In order to investigate the hepatitis B virus (HBV) infection profile in hemodialysis staff (n = 152) of Goiânia,Goiás, blood samples were tested for detection of HBsAg, anti-HBs and anti-HBc markers. An overall HBV infection prevalence of 24.3% (CI 95%: 17.8 - 32) was found. Multivariate analysis of risk factors showed that length of employment, history of occupational exposure and nonuse of protective equipment were significantly associated with HBV seropositivity. Of 40 staff members who were susceptible to this infection, 20 agreed to participate in the vaccination program. After three vaccine doses (Euvax-B), 18 (90%) seroconverted to anti-HBs with titers >/= 10 IU/L. These data suggest the dialytic environmental as a possible source of occupational transmission of HBV, and emphasize the need to evaluate strategies of control and prevention to be followed in these units.