Isolation, Molecular Identification of Mycoplasma Putrefaciens Local Strains and Its Associated Pathological Lesions in Goats

Mycoplasma putrifaciens (Mp) is a dreadful pathogen that causes a severe, usually fatal disease in domestic goats known as MAKePS syndrome in small ruminants. The present lack of effective diagnostic and control measures leaves Pakistani farmers at perpetual great risk of financially disastrous MAKePS outbreaks. Thus far, very limited information is available on the detection and molecular characterization of the unique Mp strains prevalent in Pakistan. This study was proposed to apply modern molecular techniques to characterize endemic Mp isolates and their associated specific lesions in goats to devise an effective control strategies. For the collection of samples Northern areas of Pakistan were focused including Chitral, Gilgit and Swat. Samples were collected from Mycoplasma putrifaciens suspected animals (Chitral n=200, Gilgit n=200, Swat n=200). Clinical samples including nasal swab (NS) and synovial fluid (SF) from live animals (n=50), whereas pleural fluid (PF) and lungs samples from dead goats (n=50) were collected from each representative northern area of Pakistan. All samples were subjected to isolation of Mp local strain, while tissue samples were examined for pathological lesions. Culture positive PPLO broth was first confirmed for Mp by specific PCR. From Gilgit Baltistan 38.83% samples were found positive for Mp by PCR, whereas 25% and 19.38% samples from Chitral and Swat, respectively, were detected positive for Mp by generating an amplicon of 540 bp. The PCR positive PPLO broth samples were cultured on PPLO agar for isolation of Mp. A single colony with fried egg like appearance was picked and sub-cultured in PPLO broth for purification and later on confirmed with PCR. The Histopathological study of lungs showed that there was emphysema, rupture of alveoli, in bronchioles ciliated epithelium were sloughed and infiltration of leukocytes. We successfully isolated and purified Mp local strain prevalent in northern area of Pakistan. The Mp was detected prevalent in 27.76% goats of Northern regions by PCR. This study laid down a foundation to develop an effective diagnostic kit and vaccine against Mp infection in Goats.