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

Résumé

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).


Sujets)
Animaux , Bovins , Lapins , Infections à Corynebacterium , Corynebacterium pseudotuberculosis/enzymologie , Corynebacterium pseudotuberculosis/isolement et purification , Phospholipase D/génétique , Phospholipase D/isolement et purification , Régulation de l'expression des gènes , Techniques in vitro , Lymphadénite , ARN , Buffles , Électrophorèse , Activation enzymatique , Méthodes , Lapins , Ovis
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