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Alfonso, S., Blanc, M., Joassard, L., Keiter, S. H., Munschy, C., Loizeau, V., et al. (2019). Examining multi- and transgenerational behavioral and molecular alterations resulting from parental exposure to an environmental PCB and PBDE mixture. Aquat. Toxicol., 208, 29–38.
Résumé: Polychlorinated biphenyls (PCBs) and polybrominated diphenyl ethers (PBDEs) are persistent organic pollutants extensively used during the 20th century and still present in aquatic environments despite their ban. Effects of exposure to these compounds over generations are poorly documented. Therefore, our aims were to characterize behavioral responses and underlying molecular mechanisms in zebrafish exposed to an environmentally relevant mixture of PCBs and PBDEs as well as in four unexposed offspring generations. Zebrafish (F0) were chronically exposed from the first meal onward to a diet spiked with a mixture containing 22 PCB and 7 PBDE congeners in proportions and concentrations reflecting environmental situations (Sigma PCBs = 1991 and Sigma PBDEs = 411 ng/g). Four offspring generations (F1 to F4) were obtained from this F0 and were not further exposed. Behavior was assessed at both larval and adult stages. Mechanisms related to behavioral defects (habenula maturation and c-fos transcription) and methylation (dnmts transcription) were monitored in larvae. Exposed adult F0 as well as F1 and F3 adults displayed no behavioral change while F2 expressed anxiety-like behavior. Larval behavior was also disrupted, Le. hyperactive after light to dark transition in F1 or hypoactive in F2, F3 and F4. Behavioral disruptions may be related to defect in habenula maturation (observed in F1) and change in c-fos transcription (observed in F1 and F2). Transcription of the gene encoding DNA methyltransferase (dnmt3ba) was also modified in all generations. Our results lead us to hypothesize that chronic dietary exposure to an environmentally relevant mixture of PCB and PBDE triggers multigenerational and transgenerational molecular and behavioral disruptions in a vertebrate model.
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Benhaim, D., Ferrari, S., Colchen, T., Chatain, B., & Begout, M. - L. (2017). Relationship between individual and group learning in a marine teleost: A case study with sea bass under self-feeding conditions. Learn Behav., 45(3), 276–286.
Résumé: Fish learning and cognition are usually approached by testing single individuals in various devices such as mazes that have serious drawbacks, especially in gregarious species, including the stress induced by the test procedure. This might impair the results and lead to misinterpretation about the learning abilities of the targeted species. In order to provide an alternative to the individual-based tests, we investigated for the first time the operant conditioning of four similar groups (50 individuals per tank) of sea bass. We used two computerized self-feeder devices per tank, each coupled with individual electronic identification and that were alternately activated during varying positive appetitive reinforcement period of time (7 to 1 day). Learning abilities were examined at both group and individual levels. At the group level, the operant conditioning was demonstrated as the triggering activity significantly decreased when the device was turned off and increased when it was turned on, whatever the reinforcement period duration. The individual level analysis revealed a more complex situation with fish showing different learning performances that can be best explained through the producer-scrounger game theory.
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Cousin, X., Batel, A., Bringer, A., Hess, S., Begout, M. - L., & Braunbeck, T. (2020). Microplastics and sorbed contaminants – Trophic exposure in fish sensitive early life stages. Mar. Environ. Res., 161, 105126.
Résumé: The present study evaluated very small microplastic particle (MPs) transfer to zebrafish and marine medaka larvae via prey experimentally exposed to MPs from the onset of feeding. Larvae were fed Paramecium or Anemia nauplii loaded with fluorescent 1-5 or 10-20 mu m MP. Pollutant accumulation was analyzed by optically tracking of benzo[a]pyrene (BaP) and recording cyp1a transcription. Paramecium transferred 1-5 mu m particles only, whereas Artemia efficiently transferred both MPs. Although zebrafish and medaka larvae fed from the onset of active food intake (2-3 dph, respectively) on Paramecium and from days 6-7 post-hatch on Artemia nauplii, neither MP accumulation nor translocation to tissues was detected. MP egestion started within few hours after ingestion. Cyp1a induction and fluorescent analyses proved BaP bioavailability after transfer via Paramecium and Artemia. Unicellular or plankton organisms ingest contaminants via MPS and transfer effectively these to sensitive early life-stages of vertebrates, giving rise to whole-life exposure.
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de Oliveira, J., Chadili, E., Turies, C., Brion, F., Cousin, X., & Hinfray, N. (2021). A comparison of behavioral and reproductive parameters between wild-type, transgenic and mutant zebrafish: Could they all be considered the same “zebrafish” for reglementary assays on endocrine disruption? Comp. Biochem. Physiol. C-Toxicol. Pharmacol., 239, 108879.
Résumé: Transgenic zebrafish models are efficiently used to study the effects of endocrine disrupting chemicals (EDC); thereby informing on their mechanisms of action. However, given the reported differences between zebrafish strains at the genetical, physiological and behavioral levels; care should be taken before using these transgenic models for EDC testing. In the present study, we undertook a set of experiments in different transgenic and/or mutant zebrafish strains of interest for EDC testing: casper, cyp19a1a-eGFP, cyp19a1a-eGFP-casper, cyp11c1-eGFP, cypncl-eGFP-casper. Some behavioral traits, and some biochemical and reproductive physiological endpoints commonly used in EDC testing were assessed and compared to those obtained in WT AB zebrafish to ensure that transgene insertion and/or mutations do not negatively modify basal reproductive physiology or behavior of the fish. Behavioral traits considered as anxiety and sociality have been monitored. Sociality was evaluated by monitoring the time spent near congeners in a shuttle box while anxiety was evaluated using the Novel tank diving test. No critical difference was observed between strains for either sociality or anxiety level. Concerning reproduction, no significant difference in the number of eggs laid per female, in the viability of eggs or in the female circulating VTG concentrations was noted between the 5 transgenic/mutants and the WT AB zebrafish studied. In summary, the transgene insertion and the mutations had no influence on the endpoints measured in basal conditions. These results were a prerequisite to the use of these transgenic/mutant models for EDC testing. Next step will be to determine the sensitivity of these biological models to chemical exposure to accurately validate their use in existing fish assays for EDC testing.
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Le Bihanic, F., Clerandeau, C., Cormier, B., Crebassa, J. - C., Keiter, S. H., Beiras, R., et al. (2020). Organic contaminants sorbed to microplastics affect marine medaka fish early life stages development. Mar. Pollut. Bull., 154, 111059.
Résumé: The role of polyethylene microplastics 4-6 mu m size (MPs) in the toxicity of environmental compounds to fish early life stages (ELS) was investigated. Marine medaka Oryzias melastigma embryos and larvae were exposed to suspended MPs spiked with three model contaminants: benzo(a)pyrene (MP-BaP), perfluorooctanesulfonic acid (MP-PFOS) and benzophenone-3 (MP-BP3) for 12 days. There was no evidence of MPs ingestion but MPs agglomerated on the surface of the chorion. Fish ELS exposed to virgin MPs did not show toxic effects. Exposure to MP-PFOS decreased embryonic survival and prevented hatching. Larvae exposed to MP-BaP or MP-BP3 exhibited reduced growth, increased developmental anomalies and abnormal behavior. Compared to equivalent water-borne concentrations, BaP and PFOS appeared to be more embryotoxic when spiked on MPs than when alone in seawater. These results suggest a relevant pollutant transfer by direct contact of MPs to fish ELS that should be included in the ecotoxicological risk assessment of MPs.
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