Objective: In this work, we evaluated the chemical composition by chromatography coupled to mass spectrometry, antimicrobial and insecticidal activity of essential oils (TrEO) and nanoeffu-sion (NTrEO) of oil from Tetradenia riparia leaves. We also evaluated the acute toxicity in Mus musculus. The results showed that S. aureus showed greater susceptibility to TrEO, with a minimum inhibitory concentration (MIC) of 125 µg.mL-1, when compared to P. aeruginosa, E. coli, which presented a MIC of 250 µg.mL-1. There was a greater toxic effect on TrEO (LC50 = 630.29 µg.mL-1) and NTrEO (LC50 = 713.68 µg.mL-1) larvae, effect on pupae (LC50 = 630.29 µg.mL-1) and NTrEO (LC50 = 713.68 µg.mL-1) and ovicide (LC50 = 631.184 µg.mL-1) and NTrEO (LC50 = 146.701 µg.mL-1) of Ae. aegypti after 24 h of exposure, when compared to other exposure periods. Chemical analysis re-vealed 20 components, aromadendrene oxide (23.47%) being the main component. Female and male mice were used to assess toxicity and oral administration of NTrEO to mice did not result in acute toxicity. Thus, the essential oil of T. riparia and its nanoemulsion have insecticidal and an-timicrobial potential for Ae. aegypti, S. aureus, E. coli, P. aeruginosa, and has a high toxicological safety threshold at the concentrations evaluated in this study.