Cytauxzoon europaeus, Babesia vulpes, and Hepatozoon felis circulating simultaneously in a European wildcat (Felis silvestris) population in northern Spain

The Iberian population of the European wildcat (Felis silvestris) is undergoing a decline and is highly fragmented. This study aimed to determine the occurrence and identity of tick-borne parasites (piroplasms and Hepatozoon spp.) in Navarre, one of the strongholds for wildcats in Spain. DNA was extracted from spleen samples of 63 road-killed wildcats and analyzed using a suite of molecular protocols targeting the 18S rRNA and CytB gene fragments. Eighty percent of the wildcats tested positive for at least one parasite. Cytauxzoon spp. was detected in 65 % of the individuals. Molecular characterization and phylogenetic analysis confirmed the presence of Cytauxzoon europaeus in nine samples, representing the first report of this species in the Iberian Peninsula. Hepatozoon spp. was found in 46 % of the wildcats, and molecular characterization of four cases identified Hepatozoon felis. Babesia vulpes was detected in a single individual. Coinfection with Cytauxzoon sp. and Hepatozoon sp. was observed in 32 % of the wildcats, while one individual was coinfected with C. europaeus and B. vulpes. Occurrence of H. felis was higher in adult wildcats. No spatial structure or correlation with body condition was observed for any of the parasites. Given the widespread presence of Cytauxzoon and other parasites in this and other wildcat populations, their impact on wildcat health, if any, is likely to be minimal. However, wildcats appear to serve as important maintenance hosts for these parasites.