Molecular identification and antigenic characterization of Babesia divergens Erythrocyte Binding Protein (BdEBP) as a potential vaccine candidate

Host cell invasion is the only step where Babesia parasites are extracellular, and their survival is menaced during this step. Therefore, interfering with this critical stage is a target for an anti-Babesia intervention strategy. In this regard, recombinant protein encoding Babesia divergens Erythrocyte Binding Protein (BdEBP) was produced in Escherichia coli in the current study, and its antiserum was prepared in mice for further molecular characterization. Western blotting and indirect fluorescent antibody test (IFAT) revealed the specific reaction of the anti-rBdEBP serum with a corresponding authentic protein of B. divergens. Next, bovine RBCs were incubated with a B. divergens lysate, and anti-rBdEBP serum was produced in mice to detect the ability of BdEBP to bind with host cells. Bands corresponding to 29.6-kDa proteins in the protein-bound erythrocyte lysate were detected by specific immune rBdEBP using Western blotting. These results suggest that BdEBP is functional in the merozoite stage and may be involved in attachment to bovine RBCs. A significant inhibition of the in vitro growth of B. divergens culture treated with anti-rBdEBP serum was observed. Moreover, the efficacy of pre-incubated free merozoites to invade bovine erythrocytes was inhibited by 60% after incubation with 2mg/ml of anti-rBdEBP serum for 6h. The obtained data suggest the possible use of rBdEBP as a vaccine candidate against bovine babesiosis.