The document summarizes a research project measuring nitrate levels at a wetland restoration site in Washington called Qwuloolt. A levee was breached to allow tidal flow from Ebey Slough. Samples were taken weekly from 6 locations and a slight upward trend in nitrate levels was observed, possibly due to nutrients from former farmland soil being transported into the slough. Nitrate levels remained below levels harmful to fish but will need ongoing monitoring to ensure the wetland does not become polluted. The results provide baseline data to track changes from the restoration project and determine if it successfully creates suitable salmon habitat.

1. The Concentration of Nitrite and its Role in the DissolvedOxygen Content in
Ebey Slough
BIS 499 Independent Research Project
Dr. Robert Turner
Tajinder Singh
6/8/2016
Abstract
As a part of a researchteam withclassmatesChrisThemelis andKabrinaC.Orchardat
the Universityof WashingtonBothell,we collectivelygatheredwaterqualitydatafrom
withinandnearthe QwulooltestuaryrestorationsiteinMarysville,WA.Afteralevee
breachin August2015 that allowedtidal inundationfromEbeysloughtoaformer
farmlandinhopestofacilitate natural hydrologicprocesses thatoccurwhenfreshand
saline watermix and eventuallyformanestuarine wetlandthatwillserve asasite for
salmonrearing.We mainlymeasurednitrate levelsatthe site usingthe SUNA senorand
trackedany fluctuations innitrate levels because of the breach. Althoughitistooearly
to tell at thisstage of the restorationprojecttoconclude anythingsubstantial,there
was a slightupwardtrendof nitrate levelsinandaroundthe breach site.This may
indicate thatnutrientsfromthe soil inthe formerfarmlandare beingdistributedinto
Ebeyslough.Thatis not a matterfor concern, butit if nitrate levelscontinue torise,
Ebeysloughmightbecome polluted withexcessnitrateand/orphosphorouscoming
fromboth surface runoff andthe soil, whichmaycause eutrophication andlowerthe
dissolvedoxygencontentof the water toa pointitmay nolongersustainaquatic
wildlife.

2. 1
Table of Contents
Introduction: pg. 2
Site Characteristics: pg. 4
Methods: pg. 9
Results: pg. 10
Discussion: pg. 12
Conclusion: pg. 14
References: pg. 16
Figures
Fig. 1: pg. 5
Fig. 2: pg. 6
Fig. 3: pg. 7
Fig. 4: pg. 7
Fig. 5: pg. 8
Fig. 6: pg. 10
Fig. 7: pg. 11
Fig. 8: pg. 11
Fig 9: pg. 12

3. 2
Introduction:
The main goal of the QwulooltEstuary RestorationProjectistorestore the natural processesof rivers
and tidesbybreachinga levee dividingEbeyslough froma400-acre formermarshland andfarmingarea
knowsas the Qwulooltfloodplain. A Estuarine residence like the Qwulooltfloodplain site whenrestored
wouldprovide juvenilesalmon withextensive forage opportunities,refuge frompredation, andtime for
physiological adaptationtoincreasingsalinitiesduringthe transitionfromfreshtosaltwater all of which
isessential forsurvival andgrowthof keystone PNWsalmon(Healey1982).
Thisrestoration projectinvolved cooperationfromthe following stakeholdersand representatives:
 TulalipTribesof Washington.
 National OceanicandAtmosphericAdministration(NOAA).
 U.S. Fishand Wildlife Service (USFWS).
 WashingtonDepartmentof Ecology(WDOE).
 US Army Corpsof Engineers.
 Natural Resource ConservationService(NRCS).
 Cityof Marysville.
Salmonare an integral part of the economyandecosystemof the PNWsince theyare a keystone
speciesandover137 differentspeciesdepend onsalmondirectlyandindirectly (WDOE2002). The long-
termgoal of the Qwulooltrestorationistotransformthe formerfarmlandintoaself-sustaining,
vegetatedestuarine wetlandthatwillmaximizethe natural ecologicalpotential of the site andfacilitate
the natural hydrologicprocesses,whichwill supportanddepositsedimentandseedsrequiredfor
successionof native vegetationandplantcover,all of which will facilitate inprovidingoptimalrearing
habitatfor salmongoingupstreamthroughEbeyslough.The floodplain iscurrently home tovarious
invasive speciessuch asreedcanarygrass, thistle,and Himalayanblackberry.One of the goalsisto

4. 3
promote diebackof these typesof invasive vegetationat the site inorderto provide native
reintroductionandplantcover.
The official objectives of the restoration include accordingtothe Qwulooltwebsiteisasfollows:
 To maximize cover,forage,migratorypathways,andotherhabitatfunctionsforsalmon.
 Facilitate native vegetationre-establishmentthroughrestorationplanting.
 Promote diebackof invasivevegetationcoverthrough brackishwaterre-introduction.
 Restore the publicandTulalippeople’sconnectiontothe Qwulooltmarsh.
 Protectinfrastructure byconstructingasetbacklevee.
 Enhance waterqualitythroughconstructionof stormwatertreatmentcells.
 Recoverthe natural streamand floodplainformsthrough fillingof drainage ditchesandre-
contouringof mainstreamanddistributarynetworks.
 Increase salmonpopulationsandchannel-floodplainhabitatfunctionsthroughplacementof
large woodwithincreeks.
Accordingto the SnohomishBasinSalmonConservationPlan(SnohomishBasinSalmonRecoveryForum
2005), the qualityandquantityof rearinghabitatinthe nearshore estuary andmainstreamriversisthe
primaryfactor limitingChinookandbull trout salmon.The Qwulooltrestoration projectisexpectedto
benefitthesefederallythreatened species,aswell as otherfishandwildlife byincreasingthe extentand
connectivityof estuarine wetlandsinthe Snohomish area.

5. 4
Site Characteristics
The Qwulooltfloodplainis354 acresin size,the oldleveewasbuiltabout110 years ago to preventtide
and saltwaterfrom comingintothe thenfarmland. The US ArmyCorps of Engineersbreached the levee
and dugout about 260 feetgapto allowtide tocome infromEbeyslough inhopesto create a mix of
saltand freshwatertopromote optimal rearinghabitatforsalmonanda diverse wetlandwithvarious
native species.
Our mainsamplingsitesfornitrate were asfollows:
 QC1 (QuilcedaCreekFishCatchSite)
 CR6 (Directlyinfrontof the Marysville sewagetreatment facility locatednearthe Qwuloolt
floodplain)
 CR5 (Atthe mouthof the "Marysville RestorationSite:JonesCreek")
 CR4 (Belowthe breach,atthe mouthof the EbeyFyke;NOAA'slowerfishcatchsite)
 CR3 (Locateddirectlyinfrontof the levee breachsite)
 CR2 (About30m above the breach)
 CR1 (PointFurthestabove breach.Justpastthe HeronFyke NOAA fishcatchsite)

6. 5
Figure 1. DepictingQwulooltrestoration siteaerial view(2009)
Source: (Monitoring Plan for the QwulooltRestoration Project. Rice, Casimir 2011)

7. 6
Figure 2. Blue indicates minor excavation,red is major excavationsand the yellowis location of new setback
levee. (Image source: http://www.qwuloolt.org/AboutUs)

8. 7
Figure 3. Pictureof the breach sitethe day before from a webcam installed atthe site.
Figure 4. At the siteof the breach on August 28 2015, the US Army Corps of Engineers excavating.

9. 8
(Image source: http://www.heraldnet.com/article/20150828/NEWS01/150829126)
Figure 5. Photo taken on August 28 2015 after the levee breach.

10. 9
Methods
We collecteddatafromthe Qwulooltrestorationsite onaweeklybasiseveryTuesday duringlow tide
from10/21/2015 until 11/28/2015. We sampledmainlyatthe pointsmentionedearlierinthe site
characteristicssection usingourequipment.Ourmethods includedgoingtothe site every Tuesday
duringlowtide toensure we were able togetclose enoughtoEbeySloughand the breachsite safely
since we were onfootand didnot have accessto a boat.We recordedthe datacollectedfromthe YSI 85
and SUNA sensor.
The YSI gave usimportantwaterquality parameterslikethe temperature,salinity,dissolvedoxygen
content,andconductivity levelsinthe water.We wantedtosee the connectionbetweendissolved
oxygencontentandnitrate since highnitrate concentrationscontributetoeutrophication,whichis
whenexcessgrowthof vegetationinthe watersuchas algae can occur, which inturn lowersthe
dissolvedoxygen content, whichfishandotheraquaticwildlifedependon.The SUNA (Submersible
UltravioletNitrate Analyzer) wasusedtotake bothfreshand salt-waternitrate readings,the SUNA was
developedtomeasure nitrogen-basednutrientsconcentrationsinocean,estuarine andhighturbidity
freshwaterenvironments.Totake nitrate readingswe wouldsubmerse the SUNA sensorinthe waterfor
at 15-25 secondstocollectthe readingsviathe attachedtabletandthensave it andexportit toa
MicrosoftExcel spreadsheetforgraphical analysis.
We alsoconductedbrief site surveysof the vegetation, andwritingdownanychangestothe site if
applicable,forexample newalgae growth,tide level,wind,weather,etc.

11. 10
Results:
Figure 6. Nitrate levelsfromfall 2015 were consistentbutshowedaslightupwardtrend,the graphin
thisfigure combined nitrate levelswithinfromEbeyslough.The sitesare CR1,CR2, CR3, CR4, CR5, and
CR6.
0
0.1
0.2
0.3
0.4
0.5
0.6
0.7
0 5 10 15 20 25 30 35 40
mg/Lnitrate
Day of data collection
Nitrate (mg/L) Fresh Water Deployment: Nitrate
concentrations over 2 months Fall 2015

12. 11
Figure 7. Thisfigure depictsthe concentrations of nitrate in(mg/L) overtime similartofigure 6,this
representationshowsthe concentrationsgraduallygoingup withthembeinghighestonourlastday of
data collection11/28/2015 with0.6073 mg/L nitrate.
Figure 8. Thisfigure showssitessampledandthe average nitrate concentrations in(mg/L) afterthe
breachsampleswere takenfromOctober2015 to November 2015. The mainsample sitesare within
Ebeysloughwiththe exceptionof QC1,which islocatedatthe boatlaunchsite awayfrom the slough.
Source(Themelis,2016).
0.472472928 0.463765527
0.607372479
0
0.1
0.2
0.3
0.4
0.5
0.6
0.7
10/27/2015 11/24/2015 11/28/2015
NitrateConcentration(mg/L)
Date of Sampling
AverageNitrateLevels within Ebey Slough
0.60091818
0.60577857
0.60287143
0.61121667
0.61375556
0.60207143
0.6081
0.59
0.595
0.6
0.605
0.61
0.615
QC1 CR6 CR5 CR4 CR3 CR2 CR1
NitrateConcentration(mg/L)
SamplingSite
Nitrate Fall Averages 2015@ Main Sampling Sites:

13. 12
Figure 9. Thisgraph shows the nitrate averagestakenafterthe breachinthe summer2015. Asyou
compare these numberstothe onesinFigure 8, youwill see there isanincrease innitrate
concentrationsforeachof the sitesfromthe monthsof Octoberto November.Thisprovesthatnitrate
concentrationswill increase overtime becauseof allowingtidal inundationinthe Qwulooltsite from
Ebeyslough.Source(Themelis,2016).
Discussion
Because salmongain 95% of theirbodymassin the oceanthen returnto freshwatertospawnanddie,
the marine-derivednutrientsthey transportcanbe substantial fornutrient-poorfreshwaterstreamsand
lakes(Naimanetal.2002, Schindleretal.2003, Janetski etal.2009). In this case, the salmonthatwill be
travelingthroughthe Qwulooltrestorationsitewill be bringingwiththemessential nutrientsthatmay
aidin creatingan optimal spawninghabitat withinthe estuarine wetland.
The nitrate concentrationaveragesforthe Qwulooltestuaryrestorationsitewere all below 1.0mg/L
and rangedfrom0.45-0.61 mg/L. These nitrate concentrationsare nota toxicity concern.Nitrate
concentrations of 5-10mg/l are mildlytoxicto eggsandfry of cutthroat and rainbow troutspecies.
Chinooksalmonare affectedat20 mg/Lnitrate levels, andCohosalmonare sufficientlyresistant
(Kincheloe et.al.1979). As a general guideline,nitrateatlevels around10mg/L insurface waterlimit
salmonrearingandspawning.
0.596555
0.5385256
0.523789182
0.511777778
0.56256
0.500916667
0.511695
0.499182625
0.44
0.46
0.48
0.5
0.52
0.54
0.56
0.58
0.6
0.62
QC1 CR6 CR5 CR4 CR3.5 CR3 CR2 CR1
NitrateConcentration(mg/L)
Site Location
Nitrate Averages Summer 2015 (including postbreach point
CR3.5)

14. 13
Nitrate andphosphorusare extremelyimportantparametersinwaterqualitybecause increasednitrate
concentrationscontribute toexcess vegetationgrowthandalgal bloomsinwaters.Accordingtothe
EPA, nutrientpollutionisone of America’smostwidespreadandchallengingenvironmental problems.
Eutrophicationmayoccur at the qwulooltsite andEbeyslough if excessnitrogenandphosphorusfrom
sourcessuch as fertilizersandsurface runoff getsintothe surface waters andthatwill provide favorable
habitatfor plantgrowth such as algae andduckweed.Algal bloomsatthe surface of water bodies
preventsunlightfromreaching deeperdepths,asaresult,the plantsthatreside there die, anddissolved
oxygenisdepleted byabsence of respiration.Decomposersthenbreak downthe deadplants,which
furtherdepletesoxygenlevels. Eventually if pollutantslike nitrogenandphosphorouscontinue toenter
the waterbody hypoxiawilloccur.Hypoxiaoccurswhenthe dissolvedoxygencontentinthe water
depletestoapointthat itno longercan support aquaticlife atall.Since the levee breachwasseparating
Ebeysloughfroma formerfarmland,the soil onthe landitself couldbe asource of nitrate from
fertilizersusedinthe past,andfromanimal andplant residue thatmay have depositednitrogenintothe
soil.
We believe thatnitrate concentrationswill rise overtime,asthe Qwulooltsite isfloodedovertime from
the tide cominginfrom Ebeyslough.The tide will serveasa mediumto remove andtransportnutrients
fromthe restorationsite anddistributeinEbeyslough.Fromcomparingthe datainfigure 9 and figure 8
it isclearthat nitrate levelsinandaroundthe breachsite wentupovertime since the breachoccurred,
we needtocontinue tomonitorthese changesinnitrate levelstoensure thatthe restorationproject
will be successful.
Water temperature isanotherfactorwhendeterminingsurvival and rearinghabitatforsalmonin
estuaries. Adultsalmonhave experiencedpoorsurvival whenexposedtowatertemperatures greater
than 60°F (15.5°C),it isimportantto note that adultsheldinwatertemperaturesgreaterthan 15.5°C
and lessthan3.3°C have lessviable eggsthanthose heldat3.3°C-15.5°C whichisthe optimal

15. 14
temperature (Marine,K.R.,& Cech,J.J. 2004). Inour results, the temperature recordingdidnotexceed
10°C and averagedaround5.0°C, whichiswithinthe givenrange of survival oroptimal rearinghabitat,
giventhattemperature isnota parameterof major concernat thispointwe didnot graphically
representitaswe didthe nitrate levels.
Conclusion
Thisprojectis a continuationof the one thatbeganinspring2015 prior to the levee breachtoseta
baseline of datathat will be usedin the future forhydrologystudentsatUWB. We gathereddata to get
a betterunderstandingbetweenthe connectionof nitrate anddissolvedoxygeninEbeyslough afterthe
breachof a levee thatwasseparatingthe sloughfrom the formerfarmland forover100 years and thus,
allowingtidal inundation.The ultimate goal istoallow freshandsaltwaterto mix and eventuallycreate
an estuarine wetlandwithabundantnativespecies thatwill provide plantcoverandforage
opportunitiesforsalmontravelingupstreamand create afavorable environmentfor spawning.We do
not have strongconclusive datafromthe Qwulooltestuaryrestorationsite, giventhe shortsince the
breach, it isdifficulttoassessanychangestothe water quality thatmayhave a detrimentalimpact. In
figures6 and7, youwill see that the average nitrate concentrationsshowedaslightupwardtrend,and
the highestrecordedconcentrationonnitrate beingatCR3 (directlyinfrontonbreachlocatedinEbey
slough) andCR4 (belowthe breachsite),whichare inclose proximitytoone another. Itisimportantto
note infigure 9 and 8, the average nitrate levelsgraduallywentupovertimesince the breachinsummer
2015. Thispositive trendconfirmsourhypothesisthatnitrate storages fromthe soil inthe farmlandwill
migrate slowlyintoEbeyslough.The increasedconcentrationof nutrientsbecause of tidal inundation
will intheorypromote anoptimal estuarinehabitatforPNWsalmonand provide afavorable
environmenttogrow native vegetation.We mustbe patient before expectingsalmonpopulationto
make a dramatic returnsince thisproject islong-term.We cancontinue togatherdata and create

16. 15
graphical representationsof the nitrate levels,dissolvedoxygencontent,temperature,salinity,etc.,and
create a database of those parameters toreferto.If we continue togatherdata fromand monitorthe
Qwulooltrestorationsite inthe nextcomingmonthsandyearscontinuously,onlythenwe may come
across some more reliable datainordertopredictthe future and success of the Qwulooltrestoration
project.

17. 16
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