Enhanced Biological Phosphorus Removal with Pseudomonas putida GM6 from Activated Sludge

The enhanced biological phosphorus removal （EBPR） method is widely adopted for phosphorus removal from wastewater, yet little is known about its microbiological and molecular mechanisms. Therefore, it is difficult to predict and control the deterioration of the EBPR process in a large-scale municipal sewage treatment plant. This study used a novel strain isolated in the laboratory, Pseudomonas putida GM6, which had a high phosphate accumulating ability and could recover rapidly from the deteriorated system and enhance the capability of phosphorus removal in activated sludge. Strain GM6 marked with gfp gene, which was called GMTR, was delivered into a bench-scale sequencing batch reactor （SBR） of low efficiency, to investigate the colonization of GMTR and removal of phosphorus. After 21 days, the proportion of GMTR in the total bacteria of the sludge reached 9.2%, whereas the phosphorus removal rate was 96%, with an effluent concentration of about 0.2 mg L^-1. In the reactor with the addition of GMTR, phosphorus was removed quickly, in 1 h under anaerobic conditions, and in 2 h under aerobic conditions. These evidences were characteristic of EBPR processes. Field testing was conducted at a hospital sewage treatment facility with low phosphorus removal capability. Twentyone days after Pseudomonas putida GM6 was added, effluent phosphorus concentration remained around 0.3 mg L^-1, corresponding to a removal rate of 96.8%. It was therefore demonstrated that Pseudomonas putida GM6 could be used for a quick startup and enhancement of wastewater biological phosphorus removal, which provided a scientific basis for potential large-scale engineering application.

Enhanced biological nutrient removal in modified carbon source division anaerobic anoxic oxic process with return activated sludge pre-concentration

A pilot-scale modified carbon source division anaerobic anoxic oxic(AAO) process with pre-concentration of returned activated sludge(RAS) was proposed in this study for the enhanced biological nutrient removal(BNR) of municipal wastewater with limited carbon source. The influent carbon source was fed in step while a novel RAS pre-concentration tank was adopted to improve BNR efficiency, and the effects of an influent carbon source distribution ratio and a RAS pre-concentration ratio were investigated. The results show that the removal efficiency of TN is mainly influenced by the carbon source distribution ratio while the TP removal relies on the RAS pre-concentration ratio. The optimum carbon source distribution ratio and RAS pre-concentration ratio are 60% and 50%, respectively, with an inner recycling ratio of 100% under the optimum steady operation of pilot test, reaching an average effluent TN concentration of 9.8 mg·L-1with a removal efficiency of 63% and an average TP removal efficiency of 94%. The mechanism of nutrient removal is discussed and the kinetics is analyzed. The results reveal that the optimal carbon source distribution ratio provides sufficient denitrifying carbon source to each anoxic phase, reducing nitrate accumulation while the RAS pre-concentration ratio improves the condition of anaerobic zone to ensure the phosphorus release due to less nitrate in the returned sludge. Therefore, nitrifying bacteria, denitrifying bacteria and phosphorus accumulation organisms play an important role under the optimum condition, enhancing the performance of nutrient removal in this test.

Effect of Return Sludge Pre-concentration on Biological Phosphorus Removal in a Novel Oxidation Ditch

A pilot-scale,pre-anoxic-anaerobic oxidation ditch was used in this study to treat municipal wastewater with limited carbon source.A novel return activated sludge(RAS) pre-concentration tank was adopted for improv-ing the phosphorus removal efficiency and the effects of RAS pre-concentration ratio were studied.Under the opti-mal operational condition,the suspended total phosphorus(STP) and the total phosphorus(TP) removal efficiencies were around 58.9% and 63.9% respectively and the effluent-P was lower than 0.8 mg·L-1.The reason is that with the optimal RAS pre-concentration ratio,nitrate is completely removed with endogenous carbon source and the secondary phosphorus release is strictly restrained in the pre-anoxic tank.Therefore,the anaerobic phosphorus release and the carbon source uptake by phosphorus accumulation organisms(PAOs) in the sludge,which are ex-tremely important to the phosphorus removal process,can be fully satisfied.Furthermore,the oxidation-reduction potential is proved to be suitable for controlling the RAS pre-concentration ratio due to influent fluctuation and varied conditions.The novel modified system is also beneficial for PAO accumulation.

Optimum operation conditions of nitrogen and phosphorus removal by a biofilm-activated-sludge system

In the biofilm and activated sludge combined system, denitrifying bacteria attached on the fibrous carriers in the anoxic tank, while the sludge containing nitrifying and phosphorus removal bacteria was only recirculated between the aerobic and anaerobic tanks. Therefore, the factors affected and restricted nitrification, denitrification and phosphorus removal in a traditional A/A/O process were resolved. This paper describes the optimum operation conditions for nitrogen and phosphorus removal using this system.

Effect of Ferric Chloride on the Properties of Biological Sludge in Co-precipitation Phosphorus Removal Process

This paper studied the effect of ferric chloride on waste sludge digestion,dewatering and sedimentation under the optimized doses in co-precipitation phosphorus removal process.The experimental results showed that the concentration of mixed liquid suspended solid(MLSS) was 2436 mg.L-1 and 2385 mg.L-1 in co-precipitation phosphorus removal process(CPR) and biological phosphorous removal process(BPR),respectively.The sludge reduction ratio for each process was 22.6% and 24.6% in aerobic digestion,and 27.6% and 29.9% in anaerobic digestion,respectively.Due to the addition of chemical to the end of aeration tank,the sludge content of CPR was slightly higher than that of BPR,but the sludge reduction rate for both processes had no distinct difference.The sludge volume index(SVI) and sludge specific resistance of BPR were 126 ml.g-1 and 11.7×1012 m.kg-1,respectively,while those of CPR were only 98 ml.g-1 and 7.1×1012 m.kg-1,indicating that CPR chemical could improve sludge settling and dewatering.

Comparative study of two biological nitrogen removal processes:A/O process and step-feeding process

Two biological nitrogen removal processes are compared in the aspect of nitrogen removal, process operation and energy saving. Results show that when the returned sludge ratio is 50% of the inflow rate, the step-feeding process achieves over 80% total nitrogen （TN） removal efficiency, but the TN removal efficiency of the A/O process is only 40%. Moreover, filamentous sludge bulking can be well restrained in the step-feeding process. Given the conditions of a returned sludge ratio of 100% and a nitrifying liquor recycle ratio of 200%, the TN removal efficiency is 78.32% in the A/O process, but the sludge volume index （SVI） value increases to 143 mL/g. In the step-feeding process, the SVI is only 94.4 mL/g when the TN removal efficiency reaches 81. 1%. The step-feeding process has distinct advantages over the A/O process in the aspects of practicability, nitrogen removal and operating stability.

Performance of biological phosphorus removal and characteristics of microbial community in the oxic-settling-anaerobic process by FISH analysis

Performance of biological phosphorus removal in the oxic-settling-anaerobic(OSA) process was investigated. Cell staining and fluorescent in situ hybridization(FISH) were used to analyze characteristics and microbial community of sludge. Experimental results showed that phosphorus removal efficiency was near 60% and the amount of biological phosphorus accumulation in aerobic sludge of the OSA system was up to 26.9 mg/g. Biological phosphorus removal efficiency was partially inhibited by carbon sources in the continuous OSA system. Contrasted to the OSA system,biological phosphorus removal efficiency was enhanced by 14% and the average total phosphorus(TP) contents of aerobic sludge were increased by 0.36 mg/g when sufficient carbon sources were supplied in batch experiments. Staining methods indicated that about 35% of microorganisms had typical characteristics of phosphorus accumulating organisms(PAOs) . FISH analysis demonstrated that PAOMIX-binding bacteria were predominant microbial communities in the OSA system,which accounted for around 28% of total bacteria.

Community structures and population dynamics of “Candidatus Accumulibacter” in activated sludges of wastewater treatment plants using ppk1 as phylogenetic marker

Candidatus Accumulibacter has been identified as dominant polyphosphate-accumulating organisms（PAOs） in enhanced biological phosphorus（P） removal（EBPR） from wastewater.This study revealed the relevance of community structure, abundance and seasonal population dynamics of Candidatus Accumulibacter to process operation of wastewater treatment plants（WWTPs） in China using ppk1 gene as phylogenetic marker. All sludge samples had properties of denitrifying P removal using nitrate as an electron acceptor.Accumulibacter abundance in the anaerobic-anoxic-oxic（A^2O） process was the highest（26%of total bacteria）, and higher in winter than in summer with a better EBPR performance.Type-II was the dominant Accumulibacter in all processes, and type-I accounted for a small proportion of total Accumulibacter. The abundance of Clade-IIC as the most dominant clade reached 2.59 × 10~9 cells/g MLSS and accounted for 87.3% of total Accumulibacter. Clade IIC mainly contributed to denitrifying P removal. Clades IIA, IIC and IID were found in all processes, while clade-IIF was only found in oxidation ditch process through phylogenetic analysis. High proportion of clade IID to total Accumulibacter led to poor performance of aerobic P-uptake in inverted A^2O process. Therefore, Accumulibacter clades in WWTPs were diverse, and EBPR performance was closely related to the clade-level community structures and abundances of Accumulibacter.

Effects of low temperature on sludge settleability and nutrients removal performance treating domestic wastewater

In order to investigate the effect of temperature on activated sludge systems,sludge settleability,nitrogen and phosphorus removal processes were investigated by changing temperature variation patterns using 4 sequencing batch reactors (SBR).The results showed that no matter temperature changes in the range of 15-22 ℃ (decrease or increase) gradually or sharply,it has little effect on nitrogen and phosphorus removal processes.But when temperature decreases to 12 ℃,biochemical reactions will be inhibitted obviously.At least 1 sludge retention time (SRT) is needed for nitrification adapt to new temperature envirionment,and more time is necessary for phosphorus removal process.When temperature increases from 12 ℃ to 22 ℃ sharply,nitrification process deteriorates seriously,but phosphorus removal process shows no change.In addition,sludge settleability deteriorates when temperature changes sharply (decrease or increase),but the reasons are different.Under temperature decrease condition,it is mainly caused by the increase of accumulated extracellular polymeric substances (EPS).Under temperature increase condition,the loosing sludge flocs' configuration is the main reason.It should be pointed out that the filamentous bacteria content during the entire experiment keeps almost constant,and the sludge settleability variations are the reflection of the change of sludge physicochemical characteristics.

Modification of the activated sludge model for chemical dosage

Full-scale experiments have been carried out to adapt the activated sludge model ASM2d to include the influence of metal dosage （Fe^3＋ and Al^3＋） for phosphorus removal. Phosphorus removal rates, nitrification rates, as well as pH and sludge settling performance, were evaluated as functions of the metal dosages. Furthermore, models relating certain parameters to the dosage of chemicals have been derived. Corresponding parameters in the ASM2d and the secondary settler models, included in the Benchmark Simulation Model No 1 （BSM1）, have been modified to take the metal influence into consideration. Based on the effluent limits and penalty policy of China, an equivalent evaluation method was derived for the total cost assessment. A large number of 300-day steady-state and 14-day open-loop dynamic simulations were performed to demonstrate the difference in behavior between the original and the modified BSM1. The results show that 1） both in low and high mole concentrations, Fe^3＋ addition results in a higher phosphorus removal rate than Al^3＋; 2） the sludge settling velocity will increase due to the metal addition; 3） the respiration rate of the activated sludge is decreased more by the dosage of Al^3＋ than Fe^3＋; 4） the inhibition of Al^3＋ on the nitrification rate is stronger than that of Fe^3＋; 5） the total operating cost will reach the minimum point for smaller dosages of Fe^3＋, but always increase with Al^3＋ addition.

Characteristics of Nutrients Removal under Viscous Sludge Bulking Treating Synthetic Wastewater

Viscous sludge bulking is a rare phenomenon in activated sludge process.The performances of nutrients removal were investigated with normal sludge and viscous bulking sludge.The results showed that when COD loading and C/N ratio were around 0.13 mg COD/(mg MLSS·d)and 7.67,the effect of viscous sludge bulking on the maximum specific oxidation rates of NH_(4)^(+)⁃N was very little,while the maximum specific oxidation rates of NO_(2)^(-)⁃N decreased from 24.69 mg/(g·h)to 1.20 mg/(g·h).Compared with normal sludge,viscous bulking sludge had bigger particle size and more extracellular polymeric substances(EPS).The mass transfer resistance in sludge flocs might be the main cause of the difference in NO-2⁃N oxidation rates.Therefore,this study demonstrates that viscous sludge bulking is beneficial to enhance simultaneous nitrification and denitrification(SND),and excessive EPS will exhibit storage function during phosphorus removal process.

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.

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.

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.

Applying real-time control to enhance the performance of nitrogen removal in CAST system

A bench-scale reactor（72 L） red with domestic sewage, was operated more than 3 months with three operation modes： traditional mode, modified mode and real-time control mode, so as to evaluate effects of the operation mode on the system performance and to develop a feasible control strategy. Results obtained from fixed-time control study indicate that the variations of the pH and oxidation-reduction potential（ORP） profiles can represent dynamic characteristics of system and the cycle sequences can be controlled and optimized by the control points on the pH and ORP profiles. A control strategy was, therefore, developed and applied to real-time control mode. Compared with traditional mode, the total nitrogen（TN） removal can be increased by approximately 16% in modified mode and a mean TN removal of 92% was achieved in real-time control mode. Moreover, approximately 12.5% aeration energy was saved in real- time control mode. The result of this study shows that the performance of nitrogen removal was enhanced in modified operation mode. Moreover, the real-time control made it possible to optimize process operation and save aeration energy.

Simultaneous nitrogen and phosphorus removal under low dissolved oxygen conditions

A full-scale test was operated by using low dissolved oxygen activated sludge process to enhance biological nitrogen and phosphorus removal. When the influent concentrations of CODCr, TN and TP varied in a range of 352.9 mg/L-1338.2 mg/L, 34.4 mg/L-96.3 mg/L, and 2.21 mg/L-24.0 mg/L, the average removal efficiencies were 94.9%, 86.7% and 93.0%, respectively. During the test period of two months, effluent meas of CODCr,, BOD5, NH3-N, TN and TP were below 50 mg/L, 25 mg/L, 10 mg/L and 1.0 mg/L respectively. The low dissolved oxygen activated sludge process has a simple flow sheet, fewer facilities and high N and P removal efficiency. It is very convenient to retrofit the conventional activated sludge process with the above process.

Nitrogen and phosphorus removal under intermittent aeration conditions

A practice wastewater treatment plant was operated using intermittent aeration activated sludge process to enhance biological nitrogen and phosphorus removal. When the influent concentrations of COD Cr , BOD 5, TN, TP, NH 3\|N, TKN, and SS varied in a range of 207.5—1640 mg/L, 61.8—637 mg/L, 28.5—75.6 mg/L, 4.38—20.2 mg/L, 13.6—31.9 mg/L, 28.5—75.6 mg/L, and 111—1208 mg/L, the effluent means were less than 50 mg/L, 20 mg/L, 5 mg/L, 1.0 mg/L, 5 mg/L, 10 mg/L, and 20 mg/L, respectively. Based on a long time of operating results, this process is very suitable for nutrient biological removal for treating the municipal wastewater those water characteristics are similar as that of the Songjiang Municipal Waste Water Treatment Plant(SJMWTP).

A novel approach to estimate and control denitrification performance in activated sludge systems with respirogram technology

Respirogram technology has been widely applied for aerobic process, however, the response of respirogram to anoxic denitrification is still unclear. To reveal such response may help to design a new method for the evaluation of the performance of denitrification. The size distribution of flocs measured at different denitrification moments demonstrated a clear expansion of flocs triggered by denitrification, during which higher specific endogenous and quasi-endogenous respiration rates(SOUReand SOURq) were also observed. Furthermore,SOURqincreases exponentially with the specific denitrification rate(SDNR), suggesting that there should be a maximum SDNR in conventional activated sludge systems. Based on these findings, an index Rq/t, defined as the ratio of quasi-endogenous(OURq) to maximum respiration rate(OURt), is proposed to estimate the denitrification capacity that higher Rq/tindicates higher denitrification potential, which can be readily obtained without complex measurement or analysis, and it offers a novel and promising respirogram-based approach for denitrification estimation and control by taking measures to extend anoxic time to maintain its value at a high level within a certain range.

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.

污水生化处理工艺发展阶段化技术特征及未来趋势

百余年来,伴随人类社会和科技日新月异,污水生化处理技术也在实现快速发展与不断迭代,新工艺与新反应器、新功能微生物与新生化代谢途径不断被提出、发现和解析,并进一步推动了污水处理技术的进步与升级。回顾了过去百余年污水生化处理技术发展历程,就典型污水生化工艺,结合技术研发进展、技术成熟度及案例应用情况,给出了不同生化处理工艺代际划分与技术发展期“S曲线”,分析了近些年来新涌现出的一些革新性污水生化处理工艺的技术原理、技术特征,结合实际案例分析了工艺技术特征、技术优势与总体效能,从工艺强化、绿色低碳与集约高效等方面总结了未来污水处理发展的技术趋势,以期为“双碳背景”下排水系统提质增效工作、未来国内前瞻性污水厂工艺设计与运行提供参考和借鉴。