Despite their intensive use and their impact on ecosystems, biocides and surfactants are still poorly regulated and poorly monitored at large scale. In the frame of the revision of the national regulatory surveillance plan of surface waters, France planned in 2018 a monitoring campaign at national scale focused on these two types of substances of very emerging concern. Forty-nine contaminants (32 biocides and 17 surfactants) were investigated in surface water and sediment samples from 91 sampling sites, and in effluent and sludge samples of 7 wastewater treatment plants (WWTP), in mainland France and overseas regions. Between 33 and 52 % of the target contaminants were quantified at least once in water and sediment. High frequencies of quantification were observed for the surfactants (up to 91 % in water samples and up to 57 % in sediment samples for LAS C10-C13) and for the biocides (up to 64 % for fipronil in water samples and up to 90 % for methyl nonyl ketone in sediment samples). The median concentrations of surfactants were up to 2 μg/L in mainland surface water samples and up to 528 μg/kg in sediment samples, and for biocides, the median concentrations were up to 0.18 μg/L in mainland surface water samples and up to 104 μg/kg in sediment samples. PNEC exceedances in water and sediment were determined for both types of substances. The analysis of effluent and sludge suggested significant but not total removal of these substances in the WWTP. Temporal and spatial variations of the concentrations of both types of substances in surface water samples were also observed, suggesting both punctual and diffuse contamination sources of the surface water investigated.

L’analyse non ciblée connaît une forte période de croissance dans le domaine de l’analyse environnementale, car elle ouvre le champ des investigations de la qualité des milieux à un très grand nombre de composés. Cet article consiste en un retour d’expérience de 12 laboratoires français sur les pratiques et les précautions à considérer lors de la réalisation d’analyses non ciblées, en particulier pour la recherche de composés suspects dans les échantillons issus des milieux aquatiques. En effet, si cette technique apporte des informations nouvelles et très prometteuses, elle n’en reste pas moins une technique complexe qu’il convient d’utiliser avec de nombreuses précautions pour assurer des résultats de qualité. La réflexion commune entreprise dans le groupe de laboratoires, a permis de synthétiser des informations techniques afin de présenter les enjeux de cette méthodologie d’intérêt majeur en analyse environnementale de façon simple et accessible aux laboratoires, demandeurs d’analyse ou toute personne intéressée par ce type d’approche. Les verrous liés à la spécificité des analyses non ciblées, au développement de méthodes et au rendu de résultats ont été développés pour fournir des clés de compréhension pour les laboratoires et les utilisateurs de résultats. Ce travail s’attache ainsi à être un guide de découverte de la technique pour les personnes désireuses de voir ce que peut apporter l’analyse non ciblée, mais également un guide de bonnes pratiques pour celles et ceux qui acquièrent ou qui analysent des données issues de ces techniques.

For decades, oil extraction in rural sites in the North Amazon Region (NAR) in Ecuador, have generated mixtures of potentially toxic compounds, such as polycyclic aromatic hydrocarbons (PAHs) and metal(loid)s. The main national refinery and the thermal power plant located in Esmeraldas, on the North Pacific Coast (NPC), are also considered as important sources of air contamination. Particulate matter (PM10) emitted at both sites could induce the formation of reactive oxygen species (ROS) in the lungs upon inhalation and could be associated with respiratory diseases. In this study, PM10 mass composition was monitored over a two-year period in both regions: NAR (close to oil platforms and open flares) and NPC (in a public school close to the refinery). PM10 composition was assessed in terms of metal(loid)s, organic and elementary carbon (OC, EC), monosaccharides (levoglucosan, mannosan, galactosan), glucose, polyols (sorbitol, mannitol, arabitol), water soluble ions and polycyclic aromatic compounds (PAHs, oxy-PAHs and nitro-PAHs). Additionally, three complementary biochemical and acellular tests were performed to evaluate the oxidative potential (OP). Results show that the PM10 mass and elemental concentrations were higher in NPC than in NAR. Barium and Mo concentrations, commonly used in oil operations, were up to 1000-fold higher than values recorded in other regions of Ecuador. OC/EC ratios and polyols concentrations were higher in NAR than in NPC, indicating a larger biogenic contribution to the PM mass in this region. In NAR, the main sources associated with ROS burden were biogenic emissions and oil production, as indicated by positive correlations between OP, sugars, Ba, some PAHs and oxy-PAHs. On the other hand, in NPC, associations between NH4+, Ba, As and Ni imply that oil refining and industrial activities are the main contributors to the OP of PM10.

La preservation de la qualite de l'eau est un enjeu majeur a la fois pour l'environnement et pour la sante humaine. Dans le contexte de la Directive Cadre sur l'Eau (DCE, 2000/60/CE), de nombreux travaux ont etudie les micropolluants organiques, et les connaissances concernant leur presence, voies d'introduction, sources et impacts sur les ecosystemes aquatiques ont considerablement augmente. Les eaux usees etant des sources importantes de micropolluants en lien avec la consommation de produits manufactures, l'application de la DCE a conduit au renforcement de la reglementation sur le traitement des eaux usees urbaines et a la generalisation des procedes biologiques tels que les boues activees a aeration prolongee ou les biofiltres, permettant d'eliminer significativement les micropolluants organiques. Les processus d'oxydation impliques consistent en une degradation qui peut ne pas etre complete, generant des produits de transformation (TP) relativement stables et toxiques susceptibles de se retrouver a la fois dans les boues et effluents liquides. Jusqu'a present, les donnees sur les TP restent rares et se limitent a quelques composes. Ainsi, il apparait essentiel que les futurs projets de recherche portent sur la question des TP afin d'elucider leur presence, formation et devenir tout au long du systeme de traitement des eaux usees jusqu'a leur entree potentielle dans le milieu aquatique par le biais des rejets d'effluents des stations d'epuration (STEP). Objet ou objectifs TRANSPRO etudie la formation des TP, en developpant des methodes innovantes de screening utilisant des outils a la fois chimiques (Spectrometrie de Masse Haute Resolution) et biologiques (tests in vitro). Il etudie l'ensemble du systeme de traitement des eaux usees (des entrees de STEP) jusqu'aux ecosystemes aquatiques naturels, en se concentrant sur differents types de procedes de traitement par rapport a leur capacite a generer des TP ainsi que sur les procedes naturels (biodegradation, photo-oxydation) pouvant donner lieu a des transformations dans le milieu lui-meme. Methode TRANSPRO est un projet finance par l'agence nationale de la recherche (ANR) sur la periode 2019-2022. Il est porte par un consortium de partenaires publics. C'est un projet collaboratif impliquant un partenariat solide base sur une expertise pluridisciplinaire, associant chimistes analyticiens (EPOC), physico-chimistes (EPOC, LGC), chimistes de l'environnement (EPOC, Irstea), specialistes en genie des procedes et modelisation (Irstea, LGC), (eco) toxicologues (EPOC), specialistes des systemes de traitement des eaux usees (LGC, Irstea), tous rassembles pour aborder une question commune: quels processus generent des TP, que sont ces TP et quels sont ceux pertinents d'un point de vue environnemental ? Resultats TRANSPRO va permettre d'ameliorer nos connaissances sur la nature, l'origine et la dynamique des TP. Il permettra egalement de classifier les procedes de traitement des eaux usees par rapport a leur tendance a generer des TP et d'aider a selectionner le processus le plus efficace en termes de degradation des contaminants parents mais minimisant la formation des TP. TRANSPRO fournira ainsi des connaissances qui contribueront a ameliorer les traitements des eaux usees pour concevoir les systemes d'assainissement de demain. Tout savoir sur le projet TRANSPRO : https://lnkd.in/dkZnUyS

The present study analysed potential adverse effects of discontinuous sources of contamination, namely the discharge of a combined sewer overflow (CSO) and of runoff in an urban area, the Bay of Santander (North Iberian Peninsula). Water samples and caged mussels were used to analyse concentrations of contaminants and biological responses. Mussels (Mytilus galloprovincialis) were transplanted to a marina receiving runoff from a petrol station and to a CSO discharge site. Samples were collected in synchrony with heavy rains along 62days. Lysosomal membrane stability (LMS) and acyl-CoA oxidase (AOX) activity were measured as core biomarkers and were analysed at all sampling times. Histopathology of digestive gland and gonads, transcription levels of vitellogenin gene, volume density of black silver deposits and micronuclei formation were measured at initial and final stages of the transplant. Chemical analyses of metals, polycyclic aromatic hydrocarbons (PAHs) and endocrine disruptors were performed in water samples and mussel flesh. Mussels accumulated low concentrations of contaminants, which is in accordance with results obtained from exposure biomarkers. AOX activity decreased in all transplanted mussels after the first heavy rain, but this change seems to be related to the seasonal pattern of the enzyme activity. Mussels located close to the CSO discharge site showed a reduction in LMS after the first rain event, when compared to mussels before the transplant and to mussels from the reference location. However, this was attributable to natural environmental changes rather than to pollution. Values of the rest of analysed biomarkers were below threshold values reported for the study area.

Blue mussels (Mytilus trossulus) were transplanted in cages for three months in two Swedish coastal areas in the Bothnian Sea (northern Baltic Sea) to investigate the interactions between analysed environmental chemicals and biological responses. A wide array of biological parameters (biomarkers) including antioxidant and biotransformation activity, geno-, cyto- and neurotoxic effects, phagocytosis, bioenergetic status and heart rate were measured to detect the possible effects of contaminants. Integrated Biomarker Response index and Principal Component Analysis performed on the individual biological response data were able to discriminate between the two study areas as well as the contaminated sites from their respective local reference sites. The two contaminated sites outside the cities of Sundsvall (station S1) and Gävle (station G1) were characterised by different biomarker response patterns. Mussels at station S1 showed a low condition index, increased heart rate recovery time and phagocytosis activity coinciding with the highest tissue concentrations of some trace metals, polycyclic aromatic hydrocarbons and organotins. At station G1 the highest organochlorine pesticide concentration was recorded as well as elevations in glutathione S-transferase activity, thiamine content and low lysosomal membrane stability. Significant variability in the geno- and cytotoxic responses and bioenergetic status was also observed at the different caging stations. The results obtained suggest that different chemical mixtures present in the study areas cause variable biological response patterns in organisms.

4-Nonylphenol (4-NP) is a synthetic organic chemical able to mimick the action of the nat- ural female hormone 17 β -estradiol (E2) by binding to the E2 receptor [...]

The European legislation, and in particular the Water Framework Directive requires the development of cost efficient monitoring tools that can provide the required information for the assessment of water contamination. Passive sampling methods represent one of the novel tools that have a potential to be used in various regulatory monitoring programmes aimed at assessing the levels of chemical pollutants. These methods are particularly interesting for sampling polar organic pollutants in water because they provide representative information of the water quality over extended time periods (days to weeks) in environments with fluctuating contaminant concentrations. This is achieved by integrative sampling of pollutants over the whole sampler deployment period. These tools can be coupled to toxicity testing using bioassays that give information on toxic and ecotoxic hazards associated to substances that are present, these substances being identified or not. In this study the polar organic chemical integrative sampler (POCIS) was used in surface water to evaluate the water contamination by polar organic compounds and their potential toxicity.

An analytical procedure for the simultaneous determination of twelve endogenous steroids (testosterone, androstenedione, 17beta-estradiol, estrone, pregnenolone, progesterone, dihydroandrostenedione, dihydrotestosterone, 11alpha-ketotestosterone, 17alpha-hydroxyprogesterone, 17alpha-hydroxypregnenolone, 17alpha,20beta-dihydroxy-4-pregnen-3-one) in plasma and bile samples by solid-phase extraction (SPE) and gas chromatography/mass spectrometry (GC-MS) has been developed. After enzymatic hydrolysis for bile samples only, samples were concentrated and purified using two successive SPE (C(18) and NH(2)) cartridges. Analytes were derivatized with a mixture of N-methyl-N-(trimethylsilyl)trifluoroacetamide (MSTFA) / mercaptoethanol / ammonium iodide (NH(4)I) and determined by GC-MS in selective ion monitoring mode. For most of the steroids monitored, recoveries were in the range 90-120% in plasma and in the range 60-70% in bile, and the reproducibility was below 10% for the complete procedure. Limits of detection obtained ranged from 0.1 to 0.4 ng/g in fish plasma and from 1.6 to 14 ng/g in fish bile. The developed method was successfully applied to the determination of plasma steroids in flounders (Platichthys flesus) collected from two French estuaries.

The aim of the present work was to investigate genotoxicant accumulation and biological responses of zebra mussels and blue mussels collected along a pollution gradient in the Seine estuary and in the Seine Bay. The sampling area included three contaminated and one reference sites for each species. The study focused on polyaromatic hydrocarbons (PAH), lindane, polychlorobiphenyls (PCB) and metals known to be potential genotoxicants and/or reactive oxygen species (ROS) inducers. Enzymatic activities related to cellular defence systems including the phase II enzyme glutathione S-transferase (GST) and three antioxidant enzymes superoxide dismutase (SOD), catalase (CAT) and glutathion peroxydase (GPx) were measured in gills. DNA adducts and DNA strand breaks (Comet assay) were measured in digestive gland and hemocytes, respectively. Species differences were observed in metal accumulation (As and Pb), GPx activity and DNA adduct formation. A marked upstream–downstream gradient was reported for PAH body burden and to a lesser extent for PCB and metals with the highest values measured just downstream the industrialized area of Rouen. GST and SOD activities in gills of bivalves were positively related to PAH and metals body burden, respectively. Activation of those cellular defences may prevent accumulation of electrophilic metabolites and free radicals and thus may protect DNA and others macromolecules against oxidation and adduction. Although DNA strand breaks and bulky adducts were detected in both species, levels were relatively low and no significant site differences were observed in June 2003. Our results indicate a clear relationship between genotoxicant accumulation and positive activation of detoxification and antioxidant systems but poor consequences in term of DNA damage for wild population of mussels inhabiting the Seine estuary.

The purpose of this study was to examine PAH accumulation and bulky DNA adduct formation in the digestive gland of zebra mussels exposed in their habitat or in controlled laboratory conditions to complex mixture of PAH. DNA adducts were measured using a 32P-postlabelling protocol with nuclease P1 enrichment adapted from Reddy and Randerath [Reddy, M.V., Randerath, K., 1986. Nuclease P1-mediated enhancement of sensitivity of 32P-postlabelling test for structurally diverse DNA adducts. Carcinogenesis 7, 1543–1551]. Specimens collected in the upper part of the Seine estuary were shown to accumulate higher levels of PAH (up to 1.6μgg−1 dry weight) in comparison to individuals from the reference site (0.053μgg−1 dry weight). The former exhibited elevated levels of DNA adducts (up to 4.0/108 nucleotides) and higher diversity of individual adducts with five distinct spots being specifically detected in individuals originating from the Seine estuary. Zebra mussels exposed for 5 days to 0.01% (v/v) of organic extract of sediment from the Seine estuary were shown to accumulate high amounts of PAH (up to 138μgg−1 dry weight) but exhibited relatively low levels of DNA adducts. Exposure to benzo[a]pyrene led to a dose-dependent accumulation of B[a]P (up to 7063μgg−1 dry weight) and a clear induction of DNA adduct formation in the digestive gland of mussels (up to 1.13/108 nucleotides). Comparisons with other bivalves exposed to the same model PAH, revealed similar levels of adducts and comparable adduct profiles with a main adduct spot and a second faint one. This study clearly demonstrated that zebra mussels are able to biotransform B[a]P and probably other PAH into reactive metabolites with DNA-binding activity. This work also demonstrated the applicability of the nuclease P1 enhanced 32P-postlabelling method for bulky adduct detection in the digestive gland of zebra mussels. DNA adduct measurement in zebra mussels could be a suitable biomarker to monitor PAH-exposure and evaluate genotoxicity in fresh water ecosystems.

In order to characterize the genotoxicity in the Seine estuary and Seine bay, chemical and toxicological analyses were performed on 17 sediments collected in June 2001 and June 2003. Many potent mutagenic and/or carcinogenic compounds – including polycyclic aromatic hydrocarbons (PAH), polychlorinated biphenyls (PCB) and metals – were detected. Those compounds were found to be at relatively high concentrations in the upper part of the Seine estuary but were barely detectable at sites outside the plume from the Seine. The levels of pollution did not vary significantly between the two sampling periods, except that PAH concentrations in sediments collected at Oissel and Le Havre showed a marked increase in June 2003. The toxicity of organic extracts from sediments was evaluated by both embryotoxicity and in vitro genotoxicity (SOS Chromotest) assays. Organic extracts from sediments taken from the Seine estuary appeared significantly more embryotoxic than those from the Seine bay. In addition, the sediment extracts from the upper part of the Seine estuary exhibited higher genotoxicity than those from the lower part, and no genotoxicity was reported for sediments from the Seine bay. The genotoxic activity was detected only after adding an S9 microsomal fraction, suggesting the preponderant involvement of pro-genotoxic organic compounds. In addition, SOS Chromotest responses obtained with purified organic fractions revealed that PAH and, to a lesser extent, unknown polar organic compounds were probably responsible for this genotoxicity. Altogether, these results suggest that sediments from the upper Seine estuary are genotoxic and embryotoxic, and therefore, could be potentially hazardous for species living or feeding in the area.

Marine mussels Mytilus sp. were transplanted on a monthly basis in cages over one year to oyster farms and harbours in the Arcachon Bay (France) in order to assess the water quality of the bay. Contaminant levels (organotin compounds, trace metals, PCBs and PAHs) were measured in tissues of transplanted mussels and mussels from a reference station, along with physiological parameters of the mussels (condition indexes, lipid content and dry weight). Four biomarkers (AChE: acetylcholinesterase activity, GST: gluthathione S-transferase activity, CAT: catalase activity and TBARS: thiobarbituric acid-reactive substance content) were also monitored. The remote stations monitored (oyster parks) exhibited no accumulation pattern of pollutants. Their respective concentrations therefore constitute a background level of the contamination in the bay ([TBT]= 30 ng Sn g(-1) dw, [SigmaHAPs]= 100 ng g(-1) dw, [SigmaPCBs]= 35 ng g(-1) dw). The elevated chemical contamination of the largest harbour of the bay, the Arcachon harbour, can be interpreted in terms of persistence of organotin compounds ([SigmaOTs]= 1500-2000 ng Sn g(-1) dw) and PAHs ([SigmaHAPs]= 4500-5000 ng g(-1) dw) in sediments and, to a lesser extent, of direct inputs of copper ([Cu]= 20 microg g(-1) dw in harbours versus 7 in oyster parks) and petrogenic PAHs ([methylphenanthrenes]= 1600 ng g(-1) dw in the dockyard versus 170 at the gas stations), related to the use of copper-based antifouling paints and to dockyard activity, respectively. However, the Arcachon Bay presents a low contamination level by PCBs and metals, including harbour stations. Furthermore, higher levels of other PAHs (particularly alkyl PAHs such as methylphenanthrenes/1600 ng g(-1) dw) not included in the 16 PAHs from the EPA priority list (usually studied in biomonitoring programmes/1500 ng g(-1) dw) in the Arcachon harbour underline the need to integrate these compounds in biomonitoring of highly PAH-polluted areas such as harbours in order to avoid misinterpretation of the biological responses observed. Biomarker responses were not able to discriminate the different chemical contamination levels recorded in the Arcachon Bay and rather reflected changes in environmental factors. Furthermore, the strong intraspecies variability of biological responses could be due to genetic differences of mussels from the Arcachon Bay. It is the first time that such an integrated monitoring is performed in the Arcachon Bay, also taking into account seasonal variations of chemical contents and biomarkers levels in mussel tissues.

Blue mussels, Mytilus edulis, were exposed to two different concentrations of tributyltin (TBT) in seawater, 1,000 ng Sn/L (C1 experiment) and 10 ng Sn/L (C2 experiment), for 4 d, in order to evaluate the bioaccumulation of TBT by mussels Mytilus edulis in microcosms and to test the ability of a multimarkers analysis to determine the effects of TBT on the biochemical parameters in mussels. Tissue burdens of Mytilus edulis were 204 +/- 7 and 2,120 +/- 4 ng Sn/g TBT after the 4-d tests for the C2 and C1 experiments, respectively. Analyses of dissected organs and/or tissues demonstrated that TBT accumulated to the greatest extent in gills in the C1 experiment and in the digestive gland in the C2 experiment. Bioconcentration factors (BCFs) were 12,100 +/- 300 and 2,000 +/- 10 for mussels exposed in the C2 and C1 experiments, respectively. The four biomarkers used in this work were acetylcholinesterase (AChE), glutathione S-transferase (GST), catalase (CAT) activities, and thiobarbituric acid-reactive substances (TBARS) contents. No significant changes were observed in the measured enzyme activities or in TBARS concentration after the 4-d TBT exposure.