Objective To evaluate the feasibility of respiratory inhalation route on avian influenza virus(H5N1) in rhesus macaques.Method We inoculated rhesus macaques by thyrocricoid puncture and observed the clinical signs,pathologic changes and so on in a short period.Result Infection caused typical clinical signs and pathologic changes which were similar to human and animals infected with avian influenza virus.Conclusion Rhesus macaques could be infected by the inoculation of thyrocricoid puncture.And it was proved to be a good method for the similar experiments.