Size-effect on the physical characteristics of the aerobic granule in a SBR

Owing to a fast growth rate, aerobic granules display a wide range of sizes, approximately 0.3-5.0 mm in diameter. As the diameter increases, the aerobic granule undergoes serial morphological and physical changes that could cause problems to the reactor operation, a phenomenon which, however, has not been fully studied hitherto. In this study, aerobic granules from a sequencing batch reactor were mechanically separated into various size-categories in order to investigate their physical properties, including sludge volumetric index (SVI), settling velocity (sv), specific surface hydrophobicity, granule strength, total solids, percentage volatile solids and other structural properties. Also, the live and dead biomass distribution was examined under a confocal laser scanning microscope after treatment with nucleic acid viability stains. Regardless of size, the biomass (both live and dead) was densest in the outer layer of the granule, which was about 600+/-50 microm thick. The live cells appeared only in the peripheral zone, while dead biomass spread into the inner zone. The biomass distribution pattern justified the changing physical properties of the granules as they grew bigger. As size increased, the sv, granule total density and biomass density increased but not in parallel with the size increment, while the granule strength, specific surface hydrophobicity and SVI decreased. Nonetheless, beyond a threshold size (4.0 mm diameter), the granules presented peculiar values in those properties, deviating from the initial trends. This was due to both inner and outer structural changes. The physical properties associated significantly with the size factor, for which the correlation coefficients were above 0.67. In view of biological viability and physical properties, the operational size-range suggested for optimal performance and economically effective aerobic SBR granular sludge is a diameter of 1.0-3.0 mm.     

Sedimentological evolution in an UASB treating SYNTHES, a new representative synthetic sewage, at low loading rates

The changes in the sedimentological attributes of the sludge bed in an upflow anaerobic sludge blanket (UASB) reactor fed with a low-strength wastewater mimicking raw domestic sewage were assessed in this study. The reactor was inoculated with 250 ml of granular sludge from a full-scale UASB reactor. The organic loading rate (OLR) varied from 1 to 2 g COD/ld. During the half-year long study, the reactor was operated at hydraulic retention times (HRTs) of 4.8 and 10 h, at 33 degrees C. Sludge sedimentology showed that the original granular sludge experienced serious instability and disintegration, leading to a much finer final grain assemblage, mainly due to substrate transfer limitation and cell starvation at the interior of larger granules. With time, the size uniformity tended to decrease, sphericity tended to increase, the skewness of the granule size distribution became negative, and the kurtosis became peaked and leptokurtic. In spite of the observed size reduction, reactor efficiency increased to a CODtotal removal of 96%. Biomass (sludge) yield was 0.012 g VS/g COD removed. The CH4 content of the biogas was high (up to 96%). This study thus highlights the treatment of a new type of wastewater with the deployment of the UASB reactor. It also reports the evolutionary trend of the biomass particle size distribution, making reference to a classic sedimentological appraisal.     

Different types of sludge granules in UASB reactors treating acidified wastewaters

The influence of a high energy substrate, i.e. sucrose, on the granular sludge yield and the development of different types of granular sludge was investigated by using Upflow Anaerobic Sludge Bed (UASB) reactors fed with synthetic wastewater. The feed COD was a mixture of volatile fatty acids (VFA) i.e., 20, 40, and 40% of the COD as C₂-, C₃-, and C₄-VFA, respectively. Furthermore, experiments were carried out in which 10 and 30% of the VFA COD was substituted with sucrose. The following distinctly different types of granules were observed in each testrun: in the reactor fed with solely VFA, black (B) and white (W) granules developed; in the reactor fed with a mixture of 90% VFA and 10% sucrose, three types of granules i.e., B, W, and grey (G) granules could be seen; in the reactor fed with 70% VFA and 30% sucrose, only W and G granules were found. The granular sludge yield increased proportional to the amount of sucrose COD. At steady-state performance of the reactors, specific acidogenic (SAA) and methanogenic (SMA) activity tests on these granules revealed that B granules had the highest SMA with low SAA. The W granules had very high SMA with low SAA. G granules gave the highest SAA with a considerable SMA. Measurement of coenzyme F₄₂₀ revealed that B granules consist mainly of acetoclastic methanogens. The fore-mentioned tests were supplemented with analyses of the wash-out cells present in the reactor effluent and the results suggested that acidogens, if present, prevail at the granule surface. The B granules were particularly rich in Ca, Mn, and Zn minerals. The size distribution analysis showed that the granule diameter increased in the following order: B相似文献   

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).     

Microbial community in granules from a high-rate EGSB reactor

The spatial distribution, quantity and diversity of different microorganisms within anaerobic granular sludge from a lab-scale expanded granular sludge bed (EGSB) reactor operated at different organic loading rates were studied using florescent in situ hybridization (FISH), real time quantitative — polymerase chain reaction (RTQ-PCR) and denaturing gradient gel electrophoresis (DGGE) techniques. The results indicated that most Eubacteria were located in the outer layer of granule, while the Archaea which mainly were methanogens and more sensible to the environmental conditions were located in the inner layer of the granule. The quantity of Archaea was obviously less than that of Eubacteria in the granules, but increased with the increasing of organic loading rates of the reactor. As the organic loading rate of the reactor increased and the operating time elapsed, the Archaea community in the granules changed significantly. Seven typical DGGE bands were collected and sequenced, and found that the dominant species of Archaea in the granules operated in the last period were mainly Methanocorpusculum, Methanobacterium, Methanosaeta.     

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.     

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.     

Aerobic granulation with brewery wastewater in a sequencing batch reactor

Aerobic granular sludge was cultivated in a sequencing batch reactor fed with brewery wastewater. After nine-week operation, stable granules with sizes of 2-7 mm were obtained. With the granulation, the SVI value decreased from 87.5 to 32 mL/g. The granular sludge had an excellent settling ability with the settling velocity over 91 m/h. Aerobic granular sludge exhibited good performance in the organics and nitrogen removal from brewery wastewater. After granulation, high and stable removal efficiencies of 88.7% COD(t), 88.9% NH(4)(+)-N were achieved at the volumetric exchange ratio of 50% and cycle duration of 6h. The average COD(t) and COD(s) of the effluent were 212 and 134 mg/L, respectively, and the average effluent ammonium concentration was less than 14.4 mg/L. Nitrogen was removed due to nitrification and simultaneous denitrification in the inner core of granules.     

Microbial population dynamics during sludge granulation in an anaerobic-aerobic biological phosphorus removal system

The evolution of a microbial community was investigated during sludge granulation using a wide range of micro-scale and molecular biology techniques. Experimental results demonstrate that polyphosphate-accumulating granules were successfully cultured during the anaerobic/aerobic cycle. Improvement in sludge sedimentation performance occurred prior to the formation of granular sludge and was not affected by change in granule size. Rod-shaped and filamentous bacteria appeared to initiate granule formation and generate the structures that supported further granule growth. It was observed that mature granules supported microbial populations that differed from nascent granules and were predominantly packed with coccoid bacteria. It was further observed that the diversity of the granular microbial community increased as the granules grew. Accumulibacter, Nitrosospira and Thauera were mainly responsible for nutrient removal while microorganisms such as Rhodocyclus and Hyphomicrobiaceae appeared to be primarily responsible for forming and maintaining the granule structure.     

Required minimum granule size in UASB reactor and characteristics variation with size

The objective of this study was to define minimum size of bioparticles that could be classified as granules, to offer all advantages of granular sludge. Based on the theory of sedimentation, the minimum diameter bioparticles, which should be considered as granules was found out for specific gravity of sludge ranging between 1.01 and 1.05. For example, for specific gravity of 1.035 the minimum diameter of granules required for better sludge retention was 0.34 mm. The diameter based on this theory was evaluated by carrying out settling column analysis of a granular sludge obtained from lab-scale UASB reactor and verified with microscopic observation. To find out the effect of granules size on the nature of biodegradability, specific methanogenic activity (SMA) was carried out. It was observed that SMA increased with size of bioparticles tested in the range of 0.27-3.03 mm. The change in VSS/SS ratio and specific gravity was observed with size of granules. Consideration of variation in specific gravity with size of granules increased the degree of validation of sedimentation theory for the calculation of granules diameter.     

Layered structure of bacterial aggregates produced in an upflow anaerobic sludge bed and filter reactor

The ultrastructure of bacterial granules that were maintained in an upflow anaerobic sludge bed and filter reactor was examined. The reactor was fed a sucrose medium, and it was operated at 35 degrees C. Scanning and transmission electron microscopy revealed that the granular aggregates were three-layered structures. The exterior layer of the granule contained a very heterogeneous population that included rods, cocci, and filaments of various sizes. The middle layer consisted of a slightly less heterogeneous population than the exterior layer. A more ordered arrangement, made up predominantly of bacterial rods, was evident in this second layer. The third layer formed the internal core of the granules. It consisted of large numbers of Methanothrix-like cells. Large cavities, indicative of vigorous gas production, were evident in the third layer. On the basis of these ultrastructural results, a model that presents a possible explanation of granule development is offered.     

Layered structure of bacterial aggregates produced in an upflow anaerobic sludge bed and filter reactor.

The ultrastructure of bacterial granules that were maintained in an upflow anaerobic sludge bed and filter reactor was examined. The reactor was fed a sucrose medium, and it was operated at 35 degrees C. Scanning and transmission electron microscopy revealed that the granular aggregates were three-layered structures. The exterior layer of the granule contained a very heterogeneous population that included rods, cocci, and filaments of various sizes. The middle layer consisted of a slightly less heterogeneous population than the exterior layer. A more ordered arrangement, made up predominantly of bacterial rods, was evident in this second layer. The third layer formed the internal core of the granules. It consisted of large numbers of Methanothrix-like cells. Large cavities, indicative of vigorous gas production, were evident in the third layer. On the basis of these ultrastructural results, a model that presents a possible explanation of granule development is offered.     

Effect of medium composition and sludge removal on the production, composition, and architecture of thermophilic (55 degrees C) acetate-utilizing granules from an upflow anaerobic sludge blanket reactor.

A thermophilic upflow anaerobic sludge blanket (UASB) reactor degrading acetate was started by applying published methods (W. M. Wiegant and A. W. A. de Man, Biotechnol. Bioeng. 28:718-77, 1986) for production of granules dominated by Methanothrix spp. The reactor was inoculated with thermophilic digested sludge. No granules were observed during the first 7 months of start-up of the UASB reactor. However, after the concentrations of potassium, phosphate, ammonium, and magnesium in the medium were gradually increased, granules developed, indicating that there was a critical concentration of one or more of the ions required for production of granules from the starting material. After several years of stable operation, the effect of removing 60% of the granular sludge was investigated. Immunologic qualitative and quantitative studies showed that removal of the granular sludge resulted in an increase in the number of the predominant methanogens, antigenically related to Methanosarcina thermophila TM-1 and Methanosarcina mazeii S-6, and Methanobacterium thermoautotrophicum delta H and GC1. These changes were accompanied by modifications of the microanatomy of the granules, as demonstrated histochemically and immunohistochemically. The results indicated that different catabolic pathways dominated in different regions of the granules, i.e., acetate oxidation in the middle of the granules, where there is a low acetate concentration, and an aceticlastic reaction in the outer surfaces, with a high acetate concentration. The results also showed that removal of granules from a UASB reactor which has been under steady-state operation for a long period can improve the reactor's performance via formation of denser and larger granules with improved microbial activities.     

Modeling a granule‐based anaerobic ammonium oxidizing (ANAMMOX) process

A mathematical model was developed to describe the anaerobic ammonium oxidation (ANAMMOX) process in a granular upflow anaerobic sludge blanket (UASB) reactor. ANAMMOX granules were cultivated in the UASB reactor by seeding aerobic granules. The granule‐based reactor had a great N‐loading resistant capacity. The model simulation results on the 1‐year reactor performance matched the experimental data well. The yield coefficient for the growth and the decay rate coefficient of the ANAMMOX granules were estimated to be 0.164 g COD g^?1 N and 0.00016 h^?1, respectively. With this model, the effects of process parameters on the reactor performance were evaluated. Results showed that the optimum granule diameter for the maximum N‐removal should be between 1.0 and 1.3 mm and that the optimum N loading rate should be 0.8 kg N m^?3 d^?1. In addition, the substrate micro‐profiles in the ANAMMOX granules were measured with a microelectrode to explore the diffusion dynamics within the granules, and the measured profiles matched the predicted results well. Biotechnol. Bioeng. 2009;103: 490–499. © 2009 Wiley Periodicals, Inc.     

Structural analysis of anaerobic granules in a phase separated reactor by electron microscopy

This paper discusses the microbial community structure of anaerobic granules and the effect of phase separation in anaerobic reactor on the characteristics of granules. Electron micrographs revealed that the core of anaerobic granular sludge consists predominantly of Methanosaeta-like cells, a key microorganism in granulation process. Granules in the methanogenic dominant zone of the reactor were stable and densely packed with smooth regular surface. On the other hand, granules subjected to acidogenic activities were less stable structures with broken parts and an irregular fissured surface. Anaerobic granules consisted of a vast diversity of species from the outer surface to the core of the granule and possessed a multi-layered structure. Viruses in the granules suggests the presence of bacteriophage in the granular biomass. These could be responsible for destroying cells and weakening the internal structure of granules, and thus possibly causing the breaking of granules. The observation of protozoa-like microorganism on the exterior zone of granular structure is believed to play an important role as bacterial predator and control the growth of bacterial cells. The images observed in this study shows that anaerobic granule harbour diverse number of microbial species, and act differently in acidogenic and methanogenic microbial zones.     

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.     

Formation of aerobic granules and their PHB production at various substrate and ammonium concentrations

Aerobic granular sludge rich in polyhydroxybutyrate (PHB) was cultivated in a sequencing batch reactor (SBR) by seeding anaerobic granular sludge. The PHB content in aerobic granules was investigated and the experimental results reveal that both influent chemical oxygen demand (COD) and ammonium concentrations had a significant effect on the morphological characteristics and the PHB production of the aerobic granular sludge. At a COD and ammonium concentration of 750 mg/L and 8.5mg/L, respectively, the PHB content of the granules reached 44%, but their poor settling ability, as evidenced by a high sludge volume index, was observed. This was attributed to the outgrowth of filamentous bacteria on the granule surface. However, an increase in the ammonium concentration resulted in an elevated sludge concentration and a decrease in the PHB content in the granules. In this case, the aerobic granular sludge with a regular and compact structure was formed. The results suggest that, through controlling the COD and ammonium concentrations in the influent, the PHB-rich aerobic granular sludge with good settling ability could be cultivated.     

Characterization and evaluation of aerobic granules in sequencing batch reactor

In order to investigate the aerobic granules cultured under alternating aerobic and anoxic conditions, a sequencing batch reactor (SBR) was operated without the presence of a carrier material. Nitrification and denitrification occurred alternately in the SBR operation, with an increased nitrification efficiency of up to 97% and a high chemical oxygen demand (COD) removal efficiency of up to 95%. It was observed that physical characteristics of granule play an important role in the performance of the SBR process. Light microscopy was used to observe the time dependent development of the granules in the SBR. Based on the microscopic observations, some floc-like sludges remained in the form of a mixture with granules for 30 days of operation. Even though various granule sizes had been formed in the reactor after 50 days, the granule sizes were primarily from 1 +/- 0.35 to 1.3 +/- 0.45 mm, rarely exceeding 2 mm. The granules were analyzed by a combination of microelectrodes and fluorescent in situ hybridization (FISH), which provides more detailed information on what happens inside the granules. Based on their results, ammonia oxidizing bacteria (AOB) existed primarily in the upper and middle layers of the granule. Assuming a first-order reaction for nitrification, most of the nitrification is likely to occur from the surface to 300 microm into the granular thickness.     

Granules characteristics in the vertical profile of a full-scale upflow anaerobic sludge blanket reactor treating poultry slaughterhouse wastewater

The performance and the granules characteristics of a 450 m(3) -UASB reactor operating for 1228 days, treating poultry slaughterhouse wastewater with an average COD reduction of 85% was examined. Granules were sampled in three different positions along the vertical central line of the reactor, revealing variations in the concentration of volatile total solids. Although the reactor had been in operation for an extended period of time, granule sizes of 0.5-1.5 mm appeared to predominate. The hollow core was well defined for granules with sizes ranging from 2 to 3 mm in all the sampling ports. The granules exhibited no layered microbial distribution and were packed with different morphotype cells intertwined randomly throughout the cross-section. Methanogenic Archaea predominated in the granules taken from every sampling port along the reactor. The results indicated that the characterization of the granules is a useful tool for the adoption of operational strategies toward optimization of UASB reactors.     

Granular sludge formation in upflow anaerobic sludge blanket (UASB) reactors

The state of the art for upflow anaerobic sludge blanket (UASB) reactors is discussed, focusing on the microbiology of immobilized anaerobic bacteria and the mechanism of granule formation. The development of granular sludge is the key factor for successful operation of the UASB reactors. Criteria for determining if granular sludge has developed in a UASB reactor is given based on the densities and diameters of the granular sludge. The shape and composition of granular sludge can vary significantly. Granules typically have a spherical form with a diameter from 0.14 to 5 mm. The inorganic mineral content varies from 10 to 90% of the dry weight of the granules, depending on the wastewater composition etc. The main components of the ash are calcium, potassium, and iron. The extracellular polymers in the granular sludge are important for the structure and maintenance of granules, while the inorganic composition seems to be of less importance. The extracellular polymer content varies between 0.6 and 20% of the volatile suspended solids and consists mainly of protein and polysaccharides. Both Methanosaeta spp. (formerly Methanothrix) and Methanosarcina spp. have been identified as important aceticlastic methanogens for the initial granulation and development of granular sludge. Immunological methods have been used to identify other methanogens in the granules. The results have showed that, besides the aceticlastic methanogens Methanosaeta spp. and Methanosarcina spp., hydrogen and formate utilizing bacteria are also present, e.g., Methanobacterium formicicum, Methanobacterium thermoautotrophicum, and Methanobrevibacter spp. Microcolonies of syntrophic bacteria are often observed in the granules, and the significant electron transfer in these microcolonies occurs through interspecies hydrogen transfer. The internal organization of the various groups of bacteria in the granules depends on the wastewater composition and the dominating metabolic pathways in the granules. Internal organization is observed in granules where such an arrangement is beneficial for an optimal degradation of the wastewater. A four-step model is given for the initial development of granular sludge. (c) 1996 John Wiley & Sons, Inc.