A cross sectional survey was conducted to determine the seroprevalence of brucellosis in cattle in Bangladesh Agricultural University (BAU) Veterinary Clinics, in BAU Dairy Farm and Vabokhali from June 2008 to November 2008. A total of 200 serum samples were collected from BAU Veterinary Clinic, from BAU Dairy Farm and Vabokhali. Among the serum samples 143 sera samples were collected from BAU Veterinary Clinic, 42 serum samples from BAU Dairy Farm and 15 serum samples from Vabokhali. Sera were separated from blood samples and tested with specific Brucella abortus antigen (BAA) test and B. melitensis antigen (BMA) test. The overall seroprevalence of brucellosis in cattle was 5% in BAA and 0.5% in BMA. It was observed that, a significant higher prevalence of B. abortus was found in female than male. An insignificant higher prevalence of brucellosis was found in adult cattle (aged above 5 years), in cross breed cattle, in cattle with grazing, cattle breed by natural breeding, and in pregnant cows. Although insignificant but a higher prevalence of brucellosis was found in aged cattle than young cattle, cross bred cattle, pregnant cattle than non pregnant cattle, cattle with grazing. A higher prevalence of brucellosis was found in female cattle than male.