Effect of feeding Bacillus subtilis on intestinal bloat and microbiological structure in sows infected with Clostridium perfringens type A

Effect of feeding Bacillus subtilis on intestinal bloat and microbiological structure in sows infected with Clostridium perfringens type A | 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 Effect of feeding Bacillus subtilis on intestinal bloat and microbiological structure in sows infected with Clostridium perfringens type A Mengran Zhang, Aohang-Yu Yu, Chaochi Wang, Jiayue Bai, Chaojie Chen, and 1 more This is a preprint; it has not been peer reviewed by a journal. https://doi.org/ 10.21203/rs.3.rs-7359451/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 Bacillus subtilis is aerobic or facultatively anaerobic.After entering the gastrointestinal tract, spores colonize the gut, inhibiting the growth of harmful aerobic bacteria ( Escherichia coli , Streptococcus , Staphylococcus aureus ).However, it is not clear whether Bacillus subtilis can inhibit Clostridium perfringens type A infection. In this study, Bacillus subtilis PB6 was added to the diets of Clostridium perfringens type A -infected pregnant sows, that it significantly improves the reproductive performance of sows and reduces the incidence of bloat in sows and diarrhea in newborn piglets. Significantly increased the abundance of intestinal probiotics ( Bacillus subtilis , Lactobacillus , Limosilactobacillus reuteri , Lactobacillus johnsonii , Muribaculaceae , Lactobacillus amylovorus , and Lactobacillus reuteri ) in sows and decreased the relative abundance of Clostridium perfringens type A after feeding Bacillus subtilis . These probiotics can repair the intestinal tissue histomorphology and structure and enhance the expression of MUC2 and sIgA in sows, further strengthening the mucosal immune function of the intestine. Bacillus subtilis can also reduce inflammatory factors(CRP, IL-1β, and IFN-ɤ) levels and the inflammatory response in sows and newborn piglets. Taken together, our results suggest that dietary supplementation with Bacillus subtilis PB6 could benefit the sow bloat and piglets diarrhea, and improve intestinal barrier function, and microbial balance of sows. Sow Clostridium perfringens type A Bacillus subtilis PB6 Piglet Figures Figure 1 Figure 2 Figure 3 Figure 4 Full Text Additional Declarations No competing interests reported. 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. 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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-7359451","acceptedTermsAndConditions":true,"allowDirectSubmit":true,"archivedVersions":[],"articleType":"Research Article","associatedPublications":[],"authors":[{"id":507809964,"identity":"a9c51664-60b7-4507-8d5d-65552b1e2062","order_by":0,"name":"Mengran Zhang","email":"","orcid":"","institution":"Northwest Agriculture and F University","correspondingAuthor":false,"prefix":"","firstName":"Mengran","middleName":"","lastName":"Zhang","suffix":""},{"id":507809966,"identity":"aabd4501-29d9-4818-bf17-bc129808ce33","order_by":1,"name":"Aohang-Yu Yu","email":"","orcid":"","institution":"Northwest Agriculture and F University","correspondingAuthor":false,"prefix":"","firstName":"Aohang-Yu","middleName":"","lastName":"Yu","suffix":""},{"id":507809968,"identity":"e781451c-6832-41db-bc11-f4052ec08569","order_by":2,"name":"Chaochi Wang","email":"","orcid":"","institution":"Northwest Agriculture and F University","correspondingAuthor":false,"prefix":"","firstName":"Chaochi","middleName":"","lastName":"Wang","suffix":""},{"id":507809970,"identity":"aa553f5e-c852-4a29-87cd-335a64909d60","order_by":3,"name":"Jiayue Bai","email":"","orcid":"","institution":"Northwest Agriculture and F University","correspondingAuthor":false,"prefix":"","firstName":"Jiayue","middleName":"","lastName":"Bai","suffix":""},{"id":507809971,"identity":"b6f06e8f-1ddf-44fb-8a66-31e9570df6e8","order_by":4,"name":"Chaojie Chen","email":"","orcid":"","institution":"Northwest Agriculture and F University","correspondingAuthor":false,"prefix":"","firstName":"Chaojie","middleName":"","lastName":"Chen","suffix":""},{"id":507809972,"identity":"53645253-6610-47c0-a13e-92fbb0bf60dc","order_by":5,"name":"Chenchen Wu","email":"data:image/png;base64,iVBORw0KGgoAAAANSUhEUgAAAZAAAAAyAQMAAABI0h/eAAAABlBMVEX///8AAABVwtN+AAAACXBIWXMAAA7EAAAOxAGVKw4bAAAAyElEQVRIiWNgGAWjYNCCin9y/OzNBw58+EG0ljMHjCV7jiUenNlDrA7GtgOJG27kGB/mYCNCtcGN5IePC9vuAG058+EwAw+DPL/YAUJa0oyNZ5x7BvRL74bDBRYMhjNnJ+DXYnYjh02ap4wZaMvZDYdn8DAkGNwmrIX9Nw8bM8gvDw7zsBGnhY2Zp+0wSAsDcVrszzwzluY5kwYKZANgIEsQ9otke/LDzzwVNqCofPzhww8beX5pAlrQgQRpykfBKBgFo2AUYAcAPDRLv3ZVmsgAAAAASUVORK5CYII=","orcid":"","institution":"Northwest Agriculture and F University","correspondingAuthor":true,"prefix":"","firstName":"Chenchen","middleName":"","lastName":"Wu","suffix":""}],"badges":[],"createdAt":"2025-08-12 23:23:13","currentVersionCode":1,"declarations":"","doi":"10.21203/rs.3.rs-7359451/v1","doiUrl":"https://doi.org/10.21203/rs.3.rs-7359451/v1","draftVersion":[],"editorialEvents":[],"editorialNote":"","failedWorkflow":false,"files":[{"id":90538974,"identity":"a31fc29d-1874-413b-bdca-9e305c413da3","added_by":"auto","created_at":"2025-09-03 22:06:00","extension":"jpg","order_by":1,"title":"Figure 1","display":"","copyAsset":false,"role":"figure","size":824627,"visible":true,"origin":"","legend":"\u003cp\u003e\u003cstrong\u003eEffect of feeding \u003c/strong\u003e\u003cem\u003e\u003cstrong\u003eBacillus subtilis \u003c/strong\u003e\u003c/em\u003e\u003cstrong\u003eon reproductive performance of pregnancy sows. A\u003c/strong\u003e, \u003cem\u003eClostridium perfringens \u003c/em\u003epollution rate of sows of pregnancy 0-30d, 30-60d, 60-90d, 90-110d. Black color represents heavy pollution, and brown color represents moderate pollution. \u003cstrong\u003eB\u003c/strong\u003e, Sow bloat rate before BS 1 month, after BS 1 month and BS 2 months; Diarrhea of piglets in control and BS group. \u003cstrong\u003eC\u003c/strong\u003e, The number of births, the number of live births, born litter of weight,weaning litter weight in control and BS group. \u003cstrong\u003eD\u003c/strong\u003e, \u003cem\u003eClostridium perfringens type A \u003c/em\u003eexpression and a-toxin level in control and BS group.E,Average daily feed intake of piglets and feed conversion ration in control and BS group.\u003c/p\u003e","description":"","filename":"Figure1.jpg","url":"https://assets-eu.researchsquare.com/files/rs-7359451/v1/760aa5e1f249502f5c23c945.jpg"},{"id":90539110,"identity":"58abc7f6-c220-4300-b860-ddbaec9235ff","added_by":"auto","created_at":"2025-09-03 22:14:00","extension":"jpg","order_by":2,"title":"Figure 2","display":"","copyAsset":false,"role":"figure","size":2288250,"visible":true,"origin":"","legend":"\u003cp\u003e\u003cstrong\u003eEffect of feeding \u003c/strong\u003e\u003cem\u003e\u003cstrong\u003eBacillus subtilis \u003c/strong\u003e\u003c/em\u003e\u003cstrong\u003eon diversity of the intestinal flora in sow and newborn piglets. A\u003c/strong\u003e, The same microbiological flora of sows between control and BS group; \u003cstrong\u003eB\u003c/strong\u003e, The same microbiological flora between control and BS group at birth 7d and weaned piglets. \u003cstrong\u003eC\u003c/strong\u003e, Kruskal-Walls H test for sobs index of control and BS group in sows, birth 7d, and weaned piglets. \u003cstrong\u003eD\u003c/strong\u003e, Relative abundance on Phylum level in community barplot analysis in sows, birth 7d and weaned piglets of control and BS group; Firmicutes/Bacteroidota ratio in sows, birth 7d and weaned piglets of control and BS group. \u003cstrong\u003eE\u003c/strong\u003e, Relative abundance on the Family level in community barplot analysis in sows, birth 7d and weaned piglets of control and BS group; Clostridiaceae ratio in sows, birth 7d and weaned piglets of control and BS group. \u003cstrong\u003eF, \u003c/strong\u003eCommunity heatmap analysis on Genus level of sows in BS and control group. \u003cstrong\u003eG\u003c/strong\u003e,Community heatmap analysis on Genus level of piglets at birth 7d and weaned piglets of control and BS group; \u003cstrong\u003eH\u003c/strong\u003e, Relative abundance of \u003cem\u003eLactobacillus\u003c/em\u003e, \u003cem\u003eLimosilactobacillus reuteri\u003c/em\u003e, \u003cem\u003eLactobacillus johnsonii\u003c/em\u003e, \u003cem\u003eMuribaculaceae\u003c/em\u003e, \u003cem\u003eLactobacillus amylovorus\u003c/em\u003e, \u003cem\u003eLactobacillus reuteri \u003c/em\u003ein sows of control and BS group. \u003cstrong\u003eI\u003c/strong\u003e, Relative abundance of \u003cem\u003eEscherichia-shigella\u003c/em\u003e, \u003cem\u003eLigilactobacillus, \u003c/em\u003eand \u003cem\u003eIntestinimonas \u003c/em\u003ein birth 7d and weaned piglets of control and BS group.\u003c/p\u003e","description":"","filename":"Figure21.jpg","url":"https://assets-eu.researchsquare.com/files/rs-7359451/v1/d8dc3b5579ee188c5cd84b44.jpg"},{"id":90539252,"identity":"3a8d6c9d-de49-4775-897c-75f287472672","added_by":"auto","created_at":"2025-09-03 22:22:00","extension":"jpg","order_by":3,"title":"Figure 3","display":"","copyAsset":false,"role":"figure","size":4295153,"visible":true,"origin":"","legend":"\u003cp\u003e\u003cstrong\u003eEffect of feeding \u003c/strong\u003e\u003cem\u003e\u003cstrong\u003eBacillus subtilis \u003c/strong\u003e\u003c/em\u003e\u003cstrong\u003eon histomorphology of the sow's organs. A B C, D\u003c/strong\u003e, Anatomy of the intestinal tract of the sow. \u003cstrong\u003eE \u003c/strong\u003eThe histomorphology of heart, liver, lung, spleen, and kidney after \u003cem\u003eClostridium perfringens type A \u003c/em\u003einfection sows in the control group (Feeding normal diets) and BS group (adding \u003cem\u003e\u003cstrong\u003eBacillus subtilis \u003c/strong\u003e\u003c/em\u003eto diets). \u003cstrong\u003eF \u003c/strong\u003eThe histomorphology of intestine after \u003cem\u003eClostridium perfringens type A \u003c/em\u003einfection sow in the control group (Feeding normal diets) and BS group (adding \u003cem\u003e\u003cstrong\u003eBacillus subtilis \u003c/strong\u003e\u003c/em\u003eto diets), top two lines were stained H\u0026amp;E, bottom two lines were stained A\u0026amp;B (×200). \u003cstrong\u003eG\u003c/strong\u003e, The MUC2 and sIgA (brown color) distribution in intestinal were analyzed by immunohistochemistry after \u003cem\u003eClostridium perfringens type A \u003c/em\u003einfection sow in the control group (Feeding normal diets) and BS group (adding \u003cem\u003e\u003cstrong\u003eBacillus subtilis \u003c/strong\u003e\u003c/em\u003eto diets)(×200).\u003cstrong\u003eH, \u003c/strong\u003eThe MUC2 and sIgA (brown color) levels in the intestine were analyzed by ELISA after \u003cem\u003eClostridium perfringens type A \u003c/em\u003einfection sows in the control group (Feeding normal diets) and BS group(adding \u003cem\u003e\u003cstrong\u003eBacillus subtilis \u003c/strong\u003e\u003c/em\u003eto diets).\u003c/p\u003e","description":"","filename":"Figure31.jpg","url":"https://assets-eu.researchsquare.com/files/rs-7359451/v1/0d820543f53b860dd8ca7892.jpg"},{"id":90539113,"identity":"6419e2f6-e83f-4557-86d3-b38a31299f54","added_by":"auto","created_at":"2025-09-03 22:14:00","extension":"jpg","order_by":4,"title":"Figure 4","display":"","copyAsset":false,"role":"figure","size":712133,"visible":true,"origin":"","legend":"\u003cp\u003e\u003cstrong\u003eEffect of feeding \u003c/strong\u003e\u003cem\u003e\u003cstrong\u003eBacillus subtilis \u003c/strong\u003e\u003c/em\u003e\u003cstrong\u003eon pro-inflammatory and anti-inflammatory factors in sow and newborn piglets.\u003c/strong\u003e\u003c/p\u003e","description":"","filename":"Figure4.jpg","url":"https://assets-eu.researchsquare.com/files/rs-7359451/v1/aa14441ac314d13e21b62c5c.jpg"},{"id":104494092,"identity":"95953c94-de51-4f0b-8829-8940fe344089","added_by":"auto","created_at":"2026-03-12 12:12:49","extension":"pdf","order_by":1,"title":"","display":"","copyAsset":false,"role":"manuscript-pdf","size":8697890,"visible":true,"origin":"","legend":"","description":"","filename":"manuscript2025821.pdf","url":"https://assets-eu.researchsquare.com/files/rs-7359451/v1_covered_8a52b7d8-a7e6-48d5-80c3-d349081a98c5.pdf"}],"financialInterests":"No competing interests reported.","formattedTitle":"\u003cp\u003eEffect of feeding Bacillus subtilis on intestinal bloat and microbiological structure in sows infected with Clostridium perfringens type A\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":"Sow, Clostridium perfringens type A, Bacillus subtilis, PB6, Piglet","lastPublishedDoi":"10.21203/rs.3.rs-7359451/v1","lastPublishedDoiUrl":"https://doi.org/10.21203/rs.3.rs-7359451/v1","license":{"name":"CC BY 4.0","url":"https://creativecommons.org/licenses/by/4.0/"},"manuscriptAbstract":"\u003cp\u003e\u003cem\u003eBacillus subtilis \u003c/em\u003eis aerobic or facultatively anaerobic.After entering the gastrointestinal tract, spores colonize the gut, inhibiting the growth of harmful aerobic bacteria (\u003cem\u003eEscherichia coli\u003c/em\u003e, \u003cem\u003eStreptococcus\u003c/em\u003e, \u003cem\u003eStaphylococcus aureus\u003c/em\u003e).However, it is not clear whether \u003cem\u003eBacillus subtilis \u003c/em\u003ecan inhibit \u003cem\u003eClostridium perfringens type A \u003c/em\u003einfection. In this study, \u003cem\u003eBacillus subtilis \u003c/em\u003ePB6 was added to the diets of \u003cem\u003eClostridium perfringens type A\u003c/em\u003e-infected pregnant sows, that it significantly improves the reproductive performance of sows and reduces the incidence of bloat in sows and diarrhea in newborn piglets. Significantly increased the abundance of intestinal probiotics (\u003cem\u003eBacillus subtilis\u003c/em\u003e, \u003cem\u003eLactobacillus\u003c/em\u003e, \u003cem\u003eLimosilactobacillus reuteri\u003c/em\u003e, \u003cem\u003eLactobacillus johnsonii\u003c/em\u003e, \u003cem\u003eMuribaculaceae\u003c/em\u003e, \u003cem\u003eLactobacillus amylovorus\u003c/em\u003e, and \u003cem\u003eLactobacillus reuteri\u003c/em\u003e) in sows and decreased the relative abundance of \u003cem\u003eClostridium perfringens type A \u003c/em\u003eafter feeding \u003cem\u003eBacillus subtilis\u003c/em\u003e. These probiotics can repair the intestinal tissue histomorphology and structure and enhance the expression of MUC2 and sIgA in sows, further strengthening the mucosal immune function of the intestine. \u003cem\u003eBacillus subtilis \u003c/em\u003ecan also reduce inflammatory factors(CRP, IL-1β, and IFN-ɤ) levels and the inflammatory response in sows and newborn piglets. Taken together, our results suggest that dietary supplementation with \u003cem\u003eBacillus subtilis \u003c/em\u003ePB6 could benefit the sow bloat and piglets diarrhea, and improve intestinal barrier function, and microbial balance of sows.\u003c/p\u003e","manuscriptTitle":"Effect of feeding Bacillus subtilis on intestinal bloat and microbiological structure in sows infected with Clostridium perfringens type A","msid":"","msnumber":"","nonDraftVersions":[{"code":1,"date":"2025-09-03 22:05:55","doi":"10.21203/rs.3.rs-7359451/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":"7056b278-e8de-4c5d-b5e4-b8e5882e2d72","owner":[],"postedDate":"September 3rd, 2025","published":true,"recentEditorialEvents":[],"rejectedJournal":[],"revision":"","amendment":"","status":"posted","subjectAreas":[],"tags":[],"updatedAt":"2026-03-12T12:12:24+00:00","versionOfRecord":[],"versionCreatedAt":"2025-09-03 22:05:55","video":"","vorDoi":"","vorDoiUrl":"","workflowStages":[]},"version":"v1","identity":"rs-7359451","journalConfig":"researchsquare"},"__N_SSP":true},"page":"/article/[identity]/[[...version]]","query":{"redirect":"/article/rs-7359451","identity":"rs-7359451","version":["v1"]},"buildId":"8U1c8b4HqxoKbykW_rLl7","isFallback":false,"isExperimentalCompile":false,"dynamicIds":[84888],"gssp":true,"scriptLoader":[]}