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This study innovated constructed an activated carbon-loaded nano-zero-valent iron (nZVI-C) enhanced membrane aerated biofilm reactor (MABR) coupled partial nitritation/anammox (PN/A) system for optimizing nitrogen and antibiotics removal. Results showed that nitrogen and antibiotic removal efficiencies of 88.45 ± 0.14% and 89.90 ± 3.07% were obtained by nZVI-C, respectively. nZVI-C hastened Nitrosomonas enrichment (relative abundance raised from 2.85% to 12.28%) by increasing tryptophan content in EPS. Furthermore, nZVI-C proliferated amo gene by 3.92 times and directly generated electrons, stimulating Ammonia monooxygenase (AMO) co-metabolism activity. Concurrently, via antibiotic resistance genes (ARGs) horizontal transfer, Nitrosomonas synergized with Arenimonas and Comamonadaceae for efficient antibiotic removal. Moreover, nZVI-C mitigated antibiotics inhibition of electron transfer by proliferating genes for PN and anammox electron production (hao, hdh) and utilization (amo, hzs, nir). That facilitated electron transfer and synergistic substrate conversion between ammonia oxidizing bacteria (AOB) and anaerobic ammonia oxidizing bacteria (AnAOB). Finally, the high nitrogen removal efficiency of the MABR-PN/A system was achieved., Competing Interests: Declaration of Competing Interest I acknowledge that I have consulted the Guide for Authors and have prepared the manuscript in compliance with the Publishing Policy as described in the Guide for Authors. This manuscript has not been published or presented elsewhere in part or in entirety and is not under consideration by another journal. This study is the original work of the authors, and all authors mutually agree that it should be submitted to your journal. We have read and understood your journal’s policies, and we believe that neither the manuscript nor the study violates any of these. There are no conflicts of interest to declare., (Copyright © 2024 Elsevier B.V. All rights reserved.)

The feasibility of a synergistic endogenous partial denitrification-phosphorus removal coupled anammox (SEPD-PR/A) system was investigated in a modified anaerobic baffled reactor (mABR) for synchronous carbon, nitrogen, and phosphorus removal. The mABR comprising four identical compartments (i.e., C1-C4) was inoculated with precultured denitrifying glycogen-accumulating organisms (DGAOs), denitrifying polyphosphate-accumulating organisms, and anammox bacteria. After 136 days of operation, the chemical oxygen demand (COD), total nitrogen, and phosphorus removal efficiencies reached 88.6 ± 1.0, 97.2 ± 1.5, and 89.1 ± 4.2%, respectively. Network-based analysis revealed that the biofilmed community demonstrated stable nutrient removal performance under oligotrophic conditions in C4. The metagenome-assembled genomes (MAGs) such as MAG106, MAG127, MAG52, and MAG37 annotated as denitrifying phosphorus-accumulating organisms (DPAOs) and MAG146 as a DGAO were dominated in C1 and C2 and contributed to 89.2% of COD consumption. MAG54 and MAG16 annotated as Candidatus_Brocadia (total relative abundance of 16.5% in C3 and 4.3% in C4) were responsible for 74.4% of the total nitrogen removal through the anammox-mediated pathway. Functional gene analysis based on metagenomic sequencing confirmed that different compartments of the mABR were capable of performing distinct functions with specific advantageous microbial groups, facilitating targeted nutrient removal. Additionally, under oligotrophic conditions, the activity of the anammox bacteria-related genes of hzs was higher compared to that of hdh . Thus, an innovative method for the treatment of low-strength municipal and nitrate-containing wastewaters without aeration was presented, mediated by an anammox process with less land area and excellent quality effluent.

The addition of nano zero-valent iron (nZVI) has been proven to improve the efficiency of the anammox process, however, the mechnism is not clear. Here, the effect of nZVI on anammox microbial community was studied by metagenomic sequencing methods. It was found that 50 mg/L nZVI indeed promoted the removal of NH 4 + and NO 2 - of the anammox reactor and significantly improved the relative abundance of AnAOB (Ca. Brocadia) from 42.1% to 52.5%. What's more, 50 mg/L nZVI increased the abundance of c-di-GMP synthesized protein from 148 rpmr to 252 rpmr in the microbial community and decreased the abundance of c-di-GMP degradation protein from 238 rpmr to 204 rpmr, which indirectly led to the enrichment of c-di-GMP in the microbial community. The enrichment of c-di-GMP reduced the motility of microorganisms in the reactor and promoted the secretion of extracellular polymers by bacteria, which is beneficial to the formation of sludge particles in the anammox reactor. In conclusion, this research clarified the mechanism of nZVI promoting the anammox process and provided theoretical guidance for the engineering application of anammox., (Copyright © 2021 Elsevier Ltd. All rights reserved.)

Long-term exposure of anammox process to 1,4-dioxane was investigated using periodic anammox baffled reactor (PABR) under different 1,4-dioxane concentrations. The results generally indicated that PABR (composed of 4 compartments) has robust resistance to 10 mg-dioxane/L. The 1 st compartment acted as a shield to protect subsequent compartments from 1,4-dioxane toxicity through secretion of high extracellular polymeric substance (EPS) of 152.9 mg/gVSS at 10 mg-dioxane/L. However, increasing 1,4-dioxane to 50 mg/L significantly inhibited anammox bacteria; e.g., ~ 93% of total nitrogen removal was lost within 14 days. The inhibition of anammox process at this dosage was most likely due to bacterial cell lysis, resulting in the decrease of EPS secretion and specific anammox activity (SAA) to 105.9 mg/gVSS and 0.04 mg N/gVSS/h, respectively, in the 1 st compartment. However, anammox bacteria were successfully self-recovered within 41 days after the cease of 1,4-dioxane exposure. The identification of microbial compositions further emphasized the negative impacts of 1,4-dioxane on abundance of C. Brocadia among samples. Furthermore, the development of genus Planococcus in the 1 st compartment, where removal of 1,4-dioxane was consistently observed, highlights its potential role as anoxic 1,4-dioxane degrader. Overall, long-term exposure to 1,4-dioxane should be controlled not exceeding 10 mg/L for a successful application., (Copyright © 2021 Elsevier B.V. All rights reserved.)

A distinct shift of bacterial community driven by organic matter (OM) and powder activated carbon (PAC) was discovered in the simultaneous anammox and denitrification (SAD) process which was operated in an anti-fouling submerged anaerobic membrane bio-reactor. Based on anammox performance, optimal OM dose (50 mg/L) was advised to start up SAD process successfully. The results of qPCR and high throughput sequencing analysis indicated that OM played a key role in microbial community evolutions, impelling denitrifiers to challenge anammox's dominance. The addition of PAC not only mitigated the membrane fouling, but also stimulated the enrichment of denitrifiers, accounting for the predominant phylum changing from Planctomycetes to Proteobacteria in SAD process. Functional genes forecasts based on KEGG database and COG database showed that the expressions of full denitrification functional genes were highly promoted in R C , which demonstrated the enhanced full denitrification pathway driven by OM and PAC under low COD/N value (0.11)., (Copyright © 2017 Elsevier Ltd. All rights reserved.)

Membrane bioreactor with non-woven fabric membranes (NWMBR) is developing into a suitable method for anaerobic ammonium oxidation (anammox). As a carrier, non-woven fabric membrane divided total biomass into biofilm and suspended flocs gradually. Total nitrogen removal efficiency was maintained around 82.6% under nitrogen loading rate of 567.4mgN/L/d after 260days operation. Second-order substrate removal and Stover-Kincannon models were successfully used to simulate the nitrogen removal performance in NWMBR. High-throughput sequence was employed to elucidate the underlying microbial community dynamics. Candidatus Brocadia, Kuenenia, Jettenia were detected to affirm the dominant status of anammox microorganisms and 98.2% of anammox microorganisms distributed in biofilm. In addition, abundances of functional genes (hzs, nirK) in biofilm and suspended flocs were assessed by quantitative PCR to further investigate the coexistence of anammox and other microorganisms. Potential nitrogen removal pathways were established according to relevant nitrogen removal performance and microbial community., (Copyright © 2017 Elsevier Ltd. All rights reserved.)

Reducing activity of commensal bacteria in inocula may enhance anammox bacteria proliferation and realization of anammox process. Fast start-up of anammox process in an UASB reactor was successfully achieved by using autoclaved sludge (anaerobic granular sludge pretreated by autoclaving) and 0.3% active anammox sludge as inoculum. Continuous experiments indicated that R2 (autoclaved sludge addition) could shorten the start-up period from 72days to 63days. The first 50days anammox population specific growth rates (μ) of R1 (the control) and R2 were determined to be 0.014d -1 and 0.045d -1 using q-PCR assays. Analysis of coefficient of variations of nitrogen removal performance during days 96-225 indicated that R2 was more stable than R1. The Illumina MiSeq sequencing showed that autoclaving could decrease microbial diversity of sludge and enhance the abundance of anammox bacteria. Furthermore, PICRUSt community functions forecast and c-di-GMP measure illuminated the result of higher stability in R2., (Copyright © 2017 Elsevier Ltd. All rights reserved.)

A permanent microbial stratified nitrogen removal system coupling anammox with partial nitrification (SNAP) in a sequencing batch biofilter granular reactor (SBBGR) was successfully constructed for the treatment of ammonia-rich wastewater. With a nitrogen loading rate of 0.1kgNm -3 ·d -1 , the maximal ammonia and total nitrogen removal efficiencies could reach up to 96.08% and 84.86% on day 108, respectively. The pH, DO profiles revealed a switch of functional species (AOB and anammox) at a typical intermittent aeration cycle. qPCR and high throughput analyses certified a stable spatial microbial stratified community structure. Although, anammox preferred strict anaerobic environment while AOB needed oxygen, a special stratified community structure contributed to conquer this obstacle. Moreover, Bacteroidet, Chlorobi, OD1, Planctomycetes, and Proteobacteria were the dominant species in the SBBGR. Although we have predicted the possible pathways of nitrogen transformation, further studies are needed to validate the pathways in enzymology., (Copyright © 2017 Elsevier Ltd. All rights reserved.)

Partial nitrification (PN) combined with anaerobic ammonium oxidation process has been recognized as a promising technology for the removal of nitrogenous contaminants from wastewater. This research aimed to investigate the potential of external magnetic field for enhancing the PN process in short and long term laboratory-scale experiments. Different strength magnetic fields (0, 5, 10, 15, 20 and 25 mT) were evaluated in short-term batch tests and 5 mT magnetic field was found to have better ability to increase the activities of aerobic ammonium oxidizing bacteria (AOB) of PN consortium. Long-term effect of magnetic field on PN consortium was studied with 5 mT magnetic field. The results demonstrated that the positive effect of magnetic field on PN process could also be testified at all of the four stages. Furthermore, a decrease of bacterial diversity was noted with the increase of magnetic field strength. Relative abundance of Nitrosomonadaceae decreased significantly (p < 0.01) from 13.9% in R CK to 12.9% in R 5mT and 5.5% in R 25mT . Functional genes forecast based on KEGG database indicated that the expressions of functional genes related to signal transduction and cell motility in 5 mT environment were higher expressed compared with no magnetic field addition and high magnetic field addition. The existence of 5 mT magnetic field didn't increase the abundance of AOB but increased the activity of AOB by increasing the rate of free ammonia into the interior of microbial cells. Addition of magnetic field couldn't change the final state of PN process according to the hypothesis proposed in this article. These findings indicated that the weak magnetic field was useful and reliable for the fast start-up of PN process since it was proved as a simple and convenient approach to enhance AOB activity., (Copyright © 2017 Elsevier Ltd. All rights reserved.)

A novel granulation strategy by introducing magnetic nanoparticles (MNPs) into activated sludge system was investigated in this study. The study of the physicochemical characteristics (appearances, sizes, sludge volume index, and chemical oxygen demand) demonstrated that MNPs could decrease the granulation time and improve the retention of biomass, meanwhile enhanced the compact structure of the granules. The secretion and functional groups especially OH and CO of extracellular polymeric substances (EPS) also had significant changes under the long-term influence of MNPs. The contents of proteins (PN) and polysaccharides (PS) in R2 (with MNPs) were 95.7523mg/gVSS and 43.7129mg/gVSS, while in R1 (without MNPs) they were 85.7523mg/gVSS and 32.8632mg/gVSS, respectively. The contact angles of sludge against water dramatically increased with the increase of MNPs concentration, which means that the addition of MNPs could improve the sludge surface hydrophobicity, playing a positive role in the aggregation process., (Copyright © 2016 Elsevier Ltd. All rights reserved.)

To further expand the application of anammox biotechnology, a novel zero-valent iron-assembled upflow anaerobic sludge bed reactor was employed to strengthen anammox performance under low temperature and shock load. Packed with sponge iron and polyester sponge, this novel reactor could speed up the recovery of anammox activity in 12 days and improve the adaptability of anammox bacteria at the temperature of 10-15 °C. The high nitrogen loading rate of 1109.2 mg N/L/day could be adapted in 27 days and the new nitrogen pathway under the effect of sponge iron was clarified by batch experiment. Moreover, the real-time quantitative PCR analysis and Illumina MiSeq sequencing verified the dominant status of Candidatus Kuenenia stuttgartiensis and planctomycete KSU-1, as well as demonstrated the positive role of sponge iron on anammox microorganisms' proliferation. The findings might be beneficial to popularize anammox-related processes in municipal and industrial wastewater engineering.

The long start-up time of anaerobic ammonium oxidation (anammox) process hinders the widespread application of anammox technology in practical wastewater treatment when anammox seed sludge is not available. Meanwhile, the production of nitrate cannot meet the increasingly more strict discharge standards. To combine the chemical nitrate reduction to ammonium with biological nitrogen removal, two anammox upflow anaerobic sludge blanket reactors packed with different types of zero-valent iron (ZVI), microscale ZVI (mZVI) and nanoscale ZVI (nZVI), were developed to accelerate the start-up of anammox process. The results revealed that anammox start-up time shortened from 126 to 105 and 84 days with the addition of mZVI and nZVI. The nitrogen removal performance was also improved remarkably by adding ZVI, especially in the start-up stage. The value of dissolved oxygen showed that ZVI could be regarded as a useful deoxidant to create anaerobic condition for the proliferation of anammox bacteria. ZVI was favorable for the secretion of EPS, which would represent the activity of anammox bacteria. The result of real-time quantitative PCR (qPCR) further confirmed that the proliferation of anammox bacteria was enhanced by ZVI.

From discovery in the early 1990s to completion of full-scale anammox reactor, it took almost two decades to uncover the secret veil of anammox bacteria. There were three milestones during the commercialization of anammox: the development of the first enrichment culture medium, the completion of the first commercial anammox reactor, and the fast start-up of full-scale anammox plant. Till now, the culture of anammox bacteria experienced a big progress through two general strategies: (a) to start up a reactor from scratch and (b) to seed the reactor with enriched anammox sludge. The first full-scale anammox reactor took 3.5 years to realize full operation using the first approach due to several reasons besides the lack of anammox sludge. On the other hand, the first Asian anammox reactor started up in two months, thanks to the availability of anammox seed. Along with the implementation of anammox plants, anammox eventually becomes the priority choice for ammonium wastewater treatment.

The possibility to introduce the exotic anammox sludge to seed the pilot-scale anammox granular reactor and its fast start-up for treating high nitrogen concentration wastewater were evaluated in this study. The reactor was started up successfully in two weeks; in addition, high nitrogen removal was achieved for a long period. Stoichiometry molar ratios of nitrite conversion and nitrate production to ammonium conversion were calculated to be 1.26±0.02:1 and 0.26±0.01:1, respectively. The Stover-Kincannon model which was first applied in granular anammox process indicated that the granular anammox reactor possessed high nitrogen removal potential of 27.8 kg/m(3)/d. The anammox granules in the reactor were characterized via microscope observation and fluorescence in situ hybridization technique. Moreover, the microbial community of the granules was quantified to be composed of 91.4-92.4% anammox bacteria by real-time polymerase chain reaction. This pilot study can elucidate further information for industrial granular anammox application., (Copyright © 2010 Elsevier Ltd. All rights reserved.)

Granular anammox reactors usually adopted anaerobic/aerobic granules as source sludge, in which the washout of other species and enrichment of anammox biomass were very slow because of the competition of the coexisting bacteria. In this study, inactive methanogenic granules were proved to be suitable for rapid anammox granulation under high nitrogen concentrations by investigating their interaction with anammox bacteria. The start-up nitrite concentration was significantly higher than the published toxic level for anammox bacteria and other lab-scale studies. The nitrogen loading rate increased from 141 to 480 mg/L/d in 120 days operation with a total nitrogen removal efficiency of 96.0+/-0.6%. Anammox granules with a diameter of 1.3+/-0.4mm were observed over the course of three months. Molecular analysis showed that over 67% of the cells in the anammox granules were anammox bacteria after 90 days. The accommodations and proliferations of anammox bacteria in the inactive methanogenic granules might be the main reason for the high anammox purity in a short period. The important role of the extracellular polymer in the granule structure was observed via morphological observation., (Copyright 2010 Elsevier Ltd. All rights reserved.)

The anammox non-woven membrane reactor (ANMR) is a novel reactor configuration to culture the slowly growing anammox bacteria. Different mathematical models were used to study the process kinetics of the nitrogen removal in the ANMR. The kinetics of nitrogen gas production of anammox process was first evaluated in this paper. For substrate removal kinetics, the modified Stover-Kincannon model and the Grau second-order model were more applicable to the ANMR than the first-order model and the Monod model. For nitrogen gas production kinetics, the Van der Meer and Heertjes model was more appropriate than the modified Stover-Kincannon model. Model evaluation was carried out by comparing experimental data with predicted values calculated from suitable models. Both model kinetics study and model testing showed that the Grau second-order model and the Van der Meer and Heertjes model seemed to be the best models to describe the nitrogen removal and nitrogen gas production in the ANMR, respectively., ((c) 2010 Elsevier Ltd. All rights reserved.)

An innovative reactor configuration for Anammox enrichment by connecting a non-woven membrane module with an anaerobic reactor was developed in this study. The Anammox non-woven membrane reactor (ANMR) exhibited high biomass retention ability through the formation of aggregates in the reactor and biofilm on the interior surface of the non-woven membrane. No fouling problems occurred on the membrane after the development of mature biofilms. After 8 months of operation, the nitrogen loading rate (NLR) and nitrogen removal rate (NRR) reached 1263 mg N/l/d and 1047.5 mg N/l/d, respectively, with a maximum specific ammonium consumption (SAC) of 51 nmol/mg protein/min. At steady state, the average ammonium and nitrite removal efficiencies were 90.9% and 95.0%, respectively. Morphological observation of Anammox aggregates and biofilm showed a high degree of compactness. Also, enrichment of Anammox bacteria was quantified by real-time polymerase chain reaction (PCR) analysis as 97.7%., (Copyright (c) 2009 Elsevier Ltd. All rights reserved.)

Dissimilatory nitrate reduction to ammonium (DNRA) received more attention for its ability to recover ammonium. This study investigated the possibility of low-frequency infrared electromagnetic field (IR-EMF) to improve DNRA. The optimal IR-EMF intensity of 0.04 μT could effectively improve DNRA activity of nonwoven fabric membrane bioreactors. In the long-term operation, the average ammonium conversion efficiency was enhanced by 117.7% and 62.5% under 0.04 μT and 0.06 μT IR-EMF, respectively. The highest nrfA-gene abundance and potential DNRA rate were obtained under 0.04 μT IR-EMF exposure. Bacteroidetes fragilis, Shewanelle oneidensis MR-1, and Thauera sp. RT1901 were selected to investigate the dynamic response of nitrogen transformation and energy metabolism to IR-EMF. The transcriptome sequencing and RT-qPCR results suggested that IR-EMF could enhance both denitrification and DNRA process, mainly by improving ATP synthesis to boost metabolic activity. This study provided an efficient method for the nitrogen recovery via DNRA process by applying IR-EMF. [ABSTRACT FROM AUTHOR]

Extracellular matrix plays a significant role in formation of matrix structure, biogranulation process and improvement of stability of anammox granules. Distributions of cells and extracellular polymeric substances (EPS) in anammox granules cultured from activated sludge and inactive methanogenic granules were probed. Anammox bacteria secreted more EPS than anaerobic/aerobic granules. The layer of EPS surrounding anammox cells was thicker than other types of granules. In high-enriched granules, the proteins and β- d -glucopyranose polysaccharides were principally distributed at the core, whereas the cells and α- d -glucopyranose polysaccharide accumulated in both the interior and outer layers of granules. In low-enriched ones, cells and α- d -glucopyranose polysaccharides were mainly distributed at the outer rim and the β- d -glucopyranose polysaccharides were located in both the core and the outer layer, whilst the proteins were distributed throughout the whole structure of granules. EPS distribution indicated that low-enriched granules possessed higher stability than high-enriched ones, in consistent with granule strength test. [ABSTRACT FROM AUTHOR]

Membrane bioreactor with non-woven fabric membranes (NWMBR) is developing into a suitable method for anaerobic ammonium oxidation (anammox). As a carrier, non-woven fabric membrane divided total biomass into biofilm and suspended flocs gradually. Total nitrogen removal efficiency was maintained around 82.6% under nitrogen loading rate of 567.4 mg N/L/d after 260 days operation. Second-order substrate removal and Stover-Kincannon models were successfully used to simulate the nitrogen removal performance in NWMBR. High-throughput sequence was employed to elucidate the underlying microbial community dynamics. Candidatus Brocadia , Kuenenia , Jettenia were detected to affirm the dominant status of anammox microorganisms and 98.2% of anammox microorganisms distributed in biofilm. In addition, abundances of functional genes ( hzs , nirK ) in biofilm and suspended flocs were assessed by quantitative PCR to further investigate the coexistence of anammox and other microorganisms. Potential nitrogen removal pathways were established according to relevant nitrogen removal performance and microbial community. [ABSTRACT FROM AUTHOR]