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 –mcdermot@uwosh.eduhttp://bact.uwosh.edu/

Source: “Unknown”

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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’

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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

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Lakeside Park Beach and

Whitefish Dunes State Park,

Door County, WI

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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

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2007

Water samples underlying & adjacent to mat- WFD 2007

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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

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Water samples underlying and adjacent to mat- WFD 2007

Water samples underlying and adjacent to mat- Lakeside 2007

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E. coli attached to mat biomass-WFD 2007

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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

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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

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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

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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

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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

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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

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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

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• 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

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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!

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= 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

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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

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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

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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.

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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.

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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

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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

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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.

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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

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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

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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

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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

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WFD 2007 E. coli isolates Mat Event #1

7/16/20077/17/20077/18/2007