Brucellosis is one of the most common bacterial zoonotic diseases. There are two live attenuated vaccines used in control programs, B. abortus S19, B. melitensis rev1 vaccine for cattle,sheep and goat respectively . Some studies have observed one host infected with two different types of brucella. So in our study we try to prepare bivalent vaccine from two vaccinal strains to protect animal from virulent infection. Total of 150 brucella-free mice divided into six groupss , the first group was the control group , the second group was vaccinated with vaccine strain 19, the Third group was vaccinated with the vaccine rev1 , the fourth was injected with the bivalent vaccine without any adjuvant, the fifth was injected with the bivalent vaccine with mantonide Gel 1 (10%), and the sixth was injected with the bivalent vaccine with mantonide oil 206(1:1). The vaccinated mice were challenged with virulent strains after 30 day of vaccination, the 2nd group challenge B. abortus 544, the 3rd group challenge with B. melitensis 16M and the 4th, 5th and 6th were challenged the both strains. The immunogenicity of the vaccinated mice was measured after 15 day of challenge with virulent strains. It was found that the protective index of the 4th, 5th and 6th was (2.29, 2.53, 2.66 respectively) in mice when challenge with B. abortus 544, and was (2.41, 2.53, 2.68 respectively) in mice when challenge with B. melitensis 16M. The prepared vaccines were potent in comparison with PBS inoculated (unvaccinated) group.