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Purpose: To evaluate the larvicidal activity of the essential oil of Dipsacus asper Wallich ex Candolle (Dipsacaceae) roots against the larvae of Aedes aegypti L. and Culex pipiens pallens Coquillett.Methods: Essential oil was extracted from D. asper roots by hydrodistillation and analyzed for its composition by gas chromatography (GC) and gas chromatography-mass spectrometry (GC-MS). The oil was evaluated for larvicidal activity, using World Health Organization (WHO) procedures, against the fourth larvae of A. aegypti and C. pipiens pallens within 24 h, and activity was recorded for various concentrations of the oil ranging from 12.5 – 200.0 μg/mL.Results: A total of 34 components of the essential oil of D. asper were identified. The major compounds of the essential oil were caryophyllene oxide (13.29 %), caryophyllene (9.14 %), cubebene (7.87 %), β-gurjunene (6.43 %), carvone (5.38 %), 1,8-cineole (5.29 %), and calamenene (5.05 %). The oil exhibited larvicidal activity against A. aegypti and C. pipiens pallens at median lethal concentrations (LC50) of 56.29 μg/mL and 47.49 μg/mL, respectively.Conclusion: The essential oil of D. asper roots has potentials for use in the control of A. aegypti and C. pipiens pallens and may be useful in the search for new, safer and more effective natural larvicides.Keywords: Dipsacus asper, Aedes aegypti, Culex pipiens pallens, Essential oil, Larvicidal activity, Caryophyllene, Cubebene, β-Gurjunene, Carvone