Aycan Nuriye GAZYAĞCI, Ufuk KAYA, Sami GÖKPINAR, Meral AYDENİZÖZ

A research on Babesia and Theileria species in sheep and goats of Kırıkkale province through molecular methods

The aim of the present study was to determine presence and prevalence of Babesia and Theileria species in the sheep and goats of stock farming in the province of Kırıkkale, which is in Central Anatolia Region of Turkey through microscopic examination and RLB method. Venous blood samples of 3 mL were collected from vena jugularis of 300 sheep and 100 goats into tubes containing EDTA. Blood smears were prepared from samples; and stained by Giemsa and examined for Theileria and Babesia under a light microscope. Genomic DNAs were extracted from blood samples with EDTA through suitable methods; a region with a length of 360 to 430 bp on variable region V4 of the 18 ssu rRNA gene of Theileria and Babesia species were proliferated; PCR products obtained were hybridized on a membrane where species-specific probes. Piroplasms were detected in 54 (18%) sheep and in none of the goats during microscopic examination of blood smears. With the RLB technique; T. ovis 70%, Theileria sp. 2.3%, B. ovis 5.3%, T. ovis + B. ovis 2.7% were detected in sheep. In goats; T. ovis was detected at a rate of 1%. Consequently, this study is the first for detection of Theileria and Babesia species in sheep and goats in Kırıkkale. Presence of T. ovis and B. ovis in the sheep, and T. ovis in goats was detected in Kırıkkale.

PDF

___

1. Altay K, Ataş AD, Özkan E. Molecular survey of Theileria and Babesia species in small ruminants and ticks from Sivas region of Turkey. Manas Journal of Agriculture Veterinary and Life Sciences 2017; 7 (1): 30-39.
2. İnci A, Yıldırım A, Düzlü Ö. Kenelerin Medikal ve Veteriner Önemleri. Ed: İnci A, 1. Baskı. Kayseri, Türkiye: Erciyes Üniversitesi Yayınları; 2016.
3. Inci A, Yildirim A, Duzlu O, Doğanay M, Aksoy S. Tick-borne diseases in Turkey: A review based on one health perspective. PLOS Neglected Tropical Diseases 2016; 10 (12): e0005021.
4. Ahmed JS, Luo J, Schnittger L, Seitzer U, Jongejan F et al. Phylogenetic position of small ruminant infecting piroplasms. Annals New York Academy of Sciences 2006; 1081: 498–504.
5. Yin H, Schnittger L, Luo J, Seitzer U, Ahmed JS. Ovine theileriosis in China: a new look at an old story. Parasitology Research 2007; 101 (2): 191-195.
6. Ozubek S, Aktas M. Molecular and parasitological survey of ovine piroplasmosis, including the first report of Theileria annulata (Apicomplexa: Theileridae) in sheep and goats from Turkey. Journal of Medical Entomology 2017; 54 (1): 212–220.
7. Friedhoff, KT. Tick-borne disease of sheep and goats caused by Babesia, Theileria or Anaplasma spp. Parassitologia 1997; 39: 99-109.
8. Yin H, Liu Z, Guan G, Liu A, Ma M et al. Detection and differentiation of Theileria luwenshuni and T. uilenbergi infection in small ruminants by PCR. Transboundary and Emerging Diseases 2008; 55 (5-6): 233-237.
9. Alani AJ, Herbert IV. Pathogenesis of infection with Theileria recondita (Wales) isolated from Haemaphysalis punctata from north Wales. Veterinary Parasitology 1988; 28 (4): 293-301.
10. Uilenberg G. Babesia-a historical overview. Veterinary Parasitology 2006; 138 (1-2): 3-10.
11. Hashemi-Fesharki R. Tick-borne diseases of sheep and goats and their related vectors in Iran. Parassitologia 1997; 39: 115- 117.
12. Nagore D, Garcia-Sanmartin J, Garcia-Perez AL, Juste RA, Hurtado A. Identification, genetic diversity and prevalence of Theileria and Babesia species in a sheep population from Northern Spain. International Journal for Parasitology 2004; 34: 1059–1067.
13. Altay K, Dumanli N, Aktas M. Molecular identification, genetic diversity and distribution of Theileria and Babesia species infecting small ruminants. Veterinary Parasitology 2007b; 147: 161-165.
14. Altay K, Aktas M, Dumanlı N, Aydin MF. Evaluation of a PCR and comparison with RLB for detection and differentiation of Theileria sp. MK and other Theileria and Babesia species of small ruminants. Parasitology Research 2008; 103: 319–323.
15. Duh D, Punda-Polic V. Trilar T, Avsic-Zupanc T. Molecular detection of Theileria sp. in ticks and naturally infected sheep. Veterinary Parasitology 2008; 151: 327–331.
16. Aydin MF, Aktas M, Dumanli N. Molecular identification of Theileria and Babesia in sheep and goats in the Black Sea Region in Turkey. Parasitology Research 2013; 112: 2817–2824.
17. Tian Z, Liu G, Yin H, Xie J, Wang S et al. First report on the occurrence of Theileria sp. OT3 in China. Parasitology International 2014; 63: 403–407.
18. Giangaspero A, Marangi M, Papini R, Paoletti B, Wijnveld M et al. Theileria sp. OT3 and other tick-borne pathogens in sheep and ticks in Italy: molecular characterization and phylogeny. Ticks and Tick-Borne Diseases 2015; 6: 75–83.
19. Inci A, Ica A, Yildirim A, Duzlu O. Identification of Babesia and Theileria species in small ruminants in Central Anatolia (Turkey) via reverse line blotting. Turkish Journal of Veterinary and Animal Sciences 2010; 34 (2): 205-210.
20. Burridge MJ, Brown CG, Kimber CD. (1974) Theileria annulata: cross-reactions between a cell culture schizont antigen and antigens of East African species in the indirect fluorescent antibody test. Experimental Parasitology 1974; 35 (3):374–380.
21. Gubbels JM, De Vos AP, Van Der Weide M, Viseras J, Schouls LM et al. Simultaneous detection of bovine Theileria and Babesia species by Reverse line blot hybridization. Journal of Clinical Microbiology 1999; 37 (6):1782-1789.
22. Aktas M, Altay K, Dumanli N. Development of a polymerase chain reaction method for diagnosis of Babesia ovis infection in sheep and goats. Veterinary Parasitology 2005;133:277-281.
23. Gokpinar S, Aydenizoz M. Investigation of Theileria annulata and Theileria buffeli/orientalis in cattle from Kirikkale-Turkey by Reverse Line Blotting analyses. Israel Journal of Veterinary Medicine 2016; 71 (3): 20-25.
24. Georges K, Loria GR, Riili S, Greco A, Caracappa S et al. Detection of haemoparasites in cattle by reverse line blot hybridisation with a note on the distribution of ticks in Sicily. Veterinary Parasitology 2001; 31 (4): 273-286.
25. Gökpınar S. Kırıkkale yöresinde sığırlarda Theileria annulata ve Theileria buffeli/orientalis’in PCR-Reverse Line Blotting yöntemi ile araştırılması. Doktora Tezi. Kırıkkale Üniversitesi Sağlık Bilimleri Enstitüsü, Kırıkkale, Türkiye, 2013.
26. Schnittger L, Yin H, Qi B, Gubbels MJ, Beyer D et al. Simultaneous detection and differentiation of Theileria and Babesia parasites infecting small ruminants by reverse line blotting. Parasitology Research 2004; 92: 189–196.
27. Altay K, Aktas M, Dumanli N. Theileria infections in small ruminants in the east and southeast Anatolia. Acta Parasitologica Turcica 2007a; 31 (4): 268-271.
28. Bilgic HB, Bakırcı S, Kose O, Unlu AH, Hacılarlıoglu S et al. Prevalence of tick-borne haemoparasites in small ruminants in Turkey and diagnostic sensitivity of single-PCR and RLB. Parasites&Vectors 2017; 10: 211.
29. Karatepe B, Ozubek S, Karatepe M, Aktas M. Detection of Theileria and Babesia species in sheep and goats by microscopy and molecular methods in Nigde Province, Turkey. Revue de Médecine Vétérinaire 2019; 170: (7-9), 136-143.
30. Inci A, Nalbantoglu S, Cam Y, Atasever A, Karaer Z et al. Theileriosis and tick infestations in sheep and goats around Kayseri. Turkish Journal of Veterinary and Animal Sciences 2003; 27: 57-60.
31. Sayin F, Nalbantoglu S, Yukari BA, Cakmak A, Karaer Z. Epidemiological studies on sheep and goat Theileria infection. Veterinary Journal of Ankara University 2009; 56: 127-129.
32. Altay K, Dumanli N, Aktas M. A study on ovine tick-borne hemoprotozoan parasites (Theileria and Babesia) in the East Black Sea Region of Turkey. Parasitology Research 2012; 111:149–153.
33. Sevinç F, Dik B. The diagnosis of Babesia ovis in sheep in Konya around by ELISA. Eurasian Journal of Veterinary Sciences 1996; 12 (2): 73-79.
34. İnci A, Yukarı BA, Sayın F. Çankırı yöresinde bazı koyun ve keçi sürülerinde babesiosis ve theileriosis etkenlerinin mikroskobik kan muayenesiyle araştırılması. Veterinary Journal of Ankara University 1998;45: 105-113.
35. Aktas M, Altay K, Dumanli N. Survey of Theileria parasites of sheep in eastern Turkey using polymerase chain reaction. Small Ruminant Research 2005; 60: 289–293.
36. Sarayli H, Inci A, Ica A, Yildirim A, Duzlu O. Yeşilhişar yöresindeki koyun ve keçilerde Babesia etkenlerinin Reverse Line Blotting (RLB) yöntemiyle araştırılması. Erciyes Üniversitesi Sağlık Bilimleri Dergisi 2006; 15 (3): 181-188.
37. Aktas M, Altay K, Dumanli N. Determination of prevalence and risk factors for infection with Babesia ovis in small ruminants from Turkey by polymerase chain reaction. Parasitology Research 2007; 100: 797–802.
38. Shahzad W, Noor H, Ud-Din Ahmad M, Munir R, Sharif Shaghar M et al. Prevalence and molecular diagnosis of Babesia ovis and Theileria ovis in Lohi sheep at Livestock Experiment Station (LES), Bahadurnagar, Okara, Pakistan. Iranian Journal of Parasitology 2013; 8 (4): 570-578.
39. Yıldırım A, Düzlü Ö, İnci A, Önder Z. Çiloğlu A. Comparative diagnosis of Babesia bovis and Babesia bigemina in cattle by Reverse Line Blotting, Nested PCR and Real Time PCR techniques. Kafkas Universitesi Veteriner Fakultesi Dergisi 2013; 19 (5): 895-902.