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The Value of Inoculum Management in Microalgae Production Valerie Harmon Harmon Consulting, Inc. Co-Authors: J. McGowen, P. Pienkos, T. Lane, K. Poorey, E. Knoshaug, E. Wolfrum

The Value of Inoculum Management in Microalgae Productionalgaebiomass.org/wp-content/gallery/2012-algae-biomass-summit/20… · Chris Withstandley Mary Riddle Nancy Pham Ho (FIT)

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Page 1: The Value of Inoculum Management in Microalgae Productionalgaebiomass.org/wp-content/gallery/2012-algae-biomass-summit/20… · Chris Withstandley Mary Riddle Nancy Pham Ho (FIT)

TheValueofInoculumManagementinMicroalgae

ProductionValerieHarmon

HarmonConsulting,Inc.Co-Authors:J.McGowen,P.Pienkos,T.Lane,K.Poorey,E.Knoshaug,E.

Wolfrum

Page 2: The Value of Inoculum Management in Microalgae Productionalgaebiomass.org/wp-content/gallery/2012-algae-biomass-summit/20… · Chris Withstandley Mary Riddle Nancy Pham Ho (FIT)

• Cultureswillinitiateinalaboratoryinsmallvolumesandarethentransferredincrementallyuntilfullscaleisachieved.• Typicalscaleupwouldassume10– 20%inoculumbyvolumeasthecultureistransferredfromonescaletoanother.• Initialscaleupistypicallyabatchproductionmodewhichmeansthealgaeismovedoutofonesystemcompletelyintothenextculturestage• Asscaleincreases,inoculumsystemscanmovetobatch,semi-continuousorcontinuouscultivationsystems

HarmonConsulting,Inc.

Necessaryforachievingscale Anexampleofaproductionschemeforbatchcultureofalgae(LeeandTamaru,1993).

Inoculum:Whyisitimportant?

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Whathappenswhenscaleisachieved?

• Isinoculumnecessaryformaintainingscale?• Algaecultivationsystemsdofail!Thisistrueofpondsandofclosedsystems!

• Wearejustbeginningourstudyofculturesystemfailuresasanindustry,whichwillenableustocontrolandmanageoursystemsforimprovedreliabilityandproductivity.

• Howwillsystemrecoverybemanagedinatimeandcosteffectivemannerwhenlargescaleproductionsystemsfail?• Thereisnotjustonewaytoapproachinoculumgeneration

• Batchproduction• SemiContinuousproduction• Continuousproduction

HarmonConsulting,Inc.

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HarmonConsulting,Inc.

Anexampleofalgalgrowthdynamics

AlgalGrowthDynamicsDuringScaleUp(FAOFisheriesTechnicalPaper361,1996).EditedbyLavens andSorgeloos

Lag“phase,duringwhichlittleincreaseincelldensityoccurs,isrelativelylongwhenanalgalcultureistransferredfromaplatetoliquidculture.Culturesinoculatedwithexponentiallygrowingalgaehaveshortlagphases,whichcanseriouslyreducethetimerequiredforupscaling.Thelagingrowthisattributedtothephysiologicaladaptationofthecellmetabolismtogrowth,suchastheincreaseofthelevelsofenzymesandmetabolitesinvolvedincelldivisionandcarbonfixation.”

“Inpractice,culturecrashescanbecausedbyavarietyofreasons,includingthedepletionofanutrient,oxygendeficiency,overheating,pHdisturbance,orcontamination.Thekeytothesuccessofalgalproductionismaintainingallculturesintheexponentialphaseofgrowth.”

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“Themostimportantfactorsgoverningcontaminantgrowthdynamicsinalgalmassculturesarethegrowthrateofthecontaminant,therateofdilutionofthebioreactorcontainingtheculture,andthesizeoftheinitialinoculum”ofthecontaminant

HarmonConsulting,Inc.

ModeledContaminantGrowthDynamics Conclusion

ForeheadandO’Kelly,2013SmallDoses,BigTroubles:ModelingGrowthDynamicsofOrganismsAffectingMicroalgalProductionCulturesinClosedPBRs

Page 6: The Value of Inoculum Management in Microalgae Productionalgaebiomass.org/wp-content/gallery/2012-algae-biomass-summit/20… · Chris Withstandley Mary Riddle Nancy Pham Ho (FIT)

RealLifeExamplesATP3TestBedDataonPondReliabilityandPersonalExperience

HarmonConsulting,Inc.

Page 7: The Value of Inoculum Management in Microalgae Productionalgaebiomass.org/wp-content/gallery/2012-algae-biomass-summit/20… · Chris Withstandley Mary Riddle Nancy Pham Ho (FIT)

AlgaeTestbedPublicPrivatePartnership

HarmonConsulting,Inc.

EstablishCollaborativeOpenTestbedsatfivegeographicallydistinctproductionfacilitiesinordertoquantifyseasonaland

environmentalvariabilityinalgaeproduction

• SoutheastCoastal:FL• WesternCoastal:CA• PacificTropicalCoastal:HI

• SoutheastInland:GA• SouthwestDesertInland:AZ

Page 8: The Value of Inoculum Management in Microalgae Productionalgaebiomass.org/wp-content/gallery/2012-algae-biomass-summit/20… · Chris Withstandley Mary Riddle Nancy Pham Ho (FIT)

AlgaeTestbedPublicPrivatePartnership

GenerateHigh-ImpactDatafromLongTermCultivationTrials

HarmonConsulting,Inc.

• UnifiedFieldStudies(UFS)• All5sitesexecutingsameproductionmethods,systems,includingseedproductionsystems,andstrains

• ScientificDataManagementSystemandHarmonizedSOPsforanalyticalandUFSproductionmethods• Dataintegrity• StrongQCcapabilityofdata

• DatapubliclyavailableviaNationalRenewableEnergyLabwebsite

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UnifiedFieldStudyExperimentalDesign

• FiveSites:Southwestdesertinland, Southwestcoastal,Southeastcoastal,Southeastinland,andPacifictropicalcoastal.• FourSeasons:Spring (March– May),Summer (June–August),Fall (September– November),Winter(December– February)• TwoYears: 2014and2015• ThreeStrains:Nannochloropsisoceanica (CellanastrainKA32),Chlorellavulgaris(ASUstrainLRB_AZ_1201),Desmodesmuscf intermedius(CellanastrainC046)• ExperimentalTreatments:Harveststrategies:frequency(Onceperweek(1x)vsThreetimesperweek(3x)) and dilutionrates(0.11,0.214and0.3perday)

HarmonConsulting,Inc.

Page 10: The Value of Inoculum Management in Microalgae Productionalgaebiomass.org/wp-content/gallery/2012-algae-biomass-summit/20… · Chris Withstandley Mary Riddle Nancy Pham Ho (FIT)

UFSInoculumProductionMethodsforSystemInitiation

HarmonConsulting,Inc.

Initialculturedensity~0.1-0.2g/LAveragegrowthrate~0.3g/L/dayGrowthperiodis9-10daysFinalculturedensity~3.5g/L

1to1transfer

Columntopanel

Initialculturedensity~0.2g/LAveragegrowthrate~0.15-0.2g/L/dayGrowthperiodis9-10daysFinalculturedensity~2.0g/L

12-2’x2’panelsyieldprovidebiomassforaminimumtargetof0.05g/L(AFDW)initialmini-pondconcentration(siteshave14available)

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UFSInoculumforSystemRecoveryPost-FailureHealthyPond

FailedPondL

SameTreatment

Drain,clean,calibrateAdvantageofnogrowout,rightbacktoharvestingimmediately!Butisthisqualityinoculum?

HarmonConsulting,Inc.

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PondReliabilityMetrics

• Reliability:Theabilityofanasset(inthiscaseapond)toperformitsintendedfunctionwithoutfailureforaspecifiedperiodoftimeunderspecifiedconditions.• Metrics• MeanTimeBetweenFailure(MTBF)inDays

• Aharmonizedmetricasperthe SocietyforMaintenanceandReliabilityProfessionals• Averagelengthofoperatingtimebetweenfailures;error-freeperformancetime• Thismetriccanonlybecalculatedifapondfails

• MeanTimeToReset(MTTR)inDays,2.26days• Annual#ofPondResets

• 365/(MTBF+MTTR)

• AnnualResetEffortinDays• Annual#ofresetsxMTTR

HarmonConsulting,Inc.

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HarmonConsulting,Inc.

MeanTimeBetweenFailures Effort

BatchSeedInoculumvsSemi-ContinuousOpenPondInoculum

19.98

12.79

0

5

10

15

20

25

Seed Pond

Days

InoculumSource:MeanTimeBetweenFailures

21.00

38.5847.37

87.16

0

20

40

60

80

100

120

Seed Pond

Days

InoculumSource:AnnualResetEffort

Annual#ofRepairsAverageAnnualRepairEffort(#Repairs*MTTR)Average

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ReliabilityImprovementsRelatedtoInoculumandProductionStrategies– ANumbersGame• SeedInoculumassumedtostartwith0ciliatesandprotozoaperml,butonday3itisassumedthat5ciliatesand1protozoaentertheculture

• PondInoculumassumedtostartwith150ciliatespermland50protozoapermlonday0(pondinoculum)• Ciliatepestsassumedtoincreasetheirpopulationsbyafactorof4daily• Otherprotozoanpestsassumedtoincreasetheirpopulationsbyafactorof2.67daily

0

5E+10

1E+11

1.5E+11

2E+11

2.5E+11

0 2 4 6 8 10 12 14 16

#/ml

Days

TheoreticalContaminantGrowthRatesSeedInoculum:VariousHarvestStrategies

Ciliates:3x,0.214 Protozoa:3x,0.214

Ciliates:1x,0.11 Protozoa:1x,0.11

0

2E+11

4E+11

6E+11

8E+11

1E+12

1.2E+12

1.4E+12

1.6E+12

1.8E+12

0 2 4 6 8 10 12 14 16

#/ml

Days

TheoreticalContaminantGrowthRatesPondInoculum:VariousHarvestStrategies

Ciliates:3x,0.214 Protozoa:3x,0.214

Ciliates:1x,0.11 Protozoa:1x,0.11

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BatchInoculumProductioninOpenPonds:LifeLessonsfromAustralia

• Noinoculuminitially,restartedfailedpondsfromanexistingponds• Notachievingbiomassproductiongoals

• Scalefromoneculturesystemtothenext• Lab– tominiponds(700Lvolume)

• Labtooutdoortransition• (14)minipondsto(4)50m2 ponds• (4)50m2 pondsto(2)400m2 ponds• (2)400m2 pondsto(1)4000m2 ponds

• Inoculumnotneeded– toharvestbasinandbiomassnotlost!!!!!• Wheninoculumwasneededonepondperweekcouldbeinoculated• Thisinoculumsystemutilizedexistingcapabilityandstaffing,socostnothingtoimplementotherthannutrientswhichproducedeitherinoculumorharvestablebiomass• Wellworthit!Producedqualityinoculum!• Existingstaffhadaroutinethatprovidedtheinoculumweneededwhenweneededit!• Weachievedourbiomassproductiongoals!

HarmonConsulting,Inc.

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Conclusions:InoculumBestPractices

• Aretherewaystomanageourinoculumthatwillhaveasignificanteffectonmicroalgaeproduction?• Quality

• Freeofcontamination,oratleastascleanaspossible

• Fastgrowing,notpaststage2• Nutrientreplete

• Providelittletonotransferstress• Qualityoftheinoculumwilldeterminethelongevityoftheproductionsystem• Testbeddatacanprovidedirectionintermsofhowbesttomanageourinoculumforlargescaleproduction• Inoculumresearchison-goingincurrentATP3research

HarmonConsulting,Inc.

Page 17: The Value of Inoculum Management in Microalgae Productionalgaebiomass.org/wp-content/gallery/2012-algae-biomass-summit/20… · Chris Withstandley Mary Riddle Nancy Pham Ho (FIT)

HarmonConsulting,Inc. 17

Acknowledgements

FloridaAlgaeStevenSchlosserChrisWithstandleyMaryRiddleNancyPhamHo(FIT)

CellanaMartinSabarskyJohannaAntonMarcelaSaraccoDavidAntonKeao Bishop-Yuan

Valicor RenewablesKiranKadamBrianGoodall

ASUGaryDirksJohnMcGowenThomasDempsterPeteLammersMiltSommerfeldWilliamBrandtJessicaChengSarahArrowsmithSarahKempkesDavidCardelloTheresaRosovMariaBautistaMaryCuevasRichardMalloyHenriGerkenPierreWenselLindaBoedekerJamieRockSarahMasonTravisJohnsonSydneyLines

UTEXSchonnaManningJerryBrand

NRELPhilPienkosLieveLaurensEricKnoshaugEdWolfrumRyanDavisChristopherKinchinStefanieVanWychenSandiaNationalLabsRonPateToddLaneKunalPooreyPatriciaGharagozlooThomasReichardtJeriTimlinJessicaDrewryPamelaLane

CalPolyTrygLundquistBradenCroweEricNicolai

CommercialAlgaeProfessionalsAlbertVitaleJeremyWeir

ASUUndergradsShaylinMcgheeDanielBreyfogleLeviBunkerThomasChengattuDelaneyDeHertoghMark FordRileyGreerDilon HurleyKevinLawsonShelbyLuiCarlosLunaMiguelMartinezJoePalaskiCaseyPetersonJeremyPoetAlexandraPrassasSherrySaethao

JayTenisonJessicaVaughnWyatt WesternMykalYbarra

GTUndergradsFariha HassanJerryDuncanFrazierWoodruffShusuke DoiHao FuPatriciaPenalver-ArguesoAllison DunbarAllisonCarrSichoonParkPriyaPradeepTerrySnell

CatherineAchukwuChristineYi

CPUndergradsAydee MelgarGulce OzturkKaitlynJonesMichaelAntoineTrung KTranJakeBenderHeatherFreedDanielMcBroomMicheleHendricksonGerardNguyenDevenDilibertoJackSunderlandDanAverbujAnnMarieSequeiraLaurenMiller

MicheleHendricksonEmilyWangJackSunderlandAnnMarieSequeiraSoroush AboutalebiLaurenMillerSamanthaLuiMicheleHendricksonGabriellaCamposWillBrilesLettyThottathil

GeorgiaTechYongshengChenStevenVanGinkelThomasIgouYingiang SunZixuanHu

HarmonConsultingInc.ValerieHarmon

Page 18: The Value of Inoculum Management in Microalgae Productionalgaebiomass.org/wp-content/gallery/2012-algae-biomass-summit/20… · Chris Withstandley Mary Riddle Nancy Pham Ho (FIT)

Thankyouforyourtimeandattention!