European grey wolves (Canis lupus) may act as reservoirs or incidental hosts for several protozoan parasites, including Babesia spp. and Hepatozoon spp., which are relevant for both wildlife and domestic animals. This study aimed to investigate the occurrence of Babesia spp. and Hepatozoon spp. in free-ranging wolves from Lithuania and to characterise the detected parasites using molecular and phylogenetic methods. Screening for the presence of protozoan pathogens was performed by testing spleen samples from 70 wolves with PCR. Overall, 59 of 70 wolves (84.3%) were infected with at least one pathogen. Hepatozoon canis was highly prevalent, detected in 58 wolves (82.9%), while Babesia canis was found in 21 individuals (30.0%). The co-infection of B. canis and H. canis was found in 20 wolves (28.6%). This study provides the first molecular evidence of B. canis and H. canis infections in wolves from Lithuania, representing, to the best of our knowledge, the northernmost record of these pathogens in European wolf populations.

Keywords: grey wolves; Canis lupus; Hepatozoon canis; Babesia canis; Lithuania