Coupled Nitrification Denitrification Research Articles

The impacts of film mulching and ridging on N2O emissions, relevant functional genes, and microbial communities in rain-fed potato fields

Rain-fed potato (Solanum tuberosum) fields in drylands significantly contribute to nitrous oxide (N2O) emissions, making them an important focus of agricultural greenhouse gas research. Film mulching and ridging are key agricultural methods in potato cultivation. Investigating the impact of these methods on N2O emissions, nitrifying/denitrifying functional genes, and microbial communities can provide a theoretical basis for soil emission reduction and more sustainable dryland agriculture. We examine the effects of flat tillage with mulching, ridge tillage with mulching, flat tillage without mulching, and ridge tillage without mulching, on potato fields under natural rainfall conditions in Wuchuan County, China. N2O emission fluxes were monitored using a static (dark) chamber and gas chromatography. Real-time quantitative PCR (q-PCR) was used to quantify abundances of nitrifying and denitrifying bacteria related to N2O emissions at various potato-growth stages. Illumina high-throughput sequencing was used to investigate microbial community structure by targeting 16S rRNA genes; related soil elements (soil temperatures and moisture) are analyzed. Mulching and ridging indirectly influence N2O emissions, nitrifying/denitrifying functional gene copy numbers, and microbial community structure by altering soil temperature and moisture. Cumulative N2O emissions and emission intensity were both consistently higher in ridge tillage with mulching during the potato-growing period. Ammonia-oxidizing archaea are the main microorganisms that control N2O emissions, with nitrification-coupled denitrification also being an important mechanism contributing to high N2O emissions during soil dry–wet cycles. Increased soil temperature and moisture elevated N2O emissions and functional gene copy numbers. The combination of mulching and ridging effectively uses the characteristics of both practices, making Nitrospira the dominant genus, and significantly increases N2O emissions.

Ocean acidification offsets the effect of warming on sediment denitrification and associated nitrous oxide production

AbstractRates of denitrification and associated nitrous oxide (N2O) production are expected to increase with global warming, leading to positive climate feedback. However, previous studies have not considered the combined effect of ocean acidification (OA, pCO2 ~ 900 µatm) and warming on denitrification rates and N2O production. Here we used a series of whole core incubation studies to assess the combined impact of warming and OA on estuarine sediment denitrification rates and N2O production. Strong warming (+5 °C over mean in situ conditions) increased N2O production by ~4.2 µmol-N m−2 d−1 and denitrification by ~43 µmol-N m−2 d−1, fuelled by water column nitrate (Dw), but decreased rates of nitrification-coupled denitrification in the sediment (Dn) by ~82 µmol-N m−2 d−1. While Dn was not affected by OA, Dw decreased significantly by 51 µmol-N m−2 d−1 when OA was coupled with warmer temperatures. We estimate that OA may offset the increase in estuarine sediment denitrification and N2O production expected from warming alone by up to 64% and reduce a potential positive climate feedback loop by inhibiting denitrification pathways.

Plastic particles affect N2O release via altering core microbial metabolisms in constructed wetlands

Constructed wetlands (CWs) have been proven to effectively immobilize plastic particles. However, little is known about the differences in the impact of varying sized plastic particles on nitrous oxide (N2O) release, as well as the intervention mechanisms in CWs. Here, we built a lab-scale wetland model and introduced plastic particles of macro-, micro-, and nano-size at 100 μg/L for 370 days. The results showed that plastic particles of all sizes reduced N2O release in CWs, with the degrees being the strongest for the Nano group, followed by Micro and Macro groups. Meanwhile, 15N- and 18O-tracing experiment revealed that the ammoxidation process contributed the most N2O production, followed by denitrification. While for every N2O-releasing process, the contributing proportion of N2O in nitrification-coupled denitrification were most significantly cut down under exposing to macro-sized plastics and had an obvious increase in nitrifier denitrification in all groups, respectively. Finally, we revealed the three mechanism pathways of N2O release reduction with macro-, micro-, and nano-sized plastics by impacting carbon assimilation (RubisCO activity), ammonia oxidation (gene amo abundance and HAO activity), and N-ion transmembrane and reductase activities, respectively. Our findings thus provided novel insights into the potential effects of plastic particles in CWs as an eco-technology.

Non-negligible N2O emission hotspots: Rivers impacted by ion-adsorption rare earth mining

Rare earth mining causes severe riverine nitrogen pollution, but its effect on nitrous oxide (N2O) emissions and the associated nitrogen transformation processes remain unclear. Here, we characterized N2O fluxes from China's largest ion-adsorption rare earth mining watershed and elucidated the mechanisms that drove N2O production and consumption using advanced isotope mapping and molecular biology techniques. Compared to the undisturbed river, the mining-affected river exhibited higher N2O fluxes (7.96 ± 10.18 mmol m−2d−1 vs. 2.88 ± 8.27 mmol m−2d−1, P = 0.002), confirming that mining-affected rivers are N2O emission hotspots. Flux variations scaled with high nitrogen supply (resulting from mining activities), and were mainly attributed to changes in water chemistry (i.e., pH, and metal concentrations), sediment property (i.e., particle size), and hydrogeomorphic factors (e.g., river order and slope). Coupled nitrification-denitrification and N2O reduction were the dominant processes controlling the N2O dynamics. Of these, the contribution of incomplete denitrification to N2O production was greater than that of nitrification, especially in the heavily mining-affected reaches. Co-occurrence network analysis identified Thiomonas and Rhodanobacter as the key genus closely associated with N2O production, suggesting their potential roles for denitrification. This is the first study to elucidate N2O emission and influential mechanisms in mining-affected rivers using combined isotopic and molecular techniques. The discovery of this study enhances our understanding of the distinctive processes driving N2O production and consumption in highly anthropogenically disturbed aquatic systems, and also provides the foundation for accurate assessment of N2O emissions from mining-affected rivers on regional and global scales.

Modeling coupled nitrification–denitrification in soil with an organic hotspot

Abstract. The emission of nitrous oxide (N2O) from agricultural soils to the atmosphere is a significant contributor to anthropogenic greenhouse gas emissions. The recycling of organic nitrogen (N) in manure and crop residues may result in spatiotemporal variability in N2O production and soil efflux which is difficult to capture by process-based models. We propose a multi-species, reactive transport model to provide detailed insight into the spatiotemporal variability in nitrogen (N) transformations around such N2O hotspots, which consists of kinetic reactions of soil respiration, nitrification, nitrifier denitrification, and denitrification represented by a system of coupled partial differential equations. The model was tested with results from an incubation experiment at two different soil moisture levels (−30 and −100 hPa) and was shown to reproduce the recorded N2O and dinitrogen (N2) emissions and the dynamics of important carbon (C) and N components in soil reasonably well. The simulation indicated that the four different microbial populations developed in closely connected but separate layers, with denitrifying bacteria growing within the manure-dominated zone and nitrifying bacteria in the well-aerated soil outside the manure zone and with time also within the manure layer. The modeled N2O production within the manure zone was greatly enhanced by the combined effect of oxygen deficit, abundant carbon source, and supply of nitrogenous substrates. In the wetter soil treatment with a water potential of −30 hPa, the diffusive flux of nitrate (NO3-) across the manure–soil interface was the main source of NO3- for denitrification in the manure zone, while at a soil water potential of −100 hPa, diffusion became less dominant and overtaken by the co-occurrence of nitrification and denitrification in the manure zone. Scenarios were analyzed where the diffusive transport of dissolved organic carbon or different mineral N species was switched off, and they showed that the simultaneous diffusion of NO3-, ammonium (NH4+), and nitrite (NO2-) was crucial to simulate the dynamics of N transformations and N2O emissions in the model. Without considering solute diffusion in process-based N2O models, the rapid turnover of C and N associated with organic hotspots can not be accounted for, and it may result in the underestimation of N2O emissions from soil after manure application. The model and its parameters allow for new detailed insights into the interactions between transport and microbial transformations associated with N2O emissions in heterogeneous soil environments.

Major challenges in widespread adaptation of aerobic rice system and potential opportunities for future sustainability

Climate change and diminishing resources are the major challenges in sustainable rice production. The aerobic rice system can be a transformational approach to replace conventional rice in the face of climate change and scarcity of resources due to its high input-use efficiency. However, it could not be adapted widely yet due to several challenges like higher weed infestation and low nitrogen (N) use efficiencies. This transformed system can only be adapted at a wider scale through investigation of associated constraints and risks and provision of potential adjustive measures. Aerobic rice crop failure can be reduced if the availability of suitable and stress-resistant cultivars with early improved vigour, and short-duration characteristics are ensured for maintaining the reproductive capability under resource stressed environment. This transformed rice system exhibits limited response to applied N because of increased losses due to higher volatilization and rapid coupled nitrification-denitrification processes under alternate wetting and drying systems. Shifting from the flooded to aerobic rice system bringing changes in crop water demand, organic matter (OM) turnover, N dynamics, weed flora, and greenhouse gases (GHGs) emissions. Along with N-losses, continuous monocropping, higher weed infestation, weedy rice emergence, nitrous oxide (N2O) emissions, nutrient disorders, higher prevalence of pathogens and diseases, increased panicle sterility, and an increase in soilborne pathogens lodging are other major constraints negatively impacting the widespread adaptation of aerobic rice. This article reviews the climate change status, its associated impacts on the sustainability of traditional rice systems, and the rationale for transforming from traditional to aerobic rice systems. Furthermore, it describes the major challenges associated with the aerobic rice system, thereby presenting management strategies ensuring its wider adaptation for sustainable rice production in the face of projected climate change and scarcity of resources.

Denitrification is not Necessarily the Main Source of N2O from Rewetted Fens

Drained agricultural peatlands are being increasingly rewetted for global warming mitigation. This creates novel ecosystems, with unclear effects on nitrogen cycling. Therefore, we aim to understand the impact of rewetting on nitrous oxide (N2O) production and its sources. Soil samples from pairs of sites differing in water regime (drained [D] and rewetted [W]) and peatland type (coastal fen [C], percolation fen [P] and alder forest [A]) in North-Eastern Germany were analyzed for microbial production pathways of N2O using the dual-isotope method with four tracers (H218O, N18O3−, 15NO3−, 15NH4+) in a laboratory incubation experiment. Unexpectedly, the largest N2O fluxes were found for rewetted sites. In four sites, denitrification dominated N2O production (80—90%). Only CW and AD displayed almost equal contributions of N2O from NO3− and NH4+, showing also largest maximum contributions of nitrifier denitrification (44–48%). Nitrification contributed less than 8% in all soils. Less than 20% of N2O was from nitrification-coupled denitrification. Soil samples with high initial water content, requiring drying prior to preincubation, displayed largest emissions, irrespective of peatland type or field water regime. Interestingly, if field conditions were dry and water was added for the preincubation, the contribution of nitrifiers to N2O production was increased, in line with larger concentrations of NO3−. The results confirm the enhancing effect of drainage on N2O fluxes. However, they also indicate a legacy effect of previous conditions on sources of N2O. Overall, short-term changes in water content had strong effects on fluxes, but not sources of N2O.

Antibiotics sulfamethoxazole alter nitrous oxide production and pathways in estuarine sediments: Evidenced by the N15-O18 isotopes tracing

Estuarine antibiotic residues are profoundly impacting microbial nitrogen (N) cycling and associated N2O production, but the response of N2O production pathways to antibiotics remains poorly understood. Here, 15N–18O labeling technique combined with molecular methods were used to investigate the impacts of sulfamethoxazole on the contribution of ammonia oxidation (nitrifier nitrification, nitrifier denitrification, and nitrification-coupled denitrification) and heterotrophic denitrification (HD) to N2O production in estuarine sediments. Results showed that environmental concentration of sulfamethoxazole (4 ng/g) promoted the total N2O production by 17.1% through nitrifier denitrification. Environmentally relevant (40–4000 ng/g) and irrelevant (40,000 ng/g) concentration of sulfamethoxazole drove nitrification denitrification to gradually lose the dominant role in total N2O production and ammonia oxidation-derived N2O, replaced by HD and nitrifier nitrification, while total N2O production were inhibited. Furthermore, when HD dominated the total N2O production, the HD-derived N2O increased by 63.6% with sulfamethoxazole concentration reaching 40,000 ng/g. The mechanistic investigation further showed that nitrifying bacteria were more susceptible to sulfamethoxazole than nitrifying archaea and denitrifiers. The increased expression of nirS gene carried by non-dominant denitrifiers improved the ratio of nirS:nosZ and hence increased HD-derived N2O under high sulfamethoxazole stresses. Overall, our results provide a comprehensive view into how antibiotics regulate N2O production and its pathways in estuarine sediments.

Weak Hypoxia Enhanced Denitrification in a Dissimilatory Nitrate Reduction to Ammonium (DNRA)-Dominated Shallow and Eutrophic Coastal Waterbody, Jinhae Bay, South Korea

The effects of seasonal hypoxia on sediment-water interface nitrogen (N) transformations in Jinhae Bay were examined from 2015 to 2019. The rates of benthic denitrification, anaerobic ammonium oxidation (anammox), dissimilatory nitrate reduction to ammonium (DNRA), nutrient exchange, and sediment oxygen consumption were measured seasonally. The oxygen (O2) and hydrogen sulfide (H2S) depth profiles were measured using microelectrodes. Neither penetration nor consumption of oxygen decreased during hypoxia. Denitrification, anammox, and DNRA ranged from 0 to 0.73, 0.13, and 1.09 mmol N m-2 day-1, respectively. Denitrification, the dominant N removal pathway, increased by 75% while anammox ceased, which led to an overall increase of 55% in the total N2 gas production during hypoxia relative to that during normoxia. Enhanced denitrification is the result of increased coupled nitrification–denitrification due to the intermittent supply of oxygen during bottom water hypoxia (“weak hypoxia”). In the hypoxic period, DNRA decreased by 62%, and the relative contribution of DNRA to the total nitrogen reduction process decreased from 81 to 58%, but it still outperformed denitrification as the main nitrate reduction pathway. Sediments were strong sources of ammonium for the water column, both under normoxia and hypoxia, whereas they were a sink of nitrate from the water column during hypoxia. Bioturbation may be important for maintaining oxygen penetration and consumption in sediments. The dominance of DNRA was mainly due to the relatively high content of sulfide and organic-rich sediments. The repressed macrofaunal activity and increased coupling of nitrification and denitrification during hypoxia may have contributed to enhanced denitrification. Taken together, the overall dominance of DNRA might contribute to the development and maintenance of eutrophication and seasonal hypoxia in this system. However, in contrast to the previous results, denitrification was enhanced during “weak hypoxia,” which might be helpful in alleviating eutrophication.

Benthic exchange along a tropical estuarine salinity gradient during dry season: Biogeochemical and ecological implications

Benthic biogeochemistry and its coupling with pelagic ecology are less understood in tropical than in temperate estuaries, which have significant implications for global carbon and nutrient cycling. In dry season, the Mandovi Estuary of Western India develops a prominent salinity gradient (0–35 psu), and remains nutrient-limited but most productive, which stands in contrast to wet season. To understand the role of benthic exchange in the sustenance of estuarine biogeochemistry and ecosystem, we carried out biogeochemical measurements in the water column and a series of intact-core incubations along the estuarine salinity gradient, during the dry season of 2014. We observed that the lower estuary was nitrogen (N)-limited whereas the middle and upper estuary were phosphorus (P)-limited. The benthic respiration (38.6–82.87 mmol O2 m−2 d−1), organic carbon (Corg) mineralization (34.3–79.8 mmol C m−2 d−1) and nutrient exchange rates considerably varied along the salinity gradient. Variability in benthic respiration was apparently controlled by labile Corg content and faunal abundance in the sediments. Benthic exchange of NH4+ and NOx− were mainly controlled by salinity and NOx− of the water column, respectively. Benthic PO43− exchange was predominantly controlled by sedimentary Fe content and bottom water O2, whereas benthic SiO44− exchange was primarily controlled by sediment-water SiO44− gradient. Abiotic processes accounted for 37-79% of sediment O2 consumption (SOC). Benthic metabolism accounted for 34–60% of total community respiration and mineralized 43–145% equivalent of C fixed through primary production (PP). Of dissolved inorganic nitrogen (DIN) efflux (0.67–2.59 mmol m−2 d−1), NH4+ comprised 91% in the lower estuary but NOx− (NO3− + NO2−) comprised 97% in the upper estuary, and benthic nitrification apparently determined the dominant N species released from the sediments. Coupled nitrification-denitrification apparently caused the loss of 30–69% of NH4+ diffusing from the deeper sediments. The estuarine sediments were a net source of DIN and SiO44−, potentially meeting up to 31% of N and 40% of Si demand of the phytoplankton, but acted as a net PO43− sink owing to high sedimentary Fe and normoxia, and tended to buffer the estuarine water PO43− to 0.05–0.19 μM. Benthic PO43− influx (0.006–0.026 mmol m−2 d−1) apparently caused the P-limiting condition in the middle and upper estuary. Particularly, the role benthic DIN supply was crucial for the sustenance of estuarine PP as the estuary receives some PO43− from the sea and sufficient SiO44− from the river but not sufficient DIN. The sediments potentially immobilized up to 25% and 20% of riverine DIN and PO43− load, respectively. Overall, the study revealed an efficient benthic carbon recycling and a tight benthic-pelagic coupling in the estuary during the dry season.

Nitrogen removal processes coupled with nitrification in coastal sediments off the north East China Sea

Denitrification and anaerobic ammonia oxidation (anammox) play key roles in nitrogen (N) loss, and nitrification can supply substrates of NO2– and NO3– for denitrification and anammox. Coupled nitrification-denitrification/anammox processes are thus crucial for N removal in coastal ecosystems. This study aims to examine the spatial-temporal variations of ambient, coupled, and uncoupled N removal rates in the coastal sediments off the north East China Sea, and to clarify the controlling factors and microbial mechanisms of coupled nitrification-denitrification/anammox. The rates of ambient, coupled, and uncoupled denitrification and anammox in coastal sediments off the north East China Sea were quantified using the continuous-flow experiments combined with 15N isotope pairing technique. The quantitative polymerase chain reaction method was used to determine the abundances of nitrifiers, denitrifiers, and anammox bacteria, with the functional genes of amoA and nirS, and 16S rRNA gene, respectively. Ambient denitrification rates varied between 0.43 and 7.39 μmol N m−2 h−1, and ambient anammox rates ranged from 0.05 to 0.62 μmol N m−2 h−1. Coupled nitrification-denitrification was the dominant N removal pathway. The rates and coupling of N removal processes with nitrification varied distinctly between nearshore and offshore sites, which were driven by diverse environmental factors. Redundancy analysis suggested that nitrate and sulfide were important factors controlling the coupled and uncoupled N removal rates, and nitrate was proved to be the key factor influencing the ratio between coupled and uncoupled N removal via an integrated analysis. Abundances of ammonia oxidizing bacteria (AOB) correlated significantly with coupled denitrification rates and abundances of denitrifiers, suggesting the importance of AOB in coupled nitrification-denitrification. This study investigated the ambient, coupled, and uncoupled denitrification and anammox rates in coastal sediments off the north East China Sea. Nitrate was proved to be the critical factor influencing the ratio between coupled and uncoupled N removal, and AOB may play important role in coupled nitrification-denitrification. These results emphasized that nitrification is crucial for N removal with important implications on N loss in coastal ecosystems.

Biochar amendment mitigated N2O emissions from paddy field during the wheat growing season

Biochar may variably impact nitrogen (N) transformation and N-cycle-related microbial activities. Yet the mechanism of biochar amendment on nitrous oxide (N2O) emissions from agricultural ecosystems remains unclear. Based on a 6-year long-term biochar amendment experiment, we applied a dual isotope (15N–18O) labeling technique with tracing transcriptional genes to differentiate the contribution of nitrifier nitrification (NN), nitrifier denitrification (ND), nitrification-coupled denitrification (NCD) and heterotrophic denitrification (HD) pathway to N2O production. Then the field experiment provided quantitative data on dynamic N2O emissions, soil mineral N and key functional marker gene abundances during the wheat growing season. By using 15N–18O isotope, biochar decreased N2O emission derived from ND (by 45–94%), HD (by 35–46%) and NCD (by 30–64%) compared to the values under N application. Biochar increased the relative contribution of NN to total N2O production as evidenced by the increase in ammonia-oxidizing bacteria, but did not influence the cumulative NN-derived N2O. The field experiment found that the majority of the N2O emissions peaked following fertilization, in parallel with soil NH4+ and nitrite dynamics. Soil N2O emissions during the wheat growing stage were effectively decreased (by 38–48%) by biochar amendment. Based on the correlation analyses and random forest analysis in both microcosm and field experiments, the decrease in nitrite concentration (by 62–65%) and increase in N2O consumption were mainly responsible for net N2O mitigation, as evidenced by the decrease in the ratios of nitrite reductase genes/transcripts (nirS, nirK and fungal nirK) and N2O reductase gene/transcripts (nosZI and nosZII). Based on the extrapolation from microcosm to field, biochar significantly mitigated N2O emissions by weakening the ND processes, since NCD and HD contributed little during the N2O emission “peaks” following urea fertilization. Therefore, emphasis should be put on the ND process and nitrite accumulation during N2O emission peaks and extrapolated to all agroecosystems.

Nitrification and denitrification in recirculating aquaculture systems: the processes and players

AbstractOne of the major challenges in any sustainable aquaculture production systems is the accumulation of nitrogenous waste such as ammonia and its biological nitrification products viz nitrite and nitrate. Considering the bio‐security issues, amelioration of these wastes without water exchange can be accomplished only by way of establishing in situ nitrification and denitrification through biofilters/bioreactors activated with nitrifying/denitrifying bioaugmentors. In such systems, coexistence of aerobic denitrifiers, anaerobic ammonia oxidizers (Anammox) and complete ammonia oxidizers (Comammox) together with the autotrophic nitrifiers enhance the coupled nitrification–denitrification. This promotes total nitrogen removal without external carbon supplements or additional anerobic compartment in the system. Various recirculating aquaculture systems (RAS) comprise diverse nitrifying community in biofilters/bioreactors thereby imparting distinctive nitrogen conversion in the system. Meanwhile, the structure and population dynamics of the nitrifying/denitrifying consortia are influenced by the environmental factors forming the decisive factors of the success of the processes. Accordingly, understanding the complexity of nitrifying/denitrifying community composition turns out to be a requirement to facilitate its improvised performance. In this context, the review addresses different biological nitrogen removal systems, significance of nitrification–denitrification in RAS, genetic diversity of the key players in RAS, methods of analysis of their community structure, current application and prospects of the processes in RAS.

Nitrogen Dynamics of Inundated Sediments in an Emerging Coastal Deltaic Floodplain in Mississippi River Delta Using Isotope Pairing Technique to Test Response to Nitrate Enrichment and Sediment Organic Matter

Nitrate (NO3–) enrichment in rivers over the last four decades has potentially shifted how coastal ecosystems process nitrogen (N). Shifts in N dynamics may be particularly significant in coastal deltaic floodplains when sediments are inundated during river flood stage as this may change the fate of NO3– transported to the coastal ocean. We evaluated the relative importance of denitrification, dissimilative nitrate reduction to ammonium (DNRA), and anaerobic ammonium oxidation (anammox) in intact sediment cores of Wax Lake Delta (WLD) using continuous flow-through system. We manipulated this experimental system with two concentrations of riverine NO3– (lower concentration at 5 μM and ambient concentration at 100 μM) to test how nitrate-enriched waters may modify N cycling in coastal deltaic floodplains. Inundated sediments in a coastal deltaic floodplain removed bio-reactive N as a function of sediment organic matter (SOM) concentrations, indicating how N dynamics vary as a result of deltaic succession. Direct denitrification was the dominant N pathway in inundated sediments in WLD, which was several times greater than coupled nitrification-denitrification, DNRA, and anammox. Gross denitrification (direct denitrification + coupled nitrification-denitrification) rates generally increased with SOM concentrations and ranged from 4 to 12 μmol N m–2 h–1 under lower NO3– enrichment (5 μM) and 68 to 276 μmol N m–2 h–1 under higher NO3– enrichment (100 μM). Most of these changes in N fluxes with SOM and NO3– enrichment resulted from the increase in direct denitrification rates rather than coupled nitrification-denitrification rates. DNRA rates varied from 0 to 65 μmol N m–2 h–1, and the relative significance of DNRA increased as a percentage of total NO3– reduction (%DNRA) with increasing ratio of organic carbon to NO3– concentrations (OC/NO3–). Anammox rates increased among sites as SOM increased, but rates were lower from 0.0 to 38 μmol N m–2 h–1. The dominance of direct denitrification and increased anammox rates with SOM concentrations indicate that dynamic deltaic floodplains can permanently remove bio-reactive N in the proximal zone of river deposition of tidal freshwater wetlands.

Time-resolved characteristics and production pathways of simulated landfilling N2O emission under different oxygen concentrations

Nitrous oxide (N2O), an important greenhouse gas, is emitted from landfill reservoirs, especially in the working face, where nitrification and denitrification occur under different O2 concentrations. In order to explore the effects of O2 concentration on N2O emissions and production pathways, the production of N2O from simulated fresh waste landfilling under 0%, 5%, 10%, and 21% (vol/vol) O2 concentrations were examined, and 15N isotopes were used as tracers to determine the contributions of nitrification (NF), heterotrophic denitrification (HD), and nitrification-coupled denitrification (NCD) to N2O production over a 72-h incubation period. Equal amounts of total nitrogen consumption occurred for all studied O2 concentration and the simulated waste tended to release more N2O under 0% and 21% O2. Heterotrophic denitrification was the main source of N2O release at the studied oxygen concentrations, contributing 90.51%, 69.04%, 80.75%, and 57.51% of N2O under O2 concentrations of 0%, 5%, 10%, and 21%, respectively. Only denitrification was observed in the simulated fresh waste when the oxygen concentration of the bulk atmosphere was 0%. The nitrate reductase (nirS)-encoding denitrifiers in the simulated landfill were also studied and significant differences were observed in the richness and diversity of the denitrifying community at different taxonomic levels. It was determined that optimising the O2 content is a crucial factor in N2O production that may allow greenhouse gas emissions and N turnover during landfill aeration to be minimised.

Denitrification, anammox, and dissimilatory nitrate reduction to ammonium across a mosaic of estuarine benthic habitats

AbstractEstuaries play a key role in moderating the flow of nitrogen (N) to marine ecosystems. However, the magnitude of this N removal can vary dramatically both within and between estuaries due to the benthic habitats present. Here, we compare denitrification, coupled nitrification–denitrification, anammox, and dissimilatory nitrate reduction to ammonium (DNRA) across a mosaic of benthic habitats in the subtropical Noosa River Estuary, Australia. Using 15N tracer techniques and passive pore‐water samplers, we show that coupled nitrification–denitrification was the dominant pathway for N2 production across all habitats, with higher rates in vegetated habitats (10–70 μmol N m−2 h−1) compared to bare sediments (0.9–2 μmol N m−2 h−1). Unusual pore‐water profiles in the macroalgal sediments suggest the presence of sulfur‐driven anoxic nitrification of NH4+ to NO3− and N2. A benthic N budget showed that combined denitrification and coupled nitrification–denitrification accounted approximately 96% of the N2 production, while DNRA accounted for 9% of total NO3− reduction pathways in the Noosa River Estuary. The macroalgae habitat contributed 76% of total N removal via N2 production and 65% of N retention via DNRA, despite accounting for only 25% of the total surface area. We show a strong relationship between seagrass and macroalgae area and N2 production (r2 = 0.8; p < 0.01), and as such, the capacity to mitigate reactive N loads in estuaries may decrease with the large‐scale loss of seagrass and other vegetated habitats.

Deep-sea sponge grounds as nutrient sinks: denitrification is common in boreo-Arctic sponges

Abstract. Sponges are commonly known as general nutrient providers for the marine ecosystem, recycling organic matter into various forms of bioavailable nutrients such as ammonium and nitrate. In this study we challenge this view. We show that nutrient removal through microbial denitrification is a common feature in six cold-water sponge species from boreal and Arctic sponge grounds. Denitrification rates were quantified by incubating sponge tissue sections with 15NO3--amended oxygen-saturated seawater, mimicking conditions in pumping sponges, and de-oxygenated seawater, mimicking non-pumping sponges. It was not possible to detect any rates of anaerobic ammonium oxidation (anammox) using incubations with 15NH4+. Denitrification rates of the different sponge species ranged from below detection to 97 nmol N cm−3 sponge d−1 under oxic conditions, and from 24 to 279 nmol N cm−3 sponge d−1 under anoxic conditions. A positive relationship between the highest potential rates of denitrification (in the absence of oxygen) and the species-specific abundances of nirS and nirK genes encoding nitrite reductase, a key enzyme for denitrification, suggests that the denitrifying community in these sponge species is active and prepared for denitrification. The lack of a lag phase in the linear accumulation of the 15N-labelled N2 gas in any of our tissue incubations is another indicator for an active community of denitrifiers in the investigated sponge species. Low rates for coupled nitrification–denitrification indicate that also under oxic conditions, the nitrate used to fuel denitrification rates was derived rather from the ambient seawater than from sponge nitrification. The lack of nifH genes encoding nitrogenase, the key enzyme for nitrogen fixation, shows that the nitrogen cycle is not closed in the sponge grounds. The denitrified nitrogen, no matter its origin, is then no longer available as a nutrient for the marine ecosystem. These results suggest a high potential denitrification capacity of deep-sea sponge grounds based on typical sponge biomass on boreal and Arctic sponge grounds, with areal denitrification rates of 0.6 mmol N m−2 d−1 assuming non-pumping sponges and still 0.3 mmol N m−2 d−1 assuming pumping sponges. This is well within the range of denitrification rates of continental shelf sediments. Anthropogenic impact and global change processes affecting the sponge redox state may thus lead to deep-sea sponge grounds changing their role in marine ecosystem from being mainly nutrient sources to becoming mainly nutrient sinks.

Effects of silver nanoparticles on coupled nitrification–denitrification in suspended sediments

The effects of varying concentrations of Ag NPs on coupled nitrification and denitrification (CND) in two suspended sediments (SPSs) sizes were investigated using isotopic tracer method. In general, 0.5 and 5 mg/L Ag NPs had less effect on CND, while 2 and 10 mg/L Ag NPs exhibited the improvement and inhibition effect, respectively. The CND improvement by 2 mg/L NPs was mainly due to the enhanced nitrifying and denitrifying enzyme activity. However, 10 mg/L Ag NPs inhibited NH4+ oxidation by directly reducing the AMO activity and AOB abundance. The inhibition on NAR and NIR activity and their encoding narG and nirK gene abundance further inhibited NO3− and NO2− reduction, leading to a dramatic decrease in the 15N–N2 production. The above inhibition effects were attributed to the nano-effects of Ag NPs, which led to the excessive ROS amount and the decreased T-AOC level in microbial systems. But the connection between nitrification and denitrification was not broken after Ag NPs exposure. Moreover, the results indicated that N-cycling in clay and silt-type SPS systems could be more sensitive than sand-type SPS systems to NP exposure. The findings provide a basis for evaluating the environmental risks of Ag NPs in water–sediment systems.

The Effect of Chironomid Larvae on Nitrogen Cycling and Microbial Communities in Soft Sediments

The combination of biogeochemical methods and molecular techniques has the potential to uncover the black-box of the nitrogen (N) cycle in bioturbated sediments. Advanced biogeochemical methods allow the quantification of the process rates of different microbial processes, whereas molecular tools allow the analysis of microbial diversity (16S rRNA metabarcoding) and activity (marker genes and transcripts) in biogeochemical hot-spots such as the burrow wall or macrofauna guts. By combining biogeochemical and molecular techniques, we analyzed the role of tube-dwelling Chironomus plumosus (Insecta, Diptera) larvae on nitrification and nitrate reduction processes in a laboratory experiment with reconstructed sediments. We hypothesized that chironomid larvae stimulate these processes and host bacteria actively involved in N-cycling. Our results suggest that chironomid larvae significantly enhance the recycling of ammonium (80.5 ± 48.7 µmol m−2 h−1) and the production of dinitrogen (420.2 ± 21.4 µmol m−2 h−1) via coupled nitrification–denitrification and the consumption of water column nitrates. Besides creating oxygen microniches in ammonium-rich subsurface sediments via burrow digging and ventilation, chironomid larvae serve as hot-spots of microbial communities involved in N-cycling. The quantification of functional genes showed a significantly higher potential for microbial denitrification and nitrate ammonification in larvae as compared to surrounding sediments. Future studies may further scrutinize N transformation rates associated with intimate macrofaunal–bacteria associations.

Annual dynamics of soil gross nitrogen turnover and nitrous oxide emissions in an alpine shrub meadow

Soil nitrogen (N) transformations play a vital role in maintaining grassland productivity and influence nitrous oxide (N2O) emissions. To evaluate annual dynamics of soil gross N turnover and its effects on N2O emissions in typical grasslands, we conducted year-round measurements of soil gross N turnover rates, inorganic N pool sizes and N2O fluxes in an alpine shrub meadow of the Qinghai–Tibet Plateau. Gross ammonium (NH4+) and nitrate (NO3−) immobilization rates were calculated by the “isotope dilution method” (i.e., based on the consumption rates) and the “reformed difference method” (i.e., based on the differences between gross and net turnover rates). The “reformed difference method” avoids additional measurements of net N turnover rates in unlabeled soils compared to the traditional “difference method”, and provides more reliable estimates of NH4+ immobilization in soils with low ambient NH4+ pool sizes than the “isotope dilution method”. The annual gross rates of mineralization, nitrification, NH4+ and NO3− immobilization amounted to 606 ± 43, 236 ± 27, 389 ± 24 and 82 ± 19 mg N kg−1 dry soil yr−1, respectively, in a topsoil of 10 cm. The soil gross N turnover rates during freezing and freeze–thaw periods contributed considerably to the annual totals (22–44%). Thus, the freezing and freeze–thaw periods were not dormant seasons for microbial N transformations, even under a frigid alpine climate. The annual N2O emission was 0.20 ± 0.03 kg N ha−1 yr−1, 35% of which originated from a short-lived emission pulse during the freeze–thaw period (52 d). The promotion of gross mineralization increased soil NH4+ pool sizes, whereas the increase in gross nitrification did not enhance soil NO3− pool sizes during the freeze–thaw period. Coupled nitrification–denitrification dominated NO3− consumption and freeze–thaw related N2O production and hence, the low NO3− pool sizes were not indicative of the potential of pulsed N2O emissions. Year-round measurements with intensive observations during both the non-freezing and freeze–thaw periods are indispensable to fully understand soil N cycling and its response to climate change in alpine ecosystems.