太湖蓝藻水样中藻蓝蛋白提取方法比较

以2011年8月20日采集的太湖梅梁湾的夏季蓝藻水华为研究对象,通过12个样点平行样的藻蓝蛋白实验提取,基于光谱吸收特征和浓度值,对反复冻融法、超声波法、溶胀法、丙酮法的提取效果进行比较评价.结果表明:4种方法获取的藻蓝蛋白提取液在620 nm附近出现吸收峰,其中,反复冻融法的峰高最强,超声波法最弱,说明反复冻融法的提取效果优于其他方法;反复冻融法、超声波法、溶胀法获取的部分蓝蛋白提取液在670 nm附近具有次吸收峰,与藻蓝蛋白标样的吸收曲线存在差异;反复冻融法、超声波法提取的藻蓝蛋白浓度值变异系数小于0.6,表明这两种方法较其他方法稳定;反复冻融法提取的藻蓝蛋白浓度值高于其他3种方法,可推荐作为太湖蓝藻水样中藻蓝蛋白的提取方法.     

巢湖蓝藻水华藻胆蛋白的分离与鉴定

在高温季节,巢湖等国内淡水湖中蓝藻大量繁殖常形成蓝藻湖靛(主要由微囊藻组成),它含有丰富的蛋白质、氨基酸、脂肪等营养成分.本文对微囊藻中氨基酸的组成及含量进行了测试,并从其中提取、分离纯化了藻胆蛋白(指其中的藻蓝蛋白和别藻蓝蛋白),最后运用紫外可见光谱对其进行了检测,取得了较为满意的结果.这也为进一步开发利用巢湖湖靛提供了依据。     

基于矩阵模型进行抗荧光干扰的蓝藻原位检测方法

藻蓝蛋白与叶绿素a作为特征色素常用于表征蓝藻的生物量.藻蓝蛋白与叶绿素a通常用荧光光谱或原位荧光强度检测,但是在实际水体中藻蓝蛋白与叶绿素a的荧光效应相互干扰会影响测量精度,极大限制了该方法的应用.本文根据朗伯比尔定律,利用二阶矩阵模型设计了一套在荧光检测过程中"激发光-发射光"优化抗干扰的波长选择方法,并利用多元线性回归分析建立了两种色素浓度与荧光强度之间的多元校正线性模型,实验验证了所筛选蓝藻荧光分析法中特定激发光和发射光波长的有效性.本研究实现了两种色素检测过程中的成功解耦,可在藻蓝蛋白的原位检测中排除水体中藻蓝蛋白与叶绿素a之间的相互干扰,提高藻蓝蛋白和叶绿素a的原位检测精度,为开发便携式蓝藻检测传感器时的波长选择提供理论基础.     

富营养化湖泊典型水华蓝藻的固有光学特性

固有光学特性是水体光学特性的重要内容,掌握富营养化湖泊水体内典型水华蓝藻的固有光学特性,是开展不同水华蓝藻遥感识别的理论基础.利用AC-S吸收衰减仪、BB9后向散射仪,通过实验室纯藻培养,研究微囊藻（Microcystis）、鱼腥藻（Dolichospermum）和束丝藻（Aphanizomenon）3种典型水华蓝藻的固有光学特性,并探讨色素浓度、色素占比以及藻类等效粒径对不同水华蓝藻固有光学特性的影响.结果表明,3种典型水华蓝藻的吸收光谱曲线均具有440、620和675 nm吸收峰,微囊藻620和675 nm的比吸收系数最大,鱼腥藻440 nm处的比吸收系数最大;束丝藻单位色素浓度的散射和后向散射能力最高,鱼腥藻次之,微囊藻最低;固有光学特性影响因子分析表明,色素浓度和藻蓝素占比是影响3种水华蓝藻固有光学特性的主要因素.3种蓝藻的吸收系数、散射系数以及鱼腥藻、束丝藻的后向散射系数均随着色素浓度（叶绿素a或藻蓝素）的增加而增大;当蓝藻中藻蓝素占比增加时,3种蓝藻的单位色素浓度的后向散射系数逐渐下降;而藻细胞粒径与固有光学特性之间并未表现出很好的相关性.因此,3种水华蓝藻单位色素浓度的固有光学特性将为典型水华蓝藻的遥感识别提供重要的理论基础和数据支持.     

一株来源于水库的浮游蓝丝藻SK3株的生长及产毒特性

浮游蓝丝藻是水华蓝藻的常见种属,但对其生长和产毒特性认识不多本文首次对我国水体中的浮游蓝丝藻进行了研究.从广东省一重要供水水库中分离到一株丝状蓝藻,经形态特征鉴定为阿氏浮游蓝丝藻,并在不同的光照和温度下对该藻株进行了培养.结果表明,适合于该藻生长的光强和温度分别为20μmol/(m~2·s)和30℃;该藻株具有mcyE基因,能产生微囊藻毒素MCRR,产量为0.06μg/mg(DW).     

基于Sentinel-3 OLCI影像的巢湖藻蓝素浓度年内动态遥感监测

藻蓝素(PC)是水体蓝藻的指示性色素,其浓度反映了蓝藻生物量,利用卫星遥感监测藻蓝素浓度年内动态对蓝藻水华的有效防控有着重要意义.根据不同季节的巢湖藻蓝素浓度实测数据与同期Sentinel-3 OLCI影像,构建机器学习回归反演模型,应用于巢湖2019年OLCI影像集上,对巢湖藻蓝素浓度的空间分布、年内变化进行遥感监测...     

基于水华蓝藻固有光学特性的主要类群定量识别方法

蓝藻水华是湖泊水体富营养化的重要特征之一,不同水华蓝藻类群形成的水华特征、危害及其治理方法差异显著.因此,如何快速、准确地掌握不同蓝藻类群的时空分布特征成为实施富营养化湖泊污染治理与生态恢复、蓝藻生态灾害预测预警中一个亟待解决的科学问题.本研究基于纯藻种实验室培养和室内光学控制实验,在微囊藻(Microcystis)、鱼腥藻(Dolichospermum)、束丝藻(Aphanizomenon)3种主要水华蓝藻固有光学特性的基础上,通过甄别不同水华蓝藻的吸收、散射和后向散射光谱的特征波段,构建了基于吸收和散射特性的5种水华蓝藻类群的非线性最优化定量识别模型,其中,基于440、620和675 nm 3个波段吸收的a-CIM 440,620,675具有较为稳定的定量识别能力;并基于野外实测光学特性数据,实现了巢湖主要水华蓝藻类群的定量监测,初步分析了巢湖主要水华蓝藻类群的时空分布特性.研究表明,巢湖的水华蓝藻以鱼腥藻、微囊藻为主,束丝藻较少,鱼腥藻主要出现在温度较低的季节,微囊藻在夏季的西部湖区占优势;巢湖水华主要为微囊藻藻华和鱼腥藻藻华,且浓度较高的蓝藻主要存在于水体表面以下20 cm范围内;微囊藻和鱼腥藻在非藻华断面垂向上均匀分布.本研究可为富营养化湖泊蓝藻水华预测预警以及相关管理部门决策提供重要的理论依据和科学支撑.     

蓝藻不同类型胶被提取方法

胶被作为蓝藻细胞外的一层功能性结构,其提取方法仍不清晰完善.本研究选取具有不同胶被形态的鱼腥藻Anabaena sp.和念珠藻Nostoc sp.为实验材料,通过归纳近10年有关藻类胶被的提取方法,总结影响胶被提取效果的主要因素(温度、提取剂浓度、时间等)范围,并设置一系列因素及其梯度的双因素实验和正交实验对胶被提取方法进行优化,观察提取过程中藻细胞形态、自发荧光以及藻细胞密度变化.结果表明,粘液可通过离心法与藻细胞分离;荚膜提取的硫酸水浴法的最优条件为:1 mol/L、60℃、30 min,氢氧化钠水浴法为:0.01 mol/L、50℃、20 min;鞘提取的最优方法条件为氢氧化钠水浴法(0.01 mol/L、80℃、40 min)和超声法(20 kHz,30 W,on:3 s,off:3 s,2 min)结合处理.此外,通过高效液相色谱法测定提取的蓝藻胶被多糖组分,结果显示,鞘具有较强的疏水性,未水化为粘液或RPS,而荚膜中含有的氨基葡萄糖(1.261±0.02 mg/g)使得荚膜比鞘更具有粘附力.将本研究得到的荚膜和鞘的最优提取法运用到胶被形态以荚膜和鞘为主的其他藻种,均有很好的提取效果.本研究可为后续进一步揭示不同类型胶被的功能作用提供技术支撑.     

秋季聚积蓝藻打捞对蓝藻生长及水质影响的原位实验

在巢湖西北半湖近岸设置3组小型围隔模拟秋季湖岸带蓝藻聚积,并用单片鳃式过滤器原位打捞蓝藻,研究秋季打捞对蓝藻生长的影响及其对营养盐、藻源性有机物的控制效应.初始围隔水体叶绿素a浓度为309.5±3.7μg/L,总氮和总磷浓度分别为3.32±0.14和0.30±0.04 mg/L.蓝藻衰亡分解释放的藻源性有机物为水体溶解性有机物的主要来源,荧光有机物以类蛋白物质为主.经过打捞,浮游植物生物量削减了41.7%,解除了蓝藻生长"密度制约",24 h细胞分裂频率及原位生长速率均增大,说明打捞在短期内增强蓝藻细胞生长活力,减缓藻源性有机物的释放.与秋季蓝藻衰亡趋势一致,实验周期内围隔中叶绿素a浓度逐渐降低,秋季打捞不会造成蓝藻水华再次暴发.打捞通过削减蓝藻生物量,使水体初级生产力、氮、磷、高锰酸盐指数得到显著控制;而且打捞还可以控制藻源性有机物的释放,使藻源性大分子有机物更易降解为小分子有机物.因此,在秋季对湖岸带聚积蓝藻进行物理打捞,不仅可以控制蓝藻生物量,还可以有效控制营养盐和有机物的释放,降低生态风险.     

基于生态-经济重要性的滨湖城市土地开发适宜性分区研究——以无锡市为例

契爷石水库是广东省东莞市的一座中型供水水库,为富营养化水体.为改善该水库水质,2002年10月开始从东江调水,以满足供水的水量和水质要求.在调水后的两年期间,对水文、水质和浮游植物种类和数量进行了调查.本文将调查结果与以往(2000年)调查数据进行了对比,分析了调水后该水库水质和浮游植物的特点和调水对水质的影响.调水直接导致TP浓度显著下降,丰水期的TN浓度有一定下降,但调水并没显著降低枯水期TN的浓度.调水后,丰水期水体的叶绿素a浓度有明显的增加,枯水期叶绿素a浓度大大低于调水前.与调水前相比,调水后浮游植物种类数有明显增加,特别是绿藻种类,蓝藻中的假鱼腥藻和针状蓝纤维藻仍为浮游植物优势种,但硅藻中的短小曲壳藻和针杆藻等成为新的优势种,裸藻的数量大为减少.     

Performance of grass and eucalyptus riparian buffers in a pasture catchment,Western Australia,part 2: water quality

Declining water quality on the south coast of Western Australia has been linked to current agricultural practices. Riparian buffers were identified as a tool available to farmers and catchment managers to achieve water quality improvements. This study compares 10 m wide regenerating grass and Eucalyptus globulus buffer performance. Surface and subsurface water quality were monitored over a 3‐year period. Nutrient and sediment transport were both dominated by subsurface flow, in particular through the B‐horizon, and this may seriously limit the surface‐runoff‐related functions of the riparian buffers. Riparian buffer trapping efficiencies were variable on an event basis and annual basis. The grass buffer reduced total phosphorus, filterable reactive phosphorus, total nitrogen and suspended sediment loads from surface runoff by 50 to 60%. The E. globulus buffer was not as effective, and total load reductions in surface runoff ranged between 10 and 40%. A key difference between the grass and E. globulus buffers was the seasonality of sediment and nutrient transport. Surface runoff, and therefore sediment and nutrient transport, occurred throughout the year in the E. globulus buffer, but only during the winter in the grass buffer. As a consequence of high summer nutrient and sediment concentrations, half the annual loads moving via surface runoff pathways through the E. globulus buffer were transported during intense summer storms. This study demonstrates that grass and E. globulus riparian buffers receiving runoff from pasture under natural rainfall can reduce sediment and nutrient loads from surface runoff. However, in this environment the B‐horizon subsurface flow is the dominant flowpath for nutrient transport through the riparian buffers, and this subsurface flow pathway carries contaminant loads at least three times greater than surface runoff. Copyright © 2006 John Wiley & Sons, Ltd.     

黄河流域多时空尺度土地利用与水质的关系

为探究不同时空尺度土地利用与水质的相互关系,以黄河流域为研究对象,基于2018年河流水质与土地利用数据,提取3种空间尺度(1000 m河段缓冲区、500 m河岸带缓冲区、子流域)的景观格局指数,结合4个季节河流水质指标,采用冗余分析方法定量探讨土地利用对河流水质的多时空尺度影响,获得水质资源管理最有效的时空尺度.结果表明:(1)在3种空间尺度下,耕地和草地占比最大.斑块密度(PD)在1000 m河段缓冲区尺度下更大,香农多样性指数(SHDI)在3种空间尺度下变化不大;(2)水质指标季节性和空间性存在显著差异,夏季水质优于冬季;(3)土地利用在夏季和1000 m河段缓冲区尺度下对水质影响最显著;(4)不同景观格局指数对河流水质产生的影响不同,耕地面积占比和林地面积占比、未利用地面积占比和SHDI对水质影响最为显著,除了耕地面积占比和PD值的增加会导致水质下降外,其余土地利用指数在大多数时空尺度下均可改善水质.该研究基于不同的时空尺度对土地利用与水质的关系进行定量研究,为合理规划土地资源提供参考,从而保护流域水质安全.     

离散型湖泊水体提取方法精度对比分析

基于卫星遥感的陆地水体提取方法多种多样,并且应用广泛.对于水体分布支离破碎的枯水期湖泊,准确的水体提取方法尚不明晰,直接影响湖泊水域面积的提取精度.以鄱阳湖湖区为研究对象,利用ALOS遥感影像,以2.5 m高分辨率全色波段融合影像非监督分类(ISODATA)得到的水体面积为参考值,分别使用归一化水体指数(NDWI)法、NDWIISODATA法和基于近红外(NIR)的ISODATA法提取了10 m分辨率的水体分布,分析了不同方法提取结果之间的差异性及产生原因.结果表明:3种方法均可以较好地提取出水体,但利用ISODATA法提取的水体细部信息更为明显,面积值较NDWI法更大;相对于近红外单波段而言,基于NDWI图像的ISODATA法提取水体的精度更高.纵观3种方法,基于NDWI图像的ISODATA法提取的水体精度最高,基于近红外波段的ISODATA法提取结果次之,NDWI阈值法的提取效果最差.研究结果对于离散型湖泊水体提取方法及数据源的选择等具有重要的借鉴和参考意义.     

Zn2+浓度对固氮鱼腥藻(Anabaena azotica Ley)光能转化特性的影响

本实验对在不同Zn^2 浓度条件下培养的固氮鱼腥藻（Anabaena azotica Ley）的生长、光合放氧速率和叶绿素荧光参数Fv/Fm进行了测定。结果表明,当Zn^2 浓度为1.0μmol/L时,其比生长速率（Specifie growth rate）最大,光合放氧速率和Fv/Fm值最高,当Zn^2 浓度大于等于5.0μmol/L是会抑制A.azotica Ley的生长和光合作用,对在0μmol/L和5.0μmol/L Zn^2 发生反应的藻蓝蛋白溶液的可见光吸收光谱的分析,发现前者624nm处藻胆体的吸收峰和后者620nm处藻蓝蛋白的吸收峰都因Zn^2 的作用而下降,推测藻蓝蛋白为Zn^2 影响光合作用的位点之一,碳酸酐酶活性的测定表明Zn^2 的浓度水平会影响其活性大小,推测是Zn^2 影响光合作用的另一途径。     

微囊藻群体总RNA提取方法的比较

微囊藻群体富含多糖类物质是影响微囊藻RNA提取的关键因素.为了获得高质量的微囊藻群体总RNA,对比分析4种针对多糖含量较高的方法——方法 1 PGTX-bead法、方法 2 CTAB-bead法、方法 3 Fast RNAPro Blue Kit和方法 4RNeasy Mini Kit对微囊藻群体总RNA的提取效果.采用琼脂糖凝胶电泳检测微囊藻群体RNA的完整性,Nanodrop ND1000分光光度计检测RNA纯度及浓度,并采用q PCR检测DNA污染情况.结果表明,4种方法都能从微囊藻群体中提取获得RNA,并在去除DNA后都可以进行RT-PCR等后续实验.方法 1 PGTX-bead提取的RNA产量最高,纯度好,DNA污染小,成本低,适合从微囊藻群体中大量提取RNA;方法 2 CTAB-bead提取的RNA样品产量也较高,但DNA污染严重,适合需要同时提取DNA和RNA的样本;方法 3 Fast RNAPro Blue Kit和方法 4 RNeasy Mini Kit提取的RNA产量都较低,但方法 4操作简单,耗时短,所检测目的基因的相对表达量较高,更适合从少量的微囊藻群体中提取总RNA.     

Performance of grass and eucalyptus riparian buffers in a pasture catchment,Western Australia,part 1: riparian hydrology

Rainfall takes many flowpaths to reach a stream, and the success of riparian buffers in water quality management is significantly influenced by riparian hydrology. This paper presents results from hydrometric monitoring of riparian buffer hydrology in a pasture catchment. Runoff processes and riparian flowpaths were investigated on two planar hillslopes with regenerating grass and E. globulus buffers. Surface runoff and subsurface flows (A‐ and B‐horizons) were measured for 3 years using surface runoff collectors, subsurface troughs and piezometers. Water volumes moving through the riparian buffers via the measured flowpaths were ranked B‐horizon ? surface runoff ≈ A‐horizon. Runoff volumes through the B‐horizon troughs were an order of magnitude greater than those recorded for the most productive surface runoff plots or the A‐horizon troughs. Subsurface runoff and saturation‐excess overland flow (SOF) were limited to the winter months, whereas infiltration‐excess overland flow (IEOF) can occur all year round during intense storms. Surface runoff was recorded on 33 occasions, mostly during winter (late May–early October), and total annual surface runoff volumes collected by the 20 unconfined (2 m wide) runoff plots varied between > 80 and < 20 m³. Subsurface flow only occurred in winter, and the 6 m wide B‐horizon subsurface troughs flowed above 1 l s^?1 continuously, whereas the A‐horizon troughs flowed infrequently (<6 days per year). In summer, surface runoff occurred as IEOF during intense storms in the E. globulus buffer, but not in the grass buffer. Observations suggest that surface crusting reduced the soil's infiltration capacity in the E. globulus buffer. During winter, SOF and seepage were observed in both buffers, but subsurface flow through the B‐horizon was the dominant flowpath. Key hydrologic differences between the grass and tree buffers are the generation of IEOF in the E. globulus buffer during intense summer storms, and the smaller subsurface runoff volumes and fewer flow days in the E. globulus buffer. Low surface runoff volumes are likely to limit the potential of these buffers to filter pollutants from surface runoff. High subsurface flow volumes and saturated conductivities are also likely to limit the residence time of water in the subsurface domain. Based on their hydrologic performance, the key roles of riparian buffers in this landscape are likely to be displacing sediment and nutrient‐generating activities away from streams and stabilizing channel morphology. Copyright © 2006 John Wiley & Sons, Ltd.     

Preliminary Interlaboratory Trial for ISO/DIS 12010: Determination of Short Chain Polychlorinated Alkanes (SCCP) in Water

A first preliminary interlaboratory trial was planned to prepare ISO/DIS 12010: Water quality – determination of short chain polychlorinated alkanes (SCCP) in water – method using GC/MS and electron capture negative ionisation (ECNI). The task was to determine the sum of short chain polychlorinated n‐alkanes with carbon chain lengths of C₁₀–C₁₃ and a chlorine content between 49 and 67% in water by GC‐ECNI‐MS and quantification by multiple linear regression described in ISO/DIS 12010 as the compulsory method. Distributed samples were obtained from a real water extract spiked with a target concentration of 0.4 µg/mL sum of SCCP, i.e. the environmental quality target level according to the Water Framework Directive. The interlaboratory trial included the calibration, a column chromatographic clean up, a concentration step and an integration of chromatographic unresolved humps as well as the quantification with multiple linear regression. Reproducibility standard deviations between 21.5 and 22.9% were achieved by 17 participating laboratories from four countries. The method outlined no significant difference of the results between the standard solution and a real water matrix extract. On the basis of this succeeded preliminary interlaboratory trial the final interlaboratory trial for validation of ISO 12010 was prepared in autumn 2010.