Biochemical mechanisms of insecticide resistance of thermal exposed and unexposed Culex quinquefasciatus strains are evaluated, which were not studied earlier. The activity of alpha- and beta-carboxylesterases and acetylcholinesterase of malathion susceptible and resistant strains were compared after thermal treatment. Three-day-old adult females were used for the malathion susceptibility test and biochemical assays, and males were used only for the susceptibility test. Thermal exposure brought about increase in resistance levels from 85% to 90% in males and 91% to 96.6% in females of resistant strain. The resistance status of the susceptibility strain was unchanged after thermal exposure. The activities of alpha- and beta-carboxylesterase of susceptible mosquitoes were within 800 and 700 U/mg protein, respectively. The alpha-carboxylesterase activity of the thermal exposed malathion-resistant population was significantly (t test, P < 0.05) higher than the unexposed resistant population, and the reverse was recorded in beta-carboxylesterase. The alpha-carboxylesterase activity of susceptible population was lower than the resistant population. The activity of alpha-carboxylesterase was higher than the beta-carboxylesterase in both the strains. Among the malathion resistant C. quinquefasciatus population, 2.3% population exhibited 30-40% inhibition which increased to 5.8% after the thermal exposure. Thermal exposure of mosquitoes increased the activity of both alpha-carboxylesterases and acetylcholinesterase but decreased the activity of beta-carboxylesterase.