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    Laurent Lagadic

    Long-term effects of endocrine disruptors (EDs) on aquatic invertebrates remain difficult to assess, mainly due to the lack of appropriate sensitive toxicity test methods and relevant data analysis procedures. This study aimed at... more
    Long-term effects of endocrine disruptors (EDs) on aquatic invertebrates remain difficult to assess, mainly due to the lack of appropriate sensitive toxicity test methods and relevant data analysis procedures. This study aimed at identifying windows of sensitivity to EDs along the life-cycle of the freshwater snail Lymnaea stagnalis, a candidate species for the development of forthcoming test guidelines. Juveniles, sub-adults,
    ABSTRACT
    To optimize their efficacy, some insecticides used for mosquito control are introduced into aquatic ecosystems where mosquito larvae develop (marshes, ponds, sanitation devices) and cannot escape from the treated water. However, this... more
    To optimize their efficacy, some insecticides used for mosquito control are introduced into aquatic ecosystems where mosquito larvae develop (marshes, ponds, sanitation devices) and cannot escape from the treated water. However, this raises the question of possible effects of mosquito larvicides on non-target aquatic species. Bacillus thuringiensis var. israelensis (Bti), which is well-known for its selectivity for Nematocera dipterans, is widely used for mosquito control all over the world. Spinosad, a mixture of spinosyns A and D known as fermentation products of a soil actinomycete (Saccharopolyspora spinosa), is a biological neurotoxic insecticide with a broader action spectrum. It is a candidate larvicide for mosquito control, but some studies showed that it may be toxic to beneficial or non-target species, including non-biting midges. The present study was therefore undertaken to assess the impact of Bti and spinosad on natural populations of Polypedilum nubifer (Skuse) and Ta...
    The sensitivity of diatom taxonomy and trait-based endpoints to chemicals has been poorly used so far in Environmental Risk Assessment. In this study, diatom assemblages in outdoor flow-through mesocosms were exposed to thiram (35 and... more
    The sensitivity of diatom taxonomy and trait-based endpoints to chemicals has been poorly used so far in Environmental Risk Assessment. In this study, diatom assemblages in outdoor flow-through mesocosms were exposed to thiram (35 and 170 μg/L), and a hydrocarbon emulsion (HE; 0.01, 0.4, 2 and 20 mg/L). The effects of exposure were assessed for 12 weeks, including 9 weeks post-treatment, using taxonomic structure and diversity, bioindication indices, biological traits, functional diversity indices, indicator classes and ecological guilds. For both chemicals, diversity increased after the treatment period, and responses of ecological traits were roughly identical with an abundance increase of motile taxa tolerant to organic pollution and decrease of low profile taxa. Bioindication indices were not affected. Traits provided a complementary approach to biomass measurements and taxonomic descriptors, leading to a more comprehensive overview of ecological changes due to organic chemicals...
    Micro- and mesocosms are frequently required in regulatory procedures of aquatic risk assessment for pesticides. However, many questions are still a matter of debate with regard to the use of these systems for environmental risk... more
    Micro- and mesocosms are frequently required in regulatory procedures of aquatic risk assessment for pesticides. However, many questions are still a matter of debate with regard to the use of these systems for environmental risk assessment, especially considering the inter-system variability of the measured parameters and its consequences on experimental design and data analysis. In this paper, variability of physico-chemical and biological parameters measured during two long-term experiments (8 to 9 months) in uncontaminated outdoor freshwater lentic mesocosms (8 m3) is analysed. Consequences on the design of ecotoxicity tests in mesocosms and on data analysis are also addressed. Water temperature, pH, dissolved oxygen concentration and concentration of suspended solids exhibited a very low variability whereas nutrient concentrations displayed elevated levels of variability. Among biological parameters, those measured at the individual level were less variable than those measured a...
    The OECD test guideline development program has been extended in 2011 to establish a partial life-cycle protocol for assessing the reproductive toxicity of chemicals to several mollusk species, including the great pond snail Lymnaea... more
    The OECD test guideline development program has been extended in 2011 to establish a partial life-cycle protocol for assessing the reproductive toxicity of chemicals to several mollusk species, including the great pond snail Lymnaea stagnalis. In this paper, we summarize the standard draft protocol for a reproduction test with this species, and present inter-comparison results obtained in a 56-day prevalidation ring-test using this protocol. Seven European laboratories performed semi-static tests with cultured snails of the strain Renilys® exposed to nominal concentrations of cadmium chloride (from 53 to 608μgCdL(-1)). Cd concentrations in test solutions were analytically determined to confirm accuracy in the metal exposure concentrations in all laboratories. Physico-chemical and biological validity criteria (namely dissolved oxygen content >60% ASV, water temperature 20±1°C, control snail survival >80% and control snail fecundity >8 egg-masses per snail over the test period) were met in all laboratories which consistently demonstrated the reproductive toxicity of Cd in snails using the proposed draft protocol. Effect concentrations for fecundity after 56days were reproducible between laboratories (68<EC50-56d<124μgL(-1)) and were consistent with literature data. EC50-56d and EC10-56d values were comprised within a factor of 1.8 and 3.6, respectively, which is in the range of acceptable variation defined for reference chemicals in OECD test guidelines for invertebrates. The inter-laboratory reproducibility coefficient of variation (CV) for the Cd LC50-56d values was 8.19%. The inter-laboratory comparison of fecundity within the controls gave a CV of 29.12%, while exposure to Cd gave a CV of 25.49% based on the EC50-56d values. The OECD has acknowledged the success of this prevalidation exercise and a validation ring-test involving 14 laboratories in Europe, North- and South-America is currently being implemented using four chemicals (Cd, prochloraz, trenbolone and tributyltin).
    This study examined the response of 7-ethoxyresorufine-O-deethylase, glutathione-S-transferase, glutathione peroxidase, glutathione content, level of thiobarbituric acid reactive compounds and circulating vitellogenin, in three-spined... more
    This study examined the response of 7-ethoxyresorufine-O-deethylase, glutathione-S-transferase, glutathione peroxidase, glutathione content, level of thiobarbituric acid reactive compounds and circulating vitellogenin, in three-spined sticklebacks after 21 days of exposure to Diquat herbicide, commercial nonylphenol polyethoxylate adjuvant and mixture between Diquat and adjuvant. The results showed that adjuvant exerted more important oxidative effects than Diquat and that mixture effects were unlike to single additivity. This study argues for ecotoxicological risk assessment of adjuvants and mixtures of adjuvants and pesticides.
    The aim of this review is to examine how the choice of test species and study design employed in the use of in situ approaches in ecological risk assessment can maximize the ecological relevance of data. We provide a framework to define... more
    The aim of this review is to examine how the choice of test species and study design employed in the use of in situ approaches in ecological risk assessment can maximize the ecological relevance of data. We provide a framework to define and assess ecological relevance that permits study designs to remain focused on the ecological question being addressed. This framework makes explicit the linkages between effects at lower levels of biological organization and higher-order ecological effects at the population, community, and ecosystem levels. The usefulness of this framework is illustrated by reference to specific examples from aquatic ecotoxicology. The use of models as both interpretive and predictive tools is discussed, with suggestions of appropriate methods for different protection goals.
    The ecotoxicological impact of nitrate-induced photodegradation products of diuron and chlorotoluron was studied through monospecific biotests conducted in conjunction with experiments in outdoor aquatic mesocosms. Organisms representing... more
    The ecotoxicological impact of nitrate-induced photodegradation products of diuron and chlorotoluron was studied through monospecific biotests conducted in conjunction with experiments in outdoor aquatic mesocosms. Organisms representing three trophic levels were used: two heterotrophic microorganisms, the luminescent bacterium Vibrio fischeri and the ciliated protozoa Tetrahymena pyriformis, and one metazoa, the gastropod Lymnaea stagnalis. Among the variety of the phenylurea photoproducts, the N-formylated ones appeared clearly more toxic than the parent compounds towards the microorganisms, whereas the nitroderivatives showed a similar toxicity. Using photodegraded solutions of diuron, toxicity was maintained or even increased during disappearance of the initial herbicide, demonstrating that some of the photoproducts may have an impact additively or in synergy. Enzymatic biomarker assays performed on Lymnaea stagnalis exposed under monospecific conditions showed significant effects, due to the combination of nitrate with the pesticide and its photoproducts. A positive impact on snail fecundity was observed with chlorotoluron both under monospecific laboratory and integrated mesocosm conditions. Oviposition stimulation took place when first- and second-generation photoproducts were predominant.
    The present study was undertaken to assess the impact of a candidate mosquito larvicide, spinosad (8, 17 and 33 microg L(-1)) on a field population of Daphnia magna under natural variations of water temperature and salinity, using Bti... more
    The present study was undertaken to assess the impact of a candidate mosquito larvicide, spinosad (8, 17 and 33 microg L(-1)) on a field population of Daphnia magna under natural variations of water temperature and salinity, using Bti (0.16 and 0.50 microL L(-1)) as the reference larvicide. Microcosms (125 L) were placed in a shallow temporary marsh where D. magna was naturally present. The peak of salinity observed during the 21-day observation period may have been partly responsible for the decrease of daphnid population density in all the microcosms. It is also probably responsible for the absence of recovery in the microcosms treated with spinosad which caused a sharp decrease of D. magna abundance within the first two days following treatment whereas Bti had no effect. These results suggest that it may be difficult for a field population of daphnids to cope simultaneously with natural (water salinity and temperature) and anthropogenic (larvicides) stressors.
    The influence of tank-mix adjuvants on pesticide toxicity remains largely unknown. Agral 90, a nonylphenol polyethoxylated tank-mix adjuvant, has been used with diquat (bipyridylium herbicide) and fomesafen (diphenyl-ether herbicide) in... more
    The influence of tank-mix adjuvants on pesticide toxicity remains largely unknown. Agral 90, a nonylphenol polyethoxylated tank-mix adjuvant, has been used with diquat (bipyridylium herbicide) and fomesafen (diphenyl-ether herbicide) in aquatic indoor microcosms in order to compare the toxicity of the single compounds and of binary herbicide-adjuvant mixtures to Lemna minor. Twenty-four microcosms were used and treatments were performed with substances alone or with herbicide-adjuvant binary mixtures, at two concentrations levels (44.4 and 222.2 microg/L for the herbicides, and 100 and 500 microg/L for Agral 90). Toxicity was assessed weekly for 1 month through growth measurements, as inferred from the relative frond number (RFN) and relative frond area (RFA). Concentrations of diquat and fomesafen in water and sediments were measured weekly. The herbicides showed very different behaviour in microcosms, with a rapid disappearance of diquat from the aqueous phase whereas fomesafen levels remained almost constant over time. Diquat strongly inhibited the growth of L. minor whereas fomesafen had no effect on plant growth. Presence of the adjuvant only slightly reduced the effect of the lowest concentration of diquat, probably as a result of dispersion of the herbicide at the water surface. It is concluded that tank-mix adjuvant designed to improve herbicide efficiency in the terrestrial environment did not have any effect on aquatic plants when applied to the aquatic environment.
    Bacillus thuringiensis var. israelensis (Bti) is commonly used for selective control of larval populations of mosquitoes in coastal wetlands. A two year-study was implemented to investigate whether repeated treatments with Bti applied... more
    Bacillus thuringiensis var. israelensis (Bti) is commonly used for selective control of larval populations of mosquitoes in coastal wetlands. A two year-study was implemented to investigate whether repeated treatments with Bti applied either as a liquid (VectoBac® 12AS) or a water-dispersible granule (VectoBac® WG) formulation may affect the abundance and diversity of non-target aquatic invertebrates in saltmarsh pools. Taxonomic composition of the invertebrate communities was typical of brackishwater intermittent ecosystems, with a dominance of annelids, crustaceans and nematocerans. Conditions were contrasted between the two years of the survey, both in terms of annual cumulative rainfall and rainfall distribution throughout the year. As a consequence, the hydroperiod and some other environmental characteristics associated with pool drying played a major role in the dynamics of the invertebrate community. In summer 2006, pool drying reduced the abundance of the polychaete worm Nereis diversicolor, of the amphipod crustacean Corophium volutator and of chironomid larvae. These taxa were able to recolonize rapidly the pools after flooding in September 2006. In 2007, rainfall was more regularly distributed across the year, and the pools did not get dry. Hydrozoans, Chironomini and Orthocladiinae larvae, and oligochaetes were more abundant in treated than in control pools, especially in VectoBac® WG-treated pools. No adverse effects of the treatments were shown on the abundance of N. diversicolor, C. volutator and midge larvae, suggesting that the availability of these food sources for birds was not negatively affected by Bti applications. It is concluded that, as currently performed in Western France coastal wetlands, land-based treatments of saltmarsh pools for larval mosquito control with Bti, used either as VectoBac® 12AS or VectoBac® WG, did not adversely impact non-target aquatic invertebrate communities.
    Chitobiase is involved in exoskeleton degradation and recycling during the moulting process in arthropods. In aquatic species, the moulting fluid is released into the aqueous environment, and chitobiase activity present therein can be... more
    Chitobiase is involved in exoskeleton degradation and recycling during the moulting process in arthropods. In aquatic species, the moulting fluid is released into the aqueous environment, and chitobiase activity present therein can be used to follow the dynamics of arthropod populations. Here, chitobiase activity was used for monitoring the impact of mosquito candidate larvicides on Daphnia pulex and Daphnia magna under laboratory conditions. Both species were exposed to spinosad (2, 4, 8 μg L(-1)) and diflubenzuron (0.2, 0.4, 0.8 μg L(-1)) for 14 days. Bacillus thuringiensis var. israelensis (Bti; 0.25, 0.5, 1 μL L(-1)) was used as the reference larvicide. Chitobiase activity, adult survival, individual growth and fecundity, expressed as the number of neonates produced, were measured every 2 days. Average Exposure Concentrations of spinosad were ten-fold lower than the nominal concentrations, whereas only a slight deviation was observed for diflubenzuron. In contrast to Bti, spinosad and diflubenzuron significantly affected both species in terms of adult survival, and production of neonates. As compared to D. pulex, D. magna was more severely affected by diflubenzuron, at low and medium concentrations, with reduced adult growth and much lower chitobiase activity. Chitobiase activity was positively correlated with the individual body length, number of neonates produced between two consecutive observation dates, and number of females and neonates. In addition, the significant positive correlations between chitobiase activity measured on the last sampling date before the first emission of neonates and the cumulative number of neonates produced during the whole observation period strongly support the potential of the activity of this chitinolytic enzyme as a proxy for assessing the dynamics of arthropod populations exposed to larvicides used for mosquito control.
    A series of four papers, commissioned by the European Science Foundation, are presented on the state-of-the-art of the use of biomarkers in environmental assessment. These papers are phylogenetically based and cover invertebrates,... more
    A series of four papers, commissioned by the European Science Foundation, are presented on the state-of-the-art of the use of biomarkers in environmental assessment. These papers are phylogenetically based and cover invertebrates, vertebrates, plants and invertebrate populations and communities.
    When spiked in sediments, copper is known to reduce growth of Chironomus riparius larvae and the production of eggs by adult females. The aim of this work was to better understand the origin of these phenomena by studying the effects of... more
    When spiked in sediments, copper is known to reduce growth of Chironomus riparius larvae and the production of eggs by adult females. The aim of this work was to better understand the origin of these phenomena by studying the effects of copper using developmental and energetic biomarkers, such as changes in larval weight and age and changes in the levels of sugars and lipids. Four-day-old C. riparius larvae were exposed to nominal concentrations of copper of 0, 6.5, 12.5, 25 and 50 mg/kg of dry sediment (silica) in 0.6 l beakers. They were fed ad libitum and exposures were stopped at 7 and 9 days after the beginning of the tests. The larvae were weighed, sexed and aged. For each sex, the larvae belonging to the phases the most frequently found in the beakers were selected for dissection and measurement of energy reserves. The increase in the concentration of copper resulted in an increasing delay in larval growth in both sexes. Desynchronized development was observed, as shown by the increase in the number of individuals that remained in the third instar or early phases of the fourth instar, as well as by a reduction in age of males. Concerning energy reserves, the levels of sugars (glycogen, trehalose and glucose) in the dissected larvae remained almost constant among levels of exposure. In contrast, at the highest copper concentration (50 mg/kg), triglyceride levels suffered a slight reduction whereas the level of free glycerol significantly increased. It is concluded that selection of C. riparius larvae for both sex and age improves the relevance of some energy-yielding substrates as indicators of adverse physiological effects of copper.