ACURRICULUMforHIGHSCHOOL SCIENCEEDUCATION:

MICROBIALFUELCELLS: ALivingBattery

BrettBarron
HighSchoolScienceTeacher, HazelwoodCentralHighSchool,St.Louis,MO Email:bbarron@hazelwoodschools.org

MiriamRosenbaum,PhD

ResearchAssociateMicrobialFuelCellResearch DepartmentofBiologicalandEnvironmentalEngineering,CornellUniversity Email:mr625@cornell.edu

PhyllisBalcerzak,PhD

ScienceEducationSpecialist ScienceOutreach,WashingtonUniversityinSt.Louis, Email:pbalcerz@biology2.wustl.edu

LargusT.Angenent,PhD

AssociateProfessor BiologicalandEnvironmentalEngineering, CornellUniversity 214RileyRobbHall,Ithaca,NY14853 Email:la249@cornell.edu Phone:6072552480 Weblink:http://angenent.bee.cornell.edu/

Thisworkwasmadepossiblethrougha NSFCAREERgranttoLarsAngenent(#0939882).
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MICROBIALFUELCELLS:ALivingBattery

MOTIVATION

Microbialelectrochemistryprovidesauniquemethodforpromotinganinterdisciplinary approachtoteachingscience.Mostsciencecurriculatoday,adaptingtothegrowingdemand to align course content to state standards, veer away from the integration of biology, chemistry, and physics. Many times, this does a great disservice to our students. The life processesstudiedinthesecoursesdefyourattemptstosimplifyandcategorizethemtheyare infinitely complex, and often these complex problems require the integration of biology, chemistryandphysicstofullyexplainthem.Inatypicalhighschoolsciencecourse,thestudent acquiresfragmentedideas(factoids)aboutthesubjectofstudy,andwillvaryintheamountof information they have acquired. Frequently, the average student has great difficulty in conceptualizing the patterns and relationships between the various science disciplines. For example, students can master the basics of oxidationreduction from their introductory chemistry class, yet when called upon to be familiar with the metabolic pathways of cellular respirationinabiochemistryunit,failtorecognizehowredoxchemistrygovernsthebiological processes. Ourgoalisfirsttodemonstratehowanintegratedcomplexsystem(e.g.,amicrobialfuel cell)canbeusedtosuccessfullyteachscience(biology/chemistry/physics),andthenprogressto how science can be used to solve complex problems (e.g., energyneutral wastewater treatment). Essentially, we hope to provide opportunities for students to transform factual knowledge into usable information for problem solving. Often, this type of investigation not onlystimulatesthecuriosityofstudentsinbiotechnologyanditsrelationtothebasicsciences, butalsoencouragesamoregeneralflexibilityofscientificthinkingintheirinterpretations.

Demonstrationsofmicrobialelectricitydevicesoftencapturetheattentionofhigh school audiences, and the intention of this curriculum is to provide the tools for instruction and some relatively simple experiments that can be performed in a high school laboratory. Some oversimplification of the basic biochemical and microbiologicalprocessesisperhapsinevitable,andwherenecessaryreferences to standard texts should be tailored to the needs of your students. Where applicable,attentionwillbedrawntotherationaleforeachactivitybydescribing how the students can link their understanding of each process to the understandingofafunctionalMFC.

INTRODUCTION

Asaresultofglobalpopulationgrowthandcumulativeindustrializationprocesses,two tremendouschallengesinourworldtodayarethegrowingdemandforenergy,andecologically andenvironmentallysoundmethodsforwastedisposal.While,arguably,wepossessadequate methods for waste treatment and disposal, they are quite energyintensive processes. Some currentresearch,especiallyintomicrobialfuelcells,asksthequestionifwastewatertreatment can be transformed into energy producing processes. As fossil fuels are exhausted, could smaller,tailoredsolutionsdesignedforindividualenergydemandscometoreplacethehuge, centralized power plant? In our evolving energy conscious environment, could wastes ever cometoplayadistinguishedroleasaresourcefortheprovisionofelectricalpower?Theseare questions researchers in the field of microbial fuel cells hope to answer. Wastes, contrary to conventional wisdom, are not useless as the name may imply rather, they are valuable energysources.Municipalwastewaterpossessesanorganicloadof~200mg/LCOD,andwhile thisisnotenoughrawenergyforwastewatertoeverbeamajorproducingsource,thereare some that argue the treatment could become an energyneutral process rather than energy consuming. Special industrial wastewaters, such as from the food industry (dairy, brewery, sugar industry), being much richer in organic content, may potentially hold an even more promisingenergyproductionfuture. Anovelapproachtofacilitatedirectenergyproductionfromwastewateristhemicrobial fuel cell. The first studies into the application of direct transformation of chemical into electrical energy by the exploitation of microbial processes arose during the 1970s. Mainly becauseoftheabundanceoffossiloilandgas,itreceivedonlytemporaryattention.Itwasnot untilthe1990s,wheninterestinsustainableandrenewableenergysourcesexpanded,thatthe ideawasrevivedandtheresearchintensified.Althoughmuchworkremainstobedoneatthe research level to unwrap the biochemistry of the electrical activity of microorganisms, recent studiesofmicrobialfuelcellshavegreatlyadvancedourunderstandingofmicrobialelectricity generation.Additionally,studyingthesemicrobialfuelcellscanleadtoadeeperunderstanding of electron transfer processes in general and the application of this knowledge to other alternativefuelpossibilities.ForinformationontheworldwidestatusofMFCresearch,consult thewebpageoftheMFCresearchcommunity:www.microbialfuelcell.org.

Chapter1:WhatisaBattery?

BATTERYBASICS Havethestudentsgivetheirdefinitionofabattery.(NOTE:Thepubliccallsitabattery, buttheindustryreferstoitasacell.) A battery is an electrochemical device that contains two or more power cells connected electricallysothatchemicalenergycanbeconvertedintoelectricity.Simplystated,abattery powers products that require electricity to work. They are useful because they allow us to transportelectricityanduseproductsinlocationswheretherearenoelectricaloutlets,suchas beaches,sportingevents,picnics,etc. Inabattery,eachcellthatstorestheelectricalenergyinachemicalstatehastwoelectrodes thatreactwiththechemicalandeachothertoreleaseenergy.Thebatterystwometalendsare called terminals. Usually one terminal is flat (negative end) and the other is buttonshaped (positiveend).InatypicalCarbonZincbattery,thepositiveelectrodeisacarbonrodandthe negativeelectrodeisthezinccase.Thecaseisimportantalsobecauseitkeepsthechemicals fromleakingout. Lookfortheterminalsonabattery.Howaretheymarked? Theendsaremarkedwitha+and. MFCswillalsohavepositiveandnegativeterminals.OurspecificlaboratorymodelMFC, instead of having just two terminals at opposite ends, will have three terminals (two negativeandonepositive)allcomingoutofthetopoftheMFC.Theterminalswillbethe conductive carbon electrodes (rods) that emerge from the two anolytic (negative) chambers and the catholytic (positive) chamber. The two negative electrodes will be linkedbyalligatorclipsandcopperwire,essentiallycreatingonenegativeterminalfrom thetwo.

Since our microbial fuel cell effectively acts as a battery, it is beneficial to begin by demonstratinghowitfunctionsasabattery.WhenstudentsconceptualizethattheMFCis actingasabattery,theycaninternalizethatalloftheprocessesthatoccurwithinatypical dry cell battery are also occurring within the MFC. The topics begin with a review of the designandoperationofabasicbatteryandthenapplythoseconceptstotheMFC.Partsof thistopicmaybeunnecessaryifthestudentshavepriorknowledgeofbatteriesandcircuits gainedatthemiddleschoolorintroductoryphysicalsciencelevel.

HowdoesunderstandingbatterieshelpstudentsunderstandMFCs?

Eachbatteryconsistsoffourmainparts: a)PositiveElectrode(Cathode)theelectronacceptor. b)NegativeElectrode(Anode)theelectrondonor. c)Electrolyteapastelikesubstanceorsolutionthatcontainschargedparticlesthat canmoveorconductanelectriccurrent. d)Separatormaterialthatprovidesseparationofthe(+)fromthe()chargesand insulation. The electrochemical reaction that fuels the battery occurs at the interface of positive and negativeelectrodesandtheelectrolytes.Electricitycreatedbyabatteryconsistsofastreamof tinyinvisibleparticlescalledelectronsflowingfromonemetalendofthebatterytotheother metal end, just like a liquid. The path it follows is called a circuit. When electricity flows in a circuit,itiscalledcurrent.Electricityonlyflowswhenitcangofromoneterminaltoanother. The electrons must have a pathway or circuit to follow from the positive to the negative electrode. When the circuit is complete, electricity flows from an area of low (more negative) electrical potentialtooneofhigher(lessnegativeorevenpositive)potential.Thedifferenceinelectrical potentialmakestheelectricitymove.Electricitycanflowthroughsomethings,butnotthrough others.Materialsthatallowelectricitytoflowthroughitarecalledconductors.Metalsusually makegoodconductors.Ifelectricitycannotflowthroughthematerial,itiscalledaninsulator. Whatiselectricity? Electricity or electric current is a movement or flow of electrically charged particles,calledelectrons. Canyounamesomematerialsthatmakegoodconductorsandinsulators? Good conductors include aluminum, carbon, copper, salt water, and zinc. Insulatorsincludedrysalt,glass,plastic,andwood. Theelectricalsymbolforabatteryis: The longer line indicates positive and the shorter line indicate negative. All batteries have a strengthindicatedbyanumberfollowedbytheletterV.TheVstandsforvolts.Thevoltis an electrical unit that measures the potential difference between two points in an electrical circuit.Thepotentialdifferencebetweentwopointsistheamountofworkthatwouldneedto bedoneonaunitofelectricchargetomoveitfromonepointtotheotheragainstanelectric field.

Thevoltagenumberforabatterygivesarelativemeasureofhowhardtheelectronsarebeing pulledthroughthecircuitfromanareaoflowelectricalpotential(moreelectronrich)tooneof higher potential. For instance, a 1.5 V battery contains a single cell whereas a 9 V battery containssixcells,withthecurrentmovingthroughallsixcells.

Whatdothenumbersonabatterymean?Howdotheyrelatetothestrengthofthe battery?Canyousuggesthowvoltagecouldbeincreased? The numbers indicate voltage. Batteries with higher numbers have a higher strength.Youcanmakeavoltagestrongerbylinkingseveralbatteries.
Activity1.1(Demonstration)LightItUp Showthebrightnessofabulbresultingfromoneandtwobatteriesaddedinaseries circuit.Then,useamultimeterorvoltmetertoshowhowthevoltageisadditive.

PRIMARYANDSECONDARYCELLS Themostcommonhouseholdbatteryisadrycellbattery.Adrycellbatteryischaracterizedby apasty,lowmoistureelectrolyte.Onekindofdrycellisaprimarycellbattery.Thesebatteries automaticallyconvertchemicalenergyintoelectricalenergy.ThiskindofbatteryCANNOTbe rechargedbecauseithasadefinitefuelstock.Itisdesignedtobeusedonce. After the chemicals in the electrolyte solution (that transmit the electric currents) have been used up, the energy is no longer available and the battery is said to be exhausted, used, or dead,andisthendiscarded.Anexampleofsuchaprimarycellbatterywouldbethealkaline battery,usedinmostremotecontrols.Theanodeofthisbatteryisamixtureofpowderedzinc and a concentrated solution of potassium hydroxide. The cathode is a mixture of solid manganese (IV) oxide and graphite. The anode and cathode chambers are separated by a porousdivider.Themostcommonsizesofalkalinebatteriesaretheroundcells,suchasAAA, AA,C,andD(Figure1.1).

Figure1.1.TypicalalkalinebatteriesdepictingD,C,AA,andAAAsizes.

AsecondarycellbatteryCANberechargedandusedrepeatedly.Thedischargedenergycanbe restoredbyforcingelectronstoflowintheoppositedirectionbyutilizinganexternalelectrical energysource.Anexampleofasecondarybatterythatcanbeusedforanumberofyearsisthe leadacidbatteryorleadstoragecellfoundinautomobiles(Figure1.2).Themainfunctionofa car battery is to store electrical energy for starting the car and operating electrical devices whenthecarisnotrunning.Bothelectrodesintheleadstoragecellcontainaleadgrid.The anodesareimpregnatedwithaspongyleadmetal,whileinthecathode,thegridispackedwith redbrown,lead(IV)oxide.Adilutesulfuricacidsolutionservesastheelectrolyte.Thebattery isconstantlybeingdischargedeverytimetheengineisstarted.Duringthischemicalreaction, sulfuricacidisturnedintowaterandbothelectrodesareconvertedtoleadsulfate.Whilethe carisbeingdriven,electricalenergygeneratedbythealternatorreversesthedecomposition, andthebatteryiscontinuouslyrecharged.

Figure1.2.Atypicalautomotiveleadacidbattery

CathodeReaction(reduction): PbSO4(s)+5H20(l)PbO2(s)+3H3O+(aq)+HSO4(aq)+2e

AnodeReaction(oxidation): PbSO4(s)+H3O+(aq)+2ePb(s)+HSO4(aq)+H2O(l)

Oneofthemostrecentlydevelopedrechargeablebatteriesisthelithiumionbatterycommonly usedincellphonesandlaptopcomputers(Figure1.3).TheanodeconsistsofLi+ionsthathave been inserted reversibly into layers of graphite. The cathode contains lithium cobalt oxide (LiCoO2).Whenthecelldischarges,Li+ ionsspontaneouslymigratefromthegraphiteanodeto the cathode, andelectrons flow through the external circuit. When the battery is recharged, cobaltionsareoxidizedandLi+ionsmovebackintothegraphite.

 Figure1.3.Atypicalcellphonelithiumionbattery Havestudentsresearchvarioustypesofsecondarycellbatteriesandreportonsuchtopics ashowitworks,thelifespanofthebatteries,usesofthebatteries,andcost. MFCscannotberechargedbyreversingtheflowofelectrons,butsincetheyareatype of fuel cell, they will run indefinitely as long as a fuel feed is constantly supplied. This ensures the bacteria are constantly metabolizing glucose to produce electricity. The potassium ferricyanide solution can be recirculated in the cathode chamber for an extendedperiodoftimeaswell,butshouldbereplacedregularly. ELECTRICALCIRCUITS An electric circuit is an interconnecting path, external to the battery, which allows charge to flowbetweenthepositiveandnegativeterminalsofthebattery.Asimplecircuitmayconsistof asinglestrandofmetalwirelinkingtheterminals.However,amorerealisticcircuitpossesses multiple branch points so that charge can take many different paths between the two terminals. Although there can be many different paths through the external circuit that the charge could take, the electrical energy that the charge acquires in making this journey is alwaysthesame.Since,whenanalyzingelectricalcircuits,weareprimarilyinterestedinenergy (i.e., in the transformation of the chemical energy of the battery into heat energy in some electric heating elements, or mechanical energy in some electric motors, etc.), it follows that the property of a battery, which primarily concerns us, is its voltage. Recall that the voltage number V for a battery gives a relative measure of how hard the electrons are being pulled throughthecircuit. The rate at which charge flows out of the positive terminal is termed the electric current flowingoutofthebattery.Likewise,therateatwhichchargeflowsintothenegativeterminal istermedthecurrentflowingintothebattery.Ofcourse,thesetwocurrentsmustbethesame otherwise charge would build up in either the battery or the circuit. Electric current, representedbytheletterI,ismeasuredinunitsofamperesoramps,abbreviatedA.

Though,inactuality,thecurrentiscarriedbynegativecharges(i.e.,byelectrons)flowinginthe opposite direction, conventionally, the direction of the current is taken to be the direction positive charges would have to move to account for the flow of charge. The current at all points in the external circuit must remain constant over time. We call this type of circuit a directcurrentorDCcircuitbecausethecurrentalwaysflowsinthesamedirection.Thereisa secondtypeofcircuit,whichiscalledanalternatingcurrentorACcircuit,inwhichthecurrent periodicallyswitchesdirection. A simple circuit can be described as being somewhat analogous to a small ski resort. The charges flowing around the external circuit are like people skiing down the skislope. The charges flow down a gradient of electric potential just as the people ski down a gradient of gravitational potential. Note that the good skiers who ski directly down the slope acquire exactlythesamegravitationalenergyasthepoorskierswhoskifromsidetoside.Inbothcases, the total acquired energy depends only on the difference in height between the top and bottom of the slope. Likewise, charges flowing around an external circuit acquire the same electricalenergynomatterwhatroutetheytakebecausetheacquiredenergyonlydependson thepotentialdifferencebetweenthetwoterminalsofthebattery.Oncethepeopleinourski resortreachthebottomoftheslope,theymustbeliftedtothetopinaskiliftbeforetheycan ski down it again. Thus, the skilift in our resort plays an analogous role to the battery in our circuit.Ofcourse,theskiliftmustexpendnongravitationalenergytoliftskierstothetopofthe slope, in just the same manner as the battery must expend nonelectrical energy to move chargesupapotentialgradient.Iftheskiliftrunsoutofenergy,thenthecirculationofskiersin the resort rapidly stops. Likewise, if the battery runs out of energy (i.e., if the battery ``runs down''),thenthecurrentintheexternalcircuitstopsflowing. OHMSLAW Consider,again,asimplecircuitinwhichasteadycurrentIflowsthroughasingleconducting wire connecting the positive and negative terminals of a battery of voltage V. What is the relationship between the current I flowing in the wire and the potential difference V applied across the two ends of the wire by the battery? If we were to investigate this relationship experimentally,wewouldquicklyconcludethattheelectricalcurrentIisdirectlyproportional tothepotentialdifferenceV.Inotherwords,

V=IR

wheretheconstantofproportionalityRistermedthe(electrical)resistanceofthewire.The above formula is called Ohm's law after its originator, the early nineteenth century German physicistGeorgSimonOhm.Theunitofelectricalresistanceistheohm(). Goodconductorsofelectricity(i.e.,copper,silver,aluminium,andmostothermetals)possess nonzero electrical resistances. However, these resistances are generally so small that if we were to connect the terminals of a battery together using a wire fashioned out of a good conductor,thenthecurrent,whichwouldflowinthewire,accordingtoOhm'slaw,wouldbeso 9

largethatitwoulddamageboththewireandthebattery.Weusuallycallsuchacircuitashort circuit. To prevent excessively large currents from flowing, conventional electric circuits containcomponentscalledresistors,whoseelectricalresistanceismanyordersofmagnitude greaterthanthatoftheconductingwiresinthecircuit.WhenweapplyOhm'slaw,V=IR,toa circuit,weusuallyonlycountthenetresistanceRofalltheresistorsinthecircuit,andneglect theresistancesoftheinterconnectingwires.Thismeansthatallofthemajordropsinelectric potential, as we travel around the circuit from one terminal of the battery to the other, take placeinsidetheresistors.Thedropinpotentialintheconductingwiresthemselvesisusually negligible. Thus, for all intents and purposes, good conductors, and wires made out of good conductors,actasiftheyhavezeroresistance. SampleOhmsLawProblems Aninevoltbatterysuppliespowertoacordlesscurlingironwitharesistanceof 18ohms.Howmuchcurrentisflowingthroughthecurlingiron?0.5amps A110voltwalloutletsuppliespowertoastrobelightwitharesistanceof2200ohms. Howmuchcurrentisflowingthroughthestrobelight?0.05amps MFCscommonlyachieveaworkingvoltageof0.30.7V.Thevoltage(orcellpotential)is afunctionoftheexternalresistanceorloadonthecircuitandthecurrent.Ohmslawcan also be used to calculate the cell potential for the MFC. The current produced from a singleMFCisusuallysosmallthatwhenconstructedinalaboratory,thecurrentfromthe MFCisnotmeasured,butiscalculatedfromthemeasuredvoltagedropacrosstheresistor as I = V/R. The highest voltage produced in the MFC is called the open circuit voltage, OCV, which is measured with the circuit disconnected (infinite resistance, zero current). As the resistances are decreased, the voltage decreases. The power at any time is calculatedasP=IV,whichwillbedescribedinmoredetaillater(seeChapter1Powerin ElectricalCircuits). CIRCUITDIAGRAMS Electric circuits, whether simple or complex, can be described with mere words saying something like, "A light bulb is connected to a Dcell" is a sufficient amount of words to describeasimplecircuit.But,anothermeansofdescribingacircuitistosimplydrawit.Such drawings,calledcircuitdiagrams,provideaquickermentalpictureoftheactualcircuit. Describing Words Circuits with DescribingCircuitswithDrawings

"A circuit contains a light bulb anda1.5VoltDcell."

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A final means of describing an electric circuit is by use of conventional circuit symbols to provide a schematic diagram of the circuit and its components. Some circuit symbols used in schematicdiagramsareshownbelow.
SingleCellBatteriesConnectingWireResistorSwitch(open)Switch(closed)

Asinglecellorotherpowersource,aspreviouslynoted,isrepresentedbyalongandashort parallelline.Acollectionofcells,orbatteries,isrepresentedbyacollectionoflongandshort parallellines.Inbothcases,thelonglineisrepresentativeofthepositiveterminaloftheenergy sourceandtheshortlinerepresentsthenegativeterminal.Astraightlineisusedtorepresenta connectingwirebetweenanytwocomponentsofthecircuit.Anelectricaldevice,whichoffers resistancetotheflowofcharge(aresistor),isrepresentedbyazigzagline.Anopenswitchis generally represented by providing a break in a straight line by lifting a portion of the line upwardatadiagonal.Asanillustrationoftheuseofelectricalsymbolsinschematicdiagrams, considerthefollowingtwoexamples. Example1: Description using words: Three Dcell batteries are used to power a circuit containing three lightbulbs.
DrawingofCircuitSchematicDiagramofCircuit

Using the verbal description, one can acquire a mental picture of the circuit being described. Thisverbaldescriptioncanthenberepresentedbyadrawingofthreecellsandthreelightbulbs connectedbywires.Finally,thecircuitsymbolspresentedabovecanbeusedtorepresentthe samecircuit.Notethatthreesetsoflongandshortparallellineshavebeenusedtorepresent the three Dcells. Also note that each light bulb is represented by its own individual resistor symbol. Straight lines have been used to connect the two terminals of the battery to the resistorsandtheresistorstoeachother. Theabovecircuitspresumedthatthethreelightbulbswereconnectedinsuchawaythatthe charge flowing through the circuit would pass through each one of the three light bulbs in consecutivefashion.Thepathofapositivechargeleavingthepositiveterminalofthebattery and traversing the external circuit would involve a passage through each one of the three connectedlightbulbsbeforereturningtothenegativeterminalofthebattery.Butisthisthe onlywaythatthreelightbulbscanbeconnected?Dotheyhavetobeconnectedinconsecutive

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fashionasshownabove?Absolutelynot!Infact,example2belowcontainsthesameverbal descriptionwiththedrawingandtheschematicdiagramsbeingdrawndifferently. Example2: Description using words: Three Dcell batteries are used to power a circuit containing three lightbulbs.
DrawingofCircuitSchematicDiagramofCircuit

Again, using the verbal description, one can acquire a mental picture of the circuit being described, however this time, the connections of light bulbs are done in a manner such that there is a point on the circuit where the wires branch off from each other. The branching locationisreferredtoasanode.Eachlightbulbisplacedinitsownseparatebranch.These branchwireseventuallyconnecttoeachothertoformasecondnode.Asinglewireisusedto connectthissecondnodetothenegativeterminalofthebattery. Thesetwoexamplesillustratethetwocommontypesofconnectionsmadeinelectriccircuits. When two or more resistors are present in a circuit, they can be connected in series or in parallel. Activity1.2CreatingElectricalCircuits Students will sketch and construct several circuits and draw corresponding circuit diagramstoinvestigateparallelandseriescircuits.

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Activity1.2CreatingElectricalCircuits
ProblemA

TypeofCircuit:_____________________________

Sketch and construct an electric circuit in which one Dtype battery can light two bulbs asbrightlyasonebulb.

ProblemB

TypeofCircuit:_____________________________

Sketch and construct an electric circuit in

whichonebulbislitbrighterwithtwoDtype batteriesthanwithoneDtypebattery.

ProblemC

TypeofCircuit:_____________________________

Sketch and construct an electric circuit in

whichonebulbislightednobrighterwithtwo DtypebatteriesthanwithoneDtypebattery.

ProblemD

TypeofCircuit:_____________________________

Sketch and construct an electric circuit in

whichoneDtypebatterylightstwobulbsless brightlythanonebulb.

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Activity1.2CreatingElectricalCircuits(withanswers)
ProblemA

PARALLEL

Sketch and construct an electric circuit in Rationale:Twobulbswiredinparallelareequally which one Dtype battery can light two bulbs bright and have the same brightness as the standard.Thevoltageisevenlydistributedacross equallyasbrightasonebulb.
theentirecircuit.

ProblemB

SERIES

Sketch and construct an electric circuit in Rationale: Two batteries wired in series have whichonebulbislightedbrighterwithtwoD twice the voltage as a single cell. The voltage is additive. typebatteriesthanwithoneDtypebattery.

ProblemC

PARALLEL

Sketch and construct an electric circuit in Rationale: Two batteries wired in parallel have whichonebulbislightednobrighterwithtwo thesamevoltageasasinglecell,butlasttwiceas DtypebatteriesthanwithoneDtypebattery. long. The voltage is evenly distributed across the entirecircuit.

ProblemD

SERIES

Sketch and construct an electric circuit in Rationale:Twobulbswiredinseriesareequally whichoneDtypebatterylightstwobulbsless bright, but dimmer than the standard. The voltageisadditive. brightlythanonebulb.

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Activity1.2CreatingElectricalCircuits(InstructorNotes)

Have the students observe both current and voltage with a multimeter as they construct thecircuits. Askthemtomakeobservationsaboutthedifferencesbetweenparallelandseries connectionsastheyrelatetoOhmsLaw(takingintoaccounttheresistanceofthebulbs).

QuestionsforAnalysis Whatarethebasiccomponentsofanelectricalcircuit? Twoormoreinterconnectedcomponentsconsistingofasourceofcurrentor voltageandaresistor. Whatisthedifferencebetweenseriesandparallelcircuits? Aseriescircuitisasinglepathforelectriccurrentthroughallofitscomponents. Several batteries connected in series can produce greater voltage than a single battery can. A parallel circuit is a different path for current through each of its components. A parallel circuit provides the same voltage across all its components. YoumayhavenoticedthatanentirestringofoldChristmaslightsgoesoutifonebulb burnsoutorisremoved.Explain. Christmaslightsareconnectedinseries,sowhenonegoesoutorisremoved,an opencircuitiscreated.Anopencircuitisanincompletecircuitduetoabreakin thecontinuousconnectionofconductorsfromoneendofanelectricalsourceto theother.

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SERIESANDPARALLELCIRCUITS Aseriescircuitisacircuitinwhichresistorsarearrangedinachainsothatthecurrenthasonly one path to take. The current is the same through each resistor. The total resistance of the circuitisfoundbysimplyaddinguptheresistance(orvoltage)valuesoftheindividualresistors: equivalentresistanceofresistorsinseries:R=R1+R2+R3+... R1R2R3 V I Aseriescircuitisshowninthediagramabove.Thecurrentflowsthrougheachresistorinturn. Ifthevaluesofthethreeresistorsare: R1=8,R2=8,andR3=4,thenthetotalresistanceis8+8+4=20. Witha10Vbattery,byV=IR,thetotalcurrentinthecircuitis: I=V/R=10/20=0.5A.Thecurrentthrougheachresistorwouldbe0.5A. A parallel circuit is a circuit in which the resistors are arranged with their heads connected together, and their tails connected together. The current in a parallel circuit breaks up, with someflowingalongeachparallelbranch,andrecombiningwhenthebranchesmeetagain.The voltageacrosseachresistorinparallelisthesame. Thetotalresistanceofasetofresistorsinparallelisfoundbyaddingupthereciprocalsofthe resistancevalues,andthentakingthereciprocalofthetotal: equivalentresistanceofresistorsinparallel:1/R=1/R1+1/R2+1/R3+... I1R1 II2R2I I3R3 V

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Aparallelcircuitisshowninthediagramabove.Inthiscase,thecurrentsuppliedbythebattery splitsup,andtheamountgoingthrougheachresistordependsontheresistance.Ifthevalues ofthethreeresistorsare: R1=8,R2=8,andR3=4,thenthetotalresistanceisfoundby 1/R=1/8+1/8+1/4=1/2.ThisgivesR=2. Witha10Vbattery,byV=IR,thetotalcurrentinthecircuitis:I=V/R=10/2=5A. TheindividualcurrentscanalsobefoundusingI=V/R.Thevoltageacrosseachresistoris10V, andtherefore: I1=10/8=1.25A I2=10/8=1.25A I3=10/4=2.5A Notethatthecurrentsaddtogetherto5A,thetotalcurrent. Inamanneranalogoustoresistors,batteries(orsourcesofvoltage)canalsobeplacedwithin circuits in series and in parallel. In each instance, batteries added in series increase voltage, whilebatteriesaddedinparallelincreasecurrent.
ThereasonourexperimentalMFChastwoanodechambersisanalogoustotheconnection of two batteries in series more voltage isgenerated. To light the LED, approximately 0.45 V is necessary, and a single anode chamber often is not capable of supplying that muchvoltage.Thereasontheanodechambersaresituatedoneithersideofthecathode chamber is to provide increased surface area at the interface between the anode and cathodeforthemigrationofpositiveionsthroughtheionexchangemembrane.

EMFANDINTERNALRESISTANCE Real batteries are constructed from materials, which possess nonzero resistivities, so real batteriesarenotjustpurevoltagesources.Theyalsopossessinternalresistances.Incidentally, apurevoltagesourceisusuallyreferredtoasanemf(whichstandsforelectromotiveforce).Of course,emfisalsomeasuredinunitsofvolts.Abatterycanbemodeledasanemfconnected inserieswitharesistorr,whichrepresentsitsinternalresistance.Supposethatsuchabattery isusedtodriveacurrentIthroughanexternalloadresistorRasshowninthecircuitdiagram below. Note that in circuit diagrams, an emf is represented as two closely spaced parallel linesofunequallength.Aresistorisrepresentedasazigzagline.

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 Battery B

Considerthebatteryabove.ThevoltageVofthebatteryisdefinedasthedifferenceinelectric potentialbetweenitspositiveandnegativeterminals:i.e.,thepointsAandB,respectively.As wemovefromBtoA,theelectricpotentialincreasesby+ voltsaswecrosstheemf,butthen decreases by1/rvolts as we cross theinternal resistor. Thus,the voltage Vof thebatteryis relatedtoitsemfandinternalresistancervia: V = - IR Now, we usually think of the emf of a battery as being essentially constant (since it only dependsonthechemicalreactiongoingoninsidethebattery,whichconvertschemicalenergy into electrical energy). Therefore, we must conclude that the voltage of a battery actually decreasesasthecurrentdrawnfromitincreases.Infact,thevoltageonlyequalstheemfwhen thecurrentisnegligiblysmall. Ifweshortcircuitabattery,byconnectingitspositiveandnegativeterminalstogetherusinga conductingwireofnegligibleresistance,thecurrentdrawnfromthebatteryislimitedonlyby its internal resistance. In fact, in this case, the current is equal to the maximum possible current. A real battery is usually characterized in terms of its emf (i.e., its voltage at zero current)andthemaximumcurrentthatitcansupply.Forinstance,astandarddrycell(i.e.,the sort of battery used to power remote controls) is usually rated at 1.5 V and (around) 0.1 A. Thus,nothingreallycatastrophicisgoingtohappenifweshortcircuitadrycell.Wewillrun thebatterydowninacomparativelyshortperiodoftime,butnodangerouslylargecurrentis goingtoflow.Ontheotherhand,acarbatteryisusuallyratedat12Vandsomethinglike200A (thisisthesortofcurrentneededtooperatethestartermotor).Itisclearthatacarbattery musthaveamuchlowerinternalresistancethanadrycell.Itfollowsthatifwewerefoolish enoughtoshortcircuitacarbattery,theresultwouldbefairlycatastrophic(imagineallofthe

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energyneededtoturnovertheengineofacargoingintoathinwireconnectingthebattery terminalstogether). Problem: What is the terminal (final) voltage supplied by a cell of emf 2.0 V with an internalresistanceof1.0whenitisconnectedtoa9.0resistor? BeginbyusingOhmslawtofindthecurrentsuppliedbythebattery. I=V/R=2.0/9.0=0.22A Then,usetheinternalresistanceformulatosolvetherestoftheproblem. V = - IR V=2.0(0.22)(1)=1.8V It is not unusual to note significant differences in voltage output between differing MFC assemblies based on specific system architecture. The main reason is because of the internalresistanceofthereactorcomparedtothemaximumpossiblepotentialduetothe chemicalreactionsattheanodeandcathode,orthecellpotential,Eemf(seeChapter1 StandardReductionPotentials).Todeterminethetotalvoltagecapableofbeingproduced bythecell,wemustviewtheMFCashavingcurrentthroughtworesistorslinkedinseries, withonebeingtheexternalloadandtheothertheinternalresistance. BeingthattheoperationalconditionswithintheMFCaredefinedbytherequirementsof themicrobesforoptimalgrowthandmetabolismandarenotnecessarilycompatiblewith themostfavorableelectrochemicalconditions,akeychallengeforthoselaboratoriesand scientists working to improve MFC voltage (and resulting power) outputs is to identify possible sources of internal resistance within the specific MFC design and significantly reducetheseresistances. POWERINELECTRICALCIRCUITS Inadditiontovoltageandcurrent,anothermeasureofelectronactivityinacircuitispower.In electric circuits, power is a function of both voltage and current. Not surprisingly, this relationshipbearsastrikingresemblancetoOhmslaw:

P=IV
Inthisequation,powerPisexactlyequaltothecurrentImultipliedbythevoltageV.When usingthisformula,theunitofmeasurementforpoweristhewatt,abbreviatedwiththeletter "W." It must be understood that neither voltage nor current by them selves constitute power. Rather, power is the combination of both voltage and current in a circuit. Remember that voltage is the specific work (or potential energy) per unit charge, while current is the rate at

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which electric charges move through a conductor. Voltage is analogous to the work done in lifting a weight against the pull of gravity. Current is analogous to the speed at which that weight is lifted. Thus, the rule is the power in a circuit is the product of the voltage and the current. According to the formula, a circuit with high voltage and low current may be dissipating the sameamountofpowerasacircuitwithlowvoltageandhighcurrent.Neithertheamountof voltagealone,northeamountofcurrentalone,indicatestheamountofpowerinanelectric circuit. In an open circuit, a circuit contains no external resistors, voltage is present between the terminalsofthebattery,andthereiszerocurrent.Asaresult,thereiszeropowerdissipated, nomatterhowgreatthatvoltagemaybe.SinceP=IVandI=0andanythingmultipliedby zero is zero, the power dissipated in any open circuit must be zero. Likewise, if we were to have a short circuit constructed of a loop of conducting wire (essentially zero resistance), we couldhaveaconditionofcurrentintheloopwithzerovoltageand,likewise,nopowerwould bedissipated. Thepowerformuladoesnotjustapplytobatteries.Itcouldalsoapplytoaresistorexternalto thebattery.Sucharesistorisreferredtoasaloadresistor.Itcouldbeeitheranelectriclight, anelectricheatingelement,or,maybe,anelectricmotor.Thebasicpurposeofthecircuitisto transfer energy from the battery to the load, where it actually does something useful for us (e.g.,lightingalightbulborliftingaweight). The power transfer between a voltage source and an external load (or load resistor) is most efficientwhentheresistanceoftheloadmatchestheinternalresistanceofthevoltagesource. If the load resistance is too low, then most of the power output of the voltage source is dissipatedasheatinsidethesourceitself.Electricalenergyisconvertedintoheat(i.e.,random motion of the atoms that make up the voltage source) as the electrically accelerated free electrons inside the source collide with the atoms and, thereby, transfer all of their kinetic energy to the atoms. If the load resistance is too high, then the current, which flows in the circuit,istoolowtotransferenergytotheloadatanappreciablerate.Theoptimalcaseexists whentheloadresistanceisequaltotheinternalresistanceofthevoltagesource,however,only halfofthepowerofthevoltagesourceistransferredtotheload.Theotherhalfisdissipatedas heatinsidethesource.

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To make MFCs useful as a method to generate power, it is necessary to first optimize the systemforpowerproduction.PoweriscalculatedfromthevoltageandthecurrentasP=I E, where E stands for the cell potential (in some instances, V is not used for voltage becausethesymbolandtheunitsV=Voltscanleadtoconfusion).

The power generated by an MFC is calculated from the measured voltage, EMFC, across the load and the current using P = I EMFC. The current produced by a laboratoryscale MFC is commonlycalculated,aspreviousstated,bymeasuringthepotentialacrosstheload(i.e.,the externalresistor,Rext),andusingI=EMFC/Rext.Thus,poweroutputcanbefoundusing

BasedontherelationshipI=EMFC/Rext,wecanalternativelyexpresspoweroutputinterms ofthecalculatedcurrentas:

P=I2Rext

Activity1.3.A(Assessment)TheShoeboxRoom Activity1.3.B(Assessment)CanYouLightYourWay? Students will demonstrate an understanding of the requirements of making an electrical circuit;will be able to build a working series circuit and parallel circuit;will be able to describe how both types of circuits work;will be able to describe the advantages and disadvantages of parallel and series circuits;will understand that electrical energy can be convertedtoheat,light,soundandmotion;andwillwireashoeboxhousewithseriesand parallelcircuitsto operatelightsandotherresistorsthatarecontrolledbyswitches.

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Activity1.3.ATheShoeboxRoom
Hello,mynameisStuart.Iliveinaverysmallhouse,andyouwontfindmecomplainingabout that.Imsuremylittlehouseismuchmorecomfortablethanyouraveragecaveormudhut. Houses, as Im sure youve noticed, seem to be getting smaller and smaller and smaller until eventually,Iimagine,wewillallbelivinginshoeboxes.Notjustme. Yes,althoughtheentirehousehasnotyetshrunkintosomethingyoucouldeasilywalkaround with,mybedroomisridiculouslytiny.Itisbyfarthesmallestinthehouse. Iamlookingtodosomeredecoratingandremodelinginmybedroomandwouldliketoinclude afewnewamenitiesandupgradestoitscurrentdesign: Analarmthatsoundswhenthedooropens Aceilingfanthatcanrunonmorethanonespeedwithalight Areadinglampwithmorethanonelevelofbrightness

Beingthatmostinteriordecoratingbusinessesandelectricalengineeringfirmsdontdealwith rooms as small as shoeboxes, Stuart has sought you out to design and construct his new bedroom. You should begin by sketching out a floor plan for the room, including the location of all the furnitureandfixtures,thendrawingoutcircuitdiagramsforthenecessaryelectricalwork,and, finally,completingtheelectricalworkwithintheroom. InadditiontocreatingafullyfunctioningshoeboxroomforStuart,hehasalsoaskedthatyou prepare a report detailing how the electrical energy is being transferred and transformed throughout his room. The report should also include a discussion of wiring choices, resistor arrangements,andenergyflow. ThefollowingmaterialswillbeprovidedforyoutouseinthedesignofStuartsshoeboxroom: Fortheteachertodevelop: Needtodevelopmaterialslist Needtodevelopscoringrubric

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Activity1.3.BCanYouLightYourWay?
(afterProf.G.Watson,UniversityofDelaware,www.pysics.udel.edu)

"What'swrongwiththecarbidelantern?" BeforeChriscouldutterareplytoPat'squestion,darknesssuddenlydescended. "Great!I'vebeenbeggingmyfolksforanewlampsincethelasttimewewentcaving.Toolate now!Let'sfireuponeofourcandles." It took a bit of rummaging through the pack to find one in the shroud of darkness. Finally ready, Pat struck a match but it immediately flickered out. This happened again with the secondmatch,andagainwiththethird.ThereasonsuddenlyoccurredtoChris. "You've gotta be kidding me I think the air is bad down here! The guidebook didn't say anythingaboutthat,didit?" "Not that I remember. You know, my breath does seem a little short right now that you mentionit.Thinkwemadeawrongturn?" "Don'tknow,butweneedtogetoutofhererightnow.ShineyourflashlightoverherewhileI takeanotherlookatthemap." AminutepassedwhilePatrifledthroughthepackagain. "Badnews!Imusthavelefttheflashlightlyingonthegroundwhereweateourlunch." "You'dbetterbejoking!Wehavewalkedforfourhourssinceweate.Wecan'tmakeitback therewithoutalight." "I'mserious.Iremembertakingitouttogetthesandwiches.GuessIforgottoputitback." "Nevermind,Ikeepasmallflashlightinmyjacket.Hey,whereisit?Darnit,mylittlebrotheris alwaysstealingmystuff!" "Calmdown!Gettingupsetwonthelpanything!" "OK.OK.Youreright.Whatdowehaveforalight?" "Well, I do have a spare bulb for my lantern in my pocket. But I didn't bring a spare battery; they'resoheavy."

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"Do you have anything else with batteries in it? I've got two extra AAA batteries for the flashlightthatlittlethieftook." "Ivegotnothingelsewithbatteries.Idontcarryalittlebitofeverythinglikeyoudo." "GoodthingIdo!Butthebulbisfora6Vbattery.DoyouthinkitwillworkwithtwoAAAcells?" "Even if it does, do you think it will burn long enough for us to get out? I'm starting to get worried..." "Maybewecanuseoneatatimetomakethemlastlonger.Itwon'tbeasbrightandwellhave to walk slower, but maybe one at a time would be better. Hey, youve been learning about electricalcircuitinschool.Whatshouldwedo?" TheExercise Yourlabexercisecomesintwoparts. 1. YourgroupwillhaveavailabletwoAAAcellsandthebulbdescribedabove.Inthelab youwillhaveavailabletwomultimeters,lengthsofwire,andachoiceofresistors.Make themeasurementsnecessarytoanswerthequestionsintheprecedingstory.Youmay wishtoconsultadditionalonlineresourcesforsupportingdata. 2. Designaflashlightusingmaterialsthatyouwishyouhadinyourpockets.Becreative assumethatyouarenotcarryingalengthofwire!Also,aswitchingmechanismwould beadesirablefeature.Inthenextclassperiod,yourgroupwillbegivenfiveminutesto assembleaworkingflashlight.Andwemayeventurnoutthelights!!!

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ELECTROCHEMICALCELLS

Anelectrochemicalcellgeneratesanelectromotiveforceoremf(voltage)andelectriccurrent from chemical reactions. The current is caused by the chemical reactions releasing and accepting electrons at the different ends of a conductor. Electrochemistry is the branch of chemistryinvolvedwiththestudyofelectricityproducedbychemicalreactions. There are two different kinds of electrochemical cells. Electrolytic cells are those in which electrical energy causes nonspontaneous chemical reactions to occur, and voltaic cells are those in which spontaneous chemical reactions produce electricity and pass it through an external circuit. In a voltaic cell, electrical current can be conducted through metal wires or alongmetalsurfacesandthroughpureliquidelectrolytesorsolutionsthatcontainelectrolytes. When a metallic conductor is employed, current is conducted through the metal without causinganychemicalchange.Electrolyticconductionoccursviathemotionofionsthroughthe solutionorpureliquid. Thechemicalreactionsinbatteriesoccurwhentwodissimilarmaterials,suchasaluminumand copper (called electrodes) react together when inserted into a chemical conduction solution called an electrolyte. The electrolyte solution begins to slowly dissolve the aluminum electrodes,yieldingpositivealuminumionsandelectrons.Theseelectronsmustremainbound to an electron conducting material and cannot move into the electrolyte. The positive ions accumulateintheelectrolyteandflow(viaconcentrationgradient)towardsthecathode.The electrons travel around, from the anode, through the conducting wires, and complete the circuit at the cathode. At the cathode, the positive aluminum ions in combination with the electronscausealuminumtobedepositedonthecopperelectrode.
Activity1.4AHomemadeLemonyBattery Studentswillconstructafunctionalbatteryfromseveralcommonhouseholditems.

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Activity1.4AHomemadeLemonyBattery

Materials: lemons,coins(suchascopperpennies),papertowels,aluminumfoil(orcoins,suchasdimes), bowls, scissors, lemon juicer, wire strippers, plastic tape, paper tube (toilet paper or paper toweltubewillwork),plasticcoveredelectricalwire Procedure: 1.Wrapfoiloveroneendofthepapertubeandthensecureitbytapingitdown. 2.Withthewirecutters,strip12oftheplasticfromthewire.Tapeoneendtothefoil.Then, setupthepapertubewiththefoildownandtheopeningontop. 3.Squeezethejuicefromthelemonsintoadish.Soakthepapertowelsinthejuice.Then,start fillingthepapertubewithasmallrolledpieceoftoweling,thenacoin,followedbyapieceof foil.Continuetolayerthesethreematerials,fillingthetubeandendingwithacoin.Tapea secondstrippedwiretothecoin. 4.Moistenafingertiponeachhandandtouchtheendsofthetwowires.Studentswill experienceasmallshockortinglebutitwillbeveryharmless.Studentsmayalsotryattaching anLEDtoeachendofthebattery.

QuestionsforAnalysis

IfyouarenotgeneratingenoughelectricitytolighttheLED,whatcouldyoudoto generatemorevoltage? Connectseveralhomemadebatteriestogetherinseries.

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Thehomemadebatteryisanexampleofawetcell.Thelemonjuiceactsastheelectrolytethat conductstheelectricitycreatedbythecoinsandthefoil.Wetcellsarecharacterizedbyliquid electrolytes.

Whilelemonjuiceactedasthesoleelectrolyteinthishomemadebattery,ourMFCwill beusingtwodifferentelectrolytes,oneintheanode(negative)chamberandoneinthe cathode(positive)chamber.Theanodewillcontainamixtureofsoilbacteriainaglucose nutrient growth media with an added redox mediator (methylene blue). The cathode willcontainasolutionofasolublechemicalelectronacceptor(oxidizer)calledpotassium ferricyanide. The MFC must utilize two different electrolytes because of the nature of the reactions occurring in each chamber. In the anode chamber, there is no oxygen present (anaerobic conditions), while there typically is oxygen present in the cathode chamber. Some MFC designs can also have anaerobic cathode chambers, e.g. with chemicalelectronacceptors,suchasferricyanide.
Activity1.5TheHandBattery

Studentswillobservethatvoltageoutput(i.e.,batteries)canresultfrommanydifferent Activity1.4TheHandBattery materialsaslongastheprinciplesofelectronflowareobserved. Materials: multimeter* or voltmeter (for instructions on operations and settings consult www.extech.com/instrument/products/400_450/manuals/EX410_UM.pdf or your instruction manual), copper plate and aluminum plate (each about the size of your hand), two electrical lead wires with alligator clips, a piece of wood or other nonmetallic surface, plates of other metals,suchastinorzinc(optional) * The multimeter must be capable of measuring low range current. Industrial gauges cannot alwaysreadinmilliampranges.Multimeterssensitiveenoughforthisactivitycanbepurchased atelectronicsstores,suchasRadioShack. Procedure: 1. Mount both metal plates on a piece of wood or simply clamp them to a nonmetallic surface.(Ifyouprefer,youdon'tevenhavetomountthetwoplates.Youcanattachthe wires as described below and then simply hold one plate in each hand. This has the benefitofallowingyoutosubstituteothermetalseasily.)

Activity1.5TheHandBattery

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2. Usingtheclipleads,connectoneplatetooneofthemeter'sterminalsandconnectthe otherplatetotheotherterminal.Atthispoint,itdoesn'tmatterwhichplateattachesto whichterminal. 3. Placeonehandoneachplate.Youshouldnoticeareadingonthemeter.Ifthemeter doesn't show an electrical current, simply reverse your connections, attaching the copper plate to the terminal that the aluminum was connected to and vice versa. If there is still no current, checktheconnectionsandthewiring. If that doesn't produce current,trycleaningtheplateswithapencileraserorsteelwooltoremoveoxidation. 4. Experiment with different metals to find out what combination produces the most current(amps).Trypressingharderontheplates.Getyourhandswetandtryagain. 5. Haveonepersonputahandonthecopperplateandanotherpersonputahandonthe aluminumplate,andthenhavethemjointheirfreehands. 6. Also,experimentwithmeasuringtheresistance(ohms)ofwetvs.dryhands. 7. Vigorously exercise (30 push ups or 30 jumping jacks), and once again measure the resistance. QuestionsforAnalysis Isthereadifferencebetweendryandwethandsinthevoltageorcurrent?Whyorwhy not? Wethands(lessresistance)shouldyieldahighercurrentandlowervoltagedue toOhmslaw. Whendidyouobtainthehighestandthelowestresistance? Wet(lowresistance),Dry(higherresistance) Istheresistanceofyourskinafunctionofmoistureinyourskin? Yes Suggesthowthehandbatterycouldbeutilizedasasimpletypeofliedetector. Onecouldassumethatapersonwhoislyingmaybenervous,causingtheirhands toperspire. Discussion: When you touch the two metal plates, the thin film of sweat on your hands acts as the electrolyte, reacting with the copper plate and with the aluminum plate. In one of these reactions, your hand takes negatively charged electrons away from the copper plate, leaving positivechargesbehind.Intheotherreaction,yourhandgiveselectronstothealuminumplate, causingittobecomenegativelycharged. Thisdifferenceinchargebetweenthetwoplatescreatesaflowofelectricalcharge,orelectrical current.Sinceelectronscanmovefreelythroughmetals,theexcesselectronsonthealuminum plate flow through the meter on their way to the copper plate. (In metals, positive charges 28

cannot move.) In your body, both positive and negative ions move. Negative electrons move throughyourbodyfromthehandtouchingthecoppertothehandtouchingaluminum.Atthe sametime,positiveionsmoveintheoppositedirection.Aslongasthereactionscontinue,the chargeswillcontinuetoflowandthemeterwillshowasmallcurrent. Your body resists the flow of current. Most of this resistance is in your skin. By wetting your skin,youcandecreaseyourresistanceandincreasethecurrentthroughthemeter.Sincetwo peopleholdinghandshavemoreresistancethanoneperson,theflowofcurrentwillbeless. If you disconnect one of the wires to the multimeter, the aluminum becomes negatively charged.Electronspileuponthealuminumsidebecausetheycannotcrossthegapinthewire. Thecopperbecomespositivelychargedasyourhandremoveselectronsfromthemetal.These pilesofchargecreateavoltage,whichismeasuredwhenthemultimeterissettoreadvolts. Most batteries use two different metals and an electrolyte solution to create a potential differenceand,thus,avoltage.Whentheterminalsofthebatteryareconnectedwithawire, thisvoltageproducesacurrent. Youcanuseotherpairsofdifferentmetalsinacircuittoproduceacurrent.Thesuccessyou haveusingvariousmetalswilldependonametal'selectricpotential,thatis,itsabilitytogainor lose charges. Try various metals to see which produces the highest current reading. An electromotiveseriestable(foundinmostchemistrytextbooks)showstheelectricpotentialsof metals and allows you to predict which metals will work well in making a hand battery (see table,page33). Youcansometimesgetasmallcurrentevenbetweentwoplatesmadeofthesamemetal.Each plate has a slightly different coating of oxides, salts, and oils on its surface. These coatings create slight differences in the surfaces of the metals, and these differences can produce an electricalcurrent.

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OXIDATIONREDUCTIONREACTIONS Batteriesrelyonchemicalreactionstogeneratevoltageandcurrent.Thecurrentiscausedby the reactions releasing and accepting electrons at the different ends of a conductor. The reactionisalwaysanoxidationreduction(orredox)reactioninvolvingatransferofelectrons. Redox reactions can be broken down into two halfreactions: (1) oxidation at the anode as electronsaretransferredfromanelectrondonortotheelectrode,and(2)reductionatcathode as an electron acceptor gains electrons from the electrode. Thus, in a spontaneous redox reaction, electrons flow from the oxidized reactant (electron donor) to the reduced reactant (electronacceptor).Theelectrodesaresimplytheordinarymetallicsurfacesuponwhichthe oxidationorreductionhalfreactionsoccur. Redoxprocessesoccurinsingledisplacementorsubstitutionreactions.Theredoxcomponent ofthistypeofreactionisthechangeofoxidationstate(charge)oncertainatoms,nottheactual exchangeofatomsinthecompounds. Inelectrochemicalcells,spontaneousoxidationreductionreactionsgenerateelectricalenergy. Forexample,inthereactionbetweenironandcopper(II)sulfatesolution: Fe + CuSO4 FeSO4 + Cu Theionicequationforthisreactionis: Fe + Cu2+ Fe2+ + Cu Astwohalfequations,itisseenthattheironisoxidized(electrondonor): Fe Fe2+ + 2e Andthecopperisreduced(electronacceptor): Cu2+ + 2e Cu Ifwedipanironstripintoacopper(II)sulfatesolution,theironwillbeeatenaway,aspongy layer of metallic copper will plate out on the iron strip, and the deep blue color of copper(II) sulfate will gradually fade. [Iron(II) sulfate, which is formed, is colorless.] In contrast, if we immerseacopperstripinaniron(II)sulfatesolution,noreactionwilloccurbecausethereverse reactionishighlynonspontaneous. Inthisarrangement,thespontaneoustransferofelectronsfromirontocopperisnotusefulfor generatingelectricalenergybecausetheenergyreleasedisdissipatedasheat.Itisanalogous toburningaspoonfulofsugarwithamatchinsteadofeatingitandconvertingthefreeenergy ofoxidationintousefulmusclework(seeChapter2MetabolismasaRedoxProcess).Ifsome means could be found to separate the removal of electrons from iron (oxidation) from the

30

donation of electrons to copper ions (reduction), then the electrons may be made to do somethingusefulalongtheway. Whenthetwohalfreactionsareseparated,thisflowofelectrons,insteadofoccurringatthe surfaceofthemetal,occursthroughanexternalcircuitandelectriccurrentisgenerated.Thisis calledavoltaiccell,andisexactlyhowabatteryworks.Thetwohalvesoftheredoxreaction arereferredtoashalfcells. A battery, like the ones found in a flashlight or calculator, contain oxidizing and reducing substances. As the electrons are transferred from anode to cathode, they are tapped to providethevoltagenecessarytopowertheelectricaldevice. VOLTAICCELLS Voltaic cells consist of two halfcells connected by a salt bridge. Each half cell is typically a metalelectrodeina1.0molar(1mol/LorM)aqueoussolutionofthemetalssalt.Thehalfcells areconnectedbyametalwireandavoltmetercanbeplacedintothecircuittomeasurethe potentialdifferencebetweenthetwoelectrodes.Thesaltbridgeisoftenemployedtoprovide electrical contact between two halfcells with very different electrolytes to prevent the solutionsfrommixingandismadeofsomemediumthroughwhichionscanslowlypassand maintain charge balance in the two halfcell solutions. A simple salt bridge consists of filter papersoakedwitharelativelyinertelectrolyte,usuallypotassiumchlorideorsodiumchloride. Another type of salt bridge consists of Ushaped glass tubes filled potassium chloride or sodiumchloride.Agarisoftenusedforgelificationwithinthetube. The two electrodes, connecting wire, and salt bridge form a closed circuit, with electrons moving from negative electrode to positive electrode through the wire, and positive and negativeionsmovingthroughthesaltbridge.Thus,electronswillflowspontaneouslyfromthe anode, where oxidation is occurring, to the cathode, where the reduction is occurring. Electricity in the voltaic cell is generated due to electric potential difference between two electrodes. The potential difference between the electrodes provides the driving force that pushestheelectronsthroughtheexternalcircuit.Thispotential,orEMF(electromotiveforce), of a cell is a measure of the tendency of the electrons to flow, and is dependant on the differenceinoxidizingorreducingstrengthofthetwohalfcells. A good analogy for the flow of electrons is the flow of water. Water flows spontaneouslydownhill.Damsandwaterwheelsareexamplesofwaysthatthe energy of flowing water is tapped to generate power. Cell potential is roughly analogous to a height difference between two reservoirs of water that are connected. Water will have a tendency (potential) to flow between the reservoirs,andthatflowcanbetappedtodowork. To maintain electrical neutrality and complete the circuit in the ironcopper cell described above, two Cl ions from the salt bridge migrate into the anode solution for every Fe2+ ion

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formed. Simultaneously, two K+ ions migrate into the cathode solution to replace every Cu2+ ionreduced.
IntheMFC,electronsaregeneratedattheanodeasglucoseisoxidizedthroughaprocess calledglycolysis(seeChapter2Step1Glycolysis).Theelectronsarethenshuttledto the anode with the help of a chemical redox mediator called methylene blue. Copper wires, connected to the electrodes by alligator clips, conduct the electrons, potentially througharesistor(LED),tothecathode.Inthecathodechamber,electronsaredischarged from the cathode to potassium ferricyanide, or, more specifically, the ferricyanide ion, Fe(CN)63, is reduced to the ferrous cyanide ion, Fe(CN)64. In our MFC, the circuit is closed, not by a salt bridge, but rather by semiporous, ionexchange membranes that separatetheanodeandcathodechambers.Ifthepotentialdifferencebetweentheanode andcathodechambersislargeenough(over0.45Vrequired),theMFCcanpowertheLED.

Voltaiccellscanberepresentedinshorthandform: Fe|Fe2+(1.0M)||Cu2+(1.0M)|Cu Theanode(Fe)isshownontheleftsideandthecathode(Cu)ontherightside.Theelectrode surfacesandthespecies(andtheirconcentrations)incontactwiththeelectrodesurfacesare separatedbysingleverticallines.Doubleverticallinesrepresentthesaltbridge. STANDARDREDUCTIONPOTENTIALS ThereiscompetitionbetweenFe2+andCu2+intheironcoppervoltaiccellforelectrons.Itturns outthatCu2+iseasiertoreducethanFe2+ andwinsthebattlefortheelectrons.Thisisknown becauseeachhalfcellhasacharacteristicvoltageorpotential. Itwouldbedesirabletoseparatetheindividualcontributionseachhalfcellreactionmakesto the total cell potential. This would allow us to determine the relative tendencies of the particular oxidation or reduction halfreactions to occur. However, it is not possible to determine experimentally the potential of a single electrode since every oxidation must be accompanied by a reduction; that is, the electrons must have somewhere to go. As a consequence,itisnecessarytoestablishsomearbitrarystandard. Byinternationalagreement,thereferenceelectrodewasselectedtobethestandardhydrogen electrode(SHE).Thisstandardhydrogenelectrodeconsistsofapieceofmetalelectrolytically coated with a grainy, black surface of inert platinum metal immersed in a 1.0 M H+ solution. Hydrogen gas, H2, is then bubbled over the electrode at one atmosphere (atm) pressure. By convention,theSHEisassignedapotentialof0.00V,andthenallotherhalfcellpotentialsare measuredrelativetothatassignedvalue.

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H2(g)2H+(aq)+2e 2H+(aq)+2eH2(g)

Eo=0.00V Eo=0.00V

(SHEasanode) (SHEascathode)

TheCopperSHECell To determine the halfcell potential for the reduction of copper, a voltaic cell is constructed using a copper electrode and the standard hydrogen electrode. As the halfcell reactions ensue,thecopperelectrodedecreasesinmassandtheconcentrationofCu2+ionsdecreasesin the solution around the copper electrode. Simultaneously, gaseous hydrogen decreases in mass and the H+ ion concentration increases in the solution of the SHE. The total electrical potential for this cell has been experimentally determined to be +0.34 V at the start of the reaction. (anode) H22H++2e (cathode) Cu2+2e+Cu (cellreaction)H2+Cu2+2H++Cu Eo=0.00V Eo=? Eo=0.34V

Thus,thehalfcellpotentialforthereductionofcopperis Cu2++2eCuEo=+0.34V TheIronCopperCell As described earlier, in this cell, copper deposits on one electrode as the iron electrode decreasesinmass.Thetotalelectricalpotentialofthiscellhasbeencalculatedtobe+0.78V. FeFe2++2e Cu2++2eCu Fe+Cu2+Fe2++Cu Eo=? Eo=+0.34V Eo=+0.78V

Thus,thehalfcellpotentialfortheoxidationofironis FeFe2++2eEo=+0.44V Thereactionforthehalfcellisalwayswrittenasareduction(gainofelectrons) Zn2++2eZnEo=0.76V

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By measuring the potentials of other standard electrodes versus the SHE or some other standardhalfcellwhoseelectrodepotentialisknown,aseriesofstandardelectrodepotentials canbeestablished.Whentheelectrodesinvolvemetalsornonmetalsthatareincontactwith their ions, the resulting series is called the electromotive series or activity series of the elements(seethetablebelow).Thosespeciesatthetopoftheseriesandontherightsideof the reduction halfreaction are the most active and possess the greatest tendency to be oxidizedorloseelectrons(thebestreducingagents).Thespeciesatthebottomoftheserieson the left side of the equation undergo reduction easily to gain electrons (the best oxidizing agents). Any species on the left side of a given halfreaction will react spontaneously with a substance that is on the right side in halfreaction above it, and the voltaic cell will have a positivepotential.

StandardElectrodePotentialsat25oC TheElectromotiveSeries ElectrodeReaction Eo Li+(aq)+eLi(s) K+(aq)+eK(s) Na+(aq)+eNa(s) Mg2+(aq)+2eMg(s) Al3+(aq)+3eAl(s) Zn2+(aq)+2eZn(s) Cr3+(aq)+3eCr(s) Fe2+(aq)+2eFe(s) Cd2+(aq)+2eCd(s) Ni2+(aq)+2eNi(s) Sn2+(aq)+2eSn(s) Pb2+(aq)+2ePb(s) 2H+(aq)+2eH2(s) Cu2+(aq)+2eCu(s) I2(s)+2e2I(aq) Fe3+(aq)+e Fe2+(aq) Hg2+(aq)+2eHg(l) Ag+(aq)+eAg(s) Br2(l)+2e2Br(aq) Cl2(g)+2e2Cl(aq) Au3+(aq)+3eAu(s) F2(g)+2e2F(aq) 3.05V 2.93V 2.71V 2.38V 1.66V 0.76V 0.74V 0.44V 0.40V 0.25V 0.14V 0.13V 0.00V +0.34V +0.54V +0.77V +0.79V +0.80V +1.07V +1.36V +1.50V +2.87V

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Problem:Whatisthespontaneouselectrochemicalreactionthatoccurswhenastandard copperhalfcelliscombinedwithastandardsilverhalfcellandwhatisthevalueofEoforthis voltaiccell? Fromthetableweknowthat Cu2++2eCuEo=+0.337V Ag++eAgEo=+0.800V Combiningbothhalfcells,wehavethefollowingequationtogivetheanswer CuCu2++2eEo=0.337V 2Ag++2e2AgEo=+0.800V Cu+2Ag+Cu2++2AgEo=+0.463V Notethatdoublingthesilverhalfcellreactionbalancestheelectrons,butdoesnotaffect thevalueofEoforthehalfreaction. NONSTANDARDSTATEREDUCTIONPOTENTIALS TheNernstequationwasderivedtoobtainelectrodepotentialsforconcentrationsandpartial pressuresotherthanthestandardstatevalues. Ecell=Eo2.303(RT/nF)logQ Ecell=theelectrodepotentialundernonstandardconditions Eo=thestandardelectrodepotential R=thegasconstant,8.314J/moleK T=theabsolutetemperature,inK n=thenumberofmolesofelectronstransferredpermoleofcellreaction F=theFaradayconstant,96485J/Vmolee Q=thereactionquotient(concentrationofproductsdividedbyconcentrationof reagents) PuttingthevaluesgivenaboveintotheNernstequationat298K(25oC)simplifiestheequation toEcell=Eo(0.0592/n)logQ. Problem:CalculatethereductionpotentialfortheFe3+/Fe2+electrodeiftheconcentration ofFe2+isfivetimeslargerthanthatofFe3+. Fromatableofstandardreductionpotentials,weknowthat Fe3++eFe2+Eo=+0.771V UsingtheNernstequation,wecansolvethealgorithmtofindtheanswer Ecell=Eo(0.0592/n)logQ Ecell=+0.771V(0.0592/1)log5=+0.771V0.0592(0.699) Ecell=+0.771V0.041V=+0.730V

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 Problem: A voltaic cell is prepared by combining the Fe3+/Fe2+ couple with the MnO4 /Mn2+ couple. In one compartment of the cell, [Fe3+] = 1.00 M and [Fe2+] = 0.100 M. In the othercompartment,[MnO4]=1.00x102M,[Mn2+]=1.00x104,and[H+]=1.00x103M. ThecellreactionisMnO4+8H++5Fe2+Mn2++4H2O+5Fe3+,withEo=+0.74V.Whatisthe cellpotential,E,forthecell? Ecell=Eo(0.0592/n)log[Mn2+][Fe3+]5/[MnO4][H+]8[Fe2+]5 Ecell=+0.74(0.0592/5)log(1.00x104)(1.00)5/(1.00x102)(1.00x103)8(1.00x101)5 Ecell=+0.74(0.0592/5)log(1.00x1027)=+0.74V0.32V Ecell=+0.42V Problem:ThenickelleadvoltaiccellundergoestheelectrochemicalreactionNi+Pb2+ Ni2+ + Pb with Eo = + 0.124 V. If the concentration of Pb2+ in the cell is 1.00 M and the cell potential,E,is0.183V,whatistheconcentrationofNi2+inthecell? Ecell=Eo(0.0592/n)log[Ni2+]/[Pb2+] 0.183=0.124(0.0592/2)log[Ni2+]/(1.00) 0.059=0.0296log[Ni2+]/(1.00) 0.059/(0.0296)=2.0=log[Ni2+]/(1.00) [Ni2+]/(1.00)=1.0x102 [Ni2+]=1.0x102M ThevoltagegeneratedbyanMFC(andtheresultingpoweroutput)isfarmorecomplicatedto predict or calculate than that of a standard voltaic cell or even a chemical fuel cell. In an MFC, the bacteria colonizing the anode chamber must grow and manufacture enzymes or structures capable of transferring electrons outside the cell (see Chapter 3 Mediatorless MFCs). In a mixed culture, different bacteria can grow at different rates, setting different potentials.Additionally,themaximumvoltagesthatcanbegeneratedbytheMFCarebased on the specific electrochemical relationships between the electron donors (substrates) and acceptors(oxidizers). As noted previously, the open circuit voltage, OCV, measured for an MFC represents the maximumvoltagethatcanbeobtainedwiththesystem.TheOCVissubjecttothelimitations imposed by the specific bacterial community and the inherent electrochemical restrictions defined by the electron donor and acceptors. For an MFC, as with any power source, the objective is to maximize power output and, therefore, obtain the highest current possible under the conditions of maximum voltage. The OCV is only achieved under a condition of infiniteresistance.Whentheresistanceisreduced,thevoltagedropsproportionately.Thus, to maximize the power production, the key is to determine the smallest possible drop in voltageasthecurrentisincreasedoveraspecificrangeofcurrentinterest. To achieve this, a polarization curve is generated to characterize current as a function of voltage (Figure 1.4). By changing the circuit external resistance (load), a new voltage is obtained, and consequently, a new current at that resistance. Therefore, to obtain a polarization curve, a series of different resistance on the circuit is used and the voltage 36

measured at each resistance. The current is calculated using I = E / Rext and the voltage versusthecurrentplottedtogeneratethecurve.Thepolarizationcurveindicateshowwell theMFCmaintainsitsvoltageasafunctionofthecurrentproduction. Itshouldbenotedthatapolarizationcurveisveryeasilygeneratedusingsimpleresistorsthat can be purchased at a retail electronics supply store, such as Radio Shack, and then connectingtheresistorstothecircuitoftheMFCassembly. ApowerdensitycurveisthencalculatedfromthemeasuredvoltageasP=E2MFC/Rextor, alternatively, as P = I2 Rext. MFC researchers typically use the top of the power curve to reportthemaximumpowercapableofbeinggeneratedbyanMFC.

Figure 1.4.(A)Thecellpotential(voltage)wasplottedversuscurrent density to give the polarization curve, and then multiplied with each other(B)toobtainthepowerdensitycurve(P=E*I).Maximumpower isindicatedas1.0mWcm2.

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Activity1.6DemonstrationofaVoltaicCell:

Option1.6.A(simulation)Afterviewingavoltaiccellanimation,studentswillusea flashapplettosimulatevariousmetal/electrolytecombinationstocreateavoltaiccell withthegreatestcellpotential. Option1.6.B(wetlab)Studentswillconstructandevaluatethepotentialofseveral voltaic cells and compare their values to the accepted values found in a chemistry textbook.

Activity1.6.ATheVirtualVoltaicCell

Materials: copiesofstudentworksheet(downloadablefromsite),onePCperpairofstudents Procedure: 1. Ifyouhavethecapability,thetutorialcanbeshowntotheentireclassandtimegiven forclassdiscussion. 2. Directstudentstothefollowingwebsite: www.blackgold.ab.ca/ict/Division4/Science/Div.%204/Voltaic%20Cells/Voltaic.htm 3. Uponcompletionofthesimulation,studentsshouldcompletethestudentworksheet includedinthewebsite.

Activity1.6.BVoltaicCellsLab

Introduction An electrochemical reaction is a chemical reaction that involves reduction and oxidation (a redox reaction). The energy released in a spontaneous redox reaction can be used to do electrical work. This is accomplished using a voltaic (or galvanic) cell, a device in which the transfer of electrons occurs through an external pathway rather than directly between reactants.Aspontaneousredoxreactiontakesplacewhenastripofzincisputintoasolution containingcoppersulfate.ThereactionisZn(s)+Cu2+(aq)Zn2+(aq)+Cu(s).Carryingoutthe reactioninthisway,however,willnotpermittheobtainingofusefulelectricalenergy.Instead, azincstripisplacedintoasolutionofzincsulfateinonecontainer.Acopperstripisplacedinto a solution of copper sulfate in another container. An electrical conducting wire is connected

38

between the zinc and copper strips. A salt bridge containing an electrolyte capable of maintainingchargebalanceineachsolutionisplacedbetweenthetwosolutions.Avoltmeter canbeplacedinthecircuittomeasurethepotentialgeneratedinthereaction.Zincisoxidized toZn2+inonecompartment.Thezincstripiscalledtheanode.Cu2+isreducedtocopperinthe othercompartment.Thecopperstripiscalledthecathode.Theelectronslostbythezincto formZn2+travelfromtheanodethroughtheexternalcircuittothecathodeandarepickedup byCu2+ionsastheyformcopperatomsontheelectrode.Adiagramofthisvoltaiccellisshown inthefigurebelow: Theelectrochemicalreactioncanberepresentedastwohalfreactions: Zn(s)Zn2+(aq)+2e anode Cu2+(aq)+2eCu(s) cathode

Zn(s)+Cu2+(aq)Zn2+(aq)+Cu(s)overallreaction Ifallthecomponentsofthecellareintheirstandardstates(with[Zn2+]=1.0Mand[Cu2+]=1.0 M),thecelliscalledastandardcell.Thevoltagegeneratedbythestandardcelliscalledthe standardcellpotential(E). Itisconvenienttohaveashorthandmechanismfordesignatingvoltaiccells.Thestandardzinc coppercellcanbewrittenas: Zn(s)|Zn2+(1.0M)||Cu2+(1.0M)|Cu(s) In this designation, the anode, or oxidation halfcell, is represented on the left side and the cathode,orreductionhalfcell,isrepresentedontherightside.Theelectrodesinthetwohalf reactionsareelectricallyconnectedusingasaltbridgerepresentedbytwoverticallines.The 39

cellelectrodesarerepresentedatthetwoendsinthisnotation.Asingleverticallineshowsthe phaseboundarybetweenthesolidelectrodeandtheelectrolytesolutionineachhalfcell. PreLabProblem:WhatisthecellpotentialinaZn/Cucellthathasa[Zn2+]of0.10MintheZn halfcellanda[Cu2+]of0.010MintheCuhalfcell? Materials: Filterpaperdisks Dropperbottles Voltmeter Stripsofthefollowingmetals:copper,lead,silver,andzinc 1.0 Msolutionsofthefollowingsalts:CuSO4,Pb(NO3)2,AgNO3,ZnSO4,NaNO3 0.010Msolutionsofthefollowingsalts:CuSO4,Pb(NO3)2,AgNO3 I. StandardCells Six standard voltaic cells will be set up and the potential generated by each will be measured.However,insteadofsettingupthehalfcellsinbeakers,theywillbesetup onapieceoffilterpaper.Thehalfcellsareestablishedbyplacingthreedropsof1.0M solutionscontainingtherequiredmetalionsatfourspotsaroundthefilterpaper,and thensettingapieceoftheappropriatemetalincontactwiththesolution.Twodropsof 1.0MNaNO3areplacedonthecenterofthefilterpapertoserveasasaltbridge. Procedure: Obtainadiskoffilterpaper.Placethreedropsof1.0MCuSO4solutionatthetopofthe filter paper (at 12 oclock). Put a strip of copper in contact with the CuSO4 solution. Placethreedropsof1.0MPb(NO3)2solutionontherightsideofthefilterpaper(at3 oclock).PutapieceofleadincontactwiththePb(NO3)2solution.Transferthreedrops of1.0MAgNO3solutiontothebottomofthefilterpaper(at6oclock).Putapieceof silverincontactwiththeAgNO3solution.Transferthreedropsof1.0MZnSO4solution totheleftsideofthefilterpaper(at9oclock).PutazincstripincontactwiththeZnSO4 solution.Puttwodropsof1.0MNaNO3inthecenterofthefilterpapertoserveasthe saltbridge.Thearrangementwillbeasrepresentedinthefollowingfigure:

40

Turnthevoltmeterontothe2voltsDCrange.Placetheleadsofthevoltmetersuccessivelyon theCuandPbstrips,CuandAgstrips,CuandZnstrips,PbandAgstrips,PbandZnstrips,and, finally,AgandZnstripstomeasurethepotentialsoftheCu/Pb,Cu/Ag,Cu/Zn,Pb/Ag,Pb/Zn, andAg/Znstandardvoltaiccells,respectively.Ifyougetanegativepotential,switchtheleads onthemetalssothatyougetapositivepotential.Themetalthatyouconnecttotheblacklead ofthevoltmetertogetapositivereadingistheanodeofthecell.Recordthepotentialsfor eachofthecellsonthedatasheet,andcomparethemtotheliteraturevalues. II. NonstandardCells Ifoneormoreofthecomponentsofavoltaiccellisnotinitsstandardstate,thecellis notastandardcell.Thepotentialthatisexpectedtobegeneratedinsuchacellcanbe calculatedfromtheNernstequation,E=E0.0592logQ. Inthisequation,Eisthestandardcellpotential,Eisthecellpotential,nisthenumber ofelectronstransferredinthereaction,andQisthereactionquotient. Procedure: Obtainafilterpaperdiskandputthreedropsof1.0MAgNO3atthetop(at12oclock). PutapieceofsilverincontactwiththeAgNO3solution.Transferthreedropsof0.010M CuSO4 to the right side of the filter paper (at 3 oclock). Set a strip of copper on the CuSO4 solution. Place three drops of 0.010 M Pb(NO3)2 solution at the bottom of the filterpaper(at6oclock).PutapieceofleadincontactwiththePb(NO3)2solution.Put twodropsof1.0MNaNO3solutioninthecenterofthefilterpaper.Presstheleadsof thevoltmeterontothemetalstripsandmeasurethepotentialsoftheAg/CuandAg/Pb nonstandard voltaic cells. Record the values of the potentials on the data sheet. CalculatevaluesforthepotentialsexpectedusingtheNernstequation. ConcentrationCell Aconcentrationcellisavoltaiccellinwhichtheanodeandcathodecompartmentsare thesameexceptthattheconcentrationsofspeciespresentaredifferent.Forexample, a silver concentration cell contains a silver electrode in contact with a silver nitrate solution of a certain concentration in one compartment, and a silver electrode in contact with a silver nitrate solution of a different concentration in the other compartment.Thehalfcellpotentialgeneratedineachofthehalfcellsisgivenbythe Nernstequation,E=EAg0.0592log[Ag+]. n

III.

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Thepotentialoftheconcentrationcellisthedifferencebetweenthetwohalfcell potentials, Ecell=EAg(0.0592/1)log[Ag+]concentrated(EAg(0.0592/1)log[Ag+]diluted), or Ecell=0.0592log[Ag+]concentrated/[Ag+]diluted Procedure: Useadiskoffilterpaper,andputthreedropsof1.0MAgNO3onit.Placeapieceof silverincontactwiththe1.0MAgNO3solution.Transferthreedropsof0.010MAgNO3 to a different spot on the filter paper. Put a piece of silver on the 0.010 M AgNO3 solution.Addtwodropsof1.0MNaNO3betweentheothertwosolutions.Pressthe leads of the voltmeter against the two silver strips, and measure the potential of the cell.Recordthevalueofthepotentialonthedatasheet.Calculatetheexpectedvalues ofthepotentialfromtheNernstequation.

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Activity1.6.BStudentDataSheet
I. StandardCells VoltaicCell Pb(s)|Pb2+(1.0M)||Cu2+(1.0M)|Cu(s) Cu(s)|Cu2+(1.0M)||Ag+(1.0M)|Ag(s) Zn(s)|Zn2+(1.0M)||Cu2+(1.0M)|Cu(s) Pb(s)|Pb2+(1.0M)||Ag+(1.0M)|Ag(s) Zn(s)|Zn2+(1.0M)||Pb2+(1.0M)|Pb(s) Zn(s)|Zn2+(1.0M)||Ag+(1.0M)|Ag(s)

Experimental Literature Potential(Volts) Value(Volts)

II.

NonstandardCells

VoltaicCell Cu(s)|Cu2+(0.010M)||Ag+(1.0M)|Ag(s) Pb(s)|Pb2+(0.010M)||Ag+(1.0M)Ag(s) III.ConcentrationCell VoltaicCell Ag(s)|Ag+(1.0M)||Ag+(0.010M)|Ag(s)

Experimental Literature Potential(Volts) Value(Volts)

Experimental Literature Potential(Volts) Value(Volts)

PostLabQuestions: 1. Whywouldyouexpectnopotentialtobegeneratedbetweentwohalfcellsinthis experimentuntiltheNaNO3solutionmigratestoapointwhereitoverlapsbothof thehalfcellsolutions? 2. Discusssomepossiblesourcesoferrorthatcouldaccountforthedifferencesinthe experimentalvaluesofthecellpotentialsyouobtainedandtheliteraturevalues.

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Chapter 2: How do organisms obtain energy?orWhydontIrunonbatteries?

METABOLISMASAREDOXPROCESS Burningafuelisanoxidativeprocessandtheformationofafuelisareductiveprocess.These processesareperpetuallylinked.Thereisalwaysadonorandanacceptor.Asonesubstance becomes oxidized, the other substance is reduced and vice versa. The common feature between the two processes is the transfer of electrons. (In biology classes, the transfer of electrons, as it occurs in living cells, is almost always referred to as electron transport.) The substance that is being reduced accepts electrons donated by the substance that is being oxidized.Althoughtheactofgaininganelectron,whichiscalledareduction,maybeconfusing at first, the terminology starts to make more sense when we remember that electrons are negatively charged. Reduction implies becoming less and, obviously, accepting something negativecanbeinterpretedasadecrease. Respiration, whether carried out by humans or plants, is essentially the same process of burning or oxidizing organic compounds. The carbon containing compounds burned in respirationaremostlikelytobesugars.Thecombustionismorecontrolled(byenzymesthat keepthereactionsgoingatnondestructivetemperatures),butenergyisproducedandcarbon dioxideisliberatedjustasitwouldbeifsugarwereburnedonafire. Somebiologicaloxidationsaresimilartotraditionalcombustionreactionsinthattheyinvolve the direct addition of oxygen, but more often in living organisms, oxidation occurs by the removalofhydrogens.Becausehydrogeniscomposedofaproton(i.e.,ahydrogenion,H+)and anelectron,e,thetransferofhydrogensalwaysinvolvesthetransferofelectrons.

(atom)(proton)(electron)

HH++e

Inmetabolism,thereareoftenlongsequences,orchains,ofhydrogen(orelectron)acceptors. Thisisthereasonwhyburningsugarinrespiration,althoughiteventuallyinvolvestheaddition ofoxygen,doesnotliberateenergyquiteasdramaticallyasitdoesifsugarisburnedonafire. Instead,energyisreleasedinatrickleratherthanaflood.Inaddition,muchoftheenergyis not released at all, immediately, but is conserved, as chemical energy, in highenergy substances,suchasadenosinetriphosphateorATP. The characteristics of ATP make it exceptionally useful as the basic energy source of all cells. The three phosphate groups of ATP are the key to its ability to store and release energy. Wheneverthechemicalbondbetweenthesecondandthirdphosphategroupisbroken,energy isreleased,fuelingavarietyofcellularactivities.Theresultingadenosinediphosphate(ADP)is 44

identicaltoATPexceptithastwophosphategroupsinsteadofthree.Whenthepathwaysof cellularmetabolismcreatethosesmallamountsofenergy,alittleatatime,itcanbestoredby addingaphosphategrouptoADPmolecules,producingATP. Activity2.1(Demonstration)FruitSnackTerminator Studentswillobservetheactionofastrongoxidizer,KClO3,onsugarsandgainavery visualappreciationfortheamountofenergythatcanbeliberatedfromsugars.

Activity2.1FruitSnackTerminator
MATERIALS Onesmallpackageoffruitsnacks(anyvarietywillwork),ringstandandclamp,onemedium sizedpyrextesttube,around2.5gofsolidpotassiumchlorate(KClO3),safetyshieldand goggles,Bunsenburner&striker,tongsorlongforceps PROCEDURE Fillthetesttubetoadepthofaboutoneinchwithpotassiumchlorate.Clampthetesttubein place at an approximately 45 angle, directed away from any person. Set up the ring stand clamptesttubeassemblybehindasafetyshieldinfrontoftheclass.Connecttheburnersothat thetesttubecanbeheatedeasily. Lighttheburnerandheatthetesttubeatthebottomuntilthesolidmelts(m.p.isaround350 C). Stand behind the safety shield and carefully drop one fruit snack into the test tube using tongsorforceps.Aviolentflameshootingreactionensuesandlastsforaboutoneminute. DISCUSSION The fruit snack is mostly sugar, which is easily oxidized by something like molten potassium chlorate. Ideally, a balanced equation would show sucrose (C12H22O11) being converted to carbondioxideandwaterwhiletheKClO3becomesKCl.Theactualreactiondoesnotseemto go to total completion since there is usually a little gunky residue left behind from gelatin or agaradditives.

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HAZARDS MoltenKClO3cancauseverysevereburns.Exercisegoodsafetytechniquewhilepresentingthis demonstration.Thereisalsoalotofsmokeproducedduringtheoxidation,sothisexperiment shouldonlybedoneinaroomwithgoodventilation(becarefulinroomswithsmoke detectors). THEROLEOFGLUCOSEINMETABOLISM We, and all other living creatures, require a continuous source of chemical energy to remain alive. This is the reason for eating: We take in highly ordered molecules that have high free energy,andejectdisorderedmoleculeswithlowfreeenergy. All nutrition is based on one molecule, glucose (C6H12O6). Even more remarkable, all life on Earth uses the same metabolic machinery to extract free energy from glucose not just the sameoverallreactions,butthesamesteps,thesameintermediates,andthesamecontrolling enzymes.Theonlydifferenceisthatnoteveryorganismusestheentirescheme. Itcannotbeemphasizedtoostronglythatthischapterisnotintendedtobeanexercisein memorization. The focus is the pathways of energy flow that living organisms (and specifically microorganisms) use to stay alive. It is far less important that students remember how to write the conversion of one molecule into another, than for them to look at the two molecules, understand what happened between one and other, and recognizehowenergywasliberated.Ourpurposeistoindicatehowaseriesofchemical reactions can be said to have a strategy. The specific strategy utilized specifies the direction in which electrons will travel. In the MFC, we can exploit these strategies and directtheflowofelectrons(energy)forourpurposes.

THESTAGESOFMETABOLISM Metabolismisalwaysinitiatedbythepathwaycalledglycolysis.Glycolysisreleasesonlyasmall amountofenergy.Ifoxygenispresent,glycolysisleadstotwootherpathwaysthatreleasea greatdealmoreenergy.Ifoxygenisnotpresent,however,glycolysisisfollowedbydifferent pathways. METABOLISMWITHOXYGEN=CELLULARRESPIRATION Inthepresenceofoxygen,glycolysisisfollowedbytheKrebscycleandtheelectrontransport chain. When linked together, these pathways make up a process called cellular respiration.

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Cellularrespirationistheoxidationofglucose(C6H12O6)toCO2andthereductionofoxygento water.Thesummaryequationforcellrespirationis: C6H12O6+6O26CO2+6H2O+energy Donotbemisledbythesimplicityofthisequation.Ifcellularrespirationtookplaceinjustone step,alloftheenergyfromglucosewouldbereleasedatonce,andmostofitwouldbeinthe formoflightandheat(justlikeinthefruitsnackdemo).Thekeyforalivingcellistocontrol that energy. It cannot simply start a fire, but must release the explosive chemical energy in sugarsalittlebitatatime.Thecellthenneedstotrapthoselittlebitsofenergybyusingthem toproduceATP,the"molecularcurrency"ofintracellularenergytransfer. QuestionsforReview Whatiscellularrespiration? Cellular respiration is the breakdown of food molecules, such as sugars, into simplercompoundswithareleaseofenergy. Howoftendoescellularrespirationoccurincells? Cellularrespirationoccurscontinuouslyinlivingcells.Ifcellularrespirationstops, acelldies. Createapictureorposterthatshows(a)thewordequationforthecellularrespiration,(b) thechemicalformula,and(c)anonlinguisticrepresentationoftheprocess. C6H12O6+6O26CO2+6H2O+energy glucose+oxygencarbondioxide+water+energy STEP1GLYCOLYSIS Thefirststepintheoverallschemeofenergyextractioninmostorganismsisglycolysis.Inthe first step, one glucose molecule is degraded (essentially broken in half) to two molecules of pyruvicacid(CH3COCOOH),a3carboncompound,withtheproductionofrelativelylittleATP. Although glycolysis is an energy releasing process, the cell needs to input a small amount of energytogettheprocessstarted.Atthebeginningofthepathway,twomoleculesofATPare used up. In a way, the process of glycolysis is analogous to a savings account at a bank. A personhastodepositmoneyintotheaccounttoearninterest,justasacellmustputtwoATP into the account to earn the interest of additional molecules of ATP. When glycolysis is complete,fourATPmoleculeshavebeenproducedfromeachmoleculeofglucose.Thisgives thecellanetgainoftwoATPmolecules. Inmorecriticalreactionsoftheglycolysispathway,fourhighenergyelectronsareremovedand then passed to an electron carrier called NAD+, or nicotinamide adenine dinucleotide. Each

47

molecule of NAD+ can acquire two electrons; that is, be reduced by two electrons. However, only one proton accompanies the reduction. The other proton produced as two hydrogen atomsareremovedfromthemoleculebeingoxidizedisliberatedintothesurroundingmedium. ForNAD+,thereactionisthus: NAD++2HNADH+H+ Consequently,theoverallreactionthatoccursduringtheprocessofglycolysisisasfollows: C6H12O6+2NAD+2C3H4O3+2NADH+2H++2ATP

PGAL=3carbonintermediate; (Glyceraldehyde3phosphate)

QuestionsforReview

What is the source of the energy used by the cell to split the glucose molecule into pyruvate? TwoATPmoleculesprovidetheenergy. Howmanypyruvatemoleculesareproducedfromeachglucosemolecule? Two WhatisaddedwhenNAD+becomesNADH? Electrons Whataretheendproductsofglycolysis? Pyruvateandenergy

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STEPS2&3KREBSCYCLEANDTHEELECTRONTRANSPORTCHAIN The second step in the machinery of cellular respiration is much more efficient in extracting energy. Pyruvic acid (or pyruvate) enters the Krebs cycle, or citric acid cycle (because citric acidistheproductofthefirstreaction),whereitisbrokendowntoCO2,withhydrogenatoms being used to reduce NAD+ to NADH. Some additional ATP also is made along the way. The NADHfromthecitricacidcycleflowsintothethirdprocess,theelectrontransportchain.Here NADHisreoxidizedtoNAD+andisrecycled.Thehydrogenatomsaretranslocatedoverthecell membrane from "inside" to "outside", establishing a concentration gradient across the membrane, which temporarily stores the energy released in the chemical reactions. This potentialenergy(thefreeenergythatisliberated)isstoredintheformofATP.Thehydrogen atoms ultimately are added to O2 to make water. The overall process the combustion of glucosewithoxygencarriedoutintheseseriesofsmallstepssothatthemaximumamountof energyfromthereactioncanbesavedyields38moleculesofATPpermoleculeofglucose. AEROBICVS.ANAEROBICMETABOLISM Atthispointitbecomesnecessarytoclearupafewmisconceptionsandterminologyerrors that frequently occur. It is not uncommon for the terms anaerobic respiration and fermentationtobeusedinterchangeably,butthisisincorrect.Fermentationisthemetabolic process that occurs in microorganisms (and occasionally in human muscle tissue) in the absenceofoxygen.Duringfermentation,thepyruvategeneratedbyglycolysisgoesthrough a series of enzymatic transformations to form reduced metabolites, such as lactic acid or ethanol. During anaerobic respiration, glycolysis is followed by the Krebs cycle and the electron transport chain, just as in normal aerobic respiration, however, the final electron acceptorisamoleculeotherthanoxygen. Anaerobicrespirationisakeyelementtotheunderstandingandthefunctioningofpractical application microbial fuel cells (see Chapter 2 Anaerobic Respiration). Fermentation is a criticalcomponentofoursimpleexperimentalMFC. Under nonstrenuous conditions, when the oxygen supply is ample, glucose is degraded to pyruvateduringglycolysis,andpyruvateisbrokendowntoCO2intheKrebscycleandelectrons transferredtooxygenintherespirationchain,withayieldof38moleculesofATPpermolecule ofglucose.However,duringvigorousexercisewhereO2demandisveryhighinthemuscles,an interesting reaction called fermentation is responsible for allowing our muscles to keep workinghardevenwhenoxygenstartstorunlow. As the cell supplies large amounts of ATP (energy) from glycolysis to muscle cells during exercise,itrunsintoaproblem.Injustafewseconds,allofthecellsavailableNAD+molecules arefilledupwithelectrons(becomingNADH).TheNADHcannotbereoxidizedbacktoNAD+ fast enough since the oxygen, which functions as the final electron acceptor in the electron transport chain, is limited. Without NAD+, the cell cannot keep glycolysis going, and ATP productionstops.Duringfermentationinhumanmuscle,insteadofenteringtheKrebscycle, 49

thepyruvategeneratedbyglycolysisisreducedtolactate.ThisactionconvertsalltheNADH backintotheelectroncarrierNAD+,allowingglycolysistocontinue:
3

O 2CH CCOOH + 2NADH + 2H

pyruvic acid
+

OH 2CH CHCOOH + 2NAD

3 +

lactic acid

Thisisaredoxreactionthatconsistsoftwohalfreactions: (1) (2) pyruvate+2H++2elactate NADH+H+NAD++2H++2e pyruvategainse(isreduced) NADHlosese(isoxidized)

TheNAD+thatisgeneratedisthenavailabletobeusedinglycolysisagaintogeneratemoreATP forthemuscles.Thelactate(lacticacid)generatedbythisreactioncausesacidificationofthe blood(loweredpH),whichisbelievedtoberesponsibleforthe"burn"thatyoufeelinmuscles that you worked too hard. Because this fermentation process does not require oxygen, it is saidtobeanaerobic. ADETAILEDLOOKATFERMENTATION Itispossible(orevenroutine)forotherorganismstoalsoproduceenergywithoutusingoxygen toburnglucose.In1861,LouisPasteurdiscoveredthatyeastcellsarecapableofconverting glucosetocarbondioxideandethylalcohol(ethanol).Thisreactionyieldsonlyabout1/20the amount of energy released when glucose is metabolized all the way to carbon dioxide and water through the use of oxygen (aerobic respiration). This is not a significant problem, however,asyeastdonothaveashighanenergyrequirementasmorecomplexorganisms.The foodindustrymakesuseofthis(mostdelightful)reactiontoproduceavarietyofconsumable (andquitetasty)products. Ethanol fermentation (performed by yeast and some types of bacteria) breaks the pyruvate down into ethanol and carbon dioxide. It is important in bread making,brewing,andwinemaking.Usuallyonlyoneoftheproductsisdesired; inbreadthealcoholisbakedout,andinalcoholproductionthecarbondioxideis releasedintotheatmosphere.

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Lacticacidfermentationbreaksdownthepyruvateintolacticacid.Aspreviously discussed,itoccursinthemusclesofanimalswhentheyneedenergyfasterthan thebloodcansupplyoxygen.However,italsooccursinsomebacteriaandsome fungi. It is this type of bacteria that convert lactose into lactic acid in yogurt, givingititssourtaste. Ethanolandlacticacidaretypicalexamplesoffermentationproducts.However,moreexotic compounds can be produced by fermentation, such as molecular hydrogen, butyric acid, and acetone. For living cells,the productsproducedbyfermentationareactually wasteproducts createdduringthereductionofpyruvatetoregenerateNAD+intheabsenceofoxygen. In our experimental MFC, we will use the waste products of anaerobic bacterial fermentation as a source of electrons. Molecular hydrogen will be oxidized to generate electronsandprotons(H+)inthefollowingreaction: H22H++2e The electrons are subsequently transported to the anode and dropped into the electrical circuitbytheactionofasolubleelectrontransportmediatorcalledmethyleneblue. Humans can use anaerobic respiration and fermentation in much the same way yeast do, including using the same pathways. However, unlike yeast that can excrete alcohol into the surrounding medium, our cells (if we possessed the same enzymes) would have to excrete alcoholintoourbloodstream.Thenwewouldhavetoeliminateitormetabolizeitquicklyto avoidbeingcontinuouslydrunk.Instead,ourbodies,lackingthenecessaryenzymestoproduce alcohol,followanalternativepathway,whichyieldsasubstancecalledlacticacid.Whilelactic acidislesstoxicthanalcohol,itnonethelesscannotbetoleratedattoohighaconcentration. As muscle activity increases, we need more energy, so more glucose is metabolized. As we begintouseoxygen(foraerobicrespiration)ataratethatexceedswhatourlungscanprovide, locallyinthemusclecellsaswitchtothefermentationpathwayoccursandlacticacidquickly buildsup.WerapidlybecomefatiguedandourmusclesstarttoacheandburnasthebloodpH lowersinresponsetotheincreasingconcentrationoflacticacid.Iftheproductionoflacticacid wouldcontinue,wewouldsoonbeunabletomoveatall.Aswelieinaheapontheground pantingforair,ourheartandlungsworkespeciallyhardtodeliverasmuchoxygenaspossible to our starved muscle cells. At this point, the lactic acid can be metabolized further by the oxygenthroughtheremainderofthecellularrespirationpathway.Thismeanswegetthefull benefitofalltheenergyreleasedwhenglucosegoestolacticacidandthentocarbondioxide andwater. To summarize, glycolysis carried out in both aerobic and anaerobic conditions produces pyruvate. The difference is that without oxygen, the pyruvate is turned into lactic acid or

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ethanol,and,whenoxygenispresent,thepyruvatecontinueson(throughtheKrebscycleand electrontransport)toeventuallygivecarbondioxideandwater. Activity2.2LacticAcidSitOutChallenge Findalongwall.Havethestudentslineupagainstthewallandplacetheirbacksagainst thewall.Theythenslidedownthewallintoasittingposition,withtheirthighsparallel tothefloorandtheirshinsparalleltothewall.Theobjectiveofthegameistoseewho can maintain that position for the longest time. Soon after being in the position, the studentsfeeltheburnofthelacticacidaccumulation. Analyze the results of who had the longest wall time, and analyze what the burning feelingisandwhyaburningfeelingbecameaproblem.Thewinnersgetacandybar,and thentherecanbemorediscussionaboutwhatwillhappenuponeatingthecandybar.

QuestionsforReview

Describe the role of NAD+ (nicotinamide adenine dinucleotide) in the cell. Write a chemicalequationforthereductionofNAD+. ThemainfunctionofNAD+istoactasanelectroncarrierintheredoxreactions that occur in the cell during metabolism. It is found in two forms: NAD+ is an oxidizing agent [it accepts electrons from other molecules and becomes reduced,thisreactionformsNADH,whichcanthenbeusedasareducingagent todonateelectrons.] NAD++2HNADH+H+ Summarize what happens during fermentation, and then explain the importance of the process(especiallyintermsofNAD+). Duringstrenuousexercise(inhumans)orinorganismthatexistunderanaerobic conditions, the oxygen debt makes it impossible for the pyruvate generated by glycolysis to be fully metabolized to carbon dioxide, water, and ATP. Subsequently,allNAD+moleculesinthecellbecomesaturatedwithelectronsand areconvertedtoNADH.Withouttheaerobiccellularrespirationprocessesofthe KrebscycleandelectrontransporttorecycleNADHtoNAD+,glycolysisandthe productionofATPwouldcease.Instead,underanaerobicconditions,organisms make a switch and followup glycolysis with fermentation. In fermentation, which does not require oxygen, the pyruvic acid is reduced to lactic acid (in humans)orethanol(inyeastsandsomebacteria),andtheelectronsgenerated are used to oxidize NADH back to NAD+. Thus, NAD+ is again available for glycolysistocontinue.

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Activity2.3.AFermentationbyYeast Students will measure CO2 production by fermenting yeast. Then demonstrate how magnesium ions and compounds containing fluoride enhance, inhibit, or slow enzyme activity.

Activity2.3.AFermentationbyYeast

During the fermentation process by yeast cells, ethanol is produced and carbon dioxide is released.Thechemicalformulaforfermentationinyeastis: Chemicalequation C6H12O62CH3CH2OH+2CO2+2ATP Wordequation Many of the biochemical reactions in the fermentation of glucose require magnesium ions. Magnesium ions activate several enzymes involved in fermentation. To test the effect of magnesium ions on the breakdown of glucose, extra magnesium ions may be added to the reaction.Magnesiumionsmayberemovedfromthereactionsolutionbyaddingacompound containingfluoride.Fluoridecausesmagnesiumionstoprecipitatefromsolutions. Studentsshouldformulateahypothesisforthelabbeforeconductingtheexperiment.[Ifextra magnesiumionsareaddedtothereactionsolution,thenfermentationshouldoccuratafaster rate; if removed, fermentation will slow down.] Review allsafetyprecautionsassociatedwiththislab.Becareful whenusingsodiumfluoride.Itcanbetoxicifingestedor inhaled, and may irritate the skin. If any spill occurs on skinorclothes,itshouldbewashedoffimmediatelywith large amounts of water. Develop a plan to dispose of wastematerialsandinformstudentsoftheplanbefore executingtheexperiment. Organize the class into groups of four students. Work with the class to design the experimental procedures. Inform the groups that they will have available the followingmaterials:five50mLbeakers,greasepencilsor Figure2.1.Fermentationtube sharpies, five fermentation tubes (Figure 2.1 or similar (http://www.enasco.com/product/ gascollectionapparatus),six10mLgraduatedcylinders, SA08681M) yeastsuspension*,distilledwater,10%glucosesolution, Sugar(glucose)alcohol(ethanol)+carbondioxide+energy(ATP)

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0.01Mmagnesiumsulfatesolution,0.06Msodiumfluoridesolution,0.20Msodiumfluoride solution,andametricruler. * To prepare the yeast suspension, mix the entire contents of one fresh package of bakers yeastin90mLofdiH2Oand10mLglucosesolution.Then,incubatethesuspensionat37oCfor 12hourstodeveloptheyeastcolonies. Guidethestudentssothateachbeakercontainsthefollowingmaterials: Beaker#110mLyeastsuspension,20mLdistilledwater Beaker#210mLyeastsuspension,10mLdistilledwater,10mLglucose Beaker#310mLyeastsuspension,10mLglucose,10mLof0.10Mmagnesiumsulfate Beaker#410mLyeastsuspension,10mLglucose,10mLof0.06Msodiumfluoride Beaker#510mLyeastsuspension,10mLglucose,10mLof0.20Msodiumfluoride

Havestudentspourthecontentsofeachbeakerintoafermentationtubenumberedthesame asthebeaker.Afterthegroupshavesetuptheexperiment,allowittorunovernight.Thenext day(Day2),writethefollowingclassdatachartonthechalkboard:

Group#1 Group#2 Group#3 Group#4 Group#5 Group#6 Group#7,etc. Avg.mLof CO2produced Ranking

Tube1 5

CO2ProductionYeast(mL) Tube2 Tube3 Tube4 2 1 3

Tube5 4

SpecialNote:Theresultsforrankingarebasedontheexpectedoutcomeifthetubescontain thematerialsaslistedaboveineachnumberedbeakerandtransferredtothetubewiththe correspondingnumber.

OnDay2,havestudentsmeasuretheamountofcarbondioxidecollectedusingametricruler orreadthevolumeofcarbondioxide(inmL)producedifthefermentationtubeisgraduated. Havestudentsusearatingscaleforeachofthetubes.Use#1forthetubethatproducedthe greatest average amount of carbon dioxide and #2 for the tube with the 2nd largest average amountofcarbondioxide.Continuethisrankingmethoduntilthetubewithnocarbondioxide producedisrated#5.Theexpectedoutcomebasedonthecontentsofthefermentationtubes as listed above is: Tube three should be #1 with the greatest amount of carbon dioxide produced, Tube two should be #2 with the 2nd greatest carbon dioxide produced, Tube four

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should be #3, Tube five should be #4, and Tube one should be #5 with no carbon dioxide produced. Assigneachgroupanumberandhaveamemberofeachgroupwritetheirresultsintheclass datachart.Havestudentscomparetheirresultswiththoseofothergroupsusingtheclassdata chart.Lookforcommonresponses.Iftheclassdatadonotagree,askstudentstoaccountfor anydifferences.Cometoanagreementontherankingforeachtube. AnotherapplicationtothislabwouldbetouseTESTAPE(availableatmostpharmaciesfor$3 $5)tomeasuretherelativeamountofglucoseinthetubesbeforeandafterincubation.TES TAPEisaglucosetestingstripusedbydiabeticstomonitortheamountofbloodsugarinurine, but it will also work to visually confirm that the amount of glucose in the tubes has been reducedovernightastheyeastmetabolizeitintoCO2. Activity2.3.B(Extention)OtherVariablesinFermentation Havestudentsdesignandcarryoutaninquiryexperimenttotestahypothesisaboutthe effects of sweeteners (instead of, and/or in addition to, glucose) on carbon dioxide productionbyyeast.Studentsshouldproposeahypothesisabouthowdifferenttypesand amountsofsweetenerswillaffecttheamountofcarbondioxideproductionbyyeast.Have availablevarioussweeteners,suchastablesugar,cornsyrup,molasses,honey,fruitjuice concentrate, saccharine, aspartame, etc. Remind students to set up a control. Have studentsanalyzethedataanddrawconclusions.Discusstheoutcomeandresults.

ANAEROBICRESPIRATION Certain bacteria, which thrive in chemicallyrich but nearly oxygenfree environments, are adaptedtousesubstancesotherthanoxygenasterminalelectronacceptorsduringrespiration. In anaerobic respiration, as the electrons from the electron donor (e.g., sugars and other organic compounds) are transported down the electron transport chain to the terminal electronacceptor,protonsaretranslocatedoverthecellmembranefrom"inside"to"outside", establishing a concentration gradient across the membrane, which temporarily stores the energyreleasedinthechemicalreactions.ThispotentialenergyisthenconvertedintoATPby the same enzyme used during aerobic respiration. If given a choice, oxygen is always the preferred final electron acceptor, however, other possible electron acceptors (electron sinks) for anaerobic respiration include NO3, SO42, Fe+3, and MnO2. Anaerobic respiration is a commonoccurrenceinnaturebutisrestrictedtoprokaryoticorganisms(e.g.,bacteria). The property of specific bacterial genera (e.g., Geobacter and Shewanella) that makes the microbial fuel cell without artificial mediators, such as methylene blue, possible is their ability to perform anaerobic respiration and use an external electron receptor, as opposed to fermentation, which utilizes the internally generated electron acceptor, pyruvate. When no dissolved electron acceptors are available, these bacteria are able to transport electrons to extracellular undissolved electron acceptors, such as some metals and electrodes (i.e., anodes), and thus create an external

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voltage potential between the solution and the undissolved electron acceptors (i.e., the anode). Another critical adaptation of these bacteria is that in the absence of soluble electron acceptors, they automatically colonize and grow to cover the electrode that is acting as the electron acceptor (see Chapter 3 Bacterial Biofilms).

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Chapter3:WhatisaMicrobialFuelCell?

PRINCIPLESOFFUELCELLS Afuelcellisanelectrochemicaldevicecapableofthedirectconversionofchemicalenergyinto electricalenergy.Itproduceselectricityfromanexternalfuel(ontheanodeside)fromwhich electronsarewithdrawn,andthentransferredtoanoxidant(onthecathodeside).Thecircuit isclosedbyanionorprotonexchangeconnectionbetweenthefuelandoxidantchamber.Fuel cellsaredifferentfrombatteriesinthattheyconsumereactant,whichmustbeconstantlyfed intothedevice,whereas,batteriesstoreelectricalenergychemicallyinaclosedsystem.Fuel cellscanoperatevirtuallycontinuouslyaslongasthenecessaryflowsaremaintained.Many combinationsoffuelandoxidantarepossibleinafuelcell.Ahydrogenfuelcelluseshydrogen asfuelandoxygenasoxidant(Figure3.1).

Figure3.1.Abasicchemical(orhydrogen)fuelcell Inessence,afuelcellworksbycatalyticallyseparatingthecomponentelectronsandprotonsof thereactantfuel,andthenbyforcingtheelectronstotravelthroughacircuit,convertingthem toelectricalpower.Theprotonsmovethroughaseparator(ionconductivemembrane)tothe cathode to maintain electroneutrality. The catalyst is typically comprised of a platinum group metal, or alloy, that has been coated onto a carbon or graphite electrode. Another catalytic processtakestheelectronsbackin,combiningthemwiththeprotonsandtheoxidanttoform wasteproducts(typicallysimplecompoundslikewaterandcarbondioxide).

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The effectiveness of the electron transfer at the electrode surface, and subsequent fuel oxidationattheanodeandthereductionoftheoxidantatthecathodesurface,aredetermined bytheeffectivenessofthecatalyst.Thedifferencesinthestandardelectrodepotentialsforthe anodicoxidationofthefuelandforthecathodicreductionoftheelectronacceptordetermine thecontentofstoredchemicalenergyinthefuelcell.Thelargerthedifferencebetweenthe individualelectrodepotentials,themoreelectricalenergycanbegenerated(seeChapter1 StandardReductionPotentials). Todeliverthedesiredamountofenergy,thefuelcellscanbecombinedinseriesandparallel circuits,whereseriesyieldshighervoltage,andparallelallowsastrongercurrenttobedrawn (seeChapter1ElectricalCircuits).Suchadesigniscalledafuelcellstack.

Whatisthechiefdifferencebetweenabatteryandafuelcell? Batteries have a definite fuel stock, which will eventually become depleted and the battery will be dead. Fuel cells can run indefinitely because fuel is constantlyfedintothedevice.

PRINCIPLESOFBIOFUELCELLS In a biofuel cell, electrons are made accessible from a nonelectroactive fuel by the use of biocatalysts.Biofuelcellscanbecharacterizedaseitherenzymaticfuelcellsormicrobialfuel cells(MFCs),dependingonthekindofbiocatalystusedenzymesorbacteria,respectively. EnzymaticFuelCells Enzymatic fuel cells usually employ immobilized enzymes (commonly redox enzymes) as catalysts to accelerate highly specific reactions. Purely enzymatic fuel cells have the great advantage of being very small in scale. Due to the small size of the enzymes and the high specificity of the anode and cathode reactions, they have high turnover rates and, thus, high power densities (and a membrane separation of the electrodes is often not necessary). Therefore, they give the possibility to construct low energy power supply units for small electricaldevicesaswellaspossibleinvivoapplications(e.g.,medicalimplantations).

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Activity3.1CanaFuelCellRunonCoke?

Havethestudentsreadthearticlesfoundat: http://blog.wired.com/gadgets/2007/03/like_programmer.html http://www.slu.edu/x14605.xml http://www.slu.edu/readstory/more/4479 http://www.slu.edu/readstory/newsinfo/2474 Have students discuss how an enzymatic fuel cell functions. After reading the articles, discusstheadvantagesanddisadvantagesofenzymaticfuelcelltechnology.Doyouthink thistechnologyhasanypracticalapplicationsinthenearfuture?Whyorwhynot?

MicrobialFuelCells Microbial fuel cells (MFCs) are as diverse chemically as the bacteria that power them. In an MFC,theoxidationreactionsoccurinsidethebacteria,andelectronsmustthenbetransferred totheextracellularanode.Thoughtheyoperateonthesameprinciples,thedesigngoalsofthe MFCaredifferentfromthoseofachemicalfuelcell.Ratherthananonrenewablesource,such ashydrogengas,MFCsusebiomassasthesubstrate(metabolicoxidationreductionreactions) andmicroorganismsasthecatalyst,exploitingwholelivingcellsinanaimtogainenergy. Describetheoxidationandreductionreactionsthatoccurinbacteria. Bacteria oxidize glucose during the process of glycolysis, producing two, three carbonmoleculesofpyruvate.Underaerobicconditions,thepyruvatecontinues through the pathways of the Krebs cycle and electron transport to eventually yieldH2Oasmolecular oxygen(O2)isreduced.Underanaerobicconditions(no O2),thepyruvateundergoesfermentationtoyieldeitherethanolorlacticacidor anaerobicrespiration(seeChapter2AerobicRespirationvs.Fermentation). BIOLOGICALPRINCIPLESOFMFCs In terms of the chemistry, every biological degradation of organic matter is an oxidation process.However,whenwekeepthedegradationanaerobic,wegetthechancetoexploitthis process for electron recovery (power production). Anaerobic conversion of sugars is realized either by bacterial fermentation leading to the formation of small, reduced energyrich metabolic products, such as ethanol, acetate, or hydrogen or by anaerobic respiration using anotherterminalelectronacceptorinsteadofoxygentotakeuptheelectronscomingfromthe sugar. Different MFC techniques allow us to utilize both of these anaerobic metabolic processes. 59

Whatmustbedonetoensureananaerobicenvironmentintheanodechamberofthe MFC? Wemustnotallowoxygenintothechamber.(Note:Studentsshouldbeinformed thatitwillbenearlyimpossibletokeepthechambercompletelyanaerobicdueto theopenings(ports)intheMFCbodies.)Thebacteriawillbegrowninasealed container,sonooxygencangetin.Whenthebacterialsolutionisaddedtothe MFC,itmustbedonequicklytolimitoxygenexposure. AtypicalMFCconsistsoftwoseparatechambers,whichcanbeinoculatedwithliquidmedia (Figure3.2).Thesechambers,ananaerobicanodechamberandanaerobiccathodechamber, aregenerallyseparatedbyanionexchangemembrane.AMFCsuchasthiscanbeclassified intotwotypes.Onetypegenerateselectricityfromtheadditionofartificialelectronshuttles (mediators)toaccomplishelectrontransferfrombacterialcytoplasmtotheanode.Theother typedoesnotrequiretheseadditionsofexogenouschemicalsandcanbelooselydefinedasa mediatorlessMFC.MediatorlessMFCscanbeconsideredtohavemorecommercialpotential then MFCs that require mediators because the typical mediators are expensive. One major challenge is that, if oxygen will be used as the final electron acceptor, the cathode chamber needstobefilledwithasolutionandaeratedtoprovideampleoxygentothecathode.

Figure3.2.Abasicmicrobialfuelcell
(reprintedwithpermissionofCellPress)

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ELECTRICITYGENERATIONINTHEMFC In normal microbial metabolism (see Chapter 2), a carbohydrate (glucose) is oxidized initially without the participation of oxygen when its electrons are released by enzymatic reactions. Theelectronsarestoredasintermediates,whichbecomereduced,and,inthisstate,theyare used to fuel the reactions, which provide the living cell with energy for maintenance and growth.Theultimateelectronsink(orendpointoftheredoxreaction)ismolecularoxygen (dioxygen,O2).Toanelectrochemist,asimplifiedrepresentationoftheanodehalfcellreaction involvedintheoxidationofglucosebyawholebacterialcellwouldbeasfollows: C6H12O6+6H2O6CO2+24H++24e The large harvest of electrons is stored as reduced intermediates, but the eventual terminus therespiratorychainisoxygen,asdemonstratedinthecathodehalfcellreaction: 6O2+24H++24e12H2O

pHEFFECTSINTHEMFC The term pH refers to the concentration of hydrogen ions (H+) in a solution. An acidic environmentisenrichedinhydrogenions,whereas,abasicenvironmentisrelativelydepleted of hydrogen ions. The pH of biological systems is an important factor that determines which microorganismsareabletosurviveandoperateinaparticularenvironment,suchastheanode chamberofamicrobialfuelcell.MostmicroorganismspreferpHvaluesthatapproximatethat ofdistilledwater,aneutralsolution. ThehydrogenionconcentrationcanbedeterminedempiricallyandexpressedasthepH.The pHscalerangesfrom0to14,with1beingthemostacidicand14beingthemostbasic.ThepH scale is a logarithmic scale. That is, each division is different from the adjacent divisions by a factorof10.Forexample,asolutionthathasapHof5is10timesasacidicasasolutionwitha pHof6. Therangeofthe14pointpHscaleisenormous.DistilledwaterhasapHof7(neutral).ApHof 0correspondsto10millionmorehydrogenionsperunitvolumeandisthepHofbatteryacid.A pHof14correspondsto110millionthasmanyhydrogenionsperunitvolume,comparedto distilledwater,andisthepHofliquiddraincleaner. Compounds that contribute hydrogen ions to a solution are called acids. For example, hydrochloric acid (HCl) is a strong acid. This means that the compounds dissociate easily in solutiontoproducetheionsthatcomprisethecompound(H+andCl).Thehydrogenionisalso

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aproton.Themoreprotonsthereareinasolution,thegreatertheacidityofthesolution,and thelowerthepH. Mathematically,pHiscalculatedasthenegativelogarithmofthehydrogenionconcentration. Forexample,thehydrogenionconcentrationofdistilledwateris107and,hence,purewater hasapHof7. pH=log[H+]=log107=7 In the MFC, maintaining the pH of microbial anode chamber and the anode feed solution is importanttoensurethatgrowthofthetargetmicrobesoccurs.Also,pHneedstobemonitored astheanodeandcathodereactionsareoccurring.IfthepHvariestoowidely,thegrowthand metabolismofthemicroorganismcanbehalted.Thisinhibitionisduetoanumbersofreasons, such as the change in shape of proteins due to the presence of more hydrogen ions. If the alteredproteinceasestoperformavitalfunction,thenormalfunctioningandeventhesurvival ofthemicroorganismcanbethreatened. Ion exchange membranes pose an important challenge in pH maintenance in the MFC. Ion exchangemembranesseparatethebiologicalanodefromthecathodereactions,while,atthe same time, facilitating the transport of ions through the membrane to maintain electro neutralityinthesystemandproperbacterialrespiration.Iontransferbetweentheanodeand cathode isnecessarybecause ofthemovementofnegativelychargedelectronsfromthe anode to the cathode. To achieve a counterbalance, either negative charge equivalents (anions/hydroxide ions) travel from the cathode to the anode, or positive charge equivalents (cations/protons)movefromtheanodetothecathode, dependingontheselectionoftheion exchangemembranematerial.

Since the cathode reactions of MFCs consume protons in equal amounts as electrons, ideally only protons are transported through the ion exchange membrane. In this way, electro neutralityisobservedwithoutpHchangestakingplaceatthecathode.However,becauseMFCs operate near neutral pH in the anode and cathode chambers, the concentration of cations otherthanprotons(e.g.,Na+,K+,NH4+)aretypically105timeshigherthanH+ionsinsolution. ThiscompetitivetransportofcationsotherthanprotonssignificantlyeffectsMFCperformance. When the substrate is degraded (metabolized), protons are produced at the anode and consumed at the cathode. However, if due to competition and concentrations gradients, protonscannotmigrateatasufficientratefromtheanodetothecathode,thepHwilldecrease attheanodeandincreaseatthecathodewhilechargebalanceismaintainedbythemigration ofothercations.ThepHdecreaseattheanodeaffectsbacterialrespirationand,thus,current generation.UtilizingwellbufferedsolutionsintheMFCcanoffsetthesepHchanges,butthe addition of chemical buffer solutions is not sustainable and it is not clear to what extent localizedpHchangesintheMFCmayaffectpowergeneration.

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At this point, it may be necessary to explain how certain bacteria can transfer electrons directly to an electrode and why, in some instances, mediators are necessary. Since electrodesaresolidentitiesthatcannotpenetratethebacterialcells,amajorrequirementis that electrons are to be transferred from the inside of the microbial cell membrane to its outside either via the physical transfer of reduced compounds, or via electron hopping acrossthemembraneusingmembraneboundredoxenzymes.Ananalogyofeatingacookie can be used to explore how the process works. When we eat a cookie, the sugars in the cookie are acting as electron donors as they are oxidized. The electron acceptor, then, is oxygen.Studentsshouldunderstandthatthesourceofoxygenistheairaroundthemasthey respire. As they breathe in oxygen, it is available to be used as an electron acceptor. It is more difficult, though, to use the same analogy for bacteria that are transmitting electrons directly to an electrode. The electron donor is again sugar (glucose), but how can an electrodebebreathedin?Howcantheelectronsbeinggeneratedbythereactionsinside the cell be transported across cell membranes to the electrode? These questions will be answeredtoanextentinthefollowingsections,butitshouldbenotedthatthisisabudding area of MFC research and not all of the mechanisms of direct electron transfer to the electrodeareknownorfullyunderstood. MEDIATORLESSMFCs (1)Theuseofbacterialbiofilms BacterialbiofilmMFCsarebasedonthedirectphysicalandelectronicinteractionbetweenthe microorganisms and the anode surface (Figure 3.3). A biofilm is a community of microorganismsadheringtoasurface.InthecaseofMFCtechnology,thesurfaceistheanode (i.e.,electronacceptingelectrodeseeChapter2AnaerobicRespiration).Therearecurrently threemaintheoriestowhichmostMFCresearcherssubscribe:(1)Thebacteriaadheringtothe anode have electrochemically active redox enzymes on their outer membranes that contain iron molecules. The iron molecules orient themselves in such a way as to traverse the outer membraneofthebacterium,allowingthedirecttransferofelectronstoexternalmaterialsand, therefore, do not require any chemical assistance to accomplish electron transfer to the electrode; (2) Some evidence suggests that electrons may be transferred directly to the electrodeviaconductive,hairlikeproteinappendages(orpilli)foundonthesurfaceofcertain bacterialgenera;and(3)Certainbacterialspeciescansynthesizemicromolaramountsoftheir ownredoxmediatorsthatcanthenbeusedbyawholerangeofbacteriainamixedculture. Themethodologynotwithstanding,whenthesebacteriaoxidizetheorganicmatterpresentin thesubstrate,theelectronsareshuttledtotheanode.Oxygen,thehydrogenprotons,andthe electronsthatareconnectedbyacircuitfromtheanodetothecathode,arethencatalytically combinedwith(routinely)aplatinumcatalysttoformwater(Figure3.3).(Platinumisacritical

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catalystatthecathodeandnoalternativemetalhasbeenproventocatalyzethecombination ofoxygen,thehydrogenproton,andtheelectroninamoreefficientmanner.) In MFC research, microbial biofilms are quite common, however the startup phase of such biofilmbasedMFCanodesnormallytakesdaysorweeksand,therefore,cannotberealizedin shortterminvestigations(butmaybesuitableforalongtermschoolproject).

Figure3.3.BacteriafixedinabiofilmatanMFCanode (2)Directoxidationofsecondaryfuelsattheanode Asecond,highlypromisingMFCapproachisbasedonthepreparationandinvestigation ofelectrocatalyticanodes,capableofanefficientanddirectoxidationofbacterialend productsunderthediverseandcomplexmicrobialgrowthconditions(seeChapter2 AerobicRespirationvs.Fermentation).Again,platinumcanbeusedasacatalystforthe utilization of molecular hydrogen by fermentative microorganisms at the anode. However,atthisstage,thelowstabilityandhighcostsofplatinumpreventlargescale applications.

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Figure3.4.Transformationofaprimarysubstrateintoasecondaryfuelfordirect oxidationattheanode(Ptorothercatalystrequired)

PlatinumcoatedcarbonelectrodescouldbeusedasboththeanodeandcathodeinourMFC (Figure3.4).Attheanode,platinumwouldfunctionasanexcellentelectronacceptorduring the oxidation of molecular hydrogen, H2, present as a byproduct of the initial anaerobic processing of glucose. In the cathode, platinum is capable of donating electrons during oxygen reduction to produce water. There are several reasons we have chosen not to use platinum electrodes in our experimental MFC. (1) Platinum electrodes are very easily poisoned by the products of microbial metabolism if they are not coated with a protective polymer prior to use in the MFC. Sulfides, carbon monoxide, and, to a lesser extent, even carbondioxidepresentintheanodechambercancausethemoleculestobindtightlytothe surfaceoftheplatinumelectrode,renderingituselessforelectrontransport.Theplatinum and polymer coating is quite expensive and the preparation of the electrodes is a very detailed process requiring specific equipment and computer software that does not lend itself to simple application in a high school setting. (2) The reduction of oxygen at a platinumcoatedcathode,thoughnotunworkable,doesfurthercomplicatethesetupofthe MFC.Anaquariumpump(orsimilarapparatus)mustbeusedtoprovideaconstantsourceof oxygenintothecathodechamber.Thiscanbeaccomplishedbyattachingathinneedletothe tubingofthepumpandtheninsertingtheneedleintothetopportofthecathodechamber.

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THEROLEOFMEDIATORS Direct electron transfer from most bacteria to an electrode is hampered by overpotentials, whichcanbedescribedastransferresistances.Toreducetheseresistances,thesurfaceareaof the electrodes needs to be increased, and/or redox mediators need to be added to the solution.Aredoxmediatorisacompoundthatcanbereversiblyoxidizedorreduced.Bacteria can use redox mediators to deposit their electrons onto an electron acceptor they cannot directlyreduce(Figure3.5). In the absence of oxygen, electrons may be diverted from the respiratory chain by a redox mediator, which enters the outer cell membrane, becomes reduced, and leaves again in the reduced state. The reduced mediator then shuttles the stolen electrons to the anode. To completethecircuit,asecond(oxidizing)electrode(thecathode)isrequired,againfunctioning astheelectronsink.Theoxidizingmaterialcanagainbeoxygengas,butismoreconvenientfor schoolpurposestouseasimplesolubleoxidizingagent,suchaspotassiumferricyanide. In early MFC research, chemical redoxmediators (often dyes, such as methylene blue or neutral red) were used to shuttle metabolic energy (in the form of electrons) from the cytoplasmofbacteriatoananode.Today,theseexpensiveandtoxicdyesnolongerplayarole forthedevelopmentofpracticalMFCs,mainlybecauseincontinuoussystems,thesemediators would have to be added and recycled permanently. However, these mediatorassisted MFCs arestillaverygoodacademicdevicetostudyelectrontransferprocessesofmicroorganisms.

Figure3.5.OurexperimentalMFCdesignusingamethyleneblueredoxmediatorin theanodeandferricyanideinthecathode

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Activity3.2MethyleneBlueDyeReductionTest Students will conduct a simple experiment to show that anaerobically active bacteria collectedfromthesoilhavethecapabilityofreducingmethyleneblue.

Activity3.2MethyleneBlueDyeReductionTest
INTRODUCTION Though it has lost popularity with the advent of new technologies, the methylene blue dye reductiontestremainsafairlyaccurateindicatorofthebacterialcontentofmilk.Thetestis basedonthefactthatthecolorimpartedtomilkbytheadditionofadye,suchasmethylene bluewilldisappearmoreorlessquicklydependingonthenumberofbacterialcellspresent.The removal of the oxygen from milk and the formation of reducing substances during bacterial metabolismcausesthecolortodisappear.Theagentsresponsiblefortheoxygenconsumption are the bacteria. Though certain species of bacteria have considerably more influence than others,itisgenerallyassumedthatthegreaterthenumberofbacteriainmilk,thequickerthe oxygenwillbeconsumed,andinturn,thesoonerthebluecolorwilldisappear.Thus,thetime ofreductionistakenasameasureofthenumberoforganismsinmilk. Thedairyindustryclassifies(grades)themilkbasedonbacterialcontent: Class1.Excellent,notdecolorizedin8hours. Class2.Good,decolorizedinlessthan8hours,butnotlessthan6hours. Class3.Fair,decolorizedinlessthan6hours,butnotlessthan2hours. Class4.Poor,decolorizedinlessthan2hours. In this activity, we will use the methylene blue reduction test to qualitatively assess our anaerobic bacterial cultures (which will later be used in our experimental MFC) ability to reducemethyleneblue.Thiswillvisuallyconfirmthatmethyleneblue,whenintroducedinto the anode chamber of our MFC, will be capable of mediating the transfer of electrons from insideofthebacterialcellmembranetotheanode.

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MATERIALS Fertile soil (free of chemicals), anaerobic bacterial growth medium, 10 mM methylene blue solution,solutionbottleswithcaps,glassPetridish,oven,15mLconicaltubes,30oC(optimal growthtemperatureforClostridium)waterbathapparatusorincubator,pipettes,pipettorsand tips PROCEDURE Preparationstobedoneatleastonedayinadvanceofexperiment: Preparationofrequiredsolutions o Anaerobicbacterialgrowthmedium For1LofdiH2Oadd: - 2.0g NH4HCO3 - 3.6g KH2PO4 - 0.1g MgSO4x7H2O - 0.01gNaCl - 0.01gNa2MoO4x2H2O - 0.01gCaCl2x2H2O - 0.015gMnSO4x7H2O - 0.00278gFeCl2 - 2.0g yeastextract - 5.0g glucose AdjustpHto6.50.5andautoclave(alternatively,forschoollab,usefreshly boiled,cooleddownwaterforpreparationtomaintainsterilityuntiluse,do notletwaterstandopen.). Note:Soilculturemustbegrownovernight(orlonger)tobeactiveduring experiments! o MethyleneBlue(syntheticelectronmediator) - Preparea10mMsolution(0.032g/10mL) Preparationofanaerobicsoilculture 1. Put some fertile soil in a glass Petri dish and heat uncovered in an oven at approximately120Cforonehour. 2. Fillatightlysealablebottlewithbacterialgrowthmedia(about150mL)and addasmallamount(about1tsp)ofheatpretreatedsoil. 3. Grow the bacterial culture overnight at 30C (or longer at room temperature).Thesolutionshouldturncloudyandstarttofoam.Caution:If growinglongerthanoneday,buildupofgaspressurecouldcausebottleto rupture! 68

 Dayofexperiment: 1. Measureincreasingamountsofmethyleneblueintoaseriesof15mLconicaltubes: Tube1nomethyleneblue,MB(control) Tube20.25mLMB Tube30.50mLMB Tube41.0mLMB Tube52.0mLMB Tube61.0mLMB+H2O(negativecontrol) 2. Add1213mL(filltubecompletely)ofbacterialsuspensiontoeachtube15and12 mLwatertotube6;tightlyscrewoncap.Gentlyinverttubeoncetoensuremixingof bacteriawiththemethyleneblue.Donotshaketubes!Shakingintroducesoxygeninto theanaerobiccultures. 3. Immediately place tubes in 30oC incubator or water bath. Record this time as the beginningoftheincubationperiod.Covertubestokeepoutlight. 4. Check samples for decolorization after 15 minutes of incubation. Make subsequent readingsat30minuteintervalsthereafter.Decolorizationisdeterminedbycomparison ofthethemethylenebluecontainingtubestothecontrol(noMB). 5. After each reading, remove decolorized tubes and then slowly make one complete inversionofremainingtubes.Again,noshaking. 6. Recordreduction(decolorization)timeinminutesforeachsample.Forexample,ifthe samplewasstillblueafter15minutesbutwasdecolorizedatthe45minutereading,the reductiontimeshouldberecordedastheaveragetimebetweenthetwoobservations, or30minutes. Factors affecting the test. Many factors affect the methylene blue reduction test, and thereforethestepsofoperationshouldbeuniform. Sincetheoxygencontentmustbeusedupbeforethecolordisappears,anymanipulationthat increasestheoxygenaffectsthetest.Thetubesshouldbecompletelyfilledwiththebacterial solution (even to the point of overflowing). Do not shake the tubes, and mix only by gentle inversion. Light hastens reduction, and therefore the tests should be kept covered. The initial concentrationofthedye(10mM)shouldbeuniformasanincreasedconcentrationlengthens 69

Note: We are working with an environmental sample of microorganisms, some of whichmayalsobepathogenic.Thus,usegloveswhenhandlingthemicrobialsolution andwashyourhandscarefullywhenyouarefinished.Cleanupspillswithbleachand decontaminatesolutionwithbleachattheendofyourexperiment.

the time of reduction. Increasing the incubation temperature augments the activity of the bacteria,andthereforeshortensthereductiontime. Overtimethebacteriawillsettletothebottomofthetube.Thisfactorcausesvariationsinthe reduction time, since the bacteria are not evenly distributed. The accuracy of the test is increased, reduction time shortened, and decolorization more uniform if the samples are periodicallyinvertedduringincubation. How does the methylene blue reduction test show that our soil culture bacteria are active? Ifthebacteriaarecapableofdecolorizingthemethyleneblue,itmeanstheyare actively undergoing anaerobic metabolism. The bacteria are oxidizing the glucoseinthenutrientbrothandgeneratingelectrons.Whenthebacterialcells takeup themethyleneblue intotheir outercellmembrane,thoseelectrons are usedforreducingthedyetothecolorlessformofthemethylenebluemolecule. How can the methylene blue reduction test show that we have an anaerobic bacterial culturecapableofgeneratingelectricityintheMFC? The reduction of the methylene blue we are witnessing is exactly the same reactionthebacteriawillbecarryingoutintheanodecompartmentofourMFC. Only in the MFC, the electrons generated by bacterial metabolism do not stay with the methylene blue molecules. Molecules of methylene blue are back oxidized at the anode and the electrons are subsequently dropped into the electricalcircuitoftheMFC. ADDITIONALDEMONSTRATION At the conclusion of the dye reduction test, pool all of the decolorized tubes into a largersolutionbottle(largeenoughthatthebottleislessthanhalffull).Byswirlingthe contents of the bottle, oxygen trapped in container will become dissolved in the bacterial solution and will backoxidize the methylene blue, returning it to the blue (oxidizedform). Thisis a greatvisual demonstration to use with thestudentssothey canunderstandwhythemethyleneblueretainsitscolorintheMFC.IntheMFC;the methylene blue is only an electron carrier and does not remain in the reduced (colorless) form. As the methylene blue is reduced by electrons generated by fermentation, the electrons are subsequently dropped into (donated to) the electrical circuitattheanode.

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ELECTRONFLOWINTHEMFC Remember that during glycolysis, glucose is broken down into carbon dioxide in an oxidation reaction(seeChapter2Step1Glycolysis).Oxidationreactionsarealwayscharacterizedby alossofelectrons. C6H12O6+6H2O6CO2+24H++24e The electrons released have to be deposited in some sort of electron sink and expelled as waste.Ifthecellcouldnotgetridofitselectrons,itwouldbuildupahugenegativecharge. Frequently,duetoitsavailability,oxygenfunctionsastheelectronsink.Amoleculeofoxygen takesupelectronsandcombineswithprotons(H+)tomakewater. 6O2+24H++24e12H2O The electron sink, however, is not limited to oxygen. During anaerobic respiration, the final electron acceptor could be any molecule capable of accepting electrons provided the proper enzymatic pathway is available. So, how do we know in which direction (and to what molecules)theelectronswillflowinourMFC?Theanswerisrelatedtothereductionpotential of the various electron carriers involved in the metabolic pathways of the bacteria and the mediatorswehaveintroducedintoourMFC. Thetablebelowgivestheapparentbiologicalstandardreductionpotentials(EatpH7,instead ofEatstandardconditions,seeChapterIStandardReductionpotentials)ofsomeimportant biological halfreactions. Electrons flow spontaneously from the more readily oxidized substance (the one with the more negative reduction potential) to the more readily reduced substance (the one with the more positive reduction potential). Therefore, more negative potentials are assigned to reactions that have a greater tendency to donate electrons (i.e., reactionsthattendtooxidizemosteasily).

StandardReductionPotentialsofInterestinMFCs HalfCellReactions StandardReductionPotential(E) + 2H +2e H2 0.42V + + + NAD +2H +2e NADH+H 0.32V Acetylaldehyde+2H++2eEthanol 0.20V + Pyruvate+2H +2e Lactate 0.19V + MB(ox)+2H +2e MB(red) +0.01V 3 4 Fe(CN)6 +e Fe(CN)6 +0.44V NO3+2e+2H+NO2+H2O +0.42V + 2+ MnO2+4e +4H Mn +2H2O +0.60V 3+ 2+ Fe +e Fe +0.76V + O2+2H +2e H2O +0.82V

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It is important to note the direction of all these reactions is always written in the form of a reductionorgainofelectrons.That'snotimportantwhenitcomestodeterminingthedirection ofelectronflow.Forexample,notethatthereductionofprotonstohydrogenisatthetopof the list (Eo = 0.43 V). Electrons released by the oxidation of molecular hydrogen will flow to anyhalfreactionthathasahigher(lessnegative)standardreductionpotential.Inthiscase,the electronsendupinNADH(Eo=0.32V).Noticethereductionofoxygeniswaydownatthe bottomofthelist.That'swhyit'saneffectiveelectronsinkforgettingridofelectrons.Other naturalelectronsinksincludenitrate(NO3),iron(Fe3+),andmanganese(MnO2)(seeChapter2 AnaerobicRespiration).

In our experimental MFC, fermentative bacteria will convert the glucoserich media into mainlybutyrate,acetate,andmolecularhydrogen.Whenthisbacterialsolution,initsredox state, is filled into the anode chamber together with the methylene blue synthetic redox mediator, the mediator itself will be reduced by the bacterial cells, as well as by molecular hydrogen. If the electric circuit to the cathode is closed (that means both electrodes are connected by a wire), the reduced mediator is reversibly oxidized at the anode and its electronsaredroppedintotheelectriccircuit.Bymakingmethyleneblue,whichhasahigher standardreductionpotential,availabletothebacteria,wecandiverttheelectronsawayfrom thenormalpathwaysofethanolorlactatefermentation.Theelectronsthenflowthrough theclosedcircuitfromanodetocathodedue,again,tothedifferencesinelectricalpotential. Thepotassiumferricyanideintroducedintothecathodechambercanthenactastheelectron sinkastheferricyanideionisreducedtotheferrocyanideion.Again,oxygen(ataplatinum coatedelectrode)couldbeusedasthefinalelectronacceptorinourexperimentalMFC,but an apparatus for the continuous delivery of oxygen into the cathode chamber (aquarium pump)isnecessary.

Whateffectwouldusingoxygenasthefinalelectronacceptoratthecathodehaveonthe electricitygeneratingabilitiesoftheMFC? Usingoxygenwould(theoretically)resultinmorepowerproductionintheMFC. Since the oxygen halfreaction has a more positive reduction potential, the potentialdifferencebetweentheoxidationandreductionhalfreactionswouldbe greater.Alargerpotentialdifferencemeanslargervoltageacrossthecircuit(see Chapter1ReductionPotentials).

Activity3.3TheMFCinAction StudentswillconstructtheexperimentalMFC,loadtheanodeandcathodechamberswith theappropriateoxidantandreductant,andharvesttheelectricitygeneratedtopoweran LED. Note:StudentsfrequentlyhavedifficultlyassemblingtheMFC.Sometimeshouldbegiven priortothedayofexperimentationforpractice.Agoodideamaybetoreviewthestep bystep procedure one day prior to the actual date of experimentation, and have the studentsassembleadryMFCusingtheFigures3.6,3.7,3.8,and3.9providedasguides.

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 Activity3.3TheMicrobialFuelCellinAction
MATERIALS TheexperimentwillbeconductedinamodifiedmodelMFCcellcommerciallyavailableatthe NCBE,UniversityofReading,UK(http://www.ncbe.reading.ac.uk/). ThemodelMFCsetscontaintheacrylicMFCbodies,screwstoassemblethem,rubbergaskets, andionexchangemembraneandcarbonclothmaterialforthepreparationofelectrodes. Note:TosetuptheMFCasdescribedinthefollowingprotocol,twocompleteMFCkitsare needed.AssemblingtheMFCwithasingleanodeandcathodechamberwilldemonstratecell potential,butitisnotsignificantenoughtolighttheLED.Ifthedesiredoutcomeistoallow studentstheopportunitytopowertheLED,twoanodechambersmustbeassembled(Figure 3.6andFigure3.7).

Hole through which chamber is filled Neoprene gasket Terminal of carbon fibre electrode Neoprene gasket

Hole through which chamber is filled

Ion exchange membrane

Ion exchange membrane Chamber glued to end plate with Superglue

Figure3.6.The3chamberedMFCinexplosionview

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 Preparationsdoneinadvanceofsession: Presoaktheionexchangemembraneindistilledwaterfor24hourspriortouse. Allelectrodepreparationandmodification o Carbonclothcuttorightsizeforcellcompartments o Pieces of carbon cloth glued to graphite rods* for external connection of electrodesusingconductivecarboncement** o For the preparation of electrodes in the high school (no graphite rods or conductivecementneeded),thecarbonclothcouldbecutinsuchawaythata thin strip of cloth can be led through the connection hole in the acrylic body, providinganexternalconnection(Figure3.7).

Figure3.7.TheassembledMFC
*Graphiterodsarehighcarboncontentpencilrods(verysoft)purchasedfromtheart supplysectionofthecampusbookstore. **Conductivecarboncementisavailableat: http://www.emsdiasum.com/microscopy/products/chemicals/adhesive.aspx#12664

Preparationofrequiredsolutions o Anolytebacterialgrowthmedium For1LofdiH2Oadd: - 2.0g NH4HCO3 - 3.6g KH2PO4 - 0.1g MgSO4x7H2O - 0.01gNaCl

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- 0.01gNa2MoO4x2H2O - 0.01gCaCl2x2H2O - 0.015gMnSO4x7H2O - 0.00278gFeCl2 - 2.0g yeastextract - 5.0g glucose Adjust pH to 6.5 0.5 and autoclave (Alternatively, for school lab, use freshly boiled, cooleddown water for preparation to maintain sterility until use, do notletwaterstandopen.). Note:Soilculturemustbegrownovernight(2436hours)tobeactive during experiments! Bacteria tend to slow metabolism (and their resultingelectroactivity)after48hoursasglucoseisdepleted.

Additionalmaterialsneeded: Two30mLsyringesfordeliveryofsolutionsintoanodeandcathodechambers(Figure 3.9) Glasswareforpreparingandstoringsolutions TubeconnectorsandmatchingtubestoconnectsyringestoMFCcells Fertilesoil(withoutchemicalfertilizers)asasourceofanaerobicbacteria GlassPetridishfortheheatpretreatmentofsoil Handheldmultimetersfortheobservationofcellpotentialandcurrent Largetraytopreventspillsofsolutions Realloads,e.g.LEDlightorsmallfans AlligatorclipsandcopperwireformakingconnectionsbetweenchambersandtoLED Glovesandsafetygoggles 75

o MethyleneBlue(syntheticelectronmediator) - Preparea10mMsolution(0.032g/10mL) o Catholyteferricyanide: - Preparea100mMsolutionofpotassiumferricyanide(8.23g/250mL) CAUTION:Potassiumferricyanideishazardousandshouldnotcomein contact with the eyes. Eye protection is necessary when handling this material.Ifthesolutiondoescomeincontactwiththeeyes,floodthem with water and seek medical attention. Do not let Potassium ferricyanidereactwithstrongacids(poisonformation).Followdisposal guidelinesattheendofthisprotocol.

PROCEDURE Atleastonedaypriortoinvestigation: 1. PutsomefertilesoilinaglassPetridishorclaycrucibleandheatuncoveredinan ovenatapproximately120Cforonehour. 2. Fillatightlysealablebottlewithbacterialgrowthmedia(about150mL)andadda smallamount(about1tsp)ofheatpretreatedsoil. 3. Growthebacterialcultureovernightat30C(orlongeratroomtemperature).The solutionshouldturncloudyandstarttofoam.Caution:Ifgrowinglongerthanone day,buildupofgaspressurecouldcausebottletorupture! Dayofinvestigation: 1. AssembletheMFCcellsaccordingtotheexplosionFigure3.8wewillcreatethree compartments(anode/cathode/anode).Separatethechamberswithionexchange membrane and use gaskets on both sides of the membrane to prevent leaking. Severalfuelcellsmaybejoinedtogethertogivegreatervoltage.

Figure3.8.The3chamberedMFC(topview)

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2. 3. 4. 5. 6.

Attachthetubingviatubeconnectorstotheinfluentport(side,bottom)oftheMFC chambers. Usingalligatorclipsandcopperwire,connectthetwoanodesinparallel(sothatthe twochambersbecomeelectricallyone). Connect the handheld multimeter in parallel to the MFC to investigate the development of cell potential over time. Attach one lead to the cathode and one leadtoeitheranode. Add 0.3 mL (300 uL) of methylene blue to each anode chamber through the top effluentport. Fillthetwo30mLsyringeswithcatholyteandanolytesolution(asfastaspossibleto limit aeration) and connect them to the respective tubings at the influent ports (Figure3.9).

Note:Toeliminatespacingissues,theinfluentportsforbothanodechambersshould beoneandthesameside,andtheinfluentportforthecathodechambershouldbe on the opposite side. Additionally, both anode chambers should be connected by tubingandaTshapedconnectorsotheycanbefilledsimultaneouslyfromthesame syringe.

7. 8. 9. 10.

Fill the cathode cell first (limits the exposure to oxygen in the anode chambers) followed by both anode cells (from the same syringe) taking care not to overfill them. Observe the cell potential on the multimeter and if it is significant (>450 mV), directlyconnecttheMFCtotheLEDinshortcircuit. The LED will flash for a short time until the potential drops below the required voltagenecessarytopowertheLED. DisconnecttheLED,waitforrecoveryofthecellpotential,andtryitagain.

Disposalandcleanup: TostoretheMFCsetupsandkeeptheelectrodesreusableandactive,itisnecessary to clean and disinfect the whole cell and especially the electrodes intensively with ethanolaftereachexperiment. Afteraddingasmallamountofbleachandashortincubation,spentbacterialbroth solutionscanbedraineddownthesink. Cathodicferricyanidesolutionishazardousandmayproducetoxicfumesincontact withstrongacids!Itmaybeusedseveraltimesandgetslightertocolorlessifthe electronacceptorisspent.Localregulationsshouldbeobservedwhendisposingof theusedsolution.

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 Figure3.9.Theassembled3chamberedmicrobialfuelcell

DISCUSSION The heat pretreatment of the soil killed all living cells and just the spores of bacteria strains survived.FermentativeClostridiumspeciesrepresentalargegroupofthesebacteriainsoil.If thesoilisinoculatedandincubatedanaerobicallyinarichbacterialmedium,thesporesofonly theanaerobicbacteriagerminateandformamixedbacterialculture.Astheygrowovernight, the fermentative Clostridium convert the glucose in the nutrient media mainly into butyrate, acetate, and molecular hydrogen. When this bacterial solution is introduced into the anode chamber,themethylenebluemediatorisreducedbythebacterialcellsaswellasbymolecular hydrogen. When the electrical circuit is closed, the methylene blue is backoxidized at the anodebydroppingitselectronsintotheelectricalcircuit.Theelectronsflowthroughthecircuit to the cathode chamber where the ferricyanide acts as the final electron acceptor. If the potentialdifferencebetweenthereducedanodesideandtheferricyanideonthecathodeside islargeenough,wecanpowertheLED.

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AnalternativeprotocolthatalsomayassistwithstudentsunderstandingofthefunctionalMFC would be to add methylene blue to the growing bacterial solution prior to the day of investigation,asopposedtoaddingitdirectlyintotheanodechamberoftheMFCduringthe actualinvestigation.Asthebacteriagrowandmetabolize,theywillreducethemethyleneblue toitscolorlessform.Whenthebacterialsolution(plusmethyleneblue)isthenintroducedinto theMFC,themethylenebluewouldbecomeblueagainwhenitisbackoxidizedattheanode. Thestudentswouldthenhaveavisualrepresentation(acolorchange)oftheelectronsbeing mediatedtotheanodebythemethyleneblue. FURTHERINVESTIGATIONS Iftimeallows,abacterialbiofilmcouldbedevelopedwithintheanodechambertocomparethe functionofthemethylenebluemediatedMFCtoamediatorlessMFC. Preparetheanodicfeedsolution o AnolytebacterialfeedsolutionforbiofilmMFC For1LofdiH2Oadd: - 0.8g sucrose - 3g KH2PO4 - 0.1g yeastextract - 0.033gNH4Cl - 0.06gK2SO4 - 0.033gFeCl2x4H2O - 0.011giron(III)citrate - 0.5gNaCl - 0.1gKCl - 0.1g CaCl2 - 0.1g MgCl2x6H2O AdjustpHto6.50.5andautoclave(alternatively,forschoollab,usefreshly boiled,cooleddownwaterforpreparationtomaintainsterilityuntiluse,do notletwaterstandopen.). Assemble the MFC kit as in the previous experiment, but in addition, seal the hole around the external anode connections (influent and effluent ports) to prevent the leakingofliquidandoxygenationoftheanodesolution.(Tip:Aquariumsiliconesealant workswell.)Also,tubingshouldbeconnected(andsealed)tothetop(effluent)portto allow the anode solution to be flushed out daily. To maintain anaerobic conditions in the anode chamber, be sure the tubing in the top (effluent) port remains closed (by clamp)exceptduringthetimeofsolutionreplacement.

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Mixverydifferentsamplesofanaerobicsoils,sludges,andwaterfromstagnantpuddles to get an anaerobic bacterial inoculum. Inject some of the inoculum into the anode chamberoftheMFC(throughinfluentport). Asinthepreviousexperiment,fillthetwosyringeswithanolyteandcatholytesolution, connect them to their respective tubings, and fill both cells with solution (through influentport). Addfreshanodesolutiononceperday.Byinitiallyconnecting(andsealing)tubingto thetop(effluent)portoftheanodechambers,(thespentsolutioncaneasilybeforced outasfreshsolutionisaddeddaily). ConnectthemultimeterinparalleltotheMFCasbeforeandperiodicallymeasurecell potential.Ifsignificantcellpotentialdevelops,connecttheMFCtoanexternalresistor (e.g., 100 Ohms, available at Radio Shack or similar electronics store). Wait until a stablepotentialisformedatthisresistance(couldtakedaysorweeks).Ifthepotential isstableovertime,theresistorcanbedisconnectedfor24hourstocollectpotential betweenthetwopoles.Ifthepotentialgoesover0.45V,theLEDdevicewillflash.Itis alsopossibletoperformapolarizationtestwithaseriesofdifferentexternalresistorsas itwasdescribedinChapter1,page37.

The anaerobic bacteria from the sludge mixture will grow in the anode compartment by utilizing the offered sucrose (sugar) in the feed solution. If the anodic chamber is kept air/oxygenfree,someofthebacteriawillbeginanaerobicrespirationbychoosingtheanodeas theterminalelectronacceptor(seeChapter2AnaerobicRespiration).Abiofilmwillthenbe slowlyestablishedattheanodethatdeliverselectronsintotheelectricalcircuit.
MFC protocol for school experiments was adapted from Bioscience Explained by D. Madden andJ.Schollar(http://www.bioscienceexplained.org/ENvol1_1/pdf/FulcelEN.pdf)

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Chapter4:SoNowWhat?MFC ChallengesandApplications

TECHNICALCHALLENGES Systemarchitecture.TheideaofusingbacteriainMFCstocaptureelectricityhasbeenaround forsometime,butinearlysystems,powerproductionwasverylowandrequiredtheaddition of extracellular mediators to shuttle electrons from inside to outside of the bacterial cell. In newersystems(e.g.,mediatorlessMFCsthatemploybiofilms);mediatorsarenotneeded,and power production from MFCs has increased dramatically in just the past few years, in part becauseofdesignsthatlowerinternalresistance(seeChapter3MediatorlessMFCs). Resistance can be described as anything that opposes the flow of electric current. Often the system components of the MFC themselves are regarded as the principle (internal) resistors (e.g., transfer resistance between electron mediator and anode or electrolyte and cathode). Forexample,justa0.1Ohmresistanceoccurringatacurrentof2ampscanyielda0.2Vdropin theelectricalpotentialoftheMFC(seeChapter1ElectricalCircuits).Thetheoreticalpower (orEMF)generatedbyMFCsisafunctionoftheindividualelectrodepotentialswhenthecircuit is open, but the actual power generation of the MFC is often less than what is theoretically possible. Theanodepotentialissetbytherespiratoryenzymesofthebacteriaanddoesnotappearto varysubstantiallyindifferentsystemsorwithdifferentsubstrates(fuels).Ontheotherhand, the cathode potential varies depending on the catholyte and oxidant. Experimentally, lower thanexpectedvoltagesareusuallyobtainedwhenoxygenisusedasthefinalelectronacceptor. Withferricyanideasthefinalelectronacceptorinsteadofoxygen,lessinternalresistanceand higher cell voltages have been documented, however, power generation with ferricyanide is notsustainabletheferricyanidemustbeexternallyregeneratedovertime. Withoxygenastheelectronacceptoratthecathode,themaintechnicalchallengeinimproving power generation is to create a system architecture that minimizes the internal resistance of theMFCbut,atthesametime,allowsforcontinuousflowthroughthesystem.Onepossible remedy could be the use of an air cathode. Exposing the cathode directly to air (an air cathode),insteadoftodissolvedair(oxygen)inwater,hasresultedinsubstantialincreasesin system performance (Figure 4.1). Clearly, though, maximizing power generation in MFCs will require innovative flow patterns and increased electrode electrolyte interactions that minimize internal resistance. Additionally, finding methods to increase the cathode potential, withoxygenastheelectronacceptor,couldhaveasubstantialimpactonpowergeneration. Materials. The cost of materials used to construct MFCs will also be a key factor for the successfulapplicationofthetechnologyonalargerscale.Verylargesurfaceareasareneeded

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for supporting bacterial biofilms, and the structure must be able to bear the weight of the waterandbiofilm.Electrodematerialsrangefromcarbonclothandcarbonpaper,tographite rods,plates,andgranules.Cathodesaremadefromthesamematerials,buttheyalsocontain preciousmetals,suchasplatinum,whenoxygenisusedastheelectronacceptor. Somematerialsarenotexpectedtobesuitableforscaleupbecauseoftheirinherentlackof durability or structural strength (e.g., carbon paper), or cost (e.g., platinum catalysts, ion exchange membranes). Future designers will need to consider conductive coatings on structurallystrongsupportingmaterials.Systemscaleupwillalsorequirethatthedesignand applicationofthesematerialsbeadaptabletomassmanufacturingapproaches. Microbiology.Ourunderstandingofelectrochemicallyactivemicrobesisstillinitsinfancy,but clearlyawholenewfieldofmicrobialecologyisemergingthatisbasedonanodophilicbacteria (bacteria that will adhere to an anode) and possible interspecies electron transfer. These bacteria may be referred to as electricigens based on their ability to exocellularly release electrons. The initial understanding of electron transfer by bacteria to electrodes came from studies of metalreducing bacteria, such as Geobacter and Shewanella species, which can produceelectricityinMFCs.Biochemicalandgeneticcharacterizationsindicatedthatforsome bacterial species, outermembrane cytochromes (membranebound proteins containing iron thatcarryoutelectrontransport)canbeinvolvedinextracellularelectrontransfer.Also,some bacteria produce and use soluble electron shuttles that eliminate the need for direct contact betweenthebacterialcellandtheelectronacceptor(Figure4.1). The recent discovery of nanowires introduces a whole new dimension to the study of extracellular electron transfer. These conductive, pillilike structures identified, thus far, in certainGeobacterspeciesappeartobedirectlyinvolvedinextracellularelectrontransfer(see Chapter 3 Mediatorless MFCs). These nanowire structures allow the direct reduction of a distantelectronacceptor.Thisremovestheneedforsolublemediatorsthatwouldbelostina continuousflowMFCandmayallowfordirectinterspecieselectrontransfers.

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Figure4.1.Summaryofmethodsforelectrontransfertotheanode
(withpermissionfromtheNaturePublishingGroup,License#2342510598123)

Glucose serves as an example fuel. a | An indirect microbial fuel cell. A fermentative microorganism converts glucose to an end product, hydrogen, which can react with the anode to produce electrons and protons. This process only partially recovers the electrons available in the organic fuel as electricity, and results in the accumulation of organic products in the anode chamber. b | A mediatordriven microbial fuel cell. An electron shuttlingmediatoracceptselectronsfromreducedcellconstituentsandtransferstheelectronstotheanode.The reoxidizedmediatorcanthenundergorepeatedcyclesofreductionandoxidation.Inmostinstances,thecellsthat have been used in such fuel cells only incompletely oxidize their organic fuels as shown. c | The oxidation of glucosetocarbondioxidewithdirectelectrontransfertotheelectrodesurface.Glucoseistakenintothecelland oxidizedtocarbondioxidebytypicalmetabolicpathways.Electronsderivedfromglucoseoxidationaretransferred across the inner membrane and outer membrane through electron transport proteins, such as ironcontaining cytochromes. In this example, the system is illustrated with an air cathode rather than a cathode submerged in water.d|Atwochamberedmicrobialfuelcell.Thissystemisnotoptimizedformaximumpowerproductionbutis convenientformicrobiologicalstudies.

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Activity4.1Electricigens:MicrobialEnergizers Studentsshouldreadthearticlefoundat: http://www.asm.org/microbe/index.asp?bid=43711 After reading, have students answer the questions that follow. This reading assignment will also provide some background information on current MFC applications and technologythatwillassistwiththeposterproject.

Whatareelectricigens? Microorganismswiththeabilitytooxidizeorganiccompoundstocarbondioxide whiletransferringelectronstoelectrodeswithhighefficency. How are the actions of electricigens similar to the functioning of a traditional hydrogen fuelcell? Botharecapableofconvertingafuelintoelectricitywithoutlosingsubstantialamounts ofenergyasheat.Ahydrogenfuelcelloxidizeshydrogenandreducesoxygentowater whileproducingelectricity.Electricigensutilizetheirowncellularmetabolicpathwaysto oxidizesugarsandreduceoxygen(oranotherfinalelectronacceptor). What are the differences between a hydrogen fuel cell and MFCs that employ electricigens? Hydrogenfuelcellsrequireaverypuresourceofahighlyexplosivegasthatisdifficultto store and distribute, and hydrogen is derived mainly from fossil fuel rather than renewable sources. In contrast, the energy sources for microbial fuel cells that utilize electricigens are renewable organics, which are stable and very cheap. In terms of powerproduction,MFCswillneverbeabletocompete(intermsofelectricityproduction) withthetraditionalfuelcellbecausethebiomassfueloftheMFCwillalwaysbeinlower concentrations than what is fed into a hydrogen fuel cell. Thus, the power capable of beinggeneratedbyMFCswillalwaysbeordersofmagnitudelowerthanhydrogenfuel cells.

Activity4.2MFCApplicationsPoster IndividualstudentsorstudentgroupswillresearchthevariousapplicationsofMFC technologycurrentlyinuseandthenpresenttheirfindingsinapostersession.

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Activity4.2MFCApplicationsPoster
MATERIALS Internet/collegelibraryaccess,posterboard,markers,coloredpencils PROCEDURE IndividuallyorincooperativegroupsthestudentsshouldresearchtheapplicationsofMFC(or BES)technologythatarecurrentlyinuseorthathavebeenproposed.Havestudentsperform some initial research into possible applications, or alternatively, present the information on applicationsthatfollowthisactivityduringclasslecture.Then,eachstudent(orgroup)should select their topic. After a predetermined period of time, the students should present their findingstotheclass. Havethestudentsfocustheirresearchtoansweringthesequestions: WhatisthespecificapplicationoftheMFC,orwhatdoestheMFCpowerorproduce? Where(location)isthistechnologyinuse? Howlonghasitbeenusedandhowsuccessfulistheventure? BasedonourstudyoftheMFC,describehowtheMFCisbeingusedinthistechnology? Forexample: o Isitmediatorlessorisamediatorused? o WhatreactionsareoccurringattheanodeandcathodeoftheMFC?

Asuggestedrubricforgradingtheposterisprovidedattheendofthischapter.

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WASTEWATERANDOTHERAPPLICATIONS Whenanewtechnologyisintroducedintothemarketplace,thegreatestlikelihoodforsuccess occurs when the most immediately profitable application is targeted first. As the technology develops, becomes better understood, and improves, more difficult applications can be undertaken.ThemostimmediateandusefulapplicationsofMFCsappeartobeforwastewater treatmentandother,similarnicheapplications.Renewableenergyproductionisalongerterm prospect that will require substantial technical and manufacturing advances. But research advances during the last three years also showed promising other applications, besides microbiallyinducedpowergeneration,asinthecaseofanMFC.Tonameallnewtechnologies thatarebasedonthesamefunctionalprinciples,butcanbeusedfordifferentapplications,the termBioelectrochemicalSystem(BES)isgettingpromoted. Wastewater treatment. Worldwide, more than 2 billion people do not have adequate sanitation,inlargepartbecauseofalackofstartupcapitalaswellasoperatingcosts.Inthe U.S.,approximately$25billionisspentannuallyforwaterandwastewatertreatment.Overthe next 20 years, water and wastewater infrastructure demands will require over $2 trillion for building, maintaining, and operating these systems. In the U.S., close to 4% of the total electricity produced is used for the operation of the whole water and wastewater infrastructure.AtreatmentsystembasedonanMFCprovidesagreatopportunitytodevelop thetechnologybecausethesubstrateisfreeandwastewatermustbetreated. Current methods for wastewater treatment utilize a filtration/straining procedure to remove large objects from primary wastewater and then employ an aerobic treatment of secondary wastewater to degrade the biological content of the sewage. Large amounts of oxygen are pumped into secondary wastewater, at an immense economical and electrical cost, creating what the industry refers to as activated sludge. Essentially, bacteria undergo aerobic respirationandmetabolizethewastecomponentsofthesewage,usingitasafoodsource. MFCscouldbeusedinatreatmentsystemasareplacementfortheexistingenergydemanding aerobictreatment;however,wedonotyetknowhowtoeconomicallyscaleupanMFCorwhat thecostswouldbetoreplaceaconventionalsystemwithanMFCbaseddesign.Scaleupand materials issues are the greatest challenges in the application of MFCs for wastewater treatment,butpotentially,energyrecoveryatwastewatertreatmentplantscouldleadnotonly to a sustainable system based on energy requirements (an energy neutral wastewater treatmentfacility),butpossiblytotheproductionofanetexcessofenergy. Environmentalsensors.Dataonthenaturalenvironmentcanbehelpfulinunderstandingand modeling ecosystem responses, but sensors distributed in the natural environment require powerforoperation.MFCscanpossiblybeusedtopowersuchdevices,particularlyinriverand deepwater environments where it is difficult to routinely access the system to replace batteries.Sedimentfuelcellsarebeingdevelopedtomonitorenvironmentalsystems,suchas creeks,rivers,andoceans.Powerforthesedevicescanbeprovidedbyorganicmatterinthe sediments. Power densities are low in sediment fuel cells because of both the low organic 86

matter concentrations and their high intrinsic internal resistance; however, the low power densitycanbeoffsetbyenergystoragesystemsthatreleasedatainburststocentralsensors (Figure4.2).

Figure4.2.Asedimentmicrobialfuelcellforuseasanenvironmentalsensor
(withpermissionfromtheNaturePublishingGroup,License#2342510598123)

a|Aschematicofasedimentmicrobialfuelcell.OrganismsinthefamilyGeobactercanoxidizeacetateandother fermentationproducts,andtransfertheelectronstographiteelectrodesinthesediment.Theseelectronsflowto thecathodeintheoverlyingaerobicwaterwheretheyreactwithoxygen.b|Anactualsedimentfuelcellbefore deployment.

Bioremediation.AnMFCcanbemodifiedininterestingandusefulways,andthiscanleadto new types of fuel cellbased technologies. With such modifications, however, these systems maynolongerbetruefuelcellsbecausetheydonotproduceelectricity.Onesuchapplicationis themodificationofthebasictwoelectrodesystemforbioremediation.TheMFCisnotusedto produce electricity; instead, power can be put into the system to drive desired reactions to removeordegradedangerousortoxicchemicalstosaferorusefulforms. Renewable electricity production from biomass. Because of uncertainty about the materials neededandtheircosts,combinedwithcomparativelylowcostsforoil,theapplicationofMFCs forrenewableenergyproductionfromcrops,suchascorn,isnotlikelyintheimmediatefuture. In the near term, MFCs will have to compete with more mature renewableenergy technologies, such as wind and solar power. The operating costs needed for electricity productionwithMFCswillprobablybetoogreatifthesubstratefortheMFCisgrownasacrop inamannersimilartothatforethanolproductionfromcorn.

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Microbialelectrolysiscells(MEC).Inadditiontogeneratingelectricalenergy,MFCscouldalso beusedinaBEStogenerateindustrialchemicalsfromwasteproductswithexternalinputof smallamountsofenergy.MECsareveryversatilesystemsinwhichatleastoneoftheanodicor cathodic oxidationreduction reactions are microbially catalyzed. They are capable of generating products , such as hydrogen, methane, and ethanol. Likewise, they have the potential to catalytically degrade less desired reactants and contaminants into more economicallyprofitableorecologicallysoundproducts. BESshavealsobeenusedsuccessfullytoremovesulfurcomponents,promotedenitrification, andreduceperchloratesandchlorinatedorganiccompounds. EDUCATION Some of the most immediate and useful applications of MFCs are in the classroom: Students findelectricitygenerationbybacteriabothfascinatingandfun!MFCshavebeenfoundtobean effectiveeducationaltooltocapturestudentinterest.Thesesmallandportablesystemsserve asawonderfulplatformformotivatingstudentstostudyandunderstandcomplexconceptsof cell respiration, microbial ecology, electrochemistry, and materials science. Additionally, processes that can couple sustainable energy production with waste treatment have innate appealtoenvironmentallymindedstudents.TheMFCisamodelsystemforscienceinstruction that dissolves disciplinary boundaries and shows how technology can help solve significant socialandenvironmentalissues.Directevidencefortheappealofthistechnologycanbeseen throughrecentsciencefairprojectsonMFCsbystudentsinmiddleandhighschoolsaroundthe world. At university level, the MFC has been used in undergraduate and graduate laboratory courses in environmental microbiology to teach students methods of microbialcommunity analysis.

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RubricforMFCApplicationsPoster

Criteria Organization

1
Cluttered,nodefinitive sections,allovertheplace

2
Noheadings,butsectioned Hardtofollow,requires assistance Obviousrefinement required Verylittleuseofcoloror picturesbutenoughto engageandholdattention

3
Headingspresentbut unclear Mustrereadforclarity Someevidenceof refinement Someuseofcolor, diagrams,etc. Willengagebutwillnot stimulate

Creativity

Contentand Literacy

Bland,novariability Nouseofcolorordiagrams Boringtolookat,doesnot catchyourattention Interest,motivation,effort andtimeobviouslyabsent Noanalysisoftopic Noexplanations Nospecificconnectionsto MFCunit Nouseofresources

Poorexplanations Inaccurateconnectionsto MFCunit Misinterpretsthe technology Oneresourceforsure

Difficulty(depthofthought) Leveland notsuitableforgrade Difficultyof level/notrelatedtoclass Understanding discussions(tooeasy)

Explanationdescribes minimallevelofvalidity Needsseriousrefinement

(Depthof Thought)

Definedsections Clearheadings Flowsnicelytoassistthe readerwithouthelp Finishedproduct Interesting,engaging, visuallystimulating Aestheticallyappealinguse ofcolor,diagramsandtext Interest,motivation,effort andtimeobviouslypresent Adequateexplanations Conceptfullyandproperly MFCunitconnections explained presentbutcouldbe Insightpresent developedfurther MFCunitspecific Morethanoneresource connectionsmade present Contentisaccurate, comprehensiveandwell supported Excellentuseofresources Difficultycouldbeincreased Difficultyappropriatefor ordeveloped gradelevel Somelevelof Understandingpresentand understandingshown apparent

Superficial/irrelevant