To determine the seroprevalence of Brucella-specific antibodies in rural and suburban communities in different provinces of Anatolia. Cross-sectional seroepidemiological study on serum samples collected in communities in two relatively developed provinces in west Anatolia with an official low prevalence of brucellosis and in one province in southeast Anatolia with a high prevalence. The seroprevalence of brucellosis in the two provinces in the west Anatolia appears to be high and ranged from 2.9 to 8.5% in Rose Bengal test and from 0 to 5.6% in Wright serum agglutination test at a titer equal or higher than 1:100. The seroprevalence in communities in the province in southeast Anatolia was lower and this might well be attributed to vaccination of livestock in the year preceding the survey. Adherence to traditional farming practices and lifestyle, and a preference for fresh dairy contribute to the high seroprevalence of brucellosis. Vaccination of livestock is of utmost importance and the consumption of fresh milk and dairy products prepared from unpasteurised milk should be halted. Better access to laboratory testing is needed for the confirmation and management of brucellosis.