Diseases transmitted by blood-feeding mosquitoes, such as dengue fever, dengue hemorrhagic fever, Japanese encephalitis, malaria, and filariasis, are increasing in prevalence, particularly in tropical and subtropical zones. To control mosquitoes and mosquito-borne diseases, which have a worldwide health and economic impacts, synthetic insecticide-based interventions are still necessary, particularly in situations of epidemic outbreak and sudden increases of adult mosquitoes. However, the indiscriminate use of conventional insecticides is fostering multifarious problems like widespread development of insecticide resistance, toxic hazards to mammals, undesirable effects on nontarget organisms, and environmental pollution. The aim of this research was to evaluate the toxicity of mosquito larvicidal activity of essential oil from Coleus aromaticus and its pure isolated constituent thymol against larvae of Culex tritaeniorhynchus, Aedes albopictus, and Anopheles subpictus. The chemical composition of the essential oil was analyzed using gas chromatography-mass spectroscopy. A total of 14 components of the essential oil of C. aromaticus were identified. The major chemical components identified were thymol (82.68%), terpinen-4-ol (3.2%), and trans-Caryophyllene (3.18%). Twenty-five early third instar larvae of C. tritaeniorhynchus, A. albopictus, and A. subpictus were exposed and assayed in the laboratory. Thymol and essential oil were tested in concentrations of 10, 20, 30, 40, and 50 and 30, 60, 90, 120, and 150 ppm, respectively. The larval mortality was observed after 24 h of treatment. The thymol had a significant toxic effect against early third-stage larvae of C. tritaeniorhynchus, A. albopictus, and A. subpictus with an LC50 values of 28.19, 24.83, and 22.06 μg/mL respectively, whereas the essential oil of C. aromaticus had an LC50 values of 72.70, 67.98, and 60.31 μg/mL, respectively. No mortality was observed in controls. The Chi-square values were significant at p < 0.05 level. The result indicated that the essential oil of C. aromaticus and the isolated constituent have a potential for use in control of C. tritaeniorhynchus, A. albopictus, and A. subpictus larvae and could be useful in search of newer, safer, and more effective natural compounds as larvicides.