Turk.J.Vet.Anim.Sci. 2007;31(1):39-45 ©TÜB‹TAK ResearchArticle AcuteToxicityofCarbaryl,Methiocarb,andCarbosulfan totheRainbowTrout(Oncorhynchusmykiss)andGuppy (Poeciliareticulata) MuhammetBORAN*,‹lhanALTINOK,ErolÇAPKIN,HikmetKARAÇAM,VeyselB‹ÇER FacultyofMarineSciences,KaradenizTechnicalUniversity,61530Sürmene,Trabzon-TURKEY Received:31.01.2006 Abstract: Theacutetoxicityofcarbaryl,methiocarbandcarbosulfantojuvenilerainbowtrout( Oncorhynchusmykiss;1.92±0.5 g)andguppies(Poeciliareticulata;0.45±0.04g)wasevaluatedin96-htoxicitytestsunderstaticconditions.Concentrationsinthe toxicitytestrangedfrom0.85to8.5mg/lforcarbaryl,from0.7to28.0mg/lformethiocarb,andfrom0.031to12.5mg/lfor carbosulfan.Carbarylandmethiocarbweremoretoxictorainbowtroutthantoguppies.However,comparedwithrainbowtrout, guppiesweremoresensitivetocarbosulfan.Theincidenceofmortalityinfishincreasedsignificantly(P<0.05)withincreasi ng pesticideconcentrations.Thetimerequiredtokill50%(LT 50)oftherainbowtroutatthelowestconcentrationsofcarbaryl, methiocarb,andcarbosulfanwas51h12min,74h35min,and107h57min,respectively.Asinsecticideconcentrationswere increased,LT50 valuesdecreased(P<0.05). KeyWords: Acutetoxicitytest,carbamate,pesticides,rainbowtrout,guppies Karbaril,MetiyokarbveKarbosülfan’›nGökkufla¤›Alabal›¤›( Oncorhynchusmykiss)veLepistes (Poeciliareticulata)Bal›klar›ÜzerineAkutToksikEtkileri Özet: Karbaril,metiyokarbvekarbosülfanaktifmaddeleriniiçereninsektisitleringökkufla¤›alabal›klar›(Oncorhynchusmykiss;1,92 ±0,5g)velepistesbal›klar›( Poeciliareticulata ;0,45±0,04g)üzerine,akuttoksiketkileri,statiktestyöntemi(96saatlik) kullan›larakbelirlenmifltir.Testçözeltilerindekinominalinsektisitmiktarlar›karbariliçin0,85-8,5mg/l,metiyokarbiçin0 ,7-28,0 mg/lvekarbosülfaniçin0,031-12,5mg/lolacakflekildeayarlanm›flt›r.Karbarilvemetiyokarb›ngökkufla¤›alabal›klar›üzerinetoksik etkilerinin,lepistesbal›klar›nagöredahafazlaoldu¤utespitedilmifltir.Ancaklepistesbal›klar›n›nkarbosülfanaktifmadde sinekarfl›, alabal›klaragöredahahassasolduklar›belirlenmifltir.Testedilenbal›klardakiölümoranlar›n›n,akvaryumlardakiçözeltileri çersinde bulunanaktifmaddelerinkonsantrasyonunaba¤l›olarakönemliderecede(P<0,05)artt›¤›saptanm›flt›r.Endüflükkarbaril, metiyokarbvekarbosülfankonsantrasyonlar›namaruzb›rak›langökkufla¤›alabal›klar›n›n%50’sininölmesiiçingerekenzaman s›ras›yla51saat12dakika,74saat35dakikave107saat57dakikaolaraktespitedilmifltir.‹nsektisitkonsantrasyonuartt›kçaLT50 de¤erinindüfltü¤ügörülmüfltür(P<0,05). AnahtarSözcükler: Akuttoksisitetesti,karbamat,pestisit,gökkufla¤›alabal›¤›,lepistes Introduction environment,andtheytendnottobioaccumulate. Carbamatepesticidesareusedwidelyforagricultural However,carbamatepesticidesaretoxictonon-targeted andresidentialapplicationsasinsecticidesandfungicides. wildlifeandfishandbirdsappeartobethemoresensitive Theirworldwideannualuseisestimatedtorangefrom thanmammalstothesepesticides(1). 20,000to35,000t.Thisfamilyofchemicalsreplacedthe Thecarbamateesterderivativesusedasinsecticides organochlorinepesticides,whichhavebeenbanned aregenerallystableandhavealowvaporpressureand throughouttheworld.Unlikeorganochlorinepesticides, lowwatersolubility.Althoughcarbamatesarenotvery carbamateinsecticidesdonotpersistlonginthe stableunderaquaticconditionsandwillnotpersistlong *E-mail:[email protected] 39 AcuteToxicityofCarbaryl,Methiocarb,andCarbosulfantotheRainbowTrout( Oncorhynchusmykiss)andGuppy(Poeciliareticulata) intheenvironment,bioaccumulationtosomeextent MaterialsandMethods occursinfishduemainlytotheirslowmetabolism(2). ExperimentalAnimals Pesticidesthataresolubleinwaterareeasilytransported intosurfacewaters.Pesticidesadsorbedintosoilparticles Rainbowtrout(Oncorhynchusmykiss;1.92±0.5g) canbecarriedfromapplicationsitesonsediment.The wereobtainedfromKTU,FacultyofMarineSciences, runoffwatercandissolvetheactiveingredients,which ResearchFacilityinTrabzon,Turkey.Thefishwere aresubsequentlytransportedtounintendedsites.Either acclimatedtolaboratoryconditionsinflowthrough dissolvedinwaterorcarriedbysediment,pesticidesthat fiberglasstank(100l)underanaturalphotoperiodfor2 arecarriedoff-sitecancontaminatesurfacewaters(3,4). weeks.Duringtheacclimationperiod,thefishwerefed5 timesadaywithcommercialfishfeedat5%body Otherthantargetedpests,insecticidescanaffectnon- weight.Guppies(Poeciliareticulata;0.45±0.04g)were targetedspeciessuchasaquaticanimals.Ingeneral, obtainedfromalocalfishstoreandacclimatedto methiocarbismoderatelytoxictoaquaticorganismsand laboratoryconditionsinglassaquaria(50l)filledwith thetoxicdosedependsonspecies(5).Thetoxicityof agedtapwaterunderanaturalphotoperiodfor2weeks. carbamatesubstancesresultsprimarilyfrominhibitionof Thefishwerefedtwiceadaywithcommercialflakesand acetylcholinesterase(AChE),akeyenzymeofthenervous onceaweekwithDaphnia sp. system.Theinhibitioncausesanaccumulationof ExperimentalDesign acetylcholineinsynapseswithdisruptionofthenerve Afteracclimation,therainbowtroutwererandomly function,whichcanresultindeath(6). transferredto20-laquariaandtheguppiesto5-laquaria. Therearelimitednumbersofstudiescomparingthe Theexperimentswereperformedinstaticwater(10fish acutetoxicityofcarbaryl,carbosulfanandmethiocarbin peraquarium).Testsolutionswerepreparedfrom variousfishes,whileinformationonthemechanismsof commercialformulations(BayerCropScienceAG, toxicitytodifferentfishspecieshasbeenreportedoften Frankfurt,Germany)containing85%(activeingredient) (2,6).Carbarylcancauseadecreaseintheliverglutamine carbaryl,50%methiocarb,and25%carbosulfan.The level(7),andadecreaseinthenumberofleukocytesin concentrationstestedforcarbarylwere0(control),0.85, Gorragothylagothyla (8).Incarbaryltoxicity, 1.53,2.72,4.76,and8.5mg/l.Themethiocarb characteristicbehavioralchangesassociatedwithchanges concentrationswere0(control),0.7,1.4,2.8,5.0,9.0, inphysiologicalparametersinducedbycarbarylexposure 16.0,and28.0mg/l.Therainbowtroutwerenotexposed canbeseen.Histologically,methiocarb-exposedfishhad to0.7or1.4mg/lmethiocarb.Theconcentrationstested forcarbosulfanwere0(control),0.125,0.625,1.25, lamellaredema,separationofepidermisfromlamellae, 6.25,12.5mg/lforrainbowtrout,withadditional0.031 lamellarfusion,telangiectasis,swellingoftheepithelial and0.063mg/ltreatmentsforguppies.Triplicateaquaria cellsandincreasedcytoplasmicgranularity.Gillsalsohad weredesignatedforeachconcentration.Carbaryland scatteredareasoffocallamellarhyperplasia.Methiocarb- methiocarbweredissolvedin10mlofethanol,andthen exposedfishhadnecrosisbetweenthemolecularand addedtotheaquaria.Thecontroltanksreceived10mlof granularlayerofcerebellumwherePurkinjecellsare distilledwater.Sincecarbosulfanwasanemulsifiable present(9). concentrate,itwasdirectlyaddedtotheaquaria.This Thepresentstudysoughttodeterminetheacute studywasconductedunderOECDGuidelineNo.203 toxicityofcommonlyusedpesticidescarbaryl, understatictestconditions(10).Duringthe96-hacute methiocarb,andcarbosulfantorainbowtrout toxicityexperiment,thewaterineachaquariumwas (Oncorhynchusmykiss )andguppies( Poeciliareticulata ) aeratedandhadthefollowingcharacteristicsforrainbow usingastatictestsystem.Asstudiesonthelethaltimes trout:dissolvedoxygen9.40±0.3mg/l,temperature 15.3±0.9°C,pH7.40±0.21,totalhardness99±5 ofcarbaryl,methiocarb,andcarbosulfanforrainbow mg/lasCaCO ,andalkalinity87±3mg/lasCaCO .The troutandguppyintheliteraturearelimited,theother 3 3 watercharacteristicsfortheguppiesweredissolved objectiveofthisstudywastodeterminetheacutetoxicity oxygen8.58±0.2mg/l,temperature21±0.5°C,pH ofcarbaryl,methiocarb,andcarbosulfanconcentrationto 7.35±0.11,totalhardness95±1mg/lasCaCO ,and rainbowtroutandguppiesover96h. 3 40 M.BORAN,‹.ALTINOK,E.ÇAPKIN,H.KARAÇAM,V.B‹ÇER alkalinity75±2mg/lasCaCO 3.Fishwereconsidered exposure.Thetoxicityofcarbaryl,methiocarb,and deadwhengilloperculaandbodymovementceased,and carbosulfantorainbowtroutandguppiesincreasedwith thedeadfishwereremovedimmediately.Theincidenceof increasingpesticideconcentrationanddurationof fishmortalitywasrecorded0,2,3,4,5,6,7,8,9,10, exposure.Carbarylandmethiocarbwerefoundtobe 11,12,24,36,48,60,72,84,and96hafterexposure moretoxictorainbowtroutthantoguppies,while toinsecticides. guppiesweremoresusceptibletocarbosulfanthan WaterQuality rainbowtrout(Tables1-4).Themortalityrateinfish increasedsignificantlywithincreasingconcentrationsof Duringexposuretocarbamateinsecticideswater pesticides.Thecarbarylconcentrationslethalto50%of qualitycharacteristics(temperature,dissolvedoxygen, therainbowtroutwere1.28mg/l(24h),1.14mg/l(48 pH,totalhardness,andalkalinity)ineachtreatmentwere h),1.02mg/l(72h),and0.85mg/l(96h)(Table1). measureddaily.Totalhardnessandtotalalkalinitywere measuredbytitration(11).Temperatureanddissolved Afterexposingrainbowtrouttodifferent oxygenconcentrationsweredeterminedwithanYSIB51 concentrationsofinsecticidesfor24h,theconcentrations polarographicoxygenmeter,andthermistor(Yellow lethalto10%(LC10)and90%(LC90)differedbyafactor SpringInstrument,YellowSpring,Ohio,USA).WaterpH of2.06,2.06,and9.83forcarbaryl,methiocarband wasdeterminedwithaglasselectrode(ThermoOrion, carbosulfan,respectively(Table1).LC 10 valuesof Beverly,Massachusetts,USA). carbarylwere5.88times,ofmethiocarbwere4.95 times,andofcarbosulfanwere3.32timeshigherthan StatisticalAnalyses theLC 90 valuesforguppies(Table3).Therewere Statisticaltestanalysiswasdescribedpreviouslyby significantdifferencesinnumbersofdeadfishbetween Altinok(12).Briefly,theconcentrationofinsecticidesand thedurationof1and96hineachconcentration(P< lethaltimevaluesestimatedtokill10%,50%,and90% 0.05).Thehighestconcentrationofinsecticidescaused ofrainbowtroutandguppieswithin6(6-hLC50),12,24, thehighestfishmortality.Significantdifferenceswere