From wild to farm: gut bacteriome differences and probiotic potential of Pantoea agglomerans in two-spotted cricket (Gryllus bimaculatus) rearing

From wild to farm: gut bacteriome differences and probiotic potential of Pantoea agglomerans in two-spotted cricket (Gryllus bimaculatus) rearing | 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 From wild to farm: gut bacteriome differences and probiotic potential of Pantoea agglomerans in two-spotted cricket (Gryllus bimaculatus) rearing Diego Cruz, Zaki Saati-Santamaria, Luisa Achury-Arrubla, Paula Garcia-Fraile This is a preprint; it has not been peer reviewed by a journal. https://doi.org/ 10.21203/rs.3.rs-8758672/v1 This work is licensed under a CC BY 4.0 License Status: Published Journal Publication published 22 Apr, 2026 Read the published version in Probiotics and Antimicrobial Proteins → Version 1 posted 12 You are reading this latest preprint version Abstract The gut microbiome plays a crucial role in insect nutrition and performance, yet its targeted exploitation in cricket farming remains underexplored. Here, we combined gut microbiota profiling of wild and farmed Gryllus bimaculatus with probiotic testing of host-derived bacterial isolates to explore microbiome-informed strategies for sustainable cricket farming. Wild crickets exhibited higher Shannon diversity but lower phylogenetic diversity than farmed counterparts. Wild populations were enriched in Oscillospiraceae and Christensenellaceae families, while farmed crickets showed higher abundance of Parabacteroides . From 199 bacterial isolates, wild populations showed higher frequencies of uricolytic capabilities (44% vs 31%), related to nitrogen recycling, while farmed crickets had more pectinolytic isolates (70% vs 50%), linked to plant fiber degradation. Pantoea agglomerans I53BLB, which demonstrated broad enzymatic capabilities, was selected for probiotic evaluation; we further provide its genome sequence and analysis to contextualize its metabolic and probiotic potential. A feeding experiment compared control chicken feed versus a high-fiber diet formulated with agricultural by-products, each supplemented with water, live or heat-inactivated P. agglomerans . A significant diet × probiotic interaction was observed for weight gain and adult emergence. Live P. agglomerans enhanced performance (weight and adult emergence) only when combined with the high-fiber, suggesting a symbiotic effect likely related with carbohydrate digestion. No effects were observed on survival or reproductive output. Notably, the high-fiber diet alone performed comparably to commercial feed, suggesting potential for sustainable cricket production using agricultural by-products. These findings demonstrate the feasibility of microbiome informed probiotic strategies to enhance cricket farming efficiency while reducing feed costs. Edible insects Gryllus bimaculatus Gut microbiota Probiotics Genomics Pantoea agglomerans Full Text Additional Declarations No competing interests reported. Cite Share Download PDF Status: Published Journal Publication published 22 Apr, 2026 Read the published version in Probiotics and Antimicrobial Proteins → Version 1 posted Editorial decision: Revision requested 25 Feb, 2026 Reviews received at journal 24 Feb, 2026 Reviews received at journal 24 Feb, 2026 Reviews received at journal 23 Feb, 2026 Reviewers agreed at journal 05 Feb, 2026 Reviewers agreed at journal 04 Feb, 2026 Reviewers agreed at journal 03 Feb, 2026 Reviewers agreed at journal 03 Feb, 2026 Reviewers invited by journal 03 Feb, 2026 Editor assigned by journal 02 Feb, 2026 Submission checks completed at journal 02 Feb, 2026 First submitted to journal 01 Feb, 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-8758672","acceptedTermsAndConditions":true,"allowDirectSubmit":false,"archivedVersions":[],"articleType":"Research Article","associatedPublications":[],"authors":[{"id":585982561,"identity":"5c56dbbf-c7b3-405b-a782-6a5770dbd5ac","order_by":0,"name":"Diego Cruz","email":"data:image/png;base64,iVBORw0KGgoAAAANSUhEUgAAAZAAAAAyAQMAAABI0h/eAAAABlBMVEX///8AAABVwtN+AAAACXBIWXMAAA7EAAAOxAGVKw4bAAAA50lEQVRIie2RsQrCMBCGI4FMabsmCD5DoCCK0r7KScBJn0BBJ0dd+xhOzoWAXYSudRRBHBw6lU7iFcTRdBTMN/wQuI/7jxDicPwipolxEzTFGLRVpk0wwBAtlPSjcNVO8TNq7iVE0T47VbJeChIE6XdFGjYdJqD1/jQ/dPlREJnAd0UZ3g+9mmqVokKYIKqwFFMmqEIOK63yx03WT0Hi3FIMt9ArBxOpYsaEt8EtxFIMb+l3EshAFrdw5G0FF4VF8XNzLUtYxH6uL+e6GveCnaUYwpq/mKzfL26dR2iJEbeZdDgcjj/lBT4QQ4ZaQSQVAAAAAElFTkSuQmCC","orcid":"","institution":"Universidad de Salamanca","correspondingAuthor":true,"prefix":"","firstName":"Diego","middleName":"","lastName":"Cruz","suffix":""},{"id":585982562,"identity":"a79bd85f-71d8-417a-90df-de371a80e1cb","order_by":1,"name":"Zaki Saati-Santamaria","email":"","orcid":"","institution":"Universidad de Salamanca","correspondingAuthor":false,"prefix":"","firstName":"Zaki","middleName":"","lastName":"Saati-Santamaria","suffix":""},{"id":585982563,"identity":"3da08640-d063-4751-a624-bebf01c55105","order_by":2,"name":"Luisa Achury-Arrubla","email":"","orcid":"","institution":"Universidad de Salamanca","correspondingAuthor":false,"prefix":"","firstName":"Luisa","middleName":"","lastName":"Achury-Arrubla","suffix":""},{"id":585982564,"identity":"5e3ff7c5-4961-4f10-b270-7e2f69b27ae5","order_by":3,"name":"Paula Garcia-Fraile","email":"","orcid":"","institution":"Universidad de Salamanca","correspondingAuthor":false,"prefix":"","firstName":"Paula","middleName":"","lastName":"Garcia-Fraile","suffix":""}],"badges":[],"createdAt":"2026-02-01 20:23:15","currentVersionCode":1,"declarations":"","doi":"10.21203/rs.3.rs-8758672/v1","doiUrl":"https://doi.org/10.21203/rs.3.rs-8758672/v1","draftVersion":[],"editorialEvents":[{"content":"https://doi.org/10.1007/s12602-026-11001-1","type":"published","date":"2026-04-22T15:58:40+00:00"}],"editorialNote":"","failedWorkflow":false,"files":[{"id":107929310,"identity":"67a232a5-7b24-4778-8ee4-a997c7eeda5d","added_by":"auto","created_at":"2026-04-27 16:14:48","extension":"pdf","order_by":1,"title":"","display":"","copyAsset":false,"role":"manuscript-pdf","size":1214903,"visible":true,"origin":"","legend":"","description":"","filename":"Fromwildtofarmgutbacteriomeintwospottedcricket.pdf","url":"https://assets-eu.researchsquare.com/files/rs-8758672/v1_covered_32fd98a4-1ce6-4950-9097-f3ed11c651e1.pdf"}],"financialInterests":"No competing interests reported.","formattedTitle":"From wild to farm: gut bacteriome differences and probiotic potential of Pantoea agglomerans in two-spotted cricket (Gryllus bimaculatus) rearing","fulltext":[],"fulltextSource":"","fullText":"","funders":[],"hasAdminPriorityOnWorkflow":false,"hasManuscriptDocX":false,"hasOptedInToPreprint":true,"hasPassedJournalQc":"","hasAnyPriority":false,"hideJournal":false,"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":"probiotics-and-antimicrobial-proteins","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":false,"externalIdentity":"paap","sideBox":"Learn more about [Probiotics and Antimicrobial Proteins](http://link.springer.com/journal/12601)","snPcode":"12602","submissionUrl":"https://submission.nature.com/new-submission/12602/3","title":"Probiotics and Antimicrobial Proteins","twitterHandle":"","acdcEnabled":true,"dfaEnabled":true,"editorialSystem":"em","reportingPortfolio":"Springer Hybrid","inReviewEnabled":true,"inReviewRevisionsEnabled":false},"keywords":"Edible insects, Gryllus bimaculatus, Gut microbiota, Probiotics, Genomics, Pantoea agglomerans","lastPublishedDoi":"10.21203/rs.3.rs-8758672/v1","lastPublishedDoiUrl":"https://doi.org/10.21203/rs.3.rs-8758672/v1","license":{"name":"CC BY 4.0","url":"https://creativecommons.org/licenses/by/4.0/"},"manuscriptAbstract":"\u003cp\u003eThe gut microbiome plays a crucial role in insect nutrition and performance, yet its targeted exploitation in cricket farming remains underexplored. Here, we combined gut microbiota profiling of wild and farmed \u003cem\u003eGryllus bimaculatus\u003c/em\u003e with probiotic testing of host-derived bacterial isolates to explore microbiome-informed strategies for sustainable cricket farming. Wild crickets exhibited higher Shannon diversity but lower phylogenetic diversity than farmed counterparts. Wild populations were enriched in \u003cem\u003eOscillospiraceae\u003c/em\u003e and \u003cem\u003eChristensenellaceae\u003c/em\u003e families, while farmed crickets showed higher abundance of \u003cem\u003eParabacteroides\u003c/em\u003e. From 199 bacterial isolates, wild populations showed higher frequencies of uricolytic capabilities (44% vs 31%), related to nitrogen recycling, while farmed crickets had more pectinolytic isolates (70% vs 50%), linked to plant fiber degradation. \u003cem\u003ePantoea agglomerans\u003c/em\u003e I53BLB, which demonstrated broad enzymatic capabilities, was selected for probiotic evaluation; we further provide its genome sequence and analysis to contextualize its metabolic and probiotic potential. A feeding experiment compared control chicken feed versus a high-fiber diet formulated with agricultural by-products, each supplemented with water, live or heat-inactivated \u003cem\u003eP. agglomerans\u003c/em\u003e. A significant diet \u0026times; probiotic interaction was observed for weight gain and adult emergence. 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