The presence of Cryptosporidium parasites in mammals and reptiles kept at the Lisbon Zoo was investigated. A total of 274 stool samples were collected from 100 mammals and 29 reptiles. The species and genotype of the isolates identified by light microscopy were determined by nested PCR and sequence analysis of a fragment of the small subunit rRNA gene. Cryptosporidium oocysts were found in one black wildebeest (Connochaetes gnou), one Prairie bison (Bison bison bison) and in one Indian star tortoise (Geochelone elegans). The PCR and sequence analysis of these three isolates showed that those excreted by the Prairie bison were Cryptosporidium mouse genotype, those from the black wildebeest were from a new Cryptosporidium genotype and those infecting the Indian star tortoise were Cryptosporidium tortoise genotype. The present work reports a new Cryptosporidium genotype in a black wildebeest and the first finding of the Cryptosporidium mouse genotype in a ruminant.

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Feces from 34 species of ruminants housed at the Lisbon Zoo was examined for Cryptosporidium spp. oocysts. Three hundred and eighty-eight samples were analyzed. Three hundred and eighty species-specific group fecal samples were collected monthly, from September 1998 until August 1999, along with eight individual specimens from eight neonates. All samples were examined by four different techniques: microscopic observation of direct and concentrated fecal smears, staining with modified Ziehl-Nielsen, immunofluorescent assay, and immunoenzymatic assay. The prevalence of infection was 3.6%. Five neonates with diarrhea were infected. Cryptosporidial oocysts were shed more frequently during winter months. Some facilities may have permitted oocysts to remain viable, possibly contributing to cryptosporidial transmission between animals.