The release of polycyclic aromatic hydrocarbons (PAHs) into the environment has increased very substantially over the last decades leading to high concentrations in sediments of contaminated areas. To evaluate the consequences of long-term chronic exposure to PAHs, zebrafish were exposed, from their first meal at 5 days post fertilisation until they became reproducing adults, to diets spiked with three PAH fractions at three environmentally relevant concentrations with the medium concentration being in the range of 4.6–6.7 μg g−1 for total quantified PAHs including the 16 US-EPA indicator PAHs and alkylated derivatives. The fractions used were representative of PAHs of pyrolytic (PY) origin or of two different oils of differing compositions, a heavy fuel (HO) and a light crude oil (LO). Fish growth was inhibited by all PAH fractions and the effects were sex specific: as determined with 9-month-old adults, exposure to the highest PY inhibited growth of females; exposure to the highest HO and LO inhibited growth of males; also, the highest HO dramatically reduced survival. Morphological analysis indicated a disruption of jaw growth in larvae and malformations in adults. Intestinal and pancreatic enzyme activities were abnormal in 2-month-old exposed fish. These effects may contribute to poor growth. Finally, our results indicate that PAH mixtures of different compositions, representative of situations encountered in the wild, can promote lethal and sublethal effects which are likely to be detrimental for fish recruitment.

The release of polycyclic aromatic hydrocarbons (PAHs) into the environment has increased very substantially over the last decades leading to high concentrations in sediments of contaminated areas. To evaluate the consequences of long-term chronic exposure to PAHs, zebrafish were exposed, from their first meal at 5 days post fertilisation until they became reproducing adults, to diets spiked with three PAH fractions at three environmentally relevant concentrations with the medium concentration being in the range of 4.6–6.7 μg g−1 for total quantified PAHs including the 16 US-EPA indicator PAHs and alkylated derivatives. The fractions used were representative of PAHs of pyrolytic (PY) origin or of two different oils of differing compositions, a heavy fuel (HO) and a light crude oil (LO). Fish growth was inhibited by all PAH fractions and the effects were sex specific: as determined with 9-month-old adults, exposure to the highest PY inhibited growth of females; exposure to the highest HO and LO inhibited growth of males; also, the highest HO dramatically reduced survival. Morphological analysis indicated a disruption of jaw growth in larvae and malformations in adults. Intestinal and pancreatic enzyme activities were abnormal in 2-month-old exposed fish. These effects may contribute to poor growth. Finally, our results indicate that PAH mixtures of different compositions, representative of situations encountered in the wild, can promote lethal and sublethal effects which are likely to be detrimental for fish recruitment.

The release of polycyclic aromatic hydrocarbons (PAHs) into the environment has increased very substantially over the last decades leading to high concentrations in sediments of contaminated areas. To evaluate the consequences of long-term chronic exposure to PAHs, zebrafish were exposed, from their first meal at 5 days post fertilisation until they became reproducing adults, to diets spiked with three PAH fractions at three environmentally relevant concentrations with the medium concentration being in the range of 4.6–6.7 μg g⁻¹for total quantified PAHs including the 16 US-EPA indicator PAHs and alkylated derivatives. The fractions used were representative of PAHs of pyrolytic (PY) origin or of two different oils of differing compositions, a heavy fuel (HO) and a light crude oil (LO). Fish growth was inhibited by all PAH fractions and the effects were sex specific: as determined with 9-month-old adults, exposure to the highest PY inhibited growth of females; exposure to the highest HO and LO inhibited growth of males; also, the highest HO dramatically reduced survival. Morphological analysis indicated a disruption of jaw growth in larvae and malformations in adults. Intestinal and pancreatic enzyme activities were abnormal in 2-month-old exposed fish. These effects may contribute to poor growth. Finally, our results indicate that PAH mixtures of different compositions, representative of situations encountered in the wild, can promote lethal and sublethal effects which are likely to be detrimental for fish recruitment.

International audience | Chlordecone is an organochlorine insecticide that has been widely used to control banana weevil in the French West Indies. As a result of this intense use, up to 20,000 ha are contaminated by this insecticide in the French West Indies, and this causes environmental damage and health problems. A scenario of exposure was drawn by French authorities, based on land usage records. Many efforts have been made to monitor the occurrence of chlordecone and its main metabolites using different analytical methods, including GC, GC/MS, LC/MS, and NIRS. Although these different methods allow for the detection and quantification of chlordecone from soils, none of them estimate the bottleneck caused by extraction of this organochlorine from soils with high adsorption ability. In this study, we used C-13(10)-chlordecone as a tracer to estimate chlordecone extraction yield and to quantify chlordecone in soil extracts based on the C-13/C-12 isotope dilution. We report the optimization of C-13(10)-chlordecone extraction from an Andosol. The method was found to be linear from 0.118 to 43 mg kg(-1) in the Andosol, with an instrumental detection limit estimated at 8.84 mu g kg(-1). This method showed that chlordecone ranged from 35.4 down to 0.18 mg kg(-1) in Andosol, Nitisol, Ferralsol, and Fluvisol soil types. Traces of the metabolite beta-monohydrochlordecone were detected in the Andosol, Nitisol, and Ferralsol soil samples. This last result indicates that this method could be useful to monitor the fate of chlordecone in soils of the French West Indies.

Chlordecone is an organochlorine insecticide that has been widely used to control banana weevil in the French West Indies. As a result of this intense use, up to 20,000 ha are contaminated by this insecticide in the French West Indies, and this causes environmental damage and health problems. A scenario of exposure was drawn by French authorities, based on land usage records. Many efforts have been made to monitor the occurrence of chlordecone and its main metabolites using different analytical methods, including GC, GC/MS, LC/MS, and NIRS. Although these different methods allow for the detection and quantification of chlordecone from soils, none of them estimate the bottleneck caused by extraction of this organochlorine from soils with high adsorption ability. In this study, we used¹³C₁₀-chlordecone as a tracer to estimate chlordecone extraction yield and to quantify chlordecone in soil extracts based on the¹³C/¹²C isotope dilution. We report the optimization of¹³C₁₀-chlordecone extraction from an Andosol. The method was found to be linear from 0.118 to 43 mg kg⁻¹in the Andosol, with an instrumental detection limit estimated at 8.84 μg kg⁻¹. This method showed that chlordecone ranged from 35.4 down to 0.18 mg kg⁻¹in Andosol, Nitisol, Ferralsol, and Fluvisol soil types. Traces of the metabolite β-monohydrochlordecone were detected in the Andosol, Nitisol, and Ferralsol soil samples. This last result indicates that this method could be useful to monitor the fate of chlordecone in soils of the French West Indies.

International audience | In the last 10 years, behavior assessment has been developed as an indicator of neurotoxicity and an integrated indicator of physiological disruption. Polycyclic aromatic hydrocarbon (PAH) release into the environment has increased in recent decades resulting in high concentrations of these compounds in the sediment of contaminated areas. We evaluated the behavioral consequences of long-term chronic exposure to PAHs, by exposing zebrafish to diets spiked with three PAH fractions at environmentally relevant concentrations. Fish were exposed to these chemicals from their first meal (5 days postfertilization) until they became reproducing adults (at 6 months old). The fractions used were representative of PAHs of pyrolytic (PY) origin and of two oils differing in composition (a heavy fuel oil (HO) and a light crude oil (LO)). Several tests were carried out to evaluate circadian spontaneous swimming activity, responses to a challenge (photomotor response), exploratory tendencies, and anxiety levels. We found that dietary PAH exposure was associated with greater mobility, lower levels of exploratory activity, and higher levels of anxiety, particularly in fish exposed to the HO fraction and, to a lesser extent, the LO fraction. Finally, our results indicate that PAH mixtures of different compositions, representative of situations encountered in the wild, can induce behavioral disruptions resulting in poorer fish performance.

In the last 10 years, behavior assessment has been developed as an indicator of neurotoxicity and an integrated indicator of physiological disruption. Polycyclic aromatic hydrocarbon (PAH) release into the environment has increased in recent decades resulting in high concentrations of these compounds in the sediment of contaminated areas. We evaluated the behavioral consequences of long-term chronic exposure to PAHs, by exposing zebrafish to diets spiked with three PAH fractions at environmentally relevant concentrations. Fish were exposed to these chemicals from their first meal (5 days postfertilization) until they became reproducing adults (at 6 months old). The fractions used were representative of PAHs of pyrolytic (PY) origin and of two oils differing in composition (a heavy fuel oil (HO) and a light crude oil (LO)). Several tests were carried out to evaluate circadian spontaneous swimming activity, responses to a challenge (photomotor response), exploratory tendencies, and anxiety levels. We found that dietary PAH exposure was associated with greater mobility, lower levels of exploratory activity, and higher levels of anxiety, particularly in fish exposed to the HO fraction and, to a lesser extent, the LO fraction. Finally, our results indicate that PAH mixtures of different compositions, representative of situations encountered in the wild, can induce behavioral disruptions resulting in poorer fish performance.

International audience | We expose here a detailed spatially explicit model of aphid population dynamics at the scale of a whole country (Metropolitan France). It is based on convection-diffusionreaction equations, driven by abiotic and biotic factors. The target species is the grain aphid, Sitobion avenae F., considering both its winged and apterousmorphs. In this preliminary work, simulations for year 2004 (an outbreak case) produced realistic aphid densities, and showed that both spatial and temporal S. avenae population dynamics can be represented as an irregular wave of population peak densities from southwest to northeast of the country, driven by gradients or differences in temperature, wheat phenology, and wheat surfaces. This wave pattern fits well to our knowledge of S. avenae phenology. The effects of three insecticide spray regimes were simulated in five different sites and showed that insecticide sprays were ineffective in terms of yield increase after wheat flowering. After suitable validation, which will require some further years of observations, the model will be used to forecast aphid densities in real time at any date or growth stage of the crop anywhere in the country. It will be the backbone of a decision support system, forecasting yield losses at the level of a field. The model intends then to complete the punctual forecasting provided by older models by a comprehensive spatial view on a large area and leads to the diminution of insecticide sprayings in wheat crops.

We expose here a detailed spatially explicit model of aphid population dynamics at the scale of a whole country (Metropolitan France). It is based on convection–diffusion-reaction equations, driven by abiotic and biotic factors. The target species is the grain aphid, Sitobion avenae F., considering both its winged and apterous morphs. In this preliminary work, simulations for year 2004 (an outbreak case) produced realistic aphid densities, and showed that both spatial and temporal S. avenae population dynamics can be represented as an irregular wave of population peak densities from southwest to northeast of the country, driven by gradients or differences in temperature, wheat phenology, and wheat surfaces. This wave pattern fits well to our knowledge of S. avenae phenology. The effects of three insecticide spray regimes were simulated in five different sites and showed that insecticide sprays were ineffective in terms of yield increase after wheat flowering. After suitable validation, which will require some further years of observations, the model will be used to forecast aphid densities in real time at any date or growth stage of the crop anywhere in the country. It will be the backbone of a decision support system, forecasting yield losses at the level of a field. The model intends then to complete the punctual forecasting provided by older models by a comprehensive spatial view on a large area and leads to the diminution of insecticide sprayings in wheat crops.

A new gravel-contact assay using rainbow trout, Oncorhynchus mykiss, embryos was developed to assess the toxicity of polycyclic aromatic hydrocarbons (PAHs) and other hydrophobic compounds. Environmentally realistic exposure conditions were mimicked with a direct exposure of eyed rainbow trout embryos incubated onto chemical-spiked gravels until hatching at 10 °C. Several endpoints were recorded including survival, hatching delay, hatching success, biometry, developmental abnormalities, and DNA damage (comet and micronucleus assays). This bioassay was firstly tested with two model PAHs, fluoranthene and benzo[a]pyrene. Then, the method was applied to compare the toxicity of three PAH complex mixtures characterized by different PAH compositions: a pyrolytic extract from a PAH-contaminated sediment (Seine estuary, France) and two petrogenic extracts from Arabian Light and Erika oils, at two environmental concentrations, 3 and 10 μg g−1 sum of PAHs. The degree and spectrum of toxicity were different according to the extract considered. Acute effects including embryo mortality and decreased hatching success were observed only for Erika oil extract. Arabian Light and pyrolytic extracts induced mainly sublethal effects including reduced larvae size and hemorrhages. Arabian Light and Erika extracts both induced repairable DNA damage as revealed by the comet assay versus the micronucleus assay. The concentration and proportion of methylphenanthrenes and methylanthracenes appeared to drive the toxicity of the three PAH fractions tested, featuring a toxic gradient as follows: pyrolytic < Arabian Light < Erika. The minimal concentration causing developmental defects was as low as 0.7 μg g−1 sum of PAHs, indicating the high sensitivity of the assay and validating its use for toxicity assessment of particle-bound pollutants.

A new gravel-contact assay using rainbow trout, Oncorhynchus mykiss, embryos was developed to assess the toxicity of polycyclic aromatic hydrocarbons (PAHs) and other hydrophobic compounds. Environmentally realistic exposure conditions were mimicked with a direct exposure of eyed rainbow trout embryos incubated onto chemical-spiked gravels until hatching at 10 °C. Several endpoints were recorded including survival, hatching delay, hatching success, biometry, developmental abnormalities, and DNA damage (comet and micronucleus assays). This bioassay was firstly tested with two model PAHs, fluoranthene and benzo[a]pyrene. Then, the method was applied to compare the toxicity of three PAH complex mixtures characterized by different PAH compositions: a pyrolytic extract from a PAH-contaminated sediment (Seine estuary, France) and two petrogenic extracts from Arabian Light and Erika oils, at two environmental concentrations, 3 and 10 μg g−1 sum of PAHs. The degree and spectrum of toxicity were different according to the extract considered. Acute effects including embryo mortality and decreased hatching success were observed only for Erika oil extract. Arabian Light and pyrolytic extracts induced mainly sublethal effects including reduced larvae size and hemorrhages. Arabian Light and Erika extracts both induced repairable DNA damage as revealed by the comet assay versus the micronucleus assay. The concentration and proportion of methylphenanthrenes and methylanthracenes appeared to drive the toxicity of the three PAH fractions tested, featuring a toxic gradient as follows: pyrolytic < Arabian Light < Erika. The minimal concentration causing developmental defects was as low as 0.7 μg g−1 sum of PAHs, indicating the high sensitivity of the assay and validating its use for toxicity assessment of particle-bound pollutants.

A multibiomarker approach was developed to evaluate the juvenile European flounder responses to a complex mixture of 9 polycyclic aromatic hydrocarbons (PAHs) and 12 polychlorinated biphenyls (PCBs). Exposure was performed through contaminated food pellets displaying: (1) PAH and PCB levels similar to those detected in the heavily polluted Seine estuary, respectively in sediments and in flatfish and (2) ten times these concentrations. Several biomarkers of the immune system (e.g., lysozyme concentration and gene expression of complement component C3 and TNF-receptor), DNA damage (e.g., Comet assay), energetic metabolism (e.g., activity of cytochrome C oxidase), detoxification process (e.g., cytochrome P450 1A1 expression level: CYP1A1; betaine homocysteine methyl transferase expression level: BHMT) were investigated after 14 and 29 days of contamination, followed by a 14-days recovery period. After 29 days of contamination, the detoxification activity (CYP1A1 expression level) was positively correlated with DNA damages; the increase of the BHMT expression level could also be related to the detoxification process. Furthermore, after the recovery period, some biomarkers were still upregulated (i.e., CYP1A1 and BHMT expression levels). The immune system was significantly modulated by the chemical stress at the two concentration levels, and the lysozyme appeared to be the most sensitive marker of the mixture impact.

A multibiomarker approach was developed to evaluate the juvenile European flounder responses to a complex mixture of 9 polycyclic aromatic hydrocarbons (PAHs) and 12 polychlorinated biphenyls (PCBs). Exposure was performed through contaminated food pellets displaying: (1) PAH and PCB levels similar to those detected in the heavily polluted Seine estuary, respectively in sediments and in flatfish and (2) ten times these concentrations. Several biomarkers of the immune system (e.g., lysozyme concentration and gene expression of complement component C3 and TNF-receptor), DNA damage (e.g., Comet assay), energetic metabolism (e.g., activity of cytochrome C oxidase), detoxification process (e.g., cytochrome P450 1A1 expression level: CYP1A1; betaine homocysteine methyl transferase expression level: BHMT) were investigated after 14 and 29 days of contamination, followed by a 14-days recovery period. After 29 days of contamination, the detoxification activity (CYP1A1 expression level) was positively correlated with DNA damages; the increase of the BHMT expression level could also be related to the detoxification process. Furthermore, after the recovery period, some biomarkers were still upregulated (i.e., CYP1A1 and BHMT expression levels). The immune system was significantly modulated by the chemical stress at the two concentration levels, and the lysozyme appeared to be the most sensitive marker of the mixture impact.

Pesticides are frequently detected in estuaries among the pollutants found in estuarine and coastal areas and may have major ecological consequences. They could endanger organism growth, reproduction or survival. In the context of high mortality outbreaks affecting Pacific oysters, Crassostrea gigas, in France since 2008, it appears of importance to determine the putative effects of pesticides on oyster susceptibility to infectious agents. Massive mortality outbreaks reported in this species, mainly in spring and summer, may suggest an important role played by the seasonal use of pesticides and freshwater input in estuarine areas where oyster farms are frequently located. To understand the impact of some pesticides detected in French waters, their effects on Pacific oyster hemocytes were studied through short-term in vitro experiments. Bivalve immunity is mainly supported by hemocytes eliminating pathogens by phagocytosis and producing compounds including lysosomal enzymes and antimicrobial molecules. In this study, oyster hemocytes were incubated with a mixture of 14 pesticides and metaldehyde alone, a molecule used to eliminate land mollusks. Hemocyte parameters including dead/alive cells, nonspecific esterase activities, intracytoplasmic calcium, lysosome number and activity and phagocytosis were monitored by flow cytometry. No significant effect of pesticides tested at different concentrations was reported on oyster hemocytes maintained in vitro for short-term periods in the present study. It could be assumed that these oyster cells were resistant to pesticide exposure in tested conditions and developing in vivo assays appears as necessary to better understand the effects of polluants on immune system in mollucks.

Pesticides are frequently detected in estuaries among the pollutants found in estuarine and coastal areas and may have major ecological consequences. They could endanger organism growth, reproduction, or survival. In the context of high-mortality outbreaks affecting Pacific oysters, Crassostrea gigas, in France since 2008, it appears of importance to determine the putative effects of pesticides on oyster susceptibility to infectious agents. Massive mortality outbreaks reported in this species, mainly in spring and summer, may suggest an important role played by the seasonal use of pesticides and freshwater input in estuarine areas where oyster farms are frequently located. To understand the impact of some pesticides detected in French waters, their effects on Pacific oyster hemocytes were studied through short-term in vitro experiments. Bivalve immunity is mainly supported by hemocytes eliminating pathogens by phagocytosis and producing compounds including lysosomal enzymes and antimicrobial molecules. In this study, oyster hemocytes were incubated with a mixture of 14 pesticides and metaldehyde alone, a molecule used to eliminate land mollusks. Hemocyte parameters including dead/alive cells, nonspecific esterase activities, intracytoplasmic calcium, lysosome number and activity, and phagocytosis were monitored by flow cytometry. No significant effect of pesticides tested at different concentrations was reported on oyster hemocytes maintained in vitro for short-term periods in the present study. It could be assumed that these oyster cells were resistant to pesticide exposure in tested conditions and developing in vivo assays appears as necessary to better understand the effects of pollutants on immune system in mollusks.

Most persistent organic pollutants, due to their hydrophobic properties, accumulate in aquatic sediments and represent a high risk for sediment quality. To assess the toxicity of hydrophobic pollutants, a novel approach was recently proposed as an alternative to replace, refine and reduce animal experimentation: the medaka embryo–larval sediment contact assay (MELAc). This assay is performed with Japanese medaka embryos incubated on a natural sediment spiked with the compound being tested. With the aim of improving this assay, our study developed a reference exposure protocol with an artificial sediment specifically designed to limit natural sediment composition uncertainties and preparation variability. The optimum composition of the new artificial sediment was tested using a model polycyclic aromatic hydrocarbon (PAH), fluoranthene. The sediment was then validated with two other model PAHs, benz[a]anthracene and benzo[a]pyrene. Various developmental end points were recorded, including survival, embryonic heartbeat, hatching delay, hatching success, larval biometry and abnormalities. The final artificial sediment composition was set at 2.5 % dry weight (dw) Sphagnum peat, 5 % dw kaolin clay and 92.5 % dw silica of 0.2- to 0.5-mm grain size. In contrast with natural sediments, the chemical components of this artificial matrix are fully defined and readily identifiable. It is totally safe for fish embryos and presents relatively high sorption capacities for hydrophobic compounds. Studies with other hydrophobic and metallic contaminants and mixtures should be performed to further validate this artificial sedimen

Most persistent organic pollutants, due to their hydrophobic properties, accumulate in aquatic sediments and represent a high risk for sediment quality. To assess the toxicity of hydrophobic pollutants, a novel approach was recently proposed as an alternative to replace, refine and reduce animal experimentation: the medaka embryo-larval sediment contact assay (MELAc). This assay is performed with Japanese medaka embryos incubated on a natural sediment spiked with the compound being tested. With the aim of improving this assay, our study developed a reference exposure protocol with an artificial sediment specifically designed to limit natural sediment composition uncertainties and preparation variability. The optimum composition of the new artificial sediment was tested using a model polycyclic aromatic hydrocarbon (PAH), fluoranthene. The sediment was then validated with two other model PAHs, benz[a]anthracene and benzo[a]pyrene. Various developmental end points were recorded, including survival, embryonic heartbeat, hatching delay, hatching success, larval biometry and abnormalities. The final artificial sediment composition was set at 2.5 % dry weight (dw) Sphagnum peat, 5 % dw kaolin clay and 92.5 % dw silica of 0.2- to 0.5-mm grain size. In contrast with natural sediments, the chemical components of this artificial matrix are fully defined and readily identifiable. It is totally safe for fish embryos and presents relatively high sorption capacities for hydrophobic compounds. Studies with other hydrophobic and metallic contaminants and mixtures should be performed to further validate this artificial sediment.

Most persistent organic pollutants, due to their hydrophobic properties, accumulate in aquatic sediments and represent a high risk for sediment quality. To assess the toxicity of hydrophobic pollutants, a novel approach was recently proposed as an alternative to replace, refine and reduce animal experimentation: the medaka embryo–larval sediment contact assay (MELAc). This assay is performed with Japanese medaka embryos incubated on a natural sediment spiked with the compound being tested. With the aim of improving this assay, our study developed a reference exposure protocol with an artificial sediment specifically designed to limit natural sediment composition uncertainties and preparation variability. The optimum composition of the new artificial sediment was tested using a model polycyclic aromatic hydrocarbon (PAH), fluoranthene. The sediment was then validated with two other model PAHs, benz[a]anthracene and benzo[a]pyrene. Various developmental end points were recorded, including survival, embryonic heartbeat, hatching delay, hatching success, larval biometry and abnormalities. The final artificial sediment composition was set at 2.5 % dry weight (dw) Sphagnum peat, 5 % dw kaolin clay and 92.5 % dw silica of 0.2- to 0.5-mm grain size. In contrast with natural sediments, the chemical components of this artificial matrix are fully defined and readily identifiable. It is totally safe for fish embryos and presents relatively high sorption capacities for hydrophobic compounds. Studies with other hydrophobic and metallic contaminants and mixtures should be performed to further validate this artificial sediment.