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Trombidiformes), Parasitos De Répteis E Anfíbios, No Estado De São Paulo

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Trombidiformes), Parasitos De Répteis E Anfíbios, No Estado De São Paulo JAIRO ALFONSO MENDOZA ROLDAN Estudos morfológicos e investigação da presença de bactérias e protozoários em ácaros (Trombidiformes), parasitos de répteis e anfíbios, no estado de São Paulo São Paulo 2015 FOLHA DE AVALIAÇÃO Nome: MENDOZA-ROLDAN, Jairo Alfonso Título: Estudos morfológicos e investigação da presença de bactérias e protozoários em ácaros (Trombidiformes), parasitos de répteis e anfíbios, no estado de São Paulo Dissertação apresentada ao Programa de Pós- Graduação em Epidemiologia Experimental Aplicada às Zoonoses da Faculdade de Medicina Veterinária e Zootecnia da Universidade de São Paulo, para obtenção do título de Mestre em Ciências Data: ____/____/______ Banca Examinadora Prof Dr._________________________________________________________ Instituição:__________________________Julgamento___________________ Prof Dr._________________________________________________________ Instituição:__________________________Julgamento___________________ Prof Dr._________________________________________________________ Instituição:__________________________Julgamento___________________ Dedicado a mi madre Luz Mery, Por tu apoyo incondicional y tus palabras siempre llenas de sabiduría. AGRADECIMENTOS Agradeço primeiramente a minha orientadora Dra. Darci Moraes Barros Battesti, que sempre acreditou em mim, que sem importar a situação sempre foi um apoio incondicional e cujas palavras e conselhos me ajudaram a crescer como profissional e como pessoa. Aos colegas do laboratório de Biologia Celular (Instituto Butantan), Dra. Marta Antoniazzi, Dr. Carlos Jared e em especial Beatriz Mauricio, pela valiosa ajuda na obtenção das imagens de MEV, pela amizade e bons momentos. A todos os colegas e amigos do laboratório de Parasitologia (Instituto Butantan), em especial Valeria Castilho, Simone Simons, Felipe Gomes, Natalia Isola, Polianni Carmozini, Taisy Mazur e Leonor que são como minha família. Muito obrigado! A Juliana Cuoco Badari, e sua família por me acolher como um filho no Brasil. Obrigado por me ensinar tanto e pela amizade. A Lívia Corrêa, e sua família por permitir conhecer parte deste país e por estar comigo nos momentos bons e não tão bons; A Mercedes Reyes por ser como minha mãe, pelas palavras e pelo valioso conhecimento em acarologia que me transmitiu. A todos os colegas, amigos e pesquisadores do Laboratório Especial de Coleções Zoológicas (Instituto Butantan) em especial a Leidiane Duarte, Maria Cristina Rosário. Gabi Andrade, Diego Ramirez, Gabriel Alves, Dalton Giovanni, Ricardo Silva, Fernando Jacinavicius, Marcia Fugiwara, Daniella Franze e Angelina Cirelli Moraes, por sua amizade incondicional, pelos conselhos e pela ajuda em cada passo do desenvolvimento deste projeto. Ao diretor Roberto H. Pinto Moraes, por permitir que concluísse o mestrado. A Fernanda F. Santos, pela valiosa ajuda na coleta e montagem do material, pelos momentos e pela amizade. A todos os colegas do Laboratório de Herpetologia do LECZ (Instituto Butantan), em especial a Flora, Betinha, Priscila Pisanelli, Priscila Oliveira, Bruno, Valdir, Gileno e Marcelo pela enorme contribuição em conhecimento e pela amizade e parceria. A Francisco Franco, pela enorme contribuição de conhecimento herpetológico e pela parceria e ajuda. A Bruno Ferreto pela ajuda tanto com material de coleta, como a oportunidade de coletar e conhecer o sul do estado de São Paulo e pelos bons momentos e parceria. A todos os colegas do VPS (FMVZ-USP), em especial o Prof. Marcelo B. Labruna, Arlei Marcili e Fernanda Nieri, por abrir as portas do laboratório e permitir o desenvolvimento das técnicas moleculares, bem como a ampliação do meu conhecimento. A Markéta Nováková, pela ajuda no aprimoramento da técnica de GT que permitiu a extração de DNA de ácaros Trombidiformes, pela amizade e bons momentos. A Marcos Gomes Lopes, Herbert Soares e Solange Oliveira por serem excelentes amigos e companheiros de casa, que contribuíram em cada passo do desenvolvimento deste projeto. Aos amigos Yuri, Caro, Pedro, Sebas, Gonza, Cathe, Vane, Xime, Blanca, Anabella, Margarita, Paula e Dave por serem parte especial da minha vida. A minha família, minha mãe Luz Mery, Meu Pai Hugo meus irmãos Juan e Miguel, por acreditarem em mim e me apoiarem desde o início desta viagem e pelo amor incondicional. A meu irmão gêmeo Miguel Angel, por ser um companheiro desde o início da minha vida, por estar comigo em todos os momentos e me dar ânimos sempre que precisei. Obrigado Hermanito. As agências de fomentos FAPESP, CNPq e CAPES pelo apoio financeiro em projetos e bolsas que contribuíram para execução deste trabalho. Por fim, agradeço a todos que contribuíram neste trabalho, e aqueles que indiretamente, sabendo ou não, estiveram presentes em momentos da minha vida. “Reptiles and amphibians are sometimes thought of as primitive, dull and dimwitted. In fact, of course, they can be lethally fast, spectacularly beautiful, surprisingly affectionate and very sophisticated” (Sir David Frederick Attenborough, 2008) RESUMO MENDOZA-ROLDAN, J. A. Estudos morfológicos e investigação da presença de bactérias e protozoários em ácaros (Trombidiformes), parasitos de répteis e anfíbios, no estado de São Paulo. [Morphological studies and presence research of bacteria and protozoa in mites (Trombidiformes), parasites of reptiles ans amphibians from the state of São Paulo]. 2015. 198 f. Dissertação (Mestrado em Ciências) – Faculdade de Medicina Veterinária e Zootecnia, Universidade de São Paulo, São Paulo. 2015 O Brasil é um país megadiverso em herpetofauna, e do total da diversidade brasileira, aproximadamente 30% das espécies de répteis e 27% das espécies de anfíbios ocorrem no estado de São Paulo. A grande urbanização e o desmatamento acentuado têm ocasionado o aumento de encontros entre a herpetofauna e a população da capital e das outras cidades do estado. Esse fato faz com que algumas espécies, antes florestais, sejam atualmente consideradas sinántropicas. Répteis e anfíbios são amplificadores e reservatórios conhecidos de vários patógenos, mas o papel destes animais no ciclo de doenças e o potencial vetorial dos ácaros ectoparasitas desses vertebrados são ainda pouco conhecidos. Répteis e anfíbios são parasitados por ácaros da ordem Trombidiformes, pertencentes a 5 famílias (Cloacaridae, Pterygosomatidae, Harpirhynchidae, Trombiculidae e Leeuwenhoekiidae). Com exeção de Cloacaridae, as outras ocorrem na América do Sul. Dessa forma, a situação fragmentária dos registros de Trombidiformes no estado de São Paulo, sua complexidade taxonômica e a escassês de informações sobre sua participação na epidemiologia de doenças, foram os principais motivos que levaram à proposição do presente estudo. Os ácaros Trombidiformes de répteis e anfíbios, que estão depositados na coleção acarológica de Instituto Butantan (IBSP) foram revisados e alguns identificados. Igualmente, aqueles ácaros obtidos na recepção de animais do Instituto Butantan e de coletas em campo foram também identificados. Parte do material foi preparada para estudos moleculares e inferência filogenética, usando genes ribossomais e mitrocondriais, e parte foi investigada para a presença de Rickettsia spp., Coxiella spp. e Hepatozoon spp. As seguintes espécies de ácaros foram identificadas, Geckobia hemidactyli, Ophioptes parkeri, O. brevipilis, O. longipilis, Eutrombicula alfreddugesi, Fonsecia ewingi, Hannemania hepatica, H. minor e H. yungicola. Foi descrita uma espécie nova de Ophioptes, sendo a primeira deste gênero parasitando viperídeos. Todos os registros de hospedeiro são novos, assim como, algumas localidades são novos registros de ocorrência para as espécies G. hemidactyli, H. minor e H. yungicola. Foi proposta uma filogenia utilizando-se o gene 18S V4 rRNA da ordem Trombidiformes. As sequências obtidas foram analisadas juntamente com aquelas depositadas no GenBank. Não foi detectada a presença de patógenos nos ácaros e nem nos hospedeiros investigados. Palavras-chave: Taxonomia. Trombidiformes. Répteis. Anfíbios. Filogenia. Patógenos. ABSTRACT MENDOZA-ROLDAN, J. A. Morphological studies and presence research of bacteria and protozoa in mites (Trombidiformes), parasites of reptiles and amphibians from the state of São Paulo. [Estudos morfológicos e investigação da presença de bactérias e protozoários em ácaros (Trombidiformes), parasitos de reptéis e anfíbios, no estado de São Paulo]. 2015. 198 p. Dissertação (Mestrado em Ciências) – Faculdade de Medicina Veterinária e Zootecnia, Universidade de São Paulo, São Paulo. 2015 Brazil is a megadiverse country in herpetofauna, and appoximately 30% of the reptile species, and 27% of the amphibian species ocurr in the state of São Paulo. The high urbanization and the marked deforestation have increase the amount of human-herpetofauna encounters in the capital city and in the enviroment. This fact made some species to be considered currently as sinantropic. Reptiles and amphibians are known amplifiers and reservoirs of several pathogens, yet the role of these animals in the cycle of diseases and the vector potencial of the ectoparasitic mites of these vertebrates are poorly known. Reptiles and amphibians are parasitized by mites of the Trombidiformes order of 5 families (Cloacaridae, Pterygosomatidae, Harpirhynchidae, Trombiculidae e Leeuwenhoekiidae). These families ocurre in South America, except for Cloacaridae. Thereby, the fragmentary records of Trombidiformes in the state of São Paulo, their taxonomic complexity and the scarce information regarding their role in the epidemiology of diseases, were the main reasons
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