Regulations for public water systems (PWS) in the U.S. consider Cryptosporidium a microbial contaminant of surface water supplies. Ground- water is assumed free of Cryptosporidium unless surface water is entering supply wells. We determined the incidence of Cryptosporidium in PWS wells varying in surface water inﬂuence. Community and noncommunity PWS wells (n = 145) were sampled (n = 964) and analyzed for Cryptosporidium by qPCR and immunoﬂuorescence assay (IFA). Surface water inﬂuence was assessed by stable isotopes and the expert judgment of hydrogeologists using site-speciﬁc data. Fifty-eight wells (40%) and 107 samples (11%) were Cryptosporidium- positive by qPCR, and of these samples 67 were positive by IFA. Cryptosporidium concentrations measured by qPCR and IFA were signiﬁcantly
correlated (p < 0.001). Cryptosporidium incidence was not associated with surface water inﬂuence as assessed by stable isotopes or expert judgment. We successfully sequenced 45 of the 107 positive samples to identify species, including C. parvum (41), C. andersoni (2), and C. hominis (2), and the predominant subtype was C. parvum IIa A17G2R1. Assuming USA regulations for surface water-supplied PWS were applicable to the study wells, wells positive for Cryptosporidium by IFA would likely be required to add treatment. Cryptosporidium is not uncommon in groundwater, even when surface water inﬂuence is absent.