Articles

Molecular Detection and Identification of Theileria Species by PCR-RFLP Method in Sheep from Ahvaz, Southern Iran

Abstract

 

Background: The present study was carried out to investigate the accurate status of ovine Theileria infection in sheep from Ahvaz and surrounding region, a tropical area southwest Iran.

Methods:A PCR-RFLP method based on 18S ribosomal RNA gene was designed which could detect and differentiate Theileria and Babesia spp. and also differentiate main Theileria species in sheep at the same time. 119 sheep blood samples were col-lected from Ahvaz and surroundings.

Results: Microscopic examination of blood smears revealed 69.7% (83/119) infec-tion with Theileria spp. Of the total samples subjected to PCR, 89% (106/119) were found to be positive, all of which were identified as Theileria by RFLP analysis using enzyme Hind II. In enzymatic digestion of PCR products by Vsp I, 91.5% (97/106) of Theileria positive samples were identified as T. ovis while mixed Theileria infections were found in 9 samples. The samples with mixed infections were analyzed with an additional nested PCR-RFLP method, by HpaII enzyme digestion. 3 samples with T. lestoquardi infection, 1 sample with T. ovis and T. annulata, 1 sample with T. lestoquardi and T. annulata, and 4 samples with T. ovis, T. lestoquardi and T. annulata mixed infections were detected.

Conclusion: Ovine theileriosis caused by T. ovis is highly prevalent in southwest Iran while T. lestoquardi and T. annulata infection can be detected in a lesser propor-tion of sheep in this region. The new PCR-RFLP method that was designed in this study, can serve as a beneficial diagnostic tool, especially in T. ovis prevalent re-gions.

Aktas M, Altay K, Dumanli N. Survey of Theileria parasites of sheep in eastern Turkey using polymerase chain reaction. Small Rum Res. 2005; 60: 289–293.

Gao YL, Yin H, Luo JX, Ouyang WQ, Bao HM, Guan GQ, Zhang QC, Lu WS, Ma ML. Development of an enzyme-linked im-munosorbent assay for the diagnosis of Theileria sp. infection in sheep. Parasitol Res.2002; 88: 8-10.

Altay K, Aktas M, Dumanli N. Theileria in-fections in small ruminants in the East and Southeast Anatolia. Türkiye Parazitoloji Dergisi.2007; 31: 268–271.

Hashemi-Fesharaki R. Tick-borne diseases of sheep and goats and their related vectors in Iran. Parasitologia.1997; 39: 115-117.

Hooshmand-Rad P, Hawa NJ. Malignant theileriosis of sheep and goats. Trop Anim Health Prod.1973; 5: 97-102.

HeidarpourBami M, Khazraiinia P, Had-dadzadeh HR, Kazemi B. Identification of Theileria species in sheep in the eastern half of Iran using nested PCR-RFLP and mi-croscopic techniques. Iranian J Vet Res. 2010; 11: 262-266.

Razmi GR, Hossieni M, Aslani MR. Identi-fication of tick vectors of ovine theileriosis in an endemic region of Iran. Vet Parasitol Res.2003; 116: 1-6.

Spitalska E, Namavari MM, Hosseini MH, Shad-del F, Amrabadi OR, Sparagano OAE. Molecular surveillance of tick-borne diseases in Iranian small ruminants. Small Rum Res.2005; 57: 245-248.

Zaeemi M, Haddadzadeh H, Khazraiinia P, Kazemi B, Bandehpour M. Identification of different Theileria species (Theileria lestoquardi, Theileria ovis, and Theileria annulata) in natu-rally infected sheep using nested PCR–RFLP. Parasitol Res.2011; 108: 837–843.

Haddadzadeh HR, Rahbari S, Khazraiinia P, Nabian S. New concepts on limiting factors of ovine and caprine malignant theileriosis (OCMT) in Iran. Iranian J Vet Res.2004; 5: 43-46.

Inci A, Ica A, Yildirim A, Duzlu O. Identi-fication of Babesia and Theileria species in small ruminants in Central Anatolia (Tur-key) via reverse line blotting. Turk J Vet. Anim Sci. 2010; 34: 205-210.

Altay K, Dumanlia N, Holmanb PJ, Aktas M. Detection of Theileria ovis in naturally in-fected sheep by nested PCR. Vet Parasi-tol.2005; 127: 99–104.

Shayan P, Rahbari S. Simultaneous differen-tiation between Theileria spp. and Babesia spp. on stained blood smear using PCR. Parasitol Res.2005; 97: 281–286.

Kirvar E, Ilhan T, Katzer F, Wilkie G, Hooshmand-Rad P, Brown D. Detection of Theileria lestoquardi (hirci) in ticks, sheep, and goats using the polymerase chain reaction. Ann NY Acad Sci.1998; 849: 52–62.

Gubbels MJ, Hong Y, Van Der Weide M, Qi B, Nijman IJ, Guangyuan L, Jongejan F. Molecular characterisation of the Theileria buffeli/orientalis group. Int J Parasitol.2002; 33: 943–952.

Al-Amery MAY, Hasso SA. Laboratory diagnosis of novel species of Theileria hirci, Eimeria caprovina and Eimeria pallida in goats in Iraq. Small Rum Res.2002; 44: 163–166.

HeidarpourBami M, Haddadzadeh HR, Kazemi B, Khazraiinia P, Bandehpour M, Aktas M. Molecular identification of ovine Theileria species by a new PCR–RFLP method. Vet Parasitol.2009; 161: 171–177.

Razmi GR, Eshrati H, Rashtibaf M. Preva-lence of Theileria spp. infection in sheep in South Khorasan province, Iran. Vet Parasi-tol.2006; 140: 239–243.

Al-Alousi TL, Hayatee ZG, Latif BM.A. Incidence of Theileriasis in sheep of Mosul area in Iraq. J Vet Parasitol.1988; 2: 149–150.

Durrani AZ, Younus M, Kamal N, Mehmood N, Shakoori AR. Prevalence of Ovine Theileria Species in District Lahore, Pakistan. Pakistan J Zool.2011; 43: 57-60.

Salih DA, El Hussein AM, Hayat M, Taha KM. Survey of Theileria lestoquardi antibodies among Sudanese sheep. Vet Parasitol.2003; 111: 361-367.

Leemans I, Brown D, Hooshmand-Rad P, Kirvar E, Uggla A. Infectivity and cross-im-munity studies of Theileria lestoquardi and Theileria annulata in sheep and cattle: I. In vivo responses. Vet Parasitol.1999; 82: 179-192.

Files
IssueVol 9 No 1 (2014) QRcode
SectionArticles
Keywords
18S rRNA gene Iran PCR-RFLP Sheep Theileria spp.Babesia

Rights and permissions
Creative Commons License This work is licensed under a Creative Commons Attribution-NonCommercial 4.0 International License.
How to Cite
1.
Jalali SM, Khaki Z, Kazemi B, Rahbari S, Shayan P, Bandehpour M, Yasini SP. Molecular Detection and Identification of Theileria Species by PCR-RFLP Method in Sheep from Ahvaz, Southern Iran. Iran J Parasitol. 1;9(1):99-106.