Nitrite Denitrification Research Articles

Phycospheric Bacteria Alleviate the Stress of Erythromycin on Auxenochlorella pyrenoidosa by Regulating Nitrogen Metabolism.

Macrolide pollution has attracted a great deal of attention because of its ecotoxic effects on microalgae, but the role of phycospheric bacteria under antibiotic stress remains unclear. This study explored the toxic effects of erythromycin (ERY) on the growth and nitrogen metabolism of Auxenochlorella pyrenoidosa; then, it analyzed and predicted the effects of the composition and ecological function of phycospheric bacteria on microalgae under ERY stress. We found that 0.1, 1.0, and 10 mg/L ERY inhibited the growth and chlorophyll of microalgae, but the microalgae gradually showed enhanced growth abilities over the course of 21 days. As the exposure time progressed, the nitrate reductase activities of the microalgae gradually increased, but remained significantly lower than that of the control group at 21 d. NO3- concentrations in all treatment groups decreased gradually and were consistent with microalgae growth. NO2- concentrations in the three treatment groups were lower than those in the control group during ERY exposure over 21 d. ERY changed the community composition and diversity of phycospheric bacteria. The relative abundance of bacteria, such as unclassified-f-Rhizobiaceae, Mesorhizobium, Sphingopyxis, Aquimonas, and Blastomonas, varied to different degrees. Metabolic functions, such ABC transporters, the microbial metabolism in diverse environments, and the biosynthesis of amino acids, were significantly upregulated in the treatments of higher concentrations (1.0 and 10 mg/L). Higher concentrations of ERY significantly inhibited nitrate denitrification, nitrous oxide denitrification, nitrite denitrification, and nitrite and nitrate respiration. The findings of this study suggest that phycospheric bacteria alleviate antibiotic stress and restore the growth of microalgae by regulating nitrogen metabolism in the exposure system.

Multi-stage anoxic/oxic sequencing batch reactor realizes shortcut nitrogen removal for anaerobically co-digested liquor of municipal sludge and urban organic wastes

ABSTRACT Nitrogen removal from the combined anaerobic digestion dehydration liquor (CADDL) of municipal sludge and urban organic wastes is challenging due to high ammonium concentrations, low C/N ratio, and poor biodegradability. This study proposes a multi-stage anoxic/oxic (A/O) sequencing batch reactor with step feeding to realize partial nitrification and denitrification for shortcut nitrogen removal from the CADDL. We investigated the effects of external carbon source (acetate), dissolved oxygen (DO), A/O duration ratio, and A/O stage number on biological nitrogen removal. Moreover, we assessed the microbial community structure and nitrogen removal pathway. The results showed that the C/N consumption ratio for nitrite reduction to dinitrogen was 3.0 mg COD/mg N, and denitrifying bacteria yielded about 0.43. The optimal dosage of acetate was 2.2 mg COD/mg N. High DO concentration (1.5∼3.0 mg/L) in the aerobic stage improved the ammonia-oxidizing bacteria activity and nitrogen removal rather than worsening the nitritation. A high A/O duration ratio (50 min/60 min) was conducive to complete denitrification of nitrite. The three-stage A/O had an excellent nitrogen removal performance. Under optimal conditions, the nitrite accumulation ratio of nitritation and the total inorganic nitrogen removal reached 100% and 90.1%, respectively. The dominant ammonia-oxidizing bacteria was the genus Nitrosomonas (0.76% abundance), and the dominant denitrifying bacteria was Thauera (0.24% abundance). The nitrite-oxidizing bacteria were not detected, confirming that the biological nitrogen removal pathway was partial nitrification and denitrification. These findings provide a feasible option for the low-carbon nitrogen removal treatment for the CADDL of municipal sludge and urban organic wastes.

Fulvic acid mediated highly efficient heterotrophic nitrification-aerobic denitrification by Paracoccus denitrificans XW11 with reduced C/N ratio

Reducing the C/N ratio requirements for heterotrophic nitrification-aerobic denitrification (HNAD) is crucial for its practical application; however, it remains underexplored. In this study, a highly efficient HNAD bacterium, Paracoccus denitrificans XW11, was isolated. The HNAD characteristics of XW11 were studied, and the redox mediator fulvic acid (FA) was used to reduce the C/N requirements. Whole-genome sequencing revealed multiple denitrification genes in XW11; however, nitrification genes were not identified, because heterotrophic nitrification-related gene sequences were not included in the database. However, the nitrogen removal related enzyme activity test revealed complete nitrification and denitrification pathways. Reverse transcription PCR showed that the membrane-bound nitrate reductase (NarG), rather than the periplasmic nitrate reductase, was responsible for aerobic denitrification. The conventional nitrite reductase (NirS) also does not mediate nitrite denitrification. When the C/N ratio was 10, the ammonia removal efficiency of the Control was 71.71 % and the addition of FA increased it to 86.12 %. Transcriptomic analysis indicated electron flow from the carbon source to FA without proton transmembrane transport, and the presence of FA constructs another electron transfer system. The redox potential of oxidized FA/reduced FA is 0.3679 V, avoiding competition for electrons from Complex III. Thus, ammonia monooxygenase obtains electrons more easily, thereby promoting nitrification. The enzyme activity test of the nitrification process confirmed this view. In addition, NarG expression increased, and the denitrification process was enhanced. Overall, FA improved HNAD efficiency by facilitating electron transfer to the nitrogen dissimilation process, offering a novel approach to reduce the C/N requirement of HNAD.

The effects of polypropylene microplastics on the removal of nitrogen and phosphorus from water by Acorus calamus, Iris tectorum and functional microorganisms

Polypropylene microplastics (PP-MPs), an emerging pollutant, adversely affect the ability of aquatic plants to restore water bodies, thereby compromising the functionality and integrity of wetland ecosystems. This study examines the effects of microplastic stress on the nitrogen and phosphorus removal capacities of Acorus calamus and Iris tectorum, as well as on functional microorganisms within the aquatic system. The findings indicate that under PP-MP stress, the nitrogen and phosphorus absorption capabilities of both plants were diminished. Additionally, there was a significant reduction in the metabolic enzyme activities related to nitrogen and phosphorus in the plants, alongside a notable decrease in leaf nitrogen content. PP-MPs hinder the nutrient uptake of plants, affecting their growth and indirectly reducing their ability to utilize nitrogen and phosphorus. Specifically, in the 10 mg L−1 treatment group, A. calamus and I. tectorum showed reductions in leaf nitrogen content by 23.1% and 31.0%, respectively, and by 14.8% and 27.7% in the 200 mg L−1 treatment group. Furthermore, I. tectorum had higher leaf nitrogen levels than A. calamus. Using fluorescent tagging, the distribution of PP-MPs was traced in the roots, stems, and leaves of the plants, revealing significant growth impairment in both species. This included a considerable decline in photosynthetic pigment synthesis, enhanced oxidative stress responses, and increased lipid peroxidation in cell membranes. PP-MP exposure also significantly reduced the abundance of functional microorganisms involved in denitrification and phosphorus removal at the genus level in aquatic systems. Ecological function predictions revealed a notable decrease in nitrogen cycling functions such as nitrogen respiration and nitrite denitrification among water microorganisms in both treatment groups, with a higher ecological risk potential in the A. calamus treatment group. This study provides new insights into the potential stress mechanisms of PP-MPs on aquatic plants involved in water body remediation and their impacts on wetland ecosystems.

Mature compost promotes biodegradable plastic degradation and reduces greenhouse gas emission during food waste composting

Mature compost can promote the transformation of organic matter (OM) and reduce the emission of polluting gases during composting, which provides a viable approach to reduce the environmental impacts of biodegradable plastics (BPs). This study investigated the impact of mature compost on polybutylene adipate terephthalate (PBAT) degradation, greenhouse gas (GHG) emission, and microbial community structure during composting under two treatments with mature compost (MC) and without (CK). Under MC, visible plastic rupture was advanced from day 14 to day 10, and a more pronounced rupture was observed at the end of composting. Compared with CK, the degradation rate of PBAT in MC was increased by 4.44 % during 21 days of composting. Thermobifida, Ureibacillus, and Bacillus, as indicator species under MC treatment, played an important role in PBAT decomposition. Mature compost reduced the total global warming potential (GWP) by 25.91 % via inhibiting the activity of bacteria related to the production of CH4 and N2O. Functional Annotation of Prokaryotic Taxa (FAPROTAX) further revealed that mature compost addition increased relative abundance of bacteria related to multiple carbon (C) cycle functions such as methylotrophy, hydrocarbon degradation and cellulolysis, inhibited nitrite denitrification and denitrification, thus alleviating the emission of GHGs. Overall, mature compost, as an effective additive, exhibits great potential to simultaneously mitigate BP and GHG secondary pollution in co-composting of food waste and PBAT.

Nitrite Degradation by a Novel Marine Bacterial Strain Pseudomonas aeruginosa DM6: Characterization and Metabolic Pathway Analysis

High concentrations of nitrite in marine aquaculture wastewater not only pose a threat to the survival and immune systems of aquatic organisms but also contribute to eutrophication, thereby impacting the balance of coastal ecosystems. Compared to traditional physical and chemical methods, utilizing microorganism-mediated biological denitrification is a cost-effective and efficient solution. However, the osmotic pressure changes and salt-induced enzyme precipitation in high-salinity seawater aquaculture environments may inhibit the growth and metabolism of freshwater bacterial strains, making it more suitable to select salt-tolerant marine microorganisms for treating nitrite in marine aquaculture wastewater. In this study, a salt-tolerant nitrite-degrading bacterium, designated as DM6, was isolated from the seawater (salinity of 25–30‰) of Portunus trituberculatus cultivation. The molecular identification of strain DM6 was conducted using 16S rRNA gene sequencing technology. The impacts of various environmental factors on the nitrite degradation performance of strain DM6 were investigated through single-factor and orthogonal experiments, with the selected conditions considered to be the key factors affecting the denitrification efficiency of microorganisms in actual wastewater treatment. PCR amplification of key genes involved in the nitrite metabolism pathway of strain DM6 was conducted, including denitrification pathway-related genes narG, narH, narI, nirS, and norB, as well as assimilation pathway-related genes nasC, nasD, nasE, glnA, gltB, gltD, gdhB, and gdhA. The findings indicated that strain DM6 is classified as Pseudomonas aeruginosa and exhibits efficient nitrite degradation even under a salinity of 35‰. The optimal nitrite degradation efficiency of DM6 was observed when using sodium citrate as the carbon source, a C/N ratio of 20, a salinity of 13‰, pH 8.0, and a temperature of 35 °C. Under these conditions, DM6 could completely degrade an initial nitrite concentration of 156.33 ± 1.17 mg/L within 36 h. Additionally, the successful amplification of key genes involved in the nitrite denitrification and assimilation pathways suggests that strain DM6 may possess both denitrification and assimilation pathways for nitrite degradation simultaneously. Compared to freshwater strains, strain DM6 demonstrates higher salt tolerance and exhibits strong nitrite degradation capability even at high concentrations. However, it may be more suitable for application in the treatment of wastewater from marine aquaculture systems during summer, high-temperature, or moderately alkaline conditions.

Evaluation of nitrous oxide emission during ammonia retention from simulated industrial wastewater by microaerobic activated sludge process

Considering the reciprocating processes of nitrogen gas (N2) fixation to ammonia (NH4-N) and NH4-N removal to N2 through nitrification and denitrification during wastewater treatment, a microaerobic activated sludge process (MAS) is proposed in this study as a pretreatment to retain NH4-N from high-strength nitrogenous wastewater for further NH4-N recovery through membrane technology, that is, inhibit nitrification, with sufficient removal of total organic carbon (TOC). With DO and pH control, the 3-reactor bench-scale MAS systems successfully realized an NH4-N retention rate of over 80 %, with TOC removal rates of over 90 %. In addition, the emissions of carbon dioxide (CO2) and nitrous oxide (N2O) during MAS were evaluated. The total N2O emissions were 407 and 475 mg-N/day when pH was controlled at 6.2 (S1) and 6.8 (S2), respectively, with average emission factors to total nitrogen load over 2 % in both systems. Also, the global warming potential of N2O is one order of magnitude larger than that of CO2, indicating the significance of N2O in the MAS process. Therefore, the mechanisms of N2O emission from each reactor were investigated. The first reactor, where most of the TOC was adsorbed, emitted only 1.98 % (S1) and 2.43 % (S2) of the total N2O emissions through the denitrification of nitrite and nitrate (NOx) from the return sludge. The second reactor emitted 79.9 % (S1) and 69.0 % (S2) of the total N2O with the emission rates the same order of magnitude as the NOx production rates. Multiple pathways were considered to contribute to the high N2O emissions, and biotic NH2OH oxidation was one potential pathway at pH 6.2. Finally, the third reactor emitted 9.98 % (S1) and 16.8 % (S2) of the total N2O by nitrifier denitrification. Overall, this study showed that the large N2O emissions under nitrification-inhibiting conditions of the MAS process owed to the incomplete nitrification under acidic conditions and large abundances of denitrifiers. On the other hand, the lower N2O emissions at pH 6.2 evidenced the potential N2O mitigation under slightly more acidic conditions, underlining the necessity of further study on N2O mitigation when adapting to the trend of NH4-N recovery.

Five years nitrogen reduction management shifted soil bacterial community structure and function in high-yielding ‘super’ rice cultivation

Integrated nitrogen (N) management has been adopted for the cultivation of ‘super’ rice to achieve high yield while minimizing environmental risks. How soil microbial communities respond to integrated N management in ‘super’ rice production remains unclear. Five years of field experiment was conducted under a wheat–rice system, with four treatments: conventional farming practices (300 kg ha–1 N), reduced (270 kg ha–1) and increase N (360 kg ha–1) application coupled with increased planting density and accurate irrigation, and a non-N control. The results showed that after five years of treatment, the predominant bacterial phyla shifted from Proteobacteria (22.99%), Acidobacteria (17.04%), and Chloroflexi (14.43%), to Proteobacteria (30.83%), Chloroflexi (20.9%), and Actinobacteria (16.07%). The structure of soil bacterial community differed among the treatments, with available phosphorus contents and pH as key drivers in the first year and NO3--N content in the fifth year. The highest soil N content was detected in the treatment with increased N application, whereas the reduction of N application led to a 32% decrease in soil NO3–-N content. A greater difference was detected in N functional groups in the fifth year than the first year. Following reduced N application, there was also an increased proportion of N-transforming groups, including those involved in aerobic ammonia oxidation, aerobic nitrate oxidation, nitrate denitrification, and nitrite denitrification. Collectively, N fertilizer reduction coupled with accurate irrigation was most effective in regulating soil bacterial communities, especially those associated with N transformation in ‘super’ rice cultivation.

Different grazers and grazing practices alter the growth, soil properties, and rhizosphere soil bacterial communities of Medicago ruthenica in the Qinghai-Tibetan Plateau grassland

Livestock grazing is a popular land-use activity for grasslands. Over grazing and trampling of livestock impairs ecosystem function by changing vegetation growth, soil nutrients, and soil bacterial communities. However, information regarding the characteristics of those allied with above- and belowground of grazing-resistant host plant species under different grazers and grazing practices in grassland ecosystems is limited. We examined the impact of single-species grazing of yak only (YG), sheep only (SG), and mixed grazing in the ratio of yak to sheep of 1:2 (MG (1:2)), 1:4 (MG (1:4)), and 1:6 (MG (1:6)) on plant growth, soil properties, and soil microbial communities. Plant biomass and soil properties were measured in each treatment, and soil microbial communities in the rhizosphere soil of Medicago ruthenica were analyzed by high-throughput sequencing of the 16 S rRNA gene. The results showed that shoot height, root length, and plant FW (fresh weight) and DW (dry weight) of M. ruthenica under single-species grazing decreased by 5.60 %, 3.30 %, 11.34 % and 9.82 %, respectively; under mixed-species grazing decreased by 35.65 %, 29.68 %, 52.35 % and 55.60 %, respectively, compared to CK (no grazing). Mixed-species grazing decreased soil UR (urease), SC (sucrase), and ACP (acid phosphatase) activities by decreasing soil NH4+-N, increasing soil NO3−-N and soil SOC (soil organic carbon), compared to CK. Grazing further affected core phyla and genera of rhizosphere bacterial communities, such as Bacteroidota, Rubrobacter, and Gaiella under different grazers and grazing practices by altering soil properties. The interaction of soil bacteria under YG was more sensitive than that under SG, and the interaction of soil bacteria under mixed-species grazing was more sensitive than that under single-species grazing. According to Functional Annotation of Prokaryotic Taxa (FAPROTAX) analysis associated with N cycling, and the relative abundance of the rhizosphere bacteria with the functions of nitrogen respiration and nitrite denitrification under MG (1:2), MG (1:4), and MG (1:6) were increased by 40.75 %, 34.74 %, 32.31 %, 40.86 %, 36.17 %, and 48.11 %, respectively, compared to CK. This study demonstrates that the mechanism by which different grazers and grazing practices affect soil bacterial communities and associated soil properties in the rhizosphere soil of the legume forage species M. ruthenica will contribute to the understanding how beneficial microbes interact efficiently with grazing-tolerant plant species under different grazing practices.

Nitric oxide and Nitrous oxide accumulation, oxygen production during nitrite denitrification in an anaerobic/anoxic sequencing batch reactor: exploring characteristics and mechanism.

Nitrite denitrification has received increasing attention due to its high efficiency, low energy consumption, and sludge yield. However, the nitric oxide (NO) and nitrous oxide (N2O) which are harmful to the environment, microorganisms, and humans are produced in this process. In order to mitigate NO and N2O production, the biological mechanisms of NO and N2O accumulation, as well as NO detoxification during nitrite denitrification in a sequencing batch reactor were studied. Results showed that the peak of NO accumulation increased from 0.29 [Formula: see text] 0.01 to 3.12 [Formula: see text] 0.34mg L-1 with the increase of carbon to nitrogen ratio (COD/N), which is caused by the sufficient electron donor supply for NO2--N reduction process at high COD/N. Furthermore, the result suggested that NO accumulation with no pH adjustment was 12 times higher than that with pH adjustment. It is related to the inhibition on NO reductase caused by the high free nitrous acid (FNA) and NO concentration with no pH adjustment. The pathways of NO detoxification included NO emission, reduction, and dismutation, and the more NO produced, the high proportion of NO dismutation pathway. Result showed that the maximum of oxygen production during NO dismutation reached to 1.39mg L-1. N2O accumulation was mainly associated with FNA and NO inhibition, COD/N. The peak of N2O accumulation presented a completely opposite trend at pH adjustment and no pH adjustment, it is because that the higher FNA and NO concentration at high COD/N without pH adjustment will inhibit the N2O reductase activity, resulting in the N2O reduction was hindered during nitrite denitrification.

Mathematical modeling of the dynamic effect of denitrifying glycogen-accumulating organisms on nitrous oxide production during denitrifying phosphorus removal

The large amount of intermediate nitrous oxide (N2O) production from denitrifying phosphorus removal (DPR) increases the carbon footprint of wastewater treatment. However, there is a lack of detailed understanding of the interrelationships between denitrifying polyphosphate-accumulating organisms (DPAOs) and denitrifying glycogen-accumulating organisms (DGAOs) on N2O production during DPR. In this work, a mathematical model was developed for the first time to describe dynamic N2O production in the DPR system coexisting DPAOs and DGAOs. The model took into account a four-step subsequent denitrification of nitrate, nitrite, nitric oxide, and N2O. The validity of model was fully tested by comparing simulation studies with experimental data from three independent reports on DPR, which satisfactorily described the dynamics of N2O production, nitrogen oxide reduction, phosphate release and uptake, and intracellular polymers turnover. The validated model could clarify the source and pathways of N2O production. Subsequently, the combined effects of key operational conditions on the overall N2O production, DPAOs and DGAOs competition, and nutrients removal efficiency were investigated by model simulation. Simulation results showed higher polyhydroxyalkanoate storage rate of DGAOs than that of DPAOs during anaerobic stage and the preference of DGAOs for nitrate electron acceptor during anoxic stage. DGAOs dominated the growth competition over DPAOs at low COD (<150 mg/L) and high nitrate (>35 mg/l) conditions, leading to the anoxic storage of glycogen by DGAOs as the main pathway of N2O production. In addition, both N2O production and generation pathways in the system possessed greater variability compared to single DGAOs or DPAOs system, further indicating the necessity of this model.

A quantified nitrogen metabolic network by reaction kinetics and mathematical model in a single-stage microaerobic system treating low COD/TN wastewater

A single-stage intermittent aeration microaerobic reactor (IAMR) has been developed for the cost-effective nitrogen removal from piggery wastewater with a low ratio of chemical oxygen demand (COD) to total nitrogen (TN). In this study, a quantified nitrogen metabolic network was constructed based on the metagenomics, reaction kinetics and mathematical model to provide a revealing insight into the nitrogen removal mechanism in the IAMR. Metagenomics revealed that a complex nitrogen metabolic network, including aerobic ammonia and nitrite oxidation, anammox, denitrification via nitrate and nitrite, and nitrate respiration, existed in the IAMR. A novel method for solving kinetic parameters with high stability was developed based on a genetic algorithm. Use this method to calculate the kinetics of various reactions involved in nitrogen metabolism. Kinetics revealed that simultaneous partial nitritation-anammox (PN/A) and partial denitrification-anammox (PDN/A) were the dominant approaches to nitrogen removal in the IAMR. Finally, a kinetics-based model was proposed for quantitatively describing the nitrogen metabolic network under the limitation of COD. 58% ∼ 67% of nitrogen was removed via the anammox-based processes (PN/A and PDN/A), but only 7% ∼ 12% and 1% ∼ 2% of nitrogen were removed via heterotrophic denitrification of nitrite and nitrate, respectively. The half-inhibition constant of dissolved oxygen (DO) on anammox was simulated as 0.37 ∼ 0.60 mg L−1, filling the gap in quantifying DO inhibition on anammox. High-frequency intermittent aeration was identified as the crucial measure to suppress nitrite-oxidizing bacteria, although it has a high affinity for DO and NO2−-N. In continuous aeration mode, the simulated NO3−-N in the IAMR would rise by 39.6%. The research provides a novel insight into the nitrogen removal mechanism in single-stage microaerobic systems and provides a reliable approach to practicing PN/A and PDN/A for cost-effective nitrogen removal.

Simultaneous nitrification and denitrification in membrane bioreactor: Effect of dissolved oxygen

Membrane bioreactor with the floc activated sludge (mixed liquor suspended solids (MLSS) = 7500 mg/L) was constructed in this work for simultaneously nitrification and denitrification (SND). The effect of dissolved oxygen (DO) on SND process and the nitrogen pathways were investigated. The average TN removal efficiencies were 63.05%, 91.17%, 87.04% and 70.02% for DO 0.5, 1, 2 and 3 mg/L systems, respectively. The effluent ammonia concentration was continuously lower than 5.0 mg/L when the DO was higher than 1 mg/L. Nitrogen in DO 1 and DO 2 mg/L systems was mainly removed via the SND process. The rise of DO concentration increased the abundance of nitrite oxidizing bacteria (NOB) and Nitrospira was the predominant NOB in all the four MBRs. Dechloromonas and Azoarcus were the dominant denitrifying bacteria (DNB) in DO 1 systems responsible for nitrite denitrification. The dominant aerobic DNB Pseudomonas also contributed SND via nitrate denitrification and was little affected by DO changes. Nitrate reductase was the main enzyme for the reduction of NO3−-N to NO2−-N, and narG was the main responsible gene. Nitrite oxidoreductase was the main enzyme for the oxidation of NO2−-N to NO3−-N, and nxrA was the main responsible gene in all the four MBR systems.

Effect of aeration modes on nitrogen removal and N2O emission in the partial nitrification and denitrification process for landfill leachate treatment

The anoxic/multi-aerobic process is widely applied for treating landfill leachate with low carbon to nitrogen ratio. In this study, the effect of two aeration modes in the aerobic phase, i.e. decreasing dissolved oxygen (DO) and increasing DO, on nitrogen removal and N2O emission in the process were systematically compared. The results demonstrate that the aerobic phase with increasing DO mode has a positive effect on improved total nitrogen removal (78 %) under the COD/N ratio as low as 3.45 and minimized N2O emission. DO concentration higher than 1.5 mg/L in the aerobic phase reduced nitrogen removal and led to a significant high N2O emission in the process. Complete nitrite denitrification in the anoxic phase correlated with minimized N2O emission. Under efficient nitrogen removal stage, N2O emission factor was 2.4 ± 1.0 % of the total incoming nitrogen. Microbial analysis revealed that increasing DO mode increased the abundance of ammonia oxidizing bacteria and denitrifiers.

Characterization of Alcaligenes aquatilis as a novel member of heterotrophic nitrifier-aerobic denitrifier and its performance in treating piggery wastewater

A novel strain AS1 with heterotrophic nitrifying-aerobic denitrifying capacity in the species of Alcaligenes aquatilis was isolated from the aerobic activated sludge. It showed a great capability of ammonia removal, and the aerobic metabolic pathways to yield gaseous-nitrogen by hydroxylamine oxidation and nitrite denitrification were proposed. AS1 could efficiently remove ammonia under a wide range of environmental conditions, including the ratio of chemical oxygen demand to total nitrogen: 15–30, pH: 6–10, NaCl: 0–60 g/L, shaking speed of 0–180 rpm, and succinate, acetate, or citrate as carbon source. In the treatment of actual piggery wastewater, 95.3%, 95.1% and 84.9% of NH4+-N was removed by AS1 when the initial ammonia concentration was 500, 1300, and 2000 mg/L, respectively, with the maximum NH4+-N removal rate of 30.5 mg/L/h and 569.7 mg/L/d. Furthermore, plate colony-counting showed that AS1 achieved an efficient proliferation. These results imply the application potential of AS1 in treating high-ammonia wastewater.

Evaluation of nitrogen removal and N2O emission in a novel anammox coupled with sulfite-driven autotrophic denitrification system: Influence of pH

A combination of anaerobic ammonium oxidation (anammox) with sulfur-autotrophic denitrification is a promising alternative for complete nitrogen removal. Herein, a novel anammox coupled with sulfite-driven autotrophic denitrification (ASDAD) is firstly proposed. The influence of pH (8.5–6.5) on nitrogen removal and N2O emission is investigated in an anaerobic sequencing batch biofilm reactor fed with low-nitrogen wastewater containing sulfite. Long-term operation demonstrates that a maximum nitrogen removal of 94.5% occurred at a pH of 7.5 and that the contribution of autotrophic denitrification to nitrogen removal increased as the pH declined. Cyclic and batch experiments verify that anammox and sulfite-autotrophic denitrification simultaneously participates in nitrogen removal, and the stoichiometry for sulfite-driven autotrophic denitrification is firstly proposed. Furthermore, N2O accumulation is also highly correlated with the pH. The N2O emission peaks at a pH of 6.5 due to the inactivated anammox and incomplete autotrophic denitrification of nitrite at such a low pH. High-throughput sequencing reveals the coexistence of anammox (Candidatus Brocadia and Candidatus Kuenenia) and sulfur-oxidizing species (Thiobacillus). Finally, the schema for nitrogen conversion and N2O formation of ASDAD is elucidated based on metagenomic metabolism. This study offers a cost-effective and environment-friendly completely autotrophic nitrogen removal process for municipal wastewater with a low carbon-to-nitrogen ratio in practice.

Inoculation of aerobic granular sludge to achieve granulation under high dissolved oxygen and the associated mechanisms

AGS technology has been studied for decades, and many successful experimental research approaches and full-scale processes have been proposed. However, a precise, comprehensive understanding of the granulation mechanism remains lacking. In this study, sequencing batch reactors were partially inoculated with (S1) or without AGS (S2) to explore aerobic granulation performance in simulative low-strength wastewater. The COD, NH4+-N removal efficiency and SND rate were 91.61%, 92.47%, and 84%, respectively, in S1 and higher in this reactor than in S2 (86.76%, 85.21%, and 75%, respectively), indicating the superiority of partial inoculation with AGS. The ratios of P release/NaAc uptake both increased to nearly 0.250, revealing the robust capacity for phosphorus enrichment. Moreover, the increment of EPS content in S2 (31.73 mg/g MLSS) was 47% higher than that in S1 (21.54 mg/g MLSS) and was contributed mainly by PS, which increased to 30.17 mg/g MLSS (S2). These findings indicated active sludge for granulation was more sensitive to the ambient conditions, and secrete greater PS. FTIR and 3D-EEM revealed that numerous hydroxyl groups were assembled within EPS upon granulation, and humic acid-like substances first increased and then decreased over the operation, implying robust cohesion and positive microbial activities. Microbial community analysis demonstrated that Paracoccus and unclassified_Cytophagales dominated in both reactors, accounting for 29.42% and 10.87%, respectively, of the community in S1 and 26.62% and 18.54%, respectively, of the community in S2. Metabolic pathway analysis revealed enhancement of microbial metabolism, cellular processes, and signaling pathways, such as methanol oxidation, methylotrophy, and nitrite denitrification pathways.

Long-term effects of acetylene on denitrifying N2O production: Biomass performance and microbial community

Maximizing production and use of N2O produced by the process of denitrification has emerged as a promising concept for energy recovery from nitrogen. Acetylene is well known as an inhibitor of N2O reductase activity, however, to date, few studies have utilized long-term cultivation with acetylene to achieve denitrifying N2O recovery. This work assessed the performance of N2O production and shifts in the denitrifying community in response to long-term exposure of acetylene during nitrite denitrification. Batch tests showed that long-term exposure to acetylene at high concentrations (volume fraction >40 %) resulted in high N2O accumulation, up to 98.6 %. High-throughput 16S rRNA gene analysis suggested that Saprospiraceae dominated the microbial community at the end of the cultivation period with its abundance increasing from the initial 3.2% to 42.2%. Metagenomic analysis revealed that after the long-term cultivation under high level acetylene, the relative abundance of nosZ in the community did not change significantly. Instead, the nosZⅡ-harboring bacteria became dominant. Batch tests in series demonstrated that N2O reduction could be restored rapidly upon removal of acetylene. This study describes an effective method for high yield production of N2O during nitrite denitrification that can be used to enhance energy recovery and improves our understanding of the long-term effect of acetylene on the denitrifying community.

Improvement of the performance of simultaneous nitrification denitrification and phosphorus removal (SNDPR) system by nitrite stress

This study investigated a new way to improve the performance of simultaneous nitrification denitrification and phosphorus removal (SNDPR) system by regularly changing the anaerobic/micro-aerobic/anoxic mode to the anaerobic/anoxic mode with 30 mg/L of nitrite dosing. The results indicated that the removal efficiency of total inorganic nitrogen and PO43−-P was improved from 75.44% and 85.14% to 98.89% and 98.17%, respectively. And the good performance of the SNDPR showed a long-time sustainability when the C/N ratio was 5. The results of microbial community illustrated that the abundance of the main nitrite-oxidizing bacteria (NOB), Nitrospira sp., dropped from 5.71% to 0.85% and the abundance of denitrifying polyphosphate-accumulating organisms (DPAOs), Pseudomonas sp. and Acinetobacter sp., increased by 5 times after nitrite stress. The high level of nitric oxide (NO) and free nitrite acid produced by addition of nitrite strongly suppressed the undesired organisms NOB and ordinary heterotrophic denitrifying organisms, and promoted the enrichment of DPAOs. The NO accumulated in the nitrite denitrification process could inhibit NOB and promote AOB. This study revealed that NO plays an important role in regulating the microbial community in the SNDPR system.

Nitrogen isotopic fractionations during nitric oxide production in an agricultural soil

Abstract. Nitric oxide (NO) emissions from agricultural soils play a critical role in atmospheric chemistry and represent an important pathway for loss of reactive nitrogen (N) to the environment. With recent methodological advances, there is growing interest in the natural-abundance N isotopic composition (δ15N) of soil-emitted NO and its utility in providing mechanistic information on soil NO dynamics. However, interpretation of soil δ15N-NO measurements has been impeded by the lack of constraints on the isotopic fractionations associated with NO production and consumption in relevant microbial and chemical reactions. In this study, anoxic (0 % O2), oxic (20 % O2), and hypoxic (0.5 % O2) incubations of an agricultural soil were conducted to quantify the net N isotope effects (15η) for NO production in denitrification, nitrification, and abiotic reactions of nitrite (NO2-) using a newly developed δ15N-NO analysis method. A sodium nitrate (NO3-) containing mass-independent oxygen-17 excess (quantified by a Δ17O notation) and three ammonium (NH4+) fertilizers spanning a δ15N gradient were used in soil incubations to help illuminate the reaction complexity underlying NO yields and δ15N dynamics in a heterogeneous soil environment. We found strong evidence for the prominent role of NO2- re-oxidation under anoxic conditions in controlling the apparent 15η for NO production from NO3- in denitrification (i.e., 49 ‰ to 60 ‰). These results highlight the importance of an under-recognized mechanism for the reversible enzyme NO2- oxidoreductase to control the N isotope distribution between the denitrification products. Through a Δ17O-based modeling of co-occurring denitrification and NO2- re-oxidation, the 15η for NO2- reduction to NO and NO reduction to nitrous oxide (N2O) were constrained to be 15 ‰ to 22 ‰ and −8 ‰ to 2 ‰, respectively. Production of NO in the oxic and hypoxic incubations was contributed by both NH4+ oxidation and NO3- consumption, with both processes having a significantly higher NO yield under O2 stress. Under both oxic and hypoxic conditions, NO production from NH4+ oxidation proceeded with a large 15η (i.e., 55 ‰ to 84 ‰) possibly due to expression of multiple enzyme-level isotopic fractionations during NH4+ oxidation to NO2- that involves NO as either a metabolic byproduct or an obligatory intermediate for NO2- production. Adding NO2- to sterilized soil triggered substantial NO production, with a relatively small 15η (19 ‰). Applying the estimated 15η values to a previous δ15N measurement of in situ soil NOx emission (NOx=NO+NO2) provided promising evidence for the potential of δ15N-NO measurements in revealing NO production pathways. Based on the observational and modeling constraints obtained in this study, we suggest that simultaneous δ15N-NO and δ15N-N2O measurements can lead to unprecedented insights into the sources of and processes controlling NO and N2O emissions from agricultural soils.