14
Bigelo R T ow Lab Research The Gulf REU borato Experien of Main U Symposium Thursday 1 ory for nce for U ne and the m Program y, August 9, Ocean Undergra e World & Abstract , 2012 n Scien aduates Ocean ts nces

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Page 1: Bigelow Laborato ry for Ocean Sciences · PDF fileassimilation iatom Thala and one bac xleyi had lo ... ipid product tments betw d maximum controls. Str ... leting nitrat BODIPY 50

Bigelo

RT

ow Lab

Research The Gulf

REU

borato

Experienf of Main

U SymposiumThursday

1

ory for

nce for Une and the

m Program y, August 9,

Ocean

Undergrae World

& Abstract, 2012

n Scien

aduates Ocean

ts

nces

Page 2: Bigelow Laborato ry for Ocean Sciences · PDF fileassimilation iatom Thala and one bac xleyi had lo ... ipid product tments betw d maximum controls. Str ... leting nitrat BODIPY 50

2

Program 1:00 Opening Remarks 1:15 Marie Neidig- University of Southern Maine

PRODUCTION AND ASSIMILATION OF DIMETHYLSULFONIONPROPIONATE (DMSP) BY MARINE PHYTOPLANKTON (Matrai – Archer)

1:30 James (Nick) Hunt- Texas A&M University – Corpus Christi.

ENHANCEMENT OF LIPID PRODUCTION IN MICROALGAE BY DODECANE ADDITION AND NITRATE LIMITATION (Wilson- Countway)

1:45 Cynthia Liles- University of California, Riverside

THE RELATIONSHIP BETWEEN MICROALGAE GROWTH PHASE AND LIPID CONTENT (Countway-Wison)

2:00 Emily Bell-Hoerth- Earlham College

VARIATIONS IN PHYTOPLANKTON COMMUNITIES ASSOCIATED WITH BIVALVE AQUACULTURE IN COASTAL MAINE (Poulton - Heil)

2:15 Zakary Jaques- Colby College

EFFECTS OF DISSOLVED HUMIC COMPOUNDS FROM THE DAMARISCOTTA AND KENNEBEC RIVERS ON MARINE MICROBIAL COMMUNITIES. (Poulton - Heil)

2:30 Andrew Burchill – University of Chicago

DETECTING EVOLUTIONARY SELECTION IN HIGHLY CONSERVED PROTEINS: PHYSIOCHEMICAL SHIFTS IN THE MAMMALIAN CYTOCHROME B (McClellan)

2:45 Break (15 minutes) 3:00 Ashley Poehls – Lake Superior State University

EFFECTS OF OCEAN ACIDIFICATION ON GROWTH RATE, BIOMASS, AND COCCOLITH MORPHOLOGY OF THE COCCOLITHOPHORE PLEUROCHRYSIS (Balch - Fields)

3:15 Melanie Ross - Colby College

INDIRECT EFFECTS OF OCEAN ACIDIFICATION ON ACARTIA TONSA (Balch - Fields)

3:30 Alyson Lowell - University of Maine

EXPERIMENTAL TECHNIQUES FOR CHARACTERIZING SHEATH-FORMING ZETAPROTEOBACTERIA: A METHODOLOGICAL APPROACH (Field-Emerson)

3:45 Kaitlyn Pritchard – Northeastern University

EXAMINING THE GRAZING PREFERENCES AND DIGESTIVE PROCESSES OF OXYRRHIS MARINA USING A RATIOMETRIC FLUORESCENT DYE (Neuster-Twinning)

Page 3: Bigelow Laborato ry for Ocean Sciences · PDF fileassimilation iatom Thala and one bac xleyi had lo ... ipid product tments betw d maximum controls. Str ... leting nitrat BODIPY 50

3

Abstracts

Page 4: Bigelow Laborato ry for Ocean Sciences · PDF fileassimilation iatom Thala and one bac xleyi had lo ... ipid product tments betw d maximum controls. Str ... leting nitrat BODIPY 50

PROMAR Mari1Uni DimantiosulfupartiDunatertiorespeno adhuxleDMSloweabunDMS

ODUCTIONRINE PHY

ie Neidig1,2, iversity of So

methylsulfonioxidant and/our for bacteriiculate DMSaliella tertioolecta produectively. Totdjustment toeyi. T. pseudSPd did not est in bacterindance and aSPd levels.

N AND ASSTOPLANK

Dr. Patriciaouthern Mai

opropionateor cryoproteial organism

SP(p) by threolecta and a uced no DMStal DMSPp c

o intracelluladonana enhachange as a ized treatmenactivity in th

IMILATIOKTON

a Matrai2 & Dine, 2 Bigelow

(DMSP) is ectant. DMSP

ms. We measuee axenic phycoccolithophSPp, while Tconcentratioar DMSP witanced its DMfunction of nts, especiale saline cond

ON OF DIM

Dr. Steve Arw Laborator

produced byP, when releured the proytoplankton hore EmilianT. pseudonanons for all spth cell growt

MSPp producsalinity, aftelly at 50 g/kgdition, despi

4

METHYLSU

rcher2 ry for Ocean

y marine phyeased into seduction and species (a d

nia huxleyi) na and E. huecies increasth. DMSPd wction under ler the initial g salinity. Thite decreased

ULFONION

n Sciences

ytoplankton eawater, is an

assimilationdiatom Thala

and one bacuxleyi had losed as a funcwas assimilalow salinity cosmotic shohis is likely d phytoplank

NPROPIONA

worldwide an important n of dissolveassiosira tertcterized straiow and high ction of cell ated only by conditions w

ock. DMSPddue to the hikton abundan

ATE (DMS

as an osmolysource of ca

ed DMSP(d)tiolecta, a chin of T. pseuDMSPp probiovolume dT. pseudona

whereas the rd concentratiigher bacterince and unch

P) BY

yte, arbon and and hlorophyte

udonana. D. oduction, displaying ana and E. release of ions were ial hanged

Page 5: Bigelow Laborato ry for Ocean Sciences · PDF fileassimilation iatom Thala and one bac xleyi had lo ... ipid product tments betw d maximum controls. Str ... leting nitrat BODIPY 50

ENHNIT

AlgaphotThisPavlLipidwerecultucytomfactoan inbetwprod

HANCEMERATE LIM

1T

ae, a renewabtosynthetic o research foclova lutheri cd concentrate divided intures. Samplemetry. The mor 15 for the ncrease by th

ween days 6 aduction, and

ENT OF LIPMITATION

J. NTexas A&M

ble fuel sourorganisms prcused on enhcultures withtions were mo two treatm

es were stainmain findingP. lutheri do

he average faand 13 compare a solid fo

PID PRODU

Nicholas HuUniversity –

rce, can poteroduce lipidshancing lipidh the hydroc

monitored usiments: dodecned with a 1:gs display a lodecane treaactor 2.52 anpared to the foundation fo

UCTION IN

unt1, William– Corpus Ch

entially replas that can be d productioncarbon dodecing the fluorane-adminis100 dilutionlipid productatments betwnd maximumcontrols. Str

or further str

5

N MICROA

m H. Wilson2

risti, 2Bigelo

ace significanconverted in

n by stressingcane and deprescent dye, Bstered culturen BODIPY, ation increase

ween days 6 m factor 5.25ressing cells ress-induced

ALGAE BY

2 and Peter Cow Laborato

ant fossil fuento renewabg Dunaliellapleting nitratBODIPY 50es during staand analyzede by the averand 13, after

5 for D. tertiovia these mlipid enhanc

DODECAN

Countway2 ory for Ocean

l consumptioble fuel for da tertiolecta,te amounts in05/515, and fationary phad for green frage factor 5r dodecane wolecta nitratethods resulcement inve

NE ADDITI

n Sciences

on levels. Thdiesel engine

Isochrysis gn the growthflow cytome

ase and nitratfluorescence 5.22 and maxwas adminise-limited trelt in increaseestigations.

ION AND

hese s: biodiesel.galbana, andh medium. etry. Strains te-limited by flow ximum tered, and

eatments ed lipid

d

Page 6: Bigelow Laborato ry for Ocean Sciences · PDF fileassimilation iatom Thala and one bac xleyi had lo ... ipid product tments betw d maximum controls. Str ... leting nitrat BODIPY 50

THE CynWill Therand rThe undenoveand fthrouwhenprodclaimphas

E RELATION

thia Liles, Eie Wilson, N

re is growingrenewable enlipids produ

erstand how el dye used tflow cytomeugh exponenn placed into

duction in somm. Isolation ase with no sig

NSHIP BET

EnvironmenNational Cen

g concern fonergy source

uced by algaegrowth relato detect alga

etry offers quntial to statioo new mediame algal straand treatmengnificant cha

TWEEN MIC

ntal Sciencenter for Mari

r climate che. Algae utile have been tes to lipid cal lipids by fualitative anonary phase. a to mimic laains, howevents of each gange in grow

CROALGAE

es, Universitine Algae an

ange as the Ulize energy fridentified as

content to hafluorescencealysis of lipiPavlova lut

ate lag phaseer, the additigrowth phasewth.

6

E GROWTH

ty of Californd Microbiot

United Statefrom the sun s a potential rvest algae a

e. This dye inid productivtheri has beee. It is reportion of glucose showed an

H PHASE AN

rnia, Riversta, Bigelow L

es struggles tand carbon source of bi

at optimal con combinatio

vity throughoen determineted that orgase or glycero

n increase in

ND LIPID C

side; Dr. PetLaboratory f

to find a cosdioxide, a m

iodiesel. It isonditions. BOon with epiflout the algal ed to have thanic carbon sol did not inclipid conten

CONTENT

te Countwayfor Ocean Sc

st-efficient, smajor greenhs therefore imODIPY 505/luorescence growth cycl

he highest lipsources boosclusively sup

nt only durin

y and Dr. ciences

sustainable, house gas. mportant to /515 is a microscopy le, from lag pid content st lipid pport this g late lag

Page 7: Bigelow Laborato ry for Ocean Sciences · PDF fileassimilation iatom Thala and one bac xleyi had lo ... ipid product tments betw d maximum controls. Str ... leting nitrat BODIPY 50

VARBIV EmiEarlh BivasurroDiarundethe eedulDamsampchardiatopositcouldamou

RIATIONVALVE AQ

ily Bell-Hoeham College

alve aquacultounding watrrhetic Shellferstand the reeffect of nutris and a bio-

mariscotta Ripled on a regacterized usioms. Detritaltively to nutrd prove harmunts.

NS IN PHYQUACUL

rth1, Cynthie1, Bigelow L

ture provideer. Harmful fish Poisoninelationship brient addition-fouling macver and incugular basis. Cing an imagil matter incrrient stimulamful to huma

YTOPLANLTURE IN

ia Heil2 and Laboratory f

s substrate falgal specie

ng (DSP), habetween cultns on the microalgal specubated in situChanges in ting-flow-cytreased on botation within an health an

NKTON CN COASTA

Nicole Poulfor Ocean Sc

for epibenthies, such as Prave been asstured bivalveicroalgal comcies, Ulva lau with and wthe phytoplantometer (Floth Ulva and the first 24 hd bivalve far

7

COMMUNAL MAIN

ton2

ciences2

ic microalgarorocentrumociated withes and their ammunities astuca. Ostrea

without nutrienkton comm

owCAM). Aloyster incubhours on botrms. Proroc

NITIES ASNE

ae and can alm lima, whichh bivalve aquassociated mssociated wia edulis and ents (Osmoc

munity on eacll communitibations over th substrates

centrum lima

SSOCIAT

lter nutrient dh produces ouaculture in

macro and mith the EuropU. latuca we

cote©) in Wech substrate ies were domtime. Pseud

s. This potena was not pre

TED WITH

dynamics inokadaic acidthe Gulf of M

microalgae, wpean Oyster,ere collectedest Boothbayover time w

minated by pdonitzschia rntially toxic sesent in sign

H

n the d causing Maine. To

we examined Ostrea d from the y Harbor and

were pennate responded species

nificant

d

Page 8: Bigelow Laborato ry for Ocean Sciences · PDF fileassimilation iatom Thala and one bac xleyi had lo ... ipid product tments betw d maximum controls. Str ... leting nitrat BODIPY 50

EF

GloborgaCDOuptakLakeEast fractRivedecrdiffeexamthe Ghigh

FFECTS OF

bal warming anic matter (COM, have beke, carbon ae on microbiBoothbay H

tionated chloer DHS additeases in cert

erent sourcesmined (bacteGulf of Mainher trophic le

F DISSOLVER

1

and changinCDOM) into

een shown toassimilation aial communiHarbor, and corophyll a. Ations within tain phytoplas documenteeria, phytoplane may potenevels.

ED HUMICRIVERS ON

Zakary J

Colby Colle

ng precipitato the Gulf ofo influence mand growth. ity compositcharacterizedAn initial 12the first 24 h

ankton groupd rapid (6 hrankton, or hentially signif

COMPOUNN MARINE M

Jaques1,2, Cy

ege, 2Bigelow

tion patternsf Maine. Dismicrobial gro

The effects ion was examd using flow

20 hr experimhours; with ips. A secondr) responses eterotrophs) ficantly impa

8

NDS FROMMICROBIA

ynthia Heil2,

w Laboratory

have increassolved humowth throughof DHS isolmined using

w cytometry, ment documeincreases in d experimentto additionsand DHS so

act microbia

M THE DAMAL COMMU

Nicole Pou

y for Ocean

ased the transmic substanceh trace metallated from th

g in situ incuimaging cyt

ented the comtotal chloropt examining s depending ource. Resulal community

MARISCOTTUNITIES.

lton2

Sciences

sport of rivees (DHS), thl chelation, ehe Kennebec

ubations of setometry (Flommunity resphyll and bathe effect ofupon comm

lts indicate cy compositio

TA AND KE

erine coloredhe major comeffects uponc River or Deawater fromowCAM), ansponse to Keacteria, as wef DHS addit

munity compochanging DHon, in turn e

ENNEBEC

d dissolved mponent of n nutrient amariscotta

m West and nd size-ennebec ell as tions from onent

HS levels in ffecting

Page 9: Bigelow Laborato ry for Ocean Sciences · PDF fileassimilation iatom Thala and one bac xleyi had lo ... ipid product tments betw d maximum controls. Str ... leting nitrat BODIPY 50

DETPHY And1Uni Cytoelectlongadap(Woanalyusingaminphysfor rproteto ad

TECTING EYSIOCHEM

drew Burchiiversity of C

ochrome b (ctron transpor

gevity, obesitptations. Conolley et al., 2ytical methog the physicno acid chansicochemicalobust interprein. This studaptation dur

EVOLUTIOMICAL SHI

ill1, David Mhicago, 2Big

cytb) is a conrt chains invty, and age-rnsequently, t2003), I ana

od has demonochemical p

nges and the l shifts in funretations of tdy suggests ring mamma

ONARY SELIFTS IN TH

McClellan2 gelow Labor

nserved protvolved in resprelated diseathere are fewlyzed cytb senstrated the

properties assproperties innctional protthese resultsthat several

alian evoluti

LECTION HE MAMMA

ratory for Oc

ein found inpiration and

ases like Alzhw mammal-wequences frosensitivity nsociated withnvolved in thtein regions s in terms of regions of thon.

9

IN HIGHLALIAN CY

cean Science

n nearly all euphotosynthe

heimer's. Mawide studies com 28 specienecessary to h structure ahe adaptive palso were de

f the electronhe protein th

LY CONSERYTOCHROM

es

ukaryotes anesis. Some hany statisticacharacterizines from nearcharacterizeand functionprocess wereescribed. Th

n transport anhought to be

RVED PROME B

nd prokaryothuman cytb mal models fang cytb evolrly all major e the evolution. The locatioe identified. he well-studind proton pu nonfunction

OTEINS:

tes, and is comutations afail to detect slution. Usingmammal lin

on of conseron of importDirectional

ied nature ofumping funcnal have bee

ommon in ffect subtle g TreeSAAPneages. This rved proteinstant cytb

f cytb allowsctions of the en important

P

s

s

Page 10: Bigelow Laborato ry for Ocean Sciences · PDF fileassimilation iatom Thala and one bac xleyi had lo ... ipid product tments betw d maximum controls. Str ... leting nitrat BODIPY 50

INDMela1Colb Atm97%unit Intergthencoccwere(280AddperiForaninfollo

IRECT EFFanie Ross1, Dby College, 2B

mospheric pC% by the end

decrease in governmentaneriticcalancolithophoreeallraised0ppm,380pitionally,deodsthrough280ppm,thncreasegrazowedby280

FECTS OF ODavid FieldsBigelow Labo

CO2 has risenof the centurthe oceans, pl Panel on Clnoidcopepoe,PleurochryatapCO2leppm,and75e‐platedPlehoutthe48hegrazingrazingrates.T0ppmand3

OCEAN ACIs2, Barney Boratory for Oc

n by 36% overy. The oceapredicted to imate ChangeodAcartiatorysis,weremvelof380p0ppm,reprurochrysis‐hourre‐plaatesdecreasThehighest380ppmresp

IDIFICATIOalch2, Steve cean Sciences

er the preindanic absorptidecline anot

e. Thisstudyonsa.Ingestmeasuredinpmwhilethresentingprfromallthratingprocessedduringtingestionrpectively.

10

ON ON ACAShema2, As

s, 3Lake Supe

dustrial levelion of that inther 0.3 to 0yinvestigattionratesofn24‐hourbohealgaewere‐industriareepCO2levsstocompathere‐platinatesforreg

ARTIA TONshley Poehlserior State Un

l and is expenflux of atmo0.46 pH unitstedindirectfA.tonsafeottleexperiregrownatal,current,avelswerefeareingestionngprocess,gularlyplate

NSA 3

niversity

ected to increospheric COs by 2100 acteffectsofoedingonthimentsat16tthreedistiandpredictedinthreesnrateswithwhile380aedcellswer

ease by an adO2 has causedccording to thoceanacidifiecoastal6ºC.ThecopnctpCO2levedCO2inpueparate4‐hhcells’litheand750ppmrefrom750p

dditional d a 0.1 pH he icationon

pepodsvelsts).hourgrazingstructure.mbothsawppm,

g

Page 11: Bigelow Laborato ry for Ocean Sciences · PDF fileassimilation iatom Thala and one bac xleyi had lo ... ipid product tments betw d maximum controls. Str ... leting nitrat BODIPY 50

EXPZETAlys

1TheEast Zetapiron formwhatsheasequsigna

PERIMENTTAPROTEOson Lowell1,

e University Boothbay H

proteobacteis oxidized,

m is largely ut is a largely

ath-forming Zuences to genatures. 375 1

TAL TECHNOBACTERIA

2, Dr. Erin F

of Maine, OHarbor, Main

eria (“Zetas”extracellula

unknown, hoy inorganic mZetaproteobanerate fluore16S rRNA Z

NIQUES FOA: A METHField2, Dr. D

Orono Mainene 04544

”) are marinear structures owever studiematrix of Fe-acteria. Inve

escent in situZetaproteoba

OR CHARAHODOLOGIDavid Emerso

e, 04469; 2Bi

e microorgansuch as helices have indic-oxides. Thisestigations c

u hybridizatioacteria gene

11

ACTERIZINICAL APPRon2 and Dr. D

igelow Labo

nisms that ocal stalks or cated that pos research focentered on ion (FISH) prsequences w

NG SHEATROACH David McCl

oratory for O

xidize iron itubular shea

olysaccharidocused on preidentifying crobes and to

were compilesourccompUnfoalignworkreproCompdetersheatorganprecibactepolysSpectranspsheatfreshtargepolys

TH-FORMI

lellan2

Ocean Scienc

in microaeroaths form. H

des are an orgeliminary at

characteristico identify noved from bothces representpilation of Zeortunately, dinment were ok must be donoducibility anparative gen

rmine possibth-forming "nisms represipitates and neria were usesaccharide exifically, a larporter genesth-forming B

hwater iron oeted as potensaccharide ex

ING

ces, 60 Bigel

ophilic envirHow these strganic compottempts to idc “Zeta” shevel genes ash public and ting the larg

Zetaproteobaiscrepanciesobserved andne to increasnd robustnes

nomics was uble genetic si"Zetas.” Eighsenting stalknon-structured to determxport genes.rge number s were identiBetaproteobaoxidizer. Fivntial genes inxport.

low Drive,

onments. Asructures onent of entify

eath-forming genetic in-house est cteria. in sequence

d additional se ss. used to ignatures forht novel s, sheaths, e forming

mine . of ABC ified in acteria, a e genes were

nvolved in

s

g

e

r

e

Page 12: Bigelow Laborato ry for Ocean Sciences · PDF fileassimilation iatom Thala and one bac xleyi had lo ... ipid product tments betw d maximum controls. Str ... leting nitrat BODIPY 50

Abstan adunitsthis sPleuconc

EFFECTS

tract: Atmosdditional 100s and IPCC pstudy we exa

urochrysis spcentrations o

S OF OCEANMORPH

Ash1 Lake Su

spheric pCO2

0% by the enpredictions samined the e

p. Growth ratof 280, 380, a

N ACIDIFICHOLOGY O

hley Poehls1

uperior State

2 levels havend of the censuggest an adeffects of incte, culture biand 750 ppm

CATION ONOF THE COC

, David FielUniversity,

e increased bntury. Risingdditional deccreased pCOiomass, and

m. SEM, bire

12

N GROWTHCCOLITHO

lds2, Barney 2 Bigelow L

by 36% over g pCO2 has acrease of 0.3

O2 levels on tcell biomas

efringence, a

H RATE, BIPHORE PLE

Balch2, StevLaboratory fo

r the last centalready caus3 to 0.45 pH the calcifyins were meas

and light micvisstruincandandcelimastruresphyaltecocconproproeco

OMASS, ANEUROCHRY

ven Shema2

or Ocean Sc

tury and aresed ocean pH

units by theng coccolithosured at calibcroscope imasualize pCO2

ucture. Datacreasing pCOd culture biod X respectivll biomass inages revealeucture with isults indicateysiologicallyered by oceaccolithophorntributors tooductivity, coperties, thesological imp

ND COCCOYSIS

iences

expected toH to decreasee end of the cophore algaebrated pCO2

ages were us2 effects on ca shows that O2 levels, groomass decreavely, while i

ncreased by Xed a compromincreasing pe that Pleuroy and morphan acidificatires are major the ocean’sarbon cycle,se effects ha

plications.

OLITH

o increase bye by 0.1 century. In e, sed to cell and lith with owth rate ased by X individual X. SEM mised lith

pCO2. These ochrysis are hologically ion. Becauser

s primary , and optical

ave many

y

e

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EXAMAR Pritc Nues Twin Moszoopinfluand bvacuphytfluor10 tidigeinvercoccquan

AMINING TRINA USIN

chard, K. R.,

ster, J., Bige

ning, B. S., B

t of the iron plankton grazuences of prebioavailabili

uoles of the htoplankton prescent dye times more ofst these cellsrsely propor

colithophoresntitatively de

THE GRAZNG A RATI

Northeaster

elow Laborat

Bigelow Lab

used by phyzers. Undersey-type on thity of regeneheterotrophicrey (diatom,to compare df a chlorophs. However, rtional to pres than with determine the

ZING PREFIOMETRIC

rn University

tory for Oce

boratory for

ytoplankton standing chehis processinerated iron. Ac dinoflagell, coccolithopdigestive vacyte than a coinitial fluore

ey preferencedinoflagellateffect of pre

FERENCESC FLUORES

y, Boston, M

ean Sciences

Ocean Scien

in high-nutremical proceng is thereforAs such, we late Oxyrrhisphore, chlorocuole acidityoccolithophoescence ratioe, with vacuotes. Additioney digestion

13

S AND DIGESCENT DY

MA, USA, ka

, East Booth

nces, East Bo

ient, low-chssing of ironre useful to ustudied the gs marina. Wophyte) overy with each pore (the leastos (with highole fluoresce

nal work willon iron spec

ESTIVE PRYE

aitlynpritcha

hbay, ME, U

oothbay, ME

hlorophyll aren within grazunderstandingrazing pref

We comparedr 24 hours. Inprey. Overalt consumed ph ratios correence ratios al constrain pciation.

ROCESSES

ard1@gmail.

USA, jnuester

E, USA, btw

eas comes frzer digestiveng grazer coferences andd grazing raten addition, wll, we found prey) and toesponding toapproximatelpH from thes

S OF OXYR

.com

[email protected]

wining@bige

rom reminere systems anntrols on the

d digestion ofes of three tywe used a ratthat Oxyrrh

ook 2-3 timeso low pH) wly 6 times hise ratios and

RRHIS

org

elow.org

ralization byd the e speciation f prey withinypes of tiometric is consumeds as long to

were igher with

d

n

d

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ThaFouNSFSiteOceResMai

anks to the Nundation for F Grant OCEe: Bigelow Lean Sciencessearch Experine and the W

14

National Scieits support uE 1156740 –

Laboratory fos – Undergrarience in the World Ocean

ence under – REU or aduate

Gulf of n.