In the present study, Indirect Fluorescent antibody test (iFAT) has been used as the "screening test" for the detection of MAP bacilli in the milk samples of lactating domestic livestock. A total of 372 milk samples from lactating animals were screened by iFAT and results were compared with microscopy of milk samples and milk ELISA test. Comparative analysis of the results of three tests showed that iFAT had fairly good sensitivity (84.7%) and specificity (90.4%), with respect to ZN staining (microscopy) with kappa value of 0.735 and "good" strength of agreement. Similar comparison with milk ELISA test revealed, sensitivity (73.3%) and specificity of (72.6%) with kappa value of 0.443 and strength of agreement was moderate. Lower or higher difference in sensitivity and specificity of iFAT with respect to ZN staining and milk ELISA may be due to the difference in detection "target" of the test i.e., antigen or antibody. It was concluded that iFAT was a reliable and sensitive diagnostic test for the detection of MAP in milk and can also be used for the "mass screening" of the milk samples.

Present study was aimed to estimate the bio-incidence of Bovine Johne"s disease in native population of buffaloes (Murrah, Zaffarabadi and Bhadawari) in dairy farms and farmer"s herds located in Central and North India. Total 156 serum samples were screened from a dairy herd in Kiratpur (Malwa region), and 41.0% buffaloes were positive for Mycobacterium avium subsp. paratuberculosis (MAP) infection using "Indigenous ELISA kit". In another study, 101 young males (farmer"s herds) from the native tract (Jind / Rohtak, Haryana) of Murrah breed were screened and 80.1% were reported positive by kit. In a farm herd of Bhadawari buffaloes in its native tract (Etawah district in Uttar Pradesh) and dairy herd of Murrah buffaloes procured from native tract and kept in Mathura district of Uttar Pradesh, In the two farms 20.0% buffaloes were screened and 71.4% were positive by fecal Microscopy. Study reported high bio-incidence of BJD in native breeds of buffaloes in Malwa region of Central India, Etawah and Mathura (Uttar Pradesh) and Jind / Rohtak (Haryana) regions of North India, despite high slaughter rate, for meat harvesting. Goat based 'Indigenous ELISA kit' was employed for screening since commercial kits were costly for the screening of buffaloes. Moreover Imported 'ELISA kits' were based on PPA from 'Cattle Type' biotype', a different strain from other geographical regions / country. "Indigenous ELISA kit" using native 'Indian Bison type' biotype was efficient, specific and cost effective 'mass screening' test to estimate bio-incidence of BJD in large population of buffaloes at National level.

Paneer (fresh cottage cheese) samples collected from local dairy shops in rural (Farah), peri-urban and urban (Mathura and Agra) regions of South Uttar Pradesh were screened by six tests combinations to estimate the rate of 'bio-contamination' and 'bio-typing' of MAP (paraTB bacilli). Of 55 paneer samples screened, 54.5, 32.7, 54.5, 74.5, 50.9 and 63.6% were positive by microscopy, IS900 PCR, indigenous ELISA kit, Dot-ELISA, Latex Agglutination test and indirect Fluorescent Antibody tests, respectively. Except 2 (3.6%), all the positive samples were detected by minimum two tests. Of the six test combinations used and compared, microscopy (AFB vs iFAT and IS900 PCR were best combinations for the detection of MAP in fresh cottage cheese (paneer) samples. High rate of bio-contamination of paneer sold by the local dairy shops was due to the use of milk supplies contaminated with high bio-load of MAP. Milk supplies were sourced from domestic livestock belonging to local rural areas, where animal health care facilities and knowledge on bio-contamination of MAP were non-existent. Therefore, in order to produce clean milk, free of MAP bacilli it is absolutely essential to control infection in the domestic livestock.