Abstract-Rhoicissus tomentosa is a plant used in South Africantraditional medicine to treat ailments mainly related to fertility and reproduction. Phytochemical screening was carried out and the antimicrobial activity of the rhizomes of R. tomentosa was investigated for the first time to ascertain their possible pharmaceutical potential. The results showed that the rhizomes of the plant contain alkaloids, flavonoids, saponins, steroids, reducing sugars and tannins. GCxGC-TOF-MS analysis of the extracts displayed the presence of over 100 known bioactive compounds, many reported here and detected in Rhoicissus tomentosa for the first time. Methanol: chloroform (50/50, v/v) and ethyl acetate (100%) extracts of the rhizomes were tested against 14 bacterial strains using the disc diffusion and microdilution assay methods. Bacteria most susceptible to rhizome extracts were Staphylococcus aureus (0.063 mg/mL) and Bacillus subtilis (0.125 mg/mL). The results obtained show that the rhizomes of R. tomentosa have antimicrobial activity against a number of pathogenic microorganisms.