Bovine ephemeral fever virus (BEFV),an economically important arthropod-borne viral pathogen,causes bovine ephemeral fever (BEF) in cattle and water buffalo. In the current study, we detected the BEFV in threeEgyptian governorates(Giza, Menofia and Sharqiyah) throughisolation, serological and molecular identification of the virus from naturally infected cattle. Fifty-five cattle showed symptoms suspected to be BEF represented by fever, lameness, subcutaneous emphysema, and recumbence. Serum samples were obtained for evaluation the immune status of affected cattle and buffy coat samples were subjected to trials of virus isolation on baby hamster kidney (BHK-21) cell line and also inbaby mice where positive results were recorded among 10 samples only (4 samples form Giza, 3 samples from each Menofia and Sharqiyah).SNT results revealed that all the infected cattle had very low titers of BEFV serum neutralizing antibody which indicated that such were fully susceptible to the infection.Serological identification including direct fluorescent antibody technique (dFAT) and virus neutralization assay (VN)confirmed that the obtained viral isolatesare BEFV. Further confirmation by reverse trascriptase polymerase chain reaction (RT-PCR) targeting the glycoprotein (G) gene showed positive amplification reaction of the target geneat the correct size of 500bp. Consequently, further future work for molecular analysis of such recent isolates will be conducted for evaluation of the circulating BEFV in these governorates in Egypt.