An observation on sludge granulation in an enhanced biological phosphorus removal process

A sequencing batch reactor (SBR) seeded with flocculated sludge and fed with synthetic wastewater was operated for an enhanced biological phosphorus removal (EBPR) process. Eight weeks after reactor startup, sludge granules were observed. The granules had a diameter of 0.5 to 3.0 mm and were brownish in color and spherical or ellipsoidal in shape. No significant change was observed in sludge granule size when operational pH was changed from 7 to 8. The 208-day continuous operation of the SBR showed that sludge granules were stably maintained with a sludge volume index (SVI) between 30 to 55 mL/g while securing a removal efficiency of 83% for carbon and 97% for phosphorus. Fluorescent in situ hybridization (FISH) confirmed the enrichment of polyphosphate accumulating organisms (PAOs) in the SBR. The observations of sludge granulation in this study encourage further studies in the development of granules-based EBPR process.     

Functionally relevant microorganisms to enhanced biological phosphorus removal performance at full-scale wastewater treatment plants in the United States

The abundance and relevance ofAccumulibacter phosphatis (presumed to be polyphosphate-accumulating organisms [PAOs]), Competibacter phosphatis (presumed to be glycogen-accumulating organisms [GAOs]), and tetrad-forming organisms (TFOs) to phosphorus removal performance at six full-scale enhanced biological phosphorus removal (EBPR) wastewater treatment plants were investigated. Coexistence of various levels of candidate PAOs and GAOs were found at these facilities. Accumulibacter were found to be 5 to 20% of the total bacterial population, and Competibacter were 0 to 20% of the total bacteria population. The TFO abundance varied from nondetectable to dominant. Anaerobic phosphorus (P) release to acetate uptake ratios (P(rel)/HAc(up)) obtained from bench tests were correlated positively with the abundance ratio of Accumulibacter/(Competibacter +TFOs) and negatively with the abundance of (Competibacter +TFOs) for all plants except one, suggesting the relevance of these candidate organisms to EBPR processes. However, effluent phosphorus concentration, amount of phosphorus removed, and process stability in an EBPR system were not directly related to high PAO abundance or mutually exclusive with a high GAO fraction. The plant that had the lowest average effluent phosphorus and highest stability rating had the lowest P(rel)/HAc(up) and the most TFOs. Evaluation of full-scale EBPR performance data indicated that low effluent phosphorus concentration and high process stability are positively correlated with the influent readily biodegradable chemical oxygen demand-to-phosphorus ratio. A system-level carbon-distribution-based conceptual model is proposed for capturing the dynamic competition between PAOs and GAOs and their effect on an EBPR process, and the results from this study seem to support the model hypothesis.     

A practical method for quantification of phosphorus- and glycogen-accumulating organism populations in activated sludge systems.

Enhanced biological phosphorus removal (EBPR) from wastewater relies on the enrichment of activated sludge with phosphorus-accumulating organisms (PAOs). The presence and proliferation of glycogen-accumulating organisms (GAOs), which compete for substrate with PAOs, may be detrimental for EBPR systems, leading to deterioration and, in extreme cases, failure of the process. Therefore, from both process evaluation and modeling perspectives, the estimation of PAO and GAO populations in activated sludge systems is a relevant issue. A simple method for the quantification of PAO and GAO population fractions in activated sludge systems is presented in this paper. To develop such a method, the activity observed in anaerobic batch tests executed with different PAO/GAO ratios, by mixing highly enriched PAO and GAO cultures, was studied. Strong correlations between PAO/GAO population ratios and biomass activity were observed (R2 > 0.97). This served as a basis for the proposal of a simple and practical method to quantify the PAO and GAO populations in activated sludge systems, based on commonly measured and reliable analytical parameters (i.e., mixed liquor suspended solids, acetate, and orthophosphate) without requiring molecular techniques. This method relies on the estimation of the total active biomass population under anaerobic conditions (PAO plus GAO populations), by measuring the maximum acetate uptake rate in the presence of excess acetate. Later, the PAO and GAO populations present in the activated sludge system can be estimated, by taking into account the PAO/GAO ratio calculated on the basis of the anaerobic phosphorus release-to-acetate consumed ratio. The proposed method was evaluated using activated sludge from municipal wastewater treatment plants. The results from the quantification performed following the proposed method were compared with direct population estimations carried out with fluorescence in situ hybridization analysis (determining Candidatus Accumulibacter Phosphatis as PAO and Candidatus Competibacter Phosphatis as GAO). The method showed to be potentially suitable to estimate the PAO and GAO populations regarding the total PAO-GAO biomass. It could be used, not only to evaluate the performance of EBPR systems, but also in the calibration of potential activated sludge mathematical models, regarding the PAO-GAO coexistence.     

基于不同基质的强化生物除磷系统中生化反应机理研究进展

阐述了3种不同基质(乙酸盐、丙酸盐和葡萄糖)在强化生物除磷系统中的生化模型反应机理.重点介绍了聚磷菌(PAOs)、聚糖菌(GAOs)和产乳酸菌在厌氧/好氧条件下对能量及还原力(NADH2)的利用方式;聚-β-羟基链烷酸盐(PHA)的合成方式及存在形式,糖原、乳酸(L型)的代谢途径等.虽然控制基质条件可以实现稳定的强化生物除磷效果,但目前的生化模型并不能完全解释所有的代谢过程,今后要在分离纯种的PAOs及相关生化代谢过程中酶的活动等方面进行深入研究.     

The selective role of nitrite in the PAO/GAO competition

Proliferation of Glycogen Accumulating Organisms (GAOs) accounts as one of the major bottlenecks in biological phosphorus removal systems. GAO outcompeting polyphosphate accumulating organisms (PAOs) results in lower P-removal. Thus, finding optimal conditions that favour PAO in front of GAO is a current focus of research. This work shows how nitrite can provide a novel strategy for PAO enrichment. A propionate-fed GAO-enriched biomass (70% Defluviicoccus I, 18% Defluviicoccus II and 10% PAO) was subjected more than 50 d under anaerobic-anoxic conditions with nitrite as electron acceptor. These operational conditions led to a PAO-enriched sludge (85%) where GAO were washed out of the system (<10%), demonstrating the validity of the new approach for PAO enrichment. In addition, the presented suppression of Defluviicocus GAO with nitrite represents an add-on benefit to the nitrite-based systems since the proliferation of non-desirable GAO can be easily ruled out and added to the other benefits (i.e. lower aeration and COD requirements).     

不同电子受体对聚磷菌吸磷性能的影响研究

采用人工配水,在厌氧/好氧交替运行的序批式活性污泥反应器(SBR)中,富集了全菌数量80%以上的聚磷菌(Candi-datus Accumulibacter Phosphates)。以此为基础,研究了O2及不同浓度NO3--N、NO2--N对聚磷菌吸磷的影响。结果表明,在一定的条件下,聚磷菌可以NO3--N和NO2--N为电子受体进行缺氧吸磷;NO3--N浓度对聚磷菌的吸磷速率影响很小;聚磷菌可以低质量浓度NO2--N(≤40mg/L)为电子受体,但不能以高质量浓度NO2--N(≥80mg/L)为电子受体,而且高浓度NO2--N对聚磷菌吸磷产生抑制甚至对细菌本身存在毒害;NO2--N为电子受体时,其抑制浓度和污泥本身以及外界条件都存在很大的关系,各个研究结论不尽相同,其影响过程有待进一步的探讨。     

The mechanism of enhanced biological phosphorus removal washout and temperature relationships.

In this study, the combined effects of temperature and solids retention time (SRT) on enhanced biological phosphorus removal (EBPR) performance and the mechanism of EBPR washout were investigated. Two pilot-scale University of Cape Town (South Africa) systems fed with synthetic wastewater were operated at 5 and 10 degrees C. The results showed that the phosphorus removal performance was optimum at total SRT ranges of 16 to 24 days and 12 to 17 days for 5 and 10 degrees C, respectively, and steady-state phosphorus removal was greater at the lower temperature. Higher SRT values of up to 32 days at 5 degrees C and 25 days at 10 degrees C slightly reduced EBPR performance as a result of increased extent of endogenous respiration, which consumed internally stored glycogen, leaving less reducing power for poly-hydroxy alkanoate (PHA) formation in anaerobic stages. The phosphorus-accumulating organism (PAO) washout SRTs of the systems were determined as 3.5 days at 5 degrees C and 1.8 days at 10 degrees C, considerably less than the washout SRTs of nitrifiers. Polyphosphorus, the main energy reserve of the EBPR bacterial consortium, was not completely depleted, even at washout points. The inability of EBPR biomass to use glycogen to generate reducing power for PHA formation was the major reason for washout. The results not only suggest that glycogen mechanism is the most rate-limiting step in EBPR systems, but also that it is an integral part of EBPR biochemistry, as proposed originally by Mino et al. (1987), and later others (Pereira et al., 1996, Erdal et al., 2002; Erdal, Z. K., 2002). The aerobic washout SRT values (2.1 and 1.2 days for 5 and 10 degrees C, respectively) of this study did not fit the linear line for PAO washout developed by Mamais and Jenkins (1992). Perhaps this was because the feeds used during this study were chemical-oxygen-demand-limited (acetate-based synthetic feed), whereas the feeds used for their study were phosphorus-limited (external acetate added to domestic wastewater), resulting in different ratios of PAOs and nonPAOs in the biomass.     

活性污泥体系中聚糖菌的富集与鉴定

活性污泥体系中,聚糖菌(GAOs)在厌氧环境下与聚磷菌(PAOs)形成对底物的竞争关系,对聚糖菌的研究对于优化生物除磷工艺有重要意义。以葡萄糖为惟一碳源,在磷限制条件下,利用特殊运行方式对活性污泥进行驯化培养出了稳定的聚糖菌颗粒污泥,厌氧阶段磷释放量与有机物吸收量浓度(mg/L)比从7.4%下降为0.25%。从培养好的活性污泥反应器中分离获得2株聚糖菌,经菌落PCR和16S rRNA序列分析确定了所得聚糖菌菌株G1和菌株G2分别是枯草芽孢杆菌(Bacillus subtilis)和解鸟氨酸克雷伯氏菌(Klebsiella ornithinolytica)。     

Modeling the aerobic metabolism of polyphosphate-accumulating organisms enriched with propionate as a carbon source.

In enhanced biological phosphorus removal (EBPR) systems, polyphosphate-accumulating organisms (PAOs) are primarily responsible for removing phosphate from wastewater. Propionate is an abundant carbon substrate in many EBPR plants and has been suggested to provide PAOs an advantage over their carbon competitors--the glycogen-accumulating organisms (GAOs). The aerobic metabolism of PAOs enriched with a propionate carbon source is studied in this paper. A metabolic model is proposed and experimentally validated to characterize the aerobic biochemical transformations by PAOs. The model predicts very well the experimental data obtained from the enriched PAO culture through solid-, liquid-, and gas-phase analyses. This model may be combined with previously formulated metabolic models to better describe the biochemical activity of PAOs with acetate and propionate as the primary carbon sources. Furthermore, it can also facilitate the study of the effect of different carbon sources on PAO-GAO competition.     

Effect of initial pH control on biological phosphorus removal induced by the aerobic/extended-idle regime

Recently, it has been reported that biological phosphorus removal (BPR) can be induced by an aerobic/extended-idle (AEI) regime. This study further investigated the effect of initial pH ranging from 6.6 to 8.2 on BPR in the AEI process, and compared the BPR performance between the AEI and the anaerobic/oxic (A/O) regimes under their optimal initial pH value. Experimental results firstly showed that phosphorus removal linearly increased with initial pH increasing from 6.6 to 7.8, but slightly decreased when initial pH increased from 7.8 to 8.2. The optimal initial pH should be controlled at 7.8, and the phosphorus removal at initial pH 7.8 was approximately 1.7-time than that at initial pH 6.6. The mechanism studies showed that the biomass cultured at initial pH 7.8 contained more polyphosphate accumulating organisms (PAOs), lower glycogen accumulating organisms (GAOs), and had higher activities of exopolyphosphatase and polyphosphate kinase than that cultured at initial pH 6.6. Cyclic studies revealed that initial pH control affected the transformations of intracellular polyhydroxyalkanoates and glycogen, which might thereby influence microbial competition between PAOs and GAOs. Then, BPR performance between the AEI and the A/O regimes by adjusting initial pH at 7.8 was also compared. The results showed the AEI regime could drive a better BPR than the generally accepted A/O regime (98% vs 88%). Finally, controlling initial pH at 7.8 to promote BPR in the AEI process was confirmed for a municipal wastewater.     

Molecular characterization of denitrifying bacteria isolated from the anoxic reactor of a modified DEPHANOX plant performing enhanced biological phosphorus removal

Enhanced Biological Phosphorus Removal (EBPR) under anoxic conditions was achieved using a Biological Nutrient Removal (BNR) system based on a modification of the DEPHANOX configuration. Double-probe Fluorescence in Situ Hybridization (FISH) revealed that Polyphosphate Accumulating Organisms (PAOs) comprised 12.3 +/- 3.2% of the total bacterial population in the modified DEPHANOX plant. The growing bacterial population on blood agar and Casitone Glycerol Yeast Autolysate agar (CGYA) medium was 16.7 +/- 0.9 x 10(5) and 3.0 +/- 0.6 x 10(5) colony forming units (cfu) mL(-1) activated sludge, respectively. A total of 121 bacterial isolates were characterized according to their denitrification ability, with 26 bacterial strains being capable of reducing nitrate to gas. All denitrifying isolates were placed within the alpha-, beta-, and gamma-subdivisions of Proteobacteria and the family Flavobacteriaceae. Furthermore, a novel denitrifying bacterium within the genus Pseudomonas was identified. This is the first report on the isolation and molecular characterization of denitrifying bacteria from EBPR sludge using a DEPHANOX-type plant.     

Understanding the granulation process of activated sludge in a biological phosphorus removal sequencing batch reactor

The granulation of activated sludge was investigated using two parallel sequencing batch reactors (SBRs) operated in biological nitrogen and phosphorus removal conditions though the reactor configuration and operating parameters did not favor the granulation. Granules were not observed when the SBR was operated in biological nitrogen removal period for 30 d. However, aerobic granules were formed naturally without the increase of aeration intensity when enhanced biological phosphorus removal (EBPR) was achieved. It can be detected that plenty of positive charged particles were formed with the release of phosphorus during the anaerobic period of EBPR. The size of the particles was about 5-20 μm and their highest positive ζ potential was about 73 mV. These positive charged particles can stimulate the granulation. Based on the experimental results, a hypothesis was proposed to interpret the granulation process of activated sludge in the EBPR process in SBR. Dense and compact subgranules were formed stimulated by the positive charged particles. The subgranules grew gradually by collision, adhesion and attached growth of bacteria. Finally, the extrusion and shear of hydrodynamic shear force would help the maturation of granules. Aerobic granular SBR showed excellent biological phosphorus removal ability. The average phosphorus removal efficiency was over 95% and the phosphorus in the effluent was below 0.50 mg L⁻¹ during the operation.     

原位臭氧氧化污泥减量工艺的运行效能

采用ASBR/SBR原位臭氧污泥减量工艺,重点研究了原位臭氧氧化对SBR段污泥产率和出水水质的影响。两个相同的ASBR/SBR组合工艺同时运行,每隔3个周期向臭氧投加组SBR的曝气阶段原位间歇投加臭氧,臭氧投加量为0.027 g O3/g MLSS,连续运行40 d;对照组不投加臭氧作为对比。结果表明,原位臭氧氧化实现污泥减量约43.9%,臭氧投加组SBR段平均污泥产率系数为0.1447 g SS/g SCOD,而对照组为0.2580 g SS/g SCOD,投加组没有惰性污泥的累积,并且污泥沉淀性能得到改善。原位臭氧氧化对出水水质影响不大,投加组与对照组相比,臭氧投加3周期后的出水COD、NH4+-N、TN和TP平均值分别为47.8、0.76、14.1和6.4 mg/L,去除率分别下降了4%、2%、3%和7.7%,其中COD、NH4+-N和TN均能达到《城镇污水处理厂污染物排放标准》(GB18918-2002)一级A标准。     

反硝化聚磷菌群的培养驯化

以污水处理厂二沉池的活性污泥为种泥,采用SBR反应器初步完成了反硝化聚磷菌(DPB)的培养与驯化.在第1阶段的30 d里,污泥进行了厌氧-好氧驯化,聚磷菌好氧吸磷最终可基本稳定在85%左右,然后转变驯化条件进行第2阶段的厌氧-缺氧驯化,60 d后磷的去除率稳定在70%左右.通过实验得出,硝酸盐的消耗量与磷的吸收量基本呈线性关系,认为系统基本完成了污泥的驯化.     

Behaviors of intercellular materials and nutrients in biological nutrient removal process supplied with domestic wastewater and food waste.

A four-stage biological nutrient removal (BNR) process was operated to investigate the effect of anaerobically fermented leachate of food waste (AFLFW) as an external carbon source on nutrient removal from domestic wastewater having a low carbon-to-nitrogen ratio. The BNR system that was supplemented with AFLFW showed a good performance at a sludge retention time (SRT) of 30 days, despite low temperature. With this wastewater, average removal efficiencies of soluble chemical oxygen demand (COD), total nitrogen (T-N), and total phosphorus (T-P) were 88 to 93%, 70 to 74%, and 63 to 68%, respectively. In this study, several kinds of poly-hydroxyalkanoates (PHAs) were observed in cells. These included 24% poly-3-hydroxybutyrate (PHB), 41% poly-3-hydroxyvalerate (PHV), 18% poly-3-hydroxyhexanoate (PHH), 10% poly-3-hydroxyoctanoate (PHO), 5% poly-3-hydroxydecanoate (PHD). and 2% poly-3-hydroxydodecanoate (PHDD), indicating that microorganisms could store various PHAs through the different metabolic pathways. However, breakdown of the enhanced biological phosphorus removal (EBPR) mechanism was observed when SRT increased from 30 to 50 days for the enhancement of nitrification. To study the effect of SRT on EBPR, a sequencing batch reactor (SBR) system that was supplied with glucose was operated at various SRTs of 5, 10, and 15 days. Nitrification and denitrification efficiencies increased as SRT increased. However, the content of intracellular materials such as PHAs, glycogen. and poly-P in cells decreased. From these results, it was concluded that SRT should be carefully controlled to increase nitrification activity and to maintain biological phosphorus removal activity in the BNR process.     

Influence of chemical oxygen demand/total Kjeldahl nitrogen ratio and sludge age on nitrification of nitrogenous wastewater.

Four laboratory-scale biological nitrification units (influent total Kjeldahl nitrogen [TKN] = 1002 to 1062 mg/L) were operated at chemical oxygen demand (COD)/TKN ratios of approximately 0.5, 1.0,15, and 2.0 and at three different sludge ages of 30, 20, and 10 days to study the influence of COD/TKN, sludge age, COD loading, and TKN loading on nitrification and nitrifiers. Percent nitrification was found to increase with decreases in COD/TKN and increases in sludge age. The average nitrifier concentration increased from 460 mg/L at a COD/TKN of 2.22 and a sludge age of 10 days to 706 mg/L at a COD/TKN of 0.676 and a sludge age of 30 days. The nitrifier fraction was found to be higher at a lower COD/TKN and lower at a higher COD/TKN. The nitrifier fraction increased with the decrease in sludge age and COD loadings and the increase in TKN loadings. The effect of sludge age on the nitrifier fraction was amplified at a COD/ TKN of approximately 0.5 rather than at approximately 2.0. The nitrification rate (kilograms TKN oxidized per kilograms nitrifiers per day) was shown to be dependent on COD/TKN and sludge age. The activity performed by Nitrobacter was affected at all COD/TKN ratios studied as well as at a sludge age of 10 days. This was manifested by the accumulation of high levels of nitrite-nitrogen in the nitrified effluent. The presence of heterotrophs did not affect nitrification rates and the growth of nitrifiers, which were found to be beneficial. High sludge age and COD loadings resulted in a higher sludge volume index of more than 200 mL/g mixed liquor suspended solids. Microscopic examination showed filamentous structure of sludge under these conditions. It is concluded from the investigations that a sludge age of 30 days and a COD/TKN of approximately 1.0 are optimal to yield maximum nitrification and nitrifier growth rates for treating high-strength nitrogenous wastewater.     

Effect of dissolved oxygen on biological nutrient removal by denitrifying phosphorus-accumulating organisms in a continuous-flow system

A laboratory-scale continuous-flow system with an anaerobic/anoxic/aerobic configuration was set up to study the effect of oxygen in the internal recycle stream; of particular interest was its performance of denitrifying phosphorus-accumulating organisms (DPAOs). It was found that, by using a degas device, the dissolved oxygen in the nitrate recycle stream was effectively decreased from 0.1 +/- 0.02 to 0.01 +/- 0.01 mg/L. This provided a favorable condition for DPAOs to grow under an anoxic condition and thus be sustained successfully in the system. When the degas device was removed from the system, the dissolved oxygen concentration in the anoxic reactor increased to 0.1 +/- 0.02 mg/L. The proliferation of the denitrifying glycogen-accumulating organisms (DGAOs) population and deterioration of DPAOs performance was observed. The increased population of DGAO/GAOs, which competed for the carbon source with DPAO/ PAOs, resulted in a poor performance of biological phosphorus removal.     

畜禽养殖废水生物处理剩余污泥臭氧减量过程中的重金属释放

采用臭氧氧化污泥减量法对畜禽养殖废水SBR中的剩余污泥进行处理。当臭氧反应时间控制在30min时,污泥的溶解比例在30％左右（以MLSS计）。上清液中一定量的SCOD溶出可为生物处理单元提供充足的碳源,同时在上清液中,TN、TP及水相重金属浓度增加有限,臭氧氧化后的污泥液回流污水处理系统后造成的N、P处理负荷较小,重金属对污泥微生物的活性抑制风险较低。若继续延长臭氧的反应时间,上清液SCOD、TN、TP以及重金属cu、Pb的释放速率明显增加,同时上清液的C／N降低,臭氧化后的污泥液回流反而不利于生物单元的脱氮处理。综合考虑TN、TP及水相重金属浓度增加的危害性,臭氧反应时间应控制在30min,臭氧实际投加量应为123．1mg O3／gSS。     

Full-scale use of glycogen-accumulating organisms for excess biological carbon removal

The purpose of this study has been to verify the efficient full-scale applicability of glycogen-accumulating organisms (GAOs) for excess biological carbon removal, that is, for removing more carbon substrate than the amount of available nutrients would allow in the conventional activated sludge process of microbial growth. This aims to cost-effectively overcome the problem of viscous bulking occurring in a fully aerated system, with nutrient deficiency. Analytical data measured at the wastewater treatment plant of the Balatonboglár (BB) winery in Balatonboglár, Hungary, containing consecutive unaerated and aerated activated sludge basins, reflected a high performance with efficient carbon removal and good sludge settling, without dosing any external nutrient source to the severely nitrogen- and phosphorous-deficient influent. Supplementary laboratory-scale batch experiments and microbiological tests verified the abundance of GAOs in the activated sludge system and elucidated their role in efficient excess biological carbon removal.     

利用亚硝酸盐为电子受体反硝化聚磷菌的筛选与富集

依据DPB原理,利用SBR动态反应器和静态释/聚磷装置.以A2/O工艺厌氧段污泥为种泥,研究以亚硝酸盐为电子受体反硝化聚磷菌的筛选与富集,同时对选择、富集污泥的反硝化聚磷性能进行了考察.结果表明:利用亚硝酸盐为电子受体的反硝化聚磷菌存在于A2/O厌氧段污泥中,通过厌氧/好氧和厌氧/缺氧方式运行后,聚磷菌总数由1400个/mL增加到32 000个/mL,其中反硝化聚磷菌占聚磷菌总数的比例也由14.5%提高到81%,磷酸盐和亚硝酸盐去除率分别由最初的8.65%和7.55%上升到91%和95.62%;筛选与富集利用亚硝酸盐为电子受体的反硝化聚磷菌时,缺氧段进水COD的浓度须控制在10 mg/L以下;当体系处于稳定状态,且亚硝酸盐氮浓度高达30 mg/L时,并未对反硝化聚磷菌的生存产生抑制和体系运行产生干扰,此时磷酸盐出水低至1.06 mg/L.