Outer Membrane Protein (OMP) profiles of pasteurella multocida isolates associated with haemorrhagic septicaemia by SDS-PAGE and western blot analysis

Abstract

Haemorrhagic Septicaemia (HS) is an acute fatal septicaemic disease of cattle and buffaloes. It is caused by Pasteurella multocida serotype B:2. The disease is of great economic importance in India mainly due to the high mortality in susceptible populations. Bacterial Outer Membrane Proteins (OMPs) play an important role in infectious process of many bacteria. In the present study, OMP of P. multocida (serogroup B) field isolates (n = 12) and a vaccine strain (P-52) were extracted by a sarkosyl method and characterized using sodium dodecyl sulphate-polyacrylamide gel electrophoresis (SDS-PAGE) and immunoblotting. A total of 9-14 different polypeptide bands were observed with approximate molecular weight ranging from 16-123 kDa. On the basis of stain intensity, 32 kDa protein appeared to be the major protein band followed by 37, 26, 29, 89 and 72 kDa bands. Two protein bands of 123 and 46 kDa were present only in vaccine strain. Protein band of 69 kDa was present only in four of the field isolates and vaccine strain whereas OMP with 43 kDa was detected in only four of the field isolates and absent in remaining field isolates and vaccine strain. Immunoblotting identified 32, 37, 72 and 89 kDa protein bands as immunogenic OMPs. It can, therefore, be concluded that outer membrane proteins play a significant role in the pathogenesis of pasteurellosis. Several OMPs act as immunogens for which the antibodies produced against them demonstrate a strong protective action and such OMP antigens may be used as component of subunit vaccines. © 2014 Academic Journals Inc.