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Cnidaria: Myxo UFRRJ INSTITUTO DE VETERINÁRIA CURSO DE PÓS-GRADUAÇÃO EM CIÊNCIAS VETERINÁRIAS TESE Myxozoa Grassé, 1970 (Cnidaria: Myxosporea): Sinopse das espécies parasitando peixes nas Américas e diagnóstico morfológico e molecular das espécies parasitando Characiformes, Leporinus friderici (Anostomidae) e Astyanax altiparanae (Characidae) oriundos do Rio Mogi Guaçú, São Paulo, Brasil Letícia Gabriela Poblete Vidal 2017 UNIVERSIDADE FEDERAL RURAL DO RIO DE JANEIRO INSTITUTO DE VETERINÁRIA CURSO DE PÓS-GRADUAÇÃO EM CIÊNCIAS VETERINÁRIAS MYXOZOA GRASSÉ, 1970 (CNIDARIA: MYXOSPOREA): SINOPSE DAS ESPÉCIES PARASITANDO PEIXES NAS AMÉRICAS E DIAGNÓSTICO MORFOLÓGICO E MOLECULAR DAS ESPÉCIES PARASITANDO CHARACIFORMES, Leporinus friderici (ANOSTOMIDAE) E Astyanax altiparanae (CHARACIDAE) ORIUNDOS DO RIO MOGI GUAÇÚ, SÃO PAULO, BRASIL Letícia Gabriela Poblete Vidal Sob a Orientação do Professor José Luis Fernando Luque Alejos Co-orientação da Professora Rita de Cássia Alves Alcantara de Menezes Tese submetida como requisito parcial para obtenção do grau de Doutora em Ciências, no Curso de Pós Graduação em Ciências Veterinárias.. Seropédica, RJ Março de 2017 UNIVERSIDADE FEDERAL RURAL DO RIO DE JANEIRO INSTITUTO DE VETERINÁRIA CURSO DE PÓS-GRADUAÇÃO EM CIÊNCIAS VETERINÁRIAS Letícia Gabriela Poblete Vidal Tese submetida como requisito parcial para obtenção do grau de Doutora em Ciências, no Curso de Pós-Graduação em Ciências Veterinárias. DEDICATÓRIA Dedico este trabalho aos meus pais Eduardo e Imaculada por me darem a oportunidade de poder estudar e acreditarem no que eu faço. Aos meus irmãos Andrés e Dudu por sempre estarem me apoiarem apesar da distância e ao meu noivo Luiz Henrique pela paciência durante todo esse período de dedicação aos meus estudos. Ter um lugar para ir é lar. Ter alguém para amar é família. Ter os dois é benção.” Autor desconhecido AGRADECIMENTOS Ao meu orientador Prof. Dr. José Luis Fernando Luque Alejos, pela oportunidade de trabalho, pelos ensinamentos e conselhos que além de tornar esse projeto possível, pude crescer profissionalmente e pessoalmente. Aos colegas do Laboratório de Parasitologia de Peixes da UFRRJ, Carla Juliete, Fabiano Paschoal, Felipe Bisaggio, Íris Soares, Juliana Moreira, Maria Catarina, Naraiana Taborda, Philippe Vieira e Sara Coelho pela companhia diária, auxílio de modo totalmente prestativo e amizade. Em especial à minha grande amiga Camila Pantoja pelo seu companheirismo e cumplicidade durante todos os meus anos de dedicação à pós-graduação. Uma amizade que se iniciou durante meu mestrado e que estará sempre comigo. Ao Dr. Christopher Whipps, da State University of New York – ESF, Syracuse, por me receber em seu laboratório durante meu doutorado sanduíche nos EUA. Aos profissionais do Centro Nacional de Pesquisa e Conservação de Peixes Continentais CEPTA/ICMBIO e ao amigo M.Sc. Júlio Cesar C. de Aguiar, pela colaboração nas coletas dos materiais biológicos. A minha família que sempre me apoiou e incentivou a estudar, e que cuida de mim com paciência, amor e confiança. Ao meu noivo Luiz por sempre me apoiar, incentivar, cuidar e estar ao meu lado com amor. A todos os professores e funcionários do Curso de Pós-Graduação em Ciências Veterinárias, UFRRJ. Ao Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq) pela concessão da bolsa de estudo. RESUMO VIDAL, LETÍCIA GABRIELA POBLETE. Myxozoa Grassé, 1970 (Cnidaria: Myxosporea): Sinopse das espécies parasitando peixes nas Américas e diagnóstico morfológico e molecular das espécies parasitando Characiformes, Leporinus friderici (Anostomidae) e Astyanax altiparanae (Characidae) oriundos do rio Mogi Guaçú, São Paulo, Brasil. 2017. 191p. Tese (Doutorado em Ciências Veterinárias). Instituto de Veterinária, Universidade Federal Rural do Rio de Janeiro, Seropédica, RJ. 2017. O presente trabalho teve como objetivo ampliar os conhecimentos sobre a biodiversidade de mixosporídeos parasitos de peixes, visto o escasso conhecimento para esse grupo. No capítulo 1 o objetivo foi fornecer uma sinopse de espécies de Myxozoa Grassé, 1970 nas Américas com base em um levantamento bibliográfico desde 1893, quando a primeira espécie de myxosporídeo foi descrita, até dezembro de 2015. A sinopse inclui uma lista parasito-hospedeiro com dados sobre o habitat do hospedeiro, sítio de infecção, localidade, tamanho e formato do cisto, medidas dos esporos e espécimes em coleções e uma lista de parasitos-hospedeiros. Nessa sinopse são relatados somente as descrições originais encontradas nas Américas. No capítulo 2, é incluída uma descrição de H. friderici Casal, Matos e Azevedo, 2003 com novos dados morfológicos e moleculares com material proveniente de amostras de filamentos branquiais de Leporinus friderici (Bloch, 1794) do rio Mogi Guaçú, estado de São Paulo. Finalmente, no capítulo 3, uma espécie de Henneguya encontrada nos rins de Astyanax altiparanae (Characiformes: Characidae) foi descrita e ilustrada com base na sua morfologia e na análise do rDNA da subunidade maior do ribossomo (28S). Estes dados identificam uma possível espécie nova de Myxozoa. Palavras-chave: Sinopse, Morfologia, Mixosporídeos, 18S (SSU), 28S (LSU) ABSTRACT VIDAL, LETÍCIA GABRIELA POBLETE. Myxozoa Grassé, 1970 (Cnidaria: Myxosporea): Synopsis of species parasiating fish in the Americas and morphological and molecular diagnosis of species parasiating Characiformes species, Leporinus friderici (Anostomidae) and Astyanax altiparanae (Characidae) from the Mogi Guaçú River, São Paulo, Brazil. 2017. 191p. Thesis (PhD in Veterinary Sciense). Instituto de Veterinária, Universidade Federal Rural do Rio de Janeiro, Seropédica, RJ. 2017. The hope of this study was increased the knowledge of biodiversity of myxozoan fish parasites, unknown to some groups. In chapter 1, the goal was provide a synopsis of Myxozoa Grassé, 1970 species in the Americas based on records of the valid species of myxozoans (Myxozoa: Myxosporea) described in the Americas is provided based on a comprehensive survey of the literature since 1893, when the first myxozoan species was reported, until December 2015. The synopsis include the habitat of the host, site of the infection of the parasite, locality, size (µm or mm) and form of the plasmodia, spore measurements, provide specimens to parasitological collections, molecular data and explicit linkage of host. This synopsis was based on original descriptions. In chapter 2, the present work complements the original description of H. friderici Casal Matos and Azevedo, 2003 with new morphological and molecular data with Gill filaments on Leporinus friderici (Bloch, 1794) from the Mogi Guaçú River, state of São Paulo. Finaly, in chapter 3, specie of Henneguya was found in the kidneys of Astyanax altiparanae (Characiformes: Characidae) and were analyzed by morphological and molecular studies with analysis of the rDNA of the small subunid of the ribosome (18S). These data identify a new species of Myxozoa. Key words: Synopsis, Taxonomy, Myxozoans, 18S (SSU), 28S (LSU) LISTA DE TABELA CAPÍTULO I Tabela 1. Espécies relatadas nas Américas que foram originalmente descritas de outros continentes.........................................................................................................................8 Tabela 2. Espécies de mixosporídeos e seus hospedeiros ....................................... .......10 CAPÍTULO II Tabela 1. Lista de mixosporídeos cujas sequências foram utilizadas para análises moleculares e as obtidas no presente estudo (em negrito)…………………………….137 Tabela 2. Comparação das características de Henneguya friderici com espécies semelhantes. CE: comprimento do esporo; LE: largura do esporo; CC: comprimento da cauda; CL: comprimento total do esporo; CCP: comprimento da cápsula polar; LCP: largura da cápsula polar..............................................................................................142 CAPÍTULO III Tabela 1. Lista de mixosporídeos cujas sequências foram utilizadas para análises moleculares e as obtidas no presente estudo (em negrito)…………………………….151 Tabela 2. Comparação das características de Henneguya sp. nov. 1 com espécies semelhantes. CE: comprimento do esporo; LE: largura do esporo; CC: comprimento da cauda; CL: comprimento total do esporo; CCP: comprimento da cápsula polar; LCP: largura da cápsula polar……………………………………………………….…....…155 Tabela 3. Similaridade genética entre espécies de Henneguya depositadas GenBank e as apresentadas no presente estudo. (*) Sequência obtida no presente estudo..................157 LISTA DE FIGURAS CAPÍTULO II Figura 1 Leporinus friderici (Bloch, 1794)..................................................................135 Figura 2 Plasmódio de Henneguya friderici infectando o filamento branquial de Leporinus friderici (Bloch, 1794). Escala de 10 mm....................................................140 Figura 3 Henneguya friderici; a - Esporo maduro de H. friderici parasito de filamentos branquiais de Leporinus friderici em vista frontal em DIC; b – Desenho esquemático do esporo de H. friderici demonstrando a cápsula polar e os prolongamentos caudais bífidos. Escala de 10 μm.. .............................................................................................. 141 Figura 4 Máxima verossimilhança da análise filogenética das sequências do gene 18S rDNA de Henneguya friderici associado com a espécie mais próxima indicada pela análise de Max Score por BLAST da plataforma NCBI. O primeiro número de suporte nodal é de bootstrap de máxima verosimilhança (1000 repetições), o segundo número mostra a análise
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