UV and Apple Cider Vinegar Interaction Effects on Flora Oscar Shaver Grade 11 Central Catholic High School
UVandAppleCiderVinegarInteractionEffectsonFlora
OscarShaverGrade11
CentralCatholicHighSchool
ElectromagneticSpectrum• Rangeofalltypesoffrequenciesofelectromagneticradiation• Thetypesofradiationareradiowaves,microwaves,infrared,visible,ultraviolet,x-rays,andgammarays
UVLightRays• Ultraviolet(UV)raysarelightraysthathaveshorterwavelengthsthanvisiblelight• Rangefrom150nm–300nm• Theyarenaturallygivenoffbythesun,butmostareabsorbedbytheozonelayer• Thewavelengthusedinthisexperimentis254nmlight
AppleCiderVinegar• Bragg‘sAppleCiderVinegar(unfiltered)Ø FermentedrawapplejuiceØ UsesfromcookerytohealthbenefitsØ Internetphenomenon- manyclaimsØ Contains:vitaminB,vitaminC,folicacid,magnesium,potassium,iron,calcium,andbiotin
ØpH:3.075
VitaminCandUVlight• FreeradicalsinducedbyUVexposureØ createsoxidativestressØ chainreactionofdamagingcells‘sDNA• VitaminCisanantioxidantØ inhibitsoxidationØ reducesoxidativestress
Staphylococcusepidermidis
• Grampositivebacteria• Partofnormalhumanflora• Foundonskin• Coccalbacteria• Whenincubated,formswhitecolonies
Purpose• TodeterminetheinteractioneffectsofapplecidervinegarandUVlightonStaphsurvivorship
Hypotheses
Null:ApplecidervinegarwillnothelptoincreasesurvivorshipofUVstressedStaph
Alternate:ApplecidervinegarwillhelptoincreasesurvivorshipofUVstressedStaph
Materials(DirectExposure)• YEPDagarplates(YEPDmedia+1.5%agar)
• YEPDmedia(1%yeastextract,2%peptone,2%glucose)
• Incubator• Ethanol
• Spreaderbars• 0.2MicronSterileSyringeFilter
• Staphylococcusepidermidis
• Bragg‘sAppleCiderVinegar(unfiltered)• UVlamp(254nmlight)
• Testtubes• SterileDilutionFluid[SDF](100mMKH2PO4,100mMK2HPO4,10mMMgSO4,1mMNaCl)
• Sterilepipettetips
• Micropipettes• Sterilecappedtesttubeswithsteriledistilledwater
• Sidearmflask
Procedures(DirectExposure)1. Bacteria(Staph)wasgrownovernightinsterileYEPDMedia2. Asampleoftheovernightculturewasaddedtofreshmediaina
sterilesidearmflask3. Theculturewasplacedinanincubator(37°C)untiladensityof50
Klett spectrophotometerunitswasreached.Thisrepresentsacelldensityofapproximately10⁸cells/mL
4. Theculturewasdilutedinsteriledilutionfluidtoaconcentrationofapproximately10⁵cells/mL
5. Applecidervinegarwassterilefilteredusing0.2micronsterilesyringefilter
6. ApplecidervinegarwasaddedtotesttubestocreatethefollowingpH’s:4and5
ConcentrationChartFinalConcentrationinTubes
0%(pH6.5) 2%(pH5) 5%(pH4)
SterileWater 9.9ml 9.7ml 9.4ml
Bacteria 0.1ml 0.1ml 0.1ml
AppleCiderVinegar 0ml 200microliters 500microliters
TotalVolume 10ml 10ml 10ml
Procedurescontinued7. 0.1mlofcellculturewasthenaddedtothetesttubes,yieldingafinal
volumeof10mL.8. Thesolutionsweremixedbyvortexing andallowedtositatroom
temperaturefor10minutes9. Aftervortexing toevenlysuspendcells,0.1mlwasremovedfromthe
tubesandspreadonYEPDplates10. PlateswerethenplacedunderaUVlampforcertaintimes(0,4,and8
seconds)11. Theplateswereincubatedat37°Cfor24hours12. Theresultingcolonieswerecounted.Eachcolonyisassumedtohave
arisenfromonecell
pHandRadiationExposure
00
L0
H0
0L
LL
HL
0H
LH
HH
6.5pH(0) 5pH(L) 4pH(H)
0sec
4sec(L)
8sec(H)
P-valueInteraction:0.035003
P-value:0.040385
P-value:1.47E-05
Procedures(LiquidExposure)1. Bacteria(Staph)wasgrownovernightinsterileYEPDMedia2. Asampleoftheovernightculturewasaddedtofreshmediaina
sterilesidearmflask3. Theculturewasplacedinanincubator(37°C)untiladensityof50
Klett spectrophotometerunitswasreached.Thisrepresentsacelldensityofapproximately10⁸cells/mL
4. Theculturewasdilutedinsteriledilutionfluidtoaconcentrationofapproximately10⁵cells/mL
5. Applecidervinegarwassterilefilteredusing0.2micronsterilesyringefilter
6. ApplecidervinegarwasaddedtotesttubestocreatethefollowingpH’s:4and5
Procedurescontinued7. 0.1mlofcellculturewasthenaddedtothetesttubes,yieldinga
finalvolumeof10mL8. Thesolutionsweremixedbyvortexing andallowedtositatroom
temperaturefor10minutes9. Aftervortexing toevenlysuspendcells,800microliters were
removedandaddedtomicrotubes10. Openedmicrotubes werethenplacedunderaUVlampforcertain
times(0,4,and8seconds)11. Afterexposure,0.1mlwasremovedfromthemicrotubes and
spreadonYEPDplates12. Theplateswereincubatedat37°Cfor24hours13. Theresultingcolonieswerecounted.Eachcolonyisassumedto
havearisenfromonecell
P-valueInteraction:5.27e-12
P-value:1.66E-05
P-value:7.5E-09
Conclusions
• Thenullhypothesiscanberejectedforeverygroupinbothdirectandliquidexposureexperiments• ItcanbeinferredthatapplecidervinegarhadaprotectioneffectagainsttheUVstressedStaph
ØApplecidervinegarandUVlightappearedtoreducestaphsurvivorshipindividually
Limitations
• Spreadplatingwasnotperformedinperfectsynchronization,thereforesomebacteriahadaslightlyhigherexposuretime• Onlyonespeciesused• Onlysurvivorshipassessed• Platingwasdoneinroomwithpartialsunlight
Extensions
• Testandcompareotherliquidscontainingvitaminc• Morespeciesofbacteriawillbeused• Spreadplatingwillbeperformedmoresynchronously• UVexposuretimewillbeatsmallerincrements
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Nov.2017."TypesOfUVLight- UVC,UVB,UVA ." AmericanAir&WaterUVLightAirCleanersand Ultraviolet
WaterPurifiers.N.p.,n.d.Web.24Jan.2018.“UV&FreeRadicals." Albus&Flora.N.p.,n.d.Web.24Jan.2018."VitaminCandSkinHealth." LinusPaulingInstitute.N.p.,01Jan.2018.Web.24Jan. 2018."WhatIsGermicidalUltraviolet?|Ultraviolet.com."Ultraviolet.com.N.p.,n.d.Web.23Nov.2017.
2FactorANOVA(DirectExposure)
2FactorANOVA(LiquidExposure)