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Doctoral Thesis
DOI
10.11606/T.10.2005.tde-18102006-123602
Document
Author
Full name
Enio Mori
E-mail
Institute/School/College
Knowledge Area
Date of Defense
Published
São Paulo, 2005
Supervisor
Committee
Fernandes, Wilson Roberto (President)
Benesi, Fernando José
Gonçalves, Roberto Calderon
Lara, Maria do Carmo Custodio de Souza Hunold
Richtzenhain, Leonardo Jose
Title in Portuguese
Infecção experimental em cavalos pelo herpesvírus eqüino tipo 1: aspectos clínicos e detecção do agente pela reação em cadeia pela polimerase
Keywords in Portuguese
DNA viral
Eqüinos
Herpesviridae
Reação em cadeia por polimerase
Rinopneumonite animal
Abstract in Portuguese
Dez cavalos adultos clinicamente saudáveis foram inoculados por via intranasal com a estirpe A4/72 do herpesvírus eqüino tipo 1 (HVE-1). Com o intuito de estudar o efeito da dose infectante na severidade da rinopneumonite, os animais foram distribuídos em dois grupos experimentais: (a) grupo I (106,6 DICT50) e (b) grupo II (5×106,6 DICT50). Nos primeiros dez dias após a inoculação viral, todos os cavalos apresentaram manifestações de infecção respiratória leve e restrita às vias aéreas anteriores. Poucos animais desenvolveram leucopenia sangüínea envolvendo linfócitos (n=4) e neutrófilos (n=2). Somente em um houve aumento na contagem dos neutrófilos no lavado broncoalveolar (LBA). Apesar de possuírem elevados títulos de anticorpos neutralizantes antes da inoculação, alguns cavalos apresentaram soroconversão após o desafio viral. Esse padrão de resposta humoral foi determinado pelo aumento da dose infectante. O HVE-1 não foi isolado a partir das secreções nasais de nenhum animal. Entretanto, o DNA viral foi detectado pela reação em cadeia pela polimerase (PCR) nas células mononucleares sangüíneas entre o terceiro e o oitavo dias pós-inoculação (d.p.i.) em todos os animais, indicando a ocorrência de viremia. Além disso, a prova de PCR detectou o vírus nas amostras de LBA a partir do nono d.p.i. no grupo II, demonstrando que a disseminação do HVE-1 pelo trato respiratório após o desafio viral foi dose-dependente. Com base nos resultados obtidos, foi possível concluir que a PCR é uma técnica com alta sensibilidade para o diagnóstico do HVE-1, capaz de detectar a presença do DNA viral mesmo quando não ocorre a constatação do agente pelos métodos tradicionais
Title in English
Experimental infection in horses with equine herpesvirus 1: evaluation of clinical signs and detection of virus by polymerase chain reaction
Keywords in English
Animal rhinopneumonitis
Equine
Herpesviridae
Polymerase chain reaction
Viral DNA
Abstract in English
Ten clinically healthy adult horses were inoculated intranasally with the A4/72 strain of equine herpesvirus 1 (EHV-1). The animals were divided into two experimental groups in order to study the influence of the infective dose in the severity of rhinopneumonitis: (a) group I (106,6 TCID50) and (b) group II (5×106,6 DICT50). In the first ten days after the inoculation, they showed signs of a mild, self-limiting upper respiratory tract infection. Very few animals developed transient blood leukopenia, involving lymphocytes (n=4) as well as neutrophils (n=2). Only one horse had an increase in the neutrophils count of the bronchoalveolar lavage (BAL) samples. In spite of the presence of neutralizing antibodies before the trial, seroconversion was observed in some horses. The pattern of antibody response was determined by the increase of the challenge exposure. The virus was not isolated from nasal swabs of any horse. However, the EHV-1 was detected through the polymerase chain reaction (PCR) from peripheral blood mononuclear cells (PBMC) of all horses in the experiment within the third to the eighth day after the inoculation that illustrated the viremia. In addition, the PCR assay also detected the virus in BAL samples starting on the ninth day after the experimental infection of group II. For that reason, the dissemination of the EHV-1 throughout the respiratory tract after virus exposure was dose-dependent. Based on the results obtained from this study, it can be affirmed that the PCR is a highly effective technique in detecting the EHV-1. It may be used in circumstances where traditional methods are not efficient due to the fact that it provides an enhanced diagnostic procedure for underdiagnosed diseases
 
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Publishing Date
2006-11-17
 
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