Anne Bassères

Influence of temperature, pH, oxygenation, water-type and substrate on biomarker responses in the freshwater clam Corbicula fluminea (Müller)

We investigated the influence of abiotic factors on responses of components of detoxification metabolism of phases I and II, indicators related to oxidative stress and propionylcholinesterase, proposed as biomarkers of pollution in Corbicula fluminea. Combined effects of temperature (10 and 20 degrees C), water-type (water purified by reverse-osmosis, lake water from the collection site and tap water) and habitat substrate (presence and absence of sand) were assessed in a five-day experiment. Additionally, clams were exposed to hypoxia or submitted to acidic, neutral or alkaline conditions at 20 degrees C, in lake water, without sand, for five days. Responses of biomarkers were generally higher when clams were placed in tap water, except for level of peroxidised lipids (PL). Catalase (CAT) and NADH-cytochrome c reductase (NADH-red.) activities increased whereas propionylcholinesterase (PChE) activity decreased in absence of sand. Decreasing temperature resulted in depressing PChE and NADPH-cytochrome c reductase (NADPH-red.) activities. Hypoxia induced an increase of CAT activity and glutathione S-transferase activity towards ethacrynic acid and a decrease of PL level. CAT and NADH-red. activities as well as PL level were reduced when clams were exposed to acidic and/or alkaline conditions. These results indicate that effects of abiotic factors should be taken into account in environmental studies.

Seasonal variations of pollution biomarkers in two populations of Corbicula fluminea (Müller)

Seasonal variations of pollution biomarkers have been studied in two populations of Corbicula fluminea (Müller) for 2 years. Clams were collected from the Sanguinet lake and the Dronne river (Southwest France), sites characterised by water of excellent and passable quality, respectively. Biological parameters studied were enzymatic activity (catalase, propionylcholinesterase, glutathione S-transferase, NADPH-cytochrome c reductase and NADH-cytochrome c reductase activities) and indicators of physiological (protein, carbohydrate, lipid and ash contents, fresh and dry tissue weights, condition factor) and reproductive (amount of oocytes present in gonads and clams incubating larvae in gills) status. Environmental parameters (e.g. water temperature, chlorophyll a concentration) were also monitored. Effects of site, sampling date and season (from spring to winter) were studied, as well as relationships between abiotic and biotic factors. Finally, an index of variability was calculated for each biomarker. Catalase, propionylcholinesterase, NADH-cytochrome c reductase and lipids experienced the most pronounced seasonal fluctuations, whereas glutathione S-transferase, NADPH-cytochrome c reductase and proteins suffered less pronounced ones. For some parameters, results differed depending on the site. Most seasonal changes were related to the reproductive cycle and/or water temperature. Implications of such variations in the use of pollution biomarkers in environmental surveys are discussed.

Potential biomarkers of trichloroethylene and toluene exposure in Corbicula fluminea.

Freshwater clams Corbicula fluminea were exposed in aquariums to four doses of trichloroethylene-TCE-(1.56 up to 100 mg/1) or toluene-TOL-(7.5 up to 60 mg/1) for 5 days. At the end of exposure, components of (de)toxification metabolism of phases I and II, parameters related to oxidative stress and propionylcholinesterase activity were assayed. Determination of TCE and TOL concentrations in water revealed an important evaporative loss during the experiment, characteristic of acute and occasional contaminations by such products occurring in the environment. Appropriate statistical methods such as ANOVA, Tukey test and discriminant analysis underlined the relevance of cytochromes P450 and P418, NADH-cytochrome c reductase, catalase, peroxided and peroxidizable lipids and net peroxidation as biomarkers of exposure to these solvents in C. fluminea. This experiment emphasised the importance of a multi-biomarker approach in environmental surveys and will be completed further by mesocosm studies.

Response of the bivalve Unio tumidus and freshwater communities in artificial streams for hazard assessment of methyl methacrylate

In artificial streams (pilot rivers) supplied with the river water of Gave de Pau (France), we studied the effects of methyl methacrylate (MMA) on the autochthonous bivalve Unio tumidus transferred into the streams and on natural freshwater communities colonizing the channels. Unio tumidus and freshwater communities were exposed to MMA for 15 and 30 d, respectively, at measured concentrations ranging from 0.6 to 122 mg/L. Biomarkers studied in the digestive gland and gills of U. tumidus comprised detoxication systems (namely, antioxidant enzyme activities and glutathione status) and lipid peroxidation as a marker of cytotoxicity. Biocoenotic indicators were used to evaluate effects on benthic invertebrates and diatoms. In bivalves, a decrease in antioxidant levels was found at the lowest concentrations tested (0.6 and 6 mg/L), whereas an increase in lipid peroxidation and mortality was registered at 30 mg/L after 15 d of exposure. Disturbances in freshwater communities occurred after 30 d at MMA concentrations of greater than 30 mg/L. Antioxidant responses in bivalves were recorded at the lowest MMA concentration tested, which was close to the predicted no-effect concentration (0.74 mg/L), and cytotoxicity was registered at a concentration corresponding to the 21-d no-observed-effect concentration for Daphnia magna. On the basis of the criteria studied, antioxidant biomarkers of bivalves appeared to be more sensitive than biocoenotic indicators to MMA.

Isotopic niche metrics as indicators of toxic stress in two freshwater snails

Descriptors of trophic niche and of food web structure and function have been suggested as integrative and sensitive endpoints of toxicant effects. In the present study, carbon and nitrogen stable isotope signatures were used to assess the effects of the dithiocarbamate fungicide thiram (35 and 170μg/L nominal concentrations) and of a petroleum distillate (0.01, 0.4, 2 and 20mg/L nominal loadings as Hydrocarbon Emulsion or Hydrocarbon Water Accommodated Fraction) on the trophic niche of two freshwater gastropods in artificial streams (Radix peregra) and ponds (Lymnaea stagnalis). Results were analyzed using classical univariate statistical methods and recently proposed uni- and multivariate metrics of the realized trophic niche of species. The trophic niche metrics were highly sensitive to both types of chemicals, but exposure resulted in different response patterns according to the nature of the tested compound. Thiram clearly affected gastropod trophic niche leading to a change in the food resources used and resulting in trophic niche expansion (i.e., increase of diversity of used resources, especially dead animals) or trophic niche contraction (i.e., decrease of diversity of used resources) across time. Both gastropod taxa exposed to hydrocarbons showed a clear trophic niche expansion. Trophic niche metrics therefore provide a promising way of investigating non-lethal effects of exposure to organic chemicals on aquatic invertebrates, and subsequent disturbances in food webs.

Secondary production of freshwater zooplankton communities exposed to a fungicide and to a petroleum distillate in outdoor pond mesocosms

Ecological risk assessment of chemicals in mesocosms requires measurement of a large number of parameters at the community level. Studies on invertebrate communities usually focus on taxonomic approaches, which only provide insights into taxonomic structure changes induced by chemicals. In the present study, abundance, biomass (B), theoretical production (P), and instantaneous P/B ratio were used as endpoints to assess the effects of the commercial form of the dithiocarbamate fungicide thiram (35 µg/L and 170 µg/L nominal concentrations) and of the hydrocarbon water accommodated fraction (HWAF) of a petroleum distillate (0.01 mg/L, 0.4 mg/L, 2 mg/L, and 20 mg/L loadings) on the zooplankton community in freshwater pond mesocosms. Endpoints were measured during a 4-wk treatment period (1 pulse/wk) followed by a 5-mo posttreatment period to evaluate zooplankton population recovery. The chlorophyll a concentration in water was significantly increased after treatment with HWAF, whereas it was not affected by thiram treatment. Zooplankton abundance-based analysis showed effects on a limited number of taxa, whereas other endpoints (mainly the P/B ratio) revealed that more taxa were impacted, with recovery depending on the chemical and concentration. Exposure to HWAF mainly had a negative impact on cladocerans, which resulted in top-down effects (between cladocerans and phytoplankton). Thiram negatively affected rotifers and copepods, suggesting more direct toxic effects. The results show that the use of secondary production as an endpoint provides a more comprehensive assessment of potential direct and indirect effects of chemicals on a community, and they also support evidence of alteration in functional processes.

Sensitivity ranking for freshwater invertebrates towards hydrocarbon contaminants

Hydrocarbons have an utmost economical importance but may also cause substantial ecological impacts due to accidents or inadequate transportation and use. Currently, freshwater biomonitoring methods lack an indicator that can unequivocally reflect the impacts caused by hydrocarbons while being independent from effects of other stressors. The aim of the present study was to develop a sensitivity ranking for freshwater invertebrates towards hydrocarbon contaminants, which can be used in hydrocarbon-specific bioindicators. We employed the Relative Sensitivity method and developed the sensitivity ranking Shydrocarbons based on literature ecotoxicological data supplemented with rapid and mesocosm test results. A first validation of the sensitivity ranking based on an earlier field study has been conducted and revealed the Shydrocarbons ranking to be promising for application in sensitivity based indicators. Thus, the first results indicate that the ranking can serve as the core component of future hydrocarbon-specific and sensitivity trait based bioindicators.