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•A unique feature of the SNAD in this study was using suspended activated sludge.•Only 1.02 mg/L of nitrate was detected in the effluent treating real domestic sewage.•TN removal ...efficiency of 86.1% was achieved in the SNAD system.•Micro-aeration saved energy as well as maintained the efficiency of the system.•Quantity of Candidatus Brocadia and Thauera increased in the SNAD system.
Bottleneck issues such as difficult biofilm culturing, time-consuming granular formation and excessive nitrate production, always limited the application of anammox processes. By using the common but meaningful suspended activated sludge instead of biofilm or granule activated sludge, a combined anammox process, simultaneous partial nitrification, anammox and denitrification (SNAD), was established in a sequencing batch reactor (SBR) in this study to treat real domestic sewage and was operated for 300 days. Results indicated that a high ammonia removal rate and nitrogen removal rate (1.1 and 1.03 (kgN/(m3·d)), respectively) were achieved during the baseline phase treating high ammonia wastewater (350–550 mg/L). During the experimental phase treating domestic sewage with a C/N of 3–3.5, 86.1% of total nitrogen removal efficiency was achieved and only 1.02 mg/L of nitrate concentration was detected in the effluent with an averagely 64.6 mg/L of ammonium in the influent. Nitrogen mass balance analysis demonstrated that about 89% of total nitrogen (TN) was removed by anammox and the remaining 11% of TN was removed by denitrification. Bacterial abundance of Candidatus Brocadia and Thauera increased from 0.02% to 0.5% and 0.39% to 2.24% (from baseline phase to experimental phase), respectively, which made an essential contribution to the nitrogen removal. The advanced nitrogen removal performance reached in the SNAD process showed the possibility of using suspended sludge as a substitute of biofilm and granule sludge concerning anammox, and the process could treat real domestic sewage with a C/N of 3–3.5 without pretreatment of chemical oxygen demand (COD).
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•Continuous aeration in membrane-aerated biofilms resulted in full nitrification.•Intermittent aeration suppressed nitrite oxidation and supported anammox process.•Biofilm pH ...presented periodic upshifts with aeration switches.•Free ammonia speciation likely caused the inhibition of nitrite-oxidizing bacteria.•Heterotrophs established nitrous oxide-reduction zones under intermittent aeration.
Membrane-aerated biofilm reactors (MABR) are being applied for autotrophic nitrogen removal, yet control of nitrogen turnover remains challenging in MABR counter-diffusion biofilms. In this study, we regulated microbial activities in two lab-scale MABRs by providing continuous versus intermittent aeration. Nitrogen consumption by different functional microbial groups was estimated from bulk measurements via a mass balance approach. Nitrite-oxidizing bacteria (NOB) proliferated under continuous aeration while they were significantly suppressed under intermittent aeration, and NOB suppression activated anaerobic ammonium oxidation. Nitritation performance in the MABR was studied through long-term bulk measurements and in situ biofilm microprofiles of dissolved oxygen (DO) and pH. During intermittent aeration pH effects rather than DO effects determined nitritation success, especially ammonia speciation, which serves as substrate and inhibitor in nitrification processes. Biofilm transition phases were monitored upon aeration switches. Canonical correspondence analysis suggested that the relative transition after anoxia and aeration intermittency were less decisive for biofilm performance than the relative aeration duration. Heterotrophic bacteria displayed minor denitrification rates with aeration control, but contributed to mitigation of nitrous oxide (N2O) emissions. N2O production hotspots were identified at the top of the anoxic biofilm zone under continuous aeration. Instead, under intermittent aeration an anoxic N2O reduction zone was established. Our observations support intermittent aeration control of MABRs as a simple strategy for energy-efficient nitrogen removal with low N2O emission. .
Nobiletin (Nob) is a major component among the most reported polymethoxyflavones (PMFs), which possesses multiple efficacious healthcare activities. Owing to its high melting point and poor water ...solubility, the oral bioavailability of Nob needs to be improved via loading Nob on carriers. To take full advantage of Nob, the interaction mechanism between Nob and vehicles should be clarified. Herein, β-lactoglobulin (β-LG) was selected as the vehicle and further investigated the binding mechanism between Nob and β-LG. The binding stoichiometry of complex was found to be 1:1 by analysis of intrinsic fluorescence experiment. The results also confirmed by isothermal titration calorimetry (ITC) measurement that the binding behavior between β-LG and Nob was a spontaneously endothermic process driving mainly by hydrophobic interaction. Moreover, competitive binding and molecular docking method indicated the Nob was primary bound to internal calyx of β-LG at neutral pH. UV spectrophotometry revealed that the solubility of Nob was enhanced to 3 times by forming complex. Furthermore, Nob could alter secondary structure of β-LG by a transition from α-helix to β-sheet and lead to small increase on surface hydrophobicity of β-LG. This work will provide some valuable information on clarifying the interaction between protein and PMFs, which contributing to improve the poor bioavailability of PMFs.
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•Nobiletin had influence on the secondary and tertiary structure of β-lactoglobulin.•The hydrophobic interaction played a vital role in the interaction between Nobiletin and β-lactoglobulin.•The formation of β-lactoglobulin/Nobiletin complex was a spontaneously endothermic process.•At neutral pH, Nobiletin was primary bound into internal calyx of β-lactoglobulin.•Nobiletin solubility could be improved up to 3 times by forming β-lactoglobulin/Nobiletin complex.
Sidestream sludge treatment approaches have been developed in recent years to achieve mainstream nitrite shunt or partial nitritation, where NOB are selectively inactivated by biocidal factors such ...as free nitrous acid (FNA) or free ammonium (FA) in a sidestream reactor. The existence of NOB in raw wastewater has been increasingly realized and could pose critical challenge to stable NOB suppressions in those systems. This study, for the first time, evaluated the impact of influent NOB on the NOB suppressions in a mainstream nitrite shunt system achieved through sidestream sludge treatment. An over 500-day sequential batch reactor operation with six experimental phases rigorously demonstrated the negative effects of influent NOB on mainstream NOB control. Continuously seeding of NOB contained in influent stimulated NOB community shifts, leading to different extents of ineffective NOB suppression. The role of primary wastewater treatment in NOB removal from raw wastewater was also investigated. Results suggest primary settling and High Rate Activated Sludge system could remove a large part of NOB contained in raw wastewater. Primary treatment for raw wastewater is necessary for ensuring stable mainstream NOB suppressions.
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•Influent NOB challenge NOB suppression by stimulating community shift and resistance.•Primary settling substantially reduces NOB in raw wastewater.•High-rate activated sludge treatment substantially reduces NOB in raw wastewater.•This study expanded the current knowledge of mainstream NOB suppression.
The partial nitritation-anammox (PNA) process is the most promising technique to treat municipal sewage; however, nitrite oxidizing bacteria (NOB) are a hindrance to achieve PNA. This study ...investigated the effects of selectively discharging flocs (<200 μm) to washout NOB in a sequencing batch reactor (SBR) over 200 d. The experiment was divided into three phases with different floc sludge retention times (SRTs; 30, 20 and 30 d). When the SRT of the flocs was reduced from 30 to 20 d to washout NOB, a significant reduction of ammonia oxidizing bacteria (AOB) and anaerobic ammonium-oxidizing (anammox) bacteria in the flocs was found. This indicates that a low floc SRT (20 d) leads to the loss of AOB and anammox bacteria in the flocs (<200 μm) and destroys PNA. Activity tests and qPCR analysis revealed the variations of functional bacteria in the granules and flocs, indicating that the enrichment of AOB, NOB, anammox bacteria in the granules is caused by the long-term discharging of flocs. High-throughput sequencing analysis revealed that the microbial shift of Tetrasphaera was significant in the flocs and may be connected to the enrichment of anammox bacteria and the stability of the PNA requires further research. All the obtained NOB sequences were affiliated with the genera Nitrospira and could further influence the PNA system. Overall, this study provides an in-depth understanding of the impact of discharging flocs to washout NOB and promotes the application of combing granules/floc PNA in sewage treatment.
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•Controlling floc SRTs of 30 d was beneficial to stable operation of PNA.•Floc SRTs of 20 d would result in the reduction of AOB and Anammox in flocs.•The distribution of NOB in granules and flocs was impacted by long-term discharging flocs.•The microbial community structure was seriously affected by discharging flocs.
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•A novel three-stage process, comprising PN/A, acidic PN, and anammox, is proposed.•The PN/A unit requires residual ammonium to suppress nitrite-oxidizing bacteria (NOB).•The acidic ...PN unit provides stable nitrite sources to the anammox unit.•The anammox unit removes the residual ammonium and nitrite from the PN/A and the acidic PN units.•The overall nitrogen removal efficiency is higher than 90% based on model simulation.
Sewage treatment with partial nitritation and anammox (PN/A, also termed as deammonification) has gained widespread attentions over the past decade, driven by the need of establishing energy-neutral/positive sewage treatment plants (STPs). However, a key challenge for the mainstream deammonification is to achieve stable suppression of nitrite-oxidizing bacteria (NOB). Deammonification can be configured in either one reactor or two reactors (where PN and anammox are separated), known as one-stage and two-stage configurations, respectively. This study first analyses the current bottlenecks in one-stage and two-stage deammonification. To address these issues, a new, three-stage configuration is proposed. Three-stage deammonification comprises three components, including a PN/A reactor, an acidic PN reactor, and an anammox reactor. The majority of sewage is allocated to the PN/A reactor, which produces an effluent with an elevated level of residual ammonium (2–40 mg N L−1), as required for NOB suppression in this reactor. The residual ammonium is removed in the anammox reactor, using the nitrite supplied from the acidic PN reactor. The acidic PN reactor is fed with a mixture of sewage and anaerobic digestion liquor with a ratio that allows the attainment of a low pH (<6.0) and NOB suppression. This three-stage process was shown applicable to a wide range of municipal wastewaters which have a HCO3–/ NH4+ mole ratio of 1.17–2.41, based on mass balance. Further, simulation indicates that this three-stage process can deliver a stable and high-quality effluent, with the total nitrogen < 10 mg N L−1 and the nitrogen removal efficiency > 90%.
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•Aerobic granules SNDPR used for hypersaline wastewater treatment.•Reliable nitrogen removal efficiency was achieved at elevated salinity.•EBPR was deteriorated with 2 % (w/v) ...salinity stress.•Salinity induced changes in granular community structure.•Halophilic and non-halophilic bacteria were recognized.
Hypersaline wastewater may pose threats to biological wastewater treatment processes. An aerobic granular sludge-based sequencing batch reactor (SBR) performing simultaneous nitrification, denitrification and phosphorus removal (SNDPR) was evaluated with increased salinity from 1 to 2 % (w/v). Nitrogen removal performance was unaffected by salinity up to 20 g/L in terms of reliable and efficient nitrification and denitrification. Enhanced biological phosphorus removal (EBPR) process was completely deteriorated at salinity up to 2 %, in contrast to excellent phosphorus removal at 1 %. Profiles of phosphorus over one cycle demonstrated that higher salinity not only inhibited anaerobic phosphorus release but also impeded aerobic/anoxic phosphorus uptake. Illumina MiSeq sequencing revealed multiple halophilic and non-halophilic bacteria within aerobic granules with family Anaerolineaceae being the predominant potential salt adapter. Besides, ammonia oxidizing bacteria (AOB), glycogen accumulating organisms (GAOs) were more tolerant to salt than nitrite oxidizing bacteria (NOB) and phosphorus accumulating organisms (PAOs) and denitrifying PAOs (DNPAOs). These results deciphered the resilience of aerobic granular sludge-based biological nitrogen and phosphorus removal processes to hypersaline stress.
The accumulation of nitrite is frequently overlooked, despite the fact that nitrification is the most essential phase of the entire nitrogen (N) cycle and that nitrifying bacteria play a significant ...role in nitrification. At present, the effects of different N application rates on soil nitrite-oxidizing bacteria (NOB) abundance, community composition, diversity, and its main influencing factors are still unclear. In this study, five N fertilizer application rates under film mulching and a drip irrigation system were studied in the semi-arid area of Northeast China. The treatments were 0 kg ha−1 (N0), 90 kg ha−1 (N1), 150 kg ha−1 (N2), 210 kg ha−1 (N3), and 270 kg ha−1 (N4). Fluorescent quantitative PCR and Illumina Miseq sequencing were used to analyze the abundance and community structure of NOB under different amounts of N application. The results showed that the increase in amounts of N application was strongly accompanied by an increase in the content of soil organic matter (SOM), total nitrogen (TN), nitrate nitrogen (NO3−-N), and ammonium nitrogen (NH4+-N), while the pH significantly reduced with an increase in N fertilization. N fertilization significantly increased soil nitrite oxidoreductase (NXR) activity, soil nitrification potential (PNR), and soil nitrite oxidation potential (PNO). A high N application rate significantly heightened the abundance of Nitrospira- and Nitrobacter-like NOB. N fertilizer considerably raised the Shannon index of Nitrospira-like NOB. The N application amount was the key factor affecting the community structure of Nitrospira-like NOB, and available nitrogen (AN) had the dominant influence on the community structure of Nitrospira-like NOB. N fertilizer can cause soil acidification, which affects NOB abundance and diversity. Nitrospira-like NOB may promote nitrite oxidation in different N application rates under a mulched fertigation system. The findings offered a crucial scientific foundation for further investigation into how nitrite-oxidizing bacteria respond to N fertilizer management strategies in farmland soil under film mulching drip irrigation in Northeast China.
Nitrifiers are the key player in the nitrogen cycle of agroecosystems, yet less research has focused on their performance and response in saline ecosystems. In this study, we carried out potting ...experiments with biochar and wood vinegar as saline soil amendments under rice cultivation conditions with four different treatments: without biochar or wood vinegar (CK), biochar (BC), wood vinegar (WV), and biochar + wood vinegar (BC + WV). The results showed that the addition of biochar and/or wood vinegar decreased the soil pH and electrical conductivity (EC), which led to an increase in the gene abundance of ammonia-oxidizing bacteria (AOB), thereby benefiting the advancement of the potential nitrification rate (PNR). WV and BC + WV significantly increased the gene abundance of Nitrospira. In addition, the addition of biochar and wood vinegar altered the community composition of ammonia-oxidizing archaea (AOA) and ammonia-oxidizing bacteria (AOB), while the NH4+-N content was the key factor affecting the nitrifier communities. Compared to the CK group, biochar and/or wood vinegar significantly increased the relative abundance of Nitrosospira cluster 3 b in AOB and unknown affiliation in nitrite-oxidizing bacteria (NOB). Overall, the abundance and community composition of AOB contributed more to the PNR than those of AOA, while NOB played a pivotal role in the potential nitrite oxidation (PNO) rate in sodic saline soils. In conclusion, the addition of biochar with wood vinegar had positive effect on improving sodic saline soils by improving the physicochemical properties of the soils, increasing the abundance of nitrifier and changing the community structure of nitrifier. Exploration of the key drivers of soil nitrifier processes is potentially useful for understanding the biological potential of nutrient cycling, providing novel insight into the effects of human intervention and soil management.
The mainstream partial nitritation/anammox (PN/A) process has been intensively studied but its stability remains a key challenge. It is shown here that biofilm thickness can exhibit a critical role ...in controlling the process stability of mainstream PN/A against dissolved oxygen (DO) variation. In a laboratory moving bed biofilm reactor (MBBR), PN/A performance was initially established in 200 days by controlling a low DO of 0.13 ± 0.07 mg O2/L in the bulk liquid, which deteriorated with an increase of DO (0.35 ± 0.13 mg O2/L) for over two months, and then rapidly recovered in a month with the initial low DO level re-applied. Biofilm thickness of PN/A carriers was measured during the experiment, which became significantly thinner (367 ± 146 μm) at mainstream conditions. The thin thickness primarily decreased the in-situ consumption rate of nitrite, rather than ammonium, when DO increased from 0.1 to 0.4 mg O2/L, due to that the thin thickness can only restrict anammox capacity. These results illustrated the role of biofilm thickness in regulating PN/A performance and microbial activities. Further investigation using an established model revealed the joint contribution of biofilm thickness and DO concentration to PN/A process, while particularly, the biofilm thickness can determine the optimal DO level for maximizing the nitrogen removal efficiency and system robustness against DO variation. These results highlight the need of considering biofilm thickness in PN/A process optimization and stability improvement in low-strength wastewater treatment.
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•N-removal efficiency >80% is achieved via PN/A in a mainstream MBBR.•The mainstream PN/A is stably realized at DO concentration of 0.13 ± 0.07 mgO2/L.•Biofilm thickness controls the maximal capacity of anammox rather than AOB or NOB.•A narrow DO range (0.15-0.20 mgO2/L) is required for high N-removal in thin biofilm.
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