Metabolomic Unravelling and RSM-Assisted Optimization of Phenylpyrazole Degradation by a Novel Soil-Derived Bacterial Consortium FP-25

Metabolomic Unravelling and RSM-Assisted Optimization of Phenylpyrazole Degradation by a Novel Soil-Derived Bacterial Consortium FP-25 | Research Square window.SnipcartSettings = { analytics: { enabled: false } }; (function() { var accessVector = localStorage.getItem('access_vector') || ''; window.dataLayer = window.dataLayer || []; if (accessVector) { window.dataLayer.push({ user: { profile: { profileInfo: { snid: accessVector } } } }); } })(); (function(w,d,s,l,i){w[l]=w[l]||[];w[l].push({'gtm.start':new Date().getTime(),event:'gtm.js'});var f=d.getElementsByTagName(s)[0],j=d.createElement(s),dl=l!='dataLayer'?'&l='+l:'';j.async=true;j.src='https://www.googletagmanager.com/gtm.js?id='+i+dl;f.parentNode.insertBefore(j,f);})(window,document,'script','dataLayer','GTM-K279D39R'); Browse Preprints In Review Journals COVID-19 Preprints AJE Video Bytes Research Tools Research Promotion AJE Professional Editing AJE Rubriq About Preprint Platform In Review Editorial Policies Our Team Advisory Board Help Center Sign In Submit a Preprint Cite Share Download PDF Research Article Metabolomic Unravelling and RSM-Assisted Optimization of Phenylpyrazole Degradation by a Novel Soil-Derived Bacterial Consortium FP-25 Shivani Uniyal, Rashmi Paliwal, Rohit Mahar, J.P.N. Rai This is a preprint; it has not been peer reviewed by a journal. https://doi.org/ 10.21203/rs.3.rs-7764059/v1 This work is licensed under a CC BY 4.0 License Status: Posted Version 1 posted You are reading this latest preprint version Abstract Fipronil is a phenylpyrazole insecticide that is frequently utilized in agricultural practices. It engenders noteworthy environmental threat owing to its persistence, bioaccumulation, and off-target toxicity. The empirical bioremediation propensity of known fipronil-degrading bacteria is restrained by its inferior efficiency and low environmental resilience. In this study, a novel microbial consortium, FP-25, was concocted by isolating two fipronil-degrading indigenous bacterial strains— Pseudomonas furukawaii strain S4 and Agrobacterium pusense strain S6—from perpetually contaminated agricultural soil. The consortium was appraised for its fipronil degradation propensity in aqueous media. Optimization of biodegradation variables was orchestrated through response surface methodology (RSM) deploying a Box–Behnken design, which methodically analyzed the interactive effects of pH, temperature, inoculum biomass and fipronil concentration. The optimized conditions—32.5°C, neutral pH, and an inoculum concentration of 0.175 g L⁻¹, and 200 mg L − 1 fipronil concentration —effectuated degradation efficiencies of 91.92% for consortium FP-25 within 14 days of incubation. The generation of degradation products was certainly corroborated through GC-MS analysis. Consequently, a catabolic pathway for fipronil degradation used by the consortium FP-25 has been explicated, depicting the successive enzymatic transformation of fipronil to non-toxic metabolites. The befitting and the rationality of the RSM model were evaluated through the deployment of an in-situ microcosm experiment, utilizing actual contaminated soil sampled from the Himalayan highland ecosystem. Degradation kinetics substantiated first-order reaction models with rate constants ranging between 0.046 and 0.076 day⁻¹. Thus, the present study endorsed the puissant bioremediation ability of the developed consortium FP-25 as an eco-friendly and sustainable substitute for conventional approaches. Fipronil Bioremediation Pseudomonas furukawaii Agrobacterium pusense Gas chromatography Full Text Supplementary Files Supplementrymaterial3.docx Cite Share Download PDF Status: Posted Version 1 posted You are reading this latest preprint version Research Square lets you share your work early, gain feedback from the community, and start making changes to your manuscript prior to peer review in a journal. As a division of Research Square Company, we’re committed to making research communication faster, fairer, and more useful. We do this by developing innovative software and high quality services for the global research community. Our growing team is made up of researchers and industry professionals working together to solve the most critical problems facing scientific publishing. Also discoverable on Platform About Our Team In Review Editorial Policies Advisory Board Help Center Resources Author Services Accessibility API Access RSS feed Manage Cookie Preferences © Research Square 2026 | ISSN 2693-5015 (online) Privacy Policy Terms of Service Do Not Sell My Personal Information {"props":{"pageProps":{"initialData":{"identity":"rs-7764059","acceptedTermsAndConditions":true,"allowDirectSubmit":true,"archivedVersions":[],"articleType":"Research Article","associatedPublications":[],"authors":[{"id":525722326,"identity":"faa8aee4-24da-4b39-b16b-ab11728f71e1","order_by":0,"name":"Shivani Uniyal","email":"data:image/png;base64,iVBORw0KGgoAAAANSUhEUgAAAZAAAAAyAQMAAABI0h/eAAAABlBMVEX///8AAABVwtN+AAAACXBIWXMAAA7EAAAOxAGVKw4bAAABFUlEQVRIiWNgGAWjYDACCcYGEMnAwAykHjYwJPCDRBMKiNWSCNQiCeInGODTgswBaTE4AGLh0SI/u7lN4ucOi8Tt7DyGHxJ33MkzPr868cMDAwZ5frEDWLUY3DnYJtl7RiJxZzOPsUTimWfFZjfebpYAOsxw5uwE7FokEpsNeNskEjcc5gGy2w4nbrtxdgNIS4LBbexa5GckNhv+hWgx/gHSsnnG2c0/8GlhuJHY+BhqixnYlg38vdvw2mIA0iLbJmG84TBbmQVQS7HEDd5tFgkGEjj9Ij8j/cHBt211shvOH95842Pb4Tz+/rObb/6osJHnl8bhMATggMaFBFilBB6VcMD+AELzHyBG9SgYBaNgFIwgAACi/GmPZ16VcgAAAABJRU5ErkJggg==","orcid":"","institution":"Hemwati Nandan Bahuguna Garhwal University - Swami Ram Teerth Campus Badshahi Thaul Tehri","correspondingAuthor":true,"prefix":"","firstName":"Shivani","middleName":"","lastName":"Uniyal","suffix":""},{"id":525722327,"identity":"b4115697-9312-43db-8c78-76fabfe0a768","order_by":1,"name":"Rashmi Paliwal","email":"","orcid":"","institution":"Kurukshetra University","correspondingAuthor":false,"prefix":"","firstName":"Rashmi","middleName":"","lastName":"Paliwal","suffix":""},{"id":525722328,"identity":"3aebe452-d433-4b31-8b1c-d00bbb7fd06c","order_by":2,"name":"Rohit Mahar","email":"","orcid":"","institution":"Hemvati Nandan Bahuguna Garhwal University","correspondingAuthor":false,"prefix":"","firstName":"Rohit","middleName":"","lastName":"Mahar","suffix":""},{"id":525722329,"identity":"a128b74a-4152-4fe0-823a-bf8a0d12694a","order_by":3,"name":"J.P.N. 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FP-25","fulltext":[],"fulltextSource":"","fullText":"","funders":[],"hasAdminPriorityOnWorkflow":false,"hasManuscriptDocX":false,"hasOptedInToPreprint":true,"hasPassedJournalQc":"","hasAnyPriority":false,"hideJournal":true,"highlight":"","institution":"","isAcceptedByJournal":false,"isAuthorSuppliedPdf":true,"isDeskRejected":"","isHiddenFromSearch":false,"isInQc":false,"isInWorkflow":true,"isPdf":true,"isPdfUpToDate":true,"isWithdrawnOrRetracted":false,"journal":{"display":true,"email":"[email protected]","identity":"researchsquare","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":true,"externalIdentity":"","sideBox":"","snPcode":"","submissionUrl":"/submission","title":"Research Square","twitterHandle":"researchsquare","acdcEnabled":true,"dfaEnabled":false,"editorialSystem":"","reportingPortfolio":"","inReviewEnabled":false,"inReviewRevisionsEnabled":true},"keywords":"Fipronil, Bioremediation, Pseudomonas furukawaii, Agrobacterium pusense, Gas chromatography","lastPublishedDoi":"10.21203/rs.3.rs-7764059/v1","lastPublishedDoiUrl":"https://doi.org/10.21203/rs.3.rs-7764059/v1","license":{"name":"CC BY 4.0","url":"https://creativecommons.org/licenses/by/4.0/"},"manuscriptAbstract":"\u003cp\u003eFipronil is a phenylpyrazole insecticide that is frequently utilized in agricultural practices. It engenders noteworthy environmental threat owing to its persistence, bioaccumulation, and off-target toxicity. The empirical bioremediation propensity of known fipronil-degrading bacteria is restrained by its inferior efficiency and low environmental resilience. In this study, a novel microbial consortium, FP-25, was concocted by isolating two fipronil-degrading indigenous bacterial strains\u0026mdash;\u003cem\u003ePseudomonas furukawaii\u003c/em\u003e strain S4 and \u003cem\u003eAgrobacterium pusense\u003c/em\u003e strain S6\u0026mdash;from perpetually contaminated agricultural soil. The consortium was appraised for its fipronil degradation propensity in aqueous media. Optimization of biodegradation variables was orchestrated through response surface methodology (RSM) deploying a Box\u0026ndash;Behnken design, which methodically analyzed the interactive effects of pH, temperature, inoculum biomass and fipronil concentration. The optimized conditions\u0026mdash;32.5\u0026deg;C, neutral pH, and an inoculum concentration of 0.175 g L⁻\u0026sup1;, and 200 mg L\u003csup\u003e\u0026minus;\u0026thinsp;1\u003c/sup\u003e fipronil concentration \u0026mdash;effectuated degradation efficiencies of 91.92% for consortium FP-25 within 14 days of incubation. The generation of degradation products was certainly corroborated through GC-MS analysis. Consequently, a catabolic pathway for fipronil degradation used by the consortium FP-25 has been explicated, depicting the successive enzymatic transformation of fipronil to non-toxic metabolites. The befitting and the rationality of the RSM model were evaluated through the deployment of an in-situ microcosm experiment, utilizing actual contaminated soil sampled from the Himalayan highland ecosystem. Degradation kinetics substantiated first-order reaction models with rate constants ranging between 0.046 and 0.076 day⁻\u0026sup1;. Thus, the present study endorsed the puissant bioremediation ability of the developed consortium FP-25 as an eco-friendly and sustainable substitute for conventional approaches.\u003c/p\u003e","manuscriptTitle":"Metabolomic Unravelling and RSM-Assisted Optimization of Phenylpyrazole Degradation by a Novel Soil-Derived Bacterial Consortium FP-25","msid":"","msnumber":"","nonDraftVersions":[{"code":1,"date":"2025-10-22 14:45:52","doi":"10.21203/rs.3.rs-7764059/v1","editorialEvents":[{"type":"communityComments","content":0}],"status":"published","journal":{"display":true,"email":"[email protected]","identity":"researchsquare","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":true,"externalIdentity":"","sideBox":"","snPcode":"","submissionUrl":"/submission","title":"Research Square","twitterHandle":"researchsquare","acdcEnabled":true,"dfaEnabled":false,"editorialSystem":"","reportingPortfolio":"","inReviewEnabled":false,"inReviewRevisionsEnabled":true}}],"origin":"","ownerIdentity":"53973a56-1d59-405f-9df3-bc421ca4a51e","owner":[],"postedDate":"October 22nd, 2025","published":true,"recentEditorialEvents":[],"rejectedJournal":[],"revision":"","amendment":"","status":"posted","subjectAreas":[],"tags":[],"updatedAt":"2026-03-01T03:50:35+00:00","versionOfRecord":[],"versionCreatedAt":"2025-10-22 14:45:52","video":"","vorDoi":"","vorDoiUrl":"","workflowStages":[]},"version":"v1","identity":"rs-7764059","journalConfig":"researchsquare"},"__N_SSP":true},"page":"/article/[identity]/[[...version]]","query":{"redirect":"/article/rs-7764059","identity":"rs-7764059","version":["v1"]},"buildId":"8U1c8b4HqxoKbykW_rLl7","isFallback":false,"isExperimentalCompile":false,"dynamicIds":[84888],"gssp":true,"scriptLoader":[]}