A total of 1,270 nursing students of Nagasaki University School of Health Sciences were examined for serum HBsAg and anti-HBs at their first year between 1990 and 2006. The prevalence of HBsAg was 0.39% (5/1,270); 0.52% (4/762) in 1990-1999, 0.27% (1/368) in 2000-2004, and 0% (0/140) in 2005-2006. The prevalence of anti-HBs was 1.81% (23/1,270); 1.97% (15/762) in 1990-1999, 0.82% (3/368) in 2000-2004, and 3.57% (5/140) in 2005-2006. A total of 650 students who were negative for HBsAg and anti-HBs in 1998-2006 received a primary series of hepatitis B vaccinations. Subsequently, 98.2% (638/650) acquired anti-HBs. The median titer of anti-HBs concentrations at 4 weeks after primary vaccination was 2,145.0 mIU/mL. Eleven among 12 students who did not acquire anti-HBs after a primary vaccination received a second series of vaccinations, after which eight (72.7%) acquired anti-HBs. Overall 99.4% (646/650) of the students vaccinated acquired anti-HBs. Among 286 students who acquired anti-HBs after the primary series of vaccinations, 17 (5.9%) became negative for anti-HBs in their third year. Nursing students should be educated about the risks for and prevention of blood borne infections, including the need to be vaccinated against hepatitis B. Moreover, nursing students who do not acquire anti-HBs after primary vaccination should receive a 3-dose revaccination series.