Molecular Characterization of Sarcocystis Sporocysts in Intestines of Bobcat (Lynx rufus) from Minnesota

Sarcocystis species are heteroxenous protozoan parasites of major veterinary and public health relevance, yet the role of wild felids in their natural transmission cycles remains poorly understood. In this study, we investigated Sarcocystis infections in 189 bobcats (Lynx rufus) harvested from Minnesota during 2024-2025. Intestinal homogenates of 179 were examined microscopically, and Sarcocystis sporocysts were found in 38 bobcats (21.2%). DNA was extracted from all 38 positive samples, of which 19 were PCR-positive, and 14 yielded high-quality sequences for molecular characterization using partial 18S rRNA, 28S rRNA, and mitochondrial COI markers. Molecular results confirmed the presence of Sarcocystis bovifelis and Sarcocystis svanai-like. Due to inconsistent amplification and partial overlap among nuclear markers, only COI was suitable for phylogenetic analysis. The COI-based phylogenetic tree resolved strongly supported clades corresponding to these species (bootstrap 99-100%). These findings provide strong molecular evidence that bobcats in Minnesota serve as natural definitive hosts for S. bovifelis and S. svanai-like.