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Routine monitoring of occurrence, levels and mechanisms of insecticide resistance informs effective management strategies, and should be used to assess the effect of new tools on resistance. As part of a cluster randomised controlled trial evaluating a novel insecticide-based intervention in central Côte d’Ivoire, we assessed resistance and its underlying mechanisms in  Anopheles gambiae  populations from a subset of trial villages. Resistance to multiple insecticides in  An. gambiae s.s.  and  An. coluzzii  was detected across villages, with dose–response assays demonstrating extremely high resistance intensity to the pyrethroid deltamethrin (> 1,500-fold), and mortality following exposure to pyrethroid-treated bednets was low (< 30% mortality in cone bioassays). The 1014F  kdr  mutation was almost fixed (≥ 90%) in all villages but the 1575Y  kdr -amplifying mutation was relatively rare (< 15%). The carbamate and organophosphate resistance-associated  Ace-1  G119S mutation was also detected at moderate frequencies (22–43%). Transcriptome analysis identified overexpression of P450 genes known to confer pyrethroid resistance ( Cyp9K1 ,  Cyp6P3 , and  Cyp6M2 ), and also a carboxylesterase ( COEAE1F ) as major candidates.  Cyp6P3  expression was high but variable (up to 33-fold) and correlated positively with deltamethrin resistance intensity across villages (r 2  = 0.78, P = 0.02). Tools and strategies to mitigate the extreme and multiple resistance provided by these mechanisms are required in this area to avoid future control failures.

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Fine scale spatial investigation of multiple insecticide resistance and underlying target-site and metabolic mechanisms in Anopheles gambiae in central Côte d’Ivoire