湿地法处理生活污水对仙桃湿地地表水微生物群落结构的影响

【背景】随着中小城市经济的高速发展和人们生活水平的提高,生活污水排放量也日益增大,致使水源水质污染呈恶化趋势。【目的】了解湿地法处理生活污水对湿地地表水微生物群落多样性变化的影响。【方法】对人工湿地(仙桃)参照点(-2 000)、200、400、600、3 000、5 000 m水平方向6点地表水进行理化检测,再通过高通量测序对微生物群落结构进行解析,并探讨微生物群落结构变化与生活污水的污染效应。【结果】当湿地法处理的生活污水到达5 000 m时,化学需氧量、氨态氮、总氮和总磷均达《城镇污水处理厂污染物排放标准》一级标准,且理化指标之间均呈极显著正相关。样品在门水平上的优势物种分别为变形菌门(Proteobacteria)、拟杆菌门(Bacteroidetes)、放线菌门(Actinobacteria)、蓝藻门(Cyanobacteria)、疣微菌门(Verrucomicrobia)、厚壁菌门(Firmicutes)、酸杆菌门(Acidobacteria)和绿弯菌门(Chloroflexi)。变形菌门(Proteobacteria)可用来作为排放生活污水污染物的指示性微生物,放线菌门(Actinobacteria)、蓝藻门(Cyanobacteria)、疣微菌门(Verrucomicrobia)和绿弯菌门(Chloroflexi)可用来作为排放生活污水净化的指示性微生物。【结论】首次比较分析了湿地法处理仙桃生活污水地表水环境中微生物群落多样性变化的情况,可使人们从微生物层面更加明确地了解人工湿地运作机理及地表水环境的污染情况。     

不同类型人工湿地微生物群落的研究进展

微生物是人工湿地不可缺少的成员,对湿地生态系统中物质转化、能量流动起着重要作用.本文从人工湿地微生物群落的研究方法、微生物群落结构与组成、微生物群落调节作用与环境因素的关系等方面,综述了人工湿地微生物的研究进展.各种新颖的分子生物学方法已经成为研究人工湿地的微生物多样性的有力工具,其中最常见的是变性梯度凝胶电泳(DGGE)和寡核苷酸荧光探针原位杂交(FISH);人工湿地微生物群落的调节作用主要取决于湿地的水文条件、废水的特点(包括组成成分,污染物的特点和利用性)、湿地的过滤材料或土壤类型、植物和各种环境因素;不同人工湿地类型的微生物群落组成,从多到少依次是变形菌、噬纤维菌.黄杆菌菌群、放线菌和厚壁菌.如何进一步加深对氮循环相关微生物多样性的研究,提高废水中氮的去除效率依然是未来人工湿地技术需要解决的重要问题之一.     

水平潜流人工湿地堵塞初期微生物群落结构变化

为了寻找合适的湿地堵塞监测指标, 文章通过运行4组湿地装置来模拟水平潜流人工湿地堵塞的过程, 并研究人工湿地中微生物群落在湿地堵塞初期的响应。在人工湿地反应器运行过程中, 定期监测基质过滤速率与人工湿地处理性能, 包括化学需氧量(COD)、总氮(TN)和总磷(TP)去除效率, 同时应用高通量测序技术测定并分析湿地反应器中微生物群落结构组成与多样性。结果表明: 随着堵塞的形成, 在4组人工湿地中, 基质过滤速率和TP的去除效率持续下降, 而COD和TN的去除效率分别为50%—85%和10%—20%; 16S rDNA测序结果表明堵塞会降低细菌群落的丰富度和多样性; 人工湿地系统中的优势菌门主要是变形菌门(Proteobacteria)、厚壁菌门(Firmicutes)和绿弯菌门(Chloroflexi)等, 且当湿地进入堵塞初期后, 变形菌门丰度明显增加, 具体表现为其门下的反硝化细菌属(Thauera、Zoogloea和Rhizobium) 丰度显著增加, 此外厚壁菌门和绿弯菌门的有机物降解菌属(Clostridium sensu stricto、Gracilibacter和Levilinea)的丰度也明显增加, 这表明上述菌属有望成为人工湿地系统堵塞初期的预警指示菌。     

人工湿地系统微生物去除污染物的研究进展

人工湿地系统是模拟自然湿地,通过植物、土壤、微生物的相互作用去除污染物,从而实现净化污水、美化环境的功能,是一种经济环保且高效的污水处理技术。微生物作为人工湿地生态系统的重要组成部分,承担着对污水、废水中污染物的吸收和分解。本文从微生物种群分布特征和影响微生物生态分布及去污能力的三个因素,温度和营养物质、基质方面,总结归纳了人工湿地微生物去除污染物的研究进展。     

造纸废水活性污泥的微生物群落结构及多样性分析

为探究造纸废水活性污泥中微生物群落结构多样性以及对造纸废水处理效果的影响，利用Illumina MiSeq 高通量测序方法，分析在处理造纸废水过程中，同一运行阶段两个并联氧化沟内活性污泥的微生物群落与多样性组成。结果表明，系统中处理造纸废水的活性污泥在同一废水条件下微生物群落结构总体稳定，优势细菌为绿弯菌门(Chloroflexi)、拟杆菌门（Bacteroidota）、变形菌门（Proteobacteria）、Myxococcota、放线菌门(Actinobacteria）、厚壁菌门(Firmicutes）等。最重要的优势细菌类群为Chloroflexi，相对丰度占比为47.67%~48.22%，远远高于其他废水中Chloroflexi的占比，其中厌氧绳菌纲（Anaerolineae）是其主要成员，占比84.39%~88.34%，可针对性地去除造纸废水中的污染物。造纸废水活性污泥样品中存在大量特殊功能菌群，其在废水中污染物尤其是木质素的去除中发挥着重要作用。     

东太湖地区湿地水质与浮游细菌群落结构关联性研究

[目的]考察东太湖地区湿地水质与浮游细菌群落结构之间的关联,探索对湿地水质具有指示作用的微生物。[方法]应用高通量测序研究浮游细菌群落结构;通过相关性分析考察了浮游细菌群落多样性与水质的关联,并运用LEf Se分析定位不同水质水样的标志微生物。[结果]东太湖湿地浮游细菌主要分布于放线菌门(相对丰度34.78%)、变形菌门(26.30%)及蓝细菌门(19.28%)等7个门;水质指标综合得分与浮游细菌群落香农-威纳指数和辛普森指数分别显著相关(r=-0.597,p0.05;r=0.694,p0.01);蓝细菌门与伯克氏菌科的相对丰度在不同水质水样中差异最为显著(LDA得分4.0)。[结论]东太湖地区,较高的浮游细菌多样性表征较好的湿地水质;蓝细菌相对丰度较高(27.80%)指示湿地水质较差,而伯克氏菌相对丰度较高(5.48%)指示湿地水质相对较好。     

山西左云县采煤区人工湿地冬季沉积物细菌群落多样性

【背景】人工湿地处理煤炭矿井废水效果显著,而关于人工湿地沉积物的细菌群落结构和特征研究较少。【目的】阐明人工湿地净化采煤区废水的效应和细菌群落结构及特征。【方法】对山西省左云县采煤区人工湿地不同采样点水质进行监测,利用高通量测序技术对人工湿地沉积物中细菌16S rRNA基因V3-V4区进行测序,分析沉积物中细菌群落组成。【结果】采煤区废水流经人工湿地后水质得到有效改善,其中生化需氧量(Biochemical oxygen demand,BOD5)、化学需氧量(Chemical oxygen demand,CODCr)、总氮(Total nitrogen,TN)、总磷(Total phosphorus,TP)去除率分别达到76.2%、93.4%、73.4%和99.3%;测序共获得2 832个操作分类单元(Operational taxonomic units,OTUs),共753个属,分布于51个门的150个纲;4个采样点中优势菌门为变形菌门(Proteobacteria,50%-64.7%)和拟杆菌门(Bacteroidetes,15.8%-21.2%),变形菌门中优势菌纲为β-变形菌纲和γ-变形菌纲,其中M1、M2、M3中富含氨氮氧化细菌的β-变形菌纲和γ-变形菌纲丰度最高;M4沉积物细菌OTU最多,多样性最高;沉积物特有优势菌属Sulfurimonas、Sulfuricurvum的相对丰度与NH4+-N的含量显著负相关,Azoarcus菌属相对丰度与TN存在显著正相关,Novosphingobium菌属相对丰度与TP含量存在显著正相关。【结论】煤矿废水流经人工湿地可得到有效的净化,细菌多样性在人工湿地生态功能中起到了决定作用。     

污水处理过程中微生物群落多样性及其对环境因子响应的研究进展

污水生物处理系统的性能和稳定性与微生物群落结构和动态密切相关。通过深入了解活性污泥中微生物群落结构及其影响因素,有助于提高污水厂污染物的去除效果。在不同污水活性污泥处理系统中细菌群落主要以变形菌、绿弯菌、放线菌、厚壁菌和拟杆菌为功能菌群;活性污泥中寄居的大多数真菌来自于子囊菌门,还有少量担子菌门;古菌以产甲烷菌为主;而病毒中分布最广的噬菌体和致病性病毒是最主要的关注点。本文通过对相关文献分析及总结,综述了进水组成、不同处理工艺、参数(理化参数和运行参数)、地理位置和气候条件等环境因子对活性污泥中细菌、真菌、古菌以及病毒群落组成的影响,尽可能全面地介绍污水厂微生物群落多样性及其对环境因子的响应。同时,对未来研究方向进行探讨,以期能够为活性污泥中功能微生物的应用及调控提供理论和应用基础。     

东太湖地区湿地水质与浮游细菌群落结构关联性研究北大核心

[目的]考察东太湖地区湿地水质与浮游细菌群落结构之间的关联,探索对湿地水质具有指示作用的微生物。[方法]应用高通量测序研究浮游细菌群落结构;通过相关性分析考察了浮游细菌群落多样性与水质的关联,并运用LEf Se分析定位不同水质水样的标志微生物。[结果]东太湖湿地浮游细菌主要分布于放线菌门(相对丰度34.78%)、变形菌门(26.30%)及蓝细菌门(19.28%)等7个门;水质指标综合得分与浮游细菌群落香农-威纳指数和辛普森指数分别显著相关(r=-0.597,p<0.05;r=0.694,p<0.01);蓝细菌门与伯克氏菌科的相对丰度在不同水质水样中差异最为显著(LDA得分>4.0)。[结论]东太湖地区,较高的浮游细菌多样性表征较好的湿地水质;蓝细菌相对丰度较高(27.80%)指示湿地水质较差,而伯克氏菌相对丰度较高(5.48%)指示湿地水质相对较好。     

复合垂直流人工湿地系统中不同植物根际微生物群落结构

为了研究复合垂直流人工湿地在夏季和秋季运行期间湿地中不同植物根际微生物群落的结构特征,采用PCR-DGGE技术和基因测序技术,对复合垂直流人工湿地系统中梭鱼草(Pontederia cordata)、美人蕉(Canna indica)、再力花(Thalia dealbata)3种植物根际微生物群落的结构特征和组成多样性进行分析。结果表明,3种植物根际微生物的种类和数量各不相同。人工湿地下行池中梭鱼草、美人蕉、再力花根际微生物的Shannon指数分别为2.91、2.96和2.90,说明美人蕉根际能更好地富集周围的微生物,提高微生物群落的多样性。通过比较各种植物根际微生物的相似度,发现根际微生物群落结构存在一定的季节差异;美人蕉和再力花根际微生物群落结构相似度较高(65%以上)。微生物群落组成多样性的结果显示,复合垂直流人工湿地系统中3种植物在实验期间根际微生物群落分属8个纲:α-变形菌纲(14.37%)、β-变形菌纲(38.13%)、γ-变形菌纲(6.87%)、ε-变形菌纲(16.87%)、δ-变形菌纲(6.87%)、放线菌纲(8.76%)、拟杆菌纲(4.38%)、硝化螺旋菌纲(3.75%)。β-变形菌纲为该人工湿地系统试验期间不同植物根际富集的主要细菌类群。本实验为以后人工湿地系统植物优化选择提供了一定的理论依据。     

Characterization of Microbial Communities and Composition in Constructed Dairy Wetland Wastewater Effluent

Constructed wetlands have been recognized as a removal treatment option for high concentrations of contaminants in agricultural waste before land application. The goal of this study was to characterize microbial composition in two constructed wetlands designed to remove contaminants from dairy washwater. Water samples were collected weekly for 11 months from two wetlands to determine the efficiency of the treatment system in removal of chemical contaminants and total and fecal coliforms. The reduction by the treatment was greatest for biological oxygen demand, suspended solids, chemical oxygen demand, nitrate, and coliforms. There was only moderate removal of total nitrogen and phosphorus. Changes in the total bacterial community and ammonia-oxidizing bacterial composition were examined by using denaturing gradient gel electrophoresis (DGGE) and sequencing of PCR-amplified fragments of the gene carrying the α subunit of the ammonia monooxygenase gene (amoA) recovered from soil samples and DGGE bands. DGGE analysis of wetlands and manure samples revealed that the total bacterial community composition was dominated by bacteria from phylogenetic clusters related to Bacillus, Clostridium, Mycoplasma, Eubacterium, and Proteobacteria originally retrieved from the gastrointestinal tracts of mammals. The population of ammonia-oxidizing bacteria showed a higher percentage of Nitrosospira-like sequences from the wetland samples, while a higher percentage of Nitrosomonas-like sequences from manure, feces, raw washwater, and facultative pond was found. These results show that the wetland system is a natural process dependent upon the development of healthy microbial communities for optimal wastewater treatment.     

Effect of photosynthetic bacteria on water quality and microbiota in grass carp culture

To investigate the effects of photosynthetic bacteria as additives on water quality, microbial community structure and diversity, a photosynthetic purple non-sulfur bacteria, Rhodopseudamonas palustris, was isolated and used to remove nitrogen in the aquaculture water. The results of water quality showed that the levels of ammonia nitrogen, nitrite nitrogen, total inorganic nitrogen and total nitrogen in the treatment group were significantly lower (p < 0.05) than the nitrogen levels of the controls in an extended range. A 454-pyrosequencing analysis revealed that at the level of phylum, Proteobacteria and Firmicutes were dominant in the control group respectively, compared to the dominance of the phyla Proteobacteria, Bacteroidetes and Actinobacteria in the treatment group. The relative abundance of phyla Bacteroidetes and Actinobacteria in treatment witnessed an increase than that in the control. The results also indicated that the treatment group enjoyed a higher microbial diversity than that of the control group. Based on the oxygen requirement and metabolism, the authors observed that the water supplementation with photosynthetic bacteria could significantly decrease (p < 0.05) the number of nitrite reducer and anaerobic bacteria. Therefore, the results suggested that adding photosynthetic bacteria to water improves the water quality as it changes the microbial community structure.     

人工和天然湿地芦苇根际土壤细菌群落结构多样性的比较

为认识人工湿地运行初期根际土壤细菌群落的结构多样性,本研究采用聚合酶链式反应-变性梯度凝胶电泳（PCR-DGGE）的方法,比较了嘉兴石臼漾人工构筑生态湿地和白洋淀天然湿地府河河口区域芦苇根际土壤的细菌群落结构多样性。研究结果显示,嘉兴石臼漾湿地在运行初期（1~1.5年）,根际土壤细菌的物种丰富度已与白洋淀湿地相近,但香农指数和均匀度指数均显著低于白洋淀湿地;随着污染物浓度的降低,土壤细菌群落基因型多样性指数增大。     

Limited Bacterial Diversity within a Treatment Plant Receiving Antibiotic-Containing Waste from Bulk Drug Production

Biological treatment of waste water from bulk drug production, contaminated with high levels of fluoroquinolone antibiotics, can lead to massive enrichment of antibiotic resistant bacteria, resistance genes and associated mobile elements, as previously shown. Such strong selection may be boosted by the use of activated sludge (AS) technology, where microbes that are able to thrive on the chemicals within the wastewater are reintroduced at an earlier stage of the process to further enhance degradation of incoming chemicals. The microbial community structure within such a treatment plant is, however, largely unclear. In this study, Illumina-based 16S rRNA amplicon sequencing was applied to investigate the bacterial communities of different stages from an Indian treatment plant operated by Patancheru Environment Technology Limited (PETL) in Hyderabad, India. The plant receives waste water with high levels of fluoroquinolones and applies AS technology. A total of 1,019,400 sequences from samples of different stages of the treatment process were analyzed. In total 202, 303, 732, 652, 947 and 864 operational taxonomic units (OTUs) were obtained at 3% distance cutoff in the equilibrator, aeration tanks 1 and 2, settling tank, secondary sludge and old sludge samples from PETL, respectively. Proteobacteria was the most dominant phyla in all samples with Gammaproteobacteria and Betaproteobacteria being the dominant classes. Alcaligenaceae and Pseudomonadaceae, bacterial families from PETL previously reported to be highly multidrug resistant, were the dominant families in aeration tank samples. Despite regular addition of human sewage (approximately 20%) to uphold microbial activity, the bacterial diversity within aeration tanks from PETL was considerably lower than corresponding samples from seven, regular municipal waste water treatment plants. The strong selection pressure from antibiotics present may be one important factor in structuring the microbial community in PETL, which may affect not only resistance promotion but also general efficiency of the waste treatment process.     

Comparison of the diversity of root-associated bacteria in Phragmites australis and Typha angustifolia L. in artificial wetlands

Common reed (Phragmites australis) and narrow-leaved cattail (Typha angustifolia L.) are two plant species used widely in artificial wetlands constructed to treat wastewater. In this study, the community structure and diversity of root-associated bacteria of common reed and narrow-leaved cattail growing in the Beijing Cuihu Wetland, China, were investigated using 16S rDNA library and PCR–denaturing gradient gel electrophoresis methods. Root-associated bacterial diversity was higher in common reed than in narrow-leaved cattail. In both plant species, the dominant root-associated bacterial species were Alpha, Beta and Gamma Proteobacteria, including the genera Aeromonas, Hydrogenophaga, Ideonella, Uliginosibacterium and Vogesella. Acidobacteria, Actinobacteria, Nitrospirae and Spirochaetes were only found in the roots of common reed. Comparing the root-associated bacterial communities of reed and cattail in our system, many more species of bacteria related involved in the total nitrogen cycle were observed in reed versus cattail, while species involved in total phosphorus and organic matter removal were mainly found in cattail. Although we cannot determine their nutrient removal capacity separately, differences in the root-associated bacterial communities may be an important factor contributing to the differing water purification effects mediated by T. angustifolia and P. australis wetlands. Thus, further work describing the ecosystem functions of these bacterial species is needed, in order to fully understand how effective common reed- and narrow-leaved cattail-dominated wetlands are for phytoremediation.     

Characterization of microbial communities and composition in constructed dairy wetland wastewater effluent

Constructed wetlands have been recognized as a removal treatment option for high concentrations of contaminants in agricultural waste before land application. The goal of this study was to characterize microbial composition in two constructed wetlands designed to remove contaminants from dairy washwater. Water samples were collected weekly for 11 months from two wetlands to determine the efficiency of the treatment system in removal of chemical contaminants and total and fecal coliforms. The reduction by the treatment was greatest for biological oxygen demand, suspended solids, chemical oxygen demand, nitrate, and coliforms. There was only moderate removal of total nitrogen and phosphorus. Changes in the total bacterial community and ammonia-oxidizing bacterial composition were examined by using denaturing gradient gel electrophoresis (DGGE) and sequencing of PCR-amplified fragments of the gene carrying the alpha subunit of the ammonia monooxygenase gene (amoA) recovered from soil samples and DGGE bands. DGGE analysis of wetlands and manure samples revealed that the total bacterial community composition was dominated by bacteria from phylogenetic clusters related to Bacillus, Clostridium, Mycoplasma, Eubacterium, and Proteobacteria originally retrieved from the gastrointestinal tracts of mammals. The population of ammonia-oxidizing bacteria showed a higher percentage of Nitrosospira-like sequences from the wetland samples, while a higher percentage of Nitrosomonas-like sequences from manure, feces, raw washwater, and facultative pond was found. These results show that the wetland system is a natural process dependent upon the development of healthy microbial communities for optimal wastewater treatment.     

Influence of Season and Plant Species on the Abundance and Diversity of Sulfate Reducing Bacteria and Ammonia Oxidizing Bacteria in Constructed Wetland Microcosms

Constructed wetlands offer an effective means for treatment of wastewater from a variety of sources. An understanding of the microbial ecology controlling nitrogen, carbon and sulfur cycles in constructed wetlands has been identified as the greatest gap for optimizing performance of these promising treatment systems. It is suspected that operational factors such as plant types and hydraulic operation influence the subsurface wetland environment, especially redox, and that the observed variation in effluent quality is due to shifts in the microbial populations and/or their activity. This study investigated the biofilm associated sulfate reducing bacteria and ammonia oxidizing bacteria (using the dsrB and amoA genes, respectively) by examining a variety of surfaces within a model wetland (gravel, thick roots, fine roots, effluent), and the changes in activity (gene abundance) of these functional groups as influenced by plant species and season. Molecular techniques were used including quantitative PCR and denaturing gradient gel electrophoresis (DGGE), both with and without propidium monoazide (PMA) treatment. PMA treatment is a method for excluding from further analysis those cells with compromised membranes. Rigorous statistical analysis showed an interaction between the abundance of these two functional groups with the type of plant and season (p?<?0.05). The richness of the sulfate reducing bacterial community, as indicated by DGGE profiles, increased in planted vs. unplanted microcosms. For ammonia oxidizing bacteria, season had the greatest impact on gene abundance and diversity (higher in summer than in winter). Overall, the primary influence of plant presence is believed to be related to root oxygen loss and its effect on rhizosphere redox.     

辉腾锡勒草原干涸湖泊中氨氧化微生物群落结构分析

【目的】以内蒙古辉腾锡勒草原九十九泉湿地为对象,研究湖泊干涸过程中氨氧化微生物的群落结构及其变化。【方法】通过MPN-PCR定量测定氨氧化古菌(AOA)和氨氧化细菌(AOB)的数量;构建amoA基因克隆文库,进行系统发育分析;结合土壤环境因子,探讨湿地退化过程中影响氨氧化微生物的潜在因素。【结果】依湖泊湿地退水梯度的不同样点中,有75%的样点AOB的数量高于AOA,AOB与AOA的数量比率为0.3-18.1。从湖心到湖岸草原带,AOA和AOB的数量有明显增加,但生物多样性呈降低趋势,二者没有呈现正相关。研究发现,AOB的数量与土壤中NH 4+-N的变化存在良好响应。系统发育分析显示,退化湖泊湿地AOA克隆序列均来自于泉古菌门(Crenarchaeota);AOB的amoA基因的克隆序列大部分与亚硝化单胞菌属(Nitrosomonas)有一定同源性,较少部分与亚硝化螺菌属(Nitrosospira)有一定同源性。【结论】湖泊退水过程增加了湿地土壤氨氧化微生物的数量,而氨氧化微生物的种群丰度有所降低。AOA和AOB群落对湖泊湿地的退化过程做出了响应,其中AOB的响应较为明显,氧化条件和土壤铵浓度的改变可能是促成这种响应的重要原因。     

Effects of plant and influent C:N:P ratio on microbial diversity in pilot-scale constructed wetlands

Microbial processes within the rhizosphere of constructed wetlands are crucial to wastewater treatment, but the relation between microbial community diversity in rhizosphere, plant growth and water quality are unclear at present. The effects of plant growth, water C:N:P ratio and their interaction on microbial diversity in the rhizosphere were studied in synthetic wastewater in planted and unplanted wetlands during three different seasons. The physiological profile of microbial community-level in each wetland was assessed using substrate utilization patterns gathered via BIOLOG? ECO plates. Plant had a significant effect on AWCD parameter, since the planted wetlands usually had a higher the total microbial activity than the unplanted over the study period. The Shannon, Simpson and McIntosh indices in the planted wetlands were apparently higher than those in the unplanted wetlands under any C:N:P ratio influent condition especially in summer. It was also shown that the unplanted wetlands have a greater shift of the interstitial microbial community than the planted at different seasons, since plant rhizospheres produce a more ecologically stable system in order to resist against shifts in microbial community composition in response to C:N:P ratio change in wastewater. Principal component analysis and clustering analysis indicated that influent C:N:P ratio would induce similar microbial species in the planted wetlands and detach them from the unplanted wetlands.