occurrence and distribution of fecal indicator …occurrence and distribution of fecal indicator...

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

Open-File Report 2015–1013

Prepared in cooperation with the Great Lakes Restoration Initiative

Occurrence and Distribution of Fecal Indicator Bacteria and Gene Markers of Pathogenic Bacteria in Great Lakes Tributaries, March–October 2011

Cover: Bad River, Wisconsin (photo by Eric Dantoin, U.S. Geological Survey).


By Angela K. Brennan, Heather E. Johnson, Alex R. Totten, and Joseph W. Duris

Prepared in cooperation with the Great Lakes Restoration Initiative

Open-File Report 2015–1013

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

U.S. Department of the InteriorSALLY JEWELL, Secretary

U.S. Geological SurveySuzette M. Kimball, Acting Director

U.S. Geological Survey, Reston, Virginia: 2015

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Suggested citation:Brennan, A.K., Johnson, H.E., Totten, A.R., and Duris, J.W., 2015, Occurrence and distribution of fecal indicator bacte-ria and gene markers of pathogenic bacteria in Great Lakes tributaries, March–October 2011: U.S. Geological Survey Open-File Report 2015–1013, 29 p., http://dx.doi.org/10.3133/ofr20151013.

ISSN 2331-1258 (online)

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http://www.usgs.gov/pubprod

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iii

Contents

Abstract ...........................................................................................................................................................1Introduction.....................................................................................................................................................1

Purpose and Scope ..............................................................................................................................2Description of Study Area ...................................................................................................................2

Sampling and Analytical Methods ..............................................................................................................2Site Selection.........................................................................................................................................2Sample Collection .................................................................................................................................2Microbiological Analyses ...................................................................................................................6

Bacteria Enumeration, Enrichment, and Preservation ..........................................................6DNA Extraction .............................................................................................................................6Polymerase Chain Reaction Assays for Pathogen Detection ..............................................6

Results of Microbiological Analyses ..........................................................................................................6Occurrence and Distribution of Fecal Indicator Bacteria .............................................................6Occurrence and Distribution of Bacterial Pathogen Gene Markers ...........................................8

Quality-Control Results ...............................................................................................................................10Summary........................................................................................................................................................10Acknowledgements .....................................................................................................................................11References Cited..........................................................................................................................................11Appendixes ................................................................................................................................................13

Figures 1. Graph showing land cover surrounding tributary sites sampled as part of the

Great Lakes Restoration Initiative (GLRI) pathogens study ...................................................3 2. Map showing Great Lakes Restoration Initiative (GLRI) USGS tributary sampling

locations and number of samples collected for pathogens study, 2011 .............................4 3. Graph showing frequency of samples testing positive for pathogen gene markers ........8 4. Graph showing percentage of pathogen gene markers in relation to water quality

criteria.....................................................................................................................................................9

Tables 1. Great Lakes Restoration Initiative (GLRI) pathogens study sampling locations, 2011 ......5 2. Growth media, target organisms, and incubation temperature requirements for the

Great Lakes Restoration Initiative (GLRI) pathogens study ...................................................7 3. Selected pathogen gene targets and virulence traits of the organism ...............................7 4. Summary of the frequency of pathogen gene detections in a single sample. ...................8 5. Percentage of samples that tested positive for pathogenic gene markers during

high-flow and normal-flow conditions ......................................................................................9 6. Relative standard deviation of field replicates for quantitative microbial methods

in the Great Lakes Restoration Initiative (GLRI) pathogens study ......................................10 7. Frequency of agreement between field replicates for qualitative microbial

methods in the Great Lakes Restoration Initiative (GLRI) pathogens study .....................10

iv

Multiply By To obtain

Length

mile (mi) 1.609 kilometer (km)Area

square mile (mi2) 2.590 square kilometer (km2) Volume

milliliter (mL) 1,000 microliter (µL)

Temperature in degrees Celsius (°C) may be converted to degrees Fahrenheit (°F) as follows: °F=(1.8×°C)+32Horizontal coordinate information is referenced to North American Datum of 1983 (NAD 83).Concentrations of fecal indicator bacteria are given in colony-forming units per 100 milliliters (CFU/100 mL).

Conversion Factors

v

Abbreviations used in report

CDC Centers for Disease Control and Prevention

CFU colony-forming unit

DNA Deoxyribonucleic acid

E. coli Escherichia coli

EWI equal-width-increment

FIB Fecal indicator bacteria

GLRI Great Lakes Restoration Initiative

MI-BaRL USGS Michigan Bacteriological Research Laboratory

MI-WSC USGS Michigan Water Science Center

PBS phosphate buffered saline

PCR polymerase chain reaction

QC quality control

RSD relative standard deviation

STEC Shiga-toxin producing Escherichia coli

EPA U.S. Environmental Protection Agency

USGS U.S. Geological Survey

WQC water quality criteria


By Angela K. Brennan, Heather E. Johnson, Alex R. Totten, and Joseph W. Duris

AbstractFrom March through October 2011, the U.S. Geological

Survey (USGS), conducted a study to determine the frequency of occurrence of pathogen gene markers and densities of fecal indicator bacteria (FIB) in 22 tributaries to the Great Lakes. This project was funded as part of the Great Lakes Restora-tion Initiative (GLRI) and included sampling at 22 locations throughout 6 states that border the Great Lakes.

A total of 177 environmental samples were collected at USGS streamgaging stations during both normal-flow and high-flow conditions and were analyzed by the Michigan Bac-teriological Research Laboratory at the USGS Water Science Center in Lansing, Michigan.

Water samples were analyzed for the presence of FIB concentrations (FIB; fecal coliform bacteria, Escherichia coli [E. coli], and enterococci) by using membrane filtration and serial dilution methods. The resulting enrichments from standard culturing of the samples were then analyzed by using polymerase chain reaction (PCR) to determine the occurrence of pathogen gene markers for Shigella species, Campylobacter jejuni and coli, Salmonella species, and pathogenic E. coli, including Shiga toxin-producing E. coli (STEC).

IntroductionIn 2009, the Great Lakes Restoration Initiative (GLRI)

was started as an interagency initiative to protect and restore the Great Lakes. This report describes data collected in 2011 as part of the U.S. Geological Survey (USGS) GLRI effort led by the Michigan Water Science Center (MI-WSC), in conjunc-tion with USGS Water Science Centers in Indiana, Minnesota, Ohio, New York, and Wisconsin, and complements the ongo-ing USGS GLRI Great Lakes Nutrient and Sediment Loadings study (GLRI Loadings study) being conducted at the same sampling locations.

The microbiological monitoring network for this study consisted of 22 sampling locations, representing multiple land-use types, drainage areas, and soil characteristics typical of the Great Lakes region. Samples were collected to better understand the occurrence and distribution of fecal indicator bacteria (FIB) and selected bacterial pathogens that are com-monly associated with waterborne illnesses.

Within the broad FIB groups (Escherichia coli [E. coli], fecal coliforms, and enterococci), there are specific strains of bacteria that are pathogenic (infection causing) to humans. The bacterial pathogen gene markers evaluated in this study included genes from the genera Shigella, Campylobacter (jejuni and coli) and Salmonella. In addition, several genes were also evaluated from a class of pathogenic E. coli known as Shiga toxin-producing E. coli (STEC), which includes E. coli O157:H7.

Fecally derived bacterial pathogens have long been known to be a health threat to humans, and exposure to these pathogens can result in illness and occasionally even death (Centers for Disease Control and Prevention [CDC], 2012b). STEC, including E. coli O157:H7, can cause illness ranging from mild intestinal disease to severe kidney complications and death in animals and humans (CDC, 2012d). Shigella’s pathogenicity functions similarly to that of STEC; however, Shigella mainly affects humans. Campylobacter is one of the most common causes of diarrheal illness in the United States, and symptoms include cramping, abdominal pain, and fever (CDC, 2012a). Salmonella infection can cause diarrhea, fever, and abdominal cramps, and severe cases can even lead to death (CDC, 2012c).

Current methods for evaluating recreational and drinking water safety typically rely on the enumeration of FIB (U.S. Environmental Protection Agency [EPA], 2012). FIB are used to indicate the potential for illness associated with fecal contamination (EPA, 2012). Primary contact refers to water-contact activities where immersion and ingestion are likely, such as swimming, water skiing, and diving. Even though most tributaries are not used for “primary contact” recreational

2 Fecal Indicator Bacteria and Gene Markers of Pathogenic Bacteria in Great Lakes Tributaries

water activities, many of these major tributaries enter the Great Lakes at or near a beach which is designated as a recre-ation area. Most tributaries are used for partial, or secondary, contact recreation, indicating water activities where there is the potential of submergence, such as boating and fishing. FIB concentrations and the occurrence of bacterial pathogens associated with these bodies of water are important, not only to partial contact recreational users in rivers but ultimately to primary contact recreational users at Great Lakes beaches. The partial body contact recreation standard (Dufour and Bal-lantine, 1986) was used for data interpretation for this report because the EPA 2012 guidance does not include a numerical standard for partial body contact in recreational waters.

Purpose and Scope

This report presents data obtained during a Great Lakes-wide study conducted in 2011 by the USGS as part of the GLRI. The purpose of this study was to determine the occur-rence and distribution of FIB and pathogenic gene markers within 22 major tributaries to the Great Lakes. Normal-flow and high-flow samples were analyzed for FIB and for patho-gens Shigella, Campylobacter (jejuni and coli), Salmonella, and STEC, which included E. coli O157:H7. The relations of FIB concentrations and bacterial pathogen gene occurrence to recreational water quality criteria were also assessed.

Description of Study Area

Approximately 132,208 miles of tributaries (EPA and USGS, 2012), with a drainage area of almost 295,000 square miles (Great Lakes Information Network, 2013), drain into the Great Lakes from six states (Indiana, Michigan, Minnesota, New York, Ohio, Wisconsin) and Canada. The 22 monitoring locations are grouped on the basis of predominant land cover surrounding the major tributaries in this study (fig. 1) (USGS, 2011) and were distributed geographically throughout the Great Lakes region of the United States (fig. 2).

Sampling and Analytical Methods

Site Selection

All sampling locations were at USGS streamgaging stations. Twenty-two sampling locations (fig. 2, table 1) were selected (out of 59 GLRI Loadings study sites) to create a geo-graphically distributed study and to address the variability in land cover throughout the Great Lakes region. Sampling loca-tions, as well as timing of sample collection, were designed to complement the GLRI Loadings study (USGS, 2014). In addi-tion to those sites selected from the GLRI Loadings study, the Paw Paw River at Riverside, MI, was included as a sampling location for this study because the site was routinely visited as part of another USGS MI-WSC project.

Sample Collection

Hydrologic conditions were determined on the basis of mean daily discharge1 at each USGS streamgage on the date of collection, calculated from date of first record to current. Con-ditions were evaluated such that if daily discharge2 was greater than or equal to the 75th percentile, the corresponding sample was considered a high-flow sample; otherwise, the sample was considered a normal-flow sample. Owing to a short period of record, the Grand River at Grand Rapids station was used to estimate hydrologic conditions for the Grand River at Eastmanville station. Water samples were collected during normal-flow conditions from March through October 2011 in an effort to target the recreational water season. Samples were also collected during high-flow conditions (storm events), but sampling frequency varied among sites in response to the frequency of storm events and availability of staff (table 1).

The sampling frequency depended on the sampling schedule of the GLRI Loadings study. Some sites were sampled more often than others, including those sites targeted for analysis of certain chemical constituents (such as waste-water indicator compounds) in addition to FIB and pathogenic gene markers.

Normal-flow and high-flow sampling events typically coincided with GLRI Loadings study sampling. All samples were collected by USGS personnel using either equal-width-increment (EWI) or multiple vertical sampling techniques designed to account for the variable distribution of constitu-ents across the stream channel and throughout the water column (USGS, 2006). EWI samples were typically collected off bridges by using a crane apparatus with an attached DH-95 depth integrating sampler, or from boats. There were instances when wading measurements were made, but typically only during normal-flow conditions.

For each sample, a 1-liter sterile high density polyethyl-ene (HDPE) sample bottle was filled to the shoulder to allow some head space for sample mixing. Once collected, water samples were placed on ice in a cooler (away from ultraviolet light) and shipped overnight to the MI-BaRL (Michigan Bac-teriological Research Laboratory) at the MI-WSC in Lansing, Michigan. Upon receipt of the shipments, sample information and streamflow condition were logged in to the laboratory database system, and samples were placed into the refrigerator at 4–7 degrees Celsius (°C) until processing and analysis (typi-cally the same day).

A total of 177 water samples were collected, including field quality-control (QC) samples. There were 59 normal-flow samples and 54 high-flow samples used for data analysis. Quality-control samples included 21 field blank samples, 21 source solution blank samples, and 22 field replicate samples.

1The arithmetic mean of discharges for all instances of a particular calendar day on record or for a specific set of years.

2The arithmetic mean discharge for a single day.

Sampling and Analytical Methods 3

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Sampling and Analytical Methods 5

Field blanks, source solution blanks, and field replicate sam-ples were collected according to the methods described in the USGS National Field Manual, with inorganic-grade reagent water being used as the sample medium (USGS, 2006). Twelve samples were excluded from the data analysis because the samples exceeded the holding time (48 hours) established

by the MI-BaRL for FIB culturing for nonregulatory purposes. There were also three instances where the bacteria plates were uncountable (coded as “unc”), owing to smearing or a heavy amount of particulates on the filter. A total of 165 water samples were included in the final analysis.

Table 1. Great Lakes Restoration Initiative (GLRI) pathogens study sampling locations, 2011.

[nr, near]

USGS site nameUSGS site

identification number

Field ID

Number of samples collected during

normal flow

Number of samples collected during

high flow

Indiana

Burns Ditch at Portage 04095090 BN 3 4MichiganAu Sable River nr Au Sable 04137500 AU 5 1Clinton River at Moravian Drive at Mt. Clemens 04165500 CL 0 2Grand River nr Eastmanville 04119400 GR 0 5Ontonagon River nr Rockland 04040000 ON 8 1Paw Paw River at Riverside 04102500 PP 2 5River Rouge at Detroit 04166500 RR 1 2River Raisin nr Monroe 04176500 RS 2 0Saginaw River at Saginaw 04157000 SG 5 3St. Joseph River at Niles 04101500 SJ 3 4

Minnesota

St. Louis River at Scanlon 04024000 ST 7 2New York

Genessee River at Ford St. Bridge at Rochester 04231600 GS 3 3Oswego River at Lock 7 at Oswego 04249000 OS 4 4St. Regis River at Brasher Center 04269000 SR 5 5

Ohio

Cuyahoga River at Independence 04208000 CY 1 4Maumee River at Waterville 04193500 MA 2 1Portage River at Woodville 04195500 PO 0 3

Wisconsin

Bad River nr Odanah 04027000 BD 3 1Fox River at Oil Tank Depot at Green Bay 040851385 FX 1 1Menominee River nr McAllister 04067500 MM 2 0Manitowoc River at Manitowoc 04085427 MN 1 1Milwaukee River at Milwaukee 04087170 MW 1 1


Microbiological Analyses

Bacteria Enumeration, Enrichment, and Preservation

Water samples were analyzed for the presence of FIB and FIB concentrations by using standard membrane filtration and serial dilution methods (Myers and others, 2014). Fecal coli-form bacteria were analyzed according to standard methods of the American Public Health Association and others (1998), E. coli bacteria were analyzed according to EPA method 1603 (EPA, 2009a), and enterococci bacteria were analyzed accord-ing to EPA method 1600 (EPA, 2009b). Media were prepared according to manufacturer’s instructions.

A volume of 50 milliliters (mL) of sample water was filtered through a 0.45-micrometer-pore-size nylon-membrane filter (Advantec, USA) by means of aseptic techniques. The enriched filter was then placed on a selective growth medium for enumeration and enrichment of target bacterial pathogen groups and incubated at the optimal temperature (table 2).

The filters with E. coli, enterococci, and fecal coliform bacteria growth were transferred to a vial containing 1 mL of phosphate buffered saline solution (PBS), preserved with 20 percent glycerol, and frozen at −70 °C until further analysis; these samples are referred to as “glycerol stocks.” The filters with Campylobacter and Salmonella growth were aseptically removed from the Gram Negative and Bolton Broth enrich-ments, and the remaining culture was centrifuged to form a pellet. The supernatant was decanted, and the pellet was resuspended in 1 mL of PBS solution and preserved with 20 percent glycerol and frozen at −70 °C until deoxyribonucleic acid (DNA) extraction.

DNA ExtractionGlycerol stocks were thawed at room temperature and

DNA was extracted by using the Qiagen QIAamp® DNA Mini Kit (Qiagen, USA), following the manufacturers proto-col. DNA concentrations and purity were determined with a NanoDrop spectrophotometer (Thermo Scientific, USA). DNA was then stored at −20 °C until needed for Polymerase Chain Reaction (PCR) analysis for the selected gene targets (table 3).

Polymerase Chain Reaction Assays for Pathogen Detection

Samples were analyzed by using PCR to determine the occurrence of pathogen gene markers for Shigella sonnei, Campylobacter jejuni and coli, Salmonella, and STEC. The primer sequences, positive controls, reaction details, and qual-ity-control procedures used for the detection of the pathogens by PCR listed in table 3 were based on previously published studies (Haack and others, 2013).

All PCR assays were performed by using 1 microliter of DNA solution, representing 0.1–85 nanograms of template DNA, which was isolated from the appropriate growth enrich-ment. Results from these analyses indicated whether the target genes were present or absent in the sample above the method detection limit; however, PCR does not quantify the genes present. In order for the gene to be detected, the organism had to grow on the appropriate medium, and therefore must have been viable in the sample.

Laboratory filtration blanks were processed daily with every set of samples on all growth media and enrichments to ensure that the laboratory equipment was sterile and that aseptic laboratory techniques were being followed. One set of laboratory replicates was processed weekly on all growth media and enrichments to quantify method variability. If con-tamination was found in the laboratory blanks, either sample results were discarded or samples were rerun.

Standard quality-assurance and quality-control pro-cedures were followed for all PCR reactions (EPA, 2004). Detection limits for all PCR reactions were determined by using serial dilution of DNA from control organisms. For approximately every 20 samples of any given PCR assay, PCR positive controls (DNA extracted from bacteria known to con-tain the target gene) and PCR negative controls (no template reactions) were included.

Results of Microbiological AnalysesOf the 177 water samples collected, 12 samples were

omitted from the data analysis because of holding-time viola-tions and 64 of the samples were collected for the analysis of quality control. The 165 samples included in the analyses were assessed for occurrence and distribution of FIB during normal and high flows and for whether the samples exceeded the water quality criteria. These samples were also analyzed for specific pathogen genes by using PCR. Fecal indicator bacteria results are included in the USGS National Water Information System (NWIS) database.

Occurrence and Distribution of Fecal Indicator Bacteria

Fecal coliform concentrations ranged from < 2 to 950 colony-forming units per 100 mL (CFU/100 mL), E. coli concentrations ranged from < 2 to 26,000 CFU/100 mL, and enterococci concentrations ranged from < 2 to 31,000 CFU/100 mL. Fecal indicator bacteria concentrations for all samples are presented in appendix 1.

The lowest E. coli concentrations were recorded at the Au Sable River near Au Sable, MI (AU) (during high and normal flows), and the St. Joseph River at Niles, MI (during normal flow) (SJ). The highest E. coli concentrations were recorded at

Results of Microbiological Analyses 7

the River Rouge at Detroit, MI (during high flow) (RR). The lowest enterococci concentrations were recorded at the Au Sable River near Au Sable, MI (AU) (during high and normal flows), and the Oswego River at Lock 7 at Oswego, NY (OS) (during high flow). The highest enterococci concentrations were recorded at the River Rouge at Detroit, MI (RR) (during high flow). The lowest E. coli and enterococci concentrations were found at sites dominated by forest and agricultural land cover, whereas the highest E. coli and enterococci concentra-tions were recorded at sites dominated by urban development.

Enterococci and E. coli concentrations were evaluated on the basis of whether the data met or exceeded the 1986 EPA criterion for moderate full body contact recreation for freshwater for a single sample (Dufour and Ballantine, 1986). The moderate full body contact recreation criterion was

considered most applicable for this study given the type of recreation typically found on rivers; for example, kayaking, canoeing, and fishing. The moderate full body contact criterion for enterococci for a single sample is greater than or equal to 78 CFU/100 mL; and for E. coli, the recreation criterion is greater than or equal to 298 CFU/100 mL for a single sample. Of 112 samples analyzed for FIB concentrations, 21 percent of samples exceeded the EPA recreation criterion for E. coli, and 40 percent of samples exceeded the criterion for enterococci.

During high-flow events, 43 percent of all samples exceeded the EPA recreation criterion for E. coli, and 61 per-cent of samples exceeded the criterion for enterococci. During normal-flow conditions, about 2 percent of samples exceeded the criterion for E. coli, and about 21 percent of samples exceeded the criterion for enterococci.

Table 2. Growth media, target organisms, and incubation temperature requirements for the Great Lakes Restoration Initiative (GLRI) pathogens study.

[mL, milliliters; °C, degrees Celsius; E. coli, Escherichia coli]

Growth media Organism or methodVolume

filtered (mL)Incubation temperature and time

requirement

mFC Fecal coliform bacteria (E. coli, Shigella)

50 44.5 °C ± 0.5 °C for 22–24 hours

Modified mTEC E. coli 50, 10, 1 35–37 °C for 2 hours, then 44.5 °C ± 0.5 °C for 20–22 hours

mEI Enterococci 50, 10, 1 41 °C ± 0.5 °C for 22–24 hoursBolton Broth (BB) with Preston1

supplementCampylobacter 50 37 °C for 4 hours, then 42 °C

for 40–44 hoursGram Negative Broth (GN) Salmonella 50 37°C ± 1.0 °C for 22–24 hours

1Oxoid, Cambridge, United Kingdom.

Pathogen Gene target Virulence trait

E. coli eaeA AttachmentE. coli stx2 Severe toxinE. coli stx1 Moderate toxinE. coli rfbO157 Common outbreak strainShigella ipaH InvasionSalmonella invA InvasionSalmonella spvC Multiple virulence traitsCampylobacter jejuni and coli 16s rDNA (Campy) Pathogenic species marker

Table 3. Selected pathogen gene targets and virulence traits of the organism.

[E. coli, Escherichia coli]


Occurrence and Distribution of Bacterial Pathogen Gene Markers

Data from the E. coli pathogen analysis showed that 68 percent of samples contained the eaeA gene, 20 percent of samples contained the stx2 gene, 12 percent of samples con-tained the stx1 gene, and 11 percent of samples contained the rfbO157 gene (fig. 3). Data from the Shigella analysis showed that 6 percent of samples contained the ipaH gene. Data from the Salmonella analysis showed that 7 percent of samples con-tained the invA gene and 14 percent of samples contained the spvC gene. Data from the Campylobacter analysis showed that 9 percent of samples contained the 16s rDNA gene (fig. 3).

Detections of bacterial pathogen gene markers by site and by sample are listed in appendix 3.

The frequency of pathogen gene markers detected in each sample is summarized in table 4. The greatest number of pathogen gene markers detected in any one sample was seven (River Rouge at Detroit, MI, during a May 26, 2011, high-flow event). This same May 26, 2011 sample from the River Rouge also contained the highest E. coli and enterococci concentra-tions of samples collected during this project (E. coli value of 26,000 CFU/100 mL; enterococci value, 31,000 CFU/100 mL).

0

10

20

30

40

50

60

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80

eaeA stx2 stx1 rfb0157 ipaH invA spvC C. coli/C. jejuni

Posi

tive

sam

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Figure 3. Frequency of samples testing positive for pathogen gene markers (C. coli, Campylobacter coli; C. jejuni, Campylobacter jejuni).

Figure 3. Frequency of samples testing positive for pathogen gene markers (C. coli, Campylobacter coli; C. jejuni, Campylobacter jejuni).

Table 4. Summary of the frequency of pathogen gene detections in a single sample.

[<, less than]

Flow conditions

Number of bacterial pathogen genes present

0 1 2 3 4 5 6 7 8

Percentage of samples in which the indicated number of genes were detected

High flow 10 12 13 2 3.5 6 0 1 0Normal flow 16 25 10 1 <1 0 0 0 0Total percent detection 26 37 23 3 4 6 0 1 0

Results of Microbiological Analyses 9

For those samples in which 3 or more genes were pres-ent, 14 of the 16 samples were collected during high-flow conditions.

Pathogen frequency was analyzed in relation to hydro-logic condition among all sites, as illustrated in figure 4. Overall, a higher percentage of samples tested positive for pathogens under high-flow conditions than during normal-flow conditions. For Campylobacter, the differences in the percent-age of samples that were positive for pathogens between high- and normal-flow conditions were less obvious. The invA gene for Salmonella was found only during high flows.

The frequencies of pathogen gene occurrence in samples that met and exceeded the EPA criteria for moderate full body contact recreation for E. coli and enterococci were compared for the 22 sites (table 5). Fisher’s Exact Test was run to deter-mine if there was a significant difference (p-value less than 0.05) between gene detection in samples that met or exceeded the water quality criteria. Overall, there was a greater occur-rence of pathogen gene markers in samples that exceeded the moderate full body contact recreation criteria for both E. coli and enterococci, the exception being a greater occurrence of the ipaH gene marker for Shigella in samples that met the criterion for E. coli.

0

10

20

30

40

50

60

70

80


Perc

ent a

ge o

f sam

ples

Gene Marker

Percentage of normal-flow samples

Percentage of high-flow samples

Figure 4. Percentage of pathogen gene markers in relation to water quality criteria (C. coli, Campylobacter coli; C. jejuni, Campylobacter jejuni).

Table 5. Percentage of samples that tested positive for pathogenic gene markers during high-flow and normal-flow conditions.

[C. coli, Campylobacter coli; C. jejuni, Campylobacter jejuni, E. coli, Escherichia coli; n, number of samples; <, less than; nsd, no significant difference; p-value, p-value less than 0.05 = significant difference between groups, Fisher’s Exact Test]

Water quality criteria1 status

n eaeA stx2 stx1 rfb0157 ipaH invA spvCC. coli/C.

jejuni

Meet E. coli criterion 88 60 11 3 5 7 0 9 6Exceed E. coli criterion 24 96 50 42 33 4 33 33 21p-value — <0.05 <0.05 <0.05 <0.05 nsd <0.05 <0.05 <0.05

Meet enterococci criterion

67 54 10 5 3 6 0 3 2

Exceed enterococci criterion

45 91 33 22 22 7 18 31 20

p-value — <0.05 <0.05 <0.05 <0.05 nsd <0.05 <0.05 <0.051Water quality criteria = EPA Water Quality Criteria for Moderate Full Body Contact (Dufour and Ballantine, 1986).


Quality-Control ResultsTo assess for bias and precision, 21 field blanks, 21

source solution blanks, and 22 field replicates were collected and processed in the same manner as the regular field samples. All field blanks and source solution water blanks were nega-tive (no growth) for all FIB except for a sample collected at the River Raisin on August 30, 2011, which had an entero-cocci count of 2E (appendix 2). For PCR analysis, all no-tem-plate controls (negative controls) were found to be negative, and all positive assay controls yielded appropriate responses. However, the eaeA, rfb0157, and ipaH genes were infre-quently detected in the field blanks and source solution blanks, most likely as a result of contamination in the source solution blank water (appendix 4). There were two dates—Septem-ber 7, 2011, and August 24, 2011—for which environmental samples were collected in association with the source solution blank and field blank samples where there may have been possible contamination. The environmental sample, source solution blank, and field blank samples from these dates have been qualified with a “V” (analyte was detected in both the environmental sample and the associated blanks) as a result of possible contamination and data were not included in analyses (appendixes 3 and 4).

For all FIB field replicates, a relative standard deviation (RSD) was calculated and a mean RSD was computed (table 6) by using the formula below:

In all cases, the average RSD fell below the recom-mended 36 percent according to the initial and ongoing precision and recovery acceptance criteria that were used as a benchmark for replicate environmental samples (table 6) (EPA, 2009a).

For pathogen analysis, which yields only a presence or absence result, the frequencies of agreement between repli-cates were determined by dividing the number of paired repli-cates in agreement by the total number of replicates (table 7).

The field blank and source solution blank data results are presented in appendixes 2 and 4. All replicate data are presented in appendixes 1 and 3, along with the associated environmental sample.

SummaryA total of 177 environmental samples were collected at

USGS streamgaging stations during both normal-flow and high-flow conditions in 22 tributaries to the Great Lakes and were analyzed for the presence of FIB concentrations and to determine the occurrence of pathogen gene markers for Shigella species, Campylobacter jejuni and coli, Salmonella species, and pathogenic E. coli, including STEC. Results from a total of 165 samples were evaluated because 12 samples did not meet quality-control measures.

E. coli concentrations ranged from less than 2 to 26,000 CFU/100 mL. Enterococci concentrations ranged from less than 2 to 31,000 CFU/100 mL. Fecal coliform bacteria concentrations ranged from less than 2 to 950 CFU/100 ml. Data from E. coli pathogen analyses showed that 68 percent of samples contained the eaeA gene, 20 percent of samples contained the stx2 gene, 12 percent of samples contained the stx1 gene, and 11 percent of samples contained the rfbO157 gene. Data from the Shigella analysis showed that 6 percent of samples contained the ipaH gene. Data from the Salmonella analysis showed that 7 percent of samples contained the invA gene and 14 percent of samples contained the spvC gene. Data from the Campylobacter analysis showed that 9 percent of samples contained the 16S rDNA gene of Campylobacter coli and Campylobacter jejuni.

Table 6. Relative standard deviation of field replicates for quantitative microbial methods in the Great Lakes Restoration Initiative (GLRI) pathogens study.

[E. coli, Escherichia coli; RSD, relative standard deviation]

Statistic E. coli EnterococciFecal

coliforms

Average RSD, in percent 25.3 28.4 31.1Number of field replicates 22 22 22

Table 7. Frequency of agreement between field replicates for qualitative microbial methods in the Great Lakes Restoration Initiative (GLRI) pathogens study.

[C. coli, Campylobacter coli; C. jejuni, Campylobacter jejuni]

StatisticPathogen gene markers


Percent agreement 82 95 95 82 100 95 91 95Number of field replicates 22 22 22 22 22 22 22 22

SX X X X X X

n=

− + − + − +

−

( ) ( ) ( ) ...1 2 3

1

2 2 2

References Cited 11

There were no samples in which all eight pathogen gene markers were detected; however, 88 percent of the samples that had three or more pathogen gene markers present were collected during high-flow conditions. Overall, there were a higher percentage of samples that tested positive for pathogens during high-flow conditions than normal-flow conditions, and there was a greater occurrence of pathogen gene markers in samples that exceeded the EPA moderate full body contact recreation criteria for E. coli and enterococci (Dufour, 1986) than in samples that met the EPA criteria; the exception was a greater occurrence of the ipaH gene marker for Shigella in samples that met the EPA criterion for E. coli than in samples that exceeded the EPA criterion for E. coli.

AcknowledgementsThis study was funded by the Great Lakes Restoration

Initiative. We wish to acknowledge the many USGS personnel from Indiana, Michigan, Minnesota, Ohio, New York and Wis-consin who collected the water samples for this project and communicated with the Michigan Bacteriological Research Laboratory.

References Cited

American Public Health Association, American Water Works Association, and Water Environment Federation, 1998, Standard methods for the examination of water and waste-water (20th ed.): Washington, D.C., American Public Health Association, p. 9–1 to 9–76.

Centers for Disease Control and Prevention (CDC), 2012a, Campylobacter: CDC Web page, accessed April 17, 2012, at http://www.cdc.gov/nczved/dfbmd/diseases/campylobacter/.

Centers for Disease Control and Prevention (CDC), 2012b, Recreational Water Illnesses (RWIs): CDC Web page, accessed April 17, 2012, at http://www.cdc.gov/healthywa-ter/swimming/rwi/.

Centers for Disease Control and Prevention (CDC), 2012c, Salmonella: CDC Web page, accessed April 17, 2012, at http://www.cdc.gov/salmonella.

Centers for Disease Control and Prevention (CDC), 2012d, STEC: CDC Web page, accessed April 17, 2012, at http://www.cdc.gov/ecoli/index.html.

Dufour, A.P., and Ballantine, R.K., 1986, Ambient water quality criteria for bacteria, 1986—Bacteriological ambient water quality criteria for marine and freshwater recreational waters: Washington, D.C., EPA 440/5–84–002.

Great Lakes Information Network (GLIN), 2013, Great Lakes facts and figures: GLIN Web page, accessed May 6, 2013, at http://www.glin.net/lakes/ref/lakefact.html.

Haack, S.K., Fogarty, L.R., Stelzer, E.A., Fuller, L.M, Bren-nan, A.K, Isaacs, N.M., and Johnson, H.E. 2013, Geo-graphic setting influences great lakes beach microbiological water quality: Environmental Science & Technology, v. 47, no. 21, p. 12054–12063.

Myers, D.N., Stoeckel, D.M., Bushon, R.N., Francy, D.S., and Brady, A.M.G., 2014, Fecal indicator bacteria (ver. 2.1): U.S. Geological Survey Techniques of Water-Resources Investigations, book 9, chap. A7, section 7.1, May 2014, accessed June 2014 at http://pubs.water.usgs.gov/twri9A/.

U.S. Environmental Protection Agency (EPA), 2004, Quality assurance/quality control guidance for laboratories per-forming PCR analysis on environmental samples: Office of Water, EPA 815-B-04-001, 56 p.

U.S. Environmental Protection Agency (EPA), 2009a, Method 1603 “Escherichia coli in water by membrane filtration using modified membrane-thermotolerant Escherichia coli agar”: Washington, D.C., EPA–821–R–09–007 [variously paged].

U.S. Environmental Protection Agency (EPA), 2009b, Method 1600 “Enterococci in water by membrane filtrations using membrane-Enterococcus indoxyl-β-D-glucoside agar”: Washington, D.C., EPA–821–R–09–016 [variously paged].

U.S. Environmental Protection Agency (EPA), 2012, Recre-ational water quality criteria: Office of Water, EPA 815–F–12–058, 63 p.

U.S. Environmental Protection Agency (EPA) and U.S. Geological Survey (USGS), 2012, National Hydrography Dataset Plus - NHDPlus version 2.1, accessed May 2012 at http://www.horizon-systems.com/NHDPlus/NHDPlusV2_home.php.

U.S. Geological Survey (USGS), 2006, Collection of water samples (ver. 2.0): U.S. Geological Survey Techniques of Water-Resources Investigations, book 9, chap. A4, Septem-ber 2006, accessed June 1, 2014, at http://pubs.water.usgs.gov/twri9A4/.

U.S. Geological Survey (USGS), 2011, National Land Cover Database (NLCD) 2006 Land Cover, accessed April 3, 2012, at http://www.mrlc.gov/nlcd06_data.php.

U.S. Geological Survey (USGS), Great Lakes Restoration Ini-tiative (GLRI), 2014, Forecast/nowcast Great Lakes nutrient and sediment loadings: USGS/GLRI Web page, accessed March 6, 2014, at http://cida.usgs.gov/glri/projects/near-shore_health/forecast_loadings.html.

http://www.cdc.gov/nczved/dfbmd/diseases/campylobacter/

http://www.cdc.gov/salmonella

http://www.cdc.gov/ecoli/index.html

http://www.cdc.gov/ecoli/index.html

http://www.glin.net/lakes/ref/lakefact.html

http://www.horizon-systems.com/NHDPlus/NHDPlusV2_home.php

http://www.horizon-systems.com/NHDPlus/NHDPlusV2_home.php

http://www.mrlc.gov/nlcd06_data.php

http://cida.usgs.gov/glri/projects/nearshore_health/forecast_loadings.html

http://cida.usgs.gov/glri/projects/nearshore_health/forecast_loadings.html

Appendix 13

Appendixes

1. Fecal indicator bacteria concentrations in Great Lakes tributaries, 2011 ........................14 2. Quality-control data—Fecal indicator bacteria concentrations in Great Lakes

tributaries, 2011 ...........................................................................................................................20 3. Pathogen gene detection in Great Lakes tributaries, 2011 ..................................................22 4. Quality-control data—Pathogen gene detection in Great Lakes tributaries, 2011 .........28

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ular

150

E36

E42

0415

7000

SG-0

67/

14/2

011

Nor

mal

Reg

ular

950

E25

E12

E04

1570

00SG

-07

7/14

/201

1N

orm

alR

eplic

ate

SG-0

685

0E

20E

13E

0415

7000

SG-0

88/

23/2

011

Nor

mal

Reg

ular

5230

E12

E

0415

7000

SG-1

19/

14/2

011

Nor

mal

Reg

ular

110

6040

0410

1500

SJ-0

24/

7/20

11N

orm

alR

egul

ar26

E2

<7

E

0410

1500

SJ-0

77/

27/2

011

Nor

mal

Reg

ular

110

7452

0410

1500

SJ-0

88/

24/2

011

Nor

mal

Reg

ular

370

E13

052

0426

9000

SR-0

13/

24/2

011

Nor

mal

Reg

ular

16E

6E

2E

0426

9000

SR-0

44/

26/2

011

Nor

mal

Reg

ular

14E

8E

2E

0426

9000

SR-0

76/

21/2

011

Nor

mal

Reg

ular

nsns

ns

0426

9000

SR-0

87/

26/2

011

Nor

mal

Reg

ular

170

E13

046

0426

9000

SR-0

97/

26/2

011

Nor

mal

Rep

licat

eSR

-08

210

E14

052

0426

9000

SR-1

39/

20/2

011

Nor

mal

Reg

ular

150

E18

0E

110

0402

4000

ST-0

13/

15/2

011

Nor

mal

Reg

ular

8E

2E

2E

0402

4000

ST-0

24/

20/2

011

Nor

mal

Reg

ular

2E

4E

5E

0402

4000

ST-0

46/

14/2

011

Nor

mal

Reg

ular

12E

3E

2E

0402

4000

ST-0

56/

23/2

011

Nor

mal

Reg

ular

26E

18E

44E

0402

4000

ST-0

67/

21/2

011

Nor

mal

Reg

ular

120

26E

28E

0402

4000

ST-0

77/

21/2

011

Nor

mal

Rep

licat

eST

-06

110

4242

0402

4000

ST-1

18/

31/2

011

Nor

mal

Reg

ular

7440

20E

0402

4000

ST-1

29/

13/2

011

Nor

mal

Reg

ular

4225

E10

E

0413

7500

AU

-02

4/12

/201

1H

igh

Reg

ular

2<

2<

2<

0402

7000

BD

-01

4/11

/201

1H

igh

Reg

ular

110

82E

280

0402

7000

BD

-02

4/11

/201

1H

igh

Rep

licat

eB

D-0

114

0E

5627

0

Appendix 17A

ppen

dix

1.

Feca

l ind

icat

or b

acte

ria c

once

ntra

tions

in G

reat

Lak

es tr

ibut

arie

s, 2

011.

—Co

ntin

ued

[Stre

amga

ging

stat

ions

cor

resp

ondi

ng to

stat

ion

IDs a

nd fi

eld

IDs a

re li

sted

in ta

ble

1 in

the

mai

n re

port.

USG

S, U

.S. G

eolo

gica

l Sur

vey;

Ass

oc. e

nviro

n. sa

mpl

e, a

ssoc

iate

d en

viro

nmen

-ta

l sam

ple;

E. c

oli,

Esch

eric

hia

coli;

CFU

/100

mL,

col

ony-

form

ing

units

per

100

mill

ilite

rs; n

s, no

sam

ple;

unc

, unc

ount

able

; E, e

stim

ated

val

ue; <

, les

s tha

n; >

, gre

ater

than

]

Sam

ple

A

ssoc

.Fe

cal

E. c

oli

USG

S

Fiel

d

Flow

Sa

mpl

e

Rem

ark

Rem

ark

Ente

roco

cci

Rem

ark

colle

ctio

n

envi

ron.

colif

orm

s(C

FU/

stat

ion

IDID

cond

ition

sty

peco

deco

de(C

FU/1

00 m

L)co

deda

tesa

mpl

e(C

FU/1

00m

L)10

0 m

L)

0402

7000

BD

-03

5/24

/201

1H

igh

Reg

ular

340

E51

015

0E

0409

5090

BN

-02

4/28

/201

1H

igh

Reg

ular

120

>14

00E

1100

E

0409

5090

BN

-03

5/25

/201

1H

igh

Reg

ular

610

E71

0E

350

0409

5090

BN

-04

6/8/

2011

Hig

hR

egul

ar46

17E

7E

0409

5090

BN

-10

9/7/

2011

Hig

hR

egul

ar94

4856

0416

5500

CL-

015/

16/2

011

Hig

hR

egul

ar12

0>

2500

3800

0416

5500

CL-

028/

17/2

011

Hig

hR

egul

ar23

0E

200

100

0416

5500

CL-

038/

17/2

011

Hig

hR

eplic

ate

CL-

0216

0E

140

84

0420

8000

CY-

014/

27/2

011

Hig

hR

egul

ar49

0E

590

250

0420

8000

CY-

027/

26/2

011

Hig

hR

egul

ar12

0>

180

100

0420

8000

CY-

037/

26/2

011

Hig

hR

eplic

ate

CY-

0212

0>

250

78

0420

8000

CY-

079/

20/2

011

Hig

hR

egul

ar12

0>

6100

8000

E

0420

8000

CY-

0810

/26/

2011

Hig

hR

egul

ar43

0E

430

420

0408

5138

5FX

-01

5/4/

2011

Hig

hR

egul

ar14

E64

4E

0411

9400

GR

-01

4/29

/201

1H

igh

Reg

ular

310

E29

028

0

0411

9400

GR

-02

5/18

/201

1H

igh

Reg

ular

140

E23

034

E

0411

9400

GR

-03

6/2/

2011

Hig

hR

egul

ar70

4817

E

0411

9400

GR

-04

6/29

/201

1H

igh

Reg

ular

240

E82

12E

0411

9400

GR

-05

6/29

/201

1H

igh

Rep

licat

eG

R-0

427

0E

808

E

0411

9400

GR

-06

8/2/

2011

Hig

hR

egul

ar13

0E

9017

E

0423

1600

GS-

024/

26/2

011

Hig

hR

egul

ar40

0E

320

270

0423

1600

GS-

035/

17/2

011

Hig

hR

egul

ar12

0>

1200

E14

00E

0423

1600

GS-

046/

2/20

11H

igh

Reg

ular

unc

390

120

0419

3500

MA

-01

3/8/

2011

Hig

hR

egul

ar57

0E

650

980

E

0408

5427

MN

-02

7/6/

2011

Hig

hR

egul

ar41

0E

7050

0408

7170

MW

-01

5/3/

2011

Hig

hR

egul

ar28

E58

27E


ppen

dix

1.

Feca

l ind

icat

or b

acte

ria c

once

ntra

tions

in G

reat

Lak

es tr

ibut

arie

s, 2

011.

—Co

ntin

ued

[Stre

amga

ging

stat

ions

cor

resp

ondi

ng to

stat

ion

IDs a

nd fi

eld

IDs a

re li

sted

in ta

ble

1 in

the

mai

n re

port.

USG

S, U

.S. G

eolo

gica

l Sur

vey;

Ass

oc. e

nviro

n. sa

mpl

e, a

ssoc

iate

d en

viro

nmen

-ta

l sam

ple;

E. c

oli,

Esch

eric

hia

coli;

CFU

/100

mL,

col

ony-

form

ing

units

per

100

mill

ilite

rs; n

s, no

sam

ple;

unc

, unc

ount

able

; E, e

stim

ated

val

ue; <

, les

s tha

n; >

, gre

ater

than

]

Sam

ple

A

ssoc

.Fe

cal

E. c

oli

USG

S

Fiel

d

Flow

Sa

mpl

e

Rem

ark

Rem

ark

Ente

roco

cci

Rem

ark

colle

ctio

n

envi

ron.

colif

orm

s(C

FU/

stat

ion

IDID

cond

ition

sty

peco

deco

de(C

FU/1

00 m

L)co

deda

tesa

mpl

e(C

FU/1

00m

L)10

0 m

L)

0404

0000

ON

-06

6/23

/201

1H

igh

Reg

ular

120

>92

0E

570

0424

9000

OS-

013/

9/20

11H

igh

Reg

ular

150

E10

064

0424

9000

OS-

034/

27/2

011

Hig

hR

egul

ar70

8866

0424

9000

OS-

045/

9/20

11H

igh

Reg

ular

4E

18E

2<

0424

9000

OS-

159/

7/20

11H

igh

Reg

ular

7015

E11

E

0419

5500

PO-0

13/

8/20

11H

igh

Reg

ular

580

E65

057

0

0419

5500

PO-0

24/

20/2

011

Hig

hR

egul

ar12

0>

1400

E36

0004

1955

00PO

-03

7/27

/201

1H

igh

Reg

ular

120

>24

031

0

0419

5500

PO-0

47/

27/2

011

Hig

hR

eplic

ate

PO-0

312

0>

220

280

0410

2500

PP-0

24/

21/2

011

Hig

hR

egul

ar12

0>

790

1200

E

0410

2500

PP-0

36/

1/20

11H

igh

Reg

ular

9610

072

0410

2500

PP-0

47/

19/2

011

Hig

hR

egul

ar12

0>

4800

6700

E

0410

2500

PP-0

58/

3/20

11H

igh

Reg

ular

410

E63

042

0E

0410

2500

PP-0

810

/4/2

011

Hig

hR

egul

ar12

070

82

0416

6500

RR

-01

5/16

/201

1H

igh

Reg

ular

120

>26

0057

00

0416

6500

RR

-02

5/26

/201

1H

igh

Reg

ular

120

>26

000

E31

000

E

0415

7000

SG-0

24/

6/20

11H

igh

Reg

ular

240

E22

044

0

0415

7000

SG-0

35/

2/20

11H

igh

Reg

ular

100

190

E98

0415

7000

SG-0

45/

16/2

011

Hig

hR

egul

ar12

0>

1200

E96

0E

0410

1500

SJ-0

13/

9/20

11H

igh

Reg

ular

unc

9644

0

0410

1500

SJ-0

34/

21/2

011

Hig

hR

egul

ar12

0>

860

E13

00E

0410

1500

SJ-0

44/

28/2

011

Hig

hR

egul

ar12

0>

1400

E87

0E

0410

1500

SJ-0

56/

28/2

011

Hig

hR

eplic

ate

SJ-0

626

0E

100

23

0410

1500

SJ-0

66/

28/2

011

Hig

hR

egul

ar38

0E

5840

0426

9000

SR-0

24/

6/20

11H

igh

Reg

ular

16E

5E

14E

0426

9000

SR-0

34/

12/2

011

Hig

hR

egul

ar50

40E

33E

Appendix 19A

ppen

dix

1.

Feca

l ind

icat

or b

acte

ria c

once

ntra

tions

in G

reat

Lak

es tr

ibut

arie

s, 2

011.

—Co

ntin

ued

[Stre

amga

ging

stat

ions

cor

resp

ondi

ng to

stat

ion

IDs a

nd fi

eld

IDs a

re li

sted

in ta

ble

1 in

the

mai

n re

port.

USG

S, U

.S. G

eolo

gica

l Sur

vey;

Ass

oc. e

nviro

n. sa

mpl

e, a

ssoc

iate

d en

viro

nmen

-ta

l sam

ple;

E. c

oli,

Esch

eric

hia

coli;

CFU

/100

mL,

col

ony-

form

ing

units

per

100

mill

ilite

rs; n

s, no

sam

ple;

unc

, unc

ount

able

; E, e

stim

ated

val

ue; <

, les

s tha

n; >

, gre

ater

than

]

Sam

ple

A

ssoc

.Fe

cal

E. c

oli

USG

S

Fiel

d

Flow

Sa

mpl

e

Rem

ark

Rem

ark

Ente

roco

cci

Rem

ark

colle

ctio

n

envi

ron.

colif

orm

s(C

FU/

stat

ion

IDID

cond

ition

sty

peco

deco

de(C

FU/1

00 m

L)co

deda

tesa

mpl

e(C

FU/1

00m

L)10

0 m

L)

0426

9000

SR-0

55/

5/20

11H

igh

Reg

ular

24E

4617

E

0426

9000

SR-0

65/

24/2

011

Hig

hR

egul

ar30

E60

30E

0426

9000

SR-1

28/

23/2

011

Hig

hR

egul

ar22

0E

220

210

0402

4000

ST-0

35/

1/20

11H

igh

Reg

ular

16E

17E

6E

0402

4000

ST-1

08/

3/20

11H

igh

Reg

ular

580

E79

036

0


ppen

dix

2.

Qual

ity-c

ontro

l dat

a—Fe

cal i

ndic

ator

bac

teria

con

cent

ratio

ns in

Gre

at L

akes

trib

utar

ies,

201

1.—

Cont

inue

d

[Stre

amga

ging

stat

ions

cor

resp

ondi

ng to

stat

ion

IDs a

nd fi

eld

IDs a

re li

sted

in ta

ble

1 in

the

mai

n re

port.

USG

S, U

.S. G

eolo

gica

l Sur

vey;

Ass

oc. e

nviro

n. sa

mpl

e, a

ssoc

iate

d en

viro

nmen

tal s

ampl

e; E

. col

i, Es

cher

ichi

a co

li; C

FU/1

00 m

L, c

olon

y-fo

rmin

g un

its p

er 1

00 m

illili

ters

; E, e

stim

ated

val

ue; <

, les

s tha

n; >

, gre

ater

than

; N/A

; not

app

licab

le; n

s, no

sam

ple]

USG

S

stat

ion

IDFi

eld

ID

Sam

ple

co

llect

ion

da

teSa

mpl

e ty

peA

ssoc

. en

viro

n.

sam

ple

Feca

l co

lifor

ms

(CFU

/100

mL)

Rem

ark

code

E. c

oli

(CFU

/100

mL)

Rem

ark

code

Ente

ro-c

occi

(C

FU/1

00 m

L)Re

mar

k co

de

0413

7500

AU

-08

8/23

/201

1Fi

eld

blan

kA

U-0

72

<2

<2

<04

0270

00B

D-0

67/

12/2

011

Fiel

d bl

ank

BD

-04

2<

2<

2<

0409

5090

BN

-06

7/12

/201

1Fi

eld

blan

kB

N-0

52

<2

<2

<04

1655

00C

L-04

11/1

/201

1Fi

eld

blan

kns

2<

2<

2<

0420

8000

CY-

047/

26/2

011

Fiel

d bl

ank

CY-

022

<2

<2

<04

0851

385

FX-0

49/

19/2

011

Fiel

d bl

ank

ns2

<2

<2

<04

1194

00G

R-0

710

/31/

2011

Fiel

d bl

ank

ns2

<2

<2

<04

2316

00G

S-11

7/18

/201

1Fi

eld

blan

kG

S-09

2<

2<

2<

0419

3500

MA

-05

7/27

/201

1Fi

eld

blan

kM

A-0

32

<2

<2

<04

0675

00M

M-0

59/

7/20

11Fi

eld

blan

kM

M-0

32

<2

<2

<04

0854

27M

N-0

59/

8/20

11Fi

eld

blan

kM

N-0

32

<2

<2

<04

0871

70M

W-0

49/

20/2

011

Fiel

d bl

ank

MW

-02

2<

2<

2<

0404

0000

ON

-12

9/20

/201

1Fi

eld

blan

kns

2<

2<

2<

0424

9000

OS-

127/

18/2

011

Fiel

d bl

ank

OS-

102

<2

<2

<04

1955

00PO

-05

7/27

/201

1Fi

eld

blan

kPO

-03

2<

2<

2<

0416

6500

RR

-05

10/3

1/20

11Fi

eld

blan

kns

2<

2<

2<

0417

6500

RS-

048/

30/2

011

Fiel

d bl

ank

ns2

<2

<2

E04

1570

00SG

-09

8/23

/201

1Fi

eld

blan

kSG

-08

2<

2<

2<

0410

1500

SJ-0

98/

24/2

011

Fiel

d bl

ank

SJ-0

82

<2

<2

<04

2690

00SR

-10

7/26

/201

1Fi

eld

blan

kSR

-08

2<

2<

2<

0402

4000

ST-0

87/

21/2

011

Fiel

d bl

ank

ST-0

62

<2

<2

<04

1375

00A

U-0

98/

22/2

011

Sour

ce so

lutio

n bl

ank

ns2

<2

<2

<04

0270

00B

D-0

77/

12/2

011

Sour

ce so

lutio

n bl

ank

BD

-04

2<

2<

2<

0409

5090

BN

-07

7/12

/201

1So

urce

solu

tion

blan

kB

N-0

52

<2

<2

<04

1655

00C

L-05

11/1

/201

1So

urce

solu

tion

blan

kns

2<

2<

2<

0420

8000

CY-

057/

26/2

011

Sour

ce so

lutio

n bl

ank

CY-

022

<2

<2

<

Appendix 21A

ppen

dix

2.

Qual

ity-c

ontro

l dat

a—Fe

cal i

ndic

ator

bac

teria

con

cent

ratio

ns in

Gre

at L

akes

trib

utar

ies,

201

1.—

Cont

inue

d

[Stre

amga

ging

stat

ions

cor

resp

ondi

ng to

stat

ion

IDs a

nd fi

eld

IDs a

re li

sted

in ta

ble

1 in

the

mai

n re

port.

USG

S, U

.S. G

eolo

gica

l Sur

vey;

Ass

oc. e

nviro

n. sa

mpl

e, a

ssoc

iate

d en

viro

nmen

tal s

ampl

e; E

. col

i, Es

cher

ichi

a co

li; C

FU/1

00 m

L, c

olon

y-fo

rmin

g un

its p

er 1

00 m

illili

ters

; E, e

stim

ated

val

ue; <

, les

s tha

n; >

, gre

ater

than

; N/A

; not

app

licab

le; n

s, no

sam

ple]

Sam

ple

A

ssoc

. Fe

cal

USG

S

Fiel

d

Rem

ark

E. c

oli

Rem

ark

Ente

ro-c

occi

Re

mar

k co

llect

ion

Sa

mpl

e ty

peen

viro

n.

colif

orm

s st

atio

n ID

IDco

de(C

FU/1

00 m

L)co

de(C

FU/1

00 m

L)co

deda

tesa

mpl

e(C

FU/1

00 m

L)

0408

5138

5FX

-05

9/19

/201

1So

urce

solu

tion

blan

kns

2<

2<

2<

0411

9400

GR

-08

10/3

1/20

11So

urce

solu

tion

blan

kns

2<

2<

2<

0423

1600

GS-

127/

18/2

011

Sour

ce so

lutio

n bl

ank

GS-

092

<2

<2

<04

1935

00M

A-0

67/

27/2

011

Sour

ce so

lutio

n bl

ank

MA

-03

2<

2<

2<

0406

7500

MM

-06

9/19

/201

1So

urce

solu

tion

blan

kns

2<

2<

2<

0408

5427

MN

-06

9/19

/201

1So

urce

solu

tion

blan

kns

2<

2<

2<

0408

7170

MW

-05

9/20

/201

1So

urce

solu

tion

blan

kM

W-0

22

<2

<2

<04

0400

00O

N-1

19/

20/2

011

Sour

ce so

lutio

n bl

ank

ns2

<2

<2

<04

2490

00O

S-13

7/18

/201

1So

urce

solu

tion

blan

kO

S-10

2<

2<

2<

0419

5500

PO-0

67/

27/2

011

Sour

ce so

lutio

n bl

ank

PO-0

32

<2

<2

<04

1665

00R

R-0

610

/31/

2011

Sour

ce so

lutio

n bl

ank

ns2

<2

<2

<04

1765

00R

S-05

8/30

/201

1So

urce

solu

tion

blan

kns

2<

2<

2<

0415

7000

SG-1

08/

23/2

011

Sour

ce so

lutio

n bl

ank

SG-0

82

<2

<2

<04

1015

00SJ

-10

8/24

/201

1So

urce

solu

tion

blan

kSJ

-08

2<

2<

2<

0426

9000

SR-1

17/

26/2

011

Sour

ce so

lutio

n bl

ank

SR-0

82

<2

<2

<04

0240

00ST

-09

7/21

/201

1So

urce

solu

tion

blan

kST

-06

2<

2<

2<


ppen

dix

3.

Path

ogen

gen

e de

tect

ion

in G

reat

Lak

es tr

ibut

arie

s, 2

011.

—Co

ntin

ued

[Stre

amga

ging

stat

ions

cor

resp

ondi

ng to

stat

ion

IDs a

nd fi

eld

IDs a

re li

sted

in ta

ble

1 in

the

mai

n re

port.

USG

S, U

.S. G

eolo

gica

l Sur

vey;

Ass

oc. e

nviro

n. sa

mpl

e, a

ssoc

iate

d en

viro

nmen

tal s

ampl

e; E

. col

i, Es

cher

ichi

a co

li; +

, gen

e pr

esen

t in

sam

ple;

—, g

ene

abse

nt in

sam

ple;

ns,

no sa

mpl

e; V

– re

mar

k co

de in

dica

ting

poss

ible

con

tam

inat

ion

(ana

lyte

was

det

ecte

d in

bot

h th

e en

viro

nmen

tal s

ampl

e an

d th

e as

soci

ated

bla

nks)

]

USG

S

stat

ion

IDFi

eld

ID

Sam

ple

co

llect

ion

da

te

Flow

co

nditi

ons

Sam

ple

type

Ass

oc.

e

nvir

on.

sam

ple

E. c

oli g

ene

data

Shig

ella

ge

ne d

ata

Salm

onel

la

gene

dat

aCa

mpy

loba

cter

ge

ne d

ata

eaeA

stx2

stx1

rfb

O

157

ipaH

invA

spvC

C. c

oli/

C. je

juni

0413

7500

AU

-01

3/15

/201

1N

orm

alR

egul

ar —

——

——

——

—04

1375

00A

U-0

46/

15/2

011

Nor

mal

Reg

ular

——

——

——

——

0413

7500

AU

-05

7/13

/201

1N

orm

alR

egul

ar+

——

——

——

—04

1375

00A

U-0

67/

13/2

011

Nor

mal

Rep

licat

eA

U-0

5+

——

——

——

—

0413

7500

AU

-07

8/23

/201

1N

orm

alR

egul

ar—

——

—+

——

—04

1375

00A

U-1

09/

13/2

011

Nor

mal

Reg

ular

——

——

——

——

0402

7000

BD

-04

7/12

/201

1N

orm

alR

egul

ar+

——

——

——

—04

0270

00B

D-0

57/

12/2

011

Nor

mal

Rep

licat

eB

-04

+—

——

——

——

0402

7000

BD

-08

8/16

/201

1N

orm

alR

egul

ar+

——

——

——

—04

0270

00B

D-0

99/

20/2

011

Nor

mal

Reg

ular

+—

——

——

+—

0409

5090

BN

-01

3/30

/201

1N

orm

alR

egul

ar+

——

——

——

—04

0950

90B

N-0

57/

12/2

011

Nor

mal

Reg

ular

+—

——

——

+—

0409

5090

BN

-08

8/11

/201

1N

orm

alR

egul

ar+

——

——

——

—04

0950

90B

N-0

98/

11/2

011

Nor

mal

Rep

licat

eB

N-0

8+

——

——

——

—

0420

8000

CY-

068/

31/2

011

Nor

mal

Reg

ular

+—

——

——

—+

0408

5138

5FX

-02

8/16

/201

1N

orm

alR

egul

ar—

——

——

——

—04

0851

385

FX-0

38/

16/2

011

Nor

mal

Rep

licat

eFX

-02

——

——

——

——

0423

1600

GS-

014/

5/20

11N

orm

alR

egul

ar+

——

——

——

+04

2316

00G

S-09

7/18

/201

1N

orm

alR

egul

ar—

——

——

——

—04

2316

00G

S-10

7/18

/201

1N

orm

alR

eplic

ate

GS-

09—

——

——

+—

—

0423

1600

GS-

138/

8/20

11N

orm

alR

egul

ar—

——

——

——

—04

1935

00M

A-0

24/

19/2

011

Nor

mal

Reg

ular

+—

——

——

——

0419

3500

MA

-03

7/27

/201

1N

orm

alR

egul

ar+

——

+—

——

—04

1935

00M

A-0

47/

27/2

011

Nor

mal

Rep

licat

eM

A-0

3+

——

+—

——

—

0406

7500

MM

-02

7/7/

2011

Nor

mal

Reg

ular

——

——

——

——

Appendix 23A

ppen

dix

3.

Path

ogen

gen

e de

tect

ion

in G

reat

Lak

es tr

ibut

arie

s, 2

011.

—Co

ntin

ued

[Stre

amga

ging

stat

ions

cor

resp

ondi

ng to

stat

ion

IDs a

nd fi

eld

IDs a

re li

sted

in ta

ble

1 in

the

mai

n re

port.

USG

S, U

.S. G

eolo

gica

l Sur

vey;

Ass

oc. e

nviro

n. sa

mpl

e, a

ssoc

iate

d en

viro

nmen

tal s

ampl

e; E

. col

i, Es

cher

ichi

a co

li; +

, gen

e pr

esen

t in

sam

ple;

—, g

ene

abse

nt in

sam

ple;

ns,

no sa

mpl

e; V

– re

mar

k co

de in

dica

ting

poss

ible

con

tam

inat

ion

(ana

lyte

was

det

ecte

d in

bot

h th

e en

viro

nmen

tal s

ampl

e an

d th

e as

soci

ated

bla

nks)

]

Shig

ella

Sa

lmon

ella

Ca

mpy

loba

cter

E.

col

i gen

e da

taSa

mpl

e

Ass

oc.

ge

ne d

ata

gene

dat

age

ne d

ata

USG

S

Fiel

d

Flow

co

llect

ion

Sa

mpl

e ty

pe e

nvir

on.

stat

ion

IDID

cond

ition

sda

te s

ampl

erf

b

C. c

oli/

eaeA

stx2

stx1

ipaH

invA

spvC

O15

7C.

jeju

ni

0406

7500

MM

-03

9/7/

2011

Nor

mal

Reg

ular

+V—

——

——

——

0406

7500

MM

-04

9/7/

2011

Nor

mal

Rep

licat

eM

M-0

3+

——

+—

——

—

0408

5427

MN

-03

9/8/

2011

Nor

mal

Reg

ular

——

——

——

—+

0408

5427

MN

-04

9/8/

2011

Nor

mal

Rep

licat

eM

N-0

3+

——

——

——

+

0408

7170

MW

-02

9/20

/201

1N

orm

alR

egul

ar+

——

+—

——

—04

0871

70M

W-0

39/

20/2

011

Nor

mal

Rep

licat

eM

W-0

2+

——

——

+—

—

0404

0000

ON

-01

3/23

/201

1N

orm

alR

egul

ar—

—+

——

——

—04

0400

00O

N-0

24/

5/20

11N

orm

alR

egul

ar+

——

——

——

—04

0400

00O

N-0

34/

26/2

011

Nor

mal

Reg

ular

——

——

——

——

0404

0000

ON

-04

5/25

/201

1N

orm

alR

egul

ar+

——

——

——

—04

0400

00O

N-0

56/

6/20

11N

orm

alR

egul

ar+

——

——

——

—04

0400

00O

N-0

77/

14/2

011

Nor

mal

Reg

ular

——

——

——

——

0404

0000

ON

-08

8/17

/201

1N

orm

alR

egul

ar—

——

—+

——

—04

0400

00O

N-0

99/

13/2

011

Nor

mal

Reg

ular

+—

——

——

——

0404

0000

ON

-10

9/13

/201

1N

orm

alR

eplic

ate

ON

-09

+—

——

——

——

0424

9000

OS-

024/

7/20

11N

orm

alR

egul

ar—

——

——

——

—04

2490

00O

S-05

6/8/

2011

Nor

mal

Reg

ular

+—

——

——

——

0424

9000

OS-

107/

18/2

011

Nor

mal

Reg

ular

——

——

——

——

0424

9000

OS-

117/

18/2

011

Nor

mal

Rep

licat

eO

S-10

+—

——

——

——

0424

9000

OS-

148/

8/20

11N

orm

alR

egul

ar+

——

——

——

—04

1025

00PP

-01

4/11

/201

1N

orm

alR

egul

ar+

+—

——

——

—04

1025

00PP

-06

9/8/

2011

Nor

mal

Reg

ular

++

——

——

++

0416

6500

RR

-03

8/17

/201

1N

orm

alR

egul

ar+

——

+—

—+

—04

1665

00R

R-0

48/

17/2

011

Nor

mal

Rep

licat

eR

R-0

3+

——

——

—+

+

0417

6500

RS-

016/

16/2

011

Nor

mal

Reg

ular

++

——

——

——


ppen

dix

3.

Path

ogen

gen

e de

tect

ion

in G

reat

Lak

es tr

ibut

arie

s, 2

011.

—Co

ntin

ued

[Stre

amga

ging

stat

ions

cor

resp

ondi

ng to

stat

ion

IDs a

nd fi

eld

IDs a

re li

sted

in ta

ble

1 in

the

mai

n re

port.

USG

S, U

.S. G

eolo

gica

l Sur

vey;

Ass

oc. e

nviro

n. sa

mpl

e, a

ssoc

iate

d en

viro

nmen

tal s

ampl

e; E

. col

i, Es

cher

ichi

a co

li; +

, gen

e pr

esen

t in

sam

ple;

—, g

ene

abse

nt in

sam

ple;

ns,

no sa

mpl

e; V

– re

mar

k co

de in

dica

ting

poss

ible

con

tam

inat

ion

(ana

lyte

was

det

ecte

d in

bot

h th

e en

viro

nmen

tal s

ampl

e an

d th

e as

soci

ated

bla

nks)

]

Shig

ella

Sa

lmon

ella

Ca

mpy

loba

cter

E.

col

i gen

e da

taSa

mpl

e

Ass

oc.

ge

ne d

ata

gene

dat

age

ne d

ata

USG

S

Fiel

d

Flow

co

llect

ion

Sa

mpl

e ty

pe e

nvir

on.

stat

ion

IDID

cond

ition

sda

te s

ampl

erf

b

C. c

oli/

eaeA

stx2

stx1

ipaH

invA

spvC

O15

7C.

jeju

ni

0417

6500

RS-

028/

23/2

011

Nor

mal

Reg

ular

+—

——

+—

——

0417

6500

RS-

038/

23/2

011

Nor

mal

Rep

licat

eR

S-02

+—

——

+—

——

0415

7000

SG-0

13/

15/2

011

Nor

mal

Reg

ular

+—

——

——

——

0415

7000

SG-0

56/

16/2

011

Nor

mal

Reg

ular

——

——

——

——

0415

7000

SG-0

67/

14/2

011

Nor

mal

Reg

ular

——

——

——

——

0415

7000

SG-0

77/

14/2

011

Nor

mal

Rep

licat

eSG

-06

+—

——

——

——

0415

7000

SG-0

88/

23/2

011

Nor

mal

Reg

ular

+—

——

——

——

0415

7000

SG-1

19/

14/2

011

Nor

mal

Reg

ular

+—

——

——

——

0410

1500

SJ-0

24/

7/20

11N

orm

alR

egul

ar—

——

——

——

—04

1015

00SJ

-07

7/27

/201

1N

orm

alR

egul

ar+

——

——

——

—04

1015

00SJ

-08

8/24

/201

1N

orm

alR

egul

ar+V

——

——

——

—04

2690

00SR

-01

3/24

/201

1N

orm

alR

egul

ar—

—+

——

——

—04

2690

00SR

-04

4/26

/201

1N

orm

alR

egul

ar—

——

——

——

—04

2690

00SR

-07

6/21

/201

1N

orm

alR

egul

ar+

——

——

——

—04

2690

00SR

-08

7/26

/201

1N

orm

alR

egul

ar+

——

——

——

—04

2690

00SR

-09

7/26

/201

1N

orm

alR

eplic

ate

SR-0

8+

——

——

——

—

0426

9000

SR-1

39/

20/2

011

Nor

mal

Reg

ular

+—

——

——

+—

0402

4000

ST-0

13/

15/2

011

Nor

mal

Reg

ular

——

——

——

——

0402

4000

ST-0

24/

20/2

011

Nor

mal

Reg

ular

——

——

——

——

0402

4000

ST-0

46/

14/2

011

Nor

mal

Reg

ular

++

——

——

——

0402

4000

ST-0

56/

23/2

011

Nor

mal

Reg

ular

+—

——

——

——

0402

4000

ST-0

67/

21/2

011

Nor

mal

Reg

ular

——

——

——

——

0402

4000

ST-0

77/

21/2

011

Nor

mal

Rep

licat

eST

-06

+—

——

——

——

0402

4000

ST-1

18/

31/2

011

Nor

mal

Reg

ular

——

——

——

—+

0402

4000

ST-1

29/

13/2

011

Nor

mal

Reg

ular

+—

——

——

——

Appendix 25A

ppen

dix

3.

Path

ogen

gen

e de

tect

ion

in G

reat

Lak

es tr

ibut

arie

s, 2

011.

—Co

ntin

ued

[Stre

amga

ging

stat

ions

cor

resp

ondi

ng to

stat

ion

IDs a

nd fi

eld

IDs a

re li

sted

in ta

ble

1 in

the

mai

n re

port.

USG

S, U

.S. G

eolo

gica

l Sur

vey;

Ass

oc. e

nviro

n. sa

mpl

e, a

ssoc

iate

d en

viro

nmen

tal s

ampl

e; E

. col

i, Es

cher

ichi

a co

li; +

, gen

e pr

esen

t in

sam

ple;

—, g

ene

abse

nt in

sam

ple;

ns,

no sa

mpl

e; V

– re

mar

k co

de in

dica

ting

poss

ible

con

tam

inat

ion

(ana

lyte

was

det

ecte

d in

bot

h th

e en

viro

nmen

tal s

ampl

e an

d th

e as

soci

ated

bla

nks)

]

Shig

ella

Sa

lmon

ella

Ca

mpy

loba

cter

E.

col

i gen

e da

taSa

mpl

e

Ass

oc.

ge

ne d

ata

gene

dat

age

ne d

ata

USG

S

Fiel

d

Flow

co

llect

ion

Sa

mpl

e ty

pe e

nvir

on.

stat

ion

IDID

cond

ition

sda

te s

ampl

erf

b

C. c

oli/

eaeA

stx2

stx1

ipaH

invA

spvC

O15

7C.

jeju

ni

0413

7500

AU

-02

4/12

/201

1H

igh

Reg

ular

——

——

——

——

0402

7000

BD

-01

4/11

/201

1H

igh

Reg

ular

——

——

——

——

0402

7000

BD

-02

4/11

/201

1H

igh

Rep

licat

eB

D-0

1—

——

——

——

—

0402

7000

BD

-03

5/24

/201

1H

igh

Reg

ular

+—

——

——

——

0409

5090

BN

-02

4/28

/201

1H

igh

Reg

ular

+—

++

—+

—+

0409

5090

BN

-03

5/25

/201

1H

igh

Reg

ular

+—

——

—+

——

0409

5090

BN

-04

6/8/

2011

Hig

hR

egul

ar—

——

——

——

—04

0950

90B

N-1

09/

7/20

11H

igh

Reg

ular

+—

——

——

——

0416

5500

CL-

015/

16/2

011

Hig

hR

egul

ar+

++

+—

—+

—04

1655

00C

L-02

8/17

/201

1H

igh

Reg

ular

+—

——

——

+—

0416

5500

CL-

038/

17/2

011

Hig

hR

eplic

ate

CL-

02+

——

+—

—+

—

0420

8000

CY-

014/

27/2

011

Hig

hR

egul

ar—

——

——

——

—04

2080

00C

Y-02

7/26

/201

1H

igh

Reg

ular

+—

——

——

——

0420

8000

CY-

037/

26/2

011

Hig

hR

eplic

ate

CY-

02+

+—

——

——

—

0420

8000

CY-

079/

20/2

011

Hig

hR

egul

ar+

++

+—

—+

—04

2080

00C

Y-08

10/2

6/20

11H

igh

Reg

ular

+—

——

——

——

0408

5138

5FX

-01

5/4/

2011

Hig

hR

egul

ar+

——

——

——

—04

1194

00G

R-0

14/

29/2

011

Hig

hR

egul

ar+

+—

——

——

—04

1194

00G

R-0

25/

18/2

011

Hig

hR

egul

ar+

——

——

——

—04

1194

00G

R-0

36/

2/20

11H

igh

Reg

ular

+—

——

+—

——

0411

9400

GR

-04

6/29

/201

1H

igh

Reg

ular

+—

——

——

+—

0411

9400

GR

-05

6/29

/201

1H

igh

Rep

licat

eG

R-0

4+

—+

——

——

—

0411

9400

GR

-06

8/2/

2011

Hig

hR

egul

ar+

++

—+

——

—04

2316

00G

S-02

4/26

/201

1H

igh

Reg

ular

+—

+—

——

——

0423

1600

GS-

035/

17/2

011

Hig

hR

egul

ar+

—+

+—

——

—


ppen

dix

3.

Path

ogen

gen

e de

tect

ion

in G

reat

Lak

es tr

ibut

arie

s, 2

011.

—Co

ntin

ued

[Stre

amga

ging

stat

ions

cor

resp

ondi

ng to

stat

ion

IDs a

nd fi

eld

IDs a

re li

sted

in ta

ble

1 in

the

mai

n re

port.

USG

S, U

.S. G

eolo

gica

l Sur

vey;

Ass

oc. e

nviro

n. sa

mpl

e, a

ssoc

iate

d en

viro

nmen

tal s

ampl

e; E

. col

i, Es

cher

ichi

a co

li; +

, gen

e pr

esen

t in

sam

ple;

—, g

ene

abse

nt in

sam

ple;

ns,

no sa

mpl

e; V

– re

mar

k co

de in

dica

ting

poss

ible

con

tam

inat

ion

(ana

lyte

was

det

ecte

d in

bot

h th

e en

viro

nmen

tal s

ampl

e an

d th

e as

soci

ated

bla

nks)

]

Shig

ella

Sa

lmon

ella

Ca

mpy

loba

cter

E.

col

i gen

e da

taSa

mpl

e

Ass

oc.

ge

ne d

ata

gene

dat

age

ne d

ata

USG

S

Fiel

d

Flow

co

llect

ion

Sa

mpl

e ty

pe e

nvir

on.

stat

ion

IDID

cond

ition

sda

te s

ampl

erf

b

C. c

oli/

eaeA

stx2

stx1

ipaH

invA

spvC

O15

7C.

jeju

ni

0423

1600

GS-

046/

2/20

11H

igh

Reg

ular

++

——

——

——

0419

3500

MA

-01

3/8/

2011

Hig

hR

egul

ar+

+—

——

++

+04

0854

27M

N-0

27/

6/20

11H

igh

Reg

ular

+—

——

——

——

0408

7170

MW

-01

5/3/

2011

Hig

hR

egul

ar—

——

——

——

—04

0400

00O

N-0

66/

23/2

011

Hig

hR

egul

ar+

——

+—

——

—04

2490

00O

S-01

3/9/

2011

Hig

hR

egul

ar—

——

——

——

—04

2490

00O

S-03

4/27

/201

1H

igh

Reg

ular

—+

——

——

——

0424

9000

OS-

045/

9/20

11H

igh

Reg

ular

——

——

——

——

0424

9000

OS-

159/

7/20

11H

igh

Reg

ular

+—

——

——

——

0419

5500

PO-0

13/

8/20

11H

igh

Reg

ular

+—

——

——

—+

0419

5500

PO-0

24/

20/2

011

Hig

hR

egul

ar+

+—

+—

+—

—04

1955

00PO

-03

7/27

/201

1H

igh

Reg

ular

+—

——

——

——

0419

5500

PO-0

47/

27/2

011

Hig

hR

eplic

ate

PO-0

3+

——

——

——

—

0410

2500

PP-0

24/

21/2

011

Hig

hR

egul

ar+

+—

——

++

+04

1025

00PP

-03

6/1/

2011

Hig

hR

egul

ar+

+—

——

nsns

—04

1025

00PP

-04

7/19

/201

1H

igh

Reg

ular

++

+—

—+

+—

0410

2500

PP-0

58/

3/20

11H

igh

Reg

ular

++

——

——

——

0410

2500

PP-0

810

/4/2

011

Hig

hR

egul

ar+

——

——

——

—04

1665

00R

R-0

15/

16/2

011

Hig

hR

egul

ar+

++

——

—+

—04

1665

00R

R-0

25/

26/2

011

Hig

hR

egul

ar+

++

++

++

—04

1570

00SG

-02

4/6/

2011

Hig

hR

egul

ar+

——

+—

——

—04

1570

00SG

-03

5/2/

2011

Hig

hR

egul

ar+

——

——

——

—04

1570

00SG

-04

5/16

/201

1H

igh

Reg

ular

++

——

——

+—

0410

1500

SJ-0

13/

9/20

11H

igh

Reg

ular

+—

——

——

—+

0410

1500

SJ-0

34/

21/2

011

Hig

hR

egul

ar+

++

+—

+—

—

Appendix 27A

ppen

dix

3.

Path

ogen

gen

e de

tect

ion

in G

reat

Lak

es tr

ibut

arie

s, 2

011.

—Co

ntin

ued

[Stre

amga

ging

stat

ions

cor

resp

ondi

ng to

stat

ion

IDs a

nd fi

eld

IDs a

re li

sted

in ta

ble

1 in

the

mai

n re

port.

USG

S, U

.S. G

eolo

gica

l Sur

vey;

Ass

oc. e

nviro

n. sa

mpl

e, a

ssoc

iate

d en

viro

nmen

tal s

ampl

e; E

. col

i, Es

cher

ichi

a co

li; +

, gen

e pr

esen

t in

sam

ple;

—, g

ene

abse

nt in

sam

ple;

ns,

no sa

mpl

e; V

– re

mar

k co

de in

dica

ting

poss

ible

con

tam

inat

ion

(ana

lyte

was

det

ecte

d in

bot

h th

e en

viro

nmen

tal s

ampl

e an

d th

e as

soci

ated

bla

nks)

]

Shig

ella

Sa

lmon

ella

Ca

mpy

loba

cter

E.

col

i gen

e da

taSa

mpl

e

Ass

oc.

ge

ne d

ata

gene

dat

age

ne d

ata

USG

S

Fiel

d

Flow

co

llect

ion

Sa

mpl

e ty

pe e

nvir

on.

stat

ion

IDID

cond

ition

sda

te s

ampl

erf

b

C. c

oli/

eaeA

stx2

stx1

ipaH

invA

spvC

O15

7C.

jeju

ni

0410

1500

SJ-0

44/

28/2

011

Hig

hR

egul

ar+

—+

——

——

—04

1015

00SJ

-05

6/28

/201

1H

igh

Rep

licat

eSJ

—06

++

——

——

——

0410

1500

SJ-0

66/

28/2

011

Hig

hR

egul

ar+

+—

——

——

—04

2690

00SR

-02

4/6/

2011

Hig

hR

egul

ar+

——

——

——

—04

2690

00SR

-03

4/12

/201

1H

igh

Reg

ular

——

——

——

——

0426

9000

SR-0

55/

5/20

11H

igh

Reg

ular

——

——

——

——

0426

9000

SR-0

65/

24/2

011

Hig

hR

egul

ar—

——

——

——

—04

2690

00SR

-12

8/23

/201

1H

igh

Reg

ular

++

——

+—

+—

0402

4000

ST-0

35/

1/20

11H

igh

Reg

ular

——

——

——

——

0402

4000

ST-1

08/

3/20

11H

igh

Reg

ular

+—

——

——

——


ppen

dix

4.

Qual

ity-c

ontro

l dat

a—Pa

thog

en g

ene

dete

ctio

n in

Gre

at L

akes

trib

utar

ies,

201

1.—

Cont

inue

d

[Stre

amga

ging

stat

ions

cor

resp

ondi

ng to

stat

ion

IDs a

nd fi

eld

IDs a

re li

sted

in ta

ble

1 in

the

mai

n re

port.

USG

S, U

.S. G

eolo

gica

l Sur

vey;

Ass

oc. e

nviro

n. sa

mpl

e, a

ssoc

iate

d en

viro

nmen

tal s

ampl

e;

E. c

oli,

Esch

eric

hia

coli;

+, g

ene

pres

ent i

n sa

mpl

e; -

, gen

e ab

sent

in sa

mpl

e; n

s, no

sam

ple;

v –

val

ue q

ualifi

er c

ode

(ana

lyte

det

ecte

d in

labo

rato

ry b

lank

); V

– v

alue

qua

lifier

cod

e (c

onta

min

atio

n,

anal

yte

was

det

ecte

d in

bot

h th

e en

viro

nmen

tal s

ampl

e an

d th

e as

soci

ated

bla

nks)

]

USGS

stat

ion

IDFi

eld ID

Sam

ple

colle

ctio

n

Date

Sam

ple t

ype

Ass

oc.

e

nviro

n.

sam

ple

E. co

li gen

e dat

aSh

igell

a ge

ne

data

Salm

onell

a ge

ne

data

Cam

pylo

bact

er

gene

da

ta

eaeA

stx2

stx1

rfb O157

ipaH

invA

spvC

C. co

li/C.

jeju

ni04

1375

00A

U-0

88/

23/2

011

Fiel

d bl

ank

AU

-07

——

——

——

——

0402

7000

BD

-06

7/12

/201

1Fi

eld

blan

kB

D-0

4—

——

——

——

—04

0950

90B

N-0

67/

12/2

011

Fiel

d bl

ank

BN

-05

——

——

——

——

0416

5500

CL-

0411

/1/2

011

Fiel

d bl

ank

ns—

——

——

——

—04

2080

00C

Y-04

7/26

/201

1Fi

eld

blan

kC

Y-02

——

——

——

——

0408

5138

5FX

-04

9/19

/201

1Fi

eld

blan

kns

+v—

——

——

——

0411

9400

GR

-07

10/3

1/20

11Fi

eld

blan

kns

——

——

+v—

——

0423

1600

GS-

117/

18/2

011

Fiel

d bl

ank

GS-

09—

——

——

——

—04

1935

00M

A-0

57/

27/2

011

Fiel

d bl

ank

MA

-03

——

——

——

——

0406

7500

MM

-05

9/7/

2011

Fiel

d bl

ank

MM

-03

+V—

—+V

——

——

0408

5427

MN

-05

9/8/

2011

Fiel

d bl

ank

MN

-03

——

——

——

——

0408

7170

MW

-04

9/20

/201

1Fi

eld

blan

kM

W-0

2—

——

——

——

—04

0400

00O

N-1

29/

20/2

011

Fiel

d bl

ank

ns—

——

——

——

—04

2490

00O

S-12

7/18

/201

1Fi

eld

blan

kO

S-10

——

——

——

——

0419

5500

PO-0

57/

27/2

011

Fiel

d bl

ank

PO-0

3—

——

——

——

—04

1665

00R

R-0

510

/31/

2011

Fiel

d bl

ank

ns+v

——

——

——

—04

1765

00R

S-04

8/30

/201

1Fi

eld

blan

kns

——

——

+v—

——

0415

7000

SG-0

98/

23/2

011

Fiel

d bl

ank

SG-0

8—

——

——

——

—04

1015

00SJ

-09

8/24

/201

1Fi

eld

blan

kSJ

-08

——

——

——

——

0426

9000

SR-1

07/

26/2

011

Fiel

d bl

ank

SR-0

8—

——

——

——

—04

0240

00ST

-08

7/21

/201

1Fi

eld

blan

kST

-06

——

——

——

——

0413

7500

AU

-09

8/22

/201

1So

urce

solu

tion

blan

kns

——

——

+v—

——

0402

7000

BD

-07

7/12

/201

1So

urce

solu

tion

blan

kB

D-0

4—

——

——

——

—04

0950

90B

N-0

77/

12/2

011

Sour

ce so

lutio

n bl

ank

BN

-05

——

——

——

——

0416

5500

CL-

0511

/1/2

011

Sour

ce so

lutio

n bl

ank

ns—

——

——

——

—04

2080

00C

Y-05

7/26

/201

1So

urce

solu

tion

blan

kC

Y-02

——

——

——

——

Appendix 29A

ppen

dix

4.

Qual

ity-c

ontro

l dat

a—Pa

thog

en g

ene

dete

ctio

n in

Gre

at L

akes

trib

utar

ies,

201

1.—

Cont

inue

d

[Stre

amga

ging

stat

ions

cor

resp

ondi

ng to

stat

ion

IDs a

nd fi

eld

IDs a

re li

sted

in ta

ble

1 in

the

mai

n re

port.

USG

S, U

.S. G

eolo

gica

l Sur

vey;

Ass

oc. e

nviro

n. sa

mpl

e, a

ssoc

iate

d en

viro

nmen

tal s

ampl

e;

E. c

oli,

Esch

eric

hia

coli;

+, g

ene

pres

ent i

n sa

mpl

e; -

, gen

e ab

sent

in sa

mpl

e; n

s, no

sam

ple;

v –

val

ue q

ualifi

er c

ode

(ana

lyte

det

ecte

d in

labo

rato

ry b

lank

); V

– v

alue

qua

lifier

cod

e (c

onta

min

atio

n,

anal

yte

was

det

ecte

d in

bot

h th

e en

viro

nmen

tal s

ampl

e an

d th

e as

soci

ated

bla

nks)

]

Shig

ella

Salm

onell

a Ca

mpy

loba

cter

E.

coli g

ene d

ata

gene

ge

ne

gene

Sa

mpl

e A

ssoc

.

USGS

stat

ion

data

data

data

Field

IDco

llect

ion

Sa

mpl

e typ

e e

nviro

n.

IDDa

te sa

mpl

erfb

C. co

li/ea

eAst

x2st

x1ip

aHin

vAsp

vCO1

57C.

jeju

ni04

0851

385

FX-0

59/

19/2

011

Sour

ce so

lutio

n bl

ank

ns—

——

——

——

—04

1194

00G

R-0

810

/31/

2011

Sour

ce so

lutio

n bl

ank

ns+v

——

——

——

—04

2316

00G

S-12

7/18

/201

1So

urce

solu

tion

blan

kG

S-09

——

——

——

——

0419

3500

MA

-06

7/27

/201

1So

urce

solu

tion

blan

kM

A-0

3—

——

——

——

—04

0675

00M

M-0

69/

19/2

011

Sour

ce so

lutio

n bl

ank

ns—

——

——

——

—04

0854

27M

N-0

69/

19/2

011

Sour

ce so

lutio

n bl

ank

ns—

——

——

——

—04

0871

70M

W-0

59/

20/2

011

Sour

ce so

lutio

n bl

ank

MW

-02

——

——

——

——

0404

0000

ON

-11

9/20

/201

1So

urce

solu

tion

blan

kns

+v—

——

——

——

0424

9000

OS-

137/

18/2

011

Sour

ce so

lutio

n bl

ank

OS-

10—

——

——

——

—04

1955

00PO

-06

7/27

/201

1So

urce

solu

tion

blan

kPO

-03

——

——

——

——

0416

6500

RR

-06

10/3

1/20

11So

urce

solu

tion

blan

kns

——

——

——

——

0417

6500

RS-

058/

30/2

011

Sour

ce so

lutio

n bl

ank

ns—

——

——

——

—04

1570

00SG

-10

8/23

/201

1So

urce

solu

tion

blan

kSG

-08

——

——

——

——

0410

1500

SJ-1

08/

24/2

011

Sour

ce so

lutio

n bl

ank

SJ-0

8+V

——

——

——

—04

2690

00SR

-11

7/26

/201

1So

urce

solu

tion

blan

kSR

-08

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0402

4000

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21/2

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Brennan and others—Fecal Indicator B

acteria and Gene M

arkers of Pathogenic Bacteria in G

reat Lakes Tributaries, March–O

ctober 2011—Open File Report 2015–1013

ISSN 2331-1258 (online)http://dx.doi.org/10.3133/ofr20151013

occurrence and distribution of fecal indicator …occurrence and distribution of fecal indicator...

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