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Brucellosis is a neglected endemic zoonosis in Bangladesh and has a significant impact on public health and animal welfare of dairy farming as well as dairy farm economics. A cross-sectional study was conducted to evaluate the seroprevalence of and risk factors for brucellosis in dairy cattle in the Chittagong metropolitan area (CMA) of Chittagong, Bangladesh. We collected serum samples (n = 158) from six randomly selected dairy farms in the CMA between February and November, 2015. The Rose Bengal Plate Test (RBPT) and a competitive ELI