iii

CONTENTS

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

Introduction ...................................................................................................................................................................... 2

Description of Study Area ................................................................................................................................................ 4

Methods of Study.............................................................................................................................................................. 5

Description of Sampling Network.......................................................................................................................... 5

Sample Collection and Analysis Methods.............................................................................................................. 8

Data Analysis Methods........................................................................................................................................... 12

Assessment of Microbiological Contamination ............................................................................................................... 13

Fecal Indicator Bacteria Densities and Loads ........................................................................................................ 13

Microbial Source Tracking ..................................................................................................................................... 28

Water-Quality Characteristics........................................................................................................................................... 30

Physical Properties ................................................................................................................................................. 30

Dissolved Oxygen......................................................................................................................................... 30

pH ................................................................................................................................................................. 32

Specific Conductance ................................................................................................................................... 33

Water Temperature........................................................................................................................................ 33

Nutrients ................................................................................................................................................................. 33

Dissolved Nitrite Plus Nitrate....................................................................................................................... 34

Total Phosphorus .......................................................................................................................................... 35

Wastewater Organic Compounds............................................................................................................................ 35

Summary........................................................................................................................................................................... 36

References ........................................................................................................................................................................ 41

FIGURES

1. Map showing location of study area..................................................................................................................... 2

2. Photograph showing typical reach of the Jacks Fork immediately upstream from Alley Spring ........................ 4

3. Photograph showing mill at Alley Spring............................................................................................................. 5

4. Graph showing departure from average monthly precipitation at Eminence, Missouri, November 1999

through December 2000 ....................................................................................................................................... 6

5. Photograph showing canoeing on the Jacks Fork................................................................................................. 6

6. Photograph showing trail riders crossing the Jacks Fork ..................................................................................... 8

7. Map showing location of Jacks Fork sites sampled in Phase II............................................................................ 9

8. Graphs showing hydrologic condition during Jacks Fork sampling at two gaged sites, November 1999

through December 2000 ....................................................................................................................................... 11

9. Boxplot example................................................................................................................................................... 15

10. Graphs showing fecal coliform bacteria density at selected Jacks Fork main stem sites and Alley Spring, 

November 1999 through December 2000............................................................................................................. 19

11. Boxplot showing statistical distribution of fecal coliform bacteria density at main stem and tributary

sites and the Eminence Wastewater Treatment Plant on the Jacks Fork, November 1999 through 

December 2000..................................................................................................................................................... 21

12. Graphs showing fecal coliform bacteria density and instantaneous load for Jacks Fork main stem sites

and Alley Spring, November 1999 through December 2000 ............................................................................... 22

13. Graph showing fecal coliform bacteria density at selected Jacks Fork main stem sites and Alley Spring,

July 15 and 16, 2000............................................................................................................................................. 27

14. Pie charts showing percentage of Escherichia coli isolates in water or streambed-sediment samples

assigned to various sources by ribotyping ............................................................................................................ 31
iv 

FIGURES—Continued

15.–17. Boxplots showing:

15. Statistical distribution of dissolved-oxygen concentrations at main stem and tributary sites and

the Eminence Wastewater Treatment Plant on the Jacks Fork, November 1999 through December 

2000 ............................................................................................................................................................. 32

16. Statistical distribution of pH values at main stem and tributary sites and the Eminence Wastewater

Treatment Plant on the Jacks Fork, November 1999 through December 2000 ........................................... 33

17. Statistical distribution of specific conductance values at main stem and tributary sites and

the Eminence Wastewater Treatment Plant on the Jacks Fork, November 1999 through December 

2000 ............................................................................................................................................................. 34

18. Graph showing specific conductance values at Jacks Fork main stem sites and Alley Spring,

November 1999 through December 2000 ............................................................................................................ 35

19. Graph showing water temperature at Jacks Fork main stem sites and Alley Spring, November 1999

through December 2000....................................................................................................................................... 36

20. Boxplot showing statistical distribution of dissolved nitrite plus nitrate concentrations at main stem and 

tributary sites and the Eminence Wastewater Treatment Plant on the Jacks Fork, November 1999 through

December 2000 .................................................................................................................................................... 37

21. Graph showing dissolved nitrite plus nitrate concentrations at Jacks Fork main stem sites and Alley Spring,

November 1999 through December 2000 ............................................................................................................ 38

22. Boxplot showing statistical distribution of total phosphorus concentrations at main stem and tributary

sites and the Eminence Wastewater Treatment Plant on the Jacks Fork, November 1999 through 

December 2000 .................................................................................................................................................... 39

TABLES

1. Estimates of Jacks Fork visitor use for 1999 and 2000...................................................................................... 7

2. Jacks Fork sampling sites, Phase II.................................................................................................................... 10

3. Sample collection dates, hydrologic conditions, and predominant river uses during sampling ........................ 12

4. Analytical methods and method reporting levels............................................................................................... 13

5. Wastewater organic compounds analyzed in selected water samples................................................................ 14

6. Statistical summary of indicator bacteria, physical property, and nutrient data for samples collected 

on the Jacks Fork, tributaries, and the Eminence Wastewater Treatment Plant, November 1999 through 

December 2000 .................................................................................................................................................. 16

7. Fecal coliform and Escherichia coli densities in water and streambed-sediment samples collected at

selected sites, August 2000 ................................................................................................................................ 28

8. Number of Escherichia coli isolates in water or streambed-sediment samples assigned to various 

sources by ribotyping......................................................................................................................................... 29

9. Selected wastewater organic compound concentrations and fecal coliform densities in water samples

collected on July 16 and August 10, 2000 ......................................................................................................... 40
Abstract 1

Assessment of Possible Sources of Microbiological 

Contamination and Water-Quality Characteristics

of the Jacks Fork, Ozark National Scenic Riverways, 

Missouri—Phase II

by Jerri V. Davis and Joseph M. Richards

Abstract

In 1998, an 8-mile reach of the Jacks Fork 

was included on Missouri’s list of impaired waters 

as required by Section 303(d) of the Federal Clean 

Water Act. The identified pollutant on the Jacks 

Fork was fecal coliform bacteria. Potential sources 

of fecal contamination to the Jacks Fork include a 

wastewater treatment plant; campground pit-toilet 

or septic-system effluent; a large commercial, 

cross-country horseback trail riding facility; 

canoeists, boaters, and tubers; and cows.

The U.S. Geological Survey, in cooperation 

with the National Park Service, conducted a study 

to better understand the extent and sources of 

microbiological contamination within the Jacks 

Fork from Alley Spring to the mouth, which 

includes the 8-mile 303(d) reach. Identification of 

the sources would provide the National Park Service and the State of Missouri with the information 

needed to craft a solution of abatement, regulation, 

prevention, and mitigation with the end result 

being the removal of the Jacks Fork from the 

303(d) list. Fifteen sites were sampled from 

November 1999 through December 2000. An 

additional site was sampled one time. Samples 

were collected mostly during base-flow conditions 

during a variety of nonrecreational and recreational season river uses. Samples were analyzed 

for selected fecal indicator bacteria, physical properties, nutrients, and wastewater organic compounds.

During the sampling period, the wholebody-contact recreation standard for fecal 

coliform (200 colonies per 100 milliliters of sample) was exceeded at three sites on August 10, 

2000, and also at one site on May 11, June 7, and 

October 3, 2000. Fecal coliform densities and 

instantaneous loads generally increased from 

background concentrations at the Eminence site, 

peaked about 2 river miles downstream, and then 

decreased until the most downstream site sampled. 

Generally, the largest densities and loads at sites 

downstream from Eminence not related to wetweather flow were observed during a trail ride held 

August 6 to12, 2000. 

A 24-hour sample collection effort was conducted the weekend of July 15 and 16, 2000, to 

investigate the effect that large numbers of swimmers, canoeists, and tubers had on fecal coliform 

densities in the Jacks Fork. Five or six samples 

were collected at six sites between Saturday morning and the following Sunday afternoon. No fecal 

coliform density at any of the sites sampled 

exceeded the whole-body-contact recreation standard.

Because bacteria survive longer in streambed sediments than in water, a source of bacteria in 

the water column could be from resuspension of 

accumulated bacteria from streambed sediments. 

Water and streambed-sediment samples were collected at three sites on August 3, 2000, 1 week 

before a trail ride and again at three sites on 
2 Assessment of Possible Sources of Microbiological Contamination of the Jacks Fork, Missouri—Phase II

August 8, 2000, during a trail ride. Results indicate 

that fecal coliform bacteria densities increased 

substantially in the streambed sediment and the 

water column during the trail ride.

Sixty-five Escherichia coli isolates obtained 

from water samples collected at 9 sites and 23 

Escherichia coli isolates obtained from streambed-sediment samples collected at 5 sites were 

submitted for ribotyping analysis. Samples were 

collected in 2000 during a variety of nonrecreational and recreational season river uses, including trail rides, canoeing, tubing, and swimming. 

Of the 65 isolates from water samples, 40 percent 

were identified as originating from sewage, 29 percent from horse, 11 percent from cow, and 20 percent from an unknown source. Of the 23 isolates 

from streambed-sediment samples, 39 percent 

were identified as originating from sewage, 35 percent from horse, 13 percent from cow, and 13 percent from unknown sources.

Analysis of physical property (dissolved 

oxygen, pH, specific conductance, and temperature) and nutrient (dissolved nitrite plus nitrate and 

total phosphorus) data indicated that overall few 

statistically significant differences occurred 

among the main stem sites of the Jacks Fork. A 

significant increase in total phosphorus concentrations did occur at site 75 immediately downstream 

from the Eminence Wastewater Treatment Plant, 

but the effect diminished quickly downstream. 

Unlike fecal coliform bacteria, most variations in 

physical property values or nutrient concentrations 

were related to seasonal changes, time of day the 

sample was collected, or hydrologic conditions 

and not to certain recreational activities.

Trace quantities of wastewater organic compounds were detected in all waters sampled for 

these constituents. Two of the compounds were 

detected in associated laboratory blanks, and other 

detected compounds have sources other than sewage effluent. The best indicators of municipal or 

domestic sewage effluent were the non-ionic 

detergent metabolites (nonylphenol monoethoxylate, octylphenol monoethoxylate, and para-nonylphenol), phenol, and caffeine; but possible 

sources of these compounds, which were detected 

in one or more of the samples, could be the numerous campers, swimmers, and canoeists that were 

present when the samples were collected.

INTRODUCTION

The Ozark National Scenic Riverways (ONSR), 

the Nation’s first federally protected riverway, was created by an Act of Congress on August 24, 1964, for 

“the purposes of conserving and interpreting unique 

scenic and other natural values and objects of historic 

interest, including preservation of parts of the Current 

River and the Jacks Fork in Missouri as free-flowing 

streams, preservation of springs and caves, management of wildlife, and provisions for use and enjoyment 

of the outdoor recreation resources thereof by the people of the United States” (Public Law 88-492). The primary natural resources protected by the ONSR are 134 

river mi (miles) of the Current River and Jacks Fork 

(fig. 1). About 1.5 million people visit the ONSR annually for recreational purposes, including canoeing, 

johnboating, swimming, fishing, tubing, camping, hiking, caving, horseback riding, and hunting.
Introduction 3

The Jacks Fork is the largest tributary of the Current River, and similar to the Current River, has been 

classified as an Outstanding National Resource Water 

(Missouri Department of Natural Resources, 2000). 

Outstanding National Resource Waters have national 

recreational and ecological significance and receive 

special protection against any degradation in quality. 

The Jacks Fork has been designated for the following 

five beneficial uses by the State of Missouri: livestock 

and wildlife watering, aquatic life protection, coolwater fishery, whole-body-contact recreation, and boating and canoeing (Missouri Department of Natural 

Resources, 2000).

The intense recreational use of the Jacks Fork 

has caused concerns regarding the effects that this use 

might be having on the river. A river use management 

plan prepared by the National Park Service (NPS) (Sullivan and others, 1989) states that the increasing popularity of the recreational area has created concerns 

associated with greater competition for the use of a 

finite resource base. Also, because of inappropriate or 

intensive use, resource damage has increased in some 

areas. Concerns include crowding and increased conflicts between river users, the need to improve and provide more sanitation facilities, the proliferation of litter, 

congestion at river accesses and campgrounds, and balancing the need to protect water quality with the recreational needs of the public.

Section 303(d) of the Federal Clean Water Act 

requires that each State identify those stream segments 

with documented pollution problems for which existing pollution controls are not adequate to implement 

the State water-quality standards. For these impaired 

stream segments, States are required to establish total 

maximum daily loads (TMDL) of the identified pollutant. A TMDL specifies the maximum amount of the 

identified pollutant allowed to be present in a water 

body, allocates allowable pollutant loads among 

sources, and provides the basis for attaining or maintaining water-quality standards within the affected 

water body.

In 1998, an 8 river mi reach of the Jacks Fork was 

included on Missouri’s list of impaired waters as 

required by Section 303(d) of the Federal Clean Water 

Act. The identified pollutant on the Jacks Fork was 

fecal coliform bacteria, whose presence in large numbers indicates contamination by fecal wastes of humans 

and other warm-blooded animals. The State fecal 

coliform standard for safe whole-body-contact recreation is a maximum of 200 col/100 mL (colonies per 

100 milliliters) of sample (Missouri Department of 

Natural Resources, 2000). In the Jacks Fork Basin, the 

standard applies only to the main stem of the Jacks 

Fork during base-flow conditions during the recreational season from April 1 through October 31. Potential sources of fecal contamination to the Jacks Fork 

include the Eminence Wastewater Treatment Plant 

(WWTP); campground pit-toilet or septic-system 

effluent; a large commercial, cross-country horseback 

trail riding facility; canoeists, boaters, and tubers; and 

cows. Studies conducted by the U.S. Geological Survey 

(USGS) (Barks, 1978; Davis and Bell, 1998), Emrie 

(1986), NPS (National Park Service, written commun., 

1997), and the Missouri Department of Natural 

Resources (MDNR) (1998) have indicated that intense 

recreational use is causing adverse affects on the water 

quality of the river, including fecal coliform bacteria 

densities that, on occasion, exceed the water-quality 

standard for whole-body-contact recreation.

Substantive regulatory efforts by the State to 

control and eliminate fecal coliform bacteria inputs to 

the Jacks Fork depend on identification of sources. The 

USGS, in cooperation with the NPS, is conducting a 

three-phase study to better understand the extent and 

sources of microbiological contamination within the 

Jacks Fork from Alley Spring to the mouth (fig. 1), 

which includes the 8 river mi 303(d) reach. The results 

of this study are expected to provide the NPS and other 

natural resource management agencies in Missouri 

with information needed to make effective resourcemanagement decisions. Specific objectives of the 

three-phase study include determining the location and 

magnitude of microbiological contamination (Phase I); 

establishing a water-quality sampling network to further document and understand the sources of microbiological contamination (Phase II); and establishing 

sampling locations for routine long-term water-quality 

monitoring (Phase III).

A summary of the Phase I study is described in 

Davis and Richards (2001). The purpose of this report 

is to describe Phase II of the study. This report includes 

a description of the study area, including geology, climate, land use, and population; a description of the 

sampling network; a description of the sample collection and analysis methods; and an assessment of the 

microbiological contamination and water quality of the 

Jacks Fork using statistical and graphical methods.
4 Assessment of Possible Sources of Microbiological Contamination of the Jacks Fork, Missouri—Phase II

DESCRIPTION OF STUDY AREA

The Jacks Fork is in the Ozark Plateaus physiographic province (Fenneman, 1938) in southeast Missouri, which is an area characterized by deep, narrow 

valleys and sharp ridges (fig. 2). The river flows 

through mature karst topography and gains most of its 

base flow from springs. Karst topography (springs, 

sinkholes, and losing streams) and structural features 

(folds, faults, and fractures) greatly affect water quantity and quality. From its source in Texas County, the 

Jacks Fork drains an area of about 422 mi2 (square 

miles); the average discharge of the Jacks Fork at Eminence, Missouri, is 466 ft3/s (cubic feet per second) 

(Hauck and Nagel, 2000). Alley Spring (fig. 3), Missouri’s seventh largest spring, discharges an average of 

about 125 ft3

/s into the Jacks Fork (Vandike, 1995) 

about 6 river mi upstream from the town of Eminence 

(fig. 1). 

Josiah Bridge mapped and described the geology 

of much of the ONSR area (Barks, 1978). Geologic 

mapping of the Eminence 7-1/2 minute quadrangle was 

completed in 2000, and additional quadrangles are currently (2002) being mapped in the area (R.W. Harrison, 

U.S. Geological Survey, written commun., April 2001). 

The ONSR area is situated on the southwest slope of 

the St. Francois Mountains. Precambrian igneous 

knobs crop out east of Eminence near the center of the 

ONSR. The igneous rocks are mainly rhyolite, a dense, 

fine-grained to porphyritic rock that generally is red but 

may be gray or green. Streams and orifices of large 

springs are in the early Ordovician Gasconade Dolomite and late Cambrian Eminence and Potosi Dolomites. These cherty dolomites generally are several 

hundred feet thick, light gray to brown, and extremely 

soluble. Solution-enlarged openings throughout the 

formations store and transport large quantities of water. 

Soils produced from weathering of the dolomitic rock 

are deep red clays with numerous small chert fragments. The Roubidoux Formation, which is composed 

of interbedded sandstones and dolomites, overlies the 

Gasconade Dolomite and caps the divides between 

most of the streams. Alluvial material in the Jacks Fork 

is comprised of boulders, cobbles, gravel, and sand 

overlying bedrock.

The Jacks Fork area has a temperate climate with 

average annual precipitation of about 42 in/yr (inches 

per year). Average monthly precipitation generally is 

greatest in the spring [March through May; about 4 to 

5 in/mo (inches per month)] and least in the late fall and 

winter (December through February; about 2 to 3 

in/mo; Dugan and Peckenpaugh, 1985). Mean annual 

air temperature is 56 °F (degrees Fahrenheit); the mean 

air temperature during January is 32 °F and during July 

is 78 °F (Dugan and Peckenpaugh, 1985). Monthly precipitation during the sampling period at Eminence, 

Missouri (November 1999 through December 2000), 

generally was below average (fig. 4). 

Land use in the Jacks Fork Basin primarily is forest (76 percent) and agricultural (23 percent). Secondgrowth, deciduous forest mixed with evergreen forest is 

predominant. About 92 percent of the agricultural land 

is used for pasture or hay production (Missouri Spatial 

Data Information Service, 2001). The permanent population in the Jacks Fork Basin is about 8,000; the largest towns are Eminence (548) and Mountain View 

(2,430) (University of Missouri, 2001). Estimates of 

monthly horseback trail riders, canoe (fig. 5), tube, and 

angler use for 1999 and 2000 were compiled by the 

NPS as part of the ONSR rangers’ monthly visitor use 

reports (table 1). During the Phase II sampling period, 

six guided week-long trail rides were held on May 7 to 
Methods of Study 5

13, June 4 to 10, August 6 to 12, October 1 to 7, 

November 5 to 11, and December 3 to 8, 2000. The 

number of trail riders (fig. 6) is based only on guided 

trail rides inside the ONSR and does not include 

unguided rides in the ONSR. This estimate is, therefore, substantially less than the actual number of trail 

riders that participate in any of the week-long trail 

rides. During the largest trail rides held in June, 

August, and October 2000, an estimated 3,000 total 

trail riders may have taken advantage of the recreational opportunities on the Jacks Fork (Dr. Kenneth 

Chilman, Southern Illinois University at Carbondale, 

written commun., 2001). 

METHODS OF STUDY

To determine the microbiological contamination 

and water quality of the study area, a network of sampling sites was established. Water samples were collected from these sites and analyzed for a variety of 

indicator bacteria and chemical constituents.

Description of Sampling Network

The locations of microbiological contamination were 

determined in Phase I through three intensive synoptic 

surveys (Davis and Richards, 2001). A synoptic survey 
6 Assessment of Possible Sources of Microbiological Contamination of the Jacks Fork, Missouri—Phase II

consists of the measurement of selected constituents at 

many sites during a brief period representative of a particular hydrologic or seasonal condition. A reconnaissance of the Jacks Fork from the Alley Spring 

Campground to the mouth was done to locate sampling 

sites, locate potential sources of microbiological contamination, and map hydrologic features for inclusion in 

a geographic information system (GIS) database. Each 

location or feature was identified by geographic coordinates as determined by a global positioning system 

(GPS). Based on information collected during the reconnaissance, 42 sampling sites, including the Eminence 

WWTP, were selected (Davis and Richards, 2001). 

These sites were located on tributaries, spring branches, 

and on the main stem of the Jacks Fork near potential 

sources of microbiological contamination. The first 

intensive synoptic survey was done May 10 to 12, 1999, 

during spring base-flow conditions at the beginning of 

the recreational season (defined by the State of Missouri 

as April 1 through October 31). The second and third 

intensive synoptic surveys were done during the recreational season on June 22 to 24 and August 10 to 12, 

1999, during early-summer and late-summer base-flow 

conditions.
Methods of Study 7

Table 1. Estimates of Jacks Fork visitor use for 1999 and 2000

Month

Horseback 

ridersa Canoesb Tubesb Anglersc

1999d

January 137 0 0 119

February 0 20 0 82

March 6 47 0 104

April 41 186 0 233

May 125 665 5 729

June 65 3,326 497 986

July 224 4,302 387 1,786

August 296 3,760 308 382

September 49 1,079 98 453

October 403 222 0 398

November 142 12 0 282

December 25 4 0 154

Total 1,513 13,623 1,295 5,708

2000d

January 31 8 0 209

February 17 20 0 115

March 12 50 0 150

April 213 225 0 457

May 387 828 5 1,176

June 787 3,513 114 1,454

July 383 4,561 566 760

August 283 4,075 363 1,892

September 254 1,665 91 558

October 642 150 0 428

November 126 29 0 286

December 16 20 0 43

Total 3,151 15,144 1,139 7,528

a The number of horseback riders is derived from both the cross-country trail riding facility and National Park 

Service rangers during their normal duties in the area. The cross-country trail riding facility does not report all 

riders participating in a week-long trail ride. Only guided trail rides inside the Ozark National Scenic Riverways 

are reported; unguided trail riders in the Ozark National Scenic Riverways are not counted by the cross-country 

trail riding facility. b The number of canoes and tubes reflects the number of canoes and tubes reported as rented by concessionaires in the area. c The number of anglers reflects the number of anglers observed by National Park Service rangers during their 

normal duties in the area. d These numbers are totals for the entire Jacks Fork within the boundaries of the Ozark National Scenic Riverways derived from the National Park Service rangers’ monthly visitor use reports.
8 Assessment of Possible Sources of Microbiological Contamination of the Jacks Fork, Missouri—Phase II

Phase I results indicated that fecal coliform bacteria are a concern in the Jacks Fork (Davis and Richards, 2001). Fecal coliform bacteria densities and loads 

generally were larger in the 303(d) reach downstream 

from Eminence than upstream from Eminence during 

the June 22 to 24 and August 10 to 12, 1999, synoptic 

surveys. The State standard for whole-body-contact 

recreation was exceeded at two sites in the 303(d) reach 

during the June synoptic survey and at five sites in the 

303(d) reach during the August synoptic survey.

To further understand the sources of microbiological contamination in the Jacks Fork, sampling was 

done during Phase II (November 1999 through December 2000) at 16 of the 42 sites from Phase I (fig. 7, table 

2). An additional site not sampled during Phase I was 

sampled one time. Samples were collected mostly during base-flow conditions during a variety of nonrecreational and recreational season river uses, including 

canoeing, swimming, tubing, camping, and horseback 

riding. River conditions were above base flow when 

samples were collected during December 1999 and late 

February and early March 2000. Continuously recording streamflow gaging stations are located on the Jacks 

Fork immediately downstream from site 5 and at Eminence (site 60; fig. 7). The hydrologic conditions at the 

two gaged sites and river uses occurring during Phase II 

sampling are shown in figure 8 and listed in table 3. 

Although the recreational season is defined as occurring 

between April 1 and October 31, most recreational river 

use takes place between late May and early September.

Sample Collection and Analysis Methods

During Phase II, 15 sites were sampled approximately monthly from November 1999 through April 

2000 and from September 2000 through December 

2000 and twice per month from May 2000 through 

August 2000 (table 3). Samples were collected and 

analyzed by the USGS at each site for indicator bacteria 

[fecal coliform, Escherichia coli (E. coli), fecal streptococci, and enterococci] using the membrane filtration 

procedure described in Myers and Wilde (1997). Indicator bacteria samples were collected in a sterile 500-

mL (milliliter) polypropylene bottle by facing the bottle into the current and dipping quickly into the stream 

at 3 to 5 equally spaced locations in the stream cross 

section. The samples were placed on ice and held a 

maximum of 6 hours until processing. Because densities of indicator bacteria can be quite variable, generally two sample volumes ranging from 10 to 100 mL 

were filtered from individual stream samples. Reagent 

blanks were run twice each day to check for contamination of equipment and reagents.

Samples for the enumeration of fecal indicator 

bacteria (fecal coliform and E. coli) in streambed sediments were collected at selected sites in August 2000 

using a modification of the membrane filtration procedure described in Francy and Darner (1998). Streambed sediments, which consist primarily of gravel, sand, 

and silt, were collected by scooping the stream bottom 

with a sterile wide-mouth 250-mL polypropylene jar. 

The jars were kept covered until they touched the 

stream bottom to minimize contamination by the overlying water and then quickly closed after scooping the 

sediment. Streambed sediments were subsampled in 

three locations in the stream cross section. The samples 

were placed on ice and held no more than 24 hours 

before processing. The three subsamples were composited in a sterile 1-L (liter) polypropylene bottle and then 

split into two sterile 500-mL polypropylene jars. Phosphate buffer was added to one of the subsamples to isolate fecal coliform, and saline buffer was added to the 

other subsample to isolate E. coli. The samples were 

shaken vigorously for 5 minutes, the buffer-sediment 

mixture decanted into two 250-mL polypropylene bottles, and the sediment allowed to settle. Three sample 

volumes ranging from 1 to 10 mL were filtered from 

the individual streambed-sediment sample extracts for 

analysis of fecal coliform and E. coli.

A technique known as Microbial Source Tracking (MST) was used in this study to help identify the 

primary sources of E. coli in water and streambed-sediment samples. Ribotyping, which can be used for 

MST, has been shown to be useful in discriminating 

between human and nonhuman sources of E. coli in 

water samples (Hartel and others, 1999; Parveen and
Sample Collection and Analysis Methods 9 0 1 2 3 MILES 0 1 2 3 KILOMETERS

106

19

V

EMINENCE

Circle B

Campground

Alley

Spring

Alley Spring

Campground

Jacks Fork

Campground

Cross-country

trail riding

facility

Shawnee Creek

Campground

15

5

35

45

60 70

75 80

110

140

150 160

185

170

28*

165

37°11"30'

37°09'

91°25' 91°22'30" 91°20' 91°17'30"

Jacks

Fork

Storys

Creek

Creek

Shawnee

Creek

Creek

Little

Shawnee

Current

River

Mahans

EXPLANATION

Base from U.S. Geological Survey digital data, 1:100,000, 1987Universal Transverse Mercator projectionZone 15

Figure 7. Location of Jacks Fork sites sampled in Phase II.

OZARK NATIONAL SCENIC RIVERWAYS303(d) REACH

45 SAMPLING SITE AND IDENTIFIER—asterisk indicates site was sampled July 15 and 16, 2000, only

SAMPLING SITE AND GAGING STATION AND IDENTIFIEREMINENCE WASTEWATER TREATMENT PLANT AND IDENTIFIER

6070

Effluent

pipe
10 Assessment of Possible Sources of Microbiological Contamination of the Jacks Fork, Missouri—Phase II

others, 1999; Schlottmann and others, 2000; Carson 

and others, 2001). Ribotyping involves isolating pure 

cultures of E. coli from both an environmental sample 

and known-source feces. The deoxyribonucleic acid 

(DNA) is isolated from these pure bacterial strains, cut 

in fragments using a restriction enzyme, and the resulting fragments are separated by molecular weight using 

agarose-gel electrophoresis. Hybridization with a 

labeled DNA probe creates a chemiluminescent pattern 

of the fragments containing ribosomal ribonucleic acid 

(rRNA) genetic information. The resulting banding 

patterns from water-isolated E. coli are compared to the 

banding patterns of isolated E. coli from known-source 

feces for similarity and possible identification. Water 

samples for ribotyping analysis were collected from 

selected sites in April, June, July, and August 2000 

using the same technique described previously for indicator bacteria and sent by overnight mail to the College 

of Veterinary Medicine at the University of Missouri in 

Columbia (UMC), Missouri. For this study, the 

unknowns were compared to the banding patterns from 

sewage, horses, and cows.

Samples for nutrient analyses (dissolved nitrite 

plus nitrate, ammonia, and orthophosphate and total 

ammonia plus organic nitrogen and phosphorus) also 

were collected at the 15 Phase II sites according to standard USGS sample collection and processing protocols 

described by Edwards and Glysson (1998) and Wilde 

and others (1999a, 1999b). All chemical analyses were 

done by the USGS National Water Quality Laboratory 

in Lakewood, Colorado, according to procedures 

described in Fishman and Friedman (1989) or Fishman 

(1993) (table 4). Selected sites (sites 5, 15, 60, 80, and 

110; fig. 7; table 2) also were sampled for wastewater 

organic compounds (table 5) on July 16 and August 10, 

2000 (Zaugg and others, 2002). Onsite measurements 

of dissolved oxygen (DO), pH, specific conductance, 

and temperature were made at each site according to 

procedures described by Wilde and Radtke (1998). 

a

 Recharge area of Alley Spring is estimated to be greater than 125 square miles (Aley and Aley, 1987). b This site was sampled only during the weekend of July 15 and 16, 2000.

Table 2. Jacks Fork sampling sites, Phase II

[mi2, square miles; NA, not applicable]

Site 

number 

(fig. 7) Site name Latitude Longitude

Drainage 

area 

(mi2)

5 Jacks Fork above Alley Spring 370857 912659 302

15 Alley Spring at Alley 370914 912629 a

28 Jacks Fork above Eminenceb 370931 912341 340

35 Mahans Creek above Eminence 370911 912232 54.1

45 Storys Creek above Eminence 370928 912208 5.34

60 Jacks Fork at Eminence 370918 912131 404

70 Eminence Wastewater Treatment Plant 370915 912101 NA

75 Jacks Fork below Wastewater Treatment Plant below Eminence 370907 912108 405

80 Jacks Fork above 2nd unnamed hollow (south) below Eminence 370905 912040 406

110 Jacks Fork above Lick Log Hollow below Eminence 371014 912013 409

140 Jacks Fork above Bald Knob Hollow below Eminence 371012 911913 412

150 Jacks Fork above Powell Spring above Two Rivers 371026 911833 413

160 Shawnee Creek above Two Rivers 371019 911801 20.0

165 Jacks Fork above Little Shawnee Creek above Two Rivers 371020 911741 433

170 Little Shawnee Creek above Two Rivers 371021 911732 9.4

185 Jacks Fork below 3rd unnamed hollow (north) above Two Rivers 371054 911735 444
Sample Collection and Analysis Methods 11
12 Assessment of Possible Sources of Microbiological Contamination of the Jacks Fork, Missouri—Phase II

Stream discharge was measured at all tributary sites 

and at most of the other Jacks Fork main stem sites during sample collection. 

Twenty-two quality-assurance samples were collected at the 15 sites. The quality-assurance samples 

included 8 field equipment blanks (FEB) collected to 

monitor for contamination and carry-over between 

environmental samples and 14 replicate environmental 

samples collected to monitor analytical precision. 

Nutrients were analyzed in the FEB and replicate samples. Most constituent concentrations were less than the 

method reporting level (MRL) in the 8 FEB with the 

following exceptions: total ammonia plus organic 

nitrogen detected in 4 FEB [MRL 0.10 mg/L (milligram per liter), and detected concentrations ranged 

from an estimated concentration of 0.06 to 0.18 mg/L]; 

and dissolved orthophosphate detected in 1 FEB (MRL 

0.01 mg/L, detected concentration of 0.012 mg/L). 

Nutrient concentrations in the replicate environmental 

samples were comparable and within laboratory analytical error.

Data Analysis Methods

The indicator bacteria, physical property, and 

nutrient data were evaluated to determine factors 

affecting their occurrence in the Jacks Fork and tributaries. Descriptive statistics were used to show the central tendency and variation in the data. The minimum 

and maximum and the values at the 25th, 50th 

(median), and 75th percentiles were calculated. The 

water-quality data collected during Phase II are available in Hauck and Nagel (2001).

Table 3. Sample collection dates, hydrologic conditions, and predominant river uses during sampling

Date of sample collection Hydrologic condition River use

November 8–10, 1999 Base flow Nonrecreational season 

(trail ride October 31–November 6)

December 14–16, 1999 Falling stage Nonrecreational season

January 18–20, 2000 Base flow Nonrecreational season

February 29–March 2, 2000 Falling stage Nonrecreational season

April 4–6, 2000 Base flow Early recreational season

May 10–12, 2000 Base flow Trail ride, canoeing

May 23–25, 2000 Base flowa

a

 Base-flow conditions existed on May 23 and 24. Rainfall on the evening of May 24 and the early morning of May 25 resulted 

in runoff. The whole-body-contact recreation standard does not apply during wet-weather flow.

Canoeing, swimming, tubing, and camping

June 6–8, 2000 Base flow Trail ride, canoeing, swimming, tubing, and camping

June 28–30, 2000 Base flow Canoeing, swimming, tubing, and camping

July 10–12, 2000 Base flow Canoeing, swimming, tubing, and camping

July 15–16, 2000 Base flow Canoeing, swimming, tubing, and camping

July 26–28, 2000 Base flow Canoeing, swimming, tubing, and camping

August 2, 2000 Base flow Canoeing, swimming, tubing, and camping

August 8–11, 2000 Base flow Trail ride, canoeing, swimming, tubing, and camping

August 21–22, 2000 Base flow Canoeing, swimming, tubing, and camping

September 19–20, 2000 Base flow Late recreational season

October 3–4, 2000 Base flow Trail ride and late recreational season

November 8–9, 2000 Base flow Trail ride and nonrecreational season

December 12 and 20, 2000 Base flow Nonrecreational season
Assessment of Microbiological Contamination 13

The distribution of selected indicator bacteria, 

physical property, and nutrient constituent data was 

graphically displayed using side-by-side boxplots (fig. 

9; Helsel and Hirsch, 1992, p. 24–26). The plots show 

the center of the data (median—the center line of the 

box), the variation (interquartile range—the box 

height), the skewness (quartile skew—the relative size 

of the box halves), and the presence or absence of 

unusual values (“outside” or “far-out” values). The 

boxplot consists of a center line (the median) splitting 

a rectangle defined by the interquartile range (25th to 

75th percentiles) and whiskers that extend to the last 

observation within one step beyond either end of the 

box (“adjacent values”). A step equals 1.5 times the 

height of the box. Observations between one and two 

steps from the box in either direction are plotted with 

an asterisk (“outside values”), and observations farther 

than two steps beyond the box are plotted with a circle 

(“far-out values”). If the median equals the 25th percentile, no center line will be present. If the median 

equals the 25th and 75th percentiles, the box is represented by a line. Boxplots constructed for sites with 

censored data (data reported as less than some threshold) were modified by making the lower limit of the 

box equal to the MRL.

The nonparametric Kruskal-Wallis analysis-ofvariance test (Helsel and Hirsch, 1992, p. 163) was 

used to test for differences in the distributions of the 

data from Alley Spring and the nine Jacks Fork main 

stem sampling sites. The distributions were considered 

significantly different from one another if the probability (p-value) was less than 5 percent (less than 0.05) 

that the observed difference occurred by chance. If a 

statistically significant difference was detected 

between the sites, individual differences were evaluated by applying Tukey’s multiple comparison test to 

the rank-transformed data (Helsel and Hirsch, 1992, p. 

196). 

ASSESSMENT OF MICROBIOLOGICAL 

CONTAMINATION

To determine the probable sources of fecal bacteria, multiple lines of evidence are necessary, including microbiological, physical, chemical, and 

hydrological data and ancillary information on land, 

water, and recreational use of the resource. The indicator bacteria analyzed for this study include fecal 

coliform, E. coli, fecal streptococci, and enterococci. 

Summary statistics of the indicator-bacteria data for 

each of the 15 sites are listed in table 6. A complete 

analysis of the fecal coliform data is presented in the 

following sections. Results for E. coli, fecal streptococci, and enterococci are similar to those for fecal 

coliform and will not be discussed.

Fecal Indicator Bacteria Densities and 

Loads

Fecal indicator bacteria are measures of the sanitary quality of water. The density of these bacteria is 

one indicator of whether water is safe for whole-bodycontact recreation and consumption and/or free from 

disease-causing organisms. Indicator bacteria are not 

typically disease causing, but two indicator organisms, 

E. coli and enterococci, have been strongly correlated

Table 4. Analytical methods and method reporting levels

[mg/L, milligram per liter; N, nitrogen; ASF, automated-segmented flow; P, phosphorus; EPA, U.S. Environmental Protection Agency] 

Analyte Analytical methoda

a

Fishman, 1993 and Fishman and Friedman, 1989.

Method number

Method reporting level

(mg/L)

Nitrite plus nitrate, dissolved as N Colorimetry, ASF I-2545-90 0.05

Ammonia, dissolved as N Colorimetry, ASF I-2522-90 .02

Ammonia plus organic nitrogen, 

total as N

Colorimetry, ASF, 

microkjeldahl digestion

I-4515-91 .10

Phosphorus, total as P Colorimetry, ASF, 

microkjeldahl digestion

EPA 365.1 .004

Orthophosphate, dissolved as P Colorimetry, ASF I-2601-90 .01
Table 5. Wastewater organic compounds analyzed in selected water samples

[All compounds analyzed by gas chromatography/mass spectrometry using method O-1433-01 (Zaugg and others, 2002); μg/L, micrograms per liter]

Compound

Method

reporting level

(μg/L) Compound

Method

reporting level

(μg/L)

Non-ionic detergent metabolites Stimulants and metabolites

NPEO1 (nonylphenol monoethoxylate) 1.00 Caffeine 0.08

NPEO2 (nonylphenol diethoxylate) 1.10 Codeine (analgesic) .20

OPEO1 (octylphenol monoethoxylate) .10 Cotinine (nicotine metabolite) .04

OPEO2 (octylphenol diethoxylate) .20

para-Nonylphenol (total) .50 Plasticizers and polymer precursors

bis(2-Ethylhexyl) adipate 2.00

Disinfectants Bisphenol A (polymer manufacture) .09

Phenol .25 bis(2-Ethylhexyl) phthalate 2.50

Triclosan (antimicrobial) .05 Diethylphthalate .25

Ethanol, 2-butoxy-, phosphate .20

Fecal indicator and hormones Phthalic anhydride (plastic manufacture) .29

3-beta-Coprostanol (carnivores) .60 Triphenyl phosphate .10

Cholesterol 1.50

17-beta-Estradiol (estrogen metabolite) .50 Polynuclear Aromatic Hydrocarbons (PAHs)

Stigmastanol (plant sterol) 2.00 Anthracene .06

Benzo[a]pyrene .05

Fire retardants Fluoranthene .03

Tri(2-chloroethyl) phosphate .04 Naphthalene .02

Tri(dichloroisopropyl) phosphate .10 Phenanthrene .05

Pyrene .03

Fragrance

Acetophenone .22 Food and other preservatives

5-Methyl-1H-benzotriazole (industrial use) .10

Fumigant 2,6-Di-tert-butylphenol .08

1,4-Dichlorobenzene .03 2,6-Di-tert-para-benzoquinone .50

Butylated hydroxyanisole (BHA) .12

Pesticides Butylated hydroxytoluene (BHT) .08

Carbaryl .06 para-Cresol (wood preservative) .06

Chlorpyrifos .02

cis-Chlordane .04 Solvent

Dieldrin .08 Tetrachloroethylene .03

Diazinon .03

Lindane .05

Methyl parathion .06

N,N-diethyltoluamide (DEET) .04

14 Assessment of Possible Sources of Microbiological Contamination of the Jacks Fork, Missouri—Phase II
Assessment of Microbiological Contamination 15

to the presence of water-borne pathogens (U.S. Environmental Protection Agency, 1986). Common sources 

of fecal indicator bacteria include municipal wastewater-treatment effluents that have not been disinfected; 

septic tanks; animal wastes from feedlots, barnyards, 

and pastures; and stormwater. The fecal indicator bacteria species used in the study are bacteria of the fecal 

coliform and fecal streptococci groups, E. coli, and 

enterococci; all of which commonly are found in the 

intestinal tracts of warm-blooded animals. E. coli and 

enterococci are strictly inhabitants of the gastrointestinal tracts of warm-blooded animals, and their presence 

in water is direct evidence of fecal contamination from 

warm-blooded animals and the possible presence of 

pathogens (Dufour, 1977).

Because fecal bacteria normally inhabit the gastrointestinal tracts of warm-blooded animals, they die 

off within days of exposure to the relatively cold environment of streams. Fecal bacteria have been shown to 

survive from a few hours to a few days in streams and 

a few weeks to a few months in streambed sediments 

(Roszak and Colwell, 1987; Doyle and others, 1992; 

Pommequy and others, 1992; Myers and others, 1998). 

Bacterial decay results from cell starvation, predation 

by stream protozoans, and destruction by the ultraviolet 

component of sunlight and other physicochemical processes such as chlorination (U.S. Environmental Protection Agency, 1985; Roszak and Colwell, 1987).

During the sampling period, the whole-bodycontact recreation standard (200 col/100mL)—applicable from April 1 through October 31 during base-flow 

conditions on the main stem of the Jacks Fork—was 

exceeded at one or more sites on four sampling occasions. The standard was exceeded at sites 80, 110, and 

140 on August 10, 2000 (210, 340, and 210 col/100 mL 

of sample) and also at site 110 on May 11 (340 col/100 

mL), June 7 (240 col/100 mL), and October 3, 2000 

(780 col/100 mL) (fig. 10; site 140 is not included on 

the figure because fewer samples were collected at site 

140 than at the other sites shown on the graph). Each of 

these exceedences occurred in samples collected during trail rides (table 3). Values above the standard also 

were measured in samples collected on May 25, 2000, 

at sites 165 and 185 (450 and 490 col/100 mL), but 

these probably occurred as a result of runoff during 

wet-weather flow when the standard does not apply 

(fig. 8). 

The distribution of fecal coliform bacteria densities at nine main stem sites, five tributary sites, and the 

Eminence WWTP is shown in figure 11. The only statistically significant difference (p value less than 0.05)
16 Assessment of Possible Sources of Microbiological Contamination of the Jacks Fork, Missouri—Phase II Table 6. Statistical summary of indicator bacteria, physical property, and nutrient data for samples collected on the Jacks Fork, tributaries, and the Emine

November 1999 through December 2000 [col/100 mL, colonies per 100 milliliters; E. coli, Escherichia coli; ft3/s, cubic feet per second; mg/L milligrams per liter; μS/cm, microsiemens per centimeter at 25 degrees Celsius; °C, degrees 

than; --, no data available] Fecal coliform (col/100 mL) E. coli (col/100 mL) Fecal streptococci (col/100 mL) Enterococci (col/100 mL) Discharge (ft3/s) Dissolved oxygen (mg/L) pH Specific conductance

(μS/cm)

Water 

temperature

(°C)

Nitrite plus 

nitrate, 

dissolved 

(mg/L as N)

Ammonia, 

dissolved 

(mg/L as N)

p

(

Jacks Fork above Alley Spring (site 5, fig. 7)

Number of samples 18 18 18 14 18 18 18 18 18 18 18 Minimum <1 <1 <1 3 25 5.8 7.2 246 1.0 <.05 <.02 25th percentile 2 1 14 8 42 8.5 7.9 311 11.7 .09 <.02 Median 8 3 40 18 57 9.9 8.1 332 21.9 .11 <.02 75th percentile 23 12 67 32 117 10.7 8.2 337 25.5 .22 <.02 Maximum 64 55 180 236 359 15.6 8.4 346 29.5 .68 <.02 

Alley Spring at Alley (site 15, fig. 7)

Number of samples 18 18 18 14 18 18 18 18 18 18 18 Minimum 2 1 4 6 66 7.2 7.5 244 13.6 .53 <.02 25th percentile 6 2 27 27 74 9.6 7.7 296 14.0 .56 <.02 Median 10 8 41 38 80 10.2 7.7 310 14.6 .58 <.02 75th percentile 18 17 50 52 87 10.7 7.8 316 14.9 .63 <.02 Maximum 92 56 84 124 173 11.3 7.9 322 15.3 .77 <.02 

Mahans Creek above Eminence (site 35, fig. 7)

Number of samples 17 17 17 13 17 17 17 17 17 12 12 Minimum 16 2 24 50 2.8 7.9 7.9 319 3.2 .16 <.02 25th percentile 32 20 94 98 5.4 9.0 8.1 388 11.4 .19 <.02 Median 96 48 130 160 8.5 9.5 8.1 411 19.6 .20 <.02 75th percentile 180 110 220 220 12 10.0 8.1 435 21.9 .26 <.02 Maximum 440 280 430 1,270 58 14.2 8.3 458 25.9 .32 <.02 

Storys Creek above Eminence (site 45, fig. 7)

Number of samples 10 10 10 6 10 10 10 10 10 5 5 Minimum 27 16 30 24 .30 6.9 8.0 367 5.7 .15 <.02 25th percentile 50 31 60 410 .52 8.0 8.2 427 10.2 -- -- Median 85 64 295 555 .69 9.2 8.3 452 17.8 .19 <.02 75th percentile 540 320 630 670 1.2 10.2 8.4 466 23.6 -- -- Maximum 1,500 680 1,200 1,200 4.6 13.4 8.5 511 28.4 .28 <.02 

Jacks Fork at Eminence (site 60, fig. 7)

Number of samples 21 21 21 16 21 21 20 21 21 18 18 Minimum 2 <1 4 <1 110 7.9 7.8 263 4.2 .21 <.02 25th percentile 5 3 15 14 136 9.2 8.0 320 13.0 .28 <.02 Median 25 13 40 27 154 9.9 8.1 327 17.9 .30 <.02 75th percentile 35 21 92 62 224 11.2 8.3 331 20.2 .38 <.02 Maximum 120 86 230 170 542 15.1 8.5 338 24.7 .66 <.02 
Assessment of Microbiological Contamination 17

Table 6. Statistical summary of indicator bacteria, physical property, and nutrient data for samples collected on the Jacks Fork, tributaries, and the Eminence November 1999 through December 2000–Continued

Fecal 

coliform 

(col/100 mL)

E. coli

(col/100 mL)

Fecal 

streptococci 

(col/100 mL)

Enterococci 

(col/100 mL)

Discharge

(ft3/s)

Dissolved 

oxygen

(mg/L) pH

Specific 

conductance

(μS/cm)

Water 

temperature

(°C)

Nitrite plus 

nitrate, 

dissolved 

(mg/L as N)

Ammonia, 

dissolved 

(mg/L as N)

Amm

plus o

N, t

(mg/L 

Eminence Wastewater Treatment Plant (site 70, fig. 7)

Number of samples 13 13 13 9 -- 6 6 12 12 9 9 9 Minimum <1 <1 <1 <1 -- 1.1 7.7 509 9.8 .07 <.02 25th percentile <1 <1 <1 <1 -- 2.2 7.7 608 11.6 7.04 <.02 Median <1 <1 <1 <1 -- 3.6 7.7 752 21.2 16.2 <.02 75th percentile <1 <1 2 5 -- 5.3 7.8 805 24.0 19.8 .36 1Maximum 5 10 21 11 -- 5.6 7.8 1040 27.0 24.8 8.2 10 

Jacks Fork below Wastewater Treatment Plant below Eminence (site 75, fig. 7)

Number of samples 14 14 14 10 -- 14 14 14 14 14 14 14 Minimum 2 1 5 5 -- 8.6 7.9 265 8.9 .29 <.02 <25th percentile 11 6 14 16 -- 9.2 8.0 317 13.4 .36 <.02 Median 22 18 44 30 -- 10.2 8.1 328 18.0 .38 <.02 75th percentile 48 24 72 45 -- 11.0 8.2 331 21.3 .42 <.02 Maximum 200 180 300 120 -- 12.4 8.4 338 25.3 .67 .02 

Jacks Fork above 2nd unnamed hollow below Eminence (site 80, fig. 7)

Number of samples 17 17 17 13 17 17 17 17 17 17 17 17 Minimum 4 <1 3 5 109 8.9 8.0 265 6.7 .22 <.02 <25th percentile 10 3 14 13 128 9.3 8.1 323 13.1 .30 <.02 <Median 17 7 34 18 155 10.2 8.2 330 18.1 .34 <.02 75th percentile 48 41 70 69 228 11.5 8.3 333 19.6 .40 <.02 Maximum 210 220 700 190 547 14.0 8.5 338 24.2 .66 <.02 

Jacks Fork above Lick Log Hollow below Eminence (site 110, fig. 7)

Number of samples 19 19 19 15 17 19 19 19 19 18 18 17 Minimum <1 <1 4 1 105 7.4 7.8 267 3.3 .22 <.02 <25th percentile 18 6 21 10 133 9.4 8.2 321 14.0 .30 <.02 Median 50 29 48 60 151 10.4 8.3 331 19.2 .31 <.02 75th percentile 130 82 160 180 227 11.2 8.4 338 21.5 .36 <.02 Maximum 780 210 710 1,080 524 14.9 8.5 340 24.4 .64 <.02 

Jacks Fork above Bald Knob Hollow below Eminence (site 140, fig. 7)

Number of samples 12 12 12 10 -- 11 12 12 11 12 12 12 Minimum 1 <1 4 2 -- 7.6 8.1 301 9.0 .16 <.02 <25th percentile 12 4 16 8 -- 9.6 8.1 322 17.9 .27 <.02 Median 23 11 36 30 -- 10.7 8.3 331 21.8 .28 <.02 75th percentile 33 16 88 62 -- 11.1 8.4 338 22.5 .33 <.02 Maximum 210 48 210 130 -- 12.6 8.5 344 25.7 .40 .02 
18 Assessment of Possible Sources of Microbiological Contamination of the Jacks Fork, Missouri—Phase II Jacks Fork above Powell Spring above Two Rivers (site 150, fig. 7) Number of samples 17 17 17 13 16 17 17 17 17 17 17 17 

Minimum 1 <1 6 3 114 6.7 7.4 273 2.6 .21 <.02 <25th percentile 15 2 13 8 135 9.1 8.1 324 12.9 .26 <.02 Median 33 15 28 14 160 10.7 8.3 333 20.2 .28 <.02 75th percentile 49 23 46 55 218 11.2 8.4 340 22.5 .35 <.02 Maximum 100 66 120 220 489 14.5 8.5 346 26.1 .67 <.02 

Shawnee Creek above Two Rivers (site 160, fig. 7) Number of samples 17 17 17 13 17 17 17 17 17 12 12 12 

Minimum <1 2 7 8 2.3 6.0 7.9 338 4.1 .10 <.02 <25th percentile 18 13 70 120 3.9 8.7 8.1 458 12.3 .19 <.02 <Median 72 40 160 170 5.7 9.3 8.1 476 19.6 .23 <.02 75th percentile 120 82 220 490 11 11.5 8.2 501 20.1 .35 <.02 Maximum 600 420 1,500 1,000 24 14.5 8.4 526 26.2 .59 <.02 

Jacks Fork above Little Shawnee Creek above Two Rivers (site 165, fig. 7) Number of samples 17 17 17 13 -- 17 17 17 17 17 17 17 

Minimum 3 <1 5 5 -- 6.8 7.9 284 5.5 .21 <.02 <25th percentile 10 7 12 18 -- 9.5 8.2 329 12.8 .25 <.02 <Median 50 18 50 61 -- 10.4 8.2 335 19.4 .30 <.02 75th percentile 69 34 84 120 -- 11.4 8.3 340 20.7 .37 <.02 Maximum 450 570 4,100 3,100 -- 14.2 8.4 347 27.0 .65 <.02 

Little Shawnee Creek above Two Rivers (site 170, fig. 7) Number of samples 8 8 8 5 8 8 8 8 8 4 4 4 

Minimum 12 2 23 25 .56 5.3 7.8 414 6.5 .17 <.02 <25th percentile 13 12 30 86 .68 8.0 8.0 449 8.6 .25 -- -Median 18 16 100 97 2.6 10.2 8.2 482 15.2 .32 <.02 75th percentile 44 24 150 140 3.6 11.3 8.2 496 19.4 .38 -- -Maximum 66 74 660 210 6.4 11.8 8.4 503 20.1 .43 <.02 

Jacks Fork below 3rd unnamed hollow above Two Rivers (site 185, fig. 7) Number of samples 17 17 17 13 15 17 17 17 17 17 17 17 

Minimum 1 <1 2 7 113 7.6 7.9 280 4.8 .20 <.02 <25th percentile 13 5 18 20 156 9.1 8.2 332 12.8 .26 <.02 Median 30 6 36 44 177 10.5 8.2 340 18.9 .30 <.02 75th percentile 36 11 49 69 233 11.2 8.3 349 21.4 .35 <.02 Maximum 490 640 3,900 1,900 470 13.8 8.6 356 26.4 .63 <.02 

Table 6. Statistical summary of indicator bacteria, physical property, and nutrient data for samples collected on the Jacks Fork, tributaries, and the Eminence November 1999 through December 2000–Continued

Fecal 

coliform 

(col/100 mL)

E. coli

(col/100 mL)

Fecal 

streptococci 

(col/100 mL)

Enterococci 

(col/100 mL)

Discharge

(ft3/s)

Dissolved 

oxygen

(mg/L) pH

Specific 

conductance

(μS/cm)

Water 

temperature

(°C)

Nitrite plus 

nitrate, 

dissolved 

(mg/L as N)

Ammonia, 

dissolved 

(mg/L as N)

Amm

plus o

N, t

(mg/L 
Assessment of Microbiological Contamination 19
20 Assessment of Possible Sources of Microbiological Contamination of the Jacks Fork, Missouri—Phase II

between the distributions of the nine main stem and 

Alley Spring sites was at site 110, which overall tended 

to have larger fecal coliform bacteria densities than the 

other sites. Similar to Phase I of the study, four of the 

five tributaries [not including Alley Spring (site 15)] 

generally had larger fecal coliform densities than the 

main stem sites, but the tributaries again had little effect 

on the fecal coliform bacteria densities in the Jacks 

Fork because of the relatively small discharge of the 

tributaries relative to the discharge of the Jacks Fork 

(Davis and Richards, 2001). The Eminence WWTP 

also was not a contributor to the fecal coliform bacteria 

densities in the Jacks Fork main stem. Out of 13 samples collected from the WWTP, 11 had less than 1 colony of fecal coliform bacteria per 100 mL; the largest 

density was 5 col/100 mL. 

The fecal coliform instantaneous load (the product of bacteria density times stream discharge) is a 

measure of the number of fecal coliform bacteria 

present in the volume of water that passes by a specific 

location in 1 second. Fecal coliform densities and loads 

generally were smallest at sites 5 and 15 (figs. 10, 12), 

which are upstream from the 303(d) reach (fig. 7) and 

are considered to have background concentrations. The 

fecal coliform densities and instantaneous loads generally increased from background concentrations at site 

60 at Eminence, peaked at site 110, and then continued 

to decrease to site 185, which is the downstream-most 

site sampled during Phase II of the study. Large densities and loads at sites 165 and 185 on May 25, 2000, are 

associated with runoff. Generally, the largest densities 

and loads at sites 75 to 150 not related to wet-weather 

flow (figs. 11, 12) were measured August 8 to 10, 2000, 

during a trail ride, when the river discharge was at a 

minimum, water temperatures were at their maximum, 

and river use (trail riding, canoeing, tubing, and swimming) was at a maximum. The most affected site was 

site 110, with large densities and loads that exceeded 

the whole-body-contact standard occurring during trail 

rides that were held May 7 to 13, June 4 to 10, August 

6 to 12, and October 1 to 7, 2000. During the May and 

October trail rides, few canoes and no swimmers or 

tubers were observed using the river. Although trail riders are observed along and in the river from Alley 
Assessment of Microbiological Contamination 21

Spring downstream to site 185, the largest concentration of trail riders occurred between sites 75 and 150, 

with site 110 at an intensely used trail-ride crossing 

outside of the ONSR boundary (fig. 7). 

Swimmers, canoeists, and tubers were observed 

during most sample collection times in June, July, and 

August 2000, including during trail rides. All of the 

sample collection was done Monday through Friday, 

whereas the largest volume of swimmers, canoeists, 

and tubers use the Jacks Fork on the weekend. To investigate the potential effects of large numbers of swimmers, canoeists, and tubers on the fecal coliform 

bacteria densities, a 24-hour sample collection was 

conducted the weekend of July 15 and 16, 2000. Five 

or six samples were collected at sites 5, 15, 28 (a previously unsampled site located about 4 river mi upstream 

from Eminence), 60, 110, and 150 (fig. 7) between Saturday morning until the following Sunday afternoon. 

The samples were analyzed for fecal coliform and E. 

coli bacteria and physical properties, including discharge, DO, pH, specific conductance, and water temperature. In addition, samples were sent to the College 

of Veterinary Medicine at the UMC for ribotyping analysis. No fecal coliform density at any of the sites sampled during the 24-hour period exceeded the wholebody-contact recreation standard (fig. 13). The largest 

measured density of 110 col/100 mL was in a sample 

collected at site 110 at 9:00 a.m. on Sunday. Generally, 

fecal coliform densities were largest in samples collected at site 110; densities ranged from 10 to 110 

col/100 mL, with a median density of 56 col/100 mL in 

six samples. Densities ranged from 6 to 52 col/100 mL 

with a median of 18 col/100 mL at the other five sites. 

The reason for the elevated densities at site 110 may be 

because of the herd of cows that resides in the pasture 

adjoining the river, although the cows were not 

observed crossing the river during sampling. With the 

exception of site 28, which was not sampled between 

12:55 a.m. and 2:25 a.m. or 10:45 a.m. and 11:55 a.m., 

and site 110, fecal coliform densities generally were
22 Assessment of Possible Sources of Microbiological Contamination of the Jacks Fork, Missouri—Phase II
Assessment of Microbiological Contamination 23
24 Assessment of Possible Sources of Microbiological Contamination of the Jacks Fork, Missouri—Phase II
Assessment of Microbiological Contamination 25
26 Assessment of Possible Sources of Microbiological Contamination of the Jacks Fork, Missouri—Phase II
Assessment of Microbiological Contamination 27

largest in the samples collected just after midnight at 

sites 5, 60, and 150. The large number of swimmers, 

canoeists, and tubers using the Jacks Fork on the weekend of July 15 to 16, 2000, did not appear to affect the 

fecal coliform densities.

Because bacteria survive longer in streambed 

sediments than in water, a source of bacteria in the 

water column could be from resuspension of accumulated bacteria from streambed sediments (Marino and 

Gannon, 1991). The fecal bacteria stored in the streambed sediments may be returned to the water column by 

physical disturbance of the streambed sediments by 

processes such as dredging (Grimes, 1980), wind and 

wave action (Lehman and Fogel, 1976), and by swimmers or boaters (Bromel and others, 1978). In the case 

of the Jacks Fork, the mechanical action could be 

caused by the movement of horses through the water or 

by swimmers, canoeists, and tubers. Water and streambed-sediment samples were collected at three sites (5, 

60, and 110) on August 2, 2000, 1 week before a trail 

ride. Additional water and streambed-sediment samples were collected at three sites (110, 150, and 160) on 

August 8, 2000, during a trail ride. These samples were 

analyzed for fecal coliform and E. coli bacteria and 

physical properties, including discharge, DO, pH, specific conductance, and water temperature. In addition, 

samples were sent to the College of Veterinary Medicine at the UMC for ribotyping analysis. The results are 

listed in table 7. Although only site 110 was sampled 

before and during the trail ride, the results indicate that 

the fecal-bacteria densities increased at this site in both 

the streambed sediment and water column during the 

trail ride. Additional water-column and streambed-sediment samples need to be collected to determine if this 

increase routinely occurs.

Analysis of the fecal coliform bacteria data indicates that during the Phase II sampling period, bacteria 

densities not related to wet-weather flow were largest 

during trail rides. During the June 4 to 10 and August 6 

to 12 trail rides, numerous people were camping near 

the river or swimming, canoeing, or tubing in the river, 

which may have contributed to the elevated bacteria 

densities. However, few people were camping or swimming, canoeing, or tubing during the May, October, and 

November trail rides, and bacteria densities were still 

elevated. These elevated densities may be related to 

four factors: (1) physical disturbance of streambed sediments causing resuspension of accumulated bacteria, 

(2) defecation directly into the river, (3) fecal material 

carried into the river on the feet of animals, and (4) 

leakage of sewage effluent from an unknown source 

into the river.
Table 7. Fecal coliform and Escherichia coli densities

in water and streambed-sediment samples collected

at selected sites, August 2000, in colonies per 100 

millilitiers

Site 

number 

(fig. 7)

Fecal coliform Escherichia coli

Water Sediment Water Sediment

August 2, 2000

5 10 88 2 87

60 5 190 1 160

110 11 570 6 340

August 8, 2000

110 310 5,100 148 3,000

150 124 480 23 206

160 1,460 920 1,520 680

28 Assessment of Possible Sources of Microbiological Contamination of the Jacks Fork, Missouri—Phase II

Microbial Source Tracking

Isolates of E. coli were obtained from water samples collected at sites 5 (1 sample), 35 (1 sample), 45 (2 

samples), 60 (5 samples), 80 (2 samples), 110 (5 samples), 150 (3 samples), 160 (1 sample), and 165 (1 sample) and from streambed-sediment samples collected at 

sites 5 (1 sample), 60 (1 sample), 110 (2 samples), 150 (1 

sample), and 160 (1 sample) and were submitted for 

ribotyping analysis (table 8). Whereas most efforts to 

determine the source of bacteria in streams rely on indirect measures of inorganic and organic constituents, 

ribotyping has the promise of directly linking the bacteria 

to their source using DNA ribopatterns. The technique 

relies on the assumption that ribotypes of E. coli from 

various animal species will be unique. However, little is 

known about the temporal and geographic variability of 

ribotypes within a single animal group or the potential 

sharing of ribotypes between various animals. Hartel and 

others (2002) determined that ribotypes for cows in Idaho 

and Georgia differed significantly. Sargeant and others 

(1999) showed that wild deer foraging in fields where 

dairy cows were pastured became colonized with the 

identical strain of E. coli carried by the cows. 

Ribotyping analysis involves the use of multivariate statistical methods to compare patterns in large 

data sets. The method compares the degree of similarity 

of the patterns from unknown samples to known patterns in a database. For this study, the unknowns were 

compared to known patterns obtained from sewage, 

horses, and cows (table 8). Patterns from sewage differ 

from patterns obtained directly from human fecal material. Sources of sewage in the study area could include 

the Eminence WWTP or campground pit-toilet or septic-system effluent. The three choices were based on 

the presence of all of these sources in the study area. 

Matching of patterns was made using discriminate 

analysis, and a pattern was considered to be a match if 

the probability of a match was 70 percent or higher. If 

no pattern was matched, the isolate was considered to 

come from unknown sources, which could include 

domesticated animals such as pigs, dogs, cats, or chickens, or wild animals such as deer, raccoons, geese, and 

ducks. As additional patterns are added to the database 

of “known” patterns, the degree of similarity between 

unknown and known patterns constantly changes. 

Because of the large degree of uncertainty in the 

method, results of the method should be treated as 

experimental for the purpose of this study, and interpretations of these data were made in conjunction with 

other data and information.

The samples for ribotyping analysis were collected in 2000 on April 6 at the beginning of the recreational season; June 8 during the June 4 to 10 trail ride; 

July 16 during intense swimming, canoeing, and tubing 

use; August 2, 1 week before a trail ride; and August 8 

during the August 6 to 12 trail ride. Ribopatterns were 

obtained from 65 E. coli isolates from 13 water samples 

and from 23 E. coli isolates from 4 streambed-sediment 

samples (table 8). No isolates were obtained from 

water and/or streambed-sediment samples collected at 

site 110 on June 8; sites 60, 110, and 150 on July 16; 

sites 5, 60, and 110 on August 2; and at sites 60 and 160 

on August 8 because of a failed E. coli biochemical 

confirmation test, no E. coli colonies were isolated for 

ribotyping, or the DNA fragments from the E. coli isolates smeared during gel electrophoresis.

The isolates were compared to several hundred 

isolates obtained from animal sources at the UMC 

School of Veterinary Medicine farms, selected locations in southwest Missouri, and the Eminence area. 

Results indicated that of the 65 E. coli isolates obtained 

from the water samples, 26 (40 percent) were identified 

as originating from sewage, 19 (29 percent) from horse, 

7 (11 percent) from cow, and 13 (20 percent) from 

unknown sources. Of the 23 isolates from the streambed-sediment samples, 9 (39 percent) were identified 

as originating from sewage, 8 (35 percent) from horse, 

3 (13 percent) from cow, and 3 (13 percent) from
Microbial Source Tracking 29 a Samples collected and processed by the U.S. Geological Survey. b Samples collected by the U.S. Geological Survey and processed by the College of Veterinary Medicine at the University of Missouri in Columbia, Missouri. c Ribotyping analysis failed for technical reasons (smeared pattern). d No Escherichia coli colonies isolated for ribotyping. e Failed biochemical confirmation test.

Table 8. Number of Escherichia coli isolates in water or streambed-sediment samples assigned to various sources by ribotyping[col/100 mL, colonies per 100 milliliters of sample; E. coli, Escherichia coli; W, water; <, less than; --, no data available; S, streambed sediment]

Site

(fig. 7) Date Sample matrix

Fecal coliform 

(col/100 mL)a

E. coli 

(col/100 mL)a

Total number of 

isolatesb

Possible animal source using three kno

(probability of 0.70 or larger)

Sewage Horse Cow 

45 04/06/00 W 27 20 2 0 0 1 60 04/06/00 W 27 15 1 0 0 1 

110 04/06/00 W <1 <1 3 0 1 1 

45 06/08/00 W 380 310 6 0 5 0 60 06/08/00 W 28 27 10 7 2 0 80 06/08/00 W 56 43 10 5 1 1 

110 06/08/00 W 240 80 c0 -- -- -- 

150 06/08/00 W 46 3 2 2 0 0 

60 07/16/00 W 14 <2 d0 -- -- -- 

110 07/16/00 W 10 12 d0 -- -- -- 

150 07/16/00 W 8 2 d0 -- -- -- 

5 08/02/00 W 10 2 e0 -- -- -- 

08/02/00 S 88 87 6 2 3 0 

60 08/02/00 W 5 1 d0 -- -- -- 

08/02/00 S 190 160 e0 -- -- -- 

110 08/02/00 W 11 6 d0 -- -- -- 

08/02/00 S 570 340 3 2 1 0 

35 08/08/00 W 180 48 6 1 3 1 60 08/08/00 W 25 13 c0 -- -- -- 

80 08/08/00 W 210 180 4 4 0 0 

110 08/08/00 W 310 148 5 4 1 0 

08/08/00 S 5,100 3,000 9 5 3 1 

150 08/08/00 W 124 23 8 1 5 1 

08/08/00 S 480 206 5 0 1 2 

160 08/08/00 W 1,460 1,520 7 2 0 1 

08/08/00 S 920 680 d0 -- -- -- 

165 08/08/00 W 96 20 1 0 1 0 
30 Assessment of Possible Sources of Microbiological Contamination of the Jacks Fork, Missouri—Phase II

unknown sources. The overall results indicate that the 

predominant sources of E. coli bacteria were sewage 

and horses.

At the beginning of the recreational season, no 

matches were classified as human. Of the six isolates 

obtained from three samples collected on April 6, three 

were classified as cow, one as horse, and two unknown 

(fig. 14). The small number of isolates is consistent 

with the small fecal coliform and E. coli densities 

observed in these samples at these sites and can be considered representative of base-flow conditions during 

the early part of the recreational season (table 8, fig. 8). 

Twenty-eight isolates were obtained from samples collected June 8 during a trail ride. Fourteen isolates were assigned to sewage, 8 to horse, 1 to cow, and 

5 to unknown sources (fig. 14). Although some trail 

riders do ride along the Jacks Fork upstream from site 

75, most trail-ride activity occurs between sites 75 and 

165. On June 8, no isolates were obtained at site 110, 

and only two were obtained at site 150. Site 45 is 

located on Storys Creek, a small (5.34 mi2

), predominantly agricultural basin. Ribotyping results for site 45 

indicate that five of the six isolates were assigned to 

horse. Cows have been observed grazing in pastures 

located immediately upstream from the site.

In the samples collected August 2, 1 week before 

a trail ride, isolates were obtained only from streambed-sediment samples collected at sites 5 and 110. Of 

the nine isolates, four were assigned to sewage, four to 

horse, and one to an unknown source (fig. 14). The 

small number of isolates is consistent with the small 

fecal coliform and E. coli densities in these samples at 

these sites and can be considered representative of 

base-flow conditions during the recreational season 

(table 8, fig. 8). 

The most complete set of results was obtained 

from the samples collected August 8 during a trail ride. 

Of the 31 isolates obtained from 7 water samples, 12 

were classified as originating from sewage, 10 from 

horse, 3 from cow, and 6 from unknown sources. Of the 

14 isolates obtained from streambed-sediment samples, 5 were classified as originating from sewage, 4 

from horse, 3 from cow, and 2 from unknown sources 

(fig. 14). At the sites most affected by trail-ride activity 

(sites 80, 110, 150, and 160), water sample isolates 

identified as originating from sewage accounted for 46 

percent of the total number of isolates, and isolates 

identified as originating from horse accounted for 25 

percent of the total. Of the 14 isolates obtained from 

streambed-sediment samples collected at sites 110 and 

150, 36 percent were identified as originating from 

sewage, and 29 percent were identified as originating 

from horse.

As discussed previously, ribotyping results 

should be treated as experimental for the purpose of 

this study because of the large degree of uncertainty in 

the method. However, based on the results of comparing unknown patterns to patterns for sewage, horses, 

and cows, sewage and horses appear to be possible 

sources of E. coli bacteria in the Jacks Fork. Additional 

MST using ribotyping and other techniques needs to be 

done to further pinpoint E. coli sources. Since 2000, 

when these samples were analyzed, MST technology, 

including the software available to analyze the results, 

has improved. Obtaining additional samples of known 

sources from the Eminence area also would assist in 

MST efforts.

WATER-QUALITY CHARACTERISTICS

The measurement of physical properties and 

nutrient concentrations in surface water can provide 

additional information on the general condition of the 

stream and the effects of hydrologic events and recreational uses on stream water quality. The physical characteristics and chemical composition of the stream can 

indicate whether the stream environment can support 

diverse aquatic life and is desirable for recreational use.

Physical Properties

The physical properties measured at each site 

included DO, pH, specific conductance, and temperature. A statistical summary of the physical-property 

data is given in table 6. 

Dissolved Oxygen

The distribution of DO concentrations on the 

Jacks Fork and tributaries and Eminence WWTP is 

shown in figure 15. Median DO concentrations at the 

nine main stem sites and Alley Spring ranged from 9.9 

to 10.7 mg/L (table 6) with no statistically significant 

differences in the distributions. No DO concentration 

on the Jacks Fork main stem was less than the minimum of 5 mg/L required by the State of Missouri to 

sustain a cool-water fishery (Missouri Department of 

Natural Resources, 2000). Although the DO concentra-
Water-Quality Characteristics 31
32 Assessment of Possible Sources of Microbiological Contamination of the Jacks Fork, Missouri—Phase II

tion was extremely small (minimum of 1.1 mg/L) in the 

effluent from the Eminence WWTP, no DO effects 

were observed at site 75 immediately downstream from 

the WWTP, probably because of the small volume of 

the effluent. Variations in DO concentrations appear to 

be seasonal, with the largest DO concentrations tending 

to occur in the cooler months, or related to the time of 

day the DO concentration was determined. 

pH

The median pH for Jacks Fork main stem sites 

ranged from 8.1 to 8.3 (fig. 16, table 6) with no statistically significant differences between the sites. The 

median pH for samples collected at Alley Spring was 

7.7, which was significantly different (p-value less than 

0.05) than the values of the main stem sites. This is not 

unexpected because carbon dioxide in ground water 

normally occurs at a much higher partial pressure than 

in the earth’s atmosphere causing the pH in ground 

water to be lower than the concentration in streams. 

When ground water is exposed to the atmosphere, carbon dioxide will escape and the pH will increase 

(Freeze and Cherry, 1979, p. 140). The median pH of 

the samples collected at the Eminence WWTP was 7.7, 

and although the median pH increased steadily at sites 

downstream from the WWTP (median of 8.1, 8.2, and 

8.3 at sites 75, 80, and 110), this increase probably is 

not directly related to the effluent. The median pH at 

site 60 located immediately upstream from site 70 also 

was 8.1. Variations in pH primarily appear to be related 

to hydrologic events or related to the time of day the pH 

measurement was taken because the smallest pH values 

tended to occur in samples collected at higher flows or 

in the morning.
Water-Quality Characteristics 33

Specific Conductance

Median specific conductance values ranged from 

327 to 340 μS/cm [microsiemens per centimeter at 25 

°C (degrees Celsius)] (fig. 17, table 6) at the Jacks Fork 

main stem sites with no statistically significant differences between the sites. The median specific conductance value at Alley Spring (site 15; 310 μS/cm) was 

significantly smaller (p-value less than 0.05) than the 

values of the main stem sites, but did not appear to have 

a dilution effect on the Jacks Fork. Likewise, the 

median specific conductance at the Eminence WWTP 

(site 70; 752 μS/cm) does not cause a significant 

increase in the specific conductance in the Jacks Fork 

immediately downstream from the WWTP at site 75. 

Variations in specific conductance values (figs. 8 and 

18, table 3) appear to be related primarily to hydrologic 

events, with the smallest specific conductance values 

occurring at the largest flows.

Water Temperature

Median water temperature for the Jacks Fork 

main stem sites ranged from 17.9 to 21.9 °C (table 6) 

with no significant differences between the sites. The 

median water temperature for Alley Spring (site 15) 

was 14.6 °C (typical of spring water), which was significantly smaller (p-value less than 0.05) than the values 

of the main stem sites. Variation in water temperature 

throughout the sampling period was because of seasonal changes, with the lowest temperatures occurring 

in the winter and the highest temperatures occurring in 

the summer (fig. 19). The water temperature of Alley 

Spring did not vary with season. 

Nutrients

The fixation of atmospheric nitrogen by plants 

and animals, atmospheric deposition of nitrogen, the 

dissolution of phosphorus-bearing rocks or minerals in 
34 Assessment of Possible Sources of Microbiological Contamination of the Jacks Fork, Missouri—Phase II

the soils, and organic matter, including soil organic 

matter and decaying plants and animals, are natural 

sources for nitrogen and phosphorus in streams. 

Anthropogenic sources include sewage discharges, fertilizers, animal waste, and septic tanks (Hem, 1985). 

Background concentrations of nitrogen and phosphorus in streams generally are small because the dissolved 

forms of the two elements are assimilated rapidly by 

plants and bacteria. Excessive concentrations of nutrients can have detrimental effects on desired uses of 

water and may indicate possible contamination. 

Anthropogenic sources of nutrients in the study area 

are the Eminence WWTP, septic systems, cows, and 

horses. 

The nutrients analyzed included dissolved species of nitrite plus nitrate, ammonia, and orthophosphate and total species of ammonia plus organic 

nitrogen and phosphorus. Summary statistics of the 

nutrient data for the Jacks Fork are listed in table 6. The 

dissolved nitrite plus nitrate and total phosphorus data 

will be discussed in detail in the following sections. 

Differences in dissolved ammonia and orthophosphate 

and total ammonia plus organic nitrogen were not statistically significant between sites.

Dissolved Nitrite Plus Nitrate

On the main stem of the Jacks Fork, dissolved 

nitrite plus nitrate concentrations were statistically 

smaller (p-value less than 0.05) at the most-upstream 

site above Alley Spring (site 5; median 0.11 mg/L) and 

increased below Alley Spring at Eminence (site 60; 

median 0.30 mg/L) (fig. 20, table 6). This increase also 

was observed during Phase I data collection (Hauck 

and Nagel, 2000) and is caused by high dissolved 

nitrite plus nitrate concentrations in Alley Spring (site 

15; median 0.58 mg/L), whose input at lower main 

stem flows will double the discharge of the Jacks Fork. 

A small, but not statistically significant, increase in dissolved nitrite plus nitrate concentrations occurred at 
Wastewater Organic Compounds 35

site 75 (median 0.38 mg/L) immediately downstream 

from the WWTP (site 70; median 16.2 mg/L). The 

effect diminishes quickly downstream from the WWTP 

at sites 80 (median 0.34 mg/L), 110 (median 0.31 

mg/L), 140 (median 0.28 mg/L), and 150 (median 0.28 

mg/L). The median concentration of dissolved nitrite 

plus nitrate was 0.30 mg/L at sites 165 and 185. The 

minimal effects of the Eminence WWTP also were 

observed in Phase I (Hauck and Nagel, 2000). Unlike 

fecal coliform bacteria, dissolved nitrite plus nitrate 

concentrations appear to be related mostly to hydrologic conditions and do not increase during recreational 

activities (fig. 21). 

Total Phosphorus

Similar results were observed with total phosphorus as with dissolved nitrite plus nitrate. On the 

main stem of the Jacks Fork, total phosphorus concentrations again tended to be smallest at the most 

upstream site above Alley Spring (site 5; median less 

than 0.004 mg/L), but did not increase significantly 

downstream from Alley Spring (site 15; median 0.009 

mg/L) at site 60 (median less than 0.004 mg/L) (fig. 22, 

table 6). Similar results also were reported during 

Phase I data collection (Hauck and Nagel, 2000). A statistically significant increase in total phosphorus concentrations occurred at site 75 (median 0.009 mg/L) 

immediately downstream from the WWTP (site 70; 

median 3.0 mg/L). Similar to dissolved nitrite plus 

nitrate concentrations, the effect diminishes quickly 

downstream from the WWTP at sites 80 (median 0.005 

mg/L), 110 (median 0.005 mg/L), 140 (median 0.004 

mg/L), and 150 (median 0.004 mg/L). Median concentrations of total phosphorus were 0.005 and 0.004 mg/L 

at sites 165 and 185. The effects of the Eminence 

WWTP also were observed in Phase I (Hauck and 

Nagel, 2000).

Wastewater Organic Compounds

Trace quantities of organic compounds commonly associated with municipal or domestic sewage 

effluent were detected in samples collected at sites 5, 

15, 60, 80, and 110 (tables 5, 9). Most of the detected 

concentrations were estimated because they were less 

than the MRL. Octylphenol monoethoxylate (OPEO1), 

nonylphenol monoethoxylate (NPEO1), and bispenol 

A were detected in the associated laboratory blanks. In 

the cases of OPEO1 and bisphenol A, the laboratoryblank concentrations were larger than the concentration 
36 Assessment of Possible Sources of Microbiological Contamination of the Jacks Fork, Missouri—Phase II

detected in the environmental sample, thereby making 

data interpretation impossible. Some of the compounds 

detected have sources other than sewage effluent. Polynuclear aromatic hydrocarbons (PAHs) can be associated with asphalt roads, water proofing of concrete 

foundations, asphalt roof shingles, and fuel combustion. Plasticizers are extremely common in an industrialized society because they are in many products and, 

therefore, are commonly reported laboratory contaminants. N,N-diethyltoluamide, or DEET, is a common 

ingredient in insect repellents, and its presence in the 

Jacks Fork would not be unexpected.

Of the organic compounds detected in the water 

samples from the Jacks Fork and Alley Spring, the best 

indicators of municipal or domestic sewage effluent are 

the non-ionic detergent metabolites (NPEO1, OPEO1, 

and para-nonylphenol), the disinfectant phenol, and 

caffeine. However, possible sources of these compounds could be the numerous campers, swimmers, 

and canoeists that were present in the area on July 16 

and August 10, 2000, when the wastewater organic 

compound samples were collected. Associated fecal 

coliform bacteria densities were relatively small in the 

samples collected on July 16 and August 10, or both, at 

sites 5 (10 col/100 mL), 15 (22 col/100 mL), 60 (14 and 

25 col/100 mL), and 110 (10 col/100 mL), indicating a 

source other than sewage. The fecal coliform density at 

site 80 was 210 col/100 mL, but the only compounds 

detected were caffeine [estimated 0.08 μg/L (microgram per liter)] and N,N-diethyltoluamide (estimated 

0.038 μg/L). At the time of sample collection, numerous swimmers and tubers were in the water, which may 

account for the presence of these compounds.

SUMMARY

In 1998, an 8-mile reach of the Jacks Fork was 

included on Missouri’s list of impaired waters as 

required by Section 303(d) of the Federal Clean Water 

Act. The identified pollutant on the Jacks Fork was 

fecal coliform bacteria, whose presence in large numbers indicates contamination by fecal wastes of humans 

and other warm-blooded animals. The standard for safe 

whole-body-contact recreation is 200 col/100 mL (colonies per 100 milliliters) of sample. Potential sources 

of fecal contamination to the Jacks Fork include a 

wastewater treatment plant; campground pit-toilet or 

septic-system effluent; a large commercial, cross-country horseback trail riding facility; canoeists, boaters, 

and tubers; and cows. The U.S. Geological Survey, in 

cooperation with the National Park Service, conducted 
Summary 37

a study to better understand the extent and sources of 

microbiological contamination within the Jacks Fork 

from Alley Spring to the mouth, which includes the 8-

mile 303(d) reach. Identification of the sources would 

provide the National Park Service and the State of Missouri with the information needed to craft a solution of 

abatement, regulation, prevention, and mitigation with 

the end result being the removal of the Jacks Fork from 

the 303(d) list. 

During Phase II of the study, 15 sites were sampled approximately monthly from November 1999 

through April 2000 and from September 2000 through 

December 2000 and twice per month from May 2000 

through August 2000. An additional site was sampled 

one time. Samples were collected mostly during baseflow conditions during a variety of nonrecreational and 

recreational season river uses, including canoeing, 

swimming, tubing, and horseback riding. Samples 

were analyzed for fecal indicator bacteria [including 

fecal coliform, Escherichia coli (E. coli), fecal streptococci, and enterococci], physical properties, nutrients, 

and wastewater organic compounds. Additional samples were collected and sent to the College of Veterinary Medicine at the University of Missouri in 

Columbia, Missouri, for ribotyping analysis.

During the sampling period, the whole-bodycontact recreation standard was exceeded from 

upstream to downstream at sites 80, 110, and 140 on 

August 10, 2000 (210, 340, and 210 col/100 mL), and 

also at site 110 on May 11, June 7, and October 3, 2000, 

(340, 240, and 780 col/100 mL). Each of these 

exceedences occurred in samples collected during trail 

rides.

The only statistically significant difference 

between the overall fecal coliform bacteria density distributions of the Jacks Fork main stem sites was at site 

110, which tended to have larger fecal coliform bacteria densities than the other main stem sites. Four of the 

five tributaries (not including Alley Spring) sampled 

generally had larger fecal coliform densities than the 
38 Assessment of Possible Sources of Microbiological Contamination of the Jacks Fork, Missouri—Phase II

main stem sites, but the tributaries had little effect on 

the fecal coliform bacteria densities in the Jacks Fork 

because of the relatively small discharge of the tributaries relative to the discharge of the Jacks Fork. The Eminence Wastewater Treatment Plant also was not a 

contributor to the fecal coliform bacteria densities in 

the Jacks Fork.

The fecal coliform densities and instantaneous 

loads generally increased from background concentrations at site 60 at Eminence, peaked at site 110, and 

then continued to decrease to site 185, which is the 

downstream-most site sampled. The largest concentration of trail riders occurred between sites 75 and 150, 

and site 110 is located at an intensely used trail-ride 

crossing outside of the Ozark National Scenic Riverways boundary. Generally, the largest densities and 

loads at sites 75 to 150 not related to wet-weather flow 

were observed during a trail ride held August 6 to 12, 

2000. 

To investigate the effects that large numbers of 

swimmers, canoeists, and tubers would have on the 

fecal coliform densities, a 24-hour sample collection 

was conducted the weekend of July 15 to 16, 2000. 

Between five or six samples were collected at sites 5, 

15, 28, 60, 110, and 150 between Saturday morning 

until the following Sunday afternoon. No fecal 

coliform density at any of the sites sampled during the 

24-hour period exceeded the whole-body-contact recreation standard. The largest observed density of 110 

col/100 mL was in a sample collected at site 110 at 9:00 

a.m. on Sunday. The large number of swimmers, 

canoeists, and tubers using the Jacks Fork did not 

appear to affect the fecal coliform densities.

Because bacteria survive longer in streambed 

sediments than in water, a source of bacteria in the 

water column could be from resuspension of accumulated bacteria from streambed sediments. Water and 

streambed-sediment samples were collected at three 

sites (5, 60, and 110) on August 3, 2000, one week 

before a trail ride. Additional water and streambed-sediment samples were collected at three sites (110, 150, 

and 160) during a trail ride. Results indicate that fecalbacteria densities increased substantially in the streambed sediment and water column during the trail ride.

Analysis of the fecal coliform bacteria data indicates that during the Phase II sampling period bacteria 

densities not related to wet-weather flow were largest 

during trail rides. During the June 4 to 10 and August 6 

to 12 trail rides, numerous people were camping near 

the river or swimming, canoeing, or tubing in the river, 

which may have contributed to the elevated bacteria 

densities. However, few people were camping or swim-
Summary 39

ming, canoeing, or tubing during the May, October, and 

November trail rides, and bacteria densities were still 

elevated. The reasons for these elevated densities may 

be related to four factors: (1) physical disturbance of 

streambed sediments causing resuspension of accumulated bacteria, (2) defecation directly into the river, (3) 

fecal material carried into the river on the feet of animals, and (4) leakage of sewage effluent from an 

unknown source into the river. 

Sixty-five E. coli isolates obtained from water 

samples collected at 9 sites (5, 35, 45, 60, 80, 110, 150, 

160, and 165) and 23 E. coli isolates obtained from 

streambed-sediment samples collected at 5 sites (5, 60, 

110, 150, and 160) were submitted for ribotyping analysis. Samples were collected in 2000 during a variety 

of nonrecreational and recreational season river uses, 

including trail rides, canoeing, tubing, and swimming. 

Ribotypes were compared to a library of ribotypes from 

three known sources: sewage, horses, and cows. Of the 

65 isolates from water samples, 40 percent were identified as originating from sewage, 29 percent from 

horse, 11 percent from cow, and 20 percent from 

unknown sources. Of the 23 isolates from streambedsediment samples, 39 percent were identified as originating from sewage, 35 percent from horse, 13 percent 

from cow, and 13 percent from unknown sources. 

Ribotyping results should be treated as experimental 

for the purpose of this study because of the large degree 

of uncertainty in the method. However, based on the 

results of comparing unknown patterns to patterns for 

sewage, horses, and cows, sewage and horses do appear 

to be possible sources of E. coli in the Jacks Fork.

An analysis of the physical property data, including dissolved oxygen, pH, specific conductance, and 

temperature, for the main stem of the Jacks Fork and 

Alley Spring indicated that no statistically significant 

differences occurred among the main stem sites for any 

of the physical properties. The median pH, specific
40 Assessment of Possible Sources of Microbiological Contamination of the Jacks Fork, Missouri—Phase II Table 9. Selected wastewater organic compound concentrations and fecal coliform densities in water samples collected on July 16 

[All concentrations in micrograms per liter; bold numbers indicate that compound was detected; NPEO1, nonylphenol monoethoxylate; OPEO1, octylphenol monoper 100 milliliters; <, less than; E, estimated; b, detected in laboratory blank] Date NPEO1 OPEO1 para- Nonylphenol Phenol N,N- diethyltoluamide Caffeine Bisphenol A Ethanol, 2-butoxy-, phosphate Naphtha

Jacks Fork above Alley Spring (site 5, fig. 7) 07/16/00 <1.00 <0.10 <0.50 E0.44 <0.040 <0.08 <0.09 <0.20 <0.02 

Alley Spring at Alley (site 15, fig. 7) 07/16/00 <1.00 <.10 <.50 E.45 <.040 <.08 <.09 <.20 <.02 

Jacks Fork at Eminence (site 60, fig. 7) 07/16/00 <1.00 <.10 E.95 E.34 E.018 <.08 <.09 <.20 <.02 

08/10/00 <1.00 <.10 <.50 E.20 E.034 <.08 .45 b .21 .02 

Jacks Fork above 2nd unnamed hollow below Eminence (site 80, fig. 7) 08/10/00 <1.00 <.10 <.50 <.25 E.038 E.08 .30 b <.20 <.02 

Jacks Fork above Lick Log Hollow below Eminence (site 110, fig. 7) 07/16/00 E1.26 b E.06 b E.95 E1.03 .044 <.08 <.09 <.20 <.02 
References 41

conductance, and temperature for samples collected at 

Alley Spring were significantly smaller than the values 

for the main stem sites. In samples collected from the 

Eminence Wastewater Treatment Plant, the median dissolved oxygen concentration and pH were smaller than 

the medians for the main stem sites, and the median 

specific conductance was larger than the medians for 

the main stem sites. However, no effects from the Eminence Wastewater Treatment Plant were observed 

immediately downstream at site 75, probably because 

of the small volume of the effluent. Unlike fecal 

coliform bacteria, most variation in dissolved oxygen, 

pH, specific conductance, and temperature values was 

related to seasonal changes, time of day the samples 

were collected, or hydrologic conditions and not to recreational activities.

On the main stem of the Jacks Fork, dissolved 

nitrite plus nitrate concentrations were statistically 

smaller at the most upstream site above Alley Spring 

[site 5; median 0.11 mg/L (milligrams per liter)] and 

increased below Alley Spring at Eminence (site 60; 

median 0.30 mg/L). A small, but not statistically significant, increase in dissolved nitrite plus nitrate concentrations occurred at site 75 (median 0.38 mg/L) 

immediately downstream from the Eminence Wastewater Treatment Plant (site 70; median 16.2 mg/L). The 

effect diminished quickly downstream from the plant. 

Unlike fecal coliform bacteria, dissolved nitrite plus 

nitrate concentrations were related mostly to hydrologic conditions and did not increase during recreational activities. Similar results were observed with 

total phosphorus. However, the increase in total phosphorus concentrations at site 75 immediately downstream from the Eminence Wastewater Treatment Plant 

was statistically significant, but the effect diminished 

quickly downstream.

Trace quantities of organic compounds commonly associated with municipal or domestic sewage 

effluent were detected in samples collected at sites 5, 

15, 60, 80, and 110. Octylphenol monoethoxylate and 

bisphenol A were detected in associated laboratory 

blanks at concentrations larger than the concentration 

detected in the environmental sample, thereby making 

interpretation of the data impossible. Other detected 

compounds, such as polynuclear aromatic hydrocarbons, have sources other than sewage effluent. The best 

indicators of municipal or domestic sewage effluent 

were the non-ionic detergent metabolites (nonylphenol 

monoethoxylate, octylphenol monoethoxylate, and 

para-nonylphenol), phenol, and caffeine. However, 

possible sources of these compounds, which were 

detected in one or more of the samples, could be the 

numerous campers, swimmers, and canoeists that were 

present when the samples were collected.

Although both Phase I and II sampling indicate 

that fecal coliform densities tend to increase to sometimes unacceptable levels during trail rides, the exact 

causes for this increase and the sources of the fecal 

coliform bacteria have not been positively identified. 

Phase III sampling has continued at 10 sites plus the 

Eminence Wastewater Treatment Plant since January 

2001, with specific recreational activities (canoeing 

and trail rides) being the primary focus of the sampling 

efforts. Additional work is being done to examine the 

role that storage of fecal coliform in the streambed sediments and then subsequent resuspension may contribute to the overall concern and to try to positively 

identify the sources of the bacteria using ribotyping and 

other Microbial Source Tracking techniques. 

REFERENCES

Aley, C., and Aley, T.J., 1987, Ground water study, Ozark 

National Scenic Riverways: Ozark Underground Laboratory, Protem, Missouri, v. I, 227 p. 

Barks, J.H., 1978, Water quality in the Ozark National Scenic Riverways, Missouri: U.S. Geological Survey 

Water-Supply Paper 2048, 57 p.

Bromel, M., Saylor, G., Coles, G., Zimmerman, J., and 

Johnson, K., 1978, Bacteriological analyses of Lake 

Metigoshe water and sediments: Fargo, North Dakota 

State University, Agricultural Experiment Stations Bulletin 507, 14 p.

Carson, C.A., Shear, B.L., Ellersieck, M.R., and Asfaw, A., 

2001, Identification of fecal Escherichia coli from 

humans and animals by ribotyping: Applied and Environmental Microbiology, v. 67, no. 4, p. 1,503–1,507.

Davis, J.V., and Bell, R.W., 1998, Water-quality assessment 

of the Ozark Plateaus study unit, Arkansas, Kansas, 

Missouri, and Oklahoma—Nutrients, bacteria, organic 

carbon, and suspended sediment in surface water, 

1993–95: U.S. Geological Survey Water-Resources 

Investigations Report 98–4164, 95 p.

Davis, J.V., and Richards, J.M., 2001, Assessment of microbiological contamination of the Jacks Fork within the 

Ozark National Scenic Riverways, Missouri—Phase I: 

U.S. Geological Survey Fact Sheet 026–01, 6 p.

Doyle, J.D., Runnicliff, B., Kramer, R., Kuehl, R., and Brickler, S.K., 1992, Instability of fecal coliform populations 

in waters and bottom sediments at recreational beaches 

in Arizona: Water Research, v. 26, p. 979–988.
42 Assessment of Possible Sources of Microbiological Contamination of the Jacks Fork, Missouri—Phase II

Dufour, A.P., 1977, Escherichia coli—The fecal coliform, in 

Hoadley, A., and Dutka, B.J., eds., Bacterial indicators/health hazards associated with water, 1977: American Society for Testing and Materials, ASTM STP 635, 

p. 48–58.

Dugan, J.T., and Peckenpaugh, J.M., 1985, Effects of climate, vegetation, and soils on consumptive water use 

and ground-water recharge to the Central Midwest 

regional aquifer system, Mid-Continent United States: 

U.S. Geological Survey Water-Resources Investigations Report 85–4236, 77 p.

Edwards, T.K., and Glysson, G.D., 1998, Field methods for 

measurement of fluvial sediment: U.S. Geological Survey Techniques of Water-Resources Investigations, 

book 3, chap. C2, 80 p.

Emrie, G.E., 1986, Fecal coliform as an indicator of water 

quality and recreational carrying capacity at Ozark 

National Scenic Riverways: Springfield, Southwest 

Missouri State University, unpublished M.S. thesis, 

65 p.

Fenneman, N.M., 1938, Physiography of eastern United 

States: New York, McGraw-Hill Book Co., Inc., 689 p.

Fishman, M.J., ed., 1993, Methods of analysis by the U.S. 

Geological Survey National Water Quality Laboratory—Determination of inorganic and organic constituents in water and fluvial sediments: U.S. Geological 

Survey Open-File Report 93–125, 217 p.

Fishman, M.J., and Friedman, L.C., eds., 1989, Methods for 

determination of inorganic substances in water and fluvial sediments: U.S. Geological Survey Techniques of 

Water-Resources Investigations, book 5, chap. A1, 

545 p.

Francy, D.S., and Darner, R.A., 1998, Factors affecting 

Escherichia coli concentrations at Lake Erie public 

bathing beaches: U.S. Geological Survey WaterResources Investigations Report 98–4241, 41 p.

Freeze, R.A., and Cherry, J.A., 1979, Groundwater: Englewood Cliffs, New Jersey, Prentice-Hall, Inc., 604 p.

Grimes, D.J., 1980, Bacteriological water quality effects of 

hydraulically dredging contaminated upper Mississippi 

River bottom sediment: Applied and Environmental 

Microbiology, v. 39, no. 4, p. 782–789.

Hartel, P.G., Segars, W.I., Stern, N.J., Steiner, J., and 

Buchan, A., 1999, Ribotyping to determine the host origin of Escherichia coli isolates in different water samples, in Olsen, D.S., and Potyondy, J.P., eds., Wildland 

hydrology, 1999: American Water Resources Association, TPS-99-3, p. 377–382.

Hartel, P.G., Summer, J.D., Hill, J.L., Collins, J.V., Entry, 

J.A., and Segars, W.I., 2002, Biogeographic variability 

of Escherichia coli ribotypes from Idaho and Georgia: 

Journal of Environmental Quality, v. 31, no. 4, p. 1,273–

1,278. 

Hauck, H.S., and Nagel, C.D., 2000, Water resources data 

Missouri water year 1999: U.S. Geological Survey 

Water-Data Report MO–99–1, 390 p.

——— 2001, Water resources data Missouri water year 

2000: U.S. Geological Survey Water-Data Report MO–

00–1, 430 p.

Helsel, D.R., and Hirsch, R.M., 1992, Statistical methods in 

water resources: Amsterdam, Netherlands, Elsevier, 

522 p.

Hem, J.D., 1985, Study and interpretation of the chemical 

characteristics of natural water: U.S. Geological Survey 

Water-Supply Paper 2254, 263 p.

Lehman, G.S., and Fogel, M.M., 1976, Investigation of bacteriological pollution of recreational waters in Arizona: 

Tucson, University of Arizona Department of Watershed Management and University of Arizona School of 

Renewable Natural Resources.

Marino, R.P., and Gannon, J.J., 1991, Survival of fecal 

coliforms and fecal streptococci in storm drain sediments: Water Research, v. 25, no. 9, p. 1,089–1,098.

Missouri Department of Natural Resources, 1998, Water 

quality investigation of the Jacks Fork River, Shannon 

and Texas Counties, May–November 1998: Jefferson 

City, Missouri, Division of Environmental Quality, 

Environmental Services Program, 133 p.

——— 2000, Missouri water quality standards—Chapter 7, 

Water quality: Jefferson City, Missouri, Clean Water 

Commission, 136 p.

Missouri Spatial Data Information Service, 2001, Land 

use/land cover data: Columbia, Missouri, accessed July 

2001, at URL http://msdis.missouri.edu/pub/lulc.

Myers, D.N., Koltun, G.F., and Francy, D.S., 1998, Effects of 

hydrologic, biological, and environmental processes on 

sources and concentrations of fecal bacteria in the 

Cuyahoga River, with implications for management of 

recreational waters in Summit and Cuyahoga Counties, 

Ohio: U.S. Geological Survey Water-Resources Investigations Report 98–4089, 45 p.

Myers, D.N., and Wilde, F.D., eds., 1997, National field 

manual for the collection of water-quality data—Biological indicators: U.S. Geological Survey Techniques 

of Water-Resources Investigations, book 9, chap. A7, 

38 p.

National Oceanic and Atmospheric Administration, 1999, 

Climatologic data annual summary, Missouri: 

Asheville, North Carolina, National Climatic Data 

Center.

——— 2000, Climatologic data annual summary, Missouri: 

Asheville, North Carolina, National Climatic Data 

Center.

Parveen, S., Portier, K.M., Robinson, K., Edmiston, L., and 

Tamplin, M.L., 1999, Discriminant analysis of ribotype 

profiles of Escherichia coli for differentiating human 
References 43

and nonhuman sources of fecal pollution: Applied and 

Environmental Microbiology, v. 65, no. 7, p. 3,142–

3,147.

Pommequy, M., Guillard, J.F., Dupray, E., Derrien, A., Le 

Guyader, F., and Cormier, M., 1992, Enteric bacteria 

survival factors: Water Science and Technology, v. 25, 

p. 93–103.

Roszak, D.B., and Colwell, R.R., 1987, Survival strategies of 

bacteria in the natural environment: Microbiological 

Reviews, v. 51, p. 365–379.

Sargeant, J.M., Hafer, D.J., Gillispie, J.R., Oberst, R.D., and 

Flood, S.J., 1999, Prevalence of Escherichia coli

O157:H7 in white-tailed deer sharing rangeland with 

cattle: Journal of the American Veterinary Medicine 

Association, v. 215, no. 6, p. 792–794.

Schlottmann, J.L., Tanner, R., and Samadpour, M., 2000, 

Reconnaissance of hydrology, water quality, and 

sources of bacterial and nutrient contamination in the 

Ozark Plateaus aquifer system and Cave Springs 

Branch of Honey Creek, Delaware County, Oklahoma, 

March 1999–March 2000: U.S. Geological Survey 

Water-Resources Investigations Report 00–4210, 66 p.

Sullivan, A.E., Foster, D.I., Given, D., Einwalter, D.C., Graham, T., and Simpson, J., 1989, River use management 

plan—Ozark National Scenic Riverways: U.S. Department of the Interior, National Park Service, Denver Service Center, 46 p.

U.S. Environmental Protection Agency, 1985, Test methods 

for Escherichia coli and enterococci in water by the 

membrane filter procedure: U.S. Environmental Protection Agency, Cincinnati, Ohio, EPA 600/4-85-076, 

24 p.

——— 1986, Ambient water quality criteria for bacteria—1986: U.S. Environmental Protection Agency, 

Office of Water Regulations and Standards Division, 

Washington, D.C., EPA 440/5-84-002, 24 p.

University of Missouri, 2001, Population for Missouri counties, municipalities, and legislative districts 1990 and 

2000: Office of Social and Economic Data Analysis, 

Missouri State Census Data Center Program, 10 p., 

accessed May 2001, at URL http://mcdc.missouri.edu/census2000/pl94171.html.

Vandike, J.E., 1995, Surface water resources of Missouri—Missouri state water plan series, volume 1: 

Rolla, Missouri Department of Natural Resources, Geological Survey and Resource Assessment Division, 

Water Resources Report 45, 122 p.

Wilde, F.D., and Radtke, D.B., eds., 1998, National field 

manual for the collection of water-quality data—Field 

measurements: U.S. Geological Survey Techniques of 

Water-Resources Investigations, book 9, chap. A6, 

238 p.

Wilde, F.D., Radtke, D.B., Gibs, J., and Iwatsubo, R.T., eds., 

1999a, National field manual for the collection of 

water-quality data—Collection of water samples: U.S. 

Geological Survey Techniques of Water-Resources 

Investigations, book 9, chap. A4, 103 p.

——— 1999b, National field manual for the collection of 

water-quality data—Processing of water samples: U.S. 

Geological Survey Techniques of Water-Resources 

Investigations, book 9, chap. A5, 128 p.

Zaugg, S.D., Smith, S.G., Schroeder, M.P., Barber L.B., and 

Burkhardt, M.R., 2002, Methods of analysis by the U.S. 

Geological Survey National Water Quality Laboratory—Determination of wastewater compounds by 

polystyrene-divinylbenzene solid-phase extraction and 

capillary-column gas chromatography/mass spectrometry: U.S. Geological Survey Water-Resources Investigations Report 01–4186, 37 p.