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Réponse Transcriptomique Et Cellulaire De L'ormeau Rouge

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Réponse Transcriptomique Et Cellulaire De L'ormeau Rouge R´eponse transcriptomique et cellulaire de l'ormeau rouge Haliotis rufescens, cultiv´een ´ecloserieindustrielle face aux stress m´etalliqueset aux pathog`enes: r^oledes probiotiques dans la survie des organismes Fernando Silva Aciares To cite this version: Fernando Silva Aciares. R´eponse transcriptomique et cellulaire de l'ormeau rouge Haliotis rufescens, cultiv´een ´ecloserieindustrielle face aux stress m´etalliqueset aux pathog`enes: r^ole des probiotiques dans la survie des organismes. Zoologie des invert´ebr´es.Universit´ede Bretagne occidentale - Brest, 2013. Fran¸cais. <NNT : 2013BRES0058>. <tel-01089606> HAL Id: tel-01089606 https://tel.archives-ouvertes.fr/tel-01089606 Submitted on 2 Dec 2014 HAL is a multi-disciplinary open access L'archive ouverte pluridisciplinaire HAL, est archive for the deposit and dissemination of sci- destin´eeau d´ep^otet `ala diffusion de documents entific research documents, whether they are pub- scientifiques de niveau recherche, publi´esou non, lished or not. The documents may come from ´emanant des ´etablissements d'enseignement et de teaching and research institutions in France or recherche fran¸caisou ´etrangers,des laboratoires abroad, or from public or private research centers. publics ou priv´es. THÈSE / UNIVERSITÉ DE BRETAGNE OCCIDENTALE présentée par sous le sceau de l’Université européenne de Bretagne Fernando Silva Aciares pour obtenir le titre de DOCTEUR DE L’UNIVERSITÉ DE BRETAGNE OCCIDENTALE Préparée à L'institut Universitaire Européen Mention :Océanographie biologique de la Mer, Laboratoire des Sciences de École Doctorale Ecole Doctorale des Sciences de La Mer l'Environnement Marin (LEMAR) Thèse soutenue le 22 mars 2013 Devant le jury composé de : Réponse transcriptomique et Nathalie Cochennec Dr. HDR Cadre de recherche, IFREMER/ La Trinité Sur Mer/ Rapporteur cellulaire de l’ormeau rouge Haliotis Jean Marc Lebel rufescens , cultivé en écloserie Professeur, Université de Caen Basse Normandie/ Rapporteur Michel Auffret industrielle face aux stress Professeur, Université de Bretagne Occidentale/ Examinateur métalliques et aux pathogènes. Rôle Arnaud Tanguy des probiotiques dans la survie des Dr. MCF, Université de Paris 6 / Examinateur Carlos Riquelme organismes. Dr. Université d'Antofagasta (Chili)/ Co-directeur Dario Moraga Dr. MCF HDR, Université de Bretagne Occidentale/ Directeur THÈSE Présentée pour l’obtention du grade de Docteur de l’Université de Bretagne Occidentale Ecole Doctorale des Sciences de la Mer Océanographie Biologique Institut Universitaire Européen de la Mer Fernando Silva Aciares Réponse transcriptomique et cellulaire de l’ormeau rouge Haliotis rufescens , cultivé en écloserie industrielle face aux stress métalliques et aux pathogènes. Rôle des probiotiques dans la survie des organismes. Remerciements C’est avec le plus vif enthousiasme et le plus sincère que je voudrais remercier tous ceux qui à leur manière m’ont aidé à mener à bien cette thèse. Tout d’abord Je remercie grandement le Dr. Dario Moraga pour avoir assuré la direction de cette thèse. Merci pour la rigueur, le soutien, les conseils précieux, la disponibilité et en particulier pour son amitié. Merci de m’avoir fait confiance au cours de ces années de thèse. Je le remercie aussi pour tous les moments agréables passés à profiter des bons produits de la mer Bretonne. Je tiens à exprimer toute ma reconnaissance au Dr. Carlos Riquelme, mon co-directeur de thèse, pour la confiance qu’il m’a accordée en me permettant de présenter ces travaux dans le cadre d’une thèse d’Université. J’adresse un remerciement spécial au Dr. Michel Auffret pour son aide précieuse en immunologie, pour ses conseils et sa sympathie. Merci au Dr. Arnaud Tanguy pour ses conseils, sa disponibilité permanente et sa grande rigueur dans le domaine de la biologie moléculaire. Merci à mon cher ami Manuel Zapata « Zapatín » pour son amitié, ses conseils, sa disponibilité et l’aide qu’il a su m’apporter dans les manips au cours de cette thèse. Je tiens à remercier à Jean Luc et Véronique Queau pour leur amitié et leur accueil. Je tiens à remercier Sleiman Bassin pour son aide dans les manips et les bonnes remarques. Merci également à Camille Lacroix et Alain Marhic pour leur aide précieuse. Je tiens à remercier chaleureusement Brigitte Moraga (Pôle langues de l’Université de Bretagne Occidentale) pour la révision finale de ce manuscrit. Je voudrais aussi remercier tous les membres du jury qui ont accepté d’examiner ce travail. Mes remerciements s’adressent particulièrement à ma famille, pour son appui, sa confiance et son amour qui m’ont été si précieux durant toute ma vie. SOMMAIRE LISTE DES TABLEAUX ET FIGURES LISTE DES ABREVIATIONS INTRODUCTION GENERALE…………………………………………………….. 1 1. Stress environnemental: Implications des métaux traces et des pathogènes…….…...… 2 2. Contexte de l’étude et objectifs…………………...……………………………………..6 RAPPELS BIBLIOGRAPHIQUES……………………………………………….…………………9 1. Présentation générale du modèle biologique…………………..…………………………...10 1.1. Morphologie……...….……………………………………………………............11 1.2. Reproduction et développement…………………………………………………..14 1.3. Alimentation………………………………………………………………………15 1.4. Habitat…………………………………………………………………….……….16 1.5. Importance en aquaculture…………………………..………………………….…16 1.6. Cycle de culture et zootechnie…………………………………………………… 18 2. Généralités sur le multi stress dans la culture de l’ormeau: Cas particulier du cuivre.. 21 2.1. L’impact du cuivre sur les mollusques marins……………………………………24 3. Généralités sur le multi stress dans la culture de l’ormeau: Cas particulier des micro- organismes pathogènes ( Vibrionaceae )………………………………………………..……. 28 3.1. Les vibrions pathogènes…………………………………………………………..32 3.1.1. Chez les mollusques…………………………………………….……….32 3.1.1.1. Impact des pathogènes chez l’ormeau………………….……...34 3.1.2. Vibrio parahaemolyticus ………………………………………….……...38 3.1.2.1. Généralités……………………………………………….…….38 3.1.2.2. Habitat et cycle de vie…………………………………….…...41 3.1.2.3. Les gènes de virulence et les régulateurs………….…………..42 3.1.2.4. Contrôle du quorum sensing (QS)………………….…………44 3.1.2.5. Effet de V. parahaemolyticus chez l’ormeau……….………....46 4. Rôle des probiotiques dans les activités aquacoles. Généralités………..……………... 48 4.1. Définition des micro-organismes probiotiques…………………………………...48 4.2. Utilisation des micro-organismes probiotiques pour la prévention des maladies..50 4.2.1. Application des probiotiques chez l’ormeau H. rufescens ……….……..53 4.3. Critères de sélection………………………………………………………………53 4.4. Voies d'incorporation……………………………………………………………..54 4.5. Possible mode d'action……………………………………………………………54 4.5.1. Production de composés antibactériens…………………………………55 4.5.2. Compétition pour les nutriments et sites de fixation chez l´hôte………57 4.5.3. Quorum quenching………………………………………………………58 4.5.4. Stimulation du système immunitaire……………………………………60 4.5.5. Amélioration de la qualité de l'eau……………………………………...60 5. Généralités sur les mécanismes immunitaires des mollusques marins……………… 61 6. Généralités sur l’expression génique chez l'ormeau H. rufescens …………………... 69 CHAPITRE I : Effet des probiotiques sur la survie et la croissance chez les juvéniles de l’ormeau rouge H. rufescens cultivés en écloserie industrielle…………………………….……… 70 Contexte de l’étude……………………………………………………………………………………71 Article N°1 : Silva-Aciares, F ., Mejías, C., Carvajal, P., Riquelme, C., 2011 a. Use of macroalgae supplemented with probiotics in the Haliotis rufescens culture in the northern Chile. Aquaculture Research 42 : 953-961……………………………………………………………………………….. 75 Conclusion sur le rôle des probiotiques……………………………………………………………….84 CHAPITRE II : Caractérisation de la réponse moléculaire et cellulaire chez les juvéniles de Haliotis rufescens face au cuivre en conditions expérimentales…………………….…………….. 87 Contexte de l’étude…………………………………………………………………………………….88 A. Identification des gènes exprimés chez les Ormeaux juvéniles d’ Haliotis rufescens en réponse à différentes concentrations de cuivre en conditions contrôlées………………………………………...92 Généralités……………………………………………………………………………………..93 • Article N°2: Silva-Aciares, F ., Zapata, M., Tournois, J., Moraga, D., Riquelme, C., 2011 b. Identification of genes expressed in juvenile Haliotis rufescens in response to different copper concentrations in the north of Chile under controlled conditions. Marine Pollution Bulletine 62: 2671-2680………………………………………….. 94 • Conclusion sur le chapitre II A………………………………………………………104 B. Effets du cuivre sur les activités d’immunomodulation hémocytaire d’Haliotis rufescens ……...105 • Généralités……………………………………………………………………………106 • Article N°3: Silva-Aciares, F ., Moraga, D., Riquelme, C., 2013 . Effect of copper on the immunomodulatory activities of hemocytes in juvenile abalone, Haliotis rufescens cultivate in hatchery conditions. Soumis à Aquaculture…………………………… 108 • Conclusion sur le chapitre II B………………………………………………………129 CHAPITRE III : Réponse genomique et cellulaire de H. rufescens nourris avec des probiotiques et cultivé en écloserie industrielle face aux stress des micro-organismes pathogènes………….. 131 • Contexte et introduction……………………………………….……………………..132 • Article N°4: Silva-Aciares, F ., Moraga D., Auffret, M., Tanguy, A., Riquelme, C., 2013 . Transcriptomic and cellular response to bacterial challenge (Pathogenic Vibrio parahaemolyticus ) in farmed juvenile Haliotis rufescens fed with or without probiotic diet. Journal of Invertebrate Pathology 113:163-176 ……..………………………137 • Conclusion……………………………………………………………………………155 CONCLUSIONS GENERALES ET PERSPECTIVES
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