自然微生物挂膜处理水产养殖废水的效果及微生物群落分析

以弹性填料和流化床填料为硝化反应的生物挂膜材料, 聚羟基丁酸/戊酸共聚酯(PHBV)为反硝化反应的碳源和生物膜载体, 通过微生物自然挂膜处理低C/N比水产养殖废水, 去除水体中的氨氮、亚硝酸盐氮及总氮。应用Miseq高通量测序技术对生物膜的微生物群落组成和结构进行分析。结果表明: 温度25—30℃, 该处理系统首次挂膜成功需要4周, 启动后运行稳定, 对2种不同来源和氮污染程度的养殖废水均有较好的脱氮效果, 氨氮、亚硝酸盐氮及总氮的去除率均在90%以上。硝化生物膜(a)的优势菌分别归属变形菌门(Proteobacteria)、拟杆菌门(Bacteroidetes)和厚壁菌门(Firmicutes)。反硝化生物膜(b)微生物群落的多样性指数和丰度指数均远大于前者, 主要为变形菌门、厚壁菌门、拟杆菌门、螺旋体门(Spirochaetae)及绿菌门(Chlorobi)。其中, 归属于变形菌门β-变形菌纲(Betaproteobacteria)的丛毛单胞菌科(Comamonadaceae)和红环菌科(Rhodocyclaceae)在2种生物膜中占比均较高。由于所处环境(载体, 碳源、溶氧等)不同, 在属分类水平上, 2种生物膜的细菌群落结构表现出明显差异。生物膜a中属的种类仅为b的三分之二, 相对丰度>0.5%的优势菌属, a为8个, b为18个。其中, 隶属丛毛单胞菌科和红环菌科未知属的优势种群分别占到a、b总序列数的56.67%和45.51%。磁螺菌属(Magnetospirillum)和硝化螺菌属(Nitrospira)是a中特有的优势功能菌群, 梭菌属(Clostridium)、动胶菌属(Zoogloea)、管道杆菌属(Cloacibacterium)、脱硫弧菌属(Desulfovibrio)等具有反硝化功能的菌群为b的优势菌属。     

好氧-厌氧混合污泥启动微生物燃料电池产电性能及微生物群落动态特征

【目的】为探讨好氧-厌氧混合污泥启动微生物燃料电池(Microbial fuel cell,MFC)产电性能以及MFC对微生物群落的选择作用,【方法】以乳酸为底物,应用不依赖于培养的微生物分子生物学技术解析单室MFC启动过程中微生物群落的组成和结构动态学特征。【结果】结果表明,MFC经过3个周期启动成功,最高输出电压230 m V。当MFC外电阻为1656Ω时,最大功率密度11.15 W/m3,电池运行稳定。混合污泥启动MFC以后,阳极生物膜微生物群落结构同种泥差异较大,且多样性降低。生物膜中微生物类群按丰度依次为β-变形菌纲(Betaproteobacteria)24.90%、拟杆菌门(Bacteroidetes)21.30%、厚壁菌门(Firmicutes)9.70%、γ-变形菌纲(Gammaproteobacteria)8.50%、δ-变形菌纲(Deltaproteobacteria)7.90%、绿弯菌门(Chloroflexi)4.20%以及α-变形菌纲(Alphaproteobacteria)3.60%。有利于生物膜形成与稳定的动胶菌属(Zoogloea)和不动杆菌属(Acinetobacter)序列丰度分别占生物膜群落的5.00%和3.90%,与MFC产电能力直接相关的地杆菌属(Geobacter)序列由混合污泥中的0.60%上升至阳极生物膜中的2.60%。【结论】本研究表明,MFC阳极生物膜在驯化过程中对污泥中的微生物进行淘汰和选择,最终驯化形成了有利于生物膜形成与稳定、有机物厌氧发酵与产电的微生物菌群。     

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。     

生物膜法强化净化氨氮污染水体及其微生物群落解析

【目的】比较不同营养条件及挂膜方式下生物膜法对氨氮污染水体的净化效果及其功能微生物群落结构。【方法】设置空白(Blank)、自然成膜(Raw)、预附脱氮菌强化挂膜(PCC)3组生物膜反应器,利用末端限制性片段长度多态性(T-RFLP)技术和非度量多维标度(NMDS)分析方法对生物膜反应器转化氨氮过程中微生物群落结构及其演替过程进行动态解析。【结果】在C/N=1:1时,除PCC在起始阶段短暂具有较高的氨氮脱除效率外,Blank、Raw和PCC最终均表现出较低的氨氮转化效率(10%-20%)。改变C/N=2:1后,Raw和PCC对人工合成污水中NH4+-N的转化率均提高至95%以上,而且Raw与PCC的群落结构在C/N=2:1时具有较高的相似性,优势菌群主要为γ-变形菌纲(Gammaproteobacteria)、放线菌纲(Actinobacteria)和硝化螺菌纲(Nitrospira)。【结论】C/N是影响生物膜反应器氨氮去除效果及驱动生物膜反应器中细菌群落结构发生改变的重要因子。     

退耕还湿后土壤细菌群落结构和生物量变化过程

土壤细菌对湿地生态系统功能和健康维持起着重要作用。以菜子湖原始湿地、不同退耕年限湿地(3a、7a、11a和21a)和仍耕作油菜地土壤为研究对象,应用聚合酶链式反应-变性梯度凝胶电泳(PCR-DGGE)、高通量测序和磷脂脂肪酸(PLFAs)方法分析土壤细菌群落组成和生物量,探讨它们在退耕还湿后的变化过程及其影响因素。结果表明,退耕还湿后土壤变形菌门(α-变形菌纲、β-变形菌纲和δ-变形菌纲)、酸杆菌门(酸杆菌纲和全噬菌纲)、Nitrospinae(Nitrospinia纲)和硝化螺旋菌门(硝化螺旋菌纲)相对丰度先增高后降低;这些参与氮循环的土壤细菌对退耕后湿地生态恢复过程中土壤氮素提升起着重要作用。与农业生产活动密切关联的厚壁菌门(芽孢杆菌纲和梭菌纲)和放线菌门(放线菌纲)相对丰度逐渐降低。湿地土壤细菌多样性在退耕初期(3—7a)上升达到最大,退耕中后期逐渐降低。表层土壤各类群细菌生物量逐渐升高,亚表层土壤各类群细菌生物量则先降后升再降。水分条件和容重是与研究区土壤细菌群落结构和多样性密切相关的土壤因子,而全氮是与土壤细菌生物量密切相关的土壤因子。研究从生态过程视角解析了土壤细菌群落较详...     

京津冀区域市政污水厂活性污泥种群结构的多样性及差异

【背景】活性污泥中微生物的种群结构影响着污水生物处理的高效性及稳定性,是有效保证污水处理效果的关键。【目的】研究活性污泥中细菌的群落结构组成及多样性,并分析相应菌群的主要功能,旨在更好地发挥细菌的净化作用、保持污水处理过程的稳定及提高污水的处理效率。【方法】以京津冀区域内典型市政污水厂活性污泥为研究对象,通过IlluminaMiSeq高通量测序及实时定量PCR技术,对5个污水厂活性污泥的微生物种群结构特征进行了详细解析,研究不同工艺参数下活性污泥中优势种群及脱氮菌群丰度的差异。【结果】5个污水厂活性污泥种群结构具有一定差异,其中Hengshui (HS)厂污泥的群落结构受温度的影响最大,而Shahe (SH)、Daoxianghu (DXH)、Nangong(NG)厂活性污泥群落结构则受总氮、总磷与氨氮的共同影响,氨氮对SH厂活性污泥种群结构影响最大。DXH、NG和HS厂污泥中优势菌均为Anaerolineaceae,而SH和Hejian (HJ)厂的优势菌则为Saprospiraceae与Lactobacillus。活性污泥中反硝化菌丰度最高的为HJ厂,丰度最低的为HS厂,反硝化功能基因nirS比nirK分布更为广泛。【结论】对于不同污水厂,影响其活性污泥群落结构组成的环境因素也是不同的,并且特殊的进水水质也会对污泥菌群组成和生物多样性产生影响。     

以聚羟基丁酸戊酸共聚酯为碳源去除循环水养殖系统的硝酸盐及生物膜中微生物群落动态

【目的】利用聚羟基丁酸戊酸共聚酯(PHBV)作为固体碳源和生物膜载体,去除循环水养殖系统的硝酸盐,并研究固体碳源反硝化反应器不同运行阶段生物膜中微生物群落结构的动态变化。【方法】采用PCR-DGGE技术对反硝化反应器中微生物群落结构的动态变化进行了分析,采用传统纯培养方法分离反应器中降解PHBV的细菌。【结果】连接固相反硝化反应器能使循环水养殖系统中积累的硝酸盐显著降低,并维持在较低水平(小于10 mg/L),而常规循环水养殖系统中硝酸盐浓度持续增加。系统发育树聚类分析结果表明,反硝化反应器生物膜细菌归属于变形菌门(β-proteobacteria、γ-proteobacteria和δ-proteobacteria)、厚壁菌门(Firmicutes)和拟杆菌门(Bacteroidetes)。反应器运行初期(40 d)的优势种群主要为Acidovorax和Bacillus;运行后期(150 d)的优势种群依次为Clostridium、Desulfitobacterium、Dechloromonas、Pseudoxanthomonas和Flavobacterium。从反应器中分离到的PHBV降解菌株分别归属于Acidovorax、Methylibium、Pseudoxanthomonas和Dechloromonas。【结论】利用PHBV为碳源能有效去除循环水养殖系统的硝酸盐。明确了反硝化反应器在运行过程中,碳源表面生物膜的优势菌群及其动态变化。     

基于高通量测序的4种硝化细菌富集培养物微生物群落结构分析

【目的】比较分析4种硝化细菌富集培养物(以铵盐为氮源的淡水富集物A、以亚硝酸盐为氮源的淡水富集物B、以铵盐为氮源的低温淡水富集物C和以亚硝酸盐为氮源的海水富集物D)的微生物群落结构与组成。【方法】分别提取样品的总DNA,采用高通量测序技术,分析微生物群落的组成、丰度和多样性。【结果】在不同微生物的分类水平,4个样品共检测到24门47纲129属。4个样品的优势菌门均为变形菌门;样品A、B、C的优势菌纲为β-变形菌纲和γ-变形菌纲,样品D的优势菌纲为γ-变形菌纲、δ-变形菌纲和芽孢杆菌纲;而优势菌属各不相同,其中样品A为亚硝化单胞菌属(24.56%),样品B为链霉菌属(7.15%),样品C为噬菌弧菌属(19.36%)和类诺卡氏菌属(19.35%),样品D为嗜酸菌属(13.6%)和柄杆菌属(11.5%)。共检测出7种具有硝化功能的细菌,其中样品A、B和D中主要是亚硝化单胞菌属,占比分别为24.56%、4.94%和0.63%,样品C主要为Nitrospirillum(0.69%)和硝化螺旋菌属(0.69%)。此外在样品中还检测到红灯食烷菌、羽扇豆根瘤菌等有益菌,以及弧菌属、伯克霍尔德菌等致病菌。【结论】阐述了4个样品微生物群落结构的多样性,确定了不同培养物中起主要作用的硝化细菌类群以及其它与环境物质循环相关或具有特殊生理特性的菌群,研究结果为硝化细菌富集培养物的实际应用奠定了基础。     

甘肃小陇山油松与柴胡栽培土壤细菌群落特征

土壤细菌能够有效促进土壤中物质循环和能量流动,其群落组成、数量及多样性能够反映土壤质量状况。研究不同植被对土壤细菌群落结构和多样性的影响,有助于深入了解土壤健康状况,对实现植被管理和土地可持续利用具有重要意义。以甘肃小陇山移栽至耕地(耕地前期种植柴胡)后生长4年的人工油松苗(Pinus tabulaeformis Carr)土地和长期种植柴胡(Bupleurum chinense)土地的表层土壤(0—30 cm)为对象,采用Illumina Miseq高通量测序技术对2种人工植被下土壤细菌群落进行测定,分析了土壤细菌的Alpha多样性、群落组成和丰富度,并研究土壤化学性质对细菌群落结构的影响。结果表明:1)两种不同植被的土壤细菌群落丰度和多样性无明显差异,但油松地土壤细菌群落均匀度明显小于柴胡地土壤。2)所测土壤样品中共检测到细菌的37个门,84个纲,168个目;其中,主要的优势菌门为变形菌门(Proteobacteria),酸杆菌门(Acidobacteria),绿弯菌门(Chloroflexi),浮霉菌门(Planctomycetes),放线菌门(Actinobacteria),且油松地土壤中的变形菌门(Proteobacteria)的丰富度显著低于柴胡地土壤(P 0. 05),绿弯菌门(Chloroflexi)丰富度却显著高于柴胡地(P 0. 05);主要的优势菌纲为酸杆菌纲(Acidobacteria),β-变形菌纲(Betaproteobacteria),放线菌纲(Actinobacteria),变形菌纲(Alphaproteobacteria)和芽单胞菌纲(Gemmatimonadetes),但各优势菌纲的丰度在两个植被土壤之间差异不显著(P0.05)。3)变形菌门(Proteobacteria)丰富度与土壤的碱解氮含量呈显著负相关(P0.05),酸杆菌门(Acidobacteria)丰富度与全氮含量显著负相关(P0.01);通过RDA分析发现,碱解氮和全氮是影响细菌群落结构的最主要因子。可见,油松和柴胡植被土壤细菌群落结构及多样性差异不大,但油松地土壤细菌群落的物种分配较柴胡地集中,且优势细菌门中的变形菌门和绿弯菌门在两者之间的差异显著,同时,碱解氮和全氮含量是影响2种植被土壤细菌群落的主要因子。这些结论为了解小陇山不同植被土壤微生物群落组成及多样性提供理论,也为当地进行人工油松苗的种植、经营及土壤管理提供参考。     

泰山4种优势造林树种细根分解对细菌群落结构的影响

为了理解细菌群落结构和多样性对森林生态系统细根凋落物分解的影响, 该研究以泰山4种主要优势造林树种刺槐(Robinia pseudoacacia)、麻栎(Quercus acutissima)、油松(Pinus tabulaeformis)和赤松(Pinus densiflora)为研究对象, 采用凋落物分解袋法及Illumina Miseq测序平台对细菌16S rDNA V4-V5区扩增产物进行双端测序, 分析了4种树种细根分解对细菌群落结构及多样性的影响。结果表明: (1) 4种植物细根分解速率差异显著, 阔叶树种分解速率显著高于针叶树种, 表现为刺槐>麻栎>油松>赤松。(2) 4个树种细菌序列操作分类单元(OTU)、观测到的物种数、Ace指数和系统发育多样性之间差异显著, 且阔叶树种刺槐和麻栎显著低于针叶树种赤松和油松。4种细根分解的细菌群落结构存在极显著差异。细根初始碳(C)含量、木质素:氮(N)和C:N对细菌群落结构的影响较大。(3)细菌群落相对丰度在5%以上的优势类群是变形菌门、放线菌门、拟杆菌门、酸杆菌门, 且变形菌门、酸杆菌门在4个树种之间差异显著, 特别是阔叶树种变形菌门显著高于针叶树种。在纲水平上, α-变形菌纲、β-变形菌纲、γ-变形菌纲、不明放线菌纲、鞘脂杆菌纲为主要的优势纲, 其中α-变形菌纲、不明放线菌纲在4个树种之间差异显著。(4) Pearson相关性分析表明, 细菌优势门和纲相对丰度受到凋落物初始化学性质的影响, 特别是变形菌门和α-变形菌纲; 变形菌门和α-变形菌纲相对丰度与细根分解速率显著正相关。冗余分析结果也显示, 细根初始N、磷(P)含量和木质素含量对细菌群落结构的影响较大。研究结果有助于理解细菌群落结构和多样性对森林生态系统细根凋落物分解的影响。     

A Unique Protease Cleavage Site Predicted in the Spike Protein of the Novel Pneumonia Coronavirus (2019-nCoV) Potentially Related to Viral Transmissibility

正Dear Editor,In December 2019, a novel human coronavirus caused an epidemic of severe pneumonia(Coronavirus Disease 2019,COVID-19) in Wuhan, Hubei, China(Wu et al. 2020; Zhu et al. 2020). So far, this virus has spread to all areas of China and even to other countries. The epidemic has caused 67,102 confirmed infections with 1526 fatal cases     

Curcuminoids as inhibitors of thioredoxin reductase: A receptor based pharmacophore study with distance mapping of the active site

Curcumin is the yellow pigment of turmeric that interacts irreversibly forming an adduct with thioredoxin reductase (TrxR), an enzyme responsible for redox control of cell and defence against oxidative stress. Docking at both the active sites of TrxR was performed to compare the potency of three naturally occurring curcuminoids, namely curcumin, demethoxy curcumin and bis-demethoxy curcumin. Results show that active sites of TrxR occur at the junction of E and F chains. Volume and area of both cavities is predicted. It has been concluded by distance mapping of the most active conformations that Se atom of catalytic residue SeCYS498, is at a distance of 3.56 from C13 of demethoxy curcumin at the E chain active site, whereas C13 carbon atom forms adduct with Se atom of SeCys 498. We report that at least one methoxy group in curcuminoids is necessary for interation with catalytic residues of thioredoxin. Pharmacophore of both active sites of the TrxR receptor for curcumin and demethoxy curcumin molecules has been drawn and proposed for design and synthesis of most probable potent antiproliferative synthetic drugs.     

Comparative morphology of the carpel in the Liliaceae: Hewardieae, Petrosavieae, and Tricyrteae

The young pistils in the melanthioid tribes, Hewardieae, Petrosavieae and Tricyrteae, are uniformly tricarpellate and syncarpous. They lack raphide idioblasts. All are multiovulate, with bitegmic ovules. The Petrosavieae are marked by the presence of septal glands and incomplete syncarpy. Tepals and stamens adhere to the ovary in the Hewardieae and the Petrosavieae but not in the Tricyrteae. Two vascular bundles occur in the stamens of the Hewartlieae and Tricyrtis latifolia. Ventral bundles in the upper part of the ovary of the Hewardieae are continuous with compound septal bundles and placental bundles in the lower part. Putative ventral bundles occur in the alternate position in the Tricyrteae and putative placental bundles in the opposite. position in the Petrosavieae. The dichtomously branched stigma in each carpel of the Tricyrteae is supplied by a bifurcated dorsal bundle.     

Enterovirus 71 3C proteolytically processes the histone H3 N-terminal tail during infection

Highlights
1. The N-terminal tail of histone H3 is specifically cleaved during EV71 infection.
2. Viral protease 3C is identified as a protease responsible for proteolytically processing the N-terminal H3 tail.
3. Our finding reveals a new epigenetic regulatory mechanism for Enterovirus 71 in virus-host interactions.     

Detection of EBV and HHV6 in the Brain Tissue of Patients with Rasmussen’s Encephalitis

Rasmussen’s encephalitis (RE) is a rare pediatric neurological disorder, and the exact etiology is not clear. Viral infection may be involved in the pathogenesis of RE, but conflicting results have reported. In this study, we evaluated the expression of both Epstein-Barr virus (EBV) and human herpes virus (HHV) 6 antigens in brain sections from 30 patients with RE and 16 control individuals by immunohistochemistry. In the RE group, EBV and HHV6 antigens were detected in 56.7% (17/30) and 50% (15/30) of individuals, respectively. In contrast, no detectable EBV and HHV6 antigen expression was found in brain tissues of the control group. The co-expression of EBV and HHV6 was detected in 20.0% (6/30) of individuals. In particular, a 4-year-old boy had a typical clinical course, including a medical history of viral encephalitis, intractable epilepsy, and hemispheric atrophy. The co-expression of EBV and HHV6 was detected in neurons and astrocytes in the brain tissue, accompanied by a high frequency of CD8⁺ T cells. Our results suggest that EBV and HHV6 infection and the activation of CD8⁺ T cells are involved in the pathogenesis of RE.     

Perinatal Vertical Transmission of Chikungunya Virus in Ruili,a Town on the Border between China and Myanmar

正Dear Editor,Chikungunya virus (CHIKV), an arbovirus in the family of Togaviridae, genus Alphavirus, is transmitted by the A.aegyptii or A. albopictus mosquito, and causes disease in humans characterized by fever, rash, and arthralgia (Silva and Dermody 2017; Suhrbier 2019). It was first reported in 1953 in Tanzania, and caused only a few outbreaks and sporadic cases in Africa and Asia in last century. However, in the epidemic in 2004, CHIKV acquired mutations that conferred enhanced transmission by the A. albopictus mosquito(Schuffenecker et al. 2006). Since then, it has successively caused outbreaks in Africa, the Indian Ocean, South East Asia, the South America, and Europe (Zeller et al. 2016).     

Development of a Novel Reverse Transcription Loop-Mediated Isothermal Amplification Method for Rapid Detection of SARS-CoV-2

In conclusion, the novel visual RT-LAMP assay is a simple, rapid, and sensitive approach for detection of SARS-CoV-2, and it is ready for application in primary care and community hospitals or health care centers, and even patients' own houses in response to the current SARS-CoV-2 epidemic because the assay does not require sophisticated equipment and skilled personnel. Furthermore, it is also ready to be used in fields for screening samples from wild animals and environments to facilitate the identification of potential intermediate hosts that mediate the cross-species transmission of SARS-CoV-2 from bats to humans.