Breaking host barriers: detection of Cytauxzoon banethi in a non-felid feliform, the striped hyena (Hyaena hyaena).

Cytauxzoon banethi is one of the least characterized members of the genus Cytauxzoon and has so far been reported only from two European wildcats (Felis silvestris) in Romania. Consequently, its host range and geographic distribution remain poorly understood. In the present study, we molecularly characterized a Cytauxzoon isolate detected in a free-ranging striped hyena (Hyaena hyaena) from western Iran. Genomic DNA extracted from blood was analyzed by PCR amplification and sequencing of partial 18S rRNA and mitochondrial cytochrome b (cytb) genes. The 18S rRNA sequence was identical to published C. manul sequences and highly similar to several other Cytauxzoon species, confirming the limited species-level resolution of this conserved marker. In contrast, direct comparison of the cytb sequence with the available C. banethi reference sequence revealed only five synonymous nucleotide substitutions across 1,089 aligned positions (99.5% identity). Phylogenetic analysis of the cytb dataset placed the obtained isolate within the C. banethi lineage as the sister taxon to C. manul, supporting its identification as C. banethi. To the best of our knowledge, this represents the first molecular detection of C. banethi in a member of the family Hyaenidae, the first record in a non-felid feliform host, the first report from Iran and Asia, and the first detection of this species outside Europe. These findings expand the currently recognized host range and geographic distribution of C. banethi and emphasize the importance of multilocus molecular approaches and broader wildlife surveillance for improving our understanding of the diversity and evolution of Cytauxzoon species in general, and for better assessing the genetic diversity and taxonomic boundaries of C. banethi.