Resistance to S-Methoprene Correlates with Pyriproxyfen Resistance in Field-Collected Culex pipiens

The increasing prevalence of insecticide resistance threatens the efficacy of Integrated Mosquito Management (IMM) programs, particularly in regions reliant on chemical control for vector-borne disease prevention. Cross-resistance between active ingredients severely complicates essential resistance management strategies like product rotation. The previous literature suggests that laboratory-induced S-methoprene-resistant Culex species may be somewhat cross-resistant to pyriproxyfen, another juvenile hormone analog. This is a critical concern in the Chicago, IL, USA metropolitan area, where pyriproxyfen is used against mosquitoes with reduced susceptibility to S-methoprene. To determine if S-methoprene-resistant Culex pipiens are cross-resistant to pyriproxyfen in nature, we assessed 31 field-collected populations with significant S-methoprene exposure but varying histories of pyriproxyfen use by dose-response bioassays. Culex pipiens from all 31 sites exhibited high resistance to S-methoprene (RR₅₀ > 10), and 84% were at least moderately resistant to pyriproxyfen (RR₅₀ > 5). Reduced susceptibility to pyriproxyfen was confirmed in pyriproxyfen-unexposed populations, demonstrating potential S-methoprene-mediated cross-resistance. The level of S-methoprene resistance and the level of pyriproxyfen exposure significantly correlated with the level of pyriproxyfen resistance. We report the first widespread, high-level pyriproxyfen resistance in any medically significant mosquito species, underscoring the critical need for routine resistance surveillance and the adoption of integrated resistance management tactics utilizing larvicides with distinct modes of action.