Bovine tuberculosis is a chronic disease caused by Mycobacterium bovis, a bacterium that infects a wide range of animal species, including humans. In this study, we determined the prevalence of tuberculosis in domesticated water buffalo (Bubalus bubalis) and assessed potential risk factors at four government dairy farms in Punjab, Pakistan. Tuberculin skin testing was performed on 627 water buffaloes. Risk factors such as age, body weight, milk yield, lactation status, the total number of animals, and the presence of small ruminants at the farms were recorded due to their potential association with tuberculosis transmission. The impact of M. bovis infection on complete blood cell counts (CBC) was also assessed. In total, 27 (4.3%) animals were positive for tuberculosis in the four dairy farms. Mean corpuscular hemoglobin concentrations were lower in infected animals compared to non-infected animals, whereas the number of platelets was higher in infected animals compared to non-infected animals. The results revealed that the increase in body weight, age of the animals, and water buffalo density on the farm can increase the disease occurrence.  Binary logistic regression analysis, including farm and other variables, revealed that the assessed herd-level risk factors of bovine tuberculosis transmission had a statistically non-significant association for monitoring water buffaloes. The current study had emphasized revealing different possible risk factors in native husbandry practices. The study suggests regular tuberculosis screening for proper control of M. bovis infection in water buffalo herds. This will ultimately help to further reduce the cases of zoonotic bovine tuberculosis.