International audience | Multiple reaction monitoring (MRM) mass spectrometry is emerging as a relevant tool for measuring customized molecular markers in freshwater sentinel species. While this technique is typically used for the validation of protein molecular markers preselected from shotgun experiments, recent gains of MRM multiplexing capacity offer new possibilities to conduct large-scale screening of animal proteomes. By combining the strength of active biomonitoring strategies and MRM technologies, this study aims to propose a new strategy for the discovery of candidate proteins that respond to environmental variability. For this purpose, 249 peptides derived from 147 proteins were monitored by MRM in 273 male gammarids caged in 56 environmental sites, representative of the diversity of French water bodies. A methodology is here proposed to identify a set of customized housekeeping peptides (HKPs) used to correct analytical batch effects and allow proper comparison of peptide levels in gammarids. A comparative analysis performed on HKPs-normalized data resulted in the identification of peptides highly modulated in the environment and derived from proteins likely involved in the environmental stress response. Overall, this study proposes a breakthrough approach to screen and identify potential proteins responding to relevant environmental conditions in sentinel species.

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International audience | Numerous toxicological studies have shown that ingestion of pesticides can induce physiological stress in breeding birds, with adverse consequences on egg laying parameters and offspring quality through parental effects. However, previous studies do not mimic current levels of pesticide residues in typical landscapes, and they do not consider potential cocktail effects of pesticides as they occur in the wild. Herein, we explored whether realistic pesticide exposure affected reproduction parameters and offspring condition through parental effects in Grey partridge. We fed 24 breeding pairs with either seeds from conventional agriculture crops treated with various pesticides during cropping, or organic grains without pesticide residues as controls. The conventional and organic grain diets mimicked food options potentially encountered by wild birds in the field. The results showed that ingesting low pesticide doses over a long period had consequences on reproduction and offspring quality without altering mortality in parents or chicks. Compared with organic pairs, conventional pairs yielded smaller chicks at hatching that had a lower body mass index at 24 days old. Additionally, these chicks displayed lower haematocrit when body mass index was higher. Therefore, ingestion of conventional grains by parents resulted in chronic exposure to pesticide residues, even at low doses, and this had detrimental consequences on offspring. These results demonstrate a sublethal effect of pesticide residues through parental effects. The consequences of parental exposure on chicks might partly explain the decline in wild Grey partridge populations, which raises questions for avian conservation and demography if current agrosystem approaches are continued

International audience | The impact of persistent organic pollutants (POPs) on reproductive health is still poorly understood, even though infertility management has high associated societal and economical costs. The aims of this study were to characterize the internal levels of polychlorinated biphenyls (PCBs), organochlorine pesticides (OCPs) and perfluoroalkylated substances (PFAS) in women undergoing in vitro fertilization (IVF); and evaluate their association with IVF outcomes, individually and as mixtures in a combined multipollutant approach. Thus, 136 women undergoing IVF treatment at Nantes University Hospital (France) were prospectively recruited between 2019 and 2020. Serum samples were analyzed using liquid chromatography with tandem-mass spectrometry for 14 PFAS. Follicular fluid was analyzed with gas chromatography coupled to high resolution mass spectrometry for 14 PCBs and 25 OCPs. Intermediate and clinical IVF outcomes were ascertained by embryologists and clinicians using standardized protocols. Multivariate Poisson regression models and Bayesian Kernel Machine Regressions (BKMR) were used to identify individual and joint associations between POPs and IVF outcomes adjusting for age, body mass index (BMI) and anti-Müllerian hormone. The results showed that most POPs were widely present in women, and globally not associated with clinically relevant IVF outcomes, like live birth rates. Nonetheless, negative associations between PCB138 and trans-nonachlor with useable blastocysts were identified, β -0.28 (95%CI [-0.52; -0.04] p = 0.02) and β -0.22 (95%CI [-0.40; -0.03] p = 0.02). Conversely, PCB28 showed positive associations with the number of useable blastocysts, pregnancy rate and live birth rate. The BKMR analysis suggested the lack of association of the mixture with intermediate and clinical outcomes. The study supports the need of conducting further studies in a larger population sample in order to ensure sufficient statistical power to identify modest effects and a robust stratification analysis to account for the large underlying disease heterogeneity.

International audience | Carbamazepine (CBZ) and Hg are widespread and persistent micropollutants in aquatic environments. Both pollutants are known to trigger similar toxicity mechanisms, e.g. reactive oxygen species (ROS) production. Here, their effects were assessed in the zebra mussel Dreissena polymorpha, frequently used as a freshwater model in ecotoxicology and biomonitoring. Single and co-exposures to CBZ (3.9 µg•L-1) and MeHg (280 ng•L-1) were performed for 1 and 7 days. Metabolomics analyses evidenced that the co-exposure was the most disturbing after 7 days, reducing the amount of 25 metabolites involved in protein synthesis, energy metabolism, antioxidant response and osmoregulation, and significantly altering cells and organelles' structure supporting a reduction of functions of gills and digestive glands. CBZ alone after 7 days decreased the amount of α-aminobutyric acid and had a moderate effect on the structure of mitochondria in digestive glands. MeHg alone had no effect on mussels' metabolome, but caused a significant alteration of cells and organelles' structure in gills and digestive glands. Single exposures and the co-exposure increased antioxidant responses vs control in gills and digestive glands, without resulting in lipid peroxidation, suggesting an increased ROS production caused by both pollutants. Data globally supported that a higher number of hyperactive cells compensated cellular alterations in the digestive gland of mussels exposed to CBZ or MeHg alone, while CBZ+MeHg co-exposure overwhelmed this compensation after 7 days. Those effects were unpredictable based on cellular responses to CBZ and MeHg alone, highlighting the need to consider molecular toxicity pathways for a better anticipation of effects of pollutants in biota in complex environmental conditions.

International audience | Chlordecone (CLD), was widely applied in banana fields in the French West Indies from 1972 to 1993. The WISORCH model was constructed to assess soil contamination by CLD and estimated that it lasts from 100 to 600 years, depending on leaching intensity and assuming no degradation. However, recent studies demonstrated that CLD is degraded in the environment, hence questioning the reliability of previous estimations. This paper shows how to improve the model and provides insights into the long-term dissipation of CLD. In-situ observations were made in nearly 2545 plots between 2001 and 2020, and 17 plots were sampled at two dates. Results of soil analyses showed an unexpected 4-fold decrease in CLD concentrations in the soil, in contrast to simulations made using the first version of WISORCH at the time. Neither erosion, nor CLD leaching explained these discrepancies. In a top-down modeling approach, these new observations of CLD concentrations led us to implement a new dissipation process in the WISORCH model that corresponds to a DT50 dissipation half-life of 5 years. The new version of the improved model allowed us to update the prediction of the persistence of soil pollution, with soil decontamination estimated for the 2070s. This development calls for re-evaluation of soil pollution status. Further validation of the new version of WISORCH is needed so it can contribute to crop management on contaminated soil.

International audience | The intestinal epithelium is a functional and physical barrier formed by a cell monolayer that constantly differentiates from a stem cell in the crypt. This is the first target for food contaminants, especially mycotoxins. Deoxynivalenol (DON) is one of the most prevalent mycotoxins. This study compared the effects of DON (0-100 μM) on proliferative and differentiated intestinal epithelial cells. Three cell viability assays (LDH release, ATP content and neutral red uptake) indicated that proliferative Caco-2 cells are more sensitive to DON than differentiated ones. The establishment of transepithelial electrical resistance (TEER), as a read out of the differentiation process, was delayed in proliferative cells after exposure to 1 μM DON. Transcriptome analysis of proliferative and differentiated exposure to 0-3 μM DON for 24 h revealed 4862 differentially expressed genes (DEG) and indicated an effect of both the differentiation status and the DON treatment. KEGG enrichment analysis indicated involvement of metabolism, ECM receptors and tight junctions in the differentiation process, while ribosome biogenesis, mRNA surveillance, and the MAPK pathway were involved in the response to DON. The number of differentially expressed genes and the amplitude of the effect were higher in proliferative cells exposed to DON than that in differentiated cells. In conclusion, our study shows that proliferative cells are more susceptible than differentiated ones to DON and that the mycotoxin delays the differentiation process.

International audience | In utero exposure to environmental chemicals, such as synthetic phenols, may alter DNA methylation in different tissues, including placenta - a critical organ for fetal development. We studied associations between prenatal urinary biomarker concentrations of synthetic phenols and placental DNA methylation. Our study involved 202 mother-son pairs from the French EDEN cohort. Nine phenols were measured in spot urine samples collected between 22 and 29 gestational weeks. We performed DNA methylation analysis of the fetal side of placental tissues using the IlluminaHM450 BeadChips. We evaluated methylation changes of individual CpGs in an adjusted epigenome-wide association study (EWAS) and identified differentially methylated regions (DMRs). We performed mediation analysis to test whether placental tissue heterogeneity mediated the association between urinary phenol concentrations and DNA methylation. We identified 46 significant DMRs (>= 5 CpGs) associated with triclosan (37 DMRs), 2,4-dichlorophenol (3), benzophenone-3 (3), methyl- (2) and propylparaben (1). All but 2 DMRs were positively associated with phenol concentrations. Out of the 46 identified DMRs, 7 (6 for triclosan) encompassed imprinted genes (APC, FOXG1, GNAS, GNASAS, MIR886, PEG10, SGCE), which represented a significant enrichment. Other identified DMRs encompassed genes encoding proteins responsible for cell signaling, transmembrane transport, cell adhesion, inflammatory, apoptotic and immunological response, genes encoding transcription factors, histones, tumor suppressors, genes involved in tumorigenesis and several cancer risk biomarkers. Mediation analysis suggested that placental cell heterogeneity may partly explain these associations. This is the first study describing the genome-wide modifications of placental DNA methylation associated with pregnancy exposure to synthetic phenols or their precursors. Our results suggest that cell heterogeneity might mediate the effects of triclosan exposure on placental DNA methylation. Additionally, the enrichment of imprinted genes within the DMRs suggests mechanisms by which certain exposures, mainly to triclosan, could affect fetal development.

International audience | It has been proposed that non-protein thiols and organic acids play a major role in cadmium phytoavailability and distribution in plants. In the Cd-hyperaccumulator Solanum nigrum and non-accumulator Solanum melongena, the role of these organic ligands in the accumulation and detoxification mechanisms of Cd are debated. In this study, we used X-ray absorption spectroscopy to investigate Cd speciation in these plants (roots, stem, leaves) and in the soils used for their culture to unravel the plants responses to Cd exposure. The results show that Cd in the 100 mg.kg-1 Cd-doped clayey loam soil is sorbed onto iron oxyhydroxides. In both S. nigrum and S. melongena, Cd in roots and fresh leaves is mainly bound to thiol ligands, with a small contribution of inorganic S ligands in S. nigrum leaves. We interpret the Cd binding to sulfur ligands as detoxification mechanisms, possibly involving the sequestration of Cd complexed with glutathione or phytochelatins in the plant vacuoles. In the stems, results show an increase binding of Cd to -O ligands (>50% for S. nigrum). We suggest that Cd is partly complexed by organic acids for transportation in the sap.

International audience | Climate warming is expected to impact the response of species to insecticides. Recent studies show that this interaction between insecticides and temperature can depend on other factors. Here, we tested for the influence of transgenerational effects on the Insecticide x Temperature interaction in the crop pest moth Spodoptera littoralis. Specifically, we analysed reaction norms among experimental clutches based on a split-plot design crossing the factors temperature, insecticide and clutch. The study was performed on 2280 larvae reared at four temperatures (23, 25, 27 and 29°C), and their response to the insecticide deltamethrin (three concentrations and a control group) was tested. Temperature had a global influence with effects on larval survival, duration of development, pupal body mass, and significant reaction norms of the clutches for temperature variations of only 2°C. In addition to the expected effect of deltamethrin on mortality, the insecticide slightly delayed the development of S. littoralis, and the effects on mortality and development differed among the clutches. Projection models integrating all the observed responses illustrated the additive effects of deltamethrin and temperature on the population multiplication rate. Variation in the response of the clutches showed that transgenerational effects influenced the impact of insecticide and temperature. Although no evidence indicated that the Insecticide x Temperature interaction depended on transgenerational effects, the studies on the dependence of the Insecticide x Temperature interaction on other factors continue to be crucial to confidently predict the combined effects of insecticides and climate warming.