The methanolic extract (DAT), fractions (FRS) and four flavonoids, namely Gancaonin Q (1), Stipulin (2), Angusticornin B (3) and Bartericin A (4), isolated from the twigs of Dorstenia angusticornis (Moraceae), were tested for their in vitro antimicrobial activity. A total of 22 microbial cultures belonging to three Candida species, 6 Gram-positive and 13 Gram-negative bacterial species were used in this study. The inhibition zones (IZ) of the test samples against the pathogens were determined by the Agar Hole Diffusion test while the Liquid dilution method was used to determine their minimal inhibition concentrations (MIC) and their minimal microbicidal concentrations (MMC). Results indicate that DAT, compounds 3 and 4 inhibited the growth of all test pathogens. DAT, FRS 3-6, compounds 3 and 4 were both antibacterial and anticandidal. A single-dose oral toxicity performed in accordance with the OPPTS 870.1100 and OECD 401guideline showed that DAT was not toxic. Our findings provide a possible basis for the potential use of twigs from Dorstenia angusticornis in the treatment of infectious diseases.