The article presents the results of allergic and bacteriological studies of cattle from a tuberculosis-free farm. The presence of cattle reacting to an allergen from atypical mycobacteria was established in three allergic simultaneous tests during 2019–2020. Based on the results obtained, the causative agent of tuberculosis and atypical mycobacteria were not isolated during the bacteriological examination of a biological material from animals slaughtered for diagnostic purposes (n = 17), as well as soil (n = 5) and straw (n = 3) samples. However, microorganisms of the genera Nocardia (n = 2), Rhodococcus (n = 10), and Actinomyces (n = 5) were isolated from the biomaterial according to the results of microscopy, culture and biochemical tests. These microorganisms were also isolated from the soil and straw samples. Short-term non-specific reactions in cattle to mycobacterial allergens were due to the persistence and circulation of the aforementioned microorganisms closely related to mycobacteria. It was determined that the genera Nocardia and Rhodococcus are sensitive to the 1.0%, 2.0%, 3.0% solutions of glutaraldehyde, formaldehyde and sodium hydroxide. Thus, it is necessary to take into account the epizootic situation as to the presence of nocardioform microorganisms in the herd during routine allergic studies, as well as in case of differentiation between specific reactions and paraallergic and pseudoallergic ones. It is necessary to carry out comprehensive systematic studies of livestock and feed quality assessment. For the purpose of disinfection, glutaraldehyde and formaldehyde at a concentration of 1.0% are effective in destroying microorganisms of the genera Nocardia and Rhodococcus with 3-hour exposure or more.