Toxoplasma gondii is the agent of toxoplasmosis, which is common to both humans and cattle. That is why the goal of work was to study the spreading of the agent of toxoplasmosis among cattle in certain regions of Ukraine, which in turn will make it possible to predict the appearance of the disease among people and prevent the development of its clinical signs. Whole blood or serum samples of cattle of different ages and sexes from farms in Kyiv, Zhytomyr, Khmelnytskyi and Lviv regions of Ukraine were sent to the laboratory for research. The presence of total antibodies to T. gondii was determined in the serum of the animals. To do this, we used the method of enzyme-linked immunosorbent assay using test kits from different manufacturers in accordance with the guidelines. In general, the prevalence of T. gondii among cattle in Kyiv, Zhytomyr, Khmelnytskyi and Lviv regions of Ukraine according to the results of studies was 10.3% (95% confidence interval: 6.7-15.1). At the same time, the study of animals was carried out in two stages, in 2014 and 2018, and the prevalence did not differ significantly. Thus, in 2014, according to the results of research, a positive reaction to the agent of toxoplasmosis was detected in 10 samples or 13.9% (95% confidence interval: 7.3- 23.4), while in 2018 this figure was 11 samples or 8.4% (95% confidence interval: 4.5-14.1) of the studied animals. Also, in 2014 and 2018 studies found that the highest level of seroprevalence of the agent of toxoplasmosis in both years of research was registered in cattle from farms of Zhytomyr and Kyiv regions was 11.9% (95% confidence interval: 4.5-24.4) and 11% (95% confidence interval: 5.9-18.3), respectively. During the conducted research, an increase in the prevalence of T. gondii with the age of animals was statistically significant. Among animals under six months of age, of the 65 serum samples tested, 3 tested positively to the agent (4.6; 95% confidence interval: 1.2-12.1), while in the group of animals older than three years (49 cows) 10 samples were positively responded (20.4; 95% confidence interval: 10.9- 33.4) compared to the previous group of animals, the P-value was 0.0114. So, the data obtained indicate the presence of the agent of toxoplasmosis among cattle in Ukraine, which requires further study and development of approaches to prevent human infection when eating meat and dairy products of positively reacting animals