Orientador: João Manuel Grisi Candeias / Resumo: Não disponível. / Abstract: From about 200 virus that cause respiratory infections, only 8 are responsible for severe illness in children, immunodeficient adults and elderly, inc1uding Adenovirus, Influenza A and B, Parainfluenza 1, 2 and 3, Human RespÍratory Syncytial Vírus (HRSV) and Human Metapneumovirus (hMPV). Three of these virus are responsible for a significant morbidity in elderly: Influenza, HRSV and hMPV. The nosocomial infection caused by these vÍruses can be fatal in hospitalized children and patients with other pathologies. With the advance of the of molecular biology techniques, the diagnosis and the characterization of these vírus became more effective. Beside vírus detection by PCR, isolation of vírus in specific cellular cultures to increase the amount of pathogens have been used in severallaboratories. Studies on prevalence of respÍratory vírus in elderly are rare. The objective of this research was to evaluate the occurrence of Influenza, HRSV and of the hMPV and the risk factors involved in the diseases caused by these vÍruses. Nasopharyngeal swabs were collected and used for inoculation in cells culture and dÍrect analysis by RT-PCR. The results from RT-PCR of Influenza Vírus and RSV were negative. We also tested the samples with GeneScan, that is a technique based on fluorescent primers specific to the studied vírus. Results for Flu virus and HRSV were the same ofRT-PCR, but 1 MPV sample was positive. 55,32% ofthe 47 studied elderly were vaccinated against Influenza; 14,9% had tabagism habits and 70,2% were women. These risk factors might had influenced in the absence of positive samples for the Influenza vírus andHRSV. / Mestre

Orientador: Antonio Carlos Paes / Banca: Marcio Garcia Ribeiro / Banca: Hélio Langoni / Banca: Christian Hirsch / Banca: Osimar de Carvalho Sanches / Resumo: O estudo objetivou identificar efeitos do tratamento com ribavirina, prednisona e DMSO na cinomose; identificar a presença viral no sangue, medula óssea e líquor antes e após o tratamento, os efeitos colaterais dos fármacos experimentais e associações. Foram utilizados 60 cães apresentando sinais neurológicos da cinomose com evolução de dez dias. Animais foram internados para tratamento de suporte; avaliados diariamente e submetidos ao hemograma e dosagens bioquímicas. Os grupos 1 e 2 foram tratados com ribavirina associada ao DMSO; os grupos 3 e 4 foram tratados com DMSO e prednisona e os grupos 5 e 6 foram tratados com ribavirina e prednisona, ribavirina, prednisona e DMSO. Os animais foram anestesiados e colhidos líquor, medula óssea e sangue, antes e após o tratamento e realizada a RTPCR das amostras; as negativas foram analisadas pela técnica de hn-PCR. O vírus foi encontrado em 95% das amostras de sangue, 90% de medula óssea e 53,3% de líquor pré-tratamento. O efeito adverso da ribavirina quando associada com a prednisona foi anemia. A prednisona na dose imunossupressora causou aumento da dosagem de proteína e diminuição da celularidade liquórica, leucocitose. Já a dose antinflamatória causou diminuição de proteína no líquor. Baseado nos índices de sobrevida e melhora clínica, o tratamento mais efetivo foi o G2 (80%); seguido do G1, G5 e G3 (70%); o G6 (60%); o G4 com o pior índice (30%). Pós-tratamento, a frequência viral foi 97,7% no sangue, 86,4% na medula óssea e 27,3% no líquor / Abstract: The present study aims at the identification of ribavirin, prednisone and DMSO's treatment effects in dogs with canine distemper, at the identification of the viral presence in the blood, bone marrow and cerebrospinal fluid (CSF) before and after the treatment and also at the identification of side effects of the experimental drugs and its combinations. Sixty dogs presenting canine distemper with neurological signs about ten days evolution were observed. The animals were hospitalized for the support treatment, assessed on daily basis and subjected to blood cells count and biochemical analysis. Groups 1 and 2 were treated with ribavirin and its combination with DMSO; Groups 3 and 4 treated with prednisone and DMSO, Group 5 treated with ribavirin and prednisone, while Group 6 with ribavirin, prednisone and DMSO. The animals were anesthetized for the cerebrospinal fluid, bone marrow and blood samples collection before and after the treatment, then the RT-PCR of the samples was proceeded. The negative were analysed according to the hn-PCR technique. The canine distemper virus were found in 95% of blood samples, 90% of bone marrow and 53,3% of CSF before the treatment. The adverse effect of ribavirin and its association with prednisone was anemia. Prednisone, at its immunosuppressive dose, led to the increase of protein and decrease of cellularity in CSF, and increase of leukocytes blood count. The antiinflammatory dose led to the CSF protein concentration's decrease. Considering the survival and clinical improvement rates, the most successful treatment was the one applied to the G2 (80%); followed by G1 (70%); G5 (70%) and G3 (70%); G6 (60%); and the lowest rate G4 (30%). After the treatment, the virus frequency was 97,7% in the blood, 86,4% in the bone marrow and 27,3% in the CSF / Doutor

Orientador: Hélio José Montassier / Banca: Clarice Weins Arns / Banca: Adolorata Aparecida Bianco Carvalho / Resumo: A bronquite infecciosa das galinhas (BIG) é uma doença infecciosa que está amplamente disseminada entre as criações avícolas brasileiras e é uma das enfermidades virais que mais têm causado perdas econômicas na atualidade. Portanto, a rápida detecção e identificação do agente causal são imprescindíveis para que medidas eficazes de controle sejam prontamente tomadas. Para tanto, é necessário que os métodos de diagnóstico empregados sejam sensíveis, específicos, rápidos e também de baixo custo. Nesse contexto, a técnica de RTPCR em tempo real abordada no presente estudo permitiu a amplificação de duas regiões de hipervariabilidade do gene S1 de 17 estirpes diferentes do vírus da BIG (VBI), que foram testadas, mas não foi capaz de amplificar nenhum dos RNAvírus heterólogos analisados (vírus da doença de Newcastle, pneumovírus aviário e vírus da doença de Gumboro). Com essa mesma técnica foi possível fazer a diferenciação em grupos geneticamente distintos, de estirpes do VBI através de análises das curvas de dissociação de fragmentos amplificados a partir das regiões de hipervariabilidade gênica I e II do gene S1. A RT-PCR em tempo real desenvolvida apresentou maior sensibilidade na detecção do VBI em amostras teciduais, quando comparada à técnica padrão de Isolamento Viral em ovos embrionados de galinha... (Resumo completo, clicar acesso eletrônico abaixo) / Abstract: The avian infectious bronchitis virus (IBV) is an infectious disease widely spread in Brazilian commercial poultries where causes significant economical losses. Rapid and accurate diagnosis of the IBV strain involved in a field outbreak is necessary to establish an effective control of this disease. The real-time RT-PCR performed in this study to amplify two hypervariable regions of S1 gene, was able to detect 17 IBV strains, e.g., nine reference strains (including Massachussets, Connecticut, JMK, SE 17 and Iowa serotypes) and eight Brazilian field isolates, whilst non-related avian viral pathogens such as Newcastle disease virus, Avian Pneumovirus and Gumboro disease virus were not detected. The differentiation between IBV strains was accomplished using the melting curve analysis of the amplified fragments corresponding to the hypervariable regions I and II of S1 gene. The real-time RT-PCR developed here showed a higher rate of IBV detection in tissue samples of experimentally infected chickens, when compared to the goldstandard technique of Viral Isolation in embryonated chicken eggs, and the same rate of detection was found for the conventional RT-PCR... (Complete abstract, click electronic access below) / Mestre