Identification of different Theileria species (Theileria lestoquardi, Theileria ovis, and Theileria annulata) in naturally infected sheep using nested PCR–RFLP

Abstract

Ovine theileriosis is an important hemoprotozoal disease of sheep and goats in tropical and subtropical regions that leads to economic losses in these animals. A nested PCR-restriction fragment length polymorphism (RFLP) was carried out to identification Theileria species in sheep in some area in western half of Iran (Sari, Rasht, Urmia, Ilam, and Ahvaz). Two hundred and fifty blood samples were taken from sheep during tick activating season (summer of 2008). Microscopic examination revealed that 9.2% (23/250) sheep were infected by Theileria spp. piroplasms. Parasitemia ranged from 0.011% to 0.015%. In nested PCR assessment of DNA samples, 32.8% (82/250) sheep were positive. The negative samples were confirmed by amplifying of ovine beta-actin gene as an internal control. The differentiation of Theileria species was based on RFLP patterns using three restriction enzymes: HpaII, Rsa1, and Bsh 1285I. Out of 82 positive samples, 54.8% (45/82) and 40.2% (33/82) were positive for Theileria lestoquardi and Theileria ovis respectively. Mixed infection was detected in 4.8% (4/82) cases. Based on their PCR product digestion pattern with HpaII (1178, 900, 278, and 106 bp), it seemed to be mixture of Theileria annulata and T. lestoquardi. The presence of T. annulata was supported by sequence analysis. This is the first report of naturally infected sheep with T. annulata in Iran. Geographical distribution of Theileria species in sheep is shown according to the result of microscopy and nested PCR and RFLP data.

A study on prevalence and identification of Ovine Theileria and Babesia infection in Zabol using PCR method

“…In addition Low prevalence of Babesia in other countries such as Turkey and Germany have also been reported Theodoropoulos et al 2006). Zaeemi et al (2011) andHeidarpour-Bami et al (2010) in a study on the prevalence of Theileria species by PCR-RFLP method in west and east of Iran determined the frequencies of Theileria spp. infection in sheep in the range of 32.8-60 %, respectively.…”

Detection of Theileria annulata in blood samples of native cattle by PCR and smear method in Southeast of Iran

“…Recently, diagnostic methods like PCR have been developed for the rapid and accurate detection of Theileria spp. (Zaeemi et al 2011, Ghaemi et al 2012.…”

Molecular surveillance of Theileria parasites of livestock in Oman

“…(Fig 2). Aydin et al, 2015Aydin et al, , 2013 (Zaeemi et al, 2011;Jalali et al, 2014), in the Sudan, where 7.8% of infected sheep 310 carried mixed infection of T. annulata and T. lestoquardi (Taha et al, 2013) …”