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Aerobic granular sludge (AGS) and conventional activated sludge (CAS) are two different biological wastewater treatment processes. AGS consists of self-immobilised microorganisms that are transformed into spherical biofilms, whereas CAS has floccular sludge of lower density. In this study, we investigated the treatment performance and microbiome dynamics of two full-scale AGS reactors and a parallel CAS system at a municipal WWTP in Sweden. Both systems produced low effluent concentrations, with some fluctuations in phosphate and nitrate mainly due to variations in organic substrate availability. The microbial diversity was slightly higher in the AGS, with different dynamics in the microbiome over time. Seasonal periodicity was observed in both sludge types, with a larger shift in the CAS microbiome compared to the AGS. Groups important for reactor function, such as ammonia-oxidising bacteria (AOB), nitrite-oxidising bacteria (NOB), polyphosphate-accumulating organisms (PAOs) and glycogen-accumulating organisms (GAOs), followed similar trends in both systems, with higher relative abundances of PAOs and GAOs in the AGS. However, microbial composition and dynamics differed between the two systems at the genus level. For instance, among PAOs, Tetrasphaera was more prevalent in the AGS, while Dechloromonas was more common in the CAS. Among NOB, Ca. Nitrotoga had a higher relative abundance in the AGS, while Nitrospira was the main nitrifier in the CAS. Furthermore, network analysis revealed the clustering of the various genera within the guilds to modules with different temporal patterns, suggesting functional redundancy in both AGS and CAS. Key points: • Microbial community succession in parallel full-scale aerobic granular sludge (AGS) and conventional activated sludge (CAS) processes. • Higher periodicity in microbial community structure in CAS compared to in AGS. • Similar functional groups between AGS and CAS but different composition and dynamics at genus level. [ABSTRACT FROM AUTHOR]

Complex substrates, like proteins, carbohydrates, and lipids, are major components of domestic wastewater, and yet their degradation in biofilm-based wastewater treatment technologies, such as aerobic granular sludge (AGS), is not well understood. Hydrolysis is considered the rate-limiting step in the bioconversion of complex substrates, and as such, it will impact the utilization of a large wastewater COD (chemical oxygen demand) fraction by the biofilms or granules. To study the hydrolysis of complex substrates within these types of biomass, this paper investigates the anaerobic activity of major hydrolytic enzymes in the different sludge fractions of a full-scale AGS reactor. Chromogenic substrates were used under fully mixed anaerobic conditions to determine lipase, protease, α-glucosidase, and β-glucosidase activities in large granules (>1 mm in diameter), small granules (0.2–1 mm), flocculent sludge (0.045–0.2 mm), and bulk liquid. Furthermore, composition and hydrolytic activity of influent wastewater samples were determined. Our results showed an overcapacity of the sludge to hydrolyze wastewater soluble and colloidal polymeric substrates. The highest specific hydrolytic activity was associated with the flocculent sludge fraction (1.5–7.5 times that of large and smaller granules), in agreement with its large available surface area. However, the biomass in the full-scale reactor consisted of 84% large granules, making the large granules account for 55–68% of the total hydrolytic activity potential in the reactor. These observations shine a new light on the contribution of large granules to the conversion of polymeric COD and suggest that large granules can hydrolyze a significant amount of this influent fraction. The anaerobic removal of polymeric soluble and colloidal substrates could clarify the stable granule formation that is observed in full-scale installations, even when those are fed with complex wastewaters. Key points: • Large and small granules contain >70% of the hydrolysis potential in an AGS reactor. • Flocculent sludge has high hydrolytic activity but constitutes <10% VS in AGS. • AGS has an overcapacity to hydrolyze complex substrates in domestic wastewater. [ABSTRACT FROM AUTHOR]

Aerobic granular sludge (AGS) is a microbial biofilm self-aggregation, which is effective for nutrient and pollutant removal, through the development of dense microbial layers bound together with extracellular polymeric substances (EPSs). However, long start-up times and granule disintegration are still challenges ahead. An array of external additives, including ion chelating agents, sludge-based enhancers, and magnetic influence have been tested to overcome these barriers. The application of such additives may promote enhanced EPS production, neutralization of charges on the bacterial surface, acts as a core-induced agent, or as a bridge to connect EPSs and cell surfaces. Although additives may improve the granule formation without reducing treatment efficiencies, there are still environmental concerns due to the fate and toxicity of discharged excess sludge. This mini-review identifies an array of external additives and their mechanisms to improve granulation properties, and proposes discussion about the technical and economic viability of these additives. Key points: • Additives reduce granulation time and repair granule disintegration. • Biopolymer-based additives fulfill technical and environmental requirements. • Sludge-based additives are cheap and in line with the resource recovery concept. • The need for environmental-friendly additives for aerobic granular sludge process. • External additives affect granular biomass size distribution. [ABSTRACT FROM AUTHOR]

Granular activated sludge has gained increasing interest due to its potential in treating wastewater in a compact and efficient way. It is well-established that activated sludge can form granules under certain environmental conditions such as batch-wise operation with feast-famine feeding, high hydrodynamic shear forces, and short settling time which select for dense microbial aggregates. Aerobic granules with stable structure and functionality have been obtained with a range of different wastewaters seeded with different sources of sludge at different operational conditions, but the microbial communities developed differed substantially. In spite of this, granule instability occurs. In this review, the available literature on the mechanisms involved in granulation and how it affects the effluent quality is assessed with special attention given to the microbial interactions involved. To be able to optimize the process further, more knowledge is needed regarding the influence of microbial communities and their metabolism on granule stability and functionality. Studies performed at conditions similar to full-scale such as fluctuation in organic loading rate, hydrodynamic conditions, temperature, incoming particles, and feed water microorganisms need further investigations. [ABSTRACT FROM AUTHOR]

Aerobic granular sludge (AGS), a self-immobilized microbial consortium containing different functional microorganisms, is receiving growing attention, since it has shown great technological and economical potentials in the field of wastewater treatment. Microbial community is crucial for the formation, stability, and pollutant removal efficiency of aerobic granules. This mini-review systematically summarizes the recent findings of the microbial community structure and function of AGS and discusses the new research progress in the microbial community dynamics during the granulation process and spatial distribution patterns of the microbiota in AGS. The presented information may be helpful for the in-depth theoretical study and practical application of AGS technology in the future. [ABSTRACT FROM AUTHOR]

With rapid industrialization and urbanization in the nineteenth century, the activated sludge process (ASP) has experienced significant steps forward in the face of greater awareness of and sensitivity toward water-related environmental problems. Compared with conventional flocculent ASP, the major advantages of granular sludge are characterized by space saving and resource recovery, where the methane and hydrogen recovery in anaerobic granular and 50% more space saving, 30-50% of energy consumption reduction, 75% of footprint cutting, and even alginate recovery in aerobic granular. Numerous engineers and scientists have made great efforts to explore the superiority over the last 40 years. Therefore, a bibliometric analysis was desired to trace the global trends of granular sludge research from 1992 to 2016 indexed in the SCI-EXPANDED. Articles were published in 276 journals across 44 subject categories spanning 1420 institutes across 68 countries. Bioresource Technology (293, 11.9%), Water Research (235, 9.6%), and Applied Microbiology and Biotechnology (127, 5.2%) dominated in top three journals. The Engineering (991, 40.3%), China (906, 36.9%), and Harbin Inst Technol, China (114, 4.6%) were the most productive subject category, country, and institution, respectively. The hotspot is the emerging techniques depended on granular reactors in response to the desired removal requirements and bio-energy production (primarily in anaerobic granular sludge). In view of advanced and novel bio-analytical methods, the characteristics, functions, and mechanisms for microbial granular were further revealed in improving and innovating the granulation techniques. Therefore, a promising technique armed with strengthened treatment efficiency and efficient resource and bio-energy recovery can be achieved. [ABSTRACT FROM AUTHOR]

Bacteria can coordinate and synchronize activities through a cell density-dependent regulatory mechanism called quorum sensing (QS). Bacteria can measure their population by the synthesis, secretion, and perception of QS signal molecules to regulate specific gene expression when the population reaches a critical threshold. QS participates in various microbial processes such as marine organism bioluminescence, bacterial biofilm formation, and virulence factor expression. The use of QS systems mediated by N-acyl-homoserine lactones and autoinducer-2 has been recently recognized as a promising regulatory approach in environmental science and technology that can intrinsically promote the profound comprehension of wastewater treatment from a microbiology perspective. This article reviewed the study of QS in several environmental systems in wastewater treatment, including systems of aerobic granular sludge, biological nitrogen removal, and bioaugmentation, while several future prospects and suggestions are proposed on the basis of current studies. [ABSTRACT FROM AUTHOR]

In this study, we demonstrated that anaerobic granular sludge could be successfully transformed into aerobic granular sludge in a continuous up-flow reactor in 45 days. An aerobic microbial community successfully developed in the granules and high organic matter and nitrogen removal performance was achieved. Under an ammonia nitrogen loading rate of 0.8 kg N/(m day), ammonia nitrogen and the total nitrogen removal efficiency of the reactor reached up to 100 and 93%, respectively. An obvious bacterial community shift in granular sludge was observed during the transformation process. By comparing with the bacterial community in aerobic granules cultivated from floccular activated sludge, some bacteria (affiliated with Comamonadaceae, Xanthomonadaceae, Rhodocyclaceae, Moraxellaceae, and Nitrosomonadaceae) playing significant roles in maintaining the structures and functions of aerobic granules were identified. After the transformation, the granules could be clearly separated into the inner core and outer shell. 16S rRNA gene sequencing results indicated many bacterial species present in both the inner core and outer shell; however, their abundance differed significantly. Overall, this study confirms the feasibility of transforming anaerobic granules into aerobic granules and provides novel approaches and insights to understand the microbial ecology in granular sludge. [ABSTRACT FROM AUTHOR]

In this study, a sequencing batch reactor (SBR), treating synthetic wastewater (COD/ N = 5), was operated in two stages. During stage I, an aeration control strategy based on oxygen uptake rate (OUR) was applied, to accomplish nitrogen removal via nitrite >80%. In stage II, the development of aerobic granular sludge (AGS) was examined while two aeration control strategies (OUR and pH slope) maintained the nitrite pathway and optimized the simultaneous nitrification-denitrification (SND) performance. Stimulation of slow-growing organisms, (denitrifying) polyphosphate-accumulating organisms (D)PAO and (denitrifying) glycogen-accumulating organisms (D)GAO leads to full granulation (at day 200, SVI = 47.0 mL/g and SVI = 43.1 mL/g). The average biological nutrient removal efficiencies, for nitrogen and phosphorus, were 94.6 and 83.7%, respectively. Furthermore, the benefits of an increased dissolved oxygen concentration (1.0-2.0 mg O/L) were shown as biomass concentrations increased with approximately 2 g/L, and specific ammonium removal rate and phosphorus uptake rate increased with 33 and 44%, respectively. It was shown that the combination of both aeration phase-length control strategies provided an innovative method to achieve SND via nitrite in AGS. [ABSTRACT FROM AUTHOR]

The aim of this study was to develop a simple operation strategy for the cultivation of partial nitrification granules (PNGs) treating an autotrophic medium. For this strategy, aerobic granular sludge adapted to high concentration organics removal was seeded in a sequencing batch reactor (SBR) with a height/diameter ratio of 3.8, and the ratio of organics to the ammonia nitrogen-loading rate (C/N ratio) in the influent was employed as the main control parameter to start up the partial nitrification process. After 86 days of operation, the nitrite accumulation rate reached 1.44 kg/(m day) in the SBR, and the removal efficiency of ammonia nitrogen (NH -N) was over 95 %. The PNGs showed a dense and compact structure, with an excellent settling ability, a typical extracellular polymeric substance (EPS) composition, and a high ammonia oxidation activity. The high-throughput pyrosequencing results indicated that the microbial community structure in the granules was significantly influenced by the C/N ratio, and ammonia-oxidizing bacteria (AOB), including the r-strategist Nitrosomonas and k-strategist Nitrosospira genre, which accounted for approximately 40 % of the total biomass at the end of operation. The effective suppression of nitrite-oxidizing bacteria (NOB) growth was attributed to oxygen competition on the granular surface among functional bacteria, as well as the high free ammonia or free nitrous acid concentrations during the aeration period. [ABSTRACT FROM AUTHOR]

When aerobic granular sludge is applied for industrial wastewater treatment, different soluble substrates can be present. For stable granular sludge formation on volatile fatty acids (e.g. acetate), production of storage polymers under anaerobic feeding conditions has been shown to be important. This prevents direct aerobic growth on readily available chemical oxygen demand (COD), which is thought to result in unstable granule formation. Here, we investigate the impact of acetate, methanol, butanol, propanol, propionaldehyde, and valeraldehyde on granular sludge formation at 35 °C. Methanogenic archaea, growing on methanol, were present in the aerobic granular sludge system. Methanol was completely converted to methane and carbon dioxide by the methanogenic archaeum Methanomethylovorans uponensis during the 1-h anaerobic feeding period, despite the relative high dissolved oxygen concentration (3.5 mg O L) during the subsequent 2-h aeration period. Propionaldehyde and valeraldehyde were fully disproportionated anaerobically into their corresponding carboxylic acids and alcohols. The organic acids produced were converted to storage polymers, while the alcohols (produced and from influent) were absorbed onto the granular sludge matrix and converted aerobically. Our observations show that easy biodegradable substrates not converted anaerobically into storage polymers could lead to unstable granular sludge formation. However, when the easy biodegradable COD is absorbed in the granules and/or when the substrate is converted by relatively slow growing bacteria in the aerobic period, stable granulation can occur. [ABSTRACT FROM AUTHOR]

A process involving the use of membrane bioreactor seeded with aerobic granular sludge (GMBR) was applied to the treatment of sewage containing pharmaceuticals and personal care products (PPCPs). The removal effects of five kinds of medicines in the reactor were investigated, and the microbial communities were constructed by polymerase chain reaction and denaturing gradient gel electrophoresis. We also determined the effects of different sludge retention and hydraulic retention times (SRT and HRT, respectively) and influent organic loading on GMBR's efficiency in processing sewage containing PPCPs. The removal effects of the GMBR on five PPCPs varied. Using the GMBR, the removal rates of prednisolone, naproxen and norfloxacin were 98.56, 84.02 and 87.85 %, respectively. The removal rates of sulfamethoxazole and ibuprofen were 77.83 and 63.32 %, respectively. In the system, PPCP drugs had relatively less effect on microbial diversity. A certain succession was observed in the structural variation of microbial species in the GMBR. Microorganisms that can degrade PPCPs gradually accumulated, and antibiotic-resistant microorganisms, such as Firmicutes sp., Aeromonas sp. and Nitrospira sp., served a key function in the treatment of sewage containing antibiotics. Long SRT and HRT during the GMBR process can facilitate the removal of most PPCPs. The system efficiently removed PPCPs at high influent organic loading. [ABSTRACT FROM AUTHOR]

Salinity can adversely affect the performance of most biological processes involved in wastewater treatment. The effect of salt on the main conversion processes in an aerobic granular sludge (AGS) process accomplishing simultaneous organic matter, nitrogen, and phosphate removal was evaluated in this work. Hereto, an AGS sequencing batch reactor was subjected to different salt concentrations (0.2 to 20 g Cl l). Granular structure was stable throughout the whole experimental period, although granule size decreased and a significant effluent turbidity was observed at the highest salinity tested. A weaker gel structure at higher salt concentrations was hypothesised to be the cause of such turbidity. Ammonium oxidation was not affected at any of the salt concentrations applied. However, nitrite oxidation was severely affected, especially at 20 g Cl l, in which a complete inhibition was observed. Consequently, high nitrite accumulation occurred. Phosphate removal was also found to be inhibited at the highest salt concentration tested. Complementary experiments have shown that a cascade inhibition effect took place: first, the deterioration of nitrite oxidation resulted in high nitrite concentrations and this in turn resulted in a detrimental effect to polyphosphate-accumulating organisms. By preventing the occurrence of the nitrification process and therefore avoiding the nitrite accumulation, the effect of salt concentrations on the bio-P removal process was shown to be negligible up to 13 g Cl l. Salt concentrations equal to 20 g Cl l or higher in absence of nitrite also significantly reduced phosphate removal efficiency in the system. [ABSTRACT FROM AUTHOR]

The aim of this study was to evaluate the impact of zeolite powders on feasibility of rapid aerobic granulation in the column-type sequencing batch reactors. After 90 days' operation, aerobic granular sludge was formed in both reactors by altering influent chemical oxygen demand/nitrogen (COD/N) ratios. R1 with zeolite powders had better removal capabilities of COD and total nitrogen than R2, which was without zeolite powders. Mixed liquor volatile suspended solid concentrations of the two reactors were 7.36 and 5.45 g/L, while sludge volume index (SVI) values were 34.9 and 47.9 mg/L, respectively. The mean diameters of aerobic granular sludge in the above two reactors were 2.5 and 1.5 mm, respectively. Both reactors achieved the largest simultaneous nitrification and denitrification (SND) efficiency at an influent COD/N ratio of 8; however, R1 exhibited more excellent SND efficiency than R2. The obtained results could provide a novel technique for rapid aerobic granulation and N removal simultaneously, especially when treating nitrogen-rich industrial wastewater. [ABSTRACT FROM AUTHOR]

In the present study, the factors influencing density of granular sludge particles were evaluated. Granules consist of microbes, precipitates and of extracellular polymeric substance. The volume fractions of the bacterial layers were experimentally estimated by fluorescent in situ hybridisation staining. The volume fraction occupied by precipitates was determined by computed tomography scanning. PHREEQC was used to estimate potential formation of precipitates to determine a density of the inorganic fraction. Densities of bacteria were investigated by Percoll density centrifugation. The volume fractions were then coupled with the corresponding densities and the total density of a granule was calculated. The sensitivity of the density of the entire granule on the corresponding settling velocity was evaluated by changing the volume fractions of precipitates or bacteria in a settling model. Results from granules originating from a Nereda reactor for simultaneous phosphate COD and nitrogen removal revealed that phosphate-accumulating organisms (PAOs) had a higher density than glycogen-accumulating organisms leading to significantly higher settling velocities for PAO-dominated granules explaining earlier observations of the segregation of the granular sludge bed inside reactors. The model showed that a small increase in the volume fraction of precipitates (1-5 %) strongly increased the granular density and thereby the settling velocity. For nitritation-anammox granular sludge, mainly granular diameter and not density differences are causing a segregation of the biomass in the bed. [ABSTRACT FROM AUTHOR]

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. [ABSTRACT FROM AUTHOR]

Aerobic granular sludge was successfully cultivated with the effluent of internal circulation reactor in a pilot-scale sequencing batch reactor (SBR). Soy protein wastewater was used as an external carbon source for altering the influent chemical oxygen demand/nitrogen (COD/N) ratios of SBR. Initially, the phenomenon of partial nitrification was observed and depressed by increasing the influent COD/N ratios from 3.32 to 7.24 mg/mg. After 90 days of aerobic granulation, the mixed liquor suspended solids concentration of the reactor increased from 2.80 to 7.02 g/L, while the sludge volumetric index decreased from 105.51 to 42.99 mL/g. The diameters of mature aerobic granules vary in the range of 1.2 to 2.0 mm. The reactor showed excellent removal performances for COD and $$ {\text{NH}}_4^{ + }{\text{ - N}} $$ after aerobic granulation, and average removal efficiencies were over 93% and 98%, respectively. The result of this study could provide further information on the development of aerobic granule-based system for full-scale applications. [ABSTRACT FROM AUTHOR]

In this study, we analysed the nitrifying microbial community (ammonium-oxidizing bacteria (AOB) and nitrite-oxidizing bacteria (NOB)) within three different aerobic granular sludge treatment systems as well as within one flocculent sludge system. Granular samples were taken from one pilot plant run on municipal wastewater as well as from two lab-scale reactors. Fluorescent in situ hybridization (FISH) and quantitative PCR (qPCR) showed that Nitrobacter was the dominant NOB in acetate-fed aerobic granules. In the conventional system, both Nitrospira and Nitrobacter were present in similar amounts. Remarkably, the NOB/AOB ratio in aerobic granular sludge was elevated but not in the conventional treatment plant suggesting that the growth of Nitrobacter within aerobic granular sludge, in particular, was partly uncoupled from the lithotrophic nitrite supply from AOB. This was supported by activity measurements which showed an approximately threefold higher nitrite oxidizing capacity than ammonium oxidizing capacity. Based on these findings, two hypotheses were considered: either Nitrobacter grew mixotrophically by acetate-dependent dissimilatory nitrate reduction ( ping-pong effect) or a nitrite oxidation/nitrate reduction loop ( nitrite loop) occurred in which denitrifiers reduced nitrate to nitrite supplying additional nitrite for the NOB apart from the AOB. [ABSTRACT FROM AUTHOR]

The process of nitrification-denitrification via nitrite for nitrogen removal under real-time control mode was tested in two laboratory-scale sequencing batch reactors (SBRs) with flocculent activated sludge (R1) and aerobic granular sludge (R2) to compare operational performance and real-time control strategies. The results showed that the average ammonia nitrogen, total inorganic nitrogen (TIN), and chemical oxygen demand (COD) removal during aeration phase were 97.6%, 57.0%, and 90.1% in R2 compared with 98.6%, 48.7%, and 88.1% in R1. The TIN removed in both SBRs was partially due to the presence of simultaneous nitrification-denitrification via nitrite, especially in R2. The specific nitrification and denitrification rates in R2 were 0.0416 mgNH-N/gSS-min and 0.1889 mgNO-N/gSS-min, which were 1.48 times and 1.35 times that of R1. The higher rates for COD removal, nitrification, and denitrification were achieved in R2 than R1 with similar influent quality. Dissolved oxygen (DO), pH, and oxidization reduction potential, corresponding to nutrient variations, were used as diagnostic parameters to control the organic carbon degradation and nitrification-denitrification via nitrite processes in both SBRs. The online control strategy of granular SBR was similar to that of the SBR with flocculent activated sludge. However, a unique U-type pattern on the DO curve in granular SBR was different from SBR with flocculent activated sludge in aerobic phase. [ABSTRACT FROM AUTHOR]

Two bubble column sequencing batch reactors fed with an artificial wastewater were operated at 20 °C, 30 °C, and 35 °C. In a first stage, stable granules were obtained at 20 °C, whereas fluffy structures were observed at 30 °C. Molecular analysis revealed high abundance of the operational taxonomic unit 208 (OTU 208) affiliating with filamentous bacteria Leptothrix spp. at 30 °C, an OTU much less abundant at 20 °C. The granular sludge obtained at 20 °C was used for the second stage during which one reactor was maintained at 20 °C and the second operated at 30 °C and 35 °C after prior gradual increase of temperature. Aerobic granular sludge with similar physical properties developed in both reactors but it had different nutrient elimination performances and microbial communities. At 20 °C, acetate was consumed during anaerobic feeding, and biological phosphorous removal was observed when Rhodocyclaceae-affiliating OTU 214 was present. At 30 °C and 35 °C, acetate was mainly consumed during aeration and phosphorous removal was insignificant. OTU 214 was almost absent but the Gammaproteobacteria-affiliating OTU 239 was more abundant than at 20 °C. Aerobic granular sludge at all temperatures contained abundantly the OTUs 224 and 289 affiliating with Sphingomonadaceae indicating that this bacterial family played an important role in maintaining stable granular structures. [ABSTRACT FROM AUTHOR]

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. [ABSTRACT FROM AUTHOR]