Antigen profiles of Anaplasma ovis and A. mesaeterum were analysed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and immunoblotting, and cross infection trials were performed by A. ovis challenge to A. mesaeterum- or A. marginale-inoculated goats and A. marginale challenge to an A. ovis-inoculated calf. Antigen analysis showed unique and common proteins of A. ovis and A. mesaeterum and identified a major 38 kDa protein to be a dominant immunogen bearing two epitopes common to Anaplasma species. An epitope specific to A. ovis and A. marginale, absent from A. mesaeterum, was recognised on the major protein of A. ovis. Pre-inoculation of goats with A. mesaeterum induced continuous antibody response with low parasitemia and partial protection against A. ovis challenge, as demonstrated by lower peak parasitemia and normal body temperature compared with values for goats inoculated with A. ovis alone. Pre-inoculation of goats with A. marginale and a calf with A. ovis induced weak or no antibody response without parasitemia, and was of no effect for preventing anaplasmosis by heterologous challenge. These results suggest that the development of parasitemia is essential for inducing continuous antibody by which cross protection would be possible.