First molecular detection of Sarcocystis spp. in intestinal homogenates of Florida panthers (Puma concolor coryi) from Florida, USA

Wild felids serve as definitive hosts for numerous Sarcocystis species infecting domestic and wild animals. In this study, intestinal homogenates from six Florida panthers ( Puma concolor coryi ) collected in Florida, USA, were examined for Sarcocystis by microscopy and molecular methods. Oocysts/sporocysts were detected microscopically in four animals following bleach digestion of intestinal homogenates. PCR amplification was successful for two of the four microscopy-positive samples. Sequence analyses of the 18S rRNA and mitochondrial cox1 loci demonstrated the presence of two genetically distinct Sarcocystis lineages. The obtained sequences showed relatively low similarity to currently available reference sequences in GenBank, preventing definitive species assignment but suggesting previously uncharacterized genetic diversity within the genus. Phylogenetic analyses placed these sequences within broader Sarcocystis clades utilizing ruminants such as goat, sheep and water buffalo as intermediate hosts. This study provides the first molecular detection of Sarcocystis in intestinal homogenates of Florida panthers and expands current knowledge of the diversity of Sarcocystis circulating in free-ranging felids in North America. Additional multilocus analyses and biological characterization will be required to determine the taxonomic identity and life cycles of these parasites.