AlthoughHepatozoonspp. remains the most prevalent intracellular protozoa infecting snakes, it was reported only in a few snake species of the Colubridae family in Türkiye. Moreover, studies on these hemoparasites are not available in venomous nose-horned vipers from Türkiye. In this study, we investigatedHepatozoonspp. in three individualVipera ammodytesusing morphological and molecular methods. Our results were positive for intraerythrocyticHepatozoonspp. gamonts in all three snakes, exhibiting low parasitemia. The microscopic findings were further confirmed through molecular data. A genus-specific PCR assay targeting the 18S rRNA gene region ofHepatozoonspp., was performed using HemoF/HemoR and Hep300/Hep900 primers. The obtained sequences were concatenated and used in phylogenetic analyses in comparison with differentHepatozoonspecies. Although our (OP377741) isolate was separated into a different branch, it was clustered with the isolates ofH. massardi(KC342526),H. cevapii(KC342525), andH. annulatum(ON262426) from Brazilian snakes. Moreover, gene similarity and pair-wise distance between our isolate and otherHepatozoonspecies infecting snakes were found to be 89.30–98.63% and 0.009–0.077, respectively. Hence, we reported a new species ofHepatozoon, namelyHepatozoon viperoisp. nov. infectingV. ammodytes. Since the literature does not indicate the existence of such aHepatozoonspecies inV. ammodytesin different countries, our data may contribute to the expanding knowledge ofHepatozoonspecies in snakes, providing new insights into the biodiversity of the haemogregarine protozoan parasite.