Identification of Common Antigens in Babesia bovis, B. bigemina, and B. divergens

Abstract

Bovine babesiosis, caused by Babesia bovis, B. bigemina, and B. divergens, is a significant impediment to livestock production in countries with tropical/subtropical and temperate climates. Previous studies conducted on the immunoprophylaxis against the disease and diagnosis of these parasites has demonstrated the presence of similar antigens. The objective of this article was to identify and partially characterize antigens conserved among these three species. Immunochemical analysis using sera from cattle immunized individually with antigens from these three Babesia species revealed a number of antigens recognized by heterologous antisera. Cross-reactions were more evident in sera from cattle immunized with B. bovis/B. bigemina which recognized several antigens (15 kDa to >200 kDa) in B. divergens. Immunoscreening of a B. divergens cDNA library with bovine serum to B. bigemina allowed the isolation of five clones and DNA sequencing of plasmid BdJF5 showed a 680 bp cDNA insert. Basic Local Alignment Search Tool (BLAST) analysis of the predicted amino acid sequence revealed 47% identity with a protein identified as alphaNAC. Serum from mice immunized with a recombinant Glutathione S-Transferase-BdJF5 fusion protein immunoprecipitated a 20 kDa B. bovis antigen. However, 30 kDa and 18 kDa antigens were immunoprecipitated from B. divergens and immunoblotting analysis revealed the recognition of a 35 kDa B. bigemina antigen. An indirect fluorescence antibody assay on merozoites showed strong reaction with B. divergens and weak recognition of B. bovis and B. bigemina. Despite the existent antigenic polymorphism among the Babesia spp., these results demonstrated that common antigens occur between European B. divergens and Mexican B. bovis/B. bigemina.