions, stored in a prereduced transport medium for anaerobic bacteria, and immediately transferred to a laboratory for microbial diagnosis. RESULTS: The clinical parameters showing mostly moderate marginal imperfections and the ratios of aerobic to anaerobic flora were comparable for composite and amalgam restorations (11.4%:88.6% and 15.4%:84.5%, respectively). The microbial variety under composite restorations was much greater compared to amalgam, and it was similar to that of infected root canals including anaerobic gram-negative rods, such as Fusobacterium species or Porphyromonas species. Beneath amalgam, the microbial flora was similar to the one found in carious dentin and plaque, with anaerobic and facultatively anaerobic gram-positive rods dominating. Quantitatively, there were up to eight times more microorganisms under composite restorations. The number of bacterial strains correlated with the caries activity and the filling material, the number of anaerobic rods correlated highly with caries activity and localization. In a linear regression, caries activity and the filling material had statistically significant influence on the bacterial load. CONCLUSION: Although caries activity and location had the greatest influence on the microbial flora under the restorations, the kind of restoration material seemed to have an additional effect on the composition of the microflora. This pilot study indicates that inadequate composite restorations may promote the growth of cariogenic, as well as obligate anaerobic and potentially pulpopathogenic bacteria, which should be confirmed by further studies.

Bacteria, Aerobic↗

PubMedDate not supplied

Bacterial adhesion: considerations within a risk-based approach to cleaning validation.

Pharmaceutical manufacturing processes are vulnerable to varying degrees of microbial challenge (hazard) quantifiable as microbial ingress, and microbial retention risks affecting raw materials and inputs to the final product. Control over these risks is exacted by both purposefully designed and incidental (or fortuitous) properties of the manufacturing processes. Within the manufacturing environment, equipment cleaning and hold processes are uniquely prone to microbial challenge yet paradoxically demonstrate the greatest potential for mitigation of these risks. Cognition of those components and contributing factors associated with microbial challenge are necessary to facilitate scientifically sound risk assessments. In the context of equipment cleaning and hold processes, risk assessments are necessary to identify and contrive conditions, which are truly worst case for the validation of the control of microbial challenge. A number of components contribute to the risk of microbial retention, yet the phenomenon of microbial adhesion to surfaces remains one of the most ubiquitous and perplexing. The dual purpose of this review is to primarily précis and provide in a single reference those multi-factorial features and variables contributing to bacterial adhesion, and secondly to provide a guide for interpretation of those considerations for integration into a risk-based approach to cleaning validation.

Bacterial Adhesion↗

PubMedDate not supplied

[Experiences with the testing of the microbial count in drinking water from two laying hen battery sheds].

Samples of drinking water were taken from different sites of two laying hen battery cage barns at 10 days during four months. The result of the bacteriological investigation showed large differences. Up to 5000 CFU/ml were found in the water of the pressure supply vessel. Samples taken from the nipple drinkers even showed higher numbers. It seems that the heterogeneous results are not primarily caused by the methods used. It is assumed that there are still unknown factors present in the drinking water supply system of laying hen battery cage barns which influence the germ content in the drinking water.

Animals↗

PubMed2006

Technological characterization of a bacteriocin-producing Lactobacillus sakei and its use in fermented sausages production.

The aim of this paper was the technological characterization of a Lactobacillus sakei strain, able to produce the bacteriocin sakacin P, that was originally isolated from naturally fermented sausages. Experiments were conducted in situ, using MRS-based medium, and in situ, when the strain was inoculated as starter culture in real sausage fermentation. The results obtained underlined that the strain was able to grow in conditions that are commonly used in the production line, and only lactose and high concentrations of NaCl (5% w/v) reduced the capability for bacteriocin production. When inoculated in sausages, the strain showed a good performance, being able to colonize rapidly the ecosystem. A high number of isolates, capable of producing sakacin P, were already isolated after the third day of fermentation, and persisted throughout the course of the fermentation. The inoculated strain also affected other microbial colonization trends; in fact the total bacterial count and fecal enterococci showed a rapid decrease at the end of the fermentation. Moreover, during sensory evaluation, the final sausage product received high scores for the parameters of tenderness and juiciness, with medium acidity and low rancidity. Lastly, the panelists preferred the sausages produced with the L. sakei characterized in this study when compared to a fermented sausage produced with a commercial starter.

Animals↗

PubMed2006

[Microbiological quality of the air in "small gastronomy point"].

The aim of this work was the estimation of microbial contamination of the air in "small gastronomy point". The study included three places, which have been separated on the ground of their function: 1. area of subsidiaries, 2. area of distribution (sale and serving meal), 3. area of consumption. The total numbers of aerobic mesophilic bacteria, yeasts and moulds were determined by sedimentation method. Taxonomy units of fungal aerosol were also estimated. The samples of air were collected in 16 investigation points in the morning (8-8.30) and in the afternoon (14-14.30). Four series of measurements were carried out and in general 128 of air samples were tested. The results showed that numbers of bacteria, yeasts and moulds were variable and received respectively 30-3397, 0-254 and 0-138 cfu x m(-3). Microbial contamination of air changed depending on area character (the highest average count of bacteria occurred in the air of consumption area and fungi in subsidiaries area), time of a day (contamination of the air increased in the afternoon) and determination date. Only in single samples the numbers of bacteria and fungi were higher than recommended level. Pigmentary bacteria had high participation in total count of bacteria and filamentous fungi were represented mostly by Penicillium sp. and Cladosporium sp.

Air Microbiology↗

PubMed2005

A survey on bacteria inhabiting the sea surface microlayer of coastal ecosystems.

Bacterial populations inhabiting the sea surface microlayer from two contrasted Mediterranean coastal stations (polluted vs. oligotrophic) were examined by culturing and genetic fingerprinting methods and were compared with those of underlying waters (50 cm depth), for a period of two years. More than 30 samples were examined and 487 strains were isolated and screened. Proteobacteria were consistently more abundant in the collection from the pristine environment whereas Gram-positive bacteria (i.e., Actinobacteria and Firmicutes) were more abundant in the polluted site. Cythophaga-Flavobacter-Bacteroides (CFB) ranged from 8% to 16% of total strains. Overall, 22.5% of the strains showed a 16S rRNA gene sequence similarity only at the genus level with previously reported bacterial species and around 10.5% of the strains showed similarities in 16S rRNA sequence below 93% with reported species. The CFB group contained the highest proportion of unknown species, but these also included Alpha- and Gammaproteobacteria. Such low similarity values showed that we were able to culture new marine genera and possibly new families, indicating that the sea-surface layer is a poorly understood microbial environment and may represent a natural source of new microorganisms. Genetic fingerprinting showed, however, no consistent differences between the predominant bacterial assemblages from surface microlayer and underlying waters, suggesting that the presence of a stable and abundant neustonic bacterial community is not a common trait of coastal marine environments.

Bacteria↗

PubMed2005

Reducing Salmonella on cantaloupes and honeydew melons using wash practices applicable to postharvest handling, foodservice, and consumer preparation.

Washing conditions that included a soak or brush scrub were evaluated for removal of Salmonella from the surface of smooth (honeydew) or complex (cantaloupe) melon rinds. Melon rinds were spot-inoculated onto a 2.5 cm2 area of rind (squares) with approximately 6.0 log(10) CFU/square of an avirulent nalidixic acid-resistant strain of Salmonella typhimurium. Melons were washed by immersion in 1500 ml of water or 200 ppm total chlorine and allowed to soak or were scrubbed over the entire melon surface with a sterile vegetable brush for 60 s. Inoculated sites, uninoculated sites ("next to" sites) that were adjacent to inoculated sites, and sites on the side of the melon opposite (remote sites) the inoculated site were excised and pummeled in a stomacher for 2 min prior to plating onto tryptic soy or bismuth sulfite agar supplemented with 50 microg/ml nalidixic acid. S. typhimurium was reduced on the rind of cantaloupe by 1.8 log CFU/melon after soaking for 60 s in 200 ppm total chlorine, which was significantly better than the 0.7 log CFU/melon achieved with soaking in water. For both water and 200 ppm total chlorine, scrubbing with a vegetable brush was shown to be significantly (0.9 log CFU/cantaloupe) more effective than soaking alone. When honeydew melons were soaked or scrubbed in water, reductions of 2.8 log CFU/melon or >4.6 log CFU/melon (four of five samples), respectively, were observed. However, when water treatments were used, the presence of Salmonella-positive "next to" and remote sites indicated that bacteria were spread from inoculated site on the rind to uninoculated sites either through the rinse water (40-70 CFU/ml of Salmonella) or scrub brush (400-500 CFU/brush). Transfer to other sites occurred more often with cantaloupe than honeydew melons. This transfer was eliminated when 200 ppm total chlorine was used. When 200 ppm total chlorine was used, Salmonella could not be detected in the water or on the scrub brush. For optimal microbial removal in food service and home settings, melons should be scrubbed with a clean brush under running water. However, to ensure the benefits of brushing, instructions for cleaning and sanitizing brushes must also be emphasized. For food service settings where concentration and pH can be adequately measured, the use of chlorinated water may provide additional benefit.

Chlorine↗

PubMed2005

Intralaboratory reproducibility of the German test method of prEN ISO 15883-1 for determination of the cleaning efficacy of washer-disinfectors for flexible endoscopes.

In Europe, the evaluation of processing flexible endoscopes in washer-disinfectors (WDs) is performed in accordance with prEN ISO 15883-1, which includes determination of cleaning efficacy. Recent data suggest that large differences are found between cleaning processes when the prEN ISO 15883-1 German test model is applied. Therefore, we analysed 12 separate cleaning processes, and each was assessed by determining the means of three experiments. Transparent test tubes used as test pieces (length 2m, lumen 2mm) were contaminated with a mixture of blood and Enterococcus faecium. The WD used was set to perform the cleaning process alone, which consists of three stages (pre-rinse, cleaning, intermediate rinse). Instead of a cleaning agent, demineralized water was used in all three stages of the cleaning process. The mean bacterial load was 10(11.3)cfu/mL (N=12) in the suspensions of test organism, 10(9.1)cfu/mL (N=12) in the test soils and 10(5.8)cfu/mL (N=12) in the controls (recovery). Mean log(10) reduction factors (RF) for each process, i.e. the difference in microbial loads between the control and the processed tubes, were calculated. The 12 cleaning processes with demineralized water led to a mean RF of 2.2 (range: 1.6-3.2) and excellent visible cleanliness of the test pieces. We found that the prEN ISO 15883-1 German test method leads to reproducible and valid results, and is suitable for determining the cleaning efficacy in WDs.

Colony Count, Microbial↗

PubMed1989

Microbial burdens in disposable and nondisposable ventilator circuits used for 24 and 48 h in intensive care units.

One hospital sought to study the differences in using resterilizable permanent versus disposable ventilator circuits and changing the circuits on a 24-h versus a 48-h basis. Over a period of 13 months 656 condensate samples from 92 permanent and 72 disposable circuits were collected and plated by a loop dilution technique. Two samples were collected from the inspiratory limb (humidifier; tubing or nebulizer), and two were collected from the expiratory limb (tubing and trap) of each circuit. Contamination rates were higher for disposable circuits than for permanent circuits and for 48-h changes than for 24-h changes. Results of chi 2 testing by site indicated there was more contamination on the inspiratory and expiratory limbs each with use of disposable circuits than with the use of permanent circuits. The total results (chi 2 analysis) showed significantly greater microbial growth with the use of disposable circuits (permanent versus disposable, P less than 0.001) and extension of time to 48-h changes (24 h versus 48 h, P less than 0.05). In the experience of this hospital permanent circuits proved more advantageous from the standpoint of contamination risk and cost.

Adolescent↗

PubMed1994

Pollutant effects on the microbial ecosystem.

Genetic diversity of a microbial community will inevitably be affected by environmental stress. However, our understanding of the implications of these effects is limited. Genetic exchange between natural microbial communities appears to be a common phenomenon, mediated by a number of microbial processes (conjugation, transformation, and transduction). These mechanisms of change are presumably adaptations to natural environmental perturbation, e.g., the low levels of antibiotics produced by other organisms. However, anthropogenic influences on the environment may be accelerating genetic change within microbiologic ecosystems, beyond these natural adaptation rates. This article highlights some of the perceived risks to ecosystem health and research questions that need to be addressed.

Colony Count, Microbial↗

PubMed2005

Assessing interactions of multiple agrichemicals by using bacterial assemblages in a wetland mesocosm system.

Agrichemicals may enter wetlands located adjacent to or downstream from agricultural fields. We investigated the individual and interactive effects of three agrichemicals [atrazine, chlorpyrifos, and monosodium acid methanearsonate (MSMA)] and methyl mercury on abundance and heterotrophic potential of wetland heterotrophic bacteria assemblages. We used a factorial experimental design, in which chemicals were introduced in all possible combinations to 66 500-liter mesocosms at the Biological Field Station of the University of Mississippi. Methyl mercury was added to bring the total mercury (HG) concentration to 0.4 mg/Kg wet weight at the sediment surface. Atrazine, chlorpyrifos, and MSMA were added at concentrations of 192, 51, and 219 microg/L, respectively. Over 32 days of exposure, microbial heterotrophic activity was sensitive to only the interactive effect of HG*ATR*CPF in the sediments and only CPF in the water. Total bacterial numbers did not exhibit any significant treatment effects. Therefore, the effects of agrichemicals were reflected on cell-specific bacterial heterotrophic activity rather than bacterial population size.

Agrochemicals↗

PubMed2006

Evaluation of freeze-dried kefir coculture as starter in feta-type cheese production.

The use of freeze-dried kefir coculture as a starter in the production of feta-type cheese was investigated. Maturation of the produced cheese at 4 degrees C was monitored for up to 70 days, and the effects of the starter culture, the salting method, and the ripening process on quality characteristics were studied. The use of kefir coculture as a starter led to increased lactic acid concentrations and decreased pH values in the final product associated with significantly higher conversion rates compared to salted rennet cheese. Determination of bacterial diversity at the end of the ripening process in salted kefir and rennet cheeses by denaturing gradient gel electrophoresis technology, based on both DNA and RNA analyses, suggested a potential species-specific inhibition of members of the genera Staphylococcus and Psychrobacter by kefir coculture. The main active microbial associations in salted kefir cheese appeared to be members of the genera Pseudomonas and Lactococcus, while in salted rennet cheese, Oxalobacteraceae, Janthinobacterium, Psychrobacter, and Pseudomonas species were noted. The effect of the starter culture on the production of aroma-related compounds responsible for cheese flavor was also studied by the solid-phase microextraction-gas chromatography-mass spectrometry technique. Kefir coculture also appeared to extend the shelf life of unsalted cheese. Spoilage of kefir cheese was observed on the 9th and 20th days of preservation at 10 and 5 degrees C, respectively, while spoilage in the corresponding rennet cheese was detected on the 7th and 16th days. Microbial counts during preservation of both types of unsalted cheese increased steadily and reached similar levels, with the exception of staphylococci, which were significantly lower in unsalted kefir cheese. All types of cheese produced with kefir as a starter were approved and accepted by the panel during the preliminary sensory evaluation compared to commercial feta-type cheese.

Animals↗

PubMed2002

A cross sectional study of water quality from dental unit water lines in dental practices in the West of Scotland.

OBJECTIVE: To determine the microbiological quality of water from dental units in a general practice setting and current practice for disinfection of units. DESIGN: A cross-sectional study of the water quality from 40 dental units in 39 general practices and a questionnaire of the disinfection protocols used in those practices. SETTING: NHS practices in primarydental care. SUBJECTS: Thirty-nine general practices from the West of Scotland. METHODS: Water samples were collected on two separate occasions from dental units and analysed for microbiological quality by the total viable count (TVC) method. Water specimens were collected from the triple syringe, high speed outlet, cup filler and surgery tap. Each participating practitioner was asked to complete a questionnaire. Results Microbial contamination was highest from the high speed outlet followed by the triple syringe and cup filler. On average, the TVC counts from the high speed water lines at 37 degrees C and for the high speed lines, triple syringe and cup filler at 22 degrees C were significantly higher than that from the control tap water specimens. The study included units from 11 different manufacturers with ages ranging from under one year to over eight years. The age of the dental unit analysed did not appear to influence the level of microbial contamination. Five of the practices surveyed used disinfectants to clean the dental units but these had no significant effect on the microbiological quality of the water. The majority of dental units (25 out of 40) were never flushed with water between patients. A number of different non-sterile irrigants were used for surgical procedures. CONCLUSION: The microbiological quality of water from dental units in general dental practice is poor compared with that from drinking water sources. Suitable sterile irrigants should be used for surgical procedures in dental practice. Further work is required for pragmatic decontamination regimens of dental unit water lines in a general dental practice setting

Colony Count, Microbial↗

PubMed1991

A guinea pig model of experimental airborne tuberculosis for evaluation of the response to chemotherapy: the effect on bacilli in the initial phase of treatment.

The purpose of this study was to develop and evaluate a guinea pig model of experimental airborne tuberculosis for its ability to assess chemotherapeutic regimens for their efficacy against virulent tubercle bacilli in vivo during the initial phase of treatment. The tissues examined included primary lung lesions and the metastatic foci in lung and spleen which result from the naturally occurring bacillaemia. The treatments examined, INH + RIF, INH + EMB, EMB + RIF, were initiated 4 weeks after infection and were continued for 8 weeks. Although minor differences were observed in the time of onset of a significant bactericidal effect or in the rate of decline in the microbial population, all three treatment combinations resulted in a significant reduction in the number of M. tuberculosis H37Rv recovered from primary lung lesions, primary lesion-free lung lobes and spleen. X-rays taken of excised inflated lung lobes showed a relationship between the degree of calcification of primary lung lesions and the number of surviving bacilli.

Animals↗

PubMed1998

Degradation of pectic compounds during pasteurised vegetable juice spoilage by Chryseomonas luteola: a predictive microbiology approach.

Predictive modelling consists in describing effects of environmental factors on microbial growth parameters. With food spoilage bacteria, this approach must be extended to both growth and food damage characterisation. In order to study the incidence of storage temperature on vegetable damage, using predictive microbiology tools, kinetics of pectic compound degradation were studied. Chryseomonas luteola has been chosen because of its ability to grow on post-harvested vegetables. Experiments were performed at refrigerated temperatures (0-10 degrees C) with low initial bacterial charges (10(1)-10(3.5) cfu/ml). Microbial specific growth rate (mu), stability phase before pectic degradation (Sp) and alteration percentage (Ap) were chosen as reference parameters. Then, sub-optimal temperature effects on these three parameters were estimated using modified Ratkowsky model. Results obtained in synthetic medium were compared with data observed in endive juice to appreciate the alteration of vegetable during post-harvest storage.

Bacteria↗

PubMed2001

Isolation of yeasts and enteric bacteria in root-filled teeth with chronic apical periodontitis.

AIMS: The aim of this study was to determine the occurrence and role of yeasts, enteric gram-negative rods and Enterococcus species in root-filled teeth with chronic apical periodontitis, and the antimicrobial effect of iodine potassium iodide (IKI) irrigation. METHODOLOGY: Forty symptom-free root-filled teeth with chronic apical periodontitis were included in the study. The patients were divided into two groups. In group A the canals were filled with calcium hydroxide for 10-14 days after cleaning and shaping; in group B the canals were irrigated with IKI for 5 min after cleaning and shaping followed by a permanent root filling. Microbiological samples were taken from the canals before and after the chemomechanical preparation and after iodine irrigation (group B). RESULTS: Microbes were isolated from 33 of 40 teeth in the initial sampling. Yeasts were isolated from six teeth, three of them together with E. faecalis. Enteric rods (Escherichia coli, Klebsiella pneumoniae and Proteus mirabilis) were present in three teeth and E. faecalis was isolated from 21 of the 33 culture positive teeth, 11 in pure culture. Growth was detected in 10 teeth of the second samples. Six of the 10 cases were E. faecalis, with five being a pure culture. All third samples (after IKI) except one were negative. The number of microbial cells per sample did not correlate with lesion size. Two flare-ups were recorded, both in teeth with a mixed infection. CONCLUSION: The high prevalence of enteric bacteria and yeasts in root-filled teeth with chronic apical periodontitis was established. IKI improved the antimicrobial effect of the treatment.

Bacterial Infections↗

PubMed1998

Antimicrobial activity of Pelargonium essential oils added to a quiche filling as a model food system.

Eight essential oils obtained by steam distillation from the scented leaves of Pelargonium species and cultivars were added at 250, 500 and 1000 ppm to a quiche filling, inoculated with either Saccharomyces ludwigii or Zygosaccharomyces bailii (at 10(8) cfu g-1), Salmonella enteriditis or Listeria innocua (at 10(9) cfu g-1). The quiche fillings were then kept at 25 degrees C for 24 h and the residual number of micro-organisms determined using the pour plate technique. There was an effective antimicrobial activity by the Pelargonium essential oils at 250 ppm, comparable with that of commercial thyme oil, an excellent antimicrobial agent, against Saccharomyces ludwigii and Zygosaccharomyces bailii, and a lesser inhibition compared with commercial thyme against Salm, enteriditis. There was a greater diversity of activity against L. innocua, which was in some cases more effective than commercial thyme oil. At 500 ppm, there was a greatly increased inhibition of microbial growth using the Pelargonium essential oils, which was comparable with that of commercial thyme, clove, geranium and coriander oils. As there is no evidence for the toxicity of any of these novel Pelargonium oils, and their odour does not make the delicately flavoured quiche filling unpalatable, there is a strong potential for their use in food processing.

Colony Count, Microbial↗

PubMed2004

Pseudomonas aeruginosa corneal binding after 24-hour orthokeratology lens wear.

PURPOSE: To examine the effect of short-term 24-hr orthokeratology lens (OKL) wear on Pseudomonas aeruginosa binding, epithelial surface cell morphology, epithelial sheet thickness, and stromal thickness in a rabbit model. METHODS: Seventeen New Zealand white rabbits were treated according to the Association for Research in Vision and Ophthalmology Statement for the Use of Animals in Ophthalmic and Vision Research. Partial membranectomy was performed on all rabbits 1 week before the experiments. Baseline values for epithelial and stromal thickness and epithelial surface cell size were determined by in vivo confocal microscopy in one randomly chosen eye (n = 6). One week later, rabbits were fitted in the same eye with a hyper oxygen-transmissible OKL. Twenty-four hours later, confocal microscopy was repeated. The second group of rabbits (n = 6) was fitted with an OKL in one randomly chosen eye for 24 hr. P. aeruginosa binding to the corneal epithelium was assessed for the control corneas and those exposed to the test lens. Scanning electron microscopy was performed on a third group of rabbits to assess epithelial surface damage (n = 5). RESULTS: There was a statistically significant difference (P<0.001) in P. aeruginosa binding between the control (1.11 +/- 0.74 x 10(5) colony-forming units per cornea) and the OKL-wearing eyes (2.74 +/- 0.69 x 10(5) colony-forming units per cornea). The central epithelium thinned by 6.5% after lens wear (48.2 +/- 1.9 microm to 45 +/- 1.7 microm, P=0.005); however, central stromal thickness increased by 7.3% (322 +/- 22 microm to 345 +/- 29 microm, P=0.006). Compared with the baseline value, central epithelial cell size increased significantly from 1,253 +/- 140 mm(2) to 1,627 +/- 393 mm(2) (29.4%, P=0.02). Scanning electron microscopy showed increased surface epithelial damage associated with OKL wear. CONCLUSIONS: This prospective, masked, pilot study showed that 24-hr hyper oxygen-transmissible OKL wear induced a statistically significant increase in P. aeruginosa binding to the epithelium of the rabbit cornea, accompanied by central epithelial thinning, stromal thickening, and surface cell damage assessed by scanning electron microscopy. Collectively, the data suggest that despite adequate lens oxygen transmissibility, the mechanical pressure inherent in the OKL design exerted on the corneal surface appears to be associated with increased adherence of P. aeruginosa to surface corneal epithelial cells, which may pose an increased risk for lens-related microbial keratitis, especially in overnight (i.e., closed-eye) wearing conditions. Future studies are needed to determine whether these results are similar in human wear and how P. aeruginosa binding during OKL wear compares with other lens-wearing modalities, such as daily or continuous soft lens wear.

Animals↗