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However, the mechanisms enabling the vectors to now survive exposure to very high doses of pyrethroids remain unclear. Here, using High-throughput sequencing of the 16\u00a0S ribosomal RNA gene coupled with antibiotic treatment, we provide evidence linking the mosquito microbiome to the escalation of pyrethroid resistance in major African malaria vectors,\n                      <jats:italic>Anopheles gambiae<\/jats:italic>\n                      (s.s.) and\n                      <jats:italic>Anopheles funestus<\/jats:italic>\n                      (s.s.).\n                    <\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec>\n                    <jats:title>Results<\/jats:title>\n                    <jats:p>\n                      Phenotypic characterisation of\n                      <jats:italic>An. gambiae<\/jats:italic>\n                      (s.s.) and\n                      <jats:italic>An. funestus<\/jats:italic>\n                      (s.s.) populations revealed a high level of resistance to pyrethroid in both species, with mortality rates\u2009&lt;\u200991% at 10x the diagnostic dose of each insecticide. A significant difference in bacterial composition was observed in\n                      <jats:italic>An. gambiae<\/jats:italic>\n                      s.s. between resistant mosquitoes exposed to 1X and 10X the diagnostic dose of permethrin, and the susceptible strains (PERMANOVA-F: 8.06;\n                      <jats:italic>p<\/jats:italic>\n                      \u2009=\u20090.02). The abundance of\n                      <jats:italic>Pseudomonas_1<\/jats:italic>\n                      (Log2FC: 4.42,\n                      <jats:italic>p<\/jats:italic>\n                      \u2009=\u20090.0001) and\n                      <jats:italic>Burkholderia_1<\/jats:italic>\n                      (Log2FC: 4.95,\n                      <jats:italic>p<\/jats:italic>\n                      \u2009=\u20090.001) bacteria were consistently associated with mosquitoes surviving 1X and 10X the diagnostic concentrations of permethrin, respectively, while\n                      <jats:italic>Serratia_2<\/jats:italic>\n                      bacteria was mostly associated with insecticide susceptibility. In the\n                      <jats:italic>An. funestus<\/jats:italic>\n                      s.s. strain, there was no significant difference in bacterial alpha- and beta-diversity between the FUMOZ-R (exhibiting normal deltamethrin resistance) and FUMOZ-HR (selected for high deltamethrin resistance), suggesting a minimal impact of selection pressure on bacterial composition. However, in FUMOZ-HR, there was an increase in the abundance of\n                      <jats:italic>Rahnella<\/jats:italic>\n                      (Log2FC: 15.954,\n                      <jats:italic>p<\/jats:italic>\n                      \u2009=\u20099.73 E-12) and\n                      <jats:italic>Leucobacter<\/jats:italic>\n                      (Log2FC: 7.6,\n                      <jats:italic>p<\/jats:italic>\n                      \u2009=\u20090.008) bacteria, indicating their potential role in worsening deltamethrin resistance. Furthermore, treating resistant mosquitoes (both\n                      <jats:italic>Anopheles<\/jats:italic>\n                      species) with broad-spectrum bactericidal antibiotics (penicillin\/streptomycin) via sugar solution increased their susceptibility to various diagnostic doses of permethrin and deltamethrin in WHO pyrethroid intensity bioassays.\n                    <\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec>\n                    <jats:title>Conclusion<\/jats:title>\n                    <jats:p>\n                      Overall, our study emphasises the potential role of the microbiome in the escalation of insecticide resistance in\n                      <jats:italic>Anopheles<\/jats:italic>\n                      mosquitoes, identifying key bacterial strains associated with insecticide resistance and susceptibility. These candidate bacteria warrant further investigation to elucidate the mechanisms by which they contribute to the escalation of pyrethroid resistance.\n                    <\/jats:p>\n                  <\/jats:sec>","DOI":"10.1186\/s12866-025-04114-0","type":"journal-article","created":{"date-parts":[[2025,7,1]],"date-time":"2025-07-01T23:51:07Z","timestamp":1751413867000},"update-policy":"https:\/\/doi.org\/10.1007\/springer_crossmark_policy","source":"Crossref","is-referenced-by-count":10,"title":["Evidence of microbiome contribution to the escalation of pyrethroid resistance in the major malaria vectors Anopheles gambiae s.s. and Anopheles funestus s.s"],"prefix":"10.1186","volume":"25","author":[{"given":"Fleuriane Metissa","family":"Djondji Kamga","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Leon M.","family":"Jean Mugenzi","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Vanessa Brigitte","family":"Ngannang-Fezeu","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Fran\u00e7ois Sougal","family":"Ngambia Freitas","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Calmes Ursain","family":"Bouaka Tsakeng","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Maurice Marcel","family":"Sandeu","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Magellan","family":"Tchouakui","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Charles Sinclair","family":"Wondji","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"297","published-online":{"date-parts":[[2025,7,2]]},"reference":[{"key":"4114_CR1","unstructured":"World malaria report. 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