活性污泥体系中聚糖菌的富集与鉴定

活性污泥体系中,聚糖菌(GAOs)在厌氧环境下与聚磷菌(PAOs)形成对底物的竞争关系,对聚糖菌的研究对于优化生物除磷工艺有重要意义。以葡萄糖为惟一碳源,在磷限制条件下,利用特殊运行方式对活性污泥进行驯化培养出了稳定的聚糖菌颗粒污泥,厌氧阶段磷释放量与有机物吸收量浓度(mg/L)比从7.4%下降为0.25%。从培养好的活性污泥反应器中分离获得2株聚糖菌,经菌落PCR和16S rRNA序列分析确定了所得聚糖菌菌株G1和菌株G2分别是枯草芽孢杆菌(Bacillus subtilis)和解鸟氨酸克雷伯氏菌(Klebsiella ornithinolytica)。     

Temperature influence on biological phosphorus removal induced by aerobic/extended-idle regime

Previous researches have demonstrated that biological phosphorus removal (BPR) from wastewater could be driven by the aerobic/extended-idle (A/EI) regime. This study further investigated temperature effects on phosphorus removal performance in six A/EI sequencing batch reactors (SBRs) operated at temperatures ranging from 5 to 30 °C. The results showed that phosphorus removal efficiency increased with temperature increasing from 5 to 20 °C but slightly decreased when temperature continually increased to 30 °C. The highest phosphorus removal rate of 97.1 % was obtained at 20 °C. The biomass cultured at 20 °C contained more polyphosphate accumulating organisms (PAO) and less glycogen accumulating organisms (GAO) than that cultured at any other temperatures investigated. The mechanism studies revealed that temperature affected the transformations of glycogen and polyhydroxyalkanoates, and the activities of exopolyphosphatase and polyphosphate kinase activities. In addition, phosphorus removal performances of the A/EI and traditional anaerobic/oxic (A/O) SBRs were compared at 5 and 20 °C, respectively. The results showed the A/EI regime drove better phosphorus removal than the A/O regime at both 5 and 20 °C, and more PAO and less GAO abundances in the biomass might be the principal reason for the higher BPR in the A/EI SBRs as compared with the A/O SBRs.     

Effect of initial pH control on enhanced biological phosphorus removal from wastewater containing acetic and propionic acids

In the literature most of the studies on the effect of pH on enhanced biological phosphorous removal were conducted with the acetate wastewater, and the pH was controlled during the entire anaerobic and aerobic stages. This paper investigated the influence of anaerobic initial pH control, which will be more practical than the entire process pH control strategy, on enhanced biological phosphorus removal from wastewater containing acetic and propionic acids. Typical pH profile showed that both the initial alkaline and acidic pH tended to neutralize due to the consumption of short-chain fatty acid (SCFA) and intracellular pH regulation by polyphosphate accumulating organisms (PAOs). It was observed that the glycogen degradation and polyhydroxyalkanoates (PHA) accumulation decreased with increasing initial pH, which disagreed with previous reports. In the literature the metabolisms of both glycogen and PHA by PAOs in the acetate wastewater were independent of pH. An anaerobic mechanism model was proposed to explain the intra- and extra-cellular pH buffer nature of PAOs, and to address the reasons for increased polyphosphate degradation and decreased PHA synthesis and glycogen degradation at higher pH. The optimal initial pH for higher soluble ortho-phosphorus (SOP) removal efficiency should be controlled between 6.4 and 7.2. This pH control strategy will be easier to use in practice of wastewater treatment plant.     

基于不同基质的强化生物除磷系统中生化反应机理研究进展

阐述了3种不同基质(乙酸盐、丙酸盐和葡萄糖)在强化生物除磷系统中的生化模型反应机理.重点介绍了聚磷菌(PAOs)、聚糖菌(GAOs)和产乳酸菌在厌氧/好氧条件下对能量及还原力(NADH2)的利用方式;聚-β-羟基链烷酸盐(PHA)的合成方式及存在形式,糖原、乳酸(L型)的代谢途径等.虽然控制基质条件可以实现稳定的强化生物除磷效果,但目前的生化模型并不能完全解释所有的代谢过程,今后要在分离纯种的PAOs及相关生化代谢过程中酶的活动等方面进行深入研究.     

温度对SBR强化生物除磷工艺除磷性能的影响

以厌氧/好氧交替运行的序批式反应器（SBR）为对象,利用荧光原位杂交技术（FISH）,研究了温度（20、25和30℃）对强化生物除磷（EBPR）的影响。结果表明,温度为20℃时,系统的磷去除率高于98%,厌氧释磷速率和好氧吸磷速率分别为55.70 mg P·（g VSS·h）-1和45.16 mg P·（g VSS·h）-1,聚磷菌（PAOs）占总细菌（EUB）的比例达到90%,而聚糖菌（GAO）的比例只有1%;温度升高到25℃后,除磷效果不断降低,释磷速率和吸磷速率逐渐下降,PAOs的比例下降,而聚糖菌（GAOs）的比例不断增加;温度为30℃时,出水水质恶化,磷去除率仅为67%,释磷速率和吸磷速率分别为33.66 mg P·（g VSS·h）-1和17.55 mg P·（g VSS·h）-1,GAOs的比例高达87%,而PAOs的比例仅为5%,在与PAOs的竞争中,GAOs处于优势,导致除磷效果降低。     

温度对生物除磷系统微生物种群关系及动力学的影响

以西安市第三污水处理厂的氧化沟工艺为对象,利用荧光原位杂交技术（FISH）,研究了温度对强化生物除磷系统（EBPR）除磷性能及微生物种群关系的影响。结果表明：当水温低于20 ℃时,厌氧释磷速率和乙酸吸收速率随着温度上升逐渐增加,聚磷菌（PAOs）占总细菌（EUB）的比例也逐渐增加;当水温在20~25 ℃之间,厌氧释磷速率达到最大值,为10.86 mg P·（g VSS·h）-1,乙酸吸收速率随温度升高持续增加,PAOs占EUB的比例也达到最大值,为6.65%;当水温高于25 ℃时,厌氧释磷速率随温度的升高而下降,而乙酸吸收速率则继续增加,聚糖菌（GAOs）的数量已经超过PAOs而成为优势菌,导致处理效果下降。通过对实验数据的统计,得出活性污泥厌氧乙酸吸收速率的温度系数为1.018。     

Functionally relevant microorganisms to enhanced biological phosphorus removal performance at full-scale wastewater treatment plants in the United States

The abundance and relevance ofAccumulibacter phosphatis (presumed to be polyphosphate-accumulating organisms [PAOs]), Competibacter phosphatis (presumed to be glycogen-accumulating organisms [GAOs]), and tetrad-forming organisms (TFOs) to phosphorus removal performance at six full-scale enhanced biological phosphorus removal (EBPR) wastewater treatment plants were investigated. Coexistence of various levels of candidate PAOs and GAOs were found at these facilities. Accumulibacter were found to be 5 to 20% of the total bacterial population, and Competibacter were 0 to 20% of the total bacteria population. The TFO abundance varied from nondetectable to dominant. Anaerobic phosphorus (P) release to acetate uptake ratios (P(rel)/HAc(up)) obtained from bench tests were correlated positively with the abundance ratio of Accumulibacter/(Competibacter +TFOs) and negatively with the abundance of (Competibacter +TFOs) for all plants except one, suggesting the relevance of these candidate organisms to EBPR processes. However, effluent phosphorus concentration, amount of phosphorus removed, and process stability in an EBPR system were not directly related to high PAO abundance or mutually exclusive with a high GAO fraction. The plant that had the lowest average effluent phosphorus and highest stability rating had the lowest P(rel)/HAc(up) and the most TFOs. Evaluation of full-scale EBPR performance data indicated that low effluent phosphorus concentration and high process stability are positively correlated with the influent readily biodegradable chemical oxygen demand-to-phosphorus ratio. A system-level carbon-distribution-based conceptual model is proposed for capturing the dynamic competition between PAOs and GAOs and their effect on an EBPR process, and the results from this study seem to support the model hypothesis.     

The selective role of nitrite in the PAO/GAO competition

Proliferation of Glycogen Accumulating Organisms (GAOs) accounts as one of the major bottlenecks in biological phosphorus removal systems. GAO outcompeting polyphosphate accumulating organisms (PAOs) results in lower P-removal. Thus, finding optimal conditions that favour PAO in front of GAO is a current focus of research. This work shows how nitrite can provide a novel strategy for PAO enrichment. A propionate-fed GAO-enriched biomass (70% Defluviicoccus I, 18% Defluviicoccus II and 10% PAO) was subjected more than 50 d under anaerobic-anoxic conditions with nitrite as electron acceptor. These operational conditions led to a PAO-enriched sludge (85%) where GAO were washed out of the system (<10%), demonstrating the validity of the new approach for PAO enrichment. In addition, the presented suppression of Defluviicocus GAO with nitrite represents an add-on benefit to the nitrite-based systems since the proliferation of non-desirable GAO can be easily ruled out and added to the other benefits (i.e. lower aeration and COD requirements).     

Competition between polyphosphate- and glycogen-accumulating organisms in enhanced-biological-phosphorus-removal systems: effect of temperature and sludge age.

Temperature and sludge age were found to be important factors in determining the outcome of competition between polyphosphate-accumulating organisms (PAOs) and glycogen-accumulating non-polyphosphate organisms (GAOs) and the resultant stability of enhanced-biological-phosphorus removal (EBPR). At 20 degrees C and a 10-day sludge age, PAOs were dominant in an anaerobic/aerobic (A/O) sequencing-batch reactor (SBR), as a result of their higher anaerobic-acetate-uptake rate and aerobic-biomass yield than GAOs. However, at 30 degrees C and a 10-day sludge age, GAOs were able to outcompete PAOs in the A/O SBR because of their higher anaerobic-acetate-uptake rate than PAOs. At 30 degrees C and a 5-day sludge age, GAOs coexisted with PAOs in the A/O SBR, resulting in unstable EBPR performance. At 30 degrees C, reducing the sludge age from 5 to 3 days improved the EBPR efficiency drastically, and the EBPR performance was stable. The maximum specific-anaerobic-acetate-uptake rates of GAO-enriched sludge were affected by temperature with the Arrhenius temperature coefficient theta of 0.042 (degrees C(-1) between 10 and 30 degrees C. The effect of sludge age (5 and 10 days) on the maximum specific-anaerobic-acetate-uptake rates of GAO-enriched activated sludge, however, was not significant. For the PAO-enriched activated sludge, the maximum specific-anaerobic-acetate-uptake rate did not change significantly between 20 and 30 degrees C, but significantly increased from 0.38 to 0.52 mmol-C/ mmol-C/h as the sludge age decreased from 10 to 3 days at 30 degrees C.     

聚糖菌快速富集方法的建立及群落特性分析

为研究聚糖菌(GAOs)模型的群落结构及糖原相关功能基因的表达变化,通过厌氧末排水及磷限制条件的双重策略,建立快速、稳定的聚糖菌模型.应用宏基因组学技术测定了 GAOs体系菌群结构的动态变化,并分析在驯化过程中糖原代谢通路关键功能基因的调节作用.结果表明,经过40d左右的驯化,反应器呈现出典型的GAOs代谢模式,厌氧利...     

不同电子受体对聚磷菌吸磷性能的影响研究

采用人工配水,在厌氧/好氧交替运行的序批式活性污泥反应器(SBR)中,富集了全菌数量80%以上的聚磷菌(Candi-datus Accumulibacter Phosphates)。以此为基础,研究了O2及不同浓度NO3--N、NO2--N对聚磷菌吸磷的影响。结果表明,在一定的条件下,聚磷菌可以NO3--N和NO2--N为电子受体进行缺氧吸磷;NO3--N浓度对聚磷菌的吸磷速率影响很小;聚磷菌可以低质量浓度NO2--N(≤40mg/L)为电子受体,但不能以高质量浓度NO2--N(≥80mg/L)为电子受体,而且高浓度NO2--N对聚磷菌吸磷产生抑制甚至对细菌本身存在毒害;NO2--N为电子受体时,其抑制浓度和污泥本身以及外界条件都存在很大的关系,各个研究结论不尽相同,其影响过程有待进一步的探讨。     

An observation on sludge granulation in an enhanced biological phosphorus removal process

A sequencing batch reactor (SBR) seeded with flocculated sludge and fed with synthetic wastewater was operated for an enhanced biological phosphorus removal (EBPR) process. Eight weeks after reactor startup, sludge granules were observed. The granules had a diameter of 0.5 to 3.0 mm and were brownish in color and spherical or ellipsoidal in shape. No significant change was observed in sludge granule size when operational pH was changed from 7 to 8. The 208-day continuous operation of the SBR showed that sludge granules were stably maintained with a sludge volume index (SVI) between 30 to 55 mL/g while securing a removal efficiency of 83% for carbon and 97% for phosphorus. Fluorescent in situ hybridization (FISH) confirmed the enrichment of polyphosphate accumulating organisms (PAOs) in the SBR. The observations of sludge granulation in this study encourage further studies in the development of granules-based EBPR process.     

低C/N进水城镇污水厂低溶解氧运行效能及微生物变化

低C/N进水污水厂通常需要外加碳源以保障脱氮除磷效能,运行费用高。本研究以摆脱低C/N污水处理对外碳源依赖及降低运营成本为目的,依托山东省某城镇污水厂(A²O工艺)开展基于低DO调控策略的污水厂综合效能研究。结果表明,在长期低DO运行策略下,污水厂逐渐停止外加碳源,在硝化液回流比由370%逐渐降至90%的过程中,TN平均去除率由76.3%提升至82.9%,且除磷效率有所提升。对好氧区氮平衡进行分析发现,在低DO运行条件下,好氧区NH₄⁺-N的转化逐渐以短程硝化和同步反硝化作用为主。微生物宏基因组学结果表明,在低DO条件下,NOB丰度由0.15%下降至0.06%,而具备反硝化功能的PAOs与GAOs的总体丰度由2.06%升高至3.57%。长期低DO、无外加碳源运行条件下,脱氮效能的显著提升主要由于好氧区短程硝化和同步反硝化作用的比例升高所致。低DO对AOB抑制作用小,对NOB抑制作用显著,导致NO₂⁻-N的积累,保障了脱氮效能;PAOs与GAOs利用厌氧期储存的内碳源在好氧区反硝化脱氮,提升了脱氮效能;而内碳源利用率的提升又促进了PAOs的生长,提升了除磷效能。此外,污水厂低DO运行策略下,节省了运营成本,吨水处理费用降低了0.3元。     

基于氧利用速率表征好氧吸磷过程特性

过程监测及调控是废水生物除磷工艺得以长期稳定运行的基础。为了获取氧利用速率（OUR）在生物除磷过程的潜在应用及该过程氧利用特性,开展聚磷菌好氧吸磷过程的呼吸测量实验,分析了OUR曲线变化与磷酸盐浓度变化、胞内聚合物（PHAs）氧化、外源COD存在与否与浓度变化及pH变化之间的响应关系。研究得到如下结果：富集污泥无磷酸盐存在时,体系依然存在明显的氧利用行为,在不吸磷的情况下聚磷菌胞内的PHAs依然会好氧氧化;外源COD的存在将改变PAOs吸磷速率并延长好氧吸磷的结束时间;体系初始pH的不同导致好氧吸磷效率、吸磷过程pH变化趋势及变化程度的不同;不同的初始条件得到不同的OUR。分析证实,利用测量OUR来监测聚磷菌好氧吸磷过程的运行状态是可行的。     

Modeling the aerobic metabolism of polyphosphate-accumulating organisms enriched with propionate as a carbon source.

In enhanced biological phosphorus removal (EBPR) systems, polyphosphate-accumulating organisms (PAOs) are primarily responsible for removing phosphate from wastewater. Propionate is an abundant carbon substrate in many EBPR plants and has been suggested to provide PAOs an advantage over their carbon competitors--the glycogen-accumulating organisms (GAOs). The aerobic metabolism of PAOs enriched with a propionate carbon source is studied in this paper. A metabolic model is proposed and experimentally validated to characterize the aerobic biochemical transformations by PAOs. The model predicts very well the experimental data obtained from the enriched PAO culture through solid-, liquid-, and gas-phase analyses. This model may be combined with previously formulated metabolic models to better describe the biochemical activity of PAOs with acetate and propionate as the primary carbon sources. Furthermore, it can also facilitate the study of the effect of different carbon sources on PAO-GAO competition.     

铁盐化学除磷对活性污泥生物除磷系统的影响

在小型处理系统中生物除磷同步化学除磷简便易行,但加入的除磷剂本身和形成的化学沉淀物的积累可能会对生物系统造成影响。采用序批式生物反应器,对铁盐的2种投加剂量下对活性污泥系统的影响进行了研究。结果表明,向SBR系统中连续投加15 mg·L-1三氯化铁,表观上系统总体的除磷效率较未投加前有一定幅度的提高,活性污泥的沉降性能得到改善,但这削弱了系统的内在生物除磷效力。随着化学除磷的进行,系统污泥胞内PHA含量减少、糖原含量增加;污泥中PAOs相对数量下降而GAOs的相对数量显著增加,优势菌发生演替。结束投加后,污泥的活性可以缓慢恢复。说明该浓度下长时间连续进行同步化学与生物除磷,会对系统造成一定的损害,但这种损害在停止化学除磷后具有一定的可恢复性。而连续投加3 mg·L-1三氯化铁的SBR系统总体除磷效率较未投加前有所提高,且没有对生物除磷系统产生明显抑制作用,能够较好实现化学除磷和生物除磷的协同。     

AOA-SBR系统脱氮除磷性能及PAOs的代谢特性

以SBR反应器为研究对象,采用人工配水,考察在厌氧/好氧/缺氧（AOA）运行方式下系统的脱氮除磷性能,并通过对排水比、好氧停留时间和进水氨氮负荷的调试,优化运行条件。在优化工况下,通过分析典型周期系统中胞内聚合物的形成与转化情况,研究AOA-SBR系统中聚磷菌（PAOs）的代谢特性。结果表明：在AOA-SBR系统中,排水比为50%,好氧停留时间为110 min,进水氨氮负荷为0.20 kg N·（kg MLSS·d）-1是系统的优化工况;厌氧段合成PHA的能量主要来自于聚磷水解,还原力大多来自于糖酵解,只有25% 来自于TCA循环;PHA和糖原是驱动缺氧段反硝化的共同碳源。     

改性生物质炭与聚磷菌联合去除废水中的磷

为了探索改性生物质炭与聚磷菌联合去除废水中磷的效果,将铁改性生物质炭以及聚磷菌投加入序批式废水处理装置中,间隙进水,间隔一定时间测定水中磷浓度随时间变化,并探讨不同水力停留时间、进水COD浓度以及初始磷浓度对联合除磷的影响。研究结果表明,铁改性生物质炭和聚磷菌联用能高效去除废水中磷,在30℃、水力停留时间6 h、COD浓度280 mg·L-1、初始磷浓度5 mg·L-1并且持续曝气条件下,联合除磷效果最优,其磷去除率可达97.94% 。铁改性生物质炭和聚磷菌联合能有效去除废水中磷,为废水除磷提供了新技术。     

单质硫冲击对含硫废水中硫细菌与聚糖菌竞争关系的影响

聚糖菌的富集会与硫循环耦合反硝化除磷系统(denitrifying sulfur conversion-associated enhanced biological phosphorous removal,DS-EBPR)内功能种群微生物——硫细菌发生竞争,从而导致除磷效果波动。因此,首先对母反应器中微生物进行了长期驯化,然后通过向批次实验小反应器中投加单质硫,研究单质硫短期冲击对含硫废水中硫细菌与聚糖菌竞争关系的影响。结果表明,经过长期驯化,在母反应器中发现了硫细菌与聚糖菌共存的现象;而短期冲击实验结果表明,在单个周期反应过程中,虽然单质硫的投加对微生物内源物质(聚羟基脂肪酸酯、糖原)转化量、氮磷去除效果影响不大,但其可以提高硫细菌的活性,增加硫转化量,使得硫细菌在与聚糖菌的竞争中取得优势地位。     

活性污泥系统聚糖菌的富集与菌群多样性分析

在实验室规模续批式反应器(SBR)内,采用人工配水作为系统进水,以乙酸钠为唯一碳源,限制进水磷浓度,调整适宜的运行方式对活性污泥进行驯化,培养富集聚糖菌。90 d的培养过程中,系统的进水COD浓度始终维持在260 mg/L左右,进水PO43--P浓度从20 mg/L逐渐降至2 mg/L,系统除磷能力逐渐丧失。系统内活性污泥的TP/TSS从40.5%降至10.4%,污泥中的糖原/TSS从14.5%增至38.2%,同时COD去除率能保持在90%以上,说明系统中成功富集聚糖菌。采用454高通量测序法分析该系统的活性污泥菌群结构,发现α-proteobacteria,β-proteobacteria,γ-proteobacteria 3类细菌占微生物总量的比例达83.78%。在属的等级上,Tepidicella占活性污泥菌群的比例最高,为20.60%。