The survival of avian pneumovirus (APV) in turkey litter was studied at different temperature (room temperature, [approximately 22–25 C], 8 C, and −12 C) conditions. Built-up turkey litter from a turkey breeder farm known to be free of APV was obtained and was divided into two portions. One portion was sterilized by autoclaving and the other portion was kept nonautoclaved. Both samples were inoculated with a Vero cell–propagated Minnesota isolate of APV subtype C (APV/MN2A) with a titer of 10⁵ 50% tissue culture infective dose at 1% level. These samples were then stored at three different temperatures: −12 C, 8 C, and room temperature (20–25 C). The samples were tested for the presence of viral RNA by reverse transcriptase–polymerase chain reaction and for the presence of live virus by virus isolation in Vero cells at the intervals of 1, 2, 3, 7, 14, 30, 60, and 90 days. Our studies revealed the presence of APV RNA even after 90 days in the autoclaved litter samples kept at −12 C and at 8 C. The virus was isolated from the autoclaved litter kept at −12 C up to 60 days. From the nonautoclaved litter, viral RNA was detected up to 60 days and virus was isolated up to 14 days. The present study indicated that APV could survive in built-up turkey litter up to 60 days postinoculation at a temperature of −12 C.