Microbiome and Metagenome Analyses of a Closed Habitat During Human Occupation

Microbiome and Metagenome Analyses of a Closed Habitat During Human Occupation | 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 Microbiome Announcement Microbiome and Metagenome Analyses of a Closed Habitat During Human Occupation Ganesh Babu Malli Mohan, Ceth Parker, Camilla Urbaniak, Nitin Singh, and 5 more This is a preprint; it has not been peer reviewed by a journal. https://doi.org/ 10.21203/rs.3.rs-21521/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: Microbial contamination during long-term confinements of space exploration present potential risks for both crew members and spacecraft life support systems. As NASA prepares for manned missions beyond low Earth orbit, deeper into the solar system, the monitoring of microbial populations within closed human habitation will be necessary to ensure the safety of both the crew and the spacecraft. NASA’s Johnson Space Center recently developed a microbial swab kit, designed specifically to be used during astronaut Extravehicular Activity (EVA). The EVA swab kit was designed in such a way that it could be held easily within an astronaut’s bulky glove and or by a robot’s manipulator, making it suitable for microbial sample collection in remote and extreme environments. The previously tested (in laboratory and controlled settings) EVA swab kit was used in this study to sample various surfaces from a submerged, closed, analog habitat in order to characterize the microbial populations in this unique human habitat. Results: Samples were collected from various locations across the habitat of which were constructed from various surface materials (linoleum, dry wall, particle board, glass, and metal) and microbial populations examined by culture, qPCR, microbiome 16S rRNA gene sequencing and shot gun metagenomics. Propidium monoazide treated samples identified the viable/intact microbial population of the habitat. The cultivable microbial population ranged from below the detection limit (BDL) to 10 6 CFU/sample and their identity was characterized using Sanger sequencing. Next-generation sequencing (NGS; both 16S rRNA amplicon and shotgun) were used to characterize the microbial dynamics, community profiles and functional attributes (metabolism, virulence, and antimicrobial resistance). The 16S rRNA amplicon sequencing revealed abundance of viable Actinobacteria ( Brevibacterium , Nesternkonia, Mycobacterium, Pseudonocardia and Corynebacterium ), Firmicutes ( Virgibacillus , Staphylococcus and Oceanobacillus ) and Proteobacteria (esp. Acinetobacter ) on linoleum, dry wall, and particle board (LDP) surfaces, while members of Firmicutes ( Leuconostocaceae ) and Proteobacteria ( Enterobacteriaceae ) were high on the glass/metal surfaces. Non-metric multidimensional scaling (NMDS) determined from both 16S rRNA and metagenomic analyses revealed differential microbial speciation between LDP surfaces and glass/metal surfaces. The shotgun metagenomics sequencing showed bacterial predominance of Brevibacterium (53.6%), Brachybacterium (7.8%), Pseudonocardia (9.9%), Mycobacterium (3.7%), and Staphylococcus (2.1%); while fungal analyses revealed Aspergillus and Penicillium dominance. Conclusion: This study provides the first assessment of monitoring cultivable and viable microorganisms on surfaces within a submerged, closed, analog habitat. The analyses presented herein suggests that the surface material plays a role in microbial community structure as the microbial populations differed between LDP and metal/glass surfaces. The metal/glass surfaces had less complex community, lower bio-burden, and more closely resembled the controls. These results indicated that material choice is crucial when building closed habitats, even if they are simply analogs. Finally, while a few species were associated with previously cultivated isolates from the International Space Station and MIR spacecraft, the majority of the microbial ecology of the submerged Analog habitat differs greatly from that of previously studied analog habitats. General Microbiology xtravehicular Activity Analog habitat Microbiome Microbial diversity Functional Metagenomics Spacecraft microbiome Closed habitat Figures Figure 1 Figure 2 Figure 3 Figure 4 Figure 5 Figure 6 Figure 7 Full Text Supplementary Files AnaloghabitatSupplFiguresS1toS4.pdf AnalogDatasetS116Samplicon.xlsx AnalogDatasetS2metagenome.xlsx 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-21521","acceptedTermsAndConditions":true,"allowDirectSubmit":true,"archivedVersions":[],"articleType":"Microbiome Announcement","associatedPublications":[],"authors":[{"id":471036,"identity":"be1838fe-f1fd-4871-9c46-149d677ec449","order_by":1,"name":"Ganesh Babu Malli Mohan","email":"","orcid":"","institution":"NASA Jet Propulsion Laboratory","correspondingAuthor":false,"submittingAuthor":false,"prefix":"","firstName":"Ganesh","middleName":"Babu Malli","lastName":"Mohan","suffix":""},{"id":471037,"identity":"4d558930-0444-4dba-adf7-5732da753465","order_by":2,"name":"Ceth Parker","email":"","orcid":"","institution":"NASA Jet Propulsion Laboratory","correspondingAuthor":false,"submittingAuthor":false,"prefix":"","firstName":"Ceth","middleName":"","lastName":"Parker","suffix":""},{"id":471038,"identity":"6acc1439-6231-4b60-84f1-d58c0d9e0474","order_by":3,"name":"Camilla Urbaniak","email":"","orcid":"","institution":"NASA Jet Propulsion Laboratory","correspondingAuthor":false,"submittingAuthor":false,"prefix":"","firstName":"Camilla","middleName":"","lastName":"Urbaniak","suffix":""},{"id":471039,"identity":"e833cbe9-2f5a-4c8c-8a68-3f84087d313b","order_by":4,"name":"Nitin Singh","email":"","orcid":"","institution":"NASA Jet Propulsion Laboratory","correspondingAuthor":false,"submittingAuthor":false,"prefix":"","firstName":"Nitin","middleName":"","lastName":"Singh","suffix":""},{"id":471040,"identity":"4d9ebabc-b582-4fc8-8dd2-1693cef7ed34","order_by":5,"name":"Anthony Hood","email":"","orcid":"","institution":"NASA Johnson Space Center","correspondingAuthor":false,"submittingAuthor":false,"prefix":"","firstName":"Anthony","middleName":"","lastName":"Hood","suffix":""},{"id":471041,"identity":"d7711514-4c50-4cc7-bcea-d5fc4a6d42b3","order_by":6,"name":"Jeremiah Minich","email":"","orcid":"","institution":"University of California San Diego","correspondingAuthor":false,"submittingAuthor":false,"prefix":"","firstName":"Jeremiah","middleName":"","lastName":"Minich","suffix":""},{"id":471042,"identity":"89125062-3f1a-447a-aa14-858a345d8bee","order_by":7,"name":"Rob Knight","email":"","orcid":"","institution":"University of California San Diego","correspondingAuthor":false,"submittingAuthor":false,"prefix":"","firstName":"Rob","middleName":"","lastName":"Knight","suffix":""},{"id":471043,"identity":"d99cc28a-157e-4870-a08d-85e0917c50af","order_by":8,"name":"Michelle A Rucker","email":"","orcid":"","institution":"NASA Johnson Space Center","correspondingAuthor":false,"submittingAuthor":false,"prefix":"","firstName":"Michelle","middleName":"A","lastName":"Rucker","suffix":""},{"id":471044,"identity":"ef9179d0-9a56-463b-9e8c-3b3fa9aad6ec","order_by":9,"name":"Kasthuri Venkateswaran","email":"data:image/png;base64,iVBORw0KGgoAAAANSUhEUgAAAZAAAAAyAQMAAABI0h/eAAAABlBMVEX///8AAABVwtN+AAAACXBIWXMAAA7EAAAOxAGVKw4bAAAA80lEQVRIie3SMWrDMBSA4Sc0eHngVcaQXEFF0DG+ik0gXQsdA42yuEsOoGOkW8dnBPaiBLJ5dMgFXDp2qeviVU23QvSDhBD6hgcCCIX+YQJo2IcVAzH6ExGJJriWwA+RBFeSZOvuLo9u8ayqAxF7W2Sxjk4X9JAUnFKmXYp7OubE3LIwhE/KR2ZQr1Ls+UCcJFbyHAhXifGTh0/sN0LpkWyy+W8kBVtzbK2QMBLL9hTVoveNr61N0TWJ+Z6lKJvi1SKXPiLaavuB9TqOzaHq3st1Nmtezl3uISBoOjiA8SVH6QPDR9HTYTddRZ2fhEKh0K31BThoV6oktmClAAAAAElFTkSuQmCC","orcid":"https://orcid.org/0000-0002-6742-0873","institution":"NASA Jet Propulsion Laboratory","correspondingAuthor":true,"submittingAuthor":false,"prefix":"","firstName":"Kasthuri","middleName":"","lastName":"Venkateswaran","suffix":""}],"badges":[],"createdAt":"2020-04-06 10:42:10","currentVersionCode":1,"declarations":"","doi":"10.21203/rs.3.rs-21521/v1","doiUrl":"https://doi.org/10.21203/rs.3.rs-21521/v1","draftVersion":[],"editorialEvents":[],"editorialNote":"","failedWorkflow":false,"files":[{"id":867481,"identity":"96d45f93-3d1d-4832-9cad-bfee4fe0249d","added_by":"auto","created_at":"2020-04-08 14:46:50","extension":"png","order_by":1,"title":"Figure 1","display":"","copyAsset":false,"role":"figure","size":1690619,"visible":true,"origin":"","legend":"Location and sampling tool kit feature for surface sampling\n (A) Cantilever swab tool kit storage canister box and b) Swab head attached to the cantilever tool kit. (B) 2D outline of the Analog habitat and sampling location denoted in numbers. (C)\nPhotography of Analog habitat sampling locations and the red circle represents sample\ncollecting area.","description":"","filename":"1.PNG","url":"https://assets-eu.researchsquare.com/files/rs-21521/v1/1.PNG"},{"id":867484,"identity":"2f74e9c4-e4a5-4ac3-8af5-ca67ae114e6f","added_by":"auto","created_at":"2020-04-08 14:46:51","extension":"png","order_by":2,"title":"Figure 2","display":"","copyAsset":false,"role":"figure","size":170441,"visible":true,"origin":"","legend":"Culture dependent and independent analysis from Analog habitat surface sample\n(A) Abundance of cultivable bacteria and fungi. Each dot in a column represents Analog habitat\n location sampled. No statistically significant differences in abundances were observed amongst\n flight missions and between bacteria and fungi (one way- ANOVA, p\u003e0.01; ANOVA).\n (B) The relative light unit of ATP counts for total (round dot) and intracellular ATP (square dot).\nNo statistically significant differences in abundances were observed amongst flight missions and\nbetween bacteria and fungi (one way- ANOVA, p\u003e0.01; ANOVA).\n(C) The qPCR based microbial burden (total; non-PMA and viable; PMA) of various Analog\nhabitat surface sample. The gene copies were measured by targeting 16S rRNA gene (bacteria)\nand ITS gene (fungi).","description":"","filename":"2.PNG","url":"https://assets-eu.researchsquare.com/files/rs-21521/v1/2.PNG"},{"id":867486,"identity":"9aeb1cda-0065-4fc1-b6de-3252b6412368","added_by":"auto","created_at":"2020-04-08 14:46:51","extension":"png","order_by":3,"title":"Figure 3","display":"","copyAsset":false,"role":"figure","size":125916,"visible":true,"origin":"","legend":"Multi-dimensional scaling plots of 16S rRNA gene amplicon sequencing data\nNMDS ordination showing 99% confidence interval ellipses of non-PMA treated (left panels)\n and PMA treated (right panels) grouped based on surface material (A) and site categories (B).\nThe various samples collected from across the Analog habitat are indicated by a dot and the closer the dots are to each other the more similar their bacterial composition is in terms of types and number of bacteria.","description":"","filename":"3.PNG","url":"https://assets-eu.researchsquare.com/files/rs-21521/v1/3.PNG"},{"id":867487,"identity":"697f6391-fba4-42d6-bc10-abdfca7e699f","added_by":"auto","created_at":"2020-04-08 14:46:51","extension":"png","order_by":4,"title":"Figure 4","display":"","copyAsset":false,"role":"figure","size":394575,"visible":true,"origin":"","legend":"Relative abundances of bacteria detected by 16S rRNA gene amplicon sequencing\nThe relative abundances of bacterial taxa identified in various samples across the Analog\nhabitat were visualize by bar plots. Each bar represents a specific sample and each colored box\na particular taxon. The height of the colored box represents the relative abundance of that\nparticular taxon within the sample. Taxa present in less than 1% abundance in a given sample\nare displayed in the “remaining fraction” at the top of the graph (gray box). The legend is read\nfrom bottom to top, with the bottom taxon in the legend corresponding to the bottom taxon\non the graph. Non PMA treated samples are displayed in (A) and the PMA treated samples in (B).","description":"","filename":"4.PNG","url":"https://assets-eu.researchsquare.com/files/rs-21521/v1/4.PNG"},{"id":867488,"identity":"8e85ee47-1f72-4d10-8942-e431e9de7537","added_by":"auto","created_at":"2020-04-08 14:46:51","extension":"png","order_by":5,"title":"Figure 5","display":"","copyAsset":false,"role":"figure","size":532591,"visible":true,"origin":"","legend":"Differential bacterial composition among various types of surfaces\nPie chart of the relative abundances of bacteria detected in the Analog habitat. The sequences obtained were summarized to the genus level. In total, 52 taxa were detected but the 10 most\nabundant ones are just displayed in the legends. The pie graphs are separated based on-site\n categories; LDP samples (left panels) and metal/glass (right panels) and treatment group: no\nPMA (A) and PMA (B).","description":"","filename":"5.PNG","url":"https://assets-eu.researchsquare.com/files/rs-21521/v1/5.PNG"},{"id":867489,"identity":"2e606e1e-d1b8-48dc-989e-c77d1c631cca","added_by":"auto","created_at":"2020-04-08 14:46:52","extension":"png","order_by":6,"title":"Figure 6","display":"","copyAsset":false,"role":"figure","size":238656,"visible":true,"origin":"","legend":"Metagenomic sequencing analysis of bacteria of the Analog habitat\nCluster dendrogram of Euclidean distances was performed on PMA untreated (A Top) and PMA\ntreated samples (B Top). NMDS ordination showing the 95% confidence interval ellipse based\non Unifrac distances matrix all microbial species (bacteria and fungi) from both PMA untreated \n(Middle left panel) and treated (Middle right panel) samples. Similar 926 treatment was performed\nfor all bacterial species and NMDS ordination plots are depicted for PMA untreated (Bottom\n left panel) and PMA treated (Bottom right panel) samples. The samples collected from various\nAnalog habitat surfaces was indicated by various color circles.","description":"","filename":"6.PNG","url":"https://assets-eu.researchsquare.com/files/rs-21521/v1/6.PNG"},{"id":867490,"identity":"fd60baa3-fba9-4b34-861d-5c8326eca7a4","added_by":"auto","created_at":"2020-04-08 14:46:52","extension":"png","order_by":7,"title":"Figure 7","display":"","copyAsset":false,"role":"figure","size":1116554,"visible":true,"origin":"","legend":"Metagenomic sequencing analysis of bacteria of the Analog habitat\nHeat map showing the relative abundance of each antimicrobial associated gene (Top) and virulence associated genes (Bottom) detected in each sample collected.","description":"","filename":"7.PNG","url":"https://assets-eu.researchsquare.com/files/rs-21521/v1/7.PNG"},{"id":913487,"identity":"7b1867b2-abf2-48a0-bfe7-a40e962cc15a","added_by":"auto","created_at":"2020-04-16 14:54:50","extension":"pdf","order_by":0,"title":"","display":"","copyAsset":false,"role":"manuscript-pdf","size":555748,"visible":true,"origin":"","legend":"","description":"","filename":"AnalogmanuscriptApril52020.pdf","url":"https://assets-eu.researchsquare.com/files/rs-21521/v1/Analog manuscript April 5, 2020.pdf"},{"id":867491,"identity":"77178f78-1d48-4cb2-b02b-b3ed0b6eca06","added_by":"auto","created_at":"2020-04-08 14:46:54","extension":"pdf","order_by":0,"title":"","display":"","copyAsset":false,"role":"manuscript-pdf","size":534048,"visible":true,"origin":"","legend":"","description":"","filename":"AnalogmanuscriptApril52020.pdf","url":"https://assets-eu.researchsquare.com/files/rs-21521/v1/Manuscript.pdf"},{"id":867482,"identity":"39b1dbaf-e2c8-4024-8f27-2d83d03b8114","added_by":"auto","created_at":"2020-04-08 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14:46:51","extension":"pdf","order_by":0,"title":"","display":"","copyAsset":false,"role":"supplement","size":386275,"visible":true,"origin":"","legend":"","description":"","filename":"AnaloghabitatSupplFiguresS1toS4.pdf","url":"https://assets-eu.researchsquare.com/files/rs-21521/v1/Analog_habitat_Suppl Figures_S1 to S4.pdf"},{"id":867483,"identity":"4cd9d528-ba43-4e2e-9a00-607c2206ba78","added_by":"auto","created_at":"2020-04-08 14:46:50","extension":"xlsx","order_by":0,"title":"","display":"","copyAsset":false,"role":"supplement","size":18113,"visible":true,"origin":"","legend":"","description":"","filename":"AnalogDatasetS116Samplicon.xlsx","url":"https://assets-eu.researchsquare.com/files/rs-21521/v1/Analog Dataset S1 (16S amplicon).xlsx"},{"id":867480,"identity":"de803988-59eb-4ca3-8137-ebe918389f1c","added_by":"auto","created_at":"2020-04-08 14:46:50","extension":"xlsx","order_by":0,"title":"","display":"","copyAsset":false,"role":"supplement","size":131411,"visible":true,"origin":"","legend":"","description":"","filename":"AnalogDatasetS2metagenome.xlsx","url":"https://assets-eu.researchsquare.com/files/rs-21521/v1/Analog Dataset S2 (metagenome).xlsx"}],"financialInterests":"","formattedTitle":"Microbiome and Metagenome Analyses of a Closed Habitat During Human Occupation","fulltext":[{"header":"Full Text","content":"\u003cp\u003eThis preprint is available for \u003ca href='/article/rs-21521/latest.pdf' target='_blank'\u003edownload as a PDF\u003c/a\u003e.\u003c/p\u003e"}],"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":false,"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":"xtravehicular Activity, Analog habitat, Microbiome, Microbial diversity, Functional Metagenomics, Spacecraft microbiome, Closed habitat","lastPublishedDoi":"10.21203/rs.3.rs-21521/v1","lastPublishedDoiUrl":"https://doi.org/10.21203/rs.3.rs-21521/v1","license":{"name":"CC BY 4.0","url":"https://creativecommons.org/licenses/by/4.0/"},"manuscriptAbstract":"\u003cp\u003eBackground: Microbial contamination during long-term confinements of space exploration present potential risks for both crew members and spacecraft life support systems. As NASA prepares for manned missions beyond low Earth orbit, deeper into the solar system, the monitoring of microbial populations within closed human habitation will be necessary to ensure the safety of both the crew and the spacecraft. NASA’s Johnson Space Center recently developed a microbial swab kit, designed specifically to be used\u0026nbsp;during astronaut Extravehicular Activity (EVA). The EVA swab kit was designed in such a way that it could be held easily within an astronaut’s bulky glove and or by a robot’s manipulator, making it suitable for microbial sample collection in remote and extreme environments. The previously tested (in laboratory and controlled settings) EVA swab kit was used in this study to sample various surfaces from a submerged, closed, analog habitat in order to characterize the microbial populations in this unique human habitat.\u0026nbsp;\u003c/p\u003e\u003cp\u003eResults: Samples were collected from various locations across the habitat of which were constructed from various surface materials (linoleum, dry wall, particle board, glass, and metal) and microbial populations examined by culture, qPCR, microbiome 16S rRNA gene sequencing and shot gun metagenomics. Propidium monoazide treated samples identified the viable/intact microbial population of the habitat. The cultivable microbial population ranged from below the detection limit (BDL) to 10 6 CFU/sample and their identity was characterized using Sanger sequencing. Next-generation sequencing (NGS; both 16S rRNA amplicon and shotgun) were used to characterize the microbial dynamics, community profiles and functional attributes (metabolism, virulence, and antimicrobial resistance). The 16S rRNA amplicon sequencing revealed abundance of viable \u003cem\u003eActinobacteria \u003c/em\u003e( \u003cem\u003eBrevibacterium , Nesternkonia, Mycobacterium, Pseudonocardia\u003c/em\u003e and \u003cem\u003eCorynebacterium \u003c/em\u003e), \u003cem\u003eFirmicutes \u003c/em\u003e( \u003cem\u003eVirgibacillus , Staphylococcus\u003c/em\u003e and \u003cem\u003eOceanobacillus \u003c/em\u003e) and Proteobacteria (esp. \u003cem\u003eAcinetobacter \u003c/em\u003e) on linoleum, dry wall, and particle board (LDP) surfaces, while members of \u003cem\u003eFirmicutes \u003c/em\u003e( \u003cem\u003eLeuconostocaceae \u003c/em\u003e) and Proteobacteria ( \u003cem\u003eEnterobacteriaceae \u003c/em\u003e) were high on the glass/metal surfaces. Non-metric multidimensional scaling (NMDS) determined from both 16S rRNA and metagenomic analyses revealed differential microbial speciation between LDP surfaces and glass/metal surfaces. The shotgun metagenomics sequencing showed bacterial predominance of \u003cem\u003eBrevibacterium \u003c/em\u003e(53.6%), \u003cem\u003eBrachybacterium \u003c/em\u003e(7.8%), \u003cem\u003ePseudonocardia \u003c/em\u003e(9.9%), \u003cem\u003eMycobacterium \u003c/em\u003e(3.7%), and \u003cem\u003eStaphylococcus \u003c/em\u003e(2.1%); while fungal analyses revealed \u003cem\u003eAspergillus \u003c/em\u003eand \u003cem\u003ePenicillium \u003c/em\u003edominance. \u003c/p\u003e\u003cp\u003eConclusion: This study provides the first assessment of monitoring cultivable and viable microorganisms on surfaces within a submerged, closed, analog habitat. The analyses presented herein suggests that the surface material plays a role in microbial community structure as the microbial populations differed between LDP and metal/glass surfaces. The metal/glass surfaces had less complex community, lower bio-burden, and more closely resembled the controls. These results indicated that material choice is crucial when building closed habitats, even if they are simply analogs. Finally, while a few species were associated with previously cultivated isolates from the International Space Station and MIR spacecraft, the majority of the microbial ecology of the submerged Analog habitat differs greatly from that of previously studied analog habitats.\u003c/p\u003e","manuscriptTitle":"Microbiome and Metagenome Analyses of a Closed Habitat During Human Occupation","msid":"","msnumber":"","nonDraftVersions":[{"code":1,"date":"2020-04-08 14:46:47","doi":"10.21203/rs.3.rs-21521/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":"bf60d9e7-28f0-4c1d-9fe1-21b6ec944776","owner":[],"postedDate":"April 8th, 2020","published":true,"recentEditorialEvents":[],"rejectedJournal":[],"revision":"","amendment":"","status":"posted","subjectAreas":[{"id":80853,"name":"General Microbiology"}],"tags":[],"updatedAt":"2020-04-16T14:54:48+00:00","versionOfRecord":[],"versionCreatedAt":"2020-04-08 14:46:47","video":"","vorDoi":"","vorDoiUrl":"","workflowStages":[]},"version":"v1","identity":"rs-21521","journalConfig":"researchsquare"},"__N_SSP":true},"page":"/article/[identity]/[[...version]]","query":{"redirect":"/article/rs-21521","identity":"rs-21521","version":["v1"]},"buildId":"cBFmMYwuxLRRLfASyISRj","isFallback":false,"isExperimentalCompile":false,"dynamicIds":[84888],"gssp":true,"scriptLoader":[]}