[Departement_IRSTEA]Eaux [TR1_IRSTEA]QUASARE | International audience | Detecting and separating specific effects of contaminants in a multi-stress field context remain a major challenge in ecotoxicology. In this context, the aim of this study was to assess the usefulness of a non-invasive transcriptomic method, by means of a complementary DNA (cDNA) microarray comprising 1000 candidate genes, on caudal fin clips. Fin gene transcription patterns of European eels (Anguilla anguilla) exposed in the laboratory to cadmium (Cd) or a polychloro-biphenyl (PCBs) mixture but also of wild eels from three sampling sites with differing contamination levels were compared to test whether fin clips may be used to detect and discriminate the exposure to these contaminants. Also, transcriptomic profiles from the liver and caudal fin of eels experimentally exposed to Cd were compared to assess the detection sensitivity of the fin transcriptomic response. A similar number of genes were differentially transcribed in the fin and liver in response to Cd exposure, highlighting the detection sensitivity of fin clips. Moreover, distinct fin transcription profiles were observed in response to Cd or PCB exposure. Finally, the transcription profiles of eels from the most contaminated site clustered with those from laboratory-exposed fish. This study thus highlights the applicability and usefulness of performing gene transcription assays on non-invasive tissue sampling in order to detect the in situ exposure to Cd and PCBs in fish.

Detecting and separating specific effects of contaminants in a multi-stress field context remain a major challenge in ecotoxicology. In this context, the aim of this study was to assess the usefulness of a non-invasive transcriptomic method, by means of a complementary DNA (cDNA) microarray comprising 1000 candidate genes, on caudal fin clips. Fin gene transcription patterns of European eels (Anguilla anguilla) exposed in the laboratory to cadmium (Cd) or a polychloro-biphenyl (PCBs) mixture but also of wild eels from three sampling sites with differing contamination levels were compared to test whether fin clips may be used to detect and discriminate the exposure to these contaminants. Also, transcriptomic profiles from the liver and caudal fin of eels experimentally exposed to Cd were compared to assess the detection sensitivity of the fin transcriptomic response. A similar number of genes were differentially transcribed in the fin and liver in response to Cd exposure, highlighting the detection sensitivity of fin clips. Moreover, distinct fin transcription profiles were observed in response to Cd or PCB exposure. Finally, the transcription profiles of eels from the most contaminated site clustered with those from laboratory-exposed fish. This study thus highlights the applicability and usefulness of performing gene transcription assays on non-invasive tissue sampling in order to detect the in situ exposure to Cd and PCBs in fish.

International audience | Consumption of water polluted by faecal contaminants is responsible for 2 million deaths annually, most of which occur in developing countries without adequate sanitation. In tropical aquatic systems, streambeds can be reservoirs of persistent pathogenic bacteria and high rainfall can lead to contaminated soils entering streams and to the resuspension of sediment-bound microbes in the streambed. Here, we present a novel method using fallout radionuclides (Be-7 and Pb-210(xs)) to estimate the proportions of Escherichia coli, an indicator of faecal contamination, associated with recently eroded soil particles and with the resuspension of streambed sediments. We show that using these radionuclides and hydrograph separations we are able to characterize the proportion of particles originating from highly contaminated soils and that from the resuspension of particle-attached bacteria within the streambed. We also found that although overland flow represented just over one tenth of the total flood volume, it was responsible for more than two thirds of the downstream transfer of E. coli. We propose that data obtained using this method can be used to understand the dynamics of faecal indicator bacteria (FIB) in streams thereby providing information for adapted management plans that reduce the health risks to local populations.

Consumption of water polluted by faecal contaminants is responsible for 2 million deaths annually, most of which occur in developing countries without adequate sanitation. In tropical aquatic systems, streambeds can be reservoirs of persistent pathogenic bacteria and high rainfall can lead to contaminated soils entering streams and to the resuspension of sediment-bound microbes in the streambed. Here, we present a novel method using fallout radionuclides (⁷Be and ²¹⁰Pbₓₛ) to estimate the proportions of Escherichia coli, an indicator of faecal contamination, associated with recently eroded soil particles and with the resuspension of streambed sediments. We show that using these radionuclides and hydrograph separations we are able to characterize the proportion of particles originating from highly contaminated soils and that from the resuspension of particle-attached bacteria within the streambed. We also found that although overland flow represented just over one tenth of the total flood volume, it was responsible for more than two thirds of the downstream transfer of E. coli. We propose that data obtained using this method can be used to understand the dynamics of faecal indicator bacteria (FIB) in streams thereby providing information for adapted management plans that reduce the health risks to local populations. Graphical Abstract Graphical abstract showing (1) the main water flow processes (i.e. overland flow, groundwater return flow, blue arrows) and sediment flow components (i.e. resuspension and soil erosion, black arrows) during floods in the Houay Pano catchment; (2) the general principle of the method using fallout radionuclide markers (i.e. ⁷Be and ²¹⁰Pbₓₛ) to estimate E. coli load from the two main sources (i.e. streambed resuspension vs soil surface washoff); and 3) the main results obtained during the 15 May 2012 storm event (i.e. relative percentage contribution of each process to the total streamflow, values in parentheses)

International audience | Phthalates are ubiquitous contaminants and endocrine-disrupting chemicals that can become trapped in the cuticles of insects, including ants which were recognized as good bioindicators for such pollution. Because phthalates have been noted in developed countries and because they also have been found in the Arctic, a region isolated from direct anthropogenic influence, we hypothesized that they are widespread. So, we looked for their presence on the cuticle of ants gathered from isolated areas of the Amazonian rainforest and along an anthropogenic gradient of pollution (rainforest vs. road sides vs. cities in French Guiana). Phthalate pollution (mainly di(2-ethylhexyl) phthalate (DEHP)) was higher on ants gathered in cities and along road sides than on those collected in the pristine rainforest, indicating that it follows a human-mediated gradient of disturbance related to the use of plastics and many other products that contain phthalates in urban zones. Their presence varied with the ant species; the cuticle of Solenopsis saevissima traps higher amount of phthalates than that of compared species. However, the presence of phthalates in isolated areas of pristine rainforests suggests that they are associated both with atmospheric particles and in gaseous form and are transported over long distances by wind, resulting in a worldwide diffusion. These findings suggest that there is no such thing as a "pristine" zone.

Phthalates are ubiquitous contaminants and endocrine-disrupting chemicals that can become trapped in the cuticles of insects, including ants which were recognized as good bioindicators for such pollution. Because phthalates have been noted in developed countries and because they also have been found in the Arctic, a region isolated from direct anthropogenic influence, we hypothesized that they are widespread. So, we looked for their presence on the cuticle of ants gathered from isolated areas of the Amazonian rainforest and along an anthropogenic gradient of pollution (rainforest vs. road sides vs. cities in French Guiana). Phthalate pollution (mainly di(2-ethylhexyl) phthalate (DEHP)) was higher on ants gathered in cities and along road sides than on those collected in the pristine rainforest, indicating that it follows a human-mediated gradient of disturbance related to the use of plastics and many other products that contain phthalates in urban zones. Their presence varied with the ant species; the cuticle of Solenopsis saevissima traps higher amount of phthalates than that of compared species. However, the presence of phthalates in isolated areas of pristine rainforests suggests that they are associated both with atmospheric particles and in gaseous form and are transported over long distances by wind, resulting in a worldwide diffusion. These findings suggest that there is no such thing as a “pristine” zone.

International audience | Burkholderia pseudomallei is the bacterium that causes melioidosis in humans. While B. pseudomallei is known to be endemic in South East Asia (SEA), the occurrence of the disease in other parts of the tropics points towards a potentially large global distribution. We investigated the environmental factors that influence the presence (and absence) of B. pseudomallei in a tropical watershed in SEA. Our main objective was to determine whether there is a link between the presence of the organism in the hydrographic network and the upstream soil and land-use type. The presence of B. pseudomallei was determined using a specific quantitative real-time PCR assay following enrichment culture. Land use, soil, geomorphology, and environmental data were then analyzed using partial least squares discriminant analysis (PLSDA) to compare the B. pseudomallei positive and negative sites. Soil type in the surrounding catchment and turbidity had a strong positive influence on the presence (acrisols and luvisols) or absence (ferralsols) of B. pseudomallei. Given the strong apparent links between soil characteristics, water turbidity, and the presence/absence of B. pseudomallei, actions to raise public awareness about factors increasing the risk of exposure should be undertaken in order to reduce the incidence of melioidosis in regions of endemicity.

Burkholderia pseudomallei is the bacterium that causes melioidosis in humans. While B. pseudomallei is known to be endemic in South East Asia (SEA), the occurrence of the disease in other parts of the tropics points towards a potentially large global distribution. We investigated the environmental factors that influence the presence (and absence) of B. pseudomallei in a tropical watershed in SEA. Our main objective was to determine whether there is a link between the presence of the organism in the hydrographic network and the upstream soil and land-use type. The presence of B. pseudomallei was determined using a specific quantitative real-time PCR assay following enrichment culture. Land use, soil, geomorphology, and environmental data were then analyzed using partial least squares discriminant analysis (PLSDA) to compare the B. pseudomallei positive and negative sites. Soil type in the surrounding catchment and turbidity had a strong positive influence on the presence (acrisols and luvisols) or absence (ferralsols) of B. pseudomallei. Given the strong apparent links between soil characteristics, water turbidity, and the presence/absence of B. pseudomallei, actions to raise public awareness about factors increasing the risk of exposure should be undertaken in order to reduce the incidence of melioidosis in regions of endemicity.

International audience | AbstractThis study addresses the spatio-temporal dynamics of per and polyfluoroalkyl substances (PFASs) in a highly urbanized freshwater hydrosystem, the Seine River (NW France). The distribution of PFASs between water, sediment, and periphytic biofilm was investigated at three sampling sites along a longitudinal gradient upstream and downstream from the Paris urban area. Seasonal variability was assessed through four sampling campaigns performed under contrasting hydrological conditions. In the dissolved phase, ∑PFASs fluctuated between 2 and 9 ng L−1 upstream and 6–105 ng L−1 downstream from Paris. Negative correlations between dissolved PFAS levels and river flow rate were generally observed, corroborating the predominance of point-source PFAS inputs at these sites. 18/19 target PFASs were detected, with a predominance of PFHxS and PFOS (20% of ∑PFASs each), except for the farthest downstream site where 6:2 FTSA was prevalent (35 ± 8% of ∑PFASs), likely reflecting industrial and urban inputs. In biofilms, ∑PFASs fell in the 4–32 ng g−1 dw range, and substantial bioconcentration factors (BCFs) were reported for PFNA, PFDA, and PFOS (log BCF 2.1–4.3), higher than those of PFHxS or PFOA. BCFs varied inversely with dissolved PFAS levels, potentially pointing to concentration-dependent bioaccumulation. Biofilm community characteristics (C/N ratio) may also be an influential determinant of PFAS bioaccumulation.

International audience | AbstractThis study addresses the spatio-temporal dynamics of per and polyfluoroalkyl substances (PFASs) in a highly urbanized freshwater hydrosystem, the Seine River (NW France). The distribution of PFASs between water, sediment, and periphytic biofilm was investigated at three sampling sites along a longitudinal gradient upstream and downstream from the Paris urban area. Seasonal variability was assessed through four sampling campaigns performed under contrasting hydrological conditions. In the dissolved phase, ∑PFASs fluctuated between 2 and 9 ng L−1 upstream and 6–105 ng L−1 downstream from Paris. Negative correlations between dissolved PFAS levels and river flow rate were generally observed, corroborating the predominance of point-source PFAS inputs at these sites. 18/19 target PFASs were detected, with a predominance of PFHxS and PFOS (20% of ∑PFASs each), except for the farthest downstream site where 6:2 FTSA was prevalent (35 ± 8% of ∑PFASs), likely reflecting industrial and urban inputs. In biofilms, ∑PFASs fell in the 4–32 ng g−1 dw range, and substantial bioconcentration factors (BCFs) were reported for PFNA, PFDA, and PFOS (log BCF 2.1–4.3), higher than those of PFHxS or PFOA. BCFs varied inversely with dissolved PFAS levels, potentially pointing to concentration-dependent bioaccumulation. Biofilm community characteristics (C/N ratio) may also be an influential determinant of PFAS bioaccumulation.

International audience | Microbial communities driving the nitrogen cycle contribute to ecosystem services such as crop production and air, soil, and water quality. The responses to herbicide stress of ammonia-oxidizing and ammonia-denitrifying microbial communities were investigated by an analysis of changes in structure-function relationships. Their potential activities, abundances (quantitative PCR), and genetic structure (denaturing gradient gel electrophoresis) were assessed in a microcosm experiment. The application rate (1 × FR, 0.45 μg g(-1) soil) of the mesotrione herbicide did not strongly affect soil N-nutrient dynamics or microbial community structure and abundances. Doses of the commercial product Callisto® (10 × FR and 100 × FR) or pure mesotrione (100 × FR) exceeding field rates induced short-term inhibition of nitrification and a lasting stimulation of denitrification. These effects could play a part in the increase in soil ammonium content and decrease in nitrate contents observed in treated soils. These functional impacts were mainly correlated with abundance shifts of ammonia-oxidizing Bacteria (AOB) and Archaea (AOA) or denitrifying bacteria. The sustained restoration of nitrification activity, from day 42 in the 100 × FR-treated soils, was likely promoted by changes in the community size and composition of AOB, which suggests a leading role, rather than AOA, for soil nitrification restoration after herbicide stress. This ecotoxicological community approach provides a nonesuch multiparameter assessment of responses of N-cycling microbial guilds to pesticide stress.

Microbial communities driving the nitrogen cycle contribute to ecosystem services such as crop production and air, soil, and water quality. The responses to herbicide stress of ammonia-oxidizing and ammonia-denitrifying microbial communities were investigated by an analysis of changes in structure-function relationships. Their potential activities, abundances (quantitative PCR), and genetic structure (denaturing gradient gel electrophoresis) were assessed in a microcosm experiment. The application rate (1 × FR, 0.45 μg g⁻¹ soil) of the mesotrione herbicide did not strongly affect soil N-nutrient dynamics or microbial community structure and abundances. Doses of the commercial product Callisto® (10 × FR and 100 × FR) or pure mesotrione (100 × FR) exceeding field rates induced short-term inhibition of nitrification and a lasting stimulation of denitrification. These effects could play a part in the increase in soil ammonium content and decrease in nitrate contents observed in treated soils. These functional impacts were mainly correlated with abundance shifts of ammonia-oxidizing Bacteria (AOB) and Archaea (AOA) or denitrifying bacteria. The sustained restoration of nitrification activity, from day 42 in the 100 × FR-treated soils, was likely promoted by changes in the community size and composition of AOB, which suggests a leading role, rather than AOA, for soil nitrification restoration after herbicide stress. This ecotoxicological community approach provides a nonesuch multiparameter assessment of responses of N-cycling microbial guilds to pesticide stress.

Avec nos remerciements à Jodie Thénard, Virginie Grondin, Jean-Pierre Pétraud, Amélie Trouvé, Christelle Marrauld et Françoise Poiroux | Earthworms play a key role in agroecosystem soil processes. This study aims to assess the effects of different doses of a commercial formulation of epoxiconazole (Opus®), a persistent and widely used fungicide, on the earthworm Aporrectodea icterica. A laboratory study was conducted in a natural soil in order to measure effects of Opus® on earthworm mortality, uptake, weight gain, enzymatic activities (catalase and glutathione-S-transferase), and energy resources (lipids and glycogens). The estimated LC50 was 45.5 mg kg−1, or 268 times the recommended dose. Weight gains were 28, 19, and 13 % of the initial weight after 28 days of exposure in the control and D1 and D10 (1 and 10 times the recommended dose) treatments, respectively. No difference was observed for catalase activity between the three treatments, at 7, 14, or 28 days. The glutathion-S-transferase (GST) activity was two times as high in D1 as in D0 at 14 days. At 28 days, glycogen concentration was lower in D10 than in the D1 treatment. This study highlighted moderate sublethal effects of the commercial formulation Opus® for earthworms. Considering that these effects were observed on a species found in cultivated fields, even at recommended rates, much more attention should be paid to this pesticide.

Avec nos remerciements à Jodie Thénard, Virginie Grondin, Jean-Pierre Pétraud, Amélie Trouvé, Christelle Marrauld et Françoise Poiroux | Earthworms play a key role in agroecosystem soil processes. This study aims to assess the effects of different doses of a commercial formulation of epoxiconazole (Opus®), a persistent and widely used fungicide, on the earthworm Aporrectodea icterica. A laboratory study was conducted in a natural soil in order to measure effects of Opus® on earthworm mortality, uptake, weight gain, enzymatic activities (catalase and glutathione-S-transferase), and energy resources (lipids and glycogens). The estimated LC50 was 45.5 mg kg−1, or 268 times the recommended dose. Weight gains were 28, 19, and 13 % of the initial weight after 28 days of exposure in the control and D1 and D10 (1 and 10 times the recommended dose) treatments, respectively. No difference was observed for catalase activity between the three treatments, at 7, 14, or 28 days. The glutathion-S-transferase (GST) activity was two times as high in D1 as in D0 at 14 days. At 28 days, glycogen concentration was lower in D10 than in the D1 treatment. This study highlighted moderate sublethal effects of the commercial formulation Opus® for earthworms. Considering that these effects were observed on a species found in cultivated fields, even at recommended rates, much more attention should be paid to this pesticide.