Comparing results of cultured and uncultured biological methods used in biological phosphorus removal

Increasing attention has been paid to phosphate-accumulating organisms(PAOs)for their important role in biological phosphorus removal.In this study,microbial communities of PAOs cultivated under different carbon sources(sewage,glucose,and sodium acetate) were investigated and compared through culture-dependent and culture-independent methods,respectively.The results obtained using denaturing gradient gel electrophoresis(DGGE)of polymerase chain reaction-amplified 16S rDNA fragments revealed that the diversity of bacteria in a sewage-fed reactor(1#)was much higher than in a glucose-fed one(2#)and a sodium acetate-fed one(3#);there were common PAOs in three reactors fed by different carbon sources.Five strains were separated from three systems by using a phosphate- rich medium;they were from common bacteria isolated and three isolates could not be found in DGGE profile at all.Two isolates had good phosphorus removal ability.When the microbial diversity was studied,the molecular biological method was better than the culture-dependent one.When phosphorus removal characteristics were investigated,culture-dependent approach was more effective. Thus a combination of two methods is necessary to have a comprehensive view of PAOs.     

Role of extracellular exopolymers on biological phosphorus removal

Three sequencing batch reactors supplied with different carbon sources were investigated. The system supplied with glucose gained the best enhanced biological phosphorus removal although all of the three reactors were seeded from the same sludge. With the measurement of poly-β-hydroxyalkanoate （PHA） concentration, phosphorus content in sludge and extracellular exopolymers （EPS） with scanning electron microscopy （SEM） combined with energy dispersive spectrometry （EDS）, it was found that the biosorption effect of EPS played an important role in phosphorus removal and that the amount of PHA at the end of anaerobic phase was not the only key factor to determine the following phosphorus removal efficiency.     

溶解氧对好氧/延长闲置SBR除磷性能的影响

以合成废水为研究对象,乙酸钠为外加碳源,考察不同溶解氧(DO)浓度下好氧/延长闲置(O/EI)序批式反应器的除磷效果,并通过分析典型周期内磷元素及微生物体内各储能物质的变化,探究DO浓度对O/EI工艺除磷性能的影响机制.结果表明,低DO浓度(1 mg·L-1)条件下,O/EI系统具有良好的除磷效果,除磷率高达96%,单位污泥除磷量为5.02 mg·g-1;而当DO浓度较高(4 mg·L-1)时,反应器内磷的去除率降至50%,单位污泥除磷量仅2.81 mg·g-1.研究表明,在DO浓度为1 mg·L-1时,微生物能合成较多聚羟基脂肪酸酯(PHAs),糖原的合成及利用较少,系统好氧吸磷量远高于其他反应器,并在闲置期释放出更多聚磷酸盐.可见,DO可通过影响微生物体内PHAs和糖原的合成及转化,闲置期释磷,好氧前期释磷及好氧吸磷,进而影响系统的除磷性能.     

Effects of idle time on biological phosphorus removal by sequencing batch reactors

Three identical sequencing batch reactors （SBRs） were operated to investigate the effects of various idle times on the biological phosphorus （P） removal. The idle times were set to 3 hr （R1）, 10 hr （R2） and 17 hr （R3）. The results showed that the idle time of a SBR had potential impact on biological phosphorus removal, especially when the influent phosphorus concentration increased. The phosphorus removal efficiencies of the R2 and R3 systems declined dramatically compared with the stable R1 system, and the Prelease and P-uptake rates of the R3 system in particular decreased dramatically. The PCR-DGGE analysis showed that uncultured Pseudomonas sp. （GQ183242.1） and β-Proteobacteria （AY823971） were the dominant phosphorus removal bacteria for the R1 and R2 systems, while uncultured γ-Proteobacteria were the dominant phosphorus removal bacteria for the R3 system. Glycogen-accumulating organisms （GAOs）, such as uncultured Sphingomonas sp. （AM889077）, were found in the R2 and R3 systems. Overall, the R1 system was the most stable and exhibited the best phosphorus removal efficiency. It was found that although the idle time can be prolonged to allow the formation of intracellular polymers when the phosphorus concentration of the influent is low, systems with a long idle time can become unstable when the influent phosphorus concentration is increased.     

不同碳源驯化除磷污泥的除磷效果及菌群结构分析

采用SBR反应器,分别以乙酸钠、甘油、丙酸钠为单一碳源,在严格厌氧/缺氧条件下驯化培养反硝化除磷污泥,考察不同碳源对反硝化除磷效果的影响,并采用高通量测序技术研究了不同碳源除磷污泥的菌群结构,分析其中除磷菌所占比例. 结果表明,以乙酸钠、甘油、丙酸钠为碳源的各系统出水中,ρ(TP)平均值分别为0.79、0.98、0.29 mg/L,TP去除率分别为82.5%、79.2%、93.4%. 取自污水厂的种泥微生物多样性最高;其次为以甘油驯化的污泥和以乙酸钠与丙酸钠培养的污泥,二者表现出相似的多样性与菌群结构. 各反应器中的污泥在“纲”与“目”分类级别上分别均以β-Proteobacteria与Rhodocyclales占主导. 稳定期乙酸钠、甘油、丙酸钠为碳源的系统中的除磷菌所占比例分别为9.5%、8.0%、41.5%,以丙酸钠为碳源的系统中除磷菌所占比例最高. 对于厌氧/缺氧系统,与以乙酸钠、甘油相比,丙酸钠为碳源时系统的除磷效果更好,并且有利于除磷菌的富集.      

丙酸的加入对厌氧-低氧同时生物除磷脱氮系统的影响

2个实验室规模的序批式反应器（SBRs）在厌氧-低氧（0.15～0.45 mg·L^-1）条件下运行,以比较丙酸的加入对同时生物除磷脱氮系统的影响.结果表明,无论是丙酸与乙酸的混合酸（碳摩尔比为1.5/1）作为碳源（SBR1）,还是乙酸作为单独碳源（SBR2）,系统都发生同步硝化反硝化和磷的去除（SNDPR）,并且氨氮被全部氧化,系统中没有亚硝酸盐的大量累积.与SBR2相比,SBR1中厌氧阶段磷释放量少,聚羟基戊酸（PHV）合成量高,好氧末磷剩余量少,硝态氮累积少,因此SBR1中总氮和总磷的去除率（分别为68%和95%）比SBR2（分别为51%和92%）高,加入丙酸有助于SNDPR系统保持较好的除磷、脱氮效果.     

亚硝酸盐不同生成方式对短程硝化反硝化除磷颗粒系统的影响

为了探究亚硝酸盐生成方式对短程硝化反硝化除磷颗粒系统的影响,采用2组同规格SBR反应器分别在连续和间歇曝气方式下使亚硝酸盐连续生成和间歇生成,考察其运行过程中脱氮除磷效果、污泥物理特性和微生物群落结构.结果表明,亚硝酸盐间歇生成后随即消耗,具有更好和更稳定的脱氮除磷性能,特别在TN去除上,第72 d后TN平均去除率为92.07%.碳源利用效率（以P/COD计）集中在0.21~0.22mg·mg^-1,碳源利用充分,进一步促进反硝化除磷.颗粒粒径分布集中,大小均匀,具有规则的形状和清晰的边界.微生物群落分析表明,亚硝酸盐间歇生成的系统微生物群落丰富度和多样性更高,同时富集了更多DPAOs菌属（Dechloromonas和Pseudomonas）,与Nitrosomonas共同作用使短程硝化与反硝化除磷达到动态平衡,实现系统稳定运行.     

游离亚硝酸对好氧/延长闲置序批式反应器除磷性能的影响

以合成废水为研究对象,在好氧/延长闲置序批式反应器中研究不同浓度游离亚硝酸对系统除磷性能的影响.通过分析闲置期聚磷水解及微生物体内各储能物质的变化及不同反应器的除磷效果,探究其可能存在的影响机理.结果表明,游离亚硝酸浓度为5.13×10-5mg·L-1时对反应器除磷效果及沉降能力抑制不明显,而当游离亚硝酸浓度达到5.13×10-4mg·L-1时,磷的平均去除率降至40.5%,反应器中出现严重的污泥膨胀,污泥沉降指数值最高达310 mL·g-1.典型周期分析表明,游离亚硝酸可影响聚磷菌对碳源的吸收和胞内聚羟基脂肪酸酯的合成,进而影响好氧吸磷及闲置期聚磷水解.恢复试验表明,当游离亚硝酸浓度高于2.57×10-4mg·L-1时,撤销影响后系统的除磷能力和沉降性能虽有明显回升,但无法恢复至初始水平.     

好氧段碳源浓度对同步去除和富集磷酸盐生物膜的影响

利用聚磷菌以循环交替O/A模式运行,对生活污水处理厂的主流工艺中实现磷酸盐的同步去除和富集,探究了好氧段碳源浓度对聚磷生物膜去除和富集磷酸盐性能以及生物膜中微生物种群结构的影响.结果表明,好氧COD质量浓度从200 mg·L~(-1)降低到0 mg·L~(-1),吸磷速率提升1. 29倍,出水磷质量浓度稳定在0. 5 mg·L~(-1)以下;释磷速率提升3. 56倍,富集液磷酸盐质量浓度从27. 125 mg·L~(-1)升高到55. 91 mg·L~(-1).微生物群落变化中,鉴定为聚磷菌的变形菌门(Proteobacteria)的含量增加约2倍,红环菌科(Rhodocyclaceae)和厌氧绳菌科(Anaerolineaceae)的富集效果分别提高了2. 28和5倍.降低好氧段碳源浓度,有利于聚磷菌的筛选和富集,强化了好氧段磷酸盐的去除以及厌氧段磷酸盐的释放,获得了更高的磷酸盐富集液,并且为以资源回收为目的的未来城市污水处理厂提供降低好氧段碳源需求的理论基础.     

溶解氧对活性污泥胞内贮存物和除磷性能的影响

分别以乙酸钠和葡萄糖为碳源,通过间歇实验,考察不同溶解氧(DO)浓度对厌氧-缺氧-好氧序批式反应器中活性污泥胞内贮存物聚羟基烷酸酯(PHA)、糖原和系统除磷性能的影响。实验表明:DO对以乙酸钠为碳源培养的胞内贮存物PHA和糖原影响较大,并影响污泥的除磷性能;对以葡萄糖为碳源培养的胞内物质影响较小,可能是因为细胞内存在其他类型的胞内贮存物。     

不同碳源对EBPR启动期聚磷菌的影响研究

以实验室序批式反应器(SBR)为强化生物除磷工艺(EBPR)载体,接种具有初步除磷功能的污泥后,以乙酸∶丙酸=1∶1(按各自折算的COD计)为混合碳源(以下简称混酸),厌氧初始pH 7.6±0.1,富集聚磷菌(PAO)。启动30 d后,EBPR反应器中为PAO和聚糖菌(GAO)的混合菌属,此时从反应器中取泥样进行批式试验,分别考察乙酸、丙酸及混酸对聚磷菌的富集和厌氧释磷的影响。结果表明:在EBPR启动期内,乙酸作为单一碳源时释磷量最大,但混酸碳源释磷效率最高,最有利于PAO富集;丙酸作为单一碳源时降解率最大而释磷量最小,不适合EBPR启动期的PAO富集。     

胞外聚合物磷酸盐形态对生物除磷过程的影响研究

以不同DO条件下污泥龄(SRT)分别为10 d和30 d的两组实验室A/O-SBR反应器活性污泥为研究对象,探讨了胞外聚合物(EPS)磷酸盐形态对生物除磷过程的影响.结果表明,污泥絮体中的磷主要分布于EPS中,PO3-4-P和聚磷酸盐(Poly-P,包括低分子量聚磷酸盐LMW PolyP和高分子量聚磷酸盐HMW Poly-P)是EPS磷的主要形态;EPS对生物除磷的影响明显大于细菌细胞,EPS磷的厌氧降低量和好氧升高量为胞内磷变化量的2.8~6.4倍.EPS中的LMW Poly-P和HMW Poly-P含量均表现厌氧降低和好氧升高的变化规律;对于相同SRT的污泥,中DO(2.5~3.5 mg·L-1)条件较低DO(0.7~1.0 mg·L-1)条件下EPS的LMW Poly-P和HMW Poly-P有更大的厌氧降低量和好氧升高量,对应着更明显的生物除磷过程,说明EPS不仅是生物除磷过程的中转站,而且参与了生物聚磷过程.     

Research on polyhydroxyalkanoates and glycogen transformations: Key aspects to biologic nitrogen and phosphorus removal in low dissolved oxygen systems

In this paper, a study was conducted on the effect of polyhydroxyalkanoates (PHA) and glycogen transformations on biologic nitrogen and phosphorus removal in low dissolved oxygen (DO) systems. Two laboratory-scale sequencing batch reactors (SBR1 and SBR2) were operating with anaerobic/aerobic (low DO, 0.15–0.45 mg·L⁻¹) configurations, which cultured a propionic to acetic acid ratio (molar carbon ratio) of 1.0 and 2.0, respectively. Fewer poly-3-hydroxybutyrate (PHB), total PHA, and glycogen transformations were observed with the increase of propionic/acetic acid, along with more poly-3-hydroxyvalerate (PHV) and poly-3-hydroxy-2-methyvalerate (PH2MV) shifts. The total nitrogen (TN) removal efficiency was 68% and 82% in SBR1 and SBR2, respectively. In the two SBRs, the soluble ortho-phosphate (SOP) removal efficiency was 94% and 99%, and the average sludge polyphosphate (poly-P) content (g·g-MLVSS⁻¹) was 8.3% and 10.2%, respectively. Thus, the propionic to acetic acid ratio of the influent greatly influenced the PHA form and quantity, glycogen transformation, and poly-P contained in activated sludge and further determined TN and SOP removal efficiency. Moreover, significant correlations between the SOP removal rate and the (PHV + PH2MV)/PHA ratio were observed (R ²>0.99). Accordingly, PHA and glycogen transformations should be taken into account as key components for optimizing anaerobic/aerobic (low DO) biologic nitrogen and phosphorus removal systems.     

丙酸/乙酸比例对生物除磷系统的影响研究

在5个SBR反应器(1#～5#)中,用乙酸、丙酸/乙酸(以C计)比例为0.5、2、10和丙酸的合成废水,通过厌氧/好氧工艺驯化活性污泥使之富集聚磷菌,研究了丙酸/乙酸比例对强化生物除磷系统(EBPR)中COD降解、溶解性正磷酸盐(SOP)的释放、吸收及其去除率的影响。结果表明,PAO对COD的降解符合一级动力学过程,PAO对丙酸的利用速率较乙酸快,因此,增加丙酸/乙酸比例有助于EBPR系统的稳定性。随丙酸/乙酸比例增加,SOP的释放与吸收量减少,SOP的代谢速率降低,但SOP的去除率依次为88.2%,89.8%,92.2%,94.2%和95.2%,去除率增加了7%,因此增加丙酸/乙酸比例有助于提高EBPR系统除磷效率。     

除磷颗粒诱导的同步短程硝化反硝化除磷颗粒污泥工艺

以低C/N比生活污水为研究对象,接种成熟除磷颗粒污泥,通过联合调控好氧时间及曝气强度成功将其诱导成具有同步短程硝化反硝化除磷功能的颗粒污泥,并分析了此过程中系统脱氮除磷特性变化.结果表明,好氧段曝气强度为5L·(h·L)^-1,在较短曝气时间下(140 min)可实现AOB的富集,但同步硝化反硝化能力难以提高;降低曝气强度为3. 5L·(h·L)^-1,延长曝气时间(200 min),好氧段氮损增加.根据pH及DO曲线进一步优化曝气时长抑制NO₂^-向NO₃^-转化,优化后系统出水TP <0. 5 mg·L^-1和TN <15 mg·L^-1,可实现氮磷的同步去除.在系统功能由单纯的除磷向同步脱氮除磷转化的过程中,释磷量下降,PAOs在内碳源储存过程中的贡献比例有所下降,但仍占主体地位(60%).批次实验表明,颗粒中可利用NO₂^-为电子受体的DPAOs占绝大部分达52....     

有机碳源对启动及运行CANON颗粒污泥工艺的影响

采用两组平行的SBR反应器R1和R2,通过分别添加有机碳源和不添加有机碳源对比实验,研究有机碳源对全程自养脱氮(CANON)颗粒污泥工艺启动的影响,通过改变有机碳源浓度对比实验,研究有机碳源对CANON工艺运行的影响.结果表明,50 mg·L-1有机物的存在可以加快CANON颗粒污泥工艺的启动,R1和R2分别于23 d和32 d成功启动CANON颗粒污泥工艺.适量有机物(不超过150 mg·L-1)的存在会通过增强亚硝化菌(AOB)及厌氧氨氧化菌(An AOB)的活性和促进反硝化作用来提高氨氮及总氮去除率,R1和R2的最高氨氮和总氮去除率分别为92%、88%和89%、80%.进一步实验表明,过量的有机碳源(超过200 mg·L-1)会抑制AOB及An AOB的活性,从而降低氨氮及总氮去除率,但是会促进反硝化反应并提高其对系统脱氮的贡献率.     

Influence of carbon source and temperature on the denitrifying phosphorus removal process

To supply the valuable operating parameters for the popular usage of the new denitrifying phosphors removal process, it is essential to study the dominant biochemical reactions and the characteristics of denitrifying phosphorus removing bacteria (DPB). Thus, parallel batch experiments using DPB sludge were carried out to assess the effect of substrates (sewage, HAc, and endogenous carbon source) on denitrifying dephosphorus removal efficiency in this study. The results showed that the initial specific phosphorus release rate increased with the high concentration of the short-chain volatile fatty acids ratio in the influent, and sufficient phosphorus was released by DPB. This improved the subsequent denitrification and phosphorus uptake efficiency. The specific endogenous denitrification mainly relies on the internal carbon source (PHB) stored by poly-P bacteria. Denitrifying phosphorus removing bacteria were very hungry when the internal PHB was consumed. Consequently, the specific endogenous denitrification rate was low and the phosphorus uptake did not happen. On the other hand, in the experiment, the denitrifying phosphorus removal performance under two temperature conditions (8–10°C and 25–26°C) was also investigated and analyzed. It was found that the lower temperature decreased the specific phosphorus release and uptake rate, but did not inhibit the denitrifying phosphorus removal completely. Therefore, the negative influence of the low temperature on the overall phosphorus removal was not significant. Translated from Acta Scientiae Circumstantiae, 2006, 26 (2): 186–192 [译自: 环境科学学报]     

好氧颗粒污泥处理制糖工业废水厌氧出水的除磷特性研究

制糖工业废水经厌氧生物处理后,COD大幅下降,但是出水中N、P含量仍然较高,严重破坏水体生态平衡.利用好氧颗粒污泥对制糖工业废水的厌氧出水进行脱氮除磷处理,讨论了其除磷过程.经复合底物(乙酸盐、丙酸盐、丁酸盐)培养的好氧颗粒污泥直径1.7 mm,SVI为38.43 mL.g-1,TP去除率达90.9%,出水磷含量仅为1.3 mg.L-1,单位COD释磷率为0.571,厌氧条件下磷的释放速率达到5.73 mg.(g.h)-1,好氧颗粒污泥表现出较好的沉淀性能和较高的除磷活性.由于底物中丙酸盐、丁酸盐含量增加,使得聚磷菌在反硝化过程中NO3--N的利用率增加,即消耗单位质量的NO3--N可以吸收更多的磷.好氧颗粒污泥及其胞外聚合物中P元素的含量与其中Mg、Ca、Fe元素的含量表现出很高的相关性,胞外聚合物对P的吸附使得体系除磷能力进一步增强.通过对污泥反硝化除磷的研究发现,反硝化聚磷菌占总聚磷菌的61.9%,其吸磷量与消耗硝酸盐的比值[m(P)/m(NO3--N)]为1.14.     

Zn2+对SBR单级好氧模式生物除磷性能的影响

以合成废水为研究对象,以丙酸盐为单一外加碳源,通过比较进水中不同Zn2+浓度(0,1,5,10,20mg/L)下单级好氧模式下序批式反应器(SBR)的除磷效果,考察进水Zn2+浓度对单级好氧SBR生物除磷性能的影响,并通过分析各反应器中典型周期内磷及微生物体内储能物质的变化,探究Zn2+对单级好氧SBR生物除磷性能的影响机理.当进水Zn2+浓度为0和1mg/L时,除磷率分别高达96.84%和97.90%.当进水Zn2+浓度为5,10,20mg/L时,系统除磷率分别为89.32%,76.43%和57.29%,说明较高浓度Zn2+对单级好氧SBR生物除磷有抑制作用.结果表明,较高浓度Zn2+可抑制COD的降解,微生物体内聚羟基脂肪酸酯好氧合成及磷酸盐激酶活性,并促进GAOs的代谢,使聚磷合成和水解量减少,从而降低系统的除磷性能.     

聚烃基烷酸转化对强化生物除磷影响研究

通过丙酸和乙酸C-mol比为0.5和2的合成废水驯化微生物的SBR反应器(SBR1和SBR2)批式实验,研究了强化生物除磷系统中聚烃基丁酸(PHB)和聚烃基戊酸(PHV)的转化对磷吸收/释放及去除率的影响.结果显示,磷的释放/吸收和去除率与PHB和PHV的转化有很好的相关性(R²>0.90).回归系数表明,特定废水驯化的污泥,磷的吸收和释放主要受PHB转化的影响,但磷的去除率却主要依赖于PHV的合成与降解;对于不同比例丙酸/乙酸废水驯化污泥,SBR2比SBR1污泥的PHB合成和降解能力增强,PHV合成和降解能力减小,生物除磷效果平均增加16.69%.因此,进水丙酸/乙酸比例及驯化影响聚磷微生物的PHB/PHV转化量,进而影响对磷的吸收/释放和除磷效果,PHB与PHV的转化量应作为生物除磷系统的关键调控因素考虑.