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Ecophysiological Studies on Dinophysis and Its Food Chain, And THESE DE DOCTORAT DE L'UNIVERSITE DE NANTES ECOLE DOCTORALE N° 598 Sciences de la Mer et du littoral Spécialité : Biologie marine Par Sylvain GAILLARD Études écophysiologiques sur Dinophysis et sa chaîne alimentaire, et effets in vitro du dinoflagellé et de ses toxines sur les premiers stades de vie de deux modèles d'animaux marins (l’huître et le poisson) “Ecophysiological studies on Dinophysis and its food chain, and in vitro effects of the dinoflagellate and its toxins on early life stages of two models of marine animals (oyster and fish)” Thèse présentée et soutenue à Nantes, le 27 novembre 2020 Unité de recherche : Laboratoire Phycotoxines (PHYC), IFREMER Centre Atlantique de Nantes Rapporteurs avant soutenance : Beatriz REGUERA Directrice de recherche, Instituto Español de Oceanografia - Centro Oceanográfico de Vigo Per Juel HANSEN Professeur des universités, University of Copenhagen - Department of Biology Composition du Jury : Rapporteure : Beatriz REGUERA Directrice de recherche, Instituto Español de Oceanografia - Centro Oceanográfico de Vigo Rapporteur : Per Juel HANSEN Professeur des universités, University of Copenhagen - Department of Biology Président : Laurent BARILLÉ Professeur des universités, Université de Nantes - MMS Examinatrice : Claudia WIEGAND Professeure des universités, Université de Rennes 1 - CNRS - ECOBIO Directeur de thèse : Philipp HESS Docteur, Cadre de recherche, Laboratoire Phycotoxines, IFREMER Nantes Co-directrice de thèse : Hélène HÉGARET Docteure, Chargée de recherche, CNRS - LEMAR, Plouzané Invités : Véronique SÉCHET Docteure, Cadre de recherche, Laboratoire Phycotoxines, IFREMER Nantes Damien RÉVEILLON Docteur, Cadre de recherche, Laboratoire Phycotoxines, IFREMER Nantes Wolfgang VOGELBEIN Directeur de recherche, Virginia Institute of Marine Science, USA Acknowledgements I first would like to gratefully acknowledge the members of my committee and the examiners of my PhD thesis, Beatriz Reguera, Per Juel Hansen, Claudia Wiegand and Laurent Barillé. Ma thèse s’est déroulée au laboratoire Phycotoxines de l’IFREMER de Nantes, je remercie donc Zouher Amzil son directeur, et a été financée grâce au projet CoCliME. Merci à mes deux directeurs de thèse, Philipp Hess et Hélène Hégaret ainsi qu’à mes deux co- encadrants, Véronique Séchet et Damien Réveillon, pour m’avoir permis de réaliser cette thèse dans les meilleures conditions possibles, pour m’avoir fait confiance, pour votre aide et vos encouragements. Pour ces raisons, et pour tout le reste je vous serai toujours reconnaissant. I would like to extend my gratitude to Wolfgang Vogelbein for welcoming me to your laboratory in Virginia and for teaching me a few of your knowledge. Je remercie les membres de mon comité de thèse, Rodolphe Lemée, Thomas Lacour, Manoella Sibat et Marc Sourisseau pour pour vos conseils et encouragements. Merci à tous mes collègues de Phyc, Mano, Fabienne, Michelle, Véro, Liliane, Charline, Énora, Georges et Korian pour tous ces moments partagés, au travail et en dehors. Un grand merci à mes collaborateurs au labo, mais aussi à PBA, Aurélie, Thomas et Gaël, et au LEMAR à Brest, Nelly, Caroline, Myrina, Justine, Lucas, Marc, pour votre aide pendant les manips et pour la rédaction. Merci Nour, Solène, Marin, Thomas, Seb, Florent, Élise, Estelle, Cyrielle, François et Max on aura bien rigolé ! Également, merci à ma famille et mes ami(e)s. 3 Table of contents Acknowledgements .......................................................................................................................... 3 Table of contents .............................................................................................................................. 4 List of figures ................................................................................................................................. 11 List of tables ................................................................................................................................... 16 List of abbreviations ....................................................................................................................... 21 Contexte et objectifs de la thèse ..................................................................................................... 24 Context and aims of the thesis ........................................................................................................ 34 Chapter 1: State of the art - Literature review ................................................................................ 43 I-1) General characteristics of the dinoflagellates ................................................................................... 44 I-2) Morphology and taxonomy of Dinophysis ....................................................................................... 44 I-3) Dinophysis food chain ...................................................................................................................... 47 I-3-1) First tier of the food chain: T. amphioxeia ................................................................................ 47 I-3-2) Mixotrophy of M. rubrum ......................................................................................................... 48 I-3-3) Mixotrophy of Dinophysis ........................................................................................................ 50 I-4) Culture of the three-tier food chain of Dinophysis. .......................................................................... 52 I-4-1) Culture of T. amphioxeia .......................................................................................................... 53 I-4-2) Culture of M. rubrum ................................................................................................................ 54 I-4-3) Culture of Dinophysis ............................................................................................................... 55 I-5) Ecophysiological experiments with Dinophysis species .................................................................. 57 I-5-1) Temperature .............................................................................................................................. 57 I-5-2) Irradiance .................................................................................................................................. 59 I-5-3) Salinity ...................................................................................................................................... 62 I-5-4) Nutrients .................................................................................................................................... 62 I-5-5) Geographical origin ................................................................................................................... 64 I-5-6) Prey ........................................................................................................................................... 65 I-5-7) Growth phase and cell cycle ..................................................................................................... 66 I-5-8) Conclusion of the ecophysiological experiments on Dinophysis .............................................. 67 I-6) Dinophysis Toxins ............................................................................................................................ 68 4 I-6-1) OA-group toxins ....................................................................................................................... 68 I-6-1-1) Chemical structure of OA-group toxins ............................................................................. 68 I-6-1-2) Toxicity of OA-group toxins by intraperitoneal injection and oral administration in rodents ............................................................................................................................................. 69 I-6-1-3) Elucidation of action mode of OA-group toxins................................................................ 70 I-6-2) PTXs ......................................................................................................................................... 71 I-6-2-1) Chemical structure of PTXs ............................................................................................... 71 I-6-2-2) Toxicity of PTXs by intraperitoneal injection and oral administration in rodents ............ 72 I-6-2-3) Elucidation of action mode of PTXs .................................................................................. 73 I-6-3) Effects of OA-group toxins and PTXs on humans .................................................................... 74 I-6-4) OA-group toxin and PTX measurements .................................................................................. 74 I-6-4-1) Functional methods ............................................................................................................ 75 I-6-4-2) Immunological methods .................................................................................................... 75 I-6-4-3) Chemical analytical methods ............................................................................................. 75 I-7) Accumulation of Dinophysis, OA-group toxins and PTXs in marine organisms ............................. 76 I-7-1) Accumulation in crustaceans, echinoderms, gastropods and cephalopods ............................... 76 I-7-2) Accumulation in filter-feeding bivalves ...................................................................................
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