Protective vaccination against virulent Babesia bovis with a low-molecular-weight antigen

A Babesia bovis low-molecular-weight antigen was purified from crude material by using affinity adsorption techniques first with a mouse monoclonal antibody and then by adsorption with normal bovine sera. The antigen was then further purified by gradient gel electrophoresis. Analysis by Western transfer revealed only one antigen band, with an apparent molecular mass of 29 kilodaltons. A band (12 by 0.3 by 0.6 cm) corresponding to this antigen was excised from the acrylamide gel and injected twice 4 weeks apart, together with 2.5 ml of Freund complete adjuvant, into nine nonsplenectomized adult cattle. The vaccinated cattle and five susceptible control animals were challenged with a virulent homologous strain 4 weeks after the second vaccination. None of the vaccinated animals was clinically affected, whereas three of five controls were severely affected. Control animals had significantly greater declines in packed cell volume and greater rises in temperature and parasitaemias than vaccinated animals.