Highly pathogenic avian influenza (AI) disease has occurred in many countries globally adversely affecting domestic poultry production. Ghana recorded her first outbreak of a highly pathogenic avian influenza (HPAI) in 2007 on a small scale commercial farm in Tema. Since then, there have been numerous outbreaks. The source of these outbreaks is not conclusive. The role of wild birds in the epidemiology of avian influenza outbreaks in Ghana is not known. This study sought to investigate the role of wild birds in the outbreaks of Highly Pathogenic Avian Influenza (HPAI H5N1) in Ghana, particularly in Southern Ghana. Wild birds were trapped and sampled through mist netting. The faecal and tracheal samples were analysed using a One-Step Real Time Reverse Transcription Polymerase Chain reaction (RT-PCR) with primer sets targeting the matrix protein gene of the Avian influenza virus. Sera samples were subjected to multispecies competitive Enzyme Linked Immunosorbent Assay (ELISA) for anti-AI virus antibodies. Three hundred and twenty two (322) wild birds were trapped and sampled. Birds sampled included 87.3 % (281/322) resident birds and 12.7 % (41/322) migratory birds. The migratory birds included intra-African migrants 12.2 % (5/41) and Pale-arctic migrants 87.8 % (36/41). Avian influenza virus and antibody were neither detected in these swabs nor sera samples, respectively. The study documented the absence of AI in resident and migrant wild birds in the study area and suggest that wild birds may not be responsible for the outbreaks of AI in the poultry. However, sustained surveillance is recommended to ascertain a nationwide successful prevention and control strategy to stay the tide of any future intruding AI outbreaks.