sampling guide for the collection of … · dragonfly mercury project – sampling guide scope and...
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U.S.GeologicalSurvey(USGS)ForestandRangelandEcosystemScienceCenterUniversityofMaine(UMaine)–SchoolofForestResources
SchoodicInstitute–AcadiaLearningProgramNationalParkService(NPS)–AirResourcesDivision(ARD)
DRAGONFLYMERCURYPROJECT
SAMPLINGGUIDEFORTHECOLLECTIONOFDRAGONFLYLARVAESAMPLESFROMNATIONALPARKSFORMERCURYANALYSIS
Preparedby:SarahJ.Nelson,AssociateResearchProfessor,UniversityofMaine
Phone:207-581-3454|Email:[email protected]
CollinEagles-Smith,SupervisoryResearchEcologist,USGS-FRESCPhone:541-750-0949|Email:[email protected]
JamesWillacker,Ecologist,USGS-FRESC
Phone:541-750-0957|Email:[email protected]
Editedby:ColleenFlanaganPritz,Ecologist,NPS-ARD
Phone:303-969-2806|Email:[email protected]
AmandaKlemmer,AssistantResearchProfessor,UniversityofMainePhone:207-581-4513|Email:[email protected]
Contributor:AndreaBlakesley,EnvironmentalSpecialist,DenaliNationalPark&Preserve
Phone:907-683-9545|Email:[email protected]
RevisedMay2017
DragonflyMercuryProject–SamplingGuide
TableofContentsScopeandApplication.............................................................................................................2Overview......................................................................................................................................41. Prepare..........................................................................................................................52. Collectsamples...........................................................................................................63. Identify..........................................................................................................................84. Choose,Measure,&Bag...........................................................................................85. Store&Send..............................................................................................................116. Usethedata!..............................................................................................................11AppendixA.Nets.....................................................................................................................12AppendixB.Howcleanis“clean”?....................................................................................13AppendixC.ResearchPermitApplication–exampleRPRStext............................14AppendixD.HowtouseiNaturalist..................................................................................15AppendixE.SacrificingDragonflyLarvaeforScience................................................19
ParkscientistsandstudentsfromArizonaCollegePrepAcademy searchfordragonflylarvaeinSaguaroNationalPark,AZ,2015.Photo:DanBell.
DragonflyMercuryProject–SamplingGuide
ScopeandApplicationBackground
Mercury(Hg)isagloballydistributedcontaminantthatcanharmhumanandwildlifehealth.Initstoxicmethylatedform,mercurybioaccumulates(buildsup)inaquaticandterrestrialfoodwebs.Effectscanincludereproductiveandneurologicalimpairment.Dueinparttoemissionsfromcoal-burningpowerplants,evenremotenationalparkenvironmentsreceivemercurydepositionfromtheatmosphere.(Seehttp://www.nature.nps.gov/air/AQBasics/mercury.cfmforbackgroundonmercuryinnationalparks.)
MercurythreatensnaturalresourcestheNationalParkServiceischargedwithprotecting.Thiscitizenscienceprojectencouragesstudentsandvisitorsinnationalparkstocollectdragonflylarvaeformercuryanalyses.Dragonflylarvae(Odonata:anisoptera)couldserveasindicatorsofecosystemhealthbycharacterizingtheriskandpotentialtoxiceffectsofmercury.Theseaquaticmacroinvertebratesarelong-lived(upto5yearsaslarvae)beforeemergingasadultdragonflies,widespreadacrosstheU.S.,predatory(i.e.,pronetohigherconcentrationsofmercury),importantpreyforfishspecies,andtheyreflectthemercurysensitivityofaspecificwatershed.Moreover,theyarerelativelyeasytocollect!
TheDragonflyMercuryProject(DMP)connectspeopletoparks,advancestheNPSeducationalmission,fostersbiodiversitydiscoveryopportunities,andprovidesbaselinedatatobetterunderstandthespatialdistributionofmercurycontaminationinnationalparks.TheDMPexpandsthegeographicscopeofresearchpreviouslyconductedbyscientistsandcitizenscientists,andprovidesdatathatcanbecomparedacrossparks.Earlydataarebeingusedtodevelophypothesesregardingwhethermercuryvarieswithodonatelarvalbodysizeorbyfamily,orwhetherasite’slandscapesettingdrivesvariabilityinmercuryinodonatelarvae.Seeprojectwebpage,availableathttp://www.nature.nps.gov/air/Studies/air_toxics/dragonfly/index.cfm.EducationalcontentandlessonplansforusebyteachersandNPSstaffarealsoavailable,(seehttp://www.nature.nps.gov/air/studies/air_toxics/dragonfly/educational.cfmandhttp://participatoryscience.org/)ultimatelyhelpingtoraiseawarenessaboutmercuryimpacts.TheDMPenlightensanewgenerationofcitizenscientistsabouttheconnectionofalllivingthingsandtheinfluencehumanshaveuponnaturalsystems,andhowenvironmentally-responsibledecisionscanprotectourparksandtheplanet.
Thecitizenscienceefforttocollectdragonflylarvaefromnationalparksformercuryanalysisstartedin2011withfourNPSunitsintheNortheastandGreatSmokyMountains,expandedin2012toinclude11parks,andin2013toinclude23parksacrossallsevenNPSregions:Northeast,Southeast,NationalCapitol,Midwest,Intermountain,PacificWest,andAlaska.Theprojectcontinuedtogrowfrom33parksin2014to49parksfromaroundthenationin2015.In2016,theNationalParkServiceCentennial,59parksengagedintheDragonflyMercuryProject.Summarynumbersfrom2014-2016provideatotalof224sitesfrom77parkssampled,andover7,500dragonfliesanalyzedformercury.Over3,000citizenscientistshavebeenengaged.In2017,wearecontinuingtheeffortbysamplingsomenewparksandsupportingtheresamplingofsomeparksthathavealreadyparticipated.Citizenscientistsinvolvedincludestudentsrangingfromelementaryschooltocollege-aged,parkvisitors,andBioBlitzparticipants.
DragonflyMercuryProject–SamplingGuide
StudyDesignTheoptimalstudydesignis3sitesperparkthatrepresentagradientinmercury-relevantchemistry;oragradientinlandscapeconditions,suchasahighandlowelevation,amountofwetlandsadjacenttoorupstreamfromthesite,orforestedversusurbanlandcover.Considerscopingthesamplinglocationsinadvanceofcollectingsamples.Doestheproposedsamplinglocationhaveahealthypopulationofdragonflies?(Asamplesizeof15dragonflylarvaepersiteispreferred.)Willnearbyriparianfloraandfaunagettrampled?Isthesitespaciousenoughforagroup?
ResearchPermitAparkresearchpermitwillbeneededtoconductthisproject.Inaddition,insomecases,astatepermitmaybeneededtocollectdragonflylarvae.ExampletextforthepermitapplicationcanbefoundinAppendixAofthissamplingprotocol.TheDMP’sfactsheetandothermaterialswillprovidefurtherassistance.(SeeURL’sabove).
SafetyFieldworkcarriessomeinherentrisks,andthesafetyofstudyparticipantsisaprimaryconcern.Thevariedandspectacularresourcesofournationalparks,andtheengagementofcitizenscientistswhoarepotentiallylessskilledinfieldworkandtheassociatedhazards,presentachallengetosafelyconductthisproject.Further,travelinremoteareasmayberequiredtoconductsciencethathasmeaningfulinferencetotheparks’diverselandscapes,increasingrisks.BeattentiveofthepossiblerisksandrefertoadetailedSafetyStandardOperatingProcedure(SOP)ortoaJobHazardAnalysis(JHA)thatisdevelopedspecifictosurfacewatersampling,backcountrywork,travelinginhighelevations,poisonivy,tickbornedisease,etc.([email protected].)Riparianedgescanbeveryslippery,asarestreamandlakebottoms.Highwaterlevelsandturbulentflowsmaycauseanindividualtolosebalanceinthewater.Collectivelydefine,communicate,andenforceyourpark’ssafetyphilosophy,standards,andguidelines;followthetenetsofOperationalLeadership(e.g.,briefyourteampriortosamplingonmissionelements:Communication,ContingencyResources,Environment,Planning,TaskComplexity,Supervision,TeamFitness,andTeamSelection)andotherrelevantsafetytrainingssuchasswiftwaterrescue;allowsufficienttimeforoversightandinteractionwithyourgroup;andfollowDepartmentalguidelinesontheuseofpersonalprotectiveequipment,suchaspersonalflotationdevices(PFDs).WehighlyrecommendcitizenscientistsusePFDswhensamplingANYwaterbodies,andinfact,USGSscientistsarerequiredtowearPFDs.Nothinginthisprogram’smission,projectgoals,orday-to-dayobjectivesissoimportantastocompromisethehealthandwell-beingofthoseparticipatinginthisproject.
DragonflyMercuryProject–SamplingGuide
OverviewThisprotocolhasseveralsections.Eachsamplingprocedureisfairlystraightforward,afteryou’vegottenthehangofit.Youshouldreadeachsection’sspecificinstructionsonthefollowingpages.Thegeneralstructureisasfollows:
1. Preparebygatheringfieldgear.Somegearisprovidedinthesamplingkit,butyou’ll
needtoroundupafewitems.Checkwhat’sneededseveralweeksbeforesampling(incaseyouneedtoordersomething).
2. Collect!
First,collectdragonflylarvaesamples.Manyhandssamplingmakesthisgoquicker.Larvaearecollectedusingnetsandtemporarilystoredinasmalltoteordishpanwithlakewateruntilbeingsorted.Onceatleast10larvae(butideally15)havebeencollected,samplesaresorted,andupto15individualsareselectedformercuryanalysis.Thesearemeasured,identifiedtofamily,thenindividuallybaggedtopreventcontamination.
Second,observationsaboutthesite.Manyarequalitative/visual,buttheyarestillveryusefuldata.Youknowyoursitebestandyoucanhelptheprojectscientistsunderstandyourwatershed.WealsoencourageobservationsthroughiNaturalist.org.
3. Shipyoursamples.You’llshipdragonflysamplesovernightinacoolerwithdryice.Dragonfliesshouldgoinafreezerafterthesamplingtripisover.Theycanstayinyourfreezerforseveralweeks,ifyouhavemultiplesitestosampleoverextendedtimeperiods.
DragonflyMercuryProject–SamplingGuide
1. PrepareThefollowingsamplingsuppliesareneededtocollectdragonflylarvaesamplesfromeachsite.Allsupplieswillbeprovidedinthesamplingkit,withtheexceptionofthosematerialsinbold.Participatingparksmustprovidethoseitemsseparately.
MaterialsDragonflysampling:
! Nets*–DNetsordipnets! Clean,whitedishpan,bucket,oricecubetrays! Clean,newplasticspoons
! Brandnewzipper-sealbags–30smallsize+3largesize! PowderfreeNitrilegloves(blueorpurplecolor)! Aplasticrulerwithmmscale–clearisbest! Tagsforouterbag,pre-printedwithIDcodes
Generalfieldsupplies:! Sharpiemarkerandpencil/pen! Fieldsheets! Onecooler! Trashbag! Optional–handlens,GPS,camera,macroinvertebratefield
guide,waders! TheparkorcitizensciencegroupmustprovidePFDs
(personalflotationdevices)
Shipping:! Dryicefordragonflies! Dryicelabelandpre-paidshippinglabelsforreturnshipping
Resources:• Seetheprojectwebsiteforeducationaltools,informationabout
mercuryanddragonflies,curriculumactivities,andnewsaboutprevioussamplingefforts.
• Watchthetrainingvideosavailableat:http://www.schoodicinstitute.org/what-we-offer/educational-scientific-partnerships/dragonfly-mercury-project/
• Follow“Six-LeggedScoutsintheNationalParks”onFacebookforupdatesandnews.*MoreinformationonnetsinAppendixA.
DragonflyMercuryProject–SamplingGuide
2. Collectsamples
• Fill-outthesiteinformation(includingpark,sitename,sitecoordinates,date,andleadcollector’scontactinformation)onbothfielddatasheets.Ifthisinformationisnotprovidedonthedatasheetsyouwillbecontactedbytheprojectcoordinatortogettheinformation.
• Collectdragonflylarvae.• Repeatthesamplingfor3waterbodiesperparkwhenavailable.• Wehighlyencourageyoutotakephotosofthesamplingsitesandofthecollectionefforts.
ImagesofengagedNPSemployees(inuniform)withcitizenscientists(wearingPFDsifsampling)areespeciallypreferred.WecannotusephotosofparticipantsinornearwaterwithoutaPFDon.
• TakeobservationsusingiNaturalist(ifusing;seeAppendixDforinstructions).• ReferenceAppendixEon“SacrificingDragonflyLarvaeforScience”inordertoaddresspotential
ethicalconcernsraisedbyyourteamofcitizenscientists.
Procedure1. Lookaround.Locatelikelyhabitatsfordragonflies:vegetatedbankmargins,snagsandlogs,
aquaticvegetationanddecayingorganicmatter,silt/sand/gravelsubstrate.2. Noteobservationsandfinishcompletingthefieldsheet.Takeasitephoto!3. Rinsethenetsanddishpanswithwaterfromthesiteyouaresampling.4. Partlyfilldishpanorbucketwithwaterfromthesite.Icecubetraysarenicesorting
containerstoo–theycanbefilledwithstream/pondwateraswell.5. Jabyournetintothelikelyhabitats.Startdownstreamfirst,andworkupstreamifinflowing
water.Jabafewtimesthensweepthenetuptothesurface.Emptythenetintothedishpanorbucket–eitherbytippingitintothebucketorplucking(withaNitrile-glovedhandoraplasticspoon)larvaedirectlyfromthenet.Spendafewminutesjabbingandemptyingthenet.Ifyoudon’tfindmuchafterafewminutes,movetoanotherareawithinthesamewaterbody.Seeavideodemonstrationofcollectingdragonflylarvae:https://youtu.be/YXXCOU2MxHg
6. Ifyourteamconsistsofmultiplehands,recorddragonflyspecimenswithiNaturalist(App.D).7. Disinfectwaders,nets,andanyothergearthathasbeeninthewaterbeforemovingtoyour
nextsite.Consultyourpermitsincesomeparkshavedifferentprotocolsfordisinfection.Most(butnotall)willspecifya5%or10%bleachsolution,whichisamixtureofhouseholdbleachandwater,orsomeotherdisinfectingsolution.
submitted(atGreatSmokyMountainsNationalPark,NC/TN).
CitizenscientistssearchfordragonflylarvaeatLewis&ClarkNationalHistoricalPark,OR.
DragonflyMercuryProject–SamplingGuide
Procedures–CollectingDragonflyLarvaeSamples
KeepitClean!DragonflyLarvaeSamplingProtocolIt’seasytocontaminatesampleswithmercurybecauseit’sallaroundus:inourhair,insoil,leaves,etc.Soweneedtokeepeverythingultra---clean.
• Donottouchthesamples(individualdragonflylarvae)withanythingexceptacleanplasticspoonorpowder-freenitrileglove(purplegloves).
• Nevertouchtheinsideoftheinnerbag–thefirstbagyouputthesampleinto.• Thinkofeachindividualsampleasafreshstart–everythingneedstobecleanbetweensamplesandbetreatedjustlikethefirstsampleyoutook.
• Workinpairsandusethe“CleanHands–DirtyHands”method:1. Onepersonisinchargeofthesamplebags(“DirtyHands”)andtheotherwillpickup
andplacethesamplesinthebags(“CleanHands”).2. CleanHandsputsonaglove(oneglovedhandshouldsufficetopickandbaglarvae).
Oncethegloveison,CleanHandstouchesnothingbutthesampleandanypre-cleanedsupplies.DirtyHandsdoesnotneedtoweargloves.
3. Choose,measure,&bagupto15individualsperwaterbody/site.4. Ifyourteamconsistsofmultiplehands,recordspecimensusingiNaturalist(App.D).5. Usinganitrile-glovedhand,orplasticspoon,CleanHandsplacesonedragonflyintoa
samplebagwhileDirtyHandsholdsthebag.Nothingshouldtouchtheinsideofthebagexceptthedragonflylarva.CleanHandssealsthebag.
6. DirtyHandsusestheplasticruler,helduptotheindividualinthebag,andmeasuresthebodylength inmillimeters(mm)– fromthe frontofheadtoendof tailspine.Mostlarvaewillrangebetweenabout10–40mm,asareference.
7. DirtyHandsfillsoutatagforthebaggedindividualandslipsitandtheinnerbagintotheouterbag.YoursampleshaveanIDcodepre-printedonthelabels.Usethesecodesto record the lengths on the field sheet and to refer to a particular sample in yournotes.
8. DirtyHandssealstheouterbag.9. Be‘clean’withyourspoonand/orgloves.Iftheyhavebeentossedonthegroundor
contaminated–say,bybrushingbackyourhair–startwithafreshone.Swishyourspoonorglovedhandsinpondwater(inthedishpan)betweensamplestoensurethere’snocarryoverfromonesampletothenext.
10. Gotothenextindividualandrepeat.Placeallofthesmalldoublebagsintooneormoreofthelargerzippersealbagsforstorageandtokeepthemsafeandtogether.(SeeSection:4.Choose,Measure,&Bag)
11. Gentlyreturnextrasitewaterandinvertebratestothesamplesite.12. Packoutalltrash,disinfectgear.
DragonflyMercuryProject–SamplingGuide
3. IdentifyPleaseidentifydragonflylarvaetofamilyifyouarecomfortabledoingso.Thereareonly6majorfamiliesinmostplaces(seefollowingpages).Mostidentificationscanbeaccomplishedinthefieldwithahandlensandguidebook,oryoucanbagsamples(seeSectionIV)andbringthemtoacleanindoorworkspacetolookmoreclosely.Theonlinevideo“IdentifyingMaineDragonflyLarvaetoFamily”(http://vimeo.com/76713446)willalsoassistwithidentificationbecausesimilarlarvaefamiliesarefoundoutsideofMaine.
Identifyingtofamilyrequirescarefulinspection.TheIDwillbecheckedoncethelaboratoryreceivessamples,andsomesampleswillbefurtheridentifiedtospeciesthroughgeneticbarcodingorataxonomist,butidentifyingsampleswhentheyarefreshishelpfulintheeventofdamageduringshipping.
A. ItishelpfultodistinguishbetweenthetwoSuborderlevels:Dragonflies(Anisoptera)vs.Damselflies
(Zygoptera).Althoughbotharepredators,theAnisopterabodiesaremorerobustandmaythereforeconsumeandstoremoremercury.Anisopteraalsotendtohavelongeraquaticlifespans(upto5years)thanZygoptera(usually1year).
B. Ittakesspecializedtrainingtoclassifydragonflylarvaetothespecieslevel;wewilldothisforsomeindividualsinthelabandprovidetheidentificationinformationtoyoualongwiththemercuryandotherdata.Thisinformationisespeciallyusefulforbiodiversitydiscoveryactivities!
4. Choose,Measure,&Bag• Lookatthesamplesyou’vecollected.Ifyouhaveidentifiedtofamilyalready,pleasechooseat
least3individualsineachofthefamiliesyoufound.Ifyouonlyfoundoneindividualinacertainfamily,pleasedon’tsubmitthatone.Ideally,atleast10andupto15individuallarvaewillbeselectedpersite.Wewouldliketoanalyzesamplesfromacrosstherangeofsizespresentatasite,however,larvaelargerthan15mmareeasiesttomeasuremercuryin–pleasechoosemediumtolargersamplesoververysmalloneswhenpossible.
• Giventheriskofcontamination,itisveryimportanttofollowtheKeepitClean!Protocol(above)whilechoosing,measuring,&baggingdragonflylarvaesamples,andthroughthewholesamplingouting.
• Insum,oncesampleshavebeencollected,identified,andselected,eachdragonflylarvaeisdouble-bagged;placedfirstinaninnerzipperbag,thenwithlabelplacedinanouterbag,andstoredonwetice.
• UseiNaturalisttorecordeachspecimencollected.(SeeAppendixDforinstructions.)Ifyouaretakingaphotoofthespecimen,considerusinganiPhone/Androidmacrolens,ifavailable,anddoingsojustbeforethelarvaisbaggedtoobtaintheclearestpicturepossible!
DragonflyMercuryProject–SamplingGuide
Aeshnidae
Macromiidae
Notevarietyofbodytypes
Photosofexuviae:A.Anderson,KingfisherPhotography,OldTown,ME
CordulegastridaeLabiumextended
CorduliidaeEpithecaprinceps Labiumextended
LibellulidaeLabiumextended
DragonflyMercuryProject–SamplingGuide
Diagram:KenSolteszNote:TheillustrationshowsatypicalGomphid,withpaddle---shapedantennaethathavefewer segments;most larvaehave long, filamentous antennaewithmoresegments.
DragonflyMercuryProject–SamplingGuide
5. Store&Send• Inthefield,putdragonflysamplebagsintoacoolerwithicepacksorwetice,
beingcarefulnottosquishdragonflylarvae.• Uponreturntothelaboratoryoroffice,storedragonflylarvaeinafreezeruntil
shipping.• Donotopentheinnerdragonflybags!Ifyouwanttocheckyouridentification,doso
quickly,beforefreezing,andthroughtheinnerbag.Donotallowdragonflysamplestothawandrefreeze;itisimportantthattheyremainfrozenonceplacedinthefreezer.
• Dragonflysamplescanbestoreduntilallsitesaresampled,thenmustbeshippedinacooleronDRYICE.It’sbesttoplacesamplesunderthedryice,andfillanyairspaceinthecoolerwithcrumplednewspaper.Includeacopyofyourfieldsheetintheshipment,andkeepacopyforyourself.Alternatively,thedatasheetscanbescannedandemailedto the coordinating labwith theoriginals included in the shipment. Contactus if youhavetroublefindingdryice,orhavequestionsaboutit.
• Note:Whenshippingsamplesondryice,youmustcheckthedryiceboxontheFedEx®label.Additionally,thecoolermusthavetheDryIcelabel(includedinsamplingkit)affixedwithwaterprooftape.
• Contactthecoordinatorwhenashipmentisreadytoverifythatsomeoneisavailabletoaccepttheshipment.ShipmentofcoolersshouldbesentFedExovernight(withpre-paidreturnlabelincludedinkit).
CoordinatorforShipmentofSamples:JamesWillacker([email protected]);541-750-0957(office)or541-243-3606(cell)ShippingAddress:USGSForestandRangelandEcosystemScienceCenterAttn:JamesWillacker3200SWJeffersonWayCorvallis,OR97331
6. Usethedata!Weaimtomakepreliminarypark-specificdataavailable~1yearafterthesampleshavebeencollected.Workwithyourcitizenscientiststodescribethefindingsatyourpark.Submitentriestothelocalsciencefair.Feelfreetocontactusifyouwanttodiscussyourpark’sdataandhowtoincorporateitintoparkprograms.UMaineandUSGSwillinterprettheresultsforthenationalparkseachprojectyear,throughwebinarsorotheroutlets.
DragonflyMercuryProject–SamplingGuide
AppendixA.NetsParticipating parks must provide their own sampling nets. There are many options for nets, ranging fromexpensiveD-netstomodifiedinexpensivebaitwellnets.Seeafewoptionsbelow.Noneisnecessarilybetterthantheother,sochooseaccordingtosubstrate(ifknown),resourceavailability,andgroupsize.1. TheD-Net:LaMotteD-Net,
$70,item#138658atwww.benmeadows.com
2. Forsmallermountain
streams–ortofacilitategroupmanagement–aquarium-sizednetsmaybemoreusefulthanalargeD-Net.Thesenetscanbepurchasedforas #1lowas$4(seeexamplesatwww.carolina.com).Ahelpfultechniquemaybetoalloweachcitizenscientistorsmallgrouponenet.Eachgrouporindividualthensearchesadesignatedsectionofthestream/shore,bringingpotentialpositivesamplesbacktoacentrallocation(i.e.,bucket)wherethelargergroupcancollectivelyobserve,identify,andselect.
3. Agood,inexpensivenet(specifictodragonfly
larvaeandnotgoodforquantitativebiomonitoring)canbemadebymodifyingabaitwellnet–stretchthenetacrosslikeapoolskimmerandattachwithplasticziptiesorsmallnylonstrings.Thisworkswellbecauseitwon’tplugupwithsediment(drainsquickly)andretainslargeinvertebrateslikedragonflylarvae.Costis$20-30andtheyareavailableonline(http://www.forestry-suppliers.com/product_pages/View_Catalog_Page.asp?mi=5061)oratlocalfishingshops.Thesenetsarelightweightaswell.
#3#2
DragonflyMercuryProject–SamplingGuide
AppendixB.Howcleanis“clean”?Mercuryisallaroundus:inourhair,insoil,leaves,etc.Weneedtokeepeverythingcleansotonotcontaminateoursamples.Readonforrecommendationsongettinggearready....
Howweareusingtermsinthisproject:• Clean:Materialsandsuppliespreparedinsuchawaythattheywouldnothavecontaminationthatcouldaffectouranalysesandinterpretationofmercury.
• New:Weusethistermwhenreferringtoitemslikeplasticspoonsanddishpans.By“new”,wemeanthatthematerialsareeitherbrandnewfromthestore,oryouknowtheyhaveonlybeenusedtoholdnaturalwaterandinvertebrates,similartowhatisdoneinthisstudy.Forinstance,dishpansmighthavebeenusedbeforeinyourfieldsamplingformacroinvertebrates,whichisfine.Whatwemeanby“new”isthatthisisadesignatedfieldsupply,notanolddishpanorspoonthatwasusedwithfood,ormaterialsthatyoureallydon’tknowabout.
Levelofcleaningneededforeachfielditem:Manysuppliesare“disposable”:plasticspoons,gloves,andextrabags.Theycometoyoupre-cleanedandreadytotakeintothefieldtousejustastheyare.Theseshouldbemarkedas“used”whenyouaredoneatonesiteandheadingtothenext.Eachsitehasitsownsamplingkitandyoushouldstartfreshateachnewsite.Agoodpracticeistodrawan“X”onthetrashbagwithanysuppliesthathavebeenusedatasite,tokeepthemfrombeingconfusedwithfresh,un-usedsupplies.
Beforeyougointhefield:• Fornewsupplies(e.g.,ifyouboughtadishpan),rinseitoutthreetimeswithhottapwaterbeforetakingitoutinthefield.Thenrinseitinsitewateraswithitemsyouarere-using(see“New”).
• Youcanusenewgarbagebagstotransportlargeitemsintothefield(tominimizeroad/traildirt),andtostoreequipmentbetweensamplingdays.Justbesureeverythingisdryfirsttoavoidmildew.
Beforeyourfirstsiteorbetweensites:(evenwithnewitems)IfyouareheadingfromSiteAtoSiteB,youhavesomefieldgearthatwillcontactthesamplesthatneedstobere-used:nets,anddishpans.WhatdoyoudotoensureyouarenotintroducingSiteAcontaminationatSiteB?Andfornewitems,howdoyouensurethatthere’slimitedcontaminationfromthestoreorfromtransit?• First,youwillprobablyhavedisinfectedyournets,boots/waders,anddishpansafteryoufinishedworkatSiteA.Manyparksrequiredisinfectiontoreducethespreadofamphibiandiseaseorotheraquatic-bornepestsfromsitetosite.Eachparkhasitsownpolicyandrequirements.Often(butnotalways)thismeanssoakingorsprayingnets,boots/waders,anddishpanswithableach-watersolution.Checkyourpermitforprocedures.
• Then,youwillprobablytossyourwet,disinfectedgearinthebackofavehicleandmaybedrivedowndustyroadsorhikethroughthewoodstogettoSiteB.
• WhenyougettoSiteB(oryourfirstsite,fornewgear),locateanareaofthepondawayfromthattargetsamplinglocation,ordownstreamofitinthecaseofariver/stream,andrinseyourwaders.
• Havetherestofthecrewrinsetheirboots,waders,andnetsinpond/streamwaterawayfromtheprobablesamplingsiteordownstream.Dunkgearinthewaterthreetimestorinse.Thenproceedwithsampling.
Withinasite:IfyouareatSiteA,andyouarebaggingallofyourdragonflylarvaefromSiteA,youcanrinseyourplasticspooninlake/streamwaterbetweensamples.ThesameistrueforyourNitrile(purple)gloves.Butdon’tusethoseSiteAspoonsorglovesatSiteB–theyarecontaminatedwithSiteAmaterials.
DragonflyMercuryProject–SamplingGuide
AppendixC.ResearchPermitApplication–exampleRPRStext
Projecttitle:TheDragonflyMercuryProject–engagingcitizenscientistsinmonitoringmercurycontaminationinNationalParksPurpose:Mercury,initstoxicmethylatedform,isapotentneurotoxinthatisdeliveredtoecosystemsviadepositionfromaglobalatmosphericpool,andultimatelybioaccumulatesinaquaticandterrestrialfoodwebs.InthenortheasternU.S.,researchsitesin‘pristine’areashavefishandotherbiotathatexceedthresholdsconsideredsafeforhumanconsumptionorwildlifeprotection.AllNewEnglandstates,andmanyotherstates,havestatewidefishconsumptionadvisorieswithrespecttomercurybecause,inlargepart,ofthedifficultyinpredictingwhichsystemsarelikelytobemostaffected.Thisstudywillbepartofongoingcitizenscienceresearchthatisevaluatingtheutilityofdragonflylarvae(Odonata:Anisoptera)asindicatorsofmercurystatus.Ourearlydataarebeingusedtodevelophypothesesregardingwhethermercuryvarieswithodonatelarvalbodysizeorbyfamily;orwhetherasite’slandscapesettingdrivesvariabilityinmercuryinodonatelarvae.Todate,researchhasbeenprimarilycarriedoutbycitizenscientistsinnationalparksinthepilotprogram;thispermitrequestisinsupportofanefforttoexpandtheworkbeyondthepilotparkstoidentifybroader-scalespatialpatternsandbetterunderstandtheutilityofthisbio-sentinel.Summaryofproposedfieldmethodsandactivities:AquaticmacroinvertebratesaretypicallycollectedusingstandardD-netswith500micronmeshordipnetswithlargermesh(ifdragonflylarvaeonlyaretargeted),and/orbyinspectionofcobbles,submergedoremergentvegetation,andwoodymaterial.Individualdragonflylarvaearepicked(withglovesorpre-cleanedforcepsorspoons)fromnetsanddoublebaggedinzippersealbags.Individualsarefrozenuntilshipmentandanalysis.Bodylengthofindividualsandidentificationtotaxonomicfamilymaybedoneatthefieldsiteorafterreceivedinthelab,bypriorarrangement.Atthefieldsite,individualsnotselectedforanalysisareimmediatelyreturnedtothewaterbodyoforigin.RepositoryType:WillbedestroyedthroughanalysisordiscardedafteranalysisObjectsCollected:
− Odonata:anisoptera(dragonfly)nymphs;45perparkpersamplingouting.(15persite,2–3sites).
LocationInformation:
− Listsites:(Specifictoeachpark;tobedetermined.)− Access:(Specifictoeachpark;accessbyfootonroadsortrails.)
Wherewilldata,maps,photos,etc.(notspecimens)resideuponcompletionofthisproject?Dataarestoredinoriginallaboratoryfiles,ontheprojectPIscomputer(andassociatedbackups),andwillbeuploadedtoIRMAafterfinalqualityassuranceandpublicationofresults.Datacollectedbystudents/teachersfromsitesinthepilotstudiesarealreadyavailableonIRMA.
DragonflyMercuryProject–SamplingGuide
AppendixD.HowtouseiNaturalist
From hikers to hunters, birders to beach-combers, the world is filled with naturalists, and many of us record what we find. What if all those observations could be shared online? You might discover someone who finds beautiful wildflowers at your favorite birding spot, or learn about the birds you see on the way to work. If enough people recorded their observations, it would be like a living record of life on Earth that scientists and land managers could use to monitor changes in biodiversity, and that anyone could use to learn more about nature. That's the vision behind iNaturalist.org. So if you like recording your findings from the outdoors, or if you just like learning about life, join us! What Are Observations? An observation records an encounter with an individual organism at a particular time and location. This includes encounters with signs of organisms like tracks, nests, or things that just died. When you make an observation, you’ll record:
DragonflyMercuryProject–SamplingGuide
Make an observation from the Web
Start by tapping Add from the dropdown in the top menu
Type a name for what you saw and choose from one of the choices. If you can’t find what you’re looking for, leave it blank or use some placeholder text. If you entered a scientific name that's not recognized, click ‘Search external name providers’ to first import the organism from elsewhere. Let us know if the organism you observed was captive or cultivated.
Type in an address and click ‘Search’ in order to calculate coordinates and an accuracy circle
describing where you were. You'll probably also need to zoom into the map and manually adjust the marker position and accuracy circle size.
Adding evidence for what you saw in the form of photos or sounds provides. You can upload photos from your hard drive or import photos or sound from your other social networks by linking them.
Use the calendar to enter when you observed the organism.
DragonflyMercuryProject–SamplingGuide
Save your observation. How to Make an Observation with your iPhone
Tap observe.
Add one or more photos as evidence.
Enter what you saw or leave this blank.
When you saw it should be added automatically.
Where you saw it should be added automatically, if not check Privacy in the Settings app.
Save your observation.
Upoad to share with the community. This should happen automatically, if not tap the button. You can turn off autoupload from the app settings from the Me tab.
Check back for activity on your observation from the community.
DragonflyMercuryProject–SamplingGuide
Make an observation from your AndroidTap the observe button from the My Observations tab.
Add one or more photos as evidence.
Enter what you saw or leave this blank.
When you saw it should be added automatically.
Where you saw it should be added automatically, if not check Privacy in the Settings app.
Save your observation.
Sync to share with the community. This should happen automatically, if not tap the button. You can turn off autoupload from the Settings tab.
Check back for activity on your observation from the community.
DragonflyMercuryProject–SamplingGuide
AppendixE.SacrificingDragonflyLarvaeforScience:WhatistheImpact? Dragonfliesaretypicallyfoundinveryhighdensities,andourprojectsamplesarelativelytinynumberperwaterbody.Evidence:
• Rangeis20-200individuals/m2dependingontimeofyearandspecies1,2
Butwedidn’tfindmanylarvaeinoursite!• That’sOK–it’slikelythatthetimingisoffandyoumightbebetweenpopulationsthatare
emergingandthosethatarelayingneweggs.• Ifyouarelookingfor20minutesandfindnothing,tryanotherspotalongtheshore.Thereare
hotspotsinmicrohabitats.
Dragonfliesareprolificreproducers(rstrategists):theyproducealotofoffspringbecauseitisexpectedthatmostwon’tsurvivetoadulthood.Evidence:
• RearingstudiesinMaine:asinglefemalelays400-1600eggsandcandothismorethanonceperseason.
• Otherstudiesshowthatfemalescanlayfrom800-2600eggsovera9dayperiod.3• Innature,>95%oflarvaedon’tsurvivetoadulthood.4,5
Otherthreatssuchasvehicletrafficcanposeasignificantthreattodragonflies:• Toadragonfly,roadsarewarm,sunnyclearingswithampleflyinginsectprey;thus,dragonflies
oftenuseroadsasrestingandforagingsites.• InonestudyatAcadiaNationalPark,researchersfoundthatvehiclecollisionswithflyingadult
dragonflieswerecommonnearroadways:atleast3.2Odonataperkilometerofroadwerestruckeachdayduringthebreedingseason,alonghigh-trafficroadwaysnearfreshwaters.6Further,thesemortalityrateswerelikelyunderestimatesbecauseresearcherswereunabletocountinsectsthatremainedstucktovehicles,andweatherwasunusuallyrainyduringthestudy.
Theimpactofthisprojectondragonfliesisexpectedtobeminimal.• Wesamplethesmallestnumberofindividualsthatarestatisticallymeaningful.• Poweranalysesareusedtooptimizethenumberofsamples.• Otherfactorsareresponsibleforevenhigherratesofmortality(cannibalism,predation,
automobilecollisions(seeabove))• Dragonfliesareubiquitousandfoundinsomanyhabitatsaroundyourregion–evenurban
sites,irrigationditches,cattleponds,coldregions,andnutrient-enrichedsites.
1Benke,A.C.,&Benke,S.S.(1975).Comparativedynamicsandlifehistoriesofcoexistingdragonflypopulations.Ecology,56(2),302-317.2Benke,A.C.(1976).Dragonflyproductionandpreyturnover.Ecology,57(5),915-927.3Hopper,K.R.,etal.Crowley,P.H.,&Kielman,D.(1996).Densitydependence,hatchingsynchrony,andwithin-cohortcannibalisminyoungdragonflylarvae.Ecology,77(1),191-200.4Wissinger,S.A.(1988).Lifehistoryandsizestructureoflarvaldragonflypopulations.J.N.Am.Benth.Soc.,7(1),13-28.5McPeek,M.(2012).DartmouthCollege,pers.comm.6Remsburg,A.,&Blaker,P.(2009).OdonatacommunitiesofAcadiaNationalParkandMtDesertIslandfromexuviaandroadsidesurveysin2009.Unpubl.projectreport,UnityCollege,UnityME.