Bovine herpes virus type 1 (BoHV-1) is an economically important, worldwide distributed pathogen of domestic and wild Bovidae. It causes respiratory tract manifestations and/or abortion.The present study was achieved to study seroprevalence and make isolation of BoHV-1 from suspected cattle,buffaloes,sheep and goats collected from three governorates in Egypt (Menofya, Kalubeya and Dakahlia) through the years 2017- 2018.Serum and milk samples obtained from apparently healthy non-vaccinated animals were tested using indirect antibody detection ELISA. A total of 78 serum samples out of 346 samples and 46 milk samples out of 300 samples were positive. Virus was isolated from nasal swabs and lung tissue samples onchorioallantoic membrane (CAM) of 11-day-old embryonated chicken eggs. It showed congestion, edematous vacuoles and thickening small foci ranged from 2 to 3 mm in diameter, scattered on CAM membrane.Examination by electron microscope using positive staining methods showed the characteristic morphology of the herpes virion. The isolated virus was also genotypically confirmed to be BoHV-1 in a PCR assay using primers which specifically amplified a fragment (575 bp) of gC encoding gene in the viral DNA genome