Human cases of disease caused by highly pathogenic avian influenza (HPAI) viruses of the H5N1 subtype are rare, yet characterized with a mortality rate of approximately 60%. Tests were conducted to determine the environmental persistence of an HPAI (H5N1) virus on four materials (glass, wood, galvanized metal, and topsoil) that could act as fomites or harbor the virus. Test coupons were inoculated with the virus and exposed to one of five environmental conditions that included changes in temperature, relative humidity, and simulated sunlight. At time periods up to 13 days, the virus was extracted from each coupon, and quantified via cytopathic effects on Madin-Darby canine kidney cells. The virus was most persistent under the low temperature condition, with less than 1 log reduction on glass and steel after 13 days at low relative humidity. Thus, at these conditions, the virus would be expected to persist appreciably beyond 13 days.