Galangin Targets Type IV Pilus to Attenuate Clostridium perfringens Pathogenicity in Broilers Chickens

Galangin Targets Type IV Pilus to Attenuate Clostridium perfringens Pathogenicity in Broilers Chickens | 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 Galangin Targets Type IV Pilus to Attenuate Clostridium perfringens Pathogenicity in Broilers Chickens Tingting Chen, Zeyu Song, Bingyu Hou, Xiaonan Zhao, Yonglin Zhou, and 2 more This is a preprint; it has not been peer reviewed by a journal. https://doi.org/ 10.21203/rs.3.rs-4457926/v2 This work is licensed under a CC BY 4.0 License Status: Posted Version 2 posted You are reading this latest preprint version Show more versions Abstract Clostridium perfringens ( C. perfringens ) induces severe clinical manifestations including necrotic enteritis (NE), enterotoxemia, foodborne illness, and gas gangrene, posing substantial threats to public health and livestock industries. Our study focused on the type IV pilus (TFP) of C. perfringens to elucidate the molecular mechanisms underlying natural compound-mediated bacterial inhibition. Firstly, we identified galangin as a candidate through phenotypic screening of TFP-mediated gliding motility and biofilm formation. Subsequently, the in vitro evaluations encompassed bacterial growth kinetics, minimum inhibitory concentration (MIC) determination, cytotoxicity profiling, and cell adhesion assays. Then, the NE-broilers model was established to assess galangin’s therapeutic potential, incorporating qPCR analysis of virS / virR two-component regulatory genes and TFP-encoding genes ( pilA , pilC , pilD , pilT and pilM ), complemented by transmission electron microscopy (TEM) for TFP ultrastructural characterization. Galangin(8 μg/mL) significantly attenuated bacterial motility (29.79%) and biofilm formation (24.85%). It reduced Caco-2 cell adhesion by 45.72% ( p<0.05 ) while maintaining cellular viability. In vivo administration (20 mg/kg galangin) enhanced broiler average weight (galangin group 99.86 g vs 52.68 g Infection group) and improved feed conversion ratio (3.09 vs 5.76). Histopathological analysis demonstrated reduction in ileal lesion, accompanied by 0.78-log CFU/g bacterial load decrease and normalized inflammatory cytokine levels (IL-1β 38.92%, TNF-α 31.53%, IL-6 31.13%). TEM revealed complete TFP structural disintegration, while qPCR confirmed significant downregulation of virR / virS ( virR :1.89-fold, virS :1.34-fold) and TFP genes ( pilA :1.27-fold, pilD :1.43-fold, pilT :2.11-fold, pilM :1.95-fold). This study establishes galangin as a novel TFP-targeted anti-virulence agent against C. perfringens , providing mechanistic insights and therapeutic validation for NE prevention. Small Animal Medicine Clinical Pharmacology Zoonoses Clostridium perfringens galangin Type IV pilus anti-virulence Figures Figure 1 Figure 2 Figure 3 Figure 4 Figure 5 Figure 6 Figure 7 Figure 8 Full Text Additional Declarations The authors declare no competing interests. Cite Share Download PDF Status: Posted Version 2 posted You are reading this latest preprint version Show more versions 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-4457926","acceptedTermsAndConditions":true,"allowDirectSubmit":true,"archivedVersions":[],"articleType":"Research Article","associatedPublications":[],"authors":[{"id":558309807,"identity":"41a5f96f-5a64-432d-a142-6f2082295820","order_by":0,"name":"Tingting Chen","email":"","orcid":"","institution":"Shandong Academy of Agricultural Sciences","correspondingAuthor":false,"prefix":"","firstName":"Tingting","middleName":"","lastName":"Chen","suffix":""},{"id":558309808,"identity":"c77855c9-6350-4bd6-8045-c9552a9fa0fe","order_by":1,"name":"Zeyu Song","email":"","orcid":"","institution":"Jilin University","correspondingAuthor":false,"prefix":"","firstName":"Zeyu","middleName":"","lastName":"Song","suffix":""},{"id":558309809,"identity":"2f0a6379-3a04-4ad8-b43c-7145180a150f","order_by":2,"name":"Bingyu Hou","email":"","orcid":"","institution":"Shandong Academy of Agricultural Sciences","correspondingAuthor":false,"prefix":"","firstName":"Bingyu","middleName":"","lastName":"Hou","suffix":""},{"id":558309810,"identity":"5023aba9-7b0e-44ea-8bf7-41c5010a8485","order_by":3,"name":"Xiaonan Zhao","email":"","orcid":"","institution":"Shandong Academy of Agricultural Sciences","correspondingAuthor":false,"prefix":"","firstName":"Xiaonan","middleName":"","lastName":"Zhao","suffix":""},{"id":558309812,"identity":"d3851511-ac78-465e-a84d-7800b12e931f","order_by":4,"name":"Yonglin Zhou","email":"","orcid":"","institution":"Ningxia University","correspondingAuthor":false,"prefix":"","firstName":"Yonglin","middleName":"","lastName":"Zhou","suffix":""},{"id":558309811,"identity":"161644de-171b-4b8b-a544-d0fc5c2274cd","order_by":5,"name":"Xiaolei Shang","email":"","orcid":"","institution":"Shandong Jinzhuji Pharmaceutical Co., Ltd","correspondingAuthor":false,"prefix":"","firstName":"Xiaolei","middleName":"","lastName":"Shang","suffix":""},{"id":558309813,"identity":"71c07f62-6cf3-46bb-98ea-e9e416a03e1a","order_by":6,"name":"Qianghua Lv","email":"data:image/png;base64,iVBORw0KGgoAAAANSUhEUgAAAZAAAAAyAQMAAABI0h/eAAAABlBMVEX///8AAABVwtN+AAAACXBIWXMAAA7EAAAOxAGVKw4bAAABCklEQVRIiWNgGAWjYBACPiA+AMQJDAyMjQ8+VLDJgEQl8GlhQ2hhbjaccYaNhygtDBAt7G3SvG0MRGiRSN544OeO2jyD4wcbpHnn8fEYHGA+eJuHwS4Pt5a0goO9Z44XG5xJbDCcu40NqIUt2ZqHIbkYpxaeMwYHeNuOJW44kNiQ8BashcdMmocByMWj5eBfkJbzDxsO8M4BaeH/hl8Le4/BYd62msQNNxIbG3kbwLawEdDSVnBYtu1A4swbD5sZZxxj45E8zGZsOccgGacWfmbmzR/fttUl9p1Pf/7jQ80xOb7jzQ9vvKmww6kFCAyA+DCMcwwYpTBB/FrqYJwavEpHwSgYBaNgZAIAPfBdICe4HlIAAAAASUVORK5CYII=","orcid":"","institution":"Shandong Academy of Agricultural Sciences","correspondingAuthor":true,"prefix":"","firstName":"Qianghua","middleName":"","lastName":"Lv","suffix":""}],"badges":[],"createdAt":"2024-05-22 03:29:46","currentVersionCode":2,"declarations":{"humanSubjects":false,"vertebrateSubjects":false,"conflictsOfInterestStatement":false,"humanSubjectEthicalGuidelines":false,"humanSubjectConsent":false,"humanSubjectClinicalTrial":false,"humanSubjectCaseReport":false,"vertebrateSubjectEthicalGuidelines":false},"doi":"10.21203/rs.3.rs-4457926/v2","doiUrl":"https://doi.org/10.21203/rs.3.rs-4457926/v2","draftVersion":[],"editorialEvents":[],"editorialNote":"","failedWorkflow":false,"files":[{"id":98418435,"identity":"de2c42a6-a6d5-4135-871a-6716d41f3ea5","added_by":"auto","created_at":"2025-12-17 15:12:54","extension":"jpg","order_by":1,"title":"Figure 1","display":"","copyAsset":false,"role":"figure","size":513616,"visible":true,"origin":"","legend":"\u003cp\u003e\u003cstrong\u003eInhibitory effects of galangin against gliding motility and biofilm formation.\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003e(A) Chemical structure of galangin. (B) Galangin inhibits the gliding motility of \u003cem\u003eC. perfringens \u003c/em\u003eATCC13124. (C) The gliding motility ratio of \u003cem\u003eC. perfringens\u003c/em\u003eATCC13124 with different concentrations of galangin. The WT group without galangin was set as 100%. (D) Galangin inhibits \u003cem\u003eC. perfringens\u003c/em\u003e ATCC13124 biofilm formation. The group without galangin was set as 100%. The data are shown as the mean ± SEM of three independent preparations. * \u003cem\u003ep \u003c/em\u003e\u0026lt; 0.05; ** \u003cem\u003ep \u003c/em\u003e\u0026lt; 0.01.\u003c/p\u003e","description":"","filename":"Figure1.jpg","url":"https://assets-eu.researchsquare.com/files/rs-4457926/v2/8d1a121e8c3bad24c9269fa2.jpg"},{"id":98441115,"identity":"5de1e93b-575c-4b8c-b548-dd00b9a2164d","added_by":"auto","created_at":"2025-12-17 17:04:54","extension":"jpg","order_by":2,"title":"Figure 2","display":"","copyAsset":false,"role":"figure","size":717265,"visible":true,"origin":"","legend":"\u003cp\u003e\u003cstrong\u003eNonbacteriostatic mechanism of galangin.\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003e(A) Growth curves of \u003cem\u003eC. perfringens\u003c/em\u003e treated with different concentrations of galangin (0~64 μg/ml). To clarify the effect of galangin on the growth of \u003cem\u003eC. perfringens\u003c/em\u003e, the absorbance of the culture was measured at 600 nm. (B) MIC of galangin against \u003cem\u003eC. perfringens \u003c/em\u003eATCC13124\u003cem\u003e. \u003c/em\u003eBacterial colony formation on agar plates was used to measure the MIC.\u003c/p\u003e","description":"","filename":"Figure2.jpg","url":"https://assets-eu.researchsquare.com/files/rs-4457926/v2/fd09cd24e072e90485b7bf50.jpg"},{"id":98441825,"identity":"ed210050-d8c7-4faa-893e-d59dd2501950","added_by":"auto","created_at":"2025-12-17 17:05:53","extension":"jpg","order_by":3,"title":"Figure 3","display":"","copyAsset":false,"role":"figure","size":601059,"visible":true,"origin":"","legend":"\u003cp\u003e\u003cstrong\u003eGalangin inhibits the adhesion of \u003c/strong\u003e\u003cem\u003e\u003cstrong\u003eC. perfringens\u003c/strong\u003e\u003c/em\u003e\u003cstrong\u003e to Caco-2 cells without cytotoxicity. \u003c/strong\u003e(A) The cytotoxicity of galangin on Caco-2 cells at different concentrations. (B) Inhibitory effect of galangin on the adhesion of \u003cem\u003eC. perfringens\u003c/em\u003e. Caco-2 cells were infected with\u003cem\u003e C. perfringens\u003c/em\u003e ATCC13124 pretreated with galangin. All data are shown as the mean ± SD from three independent experiments. *\u003cem\u003ep \u003c/em\u003e\u0026lt; 0.05; **\u003cem\u003ep \u003c/em\u003e\u0026lt; 0.01 compared to the control group.\u003c/p\u003e","description":"","filename":"Figure3.jpg","url":"https://assets-eu.researchsquare.com/files/rs-4457926/v2/76d72e42f01cd9c4028ed924.jpg"},{"id":98441824,"identity":"7ed1a3db-ec6b-472f-b71d-3b505eb27385","added_by":"auto","created_at":"2025-12-17 17:05:53","extension":"jpg","order_by":4,"title":"Figure 4","display":"","copyAsset":false,"role":"figure","size":538365,"visible":true,"origin":"","legend":"\u003cp\u003e\u003cstrong\u003eGalangin inhibits the synthesis of TFP.\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003e(A) The pili morphology of \u003cem\u003eC. perfringens\u003c/em\u003e ATCC13124 treated with galangin under TEM. (B) The mRNA expression level of TFP coding genes and two-component regulatory genes with different concentrations of galangin. *\u003cem\u003ep \u003c/em\u003e\u0026lt; 0.05.\u003c/p\u003e","description":"","filename":"Figure4.jpg","url":"https://assets-eu.researchsquare.com/files/rs-4457926/v2/a1a90a7ad8109c524496590e.jpg"},{"id":98418437,"identity":"1095ebc3-c227-4543-b6ae-eb8568a0d612","added_by":"auto","created_at":"2025-12-17 15:12:54","extension":"jpg","order_by":5,"title":"Figure 5","display":"","copyAsset":false,"role":"figure","size":364453,"visible":true,"origin":"","legend":"\u003cp\u003e\u003cstrong\u003eEffect of galangin on the performance of broilers infected with\u003c/strong\u003e\u003cem\u003e\u003cstrong\u003e C. perfringens \u003c/strong\u003e\u003c/em\u003e\u003cstrong\u003eATCC13124.\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003e(A) Average weights of broilers at 21 and 30 days. (B) The feed conversion rate of broilers during the experiment. *\u003cem\u003e p\u003c/em\u003e\u0026lt;0.05; **\u003cem\u003ep\u003c/em\u003e\u0026lt;0.01; NS, not significant.\u003c/p\u003e","description":"","filename":"Figure5.jpg","url":"https://assets-eu.researchsquare.com/files/rs-4457926/v2/c340162266936b7f658c3791.jpg"},{"id":98418440,"identity":"ff9e830d-acc5-4dc9-84f6-f1743fcd14cc","added_by":"auto","created_at":"2025-12-17 15:12:54","extension":"jpg","order_by":6,"title":"Figure 6","display":"","copyAsset":false,"role":"figure","size":1212405,"visible":true,"origin":"","legend":"\u003cp\u003e\u003cstrong\u003eEffect of galangin on pathological ileal damage in broilers infected with \u003c/strong\u003e\u003cem\u003e\u003cstrong\u003eC. perfringens.\u003c/strong\u003e\u003c/em\u003e\u003c/p\u003e\n\u003cp\u003e(A) Intestinal bleeding in broilers with NE. (B) Pathological ileal sections in broilers with NE. (C~E) The villus height, crypt depth and VCR of broilers with NE. All data are shown as the mean ± SEM from three independent experiments. * \u003cem\u003ep\u003c/em\u003e \u0026lt; 0.05, ** \u003cem\u003ep \u003c/em\u003e\u0026lt; 0.01; NS, not significant.\u003c/p\u003e","description":"","filename":"Figure6.jpg","url":"https://assets-eu.researchsquare.com/files/rs-4457926/v2/17e21f634947f79bf9aea53e.jpg"},{"id":98441662,"identity":"45cd7268-b753-4c44-9138-a398a7fd295d","added_by":"auto","created_at":"2025-12-17 17:05:40","extension":"jpg","order_by":7,"title":"Figure 7","display":"","copyAsset":false,"role":"figure","size":633125,"visible":true,"origin":"","legend":"\u003cp\u003e\u003cstrong\u003eEffects of galangin on ileal colony colonization and release of inflammatory cytokines in broilers infected with \u003c/strong\u003e\u003cem\u003e\u003cstrong\u003eC. perfringens\u003c/strong\u003e\u003c/em\u003e\u003cstrong\u003e.\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003e(A) Ileal colony colonization of broilers in each group. (B~D) Inflammatory cytokines IL-1β, IL-6 and TNF-α in each group. All data are shown as the mean ± SEM from three independent experiments. * \u003cem\u003ep\u003c/em\u003e \u0026lt; 0.05; **\u003cem\u003e p\u003c/em\u003e \u0026lt; 0.01; NS, not significant.\u003c/p\u003e","description":"","filename":"Figure7.jpg","url":"https://assets-eu.researchsquare.com/files/rs-4457926/v2/bea75d961570b2258df99037.jpg"},{"id":98622413,"identity":"c3633d76-9855-4bd5-ace7-9ee3e4769921","added_by":"auto","created_at":"2025-12-19 16:54:33","extension":"jpg","order_by":8,"title":"Figure 8","display":"","copyAsset":false,"role":"figure","size":881490,"visible":true,"origin":"","legend":"\u003cp\u003e\u003cstrong\u003eGalangin improves the expression of genes encoding tight junction proteins and inhibits ileal mucosal factors in \u003c/strong\u003e\u003cem\u003e\u003cstrong\u003eC. perfringens\u003c/strong\u003e\u003c/em\u003e\u003cstrong\u003e-infected broilers.\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003e(A~B) Expression of the tight junction-coding genes \u003cem\u003eOccludin\u003c/em\u003e and \u003cem\u003eClaudin\u003c/em\u003e in broilers infected with \u003cem\u003eC. perfringens \u003c/em\u003ein each group. (C~D) Expression of \u003cem\u003eTLR2\u003c/em\u003e and \u003cem\u003eTLR4 \u003c/em\u003ein broilers infected with \u003cem\u003eC. perfringens \u003c/em\u003ein each group. (E) Expression of \u003cem\u003eMUC2 \u003c/em\u003ein broilers infected with \u003cem\u003eC. perfringens \u003c/em\u003ein each group. All data are shown as the mean ± SEM from three independent experiments. * \u003cem\u003ep \u003c/em\u003e\u0026lt; 0.05; **\u003cem\u003e p\u003c/em\u003e \u0026lt; 0.01; NS, not significant.\u003c/p\u003e","description":"","filename":"Figure8.jpg","url":"https://assets-eu.researchsquare.com/files/rs-4457926/v2/02b0349f7d99ee55e8a9148e.jpg"},{"id":98631201,"identity":"8b6dca04-667f-4570-9edc-38108d7205d8","added_by":"auto","created_at":"2025-12-19 17:19:23","extension":"pdf","order_by":1,"title":"","display":"","copyAsset":false,"role":"manuscript-pdf","size":6218468,"visible":true,"origin":"","legend":"","description":"","filename":"Manuscriptpdf.pdf","url":"https://assets-eu.researchsquare.com/files/rs-4457926/v2_covered_7982c879-5233-469f-b473-cd552354c2db.pdf"}],"financialInterests":"The authors declare no competing interests.","formattedTitle":"\u003cp\u003e\u003cstrong\u003eGalangin Targets Type IV Pilus to Attenuate \u003c/strong\u003e\u003cem\u003e\u003cstrong\u003eClostridium perfringens\u003c/strong\u003e\u003c/em\u003e\u003cstrong\u003e Pathogenicity in Broilers Chickens\u003c/strong\u003e\u003c/p\u003e","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":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":"Clostridium perfringens; galangin; Type IV pilus; anti-virulence","lastPublishedDoi":"10.21203/rs.3.rs-4457926/v2","lastPublishedDoiUrl":"https://doi.org/10.21203/rs.3.rs-4457926/v2","license":{"name":"CC BY 4.0","url":"https://creativecommons.org/licenses/by/4.0/"},"manuscriptAbstract":"\u003cp\u003e\u003cem\u003eClostridium perfringens\u003c/em\u003e (\u003cem\u003eC. perfringens\u003c/em\u003e) induces severe clinical manifestations including necrotic enteritis (NE), enterotoxemia, foodborne illness, and gas gangrene, posing substantial threats to public health and livestock industries. Our study focused on the type IV pilus (TFP) of \u003cem\u003eC. perfringens\u003c/em\u003e to elucidate the molecular mechanisms underlying natural compound-mediated bacterial inhibition. Firstly, we identified galangin as a candidate through phenotypic screening of TFP-mediated gliding motility and biofilm formation. Subsequently, the \u003cem\u003ein vitro\u003c/em\u003e evaluations encompassed bacterial growth kinetics, minimum inhibitory concentration (MIC) determination, cytotoxicity profiling, and cell adhesion assays. Then, the NE-broilers model was established to assess galangin’s therapeutic potential, incorporating qPCR analysis of \u003cem\u003evirS\u003c/em\u003e/\u003cem\u003evirR\u003c/em\u003e two-component regulatory genes and TFP-encoding genes (\u003cem\u003epilA\u003c/em\u003e, \u003cem\u003epilC\u003c/em\u003e, \u003cem\u003epilD\u003c/em\u003e, \u003cem\u003epilT\u003c/em\u003e and \u003cem\u003epilM\u003c/em\u003e), complemented by transmission electron microscopy (TEM) for TFP ultrastructural characterization. Galangin(8 μg/mL) significantly attenuated bacterial motility (29.79%) and biofilm formation (24.85%). It reduced Caco-2 cell adhesion by 45.72% (\u003cem\u003ep\u0026lt;0.05\u003c/em\u003e) while maintaining cellular viability. \u003cem\u003eIn vivo\u003c/em\u003eadministration (20 mg/kg galangin) enhanced broiler average weight (galangin group 99.86 g vs 52.68 g Infection group) and improved feed conversion ratio (3.09 vs 5.76). Histopathological analysis demonstrated reduction in ileal lesion, accompanied by 0.78-log CFU/g bacterial load decrease and normalized inflammatory cytokine levels (IL-1β 38.92%, TNF-α 31.53%, IL-6 31.13%). TEM revealed complete TFP structural disintegration, while qPCR confirmed significant downregulation of \u003cem\u003evirR\u003c/em\u003e/\u003cem\u003evirS \u003c/em\u003e(\u003cem\u003evirR\u003c/em\u003e:1.89-fold, \u003cem\u003evirS\u003c/em\u003e:1.34-fold) and TFP genes (\u003cem\u003epilA\u003c/em\u003e:1.27-fold, \u003cem\u003epilD\u003c/em\u003e:1.43-fold, \u003cem\u003epilT\u003c/em\u003e:2.11-fold, \u003cem\u003epilM\u003c/em\u003e:1.95-fold). This study establishes galangin as a novel TFP-targeted anti-virulence agent against \u003cem\u003eC. perfringens\u003c/em\u003e, providing mechanistic insights and therapeutic validation for NE prevention.\u003c/p\u003e","manuscriptTitle":"Galangin Targets Type IV Pilus to Attenuate Clostridium perfringens Pathogenicity in Broilers Chickens","msid":"","msnumber":"","nonDraftVersions":[{"code":2,"date":"2025-12-17 15:12:49","doi":"10.21203/rs.3.rs-4457926/v2","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}},{"code":1,"date":"2024-06-03 22:58:19","doi":"10.21203/rs.3.rs-4457926/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":"f79569e0-3c6a-4c0a-9520-f9517a44ae1e","owner":[],"postedDate":"December 17th, 2025","published":true,"recentEditorialEvents":[],"rejectedJournal":[],"revision":"","amendment":"","status":"posted","subjectAreas":[{"id":59729345,"name":"Small Animal Medicine"},{"id":59729346,"name":"Clinical Pharmacology"},{"id":59729347,"name":"Zoonoses"}],"tags":[],"updatedAt":"2024-06-05T16:55:36+00:00","versionOfRecord":[],"versionCreatedAt":"2025-12-17 15:12:49","video":"","vorDoi":"","vorDoiUrl":"","workflowStages":[]},"version":"v2","identity":"rs-4457926","journalConfig":"researchsquare"},"__N_SSP":true},"page":"/article/[identity]/[[...version]]","query":{"redirect":"/article/rs-4457926","identity":"rs-4457926","version":["v2"]},"buildId":"8U1c8b4HqxoKbykW_rLl7","isFallback":false,"isExperimentalCompile":false,"dynamicIds":[84888],"gssp":true,"scriptLoader":[]}