Performance of Anammox granular sludge bed reactor started up with nitrifying granular sludge

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.

Production and application of anaerobic granular sludge produced by landfill

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.

Comparison of membrane fouling during short-term filtration of aerobic granular sludge and activated sludge

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.

Study of 4-t-octylphenol degradation and microbial community in granular sludge

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.

Community analysis of ammonia and nitrite oxidizers in start-up of aerobic granular sludge reactor

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.

Re-activation characteristics of preserved aerobic granular sludge

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.

Eubacteria and Archaea community of simultaneous methanogenesis and denitrification granular sludge

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%.

Influence of Chemical Oxygen Demand Concentrations on Anaerobic Ammonium Oxidation by Granular Sludge From EGSB Reactor

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.

Stability of expanded granular sludge bed process for terylene artificial silk printing and dyeing wastewater treatment

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.

Molecular characterization of bacterial community in aerobic granular sludge stressed by pentachlorophenol

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...

Niche Differentiation of Phenol-Degrading Microorganisms in UASB Granular Sludge as Revealed by Fluorescence in situ Hybridization

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.

Experimental Study on Granular Sludge Cultivation Based on IC Reactor

[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.

Screening and identification of functional bacterial attachment genes in aerobic granular sludge

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.

Effect of seed sludge on characteristics and microbial community of aerobic granular sludge

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.

Biodegradation of aniline by Candida tropicalis AN1 isolated from aerobic granular sludge

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.

Bamboo charcoal addition enhanced the nitrogen removal of anammox granular sludge with COD:Performance,physicochemical characteristics and microbial community

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.

Identical full-scale biogas-lift reactors(BLRs) with anaerobic granular sludge and residual activated sludge for brewery wastewater treatment and kinetic modeling

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.

Influence of influent on anaerobic ammonium oxidation in an Expanded Granular Sludge Bed-Biological Aerated Filter integrated system

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.

Performance and microbial diversity of an expanded granular sludge bed reactor for high sulfate and nitrate waste brine treatment

The disposal of waste brines has become a major challenge that hinders the wide application of ion- exchange resins in the water industry in recent decades. In this study, high sulfate removal efficiency （80%-90%） was achieved at the influent sulfate concentration of 3600 mg/L and 3% NaC1 after 145 days in an expanded granular sludge bed （EGSB） reactor. Furthermore, the feasibility of treating synthetic waste brine containing high levels of sulfate and nitrate was investigated in a single EGSB reactor during an operation period of 261 days. The highest nitrate and sulfate loading rate reached 6.38 and 5.78 kg/（m3-day） at SO42--S/NO3-N mass ratio of 4/3, and the corresponding removal efficiency was 99.97% and 82.26% at 3% NaC1, respectively. Meanwhile, 454-pyrosequencing technology was used to analyze the bacterial diversity of the sludge on the 240th day for stable operation of phase X. Results showed that a total of 9194 sequences were obtained, which could be affiliated to 14 phyla, including Proteobacteria, Firmicutes, Chlorobi, Bacteroidetes, Synergistetes and so on. Proteobacteria （77.66%） was the dominant microbial population, followed by Firmicutes （12.23%） and Chlorobi （2.71%）.

Formation process and properties of hydrogen-producing granular sludge in UASB reactor

The granular sludge of microbial fermentation systems includes various biomass-degrading enzymes and different microflora,which have significant impacts on the degradation of biomass and the stability of the system.An up-flow anaerobic sludge blanket(UASB)reactor was used to grow hydrogen-producing granular sludge.The results showed that the formation of the granular sludge underwent four stages,i.e.,flocculation of the sludge,formation of the flocculent sludge,swelling of the flocculent sludge,and formation of the granular sludge.The formed granular sludge mostly had regular spherical and ellipsoidal shapes with a fractal dimension of 2.08±0.4;the settling velocities were 0.84 cm/s to 1.96 cm/s in water,the porosity was 0.67-0.95.The shear sensitivity(Kss)of the granular sludge was 0.1152.The granular sludge had a culture cycle of approximately 70 d and a hydrogen yield of 1.09 mol H2/mol glucose.