Identification of Hepatozoon and Cercopithifilaria in ticks infesting dogs of different regions of Iran

Following previous detection of Hepatozoon canis and Cercopithifilaria bainae in dogs from Iran, this study aimed to identify potential tick vectors collected from dogs from five provinces of Iran. From October 2018 to September 2021, a total of 222 owned and 541 sheltered dogs from Hamedan (n=305), Kermanshah (n = 227), Yazd (n =80), Khouzestan (n =71), and Mazandaran (n =80) were examined for ticks. Collected ticks (n =3339) were identified morphologically, and adults were screened for the detection of Hepatozoon and Cercopithifilaria either with conventional PCR (n=119 ticks), tick dissection (n=106), or microscopical examination after fixation in formalin (n =102). A total of 169 dogs (22.1%) were infested with ticks. Tick infestation rate varied in different provinces (i.e., 32.4% in Hamedan, 27.3% in Kermanshah, 10.0% in Yazd), but no ticks were collected in dogs from Mazandaran and Khouzestan. The majority of male ticks were identified as Rhipicephalus sanguineus sensu lato (89.3%) and Rhipicephalus turanicus, but one dog from Kermanshah was infested with Hyalomma asiaticum (1 male and 1 female) and Hyalomma excavatum (1 male). At the molecular examination, Cercopithifilaria was detected in 14.28%, Hepatozoon in 6.7% ticks, and both parasites in 1.68% of Rhipicephalus ticks. Parasites were not observed at tick dissection and histological examinations. Nucleotide sequencing and phylogenetic analyses revealed C. bainae and H. canis in R. sanguineus sensu lato and R. turanicus ticks. Data suggests that both tick species above may play a role in the epidemiology of Cercopithifilaria and Hepatozoon in Iran.