Q fever is an emerging health problem in both humans and animals. To estimate the prevalence of Coxiella burnetii (C. burnetii) antibodies in the Estonian population, we analyzed plasma samples from 1000 individuals representing the general population and 556 individual serum samples from five population groups potentially at a higher risk (veterinary professionals, dairy cattle, beef cattle, and small ruminant stockbreeders and hunters). Additionally, 118 dairy cow bulk tank milk samples were analyzed to establish the infection status of the dairy cattle herds and the participating dairy cattle keepers. Questionnaires were used to find the potential risk factors of exposure. The effects of different variables were evaluated using binary logistic regression analysis and mixed-effects logistic analysis. The prevalence in veterinary professionals (9.62%; p = 0.003) and dairy cattle farmers (7.73%; p = 0.047) was significantly higher than in the general population (3.9%). Contact with production animals in veterinary practice and being a dairy stockbreeder in C. burnetii positive farms were risk factors for testing C. burnetii seropositive (p = 0.038 and p = 0.019, respectively). Results suggest that C. burnetii is present in Estonia and the increased risk of infection in humans is associated with farm animal contact.