Epizootic Hemorrhagic Disease Virus (EHDV) impacts deer and livestock operations globally. In this study, we tested mono-, bi-, and tri-valent vaccines for three serotypes of EHDV in farmed white-tailed deer (Odocoileus virginianus) in Florida. We compared serological status and antibody titers from 39 previously unvaccinated white-tailed deer at three time points. Seven animals were 14 weeks old and 32 individuals were 15 months of age. At day 0, we injected the first dose and collected blood samples; at day 14, we injected a booster and collected blood samples; and at day 33, we collected a final blood sample. We found low titer levels on days 0 and 14, with some outliers. By day 33, most animals in the treatment group responded with a high titer of homologous antibody. Animals injected with a placebo did not show a similar response. At day 33, the difference between treatment and placebo group titers and number of animals that were seropositive were significantly different. We found no difference in titer levels for vaccines when they were administered as mono-, bi- or tri-valent forms. During the course of this study 4 animals died of natural EHDV infection, but none of them received a full vaccine dose. Homologous antibody responses of deer and opportunistic challenge with naturally occurring EHDV suggest that this vaccine may stimulate the immune system sufficiently to protect individuals.