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Braz. j. microbiol ; 43(2): 552-559, Apr.-June 2012. ilus, tab
Artigo em Inglês | LILACS | ID: lil-644470

RESUMO

Fourteen isolates of Corynebacteruim pseudotuberculosis of them 7 were isolated from sheep with Caseous Lymphadenitis "biotype 1" and 7 isolated from buffaloes with Oedematous Skin Disease "biotype 2". All isolates were identified by standard microbiological techniques and by polymerase chain reaction targeting, 16S rRNA and phospholipase D genes. Synergistic haemolytic titers of all isolates were assayed by plate technique. The presences of phospholipase D gene in supernatants of all isolates were performed by sodium dodecyl sulfate polyacrylamide gel electrophoresis immunoblot technique by using hyperimmune serum raised in rabbit immunized with recombinant phospholipase D gene antigen. The concentration of phospholipase D gene was assayed by scanning the bound phospholipase D gene with specific antibodies that appeared at 31.5 kDa. Results presented that there is no correlation between titer of Synergistic haemolytic activity and the actual phospholipase D genes concentration in culture supernatants. Also results presented that Synergistic haemolytic activity and phospholipase D genes produced by biotype 2 (buffalo isolates) was generally higher than those by biotype 1(sheep isolates).


Assuntos
Animais , Bovinos , Coelhos , Infecções por Corynebacterium , Corynebacterium pseudotuberculosis/enzimologia , Corynebacterium pseudotuberculosis/isolamento & purificação , Fosfolipase D/genética , Fosfolipase D/isolamento & purificação , Regulação da Expressão Gênica , Técnicas In Vitro , Linfadenite , RNA , Búfalos , Eletroforese , Ativação Enzimática , Métodos , Coelhos , Ovinos
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