To assess the risk for hepatitis A virus (HAV) infection in an occupational group potentially at risk for faeco-oral contact with very young children, a prevalence study of total anti-HAV antibodies (IgG/IgM) was conducted among 591 female employees in day nurseries in Flanders, Belgium, and in a reference group of 560 healthy female blood donors, matched for age. Analysis was also performed on formally exposed persons (n = 413) versus blood donors (n = 560). The overall prevalence of HAV markers was 48.4% (95% CI: 44.2-52.5) in exposed day nursery personnel, compared with 42.9% (95% CI: 38.7-47.0) in blood donors. The age-specific prevalence rates showed a steeper rise from the age of 30 years among the exposed employees than among the blood donors, with significantly higher prevalences between 35 and 44 years of age. The discrepancies levelled off above 60 years of age. Standardization for parenthood using logistic regression did not affect the odds ratio. These results are in line with recent findings of a higher prevalence of HAV markers among groups of workers professionally exposed to small children. Appropriate measures for the protection of these groups should be taken.