Diversity of two Theileria parva CD8+ antigens in cattle and buffalo-derived parasites in Tanzania

Theileria parva is a tick-transmitted protozoan parasite that causes a disease called East Coast fever (ECF) in cattle. This important tick borne-disease (TBD) causes significant economic losses in cattle in many sub-Saharan countries, including Tanzania. Cattle immunization using Muguga cocktail has been recommended as an effective method for controlling ECF in pastoral farming systems in Tanzania. However, immunity provided through immunization is partially strain-specific. Therefore, the control of ECF in Tanzania is still a challenge due to inadequate epidemiological information. This study was conducted to assess genetic diversity of Tp1 and Tp2 genes from T. parva isolates that are recognized by CD8 + T-cells in cattle and buffalo. The Tp1 and Tp2 genes are currently under evaluation as candidates for inclusion in a subunit vaccine. A total of 130 blood samples collected from cattle which do not interact with buffalo (98), cattle co-grazing with buffalo (19) and buffalo (13) in Mara, Mbeya, Morogoro, Tanga, and Coast regions in Tanzania were used in this study. Genomic DNA was extracted from the blood samples, Tp1 and Tp2 genes were amplified using nested PCR and the PCR products were purified and sequenced. The partial sequencing of the Tp1 and Tp2 genes from T. parva isolates exhibited polymorphisms in both loci, including the epitope-containing regions. Results for sequence analysis showed that the overall nucleotide polymorphism (π) was 0.7% and 13.5% for Tp1 and Tp2, respectively. The Tajima's D and Fu's Fs test showed a negative value for both Tp1 and Tp2 genes, indicating deviations from neutrality due to a recent population expansion. The study further revealed a low to high level of genetic differentiations between populations and high genetic variability within populations. The study also revealed that most samples from the seven populations possessed several epitopes in antigens that were identical to those in the T. parva Muguga reference stock, which is the main component of the widely used live vaccine cocktail. Therefore, different strategic planning and cost-effective control measures should be implemented in order to reduce losses caused by ECF in the study areas.