Equine influenza virus (EIA) type A virus (H3N8 and H7N7) is a leading cause of serious respiratory disease in horses causing worldwide morbidity and economic losses. In this study, serological evidence of EIV subtype H3N8 was investigated in horses and donkeys in 4 Egyptian provinces (Cairo, Giza, Sharkia, and Gharbia) between (December 2017 and January 2019). Serum samples of 164 horses and 98 donkeys were examined by ELISA and HI tests giving 63 positive horse serum samples (38.41%) for EIV antibodies, while 32 donkey serum samples were positive (32.65%). Sharkia province showed the highest EI prevalence in horse serum samples with a 47.87 % prevalence rate followed by Gharbia, Cairo, and Giza provinces with 40%, 11.11%, and 11.76% respectively by ELISA. Furthermore, Sharkia province also showed the highest EI prevalence rate (44.44 %) in donkey sera followed by Gharbia and Cairo provinces with (23.07%), and (16.67 %) respectively by ELISA. The results of this study showed a higher predominance of the virus antibodies in horses in the winter season at 43.18% and a lower rate in summer with 27.94% by ELISA. Considering the age, EIV antibodies varied considerably in different age groups by ELISA, animals 1-3 years old showed the highest EIV prevalence (43.96) %. while the lowest rate (24.14 %) was observed in animals younger than one year. in contrast, HI test results showed variations in different age groups; animals less than one-year-old showed the highest viral prevalence, with an average of (8.62 %). While the lowest prevalence rate (1.13 %) was observed in animals older than three years.