The effect of suramin on duck hepatitis B virus (DHBV) infection was investigated in vivo. Suramin pretreatment of Pekin ducklings completely prevented DHBV infection. In contrast, suramin given at the time of or after inoculation with DHBV did not inhibit viral infection, replication, or gene expression. These data indicate that suramin effectively blocks the early stages of DHBV infection in vivo.Suramin, a polysulfonated naphthylurea, was the first clinically useful antiparasitic drug and is still one of the most commonly used antitrypanosomal agents. Apart from its antiparasitic effects, suramin has been demonstrated to have a number of biochemical activities (7). Most notably, suramin has been shown to inhibit a number of enzymes, such as hexokinase, lysozyme (11), thrombin, plasma kallikrein (5), and protein kinase C (6). Since suramin was shown to bind to a number of growth factors (8) and protein kinase C is involved in tumor promotion and differentiation, the antitumor activity of suramin has been studied in patients with metastatic adrenocortical carcinoma (18) and metastatic prostate cancer (9). In addition to its antiparasitic and antitumor properties, suramin has been shown to strongly inhibit duck hepatitis B virus (DHBV) DNA polymerase (19,20) and the reverse transcriptase activities of a number of retroviruses (3), including the human immunodeficiency virus (13). A clinical trial in 10 patients with AIDS demonstrated that suramin indeed inhibits human immunodeficiency virus replication; immunological parameters and clinical outcomes were not affected, however (1).After the discovery of DHBV (12), Pekin ducks have become a widely used animal model to study hepadnaviral infection. By using this model, antiviral agents can be studied in vitro in replicative complexes isolated from DHBV-infected livers (16) or in primary duck hepatocyte cultures (18) as well as in vivo in DHBV-infected Pekin ducks (15). In primary duck hepatocyte cultures, suramin blocks infection when it is present before and during incubation of the cells with DHBV (17). Since suramin inhibits DHBV infection in vitro, we tested the effect of suramin on DHBV infection in vivo.Pekin ducks were used as an in vivo model of DHBV infection. One-day-old ducklings were purchased from a German duck farm. Blood was taken from the foot vein and was tested for the presence of DHBV DNA by dot blot hybridization (14). DHBV DNA-negative ducklings were infected at the age of 2 or 3 days by intravenous injection of 100 ,ul of DHBV-positive serum (about 109 virions per ml).