Dissecting the Phenotypic Complexity of Biofilm Development and Quorum Sensing in Salmonella enterica Serovars

Dissecting the Phenotypic Complexity of Biofilm Development and Quorum Sensing in Salmonella enterica Serovars | 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 Dissecting the Phenotypic Complexity of Biofilm Development and Quorum Sensing in Salmonella enterica Serovars Anjali Ghosh, Schneider Fernandes This is a preprint; it has not been peer reviewed by a journal. https://doi.org/ 10.21203/rs.3.rs-9130366/v1 This work is licensed under a CC BY 4.0 License Status: Under Revision Version 1 posted 17 You are reading this latest preprint version Abstract This study investigates the mechanisms of biofilm formation and quorum sensing in Salmonella enterica serovars, with particular emphasis on their implications for clinical safety and public health. The ability of this bacterium to form biofilms on various surfaces poses significant challenges for control and eradication efforts, contributing to persistent infections and enhanced antimicrobial resistance. The study explores quorum quenching strategies to inhibit biofilm formation through certain treatments. The research employes crystal violet staining, bacterial growth curve analysis, and fluorescence microscopy to evaluate the effects of quorum sensing (QS) and natural product-based quorum quenching strategies on biofilm formation on abiotic (polystyrene) surfaces. Short- to medium-chain acyl-homoserine lactones (AHLs), particularly C8-HSL were assessed for their role in promoting biofilm development, alongside plant-derived and dietary natural products with reported antimicrobial activity, including aloe emodin (from Aloe vera ), epigallocatechin gallate (EGCG) (from green tea, Camellia sinensis ), thymoquinone from ( Nigella sativa) garlic extract (active compound Allium sativum ), and turmeric extract (active extract Curcuma longa ). Results demonstrated that C8-HSL significantly enhanced biofilm formation (Figs. 2–4; Tables 1–2), with 0.2 mM identified as the most effective concentration (Fig. 4). In contrast, the tested natural products exhibited quorum quenching and anti-biofilm activity, significantly reducing biofilm biomass and structural integrity (Figs. 5 and 6). Fluorescence microscopy further supported these findings by demonstrating enhanced biofilm formation in the presence of quorum sensing molecules and disrupted biofilm architecture following quorum quenching treatment (Figs. 7 and 8). These findings highlight the central role of quorum sensing in Salmonella enterica pathogenicity and demonstrate the potential of plant-derived and dietary natural products as alternative quorum quenching agents for biofilm control. The study suggests that further research, including the use of genetically engineered mutants, is required to elucidate the specific genetic pathways involved in QS-mediated biofilm regulation. Overall, this research advances understanding of biofilm formation and quorum sensing in Salmonella enterica and supports the development of novel strategies to manage persistent infections and improve food safety. N-acyl homoserine lactone (AHL) Salmonella Typhimurium 14028 Salmonella Typhimurium Wildtype and Salmonella Enteritidis UCH Salmonella Virchow C8HSL Farnesol CV assay fluorescence microscopy antibiotic susceptibility test Biofilm formation quorum sensing quorum quenching and natural compounds Full Text Additional Declarations No competing interests reported. Cite Share Download PDF Status: Under Revision Version 1 posted Editorial decision: Revision requested 30 Apr, 2026 Reviews received at journal 27 Apr, 2026 Reviews received at journal 26 Apr, 2026 Reviews received at journal 24 Apr, 2026 Reviews received at journal 22 Apr, 2026 Reviews received at journal 18 Apr, 2026 Reviewers agreed at journal 12 Apr, 2026 Reviewers agreed at journal 09 Apr, 2026 Reviewers agreed at journal 09 Apr, 2026 Reviewers agreed at journal 08 Apr, 2026 Reviewers agreed at journal 08 Apr, 2026 Reviewers agreed at journal 08 Apr, 2026 Reviewers agreed at journal 08 Apr, 2026 Reviewers invited by journal 08 Apr, 2026 Editor assigned by journal 07 Apr, 2026 Submission checks completed at journal 07 Apr, 2026 First submitted to journal 06 Apr, 2026 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. 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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-9130366","acceptedTermsAndConditions":true,"allowDirectSubmit":false,"archivedVersions":[],"articleType":"Research Article","associatedPublications":[],"authors":[{"id":621719824,"identity":"d5e6ddd2-41ad-4180-9df9-cecb2076b7ce","order_by":0,"name":"Anjali Ghosh","email":"data:image/png;base64,iVBORw0KGgoAAAANSUhEUgAAAZAAAAAyAQMAAABI0h/eAAAABlBMVEX///8AAABVwtN+AAAACXBIWXMAAA7EAAAOxAGVKw4bAAABDUlEQVRIie3RsWrDMBCA4QsGdTmTVSEQv4JMwFNBr2JNWZq1UwbRgjt2DvQhUgqGQgeBIF4KWQ0dbAi0a6BjXVpJzmiZjh30j0Yflu4AQqF/GIHonZ46vWBN9fGFnea8ObSQb0Ds1DCZAslm20IvWY1MP7CjKCgyOO1BPMphMpNI5rHUYlcj6I595sSQyfYGxJPnYkz1ZOXI1ZlEsQRR+smFecLPumzuWkPeOEnuW0de/OR6jkSvyxqYJZOCTuUo4QozS1aZJcgOhqCKvscv5kjOe1JZkkfx2PN1P+T02Q7ZEDdkjXu6TKWHVLdulYlZZavtKknymh5xc7lgyvObaOijOUw950OhUCj0l34BTgVvmtUGXgUAAAAASUVORK5CYII=","orcid":"","institution":"University of Westminster","correspondingAuthor":true,"prefix":"","firstName":"Anjali","middleName":"","lastName":"Ghosh","suffix":""},{"id":621719825,"identity":"56920503-4b69-4ff2-941e-7239ce2458a3","order_by":1,"name":"Schneider Fernandes","email":"","orcid":"","institution":"University of Westminster","correspondingAuthor":false,"prefix":"","firstName":"Schneider","middleName":"","lastName":"Fernandes","suffix":""}],"badges":[],"createdAt":"2026-03-15 17:39:24","currentVersionCode":1,"declarations":"","doi":"10.21203/rs.3.rs-9130366/v1","doiUrl":"https://doi.org/10.21203/rs.3.rs-9130366/v1","draftVersion":[],"editorialEvents":[],"editorialNote":"","failedWorkflow":false,"files":[{"id":106994410,"identity":"590489a2-d4f0-4bd2-9b26-9e897218f26b","added_by":"auto","created_at":"2026-04-15 15:08:22","extension":"pdf","order_by":1,"title":"","display":"","copyAsset":false,"role":"manuscript-pdf","size":1454065,"visible":true,"origin":"","legend":"","description":"","filename":"AGPhenotypicinvestigationofthecomplexmechanismsofbiofilmformationandroleofquorumsensingmoleculesinSalmonellaentericaserovars11121.pdf","url":"https://assets-eu.researchsquare.com/files/rs-9130366/v1_covered_9bd151a4-25f2-4fe0-9982-e8e8df564cba.pdf"}],"financialInterests":"No competing interests reported.","formattedTitle":"Dissecting the Phenotypic Complexity of Biofilm Development and Quorum Sensing in Salmonella enterica Serovars","fulltext":[],"fulltextSource":"","fullText":"","funders":[],"hasAdminPriorityOnWorkflow":false,"hasManuscriptDocX":false,"hasOptedInToPreprint":true,"hasPassedJournalQc":"","hasAnyPriority":false,"hideJournal":false,"highlight":"","institution":"","isAcceptedByJournal":false,"isAuthorSuppliedPdf":true,"isDeskRejected":"","isHiddenFromSearch":false,"isInQc":false,"isInWorkflow":false,"isPdf":true,"isPdfUpToDate":true,"isWithdrawnOrRetracted":false,"journal":{"display":true,"email":"[email protected]","identity":"bulletin-of-the-national-research-centre","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":false,"externalIdentity":"bnrc","sideBox":"Learn more about [Bulletin of the National Research Centre](https://BNRC.springeropen.com)","snPcode":"42269","submissionUrl":"https://submission.springernature.com/new-submission/42269/3","title":"Bulletin of the National Research Centre","twitterHandle":"","acdcEnabled":true,"dfaEnabled":true,"editorialSystem":"stoa","reportingPortfolio":"Springer Open","inReviewEnabled":true,"inReviewRevisionsEnabled":true},"keywords":"N-acyl homoserine lactone (AHL), Salmonella Typhimurium 14028, Salmonella Typhimurium Wildtype and Salmonella Enteritidis UCH, Salmonella Virchow, C8HSL, Farnesol, CV assay, fluorescence microscopy, antibiotic susceptibility test, Biofilm formation, quorum sensing, quorum quenching and natural compounds","lastPublishedDoi":"10.21203/rs.3.rs-9130366/v1","lastPublishedDoiUrl":"https://doi.org/10.21203/rs.3.rs-9130366/v1","license":{"name":"CC BY 4.0","url":"https://creativecommons.org/licenses/by/4.0/"},"manuscriptAbstract":"\u003cp\u003eThis study investigates the mechanisms of biofilm formation and quorum sensing in \u003cem\u003eSalmonella enterica\u003c/em\u003e serovars, with particular emphasis on their implications for clinical safety and public health. The ability of this bacterium to form biofilms on various surfaces poses significant challenges for control and eradication efforts, contributing to persistent infections and enhanced antimicrobial resistance. The study explores quorum quenching strategies to inhibit biofilm formation through certain treatments. The research employes crystal violet staining, bacterial growth curve analysis, and fluorescence microscopy to evaluate the effects of quorum sensing (QS) and natural product-based quorum quenching strategies on biofilm formation on abiotic (polystyrene) surfaces. Short- to medium-chain acyl-homoserine lactones (AHLs), particularly C8-HSL were assessed for their role in promoting biofilm development, alongside plant-derived and dietary natural products with reported antimicrobial activity, including aloe emodin (from \u003cem\u003eAloe vera\u003c/em\u003e), epigallocatechin gallate (EGCG) (from green tea, \u003cem\u003eCamellia sinensis\u003c/em\u003e), thymoquinone from (\u003cem\u003eNigella sativa)\u003c/em\u003e garlic extract (active compound \u003cem\u003eAllium sativum\u003c/em\u003e), and turmeric extract (active extract \u003cem\u003eCurcuma longa\u003c/em\u003e). Results demonstrated that C8-HSL significantly enhanced biofilm formation (Figs.\u0026nbsp;2\u0026ndash;4; Tables\u0026nbsp;1\u0026ndash;2), with 0.2 mM identified as the most effective concentration (Fig.\u0026nbsp;4). In contrast, the tested natural products exhibited quorum quenching and anti-biofilm activity, significantly reducing biofilm biomass and structural integrity (Figs.\u0026nbsp;5 and 6). Fluorescence microscopy further supported these findings by demonstrating enhanced biofilm formation in the presence of quorum sensing molecules and disrupted biofilm architecture following quorum quenching treatment (Figs.\u0026nbsp;7 and 8). These findings highlight the central role of quorum sensing in \u003cem\u003eSalmonella enterica\u003c/em\u003e pathogenicity and demonstrate the potential of plant-derived and dietary natural products as alternative quorum quenching agents for biofilm control. The study suggests that further research, including the use of genetically engineered mutants, is required to elucidate the specific genetic pathways involved in QS-mediated biofilm regulation. Overall, this research advances understanding of biofilm formation and quorum sensing in \u003cem\u003eSalmonella enterica\u003c/em\u003e and supports the development of novel strategies to manage persistent infections and improve food safety.\u003c/p\u003e","manuscriptTitle":"Dissecting the Phenotypic Complexity of Biofilm Development and Quorum Sensing in Salmonella enterica Serovars","msid":"","msnumber":"","nonDraftVersions":[{"code":1,"date":"2026-04-15 11:07:31","doi":"10.21203/rs.3.rs-9130366/v1","editorialEvents":[{"type":"communityComments","content":0},{"type":"decision","content":"Revision requested","date":"2026-04-30T11:10:34+00:00","index":"","fulltext":""},{"type":"editorInvitedReview","content":"","date":"2026-04-27T05:40:10+00:00","index":"hide","fulltext":""},{"type":"editorInvitedReview","content":"","date":"2026-04-26T19:24:07+00:00","index":"hide","fulltext":""},{"type":"editorInvitedReview","content":"","date":"2026-04-24T06:23:28+00:00","index":"hide","fulltext":""},{"type":"editorInvitedReview","content":"","date":"2026-04-22T13:23:15+00:00","index":"hide","fulltext":""},{"type":"editorInvitedReview","content":"","date":"2026-04-19T03:46:17+00:00","index":"hide","fulltext":""},{"type":"reviewerAgreed","content":"253868148100305422307104449845983021628","date":"2026-04-12T14:42:13+00:00","index":"hide","fulltext":""},{"type":"reviewerAgreed","content":"252506855791281300202749766199569898349","date":"2026-04-09T18:55:10+00:00","index":"hide","fulltext":""},{"type":"reviewerAgreed","content":"332379369758332095703286392108704004936","date":"2026-04-09T14:47:31+00:00","index":"hide","fulltext":""},{"type":"reviewerAgreed","content":"293508203879140274620195207477841795581","date":"2026-04-08T07:46:34+00:00","index":"hide","fulltext":""},{"type":"reviewerAgreed","content":"46425164602671120860328524904268020437","date":"2026-04-08T05:32:59+00:00","index":"hide","fulltext":""},{"type":"reviewerAgreed","content":"18374825670563173404848361415112071259","date":"2026-04-08T05:22:23+00:00","index":"hide","fulltext":""},{"type":"reviewerAgreed","content":"89504530491852657584841980725668084191","date":"2026-04-08T05:15:09+00:00","index":"hide","fulltext":""},{"type":"reviewersInvited","content":"","date":"2026-04-08T05:12:39+00:00","index":"","fulltext":""},{"type":"editorAssigned","content":"","date":"2026-04-07T11:57:34+00:00","index":"","fulltext":""},{"type":"checksComplete","content":"","date":"2026-04-07T07:42:25+00:00","index":"","fulltext":""},{"type":"submitted","content":"Bulletin of the National Research Centre","date":"2026-04-06T20:39:44+00:00","index":"","fulltext":""}],"status":"published","journal":{"display":true,"email":"[email protected]","identity":"bulletin-of-the-national-research-centre","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":false,"externalIdentity":"bnrc","sideBox":"Learn more about [Bulletin of the National Research Centre](https://BNRC.springeropen.com)","snPcode":"42269","submissionUrl":"https://submission.springernature.com/new-submission/42269/3","title":"Bulletin of the National Research Centre","twitterHandle":"","acdcEnabled":true,"dfaEnabled":true,"editorialSystem":"stoa","reportingPortfolio":"Springer Open","inReviewEnabled":true,"inReviewRevisionsEnabled":true}}],"origin":"","ownerIdentity":"cfda7649-40f5-4341-9482-99f14a99d5d7","owner":[],"postedDate":"April 15th, 2026","published":true,"recentEditorialEvents":[{"type":"decision","content":"Revision requested","date":"2026-04-30T11:10:34+00:00","index":"","fulltext":""}],"rejectedJournal":[],"revision":"","amendment":"","status":"in-revision","subjectAreas":[],"tags":[],"updatedAt":"2026-04-30T11:24:13+00:00","versionOfRecord":[],"versionCreatedAt":"2026-04-15 11:07:31","video":"","vorDoi":"","vorDoiUrl":"","workflowStages":[]},"version":"v1","identity":"rs-9130366","journalConfig":"researchsquare"},"__N_SSP":true},"page":"/article/[identity]/[[...version]]","query":{"redirect":"/article/rs-9130366","identity":"rs-9130366","version":["v1"]},"buildId":"XKTyCvWXoU3ODBz1xrDgd","isFallback":false,"isExperimentalCompile":false,"dynamicIds":[84888],"gssp":true,"scriptLoader":[]}