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Doctoral Thesis
DOI
https://doi.org/10.11606/T.10.2003.tde-10052004-140909
Document
Author
Full name
Marcia Ester Parreira Vasconcellos
Institute/School/College
Knowledge Area
Date of Defense
Published
São Paulo, 2003
Supervisor
Committee
Ito, Fumio Honma (President)
Cortes, Jose de Angelis
Girio, Raul José da Silva
Miguel, Omar
Samara, Samir Issa
Title in Portuguese
Caracterização biológica, genética e sorológica de uma amostra de vírus da raiva isolada de eqüino de uma região próxima de São Paulo, Sudeste do Brasil
Keywords in Portuguese
Equinos
Raiva animal
Vacinas
Abstract in Portuguese
O comportamento biológico de uma amostra de vírus recém-isolada de eqüino (M82-02), procedente de uma região próxima de São Paulo, foi estudado em camundongos, inoculados pelas vias intracerebral e intramuscular, avaliando-se as características relacionadas com a infectividade, patogenicidade, período de incubação, período do curso clínico e capacidade invasiva do vírus para outros tecidos não-nervosos. A amostra, com uma passagem em cérebro de camundongos, foi tipificada antigenicamente, utilizando um conjunto de anticorpos monoclonais (MABs), desenvolvido pela "Canadian Food and Inspection Agency", de Ottawa, Canadá, correspondendo ao perfil de morcego hematófago Desmodus rotundus. Após ser submetida à extração do material genético, a amostra foi caracterizada geneticamente no "National Institute of Infectious Diseases", de Tóquio, Japão, confirmando pertencer ao genótipo 1 do gênero Lyssavirus, com característica próxima a do "Vampire -bat related virus-VRRV", comuns entre as amostras brasileiras isoladas de herbívoros e morcegos hematófagos. As sucessivas passagens da amostra em camundongos, por via intracerebral e intramuscular, provocaram ligeiro aumento no título do vírus e estabilização do período de incubação, quando então foi utilizado como antígeno para a prova de soroneutralização, para avaliar o comportamento deste vírus frente aos soros de eqüinos vacinados com uma vacina comercial de vírus PV inativado. Os soros de eqüinos vacinados foram titulados em paralelo com o antígeno viral constituído de vírus CVS. Com o uso da amostra M82-02 como antígeno para a prova de soroneutralização não foi possível demonstrar diferença significativa, quando comparado com o antígeno fixo de vírus CVS, no entanto, alguns soros de eqüinos vacinados foram encontrados com níveis de anticorpos acima de 0,5UI/mL. Pela prova de imunofluorescência direta (IFD), a presença do antígeno do vírus da raiva foi evidenciada no pulmão e nos rins de animais inoculados por via IM, notadamente nos materiais de passagem mais elevada. No teste de avaliação da vacina comercial, pelo método do CDC de Atlanta, o resultado do desafio com a amostra M82-02 foi inferior ao do vírus fixo CVS, no entanto, este método ainda necessita ser melhorado para sua utilização na rotina.
Title in English
Biologic, genetic and serologic characterization of an equine rabies virus isolated from a neighbor region of São Paulo, Southeastern Brazil
Keywords in English
Animal rabies
Equines
Vaccines
Abstract in English
The biologic behavior of a rabies virus recently isolated from an equine raised at a region neighbor to São Paulo, Southeast Brazil, the isolate M82-02, was studied in mice by inoculating through intracerebral and intramuscular route, for evaluation of characteristics related to its infectivity, pathogenicity, incubation period and course of clinical illness and also the virus capacity to invade other non-nervous tissues. The first intracerebrally passaged mouse brain was submitted for antigenic typing by using a set of monoclonal antibodies (MABs) prepared at the Canadian Food and Inspection Agency, Ottawa, Canada, and it was found to be a variant of vampire-bat related rabies virus. After extraction of genetic material, the sample was characterized genetically at the National Institute of Infectious Diseases, Tokyo, Japan, ant the isolate belonged to genotype 1 of the Lyssavirus gene, closely related to the Vampire-bat related virus group-VRRV, common among the Brazilian rabies virus isolates from herbivores and vampire bat Desmodus rotundus. The successive intracerebral and intramuscular passages in mice provoked a slight increase in the virus titer, and the stabilization of the incubation period. In its 10th passage, the isolate was used as the antigen in mouse serum neutralization test, for the assessment of equine sera which had been vaccinated with a commercial PV rabies inactivated virus vaccine, the equine sera were tested in parallel with the CVS strain and some sera were found with neutralizing titer >0.5 IU/mL. Using the isolate M82-02 of rabies virus as an antigen in the neutralization test, no significant difference could be detected, when the results were compared to that of the CVS strain. By means of direct fluorescent antibody (dFA) test, the presence of rabies virus antigen was detected in tissues of brain, lung and kidneys of mice inoculated by intramuscular route, especially in serially passaged materials. The CDC-potency test for evaluation of a commercial vaccine using the field isolate M82-02 as a challenge virus showed a poorer result than the challenge with the fixed CVS strain, however, this method needs further modifications for the routine use.
 
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Publishing Date
2007-05-02
 
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