Objective: to estimate the seroprevalence of arboviruses, syphilis, HIV, and hepatitis B (HBV) in Healthcare Workers (HCW). Methods: a survey among HCW among was randomly selected in a city in Bahia-Brazil. The research used a structured questionnaire, which collected sociodemographic, occupational, and health information in 2019. Rapid immunochromatographic tests were used to track infections. Thus, positive results for arboviruses, syphilis, HIV, and HBV indicated seropositivity for the respective infection. In addition, simultaneous detection of antibodies for zika (ZIKV) and dengue (DENV) viruses indicated seropositivity for flavivirus infection. Results: a total of 453 HCW were included, 82.8% were women. Most HCW (55.1%) reported contact with biological material; 5.2% reported injuries with biological material. The seroprevalence was 34.7% (95%CI:30.1-39.4) for dengue, 1.7% (95%CI:0.7-3.4) for zika, 9.9% (95%CI:7.2-13.2) for chikungunya, and 39.9% (95%CI:35.2-44.7) for flavivirus (ZIKV+DENV); with 21.9% (95%CI:18.1-26.2) being negative to all arboviruses. Seropositivity to arboviruses increased with age and was higher among endemic disease control agents. Three HCW tested positive for HBV (HBsAg); no HIV cases were detected. Conclusion: a high percentage of HCW was exposed to infectious agents (contact with biological material and/or exposure to arboviruses). Among the necessary conditions for preventing infections in the work environment, the following stand out: monitoring of infectious diseases among HCW, surveillance of work environments, and measures to control occupational exposures, such as the availability of repellents.