Background: Sedum praealtumhas been used for a long time in traditional medicine as an analgesic and anti-inflammatory agent. Its beneficial effects have been known since ancient times, when Latinos used it to treat sore and swollen eyes. This research evaluated the antimicrobial potential, the cytotoxic and genotoxic effects, and some chromatographic profiles of the hydroethanolic extract of leaves, stems and roots ofS. praealtum.Methods: The antimicrobial activities were carried out by broth microdilution and agar diffusion. In vitro cytotoxicity was evaluated by cell cultures ofAedes albopictusand the selectivity index (SI) was estimated: SI=CI50/MIC. Genotoxic and systemic toxic effects ofS. praealtumleaves were analyzed by micronucleus assay in mice bone marrow. Chromatographic profiles and mass spectra were investigated by GC-MS.Results: Gram-positive (B. subtilis,B. cereus,M. luteus,E. faecalisandS. aureus) and gram-negative (E. coli,E. aerogenes,S. marcescens,P. aeruginosa,P. mirabilisandS. typhimurium) bacteria exhibited MICs ranging from 12.5–50 and 0–50 mg/ml, respectively.Sedum praealtumshowed no efficacy againstM. tuberculosisandM. bovis. Cytotoxicity (CI50) ofS. praealtumwas 4.22 and 5.96 mg/ml for leaves and stems, respectively, while its roots showed no cytotoxicity. Micronucleated polychromatic erythrocytes (MNPCEs) analyzes showed no differences between treatment doses (0.5–2 g/kg) and negative control (NaCl), but the PCE/NCE ratio (polychromatic erythrocyte/normochromatic erythrocyte) showed significant differences. Phytochemical screening identified thirteen compounds in the leaves, stems and roots ofS. praealtumpotentially associated with their biological activities.Conclusions: This research comprises a first scientific study on genotoxicity, cytotoxicity and antimicrobial effects ofS. praealtum(Balsam), and it provides an initial theoretical foundation for its comprehensive use. Results showed antibacterial action ofS. praealtumagainst gram-positive bacteria and some gram-negative species (depending on the plant anatomical part), but ineffective antimycobacterial action. However,S. praealtumleaves and stems display potential cytotoxicity, contributing to the SI < 1 values. In addition,S. praealtumleaves exhibit no clastogenic and/or aneugenic effects, but it has systemic toxicity dose-independent.