From February to June 2006 an epidemic of highly pathogenic avian influenza (HPAI) virus Of subtype H5N1 affected all commercial poultry sectors as well as rural and backyard level in Egypt. This outbreak also was extended to include the zoo birds In Giza zoo. This study records the isolation and characterization of H5Nl virus from different species of the zoo birds and also studies the immune states of the vaccinated birds after application of H5Nl vaccine for the first time one month after the introduction of early outbreaks of 2006. Viral diagnosis was based on direct detection of viral RNA by real time PCR. Two positive cases from turkey and Grocer duck were processed for viral isolation and characterization. The level of antibodies was detected in vaccinated birds by HI test and the results were discussed to evaluate the role of vaccination in controlling the disease in these valuable zoo birds.