Lyme borreliosis (Lyme disease) caused by the Borrelia burgdorferi sensu lato spirochete is the most common tick-borne infection manifested by a wide spectrum of clinical symptoms. In Poland, the preventive health care does not comprise individual farmers as it is practiced in foresters. The objective of this study was to evaluate the exposure of Polish farmers to infection with B. burgdorferi, based on serological screening test and epidemiological investigation. A total of 3,597 farmers were examined for the presence of B. burgdorferi antibodies, as well as interviewed regarding exposure to ticks and prophylaxis of tick-borne diseases. The prevalence varied between 18.2 and 50.7 % suggesting a focal occurrence of borreliosis. A significant increase in the frequency of positive reactions in the oldest age ranges was observed, equaling 30.9 % in the range of 60-69 years and 53.6 % in the range of 80-91 years. The prevalence of the anti-B. burgdorferi antibodies of IgG class (14.7 %) was similar to that of IgM class (16.0 %). Seroreactivity to B. burgdorferi antigen was significantly higher in the group of farmers exposed to repeated tick bites. Significant relationships were also found between some other risk factors and occurrence of seropositive reactions to B. burgdorferi. To the best of our knowledge, this is the first study concerning seroprevalence to B. burgdorferi carried out on such a large group of farmers. Results indicate a high risk of B. burgdorferi infection among Polish farmers and associations between some risk factors and the presence of seropositive reactions.