Assessment of genotoxic activity of petroleum hydr

EcotoxicologyandEnvironmentalSafety62(2005)415–420Assessmentofgenotoxicactivityofpetroleumhydrocarbon-bioremediatedsoilGraz’ynaPazaa,
,GrzegorzNa˛ecz-Jaweckib,KrzysztofUlfiga,RobinL.BrigmoncaInstituteforEcologyofIndustrialAreas,40-844Katowice,Kossutha6Street,PolandbDepartmentofEnvironmentalHealthSciences,MedicalUniversity,Banacha1Street,02-097Warszawa,PolandcSavannahRiverNationalLaboratory,Aiken,SC29808,USAReceived10February2004;receivedinrevisedform29September2004;accepted27October2004Availableonline8January2005AbstractTherelationshipbetweentoxicityandsoilcontaminationmustbeunderstoodtodevelopreliableindicatorsofenvironmentalrestorationforbioremediation.Twobacterialrapidbioassays,SOSchromotestandtheumutestwithandwithoutmetabolicactivation(S-9mixture),wereusedtoevaluatethegenotoxicityofpetroleumhydrocarbon-contaminatedsoilfollowingbioremediationtreatment.ThesoilwastakenfromanengineeredbiopileattheCzechowice-DziedzicePolishoilrefinery(CZOR).Thebioremediationprocessinthebiopilelasted4years,andthetoxicitymeasurementsweredoneafterthistreatment.Carcinogensdetectedinthesoil,polyaromatichydrocarbons(PAHs),werereducedtolowconcentrations(2mg/kgdrywt)bythebioremediationprocess.Genotoxicitywasnotobservedforsoilstestedwithandwithoutmetabolicactivationbyaliverhomogenate(S-9mixture).However,theumutestwasmoresensitivethantheSOSchromotestintheanalysisofpetroleumhydrocarbon-bioremediatedsoil.Analyticalresultsofsoilusedinthebioassaysconfirmedthatthebioremediationprocessreduced81%ofthepetroleumhydrocarbonsincludingPAHs.Weconcludethatthecombinedtestsystemsemployedinthisstudyareusefultoolsforthegenotoxicexaminationofremediatedpetroleumhydrocarbon-contaminatedsoil.r2004ElsevierInc.Allrightsreserved.Keywords:Genotoxicity;Petroleumhydrocarbons;Bioremediation;Biopile;PAHs1.IntroductionThesuccessofbioremediationisdeterminedbythemetabolicpotentialofmicroorganismstodetoxify/utilizeorganiccontaminantsormineralizethemtoCO2,H2O,andbiomass.TheprocessishighlydependentonbiodegradabilityandbioavailabilityofcontaminantsaswellasenvironmentalparametersincludingpH,temperature,andnutrientavailability(Barthe,1986).Bioremediationisanattractive,envir-onmentallyfriendly,andrelativelycost-effectivealter-nativetoconventionalphysicochemicalsoilandwatertreatmenttechniques(Bouweretal.,1994;Barthe,1986).Thereareavarietyofinsituandexsitubioremediationapproaches,includingbiopiling,bio-reactors,bioventing,biosparging,andlandfarmingthatcanbeappliedtooptimizebiodegradation(Brigmonetal.,2002;Hazen,1997;Bouweretal.,1994).Consideringdifferentexistingbioremediationstrate-giesandcharacteristicsofthelagoonmaterialsattheCzechowice-Dziedzicerefinery,theexsitu/onsitebioremediationinbiopilewasselectedtocleanupsoilheavilycontaminatedwithpetroleumwaste(Pazaetal.,2003).Asaresultofbioremediationnosecondarywastestreamiscreated,contaminantsaresafelytreatedinsitu,thereforeminimizinghandlingandassociatedhealthhazards,andthereuseofthepreviouslycontaminatedsitewithoutrestrictionispossible.Anumberofrapidtestsystemstodetectenviron-mentalcarcinogenshavebeendevisedforsoilandwateranalyses.TheumutestandSOSchromotesthavebeenARTICLEINPRESSwww.elsevier.com/locate/ecoenv0147-6513/$-seefrontmatterr2004ElsevierInc.Allrightsreserved.doi:10.1016/j.ecoenv.2004.10.014
Correspondingauthor.Fax:[phone].E-mailaddress:[e-mail](G.Paza).widelyemployedforsoiltestingwithsuccessfulresults(Reifferscheidetal.,1991;McDanielsetal.,1990).TheSOSchromotestdescribedbyQuilardetetal.(1985,1993)usesoneoftheSOSgenes,sfiAfocusedtolacZonthechromosomeofEscherichiacoli,andovercomesthedrawbackintheAmestest.Theumutestisbasedontheuseofgeneticallyengineeredbacteria,SalmonellatyphimuriumTA1535pSK1002.TheumutesthasbeenstandardizedandvalidatedbyGermanDIN(DIN38415T3)andontheinternationallevelbyISO(ISO/DIS13829,2000).Inthisprojectgenotoxicityassess-mentofremediatedhydrocarbon-contaminatedsoilwasperformedwiththeSOSchromotestandumutest.2.Materialsandmethods2.1.FielddescriptionMorethanacenturyofsulfuricacid-basedoilrefiningprocessbytheCZORrefineryandassociatedwastedisposalhasproducedanestimated120,000tonsofacidic,highlyweatheredpetroleumsludgeandwaste-water.Thiswastehasbeendepositedintothreeopenwastelagoons,3mdeepandcovering3.8ha.Oneofthewastelagoons(0.3ha)waschosenforaerobicbiopiledemonstration(Fig.1A)(Pazaetal.,2003).Thewastefromthislagoonwasremoved,andheavilypetroleumcontaminatedsoilwasengineeredintoabiopileforbioremediation.Thebiopilewasconstructedwithactivelyandpassivelyaeratedsectionsin1997(Pazaetal.,2004).Theprojectfocuswasontheevaluationofnovelenvironmentalrestorationtechnologiesandre-searchforeffectivebioremediationofheavilycontami-natedpetroleumwastesoils.Thiswasaccomplishedbycomparingbioremediationprocessesandthecontami-nantremovalratesofbotheasilybiodegradablelightpetroleumhydrocarbonsandmorerecalcitrantpolyaro-matichydrocarbons(PAHs)underactivevs.passiveaeration.Theapplicationefficientlyemployedcost-effectiveadditives,includingmineralNPKfertilizers,thesurfactantRokafenolN8,andemployingindigenousmicrobialconsortium