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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
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Overview
• Waterreuse• ResearchObjective• Stormwaterpondmakeup• Molecularmethods• Culturemethods• McCallLakeResults
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Overview1.WhyStormwater?
2.Sampling- WheredoIgetstormwater?
3.Methods- Howdoweanalyzestormwater?
4.Results-What’sinthewater?
5.Conclusion-Whatdoesthismeanforwaterre-use?
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Alberta….Isthatastate?
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Stormwaterisnotrainwater!
•Stormwaterisprecipitationcollectedafterreachingtheground.•Stormwaterpondscollectexcessrunoffincitiestocontrolfloodingandmanageurbanpollution.•HarvestingstormwatercouldprovideAlbertawithastrategytoaddressthegrowingdemandsonwaterresourcesduetoclimatechangeandprojectedpopulationgrowth.•Fromnuisancetoresource!
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StormwaterUseOptions
• Wateringcommunitygardens/athleticfields• Toilet/urinalflushing• Clotheswashing• Firefighting• Waterfeatures
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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
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So,what’sthepoint?
• Identifysourcesofbacterialcontaminationandbacterialpathogensinstormwaterpondsin
Calgary,Canada
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821Weeks
CollectionofStormwaterSamples
PondA PondB PondC
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MolecularMethods:WaterSamplesCollected
Filter20mLwaterthrougha0.4µmfiltertocapturebacteria
Culture100mLofwaterinQuantitrays®withColilert®forE.coli
detection(24h)
ExtractDNAfromfilter(EPAMethod
1611)
DNAextractedfromE.colipositivewells
QuantitativePolymeraseChainReaction(qPCR)
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Whatcontributestothepond?qPCRTargets:MicrobialSourceTracking
- HF183 - Rum2Bac-Dog3- HumM2
- CGO1 - MuBac -LeeSg10
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Whatinthepondcouldbearisktohumanhealth?qPCRTargets:IndicatorsandPathogens
Salmonella (InvA ) Campylobacterspp.(23S)STEC(stx1&2)- Dog72
Arcobacter butzleri (hsp60) Enterococcus spp.(Entero 1)- Dog72 11
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Whyculture?Campylobacter andArcobacterMPNqPCREnumeration
30∘c &42∘c,
1248hours
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Enterococcus andE.coliEnumeration
• Colilert– measurestotalcoliformsandE.coli
• Enterolert – detectsenterococci
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OutfallML1
OutfallML2
Inlet3/4
Whatisastormwaterpond….
InletPR60
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OutfallML2
OutfallML1
McCallLake
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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
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PathogenFrequency
0
10
20
30
40
50
60
70
Campylobacterspp.
Salmonellaspp. Arcobacterbutzleri STEC
Percen
tsam
plespositive
Pathogen
N=364
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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
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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
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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
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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
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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
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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
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WhatistherelationshipbetweenA.butzleri andHumancontaminationmarkers?
qPCRforA.butzleri
Positive NegativeqPCR forHF183
Positive 14 24Negative 0 3
qPCRforA.butzleri
Positive NegativeqPCR forHumM2
Positive 9 5Negative 5 18
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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
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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
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Whatarethefindings?• HumansewageappearstobeasourceoffecalpollutioninstormwatersystemsinMcCallLakeØdilutedlevels(~10-3)atoutfalls.
• A.butzleri,wasthemostprevalentbacterialentericpathogenfoundinstormwater(~65%)• A.butzleri sourceslikelyincludehuman,animals,andgrowthintheenvironmentasanimals(i.e.,seagullsandbirds).
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Whatdoesthismean?
•Quantitativemicrobial risk assessment (QMRA)•Characterizing infrastructure challengesassociated with humanfecal pollution•Assess occurrence of other pathogens instormwater
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Acknowledgements
• ProvincialLaboratory• EnvironmentalHealthSciencesGroup
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