Background: Hepatozoon canis is a haemoprotozoan in domestic and wild dogs that is transmitted by Rhipicephalus spp. A survey was carried out to identify the tick vector of H. canis in dogs in the Mashhad area, Khorasan Razavi Province from 2020 to 2021. Materials and Method: A total of 976 ixodid ticks were collected from 39 sheepdogs and 76 shelterdogs during activating seasons of ticks. Adult ticks were identified according to general identification keys. The collected female ticks were separated into 81 tick pools according to their species. The hemolymph smear was prepared from engorged and semi-engorged female ticks and stained with the Giemsa method, Then, the DNA of each tick pool was extracted using a commercial kit and analyzed by PCR. Two ixodid species, Rhipicephalus sanguineus and Rhipicephalus turanicus were identified in infested dogs. Results: The frequency of R. sanguineus and R. turanicus infestation in sheepdogs was 80.25% and 19.75 % and in shelterdogs 76.5% and 23.5%, respectively. Hepatozoon canis DNA has only been detected in 15 (18.5%) tick pool samples of R. sanguineus. The Hepatozoon oocysts were observed in two hemolymph smears of engorged female R. sanguineus that were collected from shelterdogs. Discussion: It is concluded that R. sanguineous was the dominant tick in sheepdogs and shelter dogs in the Mashhad area. Moreover, the results of the molecular and parasitological examination indicated that R. sanguineus ticks could be a vector of H. canis in dogs in Iran for the first time

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