Family Rhabdoviride contains 2 genera Ephemerovirus and Lyssavirus which contain viruses responsible for two destructive diseases Bovine Ephemeral Fever (BEF) and rabies, respectively. Both diseases cause direct and indirect economic losses related to deaths, abortions, cost of treatment and prevention, zoonotic impact and restriction of animal movement. The main objective of this study is to evaluate the immune responses of cattle and buffalo vaccinated with BEF and rabies vaccines when administered separately in comparison with co vaccination. Serum samples were collected from 16 cattle and buffaloes (eight of each) then subjected to serum neutralization test SNT, serum biochemical, liver and kidney function for comparison according to the vaccinated groups. Each animal species divided into 4 groups (2 animals /group) group 1 was vaccinated by 2 doses of attenuated BEF vaccine inactivated at time of use with 2 weeks in between, group 2 was vaccinated by 1 dose of inactivated rabies vaccine, group 3 was vaccinated by rabies vaccine simultaneous with 1st dose of BEF vaccine and boostering dose of BEF vaccine after 2 weeks, group 4 not vaccinated and let to be control group. The results showed that both live attenuated BEF and inactivated cell culture rabies vaccines are safe because they didn't harmful effect on liver and kidney functions, they are immunogenic as they lead to significant increase in total serum protein due to increase in globulin. Co-vaccination of both vaccines together has higher levels of specific antibodies against both BEF and rabies in all vaccinated animals.