Heterogeneous diffusion in aerobic granular sludge

Aerobic granular sludge (AGS) technology allows simultaneous nitrogen, phosphorus, and carbon removal in compact wastewater treatment processes. To operate, design, and model AGS reactors, it is essential to properly understand the diffusive transport within the granules. In this study, diffusive mass transfer within full-scale and lab-scale AGS was characterized with nuclear magnetic resonance (NMR) methods. Self-diffusion coefficients of water inside the granules were determined with pulsed-field gradient NMR, while the granule structure was visualized with NMR imaging. A reaction-diffusion granule-scale model was set up to evaluate the impact of heterogeneous diffusion on granule performance. The self-diffusion coefficient of water in AGS was ∼70% of the self-diffusion coefficient of free water. There was no significant difference between self-diffusion in AGS from full-scale treatment plants and from lab-scale reactors. The results of the model showed that diffusional heterogeneity did not lead to a major change of flux into the granule (<1%). This study shows that differences between granular sludges and heterogeneity within granules have little impact on the kinetic properties of AGS. Thus, a relatively simple approach is sufficient to describe mass transport by diffusion into the granules.     

Degrading high-strength phenol using aerobic granular sludge

Aerobic granules were adopted to degrade high-strength phenol wastewater in batch experiments. The acclimated granules effectively degraded phenol at a concentration of up to 5,000 mg l⁻¹ without severe inhibitory effects. The biodegradation of phenol by activated sludge was inhibited at phenol concentrations >3,000 mg l⁻¹. The granules were composed of cells embedded in a compact extracellular matrix. After acid or alkaline pretreatment, the granules continued to degrade phenol at an acceptable rate. The polymerase chain reaction-denaturing gradient gel electrophoresis technique was employed to monitor the microbial communities of the activated sludge and the aerobic granules following their being used to treat high concentrations of phenol in batch tests.     

Microbial diversity differences within aerobic granular sludge and activated sludge flocs

In this study, we investigated during 400 days the microbial community variations as observed from 16S DNA gene DGGE banding patterns from an aerobic granular sludge pilot plant as well as the from a full-scale activated sludge treatment plant in Epe, the Netherlands. Both plants obtained the same wastewater and had the same relative hydraulic variations and run stable over time. For the total bacterial population, a similarity analysis was conducted showing that the community composition of both sludge types was very dissimilar. Despite this difference, general bacterial population of both systems had on average comparable species richness, entropy, and evenness, suggesting that different bacteria were sharing the same functionality. Moreover, multi-dimensional scaling analysis revealed that the microbial populations of the flocculent sludge system moved closely around the initial population, whereas the bacterial population in the aerobic granular sludge moved away from its initial population representing a permanent change. In addition, the ammonium-oxidizing community of both sludge systems was studied in detail showing more unevenness than the general bacterial community. Nitrosomonas was the dominant AOB in flocculent sludge, whereas in granular sludge, Nitrosomonas and Nitrosospira were present in equal amounts. A correlation analysis of process data and microbial data from DGGE gels showed that the microbial diversity shift in ammonium-oxidizing bacteria clearly correlated with fluctuations in temperature.     

PPCPs removal by aerobic granular sludge membrane bioreactor

An aerobic granular sludge membrane bioreactor (GMBR) was applied to the treatment of pharmaceutical and personal care products (PPCPs) wastewater. The influence of granular sludge on five antibiotic and antiphlogistic PPCPs wastewater and the removal effect of methyl alcohol and conventional organic matter were investigated while constantly reducing the density of inflow organic matter. The results showed that the sludge granulation process in the system was rapid but unstable, and that the system exhibits a dissolution–reunion dynamic equilibrium. The reactor demonstrated varying removal effects of PPCPs on different objects. The use of a GMBR was more effective for the removal of prednisolone, naproxen, and ibuprofen; the first two drugs were lower the average removal rate of which reached 98.46 and 84.02 %, respectively; whereas the average removal rate of ibuprofen was 63.32 %. By contrast, the GMBR has an insignificant degradation effect on antibiotics such as amoxicillin, indicating that such antibiotic medicine is not easily degraded by microorganisms, which plays different roles in system operation. Because of the different chemical structures and characteristics of drugs that result in various degradation behavior. During the GMBR granulation process, the value of mixed liquor volatility suspended solids (MLVSS) gradually increases from 1.5 to 4.1 g/L during the GMBR granulation process, and the removal rate of COD_Cr reaches up to 87.98 %. After reducing the density of organic matter is reduced, the removal rates of NH₃-N and TP both reach more than 90 %, respectively. Moreover, the proposed technique is considerably effective in the removal of methanol.     

Evolution of bioaggregate strength during aerobic granular sludge formation

This work investigated the modification of aggregate properties during the formation of granular sludge in a sequencing batch airlift reactor (SBAR). The cohesion of biological aggregates was quantified by subjecting sludge samples to two different controlled shear stresses in a stirred reactor. For reference sludge (without granules), flocs broke and reformed easily, indicating that floc size was controlled by the turbulence micro-scale (Kolmogorov scale, here from 17 μm to 62 μm). In contrast, granules showed high strength which enabled them to resist turbulence and their size was no longer imposed by the Kolmogorov micro-scale. Different steps were observed during the granulation process: a first increase of aggregate cohesion associated with a decrease in sludge volume index (SVI), a growth of aggregates with detachment of fragile particles from the surface and, finally, an increase in the sizes of small and large granules to reach a pseudo-stable size distribution. Results suggest that small particles could have formed the seeds for new granules, as they were maintained in the bioreactor. Here, granular sludge was formed in an SBAR with a conventional settling time (30 min), i.e. without particle washout, and with a low superficial air velocity (SAV = 0.6 cm s⁻¹): it is thus demonstrated that high SAV and low settling time are not necessary to produce granules, but probably only accelerate the accumulation of granules. It is shown that the increase of cohesion is the initial phenomenon explaining the granule formation concomitantly with bacterial aggregates densification. It seems important, in the future, to investigate the reasons for this cohesion increase, which is possibly explained either by bacterial bounding interactions or the excretion of extracellular polymeric substances (EPS).     

Mathematical modeling of aerobic granular sludge: A review

Aerobic granulation may play an important role in the field of wastewater treatment due to the advantages of aerobic granules compared to the conventional sludge flocs, such as denser structure, better settleability and ensured solid-effluent separation, higher biomass concentration, and greater ability to withstand shock loadings, which is promising for a full-scale implementation. As an aid for this implementation, mathematical modeling would be an invaluable tool. In this paper, the existing mathematical models available in literature concerning aerobic granule systems are reviewed, including the modeling of the dynamic facets of the aerobic granulation process, the mass transfer and detachment in aerobic granules, the granule-based sequencing batch reactor, the fate of microbial products in granules, and the multi-scale modeling of aerobic granular sludge. An overview of the parameters used in the aerobic granular modeling approaches is also presented. Our growing knowledge on mathematical modeling of aerobic granule might facilitate the engineering and optimization of aerobic granular sludge technology as one of the most promising techniques in the biological wastewater treatment.     

Improved phosphate removal by selective sludge discharge in aerobic granular sludge reactors

Two lab-scale aerobic granular sludge sequencing batch reactors were operated at 20 and 30°C and compared for phosphorus (P) removal efficiency and microbial community composition. P-removal efficiency was higher at 20°C (>90%) than at 30°C (60%) when the sludge retention time (SRT) was controlled at 30 days by removing excess sludge equally throughout the sludge bed. Samples analyzed by fluorescent in situ hybridization (FISH) indicated a segregation of biomass over the sludge bed: in the upper part, Candidatus Competibacter phosphatis (glycogen-accumulating organisms--GAOs) were dominant while in the bottom, Candidatus Accumulibacter phosphatis (polyphosphate-accumulating organisms--PAOs) dominated. In order to favour PAOs over GAOs and hence improve P-removal at 30°C, the SRT was controlled by discharging biomass mainly from the top of the sludge bed (80% of the excess sludge), while bottom granules were removed in minor proportions (20% of the excess sludge). With the selective sludge removal proposed, 100% P-removal efficiency was obtained in the reactor operated at 30°C. In the meantime, the biomass in the 30°C reactor changed in color from brownish-black to white. Big white granules appeared in this system and were completely dominated by PAOs (more than 90% of the microbial population), showing relatively high ash content compared to other granules. In the reactor operated at 20°C, P-removal efficiency remained stable above 90% regardless of the sludge removal procedure for SRT control. The results obtained in this study stress the importance of sludge discharge mainly from the top as well as in minor proportions from the bottom of the sludge bed to control the SRT in order to prevent significant growth of GAOs and remove enough accumulated P from the system, particularly at high temperatures (e.g., 30°C).     

Investigation on the formation and kinetics of glucose-fed aerobic granular sludge

Aerobic granular sludge was cultivated in a glass sequencing batch reactor (SBR) with glucose synthetic wastewater. The spherical shaped granules were observed on 4th day with the mean diameter of 0.1 mm. With the increase of chemical oxygen demand (COD) concentration of the influent, aerobic granules grew matured, the size of which ranged from 1.2 to 1.9 mm. The aerobic granular sludge could sustain high organic loading rate (about 4.0 g COD L⁻¹ d⁻¹), with good settling ability (settling velocity 36 m/h) and high biomass concentration (MLSS 6.7 ±0.2 g/L). Experimental data indicated that the substrate utilization and biomass growth kinetics followed Monod's kinetics model approximately. The corresponding kinetic coefficients of maximum specific substrate utilization rate (k), half velocity coefficient (K_s), growth yield coefficient (Y) and decay coefficient (K_d) were 13.2 d⁻¹, 275.8 mg/L, 0.183–0.250 mg MLSS/mg COD and 0.023–0.075 d⁻¹, respectively, which made aerobic granules have short setup period, high rate of substrate utilization and little surplus sludge.     

Investigation on the properties and kinetics of glucose-fed aerobic granular sludge

Aerobic granulation is a process in which suspended biomass aggregate and form discrete well-defined granules in aerobic systems. To investigate the properties and kinetics of aerobic granular sludge, aerobic granules were cultivated with glucose synthetic wastewater in a series of sequencing batch reactors (SBR). The spherical shaped granules were observed on 8th day with the mean diameter of 0.1 mm. With the organic loading rate (OLR) being increased to 4.0 g COD L⁻¹ d⁻¹, aerobic granules grew matured with spherical shape. The size of granules ranged from 1.2 to 1.8 mm, and the corresponding settling velocity of individual granule was 24.2–36.4 m h⁻¹. The oxygen utilization rate (OUR) of mature granules was 41.90 g O₂ kg MLSS⁻¹ h⁻¹, which was two times higher than that of activated sludge (18.32 g O₂ kg MLSS⁻¹ h⁻¹). The experimental data indicated that the substrate utilization and biomass growth kinetics generally followed Monod's kinetics model. The corresponding kinetic coefficients of k (maximum specific substrate utilization rate), K_s (half velocity coefficient), Y (growth yield coefficient) and K_d (decay coefficient) were determined as follows, k_c = 23.65 d⁻¹, K_c = 3367.05 mg L⁻¹, K_N = 0.038 d⁻¹, K_N = 29.65 mg L⁻¹, Y = 0.1927–0.2022 mg MMLS (mg COD)⁻¹ and K_d = 0.00845–0.0135 d⁻¹, respectively. Those properties of aerobic granules made aerobic granules system had a short setup period, high substrate utilization rate and low sludge production.     

Influence of phenol on cultures of acetate-fed aerobic granular sludge

AIMS: This paper attempts to investigate the inhibition of phenol on the acetate utilization in acetate-fed aerobic granular sludge culture. METHODS AND RESULTS: Acetate-fed aerobic granules with a mean diameter of 1.0 mm were predeveloped in a column sequencing aerobic sludge blanket reactor. The present study looked into the utilization kinetics of acetate by acetate-fed aerobic granules in the presence of different phenol concentrations ranging from 0 mg l(-1) to 50 mg l(-1). For this purpose, batch experiments were conducted at 25 degrees C, while the initial biomass and acetate concentrations were in a range of 109-186 mg mixed liquor suspended solids (MLSS) l(-1) and 185-300 mg acetate-chemical oxygen demand (COD) l(-1). Results showed that the utilization of acetate in the presence of phenol was subject to a zero-order reaction kinetics. The relative phenol concentration in terms of the ratio of initial phenol concentration (C(p)) to initial biomass concentration (X(0)) was used to describe the real inhibitory strength of phenol imposed on acetate-fed aerobic granules. When the C(p)/X(0) ratio increased from 0 to 0.19 mg phenol mg(-1) MLSS, the zero-order reaction rate constant of acetate dropped from 1.15 mg l(-1) min(-1) to 0.38 mg l(-1) min(-1), and a similar trend was also observed in specific oxygen utilization rate. As compared to the control test without addition of phenol, the acetate-COD removal efficiency was reduced by nearly 50% at a C(p)/X(0) value of 0.19 mg phenol mg(-1) MLSS. It was found that biodegradation of phenol was negligible in acetate-fed aerobic granular sludge batch culture. CONCLUSIONS: It appears that phenol can seriously repress the utilization of acetate in the acetate-fed aerobic granular sludge batch cultures. A simple zero-order reaction model could adequately describe the utilization of acetate by acetate-fed aerobic granules in the presence of phenol. SIGNIFICANCE AND IMPACT OF THE STUDY: It is expected that this study would lead to a better understanding of the behaviour of acetate-fed aerobic granules in the presence of inhibitory organic compounds.     

好氧颗粒污泥对不同底物组成的响应

在序批式间歇反应器（R1、R2和R3）中,采用乙酸钠（R1）、蔗糖（R2）和苯酚（R3）三种不同基质作为碳源,均成功地培养出了好氧颗粒污泥;考察了不同颗粒污泥的理化性质及其对污染物的转化能力。结果表明,R1中颗粒污泥外观呈黄色,其主要的微生物菌群为细菌;R2中颗粒污泥外观呈黑色,内部含有丝状菌;而R3中颗粒污泥表面被大量丝状菌包裹,颗粒污泥呈淡黄色。在进水COD1000mg／L时R1、R2和R3中颗粒污泥比有机物的利用速率大小顺序为R3〉R1〉R2,而COD的去除率顺序却为R2〉R1〉R3。在进水氨氮40mg／L时,R1、R2和R3中氨氮的去除率分别在91％、96％和80％以上。以不同的底物培养出不同的好氧颗粒污泥可以拓展其在有毒化学物质如酚类化合物和高浓度工业废水生物处理中的应用。     

A generalized model for settling velocity of aerobic granular sludge

Aerobic granulation is a novel biotechnology recently receiving intensive research attention. Aerobic granules developed in SBR can be as big as several millimeters, thus the traditional models describing the settling velocity of activated sludge are no long valid in aerobic granules culture. In this study, a new type of model was developed for the settling velocity of aerobic granules. This model shows that the settling velocity of aerobic granules is the function of SVI, mean size of granules and biomass concentration of granules. When the size of bioparticle is small enough, the proposed model reduces to the well-known Vesilind equation. Results indicated that the proposed model could satisfactorily fit experimental results obtained in the course of aerobic granulation under different conditions, while the Vesilind equation failed to or very poorly fit the experimental data. In addition, the proposed model can also be extended to anaerobic granules. The settling velocity is one of the most important parameters in both aerobic and anaerobic granulation, and successful biogranulation is highly related to the manipulation of settling velocity. It was demonstrated that the proposed model can sever as a useful tool for design and operation engineers to properly select the settling velocity for enhanced aerobic and anaerobic granulation.     

Characterization of aerobic granular sludge at various organic loading rates

Aerobic granular sludge was cultivated from activated sludge with two types of supports, namely bivalve shell carrier (BSC) and anaerobic granules (ANG). Granules were characterized at different organic loading rates (OLRs) ranging from 2.5 to 15 kg COD/m³ d and these granules were observed to withstand high OLRs. The physico-chemical characteristics of the aerobic granules were better than those of seed sludge. The granule formation with ANG support was found to be similar to that of non-support cultivation, i.e. formation from activated sludge only. By contrast, BSC support showed better performance in terms of faster settleability, compactness and especially resistance against organic shock loading. It also enabled self-cleaning effect by removing biofilm attached on the reactor wall during the start-up phase resulting rapid granulation process.     

Effect of pH on nickel biosorption by aerobic granular sludge

The Ni2+ biosorption by aerobic granular sludge was studied at various initial pH values of 2-7. Results showed that the initial pH would play an important role in the Ni2+ removal by aerobic granules and affected the zeta potential of aerobic granules. A thermodynamic equilibrium isotherm previously developed can fit the experimental data very well at all studied pH values. The close relationship between the zeta potential and Ni2+ biosorption capacity of aerobic granules showed the electrostatic attraction between the aerobic granules and Ni2+ ions. It was also found that some light metals, such as K+, Mg2+ and Ca2+ would be released into the bulk solution during the Ni2+ uptake onto the aerobic granules, which in turn indicated that ion-exchange was one of the Ni2+ biosorption mechanisms.     

Organics and nitrogen removal and sludge stability in aerobic granular sludge membrane bioreactor

Novel aerobic granular sludge membrane bioreactor (GMBR) was established by combining aerobic granular sludge technology with membrane bioreactor (MBR). GMBR showed good organics removal and simultaneous nitrification and denitrification (SND) performances for synthesized wastewater. When influent total organic carbon (TOC) was 56.8-132.6 mg/L, the TOC removal of GMBR was 84.7-91.9%. When influent ammonia nitrogen was 28.1-38.4 mg/L, the ammonia nitrogen removal was 85.4-99.7%, and the total nitrogen removal was 41.7-78.4%. Moreover, batch experiments of sludge with different particle size demonstrated that: (1) flocculent sludge under aerobic condition almost have no denitrification capacity, (2) SND capacity was caused by the granular sludge, and (3) the denitrification rate and total nitrogen removal efficiency were enhanced with the increased particle size. In addition, study on the sludge morphology stability in GMBR showed that, although some granular sludge larger than 0.9 mm disaggregated at the beginning of operation, the granular sludge was able to maintain the stability of its granular morphology, and at the end of operation, the amount of granular sludge (larger than 0.18 mm) stabilized in GMBR was more than 56-62% of the total sludge concentration. The partial disaggregation of large granules is closely associated with the change of operating mode from sequencing batch reactor (SBR) system to MBR system.     

Proteolytic activity in stored aerobic granular sludge and structural integrity

Aerobic granules lose stability during storage. The goal of this work was to highlight the main cause of stability loss for stored granules as intracellular protein hydrolysis. The quantity of extracellular proteins was noted to be significantly lower during granule storage, and protease enzyme activities were correspondingly higher in the cores of stored granules. The proteolytic bacteria, which secrete highly active protease enzymes, were for the first time isolated and characterized by analyzing 16S rDNA sequences. The proteolytic bacteria belonged to the genera Pseudomonas, Raoultella, Acinetobacter, Pandoraea, Klebsiella, Bacillus and uncultured bacterium, and were grouped into Proteobacteria, Enterobacteria and Firmicutes. The PB1 (Pseudomonas aeruginosa) strain, which exhibited very high proteolytic activity during the skim milk agar test, was located at the core regime with active protease enzymes, and was close to the obligate anaerobic strain Bacteroides sp. Hence, the extracellular proteins in stored granules were proposed to be hydrolyzed by enzymes secreted by proteolytic bacteria with the hydrolyzed products ultimately being used by nearby anaerobic strains. This process gradually digests the protein core, and eventually consumes the entire granule.     

Variable aeration in sequencing batch reactor with aerobic granular sludge

This study investigated the effects of reduced aeration in famine period on the performance of sequencing batch reactor (SBR) with aerobic granular sludge. Aerobic granules were first cultivated in two SBRs (R1 and R2) with acetate as sole carbon source. From operation day 27, aeration rate in R1 was reduced from 1.66 to 0.55 cm s(-1) from 110 min to the end of each cycle and further reduced from 30 min to the end of each cycle from day 63. R2 as a control was operated with a constant aeration rate of 1.66 cm s(-1) in the whole cycle during the entire experimental period. Results showed that changing trends of SVI, concentration, average size and VSS/SS of biomass with time in R1 and R2 were similar although different aeration modes were adopted. At steady state, SVI of aerobic granules and biomass concentration maintained at about 40 ml g(-1) and 6 g l(-1), respectively. Average size of granules was about 750 microm in R1 while 550 microm in R2. This is the first study to demonstrate that aerobic granular sludge could be stable at reduced aeration rate in famine period during more than 3-month operation. Such an operation strategy with reduced aeration rate will lead to a significant reduction of energy consumption, which makes the aerobic granular sludge technology more competitive over conventional activated sludge process. Furthermore, the stability of aerobic granular system with variable aeration further indicates that the difference of physiology and kinetics of aerobic granule in feast and famine periods results in the different requirements of oxygen and shear stress for the stability of granules, which will deepen the understanding of mechanism of aerobic granulation in sequencing batch reactor.     

Reactivation characteristics of stored aerobic granular sludge using different operational strategies

Aerobic granules after 6 months storage were employed in identical sequencing batch reactors (SBRs) using synthetic wastewater to investigate the impacts of different operational strategies on granules' reactivation process. The SBRs were operated under three operational strategies for reactivation of (a) different organic loading rate (OLR); (b) different ammonia concentration; and (c) different shear force (a superficial upflow air velocity). The results indicated that granules after long-term storage could be successfully recovered after 7 days of operation, and the excellent granule reactivation performance was closely related to the operational strategies, since inappropriate operational strategies could cause the outgrowth of filamentous bacteria and granule disintegration. Based on comprehensive comparison of reactivation performance under different operational strategies, the optimal operation strategy for granule reactivation was suggested at OLR of 0.8 kg COD/m(3)/day, ammonia concentration of 15-20 mg/L, and a superficial upflow air velocity of 2.6 cm/s. After 7 days operation under the optimal strategy, the dark brown granules (12 months storage) restored their bioactivities to previous state, in terms of COD removal efficiency (97.44%) and specific oxygen uptake rate (40.63 mg O(2)/g SS h(-1)). The results shed light on the future practical application of stored aerobic granules as bioseed for reactor fast start-up.     

Integration of aerobic granular sludge and membrane bioreactors for wastewater treatment

Environmental deterioration together with the need for water reuse and the increasingly restrictive legislation of water quality standards have led to a demand for compact, efficient and less energy consuming technologies for wastewater treatment. Aerobic granular sludge and membrane bioreactors (MBRs) are two technologies with several advantages, such as small footprint, high-microbial density and activity, ability to operate at high organic- and nitrogen-loading rates, and tolerance to toxicity. However, they also have some disadvantages. The aerobic granular sludge process generally requires post-treatment in order to fulfill effluent standards and MBRs suffer from fouling of the membranes. Integrating the two technologies could be a way of combining the advantages and addressing the main problems associated with both processes. The use of membranes to separate the aerobic granules from the treated water would ensure high-quality effluents suitable for reuse. Moreover, the use of granular sludge in MBRs has been shown to reduce fouling. Several recent studies have shown that the aerobic granular membrane bioreactor (AGMBR) is a promising hybrid process with many attractive features. However, major challenges that have to be addressed include how to achieve granulation and maintain granular stability during continuous operation of reactors. This paper aims to review the current state of research on AGMBR technology while drawing attention to relevant findings and highlight current limitations.     

Achieving high-rate partial nitritation with aerobic granular sludge at low temperatures

Biodegradation - Partial nitritation is necessary for the implementation of the mainstream anammox (anaerobic ammonium oxidation) process in wastewater treatment plants. However, the difficulty in...