Tissue specimens and serum samples obtained from adult budgerigars in various stages of reproduction housed in an aviary with enzootic avian polyomavirus (APV) disease were examined by means of polymerase chain reaction techniques for APV DNA. Although the birds were apparently healthy, APV DNA could be detected in all 40 birds examined (inapparent infection rate, 100%). Viral DNA was found in most organ systems examined. Analysis of data suggested that organ virus concentrations were lower in breeding than in nonbreeding birds. Serum samples from 144 birds were examined for virus-neutralizing (VN) antibody. All serum samples had detectable VN antibody titers. Determining VN titer had a sensitivity of 100% for detection of APV infection in birds and was more sensitive than analysis of droppings by use of polymerase chain reaction techniques to detect APV infection in 6-month-old birds. Analysis of the data suggested that lower VN antibody titers were associated with longer duration of continuous breeding.