Temporal Dynamics and Performance Association of the Tetrasphaera-Enriched Microbiome for Enhanced Biological Phosphorus Removal

Tetrasphaera have been recently identified based on the 16S ribosomal RNA(rRNA)gene as among the most abundant polyphosphate-accumulating organisms(PAOs)in global full-scale wastewater treatment plants(WWTPs)with enhanced biological phosphorus removal(EBPR).However,it is unclear how Tetrasphaera PAOs are selectively enriched in the context of the EBPR microbiome.In this study,an EBPR microbiome enriched with Tetrasphaera(accounting for 40%of 16S sequences on day 113)was built using a top-down design approach featuring multicarbon sources and a low dosage of allylthiourea.The microbiome showed enhanced nutrient removal(phosphorus removal~85%and nitrogen removal~80%)and increased phosphorus recovery(up to 23.2 times)compared with the seeding activated sludge from a local full-scale WWTP.The supply of 1 mg·L^(-1)allylthiourea promoted the coselection of Tetrasphaera PAOs and Microlunatus PAOs and sharply reduced the relative abundance of both ammonia oxidizer Nitrosomonas and putative competitors Brevundimonas and Paracoccus,facilitating the establishment of the EBPR microbiome.Based on 16S rRNA gene analysis,a putative novel PAO species,EBPR-ASV0001,was identified with Tetrasphaera japonica as its closest relative.This study provides new knowledge on the establishment of a Tetrasphaera-enriched microbiome facilitated by allylthiourea,which can be further exploited to guide future process upgrading and optimization to achieve and/or enhance simultaneous biological phosphorus and nitrogen removal from high-strength wastewater.

Screening of Polyphosphate Accumulating Organisms and Their Phosphorus Removal Performance

[Objectives]To study the phosphorus removal performance of phosphate accumulating organisms(PAOs).[Methods]Activated sludge from domestic sewage treatment plant was used as the strain source,and phosphate accumulating organisms were screened by plate streaking method and dilution coating plate method.Six kinds of excellent phosphate accumulating organisms were obtained by metachromatic granule staining experiment,total phosphorus experiment and simulated sewage phosphorus removal experiment to assist the observation of bac-terial morphology and experiment of phosphorus removal capacity.In addition,the influencing factors of phosphorus removal capacity(nitrogen source,trace metal ions)were analyzed.[Results]In the case of simulated sewage,the phosphorus removal rate of strain b was the highest,reaching 66.25%,while the phosphorus removal rate of strain e and f was about 10%lower than that of the phosphorus uptake experiment.[Conclusions]This study is expected to provide a theoretical reference for the gradual optimization of the screening method of phosphorus re-moval bacteria in domestic sewage treatment.

Comparing results of cultured and uncultured biological methods used in biological phosphorus removal

Increasing attention has been paid to phosphate-accumulating organisms （PAOs） for their important role in biological phosphorus removal. In this study, microbial communities of PAOs cultivated under different carbon sources （sewage, glucose, and sodium acetate） were investigated and compared through culture-dependent and culture-independent methods, respectively. The results obtained using denaturing gradient gel electrophoresis （DGGE） of polymerase chain reaction-amplified 16S rDNA fragments revealed that the diversity of bacteria in a sewage-fed reactor （1#） was much higher than in a glucose-fed one （2#） and a sodium acetate-fed one （3#）; there were common PAOs in three reactors fed by different carbon sources. Five strains were separated from three systems by using a phosphaterich medium; they were from common bacteria isolated and three isolates could not be found in DGGE profile at all. Two isolates had good phosphorus removal ability. When the microbial diversity was studied, the molecular biological method was better than the culture-dependent one. When phosphorus removal characteristics were investigated, culture-dependent approach was more effective. Thus a combination of two methods is necessary to have a comprehensive view of PAOs.

Effects of COD to Phosphorus Ratios on the Metabolism of PAOs in Enhanced Biological Phosphorus Removal with Different Carbon Sources

To elucidate the phosphorus removal and metabolism under various COD / P ratio,a sludge highly enriched in PAOs was used to investigate the impacts of COD / P in batch tests under different carbon supply conditions. Acetate,propionate and a mixture of acetate and propionate at a ratio of 3 ∶ 1( COD basis) was used as carbon sources with the COD / P of 20,15,10 and 5. 0 g COD /gP,respectively. The minimum COD / P ratios for complete P removal were found to be 8. 24 g COD /gP for acetate,11. 40 g COD /gP for propionate and9. 10 g COD /gP for the 3 ∶ 1 mixture of acetate and propionate. Converted to a mass basis,all three cases had a very similar ratio of 7. 7 g VFA /gP,which represented a useful guide for operation of EBPR plants to identify possible shortages in VFAs. The trend in PHV accumulation during the anaerobic period along with the decrease of COD / P ratios suggested that,PAOs may use the TCA pathway for anaerobic VFA uptake to maintain the required NADH production with reduced glycogen degradation. During the aerobic phase,the glycogen pool was reduced but remained enough compared to the requirement for anaerobic VFA uptake,and the synthesis and degradation of glycogen was not the inhibition factor of PAOs.

Role of extracellular exopolymers on biological phosphorus removal

Three sequencing batch reactors supplied with different carbon sources were investigated. The system supplied with glucose gained the best enhanced biological phosphorus removal although all of the three reactors were seeded from the same sludge. With the measurement of poly-β-hydroxyalkanoate （PHA） concentration, phosphorus content in sludge and extracellular exopolymers （EPS） with scanning electron microscopy （SEM） combined with energy dispersive spectrometry （EDS）, it was found that the biosorption effect of EPS played an important role in phosphorus removal and that the amount of PHA at the end of anaerobic phase was not the only key factor to determine the following phosphorus removal efficiency.

Enhanced Nutrient Removal with Upflow Biological Aerated Filter for Reclaimed Water

A two-stage upflow biological aerated filter was designed as an advanced treatment process to optimize the operating parameters and study the correlative factors influencing the efficiency of nitrification, denitrification and phosphorus removal. The experimental results showed that the final effluent of the two-stage upflow biofilter process operated in series could meet the stringent limits of the reclaimed water for the total nitrogen of 2 mg/L, and total phosphorus of 0.3 mg/L. The high treatment efficiency allowed the reactor operating at very high hydraulic loadings and reaching nearly complete nitrification and denitrifieation.

Nitrogen and phosphorus removal in pilot-scale anaerobic-anoxic oxidation ditch system

To achieve high efficiency of nitrogen and phosphorus removal and to investigate the rule of simultaneous nitrification and denitrification phosphorus removal （SNDPR）, a whole course of SNDPR damage and recovery was studied in a pilot-scale, anaerobicanoxic oxidation ditch （OD）, where the volumes of anaerobic zone, anoxic zone, and ditches zone of the OD system were 7, 21, and 280 L, respectively. The reactor was fed with municipal wastewater with a flow rate of 336 L/d. The concept of simultaneous nitrification and denitrification （SND） rate （rSND） was put forward to quantify SND. The results indicate that： （1） high nitrogen and phosphorus removal efficiencies were achieved during the stable SND phase, total nitrogen （TN） and total phosphate （TP） removal rates were 80% and 85%, respectively; （2） when the system was aerated excessively, the stability of SND was damaged, and rSND dropped from 80% to 20% or less; （3） the natural logarithm of the ratio of NOx to NH4^＋ in the effluent had a linear correlation to oxidation-reduction potential （ORP）; （4） when NO3^- was less than 6 mg/L, high phosphorus removal efficiency could be achieved; （5） denitrifying phosphorus removal （DNPR） could take place in the anaerobic-anoxic OD system. The major innovation was that the SND rate was devised and quantified.

Denitrifying phosphorus removal in a step-feed CAST with alternating anoxic-oxic operational strategy

A bench-scale cyclic activated sludge technology （CAST） was operated to study the biological phosphorus removal performance and a series of batch tests was carried out to demonstrate the accumulation of denitrifying polyphosphate-accumulating organisms （DNPAOs） in CAST system. Under all operating conditions, step-feed CAST with enough carbon sources in influent had the highest nitrogen and phosphorus removal efficiency as well as good sludge settling performance. The average removal rate of COD, NH4^＋-N, PO4^3--P and total nitrogen （TN） was 88.2%, 98.7%, 97.5% and 92.1%, respectively. The average sludge volume index （SVI） was 133 mL/g. The optimum anaerobic/aerobic/anoxic （AOA） conditions for the cultivation of DNPAOs could be achieved by alternating anoxic/oxic operational strategy, thus a significant denitrifying phosphorus removal occurred in step-feed CAST. The denitrification of NO^x--N completed quickly due to step-feed operation and enough carbon sources, which could enhance phosphorus release and further phosphorus uptake capability of the system. Batch tests also proved that polyphosphate-accumulating organisms （PAOs） in the step-feed process had strong denitrifying phosphorus removal capacity. Both nitrate and nitrite could be used as electron acceptors in denitrifying phosphorus removal. Low COD supply with step-feed operation strategy would favor DNPAOs accumulation.

Removal of nitrogen and phosphorus in a combined A^2/O-BAF system with a short aerobic SRT

A bench-scale anaerobic/anoxic/aerobic process-biological aerated filter （A^2/O-BAF） combined system was carded out to treat wastewater with lower C/N and C/P ratios. The A^2/O process was operated in a short aerobic sludge retention time （SRT） for organic pollutants and phosphorus removal, and denitrification. The subsequent BAF process was mainly used for nitrification. The BAF effluent was partially returned to anoxic zone of the A^2/O process to provide electron acceptors for denitrification and anoxic P uptake. This unique system formed an environment for reproducing the denitdfying phosphate-accumulating organisms （DPAOs）. The ratio of DPAOs to phosphorus accumulating organisms （PAOs） could be maintained at 28% by optimizing the organic loads in the anaerobic zone and the nitrate loads into the anoxic zone in the A^2/O process. The aerobic phosphorus over-uptake and discharge of excess activated sludge was the main mechanism of phosphorus removal in the combined system. The aerobic SRT of the A^2/O process should meet the demands for the development of aerobic PAOs and the restraint on the nitrifiers growth, and the contact time in the aerobic zone of the A^2/O process should be longer than 30 min, which ensured efficient phosphorus removal in the combined system. The adequate BAF effluent return rates should be controlled with 1--4 mg/L nitrate nitrogen in the anoxic zone effluent of A^2/O process to achieve the optimal nitrogen and phosphorus removal efficiencies.

Effect of carbon source and nitrate concentration on denitrifying phosphorus removal by DPB sludge

Effect of added carbon source and nitrate concentration on the denitrifying phosphorus removal by DPB sludge was systematically studied using batch experiments, at the same time the variation of ORP was investigated. Results showed that the denitrifying and phosphorus uptake rate in anoxic phase increased with the high initial anaerobic carbon source addition. However once the initial COD concentration reached a certain level, which was in excess to the PHB saturation of poly-P bacteria, residual COD carried over to anoxic phase inhibited the subsequent denitrifying phosphorus uptake. Simultaneously, phosphate uptake continued until all nitrate was removed, following a slow endogenous release of phosphate. High nitrate concentration in anoxic phase increased the initial denitrifying phosphorus rate. Once the nitrate was exhausted, phosphate uptake changed to release. Moreover, the time of this turning point occurred later with the higher nitrate addition. On the other hand, through on-line monitoring the variation of the ORP with different initial COD concentration, it was found ORP could be used as a control parameter for phosphorus release, but it is impossible to utilize ORP for controlling the denitrificaion and anoxic phosphorus uptake operations.

Characteristics of anoxic phosphors removal in sequence batch reactor

The characteristics of anaerobic phosphorus release and anoxic phosphorus uptake were investigated in sequencing batch reactors using denitrifying phosphorus removing bacteria （DPB） sludge. The lab-scale experiments were accomplished under conditions of various nitrite concentrations （5.5, 9.5, and 15 mg/L） and mixed liquor suspended solids （MLSS） （1844, 3231, and 6730 mg/L）. The results obtained confirmed that nitrite, MLSS, and pH were key factors, which had a significant impact on anaerobic phosphorus release and anoxic phosphorus uptake in the biological phosphorous removal process. The nitrites were able to successfully act as electron acceptors for phosphorous uptake at a limited concentration between 5.5 and 9.5 mg/L. The denitrification and dephosphorous were inhibited when the nitrite concentration reached 15 mg/L. This observation indicated that the nitrite would not inhibit phosphorus uptake before it exceeded a threshold concentration. It was assumed that an increase of MLSS concentration from 1844 mg/L to 6730 mg/L led to the increase of denitrification and anoxic P-uptake rate. On the contrary, the average P-uptake/N denitrifying reduced from 2.10 to 1.57 mg PO4^3--P/mg NO3^--N. Therefore, it could be concluded that increasing MLSS of the DEPHANOX system might shorten the reaction time of phosphorus release and anoxic phosphorus uptake. However, excessive MLSS might reduce the specific denitrifying rate. Meanwhile, a rapid pH increase occurred at the beginning of the anoxic conditions as a result of denitrification and anoxic phosphate uptake. Anaerobic P release rate increased with an increase in pH. Moreover, when pH exceeded a relatively high value of 8.0, the dissolved P concentration decreased in the liquid phase, because of chemical precipitation. This observation suggested that pH should be strictly controlled below 8.0 to avoid chemical precipitation if the biological denitrifying phosphorus removal capability is to be studied accurately.

Operation of three parallel AN/AO processes to enrich denitrifying phosphorus removing bacteria for low strength wastewater treatment

Three parallel anaerobic-anoxic/anaerobic-aerobic （AN/AO） processes were developed to enrich denitrifying phosphorus removal bacteria （DPB） for low strength wastewater treatment. The main body of the parallel AN/AO process consists of an AN （anaerobic-anoxic） process and an AO （anaerobic-aerobic） process. In the AO process, the common phosphorus accumulating organisms （PAOs） was dominate, while in the AN process, DPB was dominate, The volume of anaerobic zone（Vana）：anoxie zone（Vano） ： aerobic zone （Vaer） for the parallel AN/AO process is 1：1：1 in contrast with a Vana：Vaer and Vano：Vaer of 1：2 and 1：4 for a traditional biological nutrient removal process （BNR）. Process 3 excels in the 3 processes on the basis of COD, TN and TP removal. For 4 month operation, the effluent COD concentration of process 3 did not exceed 60 mg/L; the effluent TN concentration of process 3 was lower than 15 mg/L; and the effluent TP concentration of process 3 was lower than 1 mg/L.

Enhanced biological phosphorus removal using thermal alkaline hydrolyzed municipal wastewater biosolids

This study reports the feasibility of using municipal wastewater biosolids as an alternative carbon source for biological phosphorus removal.The biosolids were treated by a lowtemperature,thermal alkaline hydrolysis process patented by Lystek International Inc.(Cambridge,ON,Canada)to produce short-chain volatile fatty acids and other readily biodegradable organics.Two sequencing batch reactors(SBRs)were operated with synthetic volatile fatty acids(Syn VFA)and readily biodegradable organics produced from the alkaline hydrolysis of municipal wastewater biosolids(Lystek)as the carbon source,respectively.Municipal wastewaters with different strengths and COD:N:P ratios were tested in the study.The reactors’performances were compared with respect to nitrogen and phosphorus removal.It was observed that phosphorus removal efficiencies were between 98%–99%and 90%–97%and nitrogen removal efficiencies were 78%–81%,and 67%for the Syn VFA and Lystek,respectively.However,the kinetics for phosphorus release and uptake during the anaerobic and aerobic stages with Lystek were observed to be significantly lower than Syn VFA due to the presence of higher order VFAs(C4 and above)and other fermentable organics in the Lystek.

Efects of idle time on biological phosphorus removal by sequencing batch reactors

Three identical sequencing batch reactors （SBRs） were operated to investigate the effects of various idle times on the biological phosphorus （P） removal. The idle times were set to 3 hr （R1）, 10 hr （R2） and 17 hr （R3）. The results showed that the idle time of a SBR had potential impact on biological phosphorus removal, especially when the influent phosphorus concentration increased. The phosphorus removal efficiencies of the R2 and R3 systems declined dramatically compared with the stable R1 system, and the Prelease and P-uptake rates of the R3 system in particular decreased dramatically. The PCR-DGGE analysis showed that uncultured Pseudomonas sp. （GQ183242.1） and β-Proteobacteria （AY823971） were the dominant phosphorus removal bacteria for the R1 and R2 systems, while uncultured γ-Proteobacteria were the dominant phosphorus removal bacteria for the R3 system. Glycogen-accumulating organisms （GAOs）, such as uncultured Sphingomonas sp. （AM889077）, were found in the R2 and R3 systems. Overall, the R1 system was the most stable and exhibited the best phosphorus removal efficiency. It was found that although the idle time can be prolonged to allow the formation of intracellular polymers when the phosphorus concentration of the influent is low, systems with a long idle time can become unstable when the influent phosphorus concentration is increased.

Kinetic analysis of anaerobic phosphorus release during biological phosphorus removal process

Enhanced biological phosphorus removal(EBPR)is a commonly used and sustainable method for phosphorus removal from wastewater.Poly-β-hydroxybutyrate(PHB),polyphosphate,and glycogen are three kinds of intracellular storage polymers in phosphorus accumulation organisms.The variation of these polymers under different conditions has an apparent influence on anaerobic phosphorus release,which is very important for controlling the performance of EBPR.To obtain the mechanism and kinetic character of anaerobic phosphorus release,a series of batch experiments were performed using the excessively aerated sludge from the aerobic unit of the biological phosphorus removal system in this study.The results showed that the volatile suspended solid(VSS)had an increasing trend,while the mixed liquid suspended sludge(MLSS)and ashes were reduced during the anaerobic phosphorus release process.The interruption of anaerobic HAc-uptake and phosphorus-release occurs when the glycogen in the phosphorus-accumulating-organisms is exhausted.Under the condition of lower initial HAc-COD,HAc became the limiting factor after some time for anaerobic HAc uptake.Under the condition of higher initial HAc-COD,HAc uptake was stopped because of the depletion of glyco-gen in the microorganisms.The mean ratio ofΔ_(ρP)/Δ_(ρPHB),Δ_(ρ)GLY/ΔρPHB,Δ_(ρP)/ΔCOD,andΔ_(ρPHB)/ΔCOD was 0.48,0.50,0.44,and 0.92,respectively,which was nearly the same as the theoretical value.The calibrated kinetic parameters of the HAc-uptake and phosphorus-release model were evaluated as follows:QHAc,max was 164 mg/(g·h),QP,max was 69.9 mg/(g·h),Kgly was 0.005,and KCOD was 3 mg/L.An apparently linear correlation was observed between the ratio ofΔ_(ρP)/ΔCOD and pH of the solution,and the equation between them was obtained in this study.Enhanced biological phosphorus removal(EBPR)is a commonly used and sustainable method for phosphorus removal from wastewater.Poly-β-hydroxybutyrate(PHB),polyphosphate,and glycogen are three kinds of intracellular storage polymers in phosphorus accumulation organisms.The variation of these polymers under different conditions has an apparent influence on anaerobic phosphorus release,which is very important for controlling the performance of EBPR.To obtain the mechanism and kinetic character of anaerobic phosphorus release,a series of batch experiments were performed using the excessively aerated sludge from the aerobic unit of the biological phosphorus removal system in this study.

Research on polyhydroxyalkanoates and glycogen transformations: Key aspects to biologic nitrogen and phosphorus removal in low dissolved oxygen systems

In this paper,a study was conducted on the effect of polyhydroxyalkanoates(PHA)and glycogen transformations on biologic nitrogen and phosphorus removal in low dissolved oxygen(DO)systems.Two laboratory-scale sequencing batch reactors(SBR1 and SBR2)were operating with anaerobic/aerobic(low DO,0.15-0.45 mg·L^(-1))configurations,which cultured a propionic to acetic acid ratio(molar carbon ratio)of 1.0 and 2.0,respectively.Fewer poly-3-hydroxybutyrate(PHB),total PHA,and glycogen transformations were observed with the increase of propionic/acetic acid,along with more poly-3-hydroxyvalerate(PHV)and poly-3-hydroxy-2-methyvalerate(PH2MV)shifts.The total nitrogen(TN)removal efficiency was 68%and 82%in SBR1 and SBR2,respectively.In the two SBRs,the soluble ortho-phosphate(SOP)removal efficiency was 94%and 99%,and the average sludge polyphosphate(poly-P)content(g·g-MLVSS^(-1))was 8.3%and 10.2%,respectively.Thus,the propionic to acetic acid ratio of the influent greatly influenced the PHA form and quantity,glycogen transformation,and poly-P contained in activated sludge and further determined TN and SOP removal efficiency.Moreover,significant correlations between the SOP removal rate and the(PHV+PH2MV)/PHA ratio were observed(R^(2)>0.99).Accordingly,PHA and glycogen transformations should be taken into account as key components for optimizing anaerobic/aerobic(low DO)biologic nitrogen and phosphorus removal systems.

Characteristics of intracellular polyphosphate granules and phosphorus-absorption of a marine polyphosphateaccumulating bacterium, Halomonas sp. YSR-3

Halomonas sp.YSR-3 was isolated from the Yellow Sea and identified as a polyphosphate-accumulating bacterium and the characteristics of its intracellular polyphosphate(polyP)granules and phosphorus absorption were studied.Most YSR-3 cells stored one or two polyP granules in regular appearance and high-density.The diameter of the granules was about 400 nm measuring by a transmission electron microscope(TEM).After stained with 4,6-diamidino-2-phenylindole(DAPI)and visualized by a fluorescence microscope,the cells turned blue and the granules were bright yellow.The composition of granules includes P(major ingredient),Mg,S,K,and Ca as detected by an energy dispersive X-ray spectrometer(EDS).When inorganic phosphorus(po34-)and ferric ion(Fe3+)were added into media,the biomass increased and the cells formed intracellular polyP granules owing to the phosphorus assimilation from media.The YSR-3 obtained higher biomass by adding 0.02 g/L FePO4 than 0.005 g/L and 0.01 g/L FePO4;however,the phosphorus absorption was higher with 0.01 g/L FePO4 than 0.005 g/L and 0.02 g/L FePO4.The optical density at wavelength 480 nm(OD480nm)was 0.79 and 100%cells could form intracellular polyP granules.These results show that strain YSR-3 is able to acquire higher biomass and absorb more inorganic phosphorus when 0.01 g/L FePO4 is added.The characteristics of absorbing and storing phosphorus as intracellular inorganic polyP granules have a potential for application in high-efficiency phosphorus removal in wastewater treatment.

Influence of Azo Dye on Metabolism of Phosphorus Accumulating Organisms Linked to Transformation of Intracellular Storage Products

In this study, the influence of azo dye of methyl red (MR) on COD, dye and phosphorus removal and the transformation of polyhydroxyalkanoate (PHA) and glycogen of phosphate accumulating organisms in enhanced biological phosphorus removal (EBPR) system were investigated. The results indicated COD and dye removal efficiencies were decreased from 97.9% to 72.8% and 99.7% to 82.0%, respectively, when MR concentration was increased from 0 to 40 mg/L. Low MR concentration (5 mg/L) had no influence on P removal and transformation of PHA and glycogen. However, P removal, PHA production and consumption, and glycogen replenishment were seriously inhibited at high MR concentration, while glycogen hydrolysis was simulated at MR concentration of 20 and 40 mg/L. The transformations of PHA and glycogen at aerobic condition were more sensitive to those at anaerobic condition at high MR concentration. These results demonstrated dye and its intermediate products would inhibit the metabolism of polyphosphate accumulating organisms, which should be taken into account in future work.

Nitrous oxide emission by denitrifying phosphorus removal culture using polyhydroxyalkanoates as carbon source

Nitrous oxide （N2O） emission has been reported to be enhanced during denitrification when internally-stored compounds are used as carbon sources. However, negligible N2O emissions have been detected in the few studies where polyhydroxyalkanoates （PHA） were specifically used. This study investigated and compared the potential enhancement of N2O production, based on utilization of an internally-stored polymer and external carbon （acetate） by a denitrifying phosphorus removal culture. Results indicated that at relatively low chemical oxygen demand-to-nitrogen （COD/N） ratios, more nitrite was reduced to N2O in the presence of an external carbon source as compared to an internal carbon source （PHA）. At relatively higher COD/N ratios, similar N2O reduction rates were obtained in all cases regardless of the type of carbon source available. N2O reduction rates were, however, generally higher in the presence of an internal carbon source. Results from the study imply that when the presence of an external carbon source is not sufficient to support denitrification, it is likely competitively utilized by different metabolic pathways of denitrifying polyphosphate accumulating organisms （DPAOs） and other ordinary denitfifiers. This study also reveals that the consumption of PHA is potentially the rate-limiting step for N2O reduction during denitrification.

Advanced nitrogen and phosphorus removal in A^(2)O-BAF system treating low carbon-to-nitrogen ratio domestic wastewater

A laboratory-scale anaerobic-anoxic-aerobic process(A^(2)O)with a small aerobic zone and a bigger anoxic zone and biologic aerated filter(A^(2)O-BAF)system was operated to treat low carbon-to-nitrogen ratio domestic wastewater.The A^(2)O process was employed mainly for organic matter and phosphorus removal,and for denitrification.The BAF was only used for nitrification which coupled with a settling tank Compared with a conventional A^(2)O process,the suspended activated sludge in this A^(2)OBAF process contained small quantities of nitrifier,but nitrification overwhelmingly conducted in BAF.So the system successfully avoided the contradiction in sludge retention time(SRT)between nitrifying bacteria and phosphorus accumulating organisms(PAOs).Denitrifying phosphorus accumulating organisms(DPAOs)played an important role in removing up to 91%of phosphorus along with nitrogen,which indicated that the suspended activated sludge process presented a good denitrifying phosphorus removal performance.The average removal efficiency of chemical oxygen demand(COD),total nitrogen(TN),total phosphorus(TP),and NH_(4)^(+)-N were 85.56%,92.07%,81.24%and 98.7%respectively.The effluent quality consistently satisfied the national first level A effluent discharge standard of China.The average sludge volume index(SVI)was 85.4 mL·g^(-1)additionally,the volume ratio of anaerobic,anoxic and aerobic zone in A^(2)O process was also investigated,and the results demonstrated that the optimum value was 1:6:2.