[Departement_IRSTEA]Eaux [TR1_IRSTEA]BELCA<br/>EA ECOLDUR CT3 | International audience | Microorganisms are ubiquitous in soil, air, and water ecosystems, where they are key players of ecosystem services. Microbial ecotoxicology is an emerging interdisciplinary area of research which aims at investigating the impact of human activities on the diversity, abundance, and activity of microorganisms. In return, the results of such investigations hold the promise to provide novel ways of assessing in a sensitive way the impacts of diverse environmental disturbances and subsequent ecosystem responses. Thus and although the term itself is yet rarely encountered in the scientific literature, microbial ecotoxicology already addresses an increasing political and societal demand. In the French scientific landscape, which often mimics the famous (but sometimes indigestible) “mille-feuilles” pastry, microbial ecotoxicologists are scattered across many different research centers belonging to different research organizations and universities. This research field has thus lacked any visibility and remained unorganized until now. Formal organization of scientific activities may be considered a typical “froggies” concern (or ailment). Nevertheless, it is rather surprising that scientific journals and significant international conferences specifically devoted to microbial ecotoxicology have been missing so far, especially considering the plethoric range of journals and congresses devoted to microbial ecology and ecotoxicology. With these considerations in mind, the idea of organizing the French research community of microbial ecologists around concepts of ecotoxicology made its way, with the aim of sharing the necessity to overcome artificial boundaries that prevent progress in this promising field.

Polycyclic aromatic hydrocarbons (PAHs) are a diverse family of more than one hundred compounds, containing at least two aromatic rings. In addition to parent compounds, the PAH family also includes substituted derivatives, bearing one or several alkyl groups, sulfur, or oxygen. In the environment, PAHs are ubiquitous and present as very complex mixtures. They can also be associated with metallic and/or other organic compounds. The composition of PAH mixtures depends on their origin. There are two major types of such PAH mixtures, petrogenic and pyrogenic, which enter the environment through different routes. Petrogenic mixtures originate from oils, including natural oil seeps. They enter the aquatic environment due to harbor activity or soil runoff or as a consequence of oil spills. Pyrolytic mixtures result from the incomplete combustion of organic matter, including fossil fuel, entering aquatic environments through deposits of atmospheric emissions directly into water or soil, followed by soil erosion and runoff. Directly linked to human activity, the release of PAHs into the environment has increased over the last few decades. As an example, the amount of PAHs released into the atmosphere has dramatically increased from under 50,000 tons in 1987 (Eisler 1987) to over 500,000 tons in 2004 (Zhang and Tao 2009)

[Departement_IRSTEA]Ecotechnologies [TR1_IRSTEA]TED | International audience | This study set out to assess how vertical flow constructed wetlands (vfCW) adapt to different types of tourism-driven variations in influent load, i.e. (i) campsites, closed for six months of the year and featuring a two-month-only window of high-season activity, and (ii) tourist-interest villages either hosting tourists over at least the six months of summer while the permanent population is resident or summer festivals that create intense pollution loads in a short burst of just a few days. The study surveyed 4 campsites and 4 tourist-interest villages that were monitored for several years, generating over 70 performance balances for vfCW that were intentionally scaled down in relation to the conventional French design for experimental trials. The influent wastewater effectively qualifies as domestic sewage, although relatively concentrated, with the campsites presenting particularly high nitrogen concentrations (122 gTKN.L-1). The applied daily loads were also particularly high, with some combinations of load parameters (hydraulic load, organic matter, TKN) leading to 400% overloading. Even under these drastic conditions, quality of effluent remained excellent on the characteristic organic matter parameters, with removal performances always over 85%. However, nitrification performances were poor to good. Analysis of the dataset points to two major design thresholds: For campsites, in order to keep a 73% nitrification rate even at the height of the summer season, the load applied to the 1st stage filter in operation has to be capped at less than 600 gCOD.m-².d-1. For tourist-interest villages, in order to keep an 85% nitrification rate in the summer season, the load applied to the 2nd stage filter in operation has to be capped at less than 22 gTKN.m-².d-1. Here, vfCW were demonstrated to robustly handle a massive increase in loads applied, providing the wetland construction and operation stringently follow best design standards and practices.

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.

eISSN: 1614-7499 | International audience | Buffer zones such as ponds and ditches are used to reduce field-scale losses of pesticides from subsurface drainage waters to surface waters. The objective of this study was to assess the efficiency of these buffer zones, in particular constructed wetlands, focusing specifically on sorption processes. We modelled the sorption processes of three herbicides [2-methyl-4-chlorophenoxyacetic acid (2,4-MCPA), isoproturon and napropamide] and three fungicides (boscalid, prochloraz and tebuconazole) on four substrates (two soils, sediment and straw) commonly found in a pond and ditch in Lorraine (France). A wide range of Freundlich coefficient (K (fads)) values was obtained, from 0.74 to 442.63 mg(1 -aEuro parts per thousand n) L (n) kg(-1), and the corresponding K (foc) values ranged from 56 to 3,725 mg(1 -aEuro parts per thousand n) L (n) kg(-1). Based on potential retention, the substrates may be classified as straw >> sediments > soils. These results show the importance of organic carbon content and nature in the process of sorption. Similarly, the studied pesticides could be classified according to their adsorption capacity as follows: prochloraz >> tebuconazole-boscalid > napropamide >> MCPA-isoproturon. This classification is strongly influenced by the physico-chemical properties of pesticides, especially solubility and K (oc). Straw exhibited the largest quantity of non-desorbable pesticide residues, from 12.1 to 224.2 mg/L for all pesticides. The presence of plants could increase soil-sediment sorption capacity. Thus, establishment and maintenance of plants and straw filters should be promoted to optimise sorption processes and the efficiency of ponds and ditches in reducing surface water pollution.

In aquatic environments, polycyclic aromatic hydrocarbons (PAHs) mostly occur as complex mixtures, for which risk assessment remains problematic. To better understand the effects of PAH mixture toxicity on fish early life stages, this study compared the developmental toxicity of three PAH complex mixtures. These mixtures were extracted from a PAH-contaminated sediment (Seine estuary, France) and two oils (Arabian Light and Erika). For each fraction, artificial sediment was spiked at three different environmental concentrations roughly equivalent to 0.5, 4, and 10 μg total PAH g−1 dw. Japanese medaka embryos were incubated on these PAH-spiked sediments throughout their development, right up until hatching. Several endpoints were recorded at different developmental stages, including acute endpoints, morphological abnormalities, larvae locomotion, and genotoxicity (comet and micronucleus assays). The three PAH fractions delayed hatching, induced developmental abnormalities, disrupted larvae swimming activity, and damaged DNA at environmental concentrations. Differences in toxicity levels, likely related to differences in PAH proportions, were highlighted between fractions. The Arabian Light and Erika petrogenic fractions, containing a high proportion of alkylated PAHs and low molecular weight PAHs, were more toxic to Japanese medaka early life stages than the pyrolytic fraction. This was not supported by the toxic equivalency approach, which appeared unsuitable for assessing the toxicity of the three PAH fractions to fish early life stages. This study highlights the potential risks posed by environmental mixtures of alkylated and low molecular weight PAHs to early stages of fish development.

Polycyclic aromatic hydrocarbons (PAHs) are ubiquitous contaminants that can be present at high levels as mixtures in polluted aquatic environments. Many PAHs are potent mutagens and several are well-known carcinogens. Despite numerous studies on individual compounds, little is known about the toxicity of PAHs mixtures that are encountered in environmental situations. In the presentwork, zebrafish were continuously fed from 5 days post-fertilisation to 14 months post-fertilisation (mpf) with a diet spiked with fractions of either pyrolytic (PY), petrogenic light oil (LO), or petrogenic heavy oil (HO) origin at three concentrations. A decrease in survival was identified after 3 mpf in fish fed with the highest concentration of HO or LO, but not for PY. All PAH fractions caused preneoplastic and neoplastic disorders in longterm-exposed animals. Target tissues were almost exclusively of epithelial origin, with the bile duct epithelium being themost susceptible to chronic exposure to all PAH fractions, and with germ cells being the second most responsive cells. Significantly higher incidences of neoplasms were observed with increasing PAH concentration and exposure duration. Themost severe carcinogenic effects were induced by dietary exposure to HO compared to exposure to LO or PY (45, 30 and 7 %, respectively, after 9 to 10 months of exposure to an intermediate concentration of PAHs). In contrast, earliest carcinogenic effects were detected as soon as 3 mpf after exposure to LO, including the lowest concentration, or to PY. PAHbioactivation and genotoxicity in blood was assessed by ethoxyresorufin-Odeethylase activity quantification and comet and micronuclei assays, respectively, but none of these were positive. Chronic dietary exposure of zebrafish to PAH mixtures results in carcinogenotoxic events that impair survival and physiology of exposed fish.

A natural sediment spiked with three individual polycyclic aromatic hydrocarbons (PAHs; pyrene, phenanthrene and benzo[a]pyrene) was used to expose zebrafish embryos and larvae during 4 days. The total PAH concentration was 4.4 μg g−1 which is in the range found in sediment from contaminated areas. Quantification of metabolites in the larvae after exposure confirmed the actual contamination of the larvae and indicated an active metabolism specially for pyrene and benzo[a]pyrene. After a transfer in a clean medium, the larvae were reared to adulthood and valuated for survival, growth, reproduction, and behavior. Measured endpoints revealed a late disruption of growth (appearing at 5 months) and a trend toward a lower reproductive ability. Adults of embryos exposed to sediment spiked with PAHs displayed lethargic and/or anxiety-like behaviors. This latter behavior was also identified in offspring at larval stage. All together, these effects could have detrimental consequences on fish performances and contribution to 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.

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.