Long-term impact of anaerobic reaction time on the performance and granular characteristics of granular denitrifying biological phosphorus removal systems

Removal of nitrogen and phosphorus (P) from wastewater is successfully and widely practiced in systems employing both granular sludge technology and enhanced biological P removal (EBPR) processes; however, the key parameter, anaerobic reaction time (AnRT), has not been thoroughly investigated. Successful EBPR is highly dependent on an appropriate AnRT, which induces carbon and polyphosphate metabolism by phosphorus accumulating organisms (PAOs). Therefore, the long-term impact of AnRT on denitrifying P removal performance and granular characteristics was investigated in three identical granular sludge sequencing batch reactors with AnRTs of 90 (R1), 120 (R2) and 150 min (R3). The microbial community structures and anaerobic stoichiometric parameters related to various AnRTs were monitored over time. Free nitrite acid (FNA) accumulation (e.g., 0.0008–0.0016 mg HNO₂–N/L) occurred frequently owing to incomplete denitrification in the adaptation period, especially in R3, which influenced the anaerobic/anoxic intracellular intermediate metabolites and activities of intracellular enzymes negatively, resulting in lower levels of poly-P and reduced activity of polyphosphate kinase. As a result, the Accumulibacter-PAOs population decreased from 51 ± 2.5% to 43 ± 2.1% when AnRT was extended from 90 to 150 min, leading to decreased denitrifying P removal performance. Additionally, frequent exposure of microorganisms to the FNA accumulation and anaerobic endogenous conditions in excess AnRT cases (e.g., 150 min) stimulated increased extracellular polymeric substances (EPS) production by microorganisms, resulting in enhanced granular formation and larger granules (size of 0.6–1.2 mm), but decreasing anaerobic PHA synthesis and glycogen hydrolysis. Phosphorus removal capacity was mediated to some extent by EPS adsorption in granular sludge systems that possessed more EPS, longer AnRT and relatively higher GAOs.     

Advancing post-anoxic denitrification for biological nutrient removal

The objective of this research was to advance a fundamental understanding of a unique post-anoxic denitrification process for achieving biological nutrient removal (BNR), with an emphasis on elucidating the impacts of surface oxygen transfer (SOT), variable process loadings, and bioreactor operational conditions on nitrogen and phosphorus removal. Two sequencing batch reactors (SBRs) were operated in an anaerobic/aerobic/anoxic mode for over 250 days and fed real municipal wastewater. One SBR was operated with a headspace open to the atmosphere, while the other had a covered liquid surface to prevent surface oxygen transfer. Process performance was assessed for mixed volatile fatty acid (VFA) and acetate-dominated substrate, as well as daily/seasonal variance in influent phosphorus and ammonia loadings. Results demonstrated that post-anoxic BNR can achieve near-complete (>99%) inorganic nitrogen and phosphorus removal, with soluble effluent concentrations less than 1.0 mgN L⁻¹ and 0.14 mgP L⁻¹. Observed specific denitrification rates were in excess of typical endogenous values and exhibited a linear dependence on the glycogen concentration in the biomass. Preventing SOT improved nitrogen removal but had little impact on phosphorus removal under normal loading conditions. However, during periods of low influent ammonia, the covered reactor maintained phosphorus removal performance and showed a greater relative abundance of polyphosphate accumulating organisms (PAOs) as evidenced by quantitative real-time PCR (qPCR). While GAOs were detected in both reactors under all operational conditions, BNR performance was not adversely impacted. Finally, secondary phosphorus release during the post-anoxic period was minimal and only occurred if nitrate/nitrite were depleted post-anoxically.     

新型双泥生物反硝化除磷脱氮工艺

在对生物脱氮与除磷机理进行深入研究后发现，生物脱氮与除磷是两个相对独立而又相互交叉的生理过程，其交叉点是部分聚磷菌在缺氧状态下的反硝化吸磷脱氮。在此基础上提出的新型双泥生物反硝化除磷脱氮工艺（由两个不同功能的SBR反应器组成）成功地解决了硝化菌与聚磷菌的泥龄之争。反硝化与聚磷菌厌氧释磷的矛盾等难题，该工艺运行稳定且处理效果良好，特别适合于处理BOD5/TP值低的污水。     

Development of a 2-sludge, 3-stage system for nitrogen and phosphorous removal from nutrient-rich wastewater using granular sludge and biofilms

A novel 2-sludge 3-stage process using a combination of granular sludge and biofilm was developed to achieve biological removal of nitrogen and phosphorus from nutrient-rich wastewater. The system consists of a granular sequencing batch reactor (SBR) working under alternating anaerobic/anoxic conditions supplemented with a short aerobic phase and an aerobic biofilm SBR. The wastewater is first fed to the granular SBR reactor, where easily biodegradable carbon sources are taken up primarily by polyphosphate accumulating organisms (PAOs). The supernatant resulting from quick settling of the granular sludge is then fed to the biofilm SBR for nitrification, which produces oxidized nitrogen that is returned to the granular reactor for simultaneous denitrification and phosphorus removal. While maximizing the utilization of organic substrates and reducing operational costs, as do other 2-sludge processes previously reported in literature, the proposed system solves the bottleneck problem of traditional 2-sludge systems, namely high effluent ammonia concentration, due to its high-volume exchange ratios. An ammonia oxidation rate of 32 mg N/Lh was achieved in the biofilm SBR, which produced nitrite as the final product. This nitrite stream was found to cause major inhibition on the anoxic P uptake and also to result in the accumulation of N(2)O. These problems were solved by feeding the nitrite-containing stream continuously to the granular reactor in the anoxic phase. With a nitrogen and phosphorus removal efficiency of 81% and 94%, respectively, the system produces an effluent that is suitable for land irrigation from a wastewater stream containing 270 mg N/L of total nitrogen and 40 mg P/L of total phosphorus.     

亚硝酸盐对聚磷菌厌氧代谢的影响

以2种强化生物除磷(EBPR)系统中的活性污泥为研究对象,考察亚硝酸盐对聚磷菌厌氧代谢的影响,结果表明：不同EBPR系统中的聚磷菌对于亚硝酸盐的耐受能力不同。人工配水富集聚磷菌的活性污泥,当亚硝态氮浓度超过10 mg/L时,聚磷菌吸收VFA受到抑制, PHA的合成减少,磷酸盐的释放增加;处理生活污水的SBR短程脱氮除磷活性污泥,亚硝酸盐的浓度高达30 mg/L时,未对聚磷菌的厌氧代谢造成抑制,但引起异养反硝化菌与聚磷菌竞争VFA,导致PHA合成量和释磷量的减少。富集聚磷菌的活性污泥投加亚硝酸盐后P/VFA     

污水强化除磷工艺的现状与未来

对强化生物除磷工艺的基本概念、外加碳源的作用、脱氮除磷的关系、环境影响参数进行了阐述,并介绍了聚磷菌菌群研究的最新进展。最后探讨了如何通过生产鸟粪石(MAP)实现磷元素回收,并围绕污水除磷和回收磷这一课题,提出了研究方向。     

Advances in enhanced biological phosphorus removal: from micro to macro scale

The enhanced biological phosphorus removal (EBPR) process has been implemented in many wastewater treatment plants worldwide. While the EBPR process is indeed capable of efficient phosphorus (P) removal performance, disturbances and prolonged periods of insufficient P removal have been observed at full-scale plants on numerous occasions under conditions that are seemingly favourable for EBPR. Recent studies in this field have utilised a wide range of approaches to address this problem, from studying the microorganisms that are primarily responsible for or detrimental to this process, to determining their biochemical pathways and developing mathematical models that facilitate better prediction of process performance. The overall goal of each of these studies is to obtain a more detailed insight into how the EBPR process works, where the best way of achieving this objective is through linking together the information obtained using these different approaches. This review paper critically assesses the recent advances that have been achieved in this field, particularly relating to the areas of EBPR microbiology, biochemistry, process operation and process modelling. Potential areas for future research are also proposed. Although previous research in this field has undoubtedly improved our level of understanding, it is clear that much remains to be learned about the process, as many unanswered questions still remain. One of the challenges appears to be the integration of the existing and growing scientific knowledge base with the observations and applications in practice, which this paper hopes to partially achieve.     

Incorporating microbial ecology into the metabolic modelling of polyphosphate accumulating organisms and glycogen accumulating organisms

In the enhanced biological phosphorus removal (EBPR) process, the competition between polyphosphate accumulating organisms (PAO) and glycogen accumulating organisms (GAO) has been studied intensively in recent years by both microbiologists and engineers, due to its important effects on phosphorus removal performance and efficiency. This study addresses the impact of microbial ecology on assessing the PAO-GAO competition through metabolic modelling, focussing on reviewing recent developments, discussion of how the results from molecular studies can impact the way we model the process, and offering perspectives for future research opportunities based on unanswered questions concerning PAO and GAO metabolism. Indeed, numerous findings that are seemingly contradictory could in fact be explained by the metabolic behaviour of different sub-groups of PAOs and/or GAOs exposed to different environmental and operational conditions. Some examples include the glycolysis pathway (i.e. Embden-Meyerhof-Parnas (EMP) vs. Entner-Doudoroff (ED)), denitrification capacity, anaerobic tricarboxylic acid (TCA) cycle activity and PAOs’ ability to adjust their metabolism to e.g. a GAO-like metabolism. Metabolic modelling may further yield far-reaching influences on practical applications as well, and serves as a bridge between molecular/biochemical research studies and the optimisation of wastewater treatment plant operation.     

生物/化学组合工艺处理高盐榨菜废水的除磷效能

针对高盐度废水生物除磷的难点问题,采用生物/化学组合工艺处理高盐度、高磷、高氮的榨菜腌制废水,考察了运行工况、挂膜密度、排泥周期、药剂种类和投加量等对除磷效能的影响.试验结果表明:采用厌氧/生物除磷/生物脱氮/化学除磷组合工艺除磷高效、可行,当一级SBBR生物除磷单元的挂膜密度为60%、排泥周期为2 d、运行工况为进水(O.2 h)-厌氧(3 h)-好氧(6 h)-沉淀及排水(0.2 h),化学除磷单元按物质的量之比为9:1投加硫酸铝时,在进水COD及(PO3-4)-P分别为10 000 mg/L和38.5 mg/L的条件下,出水COD和(PO3-4)-P分别为90和0.1mg/L,去除率均达到了99%以上.生物除磷、生物脱氮、化学除磷单元的除磷分担率分别为56.6%、20.8%和22%.     

Alumina nanoparticles-induced effects on wastewater nitrogen and phosphorus removal after short-term and long-term exposure

Alumina nanoparticles (Al₂O₃ NPs) have been widely used in many fields, which causes a growing concern about their potential health and environmental risks. However, their possible impacts on wastewater nitrogen and phosphorus removal have not yet been reported. In this study, both short-term and long-term effects of Al₂O₃ NPs on wastewater nutrient removal were investigated. Scanning electron microscope (SEM) analysis showed that most of Al₂O₃ NPs were adsorbed onto activated sludge, but these NPs had no adverse effects on the surface integrity and viability of activated sludge. It was found that short-term exposure to 1 and 50 mg/L Al₂O₃ NPs induced marginal influences on wastewater nitrification, denitrification and phosphorus removal. Nevertheless, the prolonged exposure to 50 mg/L Al₂O₃ NPs was observed to decrease the total nitrogen (TN) removal efficiency from 80.4% to 62.5% due to the suppressed denitrification process, although biological phosphorus removal and the transformations of intracellular polyhydroxyalkanoates and glycogen were not affected. Quantitative PCR assays indicated that compared with the control, 50 mg/L Al₂O₃ NPs decreased the abundance of denitrifying bacteria in activated sludge. Further enzyme activity tests showed that the activities of key denitrifying enzymes (nitrate reductase and nitrite reductase) were inhibited, which might be responsible for the negative effects of 50 mg/L Al₂O₃ NPs on wastewater nitrogen removal after long-term exposure.     

Phosphorus recovery by struvite crystallization in WWTPs: Influence of the sludge treatment line operation

Phosphorus recovery by struvite (MgNH₄PO₄·6H₂O) crystallization is one of the most widely recommended technologies for treating sludge digester liquors especially in wastewater treatments plants (WWTP) with enhanced biological phosphorus removal (EBPR). In this paper, phosphorus recovery by struvite crystallization is assessed using the rejected liquors resulting from four different operational strategies of the sludge treatment line. Phosphorus precipitation and recovery efficiencies of between 80-90% and 70-85%, respectively, were achieved in the four experiments. The precipitates formed were mainly struvite, followed by amorphous calcium phosphate and, in some experiments, by calcite. The highest global phosphorus recovery taking into account both the sludge line and the crystallizer was achieved when mixed thickening and high elutriation were carried out (8.4 gP/kg treated sludge). However, low struvite content was obtained in the crystallizer with this operation scheme due to the high calcium content in the elutriation stream. Therefore, if the final purpose is to obtain struvite, the reduction of the elutriation flowrate is widely recommended in the case of high water hardness.     

Nitrogen and phosphorus removal from municipal wastewater effluent using microalgal biofilms

Microalgal biofilms have so far received little attention as post-treatment for municipal wastewater treatment plants, with the result that the removal capacity of microalgal biofilms in post-treatment systems is unknown. This study investigates the capacity of microalgal biofilms as a post-treatment step for the effluent of municipal wastewater treatment plants. Microalgal biofilms were grown in flow cells with different nutrient loads under continuous lighting of 230 μmol/m²/s (PAR photons, 400-700 nm). It was found that the maximum uptake capacity of the microalgal biofilm was reached at loading rates of 1.0 g/m²/day nitrogen and 0.13 g/m²/day phosphorus. These maximum uptake capacities were the highest loads at which the target effluent values of 2.2 mg/L nitrogen and 0.15 mg/L phosphorus were still achieved. Microalgal biomass analysis revealed an increasing nitrogen and phosphorus content with increasing loading rates until the maximum uptake capacities. The internal nitrogen to phosphorus ratio decreased from 23:1 to 11:1 when increasing the loading rate. This combination of findings demonstrates that microalgal biofilms can be used for removing both nitrogen and phosphorus from municipal wastewater effluent.     

Performance of IFAS wastewater treatment processes for biological phosphorus removal

Integrated fixed film activated sludge (IFAS) is a promising process for the enhancement of nitrification and denitrification in conventional activated sludge systems that need to be upgraded for biological nutrient removal (BNR), particularly when they have space limitations or need modifications that will require large monetary expenses. Several studies have reported successful implementations of IFAS at temperate zone wastewater treatment facilities, typically by placement of fixed film media into aerobic zones. However, nearly all of the implementations have not included enhanced biological phosphorus removal (EBPR) in the upgraded systems. This is possibly because the treatment plants have been operated at low mixed liquor mean cell residence times (MCRTs), and EBPR would wash out of the systems at the low temperatures encountered, making it difficult to maintain EBPR. The primary objective of this study was to investigate the incorporation of EBPR into IFAS systems, and study the interactions between the fixed biomass and the mixed liquor suspended solids with respect to substrate competition and nutrient removal efficiencies. Three pilot-scale UCT/VIP configuration systems were used, one as a control and the other two with Bioweb media integrated into some of the anoxic and aerobic reactors. The systems were operated at different MCRTs, and influent COD/TP ratios, and with split influent flows. The experimental results confirmed that EBPR could be incorporated successfully into IFAS systems, but the redistribution of biomass resulting from the integration of fixed film media, and the competition of organic substrate between EBPR and denitrification would affect performances. Also, the integration of fixed film media into the anoxic reactors affected performances differently from media in aerobic reactors.     

Metabolic versatility in full-scale wastewater treatment plants performing enhanced biological phosphorus removal

This study analysed the enhanced biological phosphorus removal (EBPR) microbial community and metabolic performance of five full-scale EBPR systems by using fluorescence in situ hybridisation combined with off-line batch tests fed with acetate under anaerobic–aerobic conditions. The phosphorus accumulating organisms (PAOs) in all systems were stable and showed little variability between each plant, while glycogen accumulating organisms (GAOs) were present in two of the plants. The metabolic activity of each sludge showed the frequent involvement of the anaerobic tricarboxylic acid cycle (TCA) in PAO metabolism for the anaerobic generation of reducing equivalents, in addition to the more frequently reported glycolysis pathway. Metabolic variability in the use of the two pathways was also observed, between different systems and in the same system over time. The metabolic dynamics was linked to the availability of glycogen, where a higher utilisation of the glycolysis pathway was observed in the two systems employing side-stream hydrolysis, and the TCA cycle was more active in the A²O systems. Full-scale plants that showed higher glycolysis activity also exhibited superior P removal performance, suggesting that promotion of the glycolysis pathway over the TCA cycle could be beneficial towards the optimisation of EBPR systems.     

Limited filamentous bulking in order to enhance integrated nutrient removal and effluent quality

Limited filamentous bulking has been proposed as a means to enhance floc size and make conditions more favorable for simultaneous nitrification/Denitrification (SND). Moreover a slightly heightened SVI is supposed to increase the removal of small particulates in the clarifier. Integrated nitrogen, phosphorus and COD removal performance under limited filamentous bulking was investigated using a bench-scale plug-flow enhanced biological phosphorus removal (EBPR) reactor fed with raw domestic wastewater. Limited filamentous bulking in this study was mainly induced by low DO levels, while other influencing factors associated with filamentous bulking (F/M, nutrients, and wastewater characteristics) were not selective for filamentous bacteria. The optimum scenario for integrated nitrogen, phosphorus and COD removal was achieved under limited filamentous bulking with an SVI level of 170-200 (associated with a DO of 1.0-1.5 mg/L). The removal efficiencies of COD, TP and NH₄⁺-N were 90%, 97% and 92%, respectively. Under these conditions, the solid-liquid separation was practically not affected and sludge loss was never observed. A well-clarified effluent with marginal suspended solids was obtained. The results of this study indicated the feasibility of limited filamentous bulking under low DO as a stimulation of simultaneous nitrification/denitrification for enhancing nutrient removal and effluent quality in an EBPR process.     

The role of the microbial stringent response in excess intracellular accumulation of phosphorous in mixed consortia fed synthetic wastewater

Four bench-scale sequencing batch reactors (SBRs) seeded with activated sludge were operated under either fully oxic or anoxic/oxic conditions and fed synthetic wastewater containing either peptone or acetate. The function of each reactor was assessed through the measure of (i) soluble chemical oxygen demand, orthophosphate, ammonia, and nitrate; and (ii) biomass concentrations of phosphorus, polyhydroxyalkanoate, guanosine tetraphosphate, adenosine monophosphate, adenosine diphosphate, and adenosine triphosphate. In all four reactors, the biomass concentration of phosphorous was correlated statistically with the biomass concentration of ppGpp. The microbial consortia in all four reactors removed an appreciable quantity of phosphorous from solution (67-99%), and the net quantity of phosphorous removed from solution corresponded to the net increase in the biomass concentration of phosphorous. Hence, the microbial stringent response (MSR) was associated with excess intracellular accumulation of phosphorous in mixed microbial consortia fed synthetic wastewater. With recognition of the potential role of the MSR in the removal of soluble phosphorous from wastewater, additional research may lead to further optimization of treatment technologies and the development of new treatment systems for the biological removal of phosphorus from wastewater.     

The nature of the carbon source rules the competition between PAO and denitrifiers in systems for simultaneous biological nitrogen and phosphorus removal

The presence of nitrate in the theoretical anaerobic reactor of a municipal WWTP aiming at simultaneous C, N and P removal usually leads to Enhanced Biological Phosphorus Removal (EBPR) failure due to the competition between PAO and denitrifiers for organic substrate. This problem was studied in a continuous anaerobic-anoxic-aerobic (A²/O) pilot plant (146 L) operating with good removal performance and a PAO-enriched sludge (72%). Nitrate presence in the initially anaerobic reactor was studied by switching the operation of the plant to an anoxic-aerobic configuration. When the influent COD composition was a mixture of different carbon sources (acetic acid, propionic acid and sucrose) the system was surprisingly able to maintain EBPR, even with internal recycle ratios up to ten times the influent flow rate and COD limiting conditions. However, the utilisation of sucrose as sole carbon source resulted in a fast EBPR failure. Batch tests with different nitrate concentrations (0-40 mg L⁻¹) were performed in order to gain insight into the competition for the carbon source in terms of P-release or denitrification rates and P-release/C-uptake ratio. Surprisingly, no inhibitory or detrimental effect on EBPR performance due to nitrate was observed. A model based on ASM2d but considering two step nitrification and denitrification was developed and experimentally validated. Simulation studies showed that anaerobic VFA availability is critical to maintain EBPR activity.     

Meeting the phosphorus consent with biological nutrient removal under UK winter conditions

It is estimated that up to 342 wastewater treatment plants (Wwtps) in England and Wales will require a phosphorus (P) consent by 2010. Although biological P removal is considered to be the most sustainable option for P removal, it has always been problematic for plants that remove both nitrogen and P due to the inadequate concentration of organic material during wet periods. Two biological nutrient removal (BNR) configurations, the Johannesburg (JHB) process and a combined JHB and five-stage Bardenpho process, were evaluated over a period of 2 years to assess the impact of sewage strength on bio-P removal. The JHB achieved an average effluent total phosphorus (TP) of 2.4 mg/L and the combined JHB and five-stage process averaged 1.4 mg/L effluent TP. The major problems affecting the performance of both configurations were: dissolved oxygen (DO) in the recycled mixed liquor, nitrate in the return activated sludge (RAS) and low influent biological oxygen demand (BOD) concentrations. Acetate dosing proved successful as a source of volatile fatty acids (VFAs) in the anaerobic zone during periods of low-strength sewage. An acetate dosing strategy based on the influent flow rate to the plant was found to be a simple and effective technique that ensured that a consent of <1 mg TP/L could be met.     

Effect of nitrite on phosphate uptake by phosphate accumulating organisms

In biological nitrogen removal processes, nitrite can be formed and accumulated through both nitrification and denitrification. Despite the fact that, in practice, biological phosphate removal (BPR) is often combined with biological nitrogen removal, there are only a few publications reporting the effect of nitrite on BPR. In this study, phosphate-accumulating organisms (PAO) were cultivated in an anaerobic-anoxic-aerobic sequencing batch reactor (SBR). The effect of nitrite on the enrichment of the sludge with PAO, the phosphate uptake rates and the sludge respiration was investigated. The results indicate that (1) presence of nitrite inhibits both aerobic and anoxic (denitrifying) phosphate uptake, (2) aerobic phosphate uptake was more affected than anoxic phosphate uptake, (3) presence of nitrite could be one of the factors enhancing the presence of glycogen accumulating organisms (GAO)--competitors to PAO for substrate in the anaerobic phase, and (4) it is required to monitor and control nitrite accumulation in a full-scale wastewater treatment plants.     

Kinetic competition between phosphorus release and denitrification on sludge under anoxic condition

The kinetic behaviors of simultaneous phosphorus release and denitrification on sludge were investigated under anoxic condition. A phosphorus enriched sludge produced from Anaerobic-Anoxic-Oxic (AnAO) process under various SRT (5, 10 and 15 days) operation conditions was carried out in a series of batch tests. Experimental results indicated that the available organic substrate determined the kinetic behaviors of phosphorus release/uptake and denitrification. The simultaneous phosphorus release and denitrification demonstrated a kinetic competition under anoxic conditions in the presence of an available organic substrate. When the substrate was abundant, sludge was under “releasable-phosphorus-limited” condition; phosphorus release rate decreased slightly by nitrate inhibition. However, nitrate significantly inhibited phosphorus release when sludge was under “initial-substrate-limited” condition. Moreover, the sludge's phosphorus contents (as created by different SRT processes) dominated the kinetics of competition between phosphorus release and denitrification. The sludge with a high phosphorus content had a higher phosphorus release rate in accordance with a lower denitrification rate. Additionally, the substrate sequestrated rate markedly increased under the condition of simultaneous phosphorus release and denitrification. Finally, a preliminary metabolism model of denitrifying phosphorus removal bacteria was proposed, and found to be capable of adequately accounting for simultaneous phosphorus release and denitrification under anoxic conditions.