{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,5,19]],"date-time":"2026-05-19T13:10:35Z","timestamp":1779196235250,"version":"3.51.4"},"reference-count":39,"publisher":"American Society for Microbiology","issue":"1","license":[{"start":{"date-parts":[[2004,1,1]],"date-time":"2004-01-01T00:00:00Z","timestamp":1072915200000},"content-version":"tdm","delay-in-days":0,"URL":"https:\/\/journals.asm.org\/non-commercial-tdm-license"}],"content-domain":{"domain":["journals.asm.org"],"crossmark-restriction":true},"short-container-title":["Appl Environ Microbiol"],"published-print":{"date-parts":[[2004,1]]},"abstract":"<jats:title>ABSTRACT<\/jats:title>\n          <jats:p>\n            A denitrifying microbial consortium was enriched in an anoxically operated, methanol-fed sequencing batch reactor (SBR) fed with a mineral salts medium containing methanol as the sole carbon source and nitrate as the electron acceptor. The SBR was inoculated with sludge from a biological nutrient removal activated sludge plant exhibiting good denitrification. The SBR denitrification rate improved from less than 0.02 mg of NO\n            <jats:sub>3<\/jats:sub>\n            <jats:sup>\u2212<\/jats:sup>\n            -N mg of mixed-liquor volatile suspended solids (MLVSS)\n            <jats:sup>\u22121<\/jats:sup>\n            h\n            <jats:sup>\u22121<\/jats:sup>\n            to a steady-state value of 0.06 mg of NO\n            <jats:sub>3<\/jats:sub>\n            <jats:sup>\u2212<\/jats:sup>\n            -N mg of MLVSS\n            <jats:sup>\u22121<\/jats:sup>\n            h\n            <jats:sup>\u22121<\/jats:sup>\n            over a 7-month operational period. At this time, the enriched microbial community was subjected to stable-isotope probing (SIP) with [\n            <jats:sup>13<\/jats:sup>\n            C]methanol to biomark the DNA of the denitrifiers. The extracted [\n            <jats:sup>13<\/jats:sup>\n            C]DNA and [\n            <jats:sup>12<\/jats:sup>\n            C]DNA from the SIP experiment were separately subjected to full-cycle rRNA analysis. The dominant 16S rRNA gene phylotype (group A clones) in the [\n            <jats:sup>13<\/jats:sup>\n            C]DNA clone library was closely related to those of the obligate methylotrophs\n            <jats:italic>Methylobacillus<\/jats:italic>\n            and\n            <jats:italic>Methylophilus<\/jats:italic>\n            in the order\n            <jats:italic>Methylophilales<\/jats:italic>\n            of the\n            <jats:italic>Betaproteobacteria<\/jats:italic>\n            (96 to 97% sequence identities), while the most abundant clone groups in the [\n            <jats:sup>12<\/jats:sup>\n            C]DNA clone library mostly belonged to the family\n            <jats:italic>Saprospiraceae<\/jats:italic>\n            in the\n            <jats:italic>Bacteroidetes<\/jats:italic>\n            phylum. Oligonucleotide probes for use in fluorescence in situ hybridization (FISH) were designed to specifically target the group A clones and\n            <jats:italic>Methylophilales<\/jats:italic>\n            (probes DEN67 and MET1216, respectively) and the\n            <jats:italic>Saprospiraceae<\/jats:italic>\n            clones (probe SAP553). Application of these probes to the SBR biomass over the enrichment period demonstrated a strong correlation between the level of SBR denitrification and relative abundance of DEN67-targeted bacteria in the SBR community. By contrast, there was no correlation between the denitrification rate and the relative abundances of the well-known denitrifying genera\n            <jats:italic>Hyphomicrobium<\/jats:italic>\n            and\n            <jats:italic>Paracoccus<\/jats:italic>\n            or the\n            <jats:italic>Saprospiraceae<\/jats:italic>\n            clones visualized by FISH in the SBR biomass. FISH combined with microautoradiography independently confirmed that the DEN67-targeted cells were the dominant bacterial group capable of anoxic [\n            <jats:sup>14<\/jats:sup>\n            C]methanol uptake in the enriched biomass. The well-known denitrification lag period in the methanol-fed SBR was shown to coincide with a lag phase in growth of the DEN67-targeted denitrifying population. We conclude that\n            <jats:italic>Methylophilales<\/jats:italic>\n            bacteria are the dominant denitrifiers in our SBR system and likely are important denitrifiers in full-scale methanol-fed denitrifying sludges.\n          <\/jats:p>","DOI":"10.1128\/aem.70.1.588-596.2004","type":"journal-article","created":{"date-parts":[[2004,1,7]],"date-time":"2004-01-07T21:02:54Z","timestamp":1073509374000},"page":"588-596","update-policy":"https:\/\/doi.org\/10.1128\/asmj-crossmark-policy-page","source":"Crossref","is-referenced-by-count":193,"title":["Use of Stable-Isotope Probing, Full-Cycle rRNA Analysis, and Fluorescence In Situ Hybridization-Microautoradiography To Study a Methanol-Fed Denitrifying Microbial Community"],"prefix":"10.1128","volume":"70","author":[{"given":"Maneesha P.","family":"Ginige","sequence":"first","affiliation":[{"name":"Advanced Wastewater Management Centre, The University of Queensland, St. Lucia 4072, Queensland, Australia"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Philip","family":"Hugenholtz","sequence":"additional","affiliation":[{"name":"Department of Environmental Science, Policy, and Management, University of California, Berkeley, California 94720-3110"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Holger","family":"Daims","sequence":"additional","affiliation":[{"name":"Abteilung fu\u0308r Mikrobielle O\u0308kologie, Institut fu\u0308r O\u0308kologie und Naturschutz, Universita\u0308t Wien, 1090 Vienna, Austria"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Michael","family":"Wagner","sequence":"additional","affiliation":[{"name":"Abteilung fu\u0308r Mikrobielle O\u0308kologie, Institut fu\u0308r O\u0308kologie und Naturschutz, Universita\u0308t Wien, 1090 Vienna, Austria"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Ju\u0308rg","family":"Keller","sequence":"additional","affiliation":[{"name":"Advanced Wastewater Management Centre, The University of Queensland, St. Lucia 4072, Queensland, Australia"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Linda L.","family":"Blackall","sequence":"additional","affiliation":[{"name":"Advanced Wastewater Management Centre, The University of Queensland, St. Lucia 4072, Queensland, Australia"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"235","reference":[{"key":"e_1_3_2_2_2","doi-asserted-by":"publisher","DOI":"10.1016\/S0022-2836(05)80360-2"},{"key":"e_1_3_2_3_2","doi-asserted-by":"publisher","DOI":"10.1128\/AEM.56.6.1919-1925.1990"},{"key":"e_1_3_2_4_2","doi-asserted-by":"publisher","DOI":"10.1128\/mr.59.1.143-169.1995"},{"key":"e_1_3_2_5_2","unstructured":"American Public Health Association. 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