藻类的采集和鉴定课件

IdentificationandCollectionofAlgae

胡韧

暨南大学水生生物研究中心SampleCollectionMonitoringSiteSelection1MonitoringTimeSelection2RequiredEquipment3Collectionmethods4Instituteofhydrobiology,JinanUniversityMonitoringSiteSelection11Segmentsthathaveverydifferentwaterqualityorpollutionpotentialinspecificareasshouldhavemorethanonestation.2Selectmonitoringsitesthatbestrepresentwaterqualityconditionsofawaterbody.3Sitesmaybelocatedbelowmajorpollutionsources.Instituteofhydrobiology,JinanUniversityMonitoringSiteSelection24Lakesandestuariesshouldhavestationsinthemajorarmsandnearthedam(reservoirs).5Forroutinemonitoringinfreshwaterstreamsandrivers,sitesmusthaveeitherastreamflowgageorbesuitableforconductingflowmeasurement.6Asageneralrule,streamsbetween25and50mileslongrequiretwostations.Instituteofhydrobiology,JinanUniversityMonitoringTimeSelection1-4times/monthSamplingfrequencyAccordingtodifferentprogramSummerseasonalSpringSummerInstituteofhydrobiology,JinanUniversityMonitoringTimeSelectionFlowBaselinemonitoringincludessamplescollectedoverarangeofflowconditions.FlowWhensamplesarecollectedduringabnormallyhighorlowflow,theabnormalconditionswillberecordedinthefieldlogbookandontheobservationlinesofthesampledataform.

Instituteofhydrobiology,JinanUniversity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6μm或10μm的浮游生物网过滤获得。RequiredEquipmentInstituteofhydrobiology,JinanUniversity边长1M45cm9cm63cm自制浮游植物网Instituteofhydrobiology,JinanUniversity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，应取至透明度的1.5倍且包括斜温层的深度。Instituteofhydrobiology,JinanUniversity其它定量采样工具intergratedhose-pipesampler-5mlengthof2.5cmdiameterplasticpipingwithaweightedcollaratoneendacordattachetothehoseandboatarubbercorktofitoneendofthehoseInstituteofhydrobiology,JinanUniversity其它采样用品透明度盘浊度计pH计多参数水质仪Instituteofhydrobiology,JinanUniversity透明度测定Procedure◘Lowerdiskintothewaterontheshadedsideoftheboatwhilekeepingafirmgripontheline.◘Asthediskislowered,countthenumberofdepthmarksontheline.◘Keeploweringthediskuntilitisnolongervisible;recordthedepthaccordingthelinemarks.◘Slowlyraisethedisk,andwhenitappearsagain,recordthedepth.◘TheaverageofthesetwodepthsistheSecchidepth.Instituteofhydrobiology,JinanUniversityFieldDataLogbooksStationID

SamplingDate

Location

SamplingDepth(ifotherthansurface)

SamplingTime

SampleCollector'sInitials(ifseveralpersonsintheregioncollectthisdata)

Recordofallmeasuredfieldparametersandtheirrespectivevalues.Foreachvisittoanindividualstationwherefieldand/orsamplesarecollectedrecordthefollowing:Instituteofhydrobiology,JinanUniversityFieldphysicochemicalparametersincludepartorallofthefollowing:

DissolvedOxygen

ChlorineResidual

Temperature

Salinity(tidalwatersonly)

pH

SecchiDiskTransparency

SpecificConductance

DaysSinceLastPrecipitationEvent(significantenoughtoinfluencewaterquality).

FlowSeverityInstituteofhydrobiology,JinanUniversityFieldDataLogbooksInstituteofhydrobiology,JinanUniversityOtherObservationsWaterAppearanceColor,unusualamountofsuspendedmatter,debrisorfoam,etc.WeatherRecentmeteorologicaleventsthatmayhaveimpactedwaterquality;heavyrains,coldfront,verydry,verywet,etc.BiologicalActivityExcessivemacrophyte,phytoplanktonorperiphytongrowth,Theobservationofwatercolorandexcessivealgalgrowthisveryimportantinexplaininghighchlorophyllavalues.Otherobservationssuchasfish,birdsandspawningfisharenoted.Instituteofhydrobiology,JinanUniversityOtherObservationsUnusualOdorsHydrogensulfideodor,mustyodor,sewageodor,etc.WatershedorInstreamActivitiesInstreamordrainagebasinactivitiesoreventsthatareimpactingwaterquality;bridgeconstruction,shorelinemowing,livestockwateringupstream,etc.Instituteofhydrobiology,JinanUniversityOtherObservationsRecordofPertinentObservationsRelatedtoWaterQualityandStreamUses.Ifthewaterqualityconditionsareexceptionallypoor,notethatstandardsarenotmetintheobservations,forexample,dissolvedoxygenisbelowminimumcriteria.Uses-swimming,wading,boating,fishing,irrigationpumps,navigation,etc.Eventually,forsettingwaterqualitystandards,thelevelofusewillbebasedoncommentsrelatedtotheleveloffishingandswimmingactivitiesobservedatastation.Instituteofhydrobiology,JinanUniversitySampleFixation固定液的配方如下：定性样品固定液甲醛：丙三醇：水=10：10：80（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。Instituteofhydrobiology,JinanUniversity从10L混合水样中取1L作为定量样品，用Lugol’s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。Instituteofhydrobiology,JinanUniversitySedimentationandConcentration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1μm～50μm之间，经碘液固定后，下沉较快，所以静置时间一般可为48h。Instituteofhydrobiology,JinanUniversity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20μm的筛绢封盖。Instituteofhydrobiology,JinanUniversity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2－3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parmarfilm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Instituteofhydrobiology,JinanUniversityIdentifyandCount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。浮游植物计数框：由玻璃条组成的方框，面积为20mm×20mm，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。Instituteofhydrobiology,JinanUniversityCountMethods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100~500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。Instituteofhydrobiology,JinanUniversityCalculation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/mLA——计数框面积，mm2

Ac——计数面积或视野面积，mm2Vs——1L原水样沉淀浓缩后的体积，mL

Va——计数框的容量，mLn——