Equine hepesirus-1 (EHV-1) is among the infectious diseases that threaten equine health. EHV-1 infects horses causing epidemic respiratory disease, abortion in pregnant mares, neonatal foal death, myeloencephalopathy, and chorioretinpathy. The present study was designed to investigate the presence of EHV-1 in blood serum of horses and donkeys in different localities in Qalubiah Governorate. A total of 185 blood serum samples were collected from horses and donkeys. All animals were evaluated based off whether they were of Indigenous breeds, different ages, apparent health, and clinical signs to equine herpes. The collected blood serum samples were investigated against equine herpes virus type-1 by using an indirect ELISA. The results revealed that 39.5% (73/185) of all horses and donkeys’ sera samples were EHV-1 seropositive while 42.86 % (33/77) of the horses’sera samples and 37 % (40/108) of the collected donkeys’sera samples were EHV-1 seropositive. Meanwhile, 57.14 % (44/77) in horses and 63 % (68/108) in donkeys’ sera samples were found to be seronegative for the EHV-1 infections. Analysis of data revealed that, young animals (