Influenza A is an emerging zoonotic virus with worldwide distribution. To our knowledge, no studies have been conducted to assess influenza A exposure in stray cats in regions with positive cases of wild birds. This study aimed to determine the seroprevalence of anti-influenza A antibodies in feral cats from a region in Spain with cases of positive wild birds. A cross-sectional study of stray cats (n = 183) was conducted between March 2022 and March 2023. The presence of antibodies against the influenza A virus was tested using a commercial enzyme-linked immunosorbent assay kit adapted for this study and confirmed by competitive enzyme-linked immunosorbent assay for the detection of antibodies against the haemagglutinin H5. During sample collection, none of the cats exhibited clinical signs of illness. Four of the 183 animals tested showed anti-influenza A antibodies by ELISA, and the seroprevalence of influenza A was 2.19% (95% confidence interval 0.85%-5.48%). Due to the low number of positive cases detected, it appears that cats did not have an important epidemiological role in influenza A transmission during this period.