The anaerobic ammonia oxidation(Anammox) granular sludge bed reactor was started up successfully with nitrifying granular sludge. During the operation, the nitrifying granular sludge was gradually converted into Anamm...The anaerobic ammonia oxidation(Anammox) granular sludge bed reactor was started up successfully with nitrifying granular sludge. During the operation, the nitrifying granular sludge was gradually converted into Anammox granular sludge with good settling property and high conversion activity. The Anammox reactor worked well with the shortest HRT of 2 43 h. Under the condition that HRT w as 6 39 h and influent concentration of ammonia and nitrite was 10 mmol/L, the removal of ammonia and nitrite was 97 17% and 100 00%, respectively. Corresponding volumetric total nitrogen loading rate and volumetric total nitrogen conversion rate were 100 83 mmol/(L·d) and 98 95 mmol/(L·d). The performance of Anammox reactor was efficient and stable.展开更多
Sludge granulation is considered to be the most critical parameter governing successful operation of an upflow anaerobic sludge blanket and expanded granular sludge bed (EGSB) reactors. Pre-granulated seeding sludge...Sludge granulation is considered to be the most critical parameter governing successful operation of an upflow anaerobic sludge blanket and expanded granular sludge bed (EGSB) reactors. Pre-granulated seeding sludge could greatly reduce the required startup time. Two lab-scale and a pilot-scale EGSB reactors were operated to treat Shaoxing Wastewater Treatment Plant (SWWTP) containing wastewater from real engineering printing and dyeing with high pH and sulfate concentration. The microbiological structure and the particle size distribution in aerobic excess sludge, sanitary landfill sludge digested for one year, and the granular sludge of EGSB reactor after 400 d of operation were analyzed through scanning electron microscopy (SEM) and sieves. The lab-scale EGSB reactor seeded with anaerobic sludge after digestion for one year in landfill showed obviously better total chemical oxygen demand (TCOD) removal efficiency than one seeded with aerobic excess sludge after cation polyacrylamide flocculation-concentration and dehydration. The TCOD removed was 470.8 mg/L in pilot scale EGSB reactor at short hydraulic retention time of 15 h. SEM of sludge granules showed that the microbiological structure of the sludge from different sources showed some differences. SEM demonstrated that Methanobacterium sp. was present in the granules of pilot-scale EGSB and the granular sludge produced by landfill contained a mixture of anaerobic/anoxic organisms in abundance. The particle size distribution in EGSB demonstrated that using anaerobic granular sludge produced by sanitary landfill as the seeding granular sludge was feasible.展开更多
Aerobic granular sludge was cultivated adopting internal-circulate sequencing batch airlift reactor. The contradistinctive experiment about short-term membrane fouling between aerobic granular sludge system and activa...Aerobic granular sludge was cultivated adopting internal-circulate sequencing batch airlift reactor. The contradistinctive experiment about short-term membrane fouling between aerobic granular sludge system and activated sludge system were investigated. The membrane foulants was also characterized by Fourier transform infrared (FTIR) spectroscopy technique. The results showed that the aerobic granular sludge had excellent denitrification ability; the removal efficiency of TN could reach 90%. The aerobic granular sludge could alleviate membrane fouling effectively. The steady membrane flux of aerobic granular sludge was twice as much as that of activated sludge system. In addition, it was found that the aerobic granular sludge could result in severe membrane pore-blocking, however, the activated sludge could cause severe cake fouling. The major components of the foulants were identified as comprising of proteins and polysaccharide materials.展开更多
In this study, the authors have investigated the effects of various factors on both aerobic and anaerobic degradation of 4-t-octylphenol (4-t-OP) in granular sludge. In comparison, the aerobic degradation rate was m...In this study, the authors have investigated the effects of various factors on both aerobic and anaerobic degradation of 4-t-octylphenol (4-t-OP) in granular sludge. In comparison, the aerobic degradation rate was much higher than that of anaerobic degradation. The optimal pH values for 4-t-OP degradation in granular sludge were 9 and 7 under aerobic and anaerobic conditions, respectively. And the degradation rate decreased with an increase in the initial 4-t-OP concentration. Addition of yeast extract or homologous compounds such as phenol also enhanced the 4-t-OP degradation, especially under the aerobic condition. To investigate the bacterial community in this study, the denaturing gradient gel electrophoresis (DGGE) method was applied, based on the primers, for the 16S rDNA V3 region of bacteria, γ-proteobacteria and bacillus were identified as the major species of sludge.展开更多
A lab-scale sequencing batch reactor (SBR) was set-up and the aerobic granular sludge was successfully incubated using anaerobic granular sludge as seed sludge. Nitrogen was partially removed by simultaneous nitrifi...A lab-scale sequencing batch reactor (SBR) was set-up and the aerobic granular sludge was successfully incubated using anaerobic granular sludge as seed sludge. Nitrogen was partially removed by simultaneous nitrification and denitrification (SND) via nitrite with free ammonia (FA) of about 10 mg/L. The denaturing gradient gel electrophoresis (DGGE) method was used to investigate community structure of α-Proteobacteria, β-Proteobacteria, ammonia oxidizing bacteria (AOB), and Nitrospira populations during start-up. The population sizes of bacteria, AOB and Nitrospira were examined using real-time PCR method. The analysis of community structure and Shannon index showed that stable structure of AOB population was obtained at day 35, while the communities of α- Proteobacteria, β-Proteobacteria, and Nitrospira became stable after day 45. At stable stage, the average cell densities were 1.1× 10^12, 2.2×10^10 and 1.0×10^10 cells/L for bacteria, AOB and Nitrospira, respectively. The relationship between characteristics of nitrifying bacteria community and nitrogenous substrate utilization constant was discussed by calculating Pearson correlation. Certain correlation seemed to exist between population size, biodiversity, and degradation constant. And the influence of population size might be greater than that of biodiversity.展开更多
In some industrial plants, wastewater was intermittently or seasonally generated. There may be periods during which wastewater treatment facilities have to be set into an idle phase over several weeks. When wastewater...In some industrial plants, wastewater was intermittently or seasonally generated. There may be periods during which wastewater treatment facilities have to be set into an idle phase over several weeks. When wastewater was generated again, the activated sludge flocs may have disintegrated. In this experiment, re-activation characteristics of aerobic granular sludge starved for 2 months were investigated. Specific oxygen utilization rate(SOUR) was used as an indicator to evaluate the metabolic activity of the sludge. The results revealed that aerobic granular sludge could be stored up to two months without running the risk of losing the integrity of the granules and metabolic potentials. The apparent color of aerobic granules stored at room temperature gradually turned from brownish-yellowish to gray brown. They appeared brownish-yellowish again 2 weeks after re-activation. The velocity and strength of granules after 2-month idle period could be fully restored about 3 weeks after re-activation. Metabolic activity, however, dropped to 15 8 mg O_2/(g MLVSS·h), i.e. 74 % reduction after 2 months of storage. After restarting the reactor, it took 2 weeks that SOUR of up to 48 5 mg O_2 /(g MLVSS·h) was achieved. A stable effluent COD concentration of less than 150 mg/L was achieved during the re-activation process.展开更多
Based on the successful performance of a lab-scale upflow anaerobic sludge blanket (UASB) reactor with the capacity of simultaneous methanogenesis and denitrification (SMD), the specific phylogenetic groups and co...Based on the successful performance of a lab-scale upflow anaerobic sludge blanket (UASB) reactor with the capacity of simultaneous methanogenesis and denitrification (SMD), the specific phylogenetic groups and community structure of microbes in the SMD granule in the UASB reactor were investigated by the construction of the Eubacteria and Archaea 16S rDNA clone libraries, fragment length polymorphism, and sequence blast. Real time quantitative-polymerase chain reaction (RTQ-PCR) technique was used to quantify the contents of Eubacteria and Archaea in the SMD granule. The contents of some special predominant methanogens were also investigated. The results indicated that the Methanosaeta and Methanobacteria were the predominant methanogens in all Archaea in the SMD granule, with contents of 71.59% and 22.73% in all 88 random Archaea clones, respectively. The diversity of Eubacteria was much more complex than that of Archaea. The low GC positive gram bacteria and ε-Protebacteria were the main predominant Eubacteria species in SMD granule, their contents were 49.62% and 12.03% in all 133 random Eubacteria clones respectively. The results of RTQ-PCR indicated that the content of Archaea was less than Eubacteria, the Archaea content in total microorganisms in SMD granule was about 27.6%.展开更多
Objective To investigate the effect of chemical oxygen demand (COD) concentrations on the anaerobic ammonium oxidation (ANAMMOX). Methods An Expanded Granular Sludge Bed (EGSB) reactor was used to cultivate the ...Objective To investigate the effect of chemical oxygen demand (COD) concentrations on the anaerobic ammonium oxidation (ANAMMOX). Methods An Expanded Granular Sludge Bed (EGSB) reactor was used to cultivate the granular sludge and to perform the ANAMMOX reaction in the bench scale experiment. NH4^+-N and NO2^--N were measured by using colorimetric method. NO3^+-N was analyzed by using the UV spectrophotometric method. COD measurement was based on digestion with potassium dichromate in concentrated sulphuric acid. Results When the COD concentrations in the reactors were 0 mg/L, 200 mg/L, 350 mg/L, and 550 mg/L, respectively, the NH4^+-N removal efficiency was 12.5%, 14.2%, 14.3%, and 23.7%; the removal amount of NO2-N was almost the same; the nitrate removal efficiency was 16.8%, 94.5%, 86.6%, and 84.2% and TN removal efficiency was 16.3%, 50.7%, 46.9%, and 50.4%, moreover, the COD removal efficiency was 85%, 65.7%, and 60%; the COD removal rate was 27.42, 61.88, and 97.8 mg COD/(h·L). Conclusion COD concentrations have a significant influence on anaerobic ammonium oxidation by granular sludge.展开更多
Terylene artificial silk printing and dyeing wastewater(TPD wastewater), containing averaged 710 mg/L terephthalic acid(TA) as the main carbon source and the character pollutant, was subjected to expanded granular slu...Terylene artificial silk printing and dyeing wastewater(TPD wastewater), containing averaged 710 mg/L terephthalic acid(TA) as the main carbon source and the character pollutant, was subjected to expanded granular sludge bed(EGSB) process. The stability of the EGSB process was firstly conducted by laboratory experiment. TA ionization was the predominated factor influencing the acid-base balance of the system. High concentration of TA in wastewater resulted in sufficient buffering capacity to neutralize the volatile fatty acids(VFA) generated from substrate degradation and provided strong base for anaerobic system to resist the pH decrease below 6.5. VFA and UFA caused almost no inhibition on the anaerobic process and biogas production except that pH was below 6.35 and VFA was at its maximum value. Along with the granulating of the activated sludge, the efficiency of organic removal and production rate of biogas increased gradually and became more stable. After start-up, the efficiency of COD removal increased to 57%—64%, pH stabilized in a range of 7.99—8.04, and production rate of biogas was relatively high and stable. Sludge granulating, suitable influent of pH and loading were responsible for the EGSB stability. The variation of VFA concentration only resulted in neglectable rebound of pH, and the inhibition from VFA could be ignored in EGSB. The EGSB reactor was stable for TPD wastewater treatment.展开更多
To characterize the effects of pentachlorophenol (PCP) on the performance and microbial community of aerobic granular sludge in sequencing batch reactor (SBR), the web-based terminal restriction fragment length polymo...To characterize the effects of pentachlorophenol (PCP) on the performance and microbial community of aerobic granular sludge in sequencing batch reactor (SBR), the web-based terminal restriction fragment length polymorphism (T-RFLP) and real-time PCR (RT- PCR) techniques were used to explore the bacterial community structure. When PCP increased from 0 to 50 mg/L, the COD removal rate changed little, while the ammonia removal rate dropped from 100% to 64.9%. The results of molecular characterization showed t...展开更多
A microbial community structure of granules harvested from an anaerobic sludge blanket reactor treating phenolic wastewater was investigated using fluorescence in situ hybridization(FISH)and clone library construction...A microbial community structure of granules harvested from an anaerobic sludge blanket reactor treating phenolic wastewater was investigated using fluorescence in situ hybridization(FISH)and clone library construction.Clones of Syntrophorhabdaceae and Cryptanaerobacter were observed to be responsible for phenol degradation.For accurate taxonomic assignment of Cryptanaerobacter clones,phylogenetic analysis using nearly full-length 16S ribosomal RNA(rRNA)gene sequences was necessary.Three oligonucleotide probes were designed to detect the following three taxonomic groups:Syntrophorhabdaceae,Cryptanaerobacter,and Syntrophus.FISH analysis of thin sections of anaerobic granules showed a random distribution of bacteria and archaea.However,a well-defined distribution of Syntrophorhabdaceae,Cryptanaerobacter,and Syntrophus was observed.Cryptanaerobacter and Syntrophus were found on the outer layer of the granules and were closely associated with each other,while Syntrophorhabdaceae was located in the deeper part of the granules.Such specific distribution of the bacteria is most likely due to their metabolic association and affinity for the substrate.Phenol degradation in the granular sludge was observed to be carried out in the following way.First,Cryptanaerobacter converts phenol to benzoate,which is then degraded by Syntrophus into acetate.This syntrophic degradation of phenol occurs near the surface of the granule,where the phenol concen-tration is high.In the deeper part of the granule,where the phenol concentration is lower,Syntrophorhabdaceae degrades phenol into acetate.We observed that Syntrophorhabdaceae is less likely to produce benzoate as an intermediate to feed the neighboring organisms,which contradicts the theo-ries presented by previous studies.展开更多
[Objective]We aimed to discuss the optimal conditions of sludge granulation on the basis of IC reactor.[Method]By using the formulated glucose wastewater,we studied the rapid cultivation of granular sludge as well as ...[Objective]We aimed to discuss the optimal conditions of sludge granulation on the basis of IC reactor.[Method]By using the formulated glucose wastewater,we studied the rapid cultivation of granular sludge as well as its influencing factors,so as to discuss the optimal conditions of sludge granulation.[Result]Through the static culture outside of IC reactor and dynamic culture in IC reactor,granular sludge appeared within a training cycle,with particle size of 1.0-1.5 mm.In IC reactor,when COD concentration of influent water was 5 000 mg /L,rising velocity was 0.9 m /h,pH was around 7,and auxiliary materials (like flocculant and active carbon) and mature granular sludge were added to the actor during the training process,the granulation process of sludge was accelerated.[Conclusion]The research could provide references for the rapid start-up of IC reactor.展开更多
The screening and identification of attachment genes is important to exploring the formation mechanism of biofilms at the gene level.It is helpful to the development of key culture technologies for aerobic granular sl...The screening and identification of attachment genes is important to exploring the formation mechanism of biofilms at the gene level.It is helpful to the development of key culture technologies for aerobic granular sludge(AGS).In this study,genome-wide sequencing and gene editing were employed for the first time to investigate the effects and functions of attachment genes in AGS.With the help of whole-genome analysis,ten attachment genes were screened from thirteen genes,and the efficiency of gene screening was greatly improved.Then,two attachment genes were selected as examples to further confirm the gene functions by constructing gene-knockout recombinant mutants of Stenotrophomonas maltophilia;when the two attachment genes were knocked out,the attachment potential was reduced by 50.67%and 43.93%,respectively.The results provide a new theoretical principle and efficient method for the development of AGS from the perspective of attachment genes.展开更多
Aerobic granular sludge was cultivated by using different kinds of seed sludge in sequencing batch airlift reactor. The influence of seed sludge on physical and chemical properties of granular sludge was studied; the ...Aerobic granular sludge was cultivated by using different kinds of seed sludge in sequencing batch airlift reactor. The influence of seed sludge on physical and chemical properties of granular sludge was studied; the microbial community structure was probed by using scanning electron microscope and polymerase chain reaction-denaturing gradient gel electrophoresis (PCR-DGGE). The results showed that seed sludge played an important role on the formation of aerobic granules. Seed sludge taken from beer wastewater treatment plant (inoculum A) was more suitable for cultivating aerobic granules than that of sludge from municipal wastewater treatment plant (inoculurn B). Cultivated with inoculum A, large amount of mature granules formed after 35 days operation, its SVI reached 32.75 mL/g, and SOUR of granular sludge was beyond 1.10 mg/(g.min). By contrast, it needed 56 days obtaining mature granules using inoculum B. DGGE profiles indicated that the dominant microbial species in mature granules were 18 and 11 OTU when inoculum A and B were respectively employed as seed sludge. The sequencing results suggested that dominant species in mature granules cultivated by inoculum A were Paracoccus sp., Devosia hwasunensi, Pseudoxanthomonas sp., while the dominant species were Lactococcus raffinolactis and Pseudomonas sp. in granules developed from inoculum B.展开更多
Aniline-degrading microbes were cultivated and acclimated with the initial activated sludge collected from a chemical wastewater treatment plant. During the acclimation processes, aerobic granular sludge being able to...Aniline-degrading microbes were cultivated and acclimated with the initial activated sludge collected from a chemical wastewater treatment plant. During the acclimation processes, aerobic granular sludge being able to effectively degrade aniline was successfully formed, from which a preponderant bacterial strain was isolated and named as AN1. Effects of factors including pH, temperature, and second carbon/nitrogen source on the biodegradation of aniline were investigated. Results showed that the optimal conditions for the biodegradation of aniline by the strain AN1 were at pH 7.0 and 28–35°C. At the optimal pH and temperature, the biodegradation rate of aniline could reach as high as 17.8 mg/(L·hr) when the initial aniline concentration was 400 mg/L. Further studies revealed that the addition of 1 g/L glucose or ammonium chloride as a second carbon or nitrogen source could slightly enhance the biodegradation efficiency from 93.0% to 95.1%–98.5%. However, even more addition of glucose or ammonium could not further enhance the biodegradation process but delayed the biodegradation of aniline by the strain AN1. Based on morphological and physiological characteristics as well as the phylogenetic analysis of 26S rDNA sequences, the strain AN1 was identified as Candida tropicalis.展开更多
The effects of different chemical oxygen demand(COD)concentrations on the anammox granular sludge with Bamboo Charcoal(BC)addition were evaluated in UASB reactor.The results showed that the average total nitrogen(TN)r...The effects of different chemical oxygen demand(COD)concentrations on the anammox granular sludge with Bamboo Charcoal(BC)addition were evaluated in UASB reactor.The results showed that the average total nitrogen(TN)removal efficiency was reduced from 85.9%to 81.4%when COD concentration was increased from 50 to 150 mg/L.However,the TN removal efficiency of BC addition reactors was dramatically 3.1%-6.4%higher than that without BC under different COD concentrations.The average diameter of granular sludge was 0.13 mm higher than that without BC.The settling velocity was increased by elevated COD concentration,while the EPS and VSS/SS were increased with BC addition.The high-throughput Miseq sequencing analyses revealed that the bacterial diversity and richness were decreased under COD addition,and the Planctomycetes related to anammox bacteria were Candidatus Brocadia and Candidatus Kuenenia.The Metagenomic sequencing indicated that the abundance of denitrification related functional genes all increased with elevated COD,while the abundance of anammox related functional genes of decreased.The func-tional genes related to anammox was hydrazine synthase encoding genes(hzsA,hzsB and hzsB).The average relative abundance of hzs genes in the reactor with BC addition was higher than the control at COD concentrations of 50 mg/L and 150 mg/L.The functional genes of denitrification mediated by BC were higher than those without BC throughout the operation phase.It is interesting to note that BC addition greatly enriched the related func-tional genes of denitrification and anammox.展开更多
Aerobic granules were formed in a conven- tional, continuous flow, completely mixed activated sludge system (CMAS). The reactor was inoculated with seed sludge containing few filaments and fed with synthetic municip...Aerobic granules were formed in a conven- tional, continuous flow, completely mixed activated sludge system (CMAS). The reactor was inoculated with seed sludge containing few filaments and fed with synthetic municipal wastewater. The settling time of the sludge and the average dissolved oxygen (DO) of the reactor were 2 h and 4.2 mg. L 1, respectively. The reactor was agitated by a stirrer, with a speed of 250r·min^-1, to ensure good mixing . The granular sludge had good settleability, and the sludge volume index (SVI) was between 50 and 90 mL ·g ^-1. The laser particle analyzer showed the diameter of the granules to be between 0.18 and 1.25 mm. A scanning electron microscope (SEM) investigation revealed the predominance of sphere-like and rod-like bacteria, and only few filaments grew in the granules. The microbial community structure of the granules was also analyzed by polymerase chain reaction-denaturing gradient gel electro- phoresis (PCR-DGGE). Sequencing analysis indicated the dominant species were α, β, and γ-Proteobacteria, Bacteroidetes, and Firmicutes. The data from the study suggested that aerobic granules could form, if provided with sufficient number of filaments and high shear force. It was also observed that a high height-to-diameter ratio of the reactor and short settling time were not essential for the formation of aerobic granular sludge.展开更多
Two identical full-scale biogas-lift reactors treating brewery wastewater were inoculated with different types of sludge to compare their operational conditions, sludge characteristics, and kinetic models at a mesophi...Two identical full-scale biogas-lift reactors treating brewery wastewater were inoculated with different types of sludge to compare their operational conditions, sludge characteristics, and kinetic models at a mesophilic temperature. One reactor (R1) started up with anaerobic granular sludge in 12 weeks and obtained a continuously average organic loading rate (OLR) of 7.4 kg chemical oxygen demand (COD)/(m3.day), COD removal efficiency of 80%, and effluent COD of 450 mg/L. The other reactor (R2) started up with residual activated sludge in 30 weeks and granulation accomplished when the reactor reached an average OLR of 8.3 kg COD/(m^3·day), COD removal efficiency of 90%, and effluent COD of 240 mg/L. Differences in sludge characteristics,biogas compositions, and biogas- lift processes may be accounted for the superior efficiency of the treatment performance of R2 over R1. Grau second-order and modified StoverKincannon models based on influent and effluent concentrations as well as hydraulic retention time were successfully used to develop kinetic parameters of the experimental data with high correlation coefficients (R2 〉 0.95), which further showed that R2 had higher treatment performance than R1. These results demonstrated that residual activated sludge could be used effectively instead of anaerobic granular sludge despite the need for a longer time.展开更多
Shortcut nitrification-denitrification,anaerobic ammonium oxidation(ANAMMOX),and methanogenesis have been successfully coupled in an Expanded Granular Sludge Bed-Biological Aerated Filter(EGSB-BAF)integrated system.As...Shortcut nitrification-denitrification,anaerobic ammonium oxidation(ANAMMOX),and methanogenesis have been successfully coupled in an Expanded Granular Sludge Bed-Biological Aerated Filter(EGSB-BAF)integrated system.As fed different synthetic wastewater with chemical oxygen demand(COD)of 300-1200 mg·L^(-1)and NH_(4)^(+)-N of 30-120 mg·L^(-1)at the outer recycle ratio of 200%,the influence of influent on ANAMMOX in the integrated system was investigated in this paper.The experimental results showed that higher COD concentration caused an increase in denitrification and methanogenesis but a decrease in ANAMMOX;however,when an influent with the low concentration of COD was used,the opposite changes could be observed.Higher influent NH_(4)^(+)-N concentration favored ANAMMOX when the COD concentration of influent was fixed.Therefore,low COD=NH_(4)^(+)-N ratio would decrease competition for nitrite between ANAMMOX and denitrification,which was favorable for reducing the negative effect of organic COD on ANAMMOX.The good performance of the integrated system indicated that the bacterial community of denitrification,ANAMMOX,and methanogenesis could be dynamically maintained in the sludge of EGSB reactor for a certain range of influent.展开更多
This paper reports the effects of shear force on anoxic granular sludge in sequencing batch reactors (SBR). The study was carried out in two SBRs (SBR1 and SBR2) in which sodium acetate (200mg COD·L^-1) was...This paper reports the effects of shear force on anoxic granular sludge in sequencing batch reactors (SBR). The study was carried out in two SBRs (SBR1 and SBR2) in which sodium acetate (200mg COD·L^-1) was used as the sole substrate and sodium nitrate (40 mgNO3-N·L^-1) was employed as the electron acceptor. The preliminary objective of this study was to cultivate anoxic granules in the SBR in order to investigate the effects of shear force on the formation of anoxic granular sludge and to compare the properties of anoxic sludge in the SBR. This study reports new results for the values of average velocity gradient, a measure of the applied shear force, which was varied in the two SBRs (3.79 s^-1 and 9.76 s^-1 for SBR1 and SBR2 respectively). The important findings of this research highlight the dual effects of shear force on anoxic granules. A low shear force can produce large anoxic granules with high activity and poor settling ability, whereas higher shear forces produce smaller granules with better settling ability and lower activity. The results of this study show that the anoxic granulation is closely related to the strength of the shear force. For high shear force, this research demonstrated that: 1) granules with smaller diameters, high density and good settling ability were formed in the reactor, and 2) granular sludge formed faster than it did in the low shear force reactor (41days versus 76 days). Once a steady-state has been achieved, the nitrate and COD removal rates were found to be 98% and 80%, respectively. For low shear force, such as was applied in SBR1, this research demonstrated that: 1) the activity of anoxic granular sludge in low shear force was higher than that in high shear force, 2) higher amount of soluble microbial products (SMPs) were produced, and 3) large pores were observed inside the larger granules,which are beneficial for nitrogen gas diffusion. Electron microscopic examination of the anoxic granules in both reactors showed that the morphology of the granules was ellipsoidal with a clear outline. Coccus and rod-shaped bacteria were wrapped by filamentous bacteria on the surface of granule.展开更多
文摘The anaerobic ammonia oxidation(Anammox) granular sludge bed reactor was started up successfully with nitrifying granular sludge. During the operation, the nitrifying granular sludge was gradually converted into Anammox granular sludge with good settling property and high conversion activity. The Anammox reactor worked well with the shortest HRT of 2 43 h. Under the condition that HRT w as 6 39 h and influent concentration of ammonia and nitrite was 10 mmol/L, the removal of ammonia and nitrite was 97 17% and 100 00%, respectively. Corresponding volumetric total nitrogen loading rate and volumetric total nitrogen conversion rate were 100 83 mmol/(L·d) and 98 95 mmol/(L·d). The performance of Anammox reactor was efficient and stable.
基金Project supported by the Major Scientific Key Problem Program of Scientific Commission of Zhejiang Province,China(2004C13027).
文摘Sludge granulation is considered to be the most critical parameter governing successful operation of an upflow anaerobic sludge blanket and expanded granular sludge bed (EGSB) reactors. Pre-granulated seeding sludge could greatly reduce the required startup time. Two lab-scale and a pilot-scale EGSB reactors were operated to treat Shaoxing Wastewater Treatment Plant (SWWTP) containing wastewater from real engineering printing and dyeing with high pH and sulfate concentration. The microbiological structure and the particle size distribution in aerobic excess sludge, sanitary landfill sludge digested for one year, and the granular sludge of EGSB reactor after 400 d of operation were analyzed through scanning electron microscopy (SEM) and sieves. The lab-scale EGSB reactor seeded with anaerobic sludge after digestion for one year in landfill showed obviously better total chemical oxygen demand (TCOD) removal efficiency than one seeded with aerobic excess sludge after cation polyacrylamide flocculation-concentration and dehydration. The TCOD removed was 470.8 mg/L in pilot scale EGSB reactor at short hydraulic retention time of 15 h. SEM of sludge granules showed that the microbiological structure of the sludge from different sources showed some differences. SEM demonstrated that Methanobacterium sp. was present in the granules of pilot-scale EGSB and the granular sludge produced by landfill contained a mixture of anaerobic/anoxic organisms in abundance. The particle size distribution in EGSB demonstrated that using anaerobic granular sludge produced by sanitary landfill as the seeding granular sludge was feasible.
基金Project supported by the University Doctorial Research Foundation.Ministry of Education of China(No.20030141022).
文摘Aerobic granular sludge was cultivated adopting internal-circulate sequencing batch airlift reactor. The contradistinctive experiment about short-term membrane fouling between aerobic granular sludge system and activated sludge system were investigated. The membrane foulants was also characterized by Fourier transform infrared (FTIR) spectroscopy technique. The results showed that the aerobic granular sludge had excellent denitrification ability; the removal efficiency of TN could reach 90%. The aerobic granular sludge could alleviate membrane fouling effectively. The steady membrane flux of aerobic granular sludge was twice as much as that of activated sludge system. In addition, it was found that the aerobic granular sludge could result in severe membrane pore-blocking, however, the activated sludge could cause severe cake fouling. The major components of the foulants were identified as comprising of proteins and polysaccharide materials.
文摘In this study, the authors have investigated the effects of various factors on both aerobic and anaerobic degradation of 4-t-octylphenol (4-t-OP) in granular sludge. In comparison, the aerobic degradation rate was much higher than that of anaerobic degradation. The optimal pH values for 4-t-OP degradation in granular sludge were 9 and 7 under aerobic and anaerobic conditions, respectively. And the degradation rate decreased with an increase in the initial 4-t-OP concentration. Addition of yeast extract or homologous compounds such as phenol also enhanced the 4-t-OP degradation, especially under the aerobic condition. To investigate the bacterial community in this study, the denaturing gradient gel electrophoresis (DGGE) method was applied, based on the primers, for the 16S rDNA V3 region of bacteria, γ-proteobacteria and bacillus were identified as the major species of sludge.
基金Project supported by the New Century Excellent Talent Scholarship of China(No.NCET-05-0387)the Doctodal Unit Scholarship of China(No.20050247016).
文摘A lab-scale sequencing batch reactor (SBR) was set-up and the aerobic granular sludge was successfully incubated using anaerobic granular sludge as seed sludge. Nitrogen was partially removed by simultaneous nitrification and denitrification (SND) via nitrite with free ammonia (FA) of about 10 mg/L. The denaturing gradient gel electrophoresis (DGGE) method was used to investigate community structure of α-Proteobacteria, β-Proteobacteria, ammonia oxidizing bacteria (AOB), and Nitrospira populations during start-up. The population sizes of bacteria, AOB and Nitrospira were examined using real-time PCR method. The analysis of community structure and Shannon index showed that stable structure of AOB population was obtained at day 35, while the communities of α- Proteobacteria, β-Proteobacteria, and Nitrospira became stable after day 45. At stable stage, the average cell densities were 1.1× 10^12, 2.2×10^10 and 1.0×10^10 cells/L for bacteria, AOB and Nitrospira, respectively. The relationship between characteristics of nitrifying bacteria community and nitrogenous substrate utilization constant was discussed by calculating Pearson correlation. Certain correlation seemed to exist between population size, biodiversity, and degradation constant. And the influence of population size might be greater than that of biodiversity.
文摘In some industrial plants, wastewater was intermittently or seasonally generated. There may be periods during which wastewater treatment facilities have to be set into an idle phase over several weeks. When wastewater was generated again, the activated sludge flocs may have disintegrated. In this experiment, re-activation characteristics of aerobic granular sludge starved for 2 months were investigated. Specific oxygen utilization rate(SOUR) was used as an indicator to evaluate the metabolic activity of the sludge. The results revealed that aerobic granular sludge could be stored up to two months without running the risk of losing the integrity of the granules and metabolic potentials. The apparent color of aerobic granules stored at room temperature gradually turned from brownish-yellowish to gray brown. They appeared brownish-yellowish again 2 weeks after re-activation. The velocity and strength of granules after 2-month idle period could be fully restored about 3 weeks after re-activation. Metabolic activity, however, dropped to 15 8 mg O_2/(g MLVSS·h), i.e. 74 % reduction after 2 months of storage. After restarting the reactor, it took 2 weeks that SOUR of up to 48 5 mg O_2 /(g MLVSS·h) was achieved. A stable effluent COD concentration of less than 150 mg/L was achieved during the re-activation process.
文摘Based on the successful performance of a lab-scale upflow anaerobic sludge blanket (UASB) reactor with the capacity of simultaneous methanogenesis and denitrification (SMD), the specific phylogenetic groups and community structure of microbes in the SMD granule in the UASB reactor were investigated by the construction of the Eubacteria and Archaea 16S rDNA clone libraries, fragment length polymorphism, and sequence blast. Real time quantitative-polymerase chain reaction (RTQ-PCR) technique was used to quantify the contents of Eubacteria and Archaea in the SMD granule. The contents of some special predominant methanogens were also investigated. The results indicated that the Methanosaeta and Methanobacteria were the predominant methanogens in all Archaea in the SMD granule, with contents of 71.59% and 22.73% in all 88 random Archaea clones, respectively. The diversity of Eubacteria was much more complex than that of Archaea. The low GC positive gram bacteria and ε-Protebacteria were the main predominant Eubacteria species in SMD granule, their contents were 49.62% and 12.03% in all 133 random Eubacteria clones respectively. The results of RTQ-PCR indicated that the content of Archaea was less than Eubacteria, the Archaea content in total microorganisms in SMD granule was about 27.6%.
基金The work was supported by the National Natural Science Foundation of China (Grant No. 59978020).
文摘Objective To investigate the effect of chemical oxygen demand (COD) concentrations on the anaerobic ammonium oxidation (ANAMMOX). Methods An Expanded Granular Sludge Bed (EGSB) reactor was used to cultivate the granular sludge and to perform the ANAMMOX reaction in the bench scale experiment. NH4^+-N and NO2^--N were measured by using colorimetric method. NO3^+-N was analyzed by using the UV spectrophotometric method. COD measurement was based on digestion with potassium dichromate in concentrated sulphuric acid. Results When the COD concentrations in the reactors were 0 mg/L, 200 mg/L, 350 mg/L, and 550 mg/L, respectively, the NH4^+-N removal efficiency was 12.5%, 14.2%, 14.3%, and 23.7%; the removal amount of NO2-N was almost the same; the nitrate removal efficiency was 16.8%, 94.5%, 86.6%, and 84.2% and TN removal efficiency was 16.3%, 50.7%, 46.9%, and 50.4%, moreover, the COD removal efficiency was 85%, 65.7%, and 60%; the COD removal rate was 27.42, 61.88, and 97.8 mg COD/(h·L). Conclusion COD concentrations have a significant influence on anaerobic ammonium oxidation by granular sludge.
文摘Terylene artificial silk printing and dyeing wastewater(TPD wastewater), containing averaged 710 mg/L terephthalic acid(TA) as the main carbon source and the character pollutant, was subjected to expanded granular sludge bed(EGSB) process. The stability of the EGSB process was firstly conducted by laboratory experiment. TA ionization was the predominated factor influencing the acid-base balance of the system. High concentration of TA in wastewater resulted in sufficient buffering capacity to neutralize the volatile fatty acids(VFA) generated from substrate degradation and provided strong base for anaerobic system to resist the pH decrease below 6.5. VFA and UFA caused almost no inhibition on the anaerobic process and biogas production except that pH was below 6.35 and VFA was at its maximum value. Along with the granulating of the activated sludge, the efficiency of organic removal and production rate of biogas increased gradually and became more stable. After start-up, the efficiency of COD removal increased to 57%—64%, pH stabilized in a range of 7.99—8.04, and production rate of biogas was relatively high and stable. Sludge granulating, suitable influent of pH and loading were responsible for the EGSB stability. The variation of VFA concentration only resulted in neglectable rebound of pH, and the inhibition from VFA could be ignored in EGSB. The EGSB reactor was stable for TPD wastewater treatment.
基金the Science Foundation ofJiangsu Province, China (No. BK2005402)the Nation-al Natural Science Foundation of China (No. 30640018)
文摘To characterize the effects of pentachlorophenol (PCP) on the performance and microbial community of aerobic granular sludge in sequencing batch reactor (SBR), the web-based terminal restriction fragment length polymorphism (T-RFLP) and real-time PCR (RT- PCR) techniques were used to explore the bacterial community structure. When PCP increased from 0 to 50 mg/L, the COD removal rate changed little, while the ammonia removal rate dropped from 100% to 64.9%. The results of molecular characterization showed t...
基金supported by Grant-in-Aids for Scientific Research(B)(JP18H01564)from the Japan Society for the Promotion of Science.
文摘A microbial community structure of granules harvested from an anaerobic sludge blanket reactor treating phenolic wastewater was investigated using fluorescence in situ hybridization(FISH)and clone library construction.Clones of Syntrophorhabdaceae and Cryptanaerobacter were observed to be responsible for phenol degradation.For accurate taxonomic assignment of Cryptanaerobacter clones,phylogenetic analysis using nearly full-length 16S ribosomal RNA(rRNA)gene sequences was necessary.Three oligonucleotide probes were designed to detect the following three taxonomic groups:Syntrophorhabdaceae,Cryptanaerobacter,and Syntrophus.FISH analysis of thin sections of anaerobic granules showed a random distribution of bacteria and archaea.However,a well-defined distribution of Syntrophorhabdaceae,Cryptanaerobacter,and Syntrophus was observed.Cryptanaerobacter and Syntrophus were found on the outer layer of the granules and were closely associated with each other,while Syntrophorhabdaceae was located in the deeper part of the granules.Such specific distribution of the bacteria is most likely due to their metabolic association and affinity for the substrate.Phenol degradation in the granular sludge was observed to be carried out in the following way.First,Cryptanaerobacter converts phenol to benzoate,which is then degraded by Syntrophus into acetate.This syntrophic degradation of phenol occurs near the surface of the granule,where the phenol concen-tration is high.In the deeper part of the granule,where the phenol concentration is lower,Syntrophorhabdaceae degrades phenol into acetate.We observed that Syntrophorhabdaceae is less likely to produce benzoate as an intermediate to feed the neighboring organisms,which contradicts the theo-ries presented by previous studies.
基金Supported by University Students' Innovation Project of Jiangsu Province,China (166201100022 )Scientific Research Foundation of Nanjing Institute of Technology (QKJC2010011)Science and Technology Support (Industrial) Project of Jiangsu Province,China(BE2010203)
文摘[Objective]We aimed to discuss the optimal conditions of sludge granulation on the basis of IC reactor.[Method]By using the formulated glucose wastewater,we studied the rapid cultivation of granular sludge as well as its influencing factors,so as to discuss the optimal conditions of sludge granulation.[Result]Through the static culture outside of IC reactor and dynamic culture in IC reactor,granular sludge appeared within a training cycle,with particle size of 1.0-1.5 mm.In IC reactor,when COD concentration of influent water was 5 000 mg /L,rising velocity was 0.9 m /h,pH was around 7,and auxiliary materials (like flocculant and active carbon) and mature granular sludge were added to the actor during the training process,the granulation process of sludge was accelerated.[Conclusion]The research could provide references for the rapid start-up of IC reactor.
基金supported by the National Natural Science Foundation of China(No.51578069)Beijing Science and Technology Commission Project(No.Z171100000717012)。
文摘The screening and identification of attachment genes is important to exploring the formation mechanism of biofilms at the gene level.It is helpful to the development of key culture technologies for aerobic granular sludge(AGS).In this study,genome-wide sequencing and gene editing were employed for the first time to investigate the effects and functions of attachment genes in AGS.With the help of whole-genome analysis,ten attachment genes were screened from thirteen genes,and the efficiency of gene screening was greatly improved.Then,two attachment genes were selected as examples to further confirm the gene functions by constructing gene-knockout recombinant mutants of Stenotrophomonas maltophilia;when the two attachment genes were knocked out,the attachment potential was reduced by 50.67%and 43.93%,respectively.The results provide a new theoretical principle and efficient method for the development of AGS from the perspective of attachment genes.
基金supported by the National Science Foundation of Heilongjiang Province (No. E200824)the Hi-Tech Research and Development Program (863) of China (No. 2002AA601310)
文摘Aerobic granular sludge was cultivated by using different kinds of seed sludge in sequencing batch airlift reactor. The influence of seed sludge on physical and chemical properties of granular sludge was studied; the microbial community structure was probed by using scanning electron microscope and polymerase chain reaction-denaturing gradient gel electrophoresis (PCR-DGGE). The results showed that seed sludge played an important role on the formation of aerobic granules. Seed sludge taken from beer wastewater treatment plant (inoculum A) was more suitable for cultivating aerobic granules than that of sludge from municipal wastewater treatment plant (inoculurn B). Cultivated with inoculum A, large amount of mature granules formed after 35 days operation, its SVI reached 32.75 mL/g, and SOUR of granular sludge was beyond 1.10 mg/(g.min). By contrast, it needed 56 days obtaining mature granules using inoculum B. DGGE profiles indicated that the dominant microbial species in mature granules were 18 and 11 OTU when inoculum A and B were respectively employed as seed sludge. The sequencing results suggested that dominant species in mature granules cultivated by inoculum A were Paracoccus sp., Devosia hwasunensi, Pseudoxanthomonas sp., while the dominant species were Lactococcus raffinolactis and Pseudomonas sp. in granules developed from inoculum B.
基金supported by the National Natural Science Foundation of China (No. 20977048)the National High Technology and Development Program (863) of China (No. 2009AA06Z317)a Project Funded by the Priority Academic Program Development of Jiangsu Higher Education Institutions (PAPD)
文摘Aniline-degrading microbes were cultivated and acclimated with the initial activated sludge collected from a chemical wastewater treatment plant. During the acclimation processes, aerobic granular sludge being able to effectively degrade aniline was successfully formed, from which a preponderant bacterial strain was isolated and named as AN1. Effects of factors including pH, temperature, and second carbon/nitrogen source on the biodegradation of aniline were investigated. Results showed that the optimal conditions for the biodegradation of aniline by the strain AN1 were at pH 7.0 and 28–35°C. At the optimal pH and temperature, the biodegradation rate of aniline could reach as high as 17.8 mg/(L·hr) when the initial aniline concentration was 400 mg/L. Further studies revealed that the addition of 1 g/L glucose or ammonium chloride as a second carbon or nitrogen source could slightly enhance the biodegradation efficiency from 93.0% to 95.1%–98.5%. However, even more addition of glucose or ammonium could not further enhance the biodegradation process but delayed the biodegradation of aniline by the strain AN1. Based on morphological and physiological characteristics as well as the phylogenetic analysis of 26S rDNA sequences, the strain AN1 was identified as Candida tropicalis.
基金supported by the China Postdoctoral Science Foundation (No. 2020M671400)the Natural Science Foundation of Jiangsu Province (No. BK20201450)+3 种基金the Jiangsu Qing Lan ProjectSuzhou Science and Technology Planning Project (Nos.SS202016 and SS2019022)the Opening Fund of Jiangsu Provincial Key Laboratory of Environmental Science and Engineering(No. Zd1804)the Graduate Research Innovation Program of Jiangsu Province (No. KYCX19_2030)
文摘The effects of different chemical oxygen demand(COD)concentrations on the anammox granular sludge with Bamboo Charcoal(BC)addition were evaluated in UASB reactor.The results showed that the average total nitrogen(TN)removal efficiency was reduced from 85.9%to 81.4%when COD concentration was increased from 50 to 150 mg/L.However,the TN removal efficiency of BC addition reactors was dramatically 3.1%-6.4%higher than that without BC under different COD concentrations.The average diameter of granular sludge was 0.13 mm higher than that without BC.The settling velocity was increased by elevated COD concentration,while the EPS and VSS/SS were increased with BC addition.The high-throughput Miseq sequencing analyses revealed that the bacterial diversity and richness were decreased under COD addition,and the Planctomycetes related to anammox bacteria were Candidatus Brocadia and Candidatus Kuenenia.The Metagenomic sequencing indicated that the abundance of denitrification related functional genes all increased with elevated COD,while the abundance of anammox related functional genes of decreased.The func-tional genes related to anammox was hydrazine synthase encoding genes(hzsA,hzsB and hzsB).The average relative abundance of hzs genes in the reactor with BC addition was higher than the control at COD concentrations of 50 mg/L and 150 mg/L.The functional genes of denitrification mediated by BC were higher than those without BC throughout the operation phase.It is interesting to note that BC addition greatly enriched the related func-tional genes of denitrification and anammox.
基金Acknowledgements This research was supported by the National Natural Science Foundation of China (Grant No. 50878180).
文摘Aerobic granules were formed in a conven- tional, continuous flow, completely mixed activated sludge system (CMAS). The reactor was inoculated with seed sludge containing few filaments and fed with synthetic municipal wastewater. The settling time of the sludge and the average dissolved oxygen (DO) of the reactor were 2 h and 4.2 mg. L 1, respectively. The reactor was agitated by a stirrer, with a speed of 250r·min^-1, to ensure good mixing . The granular sludge had good settleability, and the sludge volume index (SVI) was between 50 and 90 mL ·g ^-1. The laser particle analyzer showed the diameter of the granules to be between 0.18 and 1.25 mm. A scanning electron microscope (SEM) investigation revealed the predominance of sphere-like and rod-like bacteria, and only few filaments grew in the granules. The microbial community structure of the granules was also analyzed by polymerase chain reaction-denaturing gradient gel electro- phoresis (PCR-DGGE). Sequencing analysis indicated the dominant species were α, β, and γ-Proteobacteria, Bacteroidetes, and Firmicutes. The data from the study suggested that aerobic granules could form, if provided with sufficient number of filaments and high shear force. It was also observed that a high height-to-diameter ratio of the reactor and short settling time were not essential for the formation of aerobic granular sludge.
基金supported by the National Natural Science Foundation of China (No.NSFC20976069)the Fundamental Research Funds for the Central Universities,China (No.JUSRP111A12)+1 种基金the Higher School Science and Technology Innovation Project of Cultivating the Capital Project,China (No.708048)the Selfdetermined Research Program of Jiangnan University (No.JUSRP11006)
文摘Two identical full-scale biogas-lift reactors treating brewery wastewater were inoculated with different types of sludge to compare their operational conditions, sludge characteristics, and kinetic models at a mesophilic temperature. One reactor (R1) started up with anaerobic granular sludge in 12 weeks and obtained a continuously average organic loading rate (OLR) of 7.4 kg chemical oxygen demand (COD)/(m3.day), COD removal efficiency of 80%, and effluent COD of 450 mg/L. The other reactor (R2) started up with residual activated sludge in 30 weeks and granulation accomplished when the reactor reached an average OLR of 8.3 kg COD/(m^3·day), COD removal efficiency of 90%, and effluent COD of 240 mg/L. Differences in sludge characteristics,biogas compositions, and biogas- lift processes may be accounted for the superior efficiency of the treatment performance of R2 over R1. Grau second-order and modified StoverKincannon models based on influent and effluent concentrations as well as hydraulic retention time were successfully used to develop kinetic parameters of the experimental data with high correlation coefficients (R2 〉 0.95), which further showed that R2 had higher treatment performance than R1. These results demonstrated that residual activated sludge could be used effectively instead of anaerobic granular sludge despite the need for a longer time.
基金This research was supported by the Natural Science Foundation of China(Grant No.50378094).
文摘Shortcut nitrification-denitrification,anaerobic ammonium oxidation(ANAMMOX),and methanogenesis have been successfully coupled in an Expanded Granular Sludge Bed-Biological Aerated Filter(EGSB-BAF)integrated system.As fed different synthetic wastewater with chemical oxygen demand(COD)of 300-1200 mg·L^(-1)and NH_(4)^(+)-N of 30-120 mg·L^(-1)at the outer recycle ratio of 200%,the influence of influent on ANAMMOX in the integrated system was investigated in this paper.The experimental results showed that higher COD concentration caused an increase in denitrification and methanogenesis but a decrease in ANAMMOX;however,when an influent with the low concentration of COD was used,the opposite changes could be observed.Higher influent NH_(4)^(+)-N concentration favored ANAMMOX when the COD concentration of influent was fixed.Therefore,low COD=NH_(4)^(+)-N ratio would decrease competition for nitrite between ANAMMOX and denitrification,which was favorable for reducing the negative effect of organic COD on ANAMMOX.The good performance of the integrated system indicated that the bacterial community of denitrification,ANAMMOX,and methanogenesis could be dynamically maintained in the sludge of EGSB reactor for a certain range of influent.
基金Acknowledgements This work was supported by the National Natural Science Foundation of China (Grant No. 50838005) and National Water Pollution Control and Management Technology Major Projects of China (No. 2009ZX07317-009). The authors are grateful for much assistance with microsensor measurements from Xi'an University Architecture and Technol- ogy.
文摘This paper reports the effects of shear force on anoxic granular sludge in sequencing batch reactors (SBR). The study was carried out in two SBRs (SBR1 and SBR2) in which sodium acetate (200mg COD·L^-1) was used as the sole substrate and sodium nitrate (40 mgNO3-N·L^-1) was employed as the electron acceptor. The preliminary objective of this study was to cultivate anoxic granules in the SBR in order to investigate the effects of shear force on the formation of anoxic granular sludge and to compare the properties of anoxic sludge in the SBR. This study reports new results for the values of average velocity gradient, a measure of the applied shear force, which was varied in the two SBRs (3.79 s^-1 and 9.76 s^-1 for SBR1 and SBR2 respectively). The important findings of this research highlight the dual effects of shear force on anoxic granules. A low shear force can produce large anoxic granules with high activity and poor settling ability, whereas higher shear forces produce smaller granules with better settling ability and lower activity. The results of this study show that the anoxic granulation is closely related to the strength of the shear force. For high shear force, this research demonstrated that: 1) granules with smaller diameters, high density and good settling ability were formed in the reactor, and 2) granular sludge formed faster than it did in the low shear force reactor (41days versus 76 days). Once a steady-state has been achieved, the nitrate and COD removal rates were found to be 98% and 80%, respectively. For low shear force, such as was applied in SBR1, this research demonstrated that: 1) the activity of anoxic granular sludge in low shear force was higher than that in high shear force, 2) higher amount of soluble microbial products (SMPs) were produced, and 3) large pores were observed inside the larger granules,which are beneficial for nitrogen gas diffusion. Electron microscopic examination of the anoxic granules in both reactors showed that the morphology of the granules was ellipsoidal with a clear outline. Coccus and rod-shaped bacteria were wrapped by filamentous bacteria on the surface of granule.