Bovine ephemeral fever (BEF) is an infectious, arthropod-born viral disease has economic importance in cattle and buffaloes. Vaccination is widely used to control and prevent BEF disease. This study aimed to prepare an inactivated BEF vaccine using Montanide ISA 206 (water-in-oil-in-water) adjuvant that was applied in two different protocols (one dose and two doses one month apart). The prepared vaccine was sterile and safe. It was found that a single dose of the vaccine induced protective neutralizing serum antibody titer from 2nd week post vaccination (PV), reached highest titer at 10th week PV and persisted in this protective titer until 34 weeks PV using SNT and confirmed using ELISA, however boostering of animals 4 weeks post preliminary vaccination increased the titer of the protective neutralizing antibodies and its time duration to 44 weeks PV. Thus, in order to provide immunity that will last the entire season it is recommended that the vaccine should be administered short time before the onset of BEF season (summer season).