Antibodies to the hepatitis E virus (HEV) were detected by an enzyme immunoassay using synthetic HEV peptides. Positive anti-HEV results were confirmed by a neutralization assay and Western blot analysis. Anti-HEV was detected in 10 of 555 canoeists (1.8%) with regular exposure to sewage-polluted water and in 6 of 227 (2.6%) medical students with minimal exposure. The overall prevalence of 16 per 782 individuals (2.05%) suggests that HEV may be endemic in South Africa. This is confirmed by indications of infection earlier than the third decade of life, and by individuals with anti-HEV who had rarely or never been out of the country. The prevalence data suggest that regular exposure to sewage-polluted water was not a particular risk factor. None of the individuals with anti-HEV had a history of clinical hepatitis E, suggesting sporadic low level subclinical cases of infection. This is in agreement with the absence of reports on clinical cases or outbreaks of hepatitis E in South Africa.