Effect of nitrite on hydrolysis-acidification, biogas production and microbial community in semi-continuous two-phase anaerobic digestion of sewage sludge

Previous study found that the pre-treatment of sewage sludge with nitrite improves the biogas production during the mono/two-phase anaerobic digestion (AD) using batch biochemical methane potential tests. In this study, the effects of nitrite on hydrolysis-acidification, biogas production, volatile solids destruction and microbial composition in semi-continuous two-phase AD of sewage sludge were investigated. The addition of nitrite promotes sludge organic matter solubilization (+484%) and VFAs production (+98.9%), and causes an increase in the VS degradation rate during the AD process (+8.7%). The comparison of biogas production from the acidogenic and methanogenic reactors with or without the addition of nitrite implies that the nitrite has no significant effect on the overall biogas production of two-phase sludge AD process. High-throughput sequencing analysis shows that the microbial communities of bacteria and archaea in two-phase AD reactors significantly changes after the addition of nitrite. Vulcanibacillus (bacteria) and Candidatus Methanofastidiosum (archaea) become the dominant genera in the acidogenic and methanogenic reactors with the nitrite respectively. These findings provide new insights about using nitrite to promote the organic matter degradation of sewage sludge in a semi-continuous two-phase AD system.     

Microbial characteristics in anaerobic membrane bioreactor treating domestic sewage: Effects of HRT and process performance

Two anaerobic membrane bioreactors (AnMBRs) equipped with different membrane pore size (0.4 or 0.05μm) were operated at 25℃ and fed with domestic wastewater.The hydraulic retention time (HRT) of the reactors was shortened.The microbial communities of the two AnMBRs were investigated by 16S rRNA gene amplicon sequencing to see the effects of HRT.The predominant Archaea was an aceticlastic methanogen Methanosaeta.The composition of hydrogenotrophic methanogens changed with the HRTs:the population ...     

耐盐碱水稻土壤产甲烷菌群落特征及产甲烷途径

产甲烷菌在土壤碳循环过程中发挥着关键性作用,而种植耐盐碱水稻土壤中产甲烷菌群落特征及环境影响因素尚不明确.本研究采用实时荧光定量PCR和Illumina高通量测序技术,对青岛市稻作改良智慧农业产业示范基地中耐盐碱水稻（YC1703）、普通水稻（"临稻10号"）及荒地土壤中产甲烷菌的丰度特征、群落组成及与环境因子的关系进行了对比分析.结果表明,种植"临稻10号"土壤中的产甲烷菌丰度和群落丰富度显著高于种植YC1703的土壤,种植YC1703土壤中产甲烷菌具有较高多样性.产甲烷菌群落受水稻品种、水稻生育期和环境因子的综合影响.两种水稻土壤中H₂/CO₂还原型产甲烷菌占优势,由此推测这些土壤中生成甲烷的主导途径均为H₂/CO₂还原途径.     

PCR-DGGE研究青海农村户用沼气池微生物群落结构

采用变性梯度凝胶电泳(DGGE)分析技术,对青海农村户用沼气池内细菌及古菌群落结构进行分析.结果表明,沼气池含有丰富的细菌及古菌类群,且泥样间的细菌和古菌的群落结构存在差异.4个户用沼气池发酵系统中,细菌分属于5个门,优势类群为拟杆菌门(Bacteroidetes)、厚壁菌门(Firmicutes)和变形菌门(Proteobacteria).在属分类水平上,属于24个类群,最优势类群为理研菌科佩特里单胞菌属(Petrimonas)、假单胞菌属(Pseudomona)、泰氏菌属(Tissierella)和梭菌属(Clostridium).同时,沼气池发酵系统中古菌主要包括热丝菌属(Thermofilum)、甲烷短杆菌属(Methanobrevibacter)、甲烷囊菌属(Methanoculleus)、产甲烷菌属(Methanogenium),其中,Methanobrevibacter和Methanogenium是沼气池内最优势的产甲烷菌.说明沼气池的产甲烷途径主要是氢营养代谢类型,水解、酸化过程主要由来自动物消化系统内的细菌完成.     

发酵温度对青海农用沼气池微生物群落的影响

为了研究发酵温度对青海农用沼气池微生物群落结构的影响,以沼气池全年6个温度时期的24个泥样为研究对象,采用变性梯度凝胶电泳分析样品中细菌与古菌的群落结构及其变化差异.结果显示：发酵温度的波动变化对微生物群落结构、多样性及沼气池产气量影响显著.细菌在温度最高（九月中旬）和最低（三月中旬）的2个采样时期多样性最高,古菌仅在温度最高时期多样性最高.在细菌类群中,厚壁菌门Firmicutes、拟杆菌门Bacteroidetes和变形菌门Proteobacteria是所有样品中丰度最高的类群（三者总和>72.43%）,是发酵系统中重要的功能细菌类群.细菌属分类水平上,理研菌科佩特里单胞菌属Petrimonas、梭菌属Clostridium、泰氏菌属Tissierella和假单胞菌属Pseudomona是所有样品的优势类群,总丰度比例较为恒定（约32.40%）.在古菌类群中,甲烷微菌目是最优势类群,丰度为40.62%~64.74%.产甲烷菌属Methanogenium是所有样品中主要的产气功能种群,丰度波动范围为32.62%~55.74%,随全年温度的变化而波动,此与沼气的产气规律相一致.     

乙酸驯化对厌氧污泥微生物群落结构及发酵特性的影响

采用乙酸对厌氧污泥进行逐步驯化,以富集乙酸营养型产甲烷菌群,解决厌氧发酵过程中的酸抑制问题.对驯化前后污泥中的微生物群落结构及其在高酸浓度和低p H值条件下的发酵特性进行了研究.结果表明:驯化后污泥中乙酸营养型产甲烷菌中的甲烷八叠球菌属(Methanosarcina)得到了明显富集,其相对丰度由原始的4.2%提高到58.1%,成为耐酸污泥中的主导优势古菌群;氢营养型产甲烷菌属的丰度则都有不同程度的下降.污泥中产甲烷菌群由氢营养型为主导转为乙酸营养型和氢营养型共同主导.驯化前后污泥中细菌的优势菌门均为主要降解纤维素和半纤维素的厚壁菌门(Firmicutes)和降解蛋白质的拟杆菌门(Bacteroidetes),其中驯化后Firmicutes的丰度由48.8%提高到61.7%,而Bacteroidetes的丰度则由30.1%降低至16.9%.驯化后的污泥对高VFA浓度和低pH值的耐受性均有较大程度的提高,其在VFA浓度为7500 mg·L~(-1)及pH 6.0条件下仍可以快速产气.     

基于固液分离预处理的餐厨垃圾厌氧发酵

研究不同预处理方法所获餐厨垃圾浆料的半连续式厌氧发酵效果,评价该类餐厨垃圾的产甲烷资源化潜能.结果表明,固液油三相分离预处理后的液相餐厨垃圾作为进料较固液混合相餐厨垃圾在厌氧发酵时具有更高的VS去除率,实际产甲烷潜能和甲烷转化率,分别为91.2%, 531.5mLCH₄/gVS和54.3%,表明液相餐厨垃圾半连续中温湿式厌氧发酵具有良好的减量化和资源化效能.微生物群落分析表明,不同预处理方式影响了餐厨垃圾厌氧发酵系统的微生物群落演替,氨氮浓度是导致古菌群落转移和丰度变化的关键因素.液相餐厨垃圾厌氧发酵时是以氢营养型产甲烷菌Methanoculleus为主,而在固液混合相餐厨垃圾厌氧发酵时则以耐高浓度氨氮的Methanosarcina多功能产甲烷菌为主.     

PCR-DGGE技术在城市污水化学生物絮凝处理中的特点

通过PCR-DGGE等分子生物学技术可以不经过常规培养直接从活性污泥和生物膜样品中提取DNA,对16Sr DNA V3区进行PCR扩增,结合DGGE(变性梯度凝胶电泳),从而分析活性污泥与生物膜中微生物种群结构.研究证实,活性污泥培养前后微生物种群结构发生很大的改变.同时对2种污水处理工艺中微生物种群结构进行了对比研究,对同一反应器不同位置微生物分布以及不同工况下的微生物种群结构进行了初步探讨.测定了活性污泥中部分菌种的16S rDNA V3区片段序列,通过NCBI(美国国立生物技术信息中心)基因库比对,初步确定细菌的属.结果显示,PCR-DGGE结合测序技术是一种完全可行的快速进行环境学样品微生物研究的分析方法.     

Structure and activity of thermophilic methanogenic microbial communities exposed to quaternary ammonium sanitizer

Food processing facilities often use antimicrobial quaternary ammonium compound (QAC) sanitizers to maintain cleanliness. These QACs can end up in wastewaters used as feedstock for anaerobic digestion. The aim of this study was to measure the effect of QAC contamination on biogas production and structure of microbial communities in thermophilic digester sludge. Methane production and biogas quality data were analyzed in batch anaerobic digesters containing QAC at 0, 15, 50, 100 and 150 mg/L. Increasing sanitizer concentration in the bioreactors negatively impacted methane production rate and biogas quality. Microbial community composition data was obtained through 16S rRNA gene sequencing from the QAC-contaminated sludges. Sequencing data showed no significant restructuring of the bacterial communities. However, significant restructuring was observed within the archaeal communities as QAC concentration increased. Further studies to confirm these effects on a larger scale and with a longer retention time are necessary.     

温度对FBAR反应器的运行特性及古菌微生物群落影响

为探讨固定床厌氧反应器（FBAR）在不同温度下的运行特性及微生物群落变化,比较了高温（50℃）、中温（35℃）、低温（4℃）3个温度阶段反应器产甲烷特性及古菌群落变化.结果表明;绝对产气量由大至小依次为高（50℃）、中（35℃）、低温（4℃）,单位负荷产气量依次为中温（2.84L/OLR）,低温（2.5L/OLR）,高温（1.8L/OLR）;甲烷含量依次为低温（74.5%）、中温（63.5%）,高温（57.3%）,不同温度阶段对挥发性有机酸含量变化有一定的影响.克隆文库分析表明：不同温度条件下固定床厌氧反应器内部微生物群落的丰富性存在很大的差异.定量PCR分析表明：甲烷鬃毛菌是中温和高温反应器内的优势菌,低温4℃炭纤维载体和污泥中的优势菌都是甲烷微菌.从能耗、经济效益角度分析低温条件更适合沼气发酵,而主要是以嗜氢产甲烷菌代谢途径为主.     

驯化对餐厨垃圾厌氧消化系统微生物群落结构的影响

为研究驯化对餐厨垃圾厌氧消化系统微生物群落结构的影响,在单相完全搅拌式(CSTR)反应器内,以农村户用沼气池污泥为接种污泥,进行了餐厨垃圾中温厌氧消化.反应器在3 g·L-1·d-1(以VS计)的负荷下成功启动,并连续45 d维持性能稳定,表明驯化成功.期间采用454焦磷酸测序技术分析了驯化前后系统内的微生物群落结构.结果表明,微生物群落结构与底物密切相关,驯化后细菌及古菌群落都发生明显变化.从细菌群落看,与复杂有机物降解相关的菌类显著下降(如梭菌纲(Clostridia)和(vadin HA17),而易降解碳水化合物发酵菌(如Petrimonas)和脂肪降解菌(如Erysipelotrichia)显著增加.这与餐厨垃圾易降解有机物含量高,且富含淀粉和脂肪相关.丰富的易降解有机物还使得反应器内总挥发性脂肪酸(VFA)浓度((2203±174)mg·L-1)远高于种泥水平((222.0±0.3)mg·L-1),这导致了产甲烷菌由乙酸型的甲烷鬃菌属(Methanosaeta,占85.01%)绝对主导转向氢营养型的甲烷螺菌属(Methanospirillum,占35.35%)、甲烷囊菌属(Methanoculleus,占9.89%)与之(46.97%)共同主导的局面.然而,驯化后Methanosaeta在非最优条件下依然保持主导地位,可见接种污泥的群落组成对厌氧消化系统群落结构的塑造也具有重大影响.     

Use of gamma-irradiation pretreatment for enhancement of anaerobic digestibility of sewage sludge

The effects of γ-irradiation pretreatment on anaerobic digestibility of sewage sludge was investigated in this paper. Parameters like solid components, soluble components, and biogas production of anaerobic digestion experiment for sewage sludge were measured. The values of these parameters were compared before and after γ-irradiation pretreatment. Total solid (TS), volatile solid (VS), suspended solid (SS), volatile suspended solid (VSS), and average floc size of samples decreased after γ-irradiation treatment. Besides, floc size distribution of sewage sludge shifted from 80–100 μm to 0–40 μm after γ-irradiation treatment at the doses from 0 to 30 kGy, which indicated the disintegration of sewage sludge. Moreover, microbe cells of sewage sludge were ruptured by γ-irradiation treatment, which resulted in the release of cytoplasm and increase of soluble chemical oxygen demand (SCOD). Both sludge disintegration and microbe cells rupture enhanced the subsequent anaerobic digestion process, which was demonstrated by the increase of accumulated biogas production. Compared with digesters fed with none irradiated sludge, the accumulated biogas production increased 44, 98, and 178 mL for digesters fed sludge irradiated at 2.48, 6.51, and 11.24 kGy, respectively. The results indicated that γ-irradiation pretreatment could effectively enhance anaerobic digestibility of sewage sludge, and correspondingly, could accelerate hydrolysis process, shorten sludge retention time of sludge anaerobic digestion process. __________ Translated from Techniques and Equipment for Environmental Pollution Control, 2006, 7(8): 36–39 [译自: 环境污染治理技术与设备]     

Determination of the archaeal and bacterial communities in two-phase and single-stage anaerobic systems by 454 pyrosequencing

2-Phase anaerobic digestion (AD), where the acidogenic phase was operated at 2 day hydraulic retention time (HRT) and the methanogenic phase at 10 days HRT, had been evaluated to determine if it could provide higher organic reduction and methane production than the conventional single-stage AD (also operated at 12 days HRT). 454 pyrosequencing was performed to determine and compare the microbial communities. The acidogenic reactor of the 2-phase system yielded a unique bacterial community of the lowest richness and diversity, while bacterial profiles of the methanogenic reactor closely followed the single-stage reactor. All reactors were predominated by hydrogenotrophic methanogens, mainly Methanolinea. Unusually, the acidogenic reactor contributed up to 24% of total methane production in the 2-phase system. This could be explained by the presence of Methanosarcina and Methanobrevibacter, and their activities could also help regulate reactor alkalinity during high loading conditions through carbon dioxide production. The enrichment of hydrolytic and acidogenic Porphyromonadaceae, Prevotellaceae, Ruminococcaceae and unclassified Bacteroidetes in the acidogenic reactor would have contributed to the improved sludge volatile solids degradation, and ultimately the overall 2-phase system''s performance. Syntrophic acetogenic microorganisms were absent in the acidogenic reactor but present in the downstream methanogenic reactor, indicating the retention of various metabolic pathways also found in a single-stage system. The determination of key microorganisms further expands our understanding of the complex biological functions in AD process.     

Effect of a high strength chemical industry wastewater on microbial community dynamics and mesophilic methane generation

A high strength chemical industry wastewater was assessed for its impact on anaerobic microbial com- munity dynamics and consequently mesophilic methane generation. Cumulative methane production was 251 mL/g total chemical oxygen demand removed at standard temperature and pressure at the end of 30 days experimental period with a highest recorded methane percentage of 80.6% of total biogas volume. Volatile fatty acids （VFAs） analysis revealed that acetic acid was the major intermediate VFAs produced with propionic acid accumulating over the experimental period. Quantitative analysis of microbial communities in the test and control groups with quantitative real time polymerase chain reaction highlighted that in the test group, Eubacteria （96.3%） was dominant in comparison with methanogens （3.7%）. The latter were dominated by Methanomicrobiales and Methanobacteriales while in test groups increased over the experimental period, reaching a maximum on day 30. Denaturing gradient gel electrophoresis profile was performed, targeting the 16S rRNA gene of Eubacteria and Archaea, with the DNA samples extracted at 3 different time points from the test groups. A phylogenetic tree was constructed for the sequences using the neighborhood joining method. The analysis revealed that the presence of organisms resembling Syntrophomonadaceae could have contributed to increased production of acetic and propionic acid intermediates while decrease of organisms resembling Pelotomaculum sp. could have most likely contributed to accumulation of propionic acid. This study suggested that the degradation of organic components within the high strength industrial wastewater is closely linked with the activity of certain niche microbial communities within eubacteria and methanogens.     

UASB反应器中产甲烷菌对温度胁迫的响应

采用间歇培养方式探讨了升流式厌氧污泥床(UASB)反应器中不同营养类型产甲烷菌对温度胁迫的响应规律.研究结果表明,产甲烷螺旋菌(Methanospirillum)是该反应器的主要氢营养型产甲烷菌,而主要乙酸营养型产甲烷菌为产甲烷丝状菌(Methanosaeta).在35℃条件下,氢营养型和乙酸营养型产甲烷菌的累计甲烷产量分别为24.7,11.7mL,而最大产甲烷速率分别为0.74,0.18mL/h.当温度从35℃分别降低至30,25,20,15℃时,导致氢营养型产甲烷菌的累计产甲烷量分别减少了14.2%,34.0%,47.0%,57.5%,而乙酸营养型产甲烷菌的累计产甲烷量分别减少了5.1%,23.9%,45.3%,95.7%.由此可见,在20~30℃时氢营养型产甲烷菌对温度胁迫更加敏感,而在15℃以下时乙酸营养营养型产甲烷菌对温度胁迫更加敏感.     

Comparison of bacterial community structures in two systems of a sewage treatment plant using PCR-DGGE analysis

Bacterial community was investigated during sludge bulking period using combination of PCR amplification of 16sRNA genes with DGGE analysis.     

PCR-DGGE技术解析生物制氢反应器微生物多样性

为了揭示发酵法生物制氢反应器厌氧活性污泥的微生物种群多样性 ,从运行不同时期取厌氧活性污泥 ,通过细胞裂解直接提取活性污泥的基因组DNA .以细菌 16SrRNA基因通用引物F338GC/R5 34进行V3高变异区域PCR扩增 ,长约 200bp的PCR产物经变性梯度凝胶电泳 (DGGE)分离后 ,获得微生物群落的特征DNA指纹图谱 .研究表明 ,不同时期的厌氧活性污泥中存在共同种属和各自的特异种属 ,群落结构和优势种群数量具有时序动态性 ,微生物多样性呈现出协同变化的特征 .微生物多样性由强化到减弱 ,群落结构之间的相似性逐渐升高 ,演替速度由快速到缓慢 .优势种群经历了动态演替过程 ,最终形成特定种群构成的顶级群落 .     

Analysis of bacterial community structures in two sewage treatment plants with different sludge properties and treatment performance by nested PCR-DGGE method

The bacterial community structures in two sewage treatment plants with different processes and performance were investigated by denaturing gradient gel electrophoresis （DGGE） of nested polymerase chain reaction （nested PCR） amplified 16S rRNA gene fragments with group-specific primers. Samples of raw sewage and treated effluents were amplified using the whole-cell PCR method, and the activated sludge samples were amplified using the extracted genomic DNA before the PCR products were loaded on the same DGGE gel for bacterial community analysis. Ammonia-oxidizing bacterial and actinomycetic community analysis were also carried out to investigate the relationship between specific population structures and system or sludge performance. The two plants demonstrated a similarity in bacterial community structures of raw sewage and activated sludge, but they had different effluent populations. Many dominant bacterial populations of raw sewage did not appear in the activated sludge samples, suggesting that the dominant bacterial populations in raw sewage might not play an important role during wastewater treatment. Although the two plants had different sludge properties in terms of settleability and foam forming ability, they demonstrated similar actinomycetic community structures. For activated sludge with bad settling performance, the treated water presented a similar DGGE pattern with that of activated sludge, indicating the nonselective washout of bacteria from the system. The plant with better ammonium removal efficiency showed higher ammonia-oxidizing bacteria species richness. Analysis of sequencing results showed that the major populations in raw sewage were uncultured bacterium, while in activated sludge the predominant populations were beta proteobacteria.     

Impacts of produced water origin on bacterial community structures of activated sludge

The purpose of this study was to reveal how activated sludge communities respond to influent quality and indigenous communities by treating two produced waters from different origins in a batch reactor in succession. The community shift and compositions were investigated using Polymerase Chain Reaction–denaturing gradient gel electrophoresis(PCR–DGGE) and further16 S ribosomal DNA(r DNA) clone library analysis. The abundance of targeted genes for polycyclic aromatic hydrocarbon(PAH) degradation, nah Ac/phn Ac and C12O/C23 O, was tracked to define the metabolic ability of the in situ microbial community by Most Probable Number(MPN) PCR. The biosystem performed almost the same for treatment of both produced waters in terms of removals of chemical oxygen demand(COD) and PAHs. Sludge communities were closely associated with the respective influent bacterial communities(similarity 60%), while one sludge clone library was dominated by the Betaproteobacteria(38%) and Bacteriodetes(30%)and the other was dominated by Gammaproteobacteria(52%). This suggested that different influent and water quality have an effect on sludge community compositions. In addition, the existence of catabolic genes in sludge was consistent with the potential for degradation of PAHs in the treatment of both produced waters.     

海洋油气田沉积物产甲烷活性及微生物生态

海洋产甲烷古菌是与海洋中甲烷的产生、释放及天然气水合物的形成等密切相关的微生物类群.以产甲烷菌可能利用的底物为碳源,评价了海洋油气田沉积物的产甲烷活性.结果表明,海洋油气田沉积物对于H2/CO2、甲醇、一甲胺、三甲胺,在25℃和37℃两种温度条件下都具有较高的产甲烷活性,少数样品只在37℃对乙酸有产甲烷活性,说明在海洋油气田环境下,主要有氢营养型及甲基营养型发酵途径产生天然气甲烷.利用厌氧培养法对产甲烷菌进行了多样性解析,16S rRNA基因测序显示,优势产甲烷菌主要属于Methanosarcinales目的 Methanosarcinaceae科及Methanomicrobiales目的 Methanomicrobiaceae科,其中Methanococcoides、Methanogenium和Methanosarcina为油气田沉积物的主要菌属.