Bio-typing of Mycobacterium avium subspecies paratuberculosis isolates recovered from the Himalayan sheep and goats

Information on bio-type profile of Mycobacterium avium subspecies paratubeculosis (MAP) in sheep flocks and goat herds of Himalayan region is not reported earlier. The aim of our study was to determine the bio-type of MAP infecting livestock of this region. A total of 71 faecal samples (sheep—57, goats—14) were screened by Ziehl–Neelsen (ZN) staining and IS 900 PCR, and then processed for culture on Herrold’s egg yolk medium (HEYM) having mycobactin J (MJ). Out of 71 faecal samples, MAP colonies were seen only in four samples (sheep—3 and goat—1). Isolates were confirmed as MAP on the basis of slow growth, acid fastness, MJ dependency, IS 900 and IS 1311 PCR. All the IS 900 and IS 1311 PCR positive samples were bio-typed by IS 1311 PCR-REA (restriction endonuclease analysis), which confirmed all four isolates as ‘bison type.’ In IS 1311 based phylogeny of MAP isolates by ClustalW method of the MegAlign program of DNASTAR Lasergene software, the four sequences of MAP isolates (NCBI sequence nos. MH988763, MH988765, MH988766 and MH988764) did not show any distinct clustering/grouping pattern. However, these four isolates showed a bit of closeness to the MAP sequences (KC990353.1 and KC990352.1) of ‘bison type’ isolated from wood bison in Canada. In conclusion, this is the first report on isolation and bio-type profile of MAP infecting sheep and goats of Himalayan region. Study will help in devising prevention and control strategies against spread of MAP infection in livestock population of Himalayan region.