Botanical insecticides are preferred for their environment and user-friendly nature. Eugenol is a plant-based monoterpene having multifarious biocidal activities. To understand whether eugenol would persistently work against Aedes aegypti, we performed larvicidal bioassays on thirty successive generations and determined median lethal concentration (LC50) on each generation. Results showed no apparent differences between LC50 at F0 (63.48 ppm) and F30 (64.50 ppm) indicating no alteration of susceptibility toward eugenol. To analyze, if eugenol has any effect on metabolic detoxification-associated enzymes, we measured esterases (alpha and beta), cytochrome P450, and GST activities from the survived larvae exposed to LC50 concentration from F0-F30. Results revealed a decrease of esterases, GST, and cytochrome P450 activities at the initial 4-8 generations and then a gradual increase as the generations progressed. GST activity remained significantly below the control groups. Synergists (TPP, DEM, and PBO) were applied along with eugenol at F30 and LC50 concentration, and the said enzyme activities were recorded. Results showed a noticeable decrease in LC50 and enzyme activities indicating effective inhibitions of the respective enzymes. Overall, present results inferred that eugenol would effectively work as a larvicide for a longer period in successive generations without initiating rapid resistance and therefore could be advocated for controlling A. aegypti.
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