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The prevalence of brucellosis was determined in the ruminants (buffaloes, cattle, sheep and goats) of five different districts viz. Bagerhat, Bogra, Gaibangha, Mymensingh and Sirajgonj of Bangladesh. A total of 550 sera samples of 105 buffaloes, 188 cattle, 127 goats and 130 sheep were screened by RBT and were further confirmed with I-ELISA. A structured questionnaire was used to collect epidemiological information on the animals. The overall serological prevalence derived from the samples was 2.87% in buffaloes, 2.66% in cattle, 3.15% in goats, and 2.31% in sheep. The prevalence was relatively higher in females than that in males in cattle, goats and sheep but, an insignificantly higher prevalence was observed in males than that in females in the case of buffalo. A significant association was found between abortion or age and occurrence of brucellosis (P< 0.01). The results of the study provide (a) a comparison of the prevalence of brucellosis in different livestock species in Bangladesh,(b) constitute baseline data for further study of Brucella infections, and (c) are a starting point for the control of brucellosis.