Thirty-five budgerigars were infected with the Psittacid herpesvirus 1 (RSL-1 strain, ATCC) to study the pathogenesis of Pacheco's disease. Intramuscular (i.m.) and oral (p.o.) infection routes were used in 21 and 14 animals respectively. Animals were euthanized on days 1, 2, 3, 4, 6 and 8 post-inoculation (p.i.) and complete postmortem examinations and histological studies were performed. The presence of viral antigen in tissues was detected by immunohistochemical techniques using a rabbit polyclonal antibody. In the i.m.-infected birds, lesions were first detected on the third day p.i. and included necrotizing hepatitis and splenitis, both associated to the presence of viral antigen. Necrotic and inflammatory lesions as well as viral antigen were detected in many organs after the fourth day p.i. (oesophagus, crop, pancreas, kidney, adrenal gland, thyroid and parathyroid glands, thymus, ovary and feathers) proving generalization of the disease. Chronology of the infection was similar in the p.o.-infected birds. However, two main differences were observed between the groups: In the p.o. group, viral antigen was first detected in cloacal mucosa (3rd day p.i.), liver and spleen; and viral entry into target cells and dissemination to the rest of the tissues was slower in this group. In addition, detection of viral antigen in feather follicular epithelial cells and in granulosa cells of organ follicles are findings that could be of relevance to the transmission of the virus.