Abstract Anti-hepatitis B viral activity of ethanol stem bark extracts of Vitex doniana and Lophira alata in Wistar rats was investigated. Forty-two (42) humanized albino rats were used for the study. The animals were divided into six (6) groups of seven (7) animals each. Groups II-VI were inoculated with hepatitis-B virus at the concentration of 25μl of the virus preparation was inoculated into a day old suckling humanized Wistar rat via intra cerebral route using tuberculin syringe. Group 1 and 2 served as normal and positive controls respectively and were given Dimethylsulphoxide/distilled water as placebo. Group III received 300mg/kgBW of standard anti-retroviral drug (Tenofovir-Lamivudin), group IV received 244mg/kgBW of Vitex doniana, group V received 244mg/kgBW of Lophira alata and group VI received combination of Vitex doniana and Lophira alata both at 244mg/kgBW. The treatment was administered twice daily for ninety days with the aid of orogastric tube. Result of the study reveal a drastic reduction in viral load in all treated groups; groups III (172.47 ± 3.99), IV (18.58 ± 1.01), V (548.48 ± 3.26) and VI (0.00 ± 0.00) with more significant (p<0.05) reduction in the combined treated group; group VI (0.00 ± 0.00) which recorded no significant (p<0.05) differences and compared well with the normal control (0.00 ± 0.00). Results of biochemical indices determination revealed a significant (p<0.05) increase in AST activity in group VI compared with normal controls but was significantly lowered when compared with positive control. However, ALT activities in group V showed a significant (p<0.05) decrease while that of group VI was significantly (p<0.05) increased compared with normal controls. ALP activities showed significant (p<0.05) reduction in all treatment groups compared with the positive control. Total protein, globulin and albumin showed significant (p<0.05) increased in all treated groups compared with the positive controls. The results obtained for the above parameters in all treated groups may be as a result of the administration of the extracts which reduces the HBV viral load and hence, reversing hepatic necrosis and morphological distortion occasion by viral toxins on the liver tissues. Co-administration of the two plants extracts in the treatment of HBV-induced hepatitis in Wistar rats appears to be effective, safe at the dose investigated compared with the standard drug (Tenofovir-Lamivudin) and can be a candidate for further research and development new anti-viral drug.