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This study presents a new modeling approach for nitrogen recovery in gas-permeable membrane (GPM) contactors, including both ammonia and water transport dynamics. A distinct feature of the model is its capacity to model water transport across the membrane, which has been overlooked in most publications. Osmotic pressure differences are used to predict the behavior of ammonia and water transport in the GPM contactor. Experiments carried out to develop, test and calibrate the model examined the dynamics of ammonia and water transport through the GPM contactor at various nitrogen concentrations. Specifically, the GPM contactor was tested for nitrogen recovery from high-strength synthetic wastewaters (2.4-10.6 g N/L) at 35 °C and at pH 9. The initial volume of the trapping solution (diluted H 2 SO 4 ) was 10 times lower than that of the synthetic wastewater, aiming to concentrate the recovered nitrogen. The estimated ammonia transport constant (K m ) ranged between (1.2 - 2.1)·10 -6 m/s and water transport constant K w between (2.8 - 8.2)·10 -10 m/(s bar). Numerical determination of the model parameters revealed high R² values, demonstrating strong agreement with experimental data., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 The Authors. Published by Elsevier Ltd.. All rights reserved.)

High ammonia stress, which inhibited the performance and stability of anaerobic digestion (AD) systems, is considered a bottleneck problem. To improve the performance of ambient acidogenic AD system under high ammonia stress, three different strategies were developed, including native sludge with nano zero valent iron (nZVI) addition (S nZVI ), domesticated sludge enriched with homoacetogens with no additive (S Domesticated ) and domesticated sludge with nZVI addition (S Domesticated+nZVI ). All groups were operated at ambient temperature (24 ± 1 °C). Results showed that ammonia stress restricted the acidogenic rate in ambient acidogenic system significantly. Under ammonia stress, both S Domesticated and S Domesticated+nZVI showed positive impact on acidogenesis to resist, while the nZVI solely of S nZVI couldn't relieve the ammonia stress effectively. Compared to nZVI or domesticated sludge solely added system, S Domesticated+nZVI showed highest acidogenic rate under high ammonia stress. The S Domesticated treatment increased acetic acid and ethanol production under high ammonia stress compared to the S Control . The S Domesticated+nZVI further increased the production of formic acid and H 2 and reduced the generation of CO 2 . Microbial community analysis indicated that the relative abundances of main acidogens Bifidobacterium, Solobacterium and ethanol producing bacteria Ethanoligenens, increased in the S Domesticated and S Domesticated+nZVI groups. Moreover, S Domesticated+nZVI increased the relative abundance of relevant functional enzyme-encoding genes involved in the generation of acetic acid, formic acid and ethanol and reduced the relative abundance of key functional enzyme-encoding genes related to butyric acid production. This work could provide a novel practical strategy to improve the performance of ambient acidogenic AD system under ammonia stress., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier Ltd. All rights reserved.)

The discovery of complete ammonia oxidizer (comammox) has challenged the traditional understanding of the two-step nitrification process. However, their functions in the oxygen-limited autotrophic nitrification-denitrification (OLAND) process remain unclear. In this study, OLAND was achieved using comammox-dominated nitrifying bacteria in an extremely oxygen-limited environment with a dissolved oxygen concentrations of 0.05 mg/L. The ammonia removal efficiency exceeded 97 %, and the total nitrogen removal efficiency reached 71 % when sodium bicarbonate was used as the carbon source. The pseudo-first- and second-order models were found to best fit the ammonia removal processes under low and high loads, respectively, suggesting distinct ammonia removal pathways. Full-length 16S rRNA gene sequencing and metagenomic results revealed that comammox-dominated under different oxygen levels, in conjunction with anammox and heterotrophic denitrifiers. The abundance of enzymes involved in energy metabolism indicates the coexistence of anammox and autotrophic nitrification-heterotrophic denitrification pathways. The binning results showed that comammox bacteria engaged in horizontal gene transfer with nitrifiers, anammox bacteria, and denitrifiers to adapt to an obligate environments. Therefore, this study demonstrated that comammox, anammox, and heterotrophic denitrifiers play important roles in the OLAND process and provide a reference for further reducing aeration energy in the autotrophic nitrogen removal process., Competing Interests: Declaration of competing interest The authors declare the following financial interests/personal relationships which may be considered as potential competing interests: Yu Xiang reports financial support was provided by National Natural Science Foundation of China. Jia Pu reports financial support was provided by the Natural Science Foundation of Sichuan Province, China. If there are other authors, they declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024. Published by Elsevier Ltd.)

Ammonia (NH 3 ) is an energy vector with an emerging role in decarbonising heat, power and transport through its direct use as a fuel, or indirectly as a hydrogen carrier. Global ammonia production is having to grow to enable the exploitation of NH 3 for energy decarbonisation, which it is projected will consume >50 % of manufacturing capacity by 2050. Ammonia recovered from wastewater can be directly exploited as a sustainable source of ammonia, to reduce the demand for ammonia produced through the energy intensive Haber-Bosch process, while fostering a triple carbon benefit to the water sector, by: (i) avoiding the energy required for aeration of biological processes; (ii) reducing nitrous oxide emissions associated with ammonia oxidation, which is a potent greenhouse gas; and (iii) producing a zero-carbon energy source that can decarbonise energy use. While previous reviews have described technologies relevant for ammonia recovery, to produce ammonia as a zero-carbon fuel or hydrogen carrier, wastewater ammonia must be transformed into the relevant concentration, phase and achieve the product quality demanded for zero carbon heat, power and transport applications, which are distinct from those demanded for more conventional exploitation routes (e.g. agricultural). This review therefore presents a synthesis of established and emerging technologies for the extraction and concentration of ammonia from wastewater, with specific emphasis on enabling the production of ammonia in a form that can be directly exploited for zero carbon energy generation. A précis of technologies for the valorisation of ammonia as a clean energy or hydrogen resource is also introduced, together with discussion of their relevancy and applicability to the water sector including implications to energy, carbon emissions and financial return. The exploitation of ammonia recovered from wastewater as a zero carbon energy source is shown to offer a critical contemporary response for the water sector that seeks to rapidly decarbonise existing infrastructure, while responding to ever stricter nitrogen discharge limits., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 The Author(s). Published by Elsevier Ltd.. All rights reserved.)

Complete ammonia oxidation (comammox) bacteria have been detected in full-scale wastewater treatment plants. However, the slow growth rate limits the research and applications of comammox. Quorum sensing (QS) is an intercellular communication process to regulate microbial physiological metabolism, but its role in comammox Nitrospira has rarely been reported. In this study, denitrifying filter backwash, return and anoxic tank sludges were utilized as seeding inoculums to enrich comammox bacteria, with the addition of three types of signaling molecules (C6-HSL, C8-HSL and C12-HSL). Under ammonia- and dissolved oxygen (DO)-limited conditions, 12 lab-scale sequencing batch reactors (SBRs) were operated for 90 days. Quantitative polymerase chain reaction (qPCR) and 16S rRNA gene sequencing supported the enrichment of comammox Nitrospira in all SBRs. The highest absolute abundance of comammox Nitrospira amoA gene was detected in the anoxic tank sludge with exogenously added C8-HSL, reaching an average of 2.34×10 6 copies/(g sludge). In this condition, comammox Nitrospira and ammonia-oxidizing archaea contributed up to 94 % of the total nitrification activity, with ammonia-oxidizing bacteria accounting for only 6 %. Overall, the role of QS in comammox Nitrospira enrichment was confirmed, with signaling molecules significantly accelerating the growth of comammox Nitrospira, promoting functional enzyme activity, and strengthening nitrifying bacterial competitiveness. Among the signaling molecules tested, C8-HSL exhibited the most pronounced promotional effect, followed by C6-HSL and C12-HSL. Using anoxic tank sludge as the seed sludge with the addition of C8-HSL provides a rapid and reliable enrichment strategy for comammox Nitrospira. These findings offer insights into the role of QS in comammox bacteria enrichment, and might facilitate the development of biotechnologies for wastewater treatment based on comammox Nitrospira., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier Ltd. All rights reserved.)

High ammonia concentration inhibits archaea's activity, causing the accumulation of H 2 and acetate, which suppresses methane production in anaerobic digestion (AD). The study aimed to enhance microbial hydrogen metabolism through a side-stream hydrogen domestication (SHD) strategy, which involves applying hydrogen stimulation to a portion of the sludge separately. SHD maintained a stable methane yield of 407.5 mL/g VS at a high total ammonia nitrogen (TAN) concentration of 3.1 g/L. In contrast, the control group gradually decreased and stopped methane production at a TAN concentration of 2.3 g/L. Further analysis using enzyme activity assays, flow cytometry, and metagenomics explored the mechanisms underlying ammonia tolerance of SHD-treated group. SHD reshaped the microbial community, enriching homoacetogens and Methanosaeta-dominated methanogenic archaea. Key metabolic pathways including homoacetogenesis, butyrate degradation, propionate degradation, and methane production were enhanced. The activity of related enzymes also increased. Gene abundance in energy-generating pathways, such as glycolysis, was enhanced, ensuring adequate ATP production. Additionally, the high gene abundance of ion transport systems contributed to regulating proton imbalance and supplementing intracellular K + . This study provides important insights and practical guidance for developing novel techniques in the field of anaerobic digestion., Competing Interests: Declaration of competing interest The authors declare that they have no known competing interests or personal relationships that could have appeared to influence the work reported in this paper and all authors have seen and approved the final version of the manuscript. They warrant that the article is the authors’ original work, has not been published prior and is not under consideration for publication elsewhere., (Copyright © 2024. Published by Elsevier Ltd.)

The addition of conductive materials (CMs) is an effective strategy for mitigating ammonia inhibition during anaerobic digestion (AD). However, the introduction of CMs can result in increased antibiotic resistance genes (ARGs) pollution, potentially facilitated by enhanced horizontal gene transfer (HGT). The complex dynamics of intracellular and extracellular ARGs (iARGs/eARGs) and the mechanisms underlying their transfer, mediated by CMs, in ammonia-stressed AD systems remain unclear. In this study, we investigated the effects of three commonly used CMs-nano magnetite (Mag), nano zero-valent iron (nZVI), and granular activated carbon (GAC)-on the fate of iARGs and eARGs during the AD of waste activated sludge under ammonia stress. The results revealed an unexpected enrichment of iARGs by 1.5 %-10.9 % and a reduction of eARGs by 14.1 %-25.2 % in CM-supplemented AD. This discrepancy in the dynamics of iARGs and eARGs may be attributed to changes in microbial hosts and the horizontal transfer of ARGs. Notably, CMs activated prophages within antibiotic-resistant bacteria (ARB) and their symbiotic partners involved in vitamin B12 provision, leading to the lysis of ARB and the subsequent release of eARGs for transformation. Additionally, the abundance of potentially mobile ARGs, which co-occurred with mobile genetic elements, increased by 56.6 %-134.5 % with CM addition, highlighting an enhanced potential for the HGT of ARGs. Specifically, Mag appeared to promote both transformation and conjugation processes, while nZVI only promoted conjugation. Moreover, none of the three CMs had any discernible impact on transduction. GAC proved superior to both nano Mag and nZVI in controlling the enrichment of iARGs, reducing eARGs, and limiting HGTs simultaneously. Overall, these findings provide novel insights into the role of viruses and the mechanisms of ARG spread in CM-assisted AD, offering valuable information for developing strategies to mitigate ARG pollution in practical applications., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier Ltd. All rights reserved.)

Microbial chain elongation integrating innovative bioconversion technologies with organic waste utilization can transition current energy-intensive n-caproic acid production to sustainable circular bioeconomy systems. However, ammonia-rich waste streams, despite their suitability, pose inhibitory challenges to these bioconversion processes. Herein, biochar was employed as an additive to enhance the activity of chain elongating microbes under ammonia inhibition conditions, with an objective to detail underlying mechanisms of improvements. Biochar addition significantly improved chain elongation performance even under severe ammonia stress (exceeding 8 g N/L), increasing n-caproic acid yields by 40 % to 158 % and reducing lag times by 51 % to 90 %, compared with the best-performing group without biochar addition. The material contribution to n-caproic production reached up to 94.3 % (at 4 g N/L). These enhancements were mainly attributed to the new electron syntrophy induced by biochar, which improved electron transfer system activity and electrical conductivity of the fermentation system. This is further substantiated by increased relative abundances of the genus Sporanaerobacter, electroactive bacteria, and up-regulated direct electron transfer-related genes including conductive pili and c-type cytochrome. This study demonstrates that biochar can confer robust resilience to ammonia toxicity in functional microbes, paving a way for efficient and sustainable n-caproic acid production., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier Ltd. All rights reserved.)

The advanced oxidation process is an efficient technology for the degradation and detoxification of refractory organics to ensure water safety. However, most researches focus on improving pollutant degradation but overlook carbon emission and resource utilization. In this study, a flow-through electrochemical integrated system was constructed to simultaneously realize bisphenol A (BPA) oxidation into small non-toxic organics and CO 2 , and generated CO 2 coupled with nitrate-containing wastewater conversion to urea and ammonia on a porous cathode (Zr-Fe/CN). The synergistic effect between anodic BPA oxidation with cathodic CO 2 and NO 3 - reduction improves the electron utilization efficiency and thus increasing the BPA degradation, urea yield rate (UYR) and NH 3 yield rate (NYR) by 13.4 % 18.4 % and 8.3 %, respectively. Furthermore, the flow-through operation mode significantly increased the mass transfer efficiency and quickly carried generated CO 2 from the anode into the cathode to improve CO 2 utilization efficiency. Compared to the parallel plate electrode reactor, the BPA degradation efficiency, UYR and NYR in the flow-through reactor increased from 59.46 % to 84.49 % (the initial concentration of BPA was 40 mg/L), 9.94 mmol h -1 g -1 to 19.55 mmol h -1 g -1 , and 80.31 mmol h -1 g -1 to 106.06 mmol h -1 g -1 within 60 min, respectively. Moreover, the total carbon conversion efficiency (from BPA to urea) increased from 20.2 % to 42.4 % and the total Faraday efficiency (FE) increased from 78.6 % to 96.3 %. This work provides a multi-win strategy of harmless, resource-based and carbon emission reduction for wastewater treatment., Competing Interests: Declaration of competing interest The authors declare the following financial interests/personal relationships which may be considered as potential competing interests: Ying Wang reports financial support was provided by National Natural Science Foundation of China. If there are other authors, they declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier Ltd. All rights reserved.)

The mainstream partial nitritation/anammox (PN/A) process represents a significant innovation in decarbonizing municipal wastewater treatment. However, its implementation is considerably hampered by the challenge of stable nitrite supply. In this study, a pilot-scale PN/A system receiving real sewage (20 m 3 ) was operated at room temperature for nearly one year. Remarkable PN performance with relatively high nitrite accumulation ratio of 75.04 ± 10.05 % was obtained via in-situ free ammonia (FA) strategy. The ammonium concentration enriched in the zeolite increased significantly by 548.8 times compared to that in the aqueous phase by ion exchange. This substantial increase robustly inhibited nitrite-oxidizing bacteria (NOB), resulting in high relative abundance ratio of ammonia-oxidizing bacteria (AOB) to NOB of 37.93 ± 12.61 in the zeolite biofilm, compared to 10.22 ± 1.67 in suspended floc sludge. The significant differences in FA concentrations between zeolite biofilm and suspended floc sludge resulted in distinct spatial distribution disparities of AOB and NOB, which were central to achieving stable nitrite accumulation without complex multiple selective pressures. Consequently, compliant effluent with total nitrogen of 10.91 ± 4.23 mg N/L was achieved at 10.4-31.1 °C without external carbon source addition. The biocarriers in the anammox process played a key role in enhancing functional genes and electron flow, supporting anammox-dominated nitrogen removal. This study presents a flexible and adaptable strategy for mainstream nitrite shunting, highlighting its potential for large-scale implementation of mainstream anammox treatment., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier Ltd. All rights reserved.)

While air stripping combined with acid scrubbing remains a competitive technology for the removal and recovery of ammonia from wastewater streams, its use of strong acids is concerning. Organic acids offer promising alternatives to strong acids like sulphuric acid, but their application remains limited due to high cost. This study proposes an integration of air stripping and organic acid scrubbing with bipolar membrane electrodialysis (BPMED) to regenerate the organic acids. We compared the energy consumption and current efficiency of BPMED in recovering dissolved ammonia and regenerating sulphuric, citric, and maleic acids from synthetic scrubber effluents. Current efficiency was lower when regenerating sulphuric acid (22 %) compared to citric (47 %) and maleic acid (37 %), attributable to the competitive proton transport over ammonium across the cation exchange membrane. Organic salts functioned as buffers, reducing the concentration of free protons, resulting in higher ammonium removal efficiencies with citrate (75 %) and malate (68 %), compared to sulphate (29 %). Consequently, the energy consumption of the BPMED decreased by 54 % and 35 % while regenerating citric and maleic acids, respectively, compared to sulfuric acid. Membrane characterisation experiments showed that the electrical conductivity ranking, ammonium citrate > ammonium malate > ammonium sulphate, was mirrored by the energy consumption (kWh/kg-N recovered) ranking, ammonium sulphate (15.6) < ammonium malate (10.2) < ammonium citrate (7.2), while the permselectivity ranking, ammonium sulphate > ammonium citrate > ammonium malate, aligned with calculated charge densities. This work demonstrates the potential of combining organic acid scrubbers with BPMED for ammonium recovery from wastewater effluents with minimum chemical input., Competing Interests: Declaration of competing interest The authors declare the following financial interests/personal relationships which may be considered as potential competing interests: Gladys Mutahi reports financial support was provided by Netherlands Enterprise Agency. If there are other authors, they declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 The Author(s). Published by Elsevier Ltd.. All rights reserved.)

Perfluorooctanoic acid (PFOA), benzalkyl dimethylammonium compounds (BAC) and antibiotic resistance genes (ARGs) have negative effects on biological sewage treatment. The performance of nitrification systems under stress of PFOA (0.1-5 mg/L) or/and BAC (0.2-10 mg/L) was explored during 84-day experiments using four sequencing batch reactors, in this study. Low (0.1 mg/L) concentration PFOA had a positive influence on ammonia removal, while medium and high (2 and 5 mg/L) concentrations PFOA caused severe inhibition. Meanwhile, PFOA stress resulted in the enrichment of ARGs in water (w-ARGs). BAC (0-10 mg/L) had no obvious influence on ammonia removal. However, BAC promoted the reduction of ARGs and the bacterial community was the main participator (48.07%) for the spread of ARGs. Interestingly, the joint stress of PFOA and BAC increased the ammonia-oxidizing bacteria (AOB) activity from 5.81 ± 0.19 and 6.05 ± 0.79 mg N/(g MLSS·h) to 7.09 ± 0.87 and 7.23 ± 0.29 mg N/(g MLSS·h) in medium and high concentrations, compared to single stress of PFOA, which was observed for the first time. BAC could reduce bioavailability of PFOA through competitive adsorption and decreasing sludge hydrophobicity by the lower β-Sheet and α-Helix in tightly bound protein. Furthermore, the joint stress of PFOA and BAC was able to intensify the proliferation of w-ARGs and extracellular ARGs in sludge, and developed the most active horizontal gene transfer mediated by intl1 compared to single stress of PFOA or BAC. The batch tests verified the detoxification capacity of BAC on nitrification under 2.5 mg/L PFOA (48 h exposing), and the maximum alleviation of AOB activity was achieved at BAC and PFOA mass ratio of 2:1. In summary, BAC could be used to alleviate the inhibition of PFOA on ammonia oxidation, providing an efficient and sustainable approach in wastewater treatment., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier Ltd. All rights reserved.)

How to intensify the ammonia oxidation rate (AOR) is still a bottleneck impeding the technology development for the innovative acidic partial nitritation because the eosinophilic ammonia-oxidizing bacteria (AOB), such as Nitrosoglobus or Nitrosospira, were inhibited by the high-level free nitrous acid (FNA) accumulation in acidic environments. In this study, an innovative approach of dynamic acidic pH regulation control strategy was proposed to realize high-rate acidic partial nitritation driven by common AOB genus Nitrosomonas. The acidic partial nitrification process was carried out in a laboratory-scale sequencing batch moving bed biofilm reactor (SBMBBR) for long-term (700 days) to track the effect of dynamic acidic pH on nitrifying bacterial activity. The results indicated that the influent NH 4 + -N concentration was about 100 mg/L, the nitrite accumulation ratio was exceeding 90%, and the maximum AOR can reach 14.5 ± 2.6 mg N L -1 h -1 . Although the half-saturation inhibition constant of NOB (K I&#95;FNA(AOB) ) reached 0.37 ± 0.10 mg HNO 2 N/L and showed extreme adaptability in FNA, the inactivation effect of FNA (6.1 mg HNO 2 N/L) for NOB was much greater than that of AOB, with inactivation rates of 0.61 ± 0.08 h -1 , respectively. The effluent pH was gradually reduced to 4.5 by ammonia oxidation process and the periodic FNA concentration reached 6.5 mg HNO -1 , respectively. The effluent pH was gradually reduced to 4.5 by ammonia oxidation process and the periodic FNA concentration reached 6.5 mg HNO 2 N/L to inactivate nitrite-oxidizing bacteria (NOB) without negatively affecting Nitrosomonas during long-term operation. This result provides new insights for the future implementation of high-rate stabilized acidic partial nitritation by Nitrosomonas., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier Ltd. All rights reserved.)

This study investigated the impact of varying total ammonia nitrogen (TAN) feed levels along with water temperature decreases on the performance of nitrifying moving bed biofilm reactor (MBBR) at 1 °C and its recovery at 3 °C. Five MBBR reactors were operated with different TAN concentrations as water temperature decreased from 20 to 3 °C: reactor R1 at 30 mg N/L, reactor R2 at 20 mg N/L, reactor R3 at 15 mg N/L, reactor R4 at 10 mg N/L and reactor R5 at 0 mg N/L. The corresponding biofilm characteristics were also analyzed to understand further nitrifying MBBR under different TAN feeding scenarios. The findings revealed that the higher TAN levels were before reaching 1 °C, the better nitrification performance and the more biomass grew. However, the highest TAN concentration (30 mg N/L) might negatively affect the nitrification performance, the activity of nitrifiers, and the growth of biofilms at 1 °C because of the toxic effects of un-ionized or free ammonia (FA). It was observed that the activities of ammonia-oxidizing bacteria (AOB) and nitrite-oxidizing bacteria (NOB) were affected by FA concentrations ranging from 0.2 to 0.7 mg N/L at 1 °C, but they could gradually be adapted to such inhibitory environment, with NOB recovering more quickly and robustly than AOB. The study identified 20 mg N/L (67 % of maximum influent TAN at 1 °C in R2 as the optimal TAN feeding concentration, achieving over 90 % TAN removal and a surface area removal rate (SARR) of 0.78 ± 0.02 g N/m 2 ·d at 1 °C. Meanwhile, R2 also exhibited the highest biofilm mass, with total solids at 13.3 mg/carrier and volatile solids at 11.3 mg/carrier. As TAN was removed, nitrite accumulation was observed at 1 °C, and higher influent TAN concentrations prior to 1 °C appeared to delay the accumulation. When water temperature increased from 1 °C to 3 °C, nitrification performance improved significantly in all reactors without nitrite accumulation, and the higher TAN feeding in the previous stage led to faster recovery. Compared with 20 °C, biofilm became thinner and denser at 1 °C and 3 °C. Furthermore, this study revealed significant shifts in microbial community composition and nitrifier abundances in response to changes in water temperature and influent TAN levels. The dominant nitrifiers were identified as Nitrosomonadaceae (AOB) and Nitrospiraceae (NOB). At 1 °C, the nitrifier abundances were significantly correlated with SARRs, FA, and biofilm density. R2, which exhibited the best nitrification performance, maintained higher nitrifier abundances at 1 °C., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier Ltd. All rights reserved.)

This study aimed to investigate whether separating organics depletion from nitrification increases the overall performance of urine nitrification. Separate organics depletion was facilitated with membrane aerated biofilm reactors (MABRs). The high pH and ammonia concentration in stored urine inhibited nitrification in the first stage and therewith allowed the separation of organics depletion from nitrification. An organics removal of 70 % was achieved at organic loading rates in the influent of 3.7 g COD d -1 m -2 . Organics depletion in a continuous flow stirred tank reactor (CSTR) for organics depletion led to ammonia stripping through diffused aeration of up to 13 %. Using an MABR, diffusion into the lumen amounted for 4 % ammonia loss only. In the MABR, headspace volume and therefore ammonia loss through the headspace was negligible. By aerating the downstream MABR for nitrification with the off-gas of the MABR for organics depletion, 96 % of the ammonia stripped in the first stage could be recovered in the second stage, so that the overall ammonia loss was negligibly low. Nitrification of the organics-depleted urine was studied in MABRs, CSTRs, and sequencing batch reactors in fed batch mode (FBRs), the latter two operated with suspended biomass. The experiments demonstrated that upstream organics depletion can double the nitrification rate. In a laboratory-scale MABR, nitrification rates were recorded of up to 830 mg N L -1 d -1 (3.1 g N m -2 d -1 ) with ambient air and over 1500 mg N L -1 d -1 (6.7 g N m -2 d -1 ) with oxygen-enriched air. Experiments with a laboratory-scale MABR showed that increasing operational parameters such as pH, recirculation flow, scouring frequency, and oxygen content increased the nitrification rate. The nitrification in the MABR was robust even at high pH setpoints of 6.9 and was robust against process failures arising from operational mistakes. The hydraulic retention time (HRT) required for nitrification was only 1 to 2 days. With the preceding organics depletion, the HRT for our system requires 2 to 3 days in total, whereas a combined activated sludge system requires 4 to 8 days. The N 2 O concentration in the off-gas increases with increasing nitrification rates; however, the N 2 O emission factor was 2.8 % on average and independent of nitrification rates. These results indicate that the MABR technology has a high potential for efficient and robust production of ammonium nitrate from source-separated urine., Competing Interests: Declaration of competing interest The authors declare the following financial interests/personal relationships which may be considered as potential competing interests: Kai M. Udert reports a relationship with VunaNexus that includes: board membership and travel reimbursement. VunaNexus uses biological and physical processes for nutrient recovery from urine. The study was not influenced by the relationship of Kai M. Udert with VunaNexus. Aurea Heusser and Kai M. Udert have a patent pending to Eawag. The patent includes the separate removal of organics from source-separated urine, intermediate pharmaceutical removal by adsorption and subsequent nitrification. The other authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 The Author(s). Published by Elsevier Ltd.. All rights reserved.)

Ammonia-oxidizing bacteria (AOB) are ubiquitous on the earth and have broad applications in bioremediation. However, the number of their species with standing in nomenclature and deposited in Microbial Culture Collections still remains low. Moreover, only a few novel species have been reported over the last decades. In this study, we sealed agar in serum bottles to develop a kind of solid agar plate with the oxygen concentration in the headspace maintained at low levels. By using these plates, eight AOB isolates including two novel species were obtained. When AOB cells were grown on the sealed solid agar plates, the time to form visible colonies was largely reduced and the maximum diameter of colonies reached 2 mm, which makes the process of AOB isolation rapid and efficient. Based on five AOB isolates, the headspace oxygen concentration had a significant influence on AOB growth either on solid plate or in liquid culture. Especially, when grown under 21 % O 2 , the number of colonies formed on solid agar plates was very low and sometimes no visible colony formed. Besides the application on AOB isolation, the sealed solid agar plate was also effective for the enumeration and preservation of AOB cells. When preserved under room temperature for more than ten months, the AOB colonies on the plate could still be recovered. This method provides a feasible way to isolate more novel AOB species from the environment and deposit more species in Microbial Culture Collections., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier Ltd. All rights reserved.)

Anthropogenic activities significantly impact the elemental cycles in aquatic ecosystems, with the N-cycling playing a critical role in potential nutrient turnover and substance cycling. We hypothesized that measures to prevent COVID-19 transmission profoundly altered the nitrogen cycle in riverine ecosystems. To investigate this, we re-analyzed metagenomic data and identified 60 N-cycling genes and 21 host metagenomes from four urban reaches (one upstream city, Wuhan, and two downstream cities) along the Yangtze River. Our analyses revealed a marked decrease in the abundance of bacterial ammonia monooxygenase genes, as well as in the host, ammonia-oxidizing autotrophic Nitrosomonas, followed by a substantial recovery post-pandemic. We posited that discharge of sodium hypochlorite (NaOCl) disinfectant may be a primary factor in the reduction of N-cycling process. To test this hypothesis, we exposed pure cultures of Nitrosomonas europaea to NaOCl to explore the microbial stress response. Results indicated that NaOCl exposure rapidly compromised the cell structure and inhibited ammonia oxidation of N. europaea, likely due to oxidative stress damage and reduced expression of nitrogen metabolism-related ammonia monooxygenase. Using the functional tagging technique, we determined that NaOCl directly destroyed the ammonia monooxygenase protein and DNA structure. This study highlights the negative impacts of chlorine disinfectants on the function of aquatic ecosystems and elucidates potential mechanisms of action., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024. Published by Elsevier Ltd.)

Ammonia recovery from wastewater is of great significance for aquatic ecology safety, human health and carbon emissions reduction. Electrochemical methods have gained increasing attention since the authigenic base and acid of electrochemical systems can be used as stripper and absorbent for transmembrane chemisorption of ammonia, respectively. However, the separation of electrodes and gas permeable membrane (GPM) significantly restricts the ammonia transfer-transformation process and the authigenic acid-base utilization. To break the restrictions, this study developed a gas permeable membrane electrode assembly (GPMEA), which innovatively integrated anode and cathode on each side of GPM through easy phase inversion of polyvinylidene fluoride binder, respectively. With the GPMEA assembled in a stacked transmembrane electro-chemisorption (sTMECS) system, in situ utilization of authigenic acid and base for transmembrane electro-chemisorption of ammonia was achieved to enhance the ammonia recovery from wastewater. At current density of 60 A/m 2 , the transmembrane ammonia flux of the GPMEA was 693.0 ± 15.0 g N/(m 2 ·d), which was 86 % and 28 % higher than those of separate GPM and membrane cathode, respectively. The specific energy consumption of the GPMEA was 9.7∼16.1 kWh/kg N, which were about 50 % and 25 % lower than that of separate GPM and membrane cathode, respectively. Moreover, the application of GPMEA in the ammonia recovery from wastewater is easy to scale up in the sTMECS system. Accordingly, with the features of excellent performance, energy saving and easy scale-up, the GPMEA showed good prospects in electrochemical ammonia recovery from wastewater., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024. Published by Elsevier Ltd.)

This study developed a new process that stably produced ammonium nitrate (NH 4 NO 3 ), an important and commonly used fertilizer, from the source-separated urine by comammox Nitrospira. In the first stage, the complete conversion of ammonium to nitrate was achieved by comammox Nitrospira. In this scenario, the pH was maintained at 6 by adding external alkali, which also provided sufficient alkalinity for full nitrification. In the second stage, the NH 4 NO 3 was produced directly by comammox Nitropsira by converting half of the ammonium in urine into nitrate. In this case, no alkali was added and pH automatically dropped and self-maintained at an extremely acidic level (pH 3-4). In both scenarios, negligible nitrite accumulation was observed, while the final product of the second stage contained ammonium and nitrate at the molar ratio of 1:1. The dominance of comammox Nitrospira over canonical ammonia-oxidizing bacteria (AOB) and nitrite-oxidizing bacteria (NOB) was systematically proved by the combination of 16S rRNA gene amplicon sequencing, quantitative polymerase chain reaction, and metagenomics. Notably, metagenomic sequencing suggested that the relative abundance of comammox Nitrospira was over 20 % under the acidic condition at pH 3-4, while canonical AOB and NOB were undetectable. Batch experiments showed that the optimal pH for the enriched comammox Nitrospira was ∼7, which could sustain their activity in a wider pH range from 4 to 8 surprisingly but lost activity at pH 3 and 9. The findings not only present an application potential of comammox Nitrospira in nitrogen recovery from urine wastewater but also report the survivability of comammox bacteria in acidic environments., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 The Authors. Published by Elsevier Ltd.. All rights reserved.)

The ammonia recovery from wastewater via electrochemical technologies represents a promising way for wastewater treatment, resource recovery, and carbon emissions reduction. However, chemicals consumption and reactors scalability of the existing electrochemical systems have become the key challenges for their development and application. In this study, a stacked transmembrane electro-chemisorption (sTMECS) system was developed to utilize authigenic acid and base on site for enhancing ammonia recovery from wastewater. The easily scaled up system was achieved via innovatively connecting the cathode chamber in a unit with the anode chamber in the adjacent unit by a hydrophobic gas permeable membrane (GPM). Thus, authigenic base at cathodes and authigenic acid at anodes could be utilized as stripper and absorbent on site to enhance the transmembrane chemisorption of ammonia. Continuous power supply, reducing the distances of electrodes to GPM and moderate aeration of the catholyte could promote ammonia recovery. Applied to the ammonia recovery from the simulated urine, the sTMECS under the current density 62.5 A/cm 2 with a catholyte aeration rate of 3.2 L/(L⋅min) for operation time 4 h showed the transmembrane ammonia flux of 26.00 g N/(m 2 ·h) and the system energy consumption of 10.5 kWh/kg N. Accordingly, the developed sTMECS system with chemicals saving, easy scale-up and excellent performance shows good prospects in recovering ammonia from wastewater., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier Ltd. All rights reserved.)

The global energy crisis has intensified the search for sustainable and clean alternatives, with biohydrogen emerging as a promising solution to address environmental challenges. Leveraging photo fermentation (PF) process, purple phototrophic bacteria (PPB) can harness reducing power derived from organic substrates to facilitate hydrogen production. However, existing studies report much lower H 2 yields than theoretical value when using acetate as carbon source and ammonia as nitrogen source, primarily attributed to the widely employed pulse-feeding mode which suffers from ammonia inhibition effect on nitrogenase. To address this issue, a continuous feeding mode was applied to avoid ammonia accumulation in this study. On the other hand, other pathways like carbon fixation and polyhydroxyalkanoate (PHA) formation could compete reducing power with H 2 production. However, the reducing power allocation under continuous feeding mode is not yet clear. In this study, the reducing power allocation and hydrogen production performance were evaluated under various ammonia loading, using acetate as carbon source and infrared LED at around 50 W·m -2 as light source. The results show that (a) The absence of ammonia resulted in the best performance for hydrogen production, with 44 % of the reducing power distributed to H 2 and the highest H 2 volumetric productivity, while the allocation of reducing power to hydrogen production stopped when ammonia loading was above 7.6 mg NH 4 -N·L -1 ·d -1 ; (b) when PPB required to eliminate reducing power under ammonia limited conditions, PHA production was the preferred pathway followed by the hydrogen production pathway, but once PHA accumulation reached saturation, hydrogen generation pathway dominated; (c) under ammonia limited conditions, the TCA cycle was more activated rendering higher NADH (i.e. reducing power) production compared with that under ammonia sufficient conditions which was verified by metagenomics analysis, and all the hydrogen production, PHA accumulation and carbon fixation pathways were highly active to dissipate reducing power. This work provides the insight of reducing power distribution and PPB biohydrogen production variated by ammonia loading under continuous feeding mode., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier Ltd. All rights reserved.)

A novel integrated pilot-scale A-stage high rate activated sludge, B-stage short-cut biological nitrogen removal and side-stream enhanced biological phosphorus removal (A/B-shortcut N-S2EBPR) process for treating municipal wastewater was demonstrated with the aim to achieve simultaneous and carbon- and energy-efficient N and P removal. In this studied period, an average of 7.62 ± 2.17 mg-N/L nitrite accumulation was achieved through atypical partial nitrification without canonical known NOB out-selection. Network analysis confirms the central hub of microbial community as Nitrospira, which was one to two orders of magnitude higher than canonical aerobic oxidizing bacteria (AOB) in a B-stage nitrification tank. The contribution of comammox Nitrospira as AOB was evidenced by the increased amoB/nxr ratio and higher ammonia oxidation activity. Furthermore, oligotyping analysis of Nitrospira revealed two dominant sub-clusters (microdiveristy) within the Nitrospira. The relative abundance of oligotype II, which is phylogenetically close to Nitrospira&#95;midas&#95;s&#95;31566, exhibited a positive correlation with nitrite accumulation in the same operational period, suggesting its role as comammox Nitrospira. Additionally, the phylogenetic investigation suggested that heterotrophic organisms from the family Comamonadacea and the order Rhodocyclaceae embedding ammonia monooxygenase and hydroxylamine oxidase may function as heterotrophic nitrifiers. This is the first study that elucidated the impact of integrating the S2EBPR on nitrifying populations with implications on short-cut N removal. The unique conditions in the side-stream reactor, such as low ORP, favorable VFA concentrations and composition, seemed to exert different selective forces on nitrifying populations from those in conventional biological nutrient removal processes. The results provide new insights for integrating EBPR with short-cut N removal process for mainstream wastewater treatment., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024. Published by Elsevier Ltd.)

Recovering ammonia from waste streams (e.g., urine) is highly desirable to reduce natural gas-based NH 3 production and nitrogen discharge into the water environment. Electrochemical membrane stripping is an attractive alternative because it can drive NH 4 + transformation to NH 3 production; however, the conventional configurations suffer from relatively low ammonia recovery (<80 %) and significant acid/material usage for ammonia adsorption. To this end, we develop a novel stack system that simply uses an oxygen evolution reaction to in-situ produce acid from water, enabling chemical-free ammonia recovery from synthetic urine. In batch mode, the percentage removal and recovery increased respectively from 74.5 % to 97.9 % and 81.8 % to 92.7 % when the electrode pairs increased from 2 to 4 in the stack system. To address the gas-sparging issue that deteriorated ammonia recovery in continuous operation, pulsed electric field (PEF) mode was applied, resulting in ∼100 % recovery under optimized conditions. At an ammonia removal rate of 35.1 g-N m - production; however, the conventional configurations suffer from relatively low ammonia recovery (<80 %) and significant acid/material usage for ammonia adsorption. To this end, we develop a novel stack system that simply uses an oxygen evolution reaction to in-situ produce acid from water, enabling chemical-free ammonia recovery from synthetic urine. In batch mode, the percentage removal and recovery increased respectively from 74.5 % to 97.9 % and 81.8 % to 92.7 % when the electrode pairs increased from 2 to 4 in the stack system. To address the gas-sparging issue that deteriorated ammonia recovery in continuous operation, pulsed electric field (PEF) mode was applied, resulting in ∼100 % recovery under optimized conditions. At an ammonia removal rate of 35.1 g-N m -2  h -1 , our chemical-free system in PEF mode has achieved significantly higher ammonia recovery (>90 %) from synthetic urine. The total cost to recover 1 kg of NH -1 -N from real human urine was $15.9 in the proposed system. Results of this study demonstrate that this novel approach holds great promise for high ammonia recovery from waste streams, opening a new pathway toward sustainable nitrogen management.3 -N from real human urine was $15.9 in the proposed system. Results of this study demonstrate that this novel approach holds great promise for high ammonia recovery from waste streams, opening a new pathway toward sustainable nitrogen management., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier Ltd. All rights reserved.)

The removal and recovery of ammonia from wastewater are critical processes for achieving global environmental sustainability and promoting circular economic development. High-gravity technology is an advanced solution to achieve ammonia stripping from wastewater. This study used machine-learning (ML) techniques to provide more comprehensive insights on various influencing factors, including the operating parameters, wastewater characteristics, and design parameters of rotating packed beds. Bayesian auto-optimization combined with a boosting algorithm effectively overcame the challenges of modeling complex datasets with small sample sizes, multidimensional data, missing values, and skewed distributions. Accurate ML based predictive models for the ammonia removal efficiency (η) and mass transfer coefficient (K L a) were developed, the performance on the training set was R 2  = 0.98 and R 2  = 0.89, and on the testing set was R 2  = 0.98 and R 2  = 0.82. The developed model revealed that the stripping stage and gas-liquid ratio were the most influential features for predicting η, whereas the liquid flow and high-gravity factor were the most important features for predicting K L a. The well-trained model was then deployed in an online software application that could provide both predictive and auto-update functions for operators and managers, ensuring that practitioners could use the model. The end-to-end machine-learning approach used in this study-that is, covering data collection, model development, and application-could improve the availability of research results, providing valuable references for the further advancement of technology in the field of environmental., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2023. Published by Elsevier Ltd.)

Vacuum thermal stripping permits the recovery of ammonia from wastewater in a concentrated form, which is key to its exploitation in the circular economy, but the latent heat demand for thermal separation remains a critical barrier to exploitation. In this study, we investigate the vapor-liquid equilibrium (VLE) for ammonia-water as a mechanism to enhance recovered ammonia quality and minimise the thermal energy required for ammonia separation. Below the dew point (65 °C at 0.25 bar) a two-phase region of the VLE exists where 48 %wt gas-phase ammonia could be produced (61 °C) compared to only 2 %wt within the stripping region adopted widely in the literature. This was complemented by a 98 % reduction in thermal separation energy, since limited water vaporization can occur when the feed is maintained below the activation energy threshold for bulk evaporation. Operation within this practically unexplored region of the ammonia-water VLE fosters a gas-phase product suitable for energy generation in gas turbines or solid oxide fuel cells. Comparable product quality was achieved using concentrated wastewater, which validated the VLE for design in the presence of a broad range of dissolved gases and volatile inorganic compounds. Rapid desorption of CO 2 occurred during vacuum stripping, subsequently increasing pH >9 without the requirement for alkali addition to shift the ammonia-ammonium equilibrium in favor of gaseous ammonia. Consequently, the two-phase region of the VLE defined for vacuum thermal stripping provides a synergistic strategy to mitigate chemical demand, minimise separation energy and recover gas-phase ammonia for zero carbon energy generation, constituting a significant advancement toward the net zero ambitions of the water sector., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2023 The Author(s). Published by Elsevier Ltd.. All rights reserved.)

Existing conventional biological treatment techniques face numerous limitations in effectively removing total petroleum hydrocarbons (TPHs) and ammonia (NH 4 -N) from oilfield-produced water (OPW), highlighting the pressing need for innovative pre-oxidation and biological treatment processes. In this study, a pyrite-activated peroxymonosulfate (PMS)-coupled heterotrophic ammonia assimilation (HAA) system was established to achieve satisfactory system performance for OPW treatment. Pyrite sustained-release Fe + -N) from oilfield-produced water (OPW), highlighting the pressing need for innovative pre-oxidation and biological treatment processes. In this study, a pyrite-activated peroxymonosulfate (PMS)-coupled heterotrophic ammonia assimilation (HAA) system was established to achieve satisfactory system performance for OPW treatment. Pyrite sustained-release Fe 2+ -activated PMS was used to produce SO 4 -N removal efficiencies in the test group with pre-oxidation were 96.9 and 98.3 %, compared to 46.5 and 77.1 % in the control group, respectively. The maximum fluorescence intensities of tryptophan protein and aromatic protein in the test group declined by 83.7 %. Fourier transform ion cyclotron resonance mass spectrometry revealed that pre-oxidation degraded more long-chain hydrocarbons and aromatic family compound, whereas the HAA process produced more proteins and carbohydrates. Pyrite-PMS promoted the enrichment of ammonia-assimilating bacteria, alleviating the explosive increase in extracellular polymeric substances and reducing sludge settleability. The low cost, efficiency, green chemistry principles, and synergies of this approach make it a powerful solution for practical OPW treatment to reduce environmental impacts and promote sustainable wastewater treatment.•- and •OH, and 71.0 % of TPHs were effectively removed from the oil wastewater. The average TPHs and NH 4 + -N removal efficiencies in the test group with pre-oxidation were 96.9 and 98.3 %, compared to 46.5 and 77.1 % in the control group, respectively. The maximum fluorescence intensities of tryptophan protein and aromatic protein in the test group declined by 83.7 %. Fourier transform ion cyclotron resonance mass spectrometry revealed that pre-oxidation degraded more long-chain hydrocarbons and aromatic family compound, whereas the HAA process produced more proteins and carbohydrates. Pyrite-PMS promoted the enrichment of ammonia-assimilating bacteria, alleviating the explosive increase in extracellular polymeric substances and reducing sludge settleability. The low cost, efficiency, green chemistry principles, and synergies of this approach make it a powerful solution for practical OPW treatment to reduce environmental impacts and promote sustainable wastewater treatment., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2023. Published by Elsevier Ltd.)

In this study, a carboxylate platform of hyperthermophilic (70 ℃) anaerobic fermentation (HAF) for short chain fatty acids (SCFAs) production from thermal hydrolyzed sludge (THS) was established. The long-term performance for SCFAs production and the microbial communities of this HAF under different SRTs were systematically investigated. Under the optimum SRT of 3 d, the HAF had the highest acetate production rate of 1.12 g COD/L/d which accounted for 60% in SCFAs. It also rendered a good performance in SCFAs production, with concentration, production rate and yield of 6.61 g COD/L, 1.86 g COD/L/d and 324 g COD/kg VSS in , respectively. Nearly no biogas produced from this system, which reduced the loss of carbon sources from the system. This was due to the inhibition of methanogenesis by the hyperthermophilic condition and the high content of total ammonia nitrogen (TAN) and free ammonia nitrogen (FAN). Tepidimicrobium, Bhargavaea and XBB1006 were the dominant genus-level biomarkers under the optimum SRT, which facilitated the decomposition of monosaccharides, amino acids, terpenoids and polyketides into SCFAs. This work provides an applicable anaerobic carboxylate platform for highly efficient SCFAs production from excess sludge., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2023 Elsevier Ltd. All rights reserved.)

As a stable and effective approach for NO 2 -N accumulation, partial denitrification (PD) could significantly cut down operation cost, and PD/Anammox (PD/A) is a promising nitrogen removal process in wastewater treatment. The biotoxicity of free ammonia (FA) to nitrifying bacteria and anammox bacteria has been demonstrated, while whether FA affects PD bacteria is an open question. Here, long-term operation of PD-fixed bed biofilm reactor (PD-FBBR) treating synthetic wastewater and mature landfill leachate was conducted to reveal the mechanism concerning the effect of FA on PD bacteria. Stable NO - -N accumulation, partial denitrification (PD) could significantly cut down operation cost, and PD/Anammox (PD/A) is a promising nitrogen removal process in wastewater treatment. The biotoxicity of free ammonia (FA) to nitrifying bacteria and anammox bacteria has been demonstrated, while whether FA affects PD bacteria is an open question. Here, long-term operation of PD-fixed bed biofilm reactor (PD-FBBR) treating synthetic wastewater and mature landfill leachate was conducted to reveal the mechanism concerning the effect of FA on PD bacteria. Stable NO 2 - -N accumulation was achieved with NO 3 -N reduction rate improved at low FA concentration (< 12 mg/L), while high FA level (> 25 mg/L) had inhibitory effect on PD bacteria. Under FA stress, more extracellular polymeric substances (EPS) were secreted, and the glnA gene abundance, glutamine synthase concentration, and glutamine concentration in cell and EPS significantly increased, indicating the enhancement of glutamine biosynthesis in PD bacteria for ammonia assimilation played an important role in response to FA stress. Metagenomic sequencing showed that FA stimulated the upregulation of narK (NO - -N to NO 2 - -N transformation ratio (NTR) of 60-70% during 280-day operation with FA ranged from 0 to 20.71 ± 0.23 mg/L, while NTR decreased and maintained at ∼30% when FA reached 40.59 ± 0.19 mg/L. Specific NO x - -N reduction rate improved at low FA concentration (< 12 mg/L), while high FA level (> 25 mg/L) had inhibitory effect on PD bacteria. Under FA stress, more extracellular polymeric substances (EPS) were secreted, and the glnA gene abundance, glutamine synthase concentration, and glutamine concentration in cell and EPS significantly increased, indicating the enhancement of glutamine biosynthesis in PD bacteria for ammonia assimilation played an important role in response to FA stress. Metagenomic sequencing showed that FA stimulated the upregulation of narK (NO 3 - -N/NO 2 - -N antiporter) gene abundance and enhanced uptake of NO 3 - -N and extrusion of NO 2 - -N. Comamonas, unclassified&#95;f&#95;&#95;Comamonadaceae and Thauera were highly enriched in biofilm, which played a key role in the stable NO 2 - -N accumulation. Furthermore, a novel two stage PD/A-FBBR was applied to mature landfill leachate treatment, and satisfactory total inorganic nitrogen removal efficiency ranged from 81.38 ± 3.56% to 89.16 ± 1.57% was obtained at relatively low COD/NO 3 - -N of 2.57-2.84. Overall, these findings demonstrated how PD bacteria respond to FA stress and confirmed the feasibility of PD/A process in high FA wastewater treatment., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2023. Published by Elsevier Ltd.)

Green ammonia production from wastewater via electrochemical nitrate reduction contributes substantially to the realization of carbon neutrality. Nonetheless, the current electrochemical technology is largely limited by the lack of suitable device for efficient and continuous electroreduction nitrate into ammonia and in-situ ammonia recovery. Here, we report a flow-through coupled device composed of a compact electrocatalytic cell for efficient nitrate reduction and a unit to separate the produced ammonia without any pH adjustment and additional energy-input from the circulating nitrate-containing wastewater. Using an efficient and selective Cl-modified Cu foam electrode, nearly 100% NO 3 - electroreduction efficiency and over 82.5% NH 3 Faradaic efficiency was realized for a wide range of nitrate-containing wastewater from 50 to 200 mg NO 3 - -N L -1 . Moreover, this flow-through coupled device can continuingly operate at a large current of 800 mA over 100 h with a sustained NH 3 yield rate of 420 μg h -1 cm -2 for nitrate-containing wastewater treatment (50 mg NO 3 - -N L -1 ). When driven by solar energy, the flow-through coupled device can also exhibit exceptional real wastewater treatment performance, delivering great potential for practical application. This work paves a new avenue for clean energy production and environmental sustainability as well as carbon neutrality., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2023 Elsevier Ltd. All rights reserved.)

Membrane aerated biofilm reactor (MABR) has attracted a lot of attention as an energy-efficient integrated nitrogen removing technology in recent years. However, it is lacking of understanding to realize stable partial nitrification in MABR because of its unique oxygen transfer mode and biofilm structure. In this study, free ammonia (FA) and free nitrous acid (FNA) based control strategies for partial nitrification with low NH 4 + -N concentration were proposed in a MABR of sequencing batch mode. The MABR was operated for over 500 days under different influent NH 4 + -N concentration of around 100 mg-N/L, the FA concentration was lower and strengthened suppression strategies based on FNA were needed. With the final pH of operating cycles below 5.0, the FNA produced in the sequencing batch MABR could stabilize partial nitrification by eliminating NOB on the biofilm. Since the activity of ammonia oxidizing bacteria (AOB) was lower without the blow-off of dissolved carbon dioxide in the bubbleless MABR, longer hydraulic retention time was required to reach a low pH for high concentration of FNA to suppress NOB. After exposures to FNA, the relative abundance of Nitrospira was decreased by 94.6%, while the abundance of Nitrosospira increased greatly which became another dominant AOB genus in addition to Nitrosomonas.4 + -N of around 200 mg-N/L, partial nitrification could be established with relatively low concentration of FA (0.4-2.2 mg-N/L) which suppressed nitrite oxidizing bacteria (NOB) on the biofilm. With lower influent NH 4 + -N concentration of around 100 mg-N/L, the FA concentration was lower and strengthened suppression strategies based on FNA were needed. With the final pH of operating cycles below 5.0, the FNA produced in the sequencing batch MABR could stabilize partial nitrification by eliminating NOB on the biofilm. Since the activity of ammonia oxidizing bacteria (AOB) was lower without the blow-off of dissolved carbon dioxide in the bubbleless MABR, longer hydraulic retention time was required to reach a low pH for high concentration of FNA to suppress NOB. After exposures to FNA, the relative abundance of Nitrospira was decreased by 94.6%, while the abundance of Nitrosospira increased greatly which became another dominant AOB genus in addition to Nitrosomonas., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2023. Published by Elsevier Ltd.)