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1、Identification and Collection of Algae胡韧暨南大学水生生物研究中心,Institute of hydrobiology,Jinan University,Sample Collection,Institute of hydrobiology,Jinan University,Monitoring Site Selection 1,Institute of hydrobiology,Jinan University,Monitoring Site Selection 2,Institute of hydrobiology,Jinan University,M
2、onitoring Time Selection,Institute of hydrobiology,Jinan University,Monitoring Time Selection,Institute of hydrobiology,Jinan University,Monitoring Time Selection,Institute of hydrobiology,Jinan University,Required Equipment,浮游生物采集网 25号（64 m）13号（86 m）采水器 2.5 L 5 L,Institute of hydrobiology,Jinan Uni
3、versity,定量采样方法：用采水器取水面以下0.5m 至离湖底0.5m 的混合水样10L。,定性样品用孔径为6m 或10m的浮游生物网过滤获得。,Required Equipment,Institute of hydrobiology,Jinan University,自制浮游植物网,Institute of hydrobiology,Jinan University,不同水体定量样品的采集,对于一般富营养化调查，取水面以下0.5米处的水样即可。对于多样性调查，则要取生物多样性更高的地方，所以多采取混合水样。没有分层的浅水湖泊，每1米一个样取至水底上方0.5米处。有分层的水体，应取至透明度
4、的1.5倍且包括斜温层的深度。,Institute of hydrobiology,Jinan University,其它定量采样工具,intergrated hose-pipe sampler-5 m length of 2.5 cm diameter plastic piping with a weighted collar at one enda cord attache to the hose and boata rubber cork to fit one end of the hose,Institute of hydrobiology,Jinan University,其它采样用
5、品,透明度盘浊度计pH计多参数水质仪,Institute of hydrobiology,Jinan University,透明度测定,Procedure Lower disk into the water on the shaded side of the boat while keeping a firm grip on the line.As the disk is lowered,count the number of depth marks on the line.Keep lowering the disk until it is no longer visible;record
6、the depth according the line marks.Slowly raise the disk,and when it appears again,record the depth.The average of these two depths is the Secchi depth.,Institute of hydrobiology,Jinan University,Field Data Logbooks,Station ID Sampling Date Location Sampling Depth(if other than surface)Sampling Time
7、 Sample Collectors Initials(if several persons in the region collect this data)Record of all measured field parameters and their respective values.,For each visit to an individual station where field and/or samples are collected record the following:,Institute of hydrobiology,Jinan University,Field
8、physicochemical parameters include part or all of the following:,Dissolved Oxygen Chlorine Residual Temperature Salinity(tidal waters only)pH Secchi Disk Transparency Specific Conductance Days Since Last Precipitation Event(significant enough to influence water quality).Flow Severity,Institute of hy
9、drobiology,Jinan University,Field Data Logbooks,Institute of hydrobiology,Jinan University,Other Observations,Water Appearance Color,unusual amount of suspended matter,debris or foam,etc.Weather Recent meteorological events that may have impacted water quality;heavy rains,cold front,very dry,very we
10、t,etc.Biological Activity Excessive macrophyte,phytoplankton or periphyton growth,The observation of water color and excessive algal growth is very important in explaining high chlorophyll a values.Other observations such as fish,birds and spawning fish are noted.,Institute of hydrobiology,Jinan Uni
11、versity,Other Observations,Unusual Odors Hydrogen sulfide odor,musty odor,sewage odor,etc.Watershed or Instream Activities Instream or drainage basin activities or events that are impacting water quality;bridge construction,shoreline mowing,livestock watering upstream,etc.,Institute of hydrobiology,
12、Jinan University,Other Observations,Record of Pertinent Observations Related to Water Quality and Stream Uses.If the water quality conditions are exceptionally poor,note that standards are not met in the observations,for example,dissolved oxygen is below minimum criteria.Uses-swimming,wading,boating
13、,fishing,irrigation pumps,navigation,etc.Eventually,for setting water quality standards,the level of use will be based on comments related to the level of fishing and swimming activities observed at a station.,Institute of hydrobiology,Jinan University,Sample Fixation,固定液的配方如下：定性样品固定液 甲醛：丙三醇：水=10：10
14、：80（体积比），可再加1ml冰醋酸，可以防止易收缩的藻类变形。定量样品固定液（鲁哥试液）20g碘化钾溶解于含有20ml冰醋酸的200ml的水中，再加10g碘溶解其中，用棕色瓶保存。改良的可加4ml丙三醇，可用于对浮游动物的保存。,Institute of hydrobiology,Jinan University,从10L混合水样中取1L作为 定量样品，用Lugols 碘液固定并沉淀浓缩至50mL，作定量计数用。将部分样品经酸处理，用于在油镜下对硅藻进行鉴定。,Institute of hydrobiology,Jinan University,Sedimentation and Conce
15、ntration,沉淀和浓缩在筒形分液漏斗中进行。把分液漏斗固定在架子中，放在稳定的桌子上。将水样倒入分液漏斗，使浮游植物自然沉淀。理论上，浮游植物最小颗粒沉降速率为0.3cm/h,若分液漏斗中水柱高度为20cm，则需沉淀60h。实际上，藻类大小在1m50 m之间，经碘液固定后，下沉较快，所以静置时间一般可为48h。,Institute of hydrobiology,Jinan University,分级沉淀法,没有分液漏斗时，也可采取分级沉淀法。沉淀后，用细小的玻璃管（直径小于2mm）借虹吸方法慢慢吸去上清液，注意不能搅动或吸出浮在表面和沉淀的藻类，虹吸管在水样的一端可用孔径为20 m的筛
16、绢封盖。,Institute of hydrobiology,Jinan University,分级沉淀法,将剩余的藻液（约20mL）放入容积为50mL的试剂瓶中。试剂瓶事先在30mL处做好标记。用吸出的上层清液冲洗分液漏斗23次，一起放入试剂瓶中，定容到30mL。需要长期保存的样品，应加入少许甲醛溶液，并用石蜡或parmar film膜封口。样品瓶上应写明采样日期，地点，采样体积和浓缩体积等,Institute of hydrobiology,Jinan University,Identify and Count,对采到的优势种要求鉴定到种，一般到属。疑难种类要保存标本以备进一步鉴定。浮游植
17、物计数框：由玻璃条组成的方框，面积为20mm20mm，容量为0.1mL，框内划分为横直各10行格，共100个小方格。将计数样品充分摇匀后，迅速吸取0.1mL样品至计数框中，盖上盖玻片。计数框内应无气泡，也不应有样品溢出。气温高时，为了防止在长时间计数过程中水分蒸发而出现气泡，可在盖玻片四周封以液体石蜡。计数时，显微镜的目镜可用10，物镜用40，但根据情况可加以变动。,Institute of hydrobiology,Jinan University,Count Methods,行格法：对计数框上的第二行、五行和八行，三行共30格小方格进行计数，现已较少采用。视野法：利用显微镜的目镜视野来选取
18、计数的面积。先用台微尺量得在一定放大倍数下的视野直径，然后按圆面积公式求得视野面积。应利用计数框上的方格或显微镜机械移动台上的标尺刻度，使选取的视野在计数框上均匀分布。计数的视野数目应根据样品中浮游植物数量的多少确定。一般计数100500个视野，使所得的浮游植物计数值至少在300以上。可以先计数100个视野，如计数后数值太少，再增加100个，依此类推，计数的视野应均匀分布在计算框的全部面积内。,为减少工作量，一般不对整个计数框内水样中的浮游植物都计数，而只选取其中一部分样品计数。选取过程是一个次级抽样过程，故要考虑抽样的大小和代表性，常用行格法和视野法。,Institute of hydrob
19、iology,Jinan University,Calculation,把计数结果换算为原来所采的水样中浮游植物数量时，用下列计算公式：,式中：N浮游植物数量，cell/mL A计数框面积，mm2 Ac计数面积或视野面积，mm2 Vs 1L原水样沉淀浓缩后的体积，mL Va计数框的容量，mL n 计数所得的浮游植物数目，cell,Institute of hydrobiology,Jinan University,Note,采用某种计数方法后，不要随意改变，以保证结果的可比性。若遇到一个浮游植物个体或细胞的一部分在行格或视野内，另一部分在行格或视野外，则可自行规定。如：在行格上线或视野上半圈的
20、个体或细胞不计数，而在下线或下半圈的细胞计数。计数前应对样品做定性观察，以熟悉主要种类及其形态特点。计数时应把注意力集中到主要种类，对数量极少的稀有种类，一时确定不了归属的，可先计数，需要时再鉴定种类。当某一个或几个优势种的数目非常多时，可用计数器，对其单独计数。必要时先统计这些种类的数目，再看其它种类。,Institute of hydrobiology,Jinan University,对于比较难判断细胞数的群体，则任选20 个个体在高倍下观察，测出细胞数，取平均值。由于硅藻在直接观察时无法定种，所以计数时先计算总的硅藻细胞数，然后将部分样品酸处理后在油镜下定种并计算出各种硅藻间细胞数的比例。,Institute of hydrobiology,Jinan University,群体和丝状体的计算方法,对丝状体和一些较小的群体，可以先计算个体数，然后求出该种类的个体的平均细胞数，进行换算。对形成“水华”的优势种类，如：微囊藻，计数前可用加碱、加热、超声波振荡等方法使其散开成为单个细胞或少数细胞的群体。,计数的单位可以用个体或细胞表示。用个体数表示，计数时较省力，但由于藻体有的是单细胞，有的是数目相差悬殊的细胞组成的群体，因此用个体数表示不及用细胞数表示精确。,
