Copper and zinc content in wild game shot with lead or non-lead ammunition

16 Pages·2017·1.07 MB·English

RESEARCHARTICLE
Copper and zinc content in wild game shot
with lead or non-lead ammunition –
implications for consumer health protection
DanielaSchlichting*,ChristineSommerfeld,ChristineMu¨ller-Graf,ThomasSelhorst,
MatthiasGreiner,AntjeGerofke,EllenUlbig,CarlGremse,MarkusSpolders,
HelmutSchafft,MonikaLahrssen-Wiederholt
GermanFederalInstituteforRiskAssessment,Berlin,Germany
a1111111111
*[email protected]
a1111111111
a1111111111
a1111111111
Abstract
a1111111111
Theaimofthisstudywastoexaminethecontaminationofgamemeatwithcopperandzinc
andestablishwhethertheuseofalternative(non-lead)ammunitioncanleadtohigheror
unsafelevelsofcopperandzincinthemeatofroedeer,wildboarandreddeer.There-
OPENACCESS
searchproject“Safetyofgamemeatobtainedthroughhunting”(LEMISI)wasconductedin
Citation:SchlichtingD,SommerfeldC,Mu¨ller-Graf Germanywiththepurposeofexaminingtheentryofleadaswellascopperandzincintothe
C,SelhorstT,GreinerM,GerofkeA,etal.(2017)
meatofhuntedgamewhenusingeitherleadornon-leadammunition.
Copperandzinccontentinwildgameshotwith
Theoutcomeofthisstudyshowsthattheusageofbothlead-basedammunitionand
leadornon-leadammunition–implicationsfor
consumerhealthprotection.PLoSONE12(9): alternativenon-leadammunitionresultsintheentryofcopperandzincintotheedibleparts
e0184946.https://doi.org/10.1371/journal. ofthegame.Usingnon-leadammunitiondoesnotentaildangerouslyelevatedlevelsofcop-
pone.0184946
perandzinc,soreplacingleadammunitionwithalternativeammunitiondoesnotintroducea
Editor:AntoniMargalida,UniversityofLleida, furtherhealthproblemwithregardtothesemetals.Thelevelsofcopperandzincingame
SPAIN
meatfoundinthisstudyareintherangefoundinpreviousstudiesofgame.Thecontentof
Received:February17,2017 copperandzincingamemeatisalsocomparabletothoseregularlydetectedinmeatandits
Accepted:September2,2017 productsfromlivestock(pig,cattle,sheep)forwhichthemeanhumanconsumptionrateis
muchhigher.Fromtheviewpointofconsumerhealthprotection,theuseofnon-leadammu-
Published:September21,2017
nitiondoesnotposeanadditionalhazardthroughcopperandzinccontamination.Ahealth
Copyright:©2017Schlichtingetal.Thisisanopen
riskduetothepresenceofcopperandzincingamemeatattypicallevelsofconsumerexpo-
accessarticledistributedunderthetermsofthe
CreativeCommonsAttributionLicense,which sureisunlikelyforbothtypesofammunition.
permitsunrestricteduse,distribution,and
reproductioninanymedium,providedtheoriginal
authorandsourcearecredited.
DataAvailabilityStatement:Allrelevantdataare
withinthepaperanditsSupportingInformation
files. Introduction
Funding:Thestudywassupportedmainlybythe Leadornon-lead,thatisthequestion:whetherleadammunitionforhuntingcanorshouldbe
FederalMinistryofFoodandAgriculture(BMEL)in replacedbynon-leadammunition–duetohealthconcernsaboutleadlevelsingamemeat—
Germany(http://www.bmel.de)butalsobythe
hasbeendiscussedintenselyinrecentyears[1,2].Notonlythequestionofapossibleentryof
listedLa¨nder(Mecklenburg-WesternPomerania,
leadintotheediblepartsofgamemeatthroughthedifferentbullettypeshasbeenraised,but
LowerSaxony,Saxony-Anhalt,Bavaria,Hesse,
alsowhethertheothermetalsused(i.e.copperandzinc)enterthemeatinasimilarwayandif
NorthRhine-Westphalia,HamburgandBremen)
andorganisms(EuropeanPoultry,EggandGame so,theirpossiblerelevanceforconsumerhealthprotection[3–5].
PLOSONE|https://doi.org/10.1371/journal.pone.0184946 September21,2017 1/16
CuandZncontentinwildgameshotwithleadornon-leadammunition
Association(EPEGA),DeutscherJagdverbande.V. Non-leadbulletsaresolidbulletsmadeofcopperoralloysofcopperandzinc(tombacor
(DJV,GermanHuntingAssociation),Bayerischer brass),which—dependingontheirconstructionandimpactvelocity–eitherfragmentor
Jagdverbande.V.(BJV,BavarianHunting
expand.Notmuchisknown,however,aboutapossibleincreaseofcopperandzinccontentin
Association),BundesverbandDeutscher
gamemeatthroughtheuseofnon-leadbulletsforhunting[4,6].
Berufsja¨gere.V.(GermanAssociationof
Incontrasttolead,copperandzincareessentialtraceelementsforhumans.Theyare
ProfessionalHunters),VerbandderHerstellervon
Jagd-,SportwaffenundMunitione.V.(JSM, importantpartsofdifferentenzymes,forexample.Nonetheless,aboveacertainconcentration,
AssociationoftheManufacturersofHuntingand copperaswellaszincarealsotoxicaccordingtoParacelsus’observationthat“thedosemakes
SportsWeaponsandAmmunition),Universita¨tfu¨r thepoison”.Copperisstoredintheliverandisexcretedviathebile.Tolerableupperintake
NachhaltigeEntwicklungEberswalde(HNEE,
levelsforcopperare1to5mgperday,andforzinc7to25mgperday,dependingonage[7].
UniversityforSustainableDevelopment)).Alotof
Inordertoobtainaknowledge-basedbackgroundforpoliticaldecisionmaking,the
peoplehelpedobtainingthedata:thehunters,
gametradersandothers,butwerenotemployed researchproject“Safetyofgamemeatobtainedthroughhunting(LEMISI)”wasinitiated[8].
orcontractedtodoso.Thefundershadnoactive TheprojectwasdevelopedinsixregionsinGermanyfrom2011to2014onbehalfoftheFed-
roleinstudydesign,analysis,decisiontopublish eralMinistryofFoodandAgriculture(BMEL).Theeffectsofdifferentbulletmaterials(lead
orpreparationofthemanuscript.
versusnon-lead)onthecontentoflead,copperandzincintheediblepartsofgamemeatwere
Competinginterests:Theauthorshavedeclared examined.
thatnocompetinginterestsexist. WithinthescopeoftheLEMISIproject,theinfluenceofusingalternative(non-lead)
ammunitionontheconcentrationsofcopperandzincingamemeatwasexamined.Thefol-
lowingquestionswereaddressedinthecourseoftheproject:
1. Isthereanydifferenceinthecopperandzinccontentofthegamemeatbetweengame
huntedwithleadammunitioncomparedtonon-leadammunition?
2. Ishighercopperandzinccontentmeasuredintheareaaroundthewoundchannelofani-
malskilledwithnon-leadammunition?
3. Aretheresignificantdifferencesinthecopperandzinccontentinthethreesubsamples
takenfromtheedibletissueofhuntedgame(i.e.,theareaclosetothewoundchannel,the
saddleandthehaunch)?
Previousexperiencesshowthatleadammunitiononaverageresultsinahigherleadcontent
ingamemeatthannon-leadammunition[9,10].Inthefollowing,thedatagainedonthecop-
perandzinccontentinediblemeatarepresentedanddiscussedinordertoavoidreplacing
oneproblemwithanother.
Materialandmethods
Withinthescopeofthestudy,samplesof1254roedeer,854wildboarand90reddeerfrom
differentregionswithinGermanywereexamined[8].
Ethicsstatement
Licensedhunterskilledthegameanalysedinthisstudyduringtheestablishedhuntingseason
andinaccordancewithGermanregulations(GermanHuntingAct;Bundesjagdgesetz)and
bestpractices.Itdidnotinvolveanyadditionalkillingotherthanwhatiscarriedoutinthe
Germanforestsonaregularandmanagerialbasis(populationcontrol).Permissionwas
grantedfromtheGermanFederalStates(La¨nder)andtheirrespectivehuntingauthorities.
Choiceofregions
WithinGermanysixregionswerechosenaccordingtotheleadcontentofthetopsoilinorder
tocontrolleadconcentrationsattributabletosoilleadcontaminationinthe(statistical)analy-
sis.Tworegionswereselectedforeachofthethreeleadlevelsintopsoil(i.e.lowleadcontent:
<30mglead/kgsoil,mediumleadcontent:30to75mglead/kgsoilandhighleadcontent:>
PLOSONE|https://doi.org/10.1371/journal.pone.0184946 September21,2017 2/16
CuandZncontentinwildgameshotwithleadornon-leadammunition
75mglead/kgsoil)chosenusingageographicalmapindicatingleadcontentintopsoil,thus
resultinginatotalofsixregions[11].Thecontentofcopperandzincinsoilwerenottaken
intoaccountduetotheheterogeneityofsoilconditionsandthemovementofanimals.
Experimentaldesignandimplementation
Qualityassurancemeasureswereintegratedinallphasesoftheproject.Hunterswereinstructed
astotheaimsoftheresearchproject.Theanimalswereeithershotwithspecificleadammuni-
tionorwithspecificnon-leadammunition.Foreachanimalkilled,thehuntershadtofillina
sampledatasheetinwhichdetailedinformationontheanimals(species,ageandgender)and
howtheyhadbeenshot(includingbulletmaterial,i.e.leadvsnon-lead),bullettypeused,infor-
mationontheentryandexitofthebullet,shootingdistance,bonehit(i.e.iftheanimalwas
killedbyashotthatthehunterreportedtohavestrucknotonlytissueandorgansbutalsoskele-
talstructuressuchastheribs,scapula)wererecorded.Parametersincludedinthestatistical
analysisweretheanimalspeciesandbulletmaterial—leadammunitionversusnon-leadammu-
nition.Theentryandexitofthebulletwereconsideredinordertodiscussthedistributionof
themetalsinthemeatdependingontheplaceofentry.Inaddition,socalledbone-hits(see
above)werealsoexamined.Here,theunderlyinghypothesisisthattheresistanceofthebone
couldleadtoafurtherdistributionofthemetalsinthemusclecomparedtobullethitsof“softer
tissues”.Thesampledatasheetwasalsoavitalpartoftheoverallqualityandassurancecontrol
(seebelow).
Thehuntedgamewasbroughttogametraderswhohadalsobeenspecificallytrainedfor
thisprojectandwhocollectedthesamplesaccordingtouniformstandards.Threesamples
weretakenfromeachanimalaftercompletionoftheregularprocessofskinningandcleaning
thecarcassaccordingtohygienestandardsforgamemeat[12].Thesamplesweretakenfrom
marketablemeatofthesaddle,haunchandtheareaclosetothewoundchannel,whichhad
beenwidelycutout.Thesampleamountwas100gforeachofthethreesubsamples.Subsam-
pleswerestoredincolouredvials(i.e.onecolourforeachtypeofsubsample).Sampleswere
numberedandcoded.Allthreesubsamplesperanimalwerestoredinvialsinpolythenebags.
Thecorrespondingsampledatasheet(withtheidenticalcoding)wasstoredinaseparatepoly-
thenebag.Thesetwobagswerestoredtogetherinathirdpolythenebagsothatitwaspossible
totracebackeachsubsampletothelocationwheretheanimalwasshot,thelaboratorywhere
analyseswereconductedandalltheotherrelevantparametersgiveninthesampledatasheet.
Inthisway,thissystemservedasqualityassuranceandcontrol(i.e.plausibilitycheck).Until
thetimeofchemicalanalysis,sampleswerefrozenandstoredinpolythenebagsat−18C.
Analytics
Thesamplesweretransportedto12accreditedlaboratoriesforchemicalanalysis:11of
themfromgovernmentalagenciesandonebelongingtoaleadinginternationalgroupof
laboratories.
Beforethebeginningofchemicalanalysis,thesampleswerehomogenizedand0.5to1gof
eachsamplewasputinahigh-pressureTefloncontainerformicrowavepressuredigestionin
linewithEN13805:2014[13].Thecontentofcopperinmusclesampleswasdeterminedeither
byusingtheinductivelycoupledplasma–massspectrometricmethod(ICP-MS),byapplying
inductivelycoupledplasmaopticalemissionspectrometry(ICP-OES)oralternatively,by
applyinggraphitefurnaceatomicabsorptionspectrometry(GFAAS)[14–16].Thezinccontent
inmusclesampleswasdeterminedeitherwithICP-MS/ICP-OESoralternatively,byapplying
flameatomicabsorptionspectrometry(FAAS)[17].
PLOSONE|https://doi.org/10.1371/journal.pone.0184946 September21,2017 3/16
CuandZncontentinwildgameshotwithleadornon-leadammunition
Determinationofplausibility
TheanalyticalresultsweresenttotheEberswaldeUniversityforSustainableDevelopment
(Hochschulefu¨rnachhaltigeEntwicklungEberswalde,HNEE)foraplausibilitycheckofthe
huntingandbulletdatausingthenumericcodingofsamplesfromthelaboratoriesandthe
completeinformationfromthedatasheets.Themostimportantitemwasthecorrectidentifi-
cationofthebulletsusedasreportedbythehuntersinthesampledatasheetsas“lead”or
“non-lead”.TheapproveddataweresubsequentlysenttotheGermanFederalInstitutefor
RiskAssessment(Bundesinstitutfu¨rRisikobewertung,BfR)wherethestatisticaldataanalyses
aswellasthetoxicologicalriskassessmentwereperformed.
Statisticalevaluation
Thecopperandzinccontentwerequantifiableinallexaminedsubsamples.Sincethedata
werenotdistributednormallyandthedistributionswerehighlyheterogeneous,groupcom-
parisonsweredoneusingnon-parametricmethods[18].TheMann-WhitneyUtestwas
appliedwhencomparingleadshotsampleswithnon-leadshotsamples.Thecomparisonof
thesubsampleswasmadebyapplyingeithertheFriedmantestortheWilcoxonsigned-rank
test.Thesignificancelevelwasdeterminedasp<0.05.Whencomparingthesubsamples,mul-
tipletestingwastakenintoaccountusingacorrespondingBonferroni-adjustedsignificance
level(p<0.017)[19].
Thedistributionoftheanalyticalresultsisdisplayedgraphicallyusingbeanplots(R-package
“beanplot”[20]).Beanplotsconstituteanalternativetoboxplots.Theycombineadensity
shapewithaone-dimensionalscatterplot–showingallanalyticaldataassmalllines–thereby
allowingavisualcomparisonofthedistribution[21].
StatisticalanalysiswererealizedusingSPSS(IBMSPSSStatisticsforWindows,Version
21.0).CorrespondinggraphswerecreatedusingR[22].
Results
Copper
Themajorpartoftheobservedcoppercontentinroedeer,wildboarandreddeerwhichhad
beenhuntedusingnon-leadammunitionwasinalowrange.Thisfactisunderlinedbythe
heightofthe95thpercentiles(Table1),aswellasbythedistributionofcoppercontentinthe
beanplots(Fig1andS1Fig).
Theaveragecoppercontentofthesamplesofnon-leadshotroedeerwashigherthanthatof
leadshotroedeer.Thusthecoppercontentclosetothewoundchannelwassignificantlydiffer-
entdependingonthetypeofammunition(Mann-WhitneyUtest:p<0.0001;Table1).Butthe
samplesfromtheareaclosetothewoundchannelofnon-leadshotroedeershowedsignifi-
cantlylowercoppercontentthansamplesfromthehaunchorsaddle(Wilcoxonsigned-rank
testeach:p<0.0001;Table1).Fortheroedeersamples,thehighestcoppercontentwasdetected
inasampleofthesaddle(Table1).
Overall,samplesof14roedeerhadcoppercontentabove5mg/kg.Thereof,13roedeer
wereshotwithnon-leadammunition.Twelveoftheseanimalswerekilledwitha“bonehit”
(fordefinitionseematerialandmethodssection).Oneanimalshotwithnon-leadammunition
andkilledwithbonehithadincreasedcoppercontentinsamplesbothfromtheareaaround
thewoundchannel(9.70mg/kg)andfromthehaunch(9.05mg/kg).
Forwildboar,thesamplesfromtheareaclosetothewoundchannelandthesaddleshowed
significantlyhighercoppercontentwhennon-leadammunitionhadbeenused(Mann-Whit-
neyUtesteach:p=0.005).Nevertheless,thehighestcoppercontentinwildboarsampleswas
PLOSONE|https://doi.org/10.1371/journal.pone.0184946 September21,2017 4/16
CuandZncontentinwildgameshotwithleadornon-leadammunition
Table1. Coppercontentinhuntedroedeer,wildboarandreddeer(mg/kg).
Sample Bullet N Meana Median 95thb Maximum P
Roedeer,haunch Lead 745 1.614 1.564 2.196 6.451 0.359
Non-lead 509 1.695 1.577 2.702 9.048
Roedeer,saddle Lead 745 1.810 1.759 2.769 4.034 0.576
Non-lead 509 2.017 1.730 3.672 37.537
Roedeer,aroundwoundchannel Lead 745 1.464 1.400 2.063 3.946 <0.0001
Non-lead 509 1.635 1.500 2.444 9.701
Wildboar,haunch Lead 514 1.437 1.375 2.136 4.300 0.432
Non-lead 340 1.456 1.368 2.363 8.050
Wildboar,saddle Lead 514 1.506 1.200 1.986 110.000 0.005
Non-lead 340 1.404 1.270 2.420 5.238
Wildboar,aroundwoundchannel Lead 514 1.426 1.322 2.286 9.616 0.005
Non-lead 340 1.627 1.419 2.728 18.886
Reddeer,haunch Lead 64 1.891 1.857 2.648 2.969 0.954
Non-lead 26 1.896 1.874 2.478 2.902
Reddeer,saddle Lead 64 1.794 1.746 2.462 4.787 0.789
Non-lead 26 1.759 1.760 2.280 2.390
Reddeer,aroundwoundchannel Lead 64 1.701 1.743 2.165 2.553 0.712
Non-lead 26 1.755 1.650 2.363 2.721
aArithmeticalmean.
b95thpercentile.
https://doi.org/10.1371/journal.pone.0184946.t001
foundinasamplefromthesaddleofananimalwhichhadbeenshotwithleadammunition
(Table1andFig1).
Whenusingnon-leadammunition,thecoppercontentintheareaclosetothewound
channelinwildboarsampleswashigherthanthatofthehaunch(Wilcoxonsigned-ranktest:
p=0.002)orsaddle(Wilcoxonsigned-ranktest:p<0.0001).Forleadshotanimals,thesamples
fromtheareaclosetothewoundchannelshowedsignificantlyhighercoppercontentthan
samplesfromthesaddle(Wilcoxonsigned-ranktest:p<0.0001),buttheywerestillbelowthe
coppercontentofthesamplesfromthehaunch(Wilcoxonsigned-ranktest:p=0.008).
Thecoppercontentofatotalof12wildboarsampleswasaboveavalueof5mg/kg.Of
these,fouranimalswereshotusingnon-leadammunitionandsevenanimalsusinglead
ammunition.Fromtheseanimals,eight(ninesamples)hadbeenkilledbyabonehit(non-
lead:fivesamples;lead:foursamples).Inoneanimalwhichbeenshotusingnon-leadammuni-
tion,thesamplefromthehaunchaswellasthesamplefromtheareaclosetothewoundchan-
nelhadincreasedcoppervalues(haunch8.05mg/kgandareaclosetothewoundchannel7.55
mg/kg,bonehit).
Thecomparisonofthecoppercontentforreddeershowednosignificantdifferences
betweentheuseofnon-leadorleadammunition(Table1).
Acomparisonbetweenroedeerandwildboarshowedthatthecoppercontentofroedeer
washigherthanthatofwildboar(Mann-WhitneyUtest;Table2)irrespectiveofthesubsam-
pleandtypeofammunitionused.
Zinc
Thezinccontentinthesamplesofroedeeraswellasinthoseofwildboarvariedconsiderably,
butextremevalueswereonlysporadicallyfound(Fig2andS2Fig).
PLOSONE|https://doi.org/10.1371/journal.pone.0184946 September21,2017 5/16
CuandZncontentinwildgameshotwithleadornon-leadammunition
Fig1.Coppercontentindifferentediblepartsofroedeerandwildboarbybulletmaterial(lead,non-lead).
https://doi.org/10.1371/journal.pone.0184946.g001
Thezinccontentinroedeersamplesfromtheareaclosetothewoundchannelwassignifi-
cantlyhigherwhenusingnon-leadammunitioncomparedtoleadammunition(Mann-Whitney
Utest,p<0.0001).Inaddition,thezinccontentinsamplesfromthesaddlewassignificantly
higherwhenusingnon-leadammunition(Mann-WhitneyUtest,p=0.006),butthemedianval-
ueswereonlyslightlydifferent.Thisdifferencecanbeseenbylookingatthe95thpercentile
(Table3),aswellasoveralldistribution(Fig2).Regardlessofthetypeofammunition,theroedeer
samplesfromtheareaclosetothewoundchannelwerenotsignificantlydifferentfromthose
fromthehaunchorsaddle(Friedmantest;non-lead:p=0.281,lead:p=0.149,respectively).
PLOSONE|https://doi.org/10.1371/journal.pone.0184946 September21,2017 6/16
CuandZncontentinwildgameshotwithleadornon-leadammunition
Table2. Differencesincoppercontentofdifferenttissuesfromroedeerandwildboarbybulletmaterial(Mann-WhitneyUtest).
Sample Bullet Species N Meana Median P
Haunch Lead Roedeer 745 1.614 1.564 <0.0001
Wildboar 514 1.437 1.375
Non-lead Roedeer 509 1.695 1.577 <0.0001
Wildboar 340 1.456 1.368
Saddle Lead Roedeer 745 1.810 1.759 <0.0001
Wildboar 514 1.506 1.200
Non-lead Roedeer 509 2.017 1.730 <0.0001
Wildboar 340 1.404 1.270
Aroundwoundchannel Lead Roedeer 745 1.464 1.400 <0.0001
Wildboar 514 1.426 1.322
Non-lead Roedeer 509 1.635 1.500 0.0010
Wildboar 340 1.635 1.419
aArithmeticalmean.
https://doi.org/10.1371/journal.pone.0184946.t002
In171roedeersamples,thezinccontentwasabove50mg/kg(101ofthesesampleswere
shotusingnon-leadammunition).Ofthese171roedeersamples,129sampleswerebonehits
(non-lead:79samples,lead:50samples).
Samplesofwildboaralsohadsignificantlyhigherzinccontentintheareaclosetothe
woundchannelwhenusingnon-leadammunition(Mann-WhitneyUtest:p=0.027).
Thezinccontentofsamplesfromthesaddleofwildboarwassignificantlyhigherwhenusing
leadammunitionascomparedtonon-leadammunition(Mann-WhitneyUtest,p=0.049).
Whencomparingthesubsamplesfromwildboarshotwithnon-leadammunition,thezinccon-
tentofsamplesfromtheareaaroundthewoundchannelweresignificantlyhigherthanthoseof
thesamplesfromthesaddle(Wilcoxonsigned-ranktest:p<0.0001).Thezinccontentofsamples
fromtheareaclosetothewoundchannelwerealsohigherthanthoseofthehaunch,butthey
didnotdiffersignificantly(Wilcoxonsigned-ranktest:p=0.591).Whenleadammunitionwas
used,thezinccontentinsamplesfromtheareaclosetothewoundchannelwaslowerthanthe
zinccontentofsamplesfromthehaunch(Wilcoxonsigned-ranktest:p<0.0001).Thezinccon-
tentofsamplesfromtheareaclosetothewoundchannelandfromthesaddlewerenotsignifi-
cantlydifferent(Wilcoxonsigned-ranktest:p=0.048).
Thezinccontentof111wildboarsampleswasabove50mg/kg(ofthese63camefrom
non-leadshotanimals).Furthermore,78ofthesesampleswerefromanimalskilledbybone
hits(non-lead:48,lead:30).
Justasforthecoppercontent,thezinccontentofreddeersamplesshowednosignificant
differencesbetweennon-leadandleadammunition.
Whencomparingsamplesofroedeerandwildboar,asignificantdifferenceintheirzinc
contentcanonlybeseenforsamplesofthesaddlewhenusingnon-leadammunition.Thezinc
contentofsamplesfromthesaddleofroedeerissignificantlyhigherthanthoseofwildboar
(Mann-Whitney-U-test,Table4).
Discussion
OneoftheaimsoftheLEMISIprojectwastodeterminepossibledifferencesinthecopper
andzinccontentingamemeatoftheexaminedspecieswhenusingleadornon-leadammuni-
tionforhunting.Bothtypesofammunitioncontaincopperandzinc.Whereasnon-leadbul-
letsaremainlycopper-zincalloyswithpartlydifferingcoppercontent,manylead-based
PLOSONE|https://doi.org/10.1371/journal.pone.0184946 September21,2017 7/16
CuandZncontentinwildgameshotwithleadornon-leadammunition
Fig2.Contentindifferentediblepartsofroedeerandwildboarbybulletmaterial(lead,non-lead).
https://doi.org/10.1371/journal.pone.0184946.g002
bulletsusedforhuntingaresurroundedbyatombacjacket,whichhasahighcopper(>80%)
andzinccontent.Forbothmetals,variationsinamountcouldbeobservedforleadandnon-
leadammunition.
Themaximumresiduelevel(MRL)forcopperpermittedinfoodofanimaloriginfrom
pigs,cattle,sheep,goats,horses,poultryandotherfarmanimalsis5mg/kg(freshweight)
accordingtoregulation(EC)No149/2008andtheamendingregulation(EC)No396/2005.
Thisregulationappliestoallresiduesofpesticides,veterinarydrugs,orbiocidesinoronfood
andfeedofplantandanimalorigin.Forwildgamemeat(i.e.themeatafterremovalof
PLOSONE|https://doi.org/10.1371/journal.pone.0184946 September21,2017 8/16
CuandZncontentinwildgameshotwithleadornon-leadammunition
Table3. Zinccontentinhuntedroedeer,wildboarandreddeer(mg/kg).
Sample Bullet N Meana Median 95thb Maximum P
Roedeer,haunch Lead 745 30.574 31.660 44.640 65.000 0.089
Non-lead 509 31.946 32.000 48.000 64.000
Roedeer,saddle Lead 745 28.842 31.324 50.000 63.000 0.006
Non-lead 509 31.348 31.770 55.800 131.584
Roedeer,aroundwoundchannel Lead 745 30.532 29.719 48.000 72.296 <0.0001
Non-lead 509 33.649 32.870 53.624 138.000
Wildboar,haunch Lead 514 31.700 32.029 45.700 56.000 0.397
Non-lead 340 31.358 31.000 49.407 70.073
Wildboar,saddle Lead 514 28.266 29.000 45.000 98.521 0.049
Non-lead 340 27.646 25.975 52.168 95.202
Wildboar,aroundwoundchannel Lead 514 30.406 28.410 52.000 88.232 0.027
Non-lead 340 32.360 30.919 55.955 78.036
Reddeer,haunch Lead 64 33.965 35.216 43.225 52.642 0.302
Non-lead 26 35.850 36.373 52.410 57.510
Reddeer,saddle Lead 64 35.371 37.486 53.010 58.990 0.689
Non-lead 26 35.134 31.569 63.580 74.640
Reddeer,aroundwoundchannel Lead 64 32.992 31.450 48.030 70.457 0.715
Non-lead 26 34.110 32.575 48.417 67.933
aArithmeticalmean.
b95thpercentile.
https://doi.org/10.1371/journal.pone.0184946.t003
trimmablefat)thepermittedresiduelevelsofarhasbeen0.01mg/kg,whichcorrespondswith
thelowerlevelofdetection.Thisisbecausesincespring2013“gamemeat”hasbeenlisted
under“otherterrestrialanimalproducts”inAnnexItoregulation(EC)No212/2013andthe
amendingregulation(EC)No396/2005andnoresiduevaluehasbeenderivedbasedonnatu-
ralcontentuptonow.
Inordertoaccountforthenaturalbackgroundlevelsofcopperingamemeat(asaresultof
environmentaluptakemainlythroughfeeding),Germany–initsroleas“evaluatingmember
Table4. Differencesinzinccontentofdifferenttissuesfromroedeerandwildboarbybulletmaterial(Mann-WhitneyUtest).
Sample Bullet Species N Meana Median P
Haunch Lead Roedeer 745 30.574 31.660 0.1330
Wildboar 514 31.700 32.029
Non-lead Roedeer 509 31.946 32.000 0.3360
Wildboar 340 31.358 31.000
Saddle Lead Roedeer 745 28.842 31.324 0.3040
Wildboar 514 28.266 29.000
Non-lead Roedeer 509 31.348 31.770 <0.0001
Wildboar 340 27.646 25.975
Aroundwoundchannel Lead Roedeer 745 30.532 29.719 0.3330
Wildboar 514 30.406 28.410
Non-lead Roedeer 509 33.649 32.870 0.0970
Wildboar 340 32.360 30.919
aArithmeticalmean.
https://doi.org/10.1371/journal.pone.0184946.t004
PLOSONE|https://doi.org/10.1371/journal.pone.0184946 September21,2017 9/16
CuandZncontentinwildgameshotwithleadornon-leadammunition
Fig3.95thpercentilecoppercontentoffarmanimals(Germanfoodmonitoringprogram)andgame
meat(LEMISI)aswellastheacceptablemaximumresiduelevelofcopperinfarmanimals.Redbroken
line:5mgcopper/kgmeat.
https://doi.org/10.1371/journal.pone.0184946.g003
state”—proposedaresiduelevelforcopperingamemeatof4mg/kg[23].Theproposedvalue
isderivedfromGermanfoodmonitoringdata[24],andincorporatesthe95thpercentileofthe
determinedcoppercontent.EFSAfoundthatthecontributionoftheproposedMRLtototal
consumerexposuretocopperwasnegligible.Itamountsupto0.7%oftheAcceptableDaily
Intake(ADI)ofanadult[23].ThisfactrecommendsthesettingoftheMRLat4mg/kgfor
coppercompoundsinwildgameinordertocoverthenaturalbackgroundlevelofcopper
observedinthesurveyconductedinGermanyin2012.Itshouldbenotedthatthegamemeat
examinedforthemonitoringhadbeenshotusingleadammunition.
Themaximumresiduelevelsmentionedabovecanbeusedasgeneralguidancesincethe
resultsobtainedwithinthescopeoftheLEMISI-projectshowthatcopperisnotevenlydistrib-
utedinthegamemeat.Thedataindicatethatanexceedanceofthemaximumresiduelevels
forcopperingamemeatcannotbeexcludedandthatthevarianceofthecoppercontent
detectedisratherlarge.AsshowninTable1,themaximumresiduelevelwasexceeded,in
somecasesmultipletimes,inallexaminedsubsamples(i.e.haunch,saddle,meatclosetothe
woundchannel)whenusingeitherleadornon-leadammunitionforhunting.Onesampleof
roedeer(saddle,non-leadammunition)hadacoppercontentof37.5mg/kg,andonesample
ofwildboar(saddle,leadammunition)hadacoppercontentof110.0mg/kg.
Theresultsofthecoppercontentindifferentmeatsamplesdonotpresentaconsistentpic-
ture.Regardingthe95thpercentile,itcanbeseenthatthecoppercontentisslightlyhigherinthe
areaclosetothewoundchannelthaninthesaddleorhaunchwhenusingnon-leadammunition
forhuntingwildboar.Ontheotherhand,thecoppercontentmeasuredinroedeersamplesof
theareaclosetothewoundchannelislowerthaninsamplesofhaunchandsaddlewhenusing
eithernon-leadorlead-basedammunition.However,someofthesecontradictoryfindingsfor
copperandzinccouldalsobearesultofthesamplesize,whichmaynothavebeensufficientfor
someofthesubgroupsanalysed,eventhoughoverallitwasquiteaconsiderablesamplesize.
Themedianvaluesofthecoppercontentofleadornon-leadshotgamemeatwererelatively
closetogethertoalargeextent.Leadaswellasnon-leadbulletsresultinacomparableentryof
copperintotheediblepartsofthegamewithonlyminordifferences.Comparingthe95thper-
centilesofcoppercontentinediblemeatofpork,vealandbeefwiththe95thpercentileofthe
coppercontentofsamplesofroedeer,wildboarandreddeer,itbecomesapparentthatpork
hasthelowestcoppercontent,whereasthepercentilevaluesforbeefandaboveallvealareina
rangecomparabletogamemeat(Fig3).
Thelevelsofzincattributabletotheuseofleadammunitionareslightlyhigherinsomesub-
samplesascomparedtothelevelsofzincwhenshootingwithnon-leadammunition.This
PLOSONE|https://doi.org/10.1371/journal.pone.0184946 September21,2017 10/16