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Ecologie De Legionella Pneumophila Dans Les Réseaux De Distribution D’Eau Potable Vincent Thomas

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Ecologie De Legionella Pneumophila Dans Les Réseaux De Distribution D’Eau Potable Vincent Thomas Ecologie de Legionella pneumophila dans les réseaux de distribution d’eau potable Vincent Thomas To cite this version: Vincent Thomas. Ecologie de Legionella pneumophila dans les réseaux de distribution d’eau potable. Ecologie, Environnement. Université Paris Sud - Paris XI, 2004. Français. tel-00116971 HAL Id: tel-00116971 https://tel.archives-ouvertes.fr/tel-00116971 Submitted on 29 Nov 2006 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. UNIVERSITÉ PARIS XI FACULTÉ DE PHARMACIE DE CHÂTENAY-MALABRY ÉCOLE DOCTORALE : INNOVATION THÉRAPEUTIQUE : DU FONDAMENTAL À L’APPLIQUÉ PÔLE : MICROBIOLOGIE / PARASITOLOGIE ANNÉE 2003-2004 SÉRIE DOCTORAT N°807 THÈSE Présentée À L’UNITÉ DE FORMATION ET DE RECHERCHE FACULTÉ DE PHARMACIE DE CHÂTENAY-MALABRY UNIVERSITÉ PARIS XI pour l’obtention du grade de DOCTEUR DE L’UNIVERSITÉ PARIS XI par Vincent THOMAS ECOLOGIE DE LEGIONELLA PNEUMOPHILA DANS LES RESEAUX DE DISTRIBUTION D’EAU POTABLE Soutenue le : 05 avril 2004 JURY : Mme Anne COLLIGNON, Professeur Mr Jérôme ETIENNE, Professeur (Rapporteur) Mr Yves LEVI, Professeur Mr Jean-François LORET, Docteur Mme Christine ROQUES, Docteur (Rapporteur) 1 Table des matières TABLE DES MATIERES TABLE DES MATIERES ..........................................................................................................2 REMERCIEMENTS………………………………………………………………………………….. 9 LISTE DES FIGURES ET TABLEAUX …………………………………………………………...12 LISTE DES ABREVIATIONS .................................................................................................16 CHAPITRE I : INTRODUCTION ………………………………………………………19 INTRODUCTION GENERALE ...............................................................................................20 I – LEGIONELLA PNEUMOPHILA : POINT SUR LES CONNAISSANCES ..........................25 I-1. BIOLOGIE ....................................................................................................................25 I-1.1. Taxonomie.............................................................................................................25 I-1.2. Caractères biologiques .........................................................................................27 I-1.3. Epidémiologie........................................................................................................27 I-1.4. Physiopathologie et clinique..................................................................................28 I-1.5. Facteurs de virulence de Legionella pneumophila................................................30 I-1.5.1 Le système Dot/Icm.........................................................................................31 I-1.5.2 Autres facteurs de virulence ............................................................................34 I-1.5.2.a. Structures de surface ................................................................................34 I-1.5.2.b. Sécrétion et acquisition de métabolites.....................................................36 I-1.5.3.c. Infection des macrophages .......................................................................37 I-1.5.3.d. Facteurs de virulence divers .....................................................................37 I-1.5.3. Transfert conjugatif de la virulence chez L. pneumophila ..............................38 I-1.6. Cycle intracellulaire de Legionella pneumophila ...................................................38 I-1.6.1. Evènements précoces ....................................................................................38 I-1.6.1.a. Phagocytose .............................................................................................38 I-1.6.1.b. Interception du trafic vésiculaire et inhibition de la fusion phagolyso........39 I-1.6.2. Réplication et fusion avec le réticulum endoplasmique..................................40 I-1.6.3. Sortie de la cellule ..........................................................................................43 I-1.7. Cycle intra-amibien de Legionella pneumophila....................................................43 I-1.8. Cycles intracellulaires (cellules mammifères et protozoaires) des autres espèces de légionelles .................................................................................................................44 I-2. DETECTION ET TYPAGE DES LEGIONELLES .........................................................48 I-2.1. Détection des légionelles ......................................................................................48 I-2.1.1. Culture ............................................................................................................48 I-2.1.2. Méthodes moléculaires...................................................................................50 I-2.1.2.a. Extraction de l'ADN et PCR.......................................................................50 I-2.1.2.b. Sondes ciblant l'ARN ribosomal ................................................................51 I-2.1.2.c. Autres méthodes .......................................................................................52 I-2.2. Diagnostic .............................................................................................................53 I-2.2.1. Culture ............................................................................................................53 I-2.2.2. Immunofluorescence directe ..........................................................................53 I-2.2.3. Détection des antigènes urinaires ..................................................................54 I-2.2.4. Diagnostic sérologique : Immunofluorescence indirecte ................................54 I-2.2.5. Amplification génique .....................................................................................55 I-2.3. Typage ..................................................................................................................55 I-2.3.1. Discrimination des espèces et des sous-groupes ..........................................56 I-2.3.1.a. Protéines totales et acides gras cellulaires ...............................................56 I-2.3.1.b. Polymorphisme des séquences des gènes mip, rpoB et dotA..................56 2 Table des matières I-2.3.1.c. Polymorphisme de longueur de l'espace intergénique de l'ADNt..............56 I-2.3.2. Discrimination des souches............................................................................57 I-2.3.2.a. Mobilité électrophorétique des enzymes...................................................57 I-2.3.2.b. Sérotypage................................................................................................57 I-2.3.2.c. ADN ribosomal ..........................................................................................57 I-2.3.2.d. Amplification aléatoire du chromosome : AP-PCR et RAPD.....................58 I-2.3.2.e. Polymorphisme de longueur des fragments amplifiés : AFLP & IRS-PCR59 I-2.3.2.f. Electrophorèse en champ pulsé (PFGE)....................................................59 I-2.3.2.g. Séquences intergéniques : variable-number tandem repeats...................60 I-2.3.2.h. Analyse de séquences ..............................................................................60 I-3. ECOLOGIE DE LEGIONELLA PNEUMOPHILA..........................................................60 I-3.1. Besoins nutritifs.....................................................................................................60 I-3.2. Présence dans l'environnement ............................................................................61 I-3.3. Interactions connues avec d'autres espèces bactériennes...................................62 I-3.3.1. Inhibition .........................................................................................................62 I-3.3.1.a. Inhibition par compétition ou bactéricidie ..................................................64 I-3.3.1.b. Parasitisme ...............................................................................................64 I-3.3.2. Potentialisation ...............................................................................................65 I-3.3.2.a. Bactéries ...................................................................................................65 I-3.3.2.b. Cyanobactéries .........................................................................................66 I-3.4. Interactions connues avec des espèces eucaryotes.............................................67 I-3.4.1. Amibes............................................................................................................69 I-3.4.2. Ciliés...............................................................................................................70 I-3.4.3. Dictyostelium discoideum ...............................................................................70 I-3.4.4. Algues vertes..................................................................................................71 I-3.5. Phages ..................................................................................................................71
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