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In this study, a high-rate CANON (Complete Autotrophic Nitrogen-removal Over Nitrite) process was started up successfully by enhancing the in-situ enrichment of anammox bacteria in aerobic granules at conditions relevant for mainstream wastewater treatment. Firstly, to provide nitrite for anammox bacteria growth efficient nitrite-oxidizing bacteria (NOB) repression was rapidly achieved and stably maintained. Both low dissolved oxygen (DO) and ammonium concentrations ratio (DO/NH 4 + <0.15) and selective washing-out of NOB-preferred smaller particles at short hydraulic retention time (HRT, 25-15 min) contributed to the NOB repression. Then the stepwise down-regulating DO concentrations from 2.8 to 1.2 mg/L enhanced the enrichment of anammox bacteria in the aerobic granules. The enriched anammox species was dominated by Ca. Brocadia sapporoensis with the estimated growth rate of 0.008-0.013 d -1  at 15 °C. Chloroflexi and Chlorobi-affiliated bacteria were also significantly enriched in the granules, which may benefit the anammox bacteria activity and growth. At the end of this study, the average total nitrogen removal rate and efficiency of the granular CANON process respectively reached 1.26 kg N·m -3 ·d -1 and 68% treating low-strength ammonium (∼50 mg N·L -1 ) wastewater under such aggressive conditions (DO = 0.8-1.5 mg/L, HRT< 1.0 h, and T = 15 °C). Overall, the aerobic granules provided a habitable niche for the proliferation and almost complete retention of the anammox bacteria. This study provides a roadmap for in-situ starting up of high-rate CANON process for mainstream wastewater treatment with aerobic granules as inoculum., (Copyright © 2021 Elsevier Ltd. All rights reserved.)

To elucidate how high dissolved oxygen (DO) favors the startup of nitritation with aerobic granular sludge, two granular reactors were operated under low (1-2 mg O 2 ·L -1 ) and high DO (3-5 mg O 2 ·L -1 ) conditions with similar effluent ammonium concentrations (>20 mg N·L -1 ). The results showed that though nitritation with an average nitrite accumulation ratio of above 95% was finally achieved in both reactors, a five-fold start-up time (eleven weeks) was required for the low DO reactor compared to the high DO reactor. Moreover, the nitritation performance was positively correlated with the extent of nitrifiers stratification in granules. The faster startup of nitritation under high DO conditions mainly resulted from the faster formation of well-stratified nitrifiers, with ammonium oxidizing bacteria (AOB) dominating granule surface. High DO operation combined with sufficient ammonium supply ensured the faster growth of AOB, which should provide a competitive advantage to AOB in competing for habitable space (i.e., granule surface). Besides, the lower porosity, larger size, and more active extracellular polymeric substances (particularly proteins) production of granules was observed under the high DO condition. Overall, these findings supported the proposition that the switch from mixed to stratified distribution of nitrifiers in granule was primarily driven by their competition for habitable space rather than by oxygen-limitation., 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 © 2020 Elsevier Ltd. All rights reserved.)

Biomass segregation between granules/biofilm and flocs is widespread in anammox-based processes. The segregation of biomass allows for easy control of processes stability. The goal of this study is to understand the biomass segregation in two anoxic anammox reactors respectively operated in nitrite-limited (R NO2 ) and ammonium-limited (R NH4 ) modes treating low-strength wastewater at 20 °C. Results showed that size-based biomass segregation was developed in both reactors. But the functional and population heterogeneity was more significant in the ammonium-limited anammox reactor. The activity and abundance of anammox bacteria in large granules were significantly higher than that in flocs under the ammonium-limited conditions. The large granules played a major role in nitrogen removal in R NH4 . By contrast, both large granules and small flocs contributed significantly to the nitrogen loss in the nitrite-limited anammox reactor, since a large number of anammox bacteria existed in both granules and flocs. Besides, a number of Nitrospira-like NOB were also detected in both anoxic anammox reactors, which primarily inhabited in flocs seemingly droved by the availability of oxygen. But the abundance of Nitrospira in R NH4 was much higher than that in R NO2 . All these results suggested that selective flocs removal would be necessary for R NH4 to improve its anammox performance but non-essential for R NO2 . The two anammox reactors shared the predominant anammox species with the closest relative to Ca. Brocadia sp. 40 (98%). Unexpectedly, the anammox species grew faster in R NH4 . But the microbial diversity and evenness was much greater in R NO2 , suggesting its higher functional stability., 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 © 2020 Elsevier Ltd. All rights reserved.)

Anammox process was regarded to be one of the vital links to achieve energy-saving or energy-producing wastewater treatment plant. In the study, an anammox reactor with the nitrogen gas circulation was constructed to culture anammox granules, and the performance, granule size distribution, and microbial community were investigated. Dissolved oxygen loading is found to be an important factor for the start-up of the anammox process, and the nitrogen removal rate of 2.12 kg N m -3  day -1 was achieved under the average nitrogen loading rate of 2.6 kg N m -3  day -1 . The activity test showed that the highest specific anammox activity of 345.9 mg N gVSS -1  day -1 was achieved for granules with size of 0.5-1.0 mm. The Illumina high-throughput sequencing analysis revealed the consistent variation of Candidatus Brocadia and Denitratisoma abundance in granues of all sizes, suggesting possible synergistic mechanism between heterotrophic bacteria Denitratisoma and anammox bacteria Ca. Brocadia. Furthermore, the results indicated the reactor with the nitrogen gas circulation is an efficient strategy to start-up anammox.

The effects of influent shift from synthetic wastewater to anaerobically pretreated actual sewage coupling with lowering temperature on microbial community of a two-stage partial nitritation (PN)-anammox process were evaluated through high-throughput sequencing. Venn diagrams and Hill numbers showed the significantly increased bacterial diversity both in the PN and anammox reactor. However, taxonomic analysis indicated that outstanding enrichment of heterotrophic bacteria and reduction of autotrophic species mainly occurred in the PN reactor, while nearly all of the dominant bacteria in the anammox reactor only slightly decreased in abundance. Moreover, immigrant bacteria from the PN reactor to the following anammox reactor had no negative effect on the anammox function. These results implied the positive role of the first-stage PN in maintaining the stability of the following anammox community. Nitrosomonas europaea (17.9-52.9%) and one cluster (19.2-27.7%) within Candidatus Brocadia remained as the dominant functional species in the PN and anammox reactor, respectively., (Copyright © 2018 Elsevier Ltd. All rights reserved.)

The partial nitrification (PN) performance and the microbial community variations were evaluated in a sequencing batch reactor (SBR) for 172 days, with the stepwise elevation of ammonium concentration. Free ammonia (FA) and low dissolved oxygen inhibition of nitrite-oxidized bacteria (NOB) were used to achieve nitritation in the SBR. During the 172 days operation, the nitrogen loading rate of the SBR was finally raised to 3.6 kg N/m 3 /d corresponding the influent ammonium of 1500 mg/L, with the ammonium removal efficiency and nitrite accumulation rate were 94.12% and 83.54%, respectively, indicating that the syntrophic inhibition of FA and low dissolved oxygen contributed substantially to the stable nitrite accumulation. The results of the 16S rRNA high-throughput sequencing revealed that Nitrospira, the only nitrite-oxidizing bacteria in the system, were successively inhibited and eliminated, and the SBR reactor was dominated finally by Nitrosomonas, the ammonium-oxidizing bacteria, which had a relative abundance of 83%, indicating that the Nitrosomonas played the primary roles on the establishment and maintaining of nitritation. Followed by Nitrosomonas, Anaerolineae (7.02%) and Saprospira (1.86%) were the other mainly genera in the biomass.

The effect of pH in the range of 3.0-11.0 on anaerobic fermentation of primary sludge (PS) was investigated at room temperature. The experimental results showed that the concentrations of soluble chemical oxygen demands (SCOD), soluble protein and carbohydrate and short-chain fatty acids (SCFAs) under alkaline conditions were significantly higher than those under other pHs. At fermentation time of 5 days, the average SCFAs concentration increased from 968 to 3511mg COD/L with the increase of pH from 3.0 to 10.0. However, further increasing pH to 11.0 resulted in the decrease of SCFAs. At pH 10.0, acetic, propionic and iso-valeric acids were the three main products, and the volatile suspended solids (VSS) reduction reached 38%. It was also observed that at any pH value investigated, there were obvious ammonia and phosphorus releases during fermentation. According to this study it is obvious that alkaline pH benefited the soluble organic carbon production from PS.

Abstract: A new pathway was explored in order to gain insights into the role of biopolymers in sludge characteristics and membrane fouling. The biopolymers were discharged from a membrane bioreactor (MBR) using a unique design (called as dual-MBR in this study). The variations of soluble microbial products (SMP), extracellular polymeric substances (EPS), sludge surface charge, particle size and dewaterability were analyzed in the dual-MBR after the discharging of biopolymers from the system. It was found that carbohydrates in SMP, proteins in SMP and humics in SMP were decreased in the dual-MBR compared to the control-MBR. Gel filtration chromatography (GFC) analysis showed that the substances with macromolecular weight in SMP were reduced in the dual-MBR. Three-dimensional excitation–emission matrix (EEM) fluorescence spectra demonstrated that the aromatic protein-like substances in SMP were also lowered in the dual-MBR. The neutral hydrophilic (HPI-N) fraction in SMP, which was found to have larger molecular weight distribution and stronger fouling potential compared to the rest fractions, was significantly reduced in the dual-MBR. The exclusion of biopolymers from the dual-MBR also resulted in the decrease of the bound EPS and the reduction of proteins in bound EPS, which consequently caused the decrease of sludge surface charge, the increase of sludge flocs and the improvement of sludge dewaterability. The above changes induced by the exclusion of biopolymers can explain why the trans-membrane pressure in the dual-MBR exhibited a slower increase rate compared to the control-MBR. This study offered an evaluation of the contribution of biopolymers to sludge characteristics and membrane fouling through a different angle from previous publications. [Copyright &y& Elsevier]

Abstract: Four flat-sheet membrane modules, which were operated under four different filtration modes but with the same treatment capacity, were used to treat synthetic wastewater in a submerged membrane bioreactor (MBR). Particle size distribution (PSD), gel filtration chromatography (GFC), capillary suction time (CST), and three-dimensional excitation–emission matrix (EEM) fluorescence spectroscopy were used to characterize membrane fouling properties. The high instantaneous flux induced faster fouling rate and continuous filtration mode was the most applicable filtration mode in this study. The average particle size of all foulants was smaller than that of bulk sludge; and the higher the instantaneous flux was adopted, the larger the average particle size of foulants would be. Only macromolecule substances were detected in all the foulants. The macromolecule substances in the influent were degraded by microorganism and retained by membrane, and small molecular substances could pass through membrane pores to enter the effluent. The membrane foulants had poorer dewaterability compared to the mixed liquor confirmed by CST measurement. Although there were several peaks associated with protein-like fluorophores, fulvic acid-like substances and humic acid-like organics in soluble microbial products (SMP) and extracellular polymeric substances (EPS) sample, it was found that the dominant fluorescence substances in membrane foulants were protein-like substances. [ABSTRACT FROM AUTHOR]

Abstract: Membrane foulants were systematically characterized in an anaerobic membrane bioreactor (AnMBR), in which non-woven fabric was used as the membrane material. In this study, three-dimensional excitation-emission matrix (EEM) fluorescence spectroscopy, gel filtration chromatography (GFC), Fourier transform infrared (FT-IR) spectroscopy, scanning electron microscopy (SEM) and energy-diffusive X-ray (EDX) analyzer were used to analyze the membrane foulants. The results indicated that the organic substances with fluorescence characteristics in the extracellular polymeric substances (EPS) extracted from the membrane foulants were identified as proteins and visible humic acid-like substances by EEM technology. The GFC analysis exhibited that the EPS had much broader distributions of molecular weight (MW) and a larger weight-average molecular weight (M w) compared with the influent wastewater and the effluent. Proteins and clay materials were also identified in the fouling layer by the FT-IR analysis. The examination by EDX demonstrated that Mg, Al, Ca, Si, and Fe were the major inorganic elements in the fouling cake. Furthermore, the results suggested that bridging between deposited biopolymers and inorganic compounds could enhance the compactness of fouling layer. During the operation of AnMBR, the protein-like and visible humic acid-like substances, which were main components of the organic matters with fluorescence characteristics in the influent, were partly removed. And the large MW organics (>4000Da) in the influent were partly metabolized into low MW organics by microorganisms combined with the separation function of the cake layer and the membrane pores. [Copyright &y& Elsevier]