Abstract

BackgroundThe Democratic Republic of the Congo (DRC) is characterized as a holoendemic malaria area with the main vectors being Anopheles funestus and members of the Anopheles gambiae complex. Due to political instability and socio-economic challenges in the region, knowledge of insecticide resistance status and resistance mechanisms in these vectors is limited. Mosquitoes were collected from a mining site in the north-eastern part of the country and, following identification, were subjected to extensive testing for the target-site and biochemical basis of resistance. Quantitative real-time PCR was used to assess a suite of 10 genes frequently involved in pyrethroid and dichlorodiphenyltrichloroethane (DDT) resistance in An. gambiae females and males. In An. funestus, gene expression microarray analysis was carried out on female mosquitoes.ResultsIn both species, deltamethrin resistance was recorded along with high resistance and suspected resistance to DDT in An. gambiae and An. funestus, respectively. A total of 85% of An. gambiae carried the kdr mutations as either homozygous resistant (RR) (L1014S, L1014F or both) or heterozygous (RS), however only 3% carried the rdl mutant allele (RS) and no ace-1 mutations were recorded. Synergist assays indicated a strong role for P450s in deltamethrin resistance in both species. In An. gambiae, analysis of transcription levels showed that the glutathione-S-transferase, GSTS1-2, produced the highest fold change in expression (7.6-fold in females and 31-fold in males) followed by GSTE2, thioredoxin peroxidase (TPX2), and cytochrome oxidases (CYP6M2 and CYP6P1). All other genes tested produced fold change values below 2. Microarray analysis revealed significant over-transcription of cuticular proteins as well as CYP6M7, CYP6P9a and CYP6P9b in insecticide resistant An. funestus.ConclusionsThese data show that high levels of deltamethrin resistance in the main malaria vector species, conferred by enzymatic detoxification, are present in the DRC.

Highlights

  • The Democratic Republic of the Congo (DRC) is characterized as a holoendemic malaria area with the main vectors being Anopheles funestus and members of the Anopheles gambiae complex

  • Vector control in the DRC is largely based on the use of long-lasting insecticide treated bed nets (LLINs) [3, 4], and to a lesser extent, indoor residual spraying (IRS) which is limited to areas where mining operations are present [5]

  • There has been a strong reliance on the pyrethroids as they are the only class of insecticide approved for treating bed nets, which have been central to reducing malaria prevalence

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Summary

Introduction

The Democratic Republic of the Congo (DRC) is characterized as a holoendemic malaria area with the main vectors being Anopheles funestus and members of the Anopheles gambiae complex. Vector control in the DRC is largely based on the use of long-lasting insecticide treated bed nets (LLINs) [3, 4], and to a lesser extent, indoor residual spraying (IRS) which is limited to areas where mining operations are present [5]. Pyrethroids, organochlorines, carbamates and organophosphates are the only four classes of insecticide approved for use in vector control [6]. There has been a strong reliance on the pyrethroids as they are the only class of insecticide approved for treating bed nets, which have been central to reducing malaria prevalence.

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