Insecticide resistance in Anopheles mosquitoes is seriously threatening the success of insecticide-based malaria vector control. Surveillance of insecticide resistance in mosquito populations and identifying the underlying mechanisms enables optimisation of vector control strategies. Here, we investigated the molecular mechanisms of insecticide resistance in three Anopheles coluzzii field populations from southern Côte d’Ivoire, including Agboville, Dabou and Tiassalé. All three populations were resistant to bendiocarb, deltamethrin and DDT, but not or only very weakly resistant to malathion. The absence of malathion resistance is an unexpected result because we found the acetylcholinesterase mutation Ace1-G280S at high frequencies, which would typically confer cross-resistance to carbamates and organophosphates, including malathion. Notably, Tiassalé was the most susceptible population to malathion while being the most resistant one to the pyrethroid deltamethrin. The resistance ratio to deltamethrin between Tiassalé and the laboratory reference colony was 1,800 fold. By sequencing the transcriptome of individual mosquitoes, we found numerous cytochrome P450-dependent monooxygenases – including CYP6M2, CYP6P2, CYP6P3, CYP6P4 and CYP6P5 – overexpressed in all three field populations. This could be an indication for negative cross-resistance caused by overexpression of pyrethroid-detoxifying cytochrome P450s that may activate pro-insecticides, thereby increasing malathion susceptibility. In addition to the P450s, we found several overexpressed carboxylesterases, glutathione S-transferases and other candidates putatively involved in insecticide resistance.

Background: In order to assess the impact of the different groups of agricultural pesticides used in Côte d'Ivoire on the increase of mosquitoes resistance to insecticides, the expression profiles of 7 P450 cytochromes and one GSTE2 of Anopheles gambiae involved in mosquito resistance to insecticides were studied. The goal of this study was to determine the effect of short exposure of mosquito larvae to different groups of agricultural pesticides on mosquito resistance. Methods: Three groups of pesticides were selected: (i) agricultural insecticide solutions, (ii) non-insecticide pesticide solutions (a mixture of herbicides and fungicides), and (iii) a mixture of the first two. A fourth non-pesticide solution was used as a control. Four groups of each stage 2 larvae (strain Kisumu, male and female) were exposed to 20% concentrated solution for 24 hours. Susceptibility tests for dichlorodiphenyltrichloroethane (DDT) and Deltamethrin were carried out on adults aged 2-5 days. Quantitative reverse transcription polymerase chain reaction (RT-qPCR) was performed to quantify the expression of eight metabolic genes involved in mosquito resistance to insecticides.  Results: Susceptibility to DDT showed a similar increase in the time required to knock down 50% of mosquitoes (kdt50) in l colonies exposed to insecticides and non-insecticides compared to the control colony. As for deltamethrin, kdt50 was higher in the colonies exposed to insecticides and the pesticide mixture compared to the colony exposed to non-insecticides. Of all the genes studied in all colonies, except for CYP6P1 induced only in the colony consisting of the pesticide mixture, no genes were induced. Conclusions: This study confirmed that induction is influenced by the duration, the concentration of the solution and the type of xenobiotic used as an inducer. The overexpression of CYP6P1 confirmed the inductive effect that a short exposure of mosquito larvae to agricultural pesticides could have.