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I Secretaria De Agricultura E Abastecimento Do Estado De São Paulo Agência Paulista De Tecnologia Dos Agronegócios Instituto i Secretaria de Agricultura e Abastecimento do Estado de São Paulo Agência Paulista de Tecnologia dos Agronegócios Instituto Biológico Programa de Pós-Graduação em Sanidade, Segurança Alimentar e Ambiental no Agronegócio Prevalência e caracterização espacial de sorotipos do vírus da língua azul em bovinos, São Paulo, Brasil, 2011 Thaís Garcia da Silva Tese apresentada para a obtenção do título de Doutora em Sanidade, Segurança Alimentar e Ambiental no Agronegócio. Área de concentração: Segurança Alimentar e Sanidade no Agroecossistema. São Paulo 2017 ii Thaís Garcia da Silva Prevalência e caracterização espacial de sorotipos do vírus da língua azul em bovinos, São Paulo, Brasil, 2011 Tese apresentada para a obtenção do título de Doutora em Sanidade, Segurança Alimentar e Ambiental no Agronegócio. Área de concentração: Segurança Alimentar e Sanidade no Agroecossistema. Orientadora: Profa. Dra. Edviges Maristela Pituco iii Eu, Thaís Garcia da Silva, autorizo o Instituto Biológico (IB-APTA), da Secretaria de Agricultura e Abastecimento do Estado de São Paulo, a disponibilizar gratuitamente e sem ressarcimento dos direitos autorias, o presente trabalho acadêmico, de minha autoria, no portal, biblioteca digital, catálogo eletrônico ou qualquer outra plataforma eletrônica do IB para fins de leitura, estudo, pesquisa e/ou impressão pela Internet desde que citada a fonte. Assinatura: _______________________________ Data 05/ 07/ 2017 Dados Internacionais de Catalogação na Publicação (CIP) Secretaria de Agricultura e Abastecimento do Estado de São Paulo Núcleo de Informação e Documentação – IB Silva, Thaís Garcia da. Prevalência e caracterização espacial de sorotipos do vírus da língua em bovinos, São Paulo, Brasil, 2011. / Thais Garcia da Silva. - São Paulo, 2017. 121 p. Tese (Doutorado). Instituto Biológico (São Paulo). Programa de Pós- Graduação. Área de concentração: Segurança Alimentar e Sanidade no Agroecossistema. Linha de pesquisa: Gestão Sanitária e Ambiental na produção animal. Orientador: Edviges Maristela Pituco. Versão do título para o inglês: Prevalence and spatial characterization of blue - tonge in cattle from São Paulo State, Brazil, 2011. 1. Orbivirus 2. Reoviridae 3. Sorologia 4. Virusneutralização I. Silva, Thais Garcia da. II. Pituco, Edviges Maristela. III. Instituto Biológico (São Paulo). IV. Título. IB/Bibl./2017/012 iv FOLHA DE APROVAÇÃO Nome: Thaís Garcia da Silva Título: Prevalência e caracterização espacial de sorotipos do vírus da língua azul em bovinos, São Paulo, Brasil, 2011. Tese apresentada ao Programa de Pós-graduação em Sanidade, Segurança Alimentar e Ambiental no Agronegócio do Instituto Biológico, Agência Paulista de Tecnologia dos Agronegócios, da Secretaria de Agricultura e Abastecimento do Estado de São Paulo para a obtenção do título de Doutora em Sanidade, Segurança Alimentar e Ambiental no Agronegócio. Aprovado em: 05/07/2017 Banca Examinadora: Profa. Dra. Edviges Maristela Pituco Instituição: Instituto Biológico Julgamento:___________________________ Assinatura:_________________ Profa. Dra. Claudia Del Fava Instituição: Instituto Biológico Julgamento:____________________________ Assinatura:_____________________ Profa. Dra. Dolores Ursula Mehnert Instituição: Universidade de São Paulo Julgamento:_____________________________ Assinatura:_____________________ Profa. Dra. Alice M.M.P. Della Libera Instituição: Universidade de São Paulo Julgamento:_____________________________ Assinatura:_____________________ Prof. Dr. Leonardo José Richtzenhain Instituição: Universidade de São Paulo Julgamento:_____________________________ Assinatura:______________________ v DEDICATÓRIA Meu Quilombo, minha fortaleza: Jorge, Laudimara, Fernanda, Débora, Alamim e Heitor vi AGRADECIMENTOS À Profª Drª. Edviges Maristela Pituco, por me receber em seu Laboratório, e também na Pós- Graduação do Instituto Biológico, pela orientação, confiança e por oferecer todas as condições necessárias para o desenvolvimento desta pesquisa. Ao Dr. Giovanni Savini pela orientação durante o período de doutorado sanduíche no Istituto Zooprofilattico Sperimentale dell'Abruzzo e del Molise "G. Caporale" (IZSAM), Teramo, Itália, e por disponibilizar a estrutura laboratorial e banco de padrões virais de sorotipos do BTV, viabilizando a execução desta pesquisa. À querida Profª. Drª. Claudia Del Fava, pela amizade, conselhos, ajuda inestimável e, principalmente, pelos ensinamentos, que levarei para minha vida profissional. Aos médicos veterinários da Coordenadoria de Defesa Agropecuária pela colheita das amostras e por disponibilizá-las ao Laboratório de Viroses de Bovídeos para esta e outras pesquisas. À M.Sc. Michele dos Santos Lima, pelo profissionalismo, amizade, companheirismo, desde a concepção do projeto até as análises dos resultados. Pelo estímulo e motivação nos momentos bons e ruins. Às amigas, Maira Souza, Vívian Cardoso, Larissa de Paula e Adeline Fernandes, pela convivência, encontros, ajuda nos momentos de dificuldades e companheirismo. À equipe do Departamento de Virologia do IZSAM: Massimo Spedicato, Irene Carmine, Liana Teodori, Alessandra Leoni, Annapia Di Gennaro, Maura Picella, Claudia Casaccia, Miriam Berti Sabrina Olivieri, Barbara Cipro, Daniela Malatesta, Guendalina Zaccaria, Valeria Marini, Ottavio Portanti e Alessio Lorusso, pelo aprendizado, ótimo convívio, disponibilidade e colaboração. Aos amigos Vincenzo D’Innocenzo, Mirella Luciani, Micheli Podaliri, Dani, Luigi Iannett, Stafania Grilli, que tornaram um ano longe de casa muito mais especial: “bella situazione”. Às funcionárias, pesquisadoras e estudantes do Laboratório de Viroses de Bovídeos, Cláudia Pestana, Taís Caiuby, Marta Vicente, Alisete Assumpção, Líria Okuda, Adriana Nogueira, Eliana De Stefano, Antera Ferreira, Regina Paixão, Bianca Freire, Vinícius Scenegaglia e Priscilla Felício, pela colaboração. Ao M.Sc. Flávio Henrique Mendes, pela colaboração nas análises climáticas, ao Dr. Alexandre L. R. Chaves, pela elaboração da Figura 1 desta Tese, e ao Roberto Tadeu pelo auxílio na biblioteca. Ao Programa de Pós-Graduação do Instituto Biológico, ao Dr. Marcelo Eiras, Samira Farah Pizolato, aos docentes, funcionários, e também aos colegas de curso. Aos membros da banca examinadora, Profª. Drª. Alice M.M.P. Della Libera, Profª Drª. Claudia Del Fava, Profª Drª. Dolores Ursula Mehnert e Prof. Dr. Leonardo José Richtzenhain, pelas correções, críticas e sugestões que enriqueceram este trabalho. À CAPES (Coordenação de Aperfeiçoamento de Pessoal de Nível Superior) pela bolsa de Doutorado e pela bolsa do PDSE (Programa de Doutorado Sanduíche no Exterior), apoio financeiro fundamental para a realização dos trabalhos no Brasil e no exterior. vii EPÍGRAFE “...Por que não queria ser margem Queria mesmo ser mar...” Débora Garcia “A concordância é preguiçosa e a discordância é provocadora.” Eduardo Portella viii RESUMO SILVA, Thaís Garcia. Prevalência e caracterização espacial de sorotipos do vírus da língua azul em bovinos, São Paulo, Brasil, 2011. 2017. 121 f. Tese (Doutorado em Sanidade, Segurança Alimentar e Ambiental no Agronegócio) – Instituto Biológico, Agência Paulista de Tecnologia dos Agronegócios, Secretaria de Agricultura e Abastecimento do Estado de São Paulo, São Paulo, 2017. O Bluetongue virus (BTV), gênero Orbivirus, família Reoviridae, apresenta partículas icosaédricas com diâmetro entre 60 e 85 nm. O genoma é constituído por dez segmentos de RNA de fita dupla, e apresenta elevada variabilidade genética, que, em parte, se traduz nos 29 sorotipos de BTV já identificados em diversos países. Mosquitos do gênero Culicoides respondem pela transmissão do vírus, que induz a doença conhecida como língua azul (LA), que pode afetar espécies de ruminantes. A LA é de notificação obrigatória e restringe a movimentação e comércio de animais em áreas livres. No Brasil, os bovinos infectados, normalmente, são assintomáticos com longos períodos de viremia (meses). A associação de temperatura e umidade favoráveis ao desenvolvimento do vetor, e bovinos como fonte de infecção, garante a manutenção do vírus nos rebanhos. Este trabalho teve como objetivos: (i) identificar anticorpos neutralizantes para os sorotipos do BTV; (ii) realizar levantamento da distribuição dos sorotipos de BTV no Estado de São Paulo; (iii) realizar estudo de análise de risco para LA com base em práticas zootécnicas e sanitárias; e (iv) estudar a influência de fatores climáticos e associá-los aos sorotipos identificados. O Estado de São Paulo foi dividido em sete circuitos produtores de bovinos, tendo sido selecionado um animal reagente ao ensaio de ELISA por propriedade (de um total de 1.598 propriedades) e os soros foram submetidos à técnica de virusneutralização contra 25 sorotipos de BTV. Os fatores de risco foram analisados a partir de questionários com variáveis zootécnicas e sanitárias de cada propriedade, empregando análise univariada e regressão logística. A variável aquisição de bovídeos foi considerada fator de risco para a ocorrência da doença (OR = 2,183; IC 95% = 1,6-2,9). Nos circuitos 1, 2 e 3 foram observadas as maiores prevalências de BTV, sendo que no circuito 1, registraram-se as temperaturas mais elevadas do Estado. No presente trabalho, foram identificadas soroprevalências para 11 sorotipos: BTV-1 (22,15%), BTV-2 (31,03%), BTV-3 (18,96%), BTV-4 (24,90%), BTV-9 (6,82%), BTV-12 (7,50%), BTV-17 (23,90%), BTV-19 (10,20%), BTV-21 (30,66%), BTV-22 (12,14%), BTV-26 (57,00%). Neste trabalho, foi identificada pela primeira vez no Brasil evidência sorológica para os sorotipos BTV-1,
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