亚硝化颗粒污泥中EPS提取方法与组成特性的比较研究

以异养型好氧颗粒污泥(AGS)、普通活性污泥(AS)和厌氧颗粒污泥(An GS)为参照,考察了5种方法对亚硝化颗粒污泥(GNS)中胞外聚合物(EPS)的提取效果,以选出最佳方法,并通过测定蛋白质(PN)/多糖(PS)含量、三维荧光(3D-EEMs)与傅里叶红外(FT-IR)图谱,比较分析了GNS中EPS的组成特性.结果表明,甲醛-Na OH法能在有效保护细胞结构的同时,获得较高的EPS提取效率,而耗时更短的甲醛-热碱法对GNS也同样适用.尽管亚硝化颗粒污泥的EPS总量低于其他污泥样品,但高比例的PS成分有利于自养型微生物的固着生长.从可溶性微生物代谢产物(λEx/λEm=250~310 nm/320~380 nm)和芳香类蛋白质(λEx/λEm=200~250 nm/280~380 nm)所对应的荧光峰强度及红外图谱可知,GNS具有比AS更高的微生物活性和更丰富的结构性蛋白,且EPS组成更为简单.     

Effects of alkalinity on interaction between EPS and hydroxy-aluminum with different speciation in wastewater sludge conditioning with aluminum based inorganic polymer flocculant

Extracellular polymeric substances (EPS) form a stable gel-like structure to combine with water molecules through steric hindrance, making the mechanical dewatering of wastewater sludge considerably difficult. Coagulation/flocculation has been widely applied in improving the sludge dewatering performance, while sludge properties (organic fraction and solution chemistry conditions) are highly changeable and have important effects on sludge flocculation process. In this work, the alkalinity effects on sludge conditioning with hydroxy-aluminum were comprehensively investigated, and the interaction mechanisms between EPS and hydroxy-aluminum with different speciation were unraveled. The results showed that the effectiveness of hydroxy-aluminum conditioning gradually deteriorated with increase in alkalinity. Meanwhile, the polymeric hydroxy-aluminum (Al₁₃) and highly polymerized hydroxy-aluminum (Al₃₀) were hydrolysed and converted into amorphous aluminum hydroxide (Al(OH)₃), which changed the flocculation mechanism from charge neutralization and complexing adsorption to hydrogen bond interaction. Additionally, both Al₁₃ and Al₃₀ showed higher binding capacity for proteins and polysaccharides in EPS than monomeric aluminum and Al(OH)₃. Al₁₃ and Al₃₀ coagulation changed the secondary structure of proteins in EPS, which caused a gelation reaction to increase molecular hydrophobicity of proteins and consequently sludge dewaterability. This study provided a guidance for optimizing the hydroxy-aluminum flocculation conditioning of sludge with high solution alkalinity.     

Evaluation of the secondary structures of protein in the extracellular polymeric substances extracted from activated sludge by different methods

The changes of protein secondary structures in the extracellular polymeric substances(EPS) extracted from activated sludge by four different methods were studied by analyzing the amide I region(1700–1600 cm-1) of the Fourier transform infrared spectra and model protein test. The results showed the molecular weight distribution of organic matter extracted by centrifugation, heating and cation exchange resin(CER) was similar, while the EPS extracted by centrifugation(Control) and CER had similar fluorescent organic matter. The protein secondary structures of extracted EPS by the four methods were different. The similarities of protein secondary structures between the EPS extracted by CER with the Control were the highest among the four extracted EPS. Although the EPS yield extracted by formaldehyde + NaOH method were the highest, its protein secondary structures had the lowest similarity with those extracted by the Control. Additionally, the effects of centrifugation and CER extraction on the secondary structures of bovine serum albumin were also lower than that of other extraction processes. CER enables the second maximum extraction of EPS and maximum retention of the original secondary structure of proteins.     

好氧污泥颗粒化中胞外聚合物（EPS）的动态变化

在SBR中分别运行普通活性污泥和好氧颗粒污泥工艺,考察普通絮体污泥颗粒化过程中EPS的组分变化和分布情况.结果表明,通过减少沉淀时间可以获得质量高的颗粒污泥,污泥系统中的EPS可划分为紧密结合型、松散结合型和溶解性3种;普通污泥期、颗粒污泥初期和颗粒污泥稳定期的EPS含量均以紧密结合型EPS为主,颗粒污泥中总EPS和溶解性EPS含量均高于普通污泥,且颗粒形成初期溶解性EPS增长明显;颗粒污泥中紧密结合型EPS含量相对稳定,松散结合型EPS在不同污泥中含量很低,一个典型反应周期中蛋白质和多糖的变化趋势普遍是先降低后上升,普通污泥和颗粒污泥EPS中蛋白质含量均高于多糖,颗粒形成初期EPS中蛋白质含量有明显上升;普通絮体污泥中EPS和细菌分布均匀,颗粒污泥的表层聚集大量的细菌、内部主要成分是EPS.     

超声波-缺氧/好氧消化过程污泥EPS、酶活性及菌落数的变化研究

为研究超声波促进污泥缺氧/好氧消化的机理,采用最佳超声波参数（超声频率28kHz,超声密度0.1W.mL-1,超声时间15min,超声间隔时间12h）与缺氧/好氧消化衔接进行间歇试验,研究了消化过程中污泥胞外聚合物（EPS）组分（蛋白质、多糖、DNA）和相应溶出物占总量的比例,以及酶活性（蛋白酶,脱氢酶）和菌落数的变化...     

Functions and behaviors of activated sludge extracellular polymeric substances (EPS): a promising environmental interest

Extracellular polymeric substances （EPS） are the predominant constituents of activated sludge and represent up to 80% of the mass of activated sludge. They play a crucial role in the flocculation, settling and dewatering of activated sludge. Furthermore, EPS also show great efficiency in binding heavy metals. So EPS are key factors influencing reduction in sludge volume and mass, as well as activity and utilization of sludge. EPS are of considerable environmental interest and hundreds of articles on EPS have been published abroad, while information on EPS in China is limited. In this paper, results of over 60 publications related to constituents and characteristics of EPS and their influences on flocculation, settling and dewatering of sludge are compiled and analyzed. Metal-binding ability of EPS is also discussed, together with a brief consideration of possible research interests in the future.     

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.     

Effect of Mn2+ on the phosphorus removal and bioflocculation under anoxic condition

Manganese ion (Mn²⁺) generated from metallurgical, steel making and chemical industries enters sewage treatment plants and affects the sludge activity and flocculation. The effect of Mn²⁺ on the removal of chemical oxygen demand (COD) and total phosphorus (TP) and sludge activity were investigated in anoxic zone of an anaerobic/anoxic/oxic (A²O) process. The compositions and structures of extracellular polymeric substances (EPS) were characterized using three-dimensional excitation emission matrix fluorescence spectroscopy (3D-EEM), X-ray photoelectron spectroscopy (XPS) and Fourier transform infrared spectroscopy (FTIR) to reveal the relationship among Mn²⁺, EPS and sludge flocculation.The results showed that low concentration of Mn²⁺ (<5 mg/L) improved removal efficiencies of COD and TP and increased the activity of alkaline phosphatase, acid phosphatase and dehydrogenase. Meanwhile, the addition of Mn²⁺ increased total EPS, sludge contact angle, Zeta potential and sludge particle size, and thus enhanced sludge flocculation. However, high concentration of Mn²⁺ (>10 mg/L) hindered microbial flocculation and reduced removal efficiencies of the pollutants. When Mn²⁺was 5 mg/L, removal efficiencies of COD and TP reached 65% and 90%, respectively. Sludge flocculation was the best and SVI was 70.56 mL/g. The changes of Mn²⁺ concentration caused deviation of groups’ compositions in LB-EPS and TB-EPS, where the main components were always protein (PN) and polysaccharide (PS). The addition of Mn²⁺ resulted in the degradation of humic acids. However, it did not give rise to significant morphology changes of EPS.     

活性污泥胞外聚合物中蛋白质的探针分析方法

应用微相吸附-光谱修正技术(Microsurface adsorption-spectral correction,简称MSASC),利用刚果红(Congo Red,简称CR)探针测定了活性污泥胞外聚合物(Extracellular polymeric substances,简称EPS)中蛋白质(Protein)的含量,与常用的Folin酚法比较,结果一致,蛋白加标回收率为94.5%～107.1%,相对标准偏差(RSD)＜4.5%.对反应机理的分析和产物结合常数的计算结果表明,在pH 2.21时CR-Protein作用符合Langmuir单分子层吸附理论,产物的结合比79∶1,结合常数K=2.17×106,聚集形式为Protein·CR79,摩尔吸收系数ε664r=8.50×105L·mol-1·cm-1.     

Enhanced dewaterability of waste activated sludge by UV assisted ZVI-PDS oxidation

Ultraviolet（UV） assisted zero-valent iron（ZVI）-activated sodium persulfate（PDS） oxidation（UV-ZVI-PDS） was used to treat waste activated sludge（WAS） in this study.The dewaterability performance and mechanism of WAS dewatering were analyzed.The results showed that UV-ZVI-PDS can obtain better sludge dewatering performance in a wide pH range（2.0-8.0）.When the molar ratio of ZVI/PDS was 0.6,UV was 254 nm,PDS dosage was 200 mg/g TS（total solid）,pH was 6.54,reaction time was 20 min,the CST（capillary s...     

Characterization of changes in extracellular polymeric substances and heavy metal speciation of waste activated sludge during typical oxidation solubilization processes

Biopolymer solubilization is considered to be the rate-limiting stage of anaerobic digestion of waste activated sludge(WAS). Oxidation processes have been proven to be effective in disrupting sludge flocs and causing solubilization of the solid biopolymers. In this study,WAS was treated by NaNO2 or H2 O2 oxidation at p H of 2. The changes in extracellular polymeric substances properties and the speciation of heavy metals were investigated. The results revealed that both NaNO2 and H2 O2 treatments were effective in solubilizing organics in WAS, while the conversion of biopolymers in the two treatment processes was different. Free nitrous acid destroyed the gel network structure of EPS, and organic materials were released from the solid phase to the supernatant. Indigenous peroxidase catalyzed H2 O2 to produce hydroxyl radicals which caused significant solubilization of biopolymers, and the protein-like substances were further degraded into micro-molecule polypeptides or amino acids at high dosages of H2 O2. During the oxidation processes, Zn, Cd and Cu, with excellent mobility, tended to migrate to the supernatant, and thus were easy to remove through the liquid–solid separation process. Ni and As showed moderate migration ability, of which the residual fraction tended to transform into reducible and soluble fractions. With poor mobility, Cr and Pb mainly existed in the forms of residual and oxidizable fractions, which were difficult to dissolve and remove from WAS. Both NaNO2 and H2 O2 treatment resulted in the enhancement of sludge solubilization efficiency and heavy metal mobility in WAS, but different heavy metals showed distinct migration and transformation behaviors.     

好氧/厌氧污泥胞外聚合物(EPS)的提取方法研究

采用5种方法对同一污泥在好氧和厌氧条件下的胞外聚合物(EPS)进行了提取试验研究.结果表明,甲醛-NaOH和H2SO4法对EPS的提取产量最高,分别为232·0mg·g~(-1)和159·7mg·g~(-1),且无大量细胞自溶发生,是比较有效的提取方法;与对照方法相比较,阳离子交换树脂(CER)和戊二醛提取法却存在严重缺陷.两种氧环境条件下污泥EPS中蛋白质的含量均最高,占EPS总量的50%~80%;其次为胞外多糖和DNA.污泥在经由好氧环境到厌氧环境的转变过程中,EPS中主要成分蛋白质和DNA含量降低较为明显,其厌氧/好氧状态下的相应含量比值为0·59~0·91.试验中也发现,提取步骤和检测手段对结果的影响最大.     

活性污泥处理偶氮染料废水过程中胞外聚合物特性研究

采用序批式反应器(SBR)考察了偶氮染料浓度对活性污泥的污染物去除性能及胞外聚合物(Extracellular polymeric substances,EPS)的影响.结果表明,COD、染料及营养物的去除率均随着进水染料浓度的增加而下降,但进水染料浓度对EPS的影响却呈现不同的趋势.当进水染料浓度为5 ～40 mg·L-1时,EPS含量随着染料浓度的增加而增加;当进水染料浓度超过40 mg·L-1时,EPS含量却随着染料浓度的增加而减少.染料的加入导致活性污泥EPS中蛋白质的含量增加,且其变化趋势与EPS变化一致;EPS中腐殖酸的浓度低于蛋白质浓度;而EPS中多糖的浓度最低,为8 mg·g-1.三维荧光光谱结果显示,不同染料浓度下EPS荧光吸收峰数量及位置相同,分别为类蛋白峰(λEx/λEm=240 nm/375 ～394 nm)和类富里酸类峰(λEx/λEm =270 nm/410～416 nm),但两个吸收峰的荧光强度不同.     

Insights into sludge granulation during anaerobic treatment of high-strength leachate via a full-scale IC reactor with external circulation system

In this study, a full-scale internal circulation(IC) reactor coupled with an external circulation system was developed to treat high-strength leachate from a municipal solid waste(MSW)incineration plant, in which anaerobic sludge granulation was intensively investigated. Results showed that the IC reactor achieved excellent treatment performance under high organic loading rates(OLR) of 21.06–25.16 kg chemical oxygen demand(COD)/(m3? day). The COD removal efficiency and biogas yield respectively reached 89.4%–93.4% and 0.42–0.50 m3/kg COD.The formation of extracellular polymeric substances(EPS) was closely associated with sludge granulation. Protein was the dominant component in sludge EPS, and its content was remarkably increased from 21.6 to 99.7 mg/g Volatile Suspended Solid(VSS) during the reactor operation. The sludge Zeta potential and hydrophobicity positively correlated with the protein/polysaccharide ratio in EPS, and they were respectively increased from-26.2 m V and 30.35% to-10.6 m V and 78.67%, which was beneficial to microbial aggregation. Three-dimensional fluorescence spectroscopy(3 D-EEM) and Fourier transform infrared spectroscopy(FT-IR)analysis further indicated the importance of protein-like EPS substances in the sludge granulation. Moreover, it was also found that the secondary structures of EPS proteins varied during the reactor operation.     

Occurrence, distribution, and potential influencing factors of sewage sludge components derived from nine full-scale wastewater treatment plants of Beijing, China

Millions of tons of waste activated sludge (WAS) produced from biological wastewater treatment processes cause severe adverse environmental consequences. A better understanding of WAS composition is thus very critical for sustainable sludge management. In this work, the occurrence and distribution of several fundamental sludge constituents were explored in WAS samples from nine full-scale wastewater treatment plants (WWTPs) of Beijing, China. Among all the components investigated, active heterotrophic biomass was dominant in the samples (up to 9478 mg/L), followed by endogenous residues (6736 mg/L), extracellular polymeric substances (2088 mg/L), and intracellular storage products (464 mg/L) among others. Moreover, significant differences (p < 0.05) were observed in composition profiles of sludge samples among the studied WWTPs. To identify the potential parameters affecting the variable fractions of sludge components, wastewater source as well as design and operational parameters of WWTPs were studied using statistical methods. The findings indicated that the component fraction of sewage sludge depends more on wastewater treatment alternatives than on wastewater characteristics among other parameters. A principal component analysis was conducted, which further indicated that there was a greater proportion of residual inert biomass in the sludge produced by the combined system of the conventional anaerobic/anoxic/oxic process and a membrane bioreactor. Additionally, a much longer solids retention time was also found to influence the sludge composition and induce an increase in both endogenous inert residues and extracellular polymeric substances in the sludge.     

Insight into the distribution of metallic elements in membrane bioreactor: Influence of operational temperature and role of extracellular polymeric substances

The distribution of metallic elements in a submerged membrane bioreactor(MBR) was revealed at different temperatures using inductively coupled plasma-optical emission spectrometry(ICP-OES), and the role of extracellular polymeric substances(EPS) was probed by integrating scanning electron microscopy(SEM) with confocal laser scanning microscopy(CLSM) over long-term operation. More metallic elements in the influent were captured by suspended sludge and built up in the fouling layer at lower temperature. The concentration of metallic elements in the effluent was 5.60 mg/L at 10°C operational temperature, far lower than that in the influent(51.35 mg/L). The total contents of metallic elements in suspended sludge and the membrane fouling layer increased to 40.20 and 52.19 mg/g at 10°C compared to 35.14 and 32.45 mg/g at 30°C, and were dominated by the organically bound fraction. The EPS contents in suspended sludge and membrane fouling layer sharply increased to 37.88 and 101.51 mg/g at 10°C, compared to 16.87 and 30.03 mg/g at 30°C. The increase in EPS content at lower temperature was responsible for the deposition of more metallic ions. The strong bridging between EPS and metallic elements at lower temperature enhanced the compactness of the fouling layer and further decreased membrane flux. This was helpful for understanding the mechanism of membrane fouling at different operational temperatures and the role of EPS, and also of significance for the design of cleaning strategies for fouled membranes after long-term operation.     

污泥EPS高值、高效提取与回收技术发展趋势

污泥有机质(特别是胞外聚合物,EPS)高值产品回收将会推动污水/污泥资源化.然而,EPS典型提取方法效率较低、回收产物成分复杂,限制了 EPS目标物质的高效提取、降低了回收物的潜在高值应用.为此,探究高值、高效EPS提取/回收技术极为重要.这就需要改变目前典型的"笼统"EPS提取方法,将"胞外多糖"和"胞外蛋白"这样的...     

Effect of proteins, polysaccharides, and particle sizes on sludge dewaterability

Four batch experiments of hydrolysis and acidification were carried out to investigate the distributions of proteins (PN) and polysaccharides (PS) in the sludge, the PN/PS ratio, the particle sizes, and their relationship with sludge dewaterability (as determined by capillary suction time, CST). The sludge flocs were stratified through centrifugation- and ultrasound-based method into four fractions: (1) slime, (2) loosely bound extracellular polymeric substances (LB-EPS), (3) tightly bound EPS (TB-EPS), and (4) pellet. The results showed that PN was mainly partitioned in the pellet (80.7%) and TB-EPS (9.6%) fractions, while PS distributed evenly in the four fractions. During hydrolysis and acidification, PN was transferred from the pellet and TB-EPS fractions to the slime fraction, but PS had no significant transfer trends. The mean particle sizes of the sludge flocs decreased with hydrolysis and acidification. The pH had a more significant influence on the dewaterability of sludge flocs than temperature. Sludge dewaterability during hydrolysis and acidification processes greatly deteriorated from 9.7 s at raw sludge to 340–450 s under alkaline conditions. However, it was just slightly increased under acidic conditions. Further investigation suggested that CST was a ected by soluble PN, soluble PN/PS, and particle sizes of sludge flocs, but was a ected slightly by total PN, PS, or PN/PS in the whole sludge flocs and other fractions (except slime).     

Effect of sludge retention time on sludge characteristics and membrane fouling of membrane bioreactor

Three identical membrane bioreactors (MBRs) were operated over 2 years at different sludge retention time (SRT) of 10 d, 40 d and no sludge withdrawal (NS), to elucidate and quantify the effect of SRT on the sludge characteristics and membrane fouling. The hydraulic retention times of these MBRs were controlled at 12 h. With increasing SRT, the sludge concentrations in the MBRs increased, whereas the ratio of volatile suspended solid to the total solid decreased, and the size of sludge granule diminished in the meantime. A higher sludge concentration at long SRT could maintain a better organic removal efficiency, and a longer SRT was propitious to the growth of nitrifiers. The performance of these MBRs for the removal of COD and NH₄⁺-N did not change much with different SRTs. However, the bioactivity decreased as SRT increase. The measurement of specific oxygen uptake rates (SOUR) and fluorescence in situ hybridization (FISH) with rRNA-targeted oligonucleotide probes testified that SOUR and the proportion of the bacteria-specific probe EUB338 in all DAPI-stainable bacteria decreased with increasing SRT. The concentrations of total organic carbon, protein, polysaccharides and soluble extracellular polymeric substance (EPS) in the mixed liquor supernatant also decreased with increasing SRT. The membrane fouling rate was higher at shorter SRT, and the highest fouling rate appeared at a SRT of 10 d. Both the sludge cake layer and gel layer had contribution to the fouling resistance, but the relative contribution of the gel layer decreased as SRT increase.     

三维荧光光谱法研究四环素与活性污泥EPS的相互作用

用三维荧光光谱仪研究了活性污泥EPS(Extracellular polymeric substances)和四环素的相互作用。活性污泥EPS中发现2种类蛋白荧光峰A(Ex/Em=225/340)和B(Ex/Em=280/338),其荧光强度随着盐酸四环素的加入而显著降低,峰A和盐酸四环素结合生成了不发荧光的物质,而峰B和盐酸四环素结合生成的物质在Ex/Em=295/416 nm处发荧光。峰A和峰B与盐酸四环素的相互作用适合用修正后的Stern-Volmer和Hill模型拟合,盐酸四环素和峰A的结合点位数、结合常数分别为1.33、7.02,大于和峰B的结合点位数和结合常数,说明盐酸四环素和荧光峰A的结合比较稳定。猝灭速率常数kq远远大于最大扩散碰撞猝灭速率常数(2.0×1010L(/mol.s)),表明峰A和峰B和盐酸四环素相互作用的荧光猝灭主要是静态猝灭过程。因此土壤和水环境中大量存在EPS会影响盐酸四环素在环境中的迁移转化。