Identification and molecular characterization of mycoplasma species from bovine lungs samples collected from slaughter house, Quetta, Balochistan, Pakistan

Bovine respiratory mycoplsamosis is a world wide problem. The aim of this study was to highlight the presence of Mycoplasma species in the lung samples from bovine. Five hundred (n=500) lungs samples of cattle slaughtered at abattoir in district Quetta, Balochistan were collected and processed for the isolation and identification of Mycoplasma species using biochemical tests and molecular (Polymerase Chain Reaction-PCR; Restriction Fragment Length Polymorphism-RFLP) techniques. Two types of Mycoplasma species were identified. Eleven (2.2%) samples were found positive for Mycoplasma mycoides subsp.capri and two samples (0.4%) were positive for Mycoplasma arginini. Acholeplasmas (1%). All the isolates were confirmed as members of Mycoplasma genus on the basis of digitonin sensitivity test. All the isolates showing the zone of inhibition less than 2mm were considered as digitonin resistant. PCR products (Amplicon) of 1500 and 574 bp were obtained for Mycoplasma “mycoides cluster and Mycoplasma “mycoides sub cluster” respectively. PCR results for the Mycoplasma mycoides subcluster was further validated by using RFLP which yielded three bands of 230bps, 178bps and 153bps specific for Mmc when digested with Vsp1. The results of the study suggested the possibility of strains crossing the species barriers.