JoshuaLubben
TheEffectivenessofRiceasaCost-EffectiveSoilBacteriaCaptureMethod

Soilisanimportanthabitatfororganismswitchprovidemanyimportantfunctionstonaturalsystems.Multiplestudiesshowingthatsoilfungihelpplantsaccessandtakeupvitalnutrients(Halifuetal,2019;Chen,etal.2020;Kang,etal.2021).Thesenaturalfunctionshavebeenfoundtotransferovertoagriculturalsystemsaswell.Wissuwaandotherresearchers(2020)foundthatamicrobialfunguswasfoundtohelpriceplantsaccessphosphorusindeprivedsoils.Pepperplantsthatwereconnectedtoalargenetworkoffunguswerefoundtohaveaheighteneddroughttolerance(Marasco,etal.2012).However,formanydecadesconventionalagriculturalpracticeshavebeenunsustainable.Thesepracticesrelyheavilyonartificialmanipulationoftheenvironmenttokeepthesoilproductive.Thecommonpracticeoftillingdisturbssoilstructureandcausesmicrobialcommunitiestochangeinpopulationstructure(Smith,etal.2016).Growingthesamecropyearafteryeardepletesthesoilofnutrients,requiringtheadditionoffertilizers.Growingthesamecropsalsocausesalargerpopulationofspeciesthatfeedoffthecrop,sofarmersneedtousepesticidestocontrolpestpopulations.Inthepastfewyears,theideaoforganicagriculturehasgrowninpopularity.Thismethodofagriculturelookstorelylessontheconventionalwaysofagriculture,andmoreonnaturalproses.Asorganicagriculturerelieslessontillageandchemicalsitlookstonaturesystemsforinspiration.Thatmaybeplant-basedfertilizers,companionplantingormanyothermethods.Oneimportantpracticeoforganicagricultureistheuseorbeneficialorganismscommonlyreferredtoasbeneficial.Beneficialprovidemanyfunctionssuchaspopulationbybeesandbutterfliesandpredationofpestsbygroundbeetlesandladybugs.Onesubsetofbeneficialsiseasilyoverlooked,microscopicsoilorganisms.Duetodegradedandalteredsoilssoillifecanbeinapoorcondition.Exceptionallyifthesoilhasbeenunderconventionalagriculturalpractices.Tillagedisruptssoilstructureandcausesachangeinpopulationofsoilmicrobes(Smith,etal.2016).Soitmaybenecessaryforsoilmicrobestobeintroducedtothesoiltoetherchangediversityorreintroducevitalspecies.Itispossibletobuysoilmicrobialstartersbutthereisnoguaranteethatthosemicrobesarenaturalizedtoasystem’snaturalprosses.Thecostofbuyingthesestartersmaybeinefficientforsmallproducersandindividualsgardeners.Onepossibilityisusingmicrobesfromalocallocation.Therenativeandnaturalizedmicrobeswillbeusedtonaturalprossesintheenvironment.Reddy(2011)describedaprocessestocapturelocalmicrobesfromanundisturbedhabitat.Thecapturedmicrobescanthenbetransportedandspreaduponorganicagriculturalsystems.Reddy’sprocessesinvolvesleavingaboxofboiledwhitericeinalocalfieldorforestwherethelocalmicrobesmoveintofeedupontherice(2011).Thisstudywillusethismethodofsoilmicrobecaptureanddeterminehowthismethoddoesincapturingthetruediversitywithinthesoilbiome.Thesecondobjectiveofthisstudyisifthereisanydifferencebetweenusingwhitericeandanativespeciesofrice.ToevaluatethesequestionsfourplotswillbesetupinthefieldsandopenareasofLakeSuperiorStateUniversity’sHuntCreekFieldStation.Eachplotwillconsistofthreeboxessetuponbaresoil.Theboxeshavethinbottomswithholesdrilledinthemtogivesoilmicrobesaccesstotheinside.Eachplotwillreceiveoneboxofboiledwhiterice,oneboxofboiledwildriceandoneboxofcleanpottingsoil,whichwillactasacontrol.Asoilsamplewillbetakenfromeachplot.Theboxeswillbeleftinthefieldforfivedays.Attheendofthetime,eachboxwillhavesamplestakenoutofthem.Thesampleswillberunusingthe16sMetagenomicstest.Thistestwasusedbymultipleotherstudiestogathersimilardata(Valverdeetall,2016;Sumertaetall,2020).Thiswillgivethepercentagesandnumberofspeciespresentineachsample.Thetestfromtheboxeswillbecomparedtothetestresultsfromthesoilsamplestaken.Themaininformationthatwillbelookedfor,ishowcloselyaboxrepresentsitscorrespondingsoilsampleandisthereanystatisticallysignificantdifferenceinthevaluesbetweentheboxesandthenaturalsoilamongtheplots.

ChaseJannetta
DesignandExperimentationofaHighAccuracyVision-BasedAdjustableProjectileLauncher

 Vision-basedfeedbackisanongoingfieldofresearchinroboticswithmanyapplications.Visionsystemsareusedwidelyinapplicationssuchasself-drivingcars,factorylines,andaerialanalysis.Self-drivingcarsthatusevisiontodetectpedestrians,trafficandmakedecisionsbasedonthevisionfeedbackarebeingdeveloped(ÉloiZablocki,2022).Quadrotorsthatusevisiontonavigatearebeingresearchedandtested(Foehn,2022).ThegoalofourprojectistodevelopthesystemneededtoaccomplishTask7oftheRobotXCompetitionusingvision-basedfeedback.Task7involvesidentifyingthecorrectdockwiththetarget,navigatingtothatdock,andlaunchinga2-inchdiameterracquetballthroughaholeinthetarget(Robonation,Task7-FindandFling,2022).Tothisend,aprojectilelauncherwillbedeveloped,capableoflaunchingthefourracquetballsprovided.ThelauncherwillbeinterfacingwithTeamAMORE’sWAM-V,anunmannedsurfacevesselequippedwithnavigationandvisionsystems(TeamAMORE,2022).CodewillbedevelopedthatallowstheWAM-VtoidentifytherequiredtargetviatheonboardZED2icamera,andnavigatetothattarget.Astheballislaunchedatthetarget,vision-baseddatawillbecollectedusingthecamera,aboutthepositionoftheballasiteitherpassesthroughthegoal,orasitmissesthetarget.Thisdatawillbeusedtoadjusttheonboardsimulationcontrollingthelauncher’sfiringvelocityandreleaseangle,inordertoincreaseitsaccuracy.Doingso,themoreshotsarefired,themoreaccuratetheshooterwillbe.

GabrielleMinier
TheEffectsHostHaveontheMicrobiomeof MichiganTicks

TheobjectiveofthisresearchprojectistoseeifthereisanyrelationshipbetweenthelasthostofMichiganticksandthebacteriaintheirmicrobiome.

CarrieGregg
PresenceofTaxoplasmosiswithinFelinesandCaninesintheUpperPeninsulaAnimalShelters

LookingtoseeifToxoplasmosisisprevalentinSaultSteMariewithinfelinesandcaninesattheanimalshelter.ToxoplasmosisisanillnessthatcomesfromtheToxoplasmagondiiparasite,alsoknownas“CrazyCatLady”syndrome.Thisparasitehastheabilitytochangethebehavioroftheintermediatehoststobeeatenbythedefinitivehost.ToxoplasmagondiiinfectstheFelidaefamily,becausetheyproduceabunchoflinoleicacidthatisnecessaryfortheparasitetofullymatureandcompleteitslifecycle.Theparasitecanenterthebrainofafetusifapregnantwomanisinfected,anditcancauseadverseeffectsforimmunecompromisedindividuals.Knowingtheprevalenceinourareawillhelptobettermanageriskstothisparasite.

KiraKohler
ComparingCognitiveEffectsofPharmaceuticalAnxietyMedicationswithHolisticAlternatives

Pharmaceuticalanxiolytics,whilebeingdecentlyeffective,alsocomealongwithaplethoraofneurologicalsideeffectssuchaslethargy,sedation,ataxia,andevenseizures(BallantyneandCiribassi,2014;Sargisson,2014).ThemostnotoriousamongthesemedicationsisTrazodone,aselectiveserotoninreuptakeinhibitor(SSRI)whichhasbeenknowntocommonlycausestressonthecardiovascularsystem(Atlietal.,2019).BecauseTrazodoneisanSSRI,patientsthatusethismedicationarealsoatriskofdevelopingserotoninsyndrome,whichcanbedeadlyinextremecases(BoyerandShannon,2005).Duetotheadverseeffectsthatpharmaceuticalanxiolyticscanhaveonanimals,manypetownershavebeenlookingtowardholisticalternativestoeasetheirpets’anxiety,suchasmelatoninsupplements.Addingmelatoninsupplementstoananimal’sdiethasbeenobservedtoreducesituationalanxietyandeveninhibitmemoryimpairment(Lamtaietal.,2021).Consideringthefactthatpharmaceuticalmedicationshaveneurologicalsideeffectsthatareobservableeventotheuntrainedeye,therecouldbeapossibilitythatthemedicationisdoingmoreharminmoresubtlewaysthatownersmaynotnotice.Thepurposeofthisstudyistodetermineifthesepharmaceuticalanxiolyticsdo,infact,hinderthecognitiveprocessesofanimalsbycomparingthemtoholisticShouldthesemedicationsprovetoalterthecognitiveabilityofthesubjecttakingthem,thiscouldaffecttheoveralltreatmentforanxietyindomesticpets.Othermethodsoftreatmentwouldneedtobesoughtouttoreplacepharmaceuticalanxiolyticsforallanimals.Inordertotestifapharmaceuticalanxiolytic,specificallyTrazodone,hasanyeffectonthecognitiveprocessesofananimal,18Long-Evansratswillbedividedintothreegroupsofsixsubjects.Onegroupwillactasacontrol,anothergroupwillbeadministeredTrazodone,andthelastgroupwillbeadministeredmelatoninsupplements.Beforeeachgroupistreatedwiththesemedications,baselinetestingwillbeperformed,whereineachrats’workingmemory,spatialmemory,andmentalhealthwillbeassessed.Afour-weektreatmentperiodwillfollow,andafterwardtheratswillbetestedusingthesamemethodsasthebaselinetesting.Oncethedatahasbeencompiled,itwillbedeterminedwhetherornotpharmaceuticalanxiolyticsnegativelyaffectthecognitiveprocessesofanimals.

NaomiAstorga
PlantTumorCells’EffectsonGlycolysis

Summary:Oneofthehallmarksofcancerisanincreaseinglycolysisinthepresenceofoxygen(Jiang,2017).ThisisknownastheWarburgEffect.Thoughitisnotfullyunderstood,itisthoughttoresultfrommitochondrialdysfunctionofthecancercell.Iwouldliketotakeacloserlookatthiseffectusinganovelmodelplantdisease,termedcrowngall.Inthisexperiment,IwillbeinoculatingsoybeanplantswithAgrobacteriumtumefacienstoinducetheformationofcrowngalltumorsandquantifyingadifferenceofthelevelofpyruvicacidbetweentheuninoculatedplants(controlgroup)andthegroupsofinoculatedplantswithvaryinglevelsofinoculation.Introduction:Bothplantsandanimalcellshavemitochondriathatworktoproduceenergyforthecell.Inthepresenceofoxygen,plantswillundergocellularrespirationtoacquireATP,whichisusedforamultitudeofcellularprocessesrequiringenergy.However,whenoxygenisnotavailablethecellwillundergoglycolysisinthecytoplasm.GlycolysisisamuchlessefficientwaytogainATPasitonlyproduces2ATPwhereascellularrespirationoccursinthemitochondria,thepowerhouseofthecell,andproduces30-32ATP.TheWarburgEffect,orWarburg’shypothesis,statesthat“cancercellschooseaerobicglycolysisas[a]mainmodeofglucosemetabolisminsteadofmoreefficientoxidativephosphorylation”(Jiang,2017).Canceristheuncontrolleddivisionofcellsthatformmassesoftissuecalledtumors.Cancercellsalsohavedysfunctionalmitochondriathatcausesthecelltorelyonglycolysisthatcanbedoneinthecytoplasmoutsideofthemitochondria(Fadakaetal,2017).Agrobacteriumtumefaciensisagram-negativebacteriafoundinsoilthatcausescrowngalltumorsatwoundsitesoftheinfectedplants(Hooykaasetal,1982).Thebacteriumenterstheplantthroughwoundsitesandtransformsthehealthyplantcellsintotumors(Leeetal,2009).Soybeanplantsareaplantthattakeapproximately45-55daystomature.Theyrequirewarmsoiltogrowandgrowbestindaytimetemperaturesaround70°F(21°C).Theyarepoorsoiltolerantandrequireminimalspaceusageforpottingandsoil(McConnell,2019).Pyruvate,orpyruvicacid,isproducedbythefinalstepofglycolysisandcanbeusedtomeasurehowmuchglycolysisisbeingundergoneinasample.LactateDehydrogenase(LDH)-basedSpectrophotometricassayisthemostcommonwaytotestpyruvateconcentrations.Itisawell-establishedassaycompatiblefortakingthesemeasurementsfrommysamples.Importance:Therecouldbeacorrelationbetweenbacteriainfectinganorganismthatcausestumors(cancerous)cellsandthelevelofdysfunctionofthemitochondria.Thisisimportantbecauseofalltheotherpossibleimplicationsandquestionsthatfollowafterward.Ifthereisanincreaseinglycolysisthatcorrelateswiththelevelofinoculationofthebacteria,furthertestingcouldbedonetounderstandthecorrelation.Isthereacorrelationbetweentheamountofbacteriainoculumandtheamountofglycolysis?Howdotheglycolysislevelscorrespond(ordon’t)withtheactualtumorformation?Theseareimportantquestionsforunderstandingwhenthecell’smetabolic“transition”occursrelativetotheactualcancer(tumor)formation.Methods:Therewillbefourgroupsofthreeplantseachsoallmeasurementswillbedoneasameanofthetriplicate.Agroupofthreeplantswillhavenobacteriainoculationandwillbeusedasacontrol.Othergroups,allofthreeplantseach,willbe:single-woundinoculation,double-woundinoculation,andtriple-woundinoculation.Thiswillbringthetotalofplantstestedto12whichfitswellona96-wellplateusedforspectrophotometrywhichwillbeusedtomeasuretheabsorbanceandcalculatethepyruvateconcentration.Thesoybeanplantswillbegrownintheuniversitygreenhousetoensurethemostcontrolledenvironmentpossible,toincreaserepeatabilityandreplicabilityoffutureexperiments.Plantswillbegrownfor55daystoensuretheyhavereachedmaturity(McConnell,2019).AsmallincisionwillbemadealongthestemoftheplantandthewoundswillbeinoculatedwiththesamerelativenumberofbacteriacellsofAgrobacteriumtumefaciens,usinganinoculatingloop.AgrobacteriumtumefaciensisaBiosafetyLevel1(BSL-1)organism,soitiscompatibleforsafeusebystudentresearchers.Additionalincisionsandinoculationwillbedonetocreatethevaryinggroups(onewound,twowound,andthreewounds).Avisiblegalltumorcanbeseenin4weeks,soplantswillbemonitoredfor4weekstoensurethepropernumberofcrowngalltumorshavegrownforeachinoculatedplant(Gonzalez-Mulaetal,2019).ThepyruvateextractionprocedurewillbethemodifiedproceduredonebyZhuetal(2010)thatwasadaptedfromtheoriginalproceduresfromO’Donell-Tormeyetal(1987)andLamprechtetal(1974).Astandardcurveofabsorbanceandpyruvateconcentrationwillbecreatedusingknownconcentrationsofpyruvateextract.Theabsorbancewillbetakenat339nm,LDHwillbeadded,andthesamplewillbeincubatedatroomtemperatureinthedarkfor30minutesbeforeabsorbanceistakenagain.Thechangeofabsorbancebetweenthesetwomeasuredabsorbanceswillbeusedtocalculatethepyruvateconcentrationforthestandardcurve.ExtractionofLDHwillbedonebytheblendingofthesoybeanplantstohomogenizethem.Cellswillbeculturedandtreatedwithextractionbuffer,centrifuged,andputthroughacolumntopurifytheLDH(Rivoaletal,1991).TheLDHassaywillberepeatedonthecellsafterpurificationtreatment.Absorbancewillbetakenandusedwiththestandardcurvetocalculatethepyruvateconcentration(Zhuetal,2010).

EmilyMcDonald
ConversationswithDogs: DoesTalkingtoourCanineCompanionsReduceTheirStress?

Humansandcanineshavelivedtogetherforthousandsofyears,butthebondsthatonceweresimplycompanionshiphavebecomerelianceononeanother.Theanatomicalstructureofadog’sfaceandtheexpressionstheyusehaveevenbeeninfluencedbytheirrelationshipswithhumansbyevolvingtomimicthoseoftheirhumancounterparts(SmithandValkenburgh2020).Thesehavebeenrecognized,sometimessubconsciously,byhumansinneedwhentheyinteractwiththerapydogs.Thesimplepresenceofcanineshasbeenshowntoreducehumanstressinsituationsvaryingfrompatientsinemergencyrooms(Kline,Fisher,etal.2019)tooverwhelmedcollegestudents(FioccoandHunse2017).However,humansarenottheonlyoneswhosestresscanbereducedfrominteractionswiththeotherspecies.AstudyconductedintheUnitedStatesfoundthatshelterdogswhowerefosteredovernighthaddecreasedstresslevels,basedonbehavioralcharacteristicsandcortisol:creatinineratiosintheurine,whencomparedtonightstheyspentwithouthumancontactintheshelter(Gunter,Feuerbacher,etal.2019).Anotherstudyfoundthatpettingdogsaftertheyunderwentablooddrawreducedthestresslevelsoftheanimalduringasubsequentblooddraw20minuteslater(Hennessey,Williams,etal.1998).Interactionssuchastheseshowthatthoughanemphasisisplacedontheemotionalsupporthumansreceivefromtheircaninecompanions,thedogsfeelthebenefitsoftherelationship,aswell.Manystudieshavebeenconductedtodiscovertheimpactofourinteractionswithdogs,butmostfocusonthedirectcontactbetweenthespecies,suchaspetting.Fewtonostudieshaveexaminedtheimpactofday-to-dayvocalizationswithcaninecompanions.Theobjectiveofthisstudyistoexaminehowhumanvoiceandconversationimpactsthestresslevelsofdogs.Thehypothesisisthathumanvocalizationswillreducethebehavioralsignsofstressindogsaswellaslowertheurinecortisol:creatinineratio.MethodsAtotaloftendogswillparticipateinthestudy,allofwhichwillbefromeasternUpperPeninsulaanimalshelters.Aveterinarianwillbeonsitewhiletheresearchisbeingconductedtoensurethedogsremainhealthyandhandledcorrectly.Sincestayinginakennelhasbeenfoundtocausestressindogs,thiswillserveasanaturalstressorfortheexperiment(Hennessey,Williams,etal.1998).Afterbeingkenneledfortwohours,eachdogwillspendtenminutesinteractingwithahuman.Abaselinebehavioralassessmentwillberecordedduringinitialinteractionwiththedogtocomparetothefinalresults.Salivation,panting,pacing,andvocalizingwillbegradedonascaleof0-5,withzerosignifyingthatthebehaviorisnotpresent,andfivesignifyingthatthebehaviorisexcessive.Allofthesebehaviorshavebeenrecognizedasvisiblesignsofstressincanines(Almond,2018).Duringthetenminutesonthefirstday,thedogswillbespokentoinanormalvoiceandpetted.Thesamephrasestogreetthedogsusingtheirspecificnames,discussingtheplanfortheday,etc.willbeused.Afterthetenminuteinteractionwithvocalization,thedogwillbewalkedoutsidetovoidtheirbladder,andaurinesamplewillbecollected(free-catchusingacollectioncup).Thesamplewillbelabeledwiththetimeofcollection,date,nameofthedog,and“vocalization,”andwillbefrozenuntildataanalysisoccurs.Thesamebehaviorsasbeforewillagainbeobservedandrecordedafterthisproceduretodetermineifsignsofstressdecreasedafterthehumaninteraction.Thisprocesswillberepeatedforeachparticipantonthreeseparatedays.Toobtainacontrol,thehumaninteractionwillincludenovocalization,andthesameprocedurewillbeusedtorecordstressbehaviorsandcollecttheurinesample.However,onthesedays,thedogwillnotbespokentoatall.Interactionwillbelimitedtohumanpresenceanddirectcontact(petting).Thiswillalsooccuronthreeseparatedays,resultingindatafromeachdogforsixdays,threewithvocalization,andthreewithout.Forurinalysis,thecortisol:creatinineratiowillbemeasured.Methodsforthisanalysiswereusedinastudyconductedtodetermineifthisratioisavalidmeasurementofincreasedstressindogs.Thestudyfoundthatbymeasuringtheamountofcortisolusinganimmunoassaykitandcomparingthatvaluetotheamountofcreatinineinthesample(determinedbyJaffereactionandspectrophotometry),theratiowasavalidindicatorofstressincanines.Cortisolisahormonethatisreleasedwhencaninesarestressed.Anincreaseinthecortisolpresentinurinecorrelatestoanincreaseoffeelingofstressinadog.Thecreatininecontentisusedasameasureofurineconcentration,whichvariesbyanimal,hydration,andotherfactors.Bycomparingcortisollevelstothemeasureofcreatinine,theeffectsofconcentrationcanbeeliminated.Spectrophotometry,themeasureofasolution’slightabsorption,willbeusedinthisexperimenttoanalyzetheurinesamplesforbothcortisolandcreatinine(Part,Kiddie,etal.2014).Thesevalues,aswellasthevaluesassignedbasedonstressbehaviorswillbecomparedforeachindividualdogforvocalizedinteractionsversusinteractionswithoutvocalization.Ifthecortisol:creatinineratioandstressbehaviorswithvocalizationarelowerthanthevalueswithoutvocalization,itcanbedeterminedthathumanspeechandconversationlowerscaninestress.

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Spring2022GrantAwardees

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JeremyTurnbow
TheSpreadofBacterialKidneyDiseaseThroughCommonBaitFish

Renibacteriumsalmoninarum(RS),thebacteriumthatcausesBacterialKidneydisease(BKD),hashadasevereimpactonSalmonidpopulationsintheGreatLakessinceasearlyasthe1950s.Inthe1970s,BKDinfectedalargepercentageoftheSalmonidfishandcausedanalarmingnumberofmortalities.Tocombatthis,hatcheriestestforBKDineggsduetoitsabilitytotransfervertically.Sincethen,BKDhasbecomelessprevalent,butitisstillanendemicthreattosalmonidsintheGreatLakes.AfterhearingaboutBKD,IwonderedifitwaspossibleforBKDtotransferhorizontallybetweenspeciesorpossiblybeenintroducedtotheGreatLakesbyotherlivebaitfishviafishingactivities.Inparticulartomyproject,Iwanttoseeifcommonbaitfish,suchastheGoldenShiner,aresusceptibletoBKDandiftheyareabletotransferthediseasebacktoSalmonids.Iwillcollectbaitfishfromalocalbaitshopandtheywillbeplacedinto3tanks.Tank1willcontain10fishandwillnotbeinoculatedwithRS.Tank2willcontain10fishthatareexposedtoRSthroughthewater.Tank3willcontain10fishthathavebeenindividuallyinoculatedwithRSviainjection.Thefishwillbeacclimatedandthenfeddaily.Tankswillhaveafiltertoensuretheyhaveampledissolvedoxygen.Undertheseconditions,thefishwillthrivefor4-5weeksbeforebeingeuthanizedandtestedforBKDinthekidneytissuewithELISAkits.Ifthesefishtestpositive,wewilltheninoculate5moreGoldenShinersandhavethemliveinthesametankwith5AtlanticSalmonjuvenilestoseeifbaitfishcantransferBKDhorizontally.Ifthishappens,thismeansthatbaitfishacrosstheGreatLakescanbedamagingtoSalmonidpopulationsifinfectedfisharesoldtoanglers.

SophiePaquin
AStudyofNestingPreferenceandEffectivenessofNaturalReedsvsReusableWoodTraysinManagedSpringMasonBees,Osmiaspp

Withtheever-growingawarenessofthedeclineinpollinatorspeciesintheUnitedStates,whichisprimarilyduetohabitatlossbutalsopartiallylinkedtopesticideuseanddiseasespreadbytheEuropeanHoneyBee,agrassrootsmovementtoprotectnativebeeshasarisenintheformofprovidingnestingsitesfornativepollinators.Theseman-madenestingsitesarecommonlyknownas“beehotels,”orartificialdomiciles,andtheyaredesignedtoencouragenestingandpollinationbynativepollinators.However,asthetrendhasbecomemorepopularandtheneedmoreurgent,debateshavearisenwithinthenativebeeraisingcommunityoverthebestwaytogoaboutrearingnativebees.Ontheforefrontofthedebateiswhichofthetwopopularcommercialnestingmaterials,naturalreedsorreusablewoodtrays,aremoreeffectiveinpropagatingnewbees.Whilemanyobservations–andopinions–canbefoundononlinebeeforums,fewscientificstudieshavebeenconductedaddressingthequestion,withnonefocusingspecificallyonspringmasonbees,Osmiaspp.,themostpopularnativesolitarybeerearedintheUnitedStatesforfruittreepollination.Thegapintheliteratureiswide,andfindingsfromthisstudycouldbeconsequentialtoconservationandagriculturealikeasthe“SavetheBee”campaignbroadenstoincludemorethanjusthoneybees.Myseniorresearchideaexaminestwohypotheses:Naturalreedsaremoreeffectivethanreusablewoodtraysinraisingspringmasonbeesif“effective”isdefinedas“producingthehighestnumberofviablemasonbeecocoons.”Giventheoption,springmasonbeeswillshowapreferencetowardnaturalreedsoverreusablewoodtrays;theywillproducesignificantlymorecocoonsinreedsthantrayswhenbothoptionsarepresentedtothesamebeesequally.Tocomparetheeffectivenessofdifferentnestingmaterialscommonlyusedbynativebeekeepers(namelyreedsandtrays)Iwillobserveinstalledspringmasonbees,Osmiaspp.,andcomparethenumberofviablebeecocoonscollectedfromeachnestingtypeattheendofanactiveseason.Oneachofthetwolocations,threegroupswillbeestablished:GroupA:OnlyReedsGroupB:OnlyTraysGroupC:ReedsandTraysOnSite1,SpringMasonBeecolonieswillbeestablishedonapropertywestofGregoryGardensat3290E9MileRoadinSaultSte.Marie,MI.Groupswillbeplacednolessthan200yardsfromoneanothertopreventinteractionbetweencolonies.Inthespringandsummer,thepropertywillhaveadequatefoodsourcesfortheinstalledbees,includingcrabappletrees,clover,andmiscellaneouswildflowers.Thereisalsoacreekforawater/mudsource.ThelocationforSite2islocatedat3588SRiversideDr,SaultSte.Marie,MIneartheSaintMary’sRiver.Onbothsites,additional“masonbeemud”willalsobeprovidednearthedomiciles,asmasonbeesrelyonclayeymudtoconstructbroodcells.InvataBeeMasonBeeAttractantwillalsobesprayedoneachdomicileattheperiodofinstallationandwhennestingmaterialsareadded;InvataBeeMasonBeeAttractantisaUSDA-patentedformuladesignedtomimicnaturalpheromonestoencouragenesting.Inaddition,birdguardswillbeplacedonalldomicilestopreventpredationofmasonbeelarvae.Inthefall,cocoonswillbeharvested,counted,andsortedbysite,colony,andnestingmaterialtypecollectedfrom,foratotalofeightbeebagslabeled1A,1B,1CR,1CT,2A,2B,2CR,and2CT,withtheinitialnumberindicatingwhethercocoonswerecollectedfromSite1orSite2andthefollowinglettersindicatingcolonyandnestingmaterialcollectedfrom.Afterthecocoonshavebeencountedbymeon-site,theywillbesentbymailtoCrownBeesHeadquartersinWoodinville,WA,toberecounted,testedforviability,andultimatelyredistributedtonativebeekeepersthroughoutthenationinthefollowingseason.CrownBeeswillsendmeaprofessionalbreakdownofthenumberofviablebeecocoonsfoundineachbag,whichmaydifferfromthetotalnumberofcocoonsfoundineachcolonyandnestingmaterialduetodisease,harvestingerrors,andotherfactors.BeeBagGroupingKey1A:CocoonsharvestedfromtheSite1OnlyReedgroup1B:CocoonsharvestedfromtheSite1OnlyTrayGroup1CR:CocoonsharvestedfromtheSite1ReedsandTraysgroup,takenfromtheReeds1CT:CocoonsharvestedfromtheSite1ReedsandTraysgroup,takenfromthetrays2A:CocoonsharvestedfromtheSite2OnlyReedgroup2B:CocoonsharvestedfromtheSite2OnlyTrayGroup2CR:CocoonsharvestedfromtheSite2ReedsandTraysgroup,takenfromtheReeds​​2CT:CocoonsharvestedfromtheSite2ReedsandTraysgroup,takenfromthetraysAstatisticalanalysistestwillbeusedtodeterminewhetherthedifferenceinviablebeecocoonnumbersissignificant(inotherwords,ifvariabilityincocooncountisduetochanceornestingmaterialtype).Ifthedifferenceisfoundtobesignificant,itmayindicateamoreeffectivenestingmaterial,“effective”indicatingthesuccessfulpropagationofmasonbeesasshownbythehighernumberofbeecocoons.Thesecondaryhypothesisofthisstudyexaminesbeepreferencefornestingmaterial.Todeterminebeepreference,specialattentionwillbepaidtotheCgroups,whichcontainbothreedandtraynestingoptions,duringweeklyobservations.Iwillnoteanddocumentbeeactivity,indicatingwhichnestingmaterialbeesareinvestigatingthemostandwhichnestingmaterialappearstobefilledthequickest,particularlyatthebeginningoftheseason.Inaddition,statisticalanalysistestswillcomparecocooncountsin1CRvs.1CT,2CRvs.2CT,1CRvs.2CR,and1CTvs.2CTtoexaminewhetherpreferencecanbedecipheredfromcocooncount.Becausepreferencedoesnotnecessarilyequatetoeffectivenesslevel,onlythenumberofcocoonswillbeconsideredinstatisticaltestsregardingpreference.Viabilityofcocoonswillnotbeconsidered.

SamuelMitchell
RelationsBetweenSeasonandDischargefromaHydroelectricReservoirontheDensityandTransportDistanceofZooplanktonintheSaintJosephRiverinBerrienCounty

Rivers,bytheirverynature,areaconstantlychangingecosystem.Riverprocessescanbefurthercomplicatedifareservoirisbuiltontheriver,withvariabledischargeenteringtheriversystemdownstream.Forexample,waterflowingthroughareservoir,anddownstreamfromthedam,canbevariablebyseason(e.g.,wetterversusdryermonths)andstormevents,whichcouldincreasetheoutflowofwatertoariverfromadam.Thesedynamicscansubsequentlyimpacttheecosystemandalterthecompositionandabundanceofcertainorganismsinarivernetwork.Onesuchimportantorganismgroupthatcanbeaffectedbyadamandvariablewaterflowarezooplankton.Zooplanktonarepartofthefoodchainofaquaticecosystemstheyprovideanimportantsourceoffoodforotherlargerorganismssuchasfishandcanaffectwaterquality.Zooplanktoncommunitiescanbedrasticallyaffectedinriversfromseasonaldynamicsmitigatedthroughupstreamdamsandreservoirs(Haveletal.2009).Althoughzooplanktonarecommonlystudiedinslowermovinglakes,zooplanktoncanalsoinhabitriversystemsandbetransporteddownstreamtootheraquaticecosystems(Portinhoetal.2016).Somezooplanktonspeciesalsoprefertoinhabitdifferentdepthsofriversandcanchangetheirdistributiondayversusnight.However,zooplanktonremainunderstudiedinriversversuslakeecosystems,andmoreinformationisneededtobetterunderstandhowseasonalandotherenvironmentalfactors(suchasstorms,whichareincreasinginfrequencyandintensityinmanyregionsbecauseofclimatechange)affectzooplanktonabundanceandtransportdownstreamfromreservoirs.MyprojectaimstostudytheeffectsofhydroelectricdamsonthedensityandtransportofzooplanktonintheSaintJoeRiverinBerrienCounty.Theeffectsofstormsarealsoanareaofinterestforthestudyastheycanincreasewaterdischarge,therebyaffectingthequantityandcompositionofzooplanktondownstreamfromwhatispresentinthereservoir.ThisriveralsoentersintoLakeMichigan,andsotheamountofzooplanktonenteringLakeMichiganfromanupstreamreservoircanbequantified.SampleswillbecollectedfromtheSaintJosephRiverusinga60µmmeshsize,50cmdiameter,zooplanktontownet.Zooplanktonwillbecollectedbyahorizontaltowinthreelocationsintheriverateachsamplingevent,thetownetwillbeadministeredhorizontallyfromakayakfor5minutestoallowenoughvolumeofwatertoentertocaptureenoughzooplanktontoquantify.Thesethreeriverlocationswillbe:justdownstreamofthereservoir,approximatelyequaldistancebetweenthereservoirandthemouthofLakeMichigan,andclosetothemouthofLakeMichigan.Inthereservoir(LakeChapin),averticalnettowwillcollectzooplanktoninadeepsectionofthereservoir;theseverticaltowswillcollectzooplanktonfrom1mfromthebottomtothesurface.SampleswillbecollectedonceamonthfromMaythroughAugustandbeforeandafterstormevents(3pairedbeforeandafterstormeventswilltargetedtobecollected).Afterthesamplesarecollected,thezooplanktonwillbepreservedusing99%isopropylalcohol,andwillbeanalyzedusingadissectingmicroscopeintheLSSUlabstoidentifyeachspecimentothegenuslevel.Specimenswillalsobemeasured,anddensityandbiomasswillbecalculatedforeachsample(Bumetal.1996).

MatthewHanson
EffectofPoly-D-LysineCoatingonmRNATransfectionEfficiencyinHEK293TCellCultures

Theabilitytomakeacellproduceaforeignprotein(fromanotherorganism)isanimportanttoolofcellandmolecularbiologyresearchwithmanyapplicationsinmedicine.Inageneralsense,someformofnucleicacid,DNAorRNA,needstoenterthecelltoprovidethegeneticcodefortheforeignproteintobeproduced.Thisstrategyforproteinproductioninatesttubeisthebasisforproteinsubunitvaccinesandrecombinantproteindrugssuchasinsulin.Therearevariouswaystocauseacelltoproduceaforeignprotein.Whennotusingavirus,thisprocessisknownastransfection.OnewaytoaccomplishthisiswithmRNAandvariousdeliverysystems.JustasthePfizer/ModernamRNAvaccinescauseyourhumancellstoproducethecoronavirusspikeprotein,thistechnologycanbeusedtocauseculturesoflivingcellstoproduceanyproteinforwhichthegeneticsequenceisknown(withlimitations.)Comparedtoculturesofbacteriaoryeast,mammaliancellculturesrequireveryspecificconditionstosurvive.Thisincludes:aspecificliquidnutrientmedium,temperature,gasmixture,andmanyrequireasurfacetogrowupon.Themostcommonmaterialthattissueculturevesselsaremadeofispolystyrene,aplastic.Naturallywaterrepelling,thesurfaceofpolystyrenetissuecultureplatesaretreatedwithoxygenplasmatomodifythesurface.Thisresultsinanegativelychargedmorewaterlovingsurfacethatallowsmanycelltypestoadhere.Somecellsrequire,andothersmaybenefitfrom,theadditionofanextracellular-matrixmimickingcoating.Thisisbecausecellsinmammalsareusuallysurroundedbyamatrixoffibers,likecollagen.Coatingsavailableinclude:thepolymerpoly-lysineandthevariousanimalproteinssuchascollagen,laminin,andfibronectin.Developedtomorecloselymimictheextracellularenvironmentofalivingorganism,thecoatingsincreasetheabilityofsomecelltypestoadheretothevessel(Corning,productmanual).Theaimofthisexperimentistoassesstheeffectthatapoly-D-lysinecoatinghasontheefficiencyofmRNAtransfectioninHEK293T(humanembryonickidney)cells.In1977thiscelllinewasproducedfromhumanembryonickidneytissue,andhasbeenacommoncelllineforproductionofrecombinantproteinsandhumanviruses.Thehypothesisisthatthedifferencethecoatingpresentstotheextracellularenvironmentcouldcauseadifferenceinthedevelopmentofthecellularmembrane.ThecellularmembraneisthebarrierthatmustbeovercomeinorderforthemRNAtoenterthecellandbetranslatedtomakeaprotein.Therefore,differencesinmembranecompositionorstructurecouldcausedifferencesintheabilityofmRNAtoenterthecellandbetranslatedintoprotein.ThepercentageofcellsthatthemRNAsuccessfullyentersandproducestheproteinisknownastransfectionefficiency.Theforeignproteinbeingchosenisgreenfluorescentprotein.UnderUVlight,greenfluorescentproteinemitsagreenlight.Thisuniquepropertyoffluorescentproteinsallowsforeasyidentification,becausehumancellsdonotproduceanytypeoffluorescentproteins.Tissuecultureplatesandtissuecultureplatescoatedwithpoly-D-lysinewillbeseededwiththesamestockofHEK293Tcells.Thecellswillbegrownandhandledundertheexactsameconditionsandwilldifferonlyintheadditionofapoly-d-lysinecoating.Whilegrowing,thecellswillbeobservedviamicroscopetoobserveanydifferenceintheappearanceofthecellmembraneorothergrowthcharacteristics.OncethecellshavegrownenoughtheywillbetreatedwithmRNAdeliveredbythecommontransfectionreagentlipofectamine.ThetransfectionefficiencywillbedeterminedusingtheThermoFisherCountessautomatedcellcounter.Ithastheabilitytocountthetotalnumberofcellsandthenumberofcellsexpressingthegreenfluorescentprotein.Givenasapercentagethisrepresentsthetransfectionefficiency.Meantransfectionefficiencyofcoatedwellswillbecomparedtothenon-coatedwellsviaanappropriatestatisticaltest.

MichelleGlasscock
SuicidalBehaviors,ACEs,andEducationalAttainment: APotentialBasisforDeveloping MoreEffectiveAssessmentToolsforEducationProfessionals

Thebrainundergoesrapiddevelopmentduringchildhood.Experience,bothpositiveandnegative,playsanimportantroleinthisprocess.Whenchildrenexperienceabuseand/orneglect,itcaninterferewiththeirabilitytobesuccessfulinschool.Thisputsthematriskofnotreachingtheirfullpotentialacademically,limitingoccupationalchoices,andultimately,earningpotential.Researchhasalsoshowncorrelationswithnumerousharmfuleffectsinadulthood(e.g.,physicalandmentalhealthproblems;CDC,2019)aswellasincreasedriskofsuicideandsubstanceabuse(Park,etal.,2021;Whitesell,etal.,2009).Again,theimpacttoqualityoflifecanbedevastatingwhenthesechildrenreachadulthood.Therefore,earlyidentificationandinterventioniscrucial.Currently,K-12schoolsusetheAdverseChildhoodExperiences(ACEs)questionnaireasascreenertoidentifystudentsatriskforpooracademicachievement(Catterfeld,2019;NEA;Pataky,etal.,2019).However,thisisonlyoneareaofconcernrelatedtochildmaltreatment.Childrens’suicidalbehaviorsandinitialuseofsubstancesoftenbeginbeforereachingadulthood(CDC,2019;HHS;2019).ThisraisesthequestionofwhethertheuseoftheACEsshouldbeexpandedtoscreenforriskofsuicideandsubstanceusetoprovideamorecomprehensiveinterventionplan.However,afewissuesneedtobeexplored.First,therelationshipbetweenACEsandsuicidehavebeeninvestigatedwithanemphasisonsuicideattemptsandcompletionsbutnotsuicidalbehaviors.Second,asimilarresearchfocusexistsbetweeneducationalattainmentandsuicidalbehaviors(i.e.,educationalattainmenthasbeenlinkedtosuicideattempts/completionsbutnotbehaviors).Therefore,thepurposeofthisresearchistoestablishalinkbetweensuicidalbehaviors,ACEs,andeducationalattainmentthatwouldpermitearlieridentificationofriskbeforeasuicideattemptorcompletionoccurand/oreducationalattainmentisthreatened.Toexplorethisquestion,asampleof150adultparticipantsrecruitedthroughProlificwillbeassessedforACEs,suicidalbehaviors,educationalattainment,andsubstancemisuse.ChildhoodmaltreatmentwillbemeasuredusingtheACEs,suicidalbehaviorswillbeexaminedwiththeSuicidalBehaviorsQuestionnaire–Revised(SBQ-R),aquestionwillaskparticipantswhattheirhighestlevelofeducation,andtheNIDA-ModifiedASSIST(NMASSIST)willbeusedtocharacterizesubstancemisuse.Ifacorrelationisfoundbetweenthesevariables,itwouldlendsupporttoexpandingtheACEsscreenertoincludesuicidalbehaviors,givingeducationalprofessionalsbettertoolswithwhichtohelpstudentssucceed.

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Fall2021GrantAwardees

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OliviaHohman
TheBioremediationofPolyethylene,Polyurethane,PolypropyleneandPolystyrenebyAspergillusNigerandBacillusSubtilis

Asthemanufacturingandconsumptionofplasticshascontinuedtoincreaseovertheyears,plasticdebrisposesahugethreattomanyecosystems.Theamountofplasticaccumulatingintheenvironmenthasbeensteadilyincreasingasaresultofplasticdurabilityandlightweightnature(1).Anestimated300milliontonsofplasticareproducedandusedyearly(1).Plasticsareman-madematerialsmanufacturedfrompolymersorlongchainsofrepeatingmoleculesderivedfromoil,naturalgas,andplantssuchascornandsugarcane.Inadditiontoreplacingsteelincars,andwoodinfurniture,plasticshavereplacedpaperandothercellulose-basedproductsforpackaging.Biodegradationoftheseabundantplasticsmaybethekeytocontrollingthisproblem(1).Amoreefficienttechniqueofplasticdegradationmaybeachieved,byunderstandingthemechanismofpolymerdegradation.Thisrequiresresearchersbecomefamiliarwithhowcompoundsaremetabolizedbyexistingorganisms,aswellasidentifyingneworganismscapableofbiodegradationandcharacterizingtheirmetabolicabilities(2).Biodegradationisacomplexnaturalprocesstodecomposesubstancesandinvolvesseveralsteps:1)fragmentationofmaterialsintosmallerfunctions(bio-deterioration);2)secretionoffreeenzymesandfreeradicalspeciestocleaveoligomers,dimersandmonomers(depolymerization),3)recognitionsandreorganizationofselectmolecules(assimilation),and4)oxidationofsimplemolecules,andsaltsfrommetabolites(mineralization)(3).Whileanumberofbacteriahavebeenfoundtobeexcellentatbiodegradationrelativelyfewfungihavebeenidentifiedascapableofplasticdecomposition.Polyethylene,polyurethane,andpolystyrenearethepredominanttypesofplasticsusedinindustryandmanufacturing,andallhaveshownsusceptibilitytobiodegradation(2).Plasticsarepolymersgeneratedbythecondensationofpolyisocyanateresultinginacarbonpolymercomposedofurethanelinkages(2).Alterationsinthespacingbetweenurethanelinkages,aswellatthesubstitutions,canchangethepropertiesoftheresultingpolymerfromlinearandrigidtobranchedandflexible(2).Microorganismsinprolongedcontactwithplasticwasteshavebeenfoundtoadaptandmaximizetheirdegradationpotential(4).Thesemicroorganismsare“reportedtoproducesomespecialenzymesviz.intracellularandextracellular,whichenablethemicrobestodisintegratethepolymerintoseveralmonomersanddimers,whicharebeingusedbythemicrobesasacarbonsource”(7).Inthisstudy,specialinterestwillbepaidtoAspergillusniger,andBacillussubtilisfortheirabilityasknowndegraders.Baselinegrowthandbioremediationpotentialoftheseorganismsbothontheirownandinconsortiawillbeobservedagainstavarietyofplasticsubstrates(polyethylene,polyurethane,polypropylene,andpolystyrene).Activityofthemicroorganismsagainsttheplasticsubstrateswillbeobservedinthelaboratorysetting.Understandingthedetailedmetabolismoftheorganismswhichnaturallydegradeplasticsmayshedlightonthechallengesofremovingplasticwastetopreserveourenvironment.GrowthEstablishmentA6mLstartercultureofBacillussubtiliswillbepreparedusingnutrientbrothandplacedintheshakingincubatorfor24hrsat30℃and220rpm.100μLofthestarterculturewillbespreadonanutrientagarplateandincubatedfor24hrsat30℃.Twosinglecolonieswillthenbepickedfromtheplateandaseparateglycerolstockwillbemadeforeachcolony.Aspergillusnigerwillberehydratedusing6mLsteriledistilledwaterandallowedtositfor24hrs.100μLofthestarterculturewillbespreadonaSabouraudDextroseAgar(SDA)alongwith100mg/mLampicillinplateandincubatedat37℃untilgrowthisobserved,2weeks,forfungi(2.3.5).Twosinglecolonieswillthenbepickedfromtheplateandaseparateglycerolstockwillbemadeforeachcolony.BioremediationTestsSheetsofpolyethyleneandpolyurethane,andshavingsofpolypropyleneandpolystyrenewillberinsedwithsterilewater,dried,weighedandsterilizedwith70%Ethanol(4).EstablishmentofbaselinebioremediationpropertieswillbedonewithnutrientbrothforBacillussubtilisandSabouraudDextrosebrothwithampicillinforAspergillusniger.1gofpolyethylene,polyurethane,polypropyleneorpolystyrenewillbeaddedtothecultureandgrowthwillbecarriedoutat30℃forBacillussubtilisand37℃forAspergillusnigerforuptosixweeks.Theplasticsheetorshavingswillberemoved,cleanedwith70%ethanolanddeionizedwater,driedintheovenforonehour,andweightedevery2weeks.Onceweightedthesheetorshavingswillbeaddedbacktotheculture.Fromtherethedegradationcanbemeasuredbytheweightreductionofthedrysamples.Threereplicateswillbedoneofpolyethylene,polyurethane,polypropylene,andpolystyrenewithAspergillusNiger,andBacillusSubtilis.Inadditiontothenegativecontrol,whichwillcontainonlyampicillininsyntheticmedia,apositivecontrolwhichcontainseachtypeofplasticwithampicillininsyntheticmediawillbeprepared.Thecontrolsshouldshownofungalorbacterialgrowthandnoweightreductionduetodegradation.BioremediationbyfungiwillbecarriedoutinsyntheticCzapekDoxbroth(pH6.8,100mg/mLampicillin)(6).BioremediationbybacteriawillbecarriedoutinDavisbroth,aminimalingredientmedium(6).1gofpolyethylene,polyurethane,polypropyleneorpolystyrenewillbeaddedtothecultureandgrowthwillbecarriedoutat30℃forBacillussubtilisand37℃forAspergillusnigerforuptosixweeks.Theplasticsheetorshavingswillberemoved,cleanedwith70%ethanolanddeionizedwater,driedintheovenforonehour,andweightedevery2weeks.Onceweightedthesheetorshavingswillbeaddedbacktotheculture.Fromtherethedegradationcanbemeasuredbytheweightreductionofthedrysamples.Threereplicateswillbedoneofpolyethylene,polyurethane,polypropylene,andpolystyrenewithAspergillusNiger,andBacillusSubtilis.Inadditiontothenegativecontrol,whichwillcontainonlyampicillininsyntheticmedia,apositivecontrolwhichcontainseachtypeofplasticwithampicillininsyntheticmediawillbeprepared.Thecontrolsshouldshownofungalorbacterialgrowthandnoweightreductionduetodegradation.

MichaelHillary
ComparingLitterDecompositionAcrossaGradientofWetlandHealth

Background:CoastalwetlandsareavitalpartoftheGreatLakes(Uzarkietal.2019),byprovidinghabitat,nutrients,fisheriesandbiodiversitytotheecosystem(Wetzel1992,Brazneretal.2000andWeietal.2004).Manyspeciesrelyonthesewetlandsforcrucialnurseryhabitat,suchastheyellowperch(Parkeretal.2012).Inadditiontobenefitingtheecosystem,coastalwetlandshavebeenestimatedtoproduce69billionUSDannuallytotheeconomysurroundingtheGreatLakes(Krantzbergetal.2008).Inthepast100years,humanactivitieshavecausedadepletionofwetlandhabitatbyover50%(Kriegeretal1992)andtheseactivitiescontinuetothreatentheremainingwetlands(Trebitzetal.2007).Forthepastdecade,theUSEPA’sGreatLakesmonitoringprogramhasbeenassessingwetlandhealth.Themonitoringprogramhasbaseditscategoriesofwetlandhealthlargelyonwhatspeciescanorcannotbefoundthere(Uzarkietal.2019).Thisonlyaccountsforthestructureofthewetlandanddoesn’trepresenttheentireworkingsofitsecosystem.Afunctionalindicatorcanaidindetermininghowitisworkingorhowhealthyitisbasedonitsfunction(Sandinetal.2009).Onecommonwaytotestanecosystem’sfunctionalhealth,istomeasuretherateofplantdecomposition(Youngetal.2008,Udyetal.2006andLambertietal.2017)becausethisfunctionisaffectedbywaterqualityandbacterialcommunities.Objective:Theobjectiveofmyseniorresearchistoassesswhethercurrentcategoriesofwetlandhealthcorrespondwiththeecosystem’sfunctionalhealth.Thiswillbedoneusingdecompositionacrossagradientofwetlandhealth.Asanalternativemeasurement,thenumberofbacteriafeedingontheplantmatterwillbeassessed.Ihypothesizethattherateoflitterdecompositionwillbequickerinhigherqualitywetlandscomparedtolowerquality,andthatmicrobialrespirationwillbehigherinhighqualitywetlandscomparedtolowquality.OverviewofMethods:Litterdecompositionwillbemeasuredin12wetlandsduringthesummer,with3categoriesofhealth:high,mediumandlowacrosstheSt.Mary’sRiver,locatedintheEasternUpperPeninsulaofMichigan.HealthcategorieswillbedeterminedusingvaluesassignedbytheGreatLakesCoastalWetlandMonitoringprogrambasedonmacroinvertebratesandfishes.Cattailswillbeusedastheplantmaterialasitiscommonlyfoundinthecoastalwetlands.Atleast156meshbagswillbeconstructedfrom1.27mmmeshsheets:20x20cm(Benfield1996),andeachsitewillcontain13bags.Tengramsofdriedcattailwillbeweighedandplacedineachmeshbag(Lambertietal.2017).Bagswillbelabelledwithplastictags,sealedwithavacuumsealerandtiedontorebarusingbraidedfishingline.(Andersonetal.2002).Sixretrievalperiodswilltakeplaceeveryfewweeksbeforethewaterfreezesover.Threemoreretrievalperiodswillbecompletedaftertheicemeltsnextspring.Oncemeshbagsareretrieved,theywillbedried,weighed,ashed,andre-weighedtodeterminelossoforganicmaterial(i.e.,decomposition).Therateofmasslossovertimewillbeusedtocalculatedecompositionrate.Onthe6thretrieval,thebacterialcommunityontheplantmaterialwillbemeasuredforeachwetland.Thiswillbedonebymeasuringtherisesanddropsinoxygenduringperiodsoflightanddarkincubation.Duringthelight,photosynthesiswilloccur,andoxygenlevelsrise.Duringthedark,respirationwilloccur,andoxygenlevelsfall.IncubationwilltakeplaceinAshmunBaytoregulatetemperatureandsunlightthefollowingmorningofretrieval.Darktoteswillbeplacedoverlighttotesandthenlitterbagswillbeplacedwithinthetotes.Thiswillallowthebagstobeexposedtolightanddarkconditionswithoutmovingthebags.Oneofthetotesetswillnotcontainanymeshbagsandwillserveasacontrolofmicrobialrespirationwithoutthelitter.DO(dissolvedoxygen)loggerswillbeplacedineachshoeboxtotetomeasureDOchangesevery30minutes.Changesindissolvedoxygenduringthe“dark”periodand“light”periodwillbeusedtocalculaterespirationratesforeachtreatment.AnANCOVAtestwillbeusedtodifferentiatethelossofmassamongwetlandtypesovertime.Themasslosswouldbethedependentvariable,wetlandtypeistheindependentvariable,andtimeisthecovariate.Thebacterialcommunitygrossprimaryproduction(GPP)willbecalculatedaddingtheoxygenchangeduringthedark(R)bytheoxygenchangeduringthelight(NPP).RespirationdifferencesacrosswetlandtypeandtimewillalsobetestedusinganANCOVA.

PaxtonSpencer
Macroalgae inSaltwaterAquaponicSystemsReduceNutrientsinEffluent

Saltwateraquaponicsisarelativelynewareaofstudyasamajorityofsystemsarefreshwater.Howeverwithgrowingconcernoverthelackoffreshwaterinmanycountriesthissetupisbecomingthenewwaytofarminanefforttoconservedrinkingwaterandbringfoodtotheseareas.Aquaponicsisknowntoconserveover90%ofwaterandbethesustainablewaytoraisetwofoodsourcesthatworktogether.However,effluentfromthesesystemscancauseeutrophicationinwatershedsaswellasincreasethesalinityofthesoilwhennotproperlyhandled.RecentpublicationsasofthisyearbegantolookatIMTAsorintegratedmulti-trophicaquaculturesystemsasamodelinreducingthenutrientloadofeffluent.Theideaofthissystemistohavethreelayersfromaprimaryproducerallthewaytoaprimaryconsumerinanefforttoreducethesolidsintoworkableenergyformacroalgae.IMTAsystemshavebeenstudiedinopenoceansettingswheretheyhavefoundthatthewastewithintheround-penssignificantlydecreasedcomparedtopenswithout.Theyalsofoundthatthemacroalgaegrewmuchfasterinthissettingthanitdidonitsowninsimilargrowthpenswithoutfish.Therefore,takingthismulti-levelsystemtolandbasedshrimpfarmscouldsolvetheirexcessnutrienteffluentproblemwhileinturncreatingmultipleavenuesforbusiness.TorepresenttheaquaponicsystemsetupeachaquariumwillhaveSalicorniaeuropaea,aplantthatwillaidinnutrienttestingaswell.Thesespeciesofplantsdonotlikeexcessnutrientsandhavefoundtogrowwhentherewasnotanabundanceofphosphateornitrogeninthewater.Therefore,theirgrowthratewillbeanothertestoutsideofbasicchemistrytestingtoseehowmuchnutrientsareinthewater.Thishalophyte,orsaltmarshplant,isjustasmuchofadelicacyasnoriseaweedisintheculinaryworld.Overall,findingthatmacroalgaecanreducenutrientsinsaltwateraquaponicsystemstherewillbepossibilitiesofexpandingthefieldandreducingeffluentwaste.

ReyannaRutledge
PrevalenceofCaspase-9MutationsintheLungCancerPopulationoftheEasternUpperPeninsula

“Cancerisknownforbeingtheleadingcauseofdeathsworldwideannually.Whilebeingthesecondmostcommondiagnosedcancer,“lungcanceristheleadingcauseofdeathsworldwide”(MayoClinic,2021).Lungcancerisatypeofabnormalcellgrowththatoccursinthelungs.Toputintoperspective,almosttenmillioncancerpatientsdiedin2020,whilenearlytwomillionofthosedeathswerecausedbylungcancer(Sung,H.,etal.,2021).Therefore,identifyingdifferentcausestolungcancershouldbeprioritized.Todayweknowtherearevastamountsofcarcinogensintheworld.Carcinogensareenvironmentalfactorsthatcausecancer.Oneexampleofthiswouldbecigarettes.Infact,smokingisthebest-knowncauseoflungcancer.Ontheotherhand,lungcancercanalsooccurinpatients’whohaveneversmoked.Thisleadstomorebackgroundresearchsuchas;genetichistoryandotherenvironmentalfactorstoidentifycauses.Canceristhedevelopmentoftumorscausedbyrapid,uncontrolledproliferationofcellsinthebody.Whatcausesthesecellstorapidlyproliferate?Thedevelopmentofcancer,isconsideredamultiplestepprocessatthecellularlevel(Cooper,1970).Mutationsarethefirststepleadingintoanycancerdevelopment.Thesemutationsaffectcelldivisionbyalteringhowmanyproteinsarepresentandtheirfunctionofregulatinghowthecellsgrow,divideandhowtheproteinrepairsDNA(Gale,2020).OncethemutationalterstheDNA,continualreplicationofthealteredDNAwilloccur.Thiscanleadtomasscelldivisionoftheabnormalcellscausingtumors.Onetypeofmutationcausesaproto-oncogenetoconverttoanoncogene.Proto-oncogenespromotecellgrowthandthedifferentiationofcells.Thiscanbeviewedascellactivity.Whenaproto-oncogeneismutatedintoanoncogeneitcanbeover-activeleadingtoexcessivecellproliferation(Gale,2020).Anothertypeofmutationcausesinactivationoftumorsuppressorgenes.Thehealthytumorsuppressorgenesarepartofthecellularbalancethatpromotesapoptosisandinhibitscelldivision.Balancingthecellsgrowthanddeathmaintainsahealthybody.Thealterationofapoptosiscausedbyoncogenesandmutatedtumorsuppressorgenesisthefocusofthisstudy.Apoptosisisprogrammedcelldeath,andthealterationstoapoptosisarehighlyrecognizedinaffectstohumancancers(OlssonandZhivotovsky,2011).Thisformofcelldeathiscontrolledbycaspasesandotherregulatoryfactors.Thereareaseriesofcaspasegenes,orderedbynumberandcategorizedintogroupsbasedofffunction.ThefocusofthisstudywillbetowardsgroupII,sinceitisresponsibleforapoptosis.GroupIIisthensplitintotwoclasses:“initiator(apical)caspases(caspase-2,-8,-9and-10)andeffector(executioner)caspases(caspase-3,-6and7)”(OlssonandZhivotovsky,2011).Inshort,theinitiatorcaspasesendsoffanenzymethatsignalstheeffectorcaspasetotargetspecificproteinsoftheunhealthycelltoinitiateapoptosis.Tospecifyforthisresearch,thefocuswillbeontheinitiatorcaspase,caspase-9,andpolymorphismsorgenealterationsthatoccurtoit.Thereareseveralpolymorphismsormutationsthatoccurinthecaspasegenes.Alterationsofthecaspasegeneshavebeencorrelatedtocausingmultipletypesofcanceraswellaslungcancer.Therefore,findingspecificmutationslinkedtospecificcancersisdifficult.Instudiesspecifiedtocaspase-9genealterations,thereisalotofresearchonpolymorphismsrs4645978andrs4645981andtheirlinktolungcancer.Afewstudieshaveshownapossiblecorrelation,morespecificallyofthers4645981polymorphismtolungcancer.Forexample,onestudysuggestsapossiblecorrelationtothedevelopmentoflungcancer(Park,J.,etal.,2006).Inanotherstudy,threepolymorphismsofdifferentcaspasegeneswerelookedat,andwereassociatedtowiththeriskoflungcancer(Lee,S.Y.,etal.,2010).Thepossiblelinkbetweenthecaspase-9polymorphismsandlungcancer,couldbestepstowardfindingtheothercausesoflungcancer.Therefore,thefocusofthisresearchwillbeonfindingmoredataofpolymorphismsonthecaspase-9geneinapopulationthatcouldpossiblybecorrelatedtolungcancer.Methods:​ThisstudywillbeconcentratedtotheEasternUpperPeninsulapopulation,withparticipationoftwentylungcancerpatientsandtwentynon-lungcancerpatientsasthecontrol.LungcancerpatientswillbeidentifiedbyWarMemorialHospitalOncologydepartment,andaskediftheyareinterestedinparticipatinginthestudy.Awaiverwillbegiventoeachparticipant,explainingthestudyandnotingthatidentityofparticipantswillnotbetracedfromDNAsample.Eachsignedwaiverwillhaveacodenumberthatmatcheswiththenumberonagivenswab,tokeepidentitiesanonymous.Theparticipantswillswabtheinsideoftheircheekandplaceswabbackintowrapping,forlatertesting.TheswabswillthenbetakentotheLSSUlabwhereDNAextractionandamplificationviaPCRwilltakeplace.​DNAamplificationofthecaspase-9genewillbeperformed,usingpolymerasechainreaction(PCR).PCRusingspecificprimerstomakecopiesoftargetedregionsofthesubjectDNA.EachcycledoublestheamountofDNAcopied.After25-30cyclestherecouldbe@onebillioncopies.Thiscanberepeatedforseveralduplicationsofthesamesegment(NationalHumanGenomeResearchInstitute,2020).Followingpreviousstudies,thePCRproductswillbesentouttoPsomagenInc.tosequencetheDNA,tolookatseveralmutationsinsteadofjustthetwomentioned(Lee,S.Y.,etal.,2010).Duetopricing,sendingsamplestobesequencedisconsideredthebestoptionforanalysis.​AftertheDNAsequencingiscompleted,thebasesequencecanbeanalyzed.EachgroupwillbeanalyzedthroughapairedTtest.Thiswillcomparetheprevalenceofrs4645978andrs4645981thatoccurredinthecontrolgrouptotheprevalenceinthelungcancerpatients.Oncetheprevalencecanbecompared,thetotalmutationoccurrencecanbecalculatedforthepopulationstudiedintheEasternUpperPeninsula.

HannahBrood/TeamAMORE
LSSURobotX

ThepurposeofthisresearchprojectistodesignanddevelopanUnmannedSurfaceVehicle(USV)mobilerobotthatiscapableofcompletingtheautonomoustasksspecifiedbythe2022MaritimeRobotXChallenge.ThiscompetitionisopentoundergraduateandgraduatestudentteamsandwillbeheldinSydney,NewSouthWales,Australia.Inordertocompletetheserequiredtasks,theUSVwillneedtobeabletoautonomouslyidentifyandlocalizedifferentmaritimeobjects,followpaths,holdpositions,andcommunicatedatatolandstationsandanothermobilerobot.TheUSVmustalsobeabletocorrectitselfinthepresenceofwindandwaves.TheRobotXcompetitiondirectlyadvancesthefieldofautonomousmaritimeoperationssincethesetaskrequirementsareactiveresearchtopicsinfieldssuchasoceanengineeringandmaritimemobilerobotics.Thisisimportantsinceoceansandseasarehighlydynamicenvironmentsthataccountfor71percentoftheEarth’ssurfacearea.AdvancingmaritimeautonomywillultimatelyleadtobettermappingofEarth’soceans,improveddatacollection,anddevelopmentofautonomouspassengerandfreighttransportation.Inordertocompletethisproject,aWaveAdaptiveModularVessel(WAM-V)hasbeengrantedtoLSSURoboticstoserveasthebaseplatformfortheUSV.TheWAM-Visa16-footlong,catamaran-stylevesselwithapayloadcapacityof485pounds.Tomeettheprojectrequirements,theWAM-Vwillneedpropulsion,remotecontrol,emergencystop,communication,power,vision,acoustic,andguidance,navigation,andcontrol(GNC)systems.Thepropulsionsystemisusedtomovethevesselandwillconsistofmotorsandsteeringmechanisms.Theremotecontrol,emergencystop,andcommunicationsystemsareusedtomanuallycontrolandshutdownthevessel.Thepowersystemwillconsistofon-boardbatteriesthatwillprovidepowertotheothersystemsofthevessel.ThevisionandacousticsystemswillusetechnologysuchasLiDAR,cameras,andhydrophonestoprovideinformationaboutthesurroundingenvironmenttotheUSV,allowingittomakedecisionsonhowitshouldbehave.TheGNCsystemwillconsistofacomputerthatrunscontrolsoftwareandaprogrammablecircuitboardthatsendssignalstotheothersystemsoftheUSV.ThisresearchprojectisaSchoolofEngineeringandTechnologySeniorDesignProjectforTeamAMORE(AutonomousMaritimeOperationsandRoboticsEngineering).TeamAMOREhassixmembers. ThefacultyadvisorfortheteamisDr.EdoardoSarda.Asanontechnicalrequirementfortheproject,TeamAMOREhascreatedtheLSSURobotXClub.Thisprojectwillgivebothteamandclubmemberstheopportunitytobeinvolvedinamultidisciplinarymobileroboticsprojectthatinvolvesmarketing,workingwithindustryprofessionals,collaboratingwithinternationaluniversitystudents,andcompetinginaninternationalcompetition.

CatherineQuon
CompetitionBetweenLactobacillusAcidophilusandStaphylococcusAureusinEggSaladat27DegreesC

Staphylococcusaureusisthebacteriaresponsiblefor41%offoodpoisoningoutbreaks(Hernàndez-Cortezetal.2017).IntheUnitedStates,thereareapproximately241,000casesofStaphylococcalfoodpoisoningannually(Kadariya2014).S.aureusisaconcernasitistolerantofhighsaltandsugarconcentrationsandproducesheatresistanttoxins(Kataokaetal.2015,Huetal.2018).Onetosixhoursafteringestionofthetoxincausessymptomsincludingvomiting,diarrhea,fever,andcramping(Asao,2002,Huetal.2007,Kimetal.2011).PotlucksandpicnicfoodsareatriskforS.aureuscontaminationastheyareleftatambienttemperatureforhours,promotingthegrowthofbacteria.30%ofthehumanpopulationcarryS.aureusontheirbodiesandcanspreadthebacteriathroughphysicalcontactandrespiratorydroplets(Argudínetal.2010,Asaoetal.2003,Kimetal.2011).EggsaladisaclassicpicnicfoodthatissusceptibletoS.aureuscontamination.CompetitivemicrobialgrowthbetweenLactobacillusacidophilusandStaphylococcusaureusmayaffectthegrowthofcolonies.LactobacillusproduceshydrogenperoxideandlacticacidwhichhasbeenshowntosuppressthegrowthofS.aureus(DahiyaandSpeck1968,Sameshimaetal.1998).AsL.acidophilusiscommonlyfoundinyogurts,itcanbeeasilyincorporatedintotheeggsaladrecipe.TheobjectiveistodeterminetheeffectofLactobacillusacidophilusonthegrowthofStaphylococcusaureuscoloniesineggsaladat27°C.TrypticSoyBroth,NutrientAgar,LactobacilliMRSbroth,andLactobacilliMRSagarwillbepreparedaccordingtothemanufacturer’sinstructions.S.aureusATCC43300willbeobtainedfromLSSUfrozenbacterialstock.TheL.acidophilusfromtheprobioticcapsulewillbesuspendedinbroth.Gramstainingwillbedonetoconfirmtheidentityofthebacteriaafterstreakingforisolatedcolonies.Theeggsaladwillbepreparedaccordingtotherecipe.ThesamplebatchofeggsaladwillbeinoculatedwithbothS.aureusandL.acidophilus.ThecontrolbatchofeggsaladwillonlybeinoculatedwithS.aureus.Thetwobatcheswillbeincubatedat27°CforsixhourstoimitateasummerpicnicinSaultSte.Marie.Threesamplereplicateswillbetakenfrombothbatcheseveryhourandspreadonanagarplate,thenincubatedfor24hours.S.aureuscoloniesoneachplatewillbeidentifiedandcounted.Themeanandstandarddeviationwillbetakenforeachofthethreereplicates.Ascatterplotgraphfor“AverageCFUS.aureus/gvs.timeincubated(hrs)”willbemadetocomparethecontroltothesample(withL.acidophilus).Standarddeviationerrorbarswillbeincludedonthegraphforeachtimepoint.T-testswillbetakenateachtimepointtodetermineifthedifferenceinnumberofcoloniesofS.aureuspergramofeggsaladissignificantenoughtosaythatL.acidophilushadaneffect.Argudín,M.A.,M.C.MendozaandM.R.Rodicio.2010.FoodPoisoningandStaphylococcusaureusEnterotoxins.Toxins2010(2):1751-1773.Asao,T.,Y.Kumeda,T.Kawai,T.Shibata,H.Oda,K.Haruki,H.NakazawaandS.Kozaki.2002.Anextensiveoutbreakofstaphylococcalfoodpoisoningduetolow-fatmilkinJapan:estimationofenterotoxinAintheincriminatedmilkandpowderedskimmilk.EpidemiologyandInfection130(1):33-40.Dahiya,R.S.andM.L.Speck.1968.HydrogenPeroxideFormationbyLactobacilliandItsEffectonStaphylococcusaureus.JournalofDairyScience51(10):1568-1572.Hernàndez-Cortez,C.I.Galma-Martínez,L.U.Gonzalez-Avila,A.Guerror-Mandujano,R.C.SolísandG.Castro-Excarpulli.2017.FoodPoisoningCausedbyBacteria(FoodToxins).Pages33-72inPoisoning-FromSpecificToxicAgentstoNovelRapidandSimplifiedTechniquesforAnalysis.N.Malangu(Editor).IntechOpen.Hu.,D.-L.,G.Zhu,F.Mori,K.Omoe,M.Okada,K.Wakabayashi,S.Kaneko,K.ShinagawaandA.Nakane.2007.Staphylococcalenterotoxininducesemesisthroughincreasingserotoninreleaseinintestineanditisdownregulatedbycannabinoidreceptor1.CellularMicrobiology9(9):2267-2277.Hu,J.,L.Lin,M.ChenandW.Yan.2018.ModelingforPredictingtheTimetoDetectionofStaphylococcalEnterotoxinAinCookedChickenProduct.FrontiersinMicrobiology.9(1536):1-11.Kadariya,J.,T.C.SmithandD.Thapaliya.2014.StaphylococcusaureusandStaphylococcalFood-BorneDisease:AnOngoingChallengeinPublicHealth.BioMedResearchInternational2014(827965):1-9.Kataoka,A.,E.Enache,C.Napier,M.HaymanandL.Wedding.2016.EffectofStorageTemperatureontheOutgrowthandToxinProductionofStaphylococcusaureusinFreeze-ThawedPrecookedTunaMeat.JournalofFoodProtection79(4):620-627.Kim,N.H.,A.-R.YunandM.S.Rhee.2011.PrevalenceandclassificationoftoxigenicStaphylococcusaureusisolatedfromrefrigeratedready-to-eatfoods(sushi,kimbabandCaliforniarolls)inKorea.JournalofAppliedMicrobiology111:1456-1464.Sameshima,T.,C.Magome,K.Takeshita,K.Arihara,M.Itoh,andY.Kondo.1998.EffectofintestinalLactobacillusstarterculturesonthebehaviourofStaphylococcusaureusinfermentedsausage.InternationalJournalofFoodMicrobiology41(1998):1-7.

KatjaWilhelms
ComparingtheEffectsofTraditionalandE-CigaretteVaporonCandidaAlbicansGrowthandVirulenceFactorExpression

Candidaalbicansisaprevalentandpossiblypathogenicyeastfoundonmosthumanbodies(Kendrick,2017).Whileittypicallyactsasaninterdependentfungusthatcausesthebodynoharm,anovergrowthofC.albicansmayleadtoinfectionandseriousdiseasesknownascandidiasis.AsdefinedbytheCenterforDiseaseControl(CDC),candidiasis,alsoknownasoralthrush,isaninfectionintheoralcavitycausedbyaCandidaspecies.Symptomsincludewhitepatchesontheinnercheeks,tongue,roofofthemouthandthroat,rednessorsoreness,lossoftaste,andpainwhileeatingorswallowing(CDC,2021).C.albicansandotherCandidaspeciesarepresentintheoralcavityofupto75%ofthepopulation(Ruhnke,2002).Furthermore,ahighincidenceofC.albicanshasbeenfoundinhospitalsettings.Itwasthemostfrequentlyisolatedfungalpathogen(59.7%)inhospitalenvironments(Beck-SagueandJarvis,1993).C.albicanshasmanydifferentfactorswhichincreasethedegreebywhichitcancausedisease,makingitmorepathogenicthatotherCandidaspecies.Thesefactorsincludeitsattachmentstyle,abilitytochangeform,antifungaldrugresistance,abilitytocongregate,anditssecretionofaspartylproteinase(Sap).ThisprevalenceandwidespreadmakesthestudyofC.albicansanimportantfocusinthebiomedicalfield.Thisresearchprojectaimstofurtherexplorethesecretionofaspartylproteinase(Sap)asanincreasedpathogenicfactorinC.albicans.Sapisanextracellularenzymethatcontributestothenutrientacquisition,invasion,tissuedamage,evasionofhostresponseofC.albicans(Naglik,2003).Forexample,thesecretedproteinasefromC.albicansdecreasestheantimicrobialdefenseefficacyofhumansaliva(Kaminishi,1995),makingC.albicanstoughtokillintheoralcavity.Specifically,theSAP2geneisoneofthemostcommonlyexpressedgenesinpatientswithoralcandidiasis.Theintroductionoftraditionalcigarettesinthe1960’sfueledariseinthesmokingoftobacco.However,astimepassed,moreandmoreresearchofferedevidenceontheharmfullong-termhealtheffectsofcigarettesmoking.Ithasbeenlinkedtomultipleneurological,cardiovascular,andpulmonarydiseases,aswellashealthproblemssuchasasthmaduetosecond-handsmokeinchildren(Das,2003).Whiletherehasbeenadownwardtrendintheiruse,in2019theCDCestimated14.0%(34.1million)ofU.S.adultswerecurrentcigarettesmokers.Recently,electroniccigarettes(e-cigarettes),alsoknownasvapes,haveincreasedinpopularity.Theyareproposedasasaferalternativetosmoking,leadingtoane-cigaretteacceptabilityincreasebythepublic(CamengaandTindle,2018).Likecigarettes,e-cigaretteshaveturnedintoahugemarketabletosustainmanydifferentproductbrands.Whencomparingtraditionalande-cigarettes,thekeydifferencesistheuseoftobacco.Traditionalcigarettescontaintobaccowhilee-cigarettestypicallydonot.Thisisoneoftheprimaryreasonswhytheyareconsidered“healthier”.Nonetheless,bothproductsstillcontainnicotine.Nicotineistheprimaryaddictiveingredientine-cigarettesolutionsandtobaccoproducts(Walley,2019).Non-scientificclaimsarecreatingconfusioninthepublic’sperceptionofthehealthofe-cigarettes.Theyareinfacthazardoustohumanhealth.Theirusecancausemultiplesymptomsintherespiratory,nervous,gastro-intestinalsystem,etc.(MeoandAsiri,2014).Yet,accordingtotheU.S.FoodandDrugAdministration(FDA),3.6millionyouthstillcontinuetousee-cigarettestoday.PreviousresearchhasdeterminedtheeffectsthatcigarettesmokevaporhasonC.albicans.ThesefindingsarethatcigarettesmokecondensatepromotedC.albicansgrowthandSAP2enzymaticactivity(Semlali,2014).Becausebothmethodsofcigarettesmokingarestillwidelyusedtoday,moreresearchneedstobedonetocontributetopublichealthknowledge.Theobjectiveofthisstudyistocomparetheeffectsoftraditionalande-cigarettesmokevaporonthegrowthrateandSAP2geneexpressionofC.albicans.WehopetorelatethesefindingstotheriskoforalcandidiasiswithotherwisehealthysmokerswhocontainC.albicansintheiroralcavity.Toperformthisexperiment,aninhouse“smokingdevice”(Baboni,2009)willbeusedtocollectcigarettesmokecondensate(CSC)foratraditionalcigarettegrouptreatment.A5%nicotineJuulpodliquidwillbeusedforthee-cigarettegrouptreatment.AGasChromatographyMassSpectrometry(GCMS)analysiswillbeperformedtomeasuretheamountoftracelevelnicotineinbothtreatmentliquidstoensureequalamounts.AlayeroftheCSCandtheJuulpodliquidwillthenbeappliedto10yeastpeptonedextrose(YPD)agarplateseach.Acontrolgroupwithnoadditivetreatmentwillalsobeincluded.C.albicanswillbesuspendedintoaseparateYPDbrothtoensure~200colonyformingunits(CFU)/mL.1mLofthissuspensionwillthenbespreadontothesurfaceoftheplatesandincubatedat37°Ctomaintainitsyeastform.Twoplatesfromeachgroupwillbetestedat12hrs.,24hrs.,36hrs.,48hrs.,and60hrs.afterinoculation.Thistestwillincludecountingthenumberofcoloniesforagrowthassessmentandrunningareversetranscriptasequalitativepolymerasechainreaction(rt-qPCR)assessment.Themeannumberofcolonieswillbecalculatedforeachgroup.Thegrowthrateswillthenbecalculatedusingtheequation:Rate=changeinnumberofcolonies/time.Theseresultswillthenbeplacedintoalinegraphforclarity.Theprocedureforthert-qPCRwillbeimitatedfromasimilarstudyperformedbyHumidahAlanazi,AbdelhabibSemlali,WitoldChmielewski,andMahmoudRouabhiain2019.Firstly,RNAwillbeextractedandconvertedintosingle-strandedcDNAusingapurchasedHigh-CapacitycDNAReversetranscriptionKit.ThereactionswillbepreparedbycombiningthesampleRNAwiththe2XRTmastermix(providedbythekit)andthenplacingthemintothethermocyclerwherethecycleconditionsinclude:10minutesat25°C,followedby120minutesat37°C,5minutesat85°C,andthencoolingto4°C.Next,thesecDNAsampleswillbecombinedwithpurchasedsyntheticnucleotideprimers(Table1:SAP2),andapurchasedSYBRGreenMastermixinordertorunthert-qPCR.Thert-qPCRcycleconditionsinclude:5minutesat95°C,followedby30cyclesof15secondsat95,30secondsat60,and30secondsat72°C.Finally,thecomparativeCtMethodofrt-qPCRwillbeimplementedtoanalyzetheseresults.ThismethodcomparesthedifferencesbetweentheSAP2geneCtvaluesofeachsampleagainstareferencegene(Table1:ACT1)thatexpressesatauniformlevel.Themeanofthesedifferenceswillthenbecalculatedforeachgroupandcomparedusingaone-wayANOVAtest.Table1.PrimerSequencesGenePrimerSequence(3’-5’)Tm(°C)AmpSize(bp)SAP2Forward:TCCTGATGTTAATGTTGATTGTCAAGReverse:TGGATCATATGTCCCCTTTTGT5482ACT1Forward:GACAATTTCTCTTTCAGCACTAGTAGTGAReverse:GCTGGTAGAGACTTGACCAACCA5787

VictoriaSize
ACorrelationStudyofACEs,BCEs,andASD

Epigeneticfactorsplayaroleingeneexpressionintheoffspringofaffectedparents.Atpresent,nosingleenvironmentalconditionorgeneticlinkhasbeenfoundtoproducehumanswithadiagnosisofAutismSpectrumDisorder(ASD).ResultsofthisresearchwillhelpfuturestudyeffortsindeterminingtheexactcauseofASDworldwide.ThisstudywillexamineenvironmentalconditionsofparentsduringtheirchildhoodtodeterminewhetherthereisacorrelationbetweenbenevolentandadverseparentalconditionsandadiagnosisofASDinoffspring.ItisknownthattherearegeneticfactorsthatcanplayaroleinthedevelopmentofASD.ToeliminatetheelementofgeneticheritabilityasthecauseofASD,onequestionnairewillbesuppliedwhichrequestsinformationonsocialpersonalitypreferenceswhichwouldthenscreenparentsforbroadautismphenotype,whichwouldsignalapossiblegeneticinheritanceasthecausefortheirchild’sdevelopmentofAutismAdditionally,participantswillcompletetwomoresurveyswhichasksquestionsaboutnegativeexperiencesduringchildhoodandpositiveexperiencesduringchildhood.ThisdatawillbecodedandanalyzedusingstatisticalsoftwareSPSStodetermineifthereisanepigeneticlinkbetweenparentalchildhoodtreatmentandtheproductionofatypicallydevelopingchildren.Thisepigeneticeffecthasalreadybeenstudiedanddemonstratedusingmousemodelsandfearconditioning.

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Spring2021GrantAwardees

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BrandtEvans
CannabinoidsPotentialasAntibiotics

TheCDClistsantibioticresistanceasthenumberoneglobalhealththreatintheworld1.Certainbacteriasuchasmethicillin-resistantStaphylococcusaureus(MRSA)responsibleforvarioustypesoftissueinfectioninhumans,Staphylococcusaureus(S.aureus)responsibleforskininfectionsandEscherichiacoli(E-coli),abacteriathatcommonlycausesurinarytractinfectionsaregrowingincreasinglyresistanttocurrentantibiotics.TheCDCstatisticsshowthereare2.8millionantibioticresistantinfectionsyearly,resultingin35,000deaths1.Thisiswherecannabiscannabinoidscouldplayavitalroleasnewantibiotics.Cannabinoidsaresecondarymetabolitesfoundinthecannabissativaplantbelievedbymanytohavevariousmedicinalproperties.Thehumanbodycontainsanendocannabinoidsystemthatworkswithcannabinoidstocreatenumeroustypesofsignalsandtriggersintheimmuneandnervoussystemofthebody.Theproposedprojectistotestthreecannabinoids,tetrahydrocannabinol(THC),cannabidiol(CBD)andcannabigerol(CBG)againstMRSA,Staphylococcusaureus,andE-coli.Theprojectwillbeperformedusingminimuminhibitoryconcentration(MIC)andminimumbactericidalconcentration(MBC)assays.TheMICvaluewillbeanumericalrepresentationoftheconcentrationofcannabinoidrequiredtoinhibitthebacteria’sgrowth.TheMBCwillbecalculatedfromtheMICachievedandrepresentstheconcentrationofcannabinoidthatcausesapproximately99.9%bacterialdeath.Theseassayswillcombineaninoculatedbacteriastrainataspecificconcentrationandthecannabinoidatincreasingconcentrationsin96wellplates.ThelowertheMICvaluethebetterandtheclosertheMBCtotheMICthebetterantimicrobialproperties.Theneedtodevelopimprovedantibioticsislongoverdue.Inthisresearchagoalwouldbetoseeallcannabinoidstestedeffectiveagainstbothgram-negative(E-coli.)andgram-positive(MRSA,S.aureus)bacterialstrains.Thiswouldprovideevidencetowarrantfurtherresearchintotheseplantscompoundsasfutureantibiotics.

SimonFreeman
LarvalLakeWhitefish/ZooplanktonDensityatDayandNight

Lakewhitefishsupportsomeofthemostcommercially,culturally,andrecreationallyimportantfisheriesintheGreatLakes.Lakewhitefishpopulationshavedecreasedsincethe1990sintheupperGreatLakes,coincidingwiththeinvasionofzebraandquagga(dreissenid)mussels(Pothovenetal.2001).Theseinvasivemusselshavedecreasedphytoplankton,andsubsequently,zooplanktonpopulations(PothovenandMadenjian2008).Adequateamountsofzooplanktonarecriticalforthegrowthandsurvivaloflarvalfish(Zornetal.2020),includinglakewhitefish,whospendthistimeinshallow,beachhabitats.OnehypothesisfordecreasedlakewhitefishpopulationsintheGreatLakesisaresultofdecreasedzooplanktonpopulations.However,limiteddataexistthatexaminezooplanktonandlarvallakewhitefishrelations,andexistingdataareonlyfromdaytimebeachsurveys.Habitatusebyzooplanktonandfishcanvarysignificantlyatdayversusnight,necessitatingdayandnightcomparisonstodeterminewhetherdaytimesamplingisadequateforquantifyinglakewhitefish-zooplanktonrelations(Doubeketal.2020).Toaddresstheseknowledgegaps,Iask:1)WhatistherelationshipbetweenlarvalLAWandzooplanktonpreydensityinLakesMichigan,Huron,andSuperior?,and2)Doestherelationshipdifferatnight?Toexpandourknowledgeontherelationsbetweenzooplanktonandlarvallakewhitefishpopulations,IwillworkwithLSSUandtheSaultTribeofChippewaIndianstosamplezooplankton,larvallakewhitefish,andenvironmentaldata(watertemperature,pHetc.)at3locationsineachofthefollowing:LakeMichigan,LakeHuron,andLakeSuperior.MysamplingwilloccurbetweenApril-JuneintheEasternUP.Inadditiontothis,Iamreceivingsamplesfrommultipletribal,state,andfederalagenciesthroughouttheGreatLakesregion(GreenBay,Muskegon,Duluthetc.).AllsampleswillbeprocessedbymyselfinthelabatLSSU,andIwillrunstatisticalanalyses.

MackenzieKalchik
TheEffectsofDiaphragmaticBreathingonMelatoninandSleepAmongCollegiateStudent-Athletes

Alongwiththemanyresponsibilitiestheaveragecollegestudentexperiences,collegiatestudent-athleteshaveadditionaltime-consumingresponsibilities.Theseadditionalresponsibilitiesincludepractices,competitions,travel,teammeetings,andmore.Thestressorsofacademicsandathleticshaveproventohavealargeimpactonthewell-beingofstudent-athletes,withalmosthalfofmalestudent-athletesandmorethanhalfoffemalestudent-athletesreportingthatstressfromathleticsandacademicshasimpactedtheirmentaloremotionalhealth(Humphrey,Yow,andBowden,2000).Withthehighprevalenceofstressamongstudent-athletes,itiscommontoleadtoothernegativeimpactsonthebody.Chronicstresscanleadtoadecreaseinquantityandqualityofsleep(Soffer-DudekandShahar,2011).Insufficientsleephasmanyconsequencestothebody,includingtheweakeningoftheimmunesystemandadecreaseinperformanceofthehypothalamus,pituitary,andadrenalglands,whichleadstofurtherconsequencesofadeclineinphysicalperformanceandlearningability(Ghorbaniatal.,2019).Melatoninisthemainhormonesynthesizedbythepinealglandthatiscontrolledbythehypothalamusandplaysaroleinregulatingsleep-wakecyclesbyreleasinglowlevelsduringtheday,andthehighestlevelsatnighttopreparethebodyforsleep(Jennumetal.,2016).Becausestresscanimpacttheperformanceofthehypothalamus,thereforeimpactingthesecretionofmelatonin,circadianrhythmcanbedisrupted,consequentlyinterferingwithsufficientsleepinthoseaffected.Duetothedamagesthatstressandlackofsleepcanhaveonanathlete’sperformanceintheclassroomandincompetition,itisimperativetogiveathletesatechniquetheycanutilizeonandoffthefieldofplay.Diaphragmaticbreathing(DB)isatechniqueusedbymanyyogaandmeditationpracticesthatinvolvesbreathingdeeplyintothelungsbyexpandingthediaphragminsteadoftheribcage(Martarellietal.,2009).Thisbreathingtechniqueutilizestechniquesthatstrengthenthediaphragm,decreasetheworkofbreathingbyslowingrespirationrate,anddecreaseoxygendemand(ClevelandClinic,2018).Diaphragmaticbreathingexerciseshaveproventobeanon-invasivetreatmentthatpositivelyimpactsinsomnia,cardiacautonomicfunction,depression,andanxiety,allofwhichcouldimprovethementalandphysicalhealthofanathlete(Ghorbanietal.,2019).Notonlydoesdiaphragmaticbreathingimprovetheseimportantaspectsofathletehealth,butitalsohasbeenproventoreducetestingstressandincreasetestingperformanceofthegeneralcollegestudentpopulation(Paul,Elam,andVerhulst,2007).Thepurposeofthisexperimentistoinvestigatetheimpactofdiaphragmaticbreathingexercisesonsleepinthecollegiatestudent-athletepopulationbytestingandobservingmelatoninlevels.Participantswillbegivenafive-minutediaphragmaticbreathingexerciseandaskedtoperformiteverydayforaperiodoftwoweeks.Melatoninlevelswillbecollectedoncebeforetheybeginthediaphragmaticbreathingexercises,andagainafterthetwoweeksofcompletingtheexercises.Itishypothesizedthatsleepquantityandqualitywillimproveamongthestudent-athletesperformingthebreathingexercisesandwillbereflectedintheincreaseofmelatoninlevels.Methods:Agroupoftwenty-fourstudent-athletesatLakeSuperiorStateUniversitywillbeaskedtovolunteerinthisstudy.Theywillbegiveninstructionsonthediaphragmaticbreathingexercisestheywillbeaskedtoperform,andwrittenconsentwillbeobtained.Therewillbeacontrolgroupoftwelvestudents-athletesthatwillnotperformdiaphragmaticbreathingexercises,andanexperimentalgroupoftwelvestudent-athletesthatwillperformthebreathingexercises.Datawillbecollectedintwoways,whichwillconsistofasurveyandmeasurementsofmelatoninlevelsthatwillbecompletedbeforeandafterthestudy.Eachparticipantwillbeassignedanumericalcodetomaintainconfidentialityoftheparticipants.Theexperimentalgroupwillbeshownafive-minutediaphragmaticbreathingexercisetheycanfindinanymobileAppStorecalled,“StrategicBreathing,”thatwascreatedbyMaxStrom,anauthorandteacherwithmanyaccoladesregardinghisworkwithdiaphragmaticbreathing(Strom,2021).Theexercisewithintheappincludesvisualandaudioinstructions.Theexperimentalgroupwillbeaskedtoperformthebreathingexercise30minutesbeforebedeverynightforthetwo-weekperiodintheirownresidency(Chander,2020).Thecontrolgroupwillnotbeaskedtoperformthebreathingexercisesandtocontinueonwiththeirdailyhabits.Thesurveythatwillbeconductedwillaskaseriesofquestionsthatwillbebeneficialtodeterminetheeffectivenessofthebreathingexercisesfromasubjectivepointofviewwithintheparticipants.AdaptedfromAssessmentofRecoveryafterScheduledTimeofftoHospitalEmployeesWorkingNightShift,questionswillbeaskedaboutsleepquality,sleepquantity,athleticperformance,academicperformance,andlevelsofstressparticipantsexperience.(Blyly&RansonOlson,2018).Thesesurveyswillbegivenbeforeandafterthestudytoeachparticipantinboththecontrolandexperimentalgroup.Tomeasuremelatoninlevels,amelatoninsalivaELISAAssaykitwillbeused.Salivawillbecollectedbyunstimulatedpassivedrool.Participantswillbeaskedtonoteat60minutesbeforesamplecollection,aswellastoavoidconsumingalcohol12hourspriortosamplecollection.Toanalyzethedata,at-testwillbeusedtoobservethechangesinmelatoninlevelswithineachindividualfromthebeginningofthetwo-weekexperimentcomparedtotheend.Anothert-testwillbeconductedtotesttheaverageeffectbetweentheexperimentalandcontrolgroups.AnANOVAwillbeperformedtodetermineanysignificantdifferences.Ifthereisasignificantdifferencethatsupportsthehypothesisthatdiaphragmaticbreathingincreasesmelatoninlevels,thereforeimprovingsleep,theresultswillbepresentedtocoachingstaffsthroughouttheathleticdepartmentatLakeSuperiorStateUniversity.Ifimplemented,diaphragmaticbreathingcouldbeabeneficialtoolforstudent-athletestoimprovetheirphysicalandmentalhealth.LiteratureCitedBlyly,McKenna,B.RansonOlson.2018.AssessmentofRecoveryafterScheduledTimeofftoHospitalEmployeesWorkingNightShift.Chander,T.,LakeviewInternalMedicine,personalcommunication.ClevelandClinic.2021.Diaphragmaticbreathingexercises&techniques.

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Fall2020GrantAwardees

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BobbiJoCaskey
ImpactofPlantSpeciesontheMicrobialCommunityofanAquaponicSystem

Aquaculture,Hydroponics,andAquaponicsAquacultureisthetermusedtodescribethebreeding,raising,andharvestingoflivingorganismsinanaquaticenvironment(USDepartmentofCommerce,2019).“Fishfarming”isoftenacommonterm.Acommonexampleofthisisrecirculatingaquaculturesystem(RAS).Whilesystemsmayvary,theconceptremainsthesame.Thewaterthatholdsthefishcirculatesoutalongwithanywasteorleftoverfood.Itisthenbroughtthroughafiltration,thenanaerationunitthatwillreturnoxygentothewaterandremovecarbondioxide(Losordo,et.al.,1999).Fisharenottheonlyorganismthatcanberaisedasalgae,seaweed,andotheraquaticplantscanalsobegrown.Hydroponicsisasimilarconcept,howeverterrestrialplantsareproduced.Itincludesgrowingcropsinwaterratherthansoil(Tripp,2014).Fertilizers,includingvitaminandmineralsareaddedtothewatertoensuretheplantshavewhatisneededforgrowth.Whilebothhydroponicsandaquaponicsprovidesignificantadvantages,specificallywhenaddressingoverconsumptionofourfoodsupply,recentstudieshaveshownthatacombinationofthetwocouldprovideanevengreaterbenefit(Jungeetal.,2017).Aquaponics,nottobeconfusedwithaquaculture,integratestherecirculationofwaterandaerationofaRASbutusestheterrestrialplantsandnitrifyingbacteriafromhydroponicstoridthesystemofwasteproductsharmfultothefish.Thefishwillproducenitrogenwasteintheformofammonia.Thisammoniaisthenbrokendownintonitriteandthennitratebythemicrobes.Abuildupofammoniaornitritecanbeharmfultothefishhowevernitrateisrelativelyharmless.Nitrateisalsothepreferredsourceofnitrogenforplants(Rakocy,etal.,n.d).Intheendtheplant,fish,andbacteriaallbenefit.MicrobesintheSystemWhileallcomponentsofthesystemareimportant,thisstudywilllookspecificallyatthemicroorganisms.Hydroponics,likeaquaponics,oftenusenitrifyingbacteriatoproducenitrate,soitcanbeassumedthatthemicrobiotawillbesimilar,howeveritisunclearonhowtheaquacultureaspectwillaffectthesemicrobes.Itisnotunlikelytofindplantgrowth-promotingrhizobacteria(PGPR)inahydroponicsystem.Rhizobacteriaarethebacteriathatgrowdirectlyontheedgesoftheroots.Thismeansthattheyhavedirectcontactwiththeroots.Theymaintainasymbioticrelationshipandhelppromoteplantgrowthaccomplishingoneofthefollowing:reducingnitrogenintonitrate,enhancingstressresistance,stabilizingphosphateandpotassiumintake(Cakmakci,Donmez,andErdoan,2007).Whiletheyareextremelycommoninsoil,PGPRsarealsousedasabiofertilizertohelpplantgrowthinbothhydroponicsandsoilgrowncrops.ItwasspecificallyfoundthatBacillusspp.Gliocladiumspp.,Trichodermaspp.,andPseudomonasspp.werecommonbacteriaandcanbefoundinahealthyhydroponicsystem(Lee,andLee,2015).AllthepreviousspeciesarealsoconsideredRhizobacteria.Eachspeciesbenefitstheplantdifferently.Forexample,onedistinctionofBacillussppisthatsomespeciesloweredtheinfectionrateofsomepathogens(Nihorimbereetal.,2011).Inaddition,specificspecieswerereportedtoincreasewaterqualityandgrowthrate(Nihorimbereetal.,2011).SomespeciesofPseudomonasspp.wereshowntodecreaserootrotandhavesomeantifungalpropertiesoncertainspecies(McCullagh,etal.,1996).Nowturningtheattentiontotheaquacultureside,onestudyfoundthattwomaintypesofbacteriawerefound:autotrophicandheterotrophicbacteria.ManyautotrophicbacteriawereammoniaoxidizerssuchasNitrosococcus,Nitrosospira,andNitrosomonasornitrateoxidizerssuchasNitrobacterandNitrospira(RurangwaandVerdegem,2015).Theseoxidizershelpreducetheamountofammoniaandnitrite(thatareharmfultothefish)intonitrateornitrogengasthataremuchlesstoxic.Theheterotrophicbacteriawerefoundtobedifferentdependingonthespeciesoffish.ForexampleonestudyusingcarpfoundAlphaproteobacteriaandBetaproteobacteria.LaterthesamegroupusedgoldfishinsteadandfoundalargervarietyofmicrobesthatincludedActinobacteria,Bacilli,Gammaproteobacteria,Planctomycetacia,Sphingobacteria,Hyphomicrobiumdenitrificans,RhodovulumeuryhalinumandNitrospiramoscoviensis(Sugita,etal.,2005).AdvantagesofAquaponics/ImplicationsandRelevanceFoodsecurityisbecominganincreasinglyrelevanttopicglobally.TheUnitedNationsFoodandAgricultureOrganization(FAO)estimatedthatby2050,theglobalpopulationwillreach9.1billionandouragriculturalsystemallonewillnotbeabletosupplyenoughfood(Sohngen,2017).Inaddition,studieshaveshownagreatdeclineinlaborinagriculture(Christiaensen,Rutledge,andTaylor,2020).Finally,astheamountofpeoplegoingintoagriculturedecreases,itwasalsofoundthatthosegoingintotheagriculturalsectorhavelessknowledgethanpreviousgenerations(Kuiper,Shutes,VanMejil,etal.,2019).Withouttheproperknowledgeandunderstanding,aseriesoftrialanderrorswilloccurthatcouldhavebeenprevented.Nowthatweareawareoftheproblem,itisnowtimetocomeupwithasolution.Aquaponics,aspreviouslymentioned,recirculationofwateralongwiththeaerationofaRASbutusestheterrestrialplantsandnitrifyingbacteriafromhydroponicstoridthesystemwasteproductsharmfultothefish.Theendresultwouldbebothasupplyoffishandproducegrownfromtheplants.Aquaponicsalsohasamultitudeofotherbenefits.Someagriculturalpoliciessuchaspesticides,fertilizers,andoveruseofsoilhaveshownnegativeeffectsontheenvironment(GoudieandViles,2003).Aquaponicsrecirculatesthewaterandonlyproducesorganicwastethatiseasilybrokendown(Konig,etal.,2016).Thismakesaquaponicsaneco-friendlysolution.Aswellasbeinganenvironmentallyfriendlysolution,itisalsoeconomicallyfriendly.Thelargestcostofmaintaininganaquaponicsysteminacommercialstylesystemwasfishfood(Konig,etal.,2016).Theaerationunit,bacteria,andplantscontributetomakethewaterreusableforthefishmeaningthecostofwatersignificantlydecreases.Finally,aquaponicbringsforththeideaofhavingalocalfoodsourceratherthanhavingdependenceonfoodbeingbroughtin.Thisdependencecanbeseencurrentlyinthepandemic.Studieshaveshownthatbecauseoftravelrestrictions,someconsumersarenolongerabletovisittheirtypicalspotnorweretheirtypicalproductsavailable(Laguna,et.al,2020).Itisalsopredictedtoseean17millionincreaseinfoodinsecureAmericansin2020comparedto2018(Gundersen,etal.,2020).Ifusedaccordinglyaquaponicscouldhelpreducethedependenceontransportedgoods.Aquaponicscouldbeonepartofsolvingtheproblemoffoodinsecurity.Ithasshowntohavegreatpromiseinmultipleaspectseconomically,environmentally,andevenmoresoinfoodsustainability.However,itsfullpotentialcannotbereachedwithoutathoroughunderstandingofthesystem.Abundantresearchhasbeendoneonthemicrobialecosystemofhydroponicsandaquaculture,butlittlehasbeendoneinaquaponics.Traditionalaquaculturedoesnottypicallylookintothemicrobesofthesystem.OneofthemainreasonsbeingthatDNAsequencingisveryexpensivetobeproforme.In2015,itwasestimatedthatawholehumangenomesequencewouldcostapproximately$4,000(Wetterstrand,2020).Thisstudywillcontributetotheknowledgeonthemicrobialecosystem.Theobjectiveofthisstudyistoidentifycommonmicrobesofanaquaponicsystemandtoseeifthetypeofplanthasanyeffectonthemicrobeswithinthesystems.Thehypothesisisthatwhiletherearesomemicrobesthatarepredictedtobethesame,therewillbeasignificantdifferencebetweenthesystemswithdifferenttypesofplants.Methods:OverviewofsetupSeven(7)five-gallontankswillbeusedinthisstudy.3tankswillhaveplantA(mostlikelybasil),and3tankswillhaveplantB(mostlikelyswisschard).Theremaining2tankswillbeacontrolandholdadditionalfishrespectively.Thesefishwillbeusedasareplacementifoneofthefishinthetrialsystempasseson.Alltankswillbecleanedwith20%bleachandthoroughlywashedoutwithdeionizedwater.Theywillthenbeairdriedfor7daystoensurethatanyleftoverchlorineisdegraded.Thefigurebelowdepictsalargerversionofthesystemthatwillbeimplementedinthisstudy.Itwilluseacylindricaltubingcuttolooklikea“C”toholdtheplantsgrowninrockwool.Asubmersiblewaterpumpwillbeusedtoforcewaterintothetubing.Thetubingwillbeheldatanangletoallowwatertoflowbackintothetank.Tospeedupthegrowthofbacteria,nitrifyingbacteriawillbeaddedasanadditive(thisiscommoninaquaculture).Threegoldfishwillbeplacedineachtrialtankalongwiththecontrol.Thefishwillbefed1.2gramsoffood.Measurementsoftemperature,ammonia,nitrite,nitrite,pH,chlorine,waterhardness,anddissolvedoxygenwillbetakeneveryMonday,Wednesday,andFridaybeforetheadditionoffoodtothesystem.Thiswillbedonetorecordandunderstandthehealthofthesystemwhilethemicrobiotaisbeingestablished.Thissystemwillbeallowedtorunfor45-60days.Aftertheallottedtime,microbialsampleswillbecollected,andanalyzed.ThissystemwasdesignedbyagroupofNewJerseystudentsfortheLunarPlantGrowthChallengeputonbyNASA.ImageCredit:AtlanticCountyInstituteofTechnology(Smith,2009)MicrobialAnalysisInordertoidentifythemicrobes,thefollowingprocedurewillbeperformed.Asampleofwaterfromeachtankwillbecollectedinasterilizedbeaker.Duringtransportationacleantinfoilcoverwillbeusedtopreventcontamination.Thesampleswillthenbeimmediatelyfilteredusingafiltermembranewith.45micrometerpores.Thisislargeenoughtocatchanyorganism,yetstillbeabletofilteroutanywaste.TheDNAwillthenbeextractedfromthefilterbyusinganIllustraBacteriaGenomicPrepMiniSpinKit.Onceextracted,PCRwillthenbeusedalongwitha16smicrobiomekitthatwillhelpenrichandamplifythebacterialDNA.Afterthis,DNAClean&Concentrator™Kits,ZymoResearchwillthenbeusedtopurifythebacteriaandensurethatanyleftovermaterialfromthePCR.Finally,DNAsequencingwillbeabletobeperformedandthedatawillthenbeanalyzedtoidentifythespeciesofmicrobesDataAnalysisOncethemicrobesareidentified,aseriesoftwogroupt-testswillbeperformed.Sincethedatawillnotbenumbers,somemodificationswillhavetobemade.Tobegin,eachsystemwithplantAwillbepairedwithasystemformwithplantB.Inalltherewillbe3pairs.Then,allthefoundmicrobeswillbeplacedonaspreadsheet.Atthispoint,themicrobeswillbetranslatedinto1or0dependingifthemicrobewasfoundinthesystem(1),ornotfoundinthesystem(0).EachpairwillthenhaveatwogroupT-testrunonthemtoseeifthereisanysignificantdifferencebetweenthetwo.Afterthecalculationsarecompleted,confidenceintervalswillbeusedandaconclusionwillbemade.Timeline:Fall2020:(current)prepareproposal,research,trialsystems,presentproposalSpring2021:continuetrialresearch(determiningifthisisthesetupIwanttous),practicejjjjjjjjjjjjjjjjjjjmaintainingsystem(toensurelessproblemduringexperimentation)Summer2021:WorkandearnmoneyforprojectFall2021:Performproject,gatherandorganizedata,beginanalyzingresultsSpring2022:takeBio499,beginfinalreport,seniorpresentationBudget:Fish:.25$each$26inoneteststrips:100for15$$15Submersiblewaterpumpswithtubing$845-gallontank8x10$80DNAClean&Concentrator™Kits,ZymoResearchpresentinlab16sMicrobialDNAprepkitpresentinlabPCRreagents$~50ZymoBiomicsDNAminiprepkit$274ExtractionfilterpresentinlabVariousglasswarepresentinlabPCRmachinepresentinlabPlasticpetridishes(40)$22NutrientAgarpowder$46.30Flowcell-NGSNanopore$100Overallprice:$673.30Anysuggestionswouldbeextremelybeneficial,andIthankyouforyourtime.LiteratureCited:Cakmakci,R.Donmez,M.andErdoan,Ü.2007.TheEffectofPlantGrowthPromotingRhizobacteriaonBarleySeedlingGrowth,NutrientUptake,SomeSoilProperties,andBacterialCounts.TurkishJournalofAgricultureandForestry31:189-199.Christiaensen,L.Rutledge,Z.andTaylor,J.E.2020)Viewpoint:Thefutureofworkinagri-food.FoodPolicy,101963.Goudie,A.andViles,H.2003.Theearthtransformed:Anintroductiontohumanimpactsontheenvironment.JohnWiley&Sons,NewJersey,USAGundersen,C.Hake,M.Dewey,A.etal.2020.FoodInsecurityduringCOVID‐19.AppliedEconomicPerspectivesandPolicy00,00:1-9Junge,R.König,B.Villarroel,M.Komives,T.etal.2017.StrategicPointsinAquaponics.Water.MDPI.9(3):10.3390/w9030182.Konig,B.Junge,R.Bittsanszky,A.etal.2016.Onthesustainabilityofaquaponics.Ecocycles2(1),26-32.Kuiper,M.Shutes,L.VanMejil,H.etal.2019.Laborsupplyassumptions-Amissinglinkinfoodsecurityprojections.GlobalFoodSecurity25.Laguna,L.Fiszman,S.Puerta,P.etal.2020.TheimpactofCOVID-19lockdownonfoodpriorities.ResultsfromapreliminarystudyusingsocialmediaandanonlinesurveywithSpanishconsumers.FoodQualityandPreference86,104028.Lee,S.andLee,J.2015.Beneficialbacteriaandfungiinhydroponicsystems:Typesandcharacteristicsofhydroponicfoodproductionmethods.ScientiaHorticulture195:206-215.Losordo,T.Masser,M.andRakocy,J.1999.RecirculatingAquacultureTankProductionSystemsAReviewofComponentOptions.SRACPublicationNo.453.McCullagh,M.Utkhede,R.Menzies,J.G.etal.1996.Evaluationofplantgrowth-promotingrhizobacteriaforbiologicalcontrolofpythiumrootrotofcucumbersgrowninrockwoolandeffectsonyield.EurJPlantPathol102:747–755.Nihorimbere,V.Cawoy,H.Seyer,A.etal.2011.ImpactofrhizospherefactorsoncycliclipopeptidesignaturefromtheplantbeneficialstrainBacillusamyloliquefaciensS499.FEMSMicrobiologyEcology79(1):176-191.Rakocy,J.Masser,M.andLosordo,T.(n.d).RecirculatingAquacultureTankProductionSystems:Aquaponics—IntegratingFishandPlantCulture.SRAC-454.Rurangwa,E.andVerdegem,M.C.2015.Microorganismsinrecirculatingaquaculturesystemsandtheirmanagement.ReviewsinAquaculture7(2):117-130.Smith,H.2009.Aquaponics.RetrievedNovember01,2020fromSohngen,T.2017.TheWorldMayRunOutofFoodintheNextDecade:Study.RetrievedOctober15,2020.Sugita,H.Nakamura,H.andShimada,T.2005.Microbialcommunitiesassociatedwithfiltermaterialsinrecirculatingaquaculturesystemsoffreshwaterfish.Aquaculture243(1-4):403-409.Tripp,T.2014.Hydroponicsadvantagesanddisadvantages.ProsandConsofHavingaHydroponicGarden.SpeedyPublishingLLC,Delaware,USA.USDepartmentofCommerce.2019.Whatisaquaculture?RetrievedOctober06,2020fromWetterstrand,K.2020.TheCostofSequencingaHumanGenome.RetrievedNovember10,2020.

RachelFarina
ImmunoglobulinSpecificityTestingofSalmonidSpeciesUsingRabbit,Rat,andChickenAntiserum

Immunoglobulinspecificitytestingofsalmonidspeciesusingrabbit,rat,andchickenantiserumRachelFarinaLakeSuperiorStateUniversitySchoolofScienceandMedicineDr.JunLiFall2020IntroductionIntheGreatLakesregion,severalspeciesofsalmonidfisharenaturallycommoninthewildandinhatcheriesforresearch,breeding,andhumanconsumption:laketrout(Salvelinusnamaycush),rainbowtrout(Oncorhynchusmykiss),browntrout(Salmotrutta),andatlanticsalmon(Salmosalar).Immunologicalfishhealthisaveryimportanttopicdiscussed,especiallyinplaceswherefishingisamainindustry.Sincethesespeciesoffishliveinsuchlargenumberedcommunities,pathogenscanspreadratherquickly.Largeamountsofdisease,illness,anddeathinfishcaneconomicallyharmthefishingindustrygreatlyandreduceacommonsourceoffoodforthehumanpopulationlivinginthisarea(Magnadottir,2010).Asthefishindustryincreases,theimmunesystemofthesesalmonidspeciesmustbestudiedfully,andlaboratoryreagentsandpracticesmustbedevelopedfurther.Mostorganismshaveanimmunesystem,andfisharethefirstgroupoforganismswithbothinnateandadaptiveimmunity(Magnadottir,2010).Theimmunesystemofanyorganismiscomposedofmanydifferenttypesofcellsandproteinsthatprotecttheorganismfrompathogenicthreats.Thesalmonidimmunesystemdependsontheinnateimmunesystemheavilyforhealth,muchmorethanadaptiveimmunity(Uribe,2011).Complement,aseriesofsolubleproteins,andneutrophils,cellswithphagocyticcapabilities,aresomeofthemaininnatedefensesthatarepresentinfishmucosalmembranestofightoffandkillpathogenicorganisms.Theinnatesystemdoesnothaveanylong-termmemoryfunctions,butitcanfightoffanypathogensveryquicklywithoutneedingspecificantigeninteractions(Ma,2020).Usually,anorganismgetsinfectedwithapathogenandisneverawarebecausetheinnateimmunesystemkillsthepathogenbeforeitcanbecomesymptomatic,andthustheadaptiveimmunesystemisneveractivated.Salmonidsdoalsohaveanadaptiveimmunologicalresponsethatislonglasting,butslowtoactivate.FishhaveB-lymphocytesandT-lymphocytesboth,butnolymphnodesforallthecellstocongregateandacceleratetheproductionofantibodies(Magnadottir,2010).AsingleantibodyisaY-shapedmoleculewithvariableregionsthatallowsdifferentmoleculestointeractwithaveryspecificantigen.B-cellsinfishsecreteantibodiesinresponsetospecificantigensfrompotentialpathogens.Then,theantibodiesfindtheantigensinthebodyandattachtothemsootherimmunecellsknowwhattoattack(Owenetal.,2009).Itisimportanttonote,especiallyintheGreatLakesarea,thatthesecretionofantibodiesistemperature-dependentandtheactivityisloweredincoldtemperatures(Dixon).IgMantibodiesarethemostpresentisotypeinsalmonids,buttheyaretetrameters,fourY-shapedunitsattachedtogether,insteadofpentameters,fiveY-shapedunitsattachedtogether.ThepentameterIgMisfoundinhumansandmammals(Magadan,2016).Theadaptiveimmunesystemneedstobeactivatedbytheinnatesystem,soanorganismwouldonlyproduceantibodiestosomethingthattheinnateimmunesystemcouldnothandlebyitself.Theadaptiveimmunesystemcontainsamemorysystemwhichpreventsthefishfromgettingsickfromthesamepathogentwiceandafasterimmuneresponsetofamiliarpathogens(Owenetal.,2009).Becauseofthis,evenhealthyfishcontainantibodiesintheirserum.EnzymeLinkedImmunoassayAssay(ELISA)testingisamajortechniqueinimmunologicallaboratories.Ittestsforthepresenceofaspecificantigenorantibodybyusingtheoppositeasareagent.Antibodiesareveryspecificandmatchwithonlyoneantigenwithgreatspecificity.Theoriginalserumandaknownlaboratoryreagentaremixedtogetherintoawellandallowedtoincubatesotheantigenandtheantibodiescanbindtogether.Insomecases,asecondarydetectionantibodyisnecessary,whichis“a[n]antibodythatbindsaprimaryantibodythatisnotenzyme-conjugated”(Alhajj,2020).ThisistheindirectELISAtest.Then,acolorchangingsubstrateisadded,usuallyhorseradishperoxidase(HRP),sothewellscanbeanalyzedinaplatereader(Owenetal.,2009).Theconcentrationofantibodiesdetectedcanbedeterminedfromacreatedstandardcurveusingtheabsorbancereadings.Antiglobulintestingisaprocedureusedinmicrobiological,virological,andimmunologicallaboratories.Itisusedtodetectthepresenceofantibodiesinonespeciesbyusingtheantiserumofanotherspecies.Purifiedantibodiesfromtheoriginalserumthatisbeinganalyzedareinjectedintoasecondaryspecies,wheretheanimal’simmunesystemcreatesantibodiesagainstthepurifiedantibodiesthatwereinjected(Harmening,2019).TheseantibodiescanbeusedasthesecondarydetectionantibodiesinanindirectELISAtest(Alhajj,2020).Thiscanbeusedtodetectthepresenceofaspecificantibodyinalaboratorysettingandtheminimumdilutionthatdetectstheantibodies.Inthisspecificstudy,IgMantibodieswerepurifiedfromthesalmonidspecies,andanti-fishIgMIgGantibodieswerecreatedfortherabbitandrat,andanti-fishIgMIgYantibodieswerecreatedforthechicken.Thisstudywilltestthespecificityofeachantibody-antiglobulinreactionbetweenthefoursalmonidspeciesandthethreedifferentantiglobulinproducingspecies.Serumalreadycollectedfromeachfishspecieswillbeusedforallspecimens,andalloftheantiserumisalreadycreatedandisstored.ThesesamplesarefrompreviousstaffandstudentsofLakeSuperiorStateUniversity.ThestudywilldeterminewhichantiseraworksbesttodetectIgMantibodiesforeachsalmonidspeciesandtheminimumdilutionatwhichtheyaredetected.MethodsandAnalysisThedetectionmethodtodeterminethespecificityofeachcombinationofspecimenandantiserumwillbedonebyELISAtesting.ThesalmonidIgMantibodieswillbeintroducedintoawellplate,andincubatedfor15minutesatroomtemperaturetoallowthemtocoatthewallsofeachwell.Then,thedifferentantiserasecondaryantibodieswillbeintroducedtothewells,andallowedtoincubatefor15minutesat37degreesCelsius.Last,substratewillbeaddedtoeachwell,incubatedatroomtemperaturefor5minutes,andobservedforcolorchangeresults.Inbetweeneachstep,thewellswillbewashedwith1xPBSthreetimestoensureaccurateresults.Theplateswillbespectrophotometricallyanalyzedbyaplatereaderata450nmtotestforspecificity(Alhajj,2020).Iwilluse96-wellplatesforanalysis.RowAandBwillcontaincontrolmaterials,withfetalbovineserum(FBS)asanegativecontrolandRainbowTroutserumasapositivecontrol.Theothercolumnswillbereplicatesofeachfishserum-antiseracombination.Eachhasaseriesofdilutions,repeatedfourtimes:1:500,1:1000,1:2000,1:4000,1:8000,1:16000.ThissetupcanbeseenbelowinFigure1.Thegoalofthis,isnotonlytoseewhichantiseracandetectthefishantibodies,butalsotoseetheminimumconcentrationneededfordetection.Creatinglaboratoryantiseraandisolatingtheantibodiesforclinicalusearebothtimeconsumingandexpensivetechniques.Itisveryresourcefultoknowwhattheminimumconcentrationneededis,solabreagentscanlastlongerandbeusedmore.IplantouseanANOVAtesttodetermineifthereareanysignificantdifferencestoproveoneantiseraspeciesbindsmorespecificallytothefishantibodiesthantheothers.Sincetherearetwoindependentvariables,atwo-wayANOVAtestwillhavetobeperformed.Thespecificitywillbedeterminedforallinteractions,withtwogroups,orfactors:fishserumspeciesandantiserumspecies.Therearefourlevelsforthefishserum,andthreelevelsfortheantiserumspecies.Thismeansthatdegreesoffreedomis3forthefishserumand2fortheantiserumspecies.Table1:TreatmentGroupsLakeTroutRainbowTroutBrownTroutAtlanticSalmonRabbitRatChickenEachwillhavetheexactsameamountofreplicates,four,andtheaveragesdeterminedbytheplatereaderwillfillinthetablelistedabove.Amaineffecttest,interactiontest,withinvariationtest,andanF-testwillberunusingatwo-wayANOVAtesttodeterminesignificanceofthisstudy.Iftherearesignificantresults,thisprojectmaydevelopfarther.ReferencesAlhajj,M.,Farhana,A.(2020).EnzymeLinkedImmunosorbentAssay(ELISA).Statpearls.Dixon,B.Theimmuneresponseinfish:Abriefreview.Receivedon04October2020fromhttps://www.vin.com/apputil/content/defaultadv1.aspx?pId=11257&id=3863675&print=1Harmening,D.M.(2019).ModernBloodBanking&TransfusionPractices.F.A.Davis,p.103-110.Ma,S.(2020).EffectsofenvironmentalstressorsontheinnatefunctionofAtlanticSalmon(Salmosalar).LakeSuperiorStateSeniorProject.Magadan,S.,Sunyer,O.J.,Boudinot,P.(2016).Uniquefeaturesoffishimmunerepertoires:particularitiesofadaptiveimmunitywithinthelargestgroupofvertebrates.ResultsProblCellDiffer.57:235–264.Magnadottir,B.(2010).Immunologicalcontroloffishdiseases.Marinebiotechnology,12,p.361–379.Owen,J.,Punt,J.,Stranford,S.(2009).Immunology.TheJournalofExperimentalMedicine,p.660-663.Uribe,C.,Folch,H.,Enriquez,R.,Moran,G.(2011).Innateandadaptiveimmunityinteleostfish:areview.VeterinarniMedicina,56,(10):486–503.

VeronicaShull
TeamSuperiorAcousticsandGeophysicalSolutions:AMethodofOilDetectioninBodiesofWaterCoveredbyIce

TeamSuperiorAcousticsandGeophysicalSolutionsisaseniorresearchgroupconsistingofthreeMechanicalEngineeringstudents:________;andoneElectricalEngineeringstudent:____.ThestudentsinthisgrouphavetakencourseworkinAcoustics,SignalProcessing,VibrationsandNoiseControl,ResearchMethodsandElectromagneticstopreparefortheresearchtobepursuedinthisproject.Theteamintendstoperformexperimentstotestamethodofoildetectioninbodiesofwatercoveredbyicewiththepurposeofdetectingandmappingoilspillstoassistinexpeditingthecleanupprocess.ThereislittleresearchavailableinthisareaandduetothecloseproximityofLakeSuperiorStateUniversitytoEnbridge’sLine5,a4.5milelongpipelinecarryingoilthroughtheStraitsofMackinac,thisissueisofgreaterinteresttostudentsattheuniversityandthewidercommunity.Theproposedmethodinvolveshydroacousticsandusesreverberationtimetodetectchangesfromapristinestate.Thehypothesisofthemethodstatesthattherate,atwhichasoundfielddecayschanges,basedonenvironmentalfactors,andtheeffectsofoilonthesoundfieldaremeasurable.Testingofthemethodbythisteamwillbedoneinascaledtankenvironmentandeffortswillbemadetoimplementthemethodinafull-scaleenvironment.Theteamwillalsoimprovethescaledtankenvironmentbyobtainingamaterialthatwouldadequatelyrepresenttheacousticpropertiesoficewithoutrequiringthegenerationofnaturalice,permittingtestingtocontinuewhentheoutdoortemperatureisn’tlowenoughforgrowingicebyexposure.Animpulsivesoundsourcewillalsobeidentifiedbythisteamtobeusedinboththescaledtankenvironmentandinfull-scaleimplementation.Inordertomitigateissuesencounteredbyfuturegroupswithfullscaleimplementation,thisteamwillalsocollectdatafromlakeenvironmentsinordertoanalyzetheambientnoisepresenttoestablishasoundfloorandinvestigatepotentialpassivesources.Inlieuofusingcrudeoilinalakeenvironmentfortesting,anacousticssoftwarepackagewillbeobtainedinordertoperformcalculationsthatwillfurthervalidatethemethod.Documentationofdecisionsmade,supportingresearchandprocessingcodewillbecompiledforfuturestudentstocontinuevalidationandimplementationofthemethod.Thisdocumentationwillbepresentedatanacademicconferenceandapaperwillbepublishedtocontributetothelargerbodyofresearchinoildetectionmethods.Theteamhasabudgetof$1000fromtheSchoolofEngineeringandTechnologyatLakeSuperiorStateUniversitywhichwillbeusedtoobtainmaterialstotestasanicesubstitute,researchpapers,makerepairstothesmall-scaledtankenvironmentandmakeimprovementsonthecurrentsoundsourceusedinthesmall-scaletankenvironment.

CarlieWeaver
TheEffectsofPFASontheBeneficialGutBacteriaLactobacillusAcidophilus

Perfluoroalkylsubstance(PFAS)contaminationhasrecentlybecomealargetopicofdiscussion(andaction)acrossthenation,specificallyinthestateofMichigan.Thesechemicalshaveincitedexecutiveorders,actionresponseteams,andstate-declaredstatesofemergency(MPART,2019).PFASarelong,fluorinatedcarbonchainsthatwerecommonlyusedinindustryfromthe1950suntildiscontinuationintheUnitedStatesaround2015asfoodpackaging,nonstickcoating,firefightingfoam,andanimportantcomponentofteflon.Duringtheseyears,thesechemicalshavefoundtheirwayintogroundwatersources,andfromthereintothehumanbody(Bucketal.,2011).WatersourceshavebeentestedandhavebeenfoundtohavemuchhigherconcentrationsofPFAS,particularlyperfluorooctanoicacid(PFOA)andperfluorooctanesulfonicacid(PFOS),thanisconsideredsafe.Measurementsofgreaterthan30,000partspertrillion(ppt)havebeenfoundingroundwaterdespitethelegalcapforsafetybeingplacedatamere70ppt(Readeetal.,2019).Recently,theCenterforDiseaseControlhasnamedPFAS“chemicalsofconcern”towardhumanhealth,asresearchhasshownthattheymaybeoneofthecausesofnumerousdiseases,includingcancer,diabetes,infertility,andweakenedimmunesystem.Theymayalsocontributetochangesintheliver,thyroid,pancreas,andmore,asthechemicalshavealonghalf-lifeinthebodyandarepresentinserum.(Anderson-Mahoneyetal.,2008)(CenterforDiseaseControl,2018).WhilePFOAandPFOShavebeendiscontinuedintheUS,anewchemicalhastakenitsplace.Named“GenX,”2,3,3,3-tetrafluoro-2-(heptafluoropropoxy)propanoicacidisnowbeingmanufacturedtocompensateforthelossofPFOAandPFOS.Thischemicalhasbeenadvertisedassafe,despitecausingsymptomsofdiseaseinlabrats(Shea,2018).Thecreatorcompany,Chemours,hasendorsedandproducedthechemical.RecenttestsshowthatGenXisalsopresentindrinkingwaterandaccumulatesintheblood(NorthCarolinaDepartmentofEnvironmentalQuality).TheexactmechanismofthediseasescausedbyPFASisstillunknown.Researchhasyettodisclosealloftheeffectsthesechemicalsmayhaveonthebody,andhowtheyarecaused.Onepotentialmechanismofdiseasemaybethroughdisruptionofbeneficialbacteriainthegut.Thehumanbodyisknowntohavealargevarietyofflora,particularlyconsistingofavarietyofbacteria,thathavebeenshowntoplayacrucialroleinhumanhealth,anddamagetothemicrobiomehasbeenlinkedtocancer,liverdisease,diabetes,andmore(Jandhyalaetal.,2015)(Sekirovetal.,2010).Lactobacillusacidophilusisaspeciesofbacteriafoundinthehumangastrointestinaltract.Thisspecieshasbeenshowntohavemajorbeneficialrolesinhumanhealth:Lactobacillushasbeenshowntoprotectagainstliverdisease,andincreasednumbersofthebacteriumarealsorelatedtoaloweredriskofcolorectalcancer(Ritzeetal.,2014)(MooreandMoore,1995)(GuarnerandMalagelada,2003).ColorectalcanceristhethirdleadingcauseofcancerdeathsintheUnitedStates,andisaffectingeverincreasingamountsofyoungpeopleintheUnitedStates(Siegel,Jemal,andWard,2009).Becauseofthisconnection,killingthesebeneficialbacteriacouldbedetrimentaltotheoverallhealthoftheUSpopulation(Zhong,Zhang,andCovas,2014).Despitethesmallamountsofcurrentliteratureonthesubject,PFAShavebeenshowntodisruptthemembranebilayerofbacteria,andathighenoughconcentrationshavebeenshowntodamageandevenkillthebacteria(Fitzgeraldetal.,2018).BecausePFASarefoundinwater,havebeeningestedbyhumans,andareevenfoundintheblood,itisentirelypossiblethatthepresenceofPFASintheGItractcaninterferewithorevenkillbacteriainthegut,includingthebeneficialLactobacillus.ThedeathofLactobacillusbacteriamaythusbeacausalfactorindiseasessuchascolorectalcancerandliverdisease.ThepurposeofthisexperimentistoinvestigatetherelationshipbetweenPFASandhumanhealthbydeterminingtheeffectofPFASonthebeneficialbacteriaofthegut.ThreePFASwillbetested-thecommonlyfoundPFOAandPFOS,andthenewGenXchemical.ItishypothesizedthatallPFAStested,includingthe“safe”GenX,willhavesomeinhibitoryeffectsonLactobacillusacidophilus,demonstratingapossiblemechanismofdiseaseinthehumanbody.Methods:TheeffectofPFASonLactobacillusacidophiluswastestedintwoways.OneofthesetestsmeasuredthezonesofinhibitioncreatedbytreatingLactobacillusacidophiluswithmultipleconcentrationsofPFAS.Whenapetridishofbacteriaistreatedwithasmalldisccoveredinaninhibitorychemicalandallowedtoincubate,acircleofnobacterialgrowthwillappeararoundthedisc-thisareaismeasuredandiscalledazoneofinhibition.Thelargerazoneofinhibitionis,thestrongertheinhibitoryeffectwas.Theothertestperformedwasaspectrophotometricassaythatallowedforthegrowthofthebacteriatobemeasured.Eachbacterialspecieshasacharacteristic“growthcurve”thatcanbegraphedwhenmeasuringabsorbanceofabacterialsampleoveralongperiodoftime.Ifthebacteriaistreatedwithaninhibitorychemical,thischaracteristicgrowthcurvewillchange.Escherichiacoliwasusedasacontrol,asithasdemonstratedsensitivitytoPFASinpreviousresearch.Whencompared,thedatawillshowifLactobacillusacidophilusmaybeevenmoresensitivetoPFASthanE.coli.Zonesofinhibition:50PetridisheswereinoculatedwitheitherLactobacillusacidophilusorEschericiacoli.Diffusingdiscswereplacedonthecenterofeachplate,andeachofthediscswastreatedwith40ulofeitherahighPFOAconcentration,lowPFOAconcentration,highPFOSconcentration,lowPFOSconcentration,highGenXconcentration,lowGenXconcentration,ornoPFASatall.Thiswasrepeatedintriplicate,sothat3ofeachtreatmentgroupforeachbacteriacouldbemeasured.Thesepetridisheswereincubatedat37degreescelsiusforfournights,andanyzonesofinhibitionweremeasured.Spectrophotometricassay:Six50mLtubesofliquidgrowthmediaweretreatedwitheitherahighPFOAconcentration,lowPFOAconcentration,highPFOSconcentration,lowPFOSconcentration,highGenXconcentration,orlowGenXconcentration.A4mLaliquotwasremovedfromeachofthelargerstocksandtransferredintoaseparate5mLtesttube,foratotalof6tubes.Oneadditionaltubeof4mLuntreatedliquidgrowthmediawasalsoprepared.Intoeachofthesetubes,100ulLactobacillusacidophilusculturewasplaced.Thisprocesswasrepeatedtwomoretimes,fortriplicatemeasurements.Theaboveprocedurewasfollowed,butinsteadofplacing100ulLactobacillusintothetubes,50ulE.coliwasusedinstead,toallowforacontroltocompareLactobacillusto.Thiscreatedatotalof42tubes.All42tubeswereplacedintoawarmwaterbathat37degreescelsius,andshakenovernight.Beginningthenextmorning,theabsorbancelevelsofthetubesweremeasuredusingaspectrophotometerat660nanometerswavelength.Thiswasperformedapproximatelyeachhourfor8hours.TheabsorbancedatawasusedtocreateagrowthcurveforthenormalbacteriaandeachofthedifferentPFASgroups.Dataanalysis:AnANOVAwillbeperformedonthezoneofinhibitiondatatodeterminewhichPFAShadthemosteffect,atwhatconcentration,andwhichbacteriawasmostaffected.Theaveragegrowthcurveforeachtreatmentgroupwillbeplottedandcomparedtothe“normal”growthcurvetodeterminetheeffectsofdifferentconcentrationsofPFOA,PFOS,andGenXonthebacteriaovertime.

RebeccaWeipert
DoesChangeinLocationAffecttheFrequencyorSeverityofAllergies?

Theobjectiveofmystudyistodeterminewhetherthereisacorrelationbetweenindividualsthathavemovedlocationsfromchildhoodandadulthoodandhaveincreasedsensitivitytoallergens.Allergiesarecausedbytheimmunesystem’sresponsetoallergens.Allergensareonetypeofantigenwhichpromoteantibodyproductioninthebody.Allergenspromoteaspecificantibodytoproduce,ImmunoglobinE(IgE)antibodies.InthepresenceofIgEantibodies,allergysymptomsoccur.TheseallergysymptomsarealsoclassifiedasTypeIHypersensitivityreactions.Inthisstudy,theallergensthatarebeingfocusedonaresubstancesthatarebothforeignandharmlesstothebodyandcausesymptomsofhayfever.Hayfeversymptomsincludeanycombinationofthefollowing:runny/stuffynose,itchy/wateryeyes,inflammationofairway,headache,fatigueandphlegm.Themostcommontypesofallergenstocausethesesymptomsarepollen,dust,moldanddander.Mastcells,basophilsandeosinophilsarealltypesofwhitebloodcellsthatareactivatedfromtheproductionofIgEantibodies.Increasednumbersofeosinophilscorrelatewiththepresenceofallergies;therefore,youareabletocounteosinophilsinbloodsamplesanddetectthepresenceorabsenceofallergies.Astudyconductedin2012showedthatchildrenlivinginurbancitieshadahigherprevalenceofallergicdiseasesthanchildrenwholivedinruraltownsandvillages.Thechildrenlivingintheurbanareasalsoreportedthattheyhadexperiencedmorefrequentandsevereallergiesthanthechildrenlivingintheruralareas.Toconductthisstudy,aminimumofthirtyLSSUstudentparticipantswillbeneeded.Theparticipantswillbeassignedintogroupsaccordingtotheirchildhoodresidences.HalfoftheparticipantswillbeinagroupwiththeirresidencebeingSaultSte.Marie(SSM)andcurrentlyresidethere.TheotherhalfwillbeinanothergroupwiththeirchildhoodresidencesfromalocationotherthanSSMbutlivetherecurrently.Thisgroups’childhoodresidencesmustnotbewithin115-mileradiusofSSM.Questionnaireswillbeprovidedtoeachparticipantpertainingtopasthometowninformationandfrequencyand/orseverityofallergysymptomssincebeinginSSM.FingerpricksandHemoCuewhitebloodcellanalyzerwillbecarriedoutandusedinordertoobtainbloodeosinophilcountsineachparticipant.

SydneyZuke
ExaminingtheCorrelationBetweenVariationsoftheACTN3GeneandAthleticPredispositioninDivisionIIStudent-AthletesatLakeSuperiorStateUniversity

Inthescientificcommunity,ithasbeenwidelyacceptedthateliteathleticperformanceisaresultofacombinationofenvironmentalfactorsandgeneticmakeup(Georgiades,2017).Althougheliteathleticperformanceisdependentuponmanydifferentgenes,thisresearchwillexamineaspecificgene,alpha-actinin-3(ACTN3),alsoknownasthe“sprinter”or“speed”gene,anditsgenotypes(Pickering&Kiely,2017;DelCosaetal.,2019).Thisgeneisofparticularinterestbecauseitencodesthealpha-actinin-3proteinwhichisresponsibleforgeneratingfast-twitchmusclefibers.Fast-twitchmusclefibersareresponsibleforgeneratinghighamountsofforcequickly.Therefore,thevariantformoftheACTN3geneiscommonineliteathletesthatparticipateinsportsthatrequireexplosivepower,suchassprinting(Ostrander,Huson&Ostrander,2019).TheACTN3genotypewillbeexaminedtodetermineitspresenceinDivisionIIstudent-athletesatLakeSuperiorStateUniversityinordertounderstandifthosestudent-athleteswerepredisposedtoathleticsuccessbecauseoftheACTN3geneoriftheirsuccesswasdeterminedprimarilythrougheffort.Thus,weareexaminingwhethernaturepredisposesDivisionIIathletesthesameaswitnessedineliteathletes,orifnurtureandtrainingplayalargerroleinsuccess.Thisstudywilltakesamplesfromathletesofbothanaerobic(powerandspeed)prevalentsportstudent-athletesandaerobic(endurance)prevalentsportstudent-athletestodeterminethedegreeofprevalenceoftheACTN3genotype.TheinformationwillbeusedtodetermineifthereisacorrelationbetweensporttypeandthepresenceoftheACTN3genotypeinthispopulation.ThiswillbeaccomplishedthroughthecollectionofDNAsamplesthroughcheekswabbingandsubsequentanalysis.Normanetal.(2014)conductedsimilarmethodology.PurposeStatement:ThepurposeofthisresearchstudyistodetermineifthereisasignificantcorrelationbetweenthepresenceofACTN3genotypeandathleticpredispositioninDivisionIIstudent-athletesatLakeSuperiorStateUniversity.Relationtofieldofstudy:Nature,orgeneticpredisposition,hasbeenproventobeaprecursortoathleticsuccessattheelitelevel(Pickering&Kiely,2017)Asthecompetitionbecomesstronger,theathletemusthavenurturedtheirathletictalents,buttheymustalsohavesimilargeneticpredispositionsinordertotrulycompete(Ostrander,Huson&Ostrander,2019).Thus,weseeathletesselectingonesporttocompeteinattheelitelevel.Thisisnottrueatlowerlevels.Atthehighschoollevel,itiscommonforathletestoplay2,3oreven4sports(Belletal.,2016).Someoftheseathletesmaynotplaysportsatthecollegiatelevel.However,forthosethatdo,theymustrisetoanewlevelofcompetition.ItisnotknownifthelevelofcompetitionattheDivisionIIlevelis‘elite’enoughinwhichgeneticsmustplayaroleinorderforathleticsuccess.Thus,thisstudyaddstothefieldofkinesiologybybeginningtoanswerthequestionregardingnatureversusnurtureattheDivisionIIlevelofcompetition.Theanswertothisquestionhasmuchtoofferintermsoftrainingmethodologyandrecruitmentofstudent-athletes.Methodology:UponapprovaloftheHSIRBcommittee,student-athleteswhoparticipateinDivisionIIathleticsatLakeSuperiorStateUniversitywillbeaskedtoparticipateonavolunteerbasis.Followingtheinformedconsentprocess,student-athletesfromanaerobicandaerobicbasedsports(tennis,basketball,volleyball,cross-country,trackandfieldandgolf)willbeaskedtosetupatimeforDNAcollectionthroughswabs.Noprocedureswillbeperformeduntilinformedconsentisprovidedtotheparticipants.Student-athleteswillmeettheprimaryandfacultyresearcherinthegeneticslabandcompleteashortdemographicsurveytoidentifygenderandsport.Numberswillbeusedinplaceofidentifyinginformation.Followingthedemographicsurvey,theprimaryresearcherwillconductacheeksalivaDNAswab.FollowingtheDNAswab,student-athleteparticipationinthisstudywillbecomplete.TheprimaryresearcherwillimmediatelypreparetheswabforDNAanalysistodeterminethepresenceoftheACTN3genotype.

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Spring2020GrantAwardees

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KassidyHadix

IwillbelookingforthefrequencyoftwocommongenemutationsintheEasternUpperPeninsulaCaucasianpopulation.ThegenesofinterestareFactorV(responsibleforbloodclotting)andmethylenetetrahydrofolatereductase(MTHFR,whichregulateshomocysteinebuildup).Thesemutationsareimportantbecausetheycanleadtoseriousconditions,suchasabnormalbloodclotsandmiscarriages. DNAsampleswillbeobtainedfromnon-invasivecheekswabs;amplifiedusingPCRandquantifiedusingthenewgenefragmentanalyzerintheSchoolofScienceandMedicineatLSSU.WewillcomparetheEUP’sprevalenceofthesemutationstothatoftheoverallU.S.population.Knowingthefrequencyofthesetwomutationscanhelptoeducatehealthcareprofessionalsandthegeneralpublicabouttherisksassociatedwiththeseconditions,sothatpeoplecanbetterregulatetheirhealth.

BreannaKemppainen
Chemicalcleanersareusedtocleansurfacesinhouseholds,hospitals,andpublicplacestoprotectusfromillness.Mostcleanersuseaspray-then-wipemethodbutitisunclearhowthephysicalactionofwipingcontributestoremovingbacteria.Somecleanersmaynotbeactuallykillingbacteria,butfrictionfromwipingmaybeenoughtophysicallyremovethem.Inthisstudywewillclarifythecontributionthatfrictionmakestocleaningsurfacesbycountingthenumberofbacteriaremovedfromasurfacewhenusingdrymaterialsversustheamountremovedwhenusingthesamematerialwetwithwateroracleaner.

SophieMcConkey

Theintentofthisresearchprojectistoenableamobilerobottoaccepthumanassistanceforthepurposeofdeviatingfromitsautonomouslygeneratedpath,thusreachingitsdesireddestination,whileavoidingobstructions.TheresultsofthisresearchwillenableAutomatedGuidedVehiclestoovercomethechallengeofnavigationwithindynamicenvironments.TheteamplanstouseaTurtleBot3WafflePi,equippedwithaColorSensorDistanceMeasuringSensor,andGyroSensor,an8Megapixelcamera,andaNvidiaJetsonTX2computerboardtoaccomplishthistask.

RebekkaRanta

Identifyingandunderstandingtheeffectthatperceivedstressandresiliencelevelshaveonanindividual’sperformancewhenpresentedwithanacute-stressinducingtaskisveryimportant. Foruniversitystudents,manyarefrequentlyfacedwithnegativesituationsthatarenotalwaysdirectlyrelatedtoacademics.Byidentifyingsuch,thewayanindividualperformsunderpressurecanbebetterunderstood.Participantswereevaluatedontheirstressandresilienceperceptions,theirperformanceonthestress-inducingtask,andtheirheartratelevels.Researchsurroundingthistopicisofvaluebecauseitprovidesinformationthatcouldimprovethehealth,well-being,andretentionofuniversitystudents.

GeorgeSchaffer

Iamresearchinghowlungfunctionmayinfluencesymptomsofpanic,suchasshortnessofbreath,aracingheartbeat,feelingfaint,orshaking.Imeasuredlungfunctionusingaspirometer,asmallhandhelddevicethatmeasuresexhalationspeedandlungcapacity.Ialsoaskedparticipantstofilloutademographicsurveyandapanicsurveydetailingtheirhistoryofpanicattacks,thespecificsymptomstheyhadifthey’dexperiencedapanicattack,andwhetherornottheyhadexperiencedapanicattackrecently.

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Fall2019GrantAwardees

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BrendanLukomski

ComparingrhizosphericdifferencesbetweenyounggrowthandoldgrowthEasternWhite Pine(Pinusstrobus)

InthisstudythegoalwillbetodeterminedifferencesinrhizosphericsoilmicrobialcommunitiesbetweendifferingagedstandsofEasternWhitePine(Pinusstrobus). Soilsampleshavebeencollectedat3locations;asitenearTahquamenonStatePark,alocationjustsouthwestofSaultSte.Marie,MI,andasitenearCopperHarbor,MI. TheextractedmicrobialDNAfromthesoilsamplesarethenanalyzedformicrobialcommunityabundanceaswellasdiversitywithinthecommunity. ThiswilldeterminewhetherornottheEasternwhitepineindividualsareselectingcertainmicrobialcommunitycharacteristicsthroughouttheirlifetime.

BriannaRegan
Theimpactsofenrichmentontheproblem-solvingabilitiesofrats(Rattusnorvegicus)

Thephysical,psychological,andoverallwell-beingofanimalscanbenegativelyimpactedbycaptivity.Enrichmentisamethodofkeepingcaptivespeciesengagedbygivingthemactivitiestopartakeinthroughouttheday.Enrichmentdevicescouldincludetoys,noveltyobjects,food,sensoryactivities,andmuchmore. Myprojectwillutilizeratstoinvestigatetheimpactsenrichmentcanhaveonproblem-solvingability.Therewillbeacontrolandatreatmentgroup.Theratsinthecontrolgroupwillnotbeexposedtoenrichment,whiletheratsinthetreatmentgroupwill.Eachratwillbeevaluatedbyrecordingthetimeittakesthemtocompleteamazebeforeandaftertheexperimenttakesplace.Thisdatawillbeanalyzedtolookfordifferencesbetweentheratsthatwereexposedtoenrichmentandtheratsthatwerenottoassesshowproblem-solvingabilityisimpactedbyenrichment.[CP_CALCULATED_FIELDS_RESULT]

LucasSheppard

“EffectsofsalinityonserumcortisolinAtlanticsalmon”

ThisprojectwilllookintosafehandlingtechniquesofAtlanticsalmon(Salmosalar)inregardtoperforminga“Dip”intoasaltbathforvaccinations.Salmonwillberandomlyassignedintothreesalttestgroupswithafurtherlevelofvariationbeinglengthoftest.Bloodwastakenpostdifferentsalinityexposureinordertodetermineasafeconcentrationofsalttousefora“Dip”aswellaslengthoftime.TotalbloodcellcountandplasmacortisollevelswereanalyzedundermicroscopeandviaELISAtestrespectively.ANOVAtestwasusedfordataanalysistofindadifferencebetweenthetests:serumcortisolandsalinitytreatment

VictoriaSize
TheEffectsofPFASontheDevelopmentandMitochondrialFunctioninZebrafishOva

RenéeResendesofWindsor,ONtestedtheeffectsofPFAS(Per-andPolyfluoroalkyl
Substances)onthedevelopmentandmitochondrialfunctionofzebrafishova.Thetwomost
commonformsofPFASarePFOAandPFOS,whicharepersistentintheenvironmentandof
commonexposuretohumans.Thestudyaimedtodetermineiftherewasacorrelationbetween
changesinembryonicdevelopmentandenergyproduction,asaresultofexposuretoPFOA,
PFOS,andincombination.HerresultsshowedthatPFOAandPFOSincombinationaffected
embryonicdevelopment,causedchangesinenergyregulation,andwithintheseconditionseven
resultedinembryodeathbefore14daysold.ResearchofPFASisofvaluebecauseofthe
potentialharmthesechemicalshaveonreproductiveanddevelopmentalprocessesandoverall
adversehumanhealtheffects.

JessicaThompson

StresslevelsrelatedtoAcetaminophenusageinsocialconformity

Recently,somenewlyfoundsideeffectsofacetaminophen,theactiveingredientinTylenol,haveshownsuppressedemotionaloutput.Thisstudywasdesignedtousesocialconformityasamediumtomonitoranybehavior,mood,orphysicalchangesacetaminophenmayhaveonthosewhoconsumeit.Atotalof48participantswereevaluatedduringasmallgroupactivityontheircortisollevels,moodstates,bloodpressure/pulse,andbehavioralchanges.Thedatagainedwillbeanalyzedtodetermineifacetaminophenisdecreasingphysiologicalstressreactionandmoodchanges,andifthesuppressionofstressmayresultinabehavioralchange.