Molecular Identification and Genotyping of Theileria orientalis type 3 (buffeli) from cattle in Guilan Province of Iran, using Nested-PCR assay

Document Type: Original Articles


1 Department of Parasite Vaccine Research and Production of Razi Vaccine and Serum Research Institute, Agriculture Research, Education and Extension Organization (AREEO), Karaj, Iran

2 Veterinarian in private clinical practice, Takistan, Qazvin Province, Iran


Protozoan parasites of the genus Theileria are tick-borne parasites that have been found in many species of mammals. More than a dozen species of Theileria occur in cattle, water buffalo, sheep and goats. Theileria orientalis is a non-pathogen blood protozoan parasite, was detected and identified during a regular investigation of piroplasmida infection in indigenous cattle in spring season of 2019 from Northern Provinces of Iran. A total of 92 blood samples were collected in the spring season from different areas of Guilan and Mazandaran Provinces, Iran. The Giemsa stained blood smears did not show any parasite infection but T. orientalis was identified by 18S rRNA gene PCR and DNA sequencing. The specific sequenced DNA for T. orientalis was registered in GenBank under accession number of MN453385. The partial 18S rRNA gene sequence of obtained DNA has showed 100% nucleotide identity with reference sequences for the T. orientalis have been registered from Europe, Africa and Asia. Additionally, molecular phylogenetic studies have shown that T. orientalis Iran GC98-01 isolate belongs to nonpathogenic T. orientalis type 3 (buffeli). In this study, the indigenous Bos indicus cattle were detected as asymptomatic carrier state for Theileria spp infection. Here we identified and genotyped T. orientalis for the first time as T. orientalis type 3 (buffeli) from Iran using molecular phylogenetic analysis and the 18S rRNA gene sequence of the T. orientalis GC98-01 isolate was registered in GenBank. Moreover, rare T. annulata infection was detected in cattle using semi nested-PCR in Mazandaran (Miankaleh peninsula). T. orientalis can be differentiated from other Theileria and Babesia haemoprorotozoan parasites by specific molecular assay.