Prepared in cooperation with the Ohio Water Development Authority and 

Muskingum Watershed Conservancy District

Microbial Source Tracking Markers at Three Inland 

Recreational Lakes in Ohio, 2011

Open-File Report 2012–1222

U.S. Department of the Interior

U.S. Geological Survey
Cover image: View of area near Crystal Beach, Buckeye Lake, Ohio 

(photograph by Donna Francy, U.S. Geological Survey).
Microbial Source Tracking Markers 

at Three Inland Recreational Lakes 

in Ohio, 2011

By Donna S. Francy and Erin A. Stelzer

Prepared in cooperation with the Ohio Water Development Authority and 

Muskingum Watershed Conservancy District

Open-File Report 2012–1222

U.S. Department of the Interior

U.S. Geological Survey
U.S. Department of the Interior

KEN SALAZAR, Secretary

U.S. Geological Survey

Marcia K. McNutt, Director

U.S. Geological Survey, Reston, Virginia: 2012

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reproduce any copyrighted materials contained within this report.

Suggested citation:

Francy, D.S., and Stelzer, E.A., 2012, Microbial source tracking markers at three inland recreational lakes in Ohio, 

2011: U.S. Geological Survey Open-File Report 2012–1222, 8 p.
iii

Contents

Abstract...........................................................................................................................................................1

Introduction ....................................................................................................................................................1

Methods...........................................................................................................................................................1

Sampling Sites and Procedures .........................................................................................................1

Microbial Source Tracking (MST) Marker Analyses ......................................................................3

Escherichia coli and MST Markers in Lake-Water Samples .................................................................4

References Cited............................................................................................................................................8

Figures

 1. Map showing locations of the three study lakes in Ohio .......................................................2

 2. Graph showing percentages of samples with not detected, estimated 

quantities, and measured quantities of microbial source tracking markers: 

A, Buckeye Lake Crystal and Fairfield. B, Atwood Lake. C, Tappan Lake 

Main and ditch, 2011 ....................................................................................................................7

Tables

 1. Beach study sites and agencies responsible for sampling ...................................................3

 2. Standard curve characteristics for microbial source tracking (MST) markers .................4

 3. Quality-control field blank data for Escherichia coli and microbial source 

tracking (MST) markers, 2011 .....................................................................................................4

 4. Quality-control concurrent replicate data for Escherichia coli and microbial 

source tracking (MST) markers, 2011 .......................................................................................5

 5. Concentrations of microbial source tracking (MST) markers in inland lake 

samples, 2011 ................................................................................................................................6
iv

Conversion Factors

Multiply By To obtain

Length

foot (ft) 0.3048 meter (m)

Volume

liter (L) 0.2642 gallon (gal)

milliliter (mL) 0. 03381 fluid ounce (oz) 

microliter (μL) 1,000 milliliter (mL)

Mass

gram (g) 0.03527 ounce, avoirdupois (oz)

Temperature in degrees Celsius (°C) may be converted to degrees Fahrenheit (°F) as follows:

°F=(1.8×°C)+32
Microbial Source Tracking Markers at Three Inland 

Recreational Lakes in Ohio, 2011

By Donna S. Francy and Erin A. Stelzer

Abstract

During the 2011 recreational season, samples were 

collected for E. coli and microbial source tracking (MST) 

marker concentrations to begin to understand potential sources 

of fecal contamination at three inland recreational lakes in 

Ohio—Buckeye, Atwood, and Tappan Lakes. The results from 

32 regular samples, 4 field blanks, and 7 field replicates collected at 5 sites are presented in this report. At the three lakes, 

the ruminant-associated marker was found most often (57–73 

percent of samples) but at estimated quantities, followed by 

the dog-associated marker (30–43 percent of samples). The 

human-associated marker was found in 14 and 50 percent of 

samples from Atwood and Tappan Lakes, respectively, but was 

not found in any samples from the two Buckeye Lake sites. 

The gull-associated marker was detected in only two samples, 

both from Tappan Lake. 

Introduction 

Identifying sources of fecal contamination at recreational 

beaches with elevated Escherichia coli (E. coli) concentrations is important for implementing appropriate remedial and 

control strategies and understanding human health risk of the 

water’s use. Microbial source tracking (MST) is a term used 

for identifying sources of fecal contamination in the environment. MST techniques are based on the concept that intestinal 

systems of different warmblooded animals select for specific 

microbial populations in response to differences in diet and 

physiology. A widely used MST method is the use of hostassociated MST markers. For this method, a targeted genetic 

sequence of DNA is amplified by the polymerase chain reaction (PCR) into an amount that can be quantified. Host-associated MST markers have been identified from different groups 

of fecal bacteria, often from the genus Bacteroides, a bacterium abundant in the gut of warmblooded animals (Layton 

and others, 2006). Although the occurrence of host-associated 

MST markers has been investigated at coastal beaches (Bower 

and others, 2005; Wong and others, 2009; Edge and others, 

2010), to our knowledge there are no published studies that 

use these markers to identify sources of fecal contamination at 

inland recreational beaches. Inland water bodies are popular 

swimming and boating destinations throughout the United 

States and, because of differences in size and hydrology, the 

water quality at inland lakes may be affected differently by 

contaminant sources than at coastal beaches. 

This report describes the results of research by the 

U.S. Geological Survey (USGS), in cooperation with the Ohio 

Water Development Authority and the Muskingum Watershed Conservancy District, to begin to understand the sources 

behind elevated E. coli concentrations at three inland recreational lakes in Ohio. Thirty-two regular samples, four field 

blanks, and seven replicates were collected and analyzed for 

concentrations of E. coli, general Bacteroides, and host-associated markers for humans, ruminants, dogs, and gulls. The 

results are presented in this report. This work is part of a larger 

monitoring study that was done to determine whether predictive models for E. coli could be used to provide near-real-time 

assessments of water quality at inland recreational beaches. 

Methods

Sampling Sites and Procedures

Samples were collected from five sites at three inland 

recreational lakes in Ohio (fig. 1, table 1) affected by elevated 

E. coli concentrations. Buckeye Lake is a shallow manmade 

reservoir in central Ohio constructed in the early 19th century 

for the Miami and Erie Canal, which connected the Ohio River 

with Lake Erie. Two beach sites were included at Buckeye 

Lake—Crystal and Fairfield. Sources of fecal contamination to 

Buckeye Lake are largely unidentified and may include birds 

and other wildlife, runoff from parking lots, and agricultural 

animals. Atwood Main and Tappan Main are sites at popular 

swimming beaches on reservoirs located in mixed recreational 

and agricultural areas in eastern Ohio. Potential sources of 

fecal contamination at Atwood Main include birds and discharge from a nearby intermittent stream. Potential sources of 

fecal contamination at Tappan Main include birds and other 

wildlife, runoff from the parking lot, and a lagoon wastewater 

treatment system that discharges effluent into a nearby ditch. 

The ditch drains into Tappan Lake near the beach. A sampling 
ROSS

PIKE

WOOD

KNOX

STARK

DARKE LICKING

ADAMS SCIOTO

WAYNE

MIAMI

UNION

HURON

GALLIA

SENECA

MEIGS

LOGAN

LORAIN

BROWN

ALLEN

ATHENS

HARDIN

ASHTABULA

HENRY

CLARK

PERRY

NOBLE

BUTLER

TRUMBULL

PUTNAM

BELMONT

VINTON

FRANKLIN

MERCER

ERIE

MEDINA

MUSKINGUM

PREBLE

HIGHLAND

HANCOCK

SUMMIT PORTAGE

FULTON

SHELBY

LUCAS

MONROE

MARION

FAIRFIELD

HOLMES

MADISON

PICKAWAY

GREENE

RICHLAND

GEAUGA

CLINTON

GUERNSEY

MORGAN

WASHINGTON

FAYETTE

WARREN

HOCKING

LAKE

JACKSON

WILLIAMS

COSHOCTON

MORROW

CARROLL

PAULDING

DELAWARE

DEFIANCE

TUSCARAWAS

CUYAHOGA

AUGLAIZE

HAMILTON

LAWRENCE

WYANDOT

COLUMBIANA

VAN WERT

HARRISON

MAHONING

SANDUSKY

CHAMPAIGN

CRAWFORD

OTTAWA

ASHLAND

CLERMONT

JEFFERSON

MONTGOMERY

BUCKEYE

LAKE

TAPPAN

LAKE

ATWOOD

LAKE

0 20 40 MILES

0 20 40 KILOMETERS

81°0'0"W

81°0'0"W

82°0'0"W

82°0'0"W

83°0'0"W

83°0'0"W

84°0'0"W

85°0'0"W 84°0'0"W

42°0'0"N 42°0'0"N

41°0'0"N 41°0'0"N

40°0'0"N 40°0'0"N

39°0'0"N 39°0'0"N

2 Microbial Source Tracking Markers at Three Inland Recreational Lakes in Ohio, 2011

Figure 1. Locations of the three study lakes in Ohio.
Methods 3

Table 1. Beach study sites and agencies responsible for sampling.

[MWCD, Muskingum Watershed Conservancy District, New Philadelphia, Ohio; USGS, U.S. Geological Survey, 

Columbus, Ohio]

Site name USGS site identification 

number1 Short name Site description Sampled by

Atwood Lake Main Beach 403219081155500 Atwood Main Popular bathing beach MWCD

Buckeye Lake Fairfield 395520082281500 Buckeye Fairfield Small beach USGS 

Buckeye Lake Crystal 395557082283800 Buckeye Crystal Small beach USGS 

Tappan Lake Main Beach 401926081105100 Tappan Main Popular bathing beach MWCD

Tappan Lake ditch 401910081111200 Tappan ditch Ditch MWCD

1

Corresponds to the latitude and longitude of each site.

site, Tappan ditch, was included to compare the sources of 

fecal contamination in the ditch to those at the swimming 

beach. The potential for human sources from wastewater at 

Buckeye Lake sites and at Atwood Main are unlikely because 

the communities around these reservoirs are served by sanitary 

sewer systems.

Data were collected during July–September 2011 by the 

USGS Columbus office (Buckeye Lake sites) or by the Muskingum Watershed Conservancy District (MWCD, Atwood and 

Tappan sites). Thirty-two samples for E. coli and MST markers were collected by using standard grab-sampling techniques 

(Myers and others, 2007) at 2–3 ft water depths, in areas used 

for swimming. A 1-L or 3-L sterile polypropylene sample 

bottle was filled with water about 1 ft below the water surface, 

placed on ice, and transported by car or shipped overnight to 

the USGS Ohio Water Microbiology Laboratory (OWML) in 

Columbus, Ohio. In addition to regular sampling for E. coli

and MST markers, four field blanks and seven replicates were 

collected and analyzed. If a MST marker was detected in one 

replicate and not the other, the result from the positive replicate was used; otherwise, an average of two replicates was 

used for data analysis. 

Samples for E. coli were analyzed within 6 hours of collection at the OWML or MWCD office by use of the Colilert Quanti-Tray/2000 method (IDEXX Laboratories, Inc., 

Westbrook, Maine) and are reported in most probable number 

per 100 milliliters (MPN/100 mL). Sample processing and 

quality-control procedures are described in Myers and others 

(2007). Daily data on E. coli concentrations and MST markers were entered into the USGS National Water Information 

System (NWIS; data available at http://nwis.waterdata.usgs.

gov/oh/nwis/qwdata). 

Microbial Source Tracking (MST) 

Marker Analyses

Water samples for MST analyses were filtered onto 

Nucleopore polycarbonate filters (Whatman/GE Healthcare, 

Piscataway, N.J.) at the USGS Columbus laboratory within 

24 hours of sample collection. Filters were preserved at 

−70 °C in screw-cap vials with 0.3 g of acid-washed glass 

beads (Sigma, St. Louis, Mo.) until batch analysis was 

done after sample collection for the recreational season was 

completed. Samples were extracted by use of a DNA-EZ 

extraction kit (GeneRite, North Brunswick, N.J.) according to 

manufacturer’s instructions, except that no prefilter was used 

and the final elution volume was 200 μL. 

At the Columbus laboratory, DNA extracts were analyzed 

by qPCR for five MST markers: a general Bacteroides marker 

present in most warmblooded animals—AllBac (Layton and 

others, 2006); a human-associated Bacteroides marker—

HF183 (Seurinck and others, 2005); a ruminant-associated 

Bacteroides marker—BoBac (Layton and others, 2006); a 

dog-associated Bacteroides marker—BacCan (Kildare and 

others, 2007); and a gull-associated marker from the species 

Catellicoccus marimammalium—Gull2 (Sinigalliano and 

others, 2010). All qPCR assays were performed as described 

in the citations, with the exception of the Gull2, in which 

a double-quenched probe was used. The probe, which was 

labeled with an internal ZEN Quencher and an Iowa Black 

Quencher (Integrated DNA Technologies, Coralville, Iowa), 

was designed to reduce background and increase qPCR signal. 

Analyses by qPCR were run in duplicate by using 5 μL of 

DNA extract and 20 μL of master mix in an Applied Biosystems 7500 (Foster City, Calif.) thermal cycler. Depending on 
4 Microbial Source Tracking Markers at Three Inland Recreational Lakes in Ohio, 2011

the MST assay, either TaqMan Universal PCR Master Mix or 

SYBR Green PCR Master Mix was used.

Plasmid standards for each MST assay were used to 

establish standard curves for quantification of MST markers. These were constructed by insertion of PCR-amplified 

marker sequences into the pCR4 TOPO E. coli plasmid 

vector (Invitrogen, Carlsbad, Calif.). The plasmid DNA was 

extracted and purified from E. coli cells by using the QIAPrep 

Spin Miniprep Kit (Qiagen, Valencia, Calif.). Copy number 

was calculated from DNA concentration, measured by use of 

the PicoGreen assay (Invitrogen, Carlsbad, Calif.), and the 

molecular weight of the plasmid. Sample results were reported 

as copies per 100 milliliters (copies/100 mL). Standard curve 

characteristics are included in table 2.

Laboratory quality-control samples included a processing blank (phosphate-buffered water processed with the water 

samples when filtered), an extraction blank (GeneRite Elution 

Buffer processed with filtered water samples during each batch 

of DNA extraction), a no-template control (molecular-grade 

water included on each qPCR plate), and positive controls 

(plasmid-based standard curves included in each qPCR 

run). Matrix inhibition was tested by using matrix spikes, as 

described in Stoeckel and others (2011).

Table 2. Standard curve characteristics for microbial source tracking 

(MST) markers.

[Dynamic range and limit of detection are reported in copies/reaction]

MST 

marker

Number of 

compiled 

curves

Dynamic 

range

Amplification 

efficiency 

(percent)

R2

value

Limit of 

detection

AllBac 13 4.08 × 102

 – 8.72 × 107 92 0.997 8.1

BacCan 7 4.84 × 101

 – 7.70 × 106 89 0.999 42.2

BoBac 16 3.62 × 101 – 6.38 × 106 90 0.996 3.2

Gull2 2 5.52 × 101 – 5.52 × 106 82 0.997 1

HF183 19 2.26 × 101

 – 6.06 × 106 98 0.996 6.3

Table 3. Quality-control field blank data for Escherichia coli and microbial source tracking (MST) 

markers, 2011.

[Escherichia coli (E. coli) are reported in most probable number per 100 milliliters (MPN/100 mL); MST markers are 

reported in copies per 100 milliliters (copies/100 mL); <, less than; —, not done; E, estimated value]

Date Sampling site E. coli AllBac, 

General

HF183, 

Human

BoBac, 

Ruminant

BacCan, 

Dog

Gull2, 

Gull

7/31

8/14

8/30

9/6

Atwood Main 

Atwood Main 

Buckeye Crystal

Buckeye Fairfield

<1

—

<1

<1

<160

<530

<160

<530

E 400

<500

<130

<430

<64

<260

<64

<210

<840

<3,400

<840

<2,800

<20

<80

<20

<67

Escherichia coli and MST Markers in 

Lake-Water Samples

Quality-control samples for E. coli and MST markers 

were collected to aid in data interpretations. In the four field 

blanks, sample results were below detection for all but one 

sample (table 3). For HF183 (human), an estimated concentration of 400 copies/100 mL was found in one field blank 

sample. Seven concurrent replicates for E. coli and MST 

markers were collected (table 4), and the absolute value log10

differences (AVLD) were calculated for each replicate pair 

when at least one replicate included a detection. The ranges 

of AVLDs for AllBac and Gull2 markers (0.01–0.51 copies/100 mL) were similar to the range of AVLDs for E. coli
Escherichia coli and MST Markers in Lake-Water Samples 5

(0.07–0.61 MPN/100 mL). For HF183, BoBac, and BacCan 

markers, however, most AVLDs were expressed as greaterthan values because one replicate was below detection. The 

largest measured AVLD (that was not a less-than value) was 

0.87 copies/100 mL for BoBac. 

In all, 32 samples were collected for E. coli and MST 

markers—15 from Buckeye Lake sites (Crystal and Fairfield) 

and 17 from MWCD sites (Atwood and Tappan) (table 5). 

E. coli concentrations ranged from 19 to 3,000 MPN/100 mL, 

and eight samples exceeded the single-sample bathing-water 

concentration of 235 MPN/100 mL (Ohio Department of 

Health, 2011). The general marker (AllBac) was detected in all 

samples, with concentrations ranging from 1,000 to 590,000 

copies/100 mL. As found by Pearson’s correlation analysis, 

log concentrations of the general marker were not significantly 

related to log E. coli concentrations (r = 0.22, p = 0.2319). 

Concentrations of host-associated MST markers were 

placed into three categories: (1) below the detection limit for 

all markers or lowest blank value for HF183 (not detected), 

(2) detected, but value was estimated because marker was 

detected in one replicate and not the other or concentration was below the linear range (estimated quantity), and 

(3) detected and reported with certainty (measured quantity). 

Using these categories, the percentages of detections for 

human, ruminant, dog, and gull markers for Buckeye Lake, 

Atwood , and Tappan sites are shown in figure 2. The two 

Buckeye Lake sites (Crystal and Fairfield) and the two Tappan sites (Main and ditch) were combined because sources of 

fecal contamination and MST results were similar. The human 

marker was not detected at the two Buckeye Lake sites but 

was detected at Atwood and Tappan sites in 14 percent and 

50 percent of samples, respectively (estimated and measured 

quantities combined). Detections of the human marker at 

Tappan may have been from the nearby wastewater treatment plant and at Atwood and Tappan from the swimmers 

themselves. Using Pearson’s correlation analysis on data from 

the larger monitoring study, the number of swimmers was significantly related to E. coli concentrations at Atwood (r = 0.39, 

p < 0.0001) and Tappan (r = 0.20, p = 0.0187). The ruminant 

marker was detected most frequently, in 73, 57, and 60 percent 

Table 4. Quality-control concurrent replicate data for Escherichia coli and microbial source tracking (MST) markers, 2011.

[AVLD is absolute value log difference between replicate A and B; Escherichia coli (E. coli) are reported in most probable number per 100 milliliters 10 

(MPN/100 mL); MST markers are reported in copies per 100 milliliters (copies/100 mL); <, less than; >, greater than; ND, not determined because there were 

two values below detection; —, not done]

E. coli AllBac, General HF183, Human

Date Site A B AVLD A B AVLD A B AVLD

7/11

7/31

8/27

8/30

9/6

7/25

8/7

Atwood AM

Atwood PM

Atwood PM

Crystal

Fairfield

Tappan Main

Tappan Main

6

500

21

490

19

4,900

—

7

820

50

980

15

1,200

—

0.07

0.21

0.38

0.30

0.10

0.61

—

3,300

12,000

12,000

370,000

780,000

87,000

160,000

3,400

27,000

39,000

390,000

400,000

72,000

490,000

0.01

0.35

0.51

0.02

0.29

0.08

0.49

560

<130

<430

<430

<430

<220

<500

<220

300

<430

<430

<430

<220

850

>0.41

>0.36

ND

ND

ND

ND

>0.23

BoBac, Ruminant BacCan, Dog Gull2, Gull

Date Site A B AVLD A B AVLD A B AVLD

7/11

7/31

8/27

8/30

9/6

7/25

8/7

Atwood AM

Atwood PM

Atwood PM

Crystal

Fairfield

Tappan Main

Tappan Main

440

<64

<210

2,300

3,600

290

1,600

<110

<64

<210

<210

490

<110

4,400

>0.60

ND

ND

>1.04

0.87

>0.42

0.44

3,000

1,100

<2,800

<2,800

8,700

<1,400

<3,400

2,400

<840

3,700

<2,800

<2,800

<1,400

9,600

0.10

>0.12

>0.12

ND

>0.49

ND

>0.45

<33

<20

<67

<67

<67

2,300

570

<33

<20

<67

<67

<67

1,200

570

ND

ND

ND

ND

ND

0.28

0.00
6 Microbial Source Tracking Markers at Three Inland Recreational Lakes in Ohio, 2011

Table 5. Concentrations of microbial source tracking (MST) markers in inland lake samples, 2011.

[E. coli, Escherichia coli; A, average of two replicates; >, greater than; <, less than; R, value reported was detected in one field replicate but not 

in the second replicate; E, estimated value because it was detected in one replicate and not the other or was below the linear range; detections of 

AllBac, BoBac, BacCan and Gull2 are shaded] 

MST markers (copies/100 mL) 

Date Time Site E. coli

(MPN/ 100 mL)

AllBac 

(general)

HF183 

(human)

BoBac

 (ruminant)

BacCan 

(dog)

Gull2 

(gull)

7/11 9:55 Buckeye Crystal 29 130,000 <500 E 1,300 E 3,800 <80

7/17 7:05 Buckeye Crystal 58 160,000 <500 E 550 <3,400 <80

7/24 7:10 Buckeye Crystal 130 390,000 <500 E 2,200 <3,400 <80

7/30 6:55 Buckeye Crystal 920 310,000 <370 E 670 4,900 <57

8/8 8:22 Buckeye Crystal 24 160,000 <500 <260 44,000 <80

8/22 7:12 Buckeye Crystal 650 310,000 <430 <210 <2,800 <67

8/30 8:10 Buckeye Crystal A 740 A 380,000 <430 RE 2,300 <2,800 <67

9/13 10:50 Buckeye Crystal 39 330,000 <430 E 1,600 <2,800 <67

7/17 7:38 Buckeye Fairfield 19 180,000 <500 <260 5,100 <80

7/24 8:00 Buckeye Fairfield 210 200,000 <500 E 1,100 <3,400 <80

7/25 10:07 Buckeye Fairfield 210 46,000 <500 E 520 <3,400 <80

7/30 7:35 Buckeye Fairfield 80 190,000 <370 E 540 7,400 <57

8/8 8:02 Buckeye Fairfield 26 170,000 <500 <260 <3,400 <80

8/22 8:00 Buckeye Fairfield 39 180,000 <430 E 1,500 <2,800 <67

9/6 9:33 Buckeye Fairfield A 17 A 590,000 <430 AE 2,000 RE 8700 <67

7/11 9:19 Atwood Main A 6 AE 3,400 RE 560 RE 440 A 2,700 <33

7/25 9:42 Atwood Main 60 200,000 <220 E 290 <1,400 <33

7/31 8:47 Atwood Main 32 E 1,000 <130 E 270 <840 <20

8/14 9:03 Atwood Main 32 E 6,800 <500 <260 <3,400 <80

7/31 12:41 Atwood Main A 660 A 20,000 RE 300a <64 RE 1,100 <20

8/27 14:00 Atwood Main A 36 AE 26,000 <430 <210 RE 3,700 <67

9/11 13:38 Atwood Main 210 42,000 <430 E 960 <2,800 <67

7/11 10:54 Tappan Main 201 E 10,000 <220 E 190 12,000 <33

7/25 10:53 Tappan Main A 3,000 A 80,000 <220 RE 290 <1,400 A 1,800

8/7 13:17 Tappan Main 120 A 320,000 RE 850 AE 3,000 RE 9,600 AE 570

8/14 10:24 Tappan Main 96 E 9,800 1,600 <64 <840 <20

8/15 10:17 Tappan Main 21 E 51,000 <500 E 420 <3,400 <80

7/11 11:16 Tappan ditch 340 45,000 <130 <110 <1,400 <33

7/25 11:09 Tappan ditch 120 E 10,000 <220 <110 <1,400 <33

8/7 17:53 Tappan ditch 2000 590,000 E 590 <260 <3,400 <80

8/14 10:39 Tappan ditch >2400 65,000 E 710 E 930 5,100 <33

8/15 13:10 Tappan ditch ND 350,000 2,600 E 550 <3,400 <80

a

 Not considered a detection because it was below the value found in one field blank.
Escherichia coli and MST Markers in Lake-Water Samples 7

EXPLANATION

Not detected

 Estimated quantity

 Measured quantity 0

20

40

60

80

100

Human Ruminant Dog Gull

Detections, in percent

A B C A B C A B C A B C

Figure 2. Percentages of samples with not detected, estimated quantities, and measured quantities of microbial source tracking 

markers: A, Buckeye Lake Crystal and Fairfield. B, Atwood Lake. C, Tappan Lake Main and ditch, 2011.

of samples from Buckeye Lake, Atwood, and Tappan, respectively, with some uncertainty in the measurements in all samples. The dog marker was detected in 40, 43, and 30 percent 

of samples from Buckeye Lake, Atwood, and Tappan, respectively. The gull marker was found in only two samples, both 

from Tappan Main. Results from the Wilcoxon rank-sum test 

indicated that the median concentrations of E. coli in samples 

with detections of markers were not significantly different 

from samples without detections of human (p=0.24), ruminant 

(p=0.40), and dog (p=0.15) markers. This test was not done 

on the gull marker because only two detections were found.

In this study, samples were collected and analyzed by use 

of microbial source tracking techniques to take the first step 

towards understanding sources of fecal contamination at five 

sites at three inland recreational lakes. It should be emphasized, however, that microbial source tracking techniques 

are in an experimental and developmental phase as a field of 

science specialization. Experts in the field recommend that 

multiple methods be used to characterize fecal contamination 

at a site and that source feces from the study area be tested 

to confirm that source-associated MST markers are present 

(Sargeant and others, 2011). Given these limitations, in order 

to fully characterize sources at the Buckeye Lake sites, more 

work needs to be done to collect samples under a variety 

of hydrological and environmental conditions, collect fecal 

source samples in the watershed, collect afternoon samples 

when swimmers are present, and apply other source tracking 

tools to establish multiple lines of evidence.
8 Microbial Source Tracking Markers at Three Inland Recreational Lakes in Ohio, 2011

References Cited

Bower, P.A., Scopel, C.O., Jensen, E.T., Depas, M.M., and 

McLellan, S.L., 2005, Detection of genetic markers of fecal 

indicator bacteria in Lake Michigan and determination of 

their relationship to Escherichia coli densities using standard microbiological methods: Applied and Environmental 

Microbiology, v. 71, no. 12, p. 8305–8313. 

Edge, T.A., Hill, S., Seto. P., and Marsalek, J., 2010, Librarydependent and library-independent microbial source 

tracking to identify spatial variation in faecal contamination sources along a Lake Ontario beach: Water Science & 

Technology, v. 62, no. 3, p. 719–727. 

Kildare, B.J., Leutenegger, C.M., McSwain, B.S., Bambic, 

D.G., Rajal, V.B., and Wuertz, S., 2007, 16S rRNA-based 

assays for quantitative detection of universal, human-, 

cow-, and dog-specific fecal Bacteroidales—A Bayesian 

approach: Water Research, v. 41, no. 16, p. 3701–3715.

Layton, A., McKay, L., Williams, D., Garrett, V., Gentry, R., 

and Sayler, G., 2006, Development of Bacteroides 16S 

rRNA gene TaqMan-based real-time PCR assays for estimation of total, human, and bovine fecal pollution in water: 

Applied and Environmental Microbiology, v. 72, no. 6, 

p. 4214–4224.

Myers, D.N., Stoeckel, D.M., Bushon, R.N., Francy, D.S., and 

Brady, A.M.G., 2007, Fecal indicator bacteria (ver. 2.0): 

U.S. Geological Survey Techniques of Water-Resources 

Investigations, book 9, chap. A7, section 7.1, accessed 

October 2011 at http://pubs.water.usgs.gov/twri9A/.

Ohio Department of Health, 2011, Bathing beach monitoring 

program, accessed October 2011 at http://www.odh.ohio.

gov/odhprograms/eh/bbeach/beachmon.aspx.

Seurinck, S., Defoirdt, T., Verstraete, W., and Siciliano, S.D., 

2005, Detection and quantification of the human-specific 

HF183 Bacteroides 16S rRNA genetic marker with realtime PCR for assessment of human faecal pollution in freshwater: Environmental Microbiology, v. 7, no. 2, p. 249–259.

Sinigalliano, C.D., Fleisher, J.M., Gidley, M.L., Solo-Gabriele, 

H.M., Shibata, T., Plano, L.R., Elmir, S.M., Wanless, D., 

Bartkowiak, J., Boiteau, R., Withum, K., Abdelzaher, 

A.M., He, G., Ortega, C., Zhu, X., Wright, M.E., Kish, J., 

Hollenbeck, J., Scott, T., Backer, L.C., and Fleming, L.E., 

2010, Traditional and molecular analyses for fecal indicator 

bacteria in non-point source subtropical recreational marine 

waters: Water Research, v. 44, no.13, p. 3763–3772.

Sargeant, D., Kamin, W.R., and Collyard, S., 2011, Review 

and critique of current microbial source tracking (MST) 

techniques: Olympia, Wash., Washington State Department of Ecology Publication No. 11-03-038, accessed 

September 2012 at https://fortress.wa.gov/ecy/publications/

SummaryPages/1103038.html, 66 p.

Stoeckel, D.M., Stelzer, E.A., Stogner, R.W., and Mau, D.P., 

2011, Semi-quantitative evaluation of fecal contamination potential by human and ruminant sources using 

multiple lines of evidence: Water Research, v. 45, no. 10, 

p. 3225–3244.

Wong, M., Kumar, L., Jenkins, T.M., Xagoraraki, I., Phanikumar, M.S., and Rose, J.B., 2009, Evaluation of public health 

risks at recreational beaches in Lake Michigan via detection of enteric viruses and a human-specific bacteriological 

marker: Water Research, v. 43, no. 4, p. 1137–1149.

Francy and Stelzer—Microbial Source Tracking Markers at Three Inland Recreational Lakes in Ohio, 2011—Open-File Report 2012–1222