Accelerating Gut Microbiome Research with Robust Sample Collection

Accelerating Gut Microbiome Research with Robust Sample Collection | 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 Methodology Accelerating Gut Microbiome Research with Robust Sample Collection Zoe J. Zreloff, Danielle Lange, Suzanne D. Vernon, Martha R. Carlin, and 1 more This is a preprint; it has not been peer reviewed by a journal. https://doi.org/ 10.21203/rs.3.rs-129091/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 Background. Inferior quality of biological material compromises data, slows discovery, and wastes research funds. The gut microbiome plays a critical role in human health and disease, yet little attention has been given to optimizing collection and processing methods of human stool. Methods. We collected the entire bowel movement from 2 healthy volunteers: one to examine stool sample heterogeneity and one to test stool sample handling parameters. Sequencing and bioinformatic analyses were used to examine the microbiome composition. Results. The microbiome profile varied depending on where the subsample was obtained from the stool. The exterior cortex of the stool was rich in specific phyla and deficient in others while the interior core of the stool revealed opposite microbiome profiles. Sample processing also resulted in varying microbiome profiles. Homogenization and stabilization at 4°C gave superior microbial diversity profiles compared to the fresh or frozen subsamples of the same stool sample. Bacterial proliferation continued in the fresh subsample when processed at ambient temperature. Bacteroidetes proliferated and Firmicutes diminished during the 30-minute processing of fresh sample. The frozen sample had good overall diversity but Proteobacteria diminished likely because of the freeze/thaw. Conclusions. The microbiome profile is specific to the section of the stool being sampled. Stool sample collection, homogenization, and stabilization at 4°C for 24 hours provides a “neat”, high-quality sample of sufficient quantity that can be banked into aliquots with nearly identical microbial diversity profiles. This collection pipeline is essential to accelerate our understanding of the gut microbiome in health and disease General Microbiology Stool Fecal Microbiome Microbiota Heterogeneous Homogeneous Sample Subsample Figures Figure 1 Figure 1 Figure 2 Figure 2 Figure 3 Figure 3 Figure 4 Figure 4 Figure 5 Figure 5 Figure 6 Figure 6 Figure 7 Figure 7 Full Text Due to technical limitations, full-text HTML conversion of this manuscript could not be completed. However, the latest manuscript can be downloaded and accessed as a PDF. Tables Table 1. Wilkinson Rank Sum tests of alpha diversity metrics for the various sample processing protocols Wilcoxon Rank Sum Test Shannon Simpson Cohorts Statistic P-value* Statistic P-value* Fresh ↔ 4C -1.9845 0.0472 -2.6112 0.009 Fresh ↔ Frozen 1.3578 0.1745 2.6112 0.009 4C ↔ Frozen 2.6112 0.009 2.6112 0.009 PERMANOVA Analysis Cohorts Statistic P-value* Fresh ↔ 4C 1.97 0.043 Fresh ↔ Frozen 4.433 0.006 4C ↔ Frozen 2.567 0.005 Supplementary Files TableS1MetadataTableofDada2results.xlsx FigureS1QCplots.pdf 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. 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-129091","acceptedTermsAndConditions":true,"allowDirectSubmit":true,"archivedVersions":[],"articleType":"Methodology","associatedPublications":[],"authors":[{"id":6506467,"identity":"badb8032-d34f-4010-9cb3-f4bfbcb237ff","order_by":0,"name":"Zoe J. Zreloff","email":"","orcid":"","institution":"Teh BioCollective, LLC","correspondingAuthor":false,"prefix":"","firstName":"Zoe","middleName":"J.","lastName":"Zreloff","suffix":""},{"id":6506468,"identity":"0c5e32b0-aa3e-4815-95ab-40dac3ffe6ce","order_by":1,"name":"Danielle Lange","email":"","orcid":"","institution":"The BioCollective, LLC","correspondingAuthor":false,"prefix":"","firstName":"Danielle","middleName":"","lastName":"Lange","suffix":""},{"id":6506469,"identity":"34c6f402-1dab-4c9f-ad54-5e3d51b5f232","order_by":2,"name":"Suzanne D. Vernon","email":"","orcid":"","institution":"The BioCollective, LLC","correspondingAuthor":false,"prefix":"","firstName":"Suzanne","middleName":"D.","lastName":"Vernon","suffix":""},{"id":6506470,"identity":"29fa80ed-2ad3-4b72-b6be-dabb863f0d5c","order_by":3,"name":"Martha R. Carlin","email":"","orcid":"","institution":"The BioCollective, LLC","correspondingAuthor":false,"prefix":"","firstName":"Martha","middleName":"R.","lastName":"Carlin","suffix":""},{"id":6506471,"identity":"2358fde1-fb0a-4fea-a3fc-dcb70884a7af","order_by":4,"name":"Raul de Jesus Cano","email":"data:image/png;base64,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","orcid":"https://orcid.org/0000-0001-6888-5018","institution":"The BioCollective, LLC","correspondingAuthor":true,"prefix":"","firstName":"Raul","middleName":"de Jesus","lastName":"Cano","suffix":""}],"badges":[],"createdAt":"2020-12-15 13:16:49","currentVersionCode":1,"declarations":"","doi":"10.21203/rs.3.rs-129091/v1","doiUrl":"https://doi.org/10.21203/rs.3.rs-129091/v1","draftVersion":[],"editorialEvents":[],"editorialNote":"","failedWorkflow":false,"files":[{"id":4779727,"identity":"c084b63a-92a3-4632-ae2f-6a8ec4f894f7","added_by":"auto","created_at":"2021-01-07 15:08:33","extension":"png","order_by":1,"title":"Figure 1","display":"","copyAsset":false,"role":"figure","size":76051,"visible":true,"origin":"","legend":"Schematic of stool dissection. The stool sample from one volunteer was cut into \nsections and analyzed to determine differences in microbiome composition. \n ","description":"","filename":"1.png","url":"https://assets-eu.researchsquare.com/files/rs-129091/v1/841e874b35d977bb3bd8c69e.png"},{"id":4338936,"identity":"20b1e613-2603-4057-91da-1060a8fa7d4f","added_by":"auto","created_at":"2020-12-17 16:42:39","extension":"png","order_by":1,"title":"Figure 1","display":"","copyAsset":false,"role":"figure","size":76051,"visible":true,"origin":"","legend":"Schematic of stool dissection. The stool sample from one volunteer was cut into \nsections and analyzed to determine differences in microbiome composition. \n ","description":"","filename":"1.png","url":"https://assets-eu.researchsquare.com/files/rs-129091/v1/8b479c2ad79f937074054cd3.png"},{"id":4779726,"identity":"a042244b-9ef1-4d0c-82b2-891d28fa3848","added_by":"auto","created_at":"2021-01-07 15:08:33","extension":"png","order_by":2,"title":"Figure 2","display":"","copyAsset":false,"role":"figure","size":51360,"visible":true,"origin":"","legend":"Principal Coordinates Analysis plot based on Bray-Curtis dissimilarity matrix for all stool sub-samples. The PCoA plot shows distinct clustering of the stool samples based on the region or segment of the stool from which they originated. The legend Indicates the 6 different sampling sites, each one cm apart. The plot illustrates the coordinates for each individual sample. ","description":"","filename":"2.png","url":"https://assets-eu.researchsquare.com/files/rs-129091/v1/3b64722a75268afa6322262f.png"},{"id":4338959,"identity":"b2de70df-24b2-4af7-8dfb-5ba9e3f098b9","added_by":"auto","created_at":"2020-12-17 16:42:39","extension":"png","order_by":2,"title":"Figure 2","display":"","copyAsset":false,"role":"figure","size":51360,"visible":true,"origin":"","legend":"Principal Coordinates Analysis plot based on Bray-Curtis dissimilarity matrix for all stool sub-samples. The PCoA plot shows distinct clustering of the stool samples based on the region or segment of the stool from which they originated. The legend Indicates the 6 different sampling sites, each one cm apart. The plot illustrates the coordinates for each individual sample. ","description":"","filename":"2.png","url":"https://assets-eu.researchsquare.com/files/rs-129091/v1/128d944b21547e0b42edf581.png"},{"id":4779729,"identity":"9484ba72-ee4f-43a9-b208-24d0d30d1f0f","added_by":"auto","created_at":"2021-01-07 15:08:34","extension":"png","order_by":3,"title":"Figure 3","display":"","copyAsset":false,"role":"figure","size":147450,"visible":true,"origin":"","legend":" Differential abundance of taxa in dissected stool. The figure summarizes the \ndistribution of 12 different genera present in high, medium and low relative abundance represented in each section of the stool. ","description":"","filename":"3.png","url":"https://assets-eu.researchsquare.com/files/rs-129091/v1/422ec1f8c415b2c2accbe061.png"},{"id":4338961,"identity":"5ba8d253-6a74-45d4-bafd-05c4d4e42d7c","added_by":"auto","created_at":"2020-12-17 16:42:39","extension":"png","order_by":3,"title":"Figure 3","display":"","copyAsset":false,"role":"figure","size":147450,"visible":true,"origin":"","legend":" Differential abundance of taxa in dissected stool. The figure summarizes the \ndistribution of 12 different genera present in high, medium and low relative abundance represented in each section of the stool. ","description":"","filename":"3.png","url":"https://assets-eu.researchsquare.com/files/rs-129091/v1/b6b54152c435c4162c9c4201.png"},{"id":4779728,"identity":"f3cdc0e3-839b-44c4-9544-6483092a03ed","added_by":"auto","created_at":"2021-01-07 15:08:33","extension":"png","order_by":4,"title":"Figure 4","display":"","copyAsset":false,"role":"figure","size":73122,"visible":true,"origin":"","legend":"Sample Processing Workflow. This figure shows the steps taken in sample collection and processing for comparing sample preparation protocols. \n ","description":"","filename":"4.png","url":"https://assets-eu.researchsquare.com/files/rs-129091/v1/b13b495439b454e89cc592e3.png"},{"id":4338960,"identity":"c12e9b83-7012-483a-9dd5-493f99583ae1","added_by":"auto","created_at":"2020-12-17 16:42:39","extension":"png","order_by":4,"title":"Figure 4","display":"","copyAsset":false,"role":"figure","size":73122,"visible":true,"origin":"","legend":"Sample Processing Workflow. This figure shows the steps taken in sample collection and processing for comparing sample preparation protocols. \n ","description":"","filename":"4.png","url":"https://assets-eu.researchsquare.com/files/rs-129091/v1/73250477533d0f1785a98127.png"},{"id":4779629,"identity":"6a551947-d3be-4203-8d78-8f1eb741d98b","added_by":"auto","created_at":"2021-01-07 15:05:33","extension":"png","order_by":5,"title":"Figure 5","display":"","copyAsset":false,"role":"figure","size":50238,"visible":true,"origin":"","legend":"Taxonomic bar graph at the Phylum level for aliquots analyzed from three different sample processing methods. Relative abundance as a percent of total phyla was plotted as a stacked bar graph to illustrate the variations in relative abundance of individual phyla based on sample processing protocol and sampling time. ","description":"","filename":"5.png","url":"https://assets-eu.researchsquare.com/files/rs-129091/v1/4a2b310441010f88b29ee570.png"},{"id":4338963,"identity":"72d39965-9843-46b2-9725-f5755f94f074","added_by":"auto","created_at":"2020-12-17 16:42:39","extension":"png","order_by":5,"title":"Figure 5","display":"","copyAsset":false,"role":"figure","size":50238,"visible":true,"origin":"","legend":"Taxonomic bar graph at the Phylum level for aliquots analyzed from three different sample processing methods. Relative abundance as a percent of total phyla was plotted as a stacked bar graph to illustrate the variations in relative abundance of individual phyla based on sample processing protocol and sampling time. ","description":"","filename":"5.png","url":"https://assets-eu.researchsquare.com/files/rs-129091/v1/51510543e4065e87abebf011.png"},{"id":4779730,"identity":"b091dc51-9fdf-425f-a73d-1700744b3860","added_by":"auto","created_at":"2021-01-07 15:08:34","extension":"png","order_by":6,"title":"Figure 6","display":"","copyAsset":false,"role":"figure","size":49173,"visible":true,"origin":"","legend":"Firmicutes to Bacteroidetes ratio for aliquots analyzed from three different sample processing protocols. The ratio of Firmicutes:Bacteroidetes (F:B) was plotted as a side-by-side bar graph to illustrate the variations in F:B ratio for each sample processing protocol and sampling time. \n ","description":"","filename":"6.png","url":"https://assets-eu.researchsquare.com/files/rs-129091/v1/6f61579ce039c681cf51d3b2.png"},{"id":4338962,"identity":"82f98b2d-e839-4b3b-8d5f-0b25313184ac","added_by":"auto","created_at":"2020-12-17 16:42:39","extension":"png","order_by":6,"title":"Figure 6","display":"","copyAsset":false,"role":"figure","size":49173,"visible":true,"origin":"","legend":"Firmicutes to Bacteroidetes ratio for aliquots analyzed from three different sample processing protocols. The ratio of Firmicutes:Bacteroidetes (F:B) was plotted as a side-by-side bar graph to illustrate the variations in F:B ratio for each sample processing protocol and sampling time. \n ","description":"","filename":"6.png","url":"https://assets-eu.researchsquare.com/files/rs-129091/v1/997cae56bc93aba4077f36da.png"},{"id":4779783,"identity":"83c8e686-c83e-4241-90f5-4a0a4a0f0bb0","added_by":"auto","created_at":"2021-01-07 15:11:34","extension":"png","order_by":7,"title":"Figure 7","display":"","copyAsset":false,"role":"figure","size":50379,"visible":true,"origin":"","legend":"PCoA analysis of for aliquots analyzed from three different sample processing \nprotocols. The PCoA plot shows distinct clustering of the stool samples based on the sample processing protocol used. The legend indicates the 3 different sampling processing protocols used in this study. The plot illustrates the coordinates for each individual sample. ","description":"","filename":"7.png","url":"https://assets-eu.researchsquare.com/files/rs-129091/v1/67bc1e408f3bbeb719d986ec.png"},{"id":4338964,"identity":"3a283e8f-7f0f-453a-a89c-1c201303b29f","added_by":"auto","created_at":"2020-12-17 16:42:39","extension":"png","order_by":7,"title":"Figure 7","display":"","copyAsset":false,"role":"figure","size":50379,"visible":true,"origin":"","legend":"PCoA analysis of for aliquots analyzed from three different sample processing \nprotocols. The PCoA plot shows distinct clustering of the stool samples based on the sample processing protocol used. The legend indicates the 3 different sampling processing protocols used in this study. The plot illustrates the coordinates for each individual sample. ","description":"","filename":"7.png","url":"https://assets-eu.researchsquare.com/files/rs-129091/v1/eb48399d50385f37fa4d7eba.png"},{"id":13635011,"identity":"c43433d7-4f98-47ba-aea3-365b2c8d7f6e","added_by":"auto","created_at":"2021-09-17 08:34:32","extension":"pdf","order_by":0,"title":"","display":"","copyAsset":false,"role":"manuscript-pdf","size":1148095,"visible":true,"origin":"","legend":"","description":"","filename":"manuscript.pdf","url":"https://assets-eu.researchsquare.com/files/rs-129091/v1/3f31f21b-118f-4e20-b25b-e30bd4b37d34.pdf"},{"id":4338952,"identity":"19525452-2557-400a-b22b-313cabcf5bda","added_by":"auto","created_at":"2020-12-17 16:42:39","extension":"xlsx","order_by":1,"title":"","display":"","copyAsset":false,"role":"supplement","size":12104,"visible":true,"origin":"","legend":"","description":"","filename":"TableS1MetadataTableofDada2results.xlsx","url":"https://assets-eu.researchsquare.com/files/rs-129091/v1/7223f2e26ae94326c47850d4.xlsx"},{"id":4338958,"identity":"ca42438f-9dad-4fb4-88e0-43aaea4bd7ac","added_by":"auto","created_at":"2020-12-17 16:42:39","extension":"pdf","order_by":2,"title":"","display":"","copyAsset":false,"role":"supplement","size":8773,"visible":true,"origin":"","legend":"","description":"","filename":"FigureS1QCplots.pdf","url":"https://assets-eu.researchsquare.com/files/rs-129091/v1/2dbb6ddebece014464b64670.pdf"}],"financialInterests":"","formattedTitle":"Accelerating Gut Microbiome Research with Robust Sample Collection","fulltext":[{"header":"Full Text","content":"Due to technical limitations, full-text HTML conversion of this manuscript could not be completed. However, the latest manuscript can be downloaded and \u003ca href='/article/rs-129091/latest.pdf' target='_blank'\u003e accessed as a PDF.\u003c/a\u003e"},{"header":"Tables","content":"\u003cp\u003e\u0026nbsp;\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003e\u003cem\u003eTable 1. Wilkinson Rank Sum tests of alpha diversity metrics for the various sample processing protocols\u003c/em\u003e\u003c/strong\u003e\u003c/p\u003e\n\u003ctable border=\"1\" width=\"0\"\u003e\n\u003ctbody\u003e\n\u003ctr\u003e\n\u003ctd colspan=\"5\" width=\"863\"\u003e\n\u003cp\u003e\u003cstrong\u003eWilcoxon Rank Sum Test\u0026nbsp;\u003c/strong\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd width=\"288\"\u003e\u0026nbsp;\u003c/td\u003e\n\u003ctd colspan=\"2\" width=\"288\"\u003e\n\u003cp\u003e\u003cstrong\u003eShannon\u003c/strong\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd colspan=\"2\" width=\"288\"\u003e\n\u003cp\u003e\u003cstrong\u003eSimpson\u003c/strong\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd width=\"288\"\u003e\n\u003cp\u003e\u003cstrong\u003eCohorts\u003c/strong\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"144\"\u003e\n\u003cp\u003e\u003cstrong\u003eStatistic\u003c/strong\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"144\"\u003e\n\u003cp\u003e\u003cstrong\u003eP-value*\u003c/strong\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"144\"\u003e\n\u003cp\u003e\u003cstrong\u003eStatistic\u003c/strong\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"144\"\u003e\n\u003cp\u003e\u003cstrong\u003eP-value*\u003c/strong\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd width=\"288\"\u003e\n\u003cp\u003e\u003cstrong\u003eFresh \u0026harr; 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The gut microbiome plays a critical role in human health and disease, yet little attention has been given to optimizing collection and processing methods of human stool. \u003c/p\u003e\u003cp\u003e\u003cstrong\u003eMethods.\u003c/strong\u003e We collected the entire bowel movement from 2 healthy volunteers: one to examine stool sample heterogeneity and one to test stool sample handling parameters. Sequencing and bioinformatic analyses were used to examine the microbiome composition. \u003c/p\u003e\u003cp\u003e\u003cstrong\u003eResults.\u003c/strong\u003e The microbiome profile varied depending on where the subsample was obtained from the stool. The exterior cortex of the stool was rich in specific phyla and deficient in others while the interior core of the stool revealed opposite microbiome profiles. Sample processing also resulted in varying microbiome profiles. Homogenization and stabilization at 4°C gave superior microbial diversity profiles compared to the fresh or frozen subsamples of the same stool sample. Bacterial proliferation continued in the fresh subsample when processed at ambient temperature. Bacteroidetes proliferated and Firmicutes diminished during the 30-minute processing of fresh sample. The frozen sample had good overall diversity but Proteobacteria diminished likely because of the freeze/thaw. \u003c/p\u003e\u003cp\u003e\u003cstrong\u003eConclusions.\u003c/strong\u003e The microbiome profile is specific to the section of the stool being sampled. Stool sample collection, homogenization, and stabilization at 4°C for 24 hours provides a “neat”, high-quality sample of sufficient quantity that can be banked into aliquots with nearly identical microbial diversity profiles. This collection pipeline is essential to accelerate our understanding of the gut microbiome in health and disease\u003c/p\u003e","manuscriptTitle":"Accelerating Gut Microbiome Research with Robust Sample Collection","msid":"","msnumber":"","nonDraftVersions":[{"code":1,"date":"2020-12-17 16:42:37","doi":"10.21203/rs.3.rs-129091/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":"09b578b2-ba1c-4656-a745-c8c256da970a","owner":[],"postedDate":"December 17th, 2020","published":true,"recentEditorialEvents":[],"rejectedJournal":[],"revision":"","amendment":"","status":"posted","subjectAreas":[{"id":1515379,"name":"General Microbiology"}],"tags":[],"updatedAt":"2021-01-07T15:05:32+00:00","versionOfRecord":[],"versionCreatedAt":"2020-12-17 16:42:37","video":"","vorDoi":"","vorDoiUrl":"","workflowStages":[]},"version":"v1","identity":"rs-129091","journalConfig":"researchsquare"},"__N_SSP":true},"page":"/article/[identity]/[[...version]]","query":{"redirect":"/article/rs-129091","identity":"rs-129091","version":["v1"]},"buildId":"_2-kVJe1T_tPrBINL-cwx","isFallback":false,"isExperimentalCompile":false,"dynamicIds":[84888],"gssp":true,"scriptLoader":[]}