Babesiosis is an important tickborne disease in the sheep of Iran. OBJECTIVES: A molecular study was carried out in North Khorasan province, Iran in 2010-2011, designed to identify Babesia spp. infection of both sheep and ticks. METHODS: Ninety sheep from different flocks were clinically examined and blood samples were collected with ixodid  ticks . The collected ticks were separated into 82 tick pools and the salivary glands were dissected out in 0.85% (w/v) saline under a stereomicroscope. The blood and the salivary glands were examined using semi-nested PCR. RESULTS: Piroplasm infection  was detected in 37 blood smears using microcopic examination while 80 blood samples were piroplasm positive in the first round of semi-nested PCR  and  Babesia ovis was  only detected in 6 (6.6%) of  positive samples in the second round of semi-nested PCR. Of the 434 ticks that were collected, the most prevalent species was Rhipicephalus turanicus (69.3%) followed by Hyalomma marginatum turanicum (18.4%), Dermacentor marginatus (6.4%) and Rhipicephalus bursa (5.7%). One pool of H. m. turanicum salivary glands and one pool of  R.turanicus were infected with B. ovis. CONCLUSIONS: Based on these results, it is concluded that B.ovis has a low prevalence among the sheep of North Khorasan province and H. m. turanicum and R. turanicus may be the vectors of B.ovis in this area.