Aedes aegypti is the principal mosquito vector for many arthropod-borne viruses (arboviruses) including dengue, chikungunya, and Zika. In the United States, excessive permethrin use has led to a high frequency of resistance in mosquitoes. Insecticide resistance is a significant obstacle in the struggle against vector-borne diseases. To help overcome metabolic resistance, synergists that inhibit specific metabolic enzymes can be added to formulated pyrethroid products. Using modified CDC bottle bioassays, we assessed the effect of three inhibitors (piperonyl butoxide (PBO), which inhibits oxidase activity; S.S.S-tributyl phosphorotrithioate (DEF), which inhibits esterase activity; and diethyl maleate (DM), which inhibits glutathione transferase activity) + permethrin. We performed these against 20 Florida Ae. aegypti populations, all of which were resistant to permethrin. Our data indicated that 11 out of 20 populations (55%) exhibited metabolic resistance. Results revealed 73% of these populations had significant increases in mortality attributed to DEF + permethrin, 64% to PBO + permethrin, and 55% to DM + permethrin compared to permethrin alone. Currently, PBO is the only metabolic enzyme inhibitor added to formulated pyrethroid products used for adult mosquito control. Our results suggest that the DEF and DM inhibitors could also be useful additives in permethrin products, especially against metabolically resistant Ae. aegypti mosquitoes. Moreover, metabolic assays should be conducted to better inform mosquito control programs for designing and implementing integrated vector management strategies.

Insecticide resistance in Aedes aegypti (Linnaeus, 1762) poses a major challenge for vector control programs, undermining the effectiveness of chemical tools to mitigate both nuisance biting and the transmission of vector-borne diseases. Previous studies have documented widespread resistance to multiple adulticide active ingredients across Florida, particularly pyrethroids, along with high prevalence of knockdown resistance (kdr) mutations. Continued resistance monitoring is essential for guiding treatment decisions and detecting shifts in susceptibility over time. In collaboration with mosquito control programs across Florida, we assessed the susceptibility of 78 Ae. aegypti populations to up to six active ingredients, conducting 396 independent CDC bottle bioassays and calculating hazard ratios relative to a susceptible Ae. aegypti strain. For pyrethroids, 24 h post-exposure mortality was used to evaluate phenotypic recovery from knockdown. Hazard ratios revealed Ae. aegypti populations were more susceptible to organophosphate adulticide active ingredients, while the lowest hazard ratios, indicating higher resistance, were observed for sumithrin, deltamethrin, etofenprox, and permethrin, respectively. Evidence of knockdown resistance and recovery after 24 h was present across all pyrethroid exposures, with the highest levels following treatment with etofenprox and sumithrin. These findings confirm that pyrethroid resistance remains a significant barrier to Ae. aegypti control and provide updated, operationally relevant resistance data that can complement existing guidance and support evidence-based vector management strategies.