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L’objectif principal du traitement endodontique est la prévention ou la suppression d’une infection du système canalaire, par l’élimination des micro-organismes le colonisant. Le protocole est parfaitement codifié : préparation mécanique, utilisation de solutions de désinfection et scellement étanche. L’irrigation joue un rôle essentiel dans ce processus de nettoyage puisqu’elle permet le retrait de la boue dentinaire et des bactéries. L’agitation des solutions d’irrigation, aussi appelée activation, permet d ’optimiser leur pouvoir solvant et désinfectant. De nos jours, il existe de nombreuses méthodes d’activation mais aucune ne permet la stérilisation complète du réseau canalaire, notamment au niveau du tiers apical. De plus, les innovations technologiques de mise en forme à l’aide d’instruments rotatifs réduisent le temps de préparation, et donc le temps de contact des solutions d’irrigation avec les parois radiculaires. Le laser, couramment utilisé en endodontie semblerait améliorer la désinfection, en augmentant la pénétration et l’efficacité des solutions d’irrigation. Nous nous proposons ici de présenter son intérêt dans la désinfection canalaire, et de confronter les différentes techniques d’activation. Ainsi, nous tenterons d’évaluer l’apport du laser dans les protocoles actuels et voir comment ils pourraient pallier le faible volume de solution atteignant le tiers apical.

International audience; Legionella pneumophila is frequently detected in hot water distribution systems and thermal control is a common measure implemented by health care facilities. A risk assessment based on water temperature profiling and temperature distribution within the network is proposed, to guide effective monitoring strategies and allow the identification of high risk areas. Temperature and heat loss at control points (water heater, recirculation, representative points-of-use) were monitored in various sections of five health care facilities hot water distribution systems and results used to develop a temperature-based risk assessment tool. Detailed investigations show that defective return valves in faucets can cause widespread temperature losses because of hot and cold water mixing. Systems in which water temperature coming out of the water heaters was kept consistently above 60 °C and maintained above 55 °C across the network were negative for Legionella by culture or qPCR. For systems not meeting these temperature criteria, risk areas for L. pneumophila were identified using temperature profiling and system's characterization; higher risk was confirmed by more frequent microbiological detection by culture and qPCR. Results confirmed that maintaining sufficiently high temperatures within hot water distribution systems suppressed L. pneumophila culturability. However, the risk remains as shown by the persistence of L. pneumophila by qPCR.

International audience; Due to their high surface-volume ratio, their laminar flow and frequent stagnation periods, dental unit waterlines (DUWL) foster the attachment of microorganisms and the development of biofilm, resulting in the continuous contamination of the outlet water from dental units; this contamination may be responsible for a potential risk of infection due to the exposure of patients and medical staff to droplet inhalation or splashed water. In this study, the anti-biofilm activity of three disinfectants recommended by dental unit manufacturers -Calbenium(©), Oxygenal 6(©) and Sterispray(©) - was evaluated. A dynamic model simulating DUWL conditions was developed and polymicrobial biofilms containing bacteria (Pseudomonas aeruginosa), fungi (Candida albicans) and Free Living Amoeba (FLA: Vermamoeba vermiformis) were allowed to form. The ability of disinfectants to reduce biofilm formation or to eradicate an already formed biofilm was evaluated. Results showed the various effects of the tested disinfectants according to their composition, concentration and the targeted species. V. vermiformis was resistant to disinfectants, regardless of the tested concentrations and the concentrations recommended by manufacturers were not the most appropriate. Results also showed that Calbenium(©) was the most effective disinfectant to reduce already formed biofilms; its maximum efficiency was observed from 0.5% on both P. aeruginosa and C. albicans compared to 2 and 3% respectively for Sterispray(©). The maximum efficiency of Oxygenal(©) was observed from 3% on P. aeruginosa but Oxygenal(©) was unable to totally eliminate C. albicans in the tested conditions, contrary to other disinfectants. Calbenium(©) was able to prevent biofilm formation efficiently even if it displayed no prophylactic activity against V. vermiformis. Overall, the FLA survival may contribute to maintaining other species. Finally the tested disinfectants were partially active against sessile microorganisms and more suitable concentrations could be used to increase their efficacy. Their use in a prophylactic rather than curative way should be recommended.

International audience; The HIWATE (Health Impacts of long-term exposure to disinfection byproducts in drinking WATEr) project was a systematic analysis that combined the epidemiology on adverse pregnancy outcomes and other health effects with long-term exposure to low levels of drinking water disinfection byproducts (DBPs) in the European Union. The present study focused on the relationship of the occurrence and concentration of DBPs with in vitro mammalian cell toxicity. Eleven drinking water samples were collected from five European countries. Each sampling location corresponded with an epidemiological study for the HIWATE program. Over 90 DBPs were identified; the range in the number of DBPs and their levels reflected the diverse collection sites, different disinfection processes, and the different characteristics of the source waters. For each sampling site, chronic mammalian cell cytotoxicity correlated highly with the numbers of DBPs identified and the levels of DBP chemical classes. Although there was a clear difference in the genotoxic responses among the drinking waters, these data did not correlate as well with the chemical analyses. Thus, the agents responsible for the genomic DNA damage observed in the HIWATE samples may be due to unresolved associations of combinations of identified DBPs, unknown emerging DBPs that were not identified, or other toxic water contaminants. This study represents the first to integrate quantitative in vitro toxicological data with analytical chemistry and human epidemiologic outcomes for drinking water DBPs.

International audience; Municipal sewage effluents are complex mixtures that are known to compromise the health condition of aquatic organisms. The aim of this study was to evaluate the impacts of various wastewater disinfection processes on the immune system of juvenile rainbow trout (Oncorhynchus mykiss). The trout were exposed to a primary-treated effluent for 28 days before and after one of each of the following treatments: ultraviolet (UV) radiation, ozonation and peracetic acid. Immune function was characterized in leucocytes from the anterior head kidney by the following three parameters: phagocytosis activity, natural cytotoxic cells (NCC) function and lymphocyte (B and T) proliferation assays. The results show that the fish mass to length ratio was significantly decreased for the primary-treated and all three disinfection processes. Exposure to the primary-treated effluent led to a significant increase in macrophage-related phagocytosis; the addition of a disinfection step was effective in removing this effect. Both unstimulated and mitogen-stimulated T lymphocyte proliferation in fish decreased dramatically in fish exposed to the ozonated effluent compared to fish exposed to either the primary-treated effluent or to aquarium water. Stimulation of T lymphocytes proliferation was observed with the peracetic acid treatment group. In conclusion, the disinfection strategy used can modify the immune system in fish at the level of T lymphocyte proliferation but was effective to remove the effects on phagocytosis activity.

International audience; Cleaning products may cause work-related asthma, but information regarding the specific exposures involved is scarce. We aimed to determine the associations between asthma and occupational exposure to cleaning agents in hospital workers. Analyses were conducted in 179 (136 women) hospital workers and a reference population of 545 subjects (18-79 years) from the French case-control and familial Epidemiological study on the Genetics and Environment of Asthma (2003-2007). Exposures to cleaning agents were estimated using three methods: self-report, expert assessment and an asthma-specific job-exposure matrix (JEM). Associations between cleaning products and current asthma were evaluated by logistic regressions, stratified by sex and adjusted for age and smoking status. According to expert assessment, 55% of male and 81% of female hospital workers were exposed to cleaning/disinfecting tasks weekly (p

International audience; Drosophila melanogaster is a robust model to investigate many biological problems. It is however prone to some infections, which may endanger fly stocks if left unchecked for. One such infection is caused by an obligate fungal intracellular parasite, Tubulinosema ratisbonensis, which can be found in laboratory stocks. Here, we identify and briefly characterize a T. ratisbonensis strain that was infesting our Drosophila cultures and that required intensive measures to contain and eradicate the infection. We describe the phenotypes of infested stocks. We also report PCR-based techniques that allow the detection of infested stocks with a high sensitivity. We have developed a high-throughput qPCR assay that allows the efficient parallel screening of a large number of potentially-infested stocks. We also have investigated several prophylactic measures to prevent the further contamination of stocks, namely UV-exposure, ethanol treatment, bleaching, and desiccation. Bleaching was found to kill all spores. Other treatments were less effective but were found to be sufficient to prevent further contamination of noninfested stocks. Two treatments were efficacious in curing infested stocks (1) bleaching of eggs and subsequent raising of the larvae in clean vials; (2) fumagillin treatment. These cures only work on stocks that have not become too weak to withstand the procedures.

Legionella species are frequently detected in hot water systems, attached to the surface as a biofilm. In this work, the dynamics of Legionella spp. and diverse bacteria and eukarya associated together in the biofilm, coming from a pilot scale 1 system simulating a real hot water system, were investigated throughout 6 months after two successive heat shock treatments followed by three successive chemical treatments. Community structure was assessed by a fingerprint technique, single-strand conformation polymorphism (SSCP). In addition, the diversity and dynamics of Legionella and eukarya were investigated by small-subunit (SSU) ribosomal cloning and sequencing. Our results showed that pathogenic Legionella species remained after the heat shock and chemical treatments ( Legionella pneumophila and Legionella anisa , respectively). The biofilm was not removed, and the bacterial community structure was transitorily affected by the treatments. Moreover, several amoebae had been detected in the biofilm before treatments ( Thecamoebae sp., Vannella sp., and Hartmanella vermiformis ) and after the first heat shock treatment, but only H. vermiformis remained. However, another protozoan affiliated with Alveolata, which is known as a host cell for Legionella , dominated the eukaryal species after the second heat shock and chemical treatment tests. Therefore, effective Legionella disinfection may be dependent on the elimination of these important microbial components. We suggest that eradicating Legionella in hot water networks requires better study of bacterial and eukaryal species associated with Legionella in biofilms.

International audience; Water and air quality of fifteen swimming pools using chlorine disinfection was measured during eight sampling campaigns in Rennes, France. Concentrations of trihalomethanes (THMs) and trichloramine (NCl(3)) in air as well as concentrations of parameters, including total organic carbon (TOC), chlorine (free, combined and total), pH, Kjeldhal nitrogen (KN), chloride ions and THMs in water and air temperature, were measured. Water and air samples were collected during 3h in the morning and afternoon, during high and low attendance days, in summer and winter. Data analysis was perfomed by multivariate ordinary least square (OLS) and quantile (QR) regressions, from both data quality and other parameters such as the ratio between the number of visitors and the volume of water available V/W, the proportion of air re-circulated and the number of swimmers during sampling. The geometric means (GM±95% CIs) of the sum of THMs (TTHM) and NCl(3) in air were 74.9±6.7 μg/m(3) and 190±10 μg/m(3), respectively. As expected, concentrations of TTHM in air were correlated to concentrations of TTHM in water. Concentrations of TTHM and NCl(3) in air measured in winter were significantly higher than in summer, depicting that low air exchange rate in winter lead to accumulation of pollutants. NCl(3) (especially at higher quantiles) and TTHM concentrations in air were associated to the number of swimmers. NCl(3) was also linked to air temperature and pH. Moreover TTHM and NCl(3) in air seemed to be affected by additional determinants including the volume of hall and the ratio V/W. In a process of reducing exposure, the OLS regression provides useful information on key parameters influencing concentrations of THMs and NCl(3) in indoor swimming pools. In addition, QR allows identifying factors associated to elevated levels of contamination.

International audience; Several epidemiological studies suggested an association between the risk of bladder cancer and the exposure to trihalomethanes (THMs), the main disinfection by-products (DBPs) of chlorinated water. A previous pooled analysis of case-control studies from North America and Europe estimated a summarized dose-response relation. For policy guidance of drinking water disinfection in Europe and because major differences exist in water disinfection practices and DBPs occurrence between both continents, specific risk estimates for bladder cancer in relation to DBPs exposure for European populations were needed. We conducted a pooled and a two-stage random-effect meta-analyses of three European case-control studies from France, Finland, and Spain (5467 individuals: 2381 cases and 3086 controls). Individual exposure to THMs was calculated combining information on residential history, estimates of the average total THMs (TTHM) level in tap water at the successive residences and personal water consumption. A significant odds-ratio was observed for men exposed to an average residential TTHM level > 50 μg/l (OR = 1.47 (1.05; 2.05)) when compared to men exposed to levels ≤ 5 μg/l. The linear trend of the exposure-risk association was significant (p = 0.01). Risks increased significantly for exposure levels above 25 μg/l and with more than 30 years of exposure to chlorinated water, but were mainly driven by the level rather than the duration of exposure. No significant association was found among women or with cumulative exposure through ingestion. There was no evidence of a differential exposure-response relation for TTHM and bladder cancer in Europe and North America. Consequently, a global exposure-risk relation based on 4351 cases and 7055 controls is now available.

International audience; Biofilm formation is a key step during marine biofouling, the natural colonization of immersed substrata, leading to major economic and ecological consequences. Consequently, bacteria have been used for the screening of new non-toxic antifoulants: the adhesion of five strains isolated on three French locations was monitored using a fluorescence-based assay and toxicity was also evaluated. Nine biocides including commercial, natural and natural-derived products were tested. The commercial antifoulants, TBTO and Sea Nine showed low EC(50) but high toxicity. The non-commercial products TFA-Z showed significant anti-adhesion activities and appeared to be non-toxic, suggesting a specific anti-adhesion mechanism. In addition, the strains could be classified depending on their sensitivity to the molecules used even if strain sensitivity also depended on the molecules tested. In conclusion, TFA-Z would be a promising candidate as non-toxic antifoulant and our results strengthen the need to perform antifouling bioassays with a panel of strains showing different response profiles.

International audience; Silica and paramagnetic silica microparticles are surface-modified by an antibacterial macromolecular coating. For this, a hydrophilic copolymer brush based on oligo(ethylene glycol) methacrylates is grown on the particle surface by surface-initiated ATRP. Then, Magainin-I, a natural antimicrobial peptide, is grafted onto the hydroxyl groups of the brush through a heterolinker. The grafting of the peptide is evidenced by fluorescence microscopy and X-ray photoelectron spectroscopy. Moreover, culturability and viability assays performed in the presence of the magainin-grafted particles prove their bactericidal properties. The rapid recovery of the bactericidal particles based on paramagnetic silica and suspended in solution is shown under magnetization. Such particles offer the advantage to treat efficiently various sensitive aqueous solutions while avoiding any dissemination of bactericidal substances in the environment. As a consequence, they are of a great interest for various applications in medical, cosmetic, or biomedical fields.

International audience; OBJECTIVES: The aim of this study is to evaluate the quality of disinfection of endoscopes at Brest hospital over a period from 2007 to 2009. PATIENTS AND METHODS: Retrospective study of microbiological investigations of endoscopes done at Brest hospital from 2007 to 2009. The interpretation of the microbiological investigations is based on the recommendations of the Comité technique national des infections nosocomiales et infections liées aux soins (CTINILS) of 2007. RESULTS: Most of the controls realized over the period deal with gastroenterological endoscopes (63.4 %) and bronchial endoscopes (21.8 %). Most of the controls (66.8 %) are conformed to the target level. Only 26.7 % of the controls get the level of action. Globally, the rate of level of action significantly increases (p=0.004) from 2007 (21.2 %) to 2009 (35.6 %). This increase is relatively important in gastroenterology endoscopy (46.8 % in 2009 versus 24.1 % in 2007) whereas the rate decreases in bronchial endoscopy (14.8 % in 2009 versus 25.9 % in 2007). In gastroenterological endoscopy, rates vary with the type of endoscopes and the context of controls, but there is no significant difference between manual disinfection and automated disinfection. The most frequent germ found in gastroenterological and bronchial endoscopies is Pseudomonas aeruginosa. CONCLUSION: Our results show that it is very difficult to insure a perfect disinfection of endoscopes. Difficulties met are certainly related with the complexity of the endoscopes and of the techniques of disinfection. Infections of patients are very infrequent in endoscopy, which takes the question of the pertinence of the threshold used for microbiological investigations.

A novel thermophilic, anaerobic and organotrophic bacterium, designated strain MC3T, was isolated from a coastal thermal spring on Île Saint-Paul in the Southern Indian Ocean. Cells of strain MC3T were motile rods, 0.8–1.0 μm wide and 1.0–2.4 μm long during exponential phase and up to 7.0 μm long during stationary phase. Strain MC3T was an anaerobic organotroph able to use diverse organic compounds. It was also able to reduce sulfur to sulfide. Growth was observed at temperatures ranging from 45 to 70 °C (optimum at 60 °C), between pH 5.5 and 7.5 (optimum at pH 6) and from 8 to 46 g NaCl l−1 (optimum at 26 g l−1). The total G+C content of the genomic DNA was 26.2 mol%. Phylogenetic analysis based on 16S rRNA gene sequence comparisons indicated that strain MC3T was affiliated with the genus Marinitoga within the order Thermotogales. It shared 94.4–95.7 % 16S rRNA gene sequence similarity with strains of other Marinitoga species; Marinitoga hydrogenitolerans was found to be the most closely related organism. Based on the data from the phylogenetic analysis and the physiological properties of the novel isolate, strain MC3T should be classified as a representative of a novel species, for which the name Marinitoga litoralis sp. nov. is proposed; the type strain is MC3T (=DSM 21709T =JCM 15581T).

International audience; We examined the variations of bacterial populations in treated drinking water prior to and after the final chlorine disinfection step at two different surface water treatment plants. For this purpose, the bacterial communities present in treated water were sampled after granular activated carbon (GAC) filtration and chlorine disinfection from two drinking water treatment plants supplying the city of Paris (France). Samples were analyzed after genomic DNA extraction, polymerase chain reaction (PCR) amplification, cloning, and sequencing of a number of 16S ribosomal RNA (rRNA) genes. The 16S rDNA sequences were clustered into operational taxonomic units (OTUs) and the OTU abundance patterns were obtained for each sample. The observed differences suggest that the chlorine disinfection step markedly affects the bacterial community structure and composition present in GAC water. Members of the Alphaproteobacteria and Betaproteobacteria were found to be predominant in the GAC water samples after phylogenetic analyses of the OTUs. Following the chlorine disinfection step, numerous changes were observed, including decreased representation of Proteobacteria phylotypes. Our results indicate that the use of molecular methods to investigate changes in the abundance of certain bacterial groups following chlorine-based disinfection will aid in further understanding the bacterial ecology of drinking water treatment plants (DWTPs), particularly the disinfection step, as it constitutes the final barrier before drinking water distribution to the consumer's tap.

International audience; In the framework of a cohort study of pregnant women conducted in Brittany (France), we assessed the exposure to trihalomethanes (THM) during pregnancy in a subset by evaluating (1) potential sources of variability in household THM levels; (2) the between- and within-subject variability in THM levels; (3) THM levels in swimming pools; and (4) the role of water-related habits on total THM uptake. We visited 109 women from the ongoing cohort study at home for an interview and collection of tap water from October to December 2004. Forty-three of them were re-contacted to obtain a second tap water sample in April-May 2005. We designed a questionnaire to collect individual information on source and amount of drinking water, frequency of showering, bathing, and swimming pool attendance, and household characteristics. We obtained 282 THM measurements, 152 specifically for the study and 130 from a regulatory agency. Personal information and environmental data were combined using two methodologies (method 1 using regulatory data and method 2 using our THM measurements) with a different set of assumptions. We calculated ingestion, showering, bathing, and swimming pool THM uptakes and added up those uptakes to calculate total THM uptake. Average THM levels from our measurements in October, November-December, and April-May were 61.3, 45.1, and 54.5 microg/l, respectively. Geographical variability was low and characteristics of the household did not influence THM levels. Within-subject variability in THM levels was three times higher than between-subject variability. Average THM level in swimming pools was 80.4 microg/l. Average water consumption during pregnancy was 1.9l/day. The source of the household drinking water was 90% bottled, 8% municipal, and 2% from other sources. Forty-seven per cent attended swimming pools during pregnancy. Using method 1, the geometric mean of total THM uptake was 0.93 microg/day. Showering contributed 64%, swimming in pools 23%, bathing 12%, and drinking water 1% to the total THM uptake. In a setting with low geographical variability and limited environmental measurements, individual data is highly relevant to determine personal THM exposure and uptake. In a population that mainly drinks bottled water (e.g., pregnant women), individual THM uptakes are dominated by inhalation and dermal absorption during, showering, swimming in pools, and bathing.

BACKGROUND: Most studies of corneal donor contamination concentrate on postenucleation contamination of the eyeball. The purpose of the present study was to evaluate the relative contamination of in situ excised corneal tissue and relevance to final success or rejection by recipients of the corneal grafts. METHODS: Ninety-three donors underwent anterior chamber puncture (ACP) and corneal epithelium scarification (CS) before and after disinfection with 5% povidone-iodine. Following in situ excision, corneas were preserved in culture medium at +35 degrees C. Morphological and bacteriological assessment was carried out after culture, and recipients were followed up over a 2-year period. RESULTS: Samples taken by ACP, CS before disinfection, CS after disinfection and a culture medium sample were contaminated by bacteria in, respectively, 8 (8.6%), 23 (24.7%) 4 (4.3%) and 5 (5.4%) donors. Contamination of aqueous humor was significantly associated with age, death-to-sample time and premortem systemic infection. Contamination of epithelium significantly increased culture medium contamination. CONCLUSION: External bacteria on donor cornea are mainly skin bacteria (especially Staphylococcus) and can be partially eliminated by a povidone-iodine wash. Internal bacteria are mainly gut bacteria and may be due to perimortem bacteriemia. However, bacterial infection at the time of death appears to have no effect on the incidence of endophthalmitis in recipients and should no longer prevent use of such corneal tissue in grafts.