Following the avian influenza (AI) epidemics occurring in different areas of the world, a surveillance program funded by the Italian Ministry of Health was implemented. In the framework of this program, an investigation of wild birds was carried out to assess the circulation of AI viruses in their natural reservoir. More than 3000 samples, mainly cloacal swabs, were collected from migratory wild birds belonging to the orders Anseriformes and Charadriiformes. Samples were screened by means of a real-time reverse transcriptase polymerase chain reaction (RRT-PCR), then processed for attempted virus isolation in embryonated fowl's specific pathogen-free eggs. Approximately 5% of the samples were positive for type A influenza viruses by RRT-PCR, and from 14 of those samples AI viruses were isolated and fully characterized. The isolates, belonging to 8 different avian influenza virus subtype combinations (H10N4, H1N1, H4N6, H7N7, H7N4, H5N1, H5N2, and H5N3), were obtained from migratory Anseriformes.