Brucellosis remains an urgent problem to date. The preservation of brucellosis infection foci in regions with traditionally developed animal husbandry poses a danger of introducing the pathogen into safe territories, spreading among susceptible livestock and the risk of human infection. When fresh foci of brucellosis occur in an area with long-term well-being, objective data on the causative agent of infection are of great importance. The purpose of the research is to obtain an objective result of laboratory studies when diagnosing brucellosis in cattle in a fresh outbreak of infection and to evaluate the effectiveness of a set of diagnostic methods used in the work. The research was carried out in production veterinary and scientific laboratories. It was found that brucella, forming a fresh source of infection in cattle herds against the background of long-term well-being, had active antigenic properties. In 42% of infected animals (5 out of 12 animals), brucella of the epizootic strain caused the synthesis of specific immunoglobulins. Isolated brucella cultures were characterized by a weakly expressed ability to adapt to the conditions of an artificial nutrient medium and the body of laboratory animals. In the conducted studies, the probability of isolating the causative agent of brucellosis from animals that respond positively to serological examination was 20%. The objective data obtained on the properties of brucellosis pathogens circulating among susceptible animals in a fresh outbreak of infection made it possible to determine its species differences; establish factors and ways of introducing brucella, predict the course of infectious and epizootic processes, make a timely decision on carrying out health and preventive measures using effective methods and means.