Sequel to the 2024 highly pathogenic avian influenza (HPAI) virus H5N1 outbreak among cattle in the United States of America, it became imperative to investigate the exposure of Nigerian cattle, a major source of much needed animal protein, to avian influenza virus (AIV). This study adopted a cross-sectional design to evaluate influenza A virus (IAV) exposure among the tested cattle using serological and molecular techniques. We collected 693 blood and 54 milk samples from six states located in the south-western, north-central and north-eastern 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 the cattle seropositivity was not due to the 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.