Molecular Survey of Theileria and Babesia Species in Small Ruminants and Ticks from Sivas Region of Turkey

Year 2017, Volume: 7 Issue: 1, 30 - 39, 01.06.2017

Kürşat Altay Ahmet Duran Ataş Erkan Özkan

Abstract

This study was carried out to investigate the presence and distribution of Theileria and Babesia species with microscopic examination and reverse line blotting (RLB) techniques in sheep, goats and ticks from Sivas (Sivas, Kangal, Koyulhisar, Yıldızeli) of Central Anatolia region of Turkey. For this purpose 309 blood samples (206 sheep and 103 goats) and 478 tick samples that divided into 27 pools as the species were collected from sheep and goats. The total genomic DNAs were extracted from 309 blood samples and 27 tick pools were tested by RLB for Theileria and Babesia species. A total of 8 animals (2.59 %) were found to be positive for Theileria spp. piroplasms in microscopic examination of the smears. Four species or genotypes (Theileria ovis, Theileria sp. MK, Theileria sp. OT3, Babesia ovis) were detected in sheep, goats and ticks pools with RLB. 18S SSU Infection rate of T. ovis was 56.80 % in sheep, 3.88 % in goats and totally 39.16 %. Theileria sp. OT3 was detected in 4.85 % of sheep and 3.24 % of all animals, while Theileria sp. MK was detected 3.88 % in sheep and 2.59 % in all animals. rRNA gene sequences were detected for all species or genotypes. The sequences were submitted to GenBank under accession numbers; KF266694, KF266695, KF434321 and KF440691. The tick species were identified as Rhipicephalus bursa, Haemaphysalis sulcata, Hyolamma marginatum, H. detritum and Dermacentor marginatus. According to the result of tick pools analysed by RLB, T. ovis were detected in R. bursa, H. marginatum, H. detritum and Hae. sulcata while B. ovis were determined in R. bursa.

Keywords

Theileria,  Babesia,  sheep,  goat,  tick,  RLB,  Sivas

Sivas Yöresinde Koyun Keçi ve Kenelerde Theileria ve Babesia Türlerinin Moleküler Yöntemlerle Araştırılması

Abstract

Bu  çalışmada,  Sivas  yöresinde  (Merkez,  Kangal,  Koyulhisar  ve  Yıldızeli)  mikroskopik  muayene  ve reverse line blotting (RLB) metodu ile koyun, keçi ve kenelerde Theileria ve Babesia türlerinin varlığı ve yaygınlığı  araştırılmıştır.  Toplam  309  hayvandan  (206  koyun,  103  keçi),  DNA  ekstraksiyonunda kullanılmak  üzere  EDTA’lı  kan  örneği  alınmış  ve  mikroskobik  muayene  amacıyla  kan  frotileri hazırlanmıştır.  Koyun  ve  keçiler  üzerinden  toplam  478  kene  örneği  toplanmıştır.  309  kan  ve  27  kene havuzundan elde edilen DNA örnekleri RLB ile Theileria ve Babesia türleri yönünde analiz edilmiştir. Kan frotilerinin mikroskopik muayenesi sonucunda toplam 8 hayvanda (% 2,59) Theileria spp. piroplasmları belirlenmiştir.  RLB sonucuna göre Sivas yöresinde koyun, keçi ve kenelerde Theileria ovis, Theileria sp. MK, Theileria sp. OT3 ve Babesia ovis’in varlığı ortaya konulmuştur.  RLB ile incelen koyunlarda T. ovis pozitiflik oranı % 56,80, keçilerde bu oran % 3,88 toplamda ise % 39,16 olarak bulunmuştur.  Theileria sp. OT3 pozitiflik oranı koyunlarda % 4,85, toplam hayvanlarda % 3,24 bulunurken, Theileria sp. MK pozitiflik oranı koyunlarda % 3,88 ve toplam hayvanlarda % 2,59 olarak belirlenmiştir. Her bir tür ya da genotipi temsilen seçilen birer örneğin 18S SSU rRNA geni V4 çok değişken bölgesinin DNA dizilimi belirlenmiştir. Bu  dizilimler  GenBank’a  sunularak  KF266694,  KF266695,  KF434321  ve  KF440691  numaraları  alınmıştır. Kene  türlerinin  Rhipicephalus  bursa,  Haemaphysalis  sulcata,  Hyolamma  marginatum,  H.  detritum  ve Dermacentor marginatus olduğu belirlenmiştir. Kene havuzlarının RLB sonucuna göre, T. ovis R. bursa, H. marginatum, H. detritum ve Hae. sulcata, B. ovis ise R. bursa havuzlarında tespit edilmiştir.

Keywords

Theileria, Babesia, koyun, keçi, kene, RLB, Sivas

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