This study was conducted to evaluate the protective effect of Brucella melitensis Rev.1 and the use of BCG vaccine as immunostimulant by subcutaneous injection in Guinea pigs. Lab. animals were divided into 8 groups: Combined Rev.1 and BCG vaccines injected in the1 st group & both vaccines were injected simultaneously in the 2 nd group. Three groups sensitized with BCG vaccine then injected with Rev.1 vaccine one week, two week & three weeks intervals respectively. Other two groups were injected with BCG and Rev.1 vaccine individually. The last one was unvaccinated control. All injected animals showed resistance to infection with 16 M strain (90 %, 80 %, 80, 60 %, 70 %, 0 % and 70 % respectively).Thus animals vaccinated with bivalent Rev.1 and BCG vaccines (in one shot) showed the best protective level to infection.

We evaluated orally administered live attenuated B. melitensis Rev.1 vaccine in combination with flagellin to protect mice against I/P challenge with Brucella melitensis 16M bacteria. Optimal protection was enhanced by three booster immunization doses against challenge at 3 weeks post challenge. Experiments were performed with mice to elucidate the roles of humoral and cell-mediated immune responses in the acquisition of protective immunity to Brucella melitensis and to compare infection immunity with immunity developed through vaccination with oral reduced dose of Rev.1 vaccine of B. melitensis combined with flagellar protein (H7). Vaccination with reduced dose of Rev.1 vaccine orally combined with flagellar protein is better than the vaccination with full dose of Rev.1 vaccine S/C in mice. From the results its pointed out that the cell mediated immunity elicited by the use of oral reduced dose of Rev.1 vaccine (3 Successive) dose combined with flagellar protein.

In this work five vaccination protocols were prepared and applied these protocols in sheep. Serum samples were collected from each group at time points 25, 50, 77, 89, and 98 days post vaccination. The results revealed that humoral antibodies were detected in groups 3, 4 and 5 by using Rose Bengal test (RBT), Buffer Acidified Plate Antigen Test (BAPAT), and Tube Agglutination Test at 98 days, 77 days, and 77 days respectively, while for ELISA test revealed that were positive at 50, 89, 98, 98, and 98 days post vaccination in group 1, 2,3,4 and 5 respectively. Cell mediated immunity was evaluated by Lymphocyte Blastogenesis Assay Test and Brucellin test (Delayed Type Hyper Sensitivity Test). The results indicated that there were no significant differences in between mean of different groups at P ≤ 0.05, So for Skin Delayed Hyper Sensitivity test, Group 1 and group 6 were negative while Group 2, 3, 4 and 5 were positive. Conclusion, animals in Groups 3, 4 and 5 had humoral immune response and can be protected from abortion in pregnant ewes and prevent infection. In this work, we evaluated to potential of three doses reduced Rev.1 mixed with E. coli flagellin which induced protection without need of adjuvant against I/P Brucella melitensis challenge. Also these data suggest that flagellin proteins might induce protective immune responses and these proteins will be a good candidate for subunit vaccine against ovine brucellosis in sheep.