In April 2009, a new influenza A virus (IAV) subtype (A(H1N1)pdm09) spread worldwide and triggered the first human influenza pandemic of the 21st century. Since then, exposure to the pandemic H1N1 IAV has been confirmed in different animal species. Serological evidence and clinical infection with A(H1N1)pdm09 have been reported in canines, but the information available about the role of dogs in the epidemiology of this IAV subtype is still very limited in Europe. A cross-sectional study was carried out to determine the seroprevalence of A(H1N1)pdm09 in dogs in southern Spain, a region with endemic seasonal circulation in human. Sera from 750 companion dogs were collected during the period 2013-2016. Antibodies against pandemic H1N1 IAV were analysed using the haemagglutination inhibition test. Positive samples were also tested by single radial haemolysis assay. Seropositivity was only confirmed by both methods in one (0.13%; 95% CI: 0.00-0.38) adult animal sampled in 2013. To the best of the authors´ knowledge, this is the first report of A(H1N1)pdm09 exposure in dogs in Spain. The low seroprevalence obtained indicates a limited exposure history to A(H1N1)pdm09 IAV in dogs in this country and suggests a low risk of transmission of this zoonotic IAV subtype between humans and dogs.