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Microbiological evaluation of a newly designed dental air-turbine handpiece for anti-cross contaminations.

The effectiveness of a newly developed anti-cross contamination device for a dental air-turbine handpiece was tested. The handpiece with or without the anti-cross contamination device was contaminated with two bacterial strains, Staphylococcus aureus and Streptococcus mutans, as well as two bacteriophage strains, T2 and MS2. After contamination with these microorganisms, the handpieces were disinfected with glutaraldehyde or replaced with newly autoclaved ones. Residual microorganisms inside the handpiece or an air/water supply hose line were collected and counted after overnight cultivation. The anti-cross contamination device effectively reduced the contamination level of an air-turbine handpiece to that of the negative control. No microbial contamination in the air/water supply hose line was detected with this device.

Air Microbiology↗

Microbial biofilm formation and contamination of dental-unit water systems in general dental practice.

Dental-unit water systems (DUWS) harbor bacterial biofilms, which may serve as a haven for pathogens. The aim of this study was to investigate the microbial load of water from DUWS in general dental practices and the biofouling of DUWS tubing. Water and tube samples were taken from 55 dental surgeries in southwestern England. Contamination was determined by viable counts on environmentally selective, clinically selective, and pathogen-selective media, and biofouling was determined by using microscopic and image analysis techniques. Microbial loading ranged from 500 to 10(5) CFU. ml(-1); in 95% of DUWS water samples, it exceeded European Union drinking water guidelines and in 83% it exceeded American Dental Association DUWS standards. Among visible bacteria, 68% were viable by BacLight staining, but only 5% of this "viable by BacLight" fraction produced colonies on agar plates. Legionella pneumophila, Mycobacterium spp., Candida spp., and Pseudomonas spp. were detected in one, five, two, and nine different surgeries, respectively. Presumptive oral streptococci and Fusobacterium spp. were detected in four and one surgeries, respectively, suggesting back siphonage and failure of antiretraction devices. Hepatitis B virus was never detected. Decontamination strategies (5 of 55 surgeries) significantly reduced biofilm coverage but significantly increased microbial numbers in the water phase (in both cases, P < 0.05). Microbial loads were not significantly different in DUWS fed with soft, hard, deionized, or distilled water or in different DUWS (main, tank, or bottle fed). Microbiologically, no DUWS can be considered "cleaner" than others. DUWS deliver water to patients with microbial levels exceeding those considered safe for drinking water.

Bacteria↗

Bactericidal effects of negative air ions on airborne and surface Salmonella enteritidis from an artificially generated aerosol.

The bactericidal effect of high levels of negative ions was studied using a custom-built electrostatic space charge device. To investigate whether the ion-enriched air exerted a bactericidal effect, an aerosol containing Salmonella Enteritidis (SE) was pumped into a sealed plastic chamber. Plates of XLT4 agar were attached to the walls, top, and bottom of the chamber and exposed to the aerosol for 3 h with and without the ionizer treatment. The plates were then removed from the chamber, incubated at 37 degrees C for 24 h, and colonies were counted. An average of greater than 10(3) CFU/plate were observed on plates exposed to the aerosol without the ionizer treatment (control) compared with an average of less than 53 CFU/plate on the ionizer-treated plates. In another series of experiments, the SE aerosol was pumped for 3 h into an empty chamber containing only the ionizer and allowed to collect on the internal surfaces. The inside surfaces of the chamber were then rinsed with 100 ml phosphate-buffered saline that was then plated onto XLT4 plates. While the rinse from the control chamber contained colony counts greater than 400 CFU/ml of wash, no colonies were found in the rinse from the ionizer-treatment chamber. These results indicate that high levels of negative air ions can have a significant impact on the airborne microbial load, and that most of this effect is through direct killing of the organisms. This technology, which also causes significant reduction in airborne dust, has already been successfully applied for poultry hatching cabinets and caged layer rooms. Other potential applications include any enclosed space such as food processing areas, medical institutions, the workplace, and the home, where reduction of airborne and surface pathogens is desired.

Aerosols↗

Antibacterial effect of tetracaine hydrochloride (Pontocaine) in the aspect of exposure time--in vitro study.

Antibacterial effect of tetracaine hydrochloride was studied. Tetracaine hydrochloride (preservative free) were incubated with Staphylococcus aureus, coagulase negative staphylococcus and Pseudomonas aeruginosa respectively, for 18 hours and for 2 minutes. Then it was diluted and cultured on nutrient agar plate. Colony counts were done after 18 hours. In case of 18 hours incubation, there was no growth of microbials in 0.5%, 0.1% tetracaine hydrochloride, but there was no inhibitory effect of 0.02% tetracaine hydrochloride on growth of microbials, irrespective of inoculum amount. In case of 2 minutes incubation with 0.5% tetracaine hydrochloride, there was no difference between the amount of microbial inoculum and colony count. Above indicates that tetracaine hydrochloride has no inhibitory effect on bacterial growth after short exposure of less than 2 minutes.

Anti-Bacterial Agents↗

Effect of feeding frequency on bacterial and fungal concentrations, pH, and other parameters in the rumen.

Three sheep were fed a pelleted high-roughage diet either once, 6, or 24 times per day in a 3 x 3 Latin square trial. During each 21-d period, 14 d were allowed for adaptation followed by a 7-d collection period, in which samples for microbial counts were taken on d 1 and 5 and several rumen parameters were measured on d 2 and 6. Bacterial concentrations were not different between feeding frequencies on the first sampling day but were higher (P < 0.05) on the second sampling day when the sheep were fed 24 times a day. Fungal concentrations were not different among feeding frequencies on either sampling day. No effects of feeding frequency were observed for the concentration of cellulolytic bacteria or fungi. On d 2, ruminal volume was larger (P < 0.05) with six feedings than with one feeding and fluid volume turnover was greater (P < 0.05) when sheep were fed 24 times per day. Rumen pH values were higher (P < 0.01) on both d 1 and 5 when the sheep were fed once a day and the percentage of rumen dry matter was highest (P < 0.02) with 24 feedings. These findings would suggest that if the same amount of a given diet is fed daily, the number of feedings does not markedly affect microbial concentrations, rumen volume, or liquid turnover time.

Adaptation, Physiological↗

Microbiological quality assurance of purified water by ozonization of storage and distribution system.

Purified water storage and distribution systems at ambient temperature are highly susceptible to microbial contamination and formation of biofilm. The impact of two disinfection regimens with ozone as a function of time, the heterotrophic plate counts (HPC), and the concentration of total organic compounds (TOC) in purified water were investigated over a period of 4 years. We have established that concentrations of ozone of 70 +/- 20 ppb in the production regimen and 250 +/- 50 ppb in the disinfection regimen are sufficient to maintain a low bioburden and low TOC in a recirculating distribution system. The purified water that entered into the distribution system has low HPC (0.01 CFU/mL), indicating a reduction by ozone in the storage tank by up to approximately 120-fold. Over 4 years, 94-98% of the microbial counts were in the category 0-5 CFU/mL, and none in category > or =50 CFU/mL. In spite of increased TOC in the inlet water, up to 40 ppb, the microbial counts in purified water in the distribution loop were unaffected. The study emphasizes that the critical points regarding microbial contamination of the purified water system are user point valves and the tubes used for transferring water to equipment. The specified ozone level prevents microbial growth and formation of biofilm in the distribution system that might otherwise endanger the water quality by sporadic release of microbes.

Bacteria↗

Density and composition of microorganisms during long-term (418 day) growth of potato using biologically reclaimed nutrients from inedible plant biomass.

A study evaluating alternative methods for long term operation of biomass production systems was recently completed at the Kennedy Space Center (KSC). The 418-day study evaluated repeated batch versus mixed-aged production of potato grown on either standard 1/2-strength Hoagland's nutrient solution or solutions including nutrients recycled from inedible plant material. The long term effects of closure and recycling on microbial dynamics were evaluated by monitoring the microbial communities associated with various habitats within the plant growth system (i.e., plant roots, nutrient solution, biofilms within the hydroponic systems, atmosphere, and atmospheric condensate). Plate count methods were used to enumerate and characterize microorganisms. Microscopic staining methods were used to estunate total cell densities. The primary finding was that the density and composition of microbial communities associated with controlled environmental plant growth systems are stable during long term operation. Continuous production resulted in slightly greater stability. Nutrient recycling, despite the addition of soluble organic material from the waste processing system, did not significantly increase microbial density in any of the habitats.

Air Microbiology↗

Surface contamination in the dental operatory: a comparison over two decades.

BACKGROUND: This study compares surface bacterial contamination levels in a large teaching clinic in 1998 with that of a 1976 study in the same clinic to determine if renovation and more stringent infection control, or IC, practices have made a difference. METHODS: The authors obtained samples from surfaces in clinic operatories--including the light handle covers, jacket cuffs, sinks and floors--in the morning and afternoon. They compared the bacterial counts from these samples with those from the 1976 study. Data were analyzed using the matched pairs t tests and independent t tests. RESULTS: In both the 1976 and 1998 studies, mean bacterial counts were higher at the end of the day than in the morning; however, the differences were only significant in the 1976 study. Comparisons between the 1998 and 1976 counts revealed significant differences for the floor, headrest, light handle covers and clinic jacket cuffs. CONCLUSIONS: Improvements in clinic design and equipment, as well as IC procedures and practices resulted in a lower level of surface bacterial contamination in 1998 than in 1976 in a large teaching clinic. Clinical Implications. Attention to clinic design and furnishings and adherence to recommended IC practices can reduce microbial surface contamination, thereby reducing the risk of causing cross-infection and making dental practice safer.

Colony Count, Microbial↗

Rapid specific detection and quantification of bacteria and archaea involved in mineral sulfide bioleaching using real-time PCR.

A SybrGreen real-time PCR assay was developed to detect and quantify both total and selected 16S rDNA species of bacteria and archaea involved in the bioleaching of metals from sulfide ores. A set of specific and universal primers based on 16S rDNA sequences was designed and validated for specific detection and quantification of DNA isolated from representative strains of Acidianus brierleyi, Sulfolobus sp., Sulfobacillus thermosulfidooxidans, Sulfobacillus acidophilus, Acidithiobacillus caldus, and Leptospirillum ferrooxidans. An artificial sequence based on 16S rDNA was constructed to quantify total 16S rDNA in mixed DNA samples. The real-time PCR assay was further validated using a mixture of 16S rDNA amplicons derived from the six different species, each added at a known amount. Finally, the real-time PCR assay was used to monitor the change of 16S rDNA copies of four bioleaching strains inoculated into chalcopyrite airlift column reactors operated at different temperatures. The growth dynamics of these strains correlated well with the expected effects of temperature in the chalcopyrite-leaching environment. The suitability of this method for monitoring microbial populations in industrial bioleaching environments is discussed.

Archaea↗

Competitive and interactions affecting a fermentative spirochete in anaerobic chemostats.

Terminal restriction fragment length polymorphism and fluorescent in situ hybridization revealed that spirochete-related populations dominated two glucose-fed methanogenic bioreactor communities at dilution rates of 0.06, 0.13, and 0.17 day(-1). At dilution rates of 0.25 and 0.50 day(-1), spirochete-related populations decreased while Clostridium-related populations increased. Isolates representing both dominant populations were obtained (Treponema R8 and Clostridium S9) and competed against each other in continuous culture. Treponema R8 out-competed Clostridium S9 at all dilution rates applied (0.17 to 1.0 day(-1)) when sufficient pantothenate was supplied in the medium. Without sufficient pantothenate, the population size of Treponema R8 was limited to 40% of the total cells. Coculture of Treponema R8 with Methanobacterium bryantii increased the cell yield of Treponema R8 and relieved the pantothenate requirement. Triculture of Treponema R8, Clostridium S9, and M. bryantii in pantothenate-deficient medium allowed Treponema R8 to outcompete Clostridium S9 in continuous culture upto a dilution rate of 0.50 day(-1). These experiments demonstrate that cofactor and vitamin requirements can affect the competitive success of a microbial species.

Base Sequence↗

Microbial monitoring of spacecraft and associated environments.

Rapid microbial monitoring technologies are invaluable in assessing contamination of spacecraft and associated environments. Universal and widespread elements of microbial structure and chemistry are logical targets for assessing microbial burden. Several biomarkers such as ATP, LPS, and DNA (ribosomal or spore-specific), were targeted to quantify either total bioburden or specific types of microbial contamination. The findings of these assays were compared with conventional, culture-dependent methods. This review evaluates the applicability and efficacy of some of these methods in monitoring the microbial burden of spacecraft and associated environments. Samples were collected from the surfaces of spacecraft, from surfaces of assembly facilities, and from drinking water reservoirs aboard the International Space Station (ISS). Culture-dependent techniques found species of Bacillus to be dominant on these surfaces. In contrast, rapid, culture-independent techniques revealed the presence of many Gram-positive and Gram-negative microorganisms, as well as actinomycetes and fungi. These included both cultivable and noncultivable microbes, findings further confirmed by DNA-based microbial detection techniques. Although the ISS drinking water was devoid of cultivable microbes, molecular-based techniques retrieved DNA sequences of numerous opportunistic pathogens. Each of the methods tested in this study has its advantages, and by coupling two or more of these techniques even more reliable information as to microbial burden is rapidly obtained.

Adenosine Triphosphate↗

Effect of the tropical grass Brachiaria brizantha (Hochst. ex A. Rich.) Stapf on microbial population and activity in petroleum-contaminated soil.

The effect of the tropical pasture grass Brachiaria brizantha on numbers of bacteria, fungi and degraders of alkanes, aromatics, cycloalkanes and crude oil in petroleum hydrocarbon contaminated and uncontaminated savannah soil was evaluated. Substrate induced soil respiration and soil pH were compared between planted and unplanted soil. B. brizantha had a mostly increasing effect on microbial numbers. As an exception, growth of bacteria was not or negatively affected. Microbial respiration and pH were always lower in planted than in unplanted soil. Low pH may result from enhanced oil degradation in planted soil leading to an accumulation of organic acids. A comparable stimulation of crude oil degraders and fungi in planted soil points to the importance of fungi. Since they tolerate lower pH values than bacteria, they are considered to play a central role in oil degradation. Given that the enhancement of crude oil degradation under the influence of B. brizantha could not clearly be correlated to microbial numbers and activity, other factors like oxygen availability, plant enzymes and synergistic degradation by microbial consortia have to be considered.

Bacteria↗

In vitro assessment of the effectiveness of chlorhexidine gel and calcium hydroxide paste with chlorhexidine against Enterococcus faecalis and Candida albicans.

OBJECTIVE: The purpose of this study was to compare the effectiveness of various medicaments, including Ca(OH)2/2% chlorhexidine, 2% chlorhexidine gel, and Ca(OH)2 alone, against Enterococcus faecalis and Candid albicans in vitro. STUDY DESIGN: Eighty extracted single-rooted human maxillary teeth were used. After removing the crown, each root was instrumented up to size 50 by using a conventional technique. The root canal was irrigated with ethylenediamine tetra-acetic acid (EDTA) solution to remove smear layer. Then, roots were infected with E faecalis and C albicans. Subsequently, the roots were divided into 4 treatment groups: group 1 was treated with calcium powder hydroxide in distilled water, group 2 was treated with calcium hydroxide powder in 2% chlorhexidine, group 3 was treated with 2% chlorhexidine gel, and group 4 was treated with 0.9% sterile saline serving as negative control. Microbial samples were taken after 7, 15, and 30 days. After incubation, dentine chips were obtained form each root canal and examined microbiologically. The microbiological samples were plated to count colony-forming units in per milligram of dentin. RESULTS: The results showed that the 2% chlorhexidine gel was significantly more effective than calcium hydroxide with 2% chlorhexidine, calcium hydroxide, and control saline solution (P < .05). CONCLUSION: Under the conditions of this study, 2% chlorhexidine gel is effective in the elimination of E faecalis and C albicans from the root canal system. However, to support this in vitro observation, further in vivo studies are needed.

Anti-Infective Agents, Local↗

Bacterial recolonization of the skin under a polyurethane drape in hip surgery.

This study investigated the proliferation of skin flora under polyurethane (PU) wound dressings following surgical interventions. In vitro the film proved to be impermeable to staphylococci. However, bacteria migrated under the film, showing a preference for the adhesive rather than the adhesive-free areas. The skin of 43 patients with incisions of the proximal thigh was covered postoperatively with a transparent PU-dressing. This was examined bacteriologically preoperatively and at the third and seventh postoperative days. The use of an antiseptic (povidone iodine) reduced the number of colony forming units in the postoperative skin but at the same time there was a significant reduction of bacteria under the film in comparison with the uncovered skin. The numbers and species of bacteria in different areas under the film did not suggest a spread of organisms from the uncovered skin to the skin under the film, but rather that there was a multifocal microbial regeneration.

Adult↗

Initial effect of controlled release chlorhexidine on subgingival microorganisms.

BACKGROUND: Little or no data exist on the ability of subgingival application of PerioChip (2.5 mg chlorhexidine gluconate in a biodegradable chip; Astra Pharmaceuticals, Westborough, MA, USA) to suppress periodontopathic microorganisms. The present study compared the subgingival microbiota of periodontitis sites receiving the chlorhexidine chip plus scaling and root planing (Sc/Rp) or Sc/Rp alone. METHODS: Seven males and six females, mean age 49 years, with moderate to advanced periodontitis participated in the study. In each patient, two bilateral pockets probing 6-7 mm were randomly assigned to treatment by chlorhexidine chip + Sc/Rp, or by Sc/Rp alone. Subgingival placement of chlorhexidine chips was carried out according to the manufacturer's instructions. Sc/Rp was performed with hand instruments for at least 10 min in each study tooth. Subgingival samples were collected by paper-points at baseline, at 2 weeks and at 4 weeks post-treatment. Anaerobic culture methods were used for microbial isolation and identification. The microbiologic examination was carried out blindly. Microbiological data were evaluated by a repeated measures analysis of variance. RESULTS: No statistical difference was found in total colony counts between subgingival sites treated with chlorhexidine chip + Sc/Rp and those treated with Sc/Rp alone. Also, the percentage of major periodontal pathogens (Actinobacillus actinomycetemcomitans, Porphyromonas gingivalis and Bacteroides forsythus) and the percentage of total periodontal pathogens (A. actinomycetemcomitans, P. gingivalis, B. forsythus, Prevotella intermedia-group, Fusobacterium, Eubacterium, Campylobacter rectus, Peptostreptococcus micros, Eikenella corrodens, enteric rods) were not significantly different between the chlorhexidine chip + Sc/Rp group and the Sc/Rp group. At baseline, A. actinomycetemcomitans was recovered from 4 chlorhexidine chip + Sc/Rp sites and 2 Sc/Rp sites, P. gingivalis from 5 chlorhexidine chip + Sc/Rp sites and 4 Sc/Rp sites, and B. forsythus from 9 chlorhexidine chip + Sc/Rp and 7 Sc/Rp sites. At 4 weeks, A. actinomycetemcomitans was detected in 2 chlorhexidine chip + Sc/Rp sites but not in any site receiving Sc/Rp, P. gingivalis in 2 chlorhexidine chip + Sc/Rp sites but not in any Sc/Rp site, and B. forsythus in 1 chlorhexidine chip + Sc/Rp and in 2 Sc/Rp sites. CONCLUSION: The present data obtained from bilateral periodontitis lesions of 13 adults suggest that chlorhexidine chip treatment of adult periodontitis lesions provides little or no additional antimicrobial benefits compared to thorough Sc/Rp alone.

Adult↗

Early detection of spoilage moulds in bread using volatile production patterns and quantitative enzyme assays.

AIMS: Early detection of spoilage fungi (two Eurotium spp., a Penicillium chrysogenum species) in bread analogues over periods of 72 h at 25 degrees C and 0.95 water activity was evaluated using volatile production patterns, hydrolytic enzyme production, and changes in fungal populations. METHODS AND RESULTS: Using an electronic nose system it was possible to differentiate between uninoculated controls and samples contaminated with P. chrysogenum within 28 h. After 40-48 h it was possible to differentiate between the Eurotium spp., P. chrysogenum and the control using Principal Component Analysis (PCA). Cluster analyses could differentiate between the control, P. chrysogenum and the Eurotium spp. after 40 h. Of seven hydrolytic enzymes examined after 48 h, the specific activities of three were significantly different from uninoculated control bread. There were also differences between the mould species in production of three enzymes. Penicillium chrysogenum populations increased fastest, from about 10(3) cfu g(-1) to 10(6)-10(7) cfu g(-1) after 72 h. For the Eurotium spp. this increase was slower. CONCLUSIONS: Overall, this study suggests, for the first time, that an electronic nose system using a surface polymer sensor array is able to detect qualitative changes in volatile production patterns for the early detection of the activity of spoilage moulds in bakery products. SIGNIFICANCE AND IMPACT OF THE STUDY: Potential exists for application of such systems for microbial quality assurance in intermediate moisture food products.

Ascomycota↗

Sonicated vascular catheter tip cultures. Quantitative association with catheter-related sepsis and the non-utility of an adjuvant cytocentrifuge Gram stain.

Microbial quantification of potentially infected intravenous catheters is used to determine whether the removed device may be the source of sepsis. In this study, the authors defined a clinically significant colony count using the sonication technique to implicate the catheter as the source of sepsis. The utility of a cytocentrifuge-prepared Gram's stain of sonication broth as a rapid test for the accurate diagnosis of catheter-related bacteremia and fungemia was also evaluated. Initially, 230 consecutive vascular catheter cultures were performed. In 14 patients, the blood culture was found to grow in the same organism as the catheter culture. The eight blood-culture positive patients with probable catheter-related sepsis had four catheters with 10(4)-10(5), and four with more than 10(5) CFU derived from the catheter sonication broth. Six bacteremias not catheter related showed three catheters with < 10(3), two with 10(3)-10(4), and one with > 10(5) CFU. One catheter-related bacteremia had a negative culture. A subsequent validation study on 175 catheter specimens using 1,000 CFU as a cut-off between a positive and negative test fully separated eight patients with catheter-related bacteremia from six patients with infection at a distant site. Cytospin Gram stain, studied in the initial evaluation, was positive in 17 catheters with < 10(3) CFU, 4 at 10(3)-10(4), 4 at 10(4)-10(5) and 9 with > 10(5) CFU in the sonication broth. No correlation was found between this test and catheter-related sepsis. From these results, the authors concluded that those catheters with counts below 10(3) CFU in the catheter sonication broth do not appear to have an association with catheter-related sepsis, and that a Cytospin Gram stain done with the sonication technique does not correlate with the presence of catheter sepsis.

Bacteremia↗