| Summary: | Tick-borne protozoans of the genus <i>Hepatozoon</i> have been associated with infections of domestic and wild animals over the world. The occurrence of these apicomplexan agents in Sardinia has been poorly explored so far. In this study, the occurrence of <i>Hepatozoon</i> spp. has been investigated in domestic dogs from nine cities of Sardinia, Italy. Blood samples from each dog were collected and tested molecularly for the presence of <i>Hepatozoon</i> and <i>Babesia/Theileria</i> DNAs. Out of fifty-one dogs, nine were positive for <i>Hepatozoon</i> species based on the molecular detection of the parasite in blood samples. The phylogenetic relationships of strains detected here were also established. The PCR for amplification of the 18S rRNA fragment gene of <i>Babesia</i>/<i>Theileria</i> spp. did not give amplicons in any of the analyzed samples. Our results report the first molecular confirmation of <i>Hepatozoon canis</i> in Sardinian pet dogs and contribute to better understand the presence of these protozoans on the island. This study highlights the importance of recognizing and predicting the risk levels for the canine population, thus increasing the development of specific control measures. Also, since the distribution of hepatozoonosis is closely related to that of the definitive tick host, <i>Rhipicephalus sanguineus</i>, more accurate studies on <i>Rhipicephalus</i> ticks will be needed due to increasing the epidemiological knowledge of <i>Hepatozoon</i> species on the island.
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