Objective To investigate the brucellosis infection and risk factors among occupational populations in Zhejiang province, and to provide scientific basis for preventive measures. Methods Occupational populations engaged in cultivation, transportation, trading, veterinarian services, fur making, marine fishing from October 2015 to March 2016 were selected for study. Rose bengal plate test (RBPT) and Serum agglutination test (SAT) were used for serological test. A case-control study was conducted to analyze the risk factors through logistic regression analysis. Results Among all 3 410 respondents, 105 were infected with brucellosis, with the infection rate of 3.08%. The difference of infection rate between male (3.10%) and female (3.02%) had no statistical significance (χ²=0.010, P=0.970). Multivariate logistic regression analysis showed that no wearing gloves and hand wound were found to be associated with the incidence of brucellosis. Conclusion Risk factors for the incidence of brucellosis in Zhejiang province were no wearing gloves and hand wound. Health education and intervention should be strengthened to improve the protection and awareness of brucellosis in occupational populations.