Performance and microbial community analysis of bioaugmented activated sludge for nitrogen-containing organic pollutants removal

Nitrogen-containing organic pollutants (quinoline, pyridine and indole) are widely distributed in coking wastewater, and bioaugmentation with specific microorganisms may enhance the removal of these recalcitrant pollutants. The bioaugmented system (group B) was constructed through inoculation of two aromatics-degrading bacteria, Comamonas sp. Z1 (quinoline degrader) and Acinetobacter sp. JW (indole degrader), into the activated sludge for treatment of quinoline, indole and pyridine, and the non-bioaugmented activated sludge was used as the control (group C). Both groups maintained high efficiencies (> 94%) for removal of nitrogen-containing organic pollutants and chemical oxygen demand (COD) during the long-term operation, and group B was highly effective at the starting period and the operation stage fed with raw wastewater. High-throughput sequencing analysis indicated that nitrogen-containing organic pollutants could shape the microbial community structure, and communities of bioaugmented group B were clearly separated from those of non-bioaugmented group C as observed in non-metric multidimensional scaling (NMDS) plot. Although the inoculants did not remain their dominance in group B, bioaugmentation could induce the formation of effective microbial community, and the indigenous microbes might play the key role in removal of nitrogen-containing organic pollutants, including Dokdonella, Comamonas and Pseudoxanthomonas. Phylogenetic Investigation of Communities by Reconstruction of Unobserved States (PICRUSt) analysis suggested that bioaugmentation could facilitate the enrichment of functional genes related to xenobiotics biodegradation and metabolism, probably leading to the improved performance in group B. This study indicated that bioaugmentation could promote the removal of nitrogen-containing organic pollutants, which should be an effective strategy for wastewater treatment.     

利用T-RFLP解析生物强化去除吡啶过程中微生物种群动态变化

在接种活性污泥处理含吡啶废水的序批式反应器中,引入吡啶降解菌Paracoccus sp.KT-5构成生物强化反应器,研究了对吡啶的生物强化去除特性及效果,利用末端限制性片段长度多态性(T-RFLP)手段解析了微生物种群结构的动态变化.结果表明,投加高效降解菌株KT-5可以加速反应器的启动,但随着反应器的运行,当吡啶初始浓度增加至195.6 mg·L^-1以后,生物强化反应器对吡啶降解的促进作用已不再明显;当吡啶初始浓度在293.4~586.8 mg·L^-1变化时,起初强化反应器对吡啶的去除速率出现了波动,尽管随后逐渐恢复,但仍然没有表现出明显的强化作用.T-RFLP的分析结果表明,当吡啶初始浓度达到978 mg·L^-1以后,生物强化反应器中已检测不到KT-5,表明生物强化作用的消失可能是因为引入的高效降解菌株KT-5的流失造成的.     

基因工程菌生物强化MBR工艺处理阿特拉津试验研究

以生活污水为共基质,考察了基因工程菌在MBR和活性污泥反应器中对阿特拉津的生物强化处理效果,以及生物强化处理对污泥性状的影响.结果表明,基因工程菌在MBR中对阿特拉津具有很好的生物强化处理效果,阿特拉津平均出水浓度为0.84 mg/L,平均去除率为95%,最大去除负荷可以达到70 mg/(L·d).生物强化的MBR对生活污水中COD的平均去除率为71%,COD平均出水浓度65 mg/L,COD容积负荷增加对COD去除效果有一定影响;对生活污水中的氨氮具有很好的去除效果,氨氮平均出水浓度为1.1 mg/L,平均去除率为97%,最大氨氮去除负荷为143 mg/(L·d).与普通MBR污泥相比,生物强化MBR污泥的硝化活性和亚硝化活性略高,碳氧化活性略低,因此表现出氨氮处理效果很好,COD处理效果略差.阿特拉津的存在会对污泥性状产生影响,可能是造成污泥碳氧化活性低的原因.     

Enhanced atrazine removal using membrane bioreactor bioaugmented with genetically engineered microorganism

Bioaugmentation with genetically engineered microorganisms (GEMs) in a membrane bioreactor (MBR) for enhanced removal of recalcitrant pollutants was explored. An atrazine-degrading genetically engineered microorganism (GEM) with green fluorescent protein was inoculated into an MBR and the effects of such a bioaugmentation strategy on atrazine removal were investigated. The results show that atrazine removal was improved greatly in the bioaugmented MBR compared with a control system. After a start-up period of 6 days, average 94.7% of atrazine was removed in bioaugmented MBR when atrazine concentration of influent was 14.5 mg/L. The volumetric removal rates increased linearly followed by atrazine loading increase and the maximum was 65.5 mg/(L·d). No negative effects were found on COD removal although carbon oxidation activity of bioaugmented sludge was lower than that of common sludge. After inoculation, adsorption to sludge flocs was favorable for GEM survival. The GEM population size initially decreased shortly and then was kept constant at about 10⁴–10⁵ CFU/mL. Predation of micro-organisms played an important role in the decay of the GEM population. GEM leakage from MBR was less than 10² CFU/mL initially and was then undetectable. In contrast, in a conventionally activated sludge bioreactor (CAS), sludge bulking occurred possibly due to atrazine exposure, resulting in bioaugmentation failure and serious GEM leakage. So MBR was superior to CAS in atrazine bioaugmentation treatment using GEM.     

生物强化去除吡啶的特性及微生物种群动态变化分析

在接种活性污泥的序批式反应器中投加固定化吡啶降解菌Paracoccus sp.KT-5,强化吡啶的生物降解,并与未投加固定化微生物的反应器进行对照,通过末端限制性片段长度多态性（T-RFLP）分析手段,探讨了运行过程中生物强化与未强化反应器中微生物群落结构的动态变化,并探讨了生物强化反应器的去除效果.结果表明,投加固定化吡啶降解菌可以加速反应器的启动.当吡啶初始浓度为195.6～586.8 mg.L-1,随着反应器的运行,投加固定化菌株的强化作用并不明显;但当吡啶初始浓度为782.4～2 934 mg.L-1,投加固定化菌株显示出优势.T-RFLP分析结果表明,投加的固定化菌株KT-5作为优势菌始终存在于反应器的固定化生物相和悬浮生物相中.     

生物强化MBR-AF工艺短程硝化反硝化研究

本试验将短程硝化功能菌和反硝化功能菌分别接种至膜生物反应器(MBR)和上流式厌氧生物滤池(AF)中,构建了生物强化的MBR-AF短程硝化反硝化工艺,并以活性污泥作空白对照,考察该工艺对高氨氮废水的短程脱氮性能.结果表明,强化体系MBR的启动期短,仅需30d,而活性污泥体系MBR的启动期长达100d;强化体系MBR亚硝酸氮积累率始终维持在95%以上;在30℃下,随着运行时间的延长,强化体系MBR-AF工艺总氮去除率不断升高,最高达90%以上,比活性污泥体系高20%;强化体系MBR膜污染程度轻,膜的使用寿命长.说明功能菌强化在高效亚硝酸氮积累和氨氮转化方面起关键作用,可作为实现短程硝化反硝化的有效手段.     

接合子细胞强化对生物反应器降解2，4-D效应研究

研究了接合性质粒pJP4在2株纯菌E．coliDH5α、Alcaligenes sp．及好氧颗粒污泥混合菌群中的水平转移情况,并以获得pJP4质粒的接合子Alcaligenes sp．：：pJP4为强化菌种,考察了接合子细胞强化对好氧颗粒反应器和生物膜反应器中难降解有机物2,4-二氯苯氧乙酸（2,4-D）的去除效应．结...     

酚降解菌株的分离、鉴定和在含酚废水生物处理中的应用

从石油化工厂的活性污泥和废水混合物中分离到10个降解酚的细菌菌株,经16S rRNA基因序列分析,5个菌株(PD1、PD2、PD6、PD7和PD39)被鉴定为假单胞菌(Pseudomonas sp.),4个菌株(PD4、PD5、PD8和PD9)为不动杆菌(Acinetobacter sp.),1个菌株(PD3)为丛毛单胞菌(Comamonas sp.).对假单胞菌(Pseudomonas sp.)PD39菌株的酚降解特性、最适生长条件、降解底物的范围、儿茶酚1,2-双加氧酶(C12O)和儿茶酚2,3-双加氧酶(C23O)活性、含酚废水的生物处理等,进行了详细研究.结果表明,PD39菌株的最适生长和降解条件是,培养基的起始pH为7.0,培养温度为30℃,酚浓度为800 mg/L,对酚的最大降解浓度为1 200 mg/L.酚浓度为637 mg/L的工业废水经72 h处理酚的去除率达到99.96%.PD39菌株有很好的应用潜力来作含酚废水活性污泥处理系统的强化菌株.     

Shifts in microbial community structure and diversity in a MBR combined with worm reactors treating synthetic wastewater

The chemical oxygen demand (COD) and NH₃-N removal, membrane fouling, sludge characteristics and microbial community structure in a membrane bioreactor (MBR) coupled with worm reactors (SSBWR) were evaluated for 210 days. The obtained results were compared to those from a conventional MBR (C-MBR) operated in parallel. The results indicated that the combined MBR (S-MBR) achieved higher COD and NH₃-N removal efficiency, slower increase in membrane fouling, better sludge settleability and higher activities of the related enzymes in the activated sludge. Denaturing gradient gel electrophoresis was used to analyze the microbial community structures in the C-MBR and the S-MBR. The microbial community structure in the S-MBR was more diverse than that in the C-MBR. Additionally, the slow-growing microbes such as Saprospiraceae, Actinomyces, Frankia, Clostridium, Comamonas, Pseudomonas, Dechloromonas and Flavobacterium were enriched in the S-MBR, further accounting for the sludge reduction, membrane fouling alleviation and wastewater treatment.     

Bioaugmentation and quorum sensing disruption as solutions to increase nitrate removal in sequencing batch reactors treating nitrate-rich wastewater

Bioaugmentation of denitrifying bacteria can serve as a promising technique to improve nutrient removal during wastewater treatment. While denitrification inhibition by bacterial quorum sensing (QS) in Pseudomonas aeruginosa has been indicated, the application of bacterial QS disruption to improve nitrate removal from wastewater has not been investigated. In this study, the effect of bioaugmentation of P. aeruginosa SD-1 on nitrate removal in sequencing batch reactors that treat nitrate rich wastewater was assessed. Additionally, the potential of a quorum sensing inhibitor (QSI) to improve denitrification following bacterial bioaugmentation was evaluated. Curcumin, a natural plant extract, was used as a QSI. The chemical oxygen demand (COD) and initial nitrate concentration of the influent were 700±20 mg/L and 200±10 mg/L respectively, and their respective concentrations in the effluent were 56.9±3.2 mg/L and 9.0±3.2 mg/L. Thus, the results revealed that bioaugmentation of P. aeruginosa SD-1 resulted in an increased nitrate removal to 82%±1%. Further, nitrate was almost completely removed following the addition of the QSI, and activities of nitrate reductase and nitrite reductase increased by 88%±2% and 74%±2% respectively. The nitrogen mass balance indicated that aerobic denitrification was employed as the main pathway for nitrogen removal in the reactors. The results imply that bioaugmentation and modulation of QS in denitrifying bacteria, through the use of a QSI, can enhance nitrate removal during wastewater treatment.     

高效水解酸化UASB活性污泥的菌群结构分析

采用454高通量测序技术对低能耗、低污泥产量且具有脱氮效能的印染废水处理工艺中UASB污泥的微生物菌群结构进行了分析.结果表明,UASB内污泥的微生物菌种呈多样性分布且优势菌群突出,通过菌群鉴定发现,脱硫橄榄样菌属(Desulfobacula)、Levilinea、长绳菌属(Longilinea)、Candidatus Tammella、Paludibacter、索氏菌属(Thauera)、Tepidimicrobium、杆状脱硫菌属(Desulforhabdus)、类芽孢杆菌属(Paenibacillus)、梭菌属(Clostridium)是主要的优势菌属.其中,梭菌属是起到产酸和污泥减量作用的主要菌种.另外,具有脱氮效能的原因可能是由于发生了硫酸盐型厌氧氨氧化作用.通过Shannon、Chao、Simpson、Shannon指数的计算,发现该UASB中微生物较其他废水处理系统,群落结构拥有较高的多样性和丰度,有利于稳定产酸.     

复合菌群生物强化削减NPEO内分泌干扰毒性的试验研究

印染废水中的壬基酚聚氧乙烯醚(nonylphnol polyethylene ether, NPEO)在生化处理过程中会产生雌激素活性更强的壬基酚(NP)等中间产物,导致处理后印染废水内分泌干扰毒性升高.为探索以NPEO和NP为降解靶点进行菌群生物强化脱毒的可行性,分别以NPEO和NP驯化富集印染活性污泥,并将得到的降解菌群以单独和组合投加方式进行生物强化试验,考察强化控毒效果.结果表明：(1)NPEO降解菌群(NPEB)和NP降解菌群(NPB)中的优势菌均为Proteobacteria,二者对10 mg/L NPEO和NP的48 h去除率均高于98%.(2)单独或组合投加5 mg/L的NPEB和NPB至混合液悬浮固体浓度(MLSS)为500 mg/L的活性污泥体系,均能显著提升活性污泥对不同浓度(10和1 mg/L) NPEO的降解性能,大幅缩短NPEO降解过程中雌激素活性的变化周期,并使体系的雌激素活性维持在较低水平.(3)当降解体系中加入1 000 mg/L葡萄糖作为额外碳源时,NPB的强化性能被完全抑制,而NPEB在降解性能受抑的情况下仍能增强活性污泥的NPEO降解速率并缩短控...     

不同生物反应器中基因工程菌生物强化处理阿特拉津研究

在膜-生物反应器(MBR)和复合生物反应器中,考察基因工程菌生物强化处理阿特拉津去除效果,并对基因工程菌浓度和降解基因atzA基因丰度变化进行检测.结果表明,阿特拉津对COD和氨氮的生物去除活性具有一定的抑制作用;基因工程菌生物强化后,COD及氨氮的去除效率得到恢复.MBR对COD和氨氮的去除效果优于复合生物反应器.基...     

MBR活性污泥培养驯化过程中生物多样性研究

以MBR反应器启动调试阶段的活性污泥为主要研究对象,系统考察了传统活性污泥法(Conventional activated sludge,CAS)污泥接种至MBR反应器内污泥培养驯化过程中生物多样性情况及微生物群落结构的演变规律.同时,在传统污水污泥检测指标的基础上,对各阶段污泥中总细菌基因组DNA进行提取,应用PCR-DGGE分子生物学技术获得了相应的凝胶电泳图谱并进一步分析了菌群间的相似性.结果表明,以CAS污泥为接种污泥在MBR反应器内培养驯化过程中微生物的多样性变化突出,细菌群落结构演替明显,不同阶段菌群间的相似性说明了各阶段菌群的演变关系:污泥培养驯化是一个逐步有序的过程,微生物随反应器内不同时期及环境的变化而调整,逐渐演变成适应MBR工艺的群落结构.     

利用PCR-DGGE研究膜生物反应器中微生物的群落结构

使用聚合酶链式反应-变性梯度凝胶电泳技术(PCR-DGGE)考查了天津某再生水处理厂膜生物反应器(Membrane Bioreactor,简称MBR)培养驯化直至正常运行全过程中总细菌群落结构的演替情况.结果表明,在MBR环境中,接种的传统活性污泥中的微生物群落在几天内发生了较大的变化,在进污水驯化时,微生物群落也遭受了冲击,最后经过培养驯化趋于稳定,一些菌种逐渐成长为顶级优势微生物,在反应器内占据主导地位.最终该反应器逐渐形成了自己独有的微生物群落生态系统.另外,对该微生物群落的部分优势总细菌进行了克隆测序和系统发育树分析,通过鉴定获得的10条总细菌的16S rDNA序列,它们分别与气单胞菌属、假单胞菌、亚硝酸菌属、丛毛单胞菌属和杆菌的同源性在97%以上,这些优势微生物在MBR反应器去除有机物的过程中起到了关键的作用.     

2株好氧反硝化菌的筛选及其强化贫营养生物膜脱氮效果

经富集培养、BTB培养基初筛与反硝化能力测定,从城市污水处理厂活性污泥中筛选得到2株好氧反硝化细菌.通过16S rDNA同源性分析对2株菌进行鉴定;并将这2菌株接种到贫营养生物膜体系中以探究它们对系统总氮的去除能力的强化.结果表明,这2株菌分别属于Pseudomonas aeruginosa和Pseudomonas putida,2株好氧反硝化菌单独存在时,对模拟废水的TN去除率分别达78%和82%;2株细菌强化后的生物膜系统对TN去除率达68%和64%,较非强化对照系统分别提高47%和43%,且NH4+-N去除率均接近100%.说明这2株好氧反硝化细菌具有较强的反硝化能力,并能够有效强化生物膜在贫营养条件下的反硝化能力,并且不会抑制生物膜硝化能力,可实现生物膜系统同步硝化反硝化.     

Analysis of bacterial community in bulking sludge using culture-dependent and -independent approaches

Combination of 16S rRNA gene clone library and cultivation for assessing of bacterial diversity of the microflora in bulking sludge was evaluated in the study.     

气升循环分体式膜生物反应器再生回用厕所污水的研究

研究了分体浸没式膜生物反应器及其应用于厕所污水的再生回用.试验结果表明,在人水浓度COD440～970mg/L、BOD₅ 307～612mg/L、NH₃-N59～111mg/L的情况下,出水水质稳定,平均COD＜47mg/L、BOD₅＜8.5mg/L、NH₃-N＜20mg/L,处理水质达到建设部颁布的生活杂用水回用标准.研究了动力学参数污泥负荷、容积负荷和HRT对出水COD的影响,确定本试验较佳的污泥负荷和容积负荷分别为0.1kg/(kg·d)和1kg/(m³·d),HRT为7～8h.本文对污泥浓度、温度和产水时间与间歇时间比对膜通量的影响进行了测定与探讨.     

Influence of microbial community structure of seed sludge on the properties of aerobic nitrifying granules

In order to evaluate the influence of microbial community structure of seed sludge on the properties of aerobic nitrifying granules, these granules were cultivated with different seed sludge, and the variation of microbial community and dominant bacterial groups that impact the nitrogen removal efficiency of the aerobic nitrifying granules were analyzed and identified using 16s rDNA sequence and denaturing gradient gel electrophoresis (DGGE) profiles. The results presented here demonstrated that the influence of the community structure of seed sludge on the properties of aerobic nitrifying granules was remarkable, and the granules cultivated by activated sludge from a beer wastewater treatment plant showed better performance, with a stable sludge volume index (SVI) value of 20 mL/g, high extracellular polymeric substance (EPS) content of 183.3 mg/L, high NH₄⁺-N removal rate of 89.42% and abundant microbial population with 10 dominant bacterial groups. This indicated that activated sludge with abundant communities is suitable for use as seed sludge in culturing aerobic nitrifying granules.     

基因工程菌生物强化处理系统微生物群落分析

在膜-生物反应器(MBR)和传统活性污泥反应器(CAS)中,考察了基因程菌生物强化对阿特拉津的去除效果,并通过PCRDGGE分析了反应器不同运行阶段污泥微生物群落的变化.结果表明,基因工程菌生物强化实现了阿特拉津的高效生物去除,MBR和CAS阿特拉津平均去除率分别达到88.6％和85.3％.阿特拉津生物强化去除有助于反...