TheFundamentalTheoremofAlgaebra

SeanJordan
BethDungey
ErinRadebe&RogelioZimbron
9March2016
IndependentResearchIG/T

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Abstract
Intodayssociety,thebenefitsofalgaearenumerous,butareoftenignoredinfavorof
easier,moreprofitablemethods.Sincethemajorityoftheworldsenergycomesfromfossilfuels
(suchascoal),thiscouldspelldisasterwhentheyinevitablyrunout.Themostrecognizable
benefitofalgaeisitsuseinbiofuel.Unlikefossilfuels,biofuelsarearenewablesourceoffuel
andenergy,whichmeansthattheycanbeusedoverandoveragain.Asofrightnow,biofuels
onlymakeupabout7%oftheworldsenergysupply(Funk,2015).Ifthispercentagecanbe
increased,therelianceonfossilfuelswillbedecreased,andwillinturnhelpwithfuture
fuel/energyproduction.Inordertohelpsolvethisproblem,astudywillbeconductedthat
determineswhichalgaestrainyieldsthemostamountofbiofuel.Also,researchwillbedoneon
whatmethodofharvestingalgaeforbiofuelismostefficient.Abenefitofalgaeoverother
sourcesofbiofuelistheirhighoilyield(Funk,2015).Thisstudyrequiresthatmultiplestrainsof
algaebeculturedandcentrifugedtofindouttheamountofbiofuelproduced.Thedetailsof
culturingandcentrifugingthestrainswererecordedinanotebookasatimelineofevents.The
resultsofthisprojectwillbesharedwithagroupofscientistswhoareinterestedinalternative
sourcesofenergy,and/orareinterestedinthepossiblebenefitsofalgaeintodayssociety.This
informationwillbesharedthroughadetailedwebsiteontheinternet.Thefindingsfromthis
projectwillteacheveryonetorecognizethebenefitsofalgaeandpossiblyencourageenergy
producerstoswitchtoarenewablesourceofenergy.

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Introduction
Theyarethemean,sometimesgreen,oxygenmachine.AccordingtotheOceanography
DepartmentfromtheUniversityofSanDiego,theyproducemorethanhalfoftheworlds
oxygensupply.Infact,thesemysteriousorganismshavebeenontheEarthmuch,muchlonger
thanthehumanpopulation.Theyareavitallinkinthefundamentalfoodchainduetotheir
photosyntheticproperties.Infact,theyareslowlybecomingamoreprominentfactorintodays
society.Thesemysteryorganismsarealgae.Algaecanbenefittodayssocietybybeingasource
forbiofuels,holdingthefoodchaintogether,andpossiblyhelpingdiseasedcells.
LiteratureReview
Themostlikelybenefitofalgaerecognizedbypeopletodayistheiruseinproducing
biofuels.Biofuelsareessentiallyoils,suchasethanolandbiodiesel,thatareextractedfrom
organicmatterandthenburnedtoproduceenergy(Top10Sources,2015).Accordingto
DiscoveryNews(2015),algaloilsarethesecondmostcommontypeofbiofuel,behindcellulose
fiber.TheBiofuelsUKwebsitestatesthattherearemanydifferenttypesofbiofuelsthatcanbe
synthesizedfromtheoilofalgaecells.Biofuelscanbeseparatedintofirstgenerationfuelsand
secondgenerationfuels.Firstgenerationproductsincludethesourcesthatarenotreadily
renewableortakealongtimetomake.Anexampleofthiswouldbetheworldsfoodsupply,
suchascornstalks,wheat,andanyothercrops.Thefuelsthatcanbemadefromfirstgeneration
productsincludebiodiesel,biogas,andcertainfoodoils(TypesofBiofuel,2010).Thebiggest
advantagetothismethodisthatitcouldpossiblyraisesocialandeconomicawareness.The
problem,though,isthatitcouldalsostartcuttingintotheworldsfoodsupply.Thiswouldentail
amajorfoodvs.fuelargumentthatwouldbecatastrophictooursupplyofbothfoodand

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energy(Naik,Goud,Rout,&Dalai,2010,pg.580).Withoverpopulationbecomingabiggerand
biggerproblem,thiscouldbecomeacontroversialissue.Analternativetothiswouldbesecond
generationbiofuels.Thesefuelsarenevermadefromtheworldsfoodsupply,andareinstead
synthesizedfromrenewableorganisms,suchasalgae.Theseorganismsreproducequicklyand
helpensurethattheenergydemandismet(Top10Sources,2015,para.1).Thesecond
generationsourceshavetheirownproblemsaswell,someofwhicharereflectedinalgae.For
one,whiletheydoreproducequickly,theymayreproduce
so
quicklyastoblockthelightthat
othercellsintheculturedesperatelyneedtosurvive.Thesourcefurtherstatesthatthisproblem
couldpossiblyberegulatedbyconsistentlytakingaportionofthealgaeoutofitsculturing
environmentforconversionintobiofuel.Thismethodhelpstoensurethatenoughlightreaches
eachcell,andnocatastrophiccelldeathoccurs.Asourceofalgaethatisworthmentioningis
eutrophiclakesandareas.Eutrophicareasareregionsthatarehighinnutrients,suchasnitrogen
andphosphorous.Thiscancausemassivealgalblooms,whichcanthenbecontinuously
harvestedforfuel(EutrophicationExperiments).Anotherbenefitthatalgaecellshaveover
bothfirstgenerationandsecondgenerationsourcesincludetheircomposition.DiscoveryNews
saidthatbecausealgaeareroughly50%fat,theyproduceanabnormallylargeamountofbiofuel
comparedtotheirsecondgenerationcousins(Top10Sources,2015,para.1).Thebiggest
problemtobeovercomewithusingalgaeforbiofuel,however,isthefactthatitusesavast
amountofwater.ThereismorethanonesextillionlitersofwaterintheEarthsoceans.The
processofconvertingalgaeintobiofuelusesapproximately3000litersofitperharvest
(Buchele,n.d.,para.5).Whatisevenmoreofaproblemisthatthisamountofwateronly
produces
one
literofusablebiofuel.Usingthisamountofwaterforsuchasmallamountoffuel

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isimpractical.However,ifthewatercanberecycledthroughtheharvestcyclesofalgae,thenthe
amountofwaterperharvestcanbedrasticallyreduced.Themainproblemwiththisrecycling
methodisthatitrequiresconstantmaintenancetomakesureenoughwaterstaysinsidethearea
andthealgaedonotkilleachotheroff(asaforementioned).Themaintenanceworkerswould
alsobeinchargeofharvestingthealgaeforbiofuel.Thisconstantmaintenancecannotonlyhelp
establishamajorfuelsource,butcanhelpemployhundredsofworkersattheproductionareas
whileconservingpotablewater(Siegel,2012).Allinall,algae,withthecorrectregulationsand
maintenance,areanotablesourceforbiofuelsintodayssociety.
Anotherhugebenefitofalgaeontodayssocietyisitsparticularlystrongroleinthefood
web.Algaeareatthebaseofthemarinefoodweb(FittingAlgaeintotheFoodWeb,n.d.,
para.4).Thismeansthattheymakeupthefoundationofthethousandsoffoodchainsthatare
formedwithinthethreedomains.Theyarewhatareknownasphotoautotrophs,whichmeans
theytakeintheenergyfromsunlightandconvertitintoconsumableenergy.Infact,algae
precededeventhecommonplantsweknowtodayinchlorophyllproductionandphotosynthesis.
Thisenergythengetspassedupthroughthefoodchain(witha~90%energyloss)tothe
herbivores,andeventuallythecarnivores(Ophardt,2003).Besidesthefoodweb,algaeareatthe
basisoftheevolutionofplants.Evidenceshowsthatamultitudeofsinglecelledorganisms,such
asredandgreenalgae,gaverisetothenonvascularplants,whicharetheonesthatmovedto
land(Segovia,Haramaty,Berges,&Falkowski,2003).Allofthissupportstheconclusionthat
algaeareavitallinkinthefoodweb,andthattheworldssocietyneedstheseorganismsto
surviveifthehumanraceistocontinueliving.

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Anotherpossibilityofalgaesbenefitsinthefoodwebisfoodforhumans.J.Craig
Venterbelievesthatwithalittlegenomicengineering,wecanspeedupevolutionbymillionsof
years(Biello,2011,para.6),andhopefullymakemeatfromtesttubesformedfromsynthetic
biology.Algaecellshaveahugepartofthis,asanastronomicalportionfromwhereheplansto
takethemicrobesarealgaeculturesintheocean.Anothergoodsourceoffoodisseaweed,which
floatsrightalongsidealgaeinthefoodweb(FoodWebCards,n.d.).Withthistechnology,the
worldsgrowingfooddeficitcanhopefullybesolved.
Specificstrainsofalgaehavedifferentqualitiesthatcanpossiblylendahandinother
areasofresearch.
DunaliellaTertiolecta
isonesuchstrain.Accordingtoastudyfromthe
QueensUniversityofBelfast,the
D.Tertiolecta
strainundergoesauniqueformofcelldeath
thatisstrikinglysimilartoapoptosisineukaryoticorganisms(Segovia,Haramaty,Berges,&
Falkowski,2003).Theseresearchersaretryingtofindoutmoreaboutthepossibleoriginof
apoptosis,andtheyreusingalgaetodoit.Apoptosis,alsoknownasProgrammedCellDeath,is
describedasanactive,programmedprocessofautonomouscellulardismantlingthatavoids
elicitinginflammation(Fink&Cookson,2005,para.4).Thisprocessiscommonlyusedduring
thedevelopmentofthefetus.Duringprenatalcellreproduction,geneshelpassigncertaincellsto
certainbodilyfunctions.Theseincludethewebbingbetweenfingersandtoesandatail(Alberts,
Johnson,Lewis,Raff,Roberts,&Walter,2002).Obviously,humansdonothavethesestructures
aspostnatalorganisms.Thesestructuresundergoapoptosisandareharmlesslyremovedfromthe
bodybywhitebloodcells.Accordingtothestudy,apoptosisineukaryoticorganisms,suchas
humans,evolvedfromprokaryoticorganisms,suchasthe
D.Tertiolecta
strain(Segovia,
Haramaty,Berges,&Falkowski,2003).Thereareothertypesofstructuresthatwedonotneed

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anymoretofunction.Thesestructuresarecalledvestigialstructures.Inhumans,theseincludethe
appendixandtailbone.Thesestructuresspecificallydonotundergoapoptosis(Vestigial
Structures,2016).Ofcourse,thisisnottheonlytypeofcelldeath.Theharmfultypeofcell
deathiscallednecrosis.Necrosisoccurswhenthereisanintrusionfromanunwantedobject,
eitherfromoutsidethebody(injuries)orinsidethebody(viruses).Whenthesecellsdie,they
burstopen,causinginflammatoryresponsesfromneighboringcells(
1.2Differencesbetween
necrosisandapoptosis
,n.d.).Beyondthis,algaecellshavebeenextensivelystudiedasto
provideinformationonendosymbiotictheory.AstudybyLynnMargulisattheUniversityof
Berkeleyshowshowalgaecellswereusedinordertohelpprovethetheory.Theystudiedthe
mitochondriainbothalgaecellsandeukaryoticcells,andnoticedmanysimilaritiesbetweenthe
two(Margulis,n.d.).Thus,itsupportsthenotionthatalgaeplayedahandintheevolutionof
prokaryoticcellsbypassingalongtheirmitochondria.Thisevidencecouldpossiblybeappliedto
diseasedcells(suchascancerouscells)inordertoremovethethreatfromthebody.
Thesecountlessoxygenmachinesareconstantlyhelpingoursocietyinmorewaysthan
one.Theycanhelpreducethenumberofnonrenewablefossilfuelsusedforelectricityandcan
aidinmeetingtheevergrowingelectricitydemand.Beyondthat,theyplayafundamentalrolein
thefoodchaininthat,theyareamajorfactorinthebaseofthemarinefoodweb.Thisenergy
fromphotoautotrophs,suchasmicroandmacroalgae,eventuallymakesituptothetopwherethe
humanraceisthoroughlysustainedduetotheseorganisms.Finally,researchhaspointedtoa
hypothesisofpossiblyusingtheapoptosislikecelldeathprocessinthe
D.Tertiolecta
strainto
helpcurediseasedandpossiblycancerouscellsineukaryoticorganisms,suchashumans.The
overallbenefitsofalgaearenumerous,andcanhelpoursocietyinunforeseenways.Ifthetop

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scientistscantakeadvantagesofthesebenefits,thentheworldwillundoubtedlybeabetter
place.

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DataCollection/Methods
Duringthisexperiment,theculturingenvironmentwassetupinMs.Herbersroom.It
wasintherearfumehood,whichmadeiteasiertomaintainastableenvironmentthatwasbest
suitedfor
DunaliellaSalina
.Therewerealotofresourcesnecessarytoexecutethisproject.
Therewasaneedforanairsplitter,anairpump,someairtubes,atimer,andacoupleofplastic
juicebottles.Inordertohelpwithdataobservation,amicroscopeandlabquestmachine(pH)
wereborrowedfromotherscienceteachersatLongReach.The
D.Salina
strainswerebought
fromCarolinaBiologicalSupply.
Whiletheexperimentwasgoingon,observations,data,andillustrationswererecordedin
alaboratorynotebook(AppendixA).Thisdatacollectionconsistedofupdatesontheproject,
dayswhentheculturewasbrieflyobserved(
checkin
),anddayswhentherewasanextended
periodoftimeperformingexperimentsonthealgae.
DataAnalysis
Throughoutthefirstcoupleofweeks,theobservationsindicatedthatthealgaeculturing
wascomingalongnicely.Thesurroundingmediumsteadilyturnedadeepershadeofgreen,and
theclumpsfromthestartingvialslowlydissolved.TheculturealsosurvivedoverThanksgiving
Break,whichwasamajoraccomplishmentwithouthumansupervision.Becauseofthis,itwas
thoughtthatthealgaewoulddofineoverwinterbreakaswell,providedthattheschool
custodianswouldmaintainthecurrenttemperature.Oncebreakwasover,basicobservations
indicatedthattheculturehadsurvivedovertheweek.Afterthis,though,therewasnonotable
changeinthealgae.Then,agiganticsnowstorm,alsoknownasBlizzard2016,hit.Asbefore,

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theresearcherthoughtthattheschoolwouldmaintainitstemperature,likeitdidoverwinter
break.Whentheweekofnoschoolpassed,theresearcherstartednoticingweirdbrownspotson
thesideoftheculturingenvironment.Thesewereinfactdeadalgaecolonies,andtheywerea
telltalesignthatsomethingwasnotright.Then,theresearcherwasinformedbyhisadvisorthat
theschooldidntactuallymaintainaconstanttemperatureoverthesnowweek.Atthistime,the
twoadvisorssuggestedtryingtocultureitonemoretime,astheendoftheyearwasrapidly
approaching.Afellowalgaecultureragreedtohelp,andhehadamoreadvancedculturing
environment.Therewasalmostacertaintythatthistimewouldbeasuccess.Thistime,however,
thealgaediedalmostimmediately.Becauseofthis,theresearchersandadvisorscametothe
conclusionthattheproblemwasthealgae,nottheenvironment.Afterfurtherresearch,itwas
discoveredthatthe
D.Salina
strainisoneofthehardeststrainstoculture,duetoitshighpH
requirement.Afterthisincident,itwasdecidedthattheprojectwouldmakeuseofthefellow
algaeculturersstockcultures.

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AppendixA
November11:
Ifinallyfinishedsettinguptheculturingenvironmenttoday.
Ihadtoborrowtheclamplight(thethingthatholdsthephotosyntheticlight)fromMr.
Emerick.
Ialsosterilizedtheplasticbottleswithpureisopropyl(typeofalcohol).
FollowingMrs.Radebesinstructions,IsetthetimerIboughtto12hrs.on/12hrs.off.
ThelastthingIdidwastosetuptheairnetwork.Thisinvolvedconnectingairtubesto
theairpump,theairsplitter,andthebottles.
November18:
Thealgaeandmediumweredeliveredtothemediacentersometimetoday,andIpicked
themup5thperiodtoavoidanydeathinthecells.
Idepositedthemediumandcultureintothefirstbottle,andasofrightnowIcanonlysee
bitsandpieces.
November24(Checkin):
Thealgaemixturelooksslightlygreener,whichIguessisagoodsign.
ImadesomeadjustmentstoincreasethechanceofsurvivaloverThanksgivingBreak.
Thisincludesadjustingtheangleofthephotosyntheticlightandchangingthetimerto15
hrs.on/9hrs.off.
November30(Checkin):

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Atfirstglance,itappearsthatthealgaestrainsurvived.Itsstillgreen,andIcanseemore
particlesinthebottlethanbefore.
OnceIfoundasecondadvisor(RogelioZimbron),hesuggestedIlookatthealgaeunder
amicroscopeassoonaspossible.
December7:
ThoseparticlesthatItalkedaboutaremoredense,whichmayimplythatthereare
concentratedculturesinthebottle.
AsperMr.Zimbronsrequest,Iamlookingatacellunderamicroscope(whichI
borrowedfromMr.Renehan).
Iusedasterilepipettetopickupasmallportionofthestrainandplaceitonawetmount
microscopeslide.
Ifoundanalgaecellunderthemicroscope.Itappearsasifitsdead(oritsanimmobile
strain).
Now,accordingtoMrs.Radebe,allIneedtodoischeckinonthealgaeforamonthor
so.
Ihaveafeelingthatthisprojectmayextendintomyjunioryear,ifIdontdropitoverthe
summer.
ThisincludesthechallengeoftakingIndependentResearchorIntern/Mentornextyearor
not.

December11(Checkin):

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Mrs.RadebeandIhavedecidedtostartanewsubcultureafterWinterBreak,providing
itsurvivesaweekwithoutanyonecheckinginonit.
December14:
Icanseeagoodnumberofconcentratedculturesstickingtothebottomofthebottle.
Itriedshakingitup,butnothingchanged.Iprobablyneedtodoitmorevigorously.Ill
talktoRadebeaboutit.
IllalsoseeifIcanlookatanothercellunderthemicroscopesoonerorlater.
December21(Checkin):
Thereseemstobenochangeinthealgae..Ihopethatthestrainisstillalive,ifiteven
wasinthefirstplace.
Ifitisalive,thenthereistheproblemofWinterBreak.Idontreallyknowwhattodo..I
couldeitherleaveithereandhopethetemperatureinsidestaysthesame,ortakeithome
andriskinfection.
IwillaskRadebeaboutit,butsheisnowheretobefoundrightnow.
Ishookthebottleupmoreroughlythistime.Therewasaslightchange,butnothing
major.Thismeansthatthecellsarestickingtothebottom,whichIdontthinkisagood
sign.
Iaddedtheremainingportionofthemediumtothebottle,asAaronhasntusedityetfor
hisculture.
December22(Noncheckin):
Ihavedecidedtoaskthebuildingsupervisor(Mr.Baylor)aboutthebuilding
temperatures.

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January4:
Nochanges.Ithinktheculturemaybedead.
Ipossiblysawanothercellunderthemicroscope.
January8:
AsperMr.Zimbronsrequest,IamtestingthepH
ofthemediumwiththelabquestmachine.
Thefirstreadingwaswiththeairpumpoff.Itread
5.5,whichisacidic.
Thesecondreading,withtheairpumpon,sawa
driftinthepH,constantlygoingeversoslightlytowardsalkalinity.
ItfinallystoppedatapHofaround7.80
January11(Checkin):
Strangereddishbrownishspotshaveappearedonthesidesoftheplasticbottle..Idont
knowwhattheyare,andIlltryandlookatthemunderthemicroscopewhenIgetthe
chance.
January15(Checkin):
Stillnochangeinthealgae.
Ithinksomethingswrong.
AaronPark,afellowalgaeculturer,hasinformedmethatthereddishbrownishspots
wereinfactdeadcoloniesofthestrain.
Hecheckedthetemperatureofthemedium,anditseemedfinetohim.

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Hedecidedtomovemyculturetohisenvironmentinaglassbottleinadesperateattempt
tosaveit.
February10:
Sothealgaedied.AsIlearnedonTuesday,theschooldidntkeepthetemperatureupto
parduringBlizzard2016.Werecurrentlydebatingwhattodo.
Wedecidedtotryandorderthealgaeagain,andhopefullymakemyownmediumthis
time.ThealgaerequiresapHofapproximately9,whichisquiteodd.
Ishouldhaveabout9weeksoftestingafterthealgaearrives(ifitsurvives).
February19(Noncheckin):
MeandAarondecidedtostayafterschoolonedaythisweektocustommakeamedium
formyalgaestrainthathasahighsaltconcentration.
February26(Noncheckin):
MeandAaronstayedafterschooltocustommakethemedium.
ItstheModifiedJohnsonsMedium,whichhasahighsaltconcentration.
Thealgaestillhasntbeenordered,astheteacherthatcanorderitiscurrentlyabsent.
Wereprobablygoingtoorderthealgaetogetherandsplitthecostsofthereceipt
accordingly.
March9:
Thealgaeissettoarrivetoday,andMrs.Radebeisnthere.
March12:
Thealgaediedimmediatelythistime.AaronsaidthathewillcallCarolinaandaskfora
refund,astheydidntmentiontherequiredpHofthe
DunaliellaSalina
strain.

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March24:
IvedecidedtojustuseAaronsalgae,ashisculturedsuperbly.Ihopehisstrainswill
surviveoverbreak.
April4:
Aaronhasinformedmethatthestrainssurvivedoverbreak,andthatImgoodtogo.Ill
tryandpushthroughitquicklysoIcanputinformationonmydisplayboard.

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