TABLE OF CONTENTS

Chapter 1: Introduction....................................................................................... 1

1.1 Overview..................................................................................................... 1

1.2 Study Area.................................................................................................... 1

1.3 Sample Data and Collection Dates............................................................... 14

1.4 Baseflow Conditions................................................................................... 14

Chapter 2: Data Collection and Analysis............................................................ 25

2.1 Physical/Chemical Field Data....................................................................... 25

2.2 Herbicide Indicators.................................................................................... 36

2.3 Herbicide Samples....................................................................................... 37

2.4 Bacteriological Indicators............................................................................. 40

2.4.1 Total Coliforms................................................................................ 40

2.4.2 Fecal Coliforms............................................................................... 41

2.4.3 Escherichia coli................................................................................ 41

2.4.4 E. coli/Fecal Coliform Ratio............................................................. 42

2.4.5 Fecal streptococci........................................................................... 42

2.4.6 Enterococci..................................................................................... 42

2.5 Bacteriological Sampling.............................................................................. 42

2.5.1 Synoptic Fecal Coliform.................................................................. 43

2.5.2 Follow-Up Fecal Coliform............................................................... 43

2.6 Physical/Chemical Sampling......................................................................... 56

2.7 Nutrients..................................................................................................... 63

2.8 Nutrient Sampling........................................................................................ 65

2.9 Metals Data................................................................................................. 71

Chapter 3: Executive Summary......................................................................... 79

Chapter 4: Focused Sampling for Fecal Coliform............................................... 81

4.1 Clarks Run.................................................................................................. 81

4.2 Glenn’s Creek............................................................................................. 84

4.3 Herrington Lake.......................................................................................... 87

4.4 Hickman Creek and West Hickman Creek................................................... 90

4.5 McConnell Springs...................................................................................... 94

4.6 South Elkhorn Creek and Town Branch....................................................... 96

References............................................................................................................ 100

Appendix A: Quality Assurance and Quality Control

LIST OF TABLES

Table 1.1 2003 Kentucky River Watershed Watch Sampling Sites.................... 8

Table 1.2 Basinwide Sampling Data and Collection Dates............................... 14

Table 1.3 Types and Number of Samples at Sampling Sites............................ 15

Table 2.1 Physical/Chemical Field Data.......................................................... 26

Table 2.2 Herbicide Sampling Results............................................................. 40

Table 2.3 Synoptic Fecal Coliform Sampling Results....................................... 45

(by station identification number)

Table 2.4 Synoptic Fecal Coliform Sampling Results....................................... 49

(by concentration level)

Table 2.5 Follow-Up Fecal Coliform Sampling Results................................... 54

Table 2.6 Chemical Sampling Results.............................................................. 59

Table 2.7 Nutrient Sampling Results............................................................... 66

Table 2.8 Metals Sampling Results................................................................. 73

Table 2.9 Stations with Highest Metals Concentrations.................................... 78

LIST OF FIGURES

Figure 1.1 Kentucky River Basin........................................................................ 2

Figure 1.2 Kentucky River Basin 8-digit HUCs................................................... 3

Figure 1.3 Kentucky River Lower Basin............................................................. 4

Figure 1.4 Kentucky River Middle Basin............................................................ 5

Figure 1.5 Kentucky River Upper Basin............................................................. 6

Figure 1.6 Kentucky River Watershed Watch Monitoring Sites........................... 7

Figure 1.7 Kentucky River USGS Gaging Stations............................................ 21

Figure 1.8 North Fork Kentucky River............................................................. 22

Figure 1.9 Middle Fork Kentucky River........................................................... 22

Figure 1.10 South Fork Kentucky River............................................................. 23

Figure 1.11 Lock and Dam #10......................................................................... 23

Figure 1.12 Lock and Dam #2........................................................................... 24

Figure 2.1 Herbicide Sampling Locations.......................................................... 38

Figure 2.2 Kentucky River Basin Synoptic Fecal Coliform Counts..................... 44

Figure 2.3 Follow-Up Fecal Sampling Locations............................................... 53

Figure 2.4 Physical/Chemical Sampling Locations............................................. 58

Figure 2.5 Kentucky River Basin Phosphorus Sites > 1.0 mg/L......................... 69

Figure 2.6 Kentucky River Basin Nitrate Concentrations > 10 mg/L.................. 70

Figure 2.7 Kentucky River Basin Metal Sampling Locations.............................. 72

CHAPTER I: INTRODUCTION

1.1 Overview

This report documents the results of the 2003 Kentucky River Watershed Watch sampling effort, which was supported by grants from the Kentucky River Authority, Bluegrass PRIDE and Eastern Kentucky PRIDE. The Kentucky River Watershed Watch is a volunteer organization affiliated with the Kentucky Waterways Alliance with the following goals:

1. To provide current data on general water quality conditions to local stream based organizations working to protect their watershed.

2. To provide widespread screening for potential water quality problems to resource management agencies.

3. To provide auxiliary information to assist resource management agencies in meeting specific operational and management objectives.

4. To identify specific impacts to water quality through targeted observations and measurements.

The sampling effort was conducted so as to be consistent with the scientific study plan developed by the Kentucky River Watershed Watch scientific advisory board which describes the monitoring objectives, methods, parameters, quality assurance, and data management. A copy of the plan may be found at the project web site: http://water.nr.state.ky.us/watch/2000/plan_of_work.htm . Detailed sampling results for 2003 are posted on the project web site at http://nrepcapps.ky.gov/watch/management/ key.htm. All files associated with the Kentucky River basin begin with the letter “k.”

1.2 Study Area

The Kentucky River Watershed Watch sampling effort was conducted at 218 different sites across the Kentucky River Basin. The Kentucky River Basin extends over much of the central and eastern portions of the state and is home to approximately 710,000 Kentuckians. The watershed includes all or part of 42 counties and drains over 7,000 square miles with a tributary network of more than 15,000 miles. A map of the watershed with the associated counties is shown in Figure 1.1. For the purpose of watershed management, the River Basin has been subdivided into smaller sub-basins and watersheds using the USGS Hydrologic Unit Code (HUC) classification system. A map showing the 8-digit subbasins is shown in Figure 1.2. A more detailed description of the 11-digit HUC watersheds is provided in Figures 1.3-1.5. An index of the 218 sampling sites is provided in Figure 1.6 and Table 1.1.

1.3 Sample Data and Collection Dates

Water quality data were collected across the basin at four different times extending through the summer, and fall of 2003. A listing of the sample dates and types of data collected during each sample period is provided in Table 1.2. A summary of the types and number of samples collected at each data collection site is provided in Table 1.3.

Table 1.2 Basinwide Sample Data and Collection Dates

Type of Data Collected	Sample Dates	Sites	Samples
1. Herbicide	5/16/03-5/22/03	33	33
2. Synoptic Pathogens	7/11/03-7/14/03	151	151
3. Follow Up Pathogens	8/1/03 – 8/9/03	68	68
4a. Chemical/Nutrients	9/12/03-9/28/03	112	112
4b. Metals	9/13/03-9/21/03	39	39

1.4 Baseflow Conditions

In order to provide a basis for interpreting the sample results it is important to understand the associated stream conditions during the sampling effort. For example, data collected during low flow or dry conditions may be more indicative of the impact of points discharges while data collected following a storm may be more reflective of the impacts of non-point pollutant discharges. An indication of the stream conditions during the sampling period may be obtained by examination of USGS streamflow records. For the purposes of this study, five separate USGS gauging stations were selected for use in providing an indication of the streamflow conditions during the sampling period. The names, station numbers, and locations of each of these stations are shown in Figure 1.7. Streamflow plots for each station showing the times of the different sampling efforts are shown in Figures 1.8-1.12. (The streamflow values for these tables can be found on the USGS website at http://ky.water.usgs.gov.)

2.2 Herbicide Indicators

Two separate herbicides were used to evaluate the possibility of potential pollution from rural and/or urban land uses in the Kentucky River Basin. The herbicides included Metolachlor and Triazine.

Metolachlor is usually applied to crops before plants emerge from the soil, and is used to control certain broadleaf and annual grassy weeds in field corn, soybeans, peanuts, grain sorghum, potatoes, cotton, safflower, stone fruits, nut trees, highway right-of-ways and woody ornamentals. It inhibits protein synthesis; thus high protein crops (e.g. soy) can be adversely affected by excessive Metolachlor application. Additives may be included in product formulations to help protect sensitive crops (i.e. sorghum) from injury. Metolachlor is highly persistent in water over a wide range of acidity. At 20^◦ Celsius, its half-life is greater than 200 days in highly acidic water and is 97 days in highly basic water. Metolachlor is moderately persistent in the soil environment, with observed half-lives of 15 to 70 days. Breakdown rates are mainly dependent on microbial activity, and are therefore temperature-dependent. Metolachlor is currently unregulated by the U.S. Environmental Protection Agency, and therefore is not assigned a maximum contaminant level.

Triazine (or Atrazine) is a selective triazine herbicide used to control broadleaf and grassy weeds in corn and other crops, and in conifer reforestation plantings. It is also used as a nonselective herbicide on non-cropped industrial lands and on fallow lands. Over 64 million acres of cropland were treated with atrazine in the U.S. in 1990. Atrazine is moderately soluble in water. The main route of breakdown is chemical hydrolysis, followed by biodegradation. Atrazine is highly persistent in soil. Chemical hydrolysis followed by microbial breakdown accounts for most of its degradation in soil. Although hydrolysis is rapid in acidic or basic soil environments, it is slower at neutral pHs. The EPA’s drinking water standard maximum contaminant level for Atrazine is 0.003 mg/L (http://www.epa.gov/safewater/mcl.html). EPA's Office of Water has published a draft ambient water quality criteria document for atrazine containing acute and chronic criteria recommendations for the protection of aquatic life in both freshwater and saltwater. The procedures described in the "Guidelines for Deriving Numerical National Water Quality Criteria for the Protection of Aquatic Organisms and Their Uses" indicate that, except possibly where a locally important species is very sensitive, freshwater aquatic life and their uses should not be affected unacceptably if the one-hour average concentration does not exceed 350 ug/L more than once every three years on the average (acute criterion) and if the four-day average concentration of atrazine does not exceed 12 ug/L more than once every three years on the average (chronic criterion).

The basic manufacturer of both herbicides, Metolachlor and Atrazine, is Ciba-Geigy Agricultural Division. They can be contacted at the following address, phone number or fax number: Ciba-Geigy Agricultural Division; P.O. Box 18300; Greensboro, NC 27419-8300; Telephone: (919)632-6000; Fax: (919)299-8318.

2.3 Herbicide Samples

Herbicide data were collected at 33 sites during the period 5/16/03 – 5/22/03. Each sample was analyzed for both Metolachlor and Triazine. The location of each site is shown in Figure 2.1. A summary of the results for the herbicide data collection effort is provided in Table 2.2. Seventeen of the 33 sites had detectable levels of one or both of these herbicides, with Triazine registering more often. Site K263 (North Elkhorn Creek at Avon tributary) showed the highest value for Triazine, and site K260 (Dreaming Creek behind Madison Central High) showed the highest value for Metolachlor. None of the samples registering a detectable level of Atrazine exhibited a concentration greater than the EPA’s maximum contaminant level (MCL) of 0.003 mg/L or the EPA’s proposed criteria for aquatic life. Detectable levels for Atrazine ranged from a concentration of 0.08 ug/L to 1.49 ug/L.

2.4 Bacteriological Indicators

A number of pathogenic (disease causing) viruses, bacteria, and protozoans can enter a water body via fecal contamination. Human illness can result from drinking water or swimming in water that contains pathogens, or from eating shellfish harvested from such waters.

Unfortunately, direct testing for pathogens is impractical. Pathogens are rarely present in large numbers, and many are difficult to cultivate in the lab. Instead, microbiologists look for “inidator” species – so called because their presence indicates that fecal contamination may have occurred. The indicators most commonly used today include: total coliforms, fecal coliforms, Escherichia coli, fecal streptococci, and enterococci. Each of these bacteria are normally prevalent in the intestines and feces of warm-blooded animals, including humans. The indicator bacteria themselves are not usually pathogenic. All but E. coli are composed of a number of species of bacteria that share commons characteristics such as shape, habitat, or behavior; E. coli is a single species in the fecal coliform group.

There are basically two methods for analyzing water samples for bacteria:

The Membrane Filter Method involves filtering several different-sized portions of the sample using filters with a standard diameter and pore size, placing each filter on a selective nutrient medium in a Petri plate, incubating the plates at a specific temperature for a specified time period, and then counting the colonies that have grown on the filter. This method varies for different bacteria types (variations might include, for example, the nutrient medium type, the number and types of incubations, the method of incubations, etc.)

The Multipe-Tube Fermentation Method involves adding specified quantities of the sample to tubes containing a nutrient broth, incubating the tubes at a specified temperature for a specified time period, and then looking for the development of gas and/or turbidity that the bacteria produce. The presence or absence of gas in each tube is used to calculate an index known as the Most Probable Number (MPN).

2.4.1 Total Coliforms

Total coliforms are a group of bacteria that are widespread in nature. All members of the fecal coliform group can occur in human feces, but some can also be present in animal manure, soil, and submerged wood and in other places outside the human body. Thus, the usefulness of total coliforms as an indicator of fecal contamination depends on the extent to which the bacteria species found are fecal and human in origin. For recreational waters, total coliforms are no longer recommended as an indicator. For drinking water, total coliforms are still the standard test because their presence indicates contamination of a water supply by an outside source. Total coliforms are indicated in the lab by their ability to metabolize (ferment) the sugar lactose in an incubator at a temperature of 35C.

2.4.2. Fecal Coliforms

Fecal coliforms, a subset of total coliform bacteria, are more fecal specific in origin. However, even this group contains a genus, Lebsiella, with species that are not necessarily fecal in origin. Klebsiella are commonly associated with textile and pulp and paper mill wastes. Therefore, if these sources discharge to your stream, you might want to consider monitoring more fecal and human-specific bacteria. For recreational waters, this group was the primary bacteria indicator until relatively recently, when EPA began recommending E. coli and enterococci as better indicators of health risk from water contact. However, fecal coliforms are still being used in many states, including Kentucky, as the indicator bacteria. Similar to total coliforms, fecal coliforms are indicated in the lab by their ability to metabolize (ferment) the sugar lactose in an incubator at a temperature of 44.5 C. The state criteria for fecal coliform are based on the designated use of the particular stream and may be summarized as follows:

Primary Contact Recreation (swimming from May 1 thru Oct 31): fecal coliform shall not exceed 200 colonies per 100 ml as a monthly geometric mean based on not less than 5 samples per month; nor exceed 400 colonies per 100 ml in 20 percent or more of all samples taken during the month. [Note: As a result of the sampling frequency requirement with the first criteria, the state of Kentucky uses the 400 colonies per 100-ml criteria for classifying streams in the 305(b) report].

Secondary Contact Recreation (fishing and boating): fecal coliform content shall not exceed 1000 colonies per 100 ml as a monthly geometric mean based on not less than 5 samples per month; nor exceed 2000 colonies per 100 ml in 20 percent or more of all samples taken during the month.

Domestic Water Supply: fecal coliform content shall not exceed 2000 colonies per 100 ml as a monthly geometric mean based on not less than 5 samples per month.

2.4.3 Escherichia coli (E. coli)

The bacterium, E. coli, is commonly found in the intestines of healthy humans and animals and produces the K and B-complex vitamins that are then absorbed for nutritional benefit. The presence of E. coli in water indicates fecal contamination and the potential for waterborne disease. EPA recommends E. Coli as the best indicator of health risk from water contact in recreational waters. Kentucky is currently in the process of transitioning from a fecal coliform standard to an E. coli standard.

Although E. coli bacteria are not typically pathogenic, it has been found that E. coli concentrations are a predictor of swimming-associated gastrointestinal illness.* EPA bacterial water quality standards are thus based on a threshold concentration of E. coli in water above which the health risk from waterborne illness is unacceptably high. The EPA’s recommended water quality standard for recreational waters is based on 1) a geometric mean of 126 organisms/100 ml based on five samples collected during dry weather conditions or 2) 235 organisms/100 ml for any single water sample (EPA 1986).

* Dufour, Alfred P. 1984. Health effects criteria for fresh recreational waters. EPA-600/1-84-004. Office of Research and Development, USEPA, Washington, DC.

2.4.4 E. coli/Fecal Coliform Ratio

Because of the fact that E. Coli represent a sub-population of the fecal coliform bacteria, one should normally expect the ratio of the E. Coli to Fecal Coliform to be less than one. At a minimum, the E. coli test provides a second method for assessing the impairment of the water and the relative reliability of the fecal coliform count. However, depending on how the samples are analyzed it is possible to obtain ratios greater than one. In the current study, E. Coli were determined using a most probable number MPN technique while the fecal coliforms were determined using a membrane filtration method. In general, if the fecal coliforms are under biological stress, then is possible that some of them may actually die during the membrane filter test and not show up on the membrane filter. Because of the nature of the test, fewer organisms are susceptible to death using the MPN technique. As a consequence, monitoring sites which yield a EC/FC ratio greater than one, may be indicative of sites where the pathogens are under stress, such as a situation associated with some level of treatment (e.g. wastewater treatment plant, package plant, septic system).

2.4.5 Fecal Streptococci

Fecal streptococci generally occur in the digestive systems of humans and other warm-blooded animals. In the past, fecal streptococci were monitored together with fecal coliforms and a ratio of fecal coliforms to streptococci was calculated. This ratio was then used to determine whether the contamination was of human or nonhuman origin. However, because of the number of restrictions required for an accurate application of the ratio, this test is usually no longer recommended. Kentucky DOW currently does not employ fecal streptococci as a pathogen indicator for Kentucky watersways.

2.4.6 Enterococci

Enterococci are a subgroup within the fecal streptococcus group. Enterococci are distinguished by their ability to survive in salt water, and in this respect, they more closely mimic many pathogens than do the other indicators. Enterococci are typically more human-specific than the larger fecal streptococcus group. EPA currently recommends the use of Enterococci for testing marine recreational waters because of their superior correlation with swimming related illness. Kentucky DOW currently does not employ enterococci as a pathogen indicator for Kentucky waterways.

2.5 Bacteriological Sampling

Two different sets of fecal coliform sampling were conducted in the Kentucky River basin during the summer of 2003. These included synoptic sampling and follow-up sampling. The results of each sampling effort are discussed in following sections. During the first test all samples were analyzed for fecal coliforms using the membrane filter test. During the follow-up testing, all samples were analyzed again for fecal coliform using the membrane filter test as well as E. coli using an MPN method.

2.5.1 Synoptic Fecal Coliform Sampling

As in past years, a synoptic round of fecal coliform samples was collected at targeted sample locations during the month of July. The sample locations and associated results are shown in Figure 2.2. The individual results for each site are shown in Table 2.3. A ranking of the stations by the magnitude of the results is shown in Table 2.4

2.5.2 Follow-Up Fecal Coliform Sampling

Based on the observation of high readings at 103 of the synoptic sites (i.e., >400 CFU/100 ml), an additional round of fecal coliform sampling was conducted between 8/1/2003 and 8/9/2003. In addition to fecal coliform analyses, the samples were also evaluated for E. Coli. The sample locations and associated values are shown in Figure 2.3. The results of this sampling effort are provided in Table 2.5. A summary of the resulting ratios is provided in Table 2.6. Results indicated continuing fecal-related problems at 48 of the 61 re-sampled sites (concentrations greater than 400 fecal colonies/100 ml).

2.6 Physical/Chemical Sampling

General chemical data (alkalinity, chlorides, conductivity, total suspended solids, and total hardness) were collected at all sample locations during the month of September. The locations of the sampling sites are shown in Figure 2.4. The individual results for each sample are shown in Table 2.7.

Estradiol: This steroidal estrogen hormone is used in women’s hormone replacement drugs, as well as drugs given to livestock. Main sources of high concentrations of estradiol in water are sewage treatment wastewater (including septic systems) or livestock waste. As a result, the presence of estradiol would indicate the presence of human sewage or runoff of livestock waste. Estradiol can lead to reproductive impairment, endocrine disruption, and death in fish populations. Its effects on human populations are still being studied.

Alkalinity: Alkalinity refers to the degree to which the water sample is basic, or has a pH greater than 7, and affects the capability of water to neutralize acid. In most natural water bodies in Kentucky the buffering system is carbonate-bicarbonate. Alkalinity is important for fish and aquatic life because it protects or buffers against rapid pH changes. Higher alkalinity levels in surface waters will buffer acid rain and other acid wastes and prevent pH changes that are harmful to aquatic life. Kentucky’s water quality criteria state that for protection of aquatic life, the buffering capacity should be at least 20 mg/L. If alkalinity is naturally low, (less than 20 mg/L) there can be no greater than a 25% reduction in alkalinity.

Chlorides: Chlorides are salts resulting from the combination of the gas chlorine with a metal. Fish and aquatic communities cannot survive in waters with high levels of chlorides. Public Drinking Water Standards require chloride levels not to exceed 250 mg/L. Criteria for protection of aquatic life require levels of less than 600 mg/L for chronic (long-term) exposure and 1200 mg/L for short-term exposure.

Conductivity: Conductivity is a measurement of the ability of an aqueous solution to carry an electrical current. Conductivity measurements are used to determine mineralization, or total dissolved solids. Indirect effects of excess dissolved solids are primarily the elimination of desirable food plants and habitat-forming plant species. For Kentucky, water quality criteria have been established only for the mainstem of the Ohio River. The limit is 800 micromhos/cm or 500 mg/L total dissolved solids.

Total Suspended Solids: One of the biggest sources of water pollution in Kentucky is suspended solids. Suspended solids include inorganic particles (silts, clays, etc.) and organic particles (algae, zooplankton, bacteria, and detritus) that are carried along by water as it runs off the land. The inorganic portion is usually considerably higher than the organic. Both contribute to turbidity, or cloudiness of the water. High values of TSS cause multiple environmental impacts, including clogging fish gills, reducing light penetration, and siltation of stream bottoms and associated habitats. Indirectly, the suspended solids affect other parameters such as temperature and dissolved oxygen. Suspended solids also interfere with effective drinking water treatment. High sediment loads interfere with coagulation, filtration, and disinfection, and more chlorine is required to effectively disinfect turbid water.

There are no quantitative criteria for TSS; however, Kentucky Water Quality Standards for aquatic life state that suspended solids "shall not be changed to the extent that the indigenous aquatic community is adversely affected" and "the addition of settleable solids that may adversely alter the stream bottom is prohibited."

Total Hardness: Hardness is due to the presence of multivalent metal ions which come from minerals dissolved in the water. Generally, harder water results in a lower toxicity of other metals to aquatic life. In fresh water the primary ions are calcium and magnesium; however iron and manganese may also contribute. There are no Kentucky water quality criteria for hardness.