Search

Les performances atteintes par les deux systemes biologiques etudies – boues activees conventionnelles (BA) et reacteur biologique a biomasse fluidisee (MBBR) – dans le traitement des dix micro— polluants d’origine pharmaceutique quantifies sont similaires et cela grâce au niveau d’epuration identique en nitrification totale.Les abattements quantifies sont variables et atteignent en moyenne pour les sept composes refractaires seulement 24% (carbamazepine, sulfamethoxazole, propranolol, atenolol, diclofenac, econazole, acide salicylique). Les trois autres composes, bien elimines en traitement biologique, atteignent en moyenne 96% (paracetamol, ketoprofene et ibuprofene).Le traitement biologique combine a l’ozonation tertiaire elimine efficacement les composes pharmaceutiques de l’eau des l’utilisation des doses faibles d’ozone (0,45 gO3/g de carbone organique dissous COD). Un abattement moyen de 92% d’efficacite est quantifie sur la moyenne de composes pharmaceutiques dissous, a comparer aux 24% a 45% d’abattement obtenu en traitement bio logique seul. Le passage a une dose d’ozone de 1,30 gO3/gCOD permet d’augmenter de 6% l’abattement moyen des micropolluants. Cette performance n’est pas amelioree par l’utilisation des doses d’ozone plus importantes.Contrairement aux tests d’ecotoxicite normalises (peu sensibles pour discriminer des effets toxiques dans les eaux traitees biologiquement ou chimiquement), les effets mesures sur les embryons du poisson Danio rerio comme la mortalite, les anomalies sur le developpement et la genotoxicite ont montre un effet toxique (nul a faible) pour les effluents biotraites ou ozones. Par rapport au traitement biologique, l’ozonation tertiaire ameliore significativement le potentiel de perturbation endocrinienne des effluents par la reduction complete de l’activite oestrogenique et glucocorticoide des effluents.L’ozonation n’augmente ni ne diminue le niveau de toxicite des effluents deja atteint avec le traitement biologique seul. Il existe un bruit persistant probablement lie a la presence d’autres micro - polluants remanents dans l’eau et non quantifies par la methode d’analyse ciblee. Leur activite peut etre a l’origine du niveau d’effet toxique faible obtenu pour certains biomarqueurs. Ces resultats montrent la necessite d’ameliorer la sensibilite des bioessais dans le but d’optimiser l’evaluation des effets ecotoxicologiques residuels dans les differents types d’effluents.L’obtention des signatures chimiques, au travers d’une approche non ciblee, s’avere un outil puissant et complementaire. Elles revelent la complexite des effluents etudies par le nombre important de signaux obtenus. Les cartographies realisees, par l’analyse en composantes principales, permettent d’avoir une vision globale des effluents et de leur mise en relation respective. Cette approche globale complete efficacement les resultats obtenus par le dosage cible des micropolluants.

International audience; Toxicity of wastewater treatment plant (WWTP) effluents is usually assessed with standardized bioassays, as e.g. the Fish Embryo Toxicity test (FET), but assessment of sub-lethal toxic effects requires to develop more adapted tests. The present work aimed to add the recording of several sub-lethal endpoints in exposed zebrafish embryo-larval stages in order to increase the sensitivity of residual toxicity evaluation of WWTP effluents using a semi-quantitative method (iFET score). This approach was complemented with the genotoxicity assessment on the exposed embryos, a sub-acute hazard particularly relevant to evaluate a potential chronic toxicity risk of low and multi-contaminated environmental matrices. Additionally, endocrine activities of effluents were quantified using human reporter cell lines. This test battery was applied to the assessment of the residual toxicity of five biological treatment effluents, further treated or not using various ozonation treatments intended to improve pharmaceutical compounds removal. Acute toxicity towards zebrafish embryos was very low. However, iFET score approach proposed was able to reveal residual toxicity through the presence of developmental abnormalities in all samples tested. Additionally, a low residual genotoxicity was measured in embryos exposed to two of the WWTP effluents when all excepted one exhibited a residual endocrine activity potential in the ng/L range providing complementary information on the occurrence of endocrine active chemicals and their reduction by different processes. To sum up, such a simplified and ecologically relevant test battery was found sensitive enough to characterize and differentiate various residual effluent’s ecotoxicity at contaminant levels of environmental concern.

International audience; Standardized ecotoxicity bioassays show some limits to assess properly long-term residual toxicity of complex mixture of pollutants often present at low concentration, such as stormwaters. Among invertebrate organisms used for ecotoxicity testing, the microcrustacean Ceriodaphnia dubia (C. dubia) is considered as one of the most sensitive, especially regarding reproduction impairment as a toxicity endpoint. Consequently, the present work explores the interest to perform a multi-generational assay based on the study of the reproduction of C. dubia to assess long-term ecotoxicity of complex mixture, using stormwater samples. With this in mind, a battery of standardized bioassays (Daphnia magna mobility, Pseudokirchneriella subcapitata population growth, Heterocypris incongruens growth and one generation C. dubia reproduction inhibition assays) was performed in parallel to a three generation C. dubia reproduction inhibition assay on 2 stormwater samples. Results highlighted that while all standardized bioassays failed to reveal residual toxicity in the stormwater samples, the C. dubia multi-generational assay exhibited an higher sensitivity than the previous ones. No adverse effect was observed for the first exposed generation, but an increase in mortality and a reproduction disturbance was obtained in the second and third exposed generation depending of the sample. Further experiments are now needed to optimize the exposure protocol of this multi-generational assay.

The Cueva del Azufre in Tabasco, Mexico, is a nutrient-rich cave and its inhabitants need to cope with high levels of dissolved hydrogen sulfide and extreme hypoxia. One of the successful colonizers of this cave is the poeciliid fish Poecilia mexicana, which has received considerable attention as a model organism to examine evolutionary adaptations to extreme environmental conditions. Nonetheless, basic ecological data on the endemic cave molly population are still missing; here we aim to provide data on population densities, size class compositions and use of different microhabitats. We found high overall densities in the cave and highest densities at the middle part of the cave with more than 200 individuals per square meter. These sites have lower H2S concentrations compared to the inner parts where most large sulfide sources are located, but they are annually exposed to a religious harvesting ceremony of local Zoque people called La Pesca. We found a marked shift in size/age compositions towards an overabundance of smaller, juvenile fish at those sites. We discuss these findings in relation to several environmental gradients within the cave (i.e., differences in toxicity and lighting conditions), but we also tentatively argue that the annual fish harvest during a religious ceremony (La Pesca) locally diminishes competition (and possibly, cannibalism by large adults), which is followed by a phase of overcompensation of fish densities.

Molluscan species can be affected by various anthropogenic substances. Yet, these effects are disregarded in chemical risk assessment as molluscs are unrepresented in standard OECD guidelines. The project "validation of a mollusc reproduction test" (Federal Environment Agency, code 371165417) deals with the development of a test method with the mudsnail Potamopyrgus antipodarum for OECD purposes. In this context, the influence on reproduction of both, different media and varying snail density, has been observed in independent experiments. Further, the impact of density on the outcome of subsequent cadmium (Cd) toxicity in a test has been investigated to refine the existing methodology. First, adult snails were kept in different test media for 12 weeks. Second, snail density was increased for 4 weeks to induce stress. Snails from each density scenario were used for another 4 weeks in a reproduction test at an equal density with 12 μg Cd/L, respectively. Significant differences in reproduction between medium groups were noted after 4 and 8, but not 12, weeks. Further, reproduction was significantly altered by snail density in the beakers but after subsequent 4 weeks at a constant density, no differences were observed between control groups. Cd reduced reproduction and this effect increased with snail density in the pre-exposure period, demonstrating that a previous stress factor may result in increased sensitivity to chemicals and underlines the need for more standardized breeding conditions to minimize effect variations. Based on the outcome of this study, an acclimatization period of 12 weeks must be guaranteed for specimens transferred to another medium. Further, 4 weeks of acclimatization are necessary after density stress. An additional 12 weeks density experiment showed that medium volume in each replicate can be decreased by half to save on chemicals, water and space during tests.

International audience; Wastewater treatment plants (WWTPs) are one of the main sources of freshwater pollution eventually resulting in adverse effects in aquatic organisms. Treated effluents can contain many micropollutants at concentrations often below the limit of chemical quantification. On a regulatory basis, WWTP effluents have to be non-toxic to the aquatic environment, wherefore not only chemical abatement but also ecotoxicological evaluation through relevant bioassays is required. Standardized bioassays currently used are often not sensitive enough to reveal a residual toxicity in treated effluents. Therefore, attention must be paid to the development of better-adapted approaches implementing more sensitive organisms and relevant endpoints. In this study, the toxicity of two differently treated effluents (activated sludge treated effluents with and without ozonation) towards the ecologically relevant species Gammarus fossarum was evaluated. Organism fitness traits such as reproduction and sperm DNA integrity were followed in exposed organisms. In complement, enzymatic biomarkers were measured indicating the presence of neurotoxic compounds (acetylcholinesterase activity), the presence of pathogens likely to increase the toxic effects of chemical compounds (phenol-oxidase activity), and the presence of toxic compounds inducing detoxification mechanisms (glutathione-S-transferase activity). Enzymatic activities were not modified, but significant sub-lethal effects were observed in exposed organisms. In both effluents, females showed a retarded molt cycle, a reduced fecundity and fertility, and > 90% of developed embryos exhibited developmental malformations. In addition, a slight but significant genotoxic effect was measured in gammarid sperm. In a whole, no difference in toxicity was found between both effluents. Coupling reproduction impairment and genotoxicity assessment in Gammarus fossarum seems to be a valuable and sensitive tool to reveal residual toxicity in effluents containing a mixture of micropollutants at very low concentrations. Finally, a direct relationship between the observed toxic responses and the quantified micropollutant concentrations could not be evidenced.

Extreme environments are often considered a predation refuge for organisms living in them. In southern Mexico several species of poeciliid fishes are undergoing incipient speciation in a variety of extreme (i.e. permanently dark and/or sulphidic) freshwater systems, and previous research has demonstrated reproductive isolation between populations from sulphidic and adjacent benign habitats. In the present study, we investigated bird predation rates (measured as successful captures per minute) in two sulphidic surface and several benign surface habitats, to test the hypothesis that extreme habitats are predation refuges. We found capture rates to be approximately 20 times higher in sulphidic environments: probably facilitated by extremophile poeciliids spending most of their time at the water surface, where they engage in aquatic surface respiration as a direct response to hypoxia. Even birds that are usually not considered major fish predators regularly engage in fish predation in the toxic habitats of southern Mexico. Our results demonstrate that extreme environments do not necessarily represent a refuge from predation, and we discuss the general importance of predation in driving incipient speciation in these systems. Finally, we hypothesize that natural selection via avian predation may play an important role in maintaining reproductive isolation between divergent poeciliid populations. © 2010 The Linnean Society of London, Biological Journal of the Linnean Society, 2010, 101, 417–426.

International audience; A mixture of urban and hospital effluents (50 % v/v) was evaluated for ecotoxicity with an advanced bioassay battery. Mixed effluents were tested before any treatment, af- ter biological treatment alone, and after biological treatment followed by a tertiary ozonation (15 mg O3/L). Laying a high value on the continuance of organisms’ fitness, essential to preserve a healthy receiving ecosystem, the main objective of this study was to combine normalized bioassays with newly developed in vivo and in vitro tests in order to assess alteration of embryo development, growth and reproduction, as well as genotoxic effects in aquatic organisms exposed to complex wastewater effluents. Comparison of the bioassays sensitivity was considered. Contrary to the lack of toxicity observed with normalized ecotoxicity tests, endpoints measured on zebrafish embryos such as developmental abnormalities and genotoxicity demonstrated a residual toxicity in wastewater both after a biological treatment followed or not by a tertiary O3 treatment. However, the ozonation step allowed to alleviate the residual endocrine disrupting potential measure in the bi- ologically treated effluent. This study shows that normalized bioassays are not sensitive enough for the ecotoxicological evaluation of wastewaters and that there is a great need for the development of suitable sensitive bioassays in order to characterize properly the possible residual toxicity of treated effluents.

International audience; The fish embryo toxicity (FET) biotest has gained popularity as one of the alternative approaches to acute fish toxicity tests in chemical hazard and risk assessment. Despite the importance and common acceptance of FET, it is still performed in multiwell plates and requires laborious and time-consuming manual manipulation of specimens and solutions. This work describes the design and validation of a microfluidic Lab-on-a-Chip technology for automation of the zebrafish embryo toxicity test common in aquatic ecotoxicology. The innovative device supports rapid loading and immobilization of large numbers of zebrafish embryos suspended in a continuous microfluidic perfusion as a means of toxicant delivery. Furthermore, we also present development of a customized mechatronic automation interface that includes a high-resolution USB microscope, LED cold light illumination, and miniaturized 3D printed pumping manifolds that were integrated to enable time-resolved in situ analysis of developing fish embryos. To investigate the applicability of the microfluidic FET (μFET) in toxicity testing, copper sulfate, phenol, ethanol, caffeine, nicotine, and dimethyl sulfoxide were tested as model chemical stressors. Results obtained on a chip-based system were compared with static protocols performed in microtiter plates. This work provides evidence that FET analysis performed under microperfusion opens a brand new alternative for inexpensive automation in aquatic ecotoxicology.

International audience; Hospital wastewater (HWW) contain a large number of chemical pollutants such as disinfectants, surfactants, and pharmaceutical residues. A part of these pollutants is not eliminated by traditional urban wastewater treatment plants (WWTP), leading to a risk for the aquatic ecosystems receiving these effluents. In order to assess this risk, we formulated a specific methodology based on the ecotoxicological characterisation of the hospital wastewater using a battery of three chronic bioassays (Pseudokirchneriella subcapitata, Heterocypris incongruens and Brachionus calyciflorus). We used it for the posteriori risk assessment of a hospital recently built in south-east France, and we studied the evolution of this risk during two years. We also used it to assess the decrease of the ecotoxicological risk after treatment of the effluent in a specific line of the local WWTP. Lastly, we compared these results with the risk assessment made before the building of the hospital in the context of a priori risk assessment. The results obtained showed an important evolution of the risk overtime, according to the hospital activities and the river flows, and a real decrease of the risk after treatment in the dedicated line. They also showed that the a priori assessment of ecotoxicological risks, made previously, was overstated, mainly because of the application of the precautionary principle.

International audience; Tamoxifen, a drug used to treat cancer, is regularly found in hydrosystems at concentrations of several hundred ng L-1. To characterize its ecotoxicity, we implemented a battery of bioassays on organisms belonging to 3 different trophic levels: Pseudokirchneriella subcapitata, Chlorella vulgaris and Chlamydo- monas reinhardtii, for primary producers, Daphnia magna (immobilization, grazing and reproduction) for primary consumers, and Danio rerio for secondary consumers (embryotoxicity test). In view of the results obtained and the ecotoxicity values of tamoxifen available in the literature, we established a PNEC (Predictive No Effect Concentration) equal to 81 ng L-1 for continental water. This PNEC allowed us to cal- culate Risk Quotients (RQ) for 4 continental hydrosystems in 4 different countries in which measures of tamoxifen had already been performed on surface waters. In two of the situations studied, RQs were higher than 1, reaching a maximum of 2.6. These results show the need to deepen the characterization of ecotoxicological risks linked to the discharge of tamoxifen in surface waters. In addition, we propose applying this approach to other drug residues detected in the environment.