de Oliveira_2026_Molecules_31_

Aedes aegypti and Anopheles darlingi represent health challenges due to synthetic insecticide resistance. Hence, the essential oil from Piper humillimum is an alternative for vector control. In this study, the essential oil (3.5 +/- 0.4% yield) alongside germacrene D (61.51%) and delta-cadinene (17.46%) showed larvicidal activity (LC(50) of 34.75 to 46.04 microg/mL), accompanied by an increase of hydrogen peroxide (H(2)O(2)) production (36.67 +/- 1.52 to 81.33 +/- 1.52 micromol H(2)O(2) min(-1) mg(-1) protein), causing lipid (43.3 +/- 6.02 to 81.67 +/- 3.05 nmol malondialdehyde mg(-1) protein) and protein damages (61.67 +/- 6.80 to 83.00 +/- 2.64 nmol carbonyls mg(-1) protein). Further triggering an increase in superoxide dismutase (83.31 +/- 6.80 to 95.00 +/- 3.60 U mg(-1) protein) and catalase (74.31 +/- 7.02 to 82.09 +/- 1.00 micromol H(2)O(2) min(-1) mg(-1) protein) activities. In addition, mixed-function oxidases (61.17 +/- 11.37 to 73.52 +/- 6.42 nmol cyt c min(-1) mg(-1) protein), alpha- and beta-esterase (38.41 +/- 4.04 to 61.31 +/- 9.29 micromol min(-1) mg(-1) protein) levels increased. Conversely, glutathione S-transferase (GST) (11.01 +/- 2.00 to 9.67 +/- 3.05 micromol min(-1) mg(-1) protein) and acetylcholinesterase (AChE) (14.33 +/- 3.78 to 17.00 +/- 1.00 micromol min(-1) mg(-1) protein) were inhibited, corroborated by molecular docking, with germacrene D and delta-cadinene showing binding energies of -7.9 and -7.9 kcal/mol, 1.63 and 1.94 Ki for AChE, while for GST were -6.4 and 6.6 kcal/mol, and 20.5 and 15.50 Ki, respectively. These results demonstrate that the essential oil from P. humillimum is a promising multi-target alternative for the control of the investigated vectors.