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UnderstandingSourcesofContaminationinStormwaterPondstoPromoteWaterReuse

inAlbertaMeganBeaudry1,GrahamBanting1,Candis Scott1,NicholasAshbolt 1,

CaterinaValeo 2,Jianxun He3,NormaRuecker 4,BertvanDuin 4,Byeonghwa Jeon1,NormanNeumann1

1SchoolofPublicHealth,UniversityofAlberta,Edmonton,Canada.2 UniversityofVictoria,Victoria,Canada.3 UniversityofCalgary,Calgary,Canada

4 CityofCalgary,CalgaryCanada

1

Overview

• Waterreuse• ResearchObjective• Stormwaterpondmakeup• Molecularmethods• Culturemethods• McCallLakeResults

2

Overview1.WhyStormwater?

2.Sampling- WheredoIgetstormwater?

3.Methods- Howdoweanalyzestormwater?

4.Results-What’sinthewater?

5.Conclusion-Whatdoesthismeanforwaterre-use?

Alberta….Isthatastate?

3

Stormwaterisnotrainwater!

•Stormwaterisprecipitationcollectedafterreachingtheground.•Stormwaterpondscollectexcessrunoffincitiestocontrolfloodingandmanageurbanpollution.•HarvestingstormwatercouldprovideAlbertawithastrategytoaddressthegrowingdemandsonwaterresourcesduetoclimatechangeandprojectedpopulationgrowth.•Fromnuisancetoresource!

4

StormwaterUseOptions

• Wateringcommunitygardens/athleticfields• Toilet/urinalflushing• Clotheswashing• Firefighting• Waterfeatures

5

Whycan’twejustusethewater?!

• Potentialcontaminants:ØMicrobialpathogensfrom humanandanimalfecesØ Helminths,protozoan

• Stormeventshavebeenlinkedtoanincreasedincidenceofwaterborneentericdiseases,largelyduetomobilizationandtransportofpathogens.1,2

• Publichealthrisksassociatedwithstormwaterreusearenotwellunderstood.

1.Curriero etal.(2001).AmericanJournalofPublicHealth. 91:1194-9.

2.Thomas etal.(2006).InternationalJournalofEnvironmentalHealthResearch.6

So,what’sthepoint?

• Identifysourcesofbacterialcontaminationandbacterialpathogensinstormwaterpondsin

Calgary,Canada

7

821Weeks

CollectionofStormwaterSamples

PondA PondB PondC

MolecularMethods:WaterSamplesCollected

Filter20mLwaterthrougha0.4µmfiltertocapturebacteria

Culture100mLofwaterinQuantitrays®withColilert®forE.coli

detection(24h)

ExtractDNAfromfilter(EPAMethod

1611)

DNAextractedfromE.colipositivewells

QuantitativePolymeraseChainReaction(qPCR)

9

Whatcontributestothepond?qPCRTargets:MicrobialSourceTracking

- HF183 - Rum2Bac-Dog3- HumM2

- CGO1 - MuBac -LeeSg10

Whatinthepondcouldbearisktohumanhealth?qPCRTargets:IndicatorsandPathogens

Salmonella (InvA ) Campylobacterspp.(23S)STEC(stx1&2)- Dog72

Arcobacter butzleri (hsp60) Enterococcus spp.(Entero 1)- Dog72 11

Whyculture?Campylobacter andArcobacterMPNqPCREnumeration

30∘c &42∘c,

1248hours

Enterococcus andE.coliEnumeration

• Colilert– measurestotalcoliformsandE.coli

• Enterolert – detectsenterococci

13

OutfallML1

OutfallML2

Inlet3/4

Whatisastormwaterpond….

InletPR60

14

15

OutfallML2

OutfallML1

McCallLake

Area(ha) PercentResidential 537.38 37%

Commercial 99.84 7%

Industrial 294.70 20%

ParksandInstitutions

141.99 10%

MajorInfrastructure/Transportation

343.37 23%

FutureUrbanDevelopment

47.07 3%

Total 1464.35 100%

OutfallML1Area(ha) Percent

Residential 11.5 4%

Commercial 33.31 12%

Industrial 189.51 69%

ParksandInstitutions

30.54 11%

MajorInfrastructure/Transportation

11.48 4%

FutureUrbanDevelopment

0 0%

Total 275.99 100%

OutfallML2

16

PathogenFrequency

0

10

20

30

40

50

60

70

Campylobacterspp.

Salmonellaspp. Arcobacterbutzleri STEC

Percen

tsam

plespositive

Pathogen

N=364

17

DoCultureandMolecularmethods compare?qPCRforA.butzleri

Positive NegativeCultureforA.butzleri

Positive 6 18Negative 0 8

qPCRforCampylobacter spp.

Positive NegativeCultureforCampylobacterspp.

Positive 0 0Negative 0 32

18• 65%ofsamplescontainedA.butzleri throughmolecularmethods,howeverthismaybeashighas75%basedonculture

Whocontaminatedthestormwaterpond?HostFecalSource Marker Frequencyof

Occurrence (n=533)Human HF183 149(0.28)

HumM2 56(0.11)

Seagull LeeSg 48(.09)

Dog Dog3 13(.02)

CanadaGoose CG01 12(.02)

Ruminant Rum2Bac 11(.02)

Muskrat Mubac 6(.01)19

0

1

2

3

4

5

6

0

1

2

3

4

5

69-May

16-M

ay

23-M

ay

30-M

ay

6-Jun

13-Ju

n

20-Ju

n

27-Ju

n

4-Jul

11-Ju

l

18-Ju

l

25-Ju

l

1-Au

g

8-Au

g

15-Aug

22-Aug

29-Aug

5-Sep

12-Sep

Log 1

0CFUpe

r100

ml

Log 1

0MPN

per100

ml

DateSampled

E.coli

OutfallML2– E.coli

20

0

1

2

3

4

5

6

0

1

2

3

4

5

69-May

16-M

ay

23-M

ay

30-M

ay

6-Jun

13-Ju

n

20-Ju

n

27-Ju

n

4-Jul

11-Ju

l

18-Ju

l

25-Ju

l

1-Au

g

8-Au

g

15-Aug

22-Aug

29-Aug

5-Sep

12-Sep

Log 1

0MPN

per100m

l

Log 1

0Co

piesper100

ml

DateSampled

E.coli HF183OutfallML2– HumanMarkers

21

0

1

2

3

4

5

6

0

1

2

3

4

5

69-May

16-M

ay

23-M

ay

30-M

ay

6-Jun

13-Ju

n

20-Ju

n

27-Ju

n

4-Jul

11-Ju

l

18-Ju

l

25-Ju

l

1-Au

g

8-Au

g

15-Aug

22-Aug

29-Aug

5-Sep

12-Sep

Log 1

0M

PNper100m

l

Log 1

0Co

piesper100

ml

DateSampled

HumM2 E.coli HF183

OutfallML2– HumanMarkers

22

OutfallML2– A.butzleri

0

1

2

3

4

5

69-May

16-M

ay

23-M

ay

30-M

ay

6-Jun

13-Ju

n

20-Ju

n

27-Ju

n

4-Jul

11-Ju

l

18-Ju

l

25-Ju

l

1-Au

g

8-Au

g

15-Aug

22-Aug

29-Aug

5-Sep

12-Sep

Log 1

0Co

piesper100

ml

DateSampled

HF183 HumM2 HSP60

23

WhatistherelationshipbetweenA.butzleri andHumancontaminationmarkers?

qPCRforA.butzleri

Positive NegativeqPCR forHF183

Positive 14 24Negative 0 3

qPCRforA.butzleri

Positive NegativeqPCR forHumM2

Positive 9 5Negative 5 18

24

0

1

2

3

4

5

6

0

1

2

3

4

5

69-May

16-M

ay

23-M

ay

30-M

ay

6-Jun

13-Ju

n

20-Ju

n

27-Ju

n

4-Jul

11-Ju

l

18-Ju

l

25-Ju

l

1-Au

g

8-Au

g

15-Aug

22-Aug

29-Aug

5-Sep

12-Sep

Log 1

0MPN

per100

ml

Log 1

0Cop

iesp

er100

ml

DateSampled

LeeSg HSP60 E.coli

OutfallML2- Seagull

25

0

1

2

3

4

5

69-M

ay

16-M

ay

23-M

ay

30-M

ay

6-Jun

13-Jun

20-Jun

27-Jun

4-Jul

11-Jul

18-Jul

25-Jul

1-Aug

8-Aug

15-Aug

22-Aug

29-Aug

5-Sep

12-Sep

Copiesper100m

l

DateSampled

HF183 HumM2 LeeSg HSP60

OutfallML1– AbsenceofHuman

Markers

26

27

Whatarethefindings?• HumansewageappearstobeasourceoffecalpollutioninstormwatersystemsinMcCallLakeØdilutedlevels(~10-3)atoutfalls.

• A.butzleri,wasthemostprevalentbacterialentericpathogenfoundinstormwater(~65%)• A.butzleri sourceslikelyincludehuman,animals,andgrowthintheenvironmentasanimals(i.e.,seagullsandbirds).

28

Whatdoesthismean?

•Quantitativemicrobial risk assessment (QMRA)•Characterizing infrastructure challengesassociated with humanfecal pollution•Assess occurrence of other pathogens instormwater

29

Acknowledgements

• ProvincialLaboratory• EnvironmentalHealthSciencesGroup

30

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