N2O Production Pathways Research Articles

Forecasting nitrous oxide emissions from a full-scale wastewater treatment plant using LSTM-based deep learning models

Nitrous oxide (N2O) emissions from wastewater treatment plants (WWTPs) exhibit significant seasonal variability, making accurate predictions with conventional biokinetic models difficult due to complex and poorly understood biochemical processes. This study addresses these challenges by exploring data-driven alternatives, using long short-term memory (LSTM) based encoder-decoder models as basis. The models were developed for future integration into a model predictive control framework, aiming to reduce N2O emissions by forecasting these over varying prediction horizons. The models were trained on 12 months and tested on 3 months of data from a full-scale WWTP in Amsterdam West, the Netherlands. The dataset encompasses seasonal peaks in N2O emissions typical for winter and spring months. The best performing model, featuring a 256-256 LSTM architecture, achieved the highest accuracy with test R2 values up to 0.98 across prediction horizons spanning 0.5 to 6.0 hours ahead. Feature importance analysis identified past N2O emissions, influent flowrate, NH4+, NOx, and dissolved oxygen (DO) in the aerobic tank as most significant inputs. The observed decreasing influence of historical N2O emissions over extended prediction horizons highlights the importance and significance of process variables for the model's performance. The model's ability to accurately forecast short-term N2O emissions and capture immediate trends highlights its potential for operational use in controlling emissions in WWTPs. Further research incorporating diverse datasets and biochemical process inputs related to microbial activities in the N2O production pathways could improve the model's accuracy for longer forecasting horizons. These findings advocate for hybridising deep learning models with biokinetic and mechanistic insights to enhance prediction accuracy and interpretability.

Unprecedented N2O production by nitrate-ammonifying Geobacteraceae with distinctive N2O isotopocule signatures.

Dissimilatory nitrate reduction to ammonium (DNRA), driven by nitrate-ammonifying bacteria, is an increasingly appreciated nitrogen-cycling pathway in terrestrial ecosystems. This process reportedly generates nitrous oxide (N2O), a strong greenhouse gas with ozone-depleting effects. However, it remains poorly understood how N2O is produced by environmental nitrate-ammonifiers and how to identify DNRA-derived N2O. In this study, we characterize two novel enzymatic pathways responsible for N2O production in Geobacteraceae strains, which are predominant nitrate-ammonifying bacteria in paddy soils. The first pathway involves a membrane-bound nitrate reductase (Nar) and a hybrid cluster protein complex (Hcp-Hcr) that catalyzes the conversion of NO2- to NO and subsequently to N2O. The second pathway is observed in Nar-deficient bacteria, where the nitrite reductase (NrfA) generates NO, which is then reduced to N2O by Hcp-Hcr. These enzyme combinations are prevalent across the domain Bacteria. Moreover, we observe distinctive isotopocule signatures of DNRA-derived N2O from other established N2O production pathways, especially through the highest 15N-site preference (SP) values (43.0‰-49.9‰) reported so far, indicating a robust means for N2O source partitioning. Our findings demonstrate two novel N2O production pathways in DNRA that can be isotopically distinguished from other pathways.IMPORTANCEStimulation of DNRA is a promising strategy to improve fertilizer efficiency and reduce N2O emission in agriculture soils. This process converts water-leachable NO3- and NO2- into soil-adsorbable NH4+, thereby alleviating nitrogen loss and N2O emission resulting from denitrification. However, several studies have noted that DNRA can also be a source of N2O, contributing to global warming. This contribution is often masked by other N2O generation processes, leading to a limited understanding of DNRA as an N2O source. Our study reveals two widespread yet overlooked N2O production pathways in Geobacteraceae, the predominant DNRA bacteria in paddy soils, along with their distinctive isotopocule signatures. These findings offer novel insights into the role of the DNRA bacteria in N2O production and underscore the significance of N2O isotopocule signatures in microbial N2O source tracking.

Biogas Residue Carbonization Rather Than Biogas Residue Promoted the Yield of Pakchoi and Reduced the N2O Production Potential in Horticultural Soil

ABSTRACTBiogas residue (BR) and biogas residue‐derived biochar (BRC) are widely used as substitute for inorganic Nitrogen (N) fertilizers in vegetable production. Yet, their comparative research on vegetable production and N2O production was still lacking. Here, the pot experiment of pakchoi (Brassica chinensis) with a gradient of BR or BRC application rates (0, 30%, 60%, 100% w/w) was carried out to simulate different N fertilizer substitution rates. The results showed that the pakchoi yield had no difference between BR or BRC and control treatments; BRC had more advantages than BR in maintaining or increasing the pakchoi yield. BR and BRC could all reduce N2O production potential in vegetable soils. But BRC had a stronger ability to inhibit denitrification while BR had a stronger ability to inhibit nitrification compared with each other. The results showed that BR and BRC had different regulatory pathways for pakchoi yield and N2O production. BR regulated the pakchoi yield majorly through nitrification, but BRC majorly through denitrification. It suggested that BR and BRC could partially or completely replace inorganic fertilizers without reducing pakchoi yield. BRC combined with chemical fertilizers was a higher intelligence strategy in vegetable systems to improve pakchoi yield and N2O production compared with BR. It provided a theoretical basis for the application of BR and BRC to nutrient cycling and microbial processes in the soil‐vegetable system.

Mapping N2O production pathways in estuarine and coastal sediments through coupled 15N tracing and N2O isotopocules analysis

Estuarine and coastal zones are important sources of atmospheric nitrous oxide (N2O). Previous studies have explored the overarching roles of nitrification and denitrification in N2O production from these critical zones, yet the specific productions of various pathways within these processes remain elusive. Using dual 15N tracers (15NH4+ and 15NO3-) coupled with N2O isotopocules analysis, we quantified the contributions of multiple nitrification (both autotrophic and heterotrophic) and denitrification (bacterial, fungal, and chemical) pathways to N2O production in estuarine and coastal sediments. Furthermore, we leveraged microecology theory to dissect the biogeochemical mechanisms that govern the spatiotemporal dynamics of these pathways. Results showed that denitrification was the dominant process, accounting for over 70 % of N2O production. The remainder was attributed to autotrophic nitrification (5.80–23.92 %) and heterotrophic nitrification (1.07–7.10 %). Isotopocules analyses further showed that fungal denitrification was the primary source at freshwater sites (40.47–49.99 %), whereas bacterial denitrification prevailed at high-salinity sites (51.37–52.55 %). Moreover, these processes displayed contrasting spatial variability along the estuarine salinity gradient. Mechanistic investigation informed by microecology theory demonstrated that distinct assembly processes governed bacterial and fungal communities across this gradient, resulting in divergent ecological adaptation strategies of bacterial and fungal denitrifiers and, consequently, the contrasting contribution patterns of bacterial and fungal denitrification. Collectively, our work presents a comprehensive and refined picture of N2O sources and dynamics, offering essential insights for improving predictive models and formulating effective mitigation strategies targeting N2O emissions from estuarine and coastal zones.

Nitrous oxide production and consumption by marine ammonia-oxidizing archaea under oxygen depletion.

Ammonia-oxidizing archaea (AOA) are key players in the nitrogen cycle and among the most abundant microorganisms in the ocean, thriving even in oxygen-depleted ecosystems. AOA produce the greenhouse gas nitrous oxide (N2O) as a byproduct of ammonia oxidation. Additionally, the recent discovery of a nitric oxide dismutation pathway in the AOA isolate Nitrosopumilus maritimus points toward other N2O production and consumption pathways in AOA. AOA that perform NO dismutation when exposed to oxygen depletion, produce oxygen and dinitrogen as final products. Based on the transient accumulation of N2O coupled with oxygen accumulation, N2O has been proposed as an intermediate in this novel archaeal pathway. In this study, we spiked N2O to oxygen-depleted incubations with pure cultures of two marine AOA isolates that were performing NO dismutation. By using combinations of N compounds with different isotopic signatures (15NO2 - pool +44N2O spike and 14NO2 - pool +46N2O spike), we evaluated the N2O spike effects on the production of oxygen and the isotopic signature of N2 and N2O. The experiments confirmed that N2O is an intermediate in NO dismutation by AOA, distinguishing it from similar pathways in other microbial clades. Furthermore, we showed that AOA rapidly reduce high concentrations of spiked N2O to N2. These findings advance our understanding of microbial N2O production and consumption in oxygen-depleted settings and highlight AOA as potentially important key players in N2O turnover.

Selective increase of antibiotic-resistant denitrifiers drives N2O production in ciprofloxacin-contaminated soils

Agricultural systems significantly contribute to global N2O emissions, which is intensified by excessive fertilization and antibiotic residues, attracting global concerns. However, the dynamics and pathways of antibiotics-induced soil N2O production coupled with microbial metabolism remain controversial. Here, we explored the pathways of N2O production in agricultural soils exposed to ciprofloxacin (CIP), and revealed the underlying mechanisms of CIP degradation and the associated microbial metabolisms using 15N-isotope labeling and molecular techniques. CIP exposure significantly increases the total soil N2O production rate. This is attributed to an unexpected shift from heterotrophic and autotrophic nitrification to denitrification and an increased abundance of denitrifiers Methylobacillus members under CIP exposure. The most striking strain M. flagellatus KT is further discovered to harbor N2O-producing genes but lacks a N2O-reducing gene, thereby stimulating denitrification-based N2O production. Moreover, this denitrifying strain is probably capable of utilizing the byproducts of CIP as carbon sources, evidenced by genes associated with CIP resistance and degradation. Molecular docking further shows that CIP is well ordered in the catalytic active site of CotA laccase, thus affirming the potential for this strain to degrade CIP. These findings advance the mechanistic insights into N2O production within terrestrial ecosystems coupled with the organic contaminants degradation.

Aquacultural source of nitrous oxide revealed by nitrogen isotopes

The rapid expansion of coastal aquaculture has led to an increase in the coverage of aquaculture ponds, where intense feed-derived nitrogen is causing significant emissions of nitrous oxide (N2O). Multiple N2O production pathways and the relative importance of water column vs. sedimentary production in aquaculture ponds remain uncertain. Clarifying these pathways is vital for sustainable aquaculture development. Using 15N-labeled dissolved inorganic nitrogen, the pathways and rates of N2O production in subtropical aquaculture ponds located in south China, cultivating whiteleg shrimp, Japanese seabass, and giant river prawn, were successfully characterized. Total N2O production rates ranged from 6 to 70 µmol-N m−2 d−1, with the shrimp pond exhibiting the highest total N2O production rates, followed by ponds for seabass and prawn. These differences are primarily due to varying feed amounts causing differences in dissolved nutrients in water column and sediment. Particularly, nutrient and organic matter accumulation at the surface sediment stimulated N2O production. The oxygenated sediment on a centimeter scale could produce substantially more N2O compared to the water column above on a meter scale. Partial denitrification, i.e., nitrate and nitrite reduction to N2O, was more important (> 60 %) for N2O production in aquaculture ponds. The availability of nitrite is likely a major factor driving partial denitrification for both sedimentary and water column N2O production.

Enhancement of nitrogen removal in constructed wetlands through boron mud and elemental sulfur addition: Regulation of sulfur and oxygen cycling

Constructed wetlands (CWs) utilizing elemental sulfur (S0) as an electron donor are effective for nitrate (NO3–-N) removal from low C/N wastewater. However, the limited bioavailability of chemical S0 (chem-S0) due to low water solubility and inadequate mass transfer impedes complete denitrification. In this study, boron mud waste and S0 (S0&BM) were processed and combined to convert chem-S0 to biological S0 (bio-S0) in CWs. The employment of S0&BM improved NO3–-N and total nitrogen (TN) removal rates by 15.36 % and 10.55 % compared to S0 alone. The 15N isotope tracing experiment unveiled the pathways of N2O production and demonstrated that S0&BM facilitated full denitrification, resulting in a higher percentage of nitrogen being degraded in a non-polluting form (54.70 %). Releasing trace amounts of boron and magnesium in S0&BM mitigated the excessive acidification caused by the sulfur autotrophic process and promoted biofilm formation, thereby pronouncing vertical oxygen fluctuations. Meanwhile, oxygen vacancies generated in the S0&BM promoted the closed-loop sulfur cycle, starting from the oxidation of chem-S0 to sulfate (SO42-), the important process of SO42--reducing bacteria using partial organic compounds as electron donors to reduce SO42- to sulfide (S2-) was realized, and finally the S2- was oxidized to bio-S0 with better properties. S0&BM enriched functional microbial communities in the biofilm related to nitrogen and sulfur transformation. Functional gene analysis showed an increase in the narG, nirS, norB, and nosZ genes supporting denitrification. The dsrA and sqr genes responsible for the sulfur cycle exhibited the highest abundance. This realization allowed for the utilization of waste materials for treatment, enhancing the energy efficiency of carbon reduction and pollution reduction in CWs.

Nitrous oxide (N2O) emissions at the air-water-sediment interfaces of cascade reservoirs in the Yunnan-Guizhou Plateau: Spatial patterns and environmental controls

The construction of cascade reservoirs can interfere with the natural hydrologic cycles of basins, causing negative environmental effects such as altering the emission patterns of the Nitrous oxide (N2O), a potent greenhouse gas. To elucidate the impact of cascade reservoirs construction on river N2O emissions, we utilized the thin boundary model and the incubation experiments to estimate the N2O fluxes at the air-water interface and at the water-sediment interface of cascade reservoirs on the Yunnan-Guizhou Plateau, respectively. Additionally, we explored the influence of various factors, with particular emphasis on damming, on N2O emissions and production. Moreover, we identified the main pathways of N2O production and proposed management strategies to mitigate N2O emissions from cascade reservoirs. The findings revealed that N2O fluxes at the air-water interface and the water-sediment interface were 4.73 ± 1.32 μmol m−2 · d−1 and 15.56 ± 1.98 μmol m−2 · d−1, respectively. Influenced by temperature, dissolved oxygen (DO), resource substances (active nitrogen substrates and dissolved organic carbon (DOC)) and reservoir properties (scale, hydraulic retention time (HRT), reservoir age, etc.), the N2O concentration and flux exhibited notable spatial heterogeneity, gradually increasing downstream. Temperature has a significant direct impact on N2O flux, as well as indirect effects through DO and resource chemicals. Furthermore, the correlation between dissolved oxygen utilization rate (AOU) and net N2O flux (ΔN2O) indicated that N2O emissions at the water-air interface were primarily attributable to nitrification, whereas those at the water-sediment interface were predominantly driven by denitrification. These findings not only enhance our comprehension of N2O emissions at various interfaces of cascade reservoirs but also offer theoretical backing for the formulation of management strategies aimed at efficiently mitigating N2O emissions from continuously dammed rivers.

Dissolved Nitrous Oxide and Hydroxylamine in the Northern South China Sea in Summer

AbstractNitrous oxide (N2O) is a potent greenhouse gas and a strong ozone‐depleting substance. Ocean is an important natural source of atmospheric N2O, which has attracted increasing attention recently as to understand the biogeochemical cycles of nitrogen in marine systems. Nitrification is one of the major N2O production pathways during which N2O is produced as a side product and hydroxylamine (NH2OH) is produced as an obligate intermediate. However, few studies have reported the NH2OH variability in natural seawater and its relationship between N2O remains ambiguous. In summer 2022, spatial distribution and influencing factors of N2O and NH2OH in the northern South China Sea (SCS) were investigated. Dissolved N2O varied from 5.79 to 34.80 nmol L−1 in water column and generally increased with water depth above 800 m. N2O was correlated with dissolved oxygen and nitrate, inferring that N2O was mainly produced via nitrification. NH2OH varied from below detection limit to 3.91 nmol L−1. The adoption of multiple standard additions can largely reduce the bias in NH2OH measurement. Structural equation modeling showed that NH2OH has a potential effect favoring excessive N2O. Surface N2O saturation ranged between 102% and 278%, indicating that the northern SCS was a net source of atmospheric N2O in summer.

Long-neglected contribution of nitrification to N2O emissions in the Yellow River

Rivers play a significant role in the global nitrous oxide (N2O) budget. However, the microbial sources and sinks of N2O in river systems are not well understood or quantified, resulting in the prolonged neglect of nitrification. This study investigated the isotopic signatures of N2O, thereby quantifying the microbial source of N2O production and the degree of N2O reduction in the Yellow River. Although denitrification has long been considered to be the dominant pathway of N2O production in rivers, our findings indicated that denitrification only accounted for 18.3% (8.2%–43.0%) of the total contribution to N2O production in the Yellow River, with 50.2%–80.2% being concurrently reduced. The denitrification contribution to N2O production (R2 = 0.44, p < 0.01) and N2O reduction degree (R2 = 0.70, p < 0.01) were positively related to the dissolved organic carbon (DOC) content. Similar to urban rivers and eutrophic lakes, denitrification was the primary process responsible for N2O production (43.0%) in certain reaches with high organic content (DOC = 5.29 mg/L). Nevertheless, the denitrification activity was generally constrained by the availability of electron donors (average DOC = 2.51 mg/L) throughout the Yellow River basin. Consequently, nitrification emerged as the primary contributor in the well-oxygenated Yellow River. Additionally, our findings further distinguished the respective contribution of ammonia-oxidizing bacteria (AOB) and archaea (AOA) to N2O emissions. Although AOB dominated the N2O production in the Yellow River, the AOA specie abundance (AOA/(AOA + AOB)) contributed up to 32.6%, which resulted in 25.6% of the total nitrifier-produced N2O, suggesting a significant occurrence of AOA in the oligotrophic Yellow River. Overall, this study provided a non-invasive approach for quantifying the microbial sources and sinks to N2O emissions, and demonstrated the substantial role of nitrification in the large oligotrophic rivers.

Effects of soil water content at freezing, thaw temperature, and snowmelt infiltration on N2O emissions and denitrifier gene and transcript abundance during a single freeze-thaw event

Climate change-related warming and increased precipitation may alter winter snow cover and thawing events, and therefore, may carry significant consequences for nitrous oxide (N2O) production pathways such as denitrification, and the abundance and expression of denitrifying microorganisms. We used a soil microcosm study to investigate the combined effect of soil thaw temperature, initial water filled pore space (WFPS) prior to soil freezing, and snowmelt infiltration simulated by the addition of water on N2O emission and denitrification rates, soil respiration rate, and the abundance and transcription of denitrifying (nirK, nirS, and nosZ) bacteria during a single freeze-thaw event. Soil respiration rate was primarily controlled by an increase in soil thaw temperature, whereas soil N2O emission and denitrification rates were generally greater in soils with a higher initial WFPS and soil thaw temperature. In contrast, snowmelt infiltration generally had a negligible effect on these rates, which may be related to pre-existing soil conditions that were already conducive to denitrification. Unexpectedly, the nosZ transcript/nosZ gene abundance ratio was lower in soils thawed at 8.0 °C compared to 1.5 °C; however, this may have resulted in a lower N2O reduction, thus explaining the greater levels of N2O emitted from soils thawed at 8.0 °C. Overall, this study demonstrated that increased N2O production during a single freeze-thaw event was primarily linked to antecedent conditions of high initial WFPS, soil thaw temperature, and a synergistic interplay between these two environmental parameters, and provides evidence that an increase in annual temperature and precipitation, along with the timing of precipitation, may further stimulate N2O production pathways.

Acidification Offset Warming-Induced Increase in N2O Production in Estuarine and Coastal Sediments.

Global warming and acidification, induced by a substantial increase in anthropogenic CO2 emissions, are expected to have profound impacts on biogeochemical cycles. However, underlying mechanisms of nitrous oxide (N2O) production in estuarine and coastal sediments remain rarely constrained under warming and acidification. Here, the responses of sediment N2O production pathways to warming and acidification were examined using a series of anoxic incubation experiments. Denitrification and N2O production were largely stimulated by the warming, while N2O production decreased under the acidification as well as the denitrification rate and electron transfer efficiency. Compared to warming alone, the combination of warming and acidification decreased N2O production by 26 ± 4%, which was mainly attributed to the decline of the N2O yield by fungal denitrification. Fungal denitrification was mainly responsible for N2O production under the warming condition, while bacterial denitrification predominated N2O production under the acidification condition. The reduced site preference of N2O under acidification reflects that the dominant pathways of N2O production were likely shifted from fungal to bacterial denitrification. In addition, acidification decreased the diversity and abundance of nirS-type denitrifiers, which were the keystone taxa mediating the low N2O production. Collectively, acidification can decrease sediment N2O yield through shifting the responsible production pathways, partly counteracting the warming-induced increase in N2O emissions, further reducing the positive climate warming feedback loop.

Elevated atmospheric CO2 promotes the contribution of autotrophic nitrification to N2O emissions in a typical summer maize field

Elevated atmospheric carbon dioxide (CO2) concentration generally stimulates nitrous oxide (N2O) emissions from upland soils, leading to a promoted feedback on climate change. However, the underlying mechanisms on how elevated CO2 (eCO2) stimulates N2O emissions from nitrogen (N)-fertilized upland soils remain poorly known. Here, we investigated the effects of eCO2 on soil N2O production pathways and associated gross N transformation rates, as well as on nitrifying and denitrifying microbes. Based on a 13 years of free-air CO2 enrichment (FACE) platform, we found that N2O emissions were significantly increased by 46.7% under eCO2 in a typical summer-maize field. The 15N tracing experiment showed that autotrophic nitrification rate was significantly increased by 11.6% under eCO2, coincided with the significantly higher abundance and shift composition of ammonia-oxidizing bacteria (AOB) under eCO2. Also, eCO2 enhanced gross ammonium (NH4+) immobilization rate but reduced gross nitrate (NO3−) immobilization rate. Autotrophic nitrification accounted for approximately 78% of N2O emissions, followed by heterotrophic nitrification (about 19%) and denitrification (about 3%). It was found that eCO2 increased carbon (C) deposition in soil, which could affect the availability of soil labile organic C and N and alter the composition and activity of nitrifiers. These changes promoted the N2O production derived from autotrophic nitrification, resulting in higher N2O emissions under eCO2. We could deduce that nitrifier denitrification contributed to the increased N2O emissions greatly under eCO2, derived by strongly stimulated ammonia oxidation and microbial respiration (i.e. higher soil CO2 emissions) caused suboxic conditions (i.e. lower soil oxygen (O2) concentration), along with higher abundance ratios of (nirK+nirS)/nosZ. Our results suggest that controlling autotrophic nitrification appropriately is crucial for mitigating N2O emissions from upland soils under rising atmospheric CO2 concentration.

How does biochar influence soil nitrification and nitrification-induced N2O emissions?

Nitrification is a major pathway of N2O production, especially in aerobic soils. The amendment of soils with biochar has been suggested as a promising solution to regulate soil N cycle and reduce N2O emissions. However, there is a lack of comprehensive and quantitative understanding of biochar impacts on soil nitrification and nitrification-induced N2O emissions. In this study, a meta-analysis was conducted using data compiled across 95 peer-reviewed studies. Results showed that biochar in general significantly increased soil nitrification rate by 56 %, with overall no significant effect on nitrification-induced N2O emissions, suggesting that biochar likely restricted the fraction of nitrified N emitted as N2O emissions. The abundance of ammonia-oxidizing bacteria (AOB) was significantly increased by 37 % following biochar addition, but that of ammonia-oxidizing archaea (AOA) did not change significantly, indicating that the impact of biochar on AOB rather than AOA may play an important role in soil nitrification. The impacts of biochar on soil nitrification processes were heterogeneous depending on soil properties. Biochar increased soil nitrification rate and AOB abundance to a larger extent in poorly pH-buffered soils such as those with acidic pH (<5), low organic carbon (<10 g kg−1), or poor texture (rich in either sand or clay), which may be attributed to the liming and structural effects of biochar that regulate soil pH and water-air status. The overall no significant effect of biochar on nitrification-induced N2O emissions was due to a positive effect in acidic soils, a negative effect in alkaline soils, and little effect in neutral soils. This study provides a comprehensive insight into how different factors mediate the response of soil nitrification processes to biochar amendment, which contributes to a new understanding of biochar function in regulating soil N2O emissions, and can assist in designing biochar projects that would benefit soil N cycle while minimizing undesirable side effects.

Unravelling spatiotemporal N2O dynamics in an urbanized estuary system using natural abundance isotopes

Estuaries are strong sources of N2O to the atmosphere; yet we still lack insights into the impact of their biogeochemical dynamics on the emissions of this powerful greenhouse gas. Here, we investigated the spatiotemporal dynamics of the N cycle in an estuary with a focus on the emission mechanisms and pathways of N2O. By coupling N2O isotopocule analysis and substrate NO3− isotope analysis, we found that nutrient availability, oxygen level, salinity gradient and temperature variation were major drivers of the N2O emissions from the Scheldt Estuary. In winter, lower temperature and higher O2 concentration diminished denitrification rates and reduction of N2O to N2, while both were enhanced in warmer summer, causing higher fraction of reduced N2O. As a result, we found comparable N2O fluxes and dissolved concentrations between the two seasons. Decrease in salinity level and increase in NO3− concentration accelerated N2O production when moving upstream of the estuary where more urbanization and higher NO3− from wastewater discharges were found. However, these drivers had no significant effect on the fraction of N2O derived by either denitrification or nitrification and/or fungal denitrification since the fractional proportion of these pathways showed no spatiotemporal variations, remaining around 89 % and 11 %, respectively. These findings challenge the conventional notion that N2O fluxes are generally higher in summer because of higher denitrification rates while confirming that denitrification is the most important pathway of N2O production in the estuaries. Furthermore, our study highlight the importance of combining various isotope analyses to gain in-depth understanding about N2O emission pathways and N cycling in dynamic systems like estuaries.

Microbial mechanism of biochar addition to reduce N2O emissions from soilless substrate systems

The soilless peat-based substrate partially solves the global soil problem in greenhouse vegetable production. However, it still produces serious N2O emissions due to the application of nutrient solutions. The pyrolysis biochar is regarded as an effective measure to reduce soil N2O emissions. However, the effect and mechanism of biochar on N2O emissions from the soilless substrate remain unknown. Therefore, this study set up six treatments by adjusting the ratio of biochar addition of peat-based substrate: 0% (0BC), 2% (2BC), 4% (4BC), 6% (6BC), 8% (8BC) and 10% (10BC) (v/v). The results showed that compared to the control treatment, N2O emissions reduced by 81%, 71%, 51%, 61%, and 75% in the 2BC, 4BC, 6BC, 8BC and 10BC treatments, respectively. In addition, lettuce yield increased by 10% and 7% in the 2BC and 4BC treatments and decreased by 0.5%, 4% and 6% in the 6BC, 8BC and 10BC treatments, respectively. Combining stable isotope technology, qPCR analysis and high-throughput sequencing, five microbial pathways of N2O production, including bacterial and archaea nitrification (BN and AN), denitrification performed by fungi, denitrifier bacteria and nitrifier bacteria (FD, DD and ND), were roughly distinguished. In addition, the extent of N2O reduction was obtained by δ18O vs.δ15NSP map. For all treatments, overall, the DD process (over 50%) was the main process of N2O production and reduction, while ND and AN processes were almost negligible (less 5%). In detail, the decrease of N2O emissions was caused by decreasing the contribution of FD in the 6BC, 8BC and 10BC treatments and reducing the contribution of BN in the 0BC and 2BC treatments. In addition, biochar addition increased the extent of N2O reduction to N2. In summary, the 2% biochar addition presented the greatest extent of N2O reduction to N2 (83%) and the lowest N2O emissions as well as the highest lettuce yields and nitrogen utilization efficiency. Therefore, 2% biochar is deemed the most optimal addition to the peat-based substrate.

Effect of Streptomyces JD211 application on soil physicochemical properties and N2O emission characteristics of rice rhizosphere

Biocontrol agent, as a pollution-free and sustainable plant disease control method, can inhibit the spread of soil-borne diseases and promote the growth of crops. However, there are few studies on the effect of biocontrol agent on N2O emission in rice soil. In this study, after the application of the biocontrol agent Streptomyces JD211, N2O emission from rice soil were measured, and the relationship between the agent and soil N2O emissions were studied in soil chemistry and molecular biology. The results showed that the application of Streptomyces JD211 can significantly reduce the rate of N2O emission from rice soil. The NH4+-N and NO3−-N contents in rice soil decreased in a short period of time after the application of Streptomyces JD211, while the mineral N content in the soil remained stable with rice growth. 16S rRNA gene sequencing and metagenomic sequencing revealed Streptomyces JD211 application mainly increased the relative abundance of Burkholderia and Streptomyces in the soil microbial community, reduced the relative abundance of hao, norB, norC genes, and increased the relative abundance of nosZ and hcp genes. Streptomyces JD211 application promoted N2O transformation and weakened N2O production pathways, which ultimately reduced N2O emissions from rice soils. This study provided new insight of biocontrol agents to regulate soil N2O emissions, which is of great significance for the development and application of biocontrol bacteria and farmland environmental protection.

Modelling greenhouse gas emissions from biological wastewater treatment by GPS-X: The full-scale case study of Corleone (Italy)

Greenhouse gas (GHG) emissions from wastewater treatment plants (WWTPs) can affect climate change and must be measured and reduced. Mathematical modelling is an attractive solution to get a tool for GHG mitigation. However, although many efforts have been made to create reliable tools that can simulate “sustainable” full-scale WWTP operation, these studies are not considered complete enough to include GHG emissions and energy consumption of biological processes under long-term dynamic conditions. In this study, activated sludge model no. 1 (ASM1) was modified to model nitrous oxide (N2O) emissions with a plant-wide modelling approach. The model is novel compared to the state of the art since it includes three steps denitrification, all N2O production pathways and its stripping in an ASM1. The model has been calibrated and validated through long-term water quality and short-term N2O emissions data collected from Corleone (Italy) WWTP. Different dissolved oxygen (DO) concentrations and return sludge (RAS) ratios were tested with dynamic simulations to optimise the full-scale WWTP. The scenarios have been compared synergistically with effluent quality, direct GHG emissions, and energy footprint by the water-energy‑carbon coupling index (WECCI). This modelling study is novel as it fully covers long-term calibration/validation of the model with N2O measurements and tests the dynamic optimisation. Decision-makers and operators can use this new model to optimise GHG emissions and treatment costs.

Unveiling the roles of biofilm in reducing N2O emission in a nitrifying integrated fixed-film activated sludge (IFAS) system.

Biofilm process such as integrated fixed-film activated sludge (IFAS) system has been preliminarily found to produce less nitrous oxide (N2O) than suspended sludge system. However, the N2O emission behaviors and underlying N2O mitigation mechanism in such hybrid system remain unclear. This study therefore aims to fully unveil the roles of biofilm in reducing N2O emission in a nitrifying IFAS system with the aid of some advanced technologies such as N2O microsensor and site-preference analysis. It was found that ammonia oxidation occurred mostly in the sludge flocs (˃ 86%) and biofilm could reduce N2O emission by 43.77% in a typical operating cycle. Biofilm not only reduced nitrite accumulation in nitrification process, inhibiting N2O production via nitrifier denitrification pathway, but also served as a N2O sink, promoting the reduction of N2O via endogenous denitrification. As a result, N2O emissions from the IFAS system were 50%-83% lower than those from the solo sludge flocs. Further, more N2O emission was reduced in the presence of biofilm with decreasing the dissolved oxygen level in the range of 0.5-3.0 mg O2/L. Microbial community and key enzyme analyses revealed that biofilm had relatively high microbial diversity and unique enzyme composition, providing a reasonable explanation for the changed contributions of different N2O production pathways and reduced N2O emission.