title: The recent escalation in strength of pyrethroid resistance in Anopheles coluzzi in West Africa is linked to increased expression of multiple gene families.
abstract: BACKGROUND
  Since 2011, the level of pyrethroid resistance in the major malaria mosquito, Anopheles coluzzi, has increased to such an extent in Burkina Faso that none of the long lasting insecticide treated nets (LLINs) currently in use throughout the country kill the local mosquito vectors. We investigated whether this observed increase was associated with transcriptional changes in field-caught Anopheles coluzzi using two independent whole-genome microarray studies, performed in 2011 and 2012.
  
  
  RESULTS
  Mosquitoes were collected from south-west Burkina Faso in 2011 and 2012 and insecticide exposed or non-exposed insects were compared to laboratory susceptible colonies using whole-genome microarrays. Using a stringent filtering process we identified 136 genes, including the well-studied detoxification enzymes (p450 monoxygenases and esterases) and non-detoxification genes (e.g. cell transporters and cuticular components), associated with pyrethroid resistance, whose basal expression level increased during the timeframe of the study. A subset of these were validated by qPCR using samples from two study sites, collected over 3 years and marked increases in expression were observed each year. We hypothesise that these genes are contributing to this rapidly increasing resistance phenotype in An. coluzzi. A comprehensive analysis of the knockdown resistance (kdr) mutations (L1014S, L1014F and N1575Y) revealed that the majority of the resistance phenotype is not explained by target-site modifications.
  
  
  CONCLUSIONS
  Our data indicate that the recent and rapid increase in pyrethroid resistance observed in south-west Burkina Faso is associated with gene expression profiles described here. Over a third of these candidates are also overexpressed in multiple pyrethroid resistant populations of An. coluzzi from neighbouring Côte d'Ivoire. This suite of molecular markers can be used to track the spread of the extreme pyrethroid resistance phenotype that is sweeping through West Africa and to determine the functional basis of this trait.
authors.affiliation: Department of Vector Biology, Liverpool School of Tropical Medicine, Liverpool, L3 5QA, UK. H.Toe@liverpool.ac.uk.|Centre National de Recherche et de la Formation sur le Paludisme, Ouagadougou, 01BP 2208, Burkina Faso. H.Toe@liverpool.ac.uk.
authors.forename: Kobié H
authors.initials: KH
authors.lastname: Toé
authors.affiliation: Centre National de Recherche et de la Formation sur le Paludisme, Ouagadougou, 01BP 2208, Burkina Faso. n.fale.cnlp@fasonet.bf.
authors.forename: Sagnon
authors.initials: S
authors.lastname: N'Falé
authors.affiliation: Institut de Recherche en Sciences de la Santé/Centre Muraz, Bobo-Dioulasso, 01 BP 545, Burkina Faso. dabire_roch@hotmail.com.
authors.forename: Roch K
authors.initials: RK
authors.lastname: Dabiré
authors.affiliation: Department of Vector Biology, Liverpool School of Tropical Medicine, Liverpool, L3 5QA, UK. Hilary.Ranson@lstmed.ac.uk.
authors.forename: Hilary
authors.initials: H
authors.lastname: Ranson
authors.affiliation: Department of Vector Biology, Liverpool School of Tropical Medicine, Liverpool, L3 5QA, UK. christopher.jones@rothamsted.ac.uk.
authors.forename: Christopher M
authors.initials: CM
authors.lastname: Jones
mesh_terms: D000354:Africa, Western; D000818:Animals; D000852:Anopheles / Q000187:drug effects / Q000235:genetics*; D020346:Aquaporins / Q000235:genetics / Q000378:metabolism; D002050:Burkina Faso; D002265:Carboxylic Ester Hydrolases / Q000235:genetics / Q000378:metabolism; D016000:Cluster Analysis; D004351:Drug Resistance / Q000235:genetics; D016678:Genome; D005838:Genotype; D005982:Glutathione Transferase / Q000235:genetics / Q000378:metabolism; D006239:Haplotypes; D007306:Insecticides / Q000633:toxicity; D006899:Mixed Function Oxygenases / Q000235:genetics / Q000378:metabolism; D005810:Multigene Family; D009570:Nitriles / Q000633:toxicity; D020411:Oligonucleotide Array Sequence Analysis; D011722:Pyrethrins / Q000633:toxicity; D012313:RNA / Q000302:isolation & purification / Q000378:metabolism; D060888:Real-Time Polymerase Chain Reaction; D059467:Transcriptome
chemical_list: D020346:Aquaporins; D007306:Insecticides; D009570:Nitriles; D011722:Pyrethrins; C017180:decamethrin; D012313:RNA; D006899:Mixed Function Oxygenases; D005982:Glutathione Transferase; D002265:Carboxylic Ester Hydrolases