This paper presents the results of laboratory studies, from years 2008–2018, on the distribution of bovine tuberculosis among different categories of animals. Bovine tuberculosis (BTB), is an infectious disease caused by Mycobacterium bovis or Mycobacterium caprae. Since 2009, Poland is considered as officially free from this disease, however every year new outbreaks of BTB are recognized. Tuberculosis caused by bovine type of mycobacteria affects mainly domestic cattle, but almost all mammals are susceptible. In the last 10 years, many cases in wildlife and free-living animals have been reported. The Bieszczady Mountain region is area with the highest incidence rate of BTB among wild animals. BTB was there diagnosed in bison, wild boar and wolves. Due to the high virulence, bovine mycobacteria easily spread among species of animals living in the same feeding grounds. The major source is the contaminated soil in which mycobacteria can live up to several years and retain their infectivity. There is also high risk of disease transmission to the farm animals. At present, there are no regulations, concerning the epidemiological situation of BTB in wildlife and free-living animals. Despite this, the National Veterinary Research Institute continuously perform laboratory studies on specimens from selectively shot animals or carcasses