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Moustiques Et Dirofilariose: Mise Au Point Et Utilisation D'outils Innovants

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Moustiques Et Dirofilariose: Mise Au Point Et Utilisation D'outils Innovants AVANT PROPOS Le format de présentation de cette thèse correspond à une recommandation de la spécialité Maladies Infectieuses et Microbiologie, à l’intérieur du Master des Sciences de la Vie et de la Santé qui dépend de l’Ecole Doctorale des Sciences de la Vie de Marseille. Le candidat est amené à respecter des règles qui lui sont imposées et qui comportent un format de thèse utilisé dans le Nord de l’Europe et qui permet un meilleur rangement que les thèses traditionnelles. Par ailleurs, la partie introduction et bibliographie est remplacée par une revue envoyée dans un journal afin de permettre une évaluation extérieure de la qualité de la revue et de permettre à l’étudiant de commencer le plus tôt possible une bibliographie exhaustive sur le domaine de cette thèse. Par ailleurs, la thèse est présentée sur article publié, accepté ou soumis, associé d’un bref commentaire donnant le sens général du travail. Cette forme de présentation a paru plus en adéquation avec les exigences de la compétition internationale et permet de se concentrer sur des travaux qui bénéficieront d’une diffusion internationale. Pr Didier RAOULT REMERCIEMENTS Au terme de ce travail de thèse, je tiens à remercier tout particulièrement : Monsieur le Pr Philippe PAROLA, mon directeur de thèse. Son encadrement, sa rigueur scientifique et sa disponibilité m’ont permis de réaliser ce travail. Pour les efforts qu’il a consentis lors de la correction de mes manuscrits d’articles. Il m’a fait découvrir d’autres horizons scientifiques au cours de ma formation doctorale. Je le prie d’accepter l’expression de mes sincères remerciements. Monsieur le Dr vétérinaire Bernard DAVOUST pour avoir co-encadré ce travail. Sa disponibilité, son soutien et ses conseils m’ont permis de mener à bien mes travaux de recherche. Je lui suis reconnaissant pour la confiance qu'il m'a accordée et j’espère avoir été à la hauteur de ses attentes. Qu’il trouve ici l’expression de ma profonde gratitude. Monsieur Lionel ALMERAS pour le soutien qu’il m’a apporté tout au long de cette thèse. Pour le suivi constant de mes travaux et sa contribution scientifique. Ce travail de thèse a été réalisé en collaboration avec CEVA Santé Animale. Aussi, je remercie, vivement, Madame Marie VARLOUD, responsable technique Parasitologie pour les animaux de compagnie, Monsieur le Dr vétérinaire Gautier BARTHELEMY directeur de l’évaluation des projets pour les animaux de compagnie et Monsieur le Dr vétérinaire Hamadi KAREMBE, directeur de l’innovation au département R&D Pharmacologie. Votre appui scientifique ainsi que votre soutien moral et financier ont été déterminants dans la réussite de ce travail. Je vous prie d’accepter tous, ainsi que la Direction de CEVA, l’expression de mes sincères remerciements. Mes remerciements s’adressent aux membres de mon jury : Madame le Pr Florence FENOLLAR, qui me fait le très grand honneur de présider le jury de cette thèse. Qu'elle trouve ici l'expression de ma parfaite reconnaissance et mes vifs remerciements. Monsieur le Dr Arezki IZRI pour avoir accepté de juger mon travail. Sincères remerciements. Monsieur Fabrice CHANDRE qui a accepté d’évaluer mon travail. Veuillez trouver ici l’expression de ma profonde reconnaissance. Monsieur le Dr vétérinaire Radu BLAGA pour avoir bien voulu examiner mon travail. Sincères remerciements. Mes sincères remerciements vont aussi à Monsieur le Pr Didier RAOULT, directeur de l’URMITE, pour m’avoir accueilli au sein de son laboratoire. Je remercie chaleureusement l’ensemble de mes collègues pour leur soutien et leurs encouragements dans l’achèvement de ce travail. Je ne saurais terminer mes propos sans adresser un grand merci à tous ceux qui ont contribué, de près ou de loin, à la réussite de ce travail, tout au long des trois dernières années. DÉDICACES A MES TRES CHERS PARENTS pour leurs encouragements et leur soutien permanent. A MES FRÈRES & SŒURS pour tous les bons moments passés ensemble et pour tous ceux à venir… pour leur soutien et leur aide. SOMMAIRE RÉSUMÉ/ABSTRACT 2/4 I – INTRODUCTION 7 Article N° 1 : Revue de la littérature. Tahir D, Davoust B and 17 Parola P. Vector-borne helminth diseases in pets and humans in the Mediterranean Basin: an update. II- MISE AU POINT D’UN OUTIL MOLÉCULAIRE 59 POUR LA DETECTION DE DIROFILARIA SPP. Article N° 2 : Tahir D, Bittar F, Barré-Cardi H, Mediannikov O, Raoult D, Davoust B and Parola P. Molecular survey of 63 Dirofilaria immitis and D. repens by Taqman real-time PCR in dogs and mosquitoes (Diptera: Culicidae) in Corsica. Vet. Parasitol. 2017, 235:1–7. Article N° 3 : Tahir D, Damene H, Davoust B and Parola P. First molecular detection of Dirofilaria immitis (Spirurida: 73 Onchocercidae) infection in dogs from Northern Algeria. Comp. Immunol. Microbiol. Infect. Dis. 2017, 51:66-68. III- APPLICATION DU MALDI-TOF MS POUR LA DETECTION DES FILAIRES DANS LES MOUSTIQUES 79 Article N° 4 : Tahir D, Almeras L, Varloud M, Raoult D, Davoust B and Parola P. Assessment of MALDI TOF mass spectrometry for filariae detection in Aedes aegypti mosquitoes. 83 Accepted to PloS Neglected Tropical Diseases: Vector-borne diseases. IV- EVALUATION DES INSECTICIDES CONTRE LES 123 VECTEURS Article N° 5 : Tahir D, Davoust B, Almeras L, Berenger JM, Varloud M and Parola P. Anti-feeding and insecticidal efficacy of a topical administration of dinotefuran-pyriproxyfen- 129 permethrin spot-on (Vectra® 3D) on mice against Stegomyia albopicta (= Aedes albopictus). Med. Vet. Entomol. 2017, 31(4):. Article N° 6 : Tahir D, Davoust B, M. Varloud M, Berenger JM, Raoult D, Almeras L and Parola P. Assessment of the anti-feeding 139 and insecticidal effects of the combination of dinotefuran, permethrin and pyriproxyfen (Vectra® 3D) against Triatoma infestans on rats. Med. Vet. Entomol. 2017, 31(2):132-139. IV- CONCLUSION GENERALE ET PERSPECTIVE 149 REFERENCES 153 ANNEXES 159 Article N° 7 : Tahir D, Alwassouf S, Loudahi A, Davoust B and Charrel RN. Seroprevalence of Toscana virus in dogs from 163 Kabylia (Algeria). Clin. Microbiol. Infect. 2016, 22/e16-e17. Article N° 8 : Tahir D, Socolovschi C, Marié JL, Ganay G, Berenger JM, Bompar JM, Blanchet D, Cheuret M, Mediannikov O, Davoust B and Parola P. New Rickettsia species in soft ticks 167 collected from bats in French Guiana. Ticks Tick Borne Dis. 2016, 7(6):1089-1096. Article N° 9 : Dahmani M, Davoust B, Tahir D, Fenollar F, and Mediannikov O. Molecular investigation and phylogeny of 177 Anaplasmataceae species infecting domestic animals and ticks in Corsica, France. Parasit. Vectors. 2017, 10(1):302. Article N° 10 : Tahir D, Davoust B, Heu K, Lamour T, Marié JL and Blanchet D. Molecular and serological investigation of 191 Trypanosoma cruzi infection in dogs in French Guiana. Vet. Parasitol. Reg. Stud. Rep. 2017. RÉSUMÉ Les moustiques et les tiques sont les principaux vecteurs incriminés dans la transmission d’agents pathogènes aux carnivores et à l’homme. Les dirofilarioses sont des infections parasitaires transmises par différentes espèces de moustiques. Ces maladies parasitaires sont imputables à deux espèces de filaires : Dirofilaria immitis et Dirofilaria repens adaptées aux hôtes canins, félins et humains. Dans ce travail, nous nous sommes intéressés à l’étude des dirofilarioses chez le réservoir canin « le chien » ainsi que chez les vecteurs « moustiques », en particulier, en ce qui concerne la détection, la surveillance et la prophylaxie. Le premier objectif était de développer une PCR duplex en temps réel ciblant le gène COI capable de détecter et de différencier simultanément D. immitis et D. repens à partir d'échantillons de moustiques ou de sang périphérique chez les carnivores. Par la suite, nous avons appliqué cet outil pour la surveillance de la dirofilariose canine dans différentes zones enzootiques du bassin méditerranéen (Corse et Algérie). Ainsi, nous avons détecté par cet outil moléculaire, pour la première fois en France, D. immitis et D. repens chez des moustiques tigre « Aedes albopictus (ou Stegomyia albopicta) » collectés en Corse. Nous avons, de plus, confirmé la présence de l’infection à D. immitis chez les chiens du nord de l'Algérie. 2 Le deuxième objectif était d'évaluer l’intérêt de la spectrométrie de masse, MALDI-TOF MS, pour la détection de changements dans les profils protéiques d'Aedes aegypti infectés expérimentalement avec des nématodes filaires (D. immitis, Brugia malayi et B. pahangi) par rapport à ceux qui ne sont pas infectés, et ce, en testant différentes parties des moustiques. Les résultats obtenus montrent la capacité du MALDI-TOF MS à différencier des moustiques infectés et non infectés par les filaires. Ainsi, les meilleurs taux de classification correcte ont été obtenus à partir de la partie « tête-thorax » du moustique avec 94,1%, 86,6%, 71,4% et 68,7% pour les non infectés versus ceux infectés, respectivement, par D. immitis, B. malayi et B. pahangi. Le troisième objectif de ce travail était l’évaluation de l'efficacité anti-gorgement et insecticide d'un ectoparasiticide (Vectra® 3D) contenant trois principes actifs : le dinotéfurane, le pyriproxyfène et la perméthrine (DPP) contre Ae. albopictus, l'une des principales espèces vectrices de Dirofilaria spp. ceci en utilisant un modèle murin. Les résultats ont démontré que le DPP a une efficacité anti-gorgement et insecticide significative contre Ae. albopictus pendant au moins un mois, suggérant que cette combinaison (DPP) aurait une bonne efficacité contre un des vecteurs de la dirofilariose canine. Mots-clés : Moustiques, Dirofilaria spp., PCR en temps réel, MALDI-TOF MS, ectoparasiticide et surveillance. 3 ABSTRACT Mosquitoes and ticks are the main vectors of pathogen transmission to carnivores and humans. Dirofilarial diseases are mosquito-borne parasitic infections caused by two main filarial species (Dirofilaria immitis and Dirofilaria repens) which have adapted to canine, feline, and human hosts.
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