Brucellosis remains a major zoonosis worldwide. This study was intended to estimate Brucellosis prevalence among people living and working in a high stocking density area (Mahlab 2) and risk factors. A cross-sectional study was done among randomly selected males and females in the area of Mahlab 2. Data was collected using questionnaires and laboratory investigations. Blood samples were collected from participants. The Rose Bengal Plate (RBP) test was used for the detection of Brucella antibodies in serum samples and a modified Rose Bengal Plate (mRBP) test was used for confirmation. A total of 69 participants were enrolled in this study. The majority of those participants were between 18–29 years of age. The seropositivity of patients’ blood samples using these tests, was 28.4%. Socio-demographic characteristics, occupation, clinical signs of disease, types, and size of herds reared by animal owners, sanitary practices, and awareness about zoonosis, Brucellosis, and symptoms of Brucellosis showed no significant correlation with seropositivity among patients’ blood samples for Brucellosis. It is recommended that surveillance and extension programs should be carried out to evaluate the magnitude of Brucellosis and to increase awareness about it. For proper control and prevention to decrease the incidence in animals, hence decreasing the incidence in humans, animal vaccination is essential.