Entada abyssinica, Securinega virosa and Ehretia amoena are traditionally used in Southern Uganda to treat Human African Trypanosomiasis (HAT). Extracts from the roots and root barks of E. abyssinica, S. virosa and E. amoena were investigated for in vivo antitrypanosomal activity against Trypanosoma brucei brucei in mice. The extracts were also tested in vitro for potential cytotoxicity against a 3-cell line panel consisting of TK10 (renal), UACC62 (melanoma) and MCF7 (breast) cancer cells using the highly sensitive Sulforhodamine B assay. The ethanolic extract from the roots of E. abyssinica showed a reduction of T. b. brucei parasitaemia at a single dose of 400 mg/kg. The petroleum ether extract from the root bark of E. abyssinica showed the highest cytotoxicity with GI50, total growth inhibition, LC50 and LC100 values of less than 6.25 µg/ml. Our findings confirm earlier in vitro antitrypanosomal studies and lend credence to the traditional use of these plants against trypanosomiasis.