TINF2 is a major susceptibility gene in Danish patients with multiple primary melanoma.

TINF2 encodes the TINF2 protein, which is a subunit in the shelterin complex critical for telomere regulation. Three recent studies have associated six truncating germline variants in TINF2 that have previously been associated with a cancer predisposition syndrome (CPS) caused by elongation of the telomeres. This has added TINF2 to the long telomere syndrome genes, together with other telomere maintenance genes such as ACD, POT1, TERF2IP, and TERT. We report a clinical study of 102 Danish patients with multiple primary melanoma (MPM) in which a germline truncating variant in TINF2 (p.(Arg265Ter)) was identified in four unrelated participants. The telomere lengths of three variant carriers were >90% percentile. In a routine diagnostic setting, the variant was identified in two more families, including an additional MPM patient and monozygotic twins with thyroid cancer and other cancer types. A total of 10 individuals from six independent families were confirmed carriers, all with cancer history, predominantly melanoma. Our findings suggest a major role of TINF2 in Danish patients with MPM. In addition to melanoma, other cancers in the six families include thyroid, renal, breast, and sarcoma, supporting a CPS in which melanoma, thyroid cancer, and sarcoma predominate. Further studies are needed to establish the full spectrum of associated cancer types and characterize lifetime cancer risk in carriers.

Effect of oxic and anoxic conditions on intracellular storage of polyhydroxyalkanoate and polyphosphate in Magnetospirillum magneticum strain AMB-1.

Magnetotactic bacteria (MTB) are microorganisms widely inhabiting the oxic-anoxic interface of aquatic environments. Beside biomineralizing magnetic nanocrystals, MTBs are able to sequester various chemical elements (e.g., carbon and phosphorus) for the biogenesis of intracellular granules, like polyhydroxyalkanoate (PHA) and polyphosphate (polyP), making them potentially important in biogeochemical cycling. Yet, the environmental controls of intracellular storage of carbon and phosphorus in MTB remain poorly understood. Here, we investigated the influence of oxic, anoxic and transient oxic-anoxic conditions on intracellular storage of PHA and polyP in Magnetospirillum magneticum strain AMB-1. In the incubations with oxygen, transmission electron microscopy revealed intercellular granules highly rich in carbon and phosphorus, which were further interpreted as PHA and polyP based on chemical and Energy-Dispersive X-ray spectroscopy analysis. Oxygen had a strong effect on PHA and polyP storage in AMB-1 cells, as PHA and polyP granules accounted for up to 47 ± 23% and 5.1 ± 1.7% of the cytoplasmic space, respectively, during continuous oxic conditions, while granules disappeared in anoxic incubations. Poly 3-hydroxybutyrate (PHB) and poly 3-hydroxyvalerate (PHV) accounted for 0.59 ± 0.66% and 0.0033 ± 0.0088% of dry cell weight, respectively, in anoxic incubations, while the values increased by a factor of 7 and 37 after oxygen was introduced. The results highlight a tight link between oxygen, carbon and phosphorus metabolisms in MTB, where favorable oxic growth conditions can lead to metabolic induction of polyP and PHA granule biogenesis.

Corrigendum: Effect of oxic and anoxic conditions on intracellular storage of polyhydroxyalkanoate and polyphosphate in Magnetospirillum magneticum strain AMB-1.

[This corrects the article DOI: 10.3389/fmicb.2023.1203805.].

Electrochemical disinfection may increase the spread of antibiotic resistance genes by promoting conjugal plasmid transfer.

Current in the milliampere range can be used for electrochemical inactivation of bacteria. Yet, bacteria-including antibiotic resistant bacteria (ARB) may be subjected to sublethal conditions due to imperfect mixing or energy savings measures during electrochemical disinfection. It is not known whether such sublethal current intensities have the potential to stimulate plasmid transfer from ARB. In this study, conjugal transfer of plasmid pKJK5 was investigated between Pseudomonas putida strains under conditions reflecting electrochemical disinfection. Although the abundance of culturable and membrane-intact donor and recipient cells decreased with applied current (0-60 mA), both transconjugant density and transconjugant frequency increased. Both active chlorine and superoxide radicals were generated electrolytically, and ROS generation was induced. In addition, we detected significant over expression of a core oxidative stress defense gene (ahpCF) with current. Expression of selected conjugation related genes (traE, traI, trbJ, and trbL) also significantly correlated with current intensity. ROS accumulation, SOS response and subsequent derepression of conjugation are therefore the plausible consequence of sublethal current exposure. These findings suggest that sublethal intensities of current can enhance conjugal plasmid transfer, and that it is essential that conditions of electrochemical disinfection (applied voltage, current density, time and mixing) are carefully controlled to avoid conjugal ARG transmission.

Microbial bioremediation of produced water under different redox conditions in marine sediments.

The discharge of produced water from offshore oil platforms is an emerging concern due to its potential adverse effects on marine ecosystems. In this study, we investigated the feasibility and capability of using marine sediments for the bioremediation of produced water. We utilized a combination of porewater and solid phase analysis in a series of sediment batch incubations amended with produced water and synthetic produced water to determine the biodegradation of hydrocarbons under different redox conditions. Significant removal of benzene, toluene, ethylbenzene and xylene (BTEX) compounds was observed under different redox conditions, with biodegradation efficiencies of 93-97% in oxic incubations and 45-93% in anoxic incubations with nitrate, iron oxide or sulfate as the electron acceptor. Higher biodegradation rates of BTEX were obtained by incubations dominated by nitrate reduction (104-149 nmolC/cm3/d) and oxygen respiration (52-57 nmolC/cm3/d), followed by sulfate reduction (14-76 nmolC/cm3/d) and iron reduction (29-39 nmolC/cm3/d). Chemical fingerprint analysis showed that hydrocarbons were biodegraded to smaller alcohols/acids under oxic conditions compared to anoxic conditions with nitrate, indicating that the presence of oxygen facilitated a more complete biodegradation process. Toxicity of treated produced water to the marine copepod Acartia tonsa was reduced by half after sediment incubations with oxygen and nitrate. Our study emphasizes the possibility to use marine sediment as a biofilter for treating produced water at sea without extending the oil and gas platform or implementing a large-scale construction.

9p21.3 Microdeletion involving CDKN2A/2B in a young patient with multiple primary cancers and review of the literature.

Germline pathogenic variants in CDKN2A predispose to various cancers, including melanoma, pancreatic cancer, and neural system tumors, whereas CDKN2B variants are associated with renal cell carcinoma. A few case reports have described heterozygous germline deletions spanning both CDKN2A and CDKN2B associated with a cancer predisposition syndrome (CPS) that constitutes a risk of cancer beyond those associated with haploinsufficiency of each gene individually, indicating an additive effect or a contiguous gene deletion syndrome. We report a young woman with a de novo germline 9p21 microdeletion involving the CDKN2A/CDKN2B genes, who developed six primary cancers since childhood, including a very rare extraskeletal osteosarcoma (eOS) at the age of 8. To our knowledge this is the first report of eOS in a patient with CDKN2A/CDKN2B deletion.

A new self-understanding as chemo sufferer - a phenomenological study of everyday life with chemotherapy induced neuropathy among survivors after colorectal cancer.

PURPOSE: To explore the essential meaning of how sensory disturbances caused by Oxaliplatin influence self-understanding and freedom to live an everyday life among survivors after colorectal cancer. METHODS: Data was generated by means of a semi-structured individual interview with eight survivors after colorectal cancer who continued to experience chemotherapy-induced peripheral neuropathy at least one year after completing chemotherapy with Oxaliplatin. Data analysis was guided by existential phenomenology and descriptive life-world research. RESULTS: The essential meaning was structured by four constituents. 1) An unpleasant fluctuating sensation which is impossible to ignore, 2) Breaking through of noise and pain despite struggling to keep them at bay, 3) Continuously feeling ill despite being cured, and 4) Bodily constraints that impact self-understanding and limit enjoyment of life. CONCLUSION: The survivors used distraction to keep the sensory disturbances at bay but were forced to adapt to a new self-understanding as sufferers after chemotherapy despite being cured of their cancer disease. This way of being-in-the-world was understood by survivors, their families and healthcare professionals as a necessary price to pay to be alive. However, marked as sufferer after chemotherapy, the participants' everyday style of experience and life revealed as an ill health condition, which limited their ability to accomplish everyday activities as before and their freedom to realize their potential-the "I can".

Critical Assessment of MetaProteome Investigation (CAMPI): a multi-laboratory comparison of established workflows.

Metaproteomics has matured into a powerful tool to assess functional interactions in microbial communities. While many metaproteomic workflows are available, the impact of method choice on results remains unclear. Here, we carry out a community-driven, multi-laboratory comparison in metaproteomics: the critical assessment of metaproteome investigation study (CAMPI). Based on well-established workflows, we evaluate the effect of sample preparation, mass spectrometry, and bioinformatic analysis using two samples: a simplified, laboratory-assembled human intestinal model and a human fecal sample. We observe that variability at the peptide level is predominantly due to sample processing workflows, with a smaller contribution of bioinformatic pipelines. These peptide-level differences largely disappear at the protein group level. While differences are observed for predicted community composition, similar functional profiles are obtained across workflows. CAMPI demonstrates the robustness of present-day metaproteomics research, serves as a template for multi-laboratory studies in metaproteomics, and provides publicly available data sets for benchmarking future developments.

Evaluation of RNAlater as a Field-Compatible Preservation Method for Metaproteomic Analyses of Bacterium-Animal Symbioses.

Field studies are central to environmental microbiology and microbial ecology, because they enable studies of natural microbial communities. Metaproteomics, the study of protein abundances in microbial communities, allows investigators to study these communities "in situ," which requires protein preservation directly in the field because protein abundance patterns can change rapidly after sampling. Ideally, a protein preservative for field deployment works rapidly and preserves the whole proteome, is stable in long-term storage, is nonhazardous and easy to transport, and is available at low cost. Although these requirements might be met by several protein preservatives, an assessment of their suitability under field conditions when targeted for metaproteomic analyses is currently lacking. Here, we compared the protein preservation performance of flash freezing and the preservation solution RNAlater using the marine gutless oligochaete Olavius algarvensis and its symbiotic microbes as a test case. In addition, we evaluated long-term RNAlater storage after 1 day, 1 week, and 4 weeks at room temperature (22°C to 23°C). We evaluated protein preservation using one-dimensional liquid chromatography-tandem mass spectrometry. We found that RNAlater and flash freezing preserved proteins equally well in terms of total numbers of identified proteins and relative abundances of individual proteins, and none of the test time points was altered, compared to time zero. Moreover, we did not find biases against specific taxonomic groups or proteins with particular biochemical properties. Based on our metaproteomic data and the logistical requirements for field deployment, we recommend RNAlater for protein preservation of field-collected samples targeted for metaproteomic analyses. IMPORTANCE Metaproteomics, the large-scale identification and quantification of proteins from microbial communities, provide direct insights into the phenotypes of microorganisms on the molecular level. To ensure the integrity of the metaproteomic data, samples need to be preserved immediately after sampling to avoid changes in protein abundance patterns. In laboratory setups, samples for proteomic analyses are most commonly preserved by flash freezing; however, liquid nitrogen or dry ice is often unavailable at remote field locations, due to their hazardous nature and transport restrictions. Our study shows that RNAlater can serve as a low-hazard, easy-to-transport alternative to flash freezing for field preservation of samples for metaproteomic analyses. We show that RNAlater preserves the metaproteome equally well, compared to flash freezing, and protein abundance patterns remain stable during long-term storage for at least 4 weeks at room temperature.

A peculiar experience- everyday life with chronic sensory disturbances after oxaliplatin treatment for colorectal cancer - a phenomenological study.

Purpose: To deepen the understanding of how survivors' experience and give meaning to the embodied phenomenon of chronic sensory disturbances in everyday life after oxaliplatin treatment for colorectal cancer.Methods: Data was generated by means of a semi-structured interview guide and drawings with the aim to explore eight survivors' lifeworld experiences. Data was analyzed through a phenomenological approach.Results: The essential meaning of sensory disturbances emerged in two main themes and four sub-themes. Theme A: 'A peculiar experience that is difficult to logically understand' with the subthemes; 'An ambiguous perception in hands and feet' and 'Being alienated from one's own body'. Theme B: Losing touch with the world' with the subthemes: 'A lack of sensory contact with physical surfaces' and 'Breakdown of sensitivity in hands hampers fine motor skills and social contact'.Conclusion: Sensory disturbances contributed to an ambiguous and discordant perception of an alienated body that was difficult to describe and affected the ability to act and connect to things and other people. Metaphors and drawings were valuable as means to verbalize and illustrate the changed body perception where the 'I can' changed into 'I cannot'. To support the embodied connection to the world new usage patterns were required.

Role of Ammonia Oxidation in Organic Micropollutant Transformation during Wastewater Treatment: Insights from Molecular, Cellular, and Community Level Observations.

Organic micropollutants (OMPs) are a threat to aquatic environments, and wastewater treatment plants may act as a source or a barrier of OMPs entering the environment. Understanding the fate of OMPs in wastewater treatment processes is needed to establish efficient OMP removal strategies. Enhanced OMP biotransformation has been documented during biological nitrogen removal and has been attributed to the cometabolic activity of ammonia-oxidizing bacteria (AOB) and, specifically, to the ammonia monooxygenase (AMO) enzyme. Yet, the exact mechanisms of OMP biotransformation are often unknown. This critical review aims to fundamentally and quantitatively evaluate the role of ammonia oxidation in OMP biotransformation during wastewater treatment processes. OMPs can be transformed by AOB via direct and indirect enzymatic reactions: AMO directly transforms OMPs primarily via hydroxylation, while biologically produced reactive nitrogen species (hydroxylamine (NH2OH), nitrite (NO2-), and nitric oxide (NO)) can chemically transform OMPs through nitration, hydroxylation, and deamination and can contribute significantly to the observed OMP transformations. OMPs containing alkyl, aliphatic hydroxyl, ether, and sulfide functional groups as well as substituted aromatic rings and aromatic primary amines can be biotransformed by AMO, while OMPs containing alkyl groups, phenols, secondary amines, and aromatic primary amines can undergo abiotic transformations mediated by reactive nitrogen species. Higher OMP biotransformation efficiencies and rates are obtained in AOB-dominant microbial communities, especially in autotrophic reactors performing nitrification or nitritation, than in non-AOB-dominant microbial communities. The biotransformations of OMPs in wastewater treatment systems can often be linked to ammonium (NH4+) removal following two central lines of evidence: (i) Similar transformation products (i.e., hydroxylated, nitrated, and desaminated TPs) are detected in wastewater treatment systems as in AOB pure cultures. (ii) Consistency in OMP biotransformation (rbio, µmol/g VSS/d) to NH4+ removal (rNH4+, mol/g VSS/d) rate ratios (rbio/rNH4+) is observed for individual OMPs across different systems with similar rNH4+ and AOB abundances. In this review, we conclude that AOB are the main drivers of OMP biotransformation during wastewater treatment processes. The importance of biologically driven abiotic OMP transformation is quantitatively assessed, and functional groups susceptible to transformations by AMO and reactive nitrogen species are systematically classified. This critical review will improve the prediction of OMP transformation and facilitate the design of efficient OMP removal strategies during wastewater treatment.

Time to act-assessing variations in qPCR analyses in biological nitrogen removal with examples from partial nitritation/anammox systems.

Quantitative PCR (qPCR) is broadly used as the gold standard to quantify microbial community fractions in environmental microbiology and biotechnology. Benchmarking efforts to ensure the comparability of qPCR data for environmental bioprocesses are still scarce. Also, for partial nitritation/anammox (PN/A) systems systematic investigations are still missing, rendering meta-analysis of reported trends and generic insights potentially precarious. We report a baseline investigation of the variability of qPCR-based analyses for microbial communities applied to PN/A systems. Round-robin testing was performed for three PN/A biomass samples in six laboratories, using the respective in-house DNA extraction and qPCR protocols. The concentration of extracted DNA was significantly different between labs, ranged between 2.7 and 328 ng mg-1 wet biomass. The variability among the qPCR abundance data of different labs was very high (1-7 log fold) but differed for different target microbial guilds. DNA extraction caused maximum variation (3-7 log fold), followed by the primers (1-3 log fold). These insights will guide environmental scientists and engineers as well as treatment plant operators in the interpretation of qPCR data.

Involving patients and nurses in choosing between two validated questionnaires to identify chemotherapy-induced peripheral neuropathy before implementing in clinical practice-A qualitative study.

AIMS AND OBJECTIVES: To explore from a nurse and patient perspective what questionnaire-"Functional assessment of cancer treatment gynecological group neurotoxicity" or "Oxaliplatin-Associated Neuropathy Questionnaire"-best describes chemotherapy-induced peripheral neuropathy and its influence on everyday life in a comprehensive and meaningful way, prior to implementation in daily practice. BACKGROUND: Patients experience chemotherapy-induced peripheral neuropathy during and after chemotherapy for colorectal cancer with oxaliplatin. This neuropathy is difficult to describe for patients and to identify for nurses. To address the specific needs of patients and improve identification of neuropathy and its influence on everyday life, we wanted to implement a questionnaire in clinical practice. DESIGN: A phenomenological hermeneutic frame of reference was used. METHOD: Semi-structured interviews with 15 patients and two focus groups with eight cancer nurses were used for data collection. Data were organised and interpreted by content analytical steps in a hermeneutical process. COREQ checklist was used in reporting of the study. RESULTS: The analysis resulted in two main themes (a) "To dig deeper" with sub-themes "to identify the line between acceptable and nonacceptable chemotherapy-induced peripheral neuropathy," and "searching for a precise description." (b). "When everything is interrelated" with sub-themes "to be aware of different perspectives and understandings" and "recognise potential pitfalls." CONCLUSION: Involving patients and nurses in choosing between the two questionnaires revealed that neither alone was sufficient to describe the patients' experiences. Instead, it seems essential to implement both questionnaires, using the answers as a basis for a dialogue to address the patients' specific needs. RELEVANCE FOR CLINICAL PRACTICE: Using patients and nurses perspectives in a complementary way may provide a solid foundation before starting an implementation process in clinical practice. However, attention must be paid to potential barriers and facilitators as well as the fact that a successful implementing process requires leadership and information sharing.

The effect of pH on N2O production in intermittently-fed nitritation reactors.

The effect of pH on nitrous oxide (N2O) production rates was quantified in an intermittently-fed lab-scale sequencing batch reactor performing high-rate nitritation. N2O and other nitrogen (N) species (e.g. ammonium (NH4+), nitrite, hydroxylamine and nitric oxide) were monitored to identify in-cycle dynamics and determine N conversion rates at controlled pH set-points (6.5, 7, 7.5, 8 and 8.5). Operational conditions and microbial compositions remained similar during long-term reactor-scale pH campaigns. The specific ammonium removal rates and nitrite accumulation rates varied little with varying pH levels (p > 0.05). The specific net N2O production rates and net N2O yield of NH4+ removed (ΔN2O/ΔNH4+) increased up to seven-fold from pH 6.5 to 8, and decreased slightly with further pH increase to 8.5 (p < 0.05). Best-fit model simulations predicted nitrifier denitrification as the dominant N2O production pathway (≥87% of total net N2O production) at all examined pH. Our study highlights the effect of pH on biologically mediated N2O emissions in nitrogen removal systems and its importance in the design of N2O mitigation strategies.

Abiotic Nitrous Oxide (N2O) Production Is Strongly pH Dependent, but Contributes Little to Overall N2O Emissions in Biological Nitrogen Removal Systems.

Hydroxylamine (NH2OH) and nitrite (NO2-), intermediates during the nitritation process, can engage in chemical (abiotic) reactions that lead to nitrous oxide (N2O) generation. Here, we quantify the kinetics and stoichiometry of the relevant abiotic reactions in a series of batch tests under different and relevant conditions, including pH, absence/presence of oxygen, and reactant concentrations. The highest N2O production rates were measured from NH2OH reaction with HNO2, followed by HNO2 reduction by Fe2+, NH2OH oxidation by Fe3+, and finally NH2OH disproportionation plus oxidation by O2. Compared to other examined factors, pH had the strongest effect on N2O formation rates. Acidic pH enhanced N2O production from the reaction of NH2OH with HNO2 indicating that HNO2 instead of NO2- was the reactant. In departure from previous studies, we estimate that abiotic N2O production contributes little (< 3% of total N2O production) to total N2O emissions in typical nitritation reactor systems between pH 6.5 and 8. Abiotic contributions would only become important at acidic pH (≤ 5). In consideration of pH effects on both abiotic and biotic N2O production pathways, circumneutral pH set-points are suggested to minimize overall N2O emissions from nitritation systems.

[Chronic rhinosinusitis].

The aim of this review is to evaluate current guidelines for diagnosis, treatment and follow-up of patients with chronic rhinosinusitis. We discuss: 1) diagnostic criteria, 2) the use of supplementary tools like visual analogue scale, Sino-Nasal Outcome Test and Sniffin' Sticks, 3) the use of tests like allergy, serum IgE and biopsy and 4) comorbidity in relation to the unified airways concept. Furthermore, we evaluate: 1) initial treatment with topical steroids and nasal irrigation, 2) additional treatment options including surgery, systemic steroids and antibiotics and 3) treatment risks. Follow-up of patients is important for evaluating treatment effect and possible need of further treatment.

Reactor staging influences microbial community composition and diversity of denitrifying MBBRs- Implications on pharmaceutical removal.

The subdivision of biofilm reactor in two or more stages (i.e., reactor staging) represents an option for process optimisation of biological treatment. In our previous work, we showed that the gradient of influent organic substrate availability (induced by the staging) can influence the microbial activity (i.e., denitrification and pharmaceutical biotransformation kinetics) of a denitrifying three-stage Moving Bed Biofilm Reactor (MBBR) system. However, it is unclear whether staging and thus the long-term exposure to varying organic carbon type and loading influences the microbial community structure and diversity. In this study, we investigated biofilm structure and diversity in the three-stage MBBR system (S) compared to a single-stage configuration (U) and their relationship with microbial functions. Results from 16S rRNA amplicon libraries revealed a significantly higher microbial richness in the staged MBBR (at 99% sequence similarity) compared to single-stage MBBR. A more even and diverse microbial community was selected in the last stage of S (S3), likely due to exposure to carbon limitation during continuous-flow operation. A core of OTUs was shared in both systems, consisting of Burkholderiales, Xanthomonadales, Flavobacteriales and Sphingobacteriales, while MBBR staging selected for specific taxa (i.e., Candidate division WS6 and Deinococcales). Results from quantitative PCR (qPCR) showed that S3 exhibited the lowest abundance of 16S rRNA but the highest abundance of atypical nosZ, suggesting a selection of microbes with more diverse N-metabolism (i.e., incomplete denitrifiers) in the stage exposed to the lowest carbon availability. A positive correlation (p < 0.05) was observed between removal rate constants of several pharmaceuticals with abundance of relevant denitrifying genes, but not with biodiversity. Despite the previously suggested positive relationship between microbial diversity and functionality in macrobial and microbial ecosystems, this was not observed in the current study, indicating a need to further investigate structure-function relationships for denitrifying systems.

The pH dependency of N-converting enzymatic processes, pathways and microbes: effect on net N2 O production.

Nitrous oxide (N2 O) is emitted during microbiological nitrogen (N) conversion processes, when N2 O production exceeds N2 O consumption. The magnitude of N2 O production vs. consumption varies with pH and controlling net N2 O production might be feasible by choice of system pH. This article reviews how pH affects enzymes, pathways and microorganisms that are involved in N-conversions in water engineering applications. At a molecular level, pH affects activity of cofactors and structural elements of relevant enzymes by protonation or deprotonation of amino acid residues or solvent ligands, thus causing steric changes in catalytic sites or proton/electron transfer routes that alter the enzymes' overall activity. Augmenting molecular information with, e.g., nitritation or denitrification rates yields explanations of changes in net N2 O production with pH. Ammonia oxidizing bacteria are of highest relevance for N2 O production, while heterotrophic denitrifiers are relevant for N2 O consumption at pH > 7.5. Net N2 O production in N-cycling water engineering systems is predicted to display a 'bell-shaped' curve in the range of pH 6.0-9.0 with a maximum at pH 7.0-7.5. Net N2 O production at acidic pH is dominated by N2 O production, whereas N2 O consumption can outweigh production at alkaline pH. Thus, pH 8.0 may be a favourable pH set-point for water treatment applications regarding net N2 O production.

Low nitrous oxide production through nitrifier-denitrification in intermittent-feed high-rate nitritation reactors.

Nitrous oxide (N2O) production from autotrophic nitrogen conversion processes, especially nitritation systems, can be significant, requires understanding and calls for mitigation. In this study, the rates and pathways of N2O production were quantified in two lab-scale sequencing batch reactors operated with intermittent feeding and demonstrating long-term and high-rate nitritation. The resulting reactor biomass was highly enriched in ammonia-oxidizing bacteria, and converted ∼93 ± 14% of the oxidized ammonium to nitrite. The low DO set-point combined with intermittent feeding was sufficient to maintain high nitritation efficiency and high nitritation rates at 20-26 °C over a period of ∼300 days. Even at the high nitritation efficiencies, net N2O production was low (∼2% of the oxidized ammonium). Net N2O production rates transiently increased with a rise in pH after each feeding, suggesting a potential effect of pH on N2O production. In situ application of 15N labeled substrates revealed nitrifier denitrification as the dominant pathway of N2O production. Our study highlights operational conditions that minimize N2O emission from two-stage autotrophic nitrogen removal systems.

Pathways and Controls of N2O Production in Nitritation-Anammox Biomass.

Nitrous oxide (N2O) is an unwanted byproduct during biological nitrogen removal processes in wastewater. To establish strategies for N2O mitigation, a better understanding of production mechanisms and their controls is required. A novel stable isotope labeling approach using 15N and 18O was applied to investigate pathways and controls of N2O production by biomass taken from a full-scale nitritation-anammox reactor. The experiments showed that heterotrophic denitrification was a negligible source of N2O under oxic conditions (≥0.2 mg O2 L-1). Both hydroxylamine oxidation and nitrifier denitrification contributed substantially to N2O accumulation across a wide range of conditions with varying concentrations of O2, NH4+, and NO2-. The O2 concentration exerted the strongest control on net N2O production with both production pathways stimulated by low O2, independent of NO2- concentrations. The stimulation of N2O production from hydroxylamine oxidation at low O2 was unexpected and suggests that more than one enzymatic pathway may be involved in this process. N2O production by hydroxylamine oxidation was further stimulated by NH4+, whereas nitrifier denitrification at low O2 levels was stimulated by NO2- at levels as low as 0.2 mM. Our study shows that 15N and 18O isotope labeling is a useful approach for direct quantification of N2O production pathways applicable to diverse environments.