Ultra-high nitrogen removal from real municipal wastewater using selective enhancement of glycogen accumulating organisms (GAOs) in a partial nitrification-anammox (PNA) system.

Integrating endogenous denitrification (ED) into partial nitrification-anammox (PNA) systems by adequately utilizing organics in municipal wastewater is a promising approach to improve nitrogen removal efficiency (NRE). In this study, a novel strategy to inhibit phosphorus-accumulating organisms (PAOs) by inducing phosphorus release and exclusion was adopted intermittently, optimizing organics allocation between PAOs and glycogen-accumulating organisms (GAOs). Enhanced ED-synergized anammox was established to treat real municipal wastewater, achieving an NRE of 97.5±2.2% and effluent total inorganic nitrogen (TIN) of less than 2.0 mg/L. With low poly-phosphorus (poly-P) levels (poly-P/VSS below 0.01 (w/w)), glycogen accumulating metabolism (GAM) acquired organics exceeded that of phosphorus accumulating metabolism (PAM) and dominated endogenous metabolism. Ca. Competibacter (GAO) dominated the community following phosphorus-rich supernatant exclusion, with abundance increasing from 3.4% to 5.7%, accompanied by enhanced ED capacity (0.2 to 1.4 mg N/g VSS /h). The enriched subgroups (GB4, GB5) of Ca. Competibcater established a consistent nitrate cycle with anammox bacteria (AnAOB) through endogenous partial denitrification (EPD) at a ∆NO2--N/∆NH4+-N of 0.91±0.11, guaranteeing the maintenance of AnAOB abundance and performance. These results provide new insights into the flexibility of PNA for the energy-efficient treatment of low-strength ammonium wastewater.

Data fusion of electronic noses and electronic tongues aids in botanical origin identification on imbalanced Codonopsis Radix samples.

Codonopsis Radix (CR) is an edible food and traditional Chinese herb medicine in China. Various varieties of Codonopsis Radix have different tastes. To make the flavor of processed food stable, two kinds of electronic sensory devices, electronic nose and electronic tongue, were used to establish a discrimination model to identify the botanical origin of each sample. The optimal model built on the 88 batches of samples was selected from the models trained with all combination of two pretreatment methods and three classification methods. A comparison were performed on the models trained on the data collected by electronic nose and electronic tongue. The results showed that the model trained on the fused dataset outperformed the models trained separately on the electronic nose data and electronic tongue data. The two preprocessing approaches could improve the prediction performance of all classification methods. Classification and Regression Tree approach performed better than Partial Least Square Discriminant Analysis and Linear Discriminant Analysis in terms of accuracy. But Classification and Regression Tree tends to assign the samples of minority class to the majority class. Meanwhile, Partial Least Square Discriminant Analysis keeps a good balance between the identification requirements of all the two groups of samples. Taking all the results above, the model built using the Partial Least Square Discriminant Analysis method on the fused data after z-score was used to identify the botanical origin of Codonopsis Radix.

Simultaneous nitrogen and phosphorus removal from municipal wastewater by Fe(III)/Fe(II) cycling mediated partial-denitrification/anammox.

The efficient removal of nitrogen and phosphorus remains challenging for traditional wastewater treatment. In this study, the feasibility for enhancing the partial-denitrification and anammox process by Fe (III) reduction coupled to anammox and nitrate-dependent Fe (II) oxidation was explored using municipal wastewater. The nitrogen removal efficiency increased from 75.5 % to 83.0 % by adding Fe (III). Batch tests showed that NH4+-N was first oxidized to N2 or NO2--N by Fe (III), then NO3--N was reduced to NO2--N and N2 by Fe (II), and finally, NO2--N was utilized by anammox. Furthermore, the performance of phosphorus removal improved by Fe addition and the removal efficiency increased to 78.7 %. High-throughput sequencing showed that the Fe-reducing bacteria Pseudomonas and Thiobacillus were successfully enriched. The abundance of anammox bacterial increased from 0.03 % to 0.22 % by multiple nitrite supply pathways. Fe addition presents a promising pathway for application in the anammox process.

[Critical quality attribute characterization of big brand TCM: taste formulation optimization strategy and application of Xiaoer Ganmao Oral Liquid based on taste quality attribute].

The taste is the quality attribute for the development and production of traditional Chinese medicine(TCM). To improve the medication compliance of the big brand TCM, Xiaoer Ganmao Oral Liquid, a correlation model between the electronic tongue sensor signal value and human sensory evaluation score was established, and an optimization strategy of taste improvement for Xiaoer Ganmao Oral Liquid was developed with the key techniques of statistical experimental design. Based on the above model, the optimal formulation was determined as follows: aspartame content of 1-2 mg·mL~(-1), acesulfame-K content of 1.5-3 mg·mL~(-1), and steviol glycoside content of 1-2 mg·mL~(-1). Furthermore, the optimal formulation was verified by human sensory evaluation. Therefore, the taste of Xiaoer Ganmao Oral Liquid was improved. Taking Xiaoer Ganmao Oral Liquid as an example, the present study developed the taste formulation optimization method based on the correlation between the electronic tongue and human sensory evaluation, which is expected to provide an important reference to improve the taste of oral liquid of TCM.

Enhanced nutrient removal from mainstream sewage via denitrifying dephosphatation, endogenous denitrification and anammox in a novel continuous flow process.

It is a challenging subject to realize nitrogen and phosphorus elimination synchronously from limited-carbon sewage through conventional biological processes. Herein, a novel continuous flow anaerobic/oxic/anoxic/oxic (AOA-O) process, which integrated denitrifying dephosphatation, endogenous denitrification and anammox in the anoxic zone, was developed to enhance nutrient elimination from low carbon/nitrogen sewage (3.4 in average). After the long-term operation (280 days), a satisfactory nutrient removal performance (effluent PO43--P: 0.2 mg P/L, total inorganic nitrogen (TIN):8.9 mg N/L) was obtained. Mass balance indicated that anammox contributed to 26.1% TIN removal and denitrifying dephosphatation contributed to 25.6% phosphorus removal, respectively. The cooperation of anammox bacteria retained in biofilms and endogenous denitrifying bacteria in flocculent sludge was responsible for the enhanced nutrient removal in the anoxic zone. Dechloromonas carried out phosphorus uptake both under oxic conditions and anoxic conditions. This study can broaden the application prospect of mainstream anammox.

Anaerobic duration optimization improves endogenous denitrification efficiency by glycogen accumulating organisms enhancement.

Without additional carbon sources, a low endogenous denitrification rate (EDNR) is the critical factor limiting its application in postdenitrification systems. This study optimized the quantitative distribution of anaerobic carbon source removal pathways based on chemometrics for the first time and explored the effect of anaerobic carbon conversion on anoxic endogenous denitrification. Results showed that enhancing the intracellular carbon storage of glycogen accumulating organisms (GAOs) by optimizing anaerobic duration can effectively improve the EDNR. The anaerobic stage was proposed to end at the peak concentration of polyhydroxyalkanoates (PHAs). A two-stage endogenous denitrification system was established to explore the long-term operating performance before and after optimizing anaerobic duration. Results showed that the average NO3- removal rate increased by 25%. qPCR and optimized stoichiometric analyses indicated that the relative abundance and intracellular carbon storage proportion of GAOs increased by 67% and 25%, respectively. This study provided an effective strategy to improve postdenitrification efficiency.

[Critical quality attribute assessment of big brand traditional Chinese medicine: study on NIR field detection method of chemical properties of moisture in Tongren Niuhuang Qingxin Pills].

For the field detection problems of critical quality attribute(CQA) of moisture content in traditional Chinese medicine(TCM) manufacturing process, big brand TCM Tongren Niuhuang Qingxin Pills were used as the carrier, to establish a moisture content NIR field detection model with or without cellophane in real world production with use of near infrared(NIR) spectroscopy combined with stoichiometry. With the moisture content determined by drying method as reference value, the partial least square method(PLS) was used to analyze the correlation between the spectrum and the moisture reference value. Then the spectral pretreatment methods were screened and optimized to further improve the accuracy and stability of the model. The results showed that the best quantitative model was developed by the spectral data pretreatment of standard normal variate(SNV) with the latent variable factor number of 2 and 7 of Tongren Niuhuang Qingxin Pills with or without cellophane samples. The prediction coefficient of determination(R_(pre)~2) and standard deviation of prediction(RMSEP) of the model with cellophane samples were 0.765 7 and 0.157 2%; R_(pre)~2 and RMSEP of the model without cellophane samples were 0.772 2 and 0.207 8%. The NIR quantitative models of moisture content of Tongren Niuhuang Qingxin Pills with and without cellophane both showed good predictive performance to realize the rapid, accurate and non-destructive quantitative analysis of moisture content in such pills, and provide a method for the field quality control of the critical chemical attributes of moisture in the manufacturing of big brand TCM.

[Critical quality attribute assessment of big brand traditional Chinese medicine: quality control of Tongren Niuhuang Qingxin Pills in texture and sensory attributes].

Texture sensory attributes are the key items in quality control of Chinese medicinal honeyed pills. The purpose of this study is to develop a quality control method for assessing the texture sensory attributes of Chinese medicinal honeyed pills based on real-world Tongren Niuhuang Qingxin pilular masses and finished products. First, parameters of texture profile analysis(TPA) were optimized through single factor and central composite design(CCD) experiments to establish a detection method for texture sensory attri-butes of Tongren Niuhuang Qingxin Pills. The results showed that the established detection method was stable and reliable, with the optimal parameters set up as follows: deformation percentage of 70%, detection speed at 30 mm·min~(-1), and interval time of 15 s. Furthermore, 540 data points yielded form six texture sensory attributes of pills from 30 batches were subjected to multivariate statistical process control(MSPC) with Hotelling T~2 and squared prediction error(SPE) control charts to establish the quality control method of Tongren Niuhuang Qingxin Pills. This study is expected to provide a reference for improving the quality control system of Chinese medicinal honeyed pills.

A draft genome for Spatholobus suberectus.

Spatholobus suberectus Dunn (S. suberectus), which belongs to the Leguminosae, is an important medicinal plant in China. Owing to its long growth cycle and increased use in human medicine, wild resources of S. suberectus have decreased rapidly and may be on the verge of extinction. De novo assembly of the whole S. suberectus genome provides us a critical potential resource towards biosynthesis of the main bioactive components and seed development regulation mechanism of this plant. Utilizing several sequencing technologies such as Illumina HiSeq X Ten, single-molecule real-time sequencing, 10x Genomics, as well as new assembly techniques such as FALCON and chromatin interaction mapping (Hi-C), we assembled a chromosome-scale genome about 798 Mb in size. In total, 748 Mb (93.73%) of the contig sequences were anchored onto nine chromosomes with the longest scaffold being 103.57 Mb. Further annotation analyses predicted 31,634 protein-coding genes, of which 93.9% have been functionally annotated. All data generated in this study is available in public databases.

Combining simultaneous nitrification-endogenous denitrification and phosphorus removal with post-denitrification for low carbon/nitrogen wastewater treatment.

Due to the limited nutrient removal from low carbon/nitrogen (⩽4) wastewater, a process combined simultaneous nitrification-endogenous denitrification and phosphorus removal (SNDPR) with post-denitrification (PD) in a SBR was proposed for deep-level nutrient removal without external carbon addition. SNDPR driven by PAOs and GAOs reduced PO4(3-)-P (98.3%) and partial TN (59.0%) at low DO conditions (0.5±0.1mg/L), and post-dentrification achieved further NOX(-) (produced by SNDPR) removal (24.0%) anoxically by utilizing the residual intracellular polymers in GAOs. Combined control of anaerobic/aerobic/anoxic durations and low DO inhibition to aerobic GAOs and NOB conducted partial nitrification-endogenous denitrification (PNED) (66%), which saved 44.3% intracellular polymers to further reduce 64% TN in effluent. After 115-day operation, the average effluent PO4(3-)-P and TN concentrations were 0.4 and 3.9mg/L, respectively, with 92.1% of TN removal. Highly enriched PAOs (36%±2%), GAOs (22%±2%) and AOB (15%±3%) over NOB (3%±1%) facilitated P uptake, PNED and post-denitrification in the SNDPR-PD system.

Enhancement of denitrifying phosphorus removal and microbial community of long-term operation in an anaerobic anoxic oxic-biological contact oxidation system.

A two-sludge system consisting of anaerobic anoxic oxic-biological contact oxidation (A(2)/O-BCO) was developed to treat domestic wastewater with a low carbon/nitrogen (COD/TN) ratio (around 3.21) by shortening sludge retention time (SRT) for phosphorus accumulating organisms (PAOs) in the A(2)/O reactor and prolonging SRT for nitrifiers in the BCO reactor. Specifically, the BCO reactor was composed of three stages in series (N1, N2 and N3), so that simultaneous nitrogen and phosphorus removals by denitrifying PAOs (DNPAOs) was achieved in the A(2)/O reactor with [Formula: see text] as the electron acceptor from the BCO reactor. Long term operational tests (600 days) were conducted with various operational parameters [e.g., hydraulic retention time (HRTs), nitrate recycling ratio (Rs), volume ratio (Vs)] to examine the denitrifying phosphorus removal performance. The system exhibited the highest removal of TN and [Formula: see text] at the HRTs of 8 h, Rs of 300% and Vs of 2:4:1. The optimal TN and [Formula: see text] removals were 80.30% and 96.61% at low COD/TN of 3.21. The species diversity and microbial community examined by the Illumina MiSeq method demonstrated the fact of two-sludge system, and the improved community structure by long-term optimization was prominent comparing with the seed sludge. Additionally, Accumulibacter and Dechloromonas were the dominant functional PAOs with 25.74% in the A(2)/O reactor, while nitrifiers (including Nitrosomonas and Nitrospira) were gradually enriched with 13.10%, 21.33%, and 31.10% in the three stages of the BCO reactor.

A novel stoichiometries methodology to quantify functional microorganisms in simultaneous (partial) nitrification-endogenous denitrification and phosphorus removal (SNEDPR).

Although efficient removal of carbon (C), nitrogen (N) and phosphorous (P) from wastewater with low C/N ratio was achieved in anaerobic/aerobic simultaneous nitrification-endogenous denitrification and phosphorus removal (SNEDPR) systems, the removal pathways and metabolic transformations in this complex system are unclear. This work targeted at developing the stoichiometric models for denitrifying glycogen organisms (DGAOs) via nitrite and nitrate (DGAONi and DGAONa), and demonstrating a novel methodology to quantify diverse functional microorganisms (e.g. ammonia and nitrite oxidizing bacteria, aerobic phosphorus accumulating organisms (APAOs), denitrifying PAOs (DPAOs) and aerobic GAOs (AGAOs)) for the removal of C, N and P. The results showed that the anaerobic intracellular carbon storage (CODintra) was mainly accomplished by GAOs, and PAOs were only responsible for about 40% of CODintra through a stable P release. At the aerobic stage, 84.9% of P was removed by APAOs with 15.1% left by DPAOs, while 64.6% of N was removed by DGAOs (45.8% by DGAONi and 18.8% by DGAONa) with 18.1% by DPAOs and 17.3% by bacterial growth. High proportion of N removal via nitrite (partial nitrification-endogenous denitrification) (71%) saved 7.3% aeration and 38% intracellular carbon demand. However, AGAOs still activated well at the aerobic intercellular carbon consumption, which limited the further improvement of N removal efficiency. By elucidating the nutrient removal pathways among diverse functional microorganisms, the methodology developed in this study could accelerate the nutrient removal in the SNEDPR process.

Enhanced volatile fatty acids production of waste activated sludge under salinity conditions: Performance and mechanisms.

Volatile fatty acids (VFAs) are essential for removing biological nitrogen and phosphorus in wastewater treatment plants. The purpose of this work was to investigate whether and how the addition of NaCl could improve the production of VFAs from waste activated sludge (WAS). Sludge solubilization was efficiently improved by the addition of NaCl. Both protein and carbohydrate in the fermentation liquid increased with the dosage of NaCl, and it provided a larger amount of organic compounds for the production of the VFAs. NaCl had inhibitory effects on the production of methane and a high dosage of NaCl could severely suppress the growth of methanogens, which decreased the consumption of the VFAs. Consequently, the production of VFAs was significantly enhanced by the addition of NaCl. The maximum production of VFAs was achieved with the highest dosage of NaCl (3316 mg (COD)/L at the NaCl dosage 0.5 mol/L; 783 mg (COD)/L without the addition of NaCl). Therefore, this study indicates that using NaCl could be an efficient method for improving the production of VFAs from WAS.

Treating low carbon/nitrogen (C/N) wastewater in simultaneous nitrification-endogenous denitrification and phosphorous removal (SNDPR) systems by strengthening anaerobic intracellular carbon storage.

A novel simultaneous nitrification denitrification and phosphorous removal-sequencing batch reactor (SNDPR-SBR) enriched with PAOs (phosphorus accumulating organisms), DPAOs (denitrifying PAOs), and GAOs (glycogen accumulating organisms) at the ratio of 2:1:1 was developed to achieve the simultaneous nutrient and carbon removal treating domestic wastewater with low carbon/nitrogen ratio (≤3.5). The SNDPR system was operated for 120 days at extended anaerobic stage (3 h) and short aerobic stage at low oxygen concentration (2.5 h) with short sludge retention time (SRT) of 10.9 d and hydraulic retention time (HRT) of 14.6 h. The results showed that at the stable operating stage, the average effluent chemical oxygen demand (COD) and PO4(3-)-P concentrations were 47.2 and 0.2 mg L(-1), respectively, the total nitrogen (TN) removal efficiency was 77.7%, and the SND efficiency reached 49.3%. Extended anaerobic stage strengthened the intracellular carbon (mainly poly-ß-hydroxybutyrate, PHB) storage, efficiently utilized the organic substances in wastewater, and provided sufficient carbon sources for denitrification and phosphorus uptake without external carbon addition. Short aerobic stage at low oxygen concentration (dissolved oxygen (DO): 1 ± 0.3 mg L(-1)) achieved a concurrence of nitrification, endogenous denitrification, denitrifying and aerobic phosphorus uptake, and saved about 65% energy consumption for aeration. Microbial community analysis demonstrated that P removal was mainly performed by aerobic PAOs while N removal was mainly carried out by denitrifying GAOs (DGAOs), even though DPAOs were also participated in both N and P removal.

[Role of PI3K/Akt pathway in effect of paeoniflorin against Aß25-35-induced PC12 cell injury].

OBJECTIVE: To study the role of PI3K/Akt pathway in the neuroprotective effect of paeoniflorin on PC12 cells. METHOD: The paeoniflorin group (5, 10, 20 µmol · L(-1)) was pretreated for 30 min, and then added with Aß25-35 (20 µmol · L(-1)) for interaction for 24 h. Inhibitor LY294002 (10 µmol · L(-1)) was pretreated for 30 min before the action of paeoniflorin (10 µmol · L(-1)). The MTT colorimetric method was used to detect the cell viability. The apoptosis rate was tested by the FITC-Annexin V/PI staining. The protein expression of p-AKT, Bax, Bcl-2 and cleaved caspase-3 protein were detected by Western blot analysis. RESULT: Paeoniflorin could significantly inhibit the Aß25-35-induced PC12 cell toxicity and apoptosis. Its protection effect may be achieved by up- regulating AKT phosphorylation level, increasing Bcl-2 protein expression, reducing Bax protein expression, inhibiting the activation of caspase-3. Inhibitor LY294002 could weaken the above protective effects of paeoniflorin. CONCLUSION: Paeoniflorin could activate PI3K/Akt signaling pathway to protect the PC12 cell injury induced by Aß25-35.

[Protective effects of paeoniflorin against aBETA25-35-induced oxidative stress in PC12 cells].

OBJECTIVE: To investigate the neuroprotective effects and mechanism of paeoniflorin on Abeta25-35-induced oxidative stress in PC12 cells. METHOD: The proliferation of induced PC12 cells were investigated by the MTT method. The leakage rate of lactate dehydrogenase (LDH), the intracellular content of malondialdehyde (MDA), superoxide dismutase (SOD) and glutathione peroxidase (GSH-Px) were also measured. The changes of the intracellular reactive oxygen species (ROS) level and the mitochondrial membrane potential (MMP) were investigated by flow cytometry using DCFH-DA and Rh123 staining. The protein expression of HO-1, Cyt C and cleaved Caspase-3 was detected by Western blot. RESULT: The results showed that paeoniflorin at different doses (5, 10, 20 micromol . L-1) could increase the cell viability and activities of antioxidant enzyme (GSH-Px, SOD and HO-1), prevent LDH release and lipid peroxidation (MDA) production, decrease the level of intracellular ROS, increase MMP, inhibit the release of cytochrome c from the mitochondria and attenuate activation of Caspase-3. CONCLUSION: Paeoniflorin can protect PC12 cells from Abeta25-35 oxidative stress injuries. The mechanism may be related to its antioxidant action and inhibition of mitochondria-mediated caspase signaling pathways.

Achieving nitritation and phosphorus removal in a continuous-flow anaerobic/oxic reactor through bio-augmentation.

The feasibility of achieving nitritation and phosphorus removal using bio-augmentation was investigated in a continuous-flow anaerobic/oxic (A/O) reactor treating sewage. The results indicated that nitritation could be quickly start-up, and reconstructed with an increase in the nitrite accumulation rate (NAR) from 1% to 89% within 15 days by using bio-augmentation and controlling DO at 0.96 mg/L. Biological phosphorus removal could be achieved with the average phosphorus removal efficiency of 96.43% when the NAR was maintained above 78.60%. Meanwhile, sludge settleablity was good with a sludge volume index (SVI) of between 62 and 102 mL/g even under high NAR. After nitritation and biological phosphorus removal were achieved, this A/O reactor has the potential to supply appropriate influent for the anammox UASB reactor.

The truncated C-terminal RNA recognition motif of TDP-43 protein plays a key role in forming proteinaceous aggregates.

TDP-43 is the major pathological protein identified in the cellular inclusions in amyotrophic lateral sclerosis and frontotemporal lobar degeneration. The pathogenic forms of TDP-43 are processed C-terminal fragments containing a truncated RNA-recognition motif (RRM2) and a glycine-rich region. Although extensive studies have focused on this protein, it remains unclear how the dimeric full-length TDP-43 is folded and assembled and how the processed C-terminal fragments are misfolded and aggregated. Here, using size-exclusion chromatography, pulldown assays, and small angle x-ray scattering, we show that the C-terminal-deleted TDP-43 without the glycine-rich tail is sufficient to form a head-to-head homodimer primarily via its N-terminal domain. The truncated RRM2, as well as two ß-strands within the RRM2, form fibrils in vitro with a similar amyloid-negative staining property to those of TDP-43 pathogenic fibrils in diseases. In addition to the glycine-rich region, the truncated RRM2, but not the intact RRM2, plays a key role in forming cytoplasmic inclusions in neuronal cells. Our data thus suggest that the process that disrupts the dimeric structure, such as the proteolytic cleavage of TDP-43 within the RRM2 that removes the N-terminal dimerization domain, may produce unassembled truncated RRM2 fragments with abnormally exposed ß-strands, which can oligomerize into high-order inclusions.

Minimizing nitrous oxide in biological nutrient removal from municipal wastewater by controlling copper ion concentrations.

In this study, nitrous oxide (N(2)O) production during biological nutrient removal (BNR) from municipal wastewater was reported to be remarkably reduced by controlling copper ion (Cu(2+)) concentration. Firstly, it was observed that the addition of Cu(2+) (10-100 µg/L) reduced N(2)O generation by 54.5-73.2 % and improved total nitrogen removal when synthetic wastewater was treated in an anaerobic-aerobic (with low dissolved oxygen) BNR process. Then, the roles of Cu(2+) were investigated. The activities of nitrite and nitrous oxide reductases were increased by Cu(2+) addition, which accelerated the bio-reductions of both nitrite to nitric oxide (NO (2) (-) → NO) and nitrous oxide to nitrogen gas (N(2)O → N(2)). The quantitative real-time polymerase chain reaction assay indicated that Cu(2+) addition increased the number of N(2)O reducing denitrifiers. Further investigation showed that more polyhydoxyalkanoates were utilized in the Cu(2+)-added system for denitrification. Finally, the feasibility of reducing N(2)O generation by controlling Cu(2+) was examined in two other BNR processes treating real municipal wastewater. As the Cu(2+) in municipal wastewater is usually below 10 µg/L, according to this study, the supplement of influent Cu(2+) to a concentration of 10-100 µg/L is beneficial to reduce N(2)O emission and improve nitrogen removal when sludge concentration in the BNR system is around 3,200 mg/L.

Biological nutrient removal by applying modified four step-feed technology to treat weak wastewater.

For weak municipal wastewater (COD ≤ 200 mg L(-1), NH(4)(+)-N ≤ 40 mg L(-1)) with low influent C/N, a pilot modified four step-feed process was applied for simultaneous biological nitrogen (N) and phosphorus (P) removal under different inflow distribution ratios. It was designed with a short hydraulic retention time of 8.7h to raise influent load, and the optimal effluent performance of COD, NH(4)(+)-N, total nitrogen (TN) and total phosphorus (TP) were 33.05, 0.58, 9.26 and 0.46 mg L(-1), respectively with inflow distribution ratio of 20:35:35:10%. More than 74% of carbon sources were utilized effectively for phosphorus release and denitrification. About 16.7% of TN was removed through simultaneous nitrification and denitrification in oxic zones. Moreover, the commendable sludge settling with a sludge loading of 0.04-0.1 kg COD/kg MLSS d, attributed to the higher mixed liquor suspended solids (MLSS) and the alternating anoxic/oxic operational mode. In addition, the pre-anoxic zone designed was beneficial for both nitrate reduction and anaerobic phosphorus release.