Introduction: The World Health Organization (WHO) describes One Health as an approach that integrates human, animal, and ecosystem health in order to control diseases. The Mycobacterium tuberculosis complex (MTBC) includes M. tuberculosis, responsible for human tuberculosis, and M. bovis/caprae - primarily cause bovine tuberculosis, and can also infect humans (zoonotic tuberculosis). The Interferon-Gamma Release Assay used in humans demonstrates immune sensitization to infection with MTBC. The IFN-γ is used in animals to determine sensitization directed against M. bovis/caprae. The test is usually performed together with other elements of bovine tuberculosis control programmes, such as tuberculin skin tests. The aim of this study was to evaluate the usefulness of the interferon-gamma release assay (IFN-γ) for the detection of bovine tuberculosis in selected animal species and to assess its significance for public health within the One Health framework. Methods: The IFN-γ test was used in this study. The test is adequate for intended use according to WOAH Terrestrial Manual it’s approved for use in cattle, goats, and camelids. Bison are phylogenetically closely related to cattle, and evidence indicates that this assay, is also applicable to bison, as supported by the obtained results. Results: In 2025, a total of 68 animals (including alpacas, European bison, cattle, and camels) were tested for IFN-γ release using the Bovigam test (Thermo Fisher Scientific). Among the tested samples, six European bison originating from a showing pen in the Podkarpackie Province and one cow from the Zachodniopomorskie Province tested positive for bovine tuberculosis, representing 8.8% of positive samples (6/68). Conclusions: The results indicate that this test is adequate and suitable for the early detection of bovine tuberculosis in cattle and other described animal species. After receiving positive results, appropriate decisions regarding animal management can be made in order to reduce the potential risk of infection transmission to humans.