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Etude De La Pollution Marine Par Les Hydrocarbures Et Caractérisation De Leurs Effets Biochimiques Et Moléculaires Sur La Palourde De Ruditapes Sp

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Etude De La Pollution Marine Par Les Hydrocarbures Et Caractérisation De Leurs Effets Biochimiques Et Moléculaires Sur La Palourde De Ruditapes Sp Etude de la pollution marine par les hydrocarbures et caractérisation de leurs effets biochimiques et moléculaires sur la palourde de Ruditapes sp. Houssem Chalghmi To cite this version: Houssem Chalghmi. Etude de la pollution marine par les hydrocarbures et caractérisation de leurs effets biochimiques et moléculaires sur la palourde de Ruditapes sp.. Géochimie. Université deBor- deaux, 2015. Français. NNT : 2015BORD0175. tel-01583022 HAL Id: tel-01583022 https://tel.archives-ouvertes.fr/tel-01583022 Submitted on 6 Sep 2017 HAL is a multi-disciplinary open access L’archive ouverte pluridisciplinaire HAL, est archive for the deposit and dissemination of sci- destinée au dépôt et à la diffusion de documents entific research documents, whether they are pub- scientifiques de niveau recherche, publiés ou non, lished or not. The documents may come from émanant des établissements d’enseignement et de teaching and research institutions in France or recherche français ou étrangers, des laboratoires abroad, or from public or private research centers. publics ou privés. Logo Université de cotutelle THÈSE EN COTUTELLE PRÉSENTÉE POUR OBTENIR LE GRADE DE DOCTEUR DE L’UNIVERSITÉ DE BORDEAUX ET DE L’UNIVERSITÉ DE MONASTIR ÉCOLE DOCTORALE SCIENCES ET ENVIRONNEMENTS ÉCOLE DOCTORALE SCIENCES BIOLOGIQUES, BIOTECHNOLOGIE ET SANTÉ SPÉCIALITÉ : GÉOCHIMIE ET ÉCOTOXICOLOGIE Par Houssem CHALGHMI ÉTUDE DE LA POLLUTION MARINE PAR LES HYDROCARBURES ET CARACTÉRISATION DE LEURS EFFETS BIOCHIMIQUES ET MOLÉCULAIRES SUR LA PALOURDE RUDITAPES SP. Sous la direction de : Jean-Paul BOURDINEAUD et Dalila SAIDANE-MOSBAHI Soutenue le 06 octobre 2015 Membres du jury : M. BOUSSETTA, Hamadi Professeur, Université de Sousse Président Mme MOUNEYRAC, Catherine Professeur, Université Catholique de l’Ouest Rapporteur M. BANNI, Mohamed Professeur, Université de Monastir Rapporteur Mme GIAMBÉRINI, Laure Professeur, Université de Lorraine Examinatrice M. BOURDINEAUD, Jean-Paul Professeur, Université de Bordeaux Directeur de thèse Mme SAIDANE-MOSBAHI, Dalila Professeur, Université de Monastir Directrice de thèse Titre : Étude de la pollution marine par les hydrocarbures et caractérisation de leurs effets biochimiques et moléculaires sur la palourde Ruditapes sp. Résumé : Dans notre étude, la biosurveillance de la lagune de Tunis a été réalisée durant une année en employant une approche combinant les analyses chimiques et biologiques et en utilisant la palourde Ruditapes decussatus comme espèce bioindicatrice. Cette approche a permis de mettre en évidence la forte contamination de la lagune de Tunis par les métaux traces et les hydrocarbures aromatiques polycycliques (HAPs). La variation saisonnière des niveaux de bioaccumulation de ces contaminant par la palourde s’avère être fortement associée aux changements des paramètres physico-chimiques du milieu et aux processus physiologiques de l’animal. L’étude de la réponse biologique basée sur l’utilisation d’une batterie de biomarqueurs d’exposition et d’effet situés à différents niveaux de l’organisation biologique : moléculaire (l’expression de cinq gènes d’intérêt), biochimique (les activités benzo(a)pyrène hydroxylase, glutathion S-transférase, acétylcholinestérase, catalase et taux de malondialdéhyde) et tissulaire (altérations histopathologiques), a permis de mettre en évidence une modulation des processus de défense contre les perturbations induites par la pollution et l’identification des altérations histopathologiques (structurales) au niveau des branchies et de la glande digestive impliquant un impact sévère des contaminants sur l’état de santé de la palourde. L’étude des interactions spatio-temporelles entre les facteurs abiotiques et biotiques a permis d’identifier la température et la reproduction comme paramètres principaux affectant la réponse biochimique de défense et d’effet. L’analyse en composante principale (ACP), regroupant tous les paramètres analysés pendant le printemps, nous a permis d’identifié le site Z2 comme le site le plus affecté par la pollution. Dans un second temps, le benzo(a)pyrène (BaP), déjà identifié dans la lagune de Tunis, a été employé dans des conditions contrôlées au laboratoire afin de caractériser les réponses moléculaires et biochimiques chez les bivalves (Ruditapes philippinarum et Crassostrea gigas) face à des expositions subaiguës et aiguës à ce contaminant. Cette étude nous a permis d’identifier une modulation des biomarqueurs biochimiques de métabolisation, de stress oxydant et de neurotoxicité et de l’expression des gènes impliqués dans les processus de métabolisation, de respiration mitochondriale, de défense antioxydante, de reproduction et de défense immunitaire. L’analyse de l’altération de l’ADN a révélé un pouvoir génotoxique précoce et élevé du BaP chez les deux bivalves. Mots-clés : Lagune de Tunis, bivalves, HAPs, métaux, benzo(a)pyrène, marqueurs biochimiques, histologie, réponse transcriptomique, génotoxicité. Title : Study of the marine pollution by hydrocarbons and characterization of their biochemical and molecular effects on clam Ruditapes sp. Abstract : Firstly, biomonitoring of Tunis lagoon was conducted during one year using an approach combining chemicals and biological analyses and the bioindicator species Ruditapes decussatus. This approach highlighted the high contamination of the Tunis lagoon by trace metals and polycyclic aromatic hydrocarbons (PAHs). Seasonal variation of contaminant bioaccumulation levels in clam was found to be strongly associated with physicochemical changes in surrounding environment and physiological processes in organism. Biological responses investigated through a battery of exposure and effect biomarkers located at different levels of biological organisation: molecular (expression of five genes), biochemical (benzo(a)pyrene hydroxylase, glutathione S-transferase, acetylcholinesterase and catalase activities and malondialdehyde concentration) and tissular (histopathological damages), demonstrated a defense process modulation by pollution and showed histopathological alterations in gills and digestive gland involving severe impact of contaminants on health state of clam. The study of spatiotemporal interactions between abiotic and biotic factors identified temperature and reproduction as main parameters affecting defense and effect biochemical response. The principal component analysis (PCA), using all analyzed parameters during the spring, allowed to identify site Z2 as the most affected by pollution. Secondly, benzo(a)pyrene (BaP), already identified in Tunis lagoon, was used in controlled conditions at the laboratory in order to characterize the molecular and biochemical responses in bivalves (Ruditapes philippinarum and Crassostrea gigas) facing a subacute and acute exposures to this contaminant. This study showed a modulation of biochemical biomarkers of metabolism, oxidative stress and neurotoxicity and expression of genes involved in process of metabolisation, mitochondrial respiration, antioxidant defense, reproduction and immune defense. The analysis of DNA damages revealed a high and early genotoxicity effect of BaP in both bivalves. Keywords: Tunis lagoon, bivalves, PAHs, metals, benzo(a)pyrene, biochemical biomarkers, histology, transcriptomic response, génotoxicity. UMR CNRS 5805 EPOC, Équipe Écotoxicologie Aquatique, Station Marine d’Arcachon, Place du Dr Peyneau, 33120 Arcachon, France. Laboratoire d’Analyse, Traitement et Valorisation des Polluants de l’Environnement et des Produits, Faculté de Pharmacie, Université de Monastir, 5000 Monastir, Tunisie. Dédicaces A mes très chers parents Hatem et Nourhène Qui par leur bonne volonté, leur patience, leurs conseils pertinents, leur assistance, m’ont aidé et m’ont soutenue tant sur le plan moral que financier pour voir se réaliser un de leurs vœux les plus chers. Sans leur soutien permanent cette thèse n’aurait pas pu voir le jour. J’aimerais vous remercier pour votre confiance en moi, votre encouragement, pour tous les efforts et les sacrifices que vous avez faits pour que je devienne ce que je suis. Puisse Dieu vous préserver en bonne santé et vous accorder bonheur et longue vie. A ma chère fiancée Siwar Pour ton amour, soutien, disponibilité et ta compréhension. Merci de m’avoir aidé dans les moments difficiles, tes encouragements m’ont été d’une grande rescousse. L’ampleur de tes sacrifices n’a d’égal que la grandeur de tes sentiments. Puisse ce travail te témoigner mon immense gratitude et surtout mon éternel amour. A mes chers frères Zied, Mahmoud et Achraf Pour votre encouragement et votre présence morale. Je vous souhaite un grand avenir et une vie prospère et rayonnante. A la mémoire de mes grands pères A toute la famille CHALGHMI et BEN KAHLA A ma belle famille SANDID Aimablement Houssem Remerciements Ce travail a été réalisé en cotutelle internationale entre l‘Institut Supérieur de Biotechnologie de Monastir (Université de Monastir) et l‘Université de Bordeaux au sein du Laboratoire d’Analyse, Traitement et Valorisation des Polluants de l’Environnement et des Produits à la Faculté de Pharmacie de Monastir et de l’UMR CNRS 5805 EPOC-Station Marine d’Arcachon. J’exprime toute ma gratitude à ma directrice de thèse, Madame le Professeur Dalila Saidane-Mosbahi, je vous remercie de m’avoir proposé un sujet de Master, de l’avoir transformé en sujet de thèse et pour avoir encadré mes travaux jusqu’à leurs aboutissements, pour m’avoir fait partager sa passion pour la recherche, son ouverture d’esprit et sa rigueur scientifique durant toutes ces années passées à
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