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Assessing the extent of this aggravation and its impact on the efficacy of these tools is vital to ensure the continued control of major vectors. Here we took advantage of 2009 and 2014 data from Malawi to establish the extent of the resistance escalation in 2021 and assessed its impact on various bed nets performance.<\/jats:p><\/jats:sec><jats:sec><jats:title>Methods<\/jats:title><jats:p>Indoor blood-fed and wild female Anopheles (An) mosquitoes were collected with an electric aspirator in Chikwawa. Cocktail and SINE PCR were used to identify sibling species belonging to<jats:italic>An. funestus<\/jats:italic>group and<jats:italic>An. gambiae<\/jats:italic>complex. The susceptibility profile to the four classes of insecticides was assessed using the WHO tubes bioassays. Data were saved in an Excel file. Analysis was done using Vassarstats and figures by Graph Pad.<\/jats:p><\/jats:sec><jats:sec><jats:title>Results<\/jats:title><jats:p>In this study, a high level of resistance was observed with pyrethroids (permethrin, deltamethrin and alpha-cypermethrin with mortality rate at 5x discriminating concentration (DC)\u2009&lt;\u200950% and Mortality rate at 10x DC\u2009&lt;\u200970%). A high level of resistance was also observed to carbamate (bendiocarb) with mortality rate at 5x DC\u2009&lt;\u200925%). Aggravation of resistance was also noticed between 2009 and 2021. For pyrethroids, the mortality rate for permethrin reduced from 47.2% in 2009 to 13% in 2014 and 6.7% in 2021. For deltamethrin, the mortality rate reduced from 42.3% in 2009 to 1.75% in 2014 and 5.2% in 2021. For Bendiocarb, the mortality rate reduced from 60% in 2009 to 30.1% in 2014 and 12.2% in 2021. The high resistance observed is consistent with a drastic loss of pyrethroid-only bed nets efficacy although Piperonyl butoxide (PBO)-based nets remain effective. The resistance pattern observed was linked with high up-regulation of the P450 genes<jats:italic>CYP6P9a, CYP6P9b<\/jats:italic>and<jats:italic>CYP6M7<\/jats:italic>in<jats:italic>An. funestus<\/jats:italic>s.s. mosquitoes surviving exposure to deltamethrin at 1x, 5x and 10x DC. A significant association was observed between the 6.5\u00a0kb structural variant and resistance escalation with homozygote resistant (SV+\/SV+) more likely to survive exposure to 5x and 10x (OR\u2009=\u20094.1; P\u2009&lt;\u20090.001) deltamethrin than heterozygotes. However, a significant proportion of mosquitoes survived the synergist assays with PBO suggesting that other mechanisms than P450s are present.<\/jats:p><\/jats:sec><jats:sec><jats:title>Conclusions<\/jats:title><jats:p>This resistance aggravation in<jats:italic>An. funestus<\/jats:italic>s.s. Malawian population highlights an urgent need to deploy novel control tools not relying on pyrethroids to improve the effectiveness of vector control.<\/jats:p><\/jats:sec>","DOI":"10.1186\/s12879-022-07596-9","type":"journal-article","created":{"date-parts":[[2022,7,30]],"date-time":"2022-07-30T05:03:53Z","timestamp":1659157433000},"update-policy":"https:\/\/doi.org\/10.1007\/springer_crossmark_policy","source":"Crossref","is-referenced-by-count":39,"title":["Marked aggravation of pyrethroid resistance in major malaria vectors in Malawi between 2014 and 2021 is partly linked with increased expression of P450 alleles"],"prefix":"10.1186","volume":"22","author":[{"given":"Benjamin D.","family":"Menze","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Magellan","family":"Tchouakui","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Leon M. 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