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1
Impact of Cladophora mats on E. coliconcentrations in beach water
Colleen M. McDermott, D.V.M., Ph.D.Department of Biology and Microbiology
University of Wisconsin - Oshkosh Oshkosh, WI 54901
Phone - (920) 424-1100Fax - (920) 424-1101
E-mail –[email protected]://bact.uwosh.edu/
Source: “Unknown”
2
Overview• Increased Beach Monitoring• Increased Cladophora• Previous research shows Cladophora
can harbor large amounts of E. coli• Impact of Cladophora on beach water
quality?• Impact of Cladophora on pathogens?• What does it all mean?
What is Cladophora?
From WIDNR 2004
•Filamentous branched structure
•Rarely found pure in environment
•Mainly grows on hard surfaces
•Common in highly eutrophic waters
•Often forms large ‘mats’
3
6/24
/05
6/28
/05
7/6/
05
7/12
/05
7/26
/05
8/3/
05
8/9/
05
10M Lft of Mat10M Rt of Mat
10M Away5M Lft of Mat
5M Rt of Mat5M Away
1M Lft of Mat1M Rt of Mat
1M AwayMat Proper
0
500
1000
1500
2000
2500
E. coli MPN/100mL
Date
Murphy Park
10M Lft of Mat 10M Rt of Mat 10M Away 5M Lft of Mat 5M Rt of Mat5M Away 1M Lft of Mat 1M Rt of Mat 1M Away Mat Proper
UW Sea Grant Project:Cladophora and Beaches
2007,2008
4
Lakeside Park Beach and
Whitefish Dunes State Park,
Door County, WI
5
Sea Grant Methods•Collaborative project – UW-Oshkosh, U.-MN, USGS, and Racine Health Department•Investigate spatial distribution of E. coli around Cladophoramats and the spatial distribution within mats.•Investigate the occurrence of pathogens associated with mats.•Other questions related to genetic make-up of E. coli recovered within mats.
Shoreline
45 cm deep water3 m
Sampling protocol for Objective 1, Question 1 (Is there a difference in E. coli concentrations inside and outside the mat?). Oval = Cladophora mat, Star = random sample site location (approximate).
1 mShoreline
4 meters 4 meters
Sampling scheme for Objective 1, Question 2 (Is there an E. coli gradient within and outside the Cladophora mat?). Oval = mat, Stars = sampling locations.
45 cm deep
Whitefish Dunes
6
2007
Water samples underlying & adjacent to mat- WFD 2007
7
Water samples underlying and adjacent to mat- Lakeside 2007
• Elevated concentrations of E. coli found in water within mats
• Elevated concentrations of E. coli found in water adjacent to mats
• E. coli concentrations in water underlying mats increases with time mat is stranded in nearshore water
8
Water samples underlying and adjacent to mat- WFD 2007
Water samples underlying and adjacent to mat- Lakeside 2007
9
E. coli attached to mat biomass-WFD 2007
10
2008• Wanted to return to same beaches• Paucity of Cladophora for most of
summer• WFD= okay (3 Events)• Lakeside = little Cladophora accumulation• Added Newport Bay State Park Beach
♦ Mat was stationary for most of summer♦ Did not have movement of mat in and out♦ Mat material was “aged”
Random Samples - Whitefish Dunes for Event 1
0
1
2
3
4
5
6
8/5/ 2008 8/ 6/ 2008 8/ 7/ 2008
Da t e S a mpl e d
Random MatRandom Left WaterRandom Right Water
11
Random Samples - Newport Beach for Event 2
0
0.5
1
1.5
2
2.5
3
3.5
8/5/2008 8/6/2008 8/7/2008
Date Sampled
E. c
oli
MPN
/100
mL
Random MatRandom Left WaterRandom Right Water
• E. coli concentrations remained elevated in water underlying and adjacent to mats
• E. coli concentrations increased over time within a mat, although not as dramatic as 2007♦ Age of mat may influence ability of E. coli to
replicate and/or survive
12
Whitefish Dunes Cladophora Mat - E. coli MPN/100mL for Event 3
0
0.5
1
1.5
2
2.5
3
3.5
A B C D E F G H I J 1 2 3 4 5 6 7 8 9 10 J I H G F E D C B A
St aked Sampl e Locat i on
Lef t Wat er Out side Mat Wit hin Mat Right Wat er Out side Mat
E. coli Genetic Fingerprinting
• To determine if E. coli in mat is replicating or if new inputs of E. coli are occurring.
• Used the HFERP technique (fluorophore-enhanced, repetitive extragenicpalindromic-PCR)
• Jack knife and Manova analysis of fingerprints
13
WFD 2007 E. coli isolatesAll Mat Events
7/16-7/187/31-8/28/8-8/10
Lakeside 2007 E. coli isolatesAll Mat Events
7/16-7/188/1-8/38/8-8/10
14
WFD 2007 E. coli isolates Mat Event #1: MDS
7/16/20077/17/20077/18/2007
Fingerprinting Summary
• E. coli isolates from the same mat are similar from one day to next= replication
• “New” E. coli input also occurs day to day • E. coli isolates (from same beach) are not
similar from one mat to another
15
Cladophora impact on pathogens
Pathogens found in Cladophora mats
•Measured Salmonella, Shigella and Campylobacter (standard culture techniques) in water from mats and attached to Cladophora biomass
•Two of three sampling events in 2007 & 2008 in Door County
16
Pathogen Results
• Found no Salmonella, Shigella, or Campylobacter in water or associated with Cladophora in 2007
• Found Salmonella and Campylobacter in water underlying mat at Whitefish Dunes during 1 event in 2008
What we know
• Have never isolated Salmonella nor Shigellafrom water in Door County in previous years
• Have been able to isolate Campylobacter from water at beaches in previous years, not associated with Cladophora accumulations
• Will use more sensitive molecular techniques to try to identify pathogens in Summer 2009
17
• Other have consistently found pathogens in association with Cladophora mats in southern Lake Michigan (not in Door County) using molecular techniques
• Occurrence of pathogens in mats is likely a matter of input, rather than suitable habit
Conclusions• Cladophora harbors E. coli in high concentrations, allowing for survival and growth
within mats. Our data (2007 & 2008) support results published by earlier researchers.♦ High E. coli concentrations attached to mat biomass♦ High E. coli concentrations in water underlying mat♦ E. coli concentrations within mats increases as mats linger on the beach
• E. coli within mats appears to be “clonal” (i.e. it replicates within mats). More work is needed on the genetic make-up of E. coli found in the Cladophora mats.
• E. coli within mats negatively impacts water quality adjacent to mats.• Additional field data is needed on weather impacts and how mat movement may
impact beach health.• Pathogens may not survive to the extent that E. coli does in Cladophora mats and we
see no evidence of possible pathogen growth.• E. coli may not be the best indicator of recent fecal contamination of beach water• Removal of Cladophora mats from beaches should improve water quality by reducing
the number of E. coli in adjacent water
18
Acknowledgements• UW-Oshkosh “Beach Group” – Drs. Todd Sandrin and Bob
Pillsbury, Ms. Amy Vanden Heuvel, Mr. Ben Murphy, many other student researchers
• Collaborators:♦ Dr. Mike Sadowsky♦ Dr. Richard Whitman♦ Dr. Julie Kinzelman
• Partners:• Door County Soil and Water Conservation Dept. • Door County Health Department (Rhonda Kohlberg)
• Funding from:• UW Sea Grant• US EPA BEACH Act• UW-Oshkosh
Thank you!
19
= Water Sample
= Water & Cladophora Sample
Top ViewTop View
LW RWCM
1 2 3 4 5 6 7 8 9 10A B C D E F G H I J A B C D E F G H I J
20
A B C D E F G H I J 1 2 3 4 5 6 7 8 9 10 A B C D E F G H I J0
20000
40000
60000
80000
100000
120000
140000
160000
180000E
. col
i M
PN/1
00m
L
Staked Water Samples
Whitefish Dunes E. coli in Water - Event 2
Left w ater outside of mat Within mat Right w ater outside of mat
A B C D E F G H I J 1 2 3 4 5 6 7 8 9 10 A B C D E F G H I J0
200
400
600
800
1000
1200
1400
1600
1800
2000
E. c
oli
MPN
/100
mL
Staked Water Samples
Whitefish Dunes E. coli in Water - Event 2
Left w ater outside of mat Within mat Right w ater outside of mat
21
Random Samples - Whitfish Dunes
5591.4
9763.1
22979.7
21.31155.2
1863.6
14.5 21.8 3280
5000
10000
15000
20000
25000
7/16/2007 7/17/2007 7/18/2007
Date Sampled
E.co
li M
PN/1
00m
L
Random Mat Random Water Left Random Water Right
Lakeside Park Beach Cladophora CFUs/g Mean Concentrations for Events 1,2,&3
0
10,000
20,000
30,000
40,000
50,000
60,000
1 2 3 4 5 6 7 8 9 10 11 12
Location Within Mat
CFU
s/g
Event 1 Average Event 2 Average Event 3 Average
22
Whitefish Dunes Cladophora CFUs/g Mean Concentrations for Events 1,2,&3
0
50,000
100,000
150,000
200,000
250,000
300,000
1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17
Location Within Mat
CFU
s/g Event 1 Average
Event 2 AverageEvent 3 Average
Factors Affecting Growth
• Phosphorous is an essential nutrient♦ Fertilizers, manure, & urban storm water
• Zebra mussels
• Light
• TemperatureHarris, 2005.
23
E.coli Outside the Host —Secondary Habitat
• A battle for survival
• Low rate of survival
• Low probability of colonizing a new host
MAMMALIAN HOST
EXTERNAL ENVIRONMENT
Sunlight Low nutrients
PredationVariable temperature, pH and moisture
Adapted from Winfield and Groisman. 2003.
Potential Cladophora impact on E.coli Survival
• Block UV light
• Increased nutrients
• Moisture retention
• More stable microbial community
• Favors growth and survival of microbes!
MAMMALIAN HOST
EXTERNAL ENVIRONMENT
Sunlight Low nutrients
PredationVariable temperature, pH and moisture
Adapted from Winfield and Groisman. 2003.
24
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Lab Microcosm StudyNot easy to assess pathogen survival in environment.
•Used lake water & Cladophora from local lake•Looks at specific influence on E. coli and pathogens in a controlled setting.
♦Microbe + Water♦Microbe + Water and Cladophora
•Measured: ♦microbes free in water♦Microbes attached to Cladophora
Grow Light ( 24” 20W)
Open Container w/H2O and/or Cladophora
25
E. coli Survival in the Lab Microcosm
1.0E+0
0
1.0E+0
1
1.0E+0
2
1.0E+0
3
1.0E+0
4
1.0E+0
5
1.0E+0
6
1.0E+0
7
1.0E+0
8
1.0E+0
9
0 1 2 3 4 5 6 7 8 9 10 11 12 13 14 19 26 33 41 48
Elapsed Time In Days
CFU
/mL
Water Water + Cladophora Attached
1.0E+0
0
1.0E+0
1
1.0E+0
2
1.0E+0
3
1.0E+0
4
1.0E+0
5
1.0E+0
6
1.0E+0
7
1.0E+0
8
1.0E+0
9
0 1 2 3 4 5 6 7 8 9 10 11Elapsed Time In Days
Salm
onel
la C
FU/m
L
Water Water & Cladophora Attached
Salmonella Survival in the Lab Microcosm
26
Time Required for 50% Decline of Microbes in Lab Microcosms
DNR Observations vs Beach Monitoring•Measure Cladophora using a WI DNR scale and data obtained from routine beach monitoring in Door County, WI•Preliminary ‘look’ at what the observable Cladophorameant, if anything, to the data being obtained as part of a comprehensive monitoring program.•Regression analysis of observational data versus E. coli data.
Picture from WI DNR 2004.
27
Cladophora Distribution Study
Ranked the presence of Cladophora on the beach on a scale of:
None (0)Low (1)Moderate (2)High (3)
ScaleLow
Cladophoraminimally present in
patches on the beach.
ModerateCladophoranoticeably present on the beach
and a nuisance.
HighCladophoracovers beach in wind rows.
Slide adopted from WI DNR 2004.
Relationship Between Observation and Beach Monitoring
*Not possible to analyze due to lack of data.
0.5230.413Whitefish Dunes
0.7910.073Whitefish Bay
0.7400.112Sunset
0.0793.387Sturgeon Bay Rec. Canal
0.0434.659Sandy Bay
n/an/aSand Dune*
0.7670.092Rock Island
0.4410.611Portage Park
0.5450.384Percy Johnson
0.2671.257Otumba
0.0146.486Nicolet
0.4380.609Newport
0.1512.121Murphy
0.4170.678Lakeside
0.0434.957Jackson Harbor Ridges
0.2731.311Haines
0.5440.390Gislason
0.4990.464Fish Creek
0.6240.246Europe 3
0.3460.92Europe 2
0.8580.033Europe 1
0.4100.691Ephraim
0.7340.117Ellison
0.3850.766Egg Harbor
0.6790.173Bailey's Harbor
0.3151.047Anclam
P valueF valueBeach
0.5750.319Whitefish Dunes
0.1771.869Whitefish Bay
0.9010.016Sunset
0.8480.037Sturgeon Bay Rec. Canal
0.1791.855Sister Bay
0.9260.009Schoolhouse
0.2521.382Sandy Bay
0.8400.042Sand Dune
0.6370.234Rock Island
0.6960.156Portage Park
0.1093.041Percy Johnson
0.9620.002Otumba
0.0763.281Nicolet
0.8500.036Newport
0.1442.192Murphy
0.8840.022Lakeside
0.9000.016Jackson Harbor Ridges
0.2471.367Haines
0.5410.397Gislason
0.7910.071Fish Creek
0.4870.491Europe 3
0.9690.002Europe 2
0.9690.002Europe 1
0.2151.572Ephraim
0.7370.114Ellison
0.0136.621Egg Harbor
0.4130.681Bailey’s Harbor
0.6950.156Anclam
P valueF valueBeach
2005
2006
28
1.0E+0
0
1.0E+0
1
1.0E+0
2
1.0E+0
3
1.0E+0
4
1.0E+0
5
1.0E+0
6
1.0E+0
7
1.0E+0
8
0 1 2 3 4 5 6 7 8 9 10 11
Elapsed Time In Days
Shi
gella
CFU
/mL
Water Water + Cladophora Attached
Shigella Survival in the Lab Microcosm
Whitefish Dunes-Event 1
5591.4
9763.1
22979.7
21.31155.2
1863.6
14.5 21.8 3280.00E+00
5.00E+03
1.00E+04
1.50E+04
2.00E+04
2.50E+04
7/16/2007 7/17/2007 7/18/2007
E. c
oli
MP
N/1
00m
L
Random Mat Random Water Left Random Water Right
29
Whitefish Dunes – Event 2
1.00E+00
1.00E+01
1.00E+02
1.00E+03
1.00E+04
1.00E+05
1.00E+06
A B C D E F G H I J 1 2 3 4 5 6 7 8 9 10 A B C D E F G H I J
E. c
oli
MPN
/100
mL
Left water outside of matWithin mat Right water outside of mat
1.00E+00
1.00E+01
1.00E+02
1.00E+03
1.00E+04
1.00E+05
1.00E+06
A B C D E F G H I J 1 2 3 4 5 6 7 8 9 10 A B C D E F G H I J
E. c
oli
MPN
/100
mL
Left water outside of mat Right water outside of matWithin mat
Lakeside Park Beach – Event 3
30
Uses for stranded Cladophora?
What can be done with it, once it strands on my beach?-Composting-Use as pelleted heating product-Ethanol production-Methane production-Biofuel-And the list goes on…..
What we know now…
Presence of contaminating heavy metals or organics in Cladophora= very low-BTU potential= similar to White Pine-Composting project in progress in Ephraim-Use in methane digestor in progress in Manitowoc County
31
WFD 2007 E. coli isolates Mat Event #1
7/16/20077/17/20077/18/2007