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• Manuscript discusses the incidence of MAP infection in diarrhoeic bovines, a question which has never been evaluated. • Sero-prevalence of JD was very high in diarrhoeic buffaloes and cattle from Haryana state of India. • Buffaloes positive for MAP infection had reduced total leukocyte count and lymphocytes indicating chronic nature of disease. • Buffalo PBMCs express TIM3. • Elucidating relative expression of TIM3 on JD positive and negative PBMCs will provide a clue on immune exhaustion in JD. Murrah buffaloes, best breed for milk production are native of Haryana state. They contributes significantly to the farmer's income, livelihood and food (milk and meat) security, in the semi-tropical regions of North India. Johne's disease though endemic in the domestic livestock of the country, but reports are not available in the buffaloes suffering from morbidity due to progressive weakness and diarrhoea. We estimated the status of JD in diarrhoeic buffaloes and cattle reporting at Veterinary Clinical Complex of Lala Lajpat Rai University of Veterinary and Animal Sciences, Hisar, Haryana, India, using conventional, serological and PCR assays.141 buffaloes suffering from chronic diarrhoea were screened to estimate sero-prevalence of MAP and 50.0 % young and 53.52 % adult animals were positive. Of 14 cattle screened, none of the young and 66.6 % adult cows were positive. In buffaloes, 66.1 and 6.77 %, fecal samples were positive in microscopy and IS 900 PCR, respectively. Sero-prevalence of JD was very high in diarrhoeic buffaloes and cattle from Haryana state of India.Buffaloes positive for Mycobacterium avium subspecies paratuberculosis (MAP) infection had reduced total leukocyte count and lymphocytes. [ABSTRACT FROM AUTHOR]