Brucellosis is a bacterial zoonosis, considered an occupational disease that has high socioeconomic impact, with more than 500,000 new cases each year worldwide. In Colombia there are very few studies, temporally and geographically isolated. The population at risk are salaugtherhouse’s workers, milkers, veterinarians and bacteriologists. Traditionally, the diagnosis has been made by Rose Bengal and confirmed by competitive ELISA or pathogen isolation. The purpose of this study was the detection of antibodies to Brucella abortus in slaughterhouse’s workers and milkers in Monteria, Cordoba (Colombia) and, additionally, the two conventional methods were compared with Fluorescence Polarization. Blood samples were taken without anticoagulation from 162 workers and processed for antibodies against B. abortus with Rose Bengal, competitive ELISA and two Fluorescence Polarization protocols. A survey to determine the level of knowledge and attitudes that have volunteers about the problem was implemented. It was found that 7.41% of the samples were positive by Rose Bengal, 2.8% with protocol 1 and 15.33% with protocol 2 of fluorescence polarization. The samples that were positive with either method were confirmed by competitive ELISA. A seroprevalence of 0.7% was found. In this population there is a low seroprevalence of antibodies to B. abortus; 50.81% of the slaughterhouse workers and 47.11% of the farms workers, know the disease and assume attitudes to prevent contagion.