The key role of inoculated sludge in fast start-up of sequencing batch reactor for the domestication of aerobic granular sludge

Two types of inoculated sludges, granular sludge that had been stored at-20°C and activated sludge, were investigated for the domestication of aerobic granular sludges(AGSs in sequencing batch reactors(SBRs). The results showed that using the stored granular sludge as inoculation sludge could effectively shorten the domestication time of AGS and yielded mature granular sludge after 22 days of operation. The AGS domesticated by stored granular sludge had better biomass and sedimentation properties; its MLSS and SVI reached8.55 g/L and 35.27 mL/g, respectively. The removal efficiencies for chemical oxygen demand(COD), ammonium nitrogen(NH_4~+-N) and total phosphorus(TP) reached 90.76%, 97.39% and 96.40%, respectively. By contrast, 54 days were needed to obtain mature granules using activated sludge. The microbial community structure was probed by using scanning electron microscopy(SEM) and high-throughput sequencing. The results showed that the diversity of the microbial community in mature granules was reduced when stored granular sludge rather than activated sludge was employed as inoculation sludge, and the dominant microbes were changed. The dominant species in mature granules domesticated using stored granular sludge were Zoogloea, Acidovorax and Tolumonas at the genus classification level, while the dominant species were Zoogloea and TM7-genera in granules developed from activated sludge.     

Effect of inorganic carbon on anaerobic ammonium oxidation enriched in sequencing batch reactor

The present lab-scale research reveals the enrichment of anaerobic ammonium oxidation microorganism from methanogenic anaerobic granular sludge and the effect of inorganic carbon(sodium bicarbonate)on anaerobic ammonium oxidation.The enrichment of anammox bacteria was carried out in a 7.0-L sequencing batch reactor(SBR)and the effect of bicarbonate on anammox was conducted in a 3.0-L SBR.Research results,especially the biomass,showed first signs of anammox activity after 54 d cultivation with synthetic w...     

Effect of seed sludge on characteristics and microbial community of aerobic granular sludge

Aerobic granular sludge was cultivated by using different kinds of seed sludge in sequencing batch airlift reactor.The influence of seed sludge on physical and chemical properties of granular sludge was studied;the microbial community structure was probed by using scanning electron microscope and polymerase chain reaction-denaturing gradient gel electrophoresis(PCR-DGGE).The results showed that seed sludge played an important role on the formation of aerobic granules.Seed sludge taken from beer wastewater treatment plant(inoculum A) was more suitable for cultivating aerobic granules than that of sludge from municipal wastewater treatment plant(inoculum B).Cultivated with inoculum A, large amount of mature granules formed after 35 days operation, its SVI reached 32.75 mL/g, and SOUR of granular sludge was beyond 1.10 mg/(g·min).By contrast, it needed 56 days obtaining mature granules using inoculum B.DGGE profiles indicated that the dominant microbial species in mature granules were 18 and 11 OTU when inoculum A and B were respectively employed as seed sludge.The sequencing results suggested that dominant species in mature granules cultivated by inoculum A were Paracoccus sp., Devosia hwasunensi, Pseudoxanthomonas sp., while the dominant species were Lactococcus raffinolactis and Pseudomonas sp.in granules developed from inoculum B.     

Community analysis of ammonia and nitrite oxidizers in start-up of aerobic granular sludge reactor

A lab-scale sequencing batch reactor (SBR) was setup and the aerobic granular sludge was successfully incubated by using anaerobic granular sludge as seed sludge. Nitrogen was partially removed by simultaneous nitrification and denitrification (SND) via nitrite with free ammonia (FA) of about 10 mg/L. The denaturing gradient gel electrophoresis (DGGE) method was used to investigate community structure of α-Proteobacteria, β-Proteobacteria, ammonia oxidizing bacteria (AOB), and Nitrospira populations during start-up. The population sizes of bacteria, AOB and Nitrospira were examined using real-time PCR method. The analysis of community structure and Shannon index showed that stable structure of AOB population was obtained at the day 35, while the communities of α-Proteobacteria, β-Proteobacteria, and Nitrospira became stable after day 45. At stable stage, the average cell densities were 1.1×1012, 2.2×1010 and 1.0×1010 cells/L for bacteria, AOB and Nitrospira, respectively. The relationship between characteristics of nitrifying bacteria community and nitrogenous substrate utilization constant was discussed by calculating Pearson correlation. Certain correlation seemed to exist between population size, biodiversity, and degradation constant. And the influence of population size might be greater than that of biodiversity.     

Robustness of anammox granular sludge treating low-strength sewage under various shock loadings: Microbial mechanism and little N2O emission

With the increasing application of anammox for the treatment of high-strength industrial wastewater, application of anammox in municipal sewage has been gaining more attention. Sludge granulation in particular enhances the enrichment and retention of anammox bacteria in municipal sewage treatment systems. However, the performance of granular sludge under continuous and varying hydraulic loading shock remains little understood. In this study, the robustness of anammox granular sludge in treating low-strength municipal sewage under various shock loadings was investigated. Results showed that an upflow anaerobic sludge blanket (UASB) reactor with anammox granules performed well, with anammox specific activity up to 0.28?kg?N/kg VSS/day and anti-loading shock capability up to 187.2?L/day during the 8-month testing period. The accumulation rate of N₂O (< 0.01?kg?N/kg VSS/day) in the liquid phase was seven times higher than that of the gas phase, which could be mainly attributed to the incomplete denitrification and insufficient carbon source. However, only a small part of the produced N₂O escaped into the atmosphere. High-throughput sequencing and molecular ecological network analyses also identified the bacterial diversity and community structure, indicating the potential resistance against loading shock. The composition and structural analyses showed that polysaccharides were an important functional component in the tightly bound extracellular polymeric substances (TB-EPS), which was the major EPS layer of anammox granules. Scanning electron microscopy (SEM) also showed that the gaps in between the anammox-clusters in the granules inhibit the flotation of the sludge and ensure efficient settling and retention of anammox granules.     

Bioaugmentation with a pyridine-degrading bacterium in a membrane bioreactor treating pharmaceutical wastewater

The bacterial strain Paracoccus denitrificans W12, which could utilize pyridine as its sole source of carbon and nitrogen, was added into a membrane bioreactor （MBR） to enhance the treatment of a pharmaceutical wastewater. The treatment efliciencies investigated showed that the removal of chemical oxygen demand, total nitrogen, and total phosphorus were similar between bioaugmented and non-bioaugmented MBRs, however, significant removal of pyridine was obtained in the bioaugmented reactor. When the hydraulic retention time was 60 hr and the influent concentration of pyridine was 250-500 mg/L, the mean effluent concentration of pyridine without adding W12 was 57.2 mg/L, while the pyridine was degraded to an average of 10.2 mg/L with addition of W12. The bacterial community structure of activated sludge during the bioaugmented treatment was analyzed using polymerase chain reaction-denaturing gradient gel electrophoresis （PCR-DGGE）. The results showed that the W12 inoculum reversed the decline of microbial community diversity, however, the similarity between bacterial community structure of the original sludge and that of the sludge after bioaugmentation decreased steadily during the wastewater treatment. Sequencing of the DNA recovered from DGGE gel indicated that sp., Sphingobium sp., Comamonas sp., and Hyphomicrobium sp. were the dominant organisms in time sequence in the bacterial community in the bioaugmented MBR. This implied that the bioaugmentation was affected by the adjustment of whole bacterial community structure in the inhospitable environment, rather than being due solely to the degradation performance of the bacterium added.     

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.     

Revealing the anaerobic acclimation of microbial community in a membrane bioreactor for coking wastewater treatment by Illumina Miseq sequencing

The dynamic change of microbial community during sludge acclimation from aerobic to anaerobic in a MBR for coking wastewater treatment was revealed by Illumina Miseq sequencing in this study. The diversity of both Bacteria and Archaea showed an increase–decrease trajectory during acclimation, and exhibited the highest at the domestication interim. Ignavibacteria changed from a tiny minority(less than 1%) to the dominant bacterial group(54.0%) along with acclimation. The relative abundance of Betaproteobacteria kept relatively steady, as in this class some species increased coupled with some other species decreased during acclimation. The dominant Archaea shifted from Halobacteria in initial aerobic sludge to Methanobacteria in the acclimated anaerobic sludge. The dominant bacterial and archaeal groups in different acclimation stages were indigenous microorganisms in the initial sludge, though some of them were very rare. This study supported that the species in"rare biosphere" might eventually become dominant in response to environmental change.     

Influence of oxytetracycline on the structure and activity of microbial community in wheat rhizosphere soil

The microbial community composition in wheat rhizosphere was analyzed by detecting colony forming units (CFUs) in agar plates. The total CFUs in rhizosphere were 1.04 109/g soil with 9.0 108/g bacteria, 1.37 108/g actinomyces and 3.6 106/g fungi. The 10 dominant bacteria were isolated from wheat rhizosphere and were grouped into genus Bacillus according to their full length 16S rRNA gene sequences. Although belonging to the same genus, the isolated strains exhibited di erent sensitivities to oxytetracycline. When a series of the rhizosphere soil was exposed under various concentrations of oxytetracycline, the microbial community structure was highly a ected with significant decline of CFUs of bacteria and actinomyces (22.2% and 31.7% at 10 mg/kg antibiotic, respectively). This inhibition was clearly enhanced with the increase exposure dosage of antibiotic and could not be eliminated during 30 d incubation. There was no obvious influence of this treatment on fungi population. Among the four soil enzymes (alkaline phosphatase, acidic phosphatase, dehydrogenase and urease), only alkaline phosphatase was sensitive to oxytetracycline exposure with 41.3% decline of the enzyme activity at 10 mg/kg antibiotic and further decrease of 64.3%–80.8% when the dosage over 30 mg/kg.     

The difference of morphological characteristics and population structure in PAO and DPAOgranular sludges

We examined how long-term operation of anaerobic–oxic and anaerobic–anoxic sequencing batch reactors(SBRs) affects the enhanced biological phosphorus removal(EBPR)performance and sludge characteristics. The microbial characteristics of phosphorus accumulating organism(PAO) and denitrifying PAO(DPAO) sludge were also analyzed through a quantitative analysis of microbial community structure. Compared with the initial stage of operation characterized by unstable EBPR, both PAO and DPAO SBR produced a stable EBPR performance after about 100-day operation. From day 200 days(DPAO SBR)and 250 days(PAO SBR) onward, sludge granulation was observed, and the average granule size of DPAO SBR was approximately 5 times larger than that of PAO SBR. The DPAO granular sludge contained mainly rod-type microbes, whereas the PAO granular sludge contained coccus-type microbes. Fluorescence in situ hybridization analysis revealed that a high ratio of Accumulibacter clade I was found only in DPAO SBR, revealing the important role of this organism in the denitrifying EBPR system. A pyrosequencing analysis showed that Accumulibacter phosphatis was present in PAO sludge at a high proportion of 6%,whereas it rarely observed in DPAO sludge. Dechloromonas was observed in both PAO sludge(3.3%) and DPAO sludge(3.2%), confirming that this organism can use both O_2 and NO_3~- as electron acceptors. Further, Thauera spp. was identified to have a new possibility as denitrifier capable of phosphorous uptake under anoxic condition.     

SBR反应器厌氧氨氧化系统启动过程中细菌群落结构的变化

以白洋淀岸边带沉积物为接种污泥,启动了SBR厌氧氨氧化反应器.对反应器启动过程中的进出水水质进行了连续监测,并采用PCRDGGE、定量PCR和基因测序等分子生物学技术研究了系统内总细菌和厌氧氨氧化(ANAMMOX)细菌群落结构随培养时间的变化规律.结果表明:在启动过程中,总微生物菌群动态变化水平为26.6%~50.5%;微生物多样性先变小后增大,优势菌种得到重新分布;ANAMMOX细菌的群落结构变得单一化,最后系统的优势ANAMMOX细菌是Brocadia属.富集培养阶段SBR系统中ANAMMOX细菌的最大生物量达到了1.73×109copies·g~(-1)干污泥,而且总氮的去除率最高达到约82%.     

厌氧氨氧化细菌富集培养过程微生物结构与功能解析

为了研究厌氧氨氧化细菌富集培养过程中微生物结构与功能的关系,本研究采集升流式污泥床反应器（Upflow anaerobic sludge bed,UASB）中厌氧氨氧化反应启动与驯化过程的样品,结合传统微生物培养方法和分子生物学手段,对启动过程中的氮素转化功能基因、异养菌演替及菌群演替进行了定性与定量分析.结果表明,随着总氮去除率从1.39%升高至79.62%,厌氧氨氧化细菌功能基因hzo大量富集,且统计分析结果表明nirS是影响脱氮效果的关键因素.微生物菌群在门水平上主要优势菌门为厌氧氨氧化细菌所在的浮霉菌门（Planctomycetes）,在属水平上的优势菌属为Candidatus Kuenenia,丰度从4.75%升高至48.77%.这些都表明厌氧氨氧化细菌的成功富集是实现脱氮效果的主要因素.尽管进水无外加碳源,但异养菌始终占有一定的比例,与厌氧氨氧化细菌存在共生现象,却在功能上发生演替.网络分析和基于KEGG数据库的功能预测结果表明,厌氧氨氧化细菌Candidatus Kuenenia和异养菌Ignavibacterium、Gp4呈显著正相关（p<0.05）,这些异养菌通过降解或合成胞外物质,分泌次级代谢物质与厌氧氨氧化细菌相互作用,进而影响厌氧氨氧化细菌的活性与生长.     

厌氧氨氧化脱氮除碳功能菌群结构及代谢途径

为明确不同有机物浓度(50～150mg/L)和竹炭同时存在下厌氧氨氧化颗粒污泥系统的脱氮除碳功能菌群结构及代谢途径差异,采用宏基因组测序技术对其微生物分布规律和碳氮代谢基因表达进行了研究.结果表明,当COD浓度为50,150mg/L,添加竹炭显著提升了厌氧氨氧化菌(AnAOB)的相对丰度,Candidatus_Kuen...     

基于高通量测序的ABR厌氧氨氧化反应器各隔室细菌群落特征分析

为明确厌氧折流板反应器(ABR)稳定运行厌氧氨氧化反应后各隔室微生物群落结构特征,本文采用Miseq高通量测序分析技术,对ABR厌氧氨氧化反应器5个隔室的微生物分布规律进行了研究,结果表明,ABR反应器中脱氮微生物多样性较为丰富,变形菌门(Proteobacteria)占11.66%~20.28%,浮霉菌门(Planctomycetes)占2.18%~7.94%,硝化螺旋菌门(Nitrospirae)占0.19%~6.30%.其中,在ABR反应器中变形菌门占据主导地位,主要包含Rhodoplanes、Dok59、Rubrivivax和Bdellovibrio等菌属,浮酶菌门次之,主要包含Candidatus brocadia和Candidatus kuenenia菌属.从第1~5隔室,污泥表观红色逐渐减退,趋向于灰黑色,Chao、ACE、Shannon、Simpson指数均表明微生物群落丰富度逐渐增加,且变形菌门微生物逐渐增加,而浮霉菌门微生物逐渐降低,这与基质的降解和功能微生物的富集规律相一致.     

控制游离氨实现单级自养生物脱氮的研究

通过实时调控SBR反应器内的游离氮浓度的控制策略,实现以亚硝化作用和厌氧氨氧化作用协同的单级自养生物脱氮工艺.实验分成亚硝酸菌富集和厌氧氨氧化菌混合接种2个阶段,SBR内的温度始终保持在(31±2)℃.亚硝酸菌富集阶段,pH值稳定在7.8左右,通过调节进水氩氮浓度(56～446 mg·L-1)实现FA浓度的变化,从而实...     

Fast start-up anammox process using Acyl-homoserine lactones (AHLs) containing supernatant

N-dodecanoyl homoserine lactone (C₁₂-HSL) was detected in the supernatant of an anammox granular sludge reactor (AGSR). C₁₂-HSL could enhance the specific anammox activity of anammox biomass. Adding C₁₂-HSL-containing AGSR supernatant into the continuously stirred tank reactors reduced the start-up time of the anammox process from 80 to 66 days. Moreover, the nitrogen loading rate was also enhanced to 1.6 times that of the control reactor. AHLs could increase the secretion of extracellular polymeric substances and anammox obtained better enrichment with the addition of AHLs-containing AGSR supernatant. Denaturing gradient gel electrophoresis analysis further revealed that AHLs played a role in mediating microbial community parameters. In conclusion, adding AHL-containing supernatant could be an effective and economical way to accelerate the start-up of anammox.     

长三角地区MBBR泥膜复合污水厂低温季节微生物多样性分析

为表征长三角地区采用MBBR泥膜复合工艺（hybrid-MBBR）的污水厂在低温季节的微生物群落变化,得出微生物分布规律,采用Illumina MiSeq高通量测序对该区域5座市政污水处理厂进行研究,对好氧区活性污泥及悬浮载体生物膜微生物群落结构进行了分析.结果表明,同污水厂悬浮载体生物膜微生物物种数低于同系统活性污泥,且物种分布更不均匀.悬浮载体的投加可提升系统微生物多样性,但同时进水及运行方式对系统微生物群落组成具有一定选择性.各污水厂相对丰度较高的菌属主要有Nitrospira、Mycobacterium、Defluviicoccus、Hyphomicrobium和Macellibacteroides等,悬浮载体的投加极大程度上强化了优势硝化菌属Nitrospira的富集.核算悬浮载体中硝化细菌生物量占系统中总量的86.12%~95.36%;各污水厂好氧区悬浮载体中均检测到一定相对丰度的反硝化菌群,结合沿程及小试结果确认好氧区悬浮载体生物膜上发生了显著的同步硝化反硝化（SND）现象,强化了系统TN去除.     

热水解时间对污泥厌氧消化系统微生物群落结构影响分析

为评估热水解时间对北京市大兴区某污水处理厂污泥厌氧消化系统微生物群落结构的影响,利用Illumina MiSeq高通量测序方法,分析了不同热水解时间（15、30和45 min）对初沉污泥和剩余活性污泥厌氧消化系统中微生物群落结构及其多样性的影响因素.结果表明,消化污泥优势类群主要分布在厚壁菌门（Firmicutes）、阴沟单胞菌门（Cloacimonadota）、绿弯菌门（Chloroflexi）和同力菌门（Synergistota）,相对丰度之和超过60%,相对丰度最高菌属为W5,占比为20.8%~54.5%,表现为少数优势物种的高丰度特征.热水解污泥厌氧消化过程中,高挥发性脂肪酸和氨氮浓度导致嗜乙酸产甲烷菌相对丰度减少,嗜氢产甲烷途径多于嗜乙酸产甲烷途径.环境因子关联分析结果显示,消化进泥可溶蛋白质、消化进泥pH值、消化出泥氨氮和热水解时间是影响微生物群落结构的4个主要环境因子,其中消化出泥氨氮对产甲烷菌属的影响最大,呈负相关关系.热水解时间与Chao指数和Shannon指数均呈负相关,较长热水解时间不利于提高厌氧消化过程微生物菌群丰富度和多样性.