Mycobacterium lutetiense sp. nov., Mycobacterium montmartrense sp. nov. and Mycobacterium arcueilense sp. nov., members of a novel group of non-pigmented rapidly growing mycobacteria recovered from a water distribution system.

From our recent survey of non-pigmented rapidly growing mycobacteria in the Parisian water system, three groups of isolates (taxons 1-3) corresponding to possible novel species were selected for taxonomic study. The three taxa each formed creamy white, rough colonies, had an optimal growth temperature of 30 °C, hydrolyzed Tween 80, were catalase-positive at 22 °C and expressed arylsulfatase activity. All three were susceptible to amikacin, ciprofloxacin and tigecycline. The three taxa produced specific sets of mycolic acids, including one family that has never previously been described, as determined by thin layer chromatography and nuclear magnetic resonance. The partial rpoB sequences (723 bp) showed 4-6 % divergence from each other and more than 5 % differences from the most similar species. Partial 16S rRNA gene sequences showed 99 % identity within each species. The most similar sequences for 16S rRNA genes (98-99 % identity over 1444-1461 bp) were found in the Mycobacterium fortuitum group, Mycobacterium septicum and Mycobacterium farcinogenes. The three taxa formed a new clade (bootstrap value, 99 %) on trees reconstructed from concatenated partial 16S rRNA, hsp65 and rpoB sequences. The above results led us to propose three novel species for the three groups of isolates, namely Mycobacterium lutetiense sp. nov. [type strain 071T=ParisRGMnew_1T (CIP 110656T=DSM 46713T)], Mycobacterium montmartrense sp. nov. [type strain 196T=ParisRGMnew_2T (CIP 110655T=DSM 46714T)] and Mycobacteriu marcueilense sp. nov. [type strain of 269T=ParisRGMnew_3T (CIP 110654T=DSM 46715T)].

Perchlorate: water and infant formulae contamination in France and risk assessment in infants.

Perchlorate ions ClO4(-), known to inhibit competitively the uptake of iodine by the thyroid, have been detected in drinking water in France as well as in infant formulae. A tolerable daily intake (TDI) has been established at 0.7 µg kg(-1) bw day(-1) based on the inhibition of iodine uptake. Due to this mechanism of action, the iodine status could strongly influence the biological effect of perchlorate. Perchlorate concentrations in water and infant formulae were measured and the exposure of children under 6 months of age calculated. It appeared that the TDI could be exceeded in some children. As the iodine status is not optimal within the entire French population, there appears to be a need to clarify the sources of perchlorate ultimately to decrease exposure.

First evidence of amoebae-mycobacteria association in drinking water network.

Free-living amoebae are protozoa ubiquitously found in water systems. They mainly feed on bacteria by phagocytosis, but some bacterial species are able to resist or even escape this lethal process. Among these amoeba resistant bacteria are numerous members of the genus Mycobacterium. Nontuberculous Mycobacteria (NTM) are opportunistic pathogens that share the same ecological niches as amoebae. While several studies have demonstrated the ability of these bacteria to colonise and persist within drinking water networks, there is also strong suspicion that mycobacteria could use amoebae as a vehicle for protection and even replication. We investigated here the presence of NTM and FLA on a drinking water network during an all year round sampling campaign. We observed that 87.6% of recovered amoebal cultures carried high numbers of NTM. Identification of these amoeba and mycobacteria strains indicated that the main genera found in drinking water networks, that is, Acanthamoeba, Vermamoeba, Echinamoeba, and Protacanthamoeba are able to carry and likely to allow replication of several environmental and potentially pathogenic mycobacteria including M. llatzerense and M. chelonae. Direct Sanger sequencing as well as pyrosequencing of environmental isolates demonstrated the frequent association of mycobacteria and FLA, as they are part of the most represented genera composing amoebae's microbiome. This is the first time that an association between FLA and NTM is observed in water networks, highlighting the importance of FLA in the ecology of NTM.

Variations in trihalomethane levels in three French water distribution systems and the development of a predictive model.

Epidemiological studies have demonstrated that chlorination by-products in drinking water may cause some types of cancer in humans. However, due to differences in methodology between the various studies, it is not possible to establish a dose-response relationship. This shortcoming is due primarily to uncertainties about how exposure is measured-made difficult by the great number of compounds present-the exposure routes involved and the variation in concentrations in water distribution systems. This is especially true for trihalomethanes for which concentrations can double between the water treatment plant and the consumer tap. The aim of this study is to describe the behaviour of trihalomethanes in three French water distribution systems and develop a mathematical model to predict concentrations in the water distribution system using data collected from treated water at the plant (i.e. the entrance of the distribution system). In 2006 and 2007, samples were taken successively from treated water at the plant and at several points in the water distribution system in three French cities. In addition to the concentrations of the four trihalomethanes (chloroform, dichlorobromomethane, chlorodibromomethane, bromoform), many other parameters involved in their formation that affect their concentration were also measured. The average trihalomethane concentration in the three water distribution systems ranged from 21.6 µg/L to 59.9 µg/L. The increase in trihalomethanes between the treated water at the plant and a given point in the water distribution system varied by a factor of 1.1-5.7 over all of the samples. A log-log linear regression model was constructed to predict THM concentrations in the water distribution system. The five variables used were trihalomethane concentration and free residual chlorine for treated water at the plant, two variables that characterize the reactivity of organic matter (specific UV absorbance (SUVA), an indicator developed for the free chlorine consumption in the treatment plant before distribution δ) and water residence time in the distribution system. French regulations impose a minimum trihalomethane level for drinking water and most tests are performed on treated water at the plant. Applied in this context, the model developed here helps better to understand trihalomethane exposure in the French population, particularly useful for epidemiological studies.

Variations of bacterial 16S rDNA phylotypes prior to and after chlorination for drinking water production from two surface water treatment plants.

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.

Assessment of phylogenetic diversity of bacterial microflora in drinking water using serial analysis of ribosomal sequence tags.

We examined chlorinated drinking water samples from three different surface water treatment plants for bacterial 16S rDNA diversity using the serial analysis of V6 ribosomal sequence tag (SARST-V6) method. A considerable degree of diversity was observed in each sample, with an estimated richness ranging from 173 to 333 phylotypes. The community structure shows that there are differences in bacterial evenness between sampled sites. The taxonomic composition of the microbial communities was found to be dominated by members of the Proteobacteria (57.2-77.4%), broadly distributed among the classes Alphaproteobacteria, Betaproteobacteria, Gammaproteobacteria, and Deltaproteobacteria. Additionally, a large proportion of sequences (6.3-36.5%) were found to be distantly related to database sequences of unknown phylogenetic affiliation. Given the apparent significance of this bacterial group in drinking water, a 16S rDNA analysis was performed and confirmed their presence and phylogeny. Notwithstanding the potential under-representation of certain bacterial phyla using the SARST-V6 primer pairs, as revealed by a refined computer algorithm, our results suggest that 16S rDNA corresponding to a variety of eubacterial groups can be detected in finished drinking water, suggesting that this water may contain a higher level of bacterial diversity than previously observed.

Effects of ozone and ultraviolet radiation treatments on the infectivity of Toxoplasma gondii oocysts.

Clinical toxoplasmosis in humans has been epidemiologically linked to the consumption of drinking water contaminated by Toxoplasma gondii oocysts. We evaluated killing of T. gondii oocysts after ultraviolet (UV) or ozone treatments by bioassay in mice and/or cell culture. A 4-log inactivation of the oocyst/sporozoite infectivity was obtained for UV fluences >20 mJ cm(-2). In contrast, oocysts were not inactivated by ozone with an exposure (Ct) up to 9.4 mg min l (-1) in water at 20 degrees C. In conclusion, UV treatment can be an effective disinfection method to inactivate T. gondii oocysts in drinking water, but ozone did not show promise in this research.

Genotoxicity of organic extracts of urban airborne particulate matter: an assessment within a personal exposure study.

Airborne particulate matter, PM(10) and PM(2.5), are associated with a range of health effects including lung cancer. Their complex organic fraction contains genotoxic and carcinogenic compounds such as polycyclic aromatic hydrocarbons (PAHs) and their derivatives. This study evaluates the genotoxicity of the PM(10) and PM(2.5) organic extracts that were sampled in the framework of a personal exposure study in three French metropolitan areas (Paris, Rouen and Strasbourg), using the comet assay, performed on HeLa S3 cells. In each city, 60-90 non-smoking volunteers composed of two groups of equal size (adults and children) carried the personal Harvard Chempass multi-pollutant sampler during 48h along two different seasons ('hot' and 'cold'). Volunteers were selected so as to live (home and work/school) in 3 different urban sectors contrasted in terms of air pollution within each city (one highly exposed to traffic emissions, one influenced by local industrial sources, and a background urban environment). Genotoxic effects are stronger for PM(2.5) extracts than for PM(10), and greater in winter than in summer. Fine particles collected by subjects living within the traffic proximity sector present the strongest genotoxic responses, especially in the Paris metropolitan area. This work confirms the genotoxic potency of particulate matter (PM(10) and PM(2.5)) organic extracts to which urban populations are exposed.

Carbon disulfide cytotoxicity on cultured cardiac myocyte cell of rats.

Although mostly epidemiological studies suggested that carbon disulfide produces cardiovascular effects in occupationally exposed workers, little is known about its cellular mechanism. The purpose of the present study was to evaluate the functional and histological effects on cardiac myocytes cultured under the condition of carbon disulfide exposure. Cardiac myocytes were isolated from neonatal rat ventricles by trypsin dispersion and cultured for 3 days in a full (Dulbecco's modified eagle medium) medium containing 2% calf serum. Thereafter the myocytes (10(6) myocytes/culture flask) were incubated with carbon disulfide at (CS(2)) the concentrations of 0, 20, 40, and 80 micromol/mL) for 24h. The beating arrest rate of myocytes for each group was examined and succinodehydrogenase (SDH) activity in the myocardial cells was also assessed by cytochemical method, and morphological examination was also performed. We found that the beating arrest rate of cardiac myocytes increased with increasing exposure levels. Vacuolization and pseudopodia may be seen in the cytoplasm of exposure group. SDH activity decreased with increasing exposure levels. The results suggested that CS(2) has a direct cytotoxic effect which is dose dependent. The biochemical mechanism may be a reduction of the availability of energy of the cardiac cytocyte in the form of ATP, resulting in a decrease of contractility by lacking of energy.

Cardiovascular effects of carbon disulfide: meta-analysis of cohort studies.

Objectives were to review the evidence of cardiovascular effects of carbon disulfide (CS2) on exposed workers from different cohort studies. Databases searched included MEDLINE, TOXLINE, and OSHtM Database (CD-ROM). Reference lists of reports and reviews were also searched. To be included in the analysis, a study had to have been a cohort study on cardiovascular effects of carbon disulfide with at least one outcome of interest. Every report was independently read using the Lichtenstein scale, General information of each study and relative risk estimates were calculated with 95% confidence intervals using the fixed-effects model. Pooled relative risks were calculated by the Dersimonian-Laird method basing on the random-effects model. Eleven cohort studies were identified in an extensive literature review. Pooled RR value was 1.56 and its 95% confidence interval was from 1.12 to 2.1, which showed CS2 exposure has feeble correlation with the prevalence of cardiovascular diseases. It is proved that meta-analysis is a sensitive method for combining different results in occupational epidemiology.

Carbon disulfide cytotoxicity on cultured cardiac myocytes of rats.

Although epidemiological studies have suggested that carbon disulfide produces cardiovascular effects in occupationally exposed workers, little is known about its cellular mechanism. The purpose of the present study was to evaluate the functional and histological effects on cardiac myocytes cultured under a condition of carbon disulfide exposure. Cardiac myocytes were isolated from neonatal rat ventricles by trypsin, dispersed and cultured for 3 days in a full Dulbecco's Modified Eagle Medium containing 2% calf serum. Then the myocytes (10(6) myocytes/mL) were incubated with carbon disulfide at concentrations of 0, 20, 40, and 80 micromol/mL for 24 h. The beating arrest rate of myocytes for each group was examined, succinodehydrogenase (SDH) activity in the myocardial cells was assessed using a cytochemical method, and a morphological examination was performed. We found that the beating arrest rate of cardiac myocytes increased with increasing exposure levels. Vacuolization and pseudopodia could be seen in the cytoplasm of the exposed group. SDH activity decreased with increasing exposure levels. The results suggest that CS2 has a direct and dose-dependent cytotoxic effect. The biochemical mechanism may be a reduction of the availability of energy (adenosine triphosphate) to cardiac myocytes, resulting in a decrease of contractility by lack of energy.

Recherche des principes actifs de Rosmarinus officinalis / Research of the active principles of Rosmarinus officinalis

Le romarin, Rosmarinus officinalis, est traditionnellement utilise dans les affections de la sphere hepatobiliaire sous forme d'infusion aqueuse ou de solutions hydro-alcooliques de degres divers. Il fait partie de la famille des Lamiacees dont plusieurs representants semblent presenter des proprietes antioxydantes particulierement interessantes. Des travaux recents ont montre, sur des modeles in vitro, que le romarin s'opposait aux consequences toxiques de la production de radicaux libres et notamment aux desordres membranaires provoques par la peroxydation lipidique