Targeting STEC-Induced Edema Disease in Weaned Piglets: Prophylactic Oral Phage P-GXEC-L2P5 Mitigates Vascular Endothelial Injury

Targeting STEC-Induced Edema Disease in Weaned Piglets: Prophylactic Oral Phage P-GXEC-L2P5 Mitigates Vascular Endothelial Injury | 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 Targeting STEC-Induced Edema Disease in Weaned Piglets: Prophylactic Oral Phage P-GXEC-L2P5 Mitigates Vascular Endothelial Injury Runwen Ma, Shuming Tang, Jialing Zeng, Yixian Wei, Xun Li, Xiaoye Wang This is a preprint; it has not been peer reviewed by a journal. https://doi.org/ 10.21203/rs.3.rs-6754528/v1 This work is licensed under a CC BY 4.0 License Status: Published Journal Publication published 17 Dec, 2025 Read the published version in Veterinary Research → Version 1 posted You are reading this latest preprint version Abstract Edema disease (ED), a frequently fatal disease in weaned piglets, is caused by Shiga toxin-producing Escherichia coli (STEC). With the increasing emergence of antibiotic-resistant E. coli strains, bacteriophage therapy has been considered a promising alternative to prevent and control bacterial diseases in swine farms. Using a porcine infection model, the present study evaluated the prophylactic effect of bacteriophage P-GXEC-L2P5 administered by oral gavage against ED. A novel phage named P-GXEC-L2P5 was isolated using a multidrug-resistant (MDR) STEC named GXEC-STL2 as host strain. Phage P-GXEC-L2P5 was identified as a member of Dhillonvirus, Caudoviricetes with an 88,607 bp genome and possessed a short latent period (10 min), moderate pH stability (5–10), and appropriate thermal tolerance (4–60°C). Piglets pretreated with phage P-GXEC-L2P5 showed no apparent clinical signs such as eyelid edema or neurological symptoms after GXEC-STL2 challenge. B-scan ultrasonographic examination revealed no significant hydronephrosis. Necropsy findings indicated only mild intestinal congestion with no other gross pathological lesions observed. Histopathological examination demonstrated no significant differences compared to non-infected controls. Phage P-GXEC-L2P5 significantly reduced fecal shedding of STEC (P<0.05) in piglets, with a concurrent marked decrease (P<0.01) in Stx2e concentrations detected in serum, cerebral cortex, kidney, and small intestine. The cerebral cortex, kidney, and small intestine displayed significantly lower mRNA expression of Gb4 (Stx2e receptor) (P<0.05), while vascular endothelia exhibited a significant increase (P<0.05) in both FITC-WGA fluorescence intensity and mRNA expression of endothelial integrity-associated factors (connexin43, vinculin, zonula occludens-1). Furthermore, gut microbiota analysis found that phage treatment preserved the diversity and abundance of jejunal microbiota. In conclusion, phage P-GXEC-L2P5 effectively prevented STEC-induced ED by reducing STEC load and Stx2e levels, while mitigating vascular permeability. Bacteriophage STEC Edema disease Prophylactic therapy Full Text Additional Declarations No competing interests reported. Supplementary Files Additionalfile1Virulencegenesprimerinformation.docx Additionalfile2Clinicalsymptomscoringcriteria.docx Additionalfile3TheqPCRprimersandreactionconditionsforvascularendothelialintegrityassociatedfactors.docx Additionalfile4ResultsoftheKBdiskdiffusionmethodfordeterminingthesusceptibilityofGXECSTL2toantibiotics..docx Additionalfile5HostspectrumofphagePGXECL2P5to58Escherichiacoliisolates2preservedSalmonellaisolates..docx FigureS1.tif Cite Share Download PDF Status: Published Journal Publication published 17 Dec, 2025 Read the published version in Veterinary Research → 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. 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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-6754528","acceptedTermsAndConditions":true,"allowDirectSubmit":true,"archivedVersions":[],"articleType":"Research Article","associatedPublications":[],"authors":[{"id":482541341,"identity":"0a338b36-ed3c-44b8-81e5-c0773cd0d5a6","order_by":0,"name":"Runwen Ma","email":"","orcid":"","institution":"College of Animal Science and Technology, Guangxi University","correspondingAuthor":false,"prefix":"","firstName":"Runwen","middleName":"","lastName":"Ma","suffix":""},{"id":482541342,"identity":"61b8ea01-c889-4608-846a-83d10403d629","order_by":1,"name":"Shuming Tang","email":"","orcid":"","institution":"College of Animal Science and 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Injury","fulltext":[],"fulltextSource":"","fullText":"","funders":[],"hasAdminPriorityOnWorkflow":false,"hasManuscriptDocX":false,"hasOptedInToPreprint":true,"hasPassedJournalQc":"","hasAnyPriority":false,"hideJournal":true,"highlight":"","institution":"","isAcceptedByJournal":true,"isAuthorSuppliedPdf":true,"isDeskRejected":"","isHiddenFromSearch":false,"isInQc":false,"isInWorkflow":false,"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":"Bacteriophage, STEC, Edema disease, Prophylactic therapy","lastPublishedDoi":"10.21203/rs.3.rs-6754528/v1","lastPublishedDoiUrl":"https://doi.org/10.21203/rs.3.rs-6754528/v1","license":{"name":"CC BY 4.0","url":"https://creativecommons.org/licenses/by/4.0/"},"manuscriptAbstract":"Edema disease (ED), a frequently fatal disease in weaned piglets, is caused by Shiga toxin-producing Escherichia coli (STEC). With the increasing emergence of antibiotic-resistant E. coli strains, bacteriophage therapy has been considered a promising alternative to prevent and control bacterial diseases in swine farms. Using a porcine infection model, the present study evaluated the prophylactic effect of bacteriophage P-GXEC-L2P5 administered by oral gavage against ED.\nA novel phage named P-GXEC-L2P5 was isolated using a multidrug-resistant (MDR) STEC named GXEC-STL2 as host strain. Phage P-GXEC-L2P5 was identified as a member of Dhillonvirus, Caudoviricetes with an 88,607 bp genome and possessed a short latent period (10 min), moderate pH stability (5–10), and appropriate thermal tolerance (4–60°C). Piglets pretreated with phage P-GXEC-L2P5 showed no apparent clinical signs such as eyelid edema or neurological symptoms after GXEC-STL2 challenge. B-scan ultrasonographic examination revealed no significant hydronephrosis. Necropsy findings indicated only mild intestinal congestion with no other gross pathological lesions observed. Histopathological examination demonstrated no significant differences compared to non-infected controls. Phage P-GXEC-L2P5 significantly reduced fecal shedding of STEC (P\u003c0.05) in piglets, with a concurrent marked decrease (P\u003c0.01) in Stx2e concentrations detected in serum, cerebral cortex, kidney, and small intestine. The cerebral cortex, kidney, and small intestine displayed significantly lower mRNA expression of Gb4 (Stx2e receptor) (P\u003c0.05), while vascular endothelia exhibited a significant increase (P\u003c0.05) in both FITC-WGA fluorescence intensity and mRNA expression of endothelial integrity-associated factors (connexin43, vinculin, zonula occludens-1). Furthermore, gut microbiota analysis found that phage treatment preserved the diversity and abundance of jejunal microbiota.\nIn conclusion, phage P-GXEC-L2P5 effectively prevented STEC-induced ED by reducing STEC load and Stx2e levels, while mitigating vascular permeability.","manuscriptTitle":"Targeting STEC-Induced Edema Disease in Weaned Piglets: Prophylactic Oral Phage P-GXEC-L2P5 Mitigates Vascular Endothelial Injury","msid":"","msnumber":"","nonDraftVersions":[{"code":1,"date":"2025-07-10 04:21:47","doi":"10.21203/rs.3.rs-6754528/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":"20a1392b-753e-4d1b-9708-146fbe1fe2c4","owner":[],"postedDate":"July 10th, 2025","published":true,"recentEditorialEvents":[],"rejectedJournal":[],"revision":"","amendment":"","status":"posted","subjectAreas":[],"tags":[],"updatedAt":"2025-12-22T16:02:21+00:00","versionOfRecord":{"articleIdentity":"rs-6754528","link":"https://doi.org/10.1186/s13567-025-01683-w","journal":{"identity":"veterinary-research","isVorOnly":false,"title":"Veterinary Research"},"publishedOn":"2025-12-17 15:57:18","publishedOnDateReadable":"December 17th, 2025"},"versionCreatedAt":"2025-07-10 04:21:47","video":"","vorDoi":"10.1186/s13567-025-01683-w","vorDoiUrl":"https://doi.org/10.1186/s13567-025-01683-w","workflowStages":[]},"version":"v1","identity":"rs-6754528","journalConfig":"researchsquare"},"__N_SSP":true},"page":"/article/[identity]/[[...version]]","query":{"redirect":"/article/rs-6754528","identity":"rs-6754528","version":["v1"]},"buildId":"8U1c8b4HqxoKbykW_rLl7","isFallback":false,"isExperimentalCompile":false,"dynamicIds":[84888],"gssp":true,"scriptLoader":[]}