This study was designed to investigate the exposure of Nigerian cattle to influenza A virus (IAV) using serological and molecular techniques, considering the 2024 detection of Highly Pathogenic Avian Influenza (HPAI) H5N1 among cattle in the United States of America. We collected 693 blood and 54 milk samples from six Nigerian states spread across the southwest, north-central and north-east regions of Nigeria. Of these, 31 blood samples (4.5%) were seropositive for IAV nucleoprotein, whereas all milk samples tested negative. Moreover, the IAV matrix gene was not detected in any of the samples. Our findings showed that cattle seropositivity was not due to HPAI H5 exposure. To our knowledge, this represents the first evidence of IAV exposure among cattle in Nigeria. Further research is warranted to identify the risk factors that facilitate intra- and interspecies transmission of IAV within the Nigerian cattle population.