Abstract
This study investigated the pathological and parasitological causes of death in two two-toed sloths (Choloepus didactylus) housed at a zoo in Hokkaido, Japan. Genomic DNA was extracted from the clotted blood of these animals and subsequently examined by nested PCR assays targeting the 18S rRNA and β-tubulin genes. The nucleotide sequences of these genes were determined and a phylogenetic analysis of the 18S rRNA gene was performed. One sample was detected positive in nested PCR. Sequencing analysis revealed that the detected parasite was closely related to the order Piroplasmida and was subsequently designated as Piroplasmida sp. Choloepus didactylus-1 (Piroplasmida sp. CD-1).