Nitric Oxide-Targeted Anammox (NOTA): A Novel Pathway for Nitrogen Flux Regulation

Nitric Oxide-Targeted Anammox (NOTA): A Novel Pathway for Nitrogen Flux Regulation | 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 Article Nitric Oxide-Targeted Anammox (NOTA): A Novel Pathway for Nitrogen Flux Regulation Sankar Ganesh Palani, Hemapriya Srinivasan, Anand N This is a preprint; it has not been peer reviewed by a journal. https://doi.org/ 10.21203/rs.3.rs-8962830/v1 This work is licensed under a CC BY 4.0 License Status: Under Review Version 1 posted You are reading this latest preprint version Abstract Closed landfill leachate (CLL) contains elevated levels of ammoniacal nitrogen and humic substances that have accumulated over decades, disrupting global nitrogen and carbon biogeochemical cycles. This study examines the nitric oxide-targeted anammox (NOTA), a novel biochemical pathway for nitrogen and carbon recovery from CLL, utilizing waste-activated sludge (WAS) as the inoculum. Notably, during the anammox process the nitric oxide pathway was preferred over the hydrazine pathway. Aerated (ANR) and stirred NOTA reactors (SNR) exhibited ammoniacal nitrogen removal efficiencies of 98% and 92%, respectively, and biological oxygen demand (BOD) removal rates of 60.2% and 43.4%, demonstrating the impact of active (ANR) and passive aeration (SNR) on NOTA performance. Metagenomic analysis revealed microbial community shifts, with an increased presence of nitrogen and carbon degrading bacteria. These findings contribute to the advancement of sustainable CLL treatment with minimal aeration and reinforce the role of NOTA in enhancing biological nitrogen and carbon cycling. Biological sciences/Biotechnology/Environmental biotechnology Earth and environmental sciences/Biogeochemistry/Element cycles Full Text Additional Declarations There is NO Competing Interest. Cite Share Download PDF Status: Under Review 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-8962830","acceptedTermsAndConditions":true,"allowDirectSubmit":false,"archivedVersions":[],"articleType":"Article","associatedPublications":[],"authors":[{"id":607771682,"identity":"4917a88a-974c-41e9-b632-e632e993e395","order_by":0,"name":"Sankar Ganesh Palani","email":"data:image/png;base64,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","orcid":"https://orcid.org/0000-0002-9278-8514","institution":"Birla Institute of Technology and Science, Pilani, Hyderabad Campus, India","correspondingAuthor":true,"prefix":"","firstName":"Sankar","middleName":"Ganesh","lastName":"Palani","suffix":""},{"id":607771683,"identity":"3d2e4dfc-cf9c-4535-9e9c-476bb9f33df8","order_by":1,"name":"Hemapriya Srinivasan","email":"","orcid":"","institution":"Birla Institute of Technology and Science, Pilani, Hyderabad Campus, India","correspondingAuthor":false,"prefix":"","firstName":"Hemapriya","middleName":"","lastName":"Srinivasan","suffix":""},{"id":607771684,"identity":"9d609868-35e1-4682-bb92-282019762ea1","order_by":2,"name":"Anand N","email":"","orcid":"","institution":"Sardar Patel Renewable Energy Research Institute (SPRERI)","correspondingAuthor":false,"prefix":"","firstName":"Anand","middleName":"","lastName":"N","suffix":""}],"badges":[],"createdAt":"2026-02-25 04:00:20","currentVersionCode":1,"declarations":"","doi":"10.21203/rs.3.rs-8962830/v1","doiUrl":"https://doi.org/10.21203/rs.3.rs-8962830/v1","draftVersion":[],"editorialEvents":[],"editorialNote":"","failedWorkflow":false,"files":[{"id":106093073,"identity":"e8460f2f-cf77-486f-96f2-defcc464222f","added_by":"auto","created_at":"2026-04-03 11:33:33","extension":"pdf","order_by":1,"title":"","display":"","copyAsset":false,"role":"manuscript-pdf","size":1881954,"visible":true,"origin":"","legend":"","description":"","filename":"Manuscript.pdf","url":"https://assets-eu.researchsquare.com/files/rs-8962830/v1_covered_84eb0dde-a25b-4b64-b720-2baa77aa6464.pdf"}],"financialInterests":"There is \u003cb\u003eNO\u003c/b\u003e Competing Interest.","formattedTitle":"Nitric Oxide-Targeted Anammox (NOTA): A Novel Pathway for Nitrogen Flux Regulation","fulltext":[],"fulltextSource":"","fullText":"","funders":[],"hasAdminPriorityOnWorkflow":false,"hasManuscriptDocX":false,"hasOptedInToPreprint":true,"hasPassedJournalQc":"","hasAnyPriority":false,"hideJournal":false,"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":"commssustain","isNatureJournal":false,"hasQc":false,"allowDirectSubmit":false,"externalIdentity":"commssustain","sideBox":"Learn more about [Communications Sustainability](https://www.nature.com/commssustain/)","snPcode":"44458","submissionUrl":"https://mts-commssustain.nature.com","title":"Communications Sustainability","twitterHandle":"","acdcEnabled":true,"dfaEnabled":true,"editorialSystem":"ejp","reportingPortfolio":"Nature","inReviewEnabled":true,"inReviewRevisionsEnabled":true},"keywords":"","lastPublishedDoi":"10.21203/rs.3.rs-8962830/v1","lastPublishedDoiUrl":"https://doi.org/10.21203/rs.3.rs-8962830/v1","license":{"name":"CC BY 4.0","url":"https://creativecommons.org/licenses/by/4.0/"},"manuscriptAbstract":"Closed landfill leachate (CLL) contains elevated levels of ammoniacal nitrogen and humic substances that have accumulated over decades, disrupting global nitrogen and carbon biogeochemical cycles. 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