ABSTRACT

TheIllinoisJuniorAcademyofScience
Thisform/papermaynotbetakenwithoutIJASauthorization.

CATEGORY

EnvironmentalScience

STATEREGION#

SCHOOL

NilesNorthHighSchool

IJASSCHOOL#

6038

CITY/ZIP

Skokie,IL60076

SCHOOLPHONE#

8476262256

SPONSOR

SusiePosnock

MARKONE:

EXPERIMENTALINVESTIGATION

DESIGNINVESTIGATION

NAMEOFSCIENTIST*

ZacharyLuna

GRADE

11

NAMEOFSCIENTIST

GRADE

NAMEOFSCIENTIST

GRADE

NAMEOFSCIENTIST

GRADE

*Ifthisprojectisawardedamonetaryprize,thecheckwillbewritteninthisscientist'sname,anditwillbe
his/herresponsibilitytodistributetheprizemoneyequallyamongallparticipatingscientists.

Luna

PROJECTTITLE

TheEffectofNitrogenandOxygenonMicrobeInducedOilDegradationinLake
Michigan

Purpose:Thepurposeofthislaboratoryexperimentistolearnabouttheconditionsthatwouldcauseoileating
bacterialocaltoLakeMichigantomultiply.Thegreatertheabundanceoftheoileatingbacteriathemoreoil
thebacteriawillbeabletoingest.Sincethesemicrobesareverysensitivetotheenvironmenttheylivein,tests
wouldbeusedtoseeifconditionslikewarmtemperaturesandincreasednitrogencancausetheseoileating
bacteriatomultiply.TestswillbeperformedonoillacedsoilsamplesderivedfromtheMidwestnearLake
Michigan.
Procedure:Thisprocedureinvolvedcalculatingtheoriginaldensityofcontaminatedsoilsamplesascompared
tothesamplesafteroildegradingmicrobeswereintroducedwithoutsidevariables.Variablesintroduced
includeincreasesinnitrogen,oxygenandwarmertemperatures.Thedensityofeachsampleafter7dayswas
thencomparedtotheoriginaldensity.

Conclusion:Overall,thedatasupportedthehypothesis.AnidealvariableofincreasedOxygenandtemperature
wasfoundtobethemostefficientforspeedinguptheprocessofoildegradation.Thisvariablewasproventobe
thedominantamongtheothersamplesbecauseinall3trialsthedensitywasreducedatafasterrateinthe7
dayspanthenanyothervariable.Thus,provingthatthemicrobeswereabletoconsumeandbreakdownoil
fasterbecauseofthesespecificconditions.InlocationssuchastheGulfofMexico,themicrobesfavorthewarm
temperatureandworkbetterwithincreasedNitrogen.However,inanexperimentwithsoilandmicrobesnative
toLakeMichiganwheretheclimateisverydifferentandcanvarydramaticallyasfarastemperature,the
microbesshowadesireforincreasedoxygenwithwarmertemperatures.Oildegradingmicrobescanvaryin
howtheyreacttocertainvariablesbasedontheclimatetheysurvivein.

Luna

1)LimitAbstractto3paragraphs(about200wordsorless).a)Purposewhatyousetouttoinvestigateb)
Procedurehowyoudid
itc)Conclusionbasedonyourresults.Labeleachparagraph.
2)Mustbetyped,singlespacedonthefrontofthisform.Donotwriteonthebackofthisform.
3)ThreecopiesofyourcompletepaperarerequiredattheStateScienceProjectExposition.
FourcopiesofyourcompletepaperarerequiredfortheStatePaperSessionCompetition.

Thisformmustbeused.Thisformmustbedisplayedonthefrontoftheexhibitorsdisplayboard.Itmaybe
reducedtohalfasheetofpaper8.5inches(vertical)X5.5inches(horizontal).

Luna

Luna

TheEffectofNitrogenandOxygenonMicrobe
InducedOilDegradationinLakeMichigan
NilesNorthHighSchool
10/24/14
ZacLuna

Luna

TableofContents:

Acknowledgments................................................................................................................................................3
Purpose.....................................................................................................................................................................4
Hypotheses and Rationales..............................................................................................................................5
Review of Literature.....................................................................................................................................6-13
Materials................................................................................................................................................................14
Procedure.......................................................................................................................................................15-16
Variables................................................................................................................................................................17
Results.............................................................................................................................................................18-21
Data Analysis.................................................................................................................................................22
Experimental Error...........................................................................................................................................23
Conclusion......................................................................................................................................................24
References......................................................................................................................................................25-26

Luna
Acknowledgments
IwouldliketothankMrs.Posnockforkeepingmeontaskandgivingeffective
guidanceasmyprojectprogressed.InadditionIwouldliketothankmymentorswhoare
researchersinmyspecifictopicofstudySteveRockandAnnWhelan.WithoutthemIwould
belackingasignificantamountoftheinformationIhadbeforeevengoingintotesttrials.
Finally,Iwouldliketothankmymomforkeepingmemotivatedandencouragingmetotake
extrameasurestoensuretheprojectwouldbesuccessful.

Luna

Purpose:
Thepurposeofthislaboratoryexperimentistolearnabouttheconditionsthat
wouldcauseoileatingbacterialocaltoLakeMichigantomultiply.Thegreatertheabundance
oftheoileatingbacteriathemoreoilthebacteriawillbeabletoingest.Sincethesemicrobes
areverysensitivetotheenvironmenttheylivein,testswouldbeusedtoseeifconditionslike
warmtemperaturesandincreasednitrogencancausetheseoileatingbacteriatomultiply.
TestswillbeperformedonoillacedsoilsamplesderivedfromtheMidwestnearLake
Michigan.

Luna

Hypothesis:
IfoileatingbacterianativetoLakeMichiganareexposedtoconditionssuchas
warmtemperature,increasednitrogenandincreasedoxygen,theywillmultiplycausingtheoil
inasampletobeingestedatamoreefficientrate.

Rationale:
Thehypothesisisvalidbecausecurrentresearchindicatesthattheseconditions
arefavorableformultiplyingoileatingbacteriacoloniesthathavebeenstudiedinother
terrainsandgeographicregionsofthecountry
.
Bycombiningthesefavorableconditionsitwill
increasethethebacteriasabilityandefficiencyatingestingtheoilinoillacedsoilsamples
fromtheLakeMichiganMidwesternregion.

Luna

ReviewofLiterature
Whenanoilspilloccurs,itendangerstheaffectedplantandanimalspeciesand
disruptstheecosystem.Oilcanpollutedifferenttypesofterrainincludingbodiesofwater,
coastline,andsoil.Althoughoilisanaturaloccurringsubstance,itisstillhazardoustomost
speciesofplantsandanimalsinlargequantities.Largeoilspillscantakeyearstoremove
fromanenvironment.Althoughtherearemanymethodstocleanupoilspills,mostare
inefficientandmaycauseaharmfulimpactontheenvironment.Methodssuchasburning,
skimmingandreleasingchemicalstodepletetheoilhavebeenshowntobeeffectivein
specificenvironmentsbutdonotthoroughlyremovetheoil.
Scientistshavediscoveredthattheearthhaswaystohealitselffromoilspillsand
naturaloilseepagefromtheearthscrust.Specializedmicroorganismshaveevolvedwhich
ingestordegradeoil.Naturallyoccurringmicrobesfeedonhydrocarbonsintheoilwhichthen
breakitdown.Oilcontainshydrocarbonswhichconsistsofdifferentamountofhydrogenand
carbon.Thebacterianeedoxygentocombinewiththehydrocarbonswhichthencreatewater
andcarbondioxide.Theseoileatingbacteriacaneasilybedispersedbyagentssuchas
currents,windsanddeteriorationfromthesoil.Bypropagatingthesenaturallyoccurring
bacterias,orbycreatinglargeamountsofsyntheticoileatingbacteria,amoreefficient
strategymaybefoundtoaddressoilspillswhichmayhavealessharmfulimpactonthe
environment
(Hazen,2013).
Thesespecializedbacteriaarebeingstudiedtolearnhowtheybreakdowntheoiland
underwhatconditionstheythrive.Amyriadoffactorsmustbeconsidered,whichmayaffect
the

Luna
growthoftheseoileatingbacteriacolonies,suchastemperature,nutrients,adhesiontothe
oil,carbon/nitrogenlevels,waterandwatermovement.Certainoileatingbacteriastrains
maythrivebestunderveryspecificconditions.Forexample,oilspillsintheantarcticmay
attractdifferentoileatingbacteriastrainsthanoilspillsintheGulfofMexico.
IthasbeendiscoveredthatbacteriaonthebeachesoftheGulfofMexicoconsumed
theoilfromtheDeepWaterHorizonoilleakbyaddingnitrogentotheirdiet.Astatementwas
madebyaleadresearcherinwhichdifferenteffectivecleanuptechniquesusingtheoilare
explored."Butbecauseoilislowinnutrientssuchasnitrogen,thiscanlimithowfastthe
bacteriagrowandhowquicklytheyareabletobreakdowntheoil,ouranalysisshowedthat
somebacteriaareabletosolvethisproblemthemselvesbygettingtheirownnitrogenfrom
theair(Kostka,2013).Scientiststhenstudiedspecificgenesthatwereresponsiblefor
differentbehaviors,includingnitrogenfixing,inordertofigureouthowthebacteriawere
breakingdowntheoil."Byunderstandinghowtheoilisdegradedbymicrobes,which
microbesdothework,andtheimpactofthesurroundingenvironmentalconditions,wecan
developwaystointervenetosupportthenaturalcleanupprocess(Kostka,2013)."For
instance,maybeincreasingthelevelsofnitrogenforthebacteriatotakeincouldspeedup
thebreakdownoftheoilandthismethodofcleansing.Althoughthismaybeaverybeneficial
catalystforspeedinguptheoilremovalprocess,anykindofhumaninterruptioninthenatural
cleanupshouldbehandledwithcaresoastonotcausemoreunintendeddamagetothe
delicateecosystem.
TheDeepwaterHorizonoilspillpumpednearly5millionbarrelsofoilinatimeframe
of
threemonths.However,withinseveralweeksoftheoilleakbeingfixed,multipleareasofthe
Gulfwerefreeofoil.Aleadresearcherstatedavalidexplanationandconclusionfromthese

10

Luna
results.ThebottomlinefromthisresearchmaybethattheGulfofMexicoismoreresilient
andbetterabletorecoverfromoilspillsthananyonethought.Itshowsthatwemaynotneed
thekindsofheroicmeasuresproposedaftertheDeepwaterHorizonspill,likeaddingnutrients
tospeedupthegrowthofbacteriathatbreakdownoil,orusinggeneticallyengineered
bacteria.TheGulfhasabroadbaseofnaturalbacteria,andtheyrespondtothepresenceof
oilbymultiplyingquiterapidly(Hazen,2013)."ThereasontheGulfissoresilienttooilspillsis
becauseeachyearasmanyas1.4millionbarrelsofoilenterthegulffromnaturalseepsin
theearthscrust.Overtheyearsevolutionhascreatedahugepopulationofoileatingbacteria
intheGulf(Koebler,2013).AlthoughtheGulfmaybebetteroffthaninitiallythought,notall
ecosystemsarelikethisandsomestillneedhumaninfluencesforthebacteriatoreallykeep
uptospeedagainstthedetrimentaleffectsofoil.However,Insomelocations,ittookonly
onedayforthebacteriatoreduceagallonofoiltoahalfgallon(Hazen,2013).
AlthoughTheDeepWaterHorizonoilspillwaseasilyassistedbythehelpof
microbes,manyotherspillslackthisgiftofnaturalcleansingduetotheenvironment.
Recently,theLakeMichiganregionhasexperiencedtroublewithoilspills.BPrecentlymade
afalsestatementabouttheamountofoilspilledinthelake.
Inastatement,the
company
saidamalfunctioninanewdistillationunitforcedupto39barrelsor1,638gallonsofoilinto
thelakejustacrosstheIllinoisborder.Adayearlier,thecompanyhadestimatedthat18
barrelsatmosthadbeenspilled.(Hawthorne,2014).
Oilcompaniesoftenintentionallyfib
abouttheamountofoilspilledduetothecleanupbeingsocostly.BPiscurrentlyusing
vacuumtrucksandothermechanicalcleansingdevicesinordertokeeptheoilclearfromthe
drinkingwaterofthe7millionpeoplelivinginChicago.Althoughthisisagoodtemporary
methodofcleansing,itisnotveryefficientandoftenleavesresidueoilbehind.Evenafter
cleanup,oilstillremainsembeddedinshorelines,rocksandsoil.

Crewsdeterminedthattrace

11

Luna
amountsofoilremainalongtherockyshorelineofBP'srefineryinWhitingfollowingtheMarch
24oilspill.Upto1,638gallonsofoilweredischargedwhenadistillationunitmalfunctioned
(Jackson,2014).Thisclearlystatestheoilcleansingmethodsoftodayarenotefficiently
cleaningspills.Machineryandothermethodsoftentakealongspanoftimeandarenot
thorough.Oilspillsnearlargecitiesneedtobecleansedasquicklyaspossiblesoastonot
affectthedrinkingwater.
Inanevenmorerecentincident,theOlympicOilspillresultedinthousandsofgallons
ofoillostintoLakeMichiganandthesurroundingarea.Thecontaminationhasnowmigrated
toaChicagoareacanalandtheEPAhasgotteninvolvedwithacourtorderrequiringthe
Cicerocompanytocleanupthemess.The
EPA

saidTuesdaythatitsaskedAttorney
GeneralLisaMadigans
office
toobtainanorderrequiringOlympicOilLtd.,tostopthespill,
determineitssizeandcleanupcontaminatedsoilandwater.Thefacilityblendsandpackages
petroleum
lubricatingoilandantifreezeproducts(Madigan,2014).Sincethegeographyandclimate
surroundingtheLakeMichiganareadiffersgreatlyfromtheGulfofMexicoregionwherethe
DeepWaterHorizonspilloccurred,ittwouldbeveryhelpfultolearnaboutthecharacteristics
oftheoileatingmicrobesnativetothisspecificenvironment.Sincetheselifeformsarevery
sensitivetoconditionsintheirecosystem,propertiesthatwouldspeeduptheefficiencyofoil
consumptionmayvaryduetothedifferenceintheecosystem.Ifafavorablemethodisfound
fortheenvironmentofLakeMichigan,spillssuchasthesecanbemoreeasilycleanedina
naturalwaysthatdonotfurthernegativelyimpacttheenvironmentorharmthedrinkingwater
supplygoingtotheChicagolandarea.
Someoftheoilspillresearchhasfocusedonfertilizinghydrocarbonoxidisingbacteria
sotheycanbetterbreaktheoildownintohydrogenandcarbondioxide.Thisstrategyis

12

Luna
effectiveinmarshlikewatersthatarefairlystagnantanddonothavemuchofacurrent.
Howeveramain
limitationisthecontinuousdilutionofthebacteriainopenandmovingwaters.Anewmethod
wasdevelopedwhichinvolvedaddingthinmineralfilm,suchascertaintypesofearthclay,
withthebacteriatotheoilspill.Bydoingthis,thebacteriacanbetteradheretotheoilandnot
driftoffanddisperseinmovingwaters.TogetherwithadsorbedNandPfertilizers,filming
additives,andorganoclay,clayflakescanbeengineeredtofloatonseawater,attachtothe
oil,andslowlyreleasecontainednutrients.Ourlaboratoryexperimentsofmicrobialactivityon
weatheredsourceoilfromtheDeepwaterHorizonspillintheGulfofMexicoshowfertilized
claytreatmentsignificantlyenhancedbacterialrespirationandconsumptionofalkanes
comparedtountreated
oilinwaterconditionsandreactedfasterthanstraightfertilization(Warr,2013).

Ifthis
fertilizerlikesubstanceisaddeddirectlytotheoilspill,thenthiscouldeasilyreducethe
durationoflargebodiedwateroilspills.However,beforethiscanbeappliedtoopenwaters,
aneffectivemethodmustbefoundforkeepingthesupplementsstucktotheoilforlong
durationsoftimesothebacteriacangrowlocally.
Anotherwaytoincreasetheeffectivenessofoileatingbacteriaisbyboostingthe
exportsoftheenzymethatmultipliesthebacteria."WhatwedoisaddFyreZymetoexisting
bacteriaineffect,forcefeedingthemsotheymultiplyandconsumethecontaminationata
muchmorerapidpace.Asaresult,wecandointhreetosevenweekswhatwouldtake
natureanywherefrom30to70yearstocomplete(Condy,2013)."Thiswouldbehelpfulin
waterssuch
astheantarcticoceanbecauseduetothecoldtemperaturesthebacteriamovemuchslower
thantheywouldinawarmplacesuchastheGulfofMexico.TheFyreZymeispumpedinto

13

Luna
thegroundwithoxygenandwatersothattheFyrezymecanefficientlygettothe
contaminatedareaofsoil.Althoughthereexistothermethodstodecontaminateoildrenched
soil,mostworkmuchslowerandhaveaftereffects
.
Theseincludevaporextraction,where
holesarepunchedintotheground,airispumpedin,andcontaminantsareextractedthrough
acarbonfilter.Whileeffective,thisprocesscantakefrom7to15years.Anothercommon
treatmentwidelyutilizedhasbeen"thermaldesorption,"aprocedurewherethesoilisburned
toeliminatethecontaminants.However,thisprocessalsosterilizesthesoil,renderingit
uselessforsustainingplantlife(Cultice,2013).
Bacteriahasalsoshowntobeabletomovethroughsoilandcoastlines.Howthe
bacteriacandothisandseekouttheoilisstillunknown.Thisiswhydealingwithoilonlandis
nowakeyareaofresearch.Howtheoildispersantsandbacteriamovethroughporous
materiallikesandandmudwouldbeahugehelpforcleaninguponshoreoilcontaminations.
Oncetheoilreachessedimentsandbeachesandbayous,howdoesittransportitself
throughtheporousmediumthatthosebeachesandbayousconstitute.Buttheothervery
importantquestionthatwe
cananswerthroughthisishowwecaninterveneinorderfortheoiltoberemoved
(Papadopoulos,2011).Onlandoilspillshavebeenahotspotforcleanupmethodslike
burningandskimmingthatareharmfultotheenvironment.Byfiguringouthowthebacteria
getaround
andhowhumanscanintervenetoincreasethisfunction,wouldhelpthecausetremendously.
Differentenvironmentscallfordifferentcleanupmethodssoasnotto
disturbtheland.
We
arelookingatallthesefactorstooptimizethewindowofreagentsavailablefortakingcareof
oilspills.Whywindow?BecausedependingonwherethespillisintheGulf,orinAlaska,
thetemperatureisdifferent,thesalinityisdifferent(Somasundaran,2011).

14

Luna
Testingcontaminatedsoilusingdifferentstrainsofoileatingbacteriawouldallowusto
seewhatconditionscausethebacteriatothriveandremovetheoilfromtheenvironment.
Puttingthedifferentstrainsunderavarietyofphysicalconditions,andtestingwithdifferent
soilsamples,willgiveawidevarietyofenvironmentalscenariosthatwouldyielddataonhow
bestthesebacteriascanthriveinourGreatLakesMidwesternenvironment.Collecting
samplesfromrealcontaminatedoilsites,andcreatingcontaminations,willstretchallpossible
areasofresearch.Theexperimentseekstofindsignificantchangesinsoilsamplegrease
testsunderdifferentconditionsafterspecifiedtimespans.
Overall,enlistingmicroorganismstoassistwiththecleaningupofoilspillsisthemost
ecofriendlywaytogo.Itisaprocessalreadyoccurringinnatureandispartoftheearths
naturalhealingprocess.Howeverfactorssuchas
temperature,nutrients,adhesiontotheoil,
carbon/nitrogenlevels,waterandwatermovementplayahugeroleinthisoileatingbacteria
cleansingmethod.Thesefactorsmustbetakenintoaccountandalteringsomeofthese
factors
couldspeeduphowfastthebacteriadoesitsjob.Ifhumansaretointerferewiththisnatural
cleansing,thenitshouldbehandledwithgreatcautionsonounintentionalharmiscausedto
theenvironment.
Differentformsofcontaminationfromoilisalsoahugeresearcharea.Soil
decontaminationwouldofcourserequiredifferentmethodsthanwater.Findingouthowthe
bacteriaworkincleaningthesoilwouldbeahugehelp.Thiscouldleadtofiguringouthow
humanscaninterveneandincreasethebacteriaseffectiveness.Byaddingnitrogenoroxygen
andincreasingthetemperatureforthebacteria,thismayspeeduptheprocessofbreaking
downtheoilandcauselessenvironmentaldamage.Thishasbeenfoundtobethecasein
regionsotherthantheMidwestasdescribedabove.

15

Luna
InconclusionthemicrobeswhichwillbetestedarenativetotheLakeMichiganregion.
Therefore,thehypothesisthat
ifoileatingbacterianativetoLakeMichiganareexposedto
conditionssuchaswarmtemperature,increasednitrogenandincreasedoxygen,theywill
multiplycausingtheoilinasampletobeingestedatamoreefficientrate
isapredictionone
mightnaturallyconcludefromthecurrentresearch.Itisalsorelevant,worthwhileresearch
duetoproblemswithoilspillswhichhaveoccurredinrecentyearsinthisregion.TheGreat
Lakesregionisanareawhichprovidesdrinkingwatertoalargepopulationofpeople.
Understandingandstudyingtheoileatingbacteriainthesoil(andalsothewater)intheLake
Michiganregioncouldhavegreatbenefitstoaddresscurrentoilspillsinthisregionandoil
spillsthatmayoccurinthefuture.

Materials:
3gramsNitrogenfertilizer(Urea)
Oxygenstones(1percontainer)
Oildegradingmicrobes(3grams)
Samplescollectedfromlocalsites(LakeMichiganshore)
Scale
Graduatedcylinder
Scoopula
Isolatedandcontainedareaformicrobestoculture
Incubator
Individualcontainersforsamplesbeingtested.

16

Luna

Procedure:
1. Acquiresamplesofoillacedsoilfromlegitimateoilspills.Thesespillsmustbenative
toLakeMichiganbecausethatiswhattheexperimentwillbespecifiedas.
2. Placeeachsampleinsideofgraduatedcylinderandmeasureoutthevolumein
milliliters.
3. Weigheachsamplewithscaletofindmassingrams.
4. Calculateoriginaldensityofcontaminatedsoilsamples.
5. After3gramsofthemicrobeshavebeenintroducedtothesamplesortheyare
alreadypresent,theymustbeplacedincontainedareasotheycancontinueto
cultivateandvariablescannowbeadded.
6. Fortrialone,therewillbefourdifferentvariablesandonecontrolwherejustmicrobes
arepresentinthesample.

17

Luna
7. Eachvariablewillbeanalyzed2timestoseeprogressinoildegradation.Therewill
beasecondtrialandthirdofnewsamplesandtheprocesswillthenwerepeated.
8. Thefirstsamplewillbeusedasthecontrolwherenovariableswillbeintroduced.
9. Thesecondsamplewillhavethevariablepresenceof3gramsNitrogenintroducedto
themicrobes.
10. ThethirdsamplewillhavethevariablepresenceofincreasedNitrogenandincreased
temperaturetothemicrobes.
11. ThefourthsamplewillhavethevariablepresenceofincreasedOxygentothe
microbes.
12. ThefifthsamplewillhavethevariablesofincreasedOxygenlevelsandincreased
temperature.
13. IncreasednitrogencanbeaddedwithUrea(3grams)
14. Increasedoxygencanbeaddedbyintroducingwatertoanoxygenstone
15. Temperaturemustbeincreasedinanincubator(32.6Celsius).
16. Oncemicrobesandvariableshavebeenapplied,allowmicrobestobeginoil
degradation.
17. Thedensityofeachsamplewillthenbecalculatedtwicebydividingthemasstothe
volume.
18. Calculatedensityofeachsample3daysaftermicrobeshavebeenapplied.
19. Calculatedensityofeachsample7daysaftermicrobeshavebeenapplied.
20. Thesestepswillthenberepeatedfortrials2and3.
21. Alldatashouldbedocumentedwithdetailforeachtrial.
22. Chartalldataandcomparenewdensitytooriginalthatwascollectedbeforethe
microbesandvariableswereapplied.

18

Luna

Variables:
The
independentvariable
isthephysicalfactorsthatwillbeintroducedtotheoileating
bacteriasuchashigherlevelsofNitrogen,Oxygenandwarmertemperatures.
The
dependentvariable
ishowefficientthebacteriacleansedthecontaminatedsoildueto
theintroducedphysicalfactors.Thiswillbemeasuredbycalculatingthenewdensityand
comparingittotheoriginalofthesamplesoncethetestshavebeenconducted.
The
control
groupisthemicrobesaloneintroducedtoasamplewithnooutsidevariables
involved.

19

Luna

Trial1:

Days:

Nitrogenincrease

1.677g/ml

1.648g/ml

1.339g/ml

20

Luna

Nitrogenandtemp
increase

1.477g/ml

1.405g/ml

1.122g/ml

Oxygenincrease

1.447g/ml

1.438g/ml

1.217g/ml

Oxygenandtemp
increase

1.9765g/ml

1.893g/ml

1.663g/ml

Control

1.682g/ml

1.672g/ml

1.574g/ml

Days:

Nitrogenincrease

1.8085g/ml

1.745g/ml

1.611g/ml

Trial2:

21

Luna

Nitrogenandtemp
increase

1.643g/ml

1.5225g/ml

1.1684g/ml

Oxygenincrease

1.5425g/ml

1.4475g/ml

1.3725g/ml

Oxygenandtemp
increase

1.9625g/ml

1.8485g/ml

1.3455g/ml

Control

1.546g/ml

1.537g/ml

1.4297g/ml

Days:

Nitrogenincrease

2.2205g/ml

2.138g/ml

1.6945g/ml

Trial3:

22

Luna

Nitrogenandtemp
increase

1.5775g/ml

1.537g/ml

1.4297g/ml

Oxygenincrease

2.086g/ml

2.008g/ml

1.7265g/ml

Oxygenandtemp
increase

2.241g/ml

2.112g/ml

1.4715g/ml

Control

2.2835g/ml

2.2445g/ml

2.0885g/ml

AverageDensityGraph:

23

Luna

DataAnalysis:

24

Luna
Aweekwasgiveninordertofindsignificantresultsinthedecreaseofdensityperoillacedsoil
sample.Thiswasshowntobesuccessfulbecausebythe7thdaysomesampleswereabletodecreasein
suchasubstantialamountthatitwasvisiblewhenactuallyviewingthesamplethatsomeoftheoilhadmost
definitelybeenconsumedbythemicrobes.Althoughallsamplesshowedadecreaseindensity,thecontrol
withmicrobesalonehadamuchlowerdecreaseinthecourseofthe7days.Thisfurtherbacksupthefact
thatoildegradingmicrobescanbemanipulatedbyoutsidevariables.
Thedataprovedtosupportthehypothesisandgainsuccessfulresultsbecausethemicrobeswere
abletoincreasetheirrateofoilconsumptionbybeingintroducedtonumerousdifferentvariablessuchas
increaseinOxygen,Nitrogenandtemperature.Thisresultedinthemicrobesdramaticallyaffectingthe
densityofthesoilinaslittleas7days.Basedonthedatacollected,theaveragedensityof7daysforthe
Nitrogenandtemperatureincreasesampleswas1.18975g/ml.Sincethiswasthelowestdensityaverage
comparedtothecontrolaverageof1.6974g/ml,itisportrayedasthemostefficientvariablegroupforoil
consumption.However,whenallaveragedensitiesforeachrecordeddaywerecalculatedandplacedina
graph,itshowedthattheOxygenandtemperatureincreasewasactuallythemosteffectivevariablegroup
forthemicrobesinthe7dayspanoftime.Themicrobesareshowntohaveconsumedthemostoiloverall
inthisgroupbecauseofits.43g/mldensitydifferencefromthefirstdaytothe7th.Thecontrolactuallyhad
thelowestdensitydecreasecomparedtotheother4variables.

ExperimentalError:

25

Luna
Thenumberoneexperimentalerrorintheprojectwouldmostlikelybethelossofsoilwhen
calculatingthedensity.Thesoilmustbeweighedandplacedinaseparatecontainerforvolume.Thesoilis
veryadhesiveandnoteverysinglegrainofsoilcanbeputbackintotheoriginalsamplecontainer.Thiscan
slightlyaffecttheoverallresultsbutnotonalargeenoughscalefortheexperimenttobeinaccurate.Another
errornoticedinthetestingprocesswastheoxygenstonesandthefactthattheyneededwaterinorderto
giveofoxygen.Oncethewaterwasappliedtothestonesatthebottomofthesamples,thedensitywas
mostlikelyslightlyaffectedsincethewaterwasabsorbedbythesoileventually.Anothererrorwouldbe
addingincorrectmeasurementsofthevariablesaddedsothattheywerenotkeptconstantinalltrialsand
samples.Thiscouldhavehappenedwhenweighingthenitrogenfertilizeronthescaleortheamountof
wateraddedtoeachoxygenstone.

Conclusion:

26

Luna
Overall,thedatasupportedthehypothesis.AnidealvariableofincreasedOxygenandtemperature
wasfoundtobethemostefficientforspeedinguptheprocessofoildegradation.Thisvariablewasproven
tobethedominantamongtheothersamplesbecauseinall3trialsthedensitywasreducedatafasterrate
inthe7dayspanthenanyothervariable.Thus,provingthatthemicrobeswereabletoconsumeandbreak
downoilfasterbecauseofthesespecificconditions.InlocationssuchastheGulfofMexico,themicrobes
favorthewarmtemperatureandworkbetterwithincreasedNitrogen.However,inanexperimentwithsoil
andmicrobesnativetoLakeMichiganwheretheclimateisverydifferentandcanvarydramaticallyasfaras
temperature,themicrobesshowadesireforincreasedoxygenwithwarmertemperatures.Oildegrading
microbescanvaryinhowtheyreacttocertainvariablesbasedontheclimatetheysurvivein.
Thistopicandprojectcanhelpimprovethecleanupofmanmadeoilspillsinanatural,nonharmful
way.Bylearningmoreaboutthesespecialmicrobes,societycanfindtheidealvariablesbasedonthe
climateforthebacteriatoworkatfullpotential.Anoilspillwhichwasonceaverydifficult,longand
damagingprocesstocleanup,cannowbeanissueofthepast.Manipulatingthisnaturalformofcleanup
canhelphumansfixmessyoilmistakeswithoutruiningtheearthintheprocess.

ReferenceList

27

Luna
BiotechnologyResearchInternational.(2013,May27).BioremediatingOilSpillsinNutrientPoorOceanWatersUsing
FertilizedClayMineralFlakes:SomeExperimentalConstraints.RetrievedDecember13,2013,from
http://www.hindawi.com/journals/btri/2013/704806/
Cultice,C.K.(n.d.).FyreZyme.In
Let'sdolunchCommerceDepartmentgranthelpsfightpollutionbyboostingexportsof
enzymethatmultiplesoileatingbacteria.
. RetrievedDecember14,2013,fromEBSCOHOST(510390439).
Faeber,F.(2011,April5).OilEatingBacteriaEngineering.In
NationalGeographic
.RetrievedDecember14,2013,from
http://news.nationalgeographic.com/news/2011/04/110405nsfoilsomasundaranvideo/

Hawthorne,Michael."BPraisesestimateofLakeMichiganoilspill."
ChicagoTribune
.N.p.,28Mar.2014.Web.1Nov.2014.
<
http://articles.chicagotribune.com/20140328/news/chibpoilspilllakemichigan20140327_1_londonbasedoilcompanyoil
spillwhiting
Jackson,Angie."CrewsfinishcleanupafterBPoilspillonLakeMichigan,traceamountsofoilremain."
MichiganLive
.N.p.,4
Apr.2014.Web.8Nov.2014.
<http://www.mlive.com/news/grandrapids/index.ssf/2014/04/crews_finish_cleanup_of_bp_oil.html>.
Kemsley,T.(2013,August30).BacteriaNaturally'EatUp'OilSpillContaminationbySupplementingDietwithNitrogen.
In
NatureWorldNews
.RetrievedDecember13,2013,from
http://www.natureworldnews.com/articles/3757/20130830/bacterianaturallyeatupoilspillcontaminationsupplementingdietn
itrogen.htm
KubeM,ChernikovaTN,AlRamahiY,BeloquiA,LopezCortezN,GuazzaroniME,HeipieperHJ,KlagesS,Kotsyurbenko
OR,LangerI,NechitayloTY,LnsdorfH,FernndezM,JurezS,CiordiaS,SingerA,KaganO,EgorovaO,PetitPA,Stogios
P,KimY,TchigvintsevA,FlickR,DenaroR,GenoveseM,AlbarJP,RevaON,MartnezGomarizM,TranH,FerrerM,
SavchenkoA,YakuninAF,YakimovMM,GolyshinaOV,ReinhardtR,&GolyshinPN(2013).Genomesequenceand
functionalgenomicanalysisoftheoildegradingbacteriumOleispiraantarctica.
Naturecommunications,4
PMID:
23877221
NaetherDJ,SlawtschewS,StasikS,EngelM,OlzogM,WickLY,TimmisKN,&HeipieperHJ(2013).Adaptationofthe
hydrocarbonoclasticbacteriumAlcanivoraxborkumensisSK2toalkanesandtoxicorganiccompounds:aphysiologicaland
transcriptomicapproach.
Appliedandenvironmentalmicrobiology,79
(14),428293PMID:
23645199
OileatingBacteria.(n.d.).In
BiotechnologyOnline
.RetrievedDecember13,2013,from
http://www.biotechnologyonline.gov.au/pdf/enviro/oileating_bacteria.pdf
Schwartz,A.(2013,April8).OilEatingBacteriaFixedTheDeepwaterHorizonDisaster,ButTheyMayNotHelpNextTime.In
Co.Exist
.RetrievedDecember13,2013,from

28

Luna
http://www.fastcoexist.com/1681764/oileatingbacteriafixedthedeepwaterhorizondisasterbuttheymaynothelpnexttime
Shelton,LeeAnn."Cicerocompanyorderedtocleanupoilspillsnearcanal."
Newsfeed
.N.p.,10Oct.2014.Web.6Nov.2014.
<
http://chicagosuntimes.com/news/cicerocompanyorderedtocleanupoilspillsnearcanal/
>.

29