Serological responses to Cryptosporidium-specific antigens (15/17 and 27 kDa) were compared among populations in four areas of the Czech Republic that use drinking water from clearly defined sources: (1) wells in a fractured sandstone aquifer, (2) riverbank infiltration, or (3) two different filtered and chlorinated surface waters. Among persons surveyed in the area with riverbank-infiltration water, 33% had a strong serological response to the 15/17-kDa antigen group whereas, in the other three areas, over 72% of persons had a strong response. These response differences suggest that Cryptosporidium exposures and infection were lower in the area with bank infiltration. The large percentage of the study population with a strong serological response to both antigens suggests high levels of previous infections that may have resulted in protective immunity for cryptosporidiosis. This may be one reason why no waterborne cryptosporidiosis outbreaks and few cases of cryptosporidiosis have been reported in the Czech Republic.