Brazilian Journal of Microbiology
Print version ISSN 1517-8382
SELIM, S.A.; MOUSA, W.M.; MOHAMED, K.F. and MOUSSA, I.M.. Synergistic haemolytic activity and its correlation to phospholipase D productivity by Corynebacteruim pseudotuberculosis Egyptian isolates from sheep and buffaloes. Braz. J. Microbiol. [online]. 2012, vol.43, n.2, pp.552-559. ISSN 1517-8382. http://dx.doi.org/10.1590/S1517-83822012000200016.
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).
Keywords : Corynebacteruim pseudotuberculosis; Oedematous Skin Disease; Caseous Lymphadenitis; recombinant PLD; Synergistic haemolysis.