PMID- 32607646 OWN - NLM STAT- MEDLINE DCOM- 20210416 LR - 20210416 IS - 1432-0614 (Electronic) IS - 0175-7598 (Print) IS - 0175-7598 (Linking) VI - 104 IP - 16 DP - 2020 Aug TI - Diversity, enrichment, and genomic potential of anaerobic methane- and ammonium-oxidizing microorganisms from a brewery wastewater treatment plant. PG - 7201-7212 LID - 10.1007/s00253-020-10748-z [doi] AB - Anaerobic wastewater treatment offers several advantages; however, the effluent of anaerobic digesters still contains high levels of ammonium and dissolved methane that need to be removed before these effluents can be discharged to surface waters. The simultaneous anaerobic removal of methane and ammonium by denitrifying (N-damo) methanotrophs in combination with anaerobic ammonium-oxidizing (anammox) bacteria could be a potential solution to this challenge. After a molecular survey of a wastewater plant treating brewery effluent, indicating the presence of both N-damo and anammox bacteria, we started an anaerobic bioreactor with a continuous supply of methane, ammonium, and nitrite to enrich these anaerobic microorganisms. After 14 months of operation, a stable enrichment culture containing two types of 'Candidatus Methylomirabilis oxyfera' bacteria and two strains of 'Ca. Brocadia'-like anammox bacteria was achieved. In this community, anammox bacteria converted 80% of the nitrite with ammonium, while 'Ca. Methylomirabilis' contributed to 20% of the nitrite consumption. The analysis of metagenomic 16S rRNA reads and fluorescence in situ hybridization (FISH) correlated well and showed that, after 14 months, 'Ca. Methylomirabilis' and anammox bacteria constituted approximately 30 and 20% of the total microbial community. In addition, a substantial part (10%) of the community consisted of Phycisphaera-related planctomycetes. Assembly and binning of the metagenomic sequences resulted in high-quality draft genome of two 'Ca. Methylomirabilis' species containing the marker genes pmoCAB, xoxF, and nirS and putative NO dismutase genes. The anammox draft genomes most closely related to 'Ca. Brocadia fulgida' included the marker genes hzsABC, hao, and hdh. Whole-reactor and batch anaerobic activity measurements with methane, ammonium, nitrite, and nitrate revealed an average anaerobic methane oxidation rate of 0.12 mmol h(-1) L(-1) and ammonium oxidation rate of 0.5 mmol h(-1) L(-1). Together, this study describes the enrichment and draft genomes of anaerobic methanotrophs from a brewery wastewater treatment plant, where these organisms together with anammox bacteria can contribute significantly to the removal of methane and ammonium in a more sustainable way. KEY POINTS: * An enrichment culture containing both N-damo and anammox bacteria was obtained. * Simultaneous consumption of ammonia, nitrite, and methane under anoxic conditions. * In-depth metagenomic biodiversity analysis of inoculum and enrichment culture. FAU - Stultiens, Karin AU - Stultiens K AD - Department of Microbiology, IWWR, Radboud University Nijmegen, Nijmegen, The Netherlands. FAU - van Kessel, Maartje A H J AU - van Kessel MAHJ AD - Department of Microbiology, IWWR, Radboud University Nijmegen, Nijmegen, The Netherlands. AD - Soehngen Institute of Anaerobic Microbiology, Nijmegen, The Netherlands. FAU - Frank, Jeroen AU - Frank J AD - Department of Microbiology, IWWR, Radboud University Nijmegen, Nijmegen, The Netherlands. FAU - Fischer, Peter AU - Fischer P AD - Department of Microbiology, IWWR, Radboud University Nijmegen, Nijmegen, The Netherlands. AD - Soehngen Institute of Anaerobic Microbiology, Nijmegen, The Netherlands. AD - Laboratory of Microbiology, Wageningen University & Research, Wageningen, The Netherlands. FAU - Pelzer, Chris AU - Pelzer C AD - Department of Microbiology, IWWR, Radboud University Nijmegen, Nijmegen, The Netherlands. FAU - van Alen, Theo A AU - van Alen TA AD - Department of Microbiology, IWWR, Radboud University Nijmegen, Nijmegen, The Netherlands. FAU - Kartal, Boran AU - Kartal B AD - Department of Microbiology, IWWR, Radboud University Nijmegen, Nijmegen, The Netherlands. AD - MPI Bremen, Bremen, Germany. FAU - Op den Camp, Huub J M AU - Op den Camp HJM AUID- ORCID: 0000-0003-1990-9030 AD - Department of Microbiology, IWWR, Radboud University Nijmegen, Nijmegen, The Netherlands. h.opdencamp@science.ru.nl. FAU - Jetten, Mike S M AU - Jetten MSM AD - Department of Microbiology, IWWR, Radboud University Nijmegen, Nijmegen, The Netherlands. AD - Soehngen Institute of Anaerobic Microbiology, Nijmegen, The Netherlands. LA - eng GR - 669371/H2020 European Research Council/ GR - 6400422/H2020 European Research Council/ GR - 339880/H2020 European Research Council/ GR - 13146/Stichting voor de Technische Wetenschappen/ GR - 016.veni.192.062/Nederlandse Organisatie voor Wetenschappelijk Onderzoek/ GR - 024002002/Nederlandse Organisatie voor Wetenschappelijk Onderzoek/ PT - Journal Article DEP - 20200630 PL - Germany TA - Appl Microbiol Biotechnol JT - Applied microbiology and biotechnology JID - 8406612 RN - 0 (Ammonium Compounds) RN - 0 (RNA, Ribosomal, 16S) RN - OP0UW79H66 (Methane) SB - IM MH - Ammonium Compounds/*metabolism MH - Anaerobiosis MH - Bacteria/*classification/metabolism MH - *Biodiversity MH - Bioreactors/*microbiology MH - Metagenomics MH - Methane/*metabolism MH - Oxidation-Reduction MH - RNA, Ribosomal, 16S/genetics MH - Water Purification PMC - PMC7374466 OTO - NOTNLM OT - Anaerobic oxidation of methane OT - Anammox OT - Hydrazine synthase OT - Metagenome OT - Nitrite OT - pmoA OT - 'Ca. Brocadia' OT - 'Ca. Methanoperedens nitroreducens' OT - 'Ca. Methylomirabilis' bacteria COIS- The authors declare that they have no conflict of interest. EDAT- 2020/07/02 06:00 MHDA- 2021/04/17 06:00 PMCR- 2020/06/30 CRDT- 2020/07/02 06:00 PHST- 2020/05/04 00:00 [received] PHST- 2020/06/16 00:00 [accepted] PHST- 2020/06/11 00:00 [revised] PHST- 2020/07/02 06:00 [pubmed] PHST- 2021/04/17 06:00 [medline] PHST- 2020/07/02 06:00 [entrez] PHST- 2020/06/30 00:00 [pmc-release] AID - 10.1007/s00253-020-10748-z [pii] AID - 10748 [pii] AID - 10.1007/s00253-020-10748-z [doi] PST - ppublish SO - Appl Microbiol Biotechnol. 2020 Aug;104(16):7201-7212. doi: 10.1007/s00253-020-10748-z. Epub 2020 Jun 30.