Isolation and characterization of diesel-degrading bacteria from hydrocarbon-contaminated sites, flower farms and soda lakes

Isolation and characterization of diesel-degrading bacteria from hydrocarbon-contaminated sites, flower farms and soda lakes | 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 Isolation and characterization of diesel-degrading bacteria from hydrocarbon-contaminated sites, flower farms and soda lakes Gessesse Kebede Bekele, Eshetu Mekonne Bogale, Tekle Tafesse Fida, and 3 more This is a preprint; it has not been peer reviewed by a journal. https://doi.org/ 10.21203/rs.3.rs-119479/v1 This work is licensed under a CC BY 4.0 License Status: Published Journal Publication published 21 Jan, 2022 Read the published version in International Journal of Microbiology → Version 1 posted You are reading this latest preprint version Abstract Background: Hydrocarbon-derived pollutants are becoming one of the most concerning ecological issues. Thus, there is a need to investigate and develop innovative, low-cost, eco-friendly, and fast techniques to reduce and/or eliminate pollutants using biological agents. The current study is conducted to isolate, characterize, and identify potential diesel-degrading bacteria. Results: Samples were collected from flower farms, lakeshores, old aged garages, asphalt, and bitumen soils and spread on selective medium (Bushnell Hass Mineral Salts Agar) containing diesel as the growth substrate. The isolates were characterized based on their growth patterns using OD measurement, biochemical testing and gravimetric analysis and identified using the Biolog database, and 16S rRNA gene sequencing techniques. Subsequently, six diesel degraders were identified and belong to Pseudomonas , Providencia , Roseomonas , Stenotrophomonas , Achromobacter , and Bacillus . Among these, based on gravimetric analysis, the three potent isolates AAUW23, AAUG11 and AAUG36 achieved 84%, 83.4%, and 83% diesel degradation efficiency, respectively, in 15 days. Consequently, the partial 16S rRNA gene sequences revealed that the two most potent bacterial strains (AAUW23 and AAUG11) were Pseudomonas aeruginosa , while AAUG36 was Bacillus subtilis . Conclusion: This study demonstrated that bacterial species isolated from hydrocarbon-contaminated and/or uncontaminated environments could be optimized to be used as potential bioremediation agents for diesel removal. Applied & Industrial Microbiology General Microbiology Biodegradation BioLog Gravimetric analysis Hydrocarbon-degradation 16S rRNA gene Figures Figure 1 Figure 2 Figure 3 Figure 4 Full Text Cite Share Download PDF Status: Published Journal Publication published 21 Jan, 2022 Read the published version in International Journal of Microbiology → 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-119479","acceptedTermsAndConditions":true,"allowDirectSubmit":true,"archivedVersions":[],"articleType":"Research","associatedPublications":[],"authors":[{"id":5626069,"identity":"f362bb29-de21-4604-8b15-de76fb9b840c","order_by":0,"name":"Gessesse Kebede Bekele","email":"","orcid":"","institution":"Addis Ababa Science and Technology University","correspondingAuthor":false,"submittingAuthor":false,"prefix":"","firstName":"Gessesse","middleName":"Kebede","lastName":"Bekele","suffix":""},{"id":5626070,"identity":"44fd1013-2dab-4576-b71d-bdc154e3f752","order_by":1,"name":"Eshetu Mekonne Bogale","email":"","orcid":"","institution":"Dire Dawa 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Gemeda","email":"data:image/png;base64,iVBORw0KGgoAAAANSUhEUgAAAZAAAAAyAQMAAABI0h/eAAAABlBMVEX///8AAABVwtN+AAAACXBIWXMAAA7EAAAOxAGVKw4bAAABI0lEQVRIie2PsWrDMBCGJQTy4uJVJbR+gkJMwBSStq9iIUiWPECGUNRFWQJePbWv0GwdHQzKIurVkCExGbrGdEmWUDklFIrqrB30wR3ouI//BIDF8k/ZRnVHad17pyFqMmCiFQLwUeyftlGTBhPwo2TnlZvJpOTlG3j0BMqq3Sj3vTiXBIy6lDtxalJCpQJOFSBE4n7LVcsgKRgmQA0odzNjSlgMnTUV+rDVNERQLCEvECZQZJQTZlZWHyWvFV96n9X+8P7wkmdaOWjF3/yRAoKj0pYuIBc8pa+pPgzyOsX8/VANa4VcziQOW65kbFawzm0kBx3hsrZRWSzKp73oedcSbard+P7uOZ+XxXbcvYqd+doY8w359Y504YZ9i8VisTTzBUB/YsvCbK8zAAAAAElFTkSuQmCC","orcid":"https://orcid.org/0000-0002-1496-2863","institution":"Addis Ababa Science and Technology University","correspondingAuthor":true,"submittingAuthor":false,"prefix":"","firstName":"Mesfin","middleName":"Tafesse","lastName":"Gemeda","suffix":""},{"id":5626074,"identity":"c2e8b5c0-f8ab-4d30-91bd-68a87476ef27","order_by":5,"name":"Fassil Assefa Tuji","email":"","orcid":"","institution":"Addis Ababa 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degradation","description":"","filename":"3.jpg","url":"https://assets-eu.researchsquare.com/files/rs-119479/v1/bfd9972210f245a7ade45f25.jpg"},{"id":3965891,"identity":"88ada318-022c-479d-a320-9dfab76d1c3b","added_by":"auto","created_at":"2020-12-02 20:00:40","extension":"jpg","order_by":4,"title":"Figure 4","display":"","copyAsset":false,"role":"figure","size":104124,"visible":true,"origin":"","legend":"Phylogenetic tree based on partial bacterial sequences of the 16S rRNA region for the two Pseudomonas isolates and one Bacillus subtilis (bold and coded with the initials ―AAU‖), and accession numbers of the 16S rRNA are followed by species names. Numbers at nodes indicated bootstrap values for each node out of 1000 bootstrap resembling. The phylogenetic tree was constructed in MEGA X using the maximum likelihood method [23] and Kimura-2 parameter model [21]. The E. coli plasmid partial sequence was used as an out-group","description":"","filename":"43.jpg","url":"https://assets-eu.researchsquare.com/files/rs-119479/v1/41e5df4470420fce0b4421c3.jpg"},{"id":17546169,"identity":"ac3e1514-5e5e-4317-9b26-fc27b37ea462","added_by":"auto","created_at":"2022-01-21 21:50:58","extension":"pdf","order_by":1,"title":"","display":"","copyAsset":false,"role":"manuscript-pdf","size":916652,"visible":true,"origin":"","legend":"","description":"","filename":"Microbiomearticleprepared02.pdf","url":"https://assets-eu.researchsquare.com/files/rs-119479/v1_covered.pdf"},{"id":13561594,"identity":"10b61acf-e330-4fcc-b9a0-0a86a198b591","added_by":"auto","created_at":"2021-09-17 03:09:40","extension":"pdf","order_by":1,"title":"","display":"","copyAsset":false,"role":"manuscript-pdf","size":911541,"visible":true,"origin":"","legend":"","description":"","filename":"Microbiomearticleprepared02.pdf","url":"https://assets-eu.researchsquare.com/files/rs-119479/v1_covered.pdf"},{"id":3965892,"identity":"872d43ea-42de-481c-b027-a9879d5619c3","added_by":"auto","created_at":"2020-12-02 20:00:52","extension":"pdf","order_by":1,"title":"","display":"","copyAsset":false,"role":"manuscript-pdf","size":948640,"visible":true,"origin":"","legend":"","description":"","filename":"Microbiomearticleprepared02.pdf","url":"https://assets-eu.researchsquare.com/files/rs-119479/v1_stamped.pdf"}],"financialInterests":"","formattedTitle":"Isolation and characterization of diesel-degrading bacteria from hydrocarbon-contaminated sites, flower farms and soda lakes","fulltext":[{"header":"Full Text","content":"\u003cp\u003eThis preprint is available for \u003ca href='/article/rs-119479/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":false,"highlight":"","institution":"","isAcceptedByJournal":true,"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":"Biodegradation, BioLog, Gravimetric analysis, Hydrocarbon-degradation, 16S rRNA gene","lastPublishedDoi":"10.21203/rs.3.rs-119479/v1","lastPublishedDoiUrl":"https://doi.org/10.21203/rs.3.rs-119479/v1","license":{"name":"CC BY 4.0","url":"https://creativecommons.org/licenses/by/4.0/"},"manuscriptAbstract":"\u003cp\u003e\u003cstrong\u003eBackground: \u003c/strong\u003eHydrocarbon-derived pollutants are becoming one of the most concerning ecological issues. Thus, there is a need to investigate and develop innovative, low-cost, eco-friendly, and fast techniques to reduce and/or eliminate pollutants using biological agents. The current study is conducted to isolate, characterize, and identify potential diesel-degrading bacteria.\u003c/p\u003e\u003cp\u003e\u003cstrong\u003eResults: \u003c/strong\u003eSamples were collected from flower farms, lakeshores, old aged garages, asphalt, and bitumen soils and spread on selective medium (Bushnell Hass Mineral Salts Agar) containing diesel as the growth substrate. The isolates were characterized based on their growth patterns using OD measurement, biochemical testing and gravimetric analysis and identified using the Biolog database, and 16S rRNA gene sequencing techniques. Subsequently, six diesel degraders were identified and belong to Pseudomonas , Providencia , Roseomonas , Stenotrophomonas , Achromobacter , and Bacillus . Among these, based on gravimetric analysis, the three potent isolates AAUW23, AAUG11 and AAUG36 achieved 84%, 83.4%, and 83% diesel degradation efficiency, respectively, in 15 days. Consequently, the partial 16S rRNA gene sequences revealed that the two most potent bacterial strains (AAUW23 and AAUG11) were Pseudomonas aeruginosa , while AAUG36 was Bacillus subtilis . \u003c/p\u003e\u003cp\u003e\u003cstrong\u003eConclusion:\u003c/strong\u003e This study demonstrated that bacterial species isolated from hydrocarbon-contaminated and/or uncontaminated environments could be optimized to be used as potential bioremediation agents for diesel removal.\u003c/p\u003e","manuscriptTitle":"Isolation and characterization of diesel-degrading bacteria from hydrocarbon-contaminated sites, flower farms and soda lakes","msid":"","msnumber":"","nonDraftVersions":[{"code":1,"date":"2020-12-02 20:00:10","doi":"10.21203/rs.3.rs-119479/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":"13eaeaa7-1d02-40a0-85fc-16c24c31553f","owner":[],"postedDate":"December 2nd, 2020","published":true,"recentEditorialEvents":[],"rejectedJournal":[],"revision":"","amendment":"","status":"published-in-journal","subjectAreas":[{"id":1303877,"name":"Applied \u0026 Industrial Microbiology"},{"id":1303878,"name":"General Microbiology"}],"tags":[],"updatedAt":"2022-01-21T21:50:46+00:00","versionOfRecord":{"articleIdentity":"rs-119479","link":"https://doi.org/10.1155/2022/5655767","journal":{"identity":"international-journal-of-microbiology","isVorOnly":true,"title":"International Journal of Microbiology"},"publishedOn":"2022-01-21 21:50:46","publishedOnDateReadable":"January 21st, 2022"},"versionCreatedAt":"2020-12-02 20:00:10","video":"","vorDoi":"10.1155/2022/5655767","vorDoiUrl":"https://doi.org/10.1155/2022/5655767","workflowStages":[]},"version":"v1","identity":"rs-119479","journalConfig":"researchsquare"},"__N_SSP":true},"page":"/article/[identity]/[[...version]]","query":{"redirect":"/article/rs-119479","identity":"rs-119479","version":["v1"]},"buildId":"cBFmMYwuxLRRLfASyISRj","isFallback":false,"isExperimentalCompile":false,"dynamicIds":[84888],"gssp":true,"scriptLoader":[]}