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Progress has been hindered by the difficulty in cultivating anaerobic benzene-degrading enrichment cultures. Our laboratory has sustained a methanogenic enrichment culture harboring Desulfobacterota ORM2, a benzene fermenter distinct from any known genus but related to other known or predicted benzene degraders. ORM2\u2019s slow doubling time (\u223c30 days) and extended lag phase after inoculation complicate its study. We developed a fluorescent in situ hybridization (FISH) probe for ORM2, revealing rod-shaped cells of variable length that tend to cluster with other organisms, particularly methanogens. Microscopy and genomic evidence suggest that ORM2 may produce extracellular polymeric substances, facilitating cell aggregation and possibly consuming energy that contributes to the lag phase. Interestingly, higher benzene concentrations (90\u2013120 mg\/L) appeared to reduce cell aggregation. This study visualized the cells of Desulfobacterota ORM2 within a methanogenic community, offering insights into spatial organization and potential strategies to enhance its growth rate.<\/jats:p>","DOI":"10.1021\/acs.est.4c08254","type":"journal-article","created":{"date-parts":[[2025,1,3]],"date-time":"2025-01-03T12:41:31Z","timestamp":1735908091000},"page":"591-602","source":"Crossref","is-referenced-by-count":4,"title":["Visualization of\nSyntrophic Benzene-Fermenting\n                    <i>Desulfobacterota<\/i>\n                    ORM2\nin a Methanogenic Enrichment Culture\nUsing Fluorescence In Situ Hybridization"],"prefix":"10.1021","volume":"59","author":[{"ORCID":"https:\/\/orcid.org\/0009-0004-9150-6249","authenticated-orcid":true,"given":"Xu","family":"Chen","sequence":"first","affiliation":[{"name":"University of Toronto , , , ,","place":["Toronto, Ontario, Canada, M5S 3E5"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-3430-0792","authenticated-orcid":true,"given":"Courtney R. A.","family":"Toth","sequence":"additional","affiliation":[{"name":"University of Toronto , , , ,","place":["Toronto, Ontario, Canada, M5S 3E5"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Shen","family":"Guo","sequence":"additional","affiliation":[{"name":"University of Toronto , , , ,","place":["Toronto, Ontario, Canada, M5S 3E5"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Fei","family":"Luo","sequence":"additional","affiliation":[{"name":"University of Toronto , , , ,","place":["Toronto, Ontario, Canada, M5S 3E5"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-9319-3988","authenticated-orcid":true,"given":"Jane","family":"Howe","sequence":"additional","affiliation":[{"name":"University of Toronto , , 184 College Street , , ,","place":["Toronto, Ontario, Canada, M5S 3E4"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Camilla L.","family":"Nesbo\u0337","sequence":"additional","affiliation":[{"name":"University of Toronto , , , ,","place":["Toronto, Ontario, Canada, M5S 3E5"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-8071-338X","authenticated-orcid":true,"given":"Elizabeth A.","family":"Edwards","sequence":"additional","affiliation":[{"name":"University of Toronto , , , ,","place":["Toronto, Ontario, Canada, M5S 3E5"]}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"316","published-online":{"date-parts":[[2025,1,3]]},"reference":[{"issue":"9","key":"2026081212021241300_cit1","doi-asserted-by":"publisher","first-page":"2923","DOI":"10.1111\/1462-2920.13121","article-title":"Cultivating Microbial Dark Matter\nin Benzene-Degrading Methanogenic Consortia","volume":"18","author":"Luo","year":"2016","journal-title":"Env. 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Biotechnol."},{"issue":"1\u20134","key":"2026081212021241300_cit5","doi-asserted-by":"publisher","first-page":"165","DOI":"10.1007\/s11270-008-9643-x","article-title":"Benzene Biodegradation under Anaerobic Conditions Coupled\nwith Metal Oxides Reduction","volume":"192","author":"Villatoro-Monzo\u0301n","year":"2008","journal-title":"Water. Air. Soil\nPollut."},{"issue":"12","key":"2026081212021241300_cit6","doi-asserted-by":"publisher","first-page":"7073","DOI":"10.1021\/acs.est.5b00715","article-title":"Biodegradation: Updating the Concepts\nof Control for Microbial Cleanup in Contaminated Aquifers","volume":"49","author":"Meckenstock","year":"2015","journal-title":"Environ. Sci. 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