Produção e avaliação da proteína derivada (PPD) de Burkholderia mallei para o diagnóstico imuno-alérgico do mormo em equídeos

Detalhes bibliográficos
Ano de defesa: 2010
Autor(a) principal: SILVA, Karla Patrícia Chaves da lattes
Orientador(a): MOTA, Rinaldo Aparecido
Banca de defesa: SAUKAS, Tomoe Noda, COSTA, Mateus Matiuzzi da, PINHEIRO JÚNIOR, José Wilton, SILVA, Leonildo Bento Galiza da
Tipo de documento: Tese
Tipo de acesso: Acesso aberto
Idioma: por
Instituição de defesa: Universidade Federal Rural de Pernambuco
Programa de Pós-Graduação: Programa de Pós-Graduação em Ciência Veterinária
Departamento: Departamento de Medicina Veterinária
País: Brasil
Palavras-chave em Português:
Palavras-chave em Inglês:
Área do conhecimento CNPq:
Link de acesso: http://www.tede2.ufrpe.br:8080/tede2/handle/tede2/5700
Resumo: The objective this study produce and evaluate PPD-mallein with potential for application in the diagnosis of glanders from of the the production and purification of immunogenic proteins of B. mallei isolates from equines with glanders in Brazil. Were isolated and characterized phenotypically and by detecting the activity of proteases, polyphenol oxidases, esterases and determination of the resistance profile in vitro. Samples of B. mallei studied were susceptible to antimicrobial gentamicin, ciprofloxacin, norfloxacin, doxycycline and enrofloxacin and resistant to trimethoprim / sulfamethoxazole, amoxicillin and ampicillin. Was observed the activity of proteases, absence esterases polyphenyloxidase and bacterial growth resulted in toxic metabolites. For the production and purification of the protein partially mallein were used two strains of B. mallei Brazilian already characterized phenotypically and genotypically. Was inoculated broth samples Dorset-Henley to metabolize and get the bacterial proteins. Proteins were separated and precipitated with trichloroacetic acid and ammonium sulphate. The PPD-mallein were concentrated at 1.0 mg / mL and evaluation in guinea pigs previously sensitized with bacter were effective for identifying infection in animals truly positive and exclude the animals truly negative. Histological analysis of the application site of mallein revealed the development of Type IV hypersensitivity. Was evaluated the PPD-mallein about their power in five asinines with clinical signs, with bacteriological diagnosis and positive serology for glanders and five asinines negative in serology and bacteriology. The animals were tested according to the criteria established by paragraph IN nº24 regarding the diagnosis of glanders. After 48 hours of inoculation, there was swelling in the area of injection, presence of ocular discharge and tearing confirming the diagnosis of glanders. The other seronegative animals showed no inflammatory reaction at the site of inoculation of PPD-mallein. This immunogen produced and being tested in Brazil was effective being a new possibility for diagnosis and control of glanders in this country.