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Advanced understanding of autotrophic nitrogen removal and associated N2O emissions in mixed nitritation-anammox systems through combined stable ISOtopic and MOLecular constraints (ISOMOL)

Applicant Lehmann Moritz
Number 170876
Funding scheme Sinergia
Research institution Institut für Umweltgeowissenschaften Universität Basel
Institution of higher education University of Basel - BS
Main discipline Interdisciplinary
Start/End 01.09.2017 - 31.08.2021
Approved amount 1'484'426.00
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All Disciplines (6)

Other disciplines of Engineering Sciences
Climatology. Atmospherical Chemistry, Aeronomy
Other disciplines of Environmental Sciences

Keywords (10)

microbial community; dissolved inorganic nitrogen; ecological stability; wastewater treatment; autotrophic N removal; N2O; stable isotopes; greenhouse gas; molecular biology; anammox

Lay Summary (German)

Das ISOMOL-Projekt zielt darauf ab, Isotopenmessungen und molekular-biologische Untersuchungen zu kombinieren, um den Stickstoff-Stoffwechsel und Isotopeneffekte in Anammox Misch- und Reinkulturen besser zu verstehen und Mechanismen der anammox-assoziierten N2O-Produktion aufzuklären sowie optimale Bedingungen für eine effiziente und stabile Stickstoffelimination bei der Abwasserbehandlung zu finden.
Lay summary

Das partielle Nitritation/Anammox Verfahren (PN/A) ist ein neues Verfahren, das eine energie- und kosteneffizientere Entfernung von reaktiven Stickstoffverbindungen wie Ammonium, Nitrat und Nitrit (Stickstoffdünger) in Kläranlagen ermöglicht. Das PN/A Verfahren steht heute jedoch immer noch auf dem Prüfstand, da für eine optimale Faulwasserbehandlung eine hohe Prozessstabilität, kombiniert mit optimaler Eliminationseffizienz und minimalen Treibhausgasemissionen verlangt werden. Anammox, die anaerobe Oxidation von Ammonium mit Nitrit durch spezielle Mikroorganismen, spielt auch in natürlichen aquatischen Systemen (Ozean, Seen, Grundwasser) eine wichtige Rolle. Die genauen Kontrollmechanismen, die Ökologie der beteiligten Mikroorganismen und die Umweltrelevanz der vom Anammox-Prozess freigesetzten Treibhausgase (z.B. N2O) sind jedoch noch weitgehend unbekannt.

Das ISOMOL-Projekt zielt darauf ab, Isotopenmessungen und molekular-biologische Untersuchungen zu kombinieren, um einerseits den Stickstoff-Stoffwechsel und Isotopeneffekte in Anammox Misch- und Reinkulturen besser zu verstehen und Mechanismen der anammox-assoziierten N2O-Produktion aufzuklären. Andererseits sollen diese Methoden, in Kombination mit Modellierungen, verfahrenstechnisch genutzt werden, um optimale Bedingungen für eine effiziente und stabile Stickstoffelimination zu finden. Die gewonnen Erkenntnisse werden nicht nur von grosser Wichtigkeit für die Optimierung des PN/A Verfahrens bei der Abwasserbehandlung sein, sondern auch zu einem besseren Verständnis der Regulationsmechanismen der mikrobiellen Stickstoffelimination durch Anammox in natürlichen Ökosystemen beitragen. Ausserdem sollen im Rahmen des ISOMOL Projekts die Nutzbarkeit von Isotopenmessungen für die Identifizierung und Quantifizierung von Stickstoffumsatzreaktionen im allgemeinen verbessert werden.

Direct link to Lay Summary Last update: 26.01.2017

Responsible applicant and co-applicants


Project partner


First real-time isotopic characterisation of N2O from chemodenitrification
Wei Jing (2019), First real-time isotopic characterisation of N2O from chemodenitrification, in Geochimica et Cosmochimica Acta, 15, 17-32.


Group / person Country
Types of collaboration
Toykyo Institute of Technology (Prof. Naohiro Yoshida) Japan (Asia)
- in-depth/constructive exchanges on approaches, methods or results
- Publication
Bundesanstalt für Gewässerkunde, Koblenz (Prof. Thomas Ternes) Germany (Europe)
- in-depth/constructive exchanges on approaches, methods or results
- Publication
University if Texas, El Paso (Prof. Benjamin Brunner) United States of America (North America)
- in-depth/constructive exchanges on approaches, methods or results
- Publication
- Exchange of personnel

Scientific events

Active participation

Title Type of contribution Title of article or contribution Date Place Persons involved
Goldschmidt Conference Talk given at a conference Stable isotope approaches to characterizing anammox and associated microbial processes in engineered and environmental systems 18.08.2019 Barcelona, Spain Lehmann Moritz; Magyar Paul;
Isotopes Talk given at a conference Quantifying N2O production pathways of waste water treatment plants using 15N tracer 07.07.2019 Munich, Germany Wei Jing;
Agro2019 - 10th IWA International Symposium on Waste Management Problems in Agro–Industries Talk given at a conference Identification of main N2O production pathways of a novel energy-neutral nitrogen-removal process for wastewater treatment 10.06.2019 Rhodes, Greece Wei Jing;
Meeting of the European Geophysical Union Poster First real-time isotopic characterization of N2O from chemodenitrification. 08.04.2019 Vienna, Austria Wei Jing;
DASIM19: “Tracing Denitrification” Poster Identification of Main N2O Production Pathways of a Novel Energy Neutral Nitrogen-Removal Process for Waste Water Treatment 12.03.2019 Giessen, Germany Wei Jing;
Swiss Microbial Ecology Meeting Poster Resistance, resilience and ecological stability of anammox consortia to transient oxygen disturbances 30.01.2019 Lausanne, Switzerland Bürgmann Helmut; Niederdorfer Robert Thomas;
Goldschmidt Conference Individual talk Nitrogen and oxygen isotope effects associated with a combined partial nitritation-anammox wastewater treatment process 12.08.2018 Boston, United States of America Magyar Paul;
ISME Conference Talk given at a conference Resistance, resilience and ecological stability of anammox biofilms to transient oxygen disturbances 12.08.2018 Leipzig, Germany Bürgmann Helmut; Niederdorfer Robert Thomas;

Associated projects

Number Title Start Funding scheme
169142 The Regulation of Nitrogen Turnover in Lakes 01.03.2017 Project funding (Div. I-III)
153055 Seasonal Dynamics of Coupled Nitrogen, Sulfur, and Carbon Cycling in Redox Transition Zones of Lake Lugano 01.11.2014 Project funding (Div. I-III)
150237 N2O from the Swiss midlands - regional sources and hot spots 01.06.2014 Project funding (Div. I-III)
163075 Assessment of the global N2O budget based on seasonal and long-term isotope measurements at Jungfraujoch and the Cape Grim Air Archive 01.12.2016 Project funding (Div. I-III)
129491 Nitrogen elimination pathways and associated isotope effects in Swiss eutrophic Lake Lugano 01.04.2010 Project funding (Div. I-III)
147106 Isotopic constraints on seasonal N2O dynamics in marine and lacustrine environments 01.04.2013 Project funding (Div. I-III)


Autotrophic nitrogen (N) removal by anaerobic ammonium oxidation (anammox) is an important mechanism of fixed N elimination, both in engineered and natural systems. In wastewater treatment plants it may permit operation under energy autarky and with a better carbon footprint. However, its process control and engineering is still under development: An optimized removal process using nitritation-anammox systems must combine stable operation, high N removal efficiency and minimized greenhouse gas emissions. Yet the biogeochemical and microbiological controls on the production of N2O in wastewater treatment are poorly understood. Similarly, the links between system stability, activity, and microbial population shifts are not well constrained. Equally important, in natural aquatic environments, the anammox process probably plays a much greater role than it was assumed for decades, challenging the traditional perception of the natural N-cycle. The questions regarding the prime pathways of fixed N loss in natural environments (i.e., denitrification versus anammox), the controls on the balance between fixed N loss and N2O production, and the associated microbial population dynamics are important rationales of current research. In this applied and environmental context, an important focus of the project will be on the use of stable isotope measurements and microbiological analyses to understand and identify N metabolism in pure and mixed anammox cultures, to assess the controls on N2O production in nitritation-anammox systems, and to verify ties between microbial dynamics, N-turnover and optimal process control.The collaborative and highly interdisciplinary project “IsoMol”, led by M. Lehmann (Unibas), seeks to understand the functioning of mixed microbial populations featuring alternative biogeochemical pathways and to characterize its dependence on environmental conditions and microbial composition. Each subproject will provide important information to improve our understanding of anammox on multiple levels (process engineering, isotope dynamics, microbiology). Subproject 1, led by A. Joss (EAWAG) has a strong applied component, which includes the set-up of enriched and mixed culture nitritation-anammox reactors at the laboratory and pilot scales. It aims at understanding both process controls and N2O emission in these systems (e.g., oxygen supply and temperature), and to evaluate the benefits of anammox-based wastewater treatment with respect to conventional approaches. Sub-projects 2 and 3, each with completely different analytical approaches, will develop and test stable isotope methods to understand and fingerprint N metabolism in pure and mixed cultures. Ammonium, nitrite and nitrate N and oxygen (O) isotope fractionation will be studied by M. Lehmann (Unibas), with a particular focus on the isotopic effect of anaerobic nitrate formation by anammox, as well as the potential of N and O isotope measurements to diagnose chemical transformations and reaction rates for different inorganic N substrates in general. Gas phase isotopic composition studies, led by J. Mohn (Empa), will identify the main processes leading to the emission of N2O across the range of typical operating conditions - in particular, with respect to a possible new pathway associated with the anammox metabolism. Subproject 4, led by H. Bürgmann (Eawag), will make use of cutting-edge techniques in microbiology and genetics, specifically metagenomics and metatranscriptomics, to understand the structure and function of mixed-culture anammox consortia in response to key external variables, and the mechanisms that control stability. It will apply concepts of ecological stability to the anammox process, aiming for fundamental advances in our understanding of what drives resistance and resilience of the involved microbial communities and processes. Energy-neutral municipal wastewater treatment is feasible with autotrophic N removal, but requires an optimized process design and control based on fundamental understanding of bacterial behavior and interactions. N (and O) isotope measurements in different N-species hold a strong potential to discriminate between alternative biochemical pathways and quantify reaction rates, to assess process stability and to understand the conditions that are most conducive to the emission of N2O. Similarly, the engineered nature of the community provides a unique level of control over what is nonetheless a complex real-life microbial community, and results will be immediately applicable to develop improvements for process engineering. In addition, the proposed research will provide the first in-depth transcriptomic analysis of anammox enrichments, and will significantly deepen our understanding of the physiological and regulatory differences between anammox strains. Our research may also contribute to the discovery and understanding of new modes of N2O production directly or indirectly related to anammox. Most importantly, expected break-through findings will not only be relevant for optimizing wastewater treatment, but they will also advance our understanding of N transformations in general, both at the molecular and ecosystem levels.