Isohltion, Identification and Antibiotic Sensitivity of Mycoplasma Species from Commercial Poultry Flocks in the Rawalpindi Division

Abstract

Mycoplasma galisepticul11 (M. galisepticum) and Mycoplasma synoviae (M synoviae ) are important avian pathogens and cause eco nomic losses to the poultry industry. Mo lecul ar biology techniques, such as PCR, are currently used for rapid detection of these pathogens a nd adopti on of co ntrol measures of the diseases. The a im of this study was to know the incidence of M galisepticUl11 and M synoviae in local commercial poultry, to eva lu ate the efficacy of a PCR to detect the M galisepticum and M. synoviae and to analyse the antibiotic sensitivity of the M galisepticul11 iso lates. For this purpose, tracheal swabs and tracheal tissue were taken from birds with signs of res piratory distress from 424 se lected commerc ia l pou ltry £locks (broiler, laye r and breeder). These £locks were from different distri cts of Rawalpindi Division, Punj a b province, during July 20 15 to June 20 16. M gallisepticul11 was iso lated and confirmed by growth characteristi cs and PCR o n genomic DNA from the iso lates. At the same time ge nomic DNA from tissue samples and swabs taken from infected birds was extracted using PureLink Genomic DNA Extraction kit following manufacturer's protocol and PCR was performed for .tv!. galisepticum and M sy noviae using primers given in the OlE's o nline manual. The results of PCR for M. gallisepticUl11 corre lated with the results obtained by isolation of M gallisepticul11 indicating the tested PCR can be Lisee! for direct detecti on of M gallisepticum in infected birds without the need for isolation of this organism. Lastly, antibiotic sensiti v ity was performed in PPLO broth by the broth microdilution method. Out of 424 suspected cases, a tota l of 50 £locks were found positive for M. gallisepticum. Thus overa ll incidence was 11.79%. For individual bird type the (incidence of M. gallisepticum was 12.90 %, 10. 19% and 10.34% in broilers, laye rs and breeders res pective ly. Out of the 424 £locks, 90 flocks we re also screened for M synoviae through PCR on genomi c DNA from tissues and swabs and 17 ( 18.88%) £locks we re found to be positive. For individua l bird type the incide nce of M synoviae was 19.04 %, 18.18% and 19. 14% in broilers, layers and breeders respectively. Incid ence of M. gallisepticum and M synoviae was hi gher in winter season compa red to the summer season as compared to those reared in summer. Results of antibiotic se nsitivity of the 50 M galis epticum isolates revealed that they we re resistant to multiple drugs. Ty losin and Ceftifer we re fo und to be hi ghly effective drugs for the treatment of Mycoplasma gallisepticim iso lates, while Oxytetracyc line showed limited therapeutic value for treating Mycoplasmosis. E nro£loxaci ne, Co listin, Chlorotetracyc line found to be least effective against M. gallisepticum.