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.     

Absolute dominance of hydrogenotrophic methanogens in full-scale anaerobic sewage sludge digesters

Anaerobic digestion （AD） is gaining increasing attention due to the ability to covert organic pollutants into energy-rich biogas and, accordingly, growing interest is paid to the microbial ecology of AD systems. Despite extensive efforts, AD microbial ecology is still limitedly understood, especially due to the lack of quantitative information on the structures and dynamics of AD microbial communities. Such knowledge gap is particularly pronounced in sewage sludge AD processes although treating sewage sludge is among the major practical applications of AD. Therefore, we examined the microbial communities in three full-scale sewage sludge digesters using qualitative and quantitative molecular techniques in combination： denaturing gradient gel electrophoresis （DGGE） and real-time polymerase chain reaction （PCR）. Eight out of eleven bacterial sequences retrieved from the DGGE analysis were not affiliated to any known species while all eleven archaeal sequences were assigned to known methanogen species. Quantitative real-time PCR analysis revealed that, based on the 16S rRNA gene abundance, the hydrogenotrophic order Methanomicrobiales is the most dominant methanogen group （〉 94% of the total methanogen population） in all digesters. This corresponds well to the prevailing occurrence of the DGGE bands related to Methanolinea and Methanospirillum, both belonging to the order Methanomicrobiales, in all sludge samples. It is therefore suggested that hydrogenotrophic methanogens, especially Methanomicrobiales strains, are likely the major players responsible for biogas production in the digesters studied. Our observation is contrary to the conventional understanding that aceticlastic methanogens generally dominate methanogen communities in stable AD environments, suggesting the need for further studies on the dominance relationship in various AD systems.     

pH、ORP制约的产酸相发酵类型及顶级群落

由于利用效果存在一定差异，不同产酸相发酵类型的末端产物作为产甲烷细菌的底物，影响整个二相厌氧生物处理系统的稳定、高效运行。pH、ORP作为产酸相系统中的非生物因子，在一定程度上影响产酸相发酵类型的形成。本实验采用连续流搅拌槽式厌氧反应器（CSTR）考察并分析了特定pH、ORP组成对产酸相发酵类型决定情况，同时以种群生态学理论为基础，确立了受pH、ORP制约的不同发酵类型的顶级群落及优势种群。     

Microbial community structures in an integrated two-phase anaerobic bioreactor fed by fruit vegetable wastes and wheat straw

The microbial community structures in an integrated two-phase anaerobic reactor（ITPAR）were investigated by 16 S r DNA clone library technology. The 75 L reactor was designed with a 25 L rotating acidogenic unit at the top and a 50 L conventional upflow methanogenic unit at the bottom, with a recirculation connected to the two units. The reactor had been operated for 21 stages to co-digest fruit/vegetable wastes and wheat straw, which showed a very good biogas production and decomposition of cellulosic materials. The results showed that many kinds of cellulose and glycan decomposition bacteria related with Bacteroidales,Clostridiales and Syntrophobacterales were dominated in the reactor, with more bacteria community diversities in the acidogenic unit. The methanogens were mostly related with Methanosaeta, Methanosarcina, Methanoculleus, Methanospirillum and Methanobacterium; the predominating genus Methanosaeta, accounting for 40.5%, 54.2%, 73.6% and 78.7% in four samples from top to bottom, indicated a major methanogenesis pathway by acetoclastic methanogenesis in the methanogenic unit. The beta diversity indexes illustrated a more similar distribution of bacterial communities than that of methanogens between acidogenic unit and methanogenic unit. The differentiation of methanogenic community composition in two phases, as well as pH values and volatile fatty acid（VFA） concentrations confirmed the phase separation of the ITPAR. Overall, the results of this study demonstrated that the special designing of ITPAR maintained a sufficient number of methanogens, more diverse communities and stronger syntrophic associations among microorganisms, which made two phase anaerobic digestion of cellulosic materials more efficient.     

Effect of manganese oxide-modified biochar addition on methane production and heavy metal speciation during the anaerobic digestion of sewage sludge

Low organic matter content and high heavy metal levels severely inhibit the anaerobic digestion (AD) of sewage sludge. In this study, the effect of added manganese oxide-modified biochar composite (MBC) on methane production and heavy metal fractionation during sewage sludge AD was examined. The MBC could increase the buffering capacity, enhance the methane production and degradation of intermediate acids, buffer the pH of the culture, and stabilize the sewage sludge AD process. The application of MBC positively impacted methane production and the cumulative methane yield increased up to 121.97%, as compared with the control. The MBC addition can improve metal stabilization in the digestate. An optimum MBC dose of 2.36?g was recommended, which would produce up to 121.1?L/kg volatile solids of methane. After the AD process, even though most of the metals accumulated in the residual solids, they could be transformation from the bio-available fractions to a more stable fraction. The total organic- and sulfide-bound and residual fraction content at a 3?g dose of MBC that is 0.12?g/g dry matter were 51.06% and 35.11% higher than the control, respectively. The results indicated that the application of MBC could improve the performance of AD and promote stabilization of heavy metals in sewage sludge post the AD process.     

A comparative study on the alternating mesophilic and thermophilic two-stage anaerobic digestion of food waste

An alternating mesophilic and thermophilic two stage anaerobic digestion(AD) process was conducted. The temperature of the acidogenic(A) and methanogenic(M) reactors was controlled as follows: System 1(S1) mesophilic A-mesophilic M;(S2) mesophilic A-thermophilic M; and(S3)thermophilic A-mesophilic M. Initially, the AD reactor was acclimatized and inoculated with digester sludge. Food waste was added with the soluble chemical oxygen demand(SCOD) concentrations of41.4–47.0 g/L and volatile fatty acids of 2.0–3.2 g/L. Based on the results, the highest total chemical oxygen demand removal(86.6%) was recorded in S2 while S3 exhibited the highest SCOD removal(96.6%). Comparing S1 with S2, total solids removal increased by 0.5%; S3 on the other hand decreased by 0.1 % as compared to S1. However, volatile solids(VS) removal in S1, S2, and S3was 78.5%, 81.7%, and 79.2%, respectively. S2 also exhibited the highest CH4 content, yield, and production rate of 70.7%, 0.44 L CH4/g VSadded, and 1.23 L CH4/(L·day), respectively. Bacterial community structure revealed that the richness, diversity, evenness, and dominance of S2 were high except for the archaeal community. The terminal restriction fragments dendrogram also revealed that the microbial community of the acidogenic and methanogenic reactors in S2 was distinct. Therefore,S2 was the best among the systems for the operation of two-stage AD of food waste in terms of CH4production, nutrient removal, and microbial community structure.     

污水厂超声污泥厌氧消化性能的对比研究

利用超声技术分别对武汉市龙王嘴和汤逊湖两污水处理厂的浓缩污泥进行破解处理.采用半连续流中温厌氧消化实验,研究超声预处理对不同污水处理厂剩余污泥厌氧消化性能的影响.结果表明,与传统污泥消化工艺相比,超声预处理能明显提高污泥厌氧消化的日产气量和消化效率,使污泥产气更加稳定.并且相对于传统消化污泥,超声消化污泥的沉降性更好,...     

Impact of ozone assisted ultrasonication pre-treatment on anaerobic digestibility of sewage sludge

Impact of ultrasonication(ULS) and ultrasonication–ozonation(ULS-Ozone) pre-treatment on the anaerobic digestibility of sewage sludge was investigated with semi-continuous anaerobic reactors at solid retention time(SRT) of 10 and 20 days. The control, ULS and ULS-Ozone reactors produced 256, 309 and 348 m L biogas/g CODfedand the volatile solid(VS) removals were 35.6%, 38.3% and 42.1%, respectively at SRT of 10 days. At SRT of20 days, the biogas yields reached 313, 337 and 393 m L biogas/g CODfedand the VS removal rates were 37.3%, 40.9% and 45.3% in the control, ULS and ULS-Ozone reactors, respectively.ULS-Ozone pre-treatment increased the residual organic amount in the digested sludge.These soluble residual organics were found to contain macromolecules with molecular weights(MW) larger than 500 k Da and smaller polymeric products with MW around 19.4and 7.7 k Da. These compounds were further characterized to be humic acid-like substances with fluorescent spectroscopy analysis.     

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.     

不同基质生物炭对厌氧处理餐厨垃圾效能及微生态的影响

探究了剩余污泥（SS）、餐厨垃圾（FW）、玉米芯（CC）、甘蔗渣（BG）4种不同基质生物炭对厌氧生物处理餐厨垃圾效能的影响,对厌氧污泥的关键酶活性、微生物群落分布以及代谢途径等微生态进行了分析.结果表明,厌氧反应器分别加入4种生物炭后,COD平均去除率分别提高了29.49%、23.16%、29.42%、40.32%;傅里叶红外分析表明,投加SS生物炭组出水中羟基、酰胺基以及C-O-C伸缩振动峰减弱.4个厌氧反应器中厌氧污泥的乙酸激酶活性分别为0.40,0.42,0.96,0.98 μmol/g,表明投加CC与BG生物炭促进了餐厨垃圾的厌氧水解酸化过程;厌氧污泥胞外聚合物的蛋白质/多糖之比分别为0.415、0.56、1.89、2.8,投加CC、BG生物炭提高了污泥的稳定性.4个厌氧反应器中拟杆菌门、变形菌门、厚壁菌门为主要菌群,投加BG生物炭促进了变形菌门与厚壁菌门的生长;对于古细菌而言,甲烷杆菌属与甲烷丝菌属为优势种群,SS组的甲烷杆菌属丰度最高（53.48%）,而BG组中甲烷丝菌属丰度最高（42.72%）.KEGG功能分析表明古菌及细菌均以碳水化合物代谢、氨基酸代谢为主;而投加BG与SS生物炭后,微生物膜运输水平得到了提高.