•Higher external mass transfer resistance favours NOB repression.•IFAS has reduced operating conditions range despite achieving higher removal rates.•AOB seeding from biofilm is crucial for stability ...of IFAS systems.•Anammox nitrite sink is strongly affected by the inhibition of anammox by oxygen.
A model-based study was developed to analyse the behaviour of Moving Bed Biofilm Reactor (MBBR) and Integrated Fixed-Film Activated Sludge (IFAS) reactor configurations for the removal of nitrogen in the main water line of municipal wastewater treatment plants via partial nitritation/anammox (PN/AMX). The basic principles and underlying mechanisms linking operating conditions to process performance were investigated, with particular focus on nitrite oxidizing bacteria (NOB) repression and resulting volumetric conversion rates. The external mass transfer resistance is a major factor differentiating granular sludge PN/AMX processes from MBBR or IFAS systems. The external mass transfer resistance was found to promote the metabolic coupling between anammox (AMX) and ammonia oxidizing bacteria (AOB), crucial for NOB repression in the biofilm. Operation at low bulk DO prevents NOB proliferation in the flocs of IFAS systems as AMX activity limits nitrite availability (the so-called AMX nitrite sink). Importantly, the effectiveness of the AMX nitrite sink strongly depends on the AMX sensitivity to oxygen. Also, over a broad range of operational conditions, the seeding of AOB from the biofilm played a crucial role in maintaining their activity in the flocs. From a practical perspective, while low DO promotes NOB repression, lower nitrogen loads have to be applied to maintain the same effluent quality. Thus, a trade-off between NOB repression and volumetric conversion capacity needs to be defined. To this end, IFAS allow for higher volumetric rates, but the window of operating conditions with effective NOB repression is smaller than that for MBBR. Ultimately, this study identified the principles controlling NOB in MBBR and IFAS systems and the key differences with granular reactors, allowing for the interpretation of (seemingly contradictory) published experimental results.
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Membrane aerated biofilm reactor (MABR) and shortcut nitrogen removal are two types of solutions to reduce energy consumption in wastewater treatment, with the former improving the aeration ...efficiency and the latter reducing the oxygen demand. However, integrating these two solutions, i.e., achieving shortcut nitrogen removal in MABR, is challenging due to the difficulty in suppressing nitrite-oxidizing bacteria (NOB). In this study, four MABRs were established to demonstrate the feasibility of initiating, maintaining, and restoring NOB suppression using low dissolved oxygen (DO) control, in the presence and absence of anammox bacteria, respectively. Long-term results revealed that the strict low DO (< 0.1 mg/L) in MABR could initiate and maintain stable NOB suppression for more than five months with nitrite accumulation ratio above 90 %, but it was unable to re-suppress NOB once they prevailed. Moreover, the presence of anammox bacteria increased the threshold of DO level to maintain NOB suppression in MABRs, but it was still incapable to restore the deteriorated NOB suppression in conjunction with low DO control. Mathematical modelling confirmed the experimental results and further explored the differences of NOB suppression in conventional biofilms and MABR biofilms. Simulation results showed that it is more challenging to maintain stable NOB suppression in MABRs compared to conventional biofilms, regardless of biofilm thickness or influent nitrogen concentration. Kinetic mechanisms for NOB suppression in different types of biofilms were proposed, suggesting that it is difficult to wash out NOB developed in the innermost layer of MABR biofilms because of the high oxygen level and low sludge wasting rate. In summary, this study systematically demonstrated the challenges of NOB suppression in MABRs through both experiments and mathematical modelling. These findings provide valuable insights into the applications of MABRs and call for more studies in developing effective strategies to achieve stable shortcut nitrogen removal in this energy-efficient configuration.
The development of new wastewater treatment processes can assist in reducing the impact of wastewater treatment on the environment. The recently developed partial nitritation anammox (PNA) process, ...for example, consumes less energy for aeration and reduces nitrate in the effluent without requiring additional organic carbon. However, achieving stable nitritation (ammonium oxidation; NH4+ → NO2−) at mainstream conditions (T = 10–25 °C, C:N > 10, influent ammonium < 50 mgNH4-N/L and effluent < 1 mgNH4-N/L) remains challenging. This study explores the potential and mechanism of nitrite-oxidizing bacteria (NOB) suppression in a bottom-fed sequencing batch reactor (SBR). Two bench-scale (11 L) reactors and a pilot-scale reactor (8 m3) were operated for over a year and were fed with organic substrate depleted municipal wastewater. Initially, nitratation (nitrite oxidation; NO2− → NO3−) occurred occasionally until an anaerobic phase was integrated into the operating cycle. The introduction of the anaerobic phase effectively suppressed the regrowth of NOB while nitritation was stable over 300 days, down to 8 °C and at ammonium influent concentrations < 25 mgNH4-N/L. Batch experiments and process data revealed that parameters typically affecting NOB growth (e.g., dissolved oxygen, alkalinity, trace elements, lag-phase after anoxia, free nitrous acid (FNA), free ammonia (FA), pH, sulfide, or solids retention time (SRT)) could not fully explain the suppression of nitratation. Experiments in which fresh nitrifying microbial biomass was added to the nitritation system indicated that NOB inactivation explained NOB suppression better than NOB washout at high SRT. This study concludes that bottom-fed SBRs with anaerobic phases allow for stable nitritation over a broad range of operational parameters. Coupling this type of SBR to an anammox reactor can enable efficient mainstream anammox-based wastewater treatment.
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•Nitritation was stable in municipal wastewater, even at low temperatures (8 °C).•A one-hour anaerobic phase, after reactor filling, rapidly established nitritation.•Bottom-feeding was used for a 90% volume exchange each SBR cycle.•NOB suppression is driven by NOB inactivation rather than washout.•Up-scaling of nitritation to an 8 m3 reactor was successful.
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•An integrated strategy for stabilizing NOB inhibition was successfully established.•Nutrient conversion pathways that PNA coupled with EPDA were specified.•Advanced nitrogen and ...phosphorus removal was achieved in mainstream PNA system.•Functional microbial activities, composition, and variations were investigated.•Anammox-related functional genes had higher expression at the transcriptome level.
Partial nitrification anammox (PNA) is a promising technology for energy saving and nitrogen removal. The bottlenecks limiting the widespread application of PNA in mainstream municipal wastewater treatment include the difficulty in stably suppressing nitrite-oxidizing bacteria (NOB) and failure to meet discharge standards for total nitrogen (TN) due to high nitrate concentration in the effluent. To this end, this study aims to stably suppress NOB using integrated strategies (including the inoculated activated sludge pretreatment with FNA, low DO, residual ammonia nitrogen, and floc sludge discharge) while also investigating the feasibility and performance of coupling PNA with endogenous partial denitrification/anammox (EPDA) in a PNA granular hybrid system. During the 300 days of operation, the integrated strategies effectively stabilized the suppression of NOB, decreasing its abundance from 1.22 % to 0.55 %, and the final activity was reduced to 0.36 ± 0.12 mg-N/L/h. Meanwhile, the relative abundance of AnAOB as the dominant microbial community increased from 9.16 % to 24 %, with its activity reaching 2.84 ± 0.16 mg-N/L/h, limiting the supply of nitrite and ensuring the stable suppression of NOB. Typical cycle tests showed that EPDA and PNA were successfully coupled during the aeration stage with a nitrogen removal contribution rate of 80.55 %, ultimately achieving the average effluent TN and PO43--P were 4.22 mg/L and 0.09 mg/L, respectively. The metagenomic and metatranscriptomic sequencing results revealed that anammox bacteria (AnAOB) coexisted with ammonia-oxidizing bacteria (AOB), NOB, and endogenous denitrifying bacteria (EDB), and AnAOB played a primary role in the nitrogen removal process of mainstream PNA. This study brings mainstream PNA closer to widespread application and provides significant technical support for its engineering implementation.
•PN/AMX pilot plant treated municipal wastewater without temperature control.•Stable two-stage system coping with wastewater characteristic fluctuations.•NOB suppression was rapidly started up by ...overloading and maintained by FNA.•NOB were inactive but still present in the nitritation unit.•Nitrogen removal efficiencies of 80 % were achieved in the anammox reactor.
Two-stage partial nitritation (PN) and anammox (AMX) systems showed promising results for applying autotrophic nitrogen removal under mainstream conditions. In this study, a pilot-scale (600 L per reactor) two-stage PN/AMX system was installed in a municipal wastewater treatment plant (WWTP) provided with a high-rate activated sludge (HRAS) system for organic carbon removal. The PN/AMX system was operated without temperature control (ranging from 11 to 28 °C) and was subjected to the same variations in wastewater characteristics as the WWTP (22 to 63 mg NH4+- N/L). The developed strategy is simple, does not require the addition of chemicals and is characterised by short start-up periods. The PN process was established by applying a high hydraulic load and maintained by in situ accumulated free nitrous acid (FNA) of 0.015–0.2 mg HNO2-N/L. Based on pH value, a controlled aeration strategy was applied to achieve the target nitrite to ammonium ratio in the effluent (1.1 g NO2–-N/g NH4+-N) to feed the AMX reactor. Although NOB were not fully washed out from the system, nitrite accumulation remained (>99 %) stable with no evidence of NOB activity. In the AMX reactor, an overall nitrogen removal efficiency of 80 % was achieved. Regarding effluent quality, 12 ± 3 mg TN/L was obtained, but 5 mg NO3–-N/L was already in the HRAS effluent. The relative abundance of NOB showed a strong negative correlation with the FNA concentration, providing a good strategy for establishing PN under mainstream conditions.
Hepatic lipidome has been given emphasis for years since hepatic steatosis is the most remarkable character of nonalcoholic fatty liver diseases, an increasingly serious health issue worldwide. ...Nobiletin (NOB), one of the citrus flavonoids, exerted outstanding effect on lipid metabolism disorder. However, the underlying mechanism of NOB exerting effect on hepatic lipid alternation remains unclear. In this study, the animal model was built by feeding APOE−/− mice with high fat diet (HFD). The results of Oil Red O-stained liver section and the biochemical assay of lipid parameters confirmed the protective effect of NOB on hepatic steatosis and global lipid metabolism disorder in APOE−/− mice. The hepatic lipidomic study revealed a total of 958 lipids significantly altered by HFD and a total of 86, 116, 212 lipid metabolites changed by L-NOB (50 mg/kg/d NOB), M-NOB (100 mg/kg/d NOB) and H-NOB (200 mg/kg/d NOB) respectively. In the further screening analysis, an amount of 60 lipids were identified as the potential lipid markers of NOB treatment, most of which belonged to glycerophospholipids lipid categories and exhibited obvious correlation with each other and the lipid parameters related to hepatic steatosis. Taken together, our data demonstrated that glycerophospholipids metabolism played an indispensable role in the progression of hepatic steatosis and the protective effect of NOB. Besides, the modulation towards genes involved in lipid synthesis was observed after NOB administration in this study. These finding illustrated the antihepatic steatosis effect of NOB based on altering hepatic lipidome, particularly the glycerophospholipids metabolism, and provided a new insight in the pathogenesis of hepatic steatosis.
Microbial nitrite oxidation is the primary pathway that generates nitrate in wastewater treatment systems and can be performed by a variety of microbes: namely, nitrite-oxidizing bacteria (NOB). ...Since NOB were first isolated 130 years ago, the understanding of the phylogenetical and physiological diversities of NOB has been gradually deepened. In recent endeavors of advanced biological nitrogen removal, NOB have been more considered as a troublesome disruptor, and strategies on NOB suppression often fail in practice after long-term operation due to the growth of specific NOB that are able to adapt to even harsh conditions. In line with a review of the history of currently known NOB genera, a phylogenetic tree is constructed to exhibit a wide range of NOB in different phyla. In addition, the growth behavior and metabolic performance of different NOB strains are summarized. These specific features of various NOB (e.g., high oxygen affinity of Nitrospira, tolerance to chemical inhibitors of Nitrobacter and Candidatus Nitrotoga, and preference to high temperature of Nitrolancea) highlight the differentiation of the NOB ecological niche in biological nitrogen processes and potentially support their adaptation to different suppression strategies (e.g., low dissolved oxygen, chemical treatment, and high temperature). This review implicates the acquired physiological characteristics of NOB to their emergence from a genomic and ecological perspective and emphasizes the importance of understanding physiological characterization and genomic information in future wastewater treatment studies.
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•Mainstream PN was quickly established by sludge treatment using high salinity.•High NAR (95.69 ± 2.65 %) was maintained stable in the mainstream CSTR.•Salinity increased the relative ...abundance of Nitrosomonas from 5.98% to 38.08%.•NOB activity was almost undetectable after sludge treatment using high salinity.•The absolute activity change of NOB should be given attention for sludge treatment.
The difficulty of the start-up and maintenance the stability of partial nitritation (PN) was considered as a key challenge for mainstream anammox. This study demonstrated a novel approach for achieving rapid start-up of mainstream PN by sludge treatment using high salinity based on the phenomenon which high salinity is far more biocidal to nitrite-oxidising bacteria (NOB) than to ammonia-oxidising bacteria (AOB) in short-term (within 24 h). In this study, one-third of the nitrifying sludge from the continuous stirred tank reactor (CSTR) fed by mainstream wastewater was suppressed at 70 g NaCl/L for 24 h every day. The treated nitrifying sludge was afterwards returned back to the CSTR. Mainstream PN with high nitrite accumulation ratio (NAR) was rapidly established (in 14 d) after optimizing the suppression conditions and maintained stable (NAR = 95.69 ± 2.65 %) more than 45 days in the mainstream reactor, indicating the establishment of PN process. After long-term high salinity treatment, NOB activity in the reactor was almost undetectable, while AOB activity significantly increased. Microbial community analysis revealed that the sludge treatment using high salinity could significantly reduce Nitrospira abundance while the relative abundance of Nitrosomonas increased by an order of magnitude. Moreover, the relationship between the absolute activity change of nitrifying sludge and inhibition strength was explored based on the activity batch assays and quantitative polymerase chain reaction assays analysis. The approach of achieving mainstream PN by sludge treatment using high salinity is economically and environmentally attractive because salt is an easily obtainable substance.
•A continuous flow reactor for treating actual municipal wastewater was studied•Nitritation was quickly initiated in 15 d via UL-SRT during seasonal warming•Stable nitritation could be achieved by ...using joint UL-SRT and tapered aeration•The AOB (amoA) abundance increased approximately 50 times after nitritation
Rapid achievement of nitritation of mainstream municipal wastewater in a continuous-flow process is attractive since it favors the involvement of the anammox process and reduces the operational costs. In this study, a feasible and economical strategy is proposed to rapidly achieve the nitritation of municipal wastewater. By aggressively discharging excess sludge during the seasonal warming period (temperature increasing from 18°C to 22°C), nitritation was established in 15 days with a nitrite accumulation ratio of 85.09% in a continuous-flow anaerobic/oxic (An/O) reactor. Meanwhile, qPCR results revealed that amoA abundance increased from (1.78±0.10) × 108 copies/(g VSS) to (1.05±0.11) × 1010 copies/(g VSS) while the abundance of nitrite-oxidizing bacteria decreased from (1.1±0.02) × 1010 copies/(g VSS) to (5.01±0.02) × 108 copies/(g VSS). The temperature gradually stabilized at 26°C during the following operational period and stable nitritation was maintained with a nitrite accumulation ratio above 90%, which was mainly attributed to a short sludge retention time (SRT) of 4.3 days and a low dissolved oxygen of 0.86 ± 0.5 mg/L. Falling temperature negatively impacted the stability of nitritation, but nitritation could be restarted by aggressively discharging excess sludge during another temperature increase period. Overall, this study provides a feasible strategy to start-up nitritation that has great potential applications for municipal wastewater treatment.
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•Inactivation and adaptation of AOB and NOB to FNA was investigated.•FNA caused a stronger inactivation effect on NOB than on AOB.•This inactivation increased as the FNA increased and exposure time ...lengthened.•AOB did not adapt to the FNA treatment. In contrast, NOB did adapt to FNA.•NOB adaptation may be due to its shift from Nitrospira to Candidatus Nitrotoga.
Inactivation and adaptation of ammonia-oxidizing bacteria (AOB) and nitrite-oxidizing bacteria (NOB) to free nitrous acid (FNA) was investigated. Batch test results showed that AOB and NOB were inactivated when treated with FNA. After an 85-day operating period, AOB in a continuous pre-denitrification reactor did not adapt to the FNA that was applied to treat some of the return activated sludge. In contrast, NOB did adapt to FNA. NOB activity in the seed sludge was only 11% of the original activity after FNA batch treatment, at 0.75mg HNO2-N/L. NOB activity in the pre-denitrification reactor was not affected after being exposed to this FNA level. Nitrosomonas was the dominant AOB before and after long-term FNA treatment. However, dominant NOB changed from Nitrospira to Candidatus Nitrotoga, a novel NOB genus, after long-term FNA treatment. This adaptation of NOB to FNA may be due to the shift in NOB population makeup.
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