Koshi Tappu Wildlife Reserve is the home for only remaining population of wild buffaloes in Nepal. It shares habitat with domestic buffaloes and human, which has increased the risk of persistence of diseases like Bovine Tuberculosis(BTB). A study was done in Buffer Zone of this Wildlife Reserve to study frequency of BTB and risk factors associated with it. Tuberculin testing was conducted among selected domestic buffaloes using single intradermal test (SIDT); 11.71% were found positive to BTB. Several factors like age of animal, lactation and period of domestic buffaloes grazing inside the reserve were significantly associated with BTB infection. Although 24.5% of interviewed households had at least one human tuberculosis case in the family, no statistically significant association was observed between diseased buffalo and human tuberculosis in the household. Only few people in the area had knowledge of BTB, and they were not conscious of its transmission between livestock, human and wild animals. This study emphasizes the risk of disease transmission in wildlife-livestock interface which is useful reference for the professionals working in wildlife reserves.