Outils alternatifs à l'expérimentation animale pour l'évaluation de la toxicité des contaminants de l'environnement : lignées cellulaires et tests embryo-larvaires chez un poisson modèle le Médaka japonais, Oryzias latipes

Abstract : Fish are among the most widely used aquatic model for assessing the chemicalthreat before they are placed on the European market (REACh (Registration,Evaluation, Authorization and Restriction of CHemicals)). Currently, this regulation aswell as the European directive on the protection of animals used for scientific purposes(2010/63/EU) promote the establishment of alternative methods in animal testing.These methods are intended to replace animal models whenever possible, to reducethe number of animals used in experiments and to optimize the methodology appliedto animals. This thesis aims to compare the sensitivity of two of these alternative tools,cell lines and fish embryo-larval tests, using model toxicants. Then, these tools wereused to evaluate the toxicity of environmental mixtures of pollutants: microplastics andhydrocarbons (Arabian Light oil). The model compounds (B(a)P, Cd, MMS, PCB126)valid three biomarkers: reactive oxygen species (ROS) production on fish cell lines,micronucleus assay by flow cytometry and infectious challenge on Medaka larvae andjuveniles. These biomarkers combined with chemical, biochemical and physiologicalanalyzes have assessed toxicity of environmental mixtures of hydrocarbons (aqueousextract of Arabian oil = WAF) and microplastics. WAF are toxic to fish cell lines (ERODactivity induction and genotoxicity) as well as to Medaka embryos and larvae (behavior,biometrics, EROD, genotoxicity). Fish like cells seem to be particularly susceptible toArabian Light WAF. Contamination with environmental microplastics (MP) hasrevealed a toxicity of MP-associated contaminants on cell lines and fish larvae. Theingestion of plastic particles by the larvae induced lethal and sublethal effects(biometrics, behavior and EROD). The ingestion of the plastic particles by the larvaewas confirmed by two-photon microscopy. This study assessed the sensitivity andcomplementarity of these in vitro tools for assessing hazards and risks associated withchemicals.
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Pauline Pannetier. Outils alternatifs à l'expérimentation animale pour l'évaluation de la toxicité des contaminants de l'environnement : lignées cellulaires et tests embryo-larvaires chez un poisson modèle le Médaka japonais, Oryzias latipes. Biodiversité et Ecologie. Université de Bordeaux, 2018. Français. ⟨NNT : 2018BORD0050⟩. ⟨tel-02139896⟩

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