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PRESENÇA DE Ureaplasma Diversum EM VACAS DE LEITE PERTENCENTES a MUNICÍPIOS DO NORTE DE MATO GROSSO, BRASIL

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PRESENÇA DE Ureaplasma Diversum EM VACAS DE LEITE PERTENCENTES a MUNICÍPIOS DO NORTE DE MATO GROSSO, BRASIL UNIVERSIDADE FEDERAL DE MATO GROSSO FACULDADE DE AGRONOMIA, MEDICINA VETERINÁRIA E ZOOTECNIA PROGRAMA DE PÓS-GRADUAÇÃO EM CIÊNCIAS VETERINÁRIAS PRESENÇA DE Ureaplasma diversum EM VACAS DE LEITE PERTENCENTES A MUNICÍPIOS DO NORTE DE MATO GROSSO, BRASIL. JAQUELINE BRUNING AZEVEDO Cuiabá – MT 2016 UNIVERSIDADE FEDERAL DE MATO GROSSO FACULDADE DE AGRONOMIA, MEDICINA VETERINÁRIA E ZOOTECNIA PROGRAMA DE PÓS-GRADUAÇÃO EM CIÊNCIAS VETERINÁRIAS PRESENÇA DE Ureaplasma diversum EM VACAS DE LEITE DE DIFERENTES MUNICÍPIOS DO NORTE DE MATO GROSSO, BRASIL. Autor (a): Jaqueline Bruning Azevedo Orientadora: Profª. Drª. Caroline Argenta Pescador Co-Orientadora: Profª. Drª. Valéria Dutra Dissertação apresentada ao Programa de Pós- graduação em Ciências Veterinárias, área de concentração: Sanidade Animal, da Faculdade de Agronomia e Medicina Veterinária da Universidade Federal de Mato Grosso para a obtenção do título de Mestre em Ciências Veterinárias. Cuiabá – MT 2016 AGRADECIMENTOS Neste momento de grande emoção, agradeço aos meus pais, Edemundo e Leoni. Agradeço as noites em claro dirigindo um carro e “dirigindo” uma maquina de costura. Sou o que sou, graças a vocês, meus grandes amores. Agradeço as minhas amadas irmãs Juliana e Janaina, que fazem parte, mesmo que de longe, desta conquista. Vocês todos são a minha essência, o motivo de minha caminhada e a razão de todo esforço. Ao meu amado, Adriano, que me ajudou nos momentos de duvidas e questionamentos. A minha orientadora, profa. Dra. Caroline Argenta Pescador, sempre paciente e a disposição. Foi um período de muito aprendizado e de conhecimento, muito obrigada. A minha co-orientadora, profa. Dra. Valéria Dutra, pelo suporte. A Letícia, que, com muita paciência, ajudou durante todas as PCR’s. Aos amigos que fiz no Laboratório de Patologia Veterinária, desejo tudo de melhor na vida pessoal e profissional de vocês. Em especial, Kassia, Samara e Leilane, que se tornaram grandes amigas. Aos colegas da turma de mestrado e professores. Pelos momentos alegres e de conhecimento. Aos produtores, que abriram as portas de suas propriedades e casas, nos ajudando a tornar possível este trabalho. Meus sinceros agradecimentos. Talvez tenha deixado passar alguns momentos e pessoas, mais o carinho e as lembranças estão guardadas. E por fim e mais importante, agradeço a Deus e Nossa Senhora pelas graças que venho recebendo durante toda vida, esta é mais uma delas. RESUMO PRESENÇA DE Ureaplasma diversum EM VACAS DE LEITE PERTENCENTES A MUNICÍPIOS DO NORTE DE MATO GROSSO, BRASIL A infecção por Ureaplasma diversum em fêmeas bovinas pode resultar em vários problemas reprodutivos, que incluem vulvovaginite granular, bezerros fracos, salpingite e aborto espontâneo. A presença de U. diversum na população bovina leiteira do Centro-Oeste do Brasil não foi estabelecida. O objetivo deste estudo foram determinar se U. diversum está presente em bovinos leiteiros do Centro- Oeste, Brasil, utilizando reação em cadeia da polimerase (PCR). A mucosa vulvovaginal de 203 animais localizados em seis municípios da região Norte de Mato Grosso, Brasil foram analisados. Um total de 25% das vacas leiteiras com vulvovaginite foi positivo pela PCR para U. diversum. Os fatores avaliados foram incluídos em um modelo de regressão logística multivariada, e a variável dependente foi à presença de pelo menos um animal positivo no rebanho. Três variáveis foram significativamente associadas com a PCR positiva para o U. diversum. Foi incluído no modelo multivariada final: número de partos, lesões vulvares e problemas reprodutivos. Cada novo parto, as chances de infecção U. diversum diminuíram 0,03 vezes, indicando que vacas com maior número de partos são mais protegidas. A presença de lesões vulvares foi 17,6 vezes maior para fêmeas positivas para U. diversum, sugerindo que esta bactéria pode estar relacionada à lesão granular vermelha na mucosa vulvar, a chance é 7,6 vezes maiores em vacas com problemas reprodutivos. No entanto, novas investigações devem ser realizadas para verificar os efeitos do U. diversum em associação com outras espécies de Mycoplasmas nos rebanhos estudados. Palavras-chave: fatores associados; leite; PCR vaca; vulvovaginites. ABSTRACT PRESENCE OF Ureaplasma diversum IN MILK COWS BELONGING OF MUNICIPALITIES FROM NORTH OF MATO GROSSO STATE, BRAZIL Ureaplasma diversum infection in bovine females may result in various reproductive problems, including granular vulvovaginitis, abortion, weak calves, salpingitis and spontaneous abortion. The presence of U. diversum in a dairy bovine population from midwestern Brazil has not been established. The aim of this study was to determine whether U. diversum was present in dairy cattle from midwestern Brazil using polymerase chain reaction (PCR). Vulvovaginal mucus was analyzed from 203 animals located in six municipalities in the north region of Mato Grosso State, Brazil. A total of 25% of dairy cows with vulvovaginitis were positive for U. diversum. The factors evaluated were included in a multivariable logistic regression model with the presence of at least one positive animal in the herd serving as the dependent variable. Three variables were significantly associated with a U. diversum positive PCR and were included in the final multivariable model: number of deliveries, vulvar lesions and reproductive problems. For each new delivery, the chance of U. diversum infection decreased 0.03-fold, indicating that cows with the highest number of deliveries were more protected. The presence of vulvar lesions was increased 17.6-fold in females positive for U. diversum, suggesting that this bacteria could be related to the red granulate lesions in the vulvar mucosa, whereas reproductive problems were increased 7.6-fold. However, further investigations should be conducted to ascertain the effects of U. diversum in association with other mycoplasma species in the herds studied. Keywords: associated factors; cow; milk; PCR; vulvovaginitis. LISTA DE ILUSTRAÇÕES Figura 1: Cultura microbiológica do Ureaplasma diversum. A cor da colônia é resultado de uma reação bioquímica entre o sulfato de manganês, que em contato com a amônia, produz o pigmento cor de “ouro” (cortesia: Maristela V. Cardoso – Instituto biológico)..................................................................18 Figura 2 A e B: Rebanhos de bovinos de leite visitados durante o estudo................26 Figura 3 Fêmea bovina. Mucosa vaginal. Vulvovaginite granular grau 1...................26 LISTA DE TABELAS Quadro 1. Principais eventos relacionados ao estudo de U. diversum em bovinos, a partir do ano 2000.....................................................................................15 Tabela 1. Resultado da PCR para U. diversum, associando o número de vacas de leite que apresentaram ou não a lesão de vulvovaginite granular............29 Tabela 2. Presença de U. diversum nos rebanhos bovinos localizados no médio norte do estado de Mato Grosso, Brasil....................................................30 Tabela 3. Número de PCR positivas para U. diversum agrupadas nos 6 municípios localizados no médio norte do Estado de Mato Grosso, Brasil................31 Tabela 4. Resultado da regressão logística das variáveis para os fatores associados ao resultado positivo para U. diversum através da PCR...........................31 ANEXOS Anexo 1: Inquérito epidemiológico aplicado em cada uma das 22 propriedades rurais visitadas.....................................................................................................35 Anexo 2: Comprovante de submissão do artigo na revista Tropical Animal Heath And Production.................................................................................................37 Anexo 3: Artigo submetido a revista Tropical Animal Health And Production..........38 Anexo 4: Certificado de aprovação no Comitê de Ética no uso animal.....................51 SUMÁRIO 1. INTRODUÇÃO ...................................................................................................... 12 2. REVISÃO BIBLIOGRÁFICA .................................................................................. 14 2.1 Histórico ........................................................................................................... 14 2.2. Biologia do agente .......................................................................................... 17 2.3. Patogenia e fatores de risco ........................................................................... 19 2.4. Importância clínica e econômica ..................................................................... 22 2.5. Diagnóstico laboratorial .................................................................................. 24 3. MATERIAL E MÉTODOS ...................................................................................... 25 3.1. População estudada ....................................................................................... 25 3.2. Delineamento amostral e coleta de amostras ................................................. 26 3.3. Inquérito epidemiológico ................................................................................. 28 3.4. Extração de DNA e PCR ................................................................................. 28 4. RESULTADOS ...................................................................................................... 29 5. DISCUSSÃO ......................................................................................................... 32 6. CONCLUSÃO .......................................................................................................
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