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Health indicators and contaminant levels of a critically endangered species in the Gironde estuary, the European sturgeon
2020
Acolas, Marie-Laure | Davail, Blandine | Gonzalez, Patrice | Jean, Séverine | Clérandeau, Christelle | Morin, Bénédicte | Gourves, Pierre-Yves | Daffe, Guillemine | Labadie, Pierre | Perrault, Annie | Lauzent, Mathilde | Pierre, Maud | Le Barh, Romaric | Baudrimont, Magalie | Peluhet, Laurent | Le Menach, Karyn | Budzinski, Hélène | Rochard, Eric | Cachot, Jérôme | Ecosystèmes aquatiques et changements globaux (UR EABX) ; Institut National de Recherche pour l’Agriculture, l’Alimentation et l’Environnement (INRAE) | Environnements et Paléoenvironnements OCéaniques (EPOC) ; Observatoire aquitain des sciences de l'univers (OASU) ; Université Sciences et Technologies - Bordeaux 1 (UB)-Institut national des sciences de l'Univers (INSU - CNRS)-Centre National de la Recherche Scientifique (CNRS)-Université Sciences et Technologies - Bordeaux 1 (UB)-Institut national des sciences de l'Univers (INSU - CNRS)-Centre National de la Recherche Scientifique (CNRS)-École Pratique des Hautes Études (EPHE) ; Université Paris sciences et lettres (PSL)-Université Paris sciences et lettres (PSL)-Centre National de la Recherche Scientifique (CNRS) | Laboratoire d'Ecophysiologie et Ecotoxicologie des Systèmes Aquatiques (LEESA) ; Université Sciences et Technologies - Bordeaux 1 (UB)-Institut Français du Pétrole-Centre National de la Recherche Scientifique (CNRS) | Laboratoire Ecologie Fonctionnelle et Environnement (LEFE) ; Institut Ecologie et Environnement - CNRS Ecologie et Environnement (INEE-CNRS) ; Centre National de la Recherche Scientifique (CNRS)-Centre National de la Recherche Scientifique (CNRS)-Université Toulouse III - Paul Sabatier (UT3) ; Université de Toulouse (UT)-Université de Toulouse (UT)-Observatoire Midi-Pyrénées (OMP) ; Institut de Recherche pour le Développement (IRD)-Université Toulouse III - Paul Sabatier (UT3) ; Université de Toulouse (UT)-Université de Toulouse (UT)-Institut national des sciences de l'Univers (INSU - CNRS)-Centre National d'Études Spatiales [Toulouse] (CNES)-Centre National de la Recherche Scientifique (CNRS)-Météo-France-Institut de Recherche pour le Développement (IRD)-Institut national des sciences de l'Univers (INSU - CNRS)-Centre National d'Études Spatiales [Toulouse] (CNES)-Centre National de la Recherche Scientifique (CNRS)-Météo-France-Centre National de la Recherche Scientifique (CNRS)-Institut National Polytechnique (Toulouse) (Toulouse INP) ; Université de Toulouse (UT) | Region Nouvelle-Aquitaine | IRSTEA | Adour-Garonne water agency | Region Nouvelle-Aquitaine | Gironde general council in 2014 | ANR-13-CESA-0018,SturTOP,Vulnérabilité et adaptabilité de la dernière population d'esturgeon européen (Acipenser sturio) aux facteurs de stress environnementaux : température, oxygène et polluants.(2013)
[Departement_IRSTEA]Eaux [TR1_IRSTEA]QUASARE [ADD1_IRSTEA]Dynamique et fonctionnement des écosystèmes | International audience | The European sturgeon, Acipenser sturio, is a highly endangered species that almost disappeared in the last decades. Thanks to yearly restocking of the population, this species is still found in the Gironde estuary (France), where juveniles grow during several years before leaving to the ocean. The aims of this study were to evaluate the pressure exerted on these fish by known organic and inorganic contaminants during their stay at the Gironde estuary, and to get information on the fish's health in this context. Monthly captures over the year 2014 provided 87 fish from the cohorts 2012 and 2013 mainly, and from cohorts 2008, 2009, and 2011, all fish born in hatchery. We report the very first analyses of contaminant levels and of biological markers measured in the blood of these fish.
Show more [+] Less [-]Health indicators and contaminant levels of a critically endangered species in the Gironde estuary, the European sturgeon
2020
Acolas, Marie-Laure | Davail, Blandine | Gonzalez, Patrice | Jean, Séverine | Clérandeau, Christelle | Morin, Bénédicte | Gourves, Pierre-Yves | Daffe, Guillemine | Labadie, Pierre | Perrault, Annie | Lauzent, Mathilde | Pierre, Maud | Le Barh, Romaric | Baudrimont, Magalie | Peluhet, Laurent | Le Menach, Karyn | Budzinski, Hélène | Rochard, Eric | Cachot, Jérôme | Ecosystèmes aquatiques et changements globaux (UR EABX) ; Institut National de Recherche pour l’Agriculture, l’Alimentation et l’Environnement (INRAE) | Environnements et Paléoenvironnements OCéaniques (EPOC) ; Observatoire aquitain des sciences de l'univers (OASU) ; Université Sciences et Technologies - Bordeaux 1 (UB)-Institut national des sciences de l'Univers (INSU - CNRS)-Centre National de la Recherche Scientifique (CNRS)-Université Sciences et Technologies - Bordeaux 1 (UB)-Institut national des sciences de l'Univers (INSU - CNRS)-Centre National de la Recherche Scientifique (CNRS)-École Pratique des Hautes Études (EPHE) ; Université Paris Sciences et Lettres (PSL)-Université Paris Sciences et Lettres (PSL)-Centre National de la Recherche Scientifique (CNRS) | Laboratoire d'Ecophysiologie et Ecotoxicologie des Systèmes Aquatiques (LEESA) ; Université Sciences et Technologies - Bordeaux 1 (UB)-Institut Français du Pétrole-Centre National de la Recherche Scientifique (CNRS) | Laboratoire Ecologie Fonctionnelle et Environnement (LEFE) ; Institut Ecologie et Environnement - CNRS Ecologie et Environnement (INEE-CNRS) ; Centre National de la Recherche Scientifique (CNRS)-Centre National de la Recherche Scientifique (CNRS)-Université Toulouse III - Paul Sabatier (UT3) ; Université de Toulouse (UT)-Université de Toulouse (UT)-Observatoire Midi-Pyrénées (OMP) ; Institut de Recherche pour le Développement (IRD)-Université Toulouse III - Paul Sabatier (UT3) ; Université de Toulouse (UT)-Université de Toulouse (UT)-Institut national des sciences de l'Univers (INSU - CNRS)-Centre National d'Études Spatiales [Toulouse] (CNES)-Centre National de la Recherche Scientifique (CNRS)-Météo-France-Institut de Recherche pour le Développement (IRD)-Institut national des sciences de l'Univers (INSU - CNRS)-Centre National d'Études Spatiales [Toulouse] (CNES)-Centre National de la Recherche Scientifique (CNRS)-Météo-France-Centre National de la Recherche Scientifique (CNRS)-Institut National Polytechnique (Toulouse) (Toulouse INP) ; Université de Toulouse (UT) | Region Nouvelle-Aquitaine | IRSTEA | Adour-Garonne water agency | Region Nouvelle-Aquitaine | Gironde general council in 2014 | ANR-13-CESA-0018,SturTOP,Vulnérabilité et adaptabilité de la dernière population d'esturgeon européen (Acipenser sturio) aux facteurs de stress environnementaux : température, oxygène et polluants.(2013)
[Departement_IRSTEA]Eaux [TR1_IRSTEA]QUASARE [ADD1_IRSTEA]Dynamique et fonctionnement des écosystèmes | International audience | The European sturgeon, Acipenser sturio, is a highly endangered species that almost disappeared in the last decades. Thanks to yearly restocking of the population, this species is still found in the Gironde estuary (France), where juveniles grow during several years before leaving to the ocean. The aims of this study were to evaluate the pressure exerted on these fish by known organic and inorganic contaminants during their stay at the Gironde estuary, and to get information on the fish's health in this context. Monthly captures over the year 2014 provided 87 fish from the cohorts 2012 and 2013 mainly, and from cohorts 2008, 2009, and 2011, all fish born in hatchery. We report the very first analyses of contaminant levels and of biological markers measured in the blood of these fish.
Show more [+] Less [-]Development in the European flounder (Platichthys flesus) of a q-PCR assay for the measurement of telomere length, a potential biomarker of pollutant effects for biomonitoring studies
2021
Akcha, Farida | Cahuc, C. | Rouxel, Julien | Munschy, Catherine | Aminot, Yann | Chouvelon, Tiphaine | Mahe, Kelig | Budzinski, H. | Mauffret, Aourell
Telomeres protect the coding sequence of chromosome ends and Telomere Length (TL) has been proposed as a biomarker of cellular aging, cumulative stress exposure and life-span in humans. With the aim to propose new biomarkers, a q-PCR protocol was adapted for the measurement of TL in the European flounder Platichthys flesus. The protocol was then applied in 2-year-old flounders from the Seine Estuary. The absolute TL in the flounder is 54 ± 13 kbp per genome (mean ± standard error). Considering relative or absolute TL, no correlation was observed with DNA damage and any of the measured contaminant concentrations (trace elements, metabolites of polycyclic aromatic hydrocarbons, polychlorobiphenyls, organochlorinated pesticides, polybrominated diphenyl ethers, perfluoroalkyl substances). Because sampling was limited, further investigations are required to state a possible impact of chemical pollution on flatfish telomeres. This is motivated by correlations observed with organochlorinated compounds when decreasing statistical significance (p ≤ 0.10).
Show more [+] Less [-]Decrease in the genotoxicity of metal-contaminated soils with biochar amendments
2017
Rees, Frédéric | Dhyèvre, Adrien | Morel, Jean-Louis | Cotelle, Sylvie | Laboratoire Sols et Environnement (LSE) ; Institut National de la Recherche Agronomique (INRA)-Université de Lorraine (UL) | Laboratoire Interdisciplinaire des Environnements Continentaux (LIEC) ; Institut Ecologie et Environnement (INEE) ; Centre National de la Recherche Scientifique (CNRS)-Centre National de la Recherche Scientifique (CNRS)-Institut national des sciences de l'Univers (INSU - CNRS)-Observatoire Terre et Environnement de Lorraine (OTELo) ; Institut national des sciences de l'Univers (INSU - CNRS)-Université de Lorraine (UL)-Centre National de la Recherche Scientifique (CNRS)-Institut national des sciences de l'Univers (INSU - CNRS)-Université de Lorraine (UL)-Centre National de la Recherche Scientifique (CNRS) | French Lorraine Region ; European Regional Development Fund
International audience | Biochar amendments, i.e., the solid product of biomass pyrolysis, reduce soil metal availability, which may lower the toxicity of metal-contaminated soils.A direct link between the decrease in soil metal availability and improved plant development is however often difficult to establish, as biochar may induce undesirable side effects on plant growth, e.g., a modification to plant nutrition. In order to investigate toxicity processes at a cellular level, roots of Vicia faba were exposed for 7 days to three metal-contaminated substrates and one control soil, amended with a 0 or 5% (w/w) addition of a wood-derived biochar.Exposure to pure biochar was also tested. Root tip cells were then observed to count the number of micronuclei as an estimation of DNA damage and the number of cells at mitosis stage. Results showed that biochar amendments led to a significant decrease in soil metal availability (Cd, Cu, Ni, Pb, and Zn) and to enhance root development on acidic substrates. The micronucleus frequency in root tip cells was positively correlated and the number of mitotic cells negatively, to the extractability of Zn in soils and to the concentration of Zn in secondary roots.Exposure to pure biochar caused a lower production of roots than most soil substrates, but led to the lowest number of observed micronuclei. In conclusion, biochar amendments can reduce the genotoxicity associated with the presence of metallic contaminants in soils, thereby potentially improving plant growth.
Show more [+] Less [-]Chronic dietary exposure of zebrafish to PAH mixtures results in carcinogenic but not genotoxic effects
2014
Larcher, Thibaut | Perrichon, P. | Vignet, Caroline | Ledevin, Mireille | Le Menach, K. | Lyphout, L. | Landi, L. | Clerandeau, C. | Le Bihanic, F. | Menard, D. | Burgeot, T. | Budzinski, H. | Akcha, F. | Cachot, J. | Cousin, Xavier | Physiopathologie Animale et bioThérapie du muscle et du système nerveux (PAnTher) ; Institut National de la Recherche Agronomique (INRA)-École nationale vétérinaire, agroalimentaire et de l'alimentation Nantes-Atlantique (ONIRIS) | PRES Université Nantes Angers Le Mans (UNAM) | École nationale vétérinaire, agroalimentaire et de l'alimentation Nantes-Atlantique (ONIRIS) | LIttoral ENvironnement et Sociétés (LIENSs) ; La Rochelle Université (ULR)-Centre National de la Recherche Scientifique (CNRS) | Laboratoire d'Écotoxicologie (LEX) ; Biogéochimie et Ecotoxicologie (BE) ; Institut Français de Recherche pour l'Exploitation de la Mer (IFREMER)-Institut Français de Recherche pour l'Exploitation de la Mer (IFREMER) | Environnements et Paléoenvironnements OCéaniques (EPOC) ; Observatoire aquitain des sciences de l'univers (OASU) ; Université Sciences et Technologies - Bordeaux 1 (UB)-Institut national des sciences de l'Univers (INSU - CNRS)-Centre National de la Recherche Scientifique (CNRS)-Université Sciences et Technologies - Bordeaux 1 (UB)-Institut national des sciences de l'Univers (INSU - CNRS)-Centre National de la Recherche Scientifique (CNRS)-École Pratique des Hautes Études (EPHE) ; Université Paris sciences et lettres (PSL)-Université Paris sciences et lettres (PSL)-Centre National de la Recherche Scientifique (CNRS) | Laboratoire de Physiologie et Génomique des Poissons (LPGP) ; Institut National de la Recherche Agronomique (INRA)-Structure Fédérative de Recherche en Biologie et Santé de Rennes ( Biosit : Biologie - Santé - Innovation Technologique ) | ANR ConPhyPop (2009-002) ; Région Poitou-Charentes ; Ifremer ; Conseil Général de Charente-Maritime ; Ministère de l'Enseignement Supérieur de la Recherche
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.
Show more [+] Less [-]Developmental toxicity of PAH mixtures in fish early life stages. Part I: adverse effects in rainbow trout
2014
Le Bihanic, Florane | Morin, Bénédicte | Cousin, Xavier | Le Menach, Karyn | Budzinski, Hélène | Cachot, Jérôme | Université Sciences et Technologies - Bordeaux 1 (UB) | Environnements et Paléoenvironnements OCéaniques (EPOC) ; Observatoire aquitain des sciences de l'univers (OASU) ; Université Sciences et Technologies - Bordeaux 1 (UB)-Institut national des sciences de l'Univers (INSU - CNRS)-Centre National de la Recherche Scientifique (CNRS)-Université Sciences et Technologies - Bordeaux 1 (UB)-Institut national des sciences de l'Univers (INSU - CNRS)-Centre National de la Recherche Scientifique (CNRS)-École Pratique des Hautes Études (EPHE) ; Université Paris sciences et lettres (PSL)-Université Paris sciences et lettres (PSL)-Centre National de la Recherche Scientifique (CNRS) | Laboratoire de Physiologie et Génomique des Poissons (LPGP) ; Institut National de la Recherche Agronomique (INRA)-Structure Fédérative de Recherche en Biologie et Santé de Rennes ( Biosit : Biologie - Santé - Innovation Technologique ) | Laboratoire d'Écotoxicologie (LEX) ; Biogéochimie et Ecotoxicologie (BE) ; Institut Français de Recherche pour l'Exploitation de la Mer (IFREMER)-Institut Français de Recherche pour l'Exploitation de la Mer (IFREMER) | ANR ConPhyPop (2009-002) ; CPER A2E ; European Regional Development Fund ; GDR EXECO ; Aquitaine Region ; Ministère de l'Enseignement Supérieur et de la Recherche
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.
Show more [+] Less [-]Developmental toxicity of PAH mixtures in fish early life stages. Part I: adverse effects in rainbow trout
2014
Le Bihanic, Florane | Morin, Benedicte | Cousin, Xavier | Le Menach, Karyn | Budzinski, Helene | Cachot, Jerome
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.
Show more [+] Less [-]Chronic dietary exposure of zebrafish to PAH mixtures results in carcinogenic but not genotoxic effects
2014
Larcher, Thibaut | Perrichon, Prescilla | Vignet, Caroline | Ledevin, Mireille | Le Menach, Karyn | Lyphout, Laura | Landi, Laure | Clerandeau, Christelle | Lebihanic, F. | Menard, Dominique | Burgeot, Thierry | Budzinski, Helene | Akcha, Farida | Cachot, J. | Cousin, Xavier
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 present work, 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 long-term-exposed animals. Target tissues were almost exclusively of epithelial origin, with the bile duct epithelium being the most 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. The most 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. PAH bioactivation and genotoxicity in blood was assessed by ethoxyresorufin-O-deethylase 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.
Show more [+] Less [-]Genotoxic and epigenetic effects of diuron in the Pacific oyster: in vitro evidence of interaction between DNA damage and DNA methylation
2021
Akcha, Farida | Barranger, Audrey | Bachère, Evelyne
Recently, research has contributed to better knowledge on the occurrence of pesticides in coastal water by identifying frequently detected substances, their concentration range and their acute and chronic toxicity for organisms. Pesticide pollution is of particular concern in France due to important agricultural activities and presence of several exoreic catchment areas that vehicle pesticides up to coastal waters, impacting non-target marine species. Several ecotoxicology questions remain to be addressed concerning the long-term effects of chronic pesticide exposure and the mechanisms involved in adaptation to chemical stress. In the present study, we brought new insights on the genetic and epigenetic effects of the herbicide diuron in oyster genitors. During gametogenesis, we exposed Crassostrea gigas to environmentally realistic herbicide concentrations (0.2–0.3 μg L−1 during two 7-day periods at half-course and end of gametogenesis). Diuron exposure was shown to decrease global DNA methylation and total methyltransferase activity in whole oyster tissue; this is consistent with the previous observation of a significant decrease in DNMT1 gene expression. Diuron effect seemed to be tissue-specific; hypermethylation was detected in the digestive gland, whereas diuron exposure had no effect on gill and gonad tissue. The genotoxicity of diuron was confirmed by the detection of one adduct in gonad DNA. By using in vitro approaches and human DNMT1 (DNMT1 has not been purified yet in bivalves), the presence of DNA lesions (adduct, 8-oxodGuo) was shown to interfere with DNMT1 activity, indicating a complex interaction between DNA damage and DNA methylation. Based on our results, we propose mechanisms to explain the effect of diuron exposure on DNA methylation, a widespread epigenetic mark.
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