Detection of Theileria and Babesia species in sheep and goats with molecular methods in the Black Sea Region

This study was carried out to investigate presence and distribution of Theileria and Babesia species with microscopic examination and reverse line blotting (RLB) techniques in sheep and goats in the Black Sea region of Türkiye and to determine the tick species of these animals in the region. For this purpose 1128 blood samples (869 sheep and 259 goat) were collected from sheep and goats by using the vacutained tubes containing EDTA in different provinces of various cities in the region in 2010 and 2011 years. A total of 2608 animals (2161 sheep and 447 goats) including blood samples collected sheep and goats were examined for tick infestation.Smears were prepared from the blood samples and stained with Giemsa and examined under the light microscope for Theileria and Babesia piroplasms. The genomic DNAs were extracted from blood samples. The length of 360-430 bp fragment in the variable V4 region of 18S SSU rRNA gene of Theileria and Babesia species was amplified using the gDNAs. The PCR products were hybridized to the membrane connected species spesific probes. Ticks were examined for identification under the stereo microscope.A total of 38 animals (3.37%) including 34 sheep (3.91%) and 4 goats (1.54%) were found to be positive for Theileria spp. piroplasms in microscopic examination of smears while Babesia spp. piroplasm could not detected.Infection rates were 34.64% in sheep, 10.04% in goats and totally 28.99% for T. ovis while 0.58% in sheep and totally 0.44% for B. ovis. However, Theileria sp. OT3 was detected in 2.65% of sheep and 2.04% of all animals, besides Theileria sp. MK had a 0.58% prevalance in sheep, 0.77% in goats and totally 0.62% with RLB. Although T. ovis and Theileria sp MK were determined in both sheep and goats, B. ovis and Theileria sp. OT3 were observed only in the sheep.A total of 812 animals (31.13%) that included 665 sheep (30.77%) and 147 goats (32.89%) were found to be infested with ticks. Totally 2797 ticks (2263 from sheep and 534 from goats) were collected. Ticks were identified as R. turanicus (%28,64), H. parva (%22,60), R. bursa (%18,27), D. marginatus (%16,55), R. sanguineus (%3,32), I. ricinus (%2,47), H. punctata (%2,36), H. marginatum (%2,22), H. sulcata (%1,39), H. excavatum (%1,18), H. concinna (%0,54) and H. detritum (%0,46). Seasonal distribution of identified ticks were detected that Rhipicephalus and Hyalomma present in spring and summer, Haemaphysalis in winter, spring and autumn, Dermacentor and Ixodes in all seasons.