基于16S rRNA基因测序法分析北京霾污染过程中PM2.5和PM10细菌群落特征

2013年1月8~14日,北京出现了严重的霾污染.霾污染时高浓度的大气颗粒物增加了暴露人群的健康风险,而大气中的微生物也可能带来一些风险,但目前对霾污染时大气中微生物组成了解较少.本研究选取了2013年1月8~14日北京7d的PM2.5和PM10采样样本,通过对细菌16S rRNA基因V3区扩增和Mi Seq测序,得到PM2.5和PM10中的细菌群落结构特征,并将结果与相同采样样本的宏基因组测序结果及三项国外基于16S rRNA基因测序方法的大气中细菌研究结果进行了比较.研究发现7 d连续采样条件下,PM2.5中细菌群落结构在门和属的水平上均差别不大.在属级别上,节杆菌属(Arthrobacter)和弗兰克氏菌属(Frankia)是北京冬季大气中细菌群落的主要类群.16S rRNA基因测序与宏基因组测序结果对比分析发现,在属级别上,两种分析方法中有39个相同的属类群(两种分析方法丰度前50的细菌类群合并所得),弗兰克氏菌属(Frankia)和副球菌属(Paracoccus)在16S rRNA基因测序分析结果中相对含量较多,而考克氏菌属(Kocuria)和地嗜皮菌属(Geodermatophilus)在宏基因组测序结果中相对含量较高.在门和属的水平上,PM2.5和PM10中细菌群落结构特征呈现出相似的规律.在门水平上,放线菌门(Actinobacteria)在PM2.5中的相对百分比较大,而厚壁菌门(Firmicutes)在PM10中的相对百分比较大.在属水平上,梭菌属(Clostridium)在PM10中的相对百分比较大.与三项国外基于16S rRNA基因测序研究结果对比发现,尽管在采样地点和采样时间上有较大差异,大气中普遍存在一些相同的细菌类群,且近地面大气细菌群落结构特征相似度较高,区别于高空对流层中细菌群落结构特征.     

典型城市内河菌群组成与氮循环功能垂向分布及溯源分析

微生物在城市河流氮循环中发挥重要作用.由于河流的三维流动性,有必要明晰河流微生物组成和氮循环功能的垂向分布以及水动力因子对微生物来源和群落构建的影响.基于16S rRNA高通量测序技术,研究了北运河北京通州段水体和沉积物的细菌群落组成和氮循环功能,解析了环境要素和水动力因子对群落结构的影响,并对河流细菌进行了溯源.结果表明,沉积物细菌α多样性显著高于水体;从组成上来看,变形菌门(Proteobacteria)丰度最高,在水体和沉积物中的相对丰度分别为54.72%和32.36%. PICRUSt2功能解析表明,北运河有丰富的氮代谢功能,共获得47个氮代谢基因.水体和沉积物表现出相似的功能分布：反硝化作用、氮的同化和异化还原作用相关基因丰度较高,生物固氮和硝化作用相对较低.溯源分析表明,水体细菌来自上游、两岸和沉积物的比例分别为60.05%、37.93%和1.05%,而沉积物细菌来自上游、两岸和上覆水的比例分别为50.16%、45.55%和1.55%,细菌主要通过纵向和横向运移影响当地微生物群落的组成.环境因子、水动力因子和两者相互作用对水体细菌群落变化的解释率分别为44.22%、3.21...     

海水养殖尾水直排对河道沉积物细菌群落的影响

海水养殖活动对周边水域沉积物生态环境的影响日益受到关注,微生物群落是反映环境质量变化的一个重要指标.本研究采集了养殖区进出水河道4个位点(1个进水渠位点、出水口和2个出水渠位点)的沉积物,通过Illumina MiSeq测序技术测定细菌16S rRNA基因,利用磷脂脂肪酸(phospholipid fatty acid,PLFA)技术对沉积物中的微生物群落进行半定量分析,研究海水养殖尾水直接排放对河道沉积物细菌群落结构的影响.结果表明,4个位点中变形菌门(Proteobacteria)为优势细菌类型,主要包括γ-变形菌纲(γ-Proteobacteria)、δ-变形菌纲(δ-Proteobacteria)和α-变形菌纲(α-Proteobacteria).受养殖尾水排放影响,沉积物微生物生物量明显升高,放线菌门相对丰度降低而拟杆菌门相对丰度升高,氨氧化微生物相对丰度降低.非度量多维尺度(non-metric multidimensional scaling,NMDS)分析表明大部分沉积物细菌群落按位点聚类.多样性分析表明,未受养殖尾水影响的沉积物细菌群落多样性高于受养殖尾水影响的位点.冗余分析(redundancy analysis,RDA)表明氨氮与底泥沉积物细菌群落结构的相关性最大,对细菌群落的影响最明显.综上,养殖废水直排明显改变了养殖场河道沉积物物理化学性质和细菌群落结构,说明养殖废水的长期直排已严重污染河道环境,并可能会进一步对近海生态环境造成威胁.     

DNA和cDNA水平对比研究施肥对稻田土壤细菌多样性的影响

依托长沙农业环境观测研究站不同施肥方式长期定位试验,直接从土壤中抽提总DNA和RNA,应用T-RFLP和RTPCR技术在DNA和c DNA水平对比研究施肥对亚热带稻田土壤细菌丰度和群落结构的影响.结果表明,施肥和秸秆还田显著改变了土壤细菌群落组成(DNA水平)和转录组成(c DNA水平)结构,且不同处理中DNA水平的T-RFLP图谱与cDNA水平相差很大.施肥和秸秆还田降低了土壤中存在的细菌多样性,对土壤中转录的细菌多样性没有影响.土壤16S rRNA基因丰度(DNA水平)平均是其转录丰度(c DNA水平)的337倍,与不施氮肥处理(N0)相比,平衡施肥(NPK)没有改变土壤细菌的数量,在平衡施肥的基础上进行秸秆还田增加了土壤细菌的数量,但高量与低量秸秆还田没有显著差异.RDA分析表明,稻田土壤中铵态氮含量是调控各处理土壤中存在及转录的细菌群落的关键因子.总体而言,不管在DNA还是c DNA水平,研究施肥对土壤细菌丰度的影响均是可行并可信的;但只有在c DNA水平开展研究,才能有效发现细菌群落对环境的适应性.     

重金属污染对不同生境中微生物群落结构的影响

重金属污染严重危害了生态系统中微生物群落的多样性和组成结构,然而目前关于重金属污染对“地表水-沉积物-地下水”这3种生境中微生物群落结构的影响差异知之甚少.以唐河污水库地表水、沉积物和地下水为研究对象,基于16S rRNA高通量测序技术分析对比不同生境下微生物群落多样性和组成差异,揭示了3种栖息地中微生物群落结构的主要驱动因素.结果表明,不同生境中微生物群落的多样性具有显著差异,地下水微生物α多样性显著高于沉积物微生物和地表水微生物;不同生境中的微生物物种组成也具有显著差异,地表水中Pedobacter、Hydrogenophaga、Flavobacterium和Algoriphagus丰度较高,沉积物中Ornatilinea、Longilinea、Thermomarinilinea和Bellilinea等重金属耐受菌占据主要优势,而Arthrobacter、Gallionella和Thiothrix是地下水中的标志性物种;理化因素和重金属变量共同驱动了3种生境中微生物的群落结构变化,pH、 NO^-₃-N和Li是影响地表水微生物群落结构的主要...     

基于454和Illumina高通量测序平台对长江口邻近海域沉积物微生物群落的比较分析

作为第二代测序技术的主要平台, 454和Illumina测序方法在微生物核酸分析方面有着广泛应用。本研究以16S rRNA基因为标靶, 分别采用454和Illumina测序技术对长江口邻近海域表层沉积物中的微生物DNA进行分析, 通过对序列长度、微生物群落多样性及组成进行比较, 分析两种方法在沉积物微生物群落结构研究方面的适用性。结果表明, Illumina测序获得的序列数和OTU数要多于454, 其中获得的OTUs中以非优势菌群为主。Illumina与454相比能够检出更多微生物菌群, 且检测到的稀有菌群占有较大比例。综上可知, Illumina测序技术在分析微生物群落多样性及物种组成方面要优于454测序技术, 454的长序列优势并未能体现出来, 且454测序技术可能低估了微生物组成及其多样性。     

典农河沉积物细菌群落结构特征及其与重金属的关系

河流沉积物细菌对有机物和营养盐的转运起着重要作用,其群落结构特征也会受到环境因子的影响.为探究典农河（上游段）沉积物细菌群落结构及其影响因子,于2021年的1、4、7和10月对典农河上游段进行沉积物样品采集调查,测定沉积物中9种重金属（Cu、Zn、Pb、Fe、Mn、As、Cr、Cd和Hg）的含量,基于16S rRNA基因扩增子高通量测序,分析典农河（上游段）不同季节沉积物细菌群落结构特征,通过冗余分析（RDA）识别典农河群落结构变化的驱动因子.结果表明,在空间上,典农河永宁段细菌物种丰度高于典农河银川段细菌物种丰度,在时间上,7月和10月细菌物种丰度高于1月和4月.典农河沉积物细菌最优势菌门为变形菌门（Proteobacteria）,较优势菌门为拟杆菌门（Bacteroidetes）和脱硫菌门（Desulfobacterota）.典农河沉积物细菌优势菌属为Sulfitobacter、 Amylibacter、 Planktomarina、 Sulfurifustis、 SUP05＿cluster、 Arenimonas、Candidatus＿Methanoperedens、Thioal...     

不同种植方式对亚热带红壤微生物多样性的影响

土壤微生物在推动土壤碳循环过程方面发挥着重要作用,然而种植方式的改变对土壤微生物多样性的影响机制还不十分清楚.本研究采集湖南省盘塘县长期定位试验站红壤稻田(PR)、旱地(UC)及水旱轮作(PR)这3种不同种植方式的土壤样品,采用末端限制性酶切片段长度多态性分析(T-RFLP)技术和实时荧光定量(RT-PCR)技术分析了土壤细菌16S rRNA基因的多样性和丰度,研究种植方式改变对土壤微生物数量、群落结构及其多样性的影响.结果表明,3种种植方式的土壤细菌16S rRNA基因数量(以干土计)为2.5×109~1.5×1010拷贝·g-1,与PR相比,UP和UC处理16S rRNA基因丰度显著下降(P0.05).同时,3种种植方式下土壤细菌的优势类群为变形菌(76、90和327 bp;相对丰度47%~53%)和绿弯菌(65 bp;相对丰度10%~12%).冗余分析表明种植方式改变了土壤理化性质,导致土壤细菌群落结构特征的显著变化,而土壤理化性质中有机碳和全氮含量是影响土壤细菌群落结构的主要因子.多样性指数分析(香农指数和均匀度指数)显示种植水稻的土壤细菌多样性最高,显著高于水旱轮作和旱地土壤.可见,种植方式的改变对土壤群落组成和数量造成了深刻的影响,而水稻种植是亚热带红壤可持续利用的一种有效方式,其更有利于土壤有机质的累积,土壤肥力及微生物多样性均较高.     

基于高通量定量PCR与高通量测序技术研究城市湿地公园抗生素抗性基因污染特征

城市湿地公园是城市湿地保护与利用的重要实践,具有重要的生态系统服务价值.抗生素抗性基因(ARGs)作为一种新污染物,在水体环境中普遍存在,是当前环境研究和公众关注热点.基于高通量定量PCR与16S rRNA基因高通量测序技术,深入研究了厦门城市湿地公园水体环境中抗生素抗性基因的种类、丰度水平和分布特征.结合微生物群落特征和水质理化性质,阐释了城市湿地公园抗生素抗性基因污染的影响因子和驱动机制.结果表明,城市湿地公园水体环境总共发现了217种抗生素抗性基因,丰度水平高达6.48×10⁹ copies·L^-1,抗性基因呈现出丰度较高、种类多样以及普遍共存的特点,城市湿地公园是抗生素抗性基因分布的重要热区和存储库;Marivivens、 NS5＿marine＿group和Planktomarina等9种微生物是41种抗生素抗性基因的潜在携带者,微生物群落结构对城市湿地公园水体环境抗性基因变化的单独解释量达到了51%,因此微生物群落是城市湿地抗生素抗性基因赋存和演化最重要的驱动力.基于研究结果,暗示着城市湿地公园水体环境抗生素抗性基因具有潜在水生态安...     

三江平原湿地开垦对土壤微生物群落结构的影响

过度开垦会导致湿地生态系统的快速退化,湿地土壤微生物能够敏感地反映湿地土壤质量及湿地生态系统功能的演变.为研究土地利用变化对湿地微生物群落结构的影响,以黑龙江抚远三江湿地保护区为研究对象,采集其中的原始泥炭湿地、开垦后改种豆科植物及水稻的3种湿地土壤.采用基于细菌16S rRNA基因的高通量测序技术研究上述土壤细菌的群落结构,并探讨其与土壤环境因子间的关系.结果表明,不同土地利用方式湿地土壤中的优势菌门均为变形菌门(Proteobacteria)、放线菌门(Actinobacteria)及酸杆菌门(Acidobacteria),但土地利用方式明显改变了湿地土壤细菌属的组成.改种豆科植物土壤中Blastocatella、Coxiella、Rickettsia丰度较高,水稻田土中Massilia、Nitrosomonas、Bradyrhizobium聚集较多,而泥炭湿地中含较高丰度的Rhizomicrobium、Arthrobacter、Bacillus.结合Chao值与Shannon指数,水稻田土微生物多样性高于改种豆科植物土壤及泥炭湿地土壤,而后两种土壤细菌群落多样性则未见明显差异.相关性分析表明,土壤p H及含水率是影响微生物群落组成的重要驱动因子,说明湿地土壤开垦后改变了土壤p H、含水率及土壤养分,从而对微生物群落结构产生影响.     

Distribution and ecological effect of mercury in Laogang landfill, Shanghai, China

Laogang landfill near Shanghai is the largest landfill in China, and receives about 10000 t of daily garbage per day, Samples of topsoil and plants were analyzed to evaluate mercury pollution from the landfill. For topsoil samples, there were significant correlations among total mercury （HgT）, combinative mercury （Hgc） and gaseous mercury （HgG）, and content of total organic carbon （TOC）, but, no significantly relationship was found between Hg content and filling time. Hg content changes in vertical profiles with time showed that the average Hgv of profiles 1992, 1996, and 2000 was similar, but their average HgG was quite different. HgT was significantly correlated with Hgc in profile 1992 and 2000, and Hgv was significantly correlated with Hg6 in profile 1996. HgG/Hgv ratio in profile samples decreased in the order of （HgG,/HgT）1992〉（HgG/HgT）1996〉〉（HgG/HgT）2000. A simple outline of Hg release in landfill could be drawn： with increasing of filling time, degradation undergoes different biodegradation, accordingly, gaseous mercury goes through small, more, and small proportion to total mercury. Distribution of Hg in plants was inhomogeneous, following the order of leaf〉root〉stem. The highest value of leaf may be associated with higher atmospheric Hg from landfill. Ligneous plants （e.g. Phyllostachys glanca, Prunus salicina and Ligustrum lucidum） are capable of enriching more Hg than herbaceous plants.     

EDTA-enhanced phytoremediation of lead contaminated soil by Bidens maximowicziana

Phytoremediation is a potential cleanup technology for the removal of heavy metals from contaminated soils.Bidens maximowicziana is a new Pb hyperaccumulator,which not only has remarkable tolerance to Pb but also extraordinary accumulation capacity for Pb.The maximum Pb concentration was 1509.3 mg/kg in roots and 2164.7 mg/kg in overground tissues.The Pb distribution order in the B. maximowicziana was:leaf>stem>root.The effect of amendments on phytoremediation was also studied.The mobility of soil Pb and the Pb concentrations in plants were both increased by EDTA application.Compared with CK(control check),EDTA application promoted translocation of Pb to overground parts of the plant.The Pb concentrations in overground parts of plants was increased from 24.23-680.56 mg/kg to 29.07-1905.57 mg/kg.This research demonstrated that B.maximowicziana appeared to be suitable for phytoremediation of Pb contaminated soil,especially,combination with EDTA.     

Decomposition of alachlor by ozonation and its mechanism

Decomposition and corresponding mechanism of alachlor, an endocrine disruptor in water by ozonation were investigated. Results showed that alachlor could not be completely mineralized by ozone alone. Many intermediates and final products were formed during the process, including aromatic compounds, aliphatic carboxylic acids, and inorganic ions. In evoluting these products, some of them with weak polarity were qualitatively identified by GC-MS. The information of inorganic ions suggested that the dechlorination was the first and the fastest step in the ozonation of alachlor.     

Enzymatic oxidation of aqueous pentachlorophenol

The influence of the nonionic surfactant Tween 80 on pentachlorophenol （PCP） oxidation catalyzed by horseradish peroxidase was studied. The surfactant was tested at concentrations below and above its critical micelle concentration （CMC）. Enhancement of PCP removal was observed at sub-CMCs. The presence of Tween 80 in the reaction mixture reduced enzyme inactivation which occurred through a combination of free radical attack and sorption by precipitated products. A simple first-order model was able to simulate time profiles for enzyme inactivation in the presence or absence of Tween 80. At supra-CMCs, the surfactant caused noticeable reductions in PCP removal, presumably through micelle partitioning of PCP which precluded the hydrophobic PCP molecule from interacting with the enzyme.     

基于综合水质标识指数和对应分析法的校园河道水质评价

以三峡大学的校园河道求索溪为研究对象,利用综合水质标识指数法确定求索溪水质类别,分析其水质时空变化规律,并利用对应分析法得出求索溪中不同监测点的主要污染因子.研究结果表明:求索溪整体的综合水质标识指数为7.423,整体水质为劣V类(地表水环境质量标准GB 3838-2002)且黑臭.从时间变化来看,求索溪4月份的水质最差,5月份次之,4、5月份所有监测点的水质都劣于V类且黑臭;8月份水质最好,水质为Ⅳ类;从空间分布来看,8个监测点综合水质标识指数均超过6.0,水质为劣V类,其中6号监测点的水质相对最好,监测点3号的水质相对最差;对应分析法得出求索溪的整体水体污染程度受总氮因子的影响最大,其次为总磷.该研究拟为求索溪及类似校园河道的水环境治理研究提供基础依据和参考.     

Effects of chitosan on growth of an aquatic plant (Hydrilla verticillata) in polluted waters with different chemical oxygen demands

Effects of chitosan on a submersed plant, Hydrilla verticillata, were investigated. Results indicated that H. venicillata could prevent ultrastructure phytotoxicities and oxidativereaction from polluted water with high chemical oxygen demand (COD). Superoxide dismutase (SOD) activity and malondialdehyde (MDA) contents in H. verticillata treated with 0.1% chitosan in wastewater increased with high COD (980 mg/L) and decreased with low COD (63 mg/L), respectively. Ultrastructural analysis showed that the stroma and grana of chloroplast basically remained normal. However, plant cells from the control experiment (untreated with chitosan) were vacuolated and the cell interval increased. The relict of protoplast moved to the center, with cells tending to disjoint. Our findings indicate that wastewater with high COD concentration can cause a substantial damage to submersed plant, nevertheless, chitosan probably could alleviate the membrane lipid peroxidization and ultrastructure phytotoxicities, and protect plant cells from stress of high COD concentration polluted water.     

Removal of cadmium using MnO2 loaded D301 resin

MnO2 loaded weak basic anion exchange resin D301 （Anion exchange resin, macroreticular weak basic styrene） as adsorbent has been prepared and applied to the removal of cadmium. The adsorption characteristics have been investigated with respect to effect of pH, equilibrium isotherms, removal kinetic data, and interference of the coexisting ions. The results indicated that the Cd^2＋ could be efficiently removed using MnO2 loaded D301 resin in the pH range of 3-8 from aqueous solutions with the co-existence of high concentration of alkali and alkaline-earth metals ions. The saturate adsorption capacity of the Cd^2＋ was 77.88 mg/g. The adsorption process followed the pseudo first-order kinetics. The equilibrium data obtained in this study accorded excellently with the Langmuir adsorption isotherm.     

Effect of coagulation pretreatment on the fouling of ultrafiltration membrane

The purpose of this study is to understand the effect and mechanism of preventing membrane fouling, by coagulation pretreatment, in terms of fractional component and molecular weight of natural organic matter （NOM）. A relatively higher molecular weight （MW） of hydrophobic compounds was responsible for a rapid decline in the ultrafiltration flux. Coagulation could effectively remove the hydrophobic organics, resulting in the increase of flux. It was found that a lower MW of neutral hydrophilic compounds, which could remove inadequately by coagulation, was responsible for the slow declining flux. The fluxes in the filtration of coagulated water and supernatant water were compared and the results showed that a lower MW of neutral hydrophilic compounds remained in the supernatant water after coagulation could be rejected by a membrane, resulting in fouling. It was also found that the coagulated flocs could absorb neutral hydrophilic compounds effectively. Therefore, with the coagulated flocs formed on the membrane surface, the flux decline could be improved.     

Penetration analysis of elements and bioleaching treatment of spent refractory for recycling

Acidithiobacillus ferrooxidans ATCC23270 and Acidithiobacillus thiooxidans TM-32 were used for bioleaching of spent refractories of aluminium and copper melting furnaces for their recycling.Firstly,penetration of elements into aluminium melting furnace refractory was investigated and it was found that up to 7 cm from surface was contaminated.Comparison on leaching efficiency by the strains ATCC23270 and TM-32 found that the strain ATCC23270 could treat larger amount of the refractories than the strain TM-32 could do.In the experiment of bioleaching of spent refractory aluminium melting furnace by the strain ATCC23270,high leaching efficiency were obtained on Al,Si,and Ca,and extremely low leaching performance was,however,shown on the rest of elements i.e.,Na,Mn,and Zn.Under the strain TM-32 use,relatively high leaching performance was recognized on Al,Si,Ca,Na,Mn,and Zn.In the experiment of bioleaching for spent refractory copper melting furnace,almost the same leaching trends were shown on Cu,Zn,Al,and Si under the strains ACTT23270 and TM-32 uses.     

Simultaneous removal of ammonium and phosphate by zeolite synthesized from coal fly ash as influenced by acid treatment

Zeolite synthesized from fly ash （ZFA） without modification is not efficient for the purification of NH4＋ and phosphate at low concentrations that occur in real effluents, despite the high potential removal capacity. To develop an effective technique to enhance the removal efficiency of ammonium and phosphate at low concentrations, ZFA was modified with acid treatment and the simultaneous removal of ammonium and phosphate in a wide range of concentration was investigated. It was seen that when compared with untreated ZFA, only the treatment by 0.01 mol/L of H2SO4 significantly improved the removal efficiency of ammonium at low initial concentrations. The behavior was well explained by the pH effect. Treatment by more concentrated H2SO4 led to the deterioration of the ZFA structure and a decrease in the cation exchange capacity. Treatment by 0.01 mol/L H2SO4 improved the removal efficiency of phosphate by ZFA at all initial P concentrations, while the treatment by concentrated H2SO4 （≥0.9 mol/L） resulted in a limited maximum phosphate immobilization capacity （PIC）. It was concluded that through a previous mild acid treatment （e.g. 0.01 mol/L of H2SO4）, ZFA can be used in the simultaneous removal of NH4＋ and P at low concentrations simulating real effluent.