PCR-RELP for detecting of Theileria annulata infection in cattle and Hyalomma species in Kermanshah Province, Iran

Abstract

Bovine theileriosis is important disease in tropical and subtropical areas with great economic losses in livestock husbandry in Iran. The aim of study was to assess the prevalence of Theileria annulata infection in cattle and Hyalomma species of Kermanshah Province, Iran. A number of 138 blood samples were randomly taken from examined cattle. The genomic DNA was extracted and PCR was performed to specifically amplify a 721-bp-long fragment of the 30 Kilo Dalton major merozoite surface antigen (30 KDa msa) of T. annulata. The amplified products were digested with TaqI, Rasl, and AluI restriction enzymes. Overall prevalence was 9.44% (13/138) with lymphadnopathy (1.17%) and pale mucosal membrane (1.9%) in Holstein cattle aged <1 year and more than 5 years-old, respectively. Five species of genus Hyalomma (18.8%, 141/750) including Hyalomma anatolicum anatolicum (30.4%), H. anatolicum excavatum (31.2%), H. marginatum (36.8%), H. asiaticum asiaticum (0.7%), and H. detritum (0.7%) were identified. The tick indices for each Hyalomma species were ranged from 0.01 to 0.36. PCR findings indicated that 3 out of 138 blood samples (2.17%) and 19 out of 141 Hyalomma ticks (4.13%) were infected with T. annulata. Amplified PCR products from blood samples generated similar RFLP patterns, but different RFLP pattern for T. annulata from H. anatolicum anatolicum (9.21%) and H. anatolicum excavatum (4.2%). The RFLP patterns of the amplified fragment of the 30 KDa msa of T. annulata indicated the circulation of four different T. annulata isolates of H. anatolicum anatolicum and H. anatolicum excavatum in the region.