Excitation-emission matrix (EEM) fluorescence spectroscopy for characterization of organic matter in membrane bioreactors: Principles,methods and applications

• Principles and methods for fluorescence EEM are systematically outlined. • Fluorophore peak/region/component and energy information can be extracted from EEM. • EEM can fingerprint the physical/chemical/biological properties of DOM in MBRs. • EEM is useful for tracking pollutant transformation and membrane retention/fouling. • Improvements are still needed to overcome limitations for further studies. The membrane bioreactor (MBR) technology is a rising star for wastewater treatment. The pollutant elimination and membrane fouling performances of MBRs are essentially related to the dissolved organic matter (DOM) in the system. Three-dimensional excitation-emission matrix (3D-EEM) fluorescence spectroscopy, a powerful tool for the rapid and sensitive characterization of DOM, has been extensively applied in MBR studies; however, only a limited portion of the EEM fingerprinting information was utilized. This paper revisits the principles and methods of fluorescence EEM, and reviews the recent progress in applying EEM to characterize DOM in MBR studies. We systematically introduced the information extracted from EEM by considering the fluorescence peak location/intensity, wavelength regional distribution, and spectral deconvolution (giving fluorescent component loadings/scores), and discussed how to use the information to interpret the chemical compositions, physiochemical properties, biological activities, membrane retention/fouling behaviors, and migration/transformation fates of DOM in MBR systems. In addition to conventional EEM indicators, novel fluorescent parameters are summarized for potential use, including quantum yield, Stokes shift, excited energy state, and fluorescence lifetime. The current limitations of EEM-based DOM characterization are also discussed, with possible measures proposed to improve applications in MBR monitoring.     

Characterization of the dissolved organic matter in sewage effluent of sequence batch reactor: the impact of carbon source

Dissolved organic matter (DOM) transformation in sequence batch reactor (SBR) fed with carbon sources of different biodegradability was investigated. During the biologic degradation process, the low molecular weight (MW) fraction (< 1 kDa) gradually decreased, while the refractory compounds with higher aromaticity were aggregated. Size exclusion chromatography (SEC) and fluorescence of excitation emission matrices (EEM) demonstrated that more biopolymers (polysaccharides or proteins) and humic-like substances were presented in the extracellular polymeric substance (EPS) extracted from the SBR fed with sodium acetate or glucose, while the EPS from SBR fed with slowly biodegradable dissolved organic carbon (DOC) substratestarch had relatively less biopolymers. Comparing the EfOM in sewage effluent of three SBRs, the effluent from SBR fed with starch is more aromatic. Organic carbon with MW>1 kDa as well as the hydrophobic fraction in DOM gradually increased with the carbon sources changing from sodium acetate to glucose and starch. The DOC fractionation and the EEM all demonstrated that EfOM from the effluent of the SBR fed with starch contained more fulvic acid-like substances comparing with the SBR fed with sodium acetate and glucose.     

Comparative effects of different maize straw returning modes on soil humus composition and humic acid structural characteristics in Northeast China

In order to evaluate the effects of different maize straw returning modes on humus composition and humic acid (HA) structural, a field experiment was designed to have five treatments, i.e. Treatment CK (no straw returned), Treatment EIS (straw incorporated evenly into the soil using the crashing-ridging technique), Treatment SP (straw plowed into the soil), Treatment SM (straw returned as mulch) and Treatment SG (straw returned as pellets). Our results showed that Treatment EIS significantly increased the content of soil organic carbon (SOC), soil humus substances (HEC), soil humic acid carbon (HAC) and fulvic acid carbon (FAC) by 27.8%, 47.6%, 63.3% and 33.8%, respectively, as compared with CK. Among all the straw returning treatments, Treatment EIS was the most significant in effect of increasing the content of HEC, HAC and FAC. Moreover, Treatment EIS altered the composition of humus more significantly than all the other treatments, by increasing the proportion of alkyl C and the ratio of aliphatic C/aromatic C. These results suggest that straw returning accelerated the accumulation of soil organic C and various components of humus, significantly improved the structure of soil HA, with the practice of EIS in particular.     

小球藻来源溶解有机质的光化学降解特性

利用三维荧光光谱-平行因子分析(EEM-PARAFAC)手段结合吸收光谱分析,研究了小球藻指数期和稳定期培养液中溶解有机物(DOM)在秋季天然太阳辐射作用下的光降解动力学特征.结果表明,小球藻生长过程中除能产生短波激发类腐殖质组分C1(其荧光峰的激发/发射波长位置为240,335 nm /406 nm)及类蛋白质组分C3(225,275 nm/334 nm)外,还会形成长波激发类腐殖质组分C2(260,395 nm /502 nm),表明C2组分并非仅有传统认为的陆源属性,同时也具有自生源属性.稳定期培养液的吸收光谱在250~300 nm范围内出现的肩峰,可用于指示水环境中现场自生源的贡献.不同生长时期培养液中DOM的吸收系数和荧光组分的降解动力学都符合一级反应方程.稳定期类腐殖质荧光组分(C1和C2)的光降解程度略高于指数期,但指数期类色氨酸组分(C3)的光降解程度略高于稳定期.经太阳辐照6 d后,小球藻培养液的吸收系数a₃₅₀及各荧光组分的平均损失率分别达到83.0%、84.0%、64.8%和80.0%,对应的半衰期只有1.6~5.0 d,揭示出藻类自生来源的DOM具有很强的光化学降解活性.     

A/O-MBR处理餐饮废水过程中DOM特性解析

借助凝胶过滤色谱(GFC)分子量测定技术和三维荧光(EEM)光谱技术,对厌氧微网与好氧平板膜-生物反应器(A/O-MBR)联合处理餐饮废水过程中各阶段的溶解性有机物(DOM)及污泥胞外聚合物(EPS)的性质进行研究。分子量研究表明:好氧段污泥滤液的GFC谱图中在10 min以前有大分子物质析出,分析是微生物代谢产物;且随着处理过程的进行水样DOM中的分子量分布范围不断变宽。EEM光谱研究表明:进水、好氧段出水、好氧段污泥滤液的DOM中的2种主要荧光物质为高激发波长类色氨酸和可见区类富里酸,而厌氧段出水中仅有高激发波长类色氨酸,且峰强较强。同时,厌氧段出水中在形成可见区类富里酸的区域内有成峰趋势,分析是由较强的类色氨酸峰的掩蔽作用造成的。好氧段污泥EPS中存在3个明显的荧光峰,指示了色氨酸和腐殖酸的存在。     

Multivariate curve resolution of multidimensional excitation-emission quenching matrices of a Laurentian soil fulvic acid

Fluorescence excitation–emission matrices (EEM) of aqueous solutions of Laurentian soil fulvic acid (LFA) at three concentrations (50, 75 and 100 mg/l) were obtained at two pH values (pH = 4.0 and 6.0) and as function of the Cu(II) ion concentration. The presence of Cu(II) ion provokes quenching of the intrinsic LFA fluorescence due to complex formation. Multivariate curve resolution (MCR-ALS) was used to successfully decompose single EEM into excitation and emission spectra for the detected components. Moreover, multidimensional (up to six dimensions) data matrices were generated by adding EEM collected as function of the LFA and Cu(II) concentrations and pH. MCR-ALS was able to resolve the excitation and emission spectra from these multidimensional data matrices given further information about the spectral variation profiles induced by the experimental factors. Conditional stability constants (log K_LFACu) were calculated from the quenching profiles observed as function of the Cu(II) concentration, as well as, their trends as function of pH and LFA concentration were obtained – average (and standard deviation) of log K_LFACu = 4.6 ± 0.2. This EEM/MCR-ALS methodology constitutes a new tool for the study of natural organic matter under varying experimental conditions that characterize natural environmental systems.     

Degradation of dissolved organic matter in effluent of municipal wastewater plant by a combined tidal and subsurface flow constructed wetland

Dissolved organic matter (DOM) is an important constituent of wastewater treatment plant (WWTP) effluent. A novel combined tidal and subsurface flow constructed wetland (TF-SSF-CW) of 90 L was constructed for a ten-month trial of advanced treatment of the WWTP effluent. Excitation emission matrix (EEM) fluorescence spectroscopy, parallel factor (PARAFAC) analysis and a two end-member mixing model were employed to characterize the composition and removal process of the effluent DOM (EfOM) from the WWTP. The results showed that the TF-SSF-CW performed an efficient EfOM removal with dissolved organic carbon (DOC) removal rate of 88% and dissolved organic nitrogen (DON) removal rate of 91%. Further analysis demonstrated that the EfOM consisted mainly of two protein moieties and two humic-like groups; protein moieties (76%) constituted the main content of EfOM in raw water and humic-like groups (57%) became the dominating contributor after treatment. The EfOM from the WWTP was mainly of aquatic bacterial origin and evolved to a higher proportion of terrigenous origin with higher humification in the TF-SSF-CW effluent. A common controlling treatment-related factor for determining the concentrations of the same kind of substances (protein groups or humic-like groups) was revealed to exist, and the ratio of removal rates between the same substances in treatment was calculated. Our study demonstrates that the TF-SSF-CW can be a novel and effective treatment method for the EfOM from WWTPs, and is helpful for understanding of the character of EfOM in wetland treatment.     

巢湖二氧化碳排放特征及其潜在影响因素

为探讨浅水湖泊CO2排放的时空格局及与CDOM来源组成潜在关联机制,于1月(枯水期)、4月(平水期)、7月(丰水期)不同水文情景下富营养浅水巢湖进行野外观测,采用扩散系数—顶空瓶法观测表层水体CO2浓度(cCO2)和通量(FCO2),并探讨CO2排放的潜在驱动因素,尤其是对CDOM来源组成的响应机制.结果表明,巢湖全湖...     

岩溶区水库冬季溶解有机质组成特征及来源:以桂林五里峡水库为例

近年来,三维荧光技术被广泛应用于研究天然水体中溶解有机质(DOM)的物理化学特性.为了解岩溶区水库水体溶解有机质的特征,利用三维荧光光谱并结合平行因子分析法研究桂林五里峡水库冬季水体中DOM分布特征及其来源,探讨其水化学特征以及初步估算DOM荧光组分的相对比例.结果表明,五里峡水库水体中的Ca~(2+)、HCO_3~-为主要阳阴离子,水体水化学主要受碳酸盐风化作用影响,DOC质量浓度占TOC质量浓度的91%;五里峡水库水体DOM在冬季主要由类富里酸、类腐殖酸和类色氨酸组成;荧光指数、腐殖化指标、自生源指标和新鲜度指数显示五里峡水库水体DOM在冬季整体呈现弱腐殖质化特征,自生源特征明显,微型光合生物对水体中DOM的形成起到至关重要的作用,说明水生微型生物的光合作用在岩溶碳循环中扮演着重要的角色.     

重要饮用水源地天目湖水库有色可溶性有机物来源与组成特征

天目湖作为重要集中式饮用水源地,水体水质变化会影响其生态系统服务功能.有色可溶性有机物（CDOM）是溶解性有机物的重要组成部分,其来源与组成直接影响水处理工艺与出水品质,因而研究天目湖CDOM来源及时空分异规律对其水质供应安全及生态系统功能维护有着重要的作用.基于2017年逐月野外采样数据,运用平行因子分析法（PARAFAC）等对CDOM光谱数据进行分析,揭示了天目湖CDOM来源和组成的空间及季节变化特征.PARAFAC结果表明,天目湖CDOM库中微生物作用类腐殖酸组分C1相对丰度最高（44.2%±9.8%）,其次为类色氨酸组分C2（29.2%±4.3%）和类酪氨酸组分C3（17.2%±13.1%）,陆源类腐殖酸组分C4相对丰度最低（9.4%±2.4%）.时空分布特征及主成分分析结果表明天目湖河口区CDOM丰度a（254）、C1和C2组分显著高于下游湖区,而光谱斜率S_275-295则显著低于下游湖区（t-test,P<0.05）,意味着入湖河流输入造成天目湖河口区CDOM腐殖质化程度及相对分子量的升高.夏、秋季节的a（254）、C1、C2和C4组分荧光强度显著高于冬、春季节（t-test,P<0.05）.结果表明不同季节对CDOM组成的影响不仅要考虑降雨量和径流输入的差异,还应综合考虑水体温度、热分层和浮游植物生物量以及光和微生物降解对CDOM的矿化作用.