The aim of this study was to evaluate the biological potential of the crude hydroalcoholic extract (EBH) and fractions (FAcOEt and FBuOH) from the roots of Dioscorea scabra. The EBH and its fractions were submitted to antimicrobial activity tests against bacteria with and without cell-wall, and yeasts from the Candida genus. The phytochemical profile of EBH was determined by colorimetric assays and analysis by HPLC in comparison to standard diosgenin. Phytochemical analysis showed the presence of phenols, tannins, flavonoids, xanthones, catechins, steroids, free triterpenoids and saponins, characteristics of the genus. By HPLC analysis, it was possible to identify the presence of diosgenin in the analysed extract. The species was active against all tested bacterial strains, being FBuOH the most active against Mycoplasma arginini species. In the antifungal activity test, samples showed activity against Candida albicans, C. parapsilosis and C. glabrata, with the EBH and FBuOH showing the best results.