Objective To determine tick infestation of domestic ruminants and their infection to ovine theileriosis in northern Iran. Methods About 425 domestic ruminants in Ghaemshahr city in northern Iran were inspected for tick infestations. Twenty tick specimens (13 females and 7 males) of Rhipicephalus sanguineus (R. sanguineus), the most common tick in the study area, were tested by PCR amplification against 18s rRNA genome of Theileria spp using specie specific primers and then the PCR products were sequenced for species identification by comparison with data base available in GenBank. Results About 323 ticks were collected from 102 animals (88 sheep, 12 goats and 2 cattle). The prevalence of ticks infesting animals was R. sanguineus (82.35%), Rhipicephalus bursa (R. bursa) (0.3%), Ixodes ricinus (I. ricinus) (15.2%), Boophilus annulatus (B. annulatus) (1.2%), Haemaphysalis punctata (H. punctata) (0.3%) and Haemaphysalis numidiana (H. numidiana) (0.6%). Eleven (55%) tick specimens were PCR positive against genome of Theileria ovis (T. ovis). Sequence analysis of the PCR products confirmed presence of T. ovis in one R. sanguinus. Conclusions This is the first report of tick infection to T. ovis in Iran. Due to dominant prevalence of R. sanguineus as well as its infection to T. ovis, it is postulated this tick is the main vector of ovine theileriosis in northern Iran.