Serum samples from 305 white-tailed deer (Odocoileus virginianus) from 14 states in the southeastern United States were examined for antibodies to Neospora caninum using a direct agglutination test. Positive agglutination titers were found in 145 (48%) of the white-tailed deer examined: 21 (7%) had titers of 1:25, 92 (30%) had titers of 1:50, and 32 (10%) had titers of > or = 1:500. These findings that antibodies to N. caninum are common in white-tailed deer support the concept that a sylvatic cycle might exist for this economically important parasite of domestic cattle.