A total of 143 serum samples were collected from migratory birds captured or hunted during Sep. 2012 – Mar. 2013 at 4 distant sites located in Aswan to investigate the presence of avian influenza virus (AIV) antibodies. The migratory birds were classified into (8) orders, (16) families and (23) species. The serum samples were examined using IDEXX MultiS-screen AI Antibody ELISA kit coated with AIV nucleoprotein and hemagglutination inhibition (HI) test against H5 and H9 antigens. The overall results using both ELISA and HI tests revealed that the prevalence of AIV antibodies was 12.6% (18/143). The seropositive samples represented 5/143 and 15/108 for ELISA and HI, respectively. The detection of both H5 and H9 antibodies in (4) samples indicated that co-infection may occur, while (10) samples were only containing H9 antibodies and (1) sample was H5 positive by HI test. The AIV positive samples were detected in (9) species belongs to (5) orders and (7) families. The Anseriformes and Charadriiformes orders had the most frequently sero-positive species to AIV. In conclusion, the presence of H5 and H9 subtypes provided a public health risk and evoke the role of migratory birds in introducing AIV to human and other in contact wild and domestic birds.