High richness of ungulate Sarcocystis species in intestines of the grey wolf (Canis lupus) from Lithuania.

There is a lack of studies on the richness of Sarcocystis species in naturally infected grey wolves that serve as definitive hosts of these parasites. In this work, we used a nested PCR approach to examine the intestines of 13 grey wolves from Lithuania for the presence of Sarcocystis species that use canids as definitive host and ungulates as intermediate host. Under a light microscope, Sarcocystis spp. sporocysts/oocysts were found in 12 animals (92.3%). Based on comparisons of newly acquired cytochrome c oxidase subunit I (cox1) gene nucleotide sequences, we identified a total of 12 different Sarcocystis species (S. arieticanis, S. alces, S. capracanis, S. capreolicanis, S. gracilis, S. hjorti, S. iberica, S. linearis, S. miescheriana, S. pilosa, S. tenella and S. venatoria). This is the first report identifying four of them (S. alces, S. iberica, S. pilosa and S. venatoria) in the intestines of grey wolves. Two or more parasite species (up to nine different species per animal) were detected in 76.9% of the examined samples. This shows the significant role of the grey wolf in the life cycle of Sarcocystis species in the wild.