Rabies, a Lyssavirus infection of Rhabdoviridae family, is a potential neurotropic disorder affecting all mammals and humans. This infection spreads through biting of infected and/or carrier animals to healthy ones including humans. Incubation period of this infection is quite variable ranging from a few days which can last up to one year in few cases. This case report presents the diagnosis and screening of suspected rabies samples of cow and mule. Clear behavioral changes along with paralysis of tail and hind legs were noticed in the mice of both groups between 11-15 days post inoculation while all the mice were found dead between 15-18 days post inoculation in mouse inoculation test (MIT). Reverse transcriptase-polymerase chain reaction (RT-PCR) performed for both samples gave a product of 443-bp amplifying the highly conserved “N”-region gene of virus confirming the rabies infection in both cases.