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ABSTRACT

TheIllinoisJuniorAcademyofScience
CATEGORY

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Microbiology
6
STATEREGION#

SCHOOL

NilesNorthHighSchool

IJASSCHOOL#

6038

CITY/ZIP
SPONSOR

Skokie,IL60076

SCHOOLPHONE#

8476262256

SusiePosnock

MARKONE: EXPERIMENTALINVESTIGATION

DESIGNINVESTIGATION

NAMEOFSCIENTIST*

MirnesNeziric

GRADE

NAMEOFSCIENTIST

GRADE

NAMEOFSCIENTIST

GRADE

NAMEOFSCIENTIST

GRADE

*Ifthisprojectisawardedamonetaryprize,thecheckwillbewritteninthisscientist'sname,anditwillbehis/herresponsibilitytodistribute
theprizemoneyequallyamongallparticipatingscientists.

PROJECTTITLE

PossibilitiesofusingMetallicNanoparticlesasAntimicrobialagentsinWrestlingmats

Purpose:Wrestlersareexposedtomanypathologicalbacteriaonawrestling,andmanyoftenbecomesick,withthemajority
ofdiseasescausedbyStaphylococcusaureusandStreptococcuspyogenes.Thisexperimentwasdesignedtoexplorethe
possibilityandcosteffectivenessofusingsilverand/orcoppernanoparticlesasantibioticagentswithinwrestlingmats.
Procedure:BothStaph.aureusandStrep.pyogeneswereculturedinthreedifferentconditions,onfivepiecesofwrestling
matseachacontrol(nonanoparticles),silvernanoinfusedmats,andcoppernanoinfusedmats.Thenanoparticleswere
introducedtothematsbystirringthematsinananoparticlesolutionat400RPMfor24hours.Theconcentrationofthe
solutionwas10ppm.ThematswerelaterobservedunderaSEMatvaryingmagnifications,thencomparedtoeachotherand
thecontroltodeterminetheeffectivenessandcosteffectivenessofthenanoparticles.

Conclusion:Thehypothesiswasneithersupportednorrefuted.Theoriginalpurposeoftheexperimentwastoevaluatethe
effectivenessofsilverandcoppernanoparticlesasantibioticagentswithinwrestlingmats.However,thebacterialcultures
wouldnotgrowonanyofthewrestlingmats,includingthecontrol.Therefore,withnothingtocomparethenanoparticle
treatedmatsto,theeffectivenessofthesilverandcoppercouldnotbedetermined.Possiblereasonsforthelackofgrowthwas
thefactthatthebacteriaused,whileofthesamegenus,werenotthetypicalpathogenicbacteriacommonlyfoundonwrestling
mats.Itisalsopossiblethatpriorantibiotictreatmentofthematspreventedanybacterialgrowthfromstarting.

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


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PossibilitiesofusingMetallic
NanoparticlesasAntimicrobialagents
inWrestlingmats


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TableofContents
TitlePage1
TableofContents2
Acknowledgments...3
PurposeandHypothesis..4
BackgroundInformation..56
MaterialsandMethodofProcedure...711
Variables...12
Data/Results1314
Conclusion....15
References...1617


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Acknowledgments
Ithankthefollowingpeopleforhelpingmyendeavors:
Ms.Posnock
,forhelpingmebrainstormideas,keepingmefromprocrastinating,
andaccommodatingformyotherobligations
Dr.Weiss
,forteachingmehowtoproperlydisposeofhazardouslabwaste
Mr.Thielsen
,forlettingmeborrowhisDremelsaw
ZarahDominandAmbriaBenesch
,formentoringmeduringmyfirstyearof
SIRS,aswellasteachingmetousetheSEM
BillyBertandLarryLau
,forhelpingmeputmyboardtogether


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PurposeandHypothesis

Purpose:
Todeterminetheeffectivenessandcosteffectivenessofsilverand/orcopper
nanoparticlesasantibioticagentsinwrestlingmats.
Hypothesis:
Ifthewrestlingmatshavethesilverorcoppernanoparticlesintegrated,therewill
belittleornogrowthofbacteriaonthem.
Rationale:
Silvernanoparticleshavebeenproventokillbacteriabyinterferingwiththeircell
walls,andbothsilverandcopper,intheirmacroscopicforms,havelongbeenknowntohave
antibioticproperties.
AreaofStudy:
Microbiology,MaterialsScience


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BackgroundResearch

MaterialEngineering,asitisknowtoday,isanewdiscipline,buthasbeenpresentinthe
pastindifferentforms.WhenancientblacksmithsfirstcreatedBronzebycombiningcopperand
tinin4500b.c.e.,theywereusingmaterialengineering.MaterialEngineeringwasoftentermed
metallurgy,duetothefocusoncraftingmetalalloys.Before1955,mostUniversitieshada
Metallurgydepartment,and,toasmallerextent,aCeramicsdepartment.Inthepostwarperiod,
therehadbeengreatadvancesinthefieldofPolymerengineering.Ratherthanhaveaseparate
Metallurgy,Ceramics,andPolymersdepartment,NorthwesternUniversitycreatedtheMaterials
Engineeringdepartmentin1060.
Nanotechnologyisanewfieldofmaterialsengineering,wherescientistsworkwith
extremelysmallparticles,assmallas1nminsizetoas(relatively)largeas100nm.Atthis
scale,materialstendtoexhibitspecialproperties,duetothehugesurfacearea,relativetothe
massoftheatom.Nanotechnologyfirstimaginedin1959byrenownedphysicistRichard
Feynman.Hediscussedthepossibilityofcreatingnewmaterialsbydirectlymanipulatingatoms.
Nanotechnologygainedtractioninthe1980s,withtwoimportantbreakthroughs.In1981,the
scanningtunnelingmicroscopewasinvented,allowingscientistsandengineerstoseeatomsand
theirbonds,andwasusedtosuccessfullymanipulateatomsin1989.In1985,thediscoveryof


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Fullerenes,threedimensionalcarbonmolecules,openedthedoorfortheuseofnanotechnology
inelectronicsandotherpracticalapplications.In2003,therewasthefirstcommercialuseof
nanotechnologyinanantimicrobialrole,withSamsungsSilverNanotechnologybeing
incorporatedintoalloftheirappliances,withtheaimofinhibitingbacterialgrowthonthose
appliances.
Silverhasbeenusedthroughouthistoryasaneffectivetreatmentagainstbacteria.
Hippocratesdiscussedtheuseofsilverinwoundcare,andbythe20thcentury,surgeonshad
beguntousesilversuturestoreducetheriskofinfection.Silverhaslongalsobeenusedasaway
tokeepbacteriafromgrowingandspoilingfoodandwater.ThePhoeniciansstoredwaterand
wineinsilvercoatedvesselsinordertopreventtheliquidfromspoilingduetobacteria.Water
storedonshipsandplanesisoftensilvered,whereasilvercompoundisdispersedintothe
water.Silverwareisalsothebestexampleoftheoligodynamiceffect,atoxiceffectofheavy
metalionsonmicroscopiclivingthings.Silvercutleryanddoorknobs,normallymajorsourcesof
bacteria,willselfsanitizeinafewhours.


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MaterialsandMethodofProcedure

Materials:
1LBeakerx2

Distilledwaterx2L

Nanosilverpowderx~.01g

Nanocopperpowderx~.01g

Wrestlingmatpiecesx30,2.5x3.8cm

40cmmagneticstirrerx2

Papertowels


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Parafilm

Magneticstirplatex1

Razorbladeorexactoknifex1

ElectronScanningMicroscopex1

StaphylococcusepidermidisMicroKwikCulturevialx1

StreptococcuspyogenesMicroKwikCulturevialx1

PreparationofBacteria:
1. Removetheculturevialfromtheresealablebag,andwritetheculturenameonthevial
andonthetesttubeoftherehydrationmedium

2. Removethemetalbandandgraybutylstopperfromthevial

3. Usingapipette,remove1mlofrehydrationmediumandaddittothecultureinthevial

4. Mixgentlywiththesterilepipette

5. Transferthedehydratedculturebacktothetubeofrehydrationmedium


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6. Incubatetheculturefor2448hoursat37C

Experimentation:
1. Cut10pieces2.5x3.8cmofwrestlingmatandplacefiveeachintwopetridishes,
labelingthem
Streptococcus
controland
Staphylococcus
control

2. Createasilvernanoparticlesuspensionbycombining~.01gofnanosilverwith1Lof
waterina1Lbeaker

3. Placeamagneticstirrerinsidethebeaker

4. Placebeakeronpapertowels

5. Cut5x2.5x3.8cmsquaresofwrestlingmatfoam

6. Addthe5wrestlingmatcubestobeaker

7. Submergewrestlingmatcubesinsuspension

8. Sealwithparafilm

9. Placebeakeronmagneticstirplate

10. Settostirat~400rpm:cubesshouldmoveinavortexlikemotion


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10

11. Allowcubestostirinsuspensionovernight

12. Turnoffstirplate

13. Unsealbeaker

14. Removethecubes

15. Setasideinadrydarkplaceandallowtodry

16. Repeatsteps414twicemore,foratotalof15piecesofwrestlingmat

17. Separatethepiecesequallyintothreepetridishes,labeled
Streptococcus
Silver,

Staphylococcus
SilverandSilver

18. Repeatsteps115,butnowwithnanocoppersolution(alterpetridishlabelsappropriately)

19. Apply20lofthe
Streptococcus
rehydrationmediumontocubeswithina
Streptococcus
petridishwithinapetridish,applytofoamsideof3piecesandvinylsideof2pieces

20. Repeatstep19for
Staphylococcusaureus

21. Letculturesgrowonmatsovernight

22. Observethematsunderascanningelectronmicroscope,andrecordthebacteria
populations,andcomparetothecontrolgroup


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Disposal:
1. Placethepetridishesinanautoclave,andsterilizewithheat.

2. Placeallleftovernanoparticlesolutioninanorangechemicalwastecontainer.Clearly
labelthecontainerwithyourname,thenameofthechemical,andandthedateitwas
placedintothecontainer


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Variables
IndependentVariable:WrestlingMatcubesweresubjectedtotwotreatments:1)
Solutioncontainingsilvernanoparticlesat10ppm,and2)Solutioncontaining
coppernanoparticlesat10ppm

DependantVariable:SEMwillcaptureimagesofwrestlingmatatvarying
magnificationstodetermineaveragepopulationofbacterialcolonies.

Control:WrestlingMatsnottreatedwithanynanoparticles

ControlledVariables:Proceduresfollowed,cellenvironment,equipmentused,
equipmentlocation,andsoftwareusedfordataanalysis.


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Data
SEMPhotographs
Description

Magnificationlevel

Foamside,infusedwith
silvernanoparticles

x300

Photo

Foamside,infusedwith
silvernanoparticles

x1000


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Foamside,infusedwith
silvernanoparticles

x3000

Foamside,infusedwith
coppernanoparticles

x300

Foamside,infusedwith
coppernanoparticles

x1000

Foamside,infusedwith
coppernanoparticles

x3000

Analysis


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Thenanoparticleshavebeensuccessfullyintegratedintothematsinclusters.
Therehavebeennosignsofbacterialgrowth.

Conclusion
Thehypothesiswasneithersupportednorrefuted.Theoriginalpurposeofthe
experimentwastoevaluatetheeffectivenessofsilverandcoppernanoparticlesas
antibioticagentswithinwrestlingmats.However,thebacterialcultureswouldnot
growonanyofthewrestlingmats,includingthecontrol.Therefore,withnothing
tocomparethenanoparticletreatedmatsto,theeffectivenessofthesilverand
coppercouldnotbedetermined.Possiblereasonsforthelackofgrowthwasthe
factthatthebacteriaused,whileofthesamegenus,werenotthetypical
pathogenicbacteriacommonlyfoundonwrestlingmats.Itisalsopossiblethat
priorantibiotictreatmentofthematspreventedanybacterialgrowthfrom
starting.


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16

References
Backgroundresearch:

Alexander,J.W.(2009,June).HistoryoftheMedicalUseofSilver.In
PubMed
.Retrievedfrom
Pubmed(19566416).

CommonWrestlingSkinDiseases.(n.d.).RetrievedDecember5,2014,from
http://wrestling.isport.com/wrestlingguides/commonwrestlingskindiseases

Dillow,Clay."StudyFindsCommonlyUsedSilverNanoparticlesAreDeadlytoMicrobes,Plants."
PopularScience
.QueensUniversity,Apr.2011.Web.18Sept.2014.

Fellman,M.(2005,October18).World'sFirstMaterialsScienceandEngineeringDepartmentTurns
50:NorthwesternUniversity.RetrievedDecember4,2014,from
http://www.northwestern.edu/newscenter/stories/2005/10/materials.html

Hwang,Insok,JiHongHwang,HyeminChoi,KeukJunKim,andDongGunLee."Synergistic
effectsbetweensilvernanoparticlesandantibioticsandthemechanismsinvolved."
Journalof
MedicalMicrobiology
.KyungpookNationalUniversity,5Sept.2012.Web.13Oct.2014.


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Jablowicz,M.(2014,March).IntegratingSilverNanoparticlesintoNeopreneFoamtoInhibit
BacteriaGrowth.

Kim,J.,Kuk,E.,Yu,K.,Kim,J.,Park,S.,Lee,H.,...Cho,M.(2006).AntimicrobialEffectsOf
SilverNanoparticles.
Nanomedicine:Nanotechnology,BiologyandMedicine,

3
(1),95101.

Lenntech.(2002).SilverAg.In
LenntechWaterTreatmentSolutions
.RetrievedfromINCHEM.
Shahverdi,A.,Fakhimi,A.,Shahverdi,H.,&Minaian,S.(2006).Synthesisandeffectofsilver
nanoparticlesontheantibacterialactivityofdifferentantibioticsagainstStaphylococcusaureus
andEscherichiacoli.
Nanomedicine:Nanotechnology,BiologyandMedicine,

3
(2),168171.
RetrievedDecember3,2014,from
http://www.nanomedjournal.com/article/S15499634(07)000469/abstract

Mijnendonckx,Kristel,NatalieLeys,JacquesMahillon,SimonSilver,andRobVanHoudt.
"Antimicrobialsilver:uses,toxicityandpotentialforresistance."
BioMetalsAnInternational
JournalontheRoleofMetalIonsinBiology,BiochemistryandMedicine
(2013).
Springer
StandardCollection
.Web.8Oct.2014.

Pradeep,T.,andPrashantJain."PotentialofSilverNanoparticleCoatedPolyurethaneFoamAsan
AntibacterialWaterFilter."
BiotechnologyandBioengineering
90.1(2005):5963.
WileyOnline
Library
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Sondi,Ivan,andBrankaSalopekSondi."Silvernanoparticlesasantimicrobialagent:acasestudyon
E.coliasamodelforGramnegativebacteria."
JournalofColloidandInterfaceScience
275.1
(2004):17782.
ScienceDirect
.Web.18Sept.2014.

WhatisNanotechnology?.(n.d.).In
NationalNanotechnologyInitiative
.

Protocol:


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Hauser,J.(2006,January1).TechniquesforStudyingBacteriaandFungi.RetrievedDecember16,
2014,from
http://www.carolina.com/pdf/caresheets/TechniquesforStudyingBacteriaandFungi.pdf

HowtoUseanInoculationLooptoTransferBacteria.(2014,October1).RetrievedDecember16,2014,
from
http://www.scienceprofonline.com/microbiology/howtouseinoculationlooptotransferbacteria.ht
ml

ReactivationofaMicroKwikCulture.(2013,January1).RetrievedDecember16,2014,from
http://www.carolina.com/pdf/caresheets/ReactivationofaMicroKwikCulture.pdf

Sanders,E.(2012,May11).AsepticLaboratoryTechniques:PlatingMethods.RetrievedDecember
16,2014,from
http://www.jove.com/video/3064/asepticlaboratorytechniquesplatingmethods