Tuberculosis (TB) in humans is primarily caused by Mycobacterium tuberculosis (M. tuberculosis), with millions of infections and hundreds of thousands of deaths worldwide. It creates a substantial economic burden on the community. Unlike M. tuberculosis, Mycobacterium bovis infects cattle and causes bovine TB, also known as zoonotic TB. People can contract zoonotic TB after consumption of unpasteurized dairy products, handling the sick animals, and via occupational exposures. The association between the zoonotic TB in humans and cattle is not well known in Nepal. The study examined the associated risk factors, including exposure to infected cattle, that contribute to TB's development in human beings in Nepal. The study consists of human and animal subjects. Firstly, a retrospective matched case-control study was conducted at the National Tuberculosis Center (NTC), Bhaktapur, Nepal. A total of 290 people (equal numbers of TB cases and control subjects) were interviewed to obtain information on socio-demographic, behavioral, and occupational risks, including the history of cattle related exposures. Secondly, a cross-sectional study was performed among the cattle owned by the TB-confirmed patients. Comparative tuberculin skin test, rapid antibody test, and ELISA were used in parallel to detect M. bovis infection in cattle. The risk factors for the development of TB in humans were smokers (OR = 4.6, 95% CI: 2.1–10.0, p < 0.001), previous history of TB (OR = 7.9, 95% CI: 3.0–20.6, p < 0.001) and history of cattle exposures (OR = 3.9, 95% CI: 2.1–7.4, p = 0.001). Out of 123 cattle sampled, 12 cattle (9.76%, 95% CI: 5.37–16.76, p < 0.0001) were positive by the tuberculin test, 46 (37.4%, 95% CI: 28.97–46.62, p = 0.007) were tested positive by the rapid test, and 7 (5.7%, 95% CI: 2.52–11.80, p < 0.0001) by ELISA test. The inter-test agreement between the tuberculin and ELISA was very strong (κ = 0.72, 95% CI: 0.48–0.95, p < 0.01). This study indicates that exposure to infected cattle and socio-demographic risk factors can contribute to the development of TB in human beings.