West Nile virus (WNV) is a mosquito-borne disease, usually present as a symptomatic disease but can cause various clinical signs ranged from mild fever to severe encephalitis and death in various animals and humans. In Egypt, the epidemiological data about WNV infection in different animal species particularly in domestic ruminants are scarce. The present study aimed to investigate the seroprevalence of WNV in cattle, buffalo, camel, sheep, and goats at some Governorates northern Egypt. In total, 360 serum samples (100 cattle, 50 buffalo, 50 camels, 85 sheep, and 75 goats) were examined using ELISA. The results revealed that the seroprevalence of WNV among ruminants was highly significant (P = 0.03) at Kafr El Sheikh Governorate (17.6%) in comparison with other the Governorates. Besides, the seroprevalence of WNV antibodies significantly differed between the examined species (P = 0.0001); it was 22%, 0%, 40%, 3.5%, and 5.3% in cattle, buffalo, camel, sheep, and goats, respectively. This is the first study to confirm that domestic ruminants act as a reservoir in the epidemiology of WNV infection and represent a risk for human and equine infections in Egypt