Isolation and Characterization of Pasteurella multocida from Infected Animals

Abstract

Haemorrhagic septicaemia is an acute and fatal disease among cattle and buffaloes caused by Pasteurella multocida. The prevalence of this disease is 49% in animals of Pakistan. The present study was undertaken to evaluate the prevalence, multidrug resistance and the molecular heterogeneity within clinical strains of P. multocida collected from different cities of Pakistan. Tissue samples of lung and spleen of dead animals were used for diagnostics. Rapid plate screening test for Hyaluronidase of clinical Isolates showing clear and white zones proved that given isolate is pathogenic. Amongst one hundred and ninety-nine isolates, only one hundred and one isolates were found to be virulent as clinical isolate of P.  multocida. Isolates were confirmed through amplification (PCR) of specie–specific and Haemorrhagic septicaemia type B causing test.  The primer pair of specie –specific test showed a band of 300bp in all subspecies of P.  multocida whereas the primer pair of HS type B causing test showed a band of 460bp in all HS causing serotype of P. multocida. The high prevalence was found only in Karachi. Whole cell and envelope protein profile of the isolates on SDS-PAGE showed presence of common proteins of 55, 43 and 26kDa indicating no molecular heterogeneity among strains.  Representative samples of clinical Isolates from nine different cities were tested for antibiotics sensitivity test. Isolates from Karachi, Badin, Taxila and Abbotabad showed resistant against, Ciprofloxacin, Neomycin, Ofloxacin and Norfloxacin. This study concluded that organism was genetically conserved and there is no diversity within the number of samples tested.