{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,4,14]],"date-time":"2026-04-14T06:12:13Z","timestamp":1776147133522,"version":"3.50.1"},"reference-count":52,"publisher":"Frontiers Media SA","license":[{"start":{"date-parts":[[2026,4,14]],"date-time":"2026-04-14T00:00:00Z","timestamp":1776124800000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"content-domain":{"domain":["frontiersin.org"],"crossmark-restriction":true},"short-container-title":["Front. Microbiol."],"abstract":"\n Introduction<\/jats:title>\n \n Control of\n Stegomyia aegypti<\/jats:italic>\n (\n St. aegypti<\/jats:italic>\n ), a major vector of arboviruses, has been increasingly challenged by insecticide resistance, driven by genetic, metabolic, behavioral, and environmental factors. While extensively studied in other contexts, the role of gut microbiota in insecticide resistance remains largely unexplored. Therefore, this study assessed the \u03b1-cypermethrin susceptibility status of\n St. aegypti<\/jats:italic>\n from Puducherry by characterizing the culturable gut bacteria from resistant and susceptible mosquito populations and evaluating their role in mediating resistance.\n <\/jats:p>\n <\/jats:sec>\n \n Materials and methods<\/jats:title>\n \n Mosquito larvae were collected from Puducherry, India, reared, and emerged as female\n St. aegypti<\/jats:italic>\n , which were tested for \u03b1-CP (0.05%) susceptibility following World Health Organisation (WHO) guidelines, along with laboratory strains as controls. Larval and adult guts were dissected, and culturable bacteria were isolated and identified using MALDI-TOF MS and 16S rRNA sequencing. The resistant populations were treated with vancomycin, gentamicin, and streptomycin to suppress the gut microbiota and reassessed for \u03b1-CP susceptibility. The gut bacterial load was quantified by qPCR, and bacterial isolates were evaluated for\n in vitro<\/jats:italic>\n degradation ability in minimal salt medium containing \u03b1-CP.\n <\/jats:p>\n <\/jats:sec>\n \n Results<\/jats:title>\n \n The\n St. aegypti<\/jats:italic>\n populations tested from three locations in Puducherry exhibited confirmed resistance to \u03b1-CP. A total of 35 gut bacterial isolates were obtained (\u03b1-CP-resistant \u221226 and \u03b1-CP-susceptible \u22129). Compared to susceptible mosquitoes, resistant populations showed greater gut bacterial diversity and higher bacterial load. The resistant mosquitoes were dominated by\n Bacillota<\/jats:italic>\n (adults 46.6%, larvae 54.5%) and\n Pseudomonadota<\/jats:italic>\n (adults 46.6%, larvae 45.4%), with\n Bacillaceae<\/jats:italic>\n and\n Enterobacteriaceae<\/jats:italic>\n being the most abundant. The antibiotic-mediated microbiota suppression increased mortality (vancomycin 97%, gentamicin 95%, streptomycin 92%) and reduced LT50 following \u03b1-CP exposure. Further, isolates such as\n Enterobacter hormaechei<\/jats:italic>\n and\n Bacillus<\/jats:italic>\n spp. demonstrated growth in \u03b1-CP supplemented minimal salt media.\n <\/jats:p>\n <\/jats:sec>\n \n Discussion<\/jats:title>\n \n These findings suggest a strong association between gut microbiota and \u03b1-CP resistance in\n St. aegypti.<\/jats:italic>\n The higher microbial diversity, evenness, and bacterial load observed in resistant populations, along with increased mortality following antibiotic treatment and the ability of certain isolates to grow in \u03b1-CP\u2013supplemented media, indicate a potential symbiont-mediated mechanism contributing to insecticide resistance.\n <\/jats:p>\n <\/jats:sec>","DOI":"10.3389\/fmicb.2026.1749347","type":"journal-article","created":{"date-parts":[[2026,4,14]],"date-time":"2026-04-14T05:36:32Z","timestamp":1776144992000},"update-policy":"https:\/\/doi.org\/10.3389\/crossmark-policy","source":"Crossref","is-referenced-by-count":0,"title":["Diversity of culturable gut bacteria and their role in conferring resistance to alpha-cypermethrin in field populations of Stegomyia aegypti"],"prefix":"10.3389","volume":"17","author":[{"given":"Mahima K","family":"Mani","sequence":"first","affiliation":[{"name":"Department of Microbiology and Immunology, ICMR-Vector Control Research Centre","place":["Puducherry, India"]}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Sankari","family":"Thirumal","sequence":"additional","affiliation":[{"name":"Department of Clinical and Molecular Medicine, ICMR-Vector Control Research Centre","place":["Puducherry, India"]}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Vijayakumar","family":"Balakrishnan","sequence":"additional","affiliation":[{"name":"Division of Biostatistics and VBD Modeling, ICMR-Vector Control Research Centre","place":["Puducherry, India"]}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Rakesh","family":"Singh","sequence":"additional","affiliation":[{"name":"Department of Microbiology, Jawaharlal Institute of Postgraduate Medical Education and Research","place":["Puducherry, India"]}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"I","family":"Geetha","sequence":"additional","affiliation":[{"name":"Department of Microbiology and Immunology, ICMR-Vector Control Research Centre","place":["Puducherry, India"]}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"1965","published-online":{"date-parts":[[2026,4,14]]},"reference":[{"key":"ref1","doi-asserted-by":"publisher","first-page":"e40401","DOI":"10.1371\/journal.pone.0040401","article-title":"Serratia odorifera a midgut inhabitant of Aedes aegypti mosquito enhances its susceptibility to Dengue-2 virus","volume":"7","author":"Apte-Deshpande","year":"2012","journal-title":"PLoS One"},{"key":"ref2","doi-asserted-by":"publisher","first-page":"2197","DOI":"10.3390\/v14102197","article-title":"Association of Midgut Bacteria and Their Metabolic Pathways with zika infection and insecticide resistance in Colombian Aedes aegypti populations","volume":"14","author":"Ar\u00e9valo-Cort\u00e9s","year":"2022","journal-title":"Viruses"},{"key":"ref3","doi-asserted-by":"publisher","first-page":"584","DOI":"10.3390\/insects11090584","article-title":"The midgut microbiota of Colombian Aedes aegypti populations with different levels of resistance to the insecticide lambda-cyhalothrin","volume":"11","author":"Ar\u00e9valo-Cort\u00e9s","year":"2020","journal-title":"Insects"},{"key":"ref4","doi-asserted-by":"publisher","first-page":"343","DOI":"10.1099\/00207713-41-3-343","article-title":"Comparative analysis of Bacillus anthracis, Bacillus cereus, and related species on the basis of reverse transcriptase sequencing of 16S rRNA","volume":"41","author":"Ash","year":"1991","journal-title":"Microbiol. Soc."},{"key":"ref5","doi-asserted-by":"publisher","first-page":"1309","DOI":"10.3390\/v15061309","article-title":"Gut bacterial diversity of field and laboratory-reared Aedes albopictus populations of Rio de Janeiro, Brazil","volume":"15","author":"Baltar","year":"2023","journal-title":"Viruses"},{"key":"ref6","doi-asserted-by":"publisher","first-page":"9117","DOI":"10.1038\/s41598-019-45499-z","article-title":"The contribution of gut bacteria to insecticide resistance and the life histories of the major malaria vector Anopheles arabiensis (Diptera: Culicidae)","volume":"9","author":"Barnard","year":"2019","journal-title":"Sci. Rep."},{"key":"ref7","doi-asserted-by":"publisher","first-page":"504","DOI":"10.1038\/nature12060","article-title":"The global distribution and burden of dengue","volume":"496","author":"Bhatt","year":"2013","journal-title":"Nature"},{"key":"ref8","doi-asserted-by":"publisher","first-page":"223","DOI":"10.3390\/microorganisms8020223","article-title":"Enhanced cypermethrin degradation kinetics and metabolic pathway in Bacillus thuringiensis strain SG4","volume":"8","author":"Bhatt","year":"2020","journal-title":"Microorganisms"},{"key":"ref9","doi-asserted-by":"publisher","first-page":"2084","DOI":"10.1038\/s41598-018-20367-4","article-title":"Whole metagenome sequencing reveals links between mosquito microbiota and insecticide resistance in malaria vectors","volume":"8","author":"Dada","year":"2018","journal-title":"Sci. Rep."},{"key":"ref10","doi-asserted-by":"publisher","first-page":"106445","DOI":"10.1016\/j.pestbp.2025.106445","article-title":"A symbiotic bacterium regulates the detoxification metabolism of deltamethrin in Aedes albopictus","volume":"212","author":"Deng","year":"2025","journal-title":"Pestic. Biochem. Physiol."},{"key":"ref11","doi-asserted-by":"publisher","first-page":"384","DOI":"10.1186\/s12866-025-04114-0","article-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","volume":"25","author":"Djondji Kamga","year":"2025","journal-title":"BMC Microbiol."},{"key":"ref12","doi-asserted-by":"publisher","first-page":"9047","DOI":"10.1073\/pnas.0610451104","article-title":"Bacteria of the genus Asaia stably associate with Anopheles stephensi, an Asian malarial mosquito vector | PNAS","volume":"104","author":"Favia","year":"2007","journal-title":"Proc. Natl. Acad. Sci."},{"key":"ref13","doi-asserted-by":"publisher","first-page":"105174","DOI":"10.1016\/j.actatropica.2019.105174","article-title":"Use of MALDI-TOF MS to identify the culturable midgut microbiota of laboratory and wild mosquitoes","volume":"200","author":"Gazzoni Ara\u00fajo Gon\u00e7alves","year":"2019","journal-title":"Acta Trop."},{"key":"ref14","doi-asserted-by":"publisher","first-page":"715","DOI":"10.1016\/j.meegid.2014.09.029","article-title":"Composition of Anopheles coluzzii and Anopheles gambiae microbiota from larval to adult stages","volume":"28","author":"Gimonneau","year":"2014","journal-title":"Infect. Genet. Evol."},{"key":"ref15","doi-asserted-by":"publisher","first-page":"761459","DOI":"10.3389\/fmicb.2022.761459","article-title":"Suppression of midgut microbiota impact pyrethroid susceptibility in Aedes aegypti","volume":"13","author":"G\u00f3mez-Govea","year":"2022","journal-title":"Front. Microbiol."},{"key":"ref16","doi-asserted-by":"publisher","first-page":"49","DOI":"10.1186\/s40168-018-0435-2","article-title":"The mosquito holobiont: fresh insight into mosquito-microbiota interactions","volume":"6","author":"Gu\u00e9gan","year":"2018","journal-title":"Microbiome"},{"key":"ref17","doi-asserted-by":"publisher","first-page":"1154","DOI":"10.1038\/s42003-023-05545-z","article-title":"The mosquito Aedes aegypti requires a gut microbiota for normal fecundity, longevity and vector competence","volume":"6","author":"Harrison","year":"2023","journal-title":"Commun. Biol."},{"key":"ref18","doi-asserted-by":"publisher","first-page":"20130371","DOI":"10.1098\/rspb.2013.0371","article-title":"Facilitating Wolbachia introductions into mosquito populations through insecticide-resistance selection | proceedings of the Royal Society B: biological sciences","volume":"280","author":"Hoffmann","year":"2013","journal-title":"Proc. R. Soc. B Biol. Sci."},{"key":"ref19","doi-asserted-by":"publisher","first-page":"1019","DOI":"10.1007\/s42690-021-00670-w","article-title":"Spatial distribution of insecticide resistance and susceptibility in Aedes aegypti and Aedes albopictus in India","volume":"42","author":"Jangir","year":"2022","journal-title":"Int. J. Trop. Insect Sci."},{"key":"ref20","first-page":"952","article-title":"Isolation and characterization of comprehensive polychlorinated biphenyl degrading bacterium, Enterobacter sp. LY402","volume":"18","author":"Jia","year":"2008","journal-title":"J. Microbiol. Biotechnol."},{"key":"ref21","doi-asserted-by":"publisher","first-page":"220","DOI":"10.1111\/eva.12532","article-title":"Rapid evolution of symbiont-mediated resistance compromises biological control of aphids by parasitoids","volume":"11","author":"K\u00e4ch","year":"2018","journal-title":"Evol. Appl."},{"key":"ref22","doi-asserted-by":"publisher","first-page":"8618","DOI":"10.1073\/pnas.1200231109","article-title":"Symbiont-mediated insecticide resistance","volume":"109","author":"Kikuchi","year":"2012","journal-title":"Proc. Natl. Acad. Sci. USA"},{"key":"ref23","doi-asserted-by":"publisher","first-page":"346","DOI":"10.1099\/ijs.0.059774-0","article-title":"Towards a taxonomic coherence between average nucleotide identity and 16S rRNA gene sequence similarity for species demarcation of prokaryotes","volume":"64","author":"Kim","year":"2014","journal-title":"Int. J. Syst. Evol. Microbiol."},{"key":"ref24","doi-asserted-by":"publisher","first-page":"1644","DOI":"10.1126\/science.1184008","article-title":"A peroxidase\/dual oxidase system modulates midgut epithelial immunity in Anopheles gambiae","volume":"327","author":"Kumar","year":"2010","journal-title":"Science (N. Y.)"},{"key":"ref25","first-page":"2445","article-title":"Isolation and identification of degradation bacteria Enterobacter aerogenes for pyrethriods pesticide residues and its degradation characteristics","volume":"30","author":"Liao","year":"2009","journal-title":"Huan Jing Ke Xue = Huanjing Kexue"},{"key":"ref26","doi-asserted-by":"publisher","first-page":"650743","DOI":"10.3389\/fmicb.2021.650743","article-title":"Diet\u2013microbiota interactions alter mosquito development","volume":"12","author":"Martinson","year":"2021","journal-title":"Front. Microbiol."},{"key":"ref27","doi-asserted-by":"publisher","first-page":"146","DOI":"10.1186\/1756-3305-6-146","article-title":"Diversity and function of bacterial microbiota in the mosquito holobiont","volume":"6","author":"Minard","year":"2013","journal-title":"Parasit. Vectors"},{"key":"ref28","doi-asserted-by":"publisher","first-page":"29","DOI":"10.1603\/0022-2585-38.1.29","article-title":"Meconial peritrophic membranes and the fate of midgut bacteria during mosquito (Diptera: Culicidae) metamorphosis","volume":"38","author":"Moll","year":"2001","journal-title":"J. Med. Entomol."},{"key":"ref29","doi-asserted-by":"publisher","first-page":"117","DOI":"10.1080\/09603123.2018.1515425","article-title":"Full article: prevalence and diversity of blaTEM, blaSHV and blaCTX-M variants among multidrug resistant Klebsiella spp. from an urban riverine environment in India","volume":"29","author":"Mondal","year":"2019","journal-title":"Int. J. Environ. Health Res."},{"key":"ref30","doi-asserted-by":"publisher","first-page":"14406","DOI":"10.1038\/s41598-021-93725-4","article-title":"Resistance to permethrin alters the gut microbiota of Aedes aegypti","volume":"11","author":"Muturi","year":"2021","journal-title":"Sci. Rep."},{"key":"ref31","doi-asserted-by":"publisher","first-page":"e1001320","DOI":"10.1371\/journal.ppat.1001320","article-title":"Blood meal-derived heme decreases ROS levels in the midgut of Aedes aegypti and allows proliferation of intestinal microbiota","volume":"7","author":"Oliveira","year":"2011","journal-title":"PLoS Pathog."},{"key":"ref32","doi-asserted-by":"publisher","first-page":"e1007725","DOI":"10.1371\/journal.pgen.1007725","article-title":"A distinct strain of Arsenophonus symbiont decreases insecticide resistance in its insect host","volume":"14","author":"Pang","year":"2018","journal-title":"PLoS Genet."},{"key":"ref33","doi-asserted-by":"publisher","first-page":"14","DOI":"10.1093\/jisesa\/iead019","article-title":"Isolation and characterization of gut bacteria associated with the degradation of host-specific terpenoids in Pagiophloeus tsushimanus (Coleoptera: Curculionidae) larvae","volume":"23","author":"Qiao","year":"2023","journal-title":"J. Insect Sci."},{"key":"ref34","doi-asserted-by":"publisher","first-page":"1162","DOI":"10.1002\/mbo3.1026","article-title":"Variation in the microbiota across different developmental stages of Aedes albopictus is affected by ampicillin exposure","volume":"9","author":"Qing","year":"2020","journal-title":"Microbiolopen"},{"key":"ref35","doi-asserted-by":"publisher","first-page":"792","DOI":"10.1089\/mdr.2017.0078","article-title":"Prevalence and molecular characterization of New Delhi Metallo-Beta-lactamases in multidrug-resistant Pseudomonas aeruginosa and Acinetobacter baumannii from India","volume":"24","author":"Rahman","year":"2018","journal-title":"Microb. Drug Resist."},{"key":"ref36","author":"Rueda","year":"2004"},{"key":"ref37","doi-asserted-by":"publisher","first-page":"947857","DOI":"10.3389\/fmicb.2022.947857","article-title":"Studies on the fitness characteristics of wMel- and wAlbB-introgressed Aedes aegypti (Pud) lines in comparison with wMel- and wAlbB-transinfected Aedes aegypti (Aus) and wild-type Aedes aegypti (Pud) lines","volume":"13","author":"Sadanandane","year":"2022","journal-title":"Front. Microbiol."},{"key":"ref38","doi-asserted-by":"publisher","first-page":"e0005677","DOI":"10.1371\/journal.pntd.0005677","article-title":"Amino acid metabolic signaling influences Aedes aegypti midgut microbiome variability","volume":"11","author":"Short","year":"2017","journal-title":"PLoS Negl. Trop. Dis."},{"key":"ref39","doi-asserted-by":"publisher","first-page":"4855","DOI":"10.1128\/AEM.70.8.4855-4863.2004","article-title":"Biodegradation of chlorpyrifos by Enterobacter strain B-14 and its use in bioremediation of contaminated soils","volume":"70","author":"Singh","year":"2004","journal-title":"Appl. Environ. Microbiol."},{"key":"ref40","doi-asserted-by":"publisher","first-page":"289","DOI":"10.1080\/20477724.2017.1356052","article-title":"The role of midgut symbiotic bacteria in resistance of Anopheles stephensi (Diptera: Culicidae) to organophosphate insecticides","volume":"111","author":"Soltani","year":"2017","journal-title":"Pathog. Glob. Health"},{"key":"ref41","doi-asserted-by":"publisher","first-page":"3022","DOI":"10.1093\/molbev\/msab120","article-title":"MEGA11: molecular evolutionary genetics analysis version 11","volume":"38","author":"Tamura","year":"2021","journal-title":"Mol. Biol. Evol."},{"key":"ref42","doi-asserted-by":"publisher","first-page":"1170","DOI":"10.1017\/S0031182018000070","article-title":"Using MALDI-TOF MS to identify mosquitoes collected in Mali and their blood meals","volume":"145","author":"Tandina","year":"2018","journal-title":"Parasitology"},{"key":"ref43","doi-asserted-by":"publisher","first-page":"181","DOI":"10.3390\/insects16020181","article-title":"Diversity of gut bacteria of field-collected Aedes aegypti larvae and females, resistant to temephos and deltamethrin","volume":"16","author":"Viafara-Campo","year":"2025","journal-title":"Insects"},{"key":"ref44","doi-asserted-by":"publisher","first-page":"e0010208","DOI":"10.1371\/journal.pntd.0010208","article-title":"A symbiotic gut bacterium enhances Aedes albopictus resistance to insecticide","volume":"16","author":"Wang","year":"2022","journal-title":"PLoS Negl. Trop. Dis."},{"key":"ref45","doi-asserted-by":"publisher","first-page":"105577","DOI":"10.1016\/j.pestbp.2023.105577","article-title":"Insecticide resistance: status and potential mechanisms in Aedes aegypti","volume":"195","author":"Wang","year":"2023","journal-title":"Pestic. Biochem. Physiol."},{"key":"ref46","doi-asserted-by":"publisher","first-page":"e1177","DOI":"10.1002\/mbo3.1177","article-title":"Differences in the intestinal microbiota between insecticide-resistant and-sensitive Aedes albopictus based on full-length 16S rRNA sequencing","volume":"10","author":"Wang","year":"2021","journal-title":"Microbiology"},{"key":"ref47","doi-asserted-by":"publisher","first-page":"5994","DOI":"10.1073\/pnas.1703546114","article-title":"Insect pathogenic fungus interacts with the gut microbiota to accelerate mosquito mortality","volume":"114","author":"Wei","year":"2017","journal-title":"Proc. Natl. Acad. Sci."},{"key":"ref48","doi-asserted-by":"publisher","first-page":"697","DOI":"10.1128\/jb.173.2.697-703.1991","article-title":"16S ribosomal DNA amplification for phylogenetic study | journal of bacteriology","volume":"173","author":"Weisburg","year":"1991","journal-title":"J. Bacteriol."},{"key":"ref49","doi-asserted-by":"publisher","first-page":"17","DOI":"10.1186\/s13071-024-06638-2","article-title":"Insecticide resistant Anopheles from Ethiopia but not Burkina Faso show a microbiota composition shift upon insecticide exposure","volume":"18","author":"Worku","year":"2025","journal-title":"Parasites Vectors"},{"key":"ref50","article-title":"Standard operating procedure for testing insecticide susceptibility of adult mosquitoes in WHO tube tests","volume-title":"Standard Operating Procedure for Testing Insecticide Susceptibility of Adult Mosquitoes in WHO Tube Tests","year":"2022"},{"key":"ref51","doi-asserted-by":"publisher","first-page":"e68852","DOI":"10.1371\/journal.pone.0068852","article-title":"DNA sequencing reveals the midgut microbiota of diamondback moth, Plutella xylostella (L.) and a possible relationship with insecticide resistance","volume":"8","author":"Xia","year":"2013","journal-title":"PLoS One"},{"key":"ref52","doi-asserted-by":"publisher","first-page":"2849","DOI":"10.1007\/s00253-014-6164-y","article-title":"Isolation of a novel beta-cypermethrin degrading strain Bacillus subtilis BSF01 and its biodegradation pathway |","volume":"99","author":"Xiao","year":"2015","journal-title":"Appl. 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