In-situ toxicity to woodland crayfish (Orconectes hylas)

15 Pages·2013·0.75 MB·English

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USGS Staff -- Published Research US Geological Survey
2009
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Ann L. Allert
United States Geological Survey, [email protected]
James F. Fairchild
United States Geological Survey, [email protected]
Robert J. DiStefano
Central Regional Office and Conservation Research Center
C.J. Schmitt
United States Geological Survey, [email protected]
William G. Brumbaugh
United States Geological Survey, [email protected]
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Allert, Ann L.; Fairchild, James F.; DiStefano, Robert J.; Schmitt, C.J.; Brumbaugh, William G.; and Besser,
J.M., "Ecological effects of lead mining on Ozark streams: In-situ toxicity to woodland crayfish
(Orconectes hylas)" (2009). USGS Staff -- Published Research. 531.
https://digitalcommons.unl.edu/usgsstaffpub/531
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Ann L. Allert, James F. Fairchild, Robert J. DiStefano, C.J. Schmitt, William G. Brumbaugh, and J.M. Besser
This article is available at DigitalCommons@University of Nebraska - Lincoln: https://digitalcommons.unl.edu/
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ARTICLE IN PRESS
EcotoxicologyandEnvironmentalSafety72(2009)1207–1219
ContentslistsavailableatScienceDirect
Ecotoxicology and Environmental Safety
journal homepage: www.elsevier.com/locate/ecoenv
Ecological effects of lead mining on Ozark streams: In-situ toxicity
to woodland crayfish (Orconectes hylas)
A.L. Allerta,(cid:2), J.F. Fairchilda, R.J. DiStefanob, C.J. Schmitta, W.G. Brumbaugha, J.M. Bessera
aUSGeologicalSurvey,ColumbiaEnvironmentalResearchCenter(USGS/CERC),4200NewHavenRoad,Columbia,MO65201,USA
bMissouriDepartmentofConservation,1110SouthCollegeAvenue,Columbia,MO65201,USA
a r t i c l e i n f o a b s t r a c t
Articlehistory: The Viburnum Trend mining district in southeast Missouri, USA is one of the largest producers of
Received22April2008 lead–zincoreintheworld.Previousstreamsurveysfoundevidenceofincreasedmetalexposureand
Receivedinrevisedform reducedpopulationdensitiesofcrayfishimmediatelydownstreamofminingsites.Weconductedanin-
28July2008 situ 28-d exposure to assess toxicity of mining-derived metals tothewoodland crayfish (Orconectes
Accepted28August2008
hylas). Crayfish survival and biomass were significantly lower at mining sites than at reference and
Availableonline20December2008
downstreamsites.Metalconcentrationsinwater,detritus,macroinvertebrates,fish,andcrayfishwere
Keywords: significantlyhigheratminingsites,andwerenegativelycorrelatedwithcagedcrayfishsurvival.These
Crayfish results support previous field and laboratory studies that showed mining-derived metals negatively
Orconecteshylas
affectO.hylaspopulationsinstreamsdrainingtheViburnumTrend,andthatin-situtoxicitytestingwas
Lead–zincmining
avaluabletoolforassessingtheimpactsofminingoncrayfishpopulations.
Lead
PublishedbyElsevierInc.
Zinc
Cadmium
Nickel
Cobalt
In-situtoxicity
1. Introduction Wixson,1977)showedthatmetalexposurestoaquaticbiotahad
occurred within the Viburnum Trend. Subsequent investigations
Extensive deposits of lead (Pb) ore have been mined in have documented elevated metal concentrations in water,
Missouriformorethanthreecenturies.MiningoftheViburnum stream sediments, and aquatic food chains (Besser et al., 2006;
Trend in southeast Missouri, USA, which contains economi- Brumbaugh et al., 2007; Schmitt et al., 2007a,b), and the
cally significant Pb–zinc (Zn) ores, began in the 1950s. By 1970, loss of biota including crayfish (Allert et al., 2008) and other
the Viburnum Trend was the largest Pb-producing region in macroinvertebrates(unpublisheddata;B.Poulton,USGS, Colum-
the world (Ryck and Whitley, 1974). Deposits within the bia, MO). In addition, sportfish including smallmouth bass
Viburnum Trend also contain considerable quantities of copper- (Micropterus dolomieu), longear sunfish (Lepomis megalotis), and
(Cu),cobalt-(Co),andnickel-(Ni)bearingminerals(Jessey,1981). suckers (Catostomidae) are known to contain Pb at concentra-
Although the mining district remains a major producer of Pb, tions exceeding recommended human food consumption levels
and a minor producer of Zn and Cu (Missouri Department of (MissouriDepartmentofHealthandSeniorServices,2005).
Natural Resources, 2004), there is no commercial recovery Crayfish hold an intermediate trophic position and facilitate
ofCoorNi(Kuck,2004;Shedd,2004).Weconductednoanalyses the flow of nutrients and energy in aquatic ecosystems (Lodge
of copper because previous studies have indicated that it et al., 1995). Crayfish process a significant portion of organic
is a minor component of metal contamination from mining matter in streams (Usio, 2000), breaking allocthonous and
in the Viburnum Trend (Besser et al., 2008; Brumbaugh et al., autochthonousmaterialintosmallerparticlesthatareultimately
2007). consumedbyaquaticinsects,snails,and microbial fauna(Huryn
Although modern mining practices incorporate efficient ex- andWallace,1987;Parkynetal.,2001).Crayfisharethedominant
traction technologies and operate within environmental regula- invertebratebiomassinsomeOzarkstreams(Rabenietal.,1995)
tions, early studies (Duchrow, 1983; Ryck and Whitley, 1974; and are the primary food source for several centrarchid fishes
(WhitledgeandRabeni,1997).Crayfisharealsopreyformorethan
200speciesofinsects,arachnids,amphibians,fish,reptiles,birds,
(cid:2)Correspondingauthor.Fax:[phone]. and mammals in and around North American water bodies
E-mailaddress:[email protected](A.L.Allert). (DiStefano,2005).
0147-6513/$-seefrontmatterPublishedbyElsevierInc.
doi:10.1016/j.ecoenv.2008.08.005
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1208 A.L.Allertetal./EcotoxicologyandEnvironmentalSafety72(2009)1207–1219
The southern-most mines within the Viburnum Trend are exposure through aqueous and dietary pathways, and (2) to
located in the headwaters of the Black River watershed (Fig.1). evaluate the effects of mining-derived metals on survival and
Thewoodlandcrayfish(Orconecteshylas)isendemictotheBlack growthofO.hylas.
River watershed (Pflieger, 1996) and occurs at high densities
(25–37/m2) in riffle habitats of Ozark streams (DiStefano et al.,
2002). Allert et al. (2008) reported reduced densities of O. hylas 2. Methods
(0–2/m2) at sites immediately downstream of mining activities
comparedwithdensities(15–20/m2)atsitesupstreamofmining 2.1. Studyarea
oratsitesgreaterthan10kmdownstreamofminingareasinthe
Black River watershed. These findings suggest that mining Crayfishcagesweredeployedfrom30Juneto28July2005atsevensitesin
activities are adverselyaffecting crayfish populations. We there- three tributaries of the Black River (Table 1). Previously collected physical,
chemicalandbiologicaldata(Allertetal.,2008;Besseretal.,2006;Brumbaugh
fore conducted chronic in-situ exposures of O. hylas in two
etal.,2007)wereusedtoidentifyreferencesites(sitesupstreamofknownmining
tributaries of the Black River watershed with the following
activities);miningsites(sitesimpactedbyminingactivity);anddownstreamsites,
objectives: (1) to evaluate crayfish responses relative to metal where possible biological recovery may occur. Two sites were designated as
Fig.1. MapofstudysitesintheBlackRiverwatershedofMissouri,USA.
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A.L.Allertetal./EcotoxicologyandEnvironmentalSafety72(2009)1207–1219 1209
Table1
SamplingsitesandcagelocationsinthreetributariesoftheBlackRiver.
Site Stream Latitude,longitudea Prominentminingfeatureupstream Stream Streamdistance Typeof
order fromtailingsor site
mine(km)
SC2 StrotherCreek 37136007.200,91101040.800 Effluentpond;Buicktailings 2 3.7 M
WF1 WestFork 37130039.600,91109043.200 None 2 NA R
WF3 WestFork 37129049.200,91105013.200 BrushyCreektailings;WestForktailings 2 2.2 M
WF4 WestFork 37129013.200,91103003.600 BrushyCreektailings;WestForktailings 2 10.1 D
BF1 BeeFork 37126043.400,91106029.400 None 1 NA R
BF3 BeeFork 37126028.700,91105038.000 Fletcherclarificationdamandtailings 1 0.4 M
BF5 BeeFork 37127036.000,91101030.000 Fletcherclarificationdamandtailings 2 7.4 D
Latitude,longitudeasdeterminedbyglobalpositioningsystem(GPS)receiver.R¼referencesite;M¼miningsite;D¼downstreamsite;NA¼notapplicable.
a(GPS;710m)basedontheWGS84geodeticdatum.
Fig.2. Diagramofcage.Cageframeconsistsoflow-densitypolyethylene(LDPE)strips(0.635-cmwidth)heatweldedtostainless-steel-woven-wiremesh(2.7-mmdiagonal
meshopening).Cageincludesahingeddoorwith6-mmnylonshockcordusedtosecuredoorbyfasteningaroundstainlesssteelmachinescrewsinstalledoncageframe
andcagedoor.Thefeeding/stockingportconsistsofatwist-offpolyvinylchloride(PVC)cap.Polyethylenemeshpacks(1.27-cmdiagonalmeshopening)filledwithrolledup
scourpads(forcrayfishrefuge),organicmaterial,andcobblewereinsertedintocagepriortostockingwithcrayfishandsecuredtobottomofcagewithstainless-steelwire.
reference sites (WF1, BF1); three as mining sites (SC2, WF3, BF3); and two as 2.3. Toxicitytest
downstream sites (WF4, BF5). Sites classified as mining sites were 0.4–3.7km
downstreamofminingactivity,whereasthoseclassifiedasdownstreamsiteswere
atleast7kmdownstreamofmining-relatedactivity. A28-din-situtoxicitytestwasconductedwithjuvenileO.hylas,atsevensites
intheWestFork,BeeFork,andStrotherCreekoftheBlackRiverwatershed(Fig.1).
Crayfishwereexposedinhemicylindrical(0.28-m2)cagesconstructedofstainless-
2.2. Crayfishcollectionandrearing steelwiremesh(2.7-mmdiagonalopening)andpolyethylene(LDPE)reinforcing
strips(Fig.2).Wecollectedcobblesubstrates(2.5–7.5cm)andapproximately10-g
Ovigerous O. hylas females were collected from the reference site on the organicmaterial (henceforthdetritus)fromeachsitetoprovidebothfoodand
West Fork (WF1) in early April in 2005 and returned to the US Geological shelterforcagedcrayfish.Cobble,detritus,andthreepolyethylenescourpadswere
Survey’s Columbia Environmental Research Center (CERC) located in Columbia, placedinthreepolyethylene-meshpacks(15-cmlength(cid:2)30-cmwidth;1.27-cm
MO, USA. Sixteen females were held in individual 2-L flow-through aquaria diagonalopening),whichwereclosedusingplasticcabletiesandsecuredtothe
withCERCwellwater(temperature181C,pH7.7,alkalinity254mg/LasCaCO3, bottomofeachcageusingstainless-steelwire.Priortoplacingorganicmaterial
hardness 286mg/L as CaCO3) and fed frozen brine shrimp (San Francisco Bay intothemeshpacks,allpredatoryinsects(i.e.,OdonataandPlecopteralarvae)
Brand,Inc.,SanFrancisco,CA,USA)adlibitumdaily.Uponhatchinganddetaching were removed. The bottom of the cages were buried 2–4-cm into the stream
from the adult females, juvenile crayfish were placed in a fiberglass tank sedimenttoexposecrayfishtosub-surfacewaterandtoanchorthecages.Minced
filled with CERC wellwater and fed flake food (Ziegler Brothers, Inc., Gardner, fish(largescalestonerollers,Campostomaoligolepis;henceforthstonerollers)from
PA,USA)adlibitumdailyuntiltheirbodywidthwasgreaterthan2mm.Priorto eachsitewereaddedtoeachcageweeklyinincreasingincrementstomaintain
stockingcrayfishintocages,asubsetofcrayfishavailableforstockingintocages dietary rations proportional to anticipated crayfish biomass (0.1, 0.1, 0.2, 0.4g
(n¼88) was measured for mean carapace length (CL, from tip of rostrum to mincedfishinweeks1,2,3,and4,respectively).Tenjuvenilecrayfishwereplaced
posterioredgeofthecephalothorax,tothenearest0.1mm)andweighed(tothe in each of the six cages at each site except at SC2, where seven cages were
nearest0.01g). deployed. Three cages (n¼30 crayfish) were sampled on both day14 and 28,
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exceptatSC2,wherefourcagesweresampledonday28.Crayfishweremeasured macroinvertebrates,stonerollers,andcrayfisharelistedinTableS-2.Allmeasured
(CL and wet weight) and frozen for metal analyses. Test endpoints included concentrationsexceededtheMLDs.
survival and growth. Biomass (i.e., standing crop) at day 28 was estimated Recoveriesoftheelementsfromreferencematerials(fish,mussel,oyster,plant,
bymultiplyingthenumberofsurvivorsbythemeanwetweightofsurvivorsat andplankton)rangedfrom87%to115%.Relativepercentdifferences(RPDs)for
eachsite. replicateanalyseswereo26%forallelementsexceptforPbanalysesofonesample
of detritus (50%) and one sample of macroinvertebrates (85%). Instrumental
precision, estimated by determining the RPDs from the duplicate analysis of
2.4. Wildcrayfishcollections detritusandbiotadigestates,waso4%.Recoveriesofmethodspikesforallfive
metalsin14separatespikedsamplesofallthesampletypesanalyzedaveraged
Wildcrayfish(O.hylas)werecollectedondays0and28ofthetoxicitytestat 97%.Post-digestionoranalysisspikerecoveriesrangedfrom83%to104%.Asa
eachsitetocompareCL,wetweight,andmetalconcentrationsofwildcrayfishto checkforpotentialinterferences,dilutionpercentdifferences(DPDs)basedon5X
thosestockedincages.Crayfishwerecollectedbydisturbingsubstratedirectly dilutionsof detritus and biotasample digestateswere determined;DPDs were
upstreamofasmallkickseine(1-mlength(cid:2)1.5-mheight)with3-mmdeltamesh o10%forallmetals.Blank-equivalentconcentrations(BECs)fordigestionblanks
(FlindersandMagoulick,2005).Wildcrayfishwerecollectedwithin200mofcage werelessthancorrespondingMLDs;thereforesampleresultswerenotcorrected
placement.Collectionsweremadeupstreamordownstreamofcageplacement, for BECs. Overall, quality assurance results indicated that the methods used
dependingonthenearestavailableriffle,andnumberofkickseinesrequiredto providedacceptableaccuracyandprecision,thusnoneofthesampleresultswere
collect30individuals.Allcrayfishcollectedwereidentifiedtospecies,measured corrected.
(CLandwetweight)andfrozenformetalanalyses.
2.7. Statisticalanalysis
2.5. Watersamples
StatisticalanalyseswereconductedusingStatisticalAnalysisSystem(SAS)for
Surface water quality parameters (temperature, pH, conductivity, dissolved Windows(Release9.1;SASInstitute,Cary,NC,USA).Censoredvalues(oMLD)
oxygen,andturbidity)weremeasuredin-situondays0,7,14,21,and28usinga were replaced with 50%of the MLD for statistical computations and graphing.
Hydrolabs Quanta meter (Loveland, CO, USA). A sub-surface grab sample was Survivalandbiomassdataofcagedcrayfishonday28ofthetoxicitytestandthe
collected in a pre-cleaned 4-L carboy at each site on days 0, 14, and 28 for overallmeans(e.g.,datafromdays0,7,14,21,and28)forwaterquality,nutrients,
additionalwaterqualityandnutrientanalysesatCERC.Alkalinityandhardness andmetalconcentrationswereusedinthestatisticalanalyses.Nowildcrayfish
weremeasuredbytitration(APHA,2005).Sulfatewasmeasuredbycolorimetric werecollectedwithin200mofourcagesatSC2,sooursearchwasexpandeduntil
detectionwithaHachs2100spectrophotometer(Loveland,CO,USA).Recoveryof oneadultwildcrayfishwascollectedatadistancegreaterthan500mawayfrom
referencestandardsusedaslaboratorycontrolsamplesforsurfacewaterquality thecages;thereforeitwasnotusedinthestatisticalanalysesofCLandwetweight
parameters (pH, conductivity, dissolved oxygen, turbidity, alkalinity, hardness, ofwildcrayfish.However,datafromthisindividualispresentintablesandfigures
andsulfate)rangedfrom90%to118%.Overall,detectionandrecoveriesofwater forcomparativepurposes.Alldataweretestedfornormalityandhomogeneityof
qualityparameterswerewithinacceptablecriteria,thusnoneofthesampleresults variance using the PROC UNIVARIATE module in SAS. Data were not normally
werecorrected. distributed, therefore rank transformation were used in statistical analyses.
Samples for dissolved nutrient analyses were filtered through 0.4-mm Differences in caged crayfish survival and biomass among sites and groups of
polycarbonatefiltersundervacuumpressurewithin4daysofcollection.Nutrients sitesweretestedusingnestedanalysis-of-variance(ANOVA;cagesnestedwithin
were measured in surface water samples with a Technicons Autoanalyzer site), with site considered a fixed effect. Differences in caged crayfish survival
(Tarrytown,NY,USA)usingcolorimetricdetection(APHA,2005).Totalammonia among groups of sites were tested as planned non-orthogonal contrasts using
(NH3) was analyzed using a salicylate/nitroprusside colorimetric reaction. singledegree-of-freedomF-tests.Themeansquareforcagesurvivalwithinsite
Dissolvednitrite/nitrate(NO2/NO3)wasmeasuredfollowingcadmiumreduction wasusedinalltests,whichwereconductedusingthePROCGLMmoduleinSAS.
and measured using colorimetric reaction (APHA, 2005). Soluble reactive Differences in crayfish survival and biomass among individual sites were also
phosphorus (SRP) was determined using the automated ascorbic acid method evaluatedwithDuncan’smultiplerangetest.Differencesinwildcrayfishgrowth,
(APHA,2005).Samplesfortotalphosphorous(TP)andtotalnitrogen(TN)were waterquality,nutrient,andmetalconcentrationsamonggroupsofsiteswerealso
digestedinsodiumhydroxideandpotassiumpersulfatethenanalyzedusingthe testedusingthesameprocedures.Differencesinmortality,sizeofcrayfish,water
automated ascorbic acid and the automated cadmium reduction methods, quality, nutrient, and metal concentrations were tested on a stream-by-stream
respectively(APHA,2005).Dissolvedorganiccarbon(DOC)wasanalyzedusinga basisusingtheKruskal–Wallistest.Resultsweresimilartothosethatincludedall
persulfate/UVdigestionfollowedbycolorimetricanalysisofCO2.Methodlimitsof seven sites; therefore we list only exceptional results in Table S-3. Finally,
detectionlimits(MLDs)fornutrientsarelistedinTableS-1.Recoveryofreference associationsamongcagedcrayfishsurvivalandbiomassonday28,wildcrayfish
standardsusedaslaboratorycontrolsamplesfornutrientsrangedfrom100%to (excludingSC2)CLandwetweight,waterquality,nutrient,andmetalconcentra-
128%,exceptforonestandardforNH3(1mgN/L)thatwas145%.Overall,detection tionswereexaminedwithSpearman’scorrelationanalysis.Asignificancelevelof
and recoveries of nutrients were within acceptable criteria and all measured Po0.05wasusedtojudgeallstatisticaltests.
concentrationsexceededMLDs,thusnoneofthesampleresultswerecorrected
forrecovery.
3. Results
2.6. Metalconcentrations
3.1. Toxicitytest
Water samples were filtered for metal analyses using a polypropylene
syringeandfiltercartridge(0.45-mmporesize)intoapre-cleanedpolyethylene Meanpercentsurvivalofcagedcrayfishonday28(henceforth
bottle on-site, and placed on ice. Water samples were subsequently acidified survival)wassignificantlygreateratreferencesites(90%)thanat
to1%(v/v)withnitricacid(J.T.BakerInc.,Phillipsburg,NJ,USA)within4days
mining sites (39%; Table 2; Fig. 3), and decreased sharply with
ofcollection.
SurfacewatersampleswereanalyzedforPb,Zn,Cd,Ni,andCobyinductively- increasing metal concentrations in surface water (Fig. 4). When
coupled plasma-mass spectrometry (ICP-MS) (Brumbaugh et al., 2007; May analyzedonastream-by-streambasis,survivalattheminingsite
etal.,1997).Calibrationverification,methodlimitsofdetection,andrecoveries (BF3) in the Bee Fork was significantly lower; however, survival
ofmetalsin referencesolutions,duplicates,andspikeswereallwithinaccept-
wasnotsignificantlylowerattheminingsite(WF3)intheWest
able criteria and all measured concentrations exceeded MLDs (Brumbaugh
Fork(TableS-3).
etal.,2007).
Detritus, macroinvertebrates (i.e., Ephemeroptera, Odonata, Plecoptera, MeanCLandwetweightofcagedcrayfishatallsitesonday28
Megaloptera,Trichoptera,Diptera,andChirononmidae),stonerollers,andwhole were significantly greater than those on day 0 (n¼88;
crayfishfromeachsitewereanalyzedforPb,Zn,Cd,Ni,andCobyICP-MS(Besser CL¼6.770.11; wet wt.¼0.0670.003), except meanwet weight
etal.,2006;Brumbaughetal.,2005).Tissueswerelyophilizedandreducedtoa
ofcrayfishatSC2(Table2).MeanCLandwetweightofcrayfishat
coarsepowderbymechanicalcrushinginaglassvialwithaglassrod.Neither
exoskeletonsnorgutcontentsofanyofthebiotawereremovedbeforeanalysis.A day 28 were significantly greater at downstream sites than
drymassof0.25gfromeachcompositedsamplewasdigestedusingconcentrated at reference or mining sites; however, CL and weight were
nitricacidandmicrowaveheating.Qualitycontrolmeasuresincorporatedatthe not significantly different among mining and reference sites
digestionstageincludeddigestionblanks,certifiedreferencematerials,replicates,
(Table2).Meanbiomassofcrayfishatreferenceanddownstream
and spikes. A calibration blank and an independent calibration verification
sites on day 28 was significantly greater than at mining sites
standardwereanalyzedwithevery10samplestoconfirmthecalibrationstatus
oftheICP-MSduringinstrumentalanalysesofdigestates.TheMLDsfordetritus, (Table2;Fig.3).
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Table2
Number(n)oflivingcrayfish,percent(%)survival,carapacelength(CL),andwetweight(meanswithstandarderrorinparenthesis)ofcagedOrconecteshylas.
Day/site n %Survival CL(mm) Wetwt.(g) Biomass(g/m2)
Day14
Referencesites
WF1a 24 83(3)b 9.3(0.19)b 0.17(0.01)c 4.3(0.7)ab
BF1 28 93(7)a 8.5(0.22)c 0.13(0.01)d 5.3(0.8)ab
Groupmean 88(4)A 8.9(0.16)C 0.15(0.01)B 4.7(0.5)A
Miningsites
SC2 23 84(9)b 8.4(0.24)c 0.12(0.01)d 3.3(0.4)b
WF3b 27 90(6)a 9.7(0.21)b 0.20(0.01)bc 5.3(0.6)ab
BF3 17 57(23)b 9.8(0.31)b 0.18(0.02)c 4.0(1.8)ab
Groupmean 77(27)A 9.3(0.16)A 0.17(0.01)B 4.3(0.6)B
Downstreamsites
WF4 28 84(3)b 11.2(0.17)a 0.32(0.02)a 8.3(1.8)a
BF5 29 97(3)a 9.8(0.23)b 0.23(0.02)b 6.0(1.8)ab
Groupmean 92(8)A 10.5(0.01)B 0.27(0.01)A 7.3(1.3)A
Day28
Referencesites
WF1 27 90(6)a 10.5(0.33)bc 0.40(0.02)cd 1.2(3.3)c
BF1 26 90(0)a 10.8(0.37)b 0.29(0.03)de 14.7(2.3)bc
Groupmean 90(3)A 10.7(0.15)B 0.33(0.14)B 13.3(1.9)B
Miningsites
SC2c 3 7(3)d 9.2(0.03)c 0.17(0.03)e 0.3(0.2)d
WF3 20 67(9)bc 11.3(0.47)b 0.37(0.05)cd 7.7(2.5)cd
BF3 16 53(3)c 13.2(0.46)a 0.58(0.05)ab 7.7(3.5)cd
Groupmean 39(9)B 11.9(0.36)B 0.44(0.04)B 4.7(1.6)C
Downstreamsites
WF4 26 87(3)ab 14.0(0.31)a 0.71(0.05)a 32.0(4.9)a
BF5 25 77(13)ab 12.9(0.3)a 0.53(0.04)bc 22.7(5.2)ab
Groupmean 82(16)A 13.4(1.6)A 0.61(0.03)A 27.3(3.8)A
Siteswiththesamelowercaseletterandgroupsofsiteswiththesamecapitalletterarenotsignificantlydifferentforeachtestday(Po0.05).Initialstockingnumber¼30,
unlessotherwisenoted.
aStockingnumber¼29.
bStockingnumber¼31.
cStockingnumber¼40.
3.2. Wildcrayfishcollections therewerenosignificantdifferencesinTPconcentrationsamong
reference, mining and downstream sites (Table 5). Dissolved
Veryfewwildcrayfishwerecollectedatanyoftheminingsites organiccarbonwassignificantlyloweratdownstreamsitesthan
oneitherday0or28,despitesignificantsamplingeffort(i.e.,415 referenceorminingsites(Table5).
kickseines).Meanwetweightofwildcrayfishondays0and28 Analyses on a stream-by-stream basis produced slightly
weresignificantlygreateratreferenceanddownstreamsitesthan differentresultsforseveralparameters.Therewerenosignificant
mining sites; however, there was no significant difference in differencesindissolvedoxygenorDOCatsitesintheWestFork;
CLamonggroupsofsitesonday28(Table3). however, there were at sites in the Bee Fork (Table S-3). There
weresignificantdifferencesinalkalinityatsitesintheWestFork;
however, alkalinity was not significantly different at sites in the
3.3. Waterquality
Bee Fork. There was no significant difference in SRP or NH in
3
either the Bee Fork or West Fork, despite an overall significant
Mean conductivity, hardness, and sulfate concentrations in
differencewhenSC2wasincludedintheanalyses.
surface water were significantly higher at mining and down-
streamsitesthanreferencesites(Table4);whereasalkalinitywas
significantly lower at downstream sites than at reference and 3.4. Metalconcentrations
mining sites (Table 4). Reference sites had significantly higher
turbidity(Table4);however,valueswerestillverylow.Dissolved Concentrations of metals in surface water were all below
oxygen and pH at reference sites were significantly lower than surface water chronic criteria (WQC; USEPA, 2002; Table 6).
mininganddownstreamsites(Table4);however,atallthesites, Cobalt concentrations in surface water were also all below a
therangeindissolvedoxygenandpHreadingswerenarrow. proposed Canadian guideline for chronic exposure of 4mgCo/L
Ammonia, NO /NO , and TN concentrations in surface water (Nagpal,2004). However, concentrations of Pb, Zn, Ni, and Co in
2 3
from mining sites were significantly higher than reference and surfacewatersweresignificantlyhigheratminingsitescompared
downstream sites (Table 5). Soluble reactive phosphorous con- withreferenceordownstreamsites(Table6).
centrationsinsurfacewatersfrommininganddownstreamsites Concentrations of all metals in detritus, macroinvertebrates,
were significantly lower than reference sites (Table 5); however, stonerollers, and caged crayfish were significantly higher at
ARTICLE IN PRESS
1212 A.L.Allertetal./EcotoxicologyandEnvironmentalSafety72(2009)1207–1219
100
A A A A B A Lead
100 80
al 60
80 v
vi
ur 40
S
al %
v 60
vi 20
ur
S
% 40 00.01 0.1 1 10
Surface water Pb (µg/L)
20 100
Zinc
80
0
al 60
40 viv
Reference B C A Sur 40
Mining %
Downstream 20
30
2) 0
m 1 10 100
s (g/ 20 A B A Surface water Zn (µg/L)
s
ma 100
o Cadmium
Bi SC2
80
10 WF1
al 60 WF3
v
urvi 40 WF4
S
0 % BF1
Day 14 Day 28 20 BF3
BF5
Fig.3. (a)Meanpercent(%)survivaland(b)meanbiomassofcagedOrconectes 0
hylaswithingroupsofsitesondays14and28.Groupsofsiteswiththesameletter 0.01 0.1
arenotsignificantlydifferent(Po0.05). Surface water Cd (µg/L)
mining sites than at reference or downstream sites, with the 100
Nickel
exceptionofCdconcentrationsinmacroinvertebrates,andstone-
80
rollers(Table7;Fig.5).MeanconcentrationofPbinwildcrayfish
was significantly higher at mining sites than at reference or al 60
downstreamsites(Table7).MeanconcentrationsofZnandCoin vi
vi
allsampletypeswerehighestatSC2.LeadandNiconcentrations ur 40
S
werehighestatSC2and/orBF3.Metalconcentrationsinsamples %
at all mining sites were generally two to ten-fold higher in all 20
sample types compared with those at downstream or reference
0
sites.ConcentrationsofPb,Zn,andNiwerehighestindetritusand
0.01 0.1 1 10 100
were generally higher in macroinvertebrates compared with
Surface water Ni (µg/L)
stonerollers or crayfish. Metal concentrations in the West Fork
weregenerallylowerthanthoseintheBeeFork.Concentrationsof 100
Pb and Cd in fish; Zn in invertebrates, and Zn in surface water Cobalt
werenotsignificantlydifferentamongthethreegroupsofsitesin 80
theWestFork(TableS-3). al
v 60
Mean concentrations of metals in crayfish at day 0 were: Pb vi
((06..1067700.4.2mmgg/g/gddrryy wweeiigghhtt)),, NZni ((48.017700..73mmgg//gg ddrryywweeigighhtt)),,aCndd % Sur 40
Co(2.070.4mg/gdryweight).Cagedcrayfishrapidlyaccumulated 20
all metals at all mining sites (Fig. 6). Metal concentrations in
caged crayfish at SC2 continued to increase throughout the 0
exposure;however,theydidnotatWF3orBF3.Metalconcentra- 0.01 0.1 1
tions in caged crayfish on day 28 at reference and downstream Surface water Co (µg/L)
sitesweregenerallylowerthanconcentrationsincrayfishstocked
Fig.4. Relationshipbetweenmeanpercent(%)survivalofcagedOrconecteshylas
intocagesonday0.Concentrationsofmetalsincagedcrayfishat
on day 28 and mean metal concentrations in surface water at sampling sites:
day 28 were comparable to those in wild crayfish at all sites (a)lead;(b)zinc;(c)cadmium;(d)nickel;and(e)cobalt.Dashedlinesaremean
exceptatSC2,wheremetalconcentrationsincagedcrayfishwere chronicwaterqualitycriteriaforthesamplesites.
greaterthanthesinglelargewildcrayfishcollectedatSC2.
ARTICLE IN PRESS
A.L.Allertetal./EcotoxicologyandEnvironmentalSafety72(2009)1207–1219 1213
3.5. Correlationanalyses were negatively correlated with SRP, and CL was negatively
correlatedwithTP(TableS-5).
Carapace length and wet weight of caged crayfish were Meanpercentsurvivalofcagedcrayfishwasnegativelycorre-
negatively correlated with Zn concentrations in stonerollers lated with most metal concentrations in surface water, detritus,
(Table S-4). Carapace length and wet weight of caged crayfish macroinvertebrates, stonerollers, and caged crayfish (Table 8).
Biomass of caged crayfish was negatively correlated with Zn
concentrations in stonerollers and Co concentrations in caged
Table3
Number(n),carapacelength(CL),andwetweight(meanswithstandarderrorin crayfish.ConcentrationsofPb,Zn,Ni,andCoincagedcrayfishand
parenthesis)ofwildOrconecteshylascollectedatsamplingsitesondays0and28 allenvironmentalmatrixeswerehighlyinter-correlated(Table8).
ofthein-situexposure. Lead concentrations in caged crayfish were positively correlated
withPb,Zn,andNiconcentrationsinwildcrayfish(Table8).Zinc
Day/site n CL(mm) Wetwt.(g)
concentrations in caged crayfish were positively correlated with
Day0 Coconcentrationsinwildcrayfish.Survivalofcagedcrayfishwas
Referencesites negatively correlated with NH , NO /NO , TN, conductivity,
3 2 3
WF1 30 10.1(0.3)a 0.26(0.03)a hardness, and sulfate (Table S-5). Biomass of caged crayfish was
BF1 35 9.5(0.2)a 0.21(0.02)a
negativelycorrelatedwithDOC(TableS-5).
Groupmean 9.8(0.3)A 0.23(0.02)A
WetweightofwildcrayfishwascorrelatedwithCoconcentra-
tionsinwildcrayfish(TableS-4);however,CLandwetweightof
Miningsites
wild crayfish were not significantly correlated with any water
SC2 0 – –
WF3 1 5.4(–)a 0.03(–)a quality parameter or nutrient concentration (Table S-5). There
BF3 32 9.2(0.3)a 0.18(0.01)a werefewersignificantcorrelationsbetweenmetalconcentrations
Groupmean 7.3(1.9)B 0.11(0.08)B in wild crayfish and the other samples collected at the sites;
however, concentrations of Pb in wild crayfish were signifi-
Downstreamsites cantlycorrelated with most metals in all sample types analyzed
WF4 30 9.4(0.3)a 0.22(0.03)a (Table 8). Zinc concentrations inwild crayfish were significantly
BF5 43 7.1(0.2)a 0.08(0.01)a correlatedwithPb,Zn,andCdconcentrationsindetritus,andwith
8.2(0.1)A 0.15(0.07)A
Znconcentrationsinmacroinvertebrates.Nickelconcentrationsin
wild crayfish were correlated with Zn concentrations in macro-
Day28 invertebrates,andPb,Zn,andCdconcentrationsindetritus.
Referencesites
Ammonia,NO /NO ,TN,conductivity,hardness,sulfate,andNi
WF1 30 12.9(0.3)a 0.58(0.04)a 2 3
BF1 37 12.0(0.4)a 0.47(0.04)a concentrations in surface water were all significantly correlated
Groupmean 12.5(0.4)A 0.53(0.05)A (Table S-6). Soluble reactive phosphorous (SRP) was negatively
correlated with dissolved oxygen; dissolved organic carbon was
Miningsites negatively correlated with pH; and temperature was negatively
SC2 1 35.0(–) 12.1(–) correlatedwithCoconcentrationsinsurfacewater.Hardnesswas
WF3 12 12.2(0.4)a 0.48(0.05)a alsosignificantlycorrelatedwithPb,Ni,andCoconcentrationsin
BF3 2 12.3(0.2)b 0.34(0.05)a surface water. Lead concentrations in surface water were
Groupmean 12.2(0.02)A 0.41(0.07)B
significantly correlated with Co concentrations in surface water,
aswereNiandCoconcentrationsinsurfacewater.
Downstreamsites
WF4 18 13.2(0.3)a 0.56(0.04)a
BF5 30 12.3(0.3)a 0.53(0.05)a 4. Discussion
Groupmean 12.7(0.5)A 0.55(0.02)A
Siteswiththesamelowercaseletterandgroupsofsiteswiththesamecapital We documented decrease survival of caged crayfish at sites
letterarenotsignificantlydifferentforeachtestday(Po0.05). directly downstream (0.4–3.7km) of mining sites. Survival and
Table4
Waterquality(meanswithstandarderrorinparenthesis)ofsurfacewateratsamplingsites.
Site Temp(1C) PH(SU) Cond(ms/cm) DO(mg/L) Alk(mg/LasCaCO3) Hard(mg/LasCaCO3) Turb(NTU) Sulfate(mg/L)
Referencesites
WF1 23.6(0.6)a 8.04(0.03)ab 344(5)e 7.8(0.5)a 182(3)a 183(3)d 0.60(0.04)a 0.3(0.2)e
BF1 23.1(1.4)b 7.84(0.09)c 282(6)f 6.3(0.4)b 143(3)c 145(2)e 0.50(0.04)a 1(0.3)e
Groupmean 23.4(0.7)A 7.94(0.06)B 313(11)C 7.1(0.4)B 162(8)A 164(7)C 0.55(0.03)B 0.6(0.2)C
Miningsites
SC2 25.8(0.5)a 7.94(0.04)bc 858(31)a 8.0(0.2)a 117(3)d 409(11)a 0.50(0.04)a 304(10)a
WF3 24.6(0.9)a 8.09(0.03)a 418(7)cd 8.4(0.3)a 170(3)b 212(3)c 0.40(0.05)a 53(3)d
BF3 24.2(0.9)a 8.01(0.05)ab 592(15)b 8.4(0.1)a 143(2)c 247(5)b 0.40(0.06)a 129(6)b
Groupmean 24.9(0.5)A 8.01(0.03)A 623(42)A 8.3(0.1)A 143(7)A 289(26)A 0.45(0.03)A 162(32)A
Downstreamsites
WF4 24.3(0.4)a 8.10(0.04)a 395(2)d 8.3(0.1)a 165(2)b 198(1)c 0.50(0.12)a 42(1)d
BF5 22.9(0.8)a 8.05(0.04)ab 446(4)c 8.2(0.1)a 141(1)c 199(1)c 0.40(0.02)a 80(1)c
Groupmean 23.6(0.5)A 8.08(0.03)A 420(9)B 8.3(0.1)A 152(5)B 199(1)B 0.44(0.05)A 64(8)B
Temp¼temperature;Cond¼conductivity;DO¼dissolvedoxygen;Alk¼alkalinity;Hard¼hardness;Turb¼turbidity.Siteswiththesamelowercaseletterandgroups
ofsiteswiththesamecapitalletterarenotsignificantlydifferent(Po0.05).
ARTICLE IN PRESS
1214 A.L.Allertetal./EcotoxicologyandEnvironmentalSafety72(2009)1207–1219
Table5
Nutrientconcentrations(meanswithstandarderrorinparenthesis)ofsurfacewatersatsamplingsites.
Site NH3(mgN/L) SRP(mgP/L) NO2/NO3(mgN/L) DOC(mgC/L) TN(mgN/L) TP(mgP/L)
Referencesites
WF1 0.01(0.002)b 0.5(0.3)a 0.04(0.01)d 0.59(0.04)bc 0.10(0.01)c 3.7(0.6)ab
BF1 0.01(0.002)b 0.8(0.4)a 0.07(0.01)d 0.90(0.07)a 0.20(0.01)c 2.4(0.5)ab
Groupmean 0.01(0.002)B 0.6(0.3)A 0.06(0.01)C 0.75(0.06)A 0.12(0.01)B 2.9(0.4)A
Miningsites
SC2 0.06(0.01)a 0.3(0.2)a 1.2(0.09)a 0.91(0.06)a 1.30(0.08)a 4.4(0.4)a
WF3 0.01(0.002)b 0.2(0.1)a 0.13(0.01)cd 0.60(0.02)bc 0.20(0.02)c 2.8(0.8)b
BF3 0.01(0.003)b 0c 0.51(0.02)b 0.69(0.06)b 0.50(0.03)b 2.0(0.6)a
Groupmean 0.03(0.01)A 0.1(0.1)B 0.61(0.10)A 0.73(0.04)A 0.68(0.10)A 3.1(0.4)A
Downstreamsites
WF4 0.01(0.003)b 0a 0.08(0.01)d 0.57(0.03)c 0.10(0.01)c 2.0(0.2)b
BF5 0.01(0.003)b 0.2(0.2)a 0.19(0.01)c 0.50(0.04)c 0.20(0.01)c 2.6(1.2)ab
Groupmean 0.01(0.001)B 0.1(0.1)B 0.14(0.02)B 0.53(0.03)B 0.17(0.02)B 3.1(0.4)A
NH3¼ammonia;SRP¼solublereactivephosphorus;NO2/NO3¼nitrite/nitrate;DOC¼dissolvedorganiccarbon;TN¼totalnitrogen;TP¼totalphosphorous.Siteswith
thesamelowercaseletterandgroupsofsiteswiththesamecapitalletterarenotsignificantlydifferent(Po0.05).
Table6
Concentrations(mg/L;meanswithstandarderrorinparenthesis)ofmetalsmeasuredinsurfacewaters(WC)atsamplingsites,andsitehardness-adjustedchronicwater
qualitycriteria(WQC).
Site Lead Zinc Cadmium Nickel Cobalt
WC WQC WC WQC WC WQC WC WQC WC
Referencesites
WF1 0.03(0)c 4.9 4.1(2)b 197 0.03(0)b 0.37 0.13(0)c 46 0.09(0.01)de
BF1 0.03(0)c 3.8 2.5(0)b 162 0.03(0)b 0.32 0.13(0)c 38 0.14(0.01)d
Groupmean 0.03(0)B 4.4 3.3(1)B 180 0.03(0)A 0.35 0.13(0)B 42 0.12(0.01)B
Miningsites
SC2 0.71(0.02)b 11.3 56(5)a 390 0.06(0.02)a 0.65 26(2)a 90 0.64(0.05)a
WF3 0.12(0.01)c 5.7 2.5(0)b 223 0.03(0)b 0.41 1.9(0.01)c 52 0.39(0.02)b
BF3 1.59(0.06)a 6.8 6.6(0.2)b 254 0.03(0)b 0.46 6.0(0.01)b 59 0.28(0.01)c
Groupmean 0.86(0.18)A 7.9 20(7)A 289 0.03(0.01)A 0.51 11(3)A 67 0.43(0.05)A
Downstreamsites
WF4 0.04(0.01)c 5.3 2.5(0)b 211 0.03(0)b 0.40 0.40(0)c 49 0.15(0.01)d
BF5 0.03(0)c 5.3 3.9(1)b 211 0.03(0)b 0.40 0.50(0)c 49 0.12(0.01)d
Groupmean 0.04(0.01)B 5.3 3.3(1)B 211 0.03(0)A 0.40 0.46(0.02)B 49 0.13(0.01)B
Siteswiththesamelowercaseletterandgroupsofsiteswiththesamecapitalletterarenotsignificantlydifferent(Po0.05).
biomassofcagedcrayfishweresignificantlyloweratminingsites that absence of crayfishpopulations below mining siteswas the
than reference ordownstream sites, and survival was negatively resultofmetalexposureasopposedtohabitatlossduetophysical
correlated with metal concentrations in surface water, detritus, impairmentbyminewaste(e.g.,sedimentationbyminetailings).
macroinvertebrates,stonerollers,andwholecrayfish.Survivalwas Although no toxicity studies have been conducted with
alsonegativelycorrelatedwithseveralwaterqualityparameters. juvenileO.hylasinsingle-metalexposures,previousstudieshave
Our study supports previous results which found reduced shown several species of crayfish to be relatively sensitive to
densities of O. hylas populations in riffle habitats, and elevated metals, and that juvenile crayfish are more sensitive than adult
metalsconcentrations(Pb,Zn,Cd,Ni,andCo)inO.hylasatsites crayfish. Wigginton and Birge (2007) calculated 96-h median
directly downstream of mining in the Black River watershed lethal concentrations (LC ) of Cd to six species of crayfish
50
(Allertetal.,2008).Metalconcentrationsindetritusandbiotaat (Cambaridae)withameanLC of1510mgCd/Lforadultcrayfish
50
miningsitesinthisstudywerecomparabletothoseofBesseretal. and of 111mgCd/L for juvenile crayfish at a water hardness of
(2006), and decreased with distance from mining sites. Alikhan approximately 45mg/L as CaCO . Other studies (Lindhjem and
3
et al. (1990) also reported an inverse relationship between Bennet-Chambers, 2002; Mirenda, 1986a,b; Naqvi and Howell,
distancefromacontaminantsourceandmetalconcentrationsin 1993) have also reported lethal concentrations that are greater
crayfish. Conductivity, hardness, sulfate, and nitrogen ions often than the metal concentrations measured in surface waters from
are elevated below mining sites (Gray,1998; Tiwary, 2001), and ourstudystreams;however,theselaboratorystudieswereshort
maybeusefulinindicatingtheextentofminingimpacts.Finally, in duration and conducted with different species and sizes of
in-situtestingofcrayfishwasanimportanttoolfordemonstrating crayfish.Thorpetal.(1979)reportedthatlong-termexposureto5