Laurent Peluhet

Tidal influence on the distribution of hydrophobic organic contaminants in the Seine Estuary and biomarker responses on the copepod Eurytemora affinis

To elucidate tidally related variations of hydrophobic organic contaminant (HOC) bioavailability and the impact of these contaminants on estuarine ecosystems, both PCB and PAH concentrations were investigated in the dissolved phase and in the suspended particulate material (SPM) of the Seine Estuary. Both PAH and PCB highest levels were observed in surface and bottom water when SPM remobilizations were maximum, in relation to higher speed currents. In parallel, acetylcholinesterase (AChE) and glutathione-S-transferase (GST) activities were investigated in the copepod Eurytemora affinis. Significant decreasing AChE levels were measured during the tidal cycle and between surface and bottom copepods related to salinity and to HOC concentration variations. Significant increasing GST levels were also observed when HOC concentrations in the water column were the highest. This study underlined the need to standardize sampling procedures for biomonitoring studies in order to avoid interfering factors that could modify biomarker responses to chemical exposure.

Are exploited mangrove molluscs exposed to Persistent Organic Pollutant contamination in Senegal, West Africa?

The surface sediments, two bivalves (Arca senilis and Crassostera gasar) and three gastropods (Conus spp., Hexaplex duplex and Pugilina morio) from two Senegalese stations, Falia (Sine-Saloum Estuary) and Fadiouth (Petite Côte), were analyzed for their pollutant organic persistent contamination (polychlorinated biphenyls PCBs; organochlorinated pesticides OCPs; polybrominated diphenyl ethers PBDEs). Results revealed significant levels of PCBs, DDTs and lindane in mangrove sediments ranging from 0.3 to 19.1, 0.3 to 15.9, and 0.1 to 1.9 ng g(-1) d.w., respectively. Among the other POPs analysed, only hexachlorobenzene, heptachlor and trans-nonachlor for OCPs, as well as BDE47 and BDE99 congeners for PBDEs were detected at very low concentrations, generally not of concern. POP levels and patterns were in good accordance with literature data available for other tropical developing countries. A seasonal quantitative difference was highlighted with higher levels of PCBs and DDTs in sediments after the wet season, likely due to the strong wash-out of residues from inland to the marine ecosystems during the rainy season. The observed pattern of DDT and its metabolites pointed out probable recent applications of DDT for public health emergencies in Senegal. Exploited molluscs were exposed to the same POP compounds as those measured in sediments. They presented OCP levels within the same range as in sediments, while significant higher concentrations of PCBs were observed in shellfish soft tissues revealing a higher bioaccumulation potential mainly due to the lipophilicity of these compounds. Finally, the influence of the reproduction cycle on POP levels through lipid content variations was highlighted, minimizing potential differences in POP bioaccumulation between shellfish species. From an ecotoxicological and public health point of view, results from this study revealed that POPs in sediments from the Petite Côte and the Sine-Saloum Estuary would not cause toxic effects and impairments in molluscs from these regions, and that no potential risk exists for human, especially local populations, through mangrove shellfish consumption.

Wandering albatrosses document latitudinal variations in the transfer of persistent organic pollutants and mercury to Southern Ocean predators.

Top marine predators are effective tools to monitor bioaccumulative contaminants in remote oceanic environments. Here, we used the wide-ranging wandering albatross Diomedea exulans to investigate potential geographical variations of contaminant transfer to predators in the Southern Ocean. Blood concentrations of 19 persistent organic pollutants and 14 trace elements were measured in a large number of individuals (N = 180) of known age, sex and breeding status from the subantarctic Crozet Islands. Wandering albatrosses were exposed to a wide range of contaminants, with notably high blood mercury concentrations. Contaminant burden was markedly influenced by latitudinal foraging habitats (inferred from blood δ(13)C values), with individuals feeding in warmer subtropical waters having lower concentrations of pesticides, but higher concentrations of mercury, than those feeding in colder subantarctic waters. Sexual differences in contaminant burden seemed to be driven by gender specialization in feeding habitats, rather than physiological characteristics, with females foraging further north than males. Other individual traits, such as adult age and reproductive status, had little effect on blood contaminant concentrations. Our study provides further evidence of the critical role of global distillation on organic contaminant exposure to Southern Ocean avian predators. In addition, we document an unexpected high transfer of mercury to predators in subtropical waters, which merits further investigation.

A new spiked sediment assay using embryos of the Japanese medaka specifically designed for a reliable toxicity assessment of hydrophobic chemicals

Despite their low water solubility, hydrophobic pollutants are widespread in the aquatic environment and could represent a threat for living organisms. EU regulations on chemicals require accurate and reliable data on chemical toxicity. Current normalised fish toxicity assays, in particular those advocated by OECD guidelines, do not allow reliable toxicity assessment of hydrophobic compounds due to their low water solubility. In order to accurately evaluate the toxicity of this kind of compounds, a new spiked sediment assay using embryos of the Japanese medaka was developed. It consists of directly exposing fertilised eggs, during their entire embryonic development, onto the reference sediment spiked with the test compound. A large set of lethal or sublethal effects in embryos and newly hatched larvae, including non-invasive endpoints is analysed in order to maximise the sensitivity of the test. The approach was validated using four model pollutants with different modes of action: DMBA, PCB126, PCB153 and 4-nonylphenol (NP). All compounds, except PCB153, induced a dose-dependent increase in toxic effects. In fact, lethal effects only occurred at the highest tested concentration. In contrast, sub-lethal effects including skeletal deformations, cardiac activity modulation, body length reduction and hatching delay were observed at low to moderate concentrations of DMBA and PCB126. NP induced subtle effects in embryos, altering cardiac activity and hatching success but only at high concentrations. Although a few more improvements would make it a fully standardised assay, this spiked sediment assay using medaka embryos proves to be sensitive enough to measure hydrophobic chemical toxicity using an environmentally realistic mode of exposure.

Biliary PAH metabolites, EROD activity and DNA damage in dab (Limanda limanda) from Seine Estuary (France)

The Seine Estuary is well known to be widely contaminated by organic pollutants and especially by polycyclic aromatic hydrocarbons (PAHs). Fish are known to metabolize PAHs, leading to different toxic effects at both cellular and sub-cellular levels. In this work, we studied the relationships between the 7-ethoxyresorufin-O-deethylase (EROD) activity in the liver, the level of DNA strand breaks in blood cells and the concentration of PAH metabolites in the bile of the sentinel flatfish species Limanda limanda. Muscle and liver samples were analysed for parent PAH levels. Female and male dabs of two size classes (juveniles and adults) were collected by trawling in two sites with different degrees of pollution during March and September 2005 and 2006. Significant effects of sex, age, site and season were demonstrated on EROD activity and the level of strand breaks. Parent PAH concentrations in dabs did not allow discriminating of the two sampling sites. However, for PAH metabolites, significant differences were observed with sites and seasons. Dabs collected at the mouth of the estuary appeared to be the most impacted when looking at the results obtained with the three selected markers. The significant correlations observed between the level of PAH metabolites and the level of DNA lesions showed the importance of a combined analysis of chemical and biochemical markers to correctly assess the contribution of chemical contamination to the toxic effects measured in situ in fish.

Transcriptome profile analysis reveals specific signatures of pollutants in Atlantic eels

Identifying specific effects of contaminants in a multi-stress field context remain a challenge in ecotoxicology. In this context, “omics” technologies, by allowing the simultaneous measurement of numerous biological endpoints, could help unravel the in situ toxicity of contaminants. In this study, wild Atlantic eels were sampled in 8 sites presenting a broad contamination gradient in France and Canada. The global hepatic transcriptome of animals was determined by RNA-Seq. In parallel, the contamination level of fish to 8 metals and 25 organic pollutants was determined. Factor analysis for multiple testing was used to identify genes that are most likely to be related to a single factor. Among the variables analyzed, arsenic (As), cadmium (Cd), lindane (γ-HCH) and the hepato-somatic index (HSI) were found to be the main factors affecting eel’s transcriptome. Genes associated with As exposure were involved in the mechanisms that have been described during As vasculotoxicity in mammals. Genes correlated with Cd were involved in cell cycle and energy metabolism. For γ-HCH, genes were involved in lipolysis and cell growth. Genes associated with HSI were involved in protein, lipid and iron metabolisms. Our study proposes specific gene signatures of pollutants and their impacts in fish exposed to multi-stress conditions.

The mussel caging approach in assessing biological effects of wastewater treatment plant discharges in the Gulf of Finland (Baltic Sea)

Biological effects of wastewater treatment plant (WWTP) effluents were investigated in Baltic mussels (Mytilus trossulus) caged for one month 800m and 1100m from the WWTP discharge site and at a reference site 4km away. Significant antioxidant, genotoxic and lysosomal responses were observed close to the point of the WWTP discharge. Passive samplers (POCIS) attached to the cages indicated markedly higher water concentrations of various pharmaceuticals at the two most impacted sites. Modeling the dispersal of a hypothetical passive tracer compound from the WWTP discharge site revealed differing frequencies and timing of the exposure periods at different caging sites. The study demonstrated for the first time the effectiveness of the mussel caging approach in combination with passive samplers and the application of passive tracer modeling to examine the true exposure patterns at point source sites such as WWTP pipe discharges in the Baltic Sea.

Abnormal Ovarian DNA Methylation Programming during Gonad Maturation in Wild Contaminated Fish

There is increasing evidence that pollutants may cause diseases via epigenetic modifications. Epigenetic mechanisms such as DNA methylation participate in the regulation of gene transcription. Surprisingly, epigenetics research is still limited in ecotoxicology. In this study, we investigated whether chronic exposure to contaminants experienced by wild female fish (Anguilla anguilla) throughout their juvenile phase can affect the DNA methylation status of their oocytes during gonad maturation. Thus, fish were sampled in two locations presenting a low or a high contamination level. Then, fish were transferred to the laboratory and artificially matured. Before hormonal treatment, the DNA methylation levels of the genes encoding for the aromatase and the receptor of the follicle stimulating hormone were higher in contaminated fish than in fish from the clean site. For the hormone receptor, this hypermethylation was positively correlated with the contamination level of fish and was associated with a decrease in its transcription level. In addition, whereas gonad growth was associated with an increase in DNA methylation in fish from the clean site, no changes were observed in contaminated fish in response to hormonal treatment. Finally, a higher gonad growth was observed in fish from the reference site in comparison to contaminated fish.

Heavy metals and organochlorinated compounds in the European eel (Anguilla anguilla) from the Adour estuary and associated wetlands (France)

Heavy metals and organic pollutants were investigated in the Adour estuary (South West France) and associated wetlands using the European eel (Anguilla anguilla) as a bioindicator. Heavy metals (Cu, Cd, Zn, Pb, and Ag) were measured in soft tissue of yellow eels. Mercury (total Hg and MeHg) and organochlorinated compounds (7 PCBs, 11 OCPs) were analysed in muscle. Concentrations in muscle were in agreement with moderately contaminated environments in Europe and were below the norms fixed for eel consumption for heavy metals and OCPs. Analyses of liver showed a higher pressure of Ag and Zn in the downstream estuary than in the freshwater sites whereas Cd was lower in the estuary probably because of the salinity influence. According to quality classes 100% of eels from freshwater sites indicated clean or slightly polluted environments. However, total mercury concentrations were close to the thresholds fixed by the European Community in the downstream estuary, whereas the sum of PCBs was found to be greatly above the fixed value. 100% of the individuals from the estuary were classified in quality classes corresponding to polluted or highly polluted sites. These first results highlight the need of further investigations focused on mercury and PCBs in this area taking the seasonal temperature influence into account for a better understanding of the pollution distribution and the possible threat on the eel population from the Adour basin.

Demographic, endocrine and behavioral responses to mirex in the South polar skua

Population consequences of chronic exposure to multiple pollutants at low environmental doses remain speculative, because of the lack of appropriate long-term monitoring surveys. This study integrates proximate and ultimate aspects of persistent organic pollutants (POP) burden in free-living vertebrates, by coupling hormonal and behavioral endpoints, life-history traits, and population dynamics. Blood samples (N=70) were collected in South polar skuas during two breeding periods, in 2003 and 2005, and individuals were annually monitored until 2011. Multi-state mark recapture models were used to test the effects of POP levels on demographic traits. Survival rate and long-term breeding probability were not related to individual POP levels, whereas long-term breeding success significantly decreased with increasing blood levels of mirex, an organochlorine insecticide. At the proximate level, corticosterone (stress hormone) and prolactin (parental care hormone) levels were not linked to individual POP burden. Nest defense in 2005 was significantly less intensive in chick-rearing skuas bearing higher mirex levels, suggesting reproductive behavioral impairment. Matrix population models were then built to project the rate of population decline according to increasing mirex burden. Although mirex levels were 2.8 times higher in 2003 than in 2005, the population-level effect of mirex was only detected in 2005, the year of higher corticosterone levels. The combination of endocrine traits with demographic analysis thereby enables to provide new support of synergistic interactions between pollutants and stress levels on long-term breeding outputs and population dynamics.