Stool samples of 8 human beings were processed for isolation of Mycobacterium avium subsp. paratuberculosis (MAP). Of these 2 were suffering from clinical bowel disorders (frequent stomach upset), weakness, weight loss and intermittent diarrhea and were positive (25.0%) for the typical MAP colonies. More than 20 colonies appeared around 26 days of incubation in one case and around 35 days in second case. Colonies were identified on the basis of morphology, media discoloration, slow growth, mycobactin J dependency and acid fastness. One person had Hb, 7.5 ESR was always more than normal and also had the history of consuming raw cow milk. Rests of the samples were negative (up to 150 days incubation), and persons were apparently healthy. Isolation of MAP bacilli from the stool of persons with clinical symptoms established the role of MAP in the pathogenesis of Crohn's disease. This study is the first report of isolation by culture of MAP from stool samples of human beings in India. However, on the basis of plate ELISA on 45 serum samples of human beings from Agra region the prevalence of Crohn's disease was 46.6% and 44.4% using Bison type and Bovine type antigens, respectively.