Prevalence, First Molecular Identification and Characterization of Theileria lestoquardi in Sheep in Alhuda National Sheep Research Station, Al Gezira State, Sudan

Abstract

Aims: The present study investigated the presence of T. lestoquardi in Alhuda National Sheep Research Station using three diagnostic techniques, identified and characterized the parasite. Methodology: Forty-five animals from three Sudanese sheep ecotypes were selected randomly during September 2013. Blood and sera samples were examined by microscopy, indirect fluorescent antibody test (IFAT) and polymerase chain reaction (PCR). The PCR product was sequenced to confirm the identity of the parasite. Results: Microscopic examination (ME) revealed the presence of Theileria spp. piroplasms in 31 animals out of 45 (68.9%).Sixteen sera samples out of the 45 were tested for antibodies against T. lestoquardi, fourteen of these (87.5%) were found positive by IFAT. PCR targeting Theileria lestoquardi merozoite/piroplasm surface antigen (Tlms) was positive in 4/45 (8.9%). Prevalence of the disease was significantly higher (P<.05) in females and in >1 year age group by ME. Sequence analysis confirmed the species identity when compared with reference sequences retrieved from GenBank Database and indicated 99% homology. Multiple sequence alignment of Alhuda strain with other T. lestoquardi sequences obtained from Sudan and Iran revealed five silent base pair substitutions. Phylogenetic tree showed clustering of Alhuda strain (accession no. KY965145) with other T. lestoquardi sequences in one distinct clade. Conclusion: To the best of our knowledge this is the first report on molecular identification and characterization of T. lestoquardi from Al Gezira State, Sudan. The disease could adversely affect the animal population in the state therefore epidemiological surveys are required to build up data necessary for designing efficient control programs.