Five antigen preparations from Brucella ovis strain REO 198 were incorporated with the pluronic polymer L-121 and muramyl dipeptide and tested as vaccines against B. ovis infection of rams. The antigenic preparations were: (1) a fraction enriched in outer membrane proteins and rough lipopolysaccharide (hot saline extract, HS); (2) the proteins from HS substantially free of lipopolysaccharide; (3) outer membrane blebs; (4) outer membrane-peptidoglycan complexes extracted with detergent; (5) killed whole cells. The experimental vaccines were compared with two standard vaccines, rough Brucella abortus 45/20 whole killed cells in an oil based adjuvant, and live Brucella melitensis Rev 1. Immunizations with non-living vaccines were performed on two occasions, 18 weeks apart. The rams were challenged with a virulent strain of B. ovis 31 weeks after the second vaccination and slaughtered 15 weeks thereafter. Rates of infection in groups vaccinated with Rev 1 (33%), and HS (40%) were significantly lower (P < 0.005 and P < 0.025, respectively) than that in the non-vaccinated control group (87%). Strain 45/20 was the only other vaccine that conferred a significant level of protection (50%) (P < 0.05). The organ distribution of the infection and the level of colonization of infected organs did not differ significantly between infected animals in the various vaccine groups and those in the unvaccinated control group. No statistically significant relationship was detected between the magnitude of the antibody responses to the HS extract, to outer membrane proteins, or to the rough lipopolysaccharide, and freedom from infection. The results indicate that the HS extract of B. ovis may represent a useful alternative to B. melitensis Rev 1 or B. abortus 45/20 as a vaccine against B. ovis.