两次污泥颗粒化过程中微生物群落的动态变化

利用序批式反应器(sequencing batch reactor,SBR)培养好氧颗粒污泥(aerobic granular sludge,AGS),在此期间发生了AGS破碎现象,后经培养,破碎污泥再次变为成熟的AGS.因此,采用Illumina Mi Seq PE300高通量测序技术研究了两次污泥颗粒化过程中微生物群落结构变化的差异,以期揭示有利于AGS形成的优势菌属;此外,利用实时定量聚合酶链式反应(quantitative polymerase chain reaction,q PCR)探究了两次污泥颗粒化过程中硝化微生物的动态变化.结果表明,在两次污泥颗粒化过程中,胞外蛋白质和胞外多糖的含量均增加;氨氧化古菌(ammonia oxidizing archaea,AOA)在第一次污泥颗粒化过程以及AGS成熟过程丰度增加,氨氧化细菌(ammonia oxidizing bacteria,AOB)虽然在第一次污泥颗粒化过程中丰度降低,但是在AGS培养过程中其丰度都一直高于AOA;微生物群落多样性随着AGS的形成而降低;变形菌门(Proteobacteria)相对丰度明显增加,分别增加了12.29%和5.90%;某些属于变形菌门的属其相对丰度也增加,其中,Candidatus Competibacter在两次污泥颗粒化过程中相对丰度增加最明显,并且在成熟的AGS中呈现很高的相对丰度,达到14.20%.总的来说,胞外蛋白质和胞外多糖含量的增加,可能促进了污泥颗粒化;AOA和AOB可能共同参与了AGS的氨氧化作用;Ca.Competibacter的富集可能有利于AGS的形成.     

Single-cell sequencing and mini-sequencing for preimplantation genetic diagnosis

There is increasing interest in the use of preimplantation genetic diagnosis (PGD) as an alternative to routine prenatal diagnosis. However, the costs associated with development and testing of new PGD protocols have forced some PGD centres to limit the number of diseases for which PGD is offered. One of the main factors in the design of new protocols, which affects cost and accuracy, is the choice of the mutation-detection technique. We have assessed the reliability of DNA sequencing and mini-sequencing for clinical diagnosis at the single-cell level and have found them to be rapid and accurate. Extensive optimisation for individual mutations is not usually necessary when employing these versatile techniques and consequently a smaller investment of time and resources should be required during development of new protocols. Additionally, we report single-cell protocols for the diagnoses of cystic fibrosis, sickle cell anaemia and β-thalassaemia, which utilise mini-sequencing. Unlike most mutation-detection techniques, mini-sequencing permits analysis of very small DNA fragments. Small amplicons experience low allele dropout (ADO) rates, and consequently this approach could potentially improve the reliability of PGD. Copyright © 2003 John Wiley & Sons, Ltd.     

低浓度铬对SBR中微生物抑制影响研究

研究了低浓度铬(三价和六价)对SBR生物系统的抑制影响,考察了两种不同SBR工艺(传统工艺和分段进水工艺)在处理含低浓度铬废水过程中常规的出水水质和活性污泥性状的变化,以及微生物群落的变迁.研究结果表明,在进水总铬(Cr(III)∶Cr(VI)=4∶1)浓度为0.5mg·L-1的条件下,传统工艺和分段进水工艺的氨氮去除率由99%分别下降至70%和65%,同时,磷酸盐去除率也由99%分别下降至51%和43%.当进水中总铬浓度达到1 mg·L-1时,分段进水工艺SBR系统的氨氮和磷酸盐去除率最终分别下降至44%和37%.此外,多糖和蛋白质的分泌量变化分别呈下降和上升趋势.高通量测序结果表明,活性污泥细菌群落丰富度和多样性受到了铬离子的影响,并且Nitrospira、Acidobacteria、Planctomycetes、Cyanobacteria和Candidatus_Accumulibacter等脱氮除磷功能菌种的生长都受到了一定程度的抑制,也与被抑制的SBR系统脱氮除磷去除率下降的宏观现象相吻合.     

能源植物修复镉污染土壤对根际细菌网络结构的影响

选取油脂类能源植物大豆和碳水化合物类能源植物玉米,采用高通量测序方法研究大豆、玉米修复Cd污染土壤过程中根际土壤细菌群落组成,基于高通量测序数据采用分子生态网络分析细菌相互作用.结果表明,50 mg·kg-1Cd污染土壤中两种植物根部Cd浓度和积累量最高,转移系数TF分别为玉米0.78和大豆0.35.基于细菌16S r RNA基因的群落分析表明,大豆、玉米根际土壤细菌主要包括Proteobacteria(变形菌门)、Acidobacteria(酸杆菌门)31个门细菌组成,大豆、玉米种植均能影响土壤细菌群落组成,能影响Candidate division TM7 norank、Acidimicrobiales norank、Sphingomonas等丰度.分子生态网络分析表明种植大豆和玉米增加了细菌之间的相互作用,导致其网络结构更为复杂,关键细菌从不种植物处理的1个增加到种植大豆的6个和种植玉米的10个.     

Distribution of bacterial communities across plateau freshwater lake and upslope soils

Microorganisms are involved in a variety of biogeochemical processes in natural environments. The differences between bacterial communities in freshwaters and upslope soils remain unclear. The present study investigated the bacterial distribution in a plateau freshwater lake, Erhai Lake (southwestern China), and its upslope soils. Illumina MiSeq sequencing illustrated high bacterial diversity in lake sediments and soils. Sediment and soil bacterial communities were mainly composed of Proteobacteria, Acidobacteria, Actinobacteria, Bacteroidetes, Chloroflexi and Planctomycetes. However, a distinctive difference in bacterial community structure was found between soil and sediment ecosystems. Water content, nitrogen and pH affected the distribution of the bacterial community across Erhai Lake and its upslope soils. Moreover, the soil bacterial community might also be shaped by plant types. This work could provide some new insights into plateau aquatic and terrestrial microbial ecology.     

Enhancing nitrogen removal from low carbon to nitrogen ratio wastewater by using a novel sequencing batch biofilm reactor

Removing nitrogen from wastewater with low chemical oxygen demand/total nitrogen (COD/TN) ratio is a difficult task due to the insufficient carbon source available for denitrification. Therefore, in the present work, a novel sequencing batch biofilm reactor (NSBBR) was developed to enhance the nitrogen removal from wastewater with low COD/TN ratio. The NSBBR was divided into two units separated by a vertical clapboard. Alternate feeding and aeration was performed in the two units, which created an anoxic unit with rich substrate content and an aeration unit deficient in substrate simultaneously. Therefore, the utilization of the influent carbon source for denitrification was increased, leading to higher TN removal compared to conventional SBBR (CSBBR) operation. The results show that the CSBBR removed up to 76.8%, 44.5% and 10.4% of TN, respectively, at three tested COD/TN ratios (9.0, 4.8 and 2.5). In contrast, the TN removal of the NSBBR could reach 81.9%, 60.5% and 26.6%, respectively, at the corresponding COD/TN ratios. Therefore, better TN removal performance could be achieved in the NSBBR, especially at low COD/TN ratios (4.8 and 2.5). Furthermore, it is easy to upgrade a CSBBR into an NSBBR in practice.     

Effect of wastewater COD/N ratio on aerobic nitrifying sludge granulation and microbial population shift

The effect of COD/N ratio on the granulation process and microbial population succession was investigated. Four identical sequencing batch reactors, R1, R2, R3 and R4, were operated with various initial COD/N ratios ranging from 0/200 to 800/200 (m/m). Ethanol was fed as the source of COD. Aerobic granules were successfully cultivated in R2 and R3, operating with the COD/N ratio of 200/200 and 400/200, respectively. Scanning electron microscope observations indicated that short rod-shaped and spherical bacteria were dominant in R2, while granules produced in R3 were surrounded with a large amount of filamentous bacteria. The average specific nitritation rate in R2 and R3 were 0.019 and 0.008 mg N/(mg MLVSS.hr), respectively. Fluorescence in situ hybridization results demonstrated that nitrifying bacteria population was enriched remarkably in R2. It indicated that nitrification ability and nitrifying bacteria population were enriched remarkably at low COD/N ratio. However, no granules were formed in R1 and R4 which might attribute to either limited or excessive extracellular polymeric substances production. This study contributed to a better understanding of the role of COD/N ratio in nitrifying sludge granulation.     

Biodegradation of p-cresol by aerobic granules in sequencing batch reactor

The cultivation of aerobic granules in sequencing batch reactor for the biodegradation of p-cresol was studied. The reactor was started with 100 mg/L of p-cresol. Aerobic granules first appeared within one month of start up. The granules were large and strong and had a compact structure. The diameter of stable granules was in the range of 1-5 mm. The integrity coefficient and granules density was found to be 96% and 1046 kg/m³, respectively. The settling velocity of granules was found to be in the range of 2× 10^-2-6× 10^-2 m/sec. The aerobic granules were able to degrade p-cresol upto 800 mg/L at a removal efficiency of 88%. Specific p-cresol degradation rate in aerobic granules followed Haldane model for substrate inhibition. High specific p-cresol degradation rate up to 0.96 g p-cresol/(g VSS. day) were sustained upto p-cresol concentration of 400 mg/L. Higher removal efficiency, good settling characteristics of aerobic granules, makes sequencing batch reactor suitable for enhancing the microorganism potential for biodegradation of inhibitory compounds.     

活性污泥低温氨氧化功能的驯化与潜力研究

以强化低温污水生物处理系统的氨氮(NH4+-N)氧化功能为目标,采用序批式活性污泥反应器(SBR),通过逐步提高进水NH4+-N浓度的方式,在(15±1)℃的条件下对城镇污水处理厂的好氧活性污泥进行了驯化培育,并就其对生活污水和高氨氮废水的NH4+-N去除潜力进行了测试.结果表明,在(15±1)℃下,好氧活性污泥经适当驯化可获得良好的氨氧化能力.在初始NH4+-N浓度为46mg·L-1左右时,其NH4+-N去除速率和亚硝态氮(NO2--N)生成速率分别可达54.26g·kg-·1d-1(以MLSS计,下同)和29.07g·kg-·1d-1(以MLSS计,下同)左右.对NH4+-N浓度为47.19mg·L-1左右的城镇污水,其NH4+-N去除率可高达85%以上.初始NH4+-N浓度分别为91.01mg·L-1和163.37mg·L-1左右时,其最高NH4+-N去除速率分别可达52.54g·kg-·1d-1和111.97g·kg-·1d-1,具有处理高氨氮废水的潜力.     

曝气量及COD浓度对SBBR同步硝化反硝化效能的影响

采用悬浮球形填料,以人工模拟生活污水为原水,研究了曝气量及进水COD浓度对序批式膜生物反应器(SBBR)内同步硝化反硝化(SND)效能的影响。试验结果表明,在0.025~0.070 m3/h的曝气量范围内,SBBR的COD及NH4+-N去除率都能达到90%左右,COD和NH4+-N的降解过程具有时序性,即曝气初期以COD降解为主,待COD降解基本完成后,NH4+-N降解速率才明显提高。SBBR内同步硝化反硝化效率与曝气量成反比,即曝气量越低,TN去除率越高;当曝气量为0.025 m3/h时,TN去除效率最高,达到73.26%。在200~700 mg/L的进水COD浓度范围内,COD去除率始终维持在90%左右,TN去除率随着进水COD浓度增加呈现先升高后降低的变化趋势。