Protozoan parasites of the genus Hepatozoon, transmitted by ticks, are known to infect both domestic and wild animals worldwide. However, their distribution and occurrence in Sardinia remain largely unexplored. The present study investigated the presence of Hepatozoon spp. in domestic dogs across nine Sardinian localities. Blood samples were obtained from all animals and analyzed molecularly to detect DNA from Hepatozoon and Babesia/Theileria species. Among the 51 tested dogs, nine were positive for Hepatozoon spp., as confirmed by molecular identification of the parasite. Phylogenetic analysis was conducted to determine the genetic relationships of the identified strains. PCR amplification targeting the 18S rRNA gene of Babesia/Theileria spp. Yielded no positive results in any sample. This investigation provides the first molecular evidence of Hepatozoon canis infection in domestic dogs from Sardinia and expands the understanding of the distribution of these protozoan pathogens on the island. The findings underscore the importance of assessing and anticipating infection risks in the canine population to support the implementation of effective control measures. Moreover, as the spread of hepatozoonosis is closely linked to its definitive vector, Rhipicephalus sanguineus, further focused research on Rhipicephalus ticks is essential to enhance the epidemiological understanding of Hepatozoon spp. in this region.
