The first published tularemia epidemic in Turkey had been reported in 1936 from Luleburgaz (located in European part-Thrace region-of Turkey), and the second was in 1945 again in the same province. Following a long period of time without any tularemia report from Thrace region, in 2005 another epidemic occurred in a village of Edirne, another province located in the same region. Since there is presumptive evidence of circulation of the infectious agent, Francisella tularensis in Thrace region of Turkey, a large scale seroepidemiological study is needed. In this study, the presence of antibodies against F. tularensis in 1782 subjects, choosen by "thirty cluster" method, inhabiting in 90 different villages of Edirne, Kirklareli, and Tekirdag provinces in Thrace Region, were investigated. The subjects were included to the study on the basis of volunteering (74.3% were male; mean age: 46 years; age range: 6-92 years) and demographical characteristics and their possible risky behaviours were recorded in a questionnaire form. Antibodies specific for F. tularensis were screened by microagglutination test, and were found positive in five (0.3%) of the subjects between the titers of 1/20- 1/160. All of the seropositive subjects were adult males (ages between 22-74 years); three were living in the two villages of Kirklareli, while the others were from the villages of Tekirdag and Edirne. Rose Bengal test was also found positive in three of the seropositive subjects, and with the thought of a probable cross reaction they were taken into an advanced investigation for brucellosis. The risk evaluation revealed that male gender, being together with livestock and exposure to ticks were the major risk factors. Since the data of this study indicated that F. tularensis is in circulation in Thrace Region, the educational programmes for both the healthcare workers and inhabitants of this region should be attempted for the prevention of a possible epidemic.