Cryptosporidium spp. infection in captive exotic mammals was investigated using staining and molecular biological methods. A total of 323 fecal samples from 100 mammalian species (62 Artiodactyla, 33 Rodentia, 3 Perissodactyla, and 2 Paenungultata) in 4 zoological gardens in the Czech Republic was examined. Only in a reticulated giraffe ( Giraffa camelopardalis reticulata ) sample was Cryptosporidium sp. infection detected. The partial small subunit rRNA sequence obtained from the isolate was identical to sequences of Cryptosporidium muris in rock hyrax ( Procavia capensis ) and Bactrian camel ( Camelus bactrianus ). Neonatal BALB/c mice inoculated with 1 × 10 3 fresh oocysts of the C. muris giraffe isolate did not produce a detectable infection.