The production of monoclonal antibodies to the bloodstages of the haemoprotozoan parasites Babesia caballi and Babesia equi and the characterization of their corresponding antigens are described. Species specific and immunogenic proteins of both parasites were identified using SDS-PAGE, Western blotting and ELISA. These proteins were then electroeluted from SDS-PAGE gels and used to immunize BALB/c mice for hybridoma production. One monoclonal antibody (Mab), designated BC22.214.171.124 (BC5), recognized a 70 kDa protein of B. caballi as demonstrated by Western blotting under reducing conditions. Another Mab, BE1.24/2.95 (BEI), recognized a 34 kDa protein of B. equi. Both Mabs reacted specifically in indirect ELISA when isolated whole merozoites were used as antigen. Preliminary studies using the two Mabs in a competitive ELISA (cELISA) suggest that the cELISA for the detection of B. caballi infection is more sensitive than the commonly used complement fixation test but that refinement is necessary for the B. equi system.