| Summary: | This work aimed to prepare a nanoemulsion containing the essential oil of the <i>Protium heptaphyllum</i> resin and evaluate its biocidal activities against the different stages of development of the <i>Aedes aegypti</i> mosquito. Ovicide, pupicide, adulticide and repellency assays were performed. The main constituents were <i>p</i>-cymene (27.70%) and <i>α</i>-pinene (22.31%). The developed nanoemulsion showed kinetic stability and monomodal distribution at a hydrophilic–lipophilic balance of 14 with a droplet size of 115.56 ± 1.68 nn and a zeta potential of −29.63 ± 3.46 mV. The nanoemulsion showed insecticidal action with LC<sub>50</sub> 0.404 µg·mL<sup>−1</sup> for the ovicidal effect. In the pupicidal test, at the concentration of 160 µg·mL<sup>−1</sup>, 100% mortality was reached after 24 h. For adulticidal activity, a diagnostic concentration of 200 µg·mL<sup>−1</sup> (120 min) was determined. In the repellency test, a concentration of 200 µg·mL<sup>−1</sup> during the 180 min of the test showed a protection index of 77.67%. In conclusion, the nanobiotechnological product derived from the essential oil of <i>P. heptaphyllum</i> resin can be considered as a promising colloid that can be used to control infectious disease vectors through a wide range of possible modes of applications, probably as this bioactive delivery system may allow the optimal effect of the <i>P. heptaphyllum</i> terpenes in aqueous media and may also induce satisfactory delivery to air interfaces.
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