Comparative study of microbial community structure in different filter media of constructed wetland

Comparisons of microbial community structure, in eight filter media of zeolites, anthracite, shale, vermiculite, ceramic filter media, gravel, steel slag and bio-ceramic, were undertaken by analyzing the phospholipid fatty acid (PLFA) composition. A total of 20 fatty acids in the range of C_(11) to C_(20) were determined but only 13 PLFAs were detected in steel slag. They consist of saturated fatty acids, branched fatty acids, monounsaturated fatty acids, and polyunsaturated fatty acids. The variation of fatty acids was revealed in the relative proportions of these fatty acids in different media. The aerobic prokaryotes were the predominant group in all media. The PLFA composition showed significant differences among the eight different media by Tukey's honestly test. It was found that steel slag was significantly different in the microbial community as compared to other filter media, probably due to its alkaline effluent. Steel slag alone is probably not a good choice of substratum in constructed wetlands. The principle components analysis (PCA) showed that zeolites, bio-ceramic, shale and vermiculite had a similar microbial community structure while steel slag and ceramic filter media were distinct from other media.     

利用磷脂脂肪酸表征人工湿地微生物群落结构

通过磷脂脂肪酸(PLFAs)分析,研究了复合垂直流人工湿地基质剖面的微生物群落结构及其分布特征.结果表明,湿地PLFAs组成以饱和脂肪酸、支链脂肪酸和单不饱和脂肪酸为主,多不饱和脂肪酸与环丙烷脂肪酸含量较少.特征脂肪酸的比值呈垂直分布.基质中好氧原核微生物为优势类群,其次为革兰氏阳性细菌及其他厌氧细菌,真核微生物所占比例最低.统计结果显示,好氧原核微生物的特征PLFAs相对含量在上行池表层最高;革兰氏阳性细菌、SRB及其他厌氧细菌的特征PLFAs相对含量在下行池中层显著高于其他位点,指示该区域为湿地系统主要的兼性厌氧功能区.     

湿地型微生物燃料电池运行实验研究

为了探究湿地植物、电极间距对湿地型生物燃料产电及污水净化效能的影响,本文选用鸢尾、绿萝、富贵竹和观音竹4种湿地植物构建湿地型生物燃料电池与相同结构无种植湿地植物的燃料电池进行对照.结果表明,有植物的微生物燃料电池的产电性能优于无植物的微生物燃料电池;4种植物微生物燃料电池功率密度由高到低的顺序依次为:鸢尾绿萝富贵竹观音竹.相同绿萝植物的湿地型微生物燃料电池在不同的电极间距下的产电性能不同,间距越小输出电压和功率密度越大.当电极距离为10 cm时,最大功率密度达2.55 W·m~(-3),比20 cm和30 cm分别高约30%和50%.4种湿地植物的湿地型微生物燃料电池对COD的去除率均达80%以上,COD去除效率由高到低的顺序依次为:鸢尾富贵竹观音竹绿萝.     

辽河保护区人工湿地微生物群落结构及分布规律

利用高通量测序技术,研究了辽河保护区盘锦段3种类型人工湿地(支流汇入口湿地、坑塘湿地、牛轭湖湿地)底泥和芦苇根际微生物群落结构与分布规律。结果表明: 3种类型人工湿地底泥微生物多样性、丰度和操作分类单元(optical transform units,OTUs)数目均大于芦苇根际微生物,且底泥和芦苇根际微生物多样性受季节变化影响,秋季高于春季;不同类型人工湿地微生物群落结构及分布大致相似,变形菌门(Proteobacteria)为相对丰度最高的优势种,以变形菌门为主的革兰氏阴性菌在人工湿地净化过程中起重要作用;不同类型人工湿地变形菌纲及其相对丰度不同,支流汇入口湿地以α-变形菌纲(Alphaproteobacteria)、δ-变形菌纲(Deltaproteobacteria)和γ-变形菌纲(Gammaproteobacteria)为主;坑塘湿地以γ-变形菌纲和α-变形菌纲为主;牛轭湖湿地中不同纲的微生物分布较均匀。     

Evaluation of the e ectiveness of horizontal subsurface flow constructed wetlands for di erent media

Two media bed (gravel and Filtralite NR) were tested in a mesocosm to evaluate the removal of organic matter (as chemical oxygen demand (COD)), ammonia (NH4-N), nitrite, nitrate and solid matter (as total suspended solids (TSS)) for a synthetic wastewater (acetate-based) and a domestic wastewater. The use of Filtralite allowed average removal rates (6–16.8 g COD/(m2 day), 0.8–1.1 g NH4-N/(m2 day) and 3.1 g TSS/(m2 day)) and removal e ciencies (65%–93%, 57%–85% and 78% for COD, NH4-N and TSS, respectively), higher than that observed in the experiments with gravel. The applied loads of COD, ammonia, nitrate and TSS seem to influence the respective removal rates but only for the treatment of domestic wastewater with higher correlation coe cients for Filtralite. Regardless the type of media bed and the type of wastewater, nitrate was completely removed for nitrogen loading rates up to 1.3 g NO3-N/(m2 day). There was no evidence of the influence of nitrate loads on the removal of organic matter.     

构建湿地中试系统基质剖面微生物活性的研究

对稳定运行的复合垂直流构建湿地中试系统基质进行分层采样,测定微生物数量,生物量氮,酶活性,硝化作用,反硝化作用以及呼吸作用等各项指标.结果表明,上行流和下行流池0～10cm的基质层中好氧微生物数量高出30～55cm的层面1～2个数量级;反硝化细菌除下行流池0～10cm的层面外其他基质层数量都达到10⁷个/g干土以上.0～20cm的基质层中脲酶活性明显高于30～55cm的层面;而0～10cm的层面中脱氢酶活性明显高出10～55cm的层面2～5倍.硝化反硝化作用广泛存在于基质中,硝化作用表现出一定的空间规律,0～10cm的基质硝化作用强度要比40～55cm层面的高出20%;反硝化作用的空间差别很小,其作用率最低也达到85.5%.     

Characteristics of the microbial communities in the integrated vertical-flow constructed wetlands

Microorganisms play an important role in removing pollutants from constructed wetlands. We investigated the microbial characteristics in a novel integrated vertical-flow constructed wetland (IVCW), which has been in operation in Wuhan, China since 1998. We used phospholipid fatty acid (PLFA) and amoA gene to analyze the structure and diversity of the microbial community within the IVCW. PLFA results suggested that the amount of bacterial PLFA was significantly higher than that of fungal PLFA, but the total microbial biomass represented by PLFA index was low in the system. Microbial spatial distribution showed significantly higher bacterial (both G⁺ and G⁻) and fungal biomass in the surface than in the subsurface layers. The ratios of monounsaturated to branched PLFA demonstrated that an anaerobic layer sandwiched by two aerobic layers existed in the IVCW, consistent with the redox potential results. Analysis of the amoA revealed the presence of Nitrosomonas-like sequences in the surface substrate of the downflow chamber and apparent diversities of ammonia-oxidizing bacteria in the system. These results suggest that microorganisms, despite their relatively low biomass, have inhabited the IVCW, and the results will offer some valuable information on microbe to system designers and managers.     

不同结构好氧/厌氧潜流人工湿地微生物群落代谢特性

4种不同结构的好氧/厌氧多级串联潜流人工湿地对COD和氮的去除效果不同.为了探究基质微生物群落代谢特征及其与水质净化效果的关系,对利用Biolog微平板得到的AWCD值(平均每孔光密度值)进行碳源分类、主成分分析以及聚类分析.结果表明,微生物对于糖类及其衍生物和氨基酸及其衍生物的利用水平明显高于对脂肪酸及脂类和代谢中间产物及次生代谢物的利用水平;OBAAO(好氧-缓冲-厌氧-缺氧-好氧)曝气组中采样点3.3(厌氧采样点)微生物对4类碳源的利用水平都最低,与其他各采样点具有显著性差异(p<0.05),96hAWCD值主成分分析和聚类分析的结果表明,OBAAO曝气组采样点3.3中的微生物群落和其他3个进行厌氧反应的采样点中的微生物群落发生了较大的差异,OBAAO曝气组延长厌氧段长度为微生物提供厌氧环境,而由于缺少碳源作为能量,使得微生物的活性受到抑制;OBAAO曝气多点进水组中的采样点4.3(厌氧采样点)微生物在4类碳源的利用水平上都具有一定的优势性,OBAAO曝气多点进水组补充进水和延长厌氧段长度两个措施有效地促进了微生物的反硝化作用的强度,提高了水平潜流人工湿地的氮去除率.     

季节性冻融对扎龙湿地土壤微生物群落结构和胞外酶活性的影响

选取扎龙湿地芦苇沼泽为研究对象,采用磷脂脂肪酸法和微孔板法研究了季节性冻融期土壤微生物群落结构和胞外酶活性的动态变化规律.结果表明,微生物总磷脂脂肪酸（PLFA）含量和细菌PLFA含量均呈先降低后升高趋势,而真菌PLFA含量呈先升高后降低趋势.冻融初期至融冻初期,β-葡萄糖苷酶显著降低了71.84%,β-N-乙酰氨基葡萄糖苷酶表现为先升高后降低趋势,酸性磷酸酶在融冻初期最高、冻融初期最低,过氧化物酶和多酚氧化酶峰值均出现在融冻后期.主成分分析表明,季节性冻融显著改变了土壤微生物群落结构.冗余分析发现,土壤含水率与总PLFA、细菌、革兰氏阳性菌、革兰氏阴性菌、放线菌、过氧化物酶和多酚氧化酶呈显著正相关（p<0.05）;土壤全磷、速效磷、速效氮与真菌和β-N-乙酰氨基葡萄糖苷酶呈显著正相关;β-葡萄糖苷酶主要受土壤碳氮比和有机质影响,而土壤微生物与胞外酶活性在不同冻融时期具有显著相关性（p<0.05）.因此,探究全球变化背景下土壤微生物和胞外酶对季节性冻融过程的响应机制,可为深入解析扎龙湿地土壤质量演变特征及湿地生态系统功能的恢复与保护提供科学依据.     

垂直潜流人工湿地低温净化效果及其与微生物作用关系

人工湿地净化过程受基质、微生物及植物的多重作用,但有关湿地系统水质净化效果与微生物之间的关联及响应机制的认识较为匮乏.本文针对垂直潜流人工湿地,分析了其低温条件下水质净化效果,通过16S rDNA扩增子测序技术,在OTU水平上利用微生物丰度聚类热图和基于Bray-Curtis距离的NMDS分析,揭示了湿地基质微生物群落结构的时空变化及其与主要水质指标之间的相互作用关系.结果表明,湿地对TN、COD的去除效果较好,但对TP的去除效果不佳.基质微生物群落时空变化显著,与湿地净化效果密切相关,革兰氏阴性菌门类众多和假单胞菌属相对丰度较低,分别是TN去除效果较好和TP去除效果不佳的重要原因.COD、DO是影响微生物群落的主要因子,而氮含量在一定程度上影响脱氮微生物的活跃度.基质微生物在湿地运行2周后,对污染物去除的介导作用稳定,建议设立这一关键时间点,并辅以菌群、碳源投入等措施,提升湿地的净化效果.     

不同填料甲苯生物滤塔中微生物群落结构与代谢功能解析

对填料为珍珠岩、玻璃珠和聚苯乙烯泡沫(EPS)的甲苯生物滤塔中微生物群落的结构和代谢功能进行了分析.结果表明,在稳定运行阶段,生物滤塔的微生物群落结构相似且与接种物存在较大差异.3种填料的生物滤塔中在门水平上的优势菌均为变形菌门Proteobacteria(36.1%~67.5%)、放线菌门Actinobacteria(4.3%~43.4%)、厚壁菌门Firmicutes(3.0%~16.4%)和拟杆菌门Bacteroidetes(1.9%~10.2%);属水平上,相同的优势属为红球菌属Rhodococcus(2.4%~38.5%)和假单胞菌属Pseudomonas(5.9%~25.3%),在珍珠岩生物滤塔中相对丰度更高的属为戴沃斯氏菌属Devosia、戈登氏菌属Gordonia和根瘤菌科某属Rhizobiales_E,在玻璃珠和EPS生物滤塔中相对丰度更高的属为着色菌科某属Chromatiaceae_A和Fluviicola.不同填料的生物滤塔中微生物群落整体碳源代谢能力相似且相比接种物均显著下降,微生物群落结构与代谢呈现一致性,多样性水平均为珍珠岩EPS玻璃珠.与接种物相比,不同填料的生物滤塔中微生物结构物种多样性都降低,代谢多样性都升高.     

Soil enzymatic activities and microbial community structure with di erent application rates of Cd and Pb

This study focused on the changes of soil microbial diversity and potential inhibitory effects of heavy metals on soil enzymatic activities at different application rates of Cd andor Pb. The soil used for experiments was collected from Beijing and classified as endoaquepts. Pots containing 500 g of the soil with different Cd andor Pb application rates were incubated for a period of 0, 2, 9, 12 weeks in a glasshouse and the soil samples were analyzed for individual enzymes, including catalase, alkaline phosphatase and dehydrogenase, and the changes of microbial community structure. Results showed that heavy metals slightly inhibited the enzymatic activities in all the samples spiked with heavy metals. The extent of inhibition increased significantly with increasing level of heavy metals, and varied with the incubation periods. The soil bacterial community structure, as determined by polymerase chain reaction-denaturing gradient gel electrophoresis techniques, was different in the contaminated samples as compared to the control. The highest community change was observed in the samples amended with high level of Cd. Positive correlations were observed among the three enzymatic activities, but negative correlations were found between the amounts of the heavy metals and the enzymatic activities.     

人工湿地中除磷填料的筛选及其除磷能力

针对猪场废水水质特点,对4种填料的理化特性进行了分析,并考察了4种填料的除磷能力.结果表明,废砖块和沸石的表面微结构更有利于生物膜的生长,而海蛎壳与废砖块则具有较高的除磷潜能;4种填料对磷素的理论饱和吸附量依次为海砺壳>废砖块>火山岩>沸石,海蛎壳对磷素的吸附主要为化学吸附,而废砖块、火山岩和沸石对磷素的吸附主要为物理吸附,其中废砖块对磷素的吸附过程中存在化学吸附作用;在动态吸附试验中,海蛎壳对磷素的去除效果最好,其次为废砖块、火山岩和沸石,4种填料去除磷素的主要途径由其化学成分和化学形态决定.由试验结果可知,海蛎壳是人工湿地处理猪场废水时较为理想的除磷填料.     

人工湿地污水处理系统春季空气微生物群落结构分析?

通过构建16S/18S rDNA基因文库,分析自由表面流人工湿地污水处理系统春季空气细菌和空气真菌群落结构特征.结果表明,空气细菌分布在变形菌门(Proteobacteria)、放线菌门(Actinobacteria)、浮霉菌门(Planctomycetes)、蓝藻门(Cyanophyta)、绿弯菌门(Chloroflexi)、拟杆菌门(Bacteroidetes)和厚壁菌门(Firmicutes),主要为β-变形菌纲(71.04%)、γ-变形菌纲(12.03%)、α-变形菌纲(3.83%)、蓝藻纲(4.38%)、芽孢杆菌纲(3.28%)和鞘脂杆菌纲(2.19%),优势菌属是马赛菌属(Massilia 66.66%)、假单胞菌属(Pseudomonas 4.37%)、蓝丝细菌属(Cyanothece 3.83%)和沙雷氏菌属(Serratia 3.28%).空气真菌主要类群为座囊菌纲(Dothideomycetes 61.18%),其次是接合菌纲(Zygomycetes 16.47%)、盘菌纲(Discomycetes 14.12%),优势菌属是核腔菌属(Pyrenophora 48.31%)、被孢霉属(Mortierella 15.7%)、缘刺盘菌属(Cheilymenia 12.4%)、Boothiomyces (4.5%).人工湿地空气微生物中未检测出大肠杆菌(Escherichia coli)、沙门氏菌(Salmonella spp.)和产气荚膜梭菌(Clostridium perfringens),但存在粘质沙雷氏菌(S. marcescens)、恶臭假单胞菌(P. putida)、表皮葡萄球菌(Staphylococcus epidermidis)等致病菌或条件致病菌.     

高通量测序法表征潜流人工湿地中不同植物根际细菌群落特征

为了研究人工湿地中植物根际、污水水质和深度等对细菌群落结构特征分布的影响,利用高通量测序技术,对人工湿地中芦苇(Phragmites communis)、香蒲(Typha orientalis Presl)2种植物根际3个不同深度细菌群落特征进行了研究.细菌群落丰富度和多样性研究结果表明,芦苇根际细菌群落丰富度和多样性均大于香蒲根际,细菌在芦苇根际周围可以更好地生存;同一植物根际细菌的丰富度和多样性随着深度的增加逐渐减少.相似度和差异性分析结果表明,相同植物根际细菌群落结构相似度较高,而不同根际群落结构有一定的差别.优势细菌菌群分析结果发现,细菌群落在门类水平上达到13门以上,优势细菌种群均以变形菌门、酸杆菌门、绿弯菌门、厚壁菌门为主,相对丰度约为55%~78%;纲类水平也达到20纲以上,主要有α-变形杆菌纲、β-变形菌、δ-变形菌纲、γ-变形菌纲、芽孢杆菌纲、酸杆菌纲、相对丰度达到50%以上,植物根际富集的主要纲类细菌是β-变形菌纲.影响细菌群落结构丰富度和多样性的主要环境因素是营养物浓度、植物、采样深度和温度.     

生物膜脱氮滤池的微生物群落结构特性

为探究生物膜脱氮滤池脱氮效能差异的微生物因素,设置组1(包括接种污泥、稳定期滤料、反冲洗后滤料微生物变化)和组2(包括反冲洗前滤料、反冲洗后不同时间段滤料微生物变化)两组实验,通过高通量测序,研究不同阶段微生物群落结构、丰度和多样性的分布情况.结果表明:接种污泥与滤料表面微生物群落结构和多样性在门水平下差异不大,但在属水平下差异显著,而相对丰度在两种分类水平下始终差异显著.反冲洗后0～4 h滤料表面微生物的OTU数目、Shannon指数、Chao1指数变化,在滤池30 cm处先升后降,而在60 cm处曲折上升,也即二者的微生物丰度和多样性变化趋势.反冲洗前后Proteobacteria始终占主导地位,相对丰度为79%～90%,Proteobacteria中Betaproteobacteria占主导优势,反冲洗后Betaproteobacteria数量相对减少,后逐渐恢复.总反硝化优势菌属相对丰度在反冲洗1 h后30 cm处由78%左右下降到70%左右,60 cm处由68%左右下降到64%左右,此时出水总氮达到最高值7.7 mg·L~(-1),之后总反硝化优势菌属及出水TN浓度逐渐恢复至正常水平,这种消长变化表明滤池脱氮效果与总反硝化优势菌属相对丰度密切相关,滤池运行状态的改变使得反硝化优势菌属的群落结构差异显著.     

Effect of nutrient level on phytoplankton community structure in di erent water bodies

Increasing levels of pollution within water bodies can cause eutrophication and an associated rapid growth in and reproduction of phytoplankton. Although most frequently occurring in bodies of water such as lakes and dams, in recent years an increasing number of river systems in China have suffered serious algal blooms. The community structure of phytoplankton may differ, however, dependent on the hydrodynamic conditions and nutrient levels within the water body. The field investigation results obtained fro...     

磺胺嘧啶胁迫下作物根际微生物群落结构响应

为了研究在磺胺类兽药胁迫下,两种作物在不同生长期根际微生物群落结构的响应,采用室内根际箱培养实验,测定玉米、小麦根际微界面土壤磷脂脂肪酸的量,研究在磺胺嘧啶（SD）胁迫下,不同作物根际土壤微界面微生物群落结构的空间变化.结果表明,SD对根际微生物活性的有抑制作用,且强度随浓度增加而增强.同一浓度SD作用下,根际微界面微生物生物量不同,在实验中,根际3mm和根室的微生物生物量最大,且二者之间差异不显著.不同微生物对根际效应敏感程度不同,细菌、革兰氏阳性菌、革兰氏阴性菌、蓝细菌和硫酸盐还原菌根际效应明显.在SD胁迫下,根际不同微界面土壤微生物群落结构变化明显.细菌、G⁺、G^-、放线菌的生物量,随SD浓度升高而下降,主要表现为抑制效应,而对真菌生物量则表现为激活效应,生物量增加. 小麦根际土壤真菌：细菌（F/B）随SD浓度的升高,比值增大,在高浓度（5mg/kg）胁迫下,F/B比值最大（0.74）,与对照差异显著（P < 0.05）,说明土壤污染修复能力增强,而sat/mono比值在高浓度SD胁迫下降低,说明微生物群落结构向有利于SD降解的方向转化.根际效应有助于SD的降解,在1mg/kg SD作用下,小麦根际土壤降解率为7.01%,而非根际土壤降解率仅为2.49%,不同作物表现出的根际效应强弱不同,玉米根际效应强于小麦.     

石臼漾人工湿地冬夏季全程氨氧化微生物的分布、群落结构及其影响因素研究

湿地作为水体与陆地之间的过渡地带,具有独特的生态环境特性,是氮循环反应的关键区域.研究湿地生态系统中的全程氨氧化过程（complete ammonia oxidation,comammox）,解析该区域中comammox细菌的分布与群落结构特征,补充了此前该新型氨氧化微生物在湿地生态系统中分布特征的研究空白,对于完善comammox细菌在各种不同生态系统中分布情况的研究具有重要意义.本研究在石臼漾人工湿地中于冬夏两季分别采集了大沟中心、大沟边缘、小沟中心和小沟边缘的表层沉积物样品,利用PCR、荧光定量PCR和amoA功能基因高通量测序等方法,解析人工湿地中comammox细菌的时空分布与群落结构特征.主要研究结果如下：①所有样品中均检测到了comammox细菌,其丰度为1.77×10⁵~4.07×10⁷ copies·g^-1.冬季,comammox细菌丰度在大沟中高于氨氧化细菌（ammonia oxidizing bacteria,AOB）和氨氧化古菌（ammonia oxidizing archaea,AOA）,在小沟中高于AOB,但低于AOA;夏季,comammox细菌丰度在所有样点中均高于AOB和AOA.②冬夏两季样品中,comammox细菌丰度与底物NH₄⁺-N浓度均呈负相关关系,且夏季样品中呈显著性负相关.③主坐标分析（Principle Coordinate Analysis,PCoA）与多样性分析结果表明,comammox细菌群落结构具有空间异质性,且冬季物种多样性高于夏季.     

轮休措施对堵塞型垂直潜流人工湿地的影响

采用不可滤物质的定量化分析方法研究了采取轮休措施对人工湿地堵塞程度的改善效果,同时分析了轮休措施对垂直潜流人工湿地污染物去除效果的改变和植物生长的影响.结果表明,轮休措施对解决人工湿地的堵塞有明显地改善,采取轮休措施后单位基质中的不可滤物质含量较轮休前有所降低,其中不可滤有机物减少量十分明显,不可滤无机物量在短期和长时间轮休前后分别表现出基本稳定和较大幅度降低的特性;短期轮休后COD、TP和TN的去除率都较轮休前有所降低,氨氮的去除率在人工湿地轮休后较轮休前高;应结合人工湿地的基质、植物种类调整运行工况,进行不同时间长度和方式的轮休,以确保植物生长不受该措施的影响.