Twenty-nine Pekin ducks were inoculated with duck hepatitis B virus (DHBV), DHBV-free serum, or saline at 1 day of age. Congenitally DHBV-infected ducks were also studied. Ducks were killed periodically during a 92-week study and examined histologically and immunohistochemically to assess liver and extrahepatic inflammation and to detect and characterize DHBV core antigen tissue distribution. DHBV infection produced an asymptomatic but persistent DHBV viremia in all ducks associated with a mild to moderate transient hepatic inflammation apparent at 3 to 6 weeks post-inoculation and waning afterwards. DHBV core antigen was detected in hepatocyte cytoplasm at 1 week post-inoculation, and by 3 weeks post-inoculation scattered pancreatic acinar and islet cells also contained viral antigen. Small numbers of mononuclear cells in the splenic white pulp also contained viral antigen. Viral antigen persisted in all of these tissues throughout the duration of the experiment. No inflammation or tissue injury was detected in any of the extrahepatic tissues during the course of DHBV infection. One DHBV-injected duck developed a hepatocellular carcinoma at 88 weeks of age. Isolated patches of neoplastic hepatocytes contained cytoplasmic DHBV core antigen. The results of this study indicate that DHBV, like mammalian hepadnavirus, is capable of producing a persistent infection of the liver and several extrahepatic tissues and suggest that persistent infection may be associated with the development of hepatocellular carcinoma.