甲烷厌氧氧化协同硝酸盐还原菌群驯化及其群落特征

伴随硝酸盐还原的甲烷厌氧氧化是协同减少环境中硝酸盐及甲烷的有效途径.利用实验室废水处理厌氧污泥、污水处理厂厌氧污泥和填埋场覆土驯化富集硝酸盐还原型甲烷厌氧氧化菌群.考察菌群的甲烷氧化效果,结果发现接种污水处理厂厌氧污泥体系甲烷转化量最大,为0.05 mg·d-1.微生物群落结构分析显示,该体系中甲烷微菌和甲烷八叠球菌是甲烷氧化菌,假单胞菌、梭状芽胞杆菌和热单胞菌参与了硝酸盐的还原反应.硝酸盐的量影响甲烷的转化率及菌群结构.当硝酸盐浓度为200 mg·L-1时,体系中的硝酸盐还原菌为假单胞菌和梭状芽胞杆菌;浓度增加至500 mg·L-1时,硝酸盐还原菌则是假单胞菌和热单胞菌.同时,甲烷转化率增加34.7%.研究结果为该菌群应用于含甲烷废气与含硝酸盐废水的协同处理提供科学依据.     

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.     

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.     

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.     

硫酸盐还原型甲烷厌氧氧化菌群驯化及其群落特征

甲烷的温室效应是二氧化碳的26倍,高浓度硫酸盐废水对水体、土壤和植物均有危害.硫酸盐为氧化剂的甲烷厌氧氧化是减少甲烷的主要途径之一.本研究以硫酸盐作为电子受体,驯化培养硫酸盐还原型甲烷厌氧氧化菌群,采用PCR-DGGE技术分析细菌和古菌菌群多样性和群落结构特征,并对其中的优势菌进行系统发育分析.DGGE指纹图谱结果表明,硫酸盐的加入使微生物群落结构和优势种群数量发生了明显的改变,其增强了甲烷氧化古菌和硫酸盐还原细菌的丰度,加入硫酸盐驯化的菌群,其细菌群落多样性增加而古菌群落多样性略微减少.典型条带测序结果显示,驯化后菌群的优势菌种主要包括螺旋体门(Spirochaetes),除硫单胞菌目(Desulfuromonadales)、甲烷八叠球菌目(Methanosarcinales)和甲烷丝状菌属(Methanosaeta)等.驯化菌群的甲烷厌氧氧化研究结果显示,甲烷厌氧氧化的同时伴随硫酸盐的还原,甲烷的氧化产物为二氧化碳,硫酸盐的转化产物为硫化氢和硫单质.     

AFMBR处理生活污水的功能菌群特性研究

厌氧流化床膜生物反应器(AFMBR)作为一种低耗产能的高效厌氧反应器,在处理生活污水中有着巨大的潜力.本研究主要利用宏基因组测序技术对AFMBR系统内的微生物菌群进行探究,结果表明:与接种菌群相比,AFMBR经过一段时间的连续运行后,在属水平上古菌优势菌属由接种时的甲烷囊菌属变为甲烷八叠球菌属、甲烷杆菌属,细菌的整体菌属结构发生了较大变化;在种水平上,系统内不存在较明显的优势菌种.从相对丰度比例≥1%的菌种来看:水解发酵菌群产氢产乙酸菌群产甲烷菌群,但各菌群之间相对丰度的差距较小.从基因水平来看,系统内与碳水化合物代谢、氨基酸代谢、能量代谢相关的基因丰度较高,二氧化碳、乙酸转化为甲烷是系统产甲烷的主要途径.     

乳制品废水中细菌的分离培养

本文通过微生物分离技术和一些微生物特定的鉴定方法，对乳制品综合废水中细菌进行不同温度的增菌培养，分离，纯化对比试验及显微镜观察，根据各菌株的个体，群体形态和生理生化实验鉴定到菌属，实验结果可见，微球菌属，黄杆菌属，产碱杆菌属，假单胞菌属为乳制品废水的优势菌群，并且培养温度３０℃为最适温度。     

ABR处理综合印染废水中细菌群落结构研究

印染废水是公认的难降解工业废水之一,以细菌菌群为主的厌氧生物污泥决定印染废水的脱色和解毒效果,但是基于厌氧折流板反应器(ABR)处理综合印染废水工程污泥细菌群落结构的研究鲜见报道。该文基于Miseq高通量测序分析技术,对长期稳定运行的ABR处理单元6个隔室的微生物菌群分布进行研究。结果表明,ABR处理单元中微生物菌群多样性较为丰富,且各隔室间存在较大差异。前5个隔室中优势细菌门类为变形菌门(Proteobacteria)、拟杆菌门(Bacteroidetes)、螺旋体门(Spirochaetae)和厚壁菌门(Firmicutes),而第6隔室为变形菌门(Proteobacteria)、绿弯菌门(Chloroflexi)和互养菌门(Synergistetes);主要属为螺旋体属(Spirochaetes_norank)、陶厄氏菌属(Thauera)、未命名菌属(vadinBC27_wastewater-sludge_group和Sh765B-TzT-29_norank)、长绳菌属(Longilinea)、副球菌属(Paracoccus)、利恩氏热杆菌属(Thermovirga)、丛毛单胞菌属(Comamonas)和假单胞菌属(Pseudomonas),大多数属与印染废水的脱色和降解过程有关。该结果为综合印染废水污染物的去除机制和运营提供了理论基础。     

不同原料厌氧发酵及其微生物种群的研究

使用PCR-DGGE技术,对以鸡粪、猪粪、牛粪、秸秆为发酵原料的发酵体系的微生物群落多样性进行了研究.在发酵不同时间取样,进行了日产甲烷量、日产甲烷浓度的变化分析和DGGE分析.结果表明,日产甲烷量整体趋势为猪粪>鸡粪>秸秆>牛粪,日平均产甲烷量分别为2.67、2.24、0.99、0.49L;猪粪、鸡粪、秸秆的日产甲烷浓度在整个发酵周期大多可维持在50%以上,牛粪的日产甲烷浓度大部分时间低于30%;细菌的优势菌群有拟杆菌属(Bacteroidetes)、密螺旋体属(Treponema)、厌氧绳菌科(Anaerolineaceae)等,新增优势菌群有梭菌属(Clostridium)、脱硫叶菌属(Desulfobulbus)、毛螺旋菌科(Lachnospiraceae)、醋弧菌属(Acetivibrio);古菌的优势菌群有甲烷鬃菌属(Methanosaeta)、甲烷八叠球菌属(Methanosarcina)、甲烷八叠球菌目(Methanosarcinales),新增优势菌群有斯氏甲烷球菌属(Methanosphaera stadtmanae).     

循序间歇式废水生物除磷处理工艺微生物学特性的研究

考察了处理效果良好的循序间歇式废水生物除磷试验装置内活性污泥的微生物组成及其优势菌的磷代谢特性。试验结果表明,活性污泥混合液中的优势菌为假单胞菌属,其次是气单胞菌属和棒杆菌属,仅发现少量的不动杆菌属。当以乙酸钠为基质时,假单胞菌属于好氧培养过程中能明显除磷,其效果与厌氧/好氧培养过程的相近。另外,在好氧条件下,以乙酸钠为基质培养时,向培养基内滴加H₂SO₄可导致假单胞菌产生磷释放现象,滴加NaOH或NaHCO₃时,则发生过量摄磷现象,并且加NaOH所引起的磷摄取量较NaHCO₃的大。      

梯度负荷下果蔬垃圾厌氧消化性能及微生物群落结构的研究

为优化果蔬垃圾厌氧消化工艺,提高厌氧消化性能,本文通过逐级提高CSTR反应器进料负荷,研究不同负荷下的厌氧消化性能及相应的微生物群落结构变化规律.结果表明,随着进料负荷的增高,容积产气率、甲烷产气量、氨氮、碱度、TCOD、SCOD均逐渐增高,在最高负荷(负荷以VS计)2.50g·L-·1d-1时分别达到最大值:1.22L·L-1d-1,5.10L·d-1,1563.86mg·L-1,7572.23mg·L-1,13283.26mg·L-1,2075.03mg·L-1,甲烷含量及VFA分别稳定在52.46%～54.59%和(879.30±18.69)mg·L-1;同时利用PCR-DGGE技术系统分析了厌氧消化中细菌与古细菌的群落结构,测序结果表明,整个过程中拟杆菌(Bacteroidetes)、甲烷鬃菌(Methanosaeta)及甲烷螺菌(Methanospirillum)为优势微生物,随着负荷的提高,甲烷鬃毛菌(Methanosaeta)活性逐渐降低;聚类分析及主成分分析表明,低负荷条件下(1.50g·L-·1d-1、1.75g·L-·1d-1),微生物种类(细菌、古细菌)差别不明显,且基本处于同一阶段.     

Effective removal of antibiotic resistance genes and potential links with archaeal communities during vacuum-type composting and positive-pressure composting

As a major reservoir of antibiotics, animal manure contributes a lot to the augmented environmental pressure of antibiotic resistance genes (ARGs). This might be the first study to explore the effects of different ventilation types on the control of ARGs and to identify the relationships between archaeal communities and ARGs during the composting of dairy manure. Several ARGs were quantified via Real-time qPCR and microbial communities including bacteria and archaea were analyzed by High-throughput sequencing during vacuum-type composting (VTC) and positive-pressure composting (PPC). The total detected ARGs and class I integrase gene (intI1) under VTC were significantly lower than that under PPC during each stage of the composting (p < 0.001). The relative abundance of potential human pathogenic bacteria (HPB) which were identified based on sequencing information and correlation analysis decreased by 74.6% and 91.4% at the end of PPC and VTC, respectively. The composition of archaeal communities indicated that methane-producing archaea including Methanobrevibacter, Methanocorpusculum and Methanosphaera were dominant throughout the composting. Redundancy analysis suggested that Methanobrevibacter and Methanocorpusculum were positively correlated with all of the detected ARGs. Network analysis determined that the possible hosts of ARGs were different under VTC and PPC, and provided new sights about potential links between archaea and ARGs. Our results showed better performance of VTC in reducing ARGs and potential HPB and demonstrated that some archaea could also be influential hosts of ARGs, and caution the risks of archaea carrying ARGs.     

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.     

稻秸添加对两种水稻土产甲烷古菌及细菌的影响

稻秸还田对水稻土CH4排放有重要影响.本研究通过微宇宙厌氧培养,研究了两种水稻土[江西(JX)和广东雷州半岛(GD)]在稻秸(RS)添加条件下经相对较长时间的厌氧培养后产甲烷古菌及细菌菌群的响应.结果表明,不同RS添加量对JX水稻土产甲烷古菌群落结构有一定的影响,而对GD水稻土产甲烷古菌群落的影响不大. RS添加量与mcr A基因拷贝数之间存在显著正相关关系,JX水稻土的mcr A基因拷贝数变化对RS添加量的响应更敏感.相同RS添加条件下,JX水稻土mcr A基因拷贝数大于GD水稻土.相同稻秸量添加条件下两种水稻土的产甲烷古菌群落结构也有差异. JX水稻土共检测到的产甲烷古菌有Methanosarcinaceae、Methanocellaceae、Methanomicrobiaceae、Methanobacteriaceae以及未知菌群(494 bp). GD水稻土中仅有3大类产甲烷古菌,分别为Methanobacteriaceae、Methanosarcinaceae和Methanocellaceae.对2%RS处理培养第270 d的细菌菌群进行了测序,发现两种水稻土的细菌菌群明显不同,GD水稻土的细菌多样性高于JX水稻土,而其优势细菌数量(共有Longilinea、Acidobacteria/Gp6、Bellilinea及Thermosporothrix)低于JX水稻土(共有Bacillus、Desulfovirgula、Thermosporothrix、Acidobacteria/Gp1、Acidobacteria/Gp3及Ktedonobacter). RS作为底物促进产甲烷古菌的生长.不同类型的水稻土经相对较长时间的厌氧培养后其产甲烷古菌及细菌菌群结构也不同.     

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.     

反硝化聚磷菌快速富集、培养及其荧光原位杂交技术鉴别

以污水处理厂A2/O厌氧段污泥为种泥,采用膜生物反应器(membrane bio-reactor,MBR)对反硝化聚磷菌(denitrifyingphosphate-removal bacteria,DPB)进行快速富集及培养,并提供一种鉴别方法.试验中以乙酸钠为碳源,并在缺氧段投加一定浓度的硝酸盐,结果表明,在膜组件的高效截留作用下,经过厌氧-好氧和厌氧-缺氧2个阶段的富集培养,35 d内反硝化聚磷菌占全部聚磷菌(phosphate-accumulating organisms)的比例从24%上升到93%.此时系统的脱氮、除磷效率均可保持在90%以上.通过荧光原位杂交技术(fluorescence in situ hybridization,FISH)并结合常规测定手段对活性污泥进行鉴别,确定Pseudomonas sp.和Rhodocyclus sp.为主要的优势菌.     

Performance and microbial community of a membrane bioreactor system——Treating wastewater from ethanol fermentation of food waste

In this study, a lab-scale biological anaerobic/anaerobic/anoxic/membrane bioreactor(A_-~3MBR) was designed to treat wastewater from the ethanol fermentation of food waste,a promising way for the disposal of food waste and reclamation of resources. The 454 pyrosequencing technique was used to investigate the composition of the microbial community in the treatment system. The system yielded a stable effluent concentration of chemical oxygen demand(202 ± 23 mg/L), total nitrogen(62.1 ± 7.1 mg/L), ammonia(0.3 ±0.13 mg/L) and total phosphorus(8.3 ± 0.9 mg/L), and the reactors played different roles in specific pollutant removal. The exploration of the microbial community in the system revealed that:(1) the microbial diversity of anaerobic reactors A_1 and A_2, in which organic pollutants were massively degraded, was much higher than that in anoxic A_3 and aerobic MBR;(2) although the community composition in each reactor was quite different, bacteria assigned to the classes Clostridia, Bacteroidia, and Synergistia were important and common microorganisms for organic pollutant degradation in the anaerobic units, and bacteria from Alphaproteobacteria and Betaproteobacteria were the dominant microbial population in A_3 and MBR;(3) the taxon identification indicated that Arcobacter in the anaerobic reactors and Thauera in the anoxic reactor were two representative genera in the biological process. Our results proved that the biological A_-~3MBR process is an alternative technique for treating wastewater from food waste.     

厌氧折流板反应器(ABR)中微生物种群演替特征

采用SEM,FISH和PCR-DGGE技术对ABR(厌氧折流板反应器)各隔室中厌氧颗粒污泥进行分析,考察微生物形态、真细菌数量及种群结构演替过程,并对优势菌种进行了系统发育分析. 结果表明:ABR反应器颗粒污泥微生物中杆菌占优势,其中前端的微生物生长较好,活性高;沿反应器流程方向,各隔室微生物总量逐渐降低,真细菌相对丰度随之递减,其中1#隔室真细菌相对丰度最高,为65.9%,而5#隔室只有27.2%. 此外,ABR反应器前端以真细菌为主,而后端隔室古细菌含量升高,微生物种群随流程发生显著演替,但5个隔室间真细菌的Shannon-Wiener多样性指数没有显著性变化. UPMGA聚类分析表明,1#隔室与2#隔室的微生物群落相似性为77%,4#隔室与5#隔室的相似性为85%,3#隔室与其他隔室的相似性均较低,表明ABR反应器前端以发酵产酸作用为主,后端以产甲烷作用为主,ABR反应器具有明显的分阶段多相工艺特点.      

Functional relationship between ammonia-oxidizing bacteria and ammonia-oxidizing archaea populations in the secondary treatment system of a full-scale municipal wastewater treatment plant

The abundance of ammonia-oxidizing bacteria and archaea and their amoA genes from the aerobic activated sludge tanks, recycled sludge and anaerobic digesters of a full-scale wastewater treatment plant (WWTP) was determined. Polymerase chain reaction and denaturing gradient gel electrophoresis were used to generate diversity profiles, which showed that each population had a consistent profile although the abundance of individual members varied. In the aerobic tanks, the ammonia-oxidizing bacterial (AOB) population was more than 350 times more abundant than the ammonia-oxidizing archaeal (AOA) population, however in the digesters, the AOA population was more than 10 times more abundant. Measuring the activity of the amoA gene expression of the two populations using RT-PCR also showed that the AOA amoA gene was more active in the digesters than in the activated sludge tanks. Using batch reactors and ddPCR, amoA activity could be measured and it was found that when the AOB amoA activity was inhibited in the anoxic reactors, the expression of the AOA amoA gene increased fourfold. This suggests that these two populations may have a cooperative relationship for the oxidation of ammonia.