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Paratuberculosis (ParaTB) in ruminants, commonly known as Johne's disease (JD)is caused by Mycobacterium avium subspecies paratuberculosis (MAP). Characteristic hallmark of JD is chronic inflammation of intestines. Similar infection of MAP in human population is called Crohn's disease (CD). In sub-tropical countries like India, domestic buffaloes are important source of milk and meat and vital for thenutritional security of human beings. Diarrhoea leading to weight loss and weakness invariably leads to reduced productivity, therefore huge financial losses to the livestock farmers. Evaluation of immuno-biochemical alterations and their management through appropriate therapeutic interventions would aid in preventing production losses caused by diarrhoea following MAP infection.Most of the studies were performed to understand immune-biochemical alterations in serum samples of cattle following experimental infections. But studies are limited in clinical cases of buffaloes suffering from diarrhoea and were suspected for MAP infection.We categorized diarrhoeic buffaloes brought at Veterinary Clinical Complex of Lala Lajpat Rai University of Veterinary and Animal Sciences, Hisar, for therapeutic intervention; As MAP negatives / suspected, low positive, positive and strong positive based on indigenous ELISA kit results. We further compared immuno hemato-biochemical alterations in MAP negative/suspected low positive, positive and strong positive animals.Our major findings were; significant differences observed in creatinine, globulin, total leucocytic, neutrophils and in lymphocytes numbers. Decreasing trend in leucocytes was seen from suspected to strong positive animals, which indicated the progression of disease towards chronic paratuberculosis. However, differences were not observed in the Th1 cytokine levels. • ELISA based sorting of spontaneous naturally infected diarrhoeic buffaloes as MAP negative, suspected, low, strong positive. • Plasma creatinine and globulin fraction were significantly increased in MAP strong positive diarrheic buffaloes. • Significantly reduced Total leucocyte count (TLC) in strong positive animals. • Progressive decline of TLC, neutrophils and lymphocytes from suspected to strong positive (advancement towards chronicity). [ABSTRACT FROM AUTHOR]

• 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]