Influence of different substrates on the formation and characteristics of aerobic granules in sequencing batch reactors

The effects of different substrates on the aerobic granulation process were studied using laboratory-scale sequencing batch reactors （SBRs）. Four parallel granules sequencing batch reactors （GSBR）： R1, R2, R3, and R4 were fed with acetate, glucose, peptone and fecula, respectively. Stable aerobic granules were successfully cultivated in R1, R2, R4, and smaller granules less than 500 μm were formed in R3. Morphology and the physic-chemical characteristics of aerobic granules fed with different carbon substrates were investigated by the four reactors operated under the same pressure. The aerobic granules in the four reactors were observed and found that peptone was the most stable one due to its good settleability even after a sludge age as short as 10 d. A strong correlation was testified between the characteristics of aerobic granules and the properties of carbon substrates. The stability of aerobic granules was affected by extracellular polymer substances （EPS） derived from microorganism growth during feast time fed with different carbon substrates, and the influence of the property of storage substance was greater than that of its quantity. Optimal carbon substrates, which are helpful in the cultivation and retention of well-settling granules and in the enhancement of the overall ability of the aerobic granules reactors, were found.

Cultivation of aerobic granules for simultaneous nitrification and denitrification by seeding different inoculated sludge

Cultivation of aerobic granules for simultaneous nitrification and denitrification in two sequencing batch airlift bioreactors was studied. Conventional activated floc and anaerobic granules served as main two inoculated sludge in the systems. Morphological variations of sludge in the reactors were observed. It was found that the cultivation of aerobic granules was closely associated with the kind of inoculated sludge. Round and regular aerobic granules were prevailed in both reactors, and the physical characteristics of the aerobic granules in terms of settling ability, specific gravity, and ratio of water containing were distinct when the inoculate sludge was different. Aerobic granules formed by seeding activated floc are more excellent in simultaneous nitrification and denitrification than that by aerobic granules formed from anaerobic granules. It was concluded that inoculated sludge plays a crucial role in the cultivation of aerobic granules for simultaneous nitrification and denitrification.

Characteristics of aerobic granules grown on glucose a sequential batch shaking reactor

Aerobic heterotrophic granular sludge was cultivated in a sequencing batch shaking reactor(SBSR) in which a synthetic wastewater containing glucose as carbon source was fed. The characteristics of the aerobic granules were investigated. Compared with the conventional activated sludge flocs, the aerobic granules exhibit excellent physical characteristics in terms of settleability, size, shape, biomass density, and physical strength. Scanning electron micrographs revealed that in mature granules little filamentous bacteria could be found, rod-shaped and coccoid bacteria were the dominant microorganisms.

Biodegradation of p-cresol by aerobic granules in sequencing batch reactor

The cultivation of aerobic granules in sequencing batch reactor for the biodegradation of p-cresol was studied. The reactor was started with 100 mg/L of p-cresol. Aerobic granules first appeared within one month of start up. The granules were large and strong and had a compact structure. The diameter of stable granules was in the range of 1-5 mm. The integrity coefficient and granules density was found to be 96% and 1046 kg/m3, raspectively, The settling velocity of granules was found to be in the range of 2×10^-2-6×10^-7 m/sec. The aerobic granules were able to degrade p-cresol upto 800 mg/L at a removal efficiency of 88%. Specific p-cresol degradation rate in aerobic granules followed Haldane model for substrate inhibition, High specific p-cresol degradation rate up to 0,96 g pcresol/（g VSS.day） were sustained upto p-cresol concentration of 400 mg/L, Higher removal efficiency, good settling characteristics of aerobic granules, makes sequencing batch reactor suitable for erLhaneing the microorganism potential for biodegradation of inhibitory compounds.

Performance and role of N-acyl-homoserine lactone(AHL)-based quorum sensing(QS) in aerobic granules

The present study investigated the relationship between N-acyl-homoserine lactone（AHL）-based quorum sensing（QS） and the physico-chemical properties of aerobic granules.Stable mature granules were observed in SBR2 and SBR3 with average diameters of 0.96,and1.49 mm,respectively. The sludge densities of aerobic granules in SBR2 and SBR3 were1.0246,and 1.0201 g/mL,respectively,which were higher than that of flocculent sludge in SBR1（1.0065 g/mL）. The results showed that the activity of AHL-based QS in SBR2 and SBR3 amounted to 2.4- and 2.1-fold induction,however,that in SBR1 with flocculent sludge was1.6-fold induction. In addition,the results also showed that the activity of AHL-based QS in the three reactors rose in the feast condition,and then dropped with the consumption of substrate. However,the activity of AHL-based QS in these three reactors recovered again in prolonged starvation. Furthermore,the results showed that the enhancement of AHL-based QS favored the extracellular polymeric substance production of microorganisms in activated sludge. Thus,it could be concluded that aerobic granules showed higher AHL-based QS than flocculent sludge,which resulted from the higher sludge density of aerobic granules than flocculent sludge. AHL-based QS was related to the metabolism energy in the feast condition; however,in prolonged starvation,microorganisms would emit more AHL-like molecules to protect themselves to resist starvation. Moreover,the enhancement of AHL-based QS favored the EPS component productivity of the microorganisms in activated sludge,which contributed to maintain the aerobic granular structure.

Approaching the binding between Cu（Ⅱ） and aerobic granules by a modified titration and p-XRF

Interactions between metals and activated sludge can substantially affect the fate and transport of heavy metals in wastewater treatment plants. Therefore, it is important to develop a simple, fast and efficient method to elucidate the interaction. In this study, a modified titration method with a dynamic mode was developed to investigate the binding of Cu（Ⅱ）, a typical heavy metal, onto aerobic granules. The titration results indicated that pH and ionic strength both had a positive effect on the biosorption capacity of the granular sludge. The/-XRF results demonstrated that the distribution of metals on the granular surface was heterogeneous, and Cu showed strong correlations and had the same ＂hot spots＂ positions with other metal ions （e.g., Ca, Mg, Fe etc.）. Ion exchange and complexing were the main mechanisms for the biosorption of Cu（Ⅱ） by aerobic granules. These results would be beneficial for better understanding of Cu（Ⅱ） migration and its fate in wastewater treatment plants.

Application of high OLR-fed aerobic granules for the treatment of low-strength wastewater: Performance, granule morphology and microbial community

Aerobic granules, pre-cultivated at the organic loading rate （OLR） of 3.0 kg COD/（m3 ·day）, were used to treat low-strength wastewater in two sequencing batch reactors at low OLRs of 1.2 and 0.6 kg COD/（m3 ·day）, respectively. Reactor performance, evolution of granule morphology, structure and microbial community at low OLRs under long-term operation （130 days） were investigated. Results showed that low OLRs did not cause significant damage to granule structure as a dominant granule morphology with size over 540 μm was maintained throughout the operation. Aerobic granules at sizes of about 750 μm were finally obtained at the low OLRs. The granule reactors operated at low OLRs demonstrated effective COD and ammonia removals （above 90%）, smaller granule sizes and less biomass. The contents of extracellular polymeric substances in the granules were decreased while the ratios of exopolysaccharide/exoprotein were increased （above 1.0）. The granules cultivated at the low OLRs showed a smoother surface and more compact structure than the seeded granules. A significant shift in microbial community was observed but the microbial diversity remained relatively stable. Confocal Laser Scanning Microscopy observation showed that the live cells were spread throughout the whole granule, while the dead cells were mainly concentrated in the outer layer of the granule, and the proteins, polysaccharides and lipids were mainly located in the central regime of the granule. In conclusion, granules cultivated at high OLRs show potential for treating low-strength organic wastewater steadily under long-term operation.

Influence of temperature on the characteristics of aerobic granulation in sequencing batch airlift reactors

Aerobic granular sludge was cultivated in sequencing batch airlift reactors （SBAR） at 25, 30, and 35℃, respectively. The effect of temperature on the granules characteristics was analyzed and the microbial community structures of the granules were probed using scanning electron microscope （SEM） and polymerase chain reaction-denaturing gradient gel electrophoresis （PCR-DGGE）. The results showed that 30℃ is optimum for matured granule cultivation, where the granules had a more compact structure, better settling ability and higher bioactivity, the oxygen utilization rate （OUR） reached 1.14 mg O2/（g MLVSS.min） with COD removal rate of 97% and TP removal rate of 75%. The removal efficiency of NH3-N increased from 68.5% to 87.5% along with the temperature increment from 25 to 35℃. The DGGE profiles revealed that the microbial community structure at 25℃ showed the least similarity with those at other temperatures. The sequencing results indicated that the majority of dominant microbes belonged to Actinobacteria and Proteobacterium. Thermomonas sp., Ottowia sp. and Curtobacteriurn ammoniigenes might play important roles at different temperatures, respectively.

treatment of swine wastewater in aerobic granular reactors： comparison of different seed granules as factors

The granulation process, physic-chemical properties, pollution removal ability and bacterial com- munities of aerobic granules with different feed-waste- water （synthetic wastewater, R1; swine wastewater, R2）, and the change trend of some parameters of two types of granules in long-term operated reactors treating swine wastewater were investigated in this experiment. The result indicated that aerobic granulation with the synthetic wastewater had a faster rate compared with swine waste- water and that full granulation in R 1 and R2 was reached on the 30th day and 39th day, respectively. However, although the feed wastewater also had an obvious effect on the biomass fraction and extracellular polymeric sub- stances of the aerobic granules during the granulation process, these properties remained at a similar level after long-term operation. Moreover, a similar increasing trend could also be observed in terms of the nitrogen removal efficiencies of the aerobic granules in both reactors, and the average specific removal rates of the organics and ammonia nitrogen at the steady-state stage were 35.33mg.g^-1 VSS and 51.46mg.g^-1 VSS for R1, and 35.47mg.g^-1 VSS and 51.72mg.g^-1 VSS for R2, respectively. In addition, a shift in the bacterial diversity occurred in the granulation process, whereas bacterial communities in the aerobic granular reactor were not affected by the seed granules after long-term operation.

Granulation of filamentous microorganisms in a sequencing batch reactor with saline wastewater

Proliferation of filamentous microorganisms frequently leads to operational failure for activate sludge systems. In this study, it was found that filamentous microorganisms could grow in compact granular structure with 5% sodium chloride in the substrate. In the early period of experiment, coccoid and rode-like bacteria predominated in the yellowish-brown granules, and later the white and the black granules were developed by filamentous microorganisms. The filamentous granules exhibited low porosity and fast settling velocity, and were more compact even than bacteria granules. It was hypothesized that the elevated pH in the later period might be a possible reason for the compact growth of filamentous granules. However, the bacteria granules showed the high bioactivity in terms of specific oxygen utilizing rate, and comprised of a wider diversity of compounds based on the thermogravimetric evaluation. The findings in this study demonstrated that filamentous microbes could form compact granular structure, which may encourage the utilization of filamentous microorganisms rather than the inhibition of their growth, as the latter is frequently used for sludge bulking control.

Influence of microbial community structure of seed sludge on the properties of aerobic nitrifying granules

In order to evaluate the influence of microbial community structure of seed sludge on the properties of aerobic nitrifying granules, these granules were cultivated with different seed sludge, and the variation of microbial community and dominant bacterial groups that impact the nitrogen removal efficiency of the aerobic nitrifying granules were analyzed and identified using 16 s rDNA sequence and denaturing gradient gel electrophoresis（DGGE） profiles. The results presented here demonstrated that the influence of the community structure of seed sludge on the properties of aerobic nitrifying granules was remarkable, and the granules cultivated by activated sludge from a beer wastewater treatment plant showed better performance, with a stable sludge volume index（SVI） value of 20 m L/g, high extracellular polymeric substance（EPS） content of 183.3 mg/L, high NH4＋-N removal rate of 89.42% and abundant microbial population with 10 dominant bacterial groups. This indicated that activated sludge with abundant communities is suitable for use as seed sludge in culturing aerobic nitrifying granules.

Effect of wastewater COD/N ratio on aerobic nitrifying sludge granulation and microbial population shift

The effect of COD/N ratio on the granulation process and microbial population succession was investigated.Four identical sequencing batch reactors,R1,R2,R3 and R4,were operated with various initial COD/N ratios ranging from 0/200 to 800/200（m/m）.Ethanol was fed as the source of COD.Aerobic granules were successfully cultivated in R2 and R3,operating with the COD/N ratio of 200/200 and 400/200,respectively.Scanning electron microscope observations indicated that short rod-shaped and spherical bacteria were dominant in R2,while granules produced in R3 were surrounded with a large amount of filamentous bacteria.The average specific nitritation rate in R2 and R3 were 0.019 and 0.008 mg N/（mg MLVSS.hr）,respectively.Fluorescence in situ hybridization results demonstrated that nitrifying bacteria population was enriched remarkably in R2.It indicated that nitrification ability and nitrifying bacteria population were enriched remarkably at low COD/N ratio.However,no granules were formed in R1 and R4 which might attribute to either limited or excessive extracellular polymeric substances production.This study contributed to a better understanding of the role of COD/N ratio in nitrifying sludge granulation.

Simultaneous pyridine biodegradation and nitrogen removal in an aerobic granular system

Simultaneous pyridine biodegradation and nitrogen removal were successfully achieved in a sequencing batch reactor（SBR） based on aerobic granules. In a typical SBR cycle, nitritation occurred obviously after the majority of pyridine was removed, while denitrification occurred at early stage of the cycle when oxygen consumption was aggravated. The effect of several key operation parameters, i.e., air flow rate, influent NH4~＋-N concentration,influent p H and pyridine concentration, on nitritation, pyridine degradation and total nitrogen（TN） removal, was systematically investigated. The results indicated that high air flow rate had a positive effect on both pyridine degradation and nitritation but a negative impact of overhigh air flow rate. With the increase of NH4~＋ dosage, both nitritation and TN removal could be severely inhibited. Slightly alkaline condition, i.e., pH 7.0–8.0, was beneficial for both pyridine degradation and nitritation. High pyridine dosage often resulted in the delay of both pyridine degradation and nitritation. Besides, extracellular polymeric substances production was affected by air flow rate, NH4~＋ dosage, pyridine dosage and p H.In addition, high-throughput sequencing analysis demonstrated that Bdellovibrio and Paracoccus were the dominant species in the aerobic granulation system. Coexistence of pyridine degrader, nitrification related species, denitrification related species, polymeric substances producer and self-aggregation related species was also confirmed by highthroughput sequencing.

Effects of activated sludge flocs and pellets seeds on aerobic granule properties

Aerobic granules seeded with activated sludge flocs and pellets （obtained from activated sludge flocs） were cultivated in two sequencing batch reactors and their characteristics were compared. Compared with granules seeded with activated sludge flocs, those seeded with pellets had shorter start-up time, larger diameter, better chemical oxygen demand removal efficiency, and higher hydrophobicity, suspended solid concentration, and Mg 2＋ content. The different inocula led the granule surface with different microbial morphologies, but did not result in different distribution patterns of extracellular polymeric substances and cells. The anaerobic bacterium Anoxybacillus sp. was detected in the granules seeded with pellets. These results highlighted the advantage of pellet over activated sludge floc as the seed for aerobic granulation and wastewater treatment.

Qualitatively and quantitatively assessing the aggregation ability of sludge during aerobic granulation process combined XDLVO theory with physicochemical properties

Inexact mechanism of aerobic granulation still impedes optimization and application of aerobic granules. In this study, the extended Derjaguin, Landau, Verwey, and Overbeek（XDLVO） theory and physicochemical properties were combined to assess the aggregation ability of sludge during aerobic granulation process qualitatively and quantitatively. Results show that relative hydrophobicity of sludge and polysaccharide content of extracellular polymeric substances（EPS） increased, while electronegativity of sludge decreased during acclimation phase. After 20 days＇ acclimation, small granules began to form due to high aggregation ability of sludge. Since then, coexisted flocs and granules possessed distinct physicochemical properties during granulation and maturation phase. The relative hydrophobicity decreased while electronegativity increased for flocs, whereas that for granules presented reverse trend. Through analyzing the interaction energy using the XDLVO theory, small granules tended to self-grow rather than self-aggregate or attach of flocs due to poor aggregation ability between flocs and granules during the granulation phase. Besides, remaining flocs were unlikely to self-aggregate owing to poor aggregation ability, low hydrophobicity and high electronegativity.

Rapid aerobic granulation in an SBR treating piggery wastewater by seeding sludge from a municipal WWTP

Aerobic sludge granulation was rapidly obtained in the erlenmeyer bottle and sequencing batch reactor（SBR） using piggery wastewater.Aerobic granulation occurred on day 3 and granules with mean diameter of 0.2 mm and SVI_（30） of 20.3 mL/g formed in SBR on day 18.High concentrations of Ca and Fe in the raw piggery wastewater and operating mode accelerated aerobic granulation,even though the seed sludge was from a municipal wastewater treatment plant（WWTP）.Alpha diversity analysis revealed Operational Taxonomic Units,Shannon,ACE and Chao 1 indexes in aerobic granules were 2013,5.51,4665.5 and 3734.5,which were obviously lower compared to seed sludge.The percentages of major microbial communities,such as Protect）acteria,Bacteroidetes and Firmicutes were obviously higher in aerobic granules than seed sludge.Chlorqflexi,Planctomycetes,Actinobactena,TM7 and Aridobacteria showed much higher abundances in the inoculum.The main reasons might be the characteristics of raw piggery wastewater and granule structure.

Alternating nitrogen feeding strategy induced aerobic granulation:Influencing conditions and mechanism

Effective cultivation of stable aerobic granular sludge(AGS)is a crucial step in the successful application of this technology,and the formation of AGS could be facilitated by some environmental stress conditions.Four identical sequencing batch reactors(SBRs)were established to investigate the aerobic granulation process under the same alternating ammonia nitrogen feeding strategy superimposed with different environmental conditions(inorganic carbon source,temperature,N/COD).Although various superimposed conditions induced a significant difference in the size,settling velocity,mechanic strength of AGS,mature aerobic granules could be successfully obtained in all four reactors after 70 days’operation,indicating the alternating ammonia nitrogen feeding strategy was the most critical factor for AGS formation.Based on the results of redundancy analysis,the presence of an inorganic carbon source could facilitate the cultivation of AGS with nitrification function,while the moderate temperature and fluctuant N/COD might benefit the cultivation of more stable AGS.In addition,superimposed stress conditions could result in the difference in the microbial population between four reactors,but the population diversity and abundance of microorganisms were not the determinants of AGS formation.This study provided an effective method for the cultivation of AGS by using alternating ammonia nitrogen feeding strategy.

Aerobic granulation of pure bacterial strain Bacillus thuringiensis

The objective of this study is to cultivate aerobic granules by pure bacterial strain,Bacillus thuringiensis,in a sequencing batch reactor.Stable granules sized 2.0–2.2 mm were formed in the reactor after a five-week cultivation.These granules exhibited excellent settling attributes,and degraded phenol at rates of 1.49 and 1.19 g phenol/(g VSS·d)at 250 and 1500 mg/L of phenol concentration,respectively.Confocal laser scanning microscopic test results show that Bacillus thuringiensis was distributed over the initial small aggregates,and the outer edge of the granule was away from the core regime in the following stage.