In cooperation with the West Virginia Department of Environmental Protection, 

Division of Water and Waste Management and the West Virginia Division of Natural 

Resources 

A Reconnaissance for Emerging Contaminants in the 

South Branch Potomac River, Cacapon River, and 

Williams River Basins, West Virginia, April-October 

2004 

By Douglas B. Chambers and Thomas J. Leiker 

Open-File Report 2006-1393 

U.S. Department of the Interior 

U.S. Geological Survey 

i 
U.S. Department of the Interior 

Dirk Kempthorne, Secretary 

U.S. Geological Survey 

Mark D. Myers, Director 

U.S. Geological Survey, Reston, Virginia 2006 

For product and ordering information: 

World Wide Web: http://www.usgs.gov/pubprod

Telephone: 1-888-ASK-USGS 

For more information on the USGS—the Federal source for science about the Earth, 

its natural and living resources, natural hazards, and the environment: 

World Wide Web: http://www.usgs.gov 

Telephone: 1-888-ASK-USGS 

Suggested citation: 

Chambers, D.B., and Leiker, T. J., 2006, A Reconnaissance for Emerging Contaminants in the South 

Branch Potomac River, Cacapon River, and Williams River Basins, West Virginia, April-October 

2004: U.S. Geological Survey Open-File Report 2006-1393, 23 p. http://pubs.usgs.gov/of/2006/1393 

Any use of trade, product, or firm names is for descriptive purposes only and does not imply 

endorsement by the U.S. Government. 

Although this report is in the public domain, permission must be secured from the individual 

copyright owners to reproduce any copyrighted material contained within this report. 

ii
Contents 

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

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

Purpose and Scope ........................................................................................................................................... 2 

Description of Study Area.................................................................................................................................... 3 

Potential Sources of Contaminants................................................................................................................ 4 

Agricultural Runoff......................................................................................................................................... 4 

Point-Source Pollution.................................................................................................................................. 5 

Methods .................................................................................................................................................................. 6 

Passive Samplers .............................................................................................................................................. 6 

SPMD ............................................................................................................................................................... 6 

POCIS ............................................................................................................................................................... 8 

Passive-Sampler Deployment ..................................................................................................................... 8 

Stream-Water and Effluent Sampling............................................................................................................ 9 

Blood-Plasma Collection.................................................................................................................................. 9 

Analytical Methods ............................................................................................................................................. 10 

Passive-Sampler Extracts.............................................................................................................................. 10 

Blood Plasma.................................................................................................................................................... 12 

Antibiotics ......................................................................................................................................................... 12 

Roxarsone and Other Arsenic Species........................................................................................................ 12 

Results ................................................................................................................................................................... 12 

Passive-Sampler Results................................................................................................................................ 12 

Blood-Plasma Results..................................................................................................................................... 15 

Water-Sample Results.................................................................................................................................... 15 

Summary ............................................................................................................................................................... 18 

References Cited ................................................................................................................................................. 20

Figures 

1. Map showing emerging contaminant reconnaissance study areas in the South Branch Potomac 

River, Cacapon River, and Williams River Basins...........................................................................................3 

2. Map showing locations of sites in the South Branch Potomac River, Cacapon River, and Williams 

River Basins where stream water, wastewater effluent, or smallmouth bass blood plasma 

were collected or passive samplers deployed, April-October 2004............................................................8 

iii
Tables

1. Media sampled at emerging-contaminant reconnaissance sites in the South Branch Potomac 

River, Cacapon River, and Williams River Basins, April-October 2004.......................................................7 

2. Chemical compounds analyzed for in passive-sampler (SPMD and POCIS) extracts and 

associated estimated detection levels ...........................................................................................................11 

3. Chemical compounds analyzed for in smallmouth bass blood-plasma samples and associated 

detection levels ...................................................................................................................................................13 

4. Antibiotics analyzed for in passive sampler extracts and in wastewater effluent in the South 

Branch Potomac River, Cacapon River, and Williams River Basins .........................................................14 

5. Chemical compounds in passive samplers (SPMD and POCIS) deployed at sites in the South 

Branch Potomac River, Cacapon River, and Williams River Basins, April to June 2004.......................16 

6. Chemical compounds in smallmouth bass blood-plasma samples from the South Branch Potomac 

River and the Williams River Basins, May to October 2004 ........................................................................17 

7. Inorganic arsenic and organoarsenic compounds in stream-water and effluent samples collected 

at sites in the South Branch Potomac River and Cacapon River Basins, September 2004 ..................19 

8. Antibiotics and antibiotic degradation products in composite samples collected from wastewater 

effluent in the South Branch Potomac River Basin, June 7-8, 2004 ..........................................................19 

iv
Conversion Factors 

Inch/Pound to SI 

Multiply By To obtain

Length 

foot (ft) 0.3048 meter (m) 

mile (mi) 1.609 kilometer (km)

Area 

acre 0.4047 hectare (ha)

square mile (mi

2

) 259.0 hectare (ha)

square mile (mi

2

) 2.590 square kilometer (km2) 

Volume 

gallon (gal) 3.785 liter (L) 

cubic foot (ft3

) 0.02832 cubic meter (m3) 

cubic yard (yd

3

) 0.7646 cubic meter (m3) 

Mass 

ounce, avoirdupois (oz) 28.35 gram (g) 

pound, avoirdupois (lb) 0.4536 kilogram (kg)

Pressure 

inch mercury (in Hg) 25.4 Torr

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

°F=(1.8×°C)+32

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

°C=(°F-32)/1.8 

Horizontal coordinate information is referenced to the North American Datum of 1983 (NAD 83). 

v
A Reconnaissance for Emerging Contaminants in the 

South Branch Potomac River, Cacapon River, and 

Williams River Basins, West Virginia, April-October 

2004 

By Douglas B. Chambers, and Thomas J. Leiker

Abstract 

In 2003 a team of scientists from West 

Virginia Division of Natural Resources and 

the U. S. Geological Survey found a high 

incidence of an intersex condition, oocytes in 

the testes, among smallmouth bass 

(Micropterus dolomieu) in the South Branch 

Potomac River and the Cacapon River of 

West Virginia, indicating the possible 

presence of endocrine-disrupting compounds 

(EDCs). Possible sources of EDCs include 

municipal and domestic wastewater, and 

agricultural and industrial activities. Several 

sampling strategies were used to identify 

emerging contaminants, including potential 

EDCs, and their possible sources in these 

river basins and at an out-of-basin reference 

site. Passive water-sampling devices, which 

accumulate in-stream organic chemical 

compounds, were deployed for 40-41 days at 

8 sampling sites. Sampler extracts were 

analyzed for a broad range of polar and nonpolar organic compounds including 

pesticides, flame retardants, pharmaceuticals, 

and personal-care products. Analysis of 

passive-sampler extracts found 4 compounds; 

hexachloro-benzene; pentachloroanisole; 

2,2',4,4',5-penta-bromo-diphenyl ether (BDE 

47); and 2,2',4,4',6-penta-bromo-diphenyl 

ether (BDE 99) to be present at every 

sampled site, including the reference site, and 

several sites had detectable quantities of other 

compounds. No detectable quantity of any 

antibiotics was found in any passive-sampler 

extract. Effluent samples were analyzed for 

39 antibiotics as tracers of human and 

agricultural waste. Additionally, poultryprocessing plant effluent was sampled for 

roxarsone, an organoarsenic compound used 

as a poultry-feed additive, and other arsenic 

species as tracers of poultry waste. 

Antibiotics were detected in municipal 

wastewater, aquaculture, and poultryprocessing effluent, with the highest number 

of antibiotics and the greatest concentrations 

found in municipal effluent. Arsenate was 

the only arsenic species detected in the 

poultry-processing plant effluent, at a

concentration of 1.0 μg/L. Water samples 

were collected from 7 stream sites and 

analyzed for arsenic species, including

roxarsone. Arsenate was detected in samples 

from 6 of the 7 stream samples, in 

concentrations ranging from 0.3 to 0.5 μg/L. 

Additionally, the analysis of smallmouth bass 

blood plasma for potential EDCs indicated 

the presence of several compounds including 

some found in the passive sampler extracts, 

specifically BDE 47 and BDE 99. Data from 

this reconnaissance will help to focus efforts 

for further studies of the occurrence of 

emerging contaminants, EDCs, and intersex 

in smallmouth bass in these Potomac River 

tributaries. 

1
Introduction 

In 2003 West Virginia Division of 

Natural Resources (WVDNR) officials and 

United States Geological Survey (USGS) 

Leetown Science Center scientists examining 

fish kills and widespread incidences of fish 

lesions in the South Branch Potomac River 

(fig. 1), discovered many reproductive 

anomalies among smallmouth bass 

(Micropterus dolomieu) (Vicki Blazer, 

USGS, written communication, 2004). These 

anomalies were both widespread and 

pervasive within communities and included 

oocytes, cells that are precursors to eggs, in 

the testes of male fish. Abnormal 

development of reproductive tissues may 

result from disruptions of the endocrine 

system brought about by exposure to 

anthropogenic sources of hormones and 

synthetic compounds that mimic hormones 

including ethynyl estradiol, nonyl phenol, 

polybrominated diphenyl ethers. Earlier 

studies by USGS researchers had found 

feminization of male common carp (Cyprinus 

carpio) in the Shenandoah River near 

Millville, WV, another tributary of the 

Potomac River (Goodbred and others, 1997). 

Several classes of synthetic organic 

compounds to abnormal development or 

disturbances of the endocrine system in fish 

and other aquatic vertebrates (Jobling and 

others, 2002; Sheahan and others, 2002; 

Orlando and others, 2004; Nash and others, 

2004; Scheurs and others, 2004; Melo and 

Ransdell, 2001; Kavanagh and others, 2004; 

Toft and others, 2003; Bevan and others, 

2003). These compounds, known as 

endocrine-disrupting compounds (EDCs), 

mimic natural hormones and interfere with an 

organism’s internal signaling and regulatory 

systems. Excess quantities of natural 

hormones, derived from people or livestock, 

can also enter the environment and potentially 

effect aquatic organisms (Nichols and others, 

1997; Dyer and others, 2001; Carballa and 

others, 2004; Hutchison and others, 2005; 

Raman and others, 2004; Hanselman and 

others, 2003). EDCs and natural hormones 

can enter the environment through 

agricultural, industrial, municipal, and 

domestic wastes (Kolpin and others, 2002, 

Kolodziej and others, 2004). 

Histopathologic examinations of 

smallmouth bass testes collected from the 

South Branch Potomac, Potomac, and 

Cacapon Rivers in 2004 (fig. 1) found an 

intersex condition, oocytes in the testes, 

affecting about 80% of the male fish sampled 

(Vicki Blazer, USGS, written 

communication, 2004). While there is no 

widely accepted normal rate of intersex 

occurrence, it is clear that observed rates 

exceed expected values and may be due to the 

presence of hormones or other EDCs in the 

environment. In 2004 the USGS, in 

cooperation with the West Virginia 

Department of Environmental Protection, 

Division of Water and Waster Management 

and the West Virginia Division of Natural 

Resources, initiated a study to characterize 

the chemical environment of the South 

Branch Potomac River and the Cacapon River 

Basins, with a reference site in the Williams 

River Basin, all in West Virginia (fig. 1). 

This study collected and analyzed water and 

smallmouth bass blood-plasma samples, and 

used innovative passive sampling devices 

(Alvarez and others, 2004) to identify 

emerging contaminants, including EDCs, 

which may be present in these watersheds. 

Purpose and Scope 

The purpose of this report is to present 

results of the analyses of passive-sampler 

extracts, stream-water samples, wastewater 

effluent samples, and smallmouth bass bloodplasma samples from sites in the South 

Branch Potomac River, the Cacapon River, 

and Williams River Basins of West Virginia 

(fig. 1) in 2004. These samples were 

2 
Figure 1. Emerging contaminant reconnaissance study areas in the South Branch Potomac River, 

Cacapon River, and Williams River Basins. 

collected to detect emerging contaminants, 

including possible EDCs that may result in 

increased intersex in native male smallmouth 

bass. 

Passive samplers, both semipermeable 

membrane devices (SPMDs) and polar 

organic compound integrative samplers 

(POCIS) (Alvarez and others, 2004; Petty and 

others, 2004), were deployed at six sites in 

the Potomac River Basin, one site in the 

Cacapon River Basin, and one reference site 

in the Williams River Basin. Extracts derived 

from the passive samplers were analyzed for 

several classes of emerging contaminants 

including antibiotics and personal-care 

products. Stream-water samples were 

analyzed for both organic and inorganic 

species of arsenic. All samples collected 

from wastewater effluents were analyzed for 

antibiotics, and one also was analyzed for 

arsenic species. Blood-plasma samples 

collected from smallmouth bass were

analyzed for several classes of emerging 

contaminants including personal-care 

products. 

Description of Study Area 

The study area includes the South 

Branch Potomac River and Cacapon River 

Basins located in the Eastern Panhandle of 

West Virginia, and the Williams River Basin 

in central West Virginia (fig. 1). The initial 

discovery of intersex condition in smallmouth 

bass in the Potomac River Basin was in the 

3 
South Branch Potomac and the Cacapon 

River Basins (Vicki Blazer, USGS, written 

communication, 2004). The Williams River 

site was selected as an out-of-basin reference 

condition. 

The South Branch Potomac River 

drains 1,506 mi2

 of West Virginia and 

Virginia. Population density is low, with an 

estimated mean population density of 20 

people per mi2

, and an estimated total 

population in 2000 of approximately 30,100 

(U.S. Census Bureau, 2002). The largest 

communities in the basin are Moorefield, 

population 2,375; Petersburg, population 

2,423; Romney, population 1,940; and 

Franklin, population 797, all in West Virginia 

(U.S. Census Bureau, 2002). The area is 

rural and agricultural activities, primarily 

poultry, cattle and sheep rearing, are 

significant sources of income. Poultryrearing and poultry-processing operations are 

major employers. Other important 

agricultural activities include low-density 

cattle and sheep rearing. Tourism also plays 

a significant role in the economy. Many 

people travel to the area for outdoor 

recreation activities such as backpacking, 

rock climbing, and trout and smallmouth bass 

angling. 

The Cacapon River Basin (fig 1.) is 

adjacent to the South Branch Potomac River 

and has similar land uses and population 

densities; it drains 681 mi2

 of West Virginia’s 

Eastern Panhandle. Population density is 

low, with an estimated mean population 

density of 23 people per mi2

, and an estimated 

total population in 2000 of approximately 

15,500. The largest community in this basin 

is Great Cacapon, population 1,379 (U.S. 

Census Bureau, 2002). Poultry rearing is an 

important source of income. The highest 

density of poultry rearing houses in the study 

area is in the Lost River basin, an upper 

Cacapon River tributary (fig. 1). 

Samples were also collected from the 

Williams River at Dyer, WV. The Williams 

River is in the Kanawha River Basin of the 

Ohio River Basin, and drains 129 mi2

 of a 

remote portion of West Virginia. This site 

served as a comparative reference site for the 

Kanawha-New River NAWQA Study 

(Paybins and others, 2000) and was selected 

as an out-of-basin reference site for this 

study. There are no incorporated 

communities located in the Williams River 

drainage. The mean population density is less 

than 8 people per mi2

 and the estimated 

population is less than 1,000 (U.S. Census 

Bureau, 2002). The Williams River site was 

selected as a reference due to its size, the 

presence of native smallmouth bass, and lack 

of potential sources of contaminants. No site 

in the South Branch Potomac River Basin 

offered this combination of characteristics. 

Potential Sources of Contaminants

Many potential sources of

contaminants discharge to the South Branch 

of the Potomac and Cacapon Rivers. Chief 

among these are runoff from agricultural 

activities, municipal and domestic wastewater 

effluent (both treated and untreated),

industrial wastewater, and gypsy moth control 

programs using dimilin (diflubenzuron). 

Agricultural Runoff 

Livestock rearing is the principal 

agricultural activity in the South Branch 

Potomac and Cacapon River Basins. Chicken 

and turkey rearing are the principal livestock 

activities followed by low-intensity beef 

cattle rearing, with approximately 82,000 

cattle raised in the area, and a lesser amount 

of sheep rearing, with approximately 11,000 

sheep in the area. Crop production is limited; 

corn is the primary grain, with some wheat 

production; there are approximately 57,000 

acres and 3,700 acres of land in row crop 

production, respectively, in the South Branch 

Potomac and Cacapon River Basins. 

The Eastern Panhandle of West 

Virginia is a center of poultry production. 

According to the National Agricultural 

Statistics Service’s 2002 West Virginia 

Census of Agriculture (2004), more than 

90,000,000 birds were sold from the region in 

4 
2002 and approximately 450 chicken and 

turkey rearing facilities were found 

throughout the South Branch Potomac and 

Cacapon River Basins in 2002. When the 

birds reach market size they are transported to 

a processing plant for slaughter and 

packaging. Currently a poultry-processing 

plant operates in Moorefield, WV, in the 

South Branch Potomac River Basin that 

slaughters and processes most of the chickens 

raised in the area. Most of the turkeys reared 

in the area are slaughtered and processed at a 

facility in Harrisonburg, VA. After the birds 

have left the rearing house, the house litter (a 

mixture of pine shavings, excrement, and 

other residue) is removed from the house and 

composted. The composted litter is later 

spread on pastures and crop fields as both a 

fertilizer and method of waste disposal, with 

some composted litter trucked out of basin for 

land application elsewhere. 

Two primary concerns associated with 

runoff from poultry-litter applications are 

17β-estradiol and arsenic. The estradiol is a 

naturally occurring reproductive hormone that 

is shed in poultry feces and persists through 

composting (Nichols and others, 1997; 

Hanselman and others, 2003). Arsenic is 

introduced to the poultry as roxarsone, an 

organoarsenic compound originally used as 

anti-parasitic agent and now used to improve 

the color and texture of chicken and turkey 

meat (U. S. Food and Drug Administration, 

2000). 

Roxarsone does not accumulate in the 

poultry meat but is shed in the feces, the birds 

are taken off of roxarsone-amended feed five 

days before slaughter to reduce body burdens. 

Roxarsone and other arsenic species resulting 

from biotransformation of roxarsone are 

present in the composted poultry-house litter 

that is subsequently applied to pastures and 

crop fields. Studies of arsenic in a small 

agricultural basin indicate that much of this 

arsenic strongly sorbs to soils, but a fraction 

of the arsenic is washed into streams by storm 

events following the springtime application of 

poultry-house litter (Brown and others, 2005; 

Schreiber and others 2004). 

Antibiotics are another suite of 

compounds associated with poultry 

production, used for both disease treatment

and growth promotion. Macrolides 

(virginiamycin, lincomycin) and tetracyclines 

are the most extensively used antibiotic 

classes in poultry rearing. The primary 

release of poultry-rearing antibiotics to the 

environment is through the spreading of 

poultry-house litter on croplands and pastures 

(Chapman and Johnson, 2002; Campagnolo 

and others, 2002). 

Point-Source Pollution 

Wastewater effluent and other 

environmental releases from municipal, 

industrial and residential sources represent 

potentially significant inputs of contaminants 

in the South Branch Potomac and Cacapon 

River Basins. There are no point discharges 

permitted under the National Pollutant 

Discharge Elimination System (NPDES) or 

other regulated waste releases in the Williams 

River Basin (WVDEP, 2001, GIS coverage). 

In the South Branch Basin there are 

73 NPDES discharges permitted by West 

Virginia Department of Environmental 

Protection (WVDEP, 2001, GIS coverage). 

These permits are for industrial storm-water 

runoff, general sewage, aquaculture facilities, 

landfill leachate, and various other effluent 

types. The largest facilities are the 

wastewater-treatment plants for the towns of 

Moorefield, Romney, Franklin, and 

Petersburg, and the Cherry Grove U.S. Naval 

Station. 

The West Virginia Department of

Environmental Protection has granted 

NPDES permits to 24 discharges in the 

Cacapon River drainage for general sewage, 

industrial storm water runoff, sanitary landfill 

leachate, and various other discharges. The 

largest permitted discharges are wastewatertreatment plants for the towns of 

Wardensville and Capon Bridge, and East 

5 
6 

 

Hardy High School (WVDEP, 2001, GIS 

coverage). 

In the South Branch Potomac, 

Cacapon, and Williams River Basins, a large 

but unquantified portion of residential sewage 

is either treated by septic tank/leach field 

systems or discharged directly to surface 

waters without treatment. The “straight-pipe” 

untreated discharges are a problem 

throughout West Virginia (WVDEP, 2006). 

The South Branch Potomac River 

Basin is the only study basin with a 

significant amount of industry. This includes 

a polymer-compounding plant in Petersburg 

and poultry-feed mill, poultry-processing 

plant and cabinetry factory in Moorefield. 

Compounds emitted by these facilities, 

according to the 2002 USEPA toxic release 

inventory, include organic compounds such 

as xylene, toluene, methanol, and 

ethylbenzene, and inorganic compounds such 

as nitrates, lead, copper, and manganese 

compounds (U.S. Environmental Protection 

Agency Toxic Release Inventory, 2004). 

Methods 

A broad program of sampling, 

designed to account for several potential 

sources of emerging contaminants, including 

endocrine disrupting compounds, was 

initiated in late April 2004. Water quality 

was assessed through the use of passive 

samplers, the collection of water samples 

from streams and waste-water discharges, and 

the collection of smallmouth bass blood 

plasma (table 1). 

Passive Samplers 

Passive-sampling technology typically 

uses an encased media that can accumulate 

and sequester compounds of interest over a 

period of time (Alvarez and others, 2004). 

Passive-sampling approaches offer several 

advantages; chief among these are the ability 

to integrate exposure over time and a range of 

hydrologic conditions, and the ability to 

accumulate a detectable mass of a compound 

that may be present in a water sample at 

concentrations below the method detection 

level (Alvarez and others, 2005). Several 

passive-sampling methods have been tested 

and used in aquatic environments; HPLCgrade hexane contained in dialysis tubing for 

accumulating hydrophobic organic 

compounds, lipids encased in plastic tubing 

for hydrophobic organic compounds, and 

waxes that exude from tubing to sorb metals 

and other inorganic constituents. This study 

used semipermeable membrane devices 

(SPMDs) to accumulate hydrophobic organic 

compounds (Huckins and others, 2002) and 

polar organic compound integrative samplers 

(POCIS) (Alvarez and others, 2004) to 

accumulate hydrophilic organic compounds. 

SPMD 

SPMDs consist of a small volume of neutral 

synthetic lipid encased in polyethylene tubing 

(Huckins and others, 2002). The SPMDs are 

designed to maximize the surface area to 

volume ratio, thereby maximizing the uptake 

efficiency of the device. The SPMDs used in 

this study were made by Environmental 

Sampling Technologies, Inc. of St. Joseph, 

MO1

 and were 91.4 cm by 2.5 cm lay-flat 

polyethylene tubing made without additives 

and filled with ultra-high purity synthetic 

triolein, a high molecular weight neutral lipid. 

Dissolved phase hydrophobic organic 

compounds sorb to the surface of the SPMD. 

A fraction of the sorbed material will move 

through channels in the polyethylene 

membrane to the internal triolein phase, the 

remainder of the sorbed material will remain 

sorbed to the exterior of the membrane or be 

held in the transient channels through the 

membrane. 

1

 The use of tradenames is only for identification 

purposes and does not imply endorsement by the U. S. 

Geological Survey. 
7 

Table 1. Media sampled at emerging-contaminant reconnaissance sites in the South Branch Potomac River, Cacapon River, and Williams River Basins, AprilOctober 2004. [x, media sampled at each site; SPMD, semipermeable membrane device; POCIS, polar organic compound integrative sampler] 

Sample media collected and analyzed 

Water 

Site name Map 

index 

number 

(fig. 1) 

USGS Station 

identification 

number 

Passive 

Samplers 

(SPMD 

and 

POCIS) 

Antibiotics Arsenic species 

Smallmouth 

bass blood 

plasma 

South Branch Potomac River near Upper Tract, WV 1 384829079163401 x x x North Fork South Branch Potomac River at Cabins, WV 2 01606000 x x x South Branch Potomac River at Petersburg, WV 3 01606500 x Spring Run Hatchery effluent near Masonville, WV 4 385509079050601 x Mill Creek near Petersburg, WV 5 01607200 x Lunice Creek at Petersburg, WV 6 385938079064301 x Petersburg wastewater treatment plant discharge at Petersburg, WV 7 385930079064501 x South Branch Potomac River at Petersburg Gap, WV 8 390011079044501 x x South Fork South Branch Potomac River near Moorefield, WV 9 01608000 x Moorefield wastewater treatment plant discharge at Moorefield, WV 10 390424078582301 x SH discharge at Moorefield, WV 11 390329078583001 x x South Branch Potomac River near Moorefield, WV 12 01608070 x x South Branch Potomac River at railroad bridge at Sycamore, WV 13 390909078545201 x x South Branch Potomac River near Springfield, WV 14 01608500 x Lost River at McCauley near Baker, WV 15 01610200 x x Williams River at Dyer, WV 16 03186500 x x 
Figure 2. Locations of sites in the South Branch Potomac River, Cacapon River, and Williams River 

Basins where stream water, wastewater effluent, or smallmouth bass blood plasma were collected 

or passive samplers deployed, April-October 2004. Williams River Basin is shown in inset.

POCIS 

POCIS sequester polar organic 

compounds in a solid phase media encased in 

a porous membrane (Alvarez and others, 

2004). The POCIS consist of a mixed solid 

sequestration phase sealed in a hydrophilic 

microporous membrane. Water penetrates the 

encasing membrane and infiltrates the 

sequestration polymers, which sorb and 

immobilize water-soluble compounds 

throughout the exposure time. The POCIS 

devices used in this study were also 

manufactured by Environmental Sampling 

Technologies, Inc. 

Passive-Sampler Deployment 

Passive samplers were deployed at a 

total of eight sites, six stream sites in the 

South Branch Potomac River Basin, one site 

in the Cacapon River Basin, and one site in 

the Williams River Basin (table 1 and fig. 2). 

At each site three SPMDs and three POCIS 

were deployed in perforated stainless steel 

protective canisters. Care was taken to place 

the canisters where they would likely remain 

submerged for the duration of the deployment 

and in flowing water, avoiding backwater 

areas that may not be representative of main 

channel flow and water quality. Samplers 

were deployed at the South Branch Potomac 

8 
River near Springfield, near Moorefield, at 

Petersburg Gap, below Upper Tract, and in 

the North Fork South Branch Potomac River 

near Cabins during April 29-30, 2004 and 

retrieved June 7-8, 2004. Streamflow during 

deployment was typical for the season, 

moderately high flows punctuated with higher 

storm flows. Samplers were deployed at the 

Williams River at Dyer reference site on May 

20, 2004 and retrieved July 1, 2004, a period 

when springtime flows were receding to 

typical summertime flows. The South Branch 

Potomac River at Sycamore and Lost River at

Baker passive samplers were deployed on 

May 24-25, 2004 and retrieved on July 7-8, 

2004, again during a period when springtime 

flows are receding to summertime lows. At 

the end of the 39-44 days of in-stream 

deployment, the passive samplers were 

removed from the deployment canisters, 

rinsed with native water, placed in sealed 

containers, chilled, and transported to the 

USGS West Virginia Water Science Center 

laboratory. 

Because SPMDs can efficiently 

accumulate airborne contaminants, a tripblank SPMD was used at each site. An 

individually packed trip-blank SPMD was 

removed from its sealed container and 

exposed to ambient air for the time interval

during initial deployment when the sample 

SPMDs were exposed to the air, from 

removal from their sealed container to 

immersion in stream water. The trip-blank 

SPMDs were returned their sealed containers 

and kept until retrieval of the passive 

samplers, at which time the trip-blank

SPMDs were again exposed to ambient air, 

from the time the sample SPMDs were 

removed from the water until they were 

returned to their sealed containers. Two 

polybrominated diphenyl ethers, 2,2',4,4'-

tetra-bromo-diphenyl ether (BDE 47) and 

2,2',4,4',5-penta-bromo-diphenyl ether (BDE 

99), were found in trip blanks SPMD from 

three sites; South Branch Potomac River near 

Springfield (3.9 ng/g SPMD BDE 47 and 

4.87 ng/g SPMD BDE 99), at Sycamore (6.24 

ng/g SPMD BDE 47 and 6.26 ng/g SPMD 

BDE 99), and near Upper Tract (4.9 ng/g 

SPMD BDE 47 and 5.1 ng/g SPMD BDE 99). 

Stream-Water and Effluent Sampling 

Stream-water and effluent samples 

were collected at selected sites in the South 

Branch of the Potomac River and Cacapon 

River Basins; samples from four sites were 

analyzed for antibiotics and eight sites were 

sampled for arsenic species (table 1 and fig. 

2). Stream-water samples were collected by 

West Virginia Department of Environmental 

Protection (WVDEP) personnel from five 

equidistant points across a stream using a 

weighted-bottle sampler. WVDEP personnel 

received effluent discharge as composited 

samples from the plant operator. Samples 

collected for analysis of roxarsone and other 

arsenic species were filtered at 0.45 μm, and 

100 μL of EDTA solution was added to 

preserve ambient valence states of all arsenic 

species (Garbarino and others, 2000). 

Unfiltered samples were collected in 1-L 

amber glass bottles, chilled to 4° C, and 

shipped to the USGS Organic Geochemistry 

Research Laboratory in Lawrence, Kansas for 

analysis of antibiotics. 

Blood-Plasma Collection 

Smallmouth bass were collected by 

electrofishing using USGS Biomonitoring of 

Environmental Status and Trends (BEST) 

protocols (Schmitt and others, 1999) at 5 sites 

in the South Branch Potomac and one site at 

the Williams River at Dyer (table 1 and fig. 

2). Captured smallmouth bass were weighed, 

measured, and sacrificed. Blood was 

collected from the caudal artery using a 5-mL 

syringe. Withdrawn blood was transferred to 

5-mL heparinized vacutainers and centrifuged 

for 10 min. at 1,000 x g (where the constant g 

is the acceleration due to gravity). 

Supernatant plasma was transferred to 

microcentrifuge tubes and stored at -70°C 

until analysis. A subset of samples, blood 

plasma from 30 individuals from six sites, 

9 
was selected for analysis based on fish gender 

and degree of intersex condition. 

Analytical Methods 

Passive-Sampler Extracts 

Extracts for SPMD and POCIS 

devices are prepared using different 

techniques. SPMD were extracted through 

dialysis of the entire SPMD into pesticidegrade hexane. Hydrophobic compounds 

preferentially partition into the dialysis 

solvent, the dialysis solvent now containing 

the hydrophobic compounds is the SPMD 

extract. POCIS extracts were prepared by 

eluting hydrophilic compounds from the 

POCIS sequestration medium with HPLCgrade methanol. Following dialysis or 

elution, the SPMD and POCIS extracts were 

concentrated, in their original containers 

under a gentle stream of filtered nitrogen at 

ambient temperature, to approximately 0.250 

mL. After the extracts were concentrated, 

they were transferred to 1.8 ml gaschromatographic (GC) amber vials with 

Teflon lined screw caps, each containing a 

400 microliter (μL) insert. The volume of the 

extract was adjusted with hexane to 400 μL. 

Internal standards and injection internal 

standards were added to the extracts just prior 

to gas chromatographic and mass spectral 

(GC/MS) analysis. 

The extracts were analyzed by 

capillary GC/MS under electron ionization 

and electron-capture negative ionization. 

Table 2 lists the compounds that were 

analyzed for in the passive-sampler extracts. 

The conditions for the gas chromatographic 

and mass spectral analysis are described 

below. 

Capillary gas chromatographic analysis:

Chromatographic separations for all mass 

spectral analysis were made on an RTX-5MS 

30 M X 0.25 mm ID, 0.25 μm df fused silica 

capillary column. The injection port 

temperature was held at 285°C. The oven 

temperature program was as follows; the oven 

was held at 50°C for 5 min, the oven 

temperature was raised to 125°C at 10°C/min 

then raised to 200°C at 1.5°C/min, then raised 

up to 325°C at 8°C/min, and held at this 

temperature for 30 min. The transfer line 

temperature was held at 285°C. A 1-μL 

splitless injection of the concentrated extract 

was injected into the GC/MS for analysis. 

Ultrapure helium was the carrier gas. The 

carrier gas was filtered through moisture traps 

and traps for hydrocarbons and oxygen prior 

to entering the gas chromatograph. 

Mass spectral analysis:. For all mass

spectrometric analysis the instrument was 

repetitively scanned from 50 to 600 Daltons 

at a rate of 2.69 scans per second. The 

electron-multiplier was set at 2,000 volts. 

Application and specific conditions for each 

ionization mode are described below. 

First ionization: Electron-capture negative 

ionization (ECNI) mass spectrometry mode 

was used to measure pesticides, PCBs, 

polybrominated diphenyl ethers, and 

halogenated organic compounds in general in 

the extracts. The source temperature was 

held at 160o

C. The modifying gas was 

methane. The source pressure was 2 X 10-4

Torr. 

Second ionization: Electron ionization mass 

spectrometry is the conventional method for 

analyzing extracts via mass spectrometry. 

The source temperature was held at 200o

C. 

The source pressure was 4X 10-5 Torr. 

10 
Table 2. Chemical compounds analyzed for in passive-sampler (SPMD and POCIS) extracts and associated estimated 

detection levels. 

[ng/sampler, nanograms per sampler; ND, no detection; compounds detected in extracts from at least one site are italicized; 

SPMD, semipermeable membrane device; POCIS, polar organic compound integrative sampler]

Chemical 

compound 

Estimated 

detection 

level, in

ng/sampler 

Chemical 

compound 

Estimated

detection 

level, in

ng/sampler 

Trifluralin 4 o,p'-DDT ND

Benfluralin 4 cis-Nonachlor 128 

alpha-hexachlorohexane 16 Endrin Aldehyde 64 

Hexachlorobenzene 4 Endosulfan Sulfate 96

Pentachloroanisole 4 p,p'-DDT 32

beta-HCH ND Endrin Ketone 32

gamma-HCH 8 Mirex 4 

delta-HCH 32 PCB70 4

desulfinyl fipronil 16 PCB 101 4 

Aldrin 8 PCB 110 64

Chlorpyrifos 4 PCB 118 16

DCPA 4 PCB 138 4 

Octachlorostyrene 16 PCB 146 4

Heptachlor-epoxide 16 PCB 149 4 

Oxychlordane 16 PCB 151 4

fipronil sulfide 8 PCB 170 4 

fipronil 64 PCB 174 4 

trans-Chlordane 8 PCB 177 4 

o,p'-DDE 8 PCB 180 4 

Endosulfan I 8 PCB 183 4 

cis-Chlordane 96 PCB 187 4 

trans-Nonachlor 64 PCB 194 4 

p,p'-DDE 128 PCB 206 4 

Dieldrin 128 Toxaphene 2,500

o,p'-DDD 16 BDE 47 4 

Endrin ND BDE 99 4 

desulfinylfipronil amide ND BDE 100 4 

fipronil sulfone 8 BDE 153 4 

Endosulfan II 64 BDE 154 4 

p,p'-DDD 8

Compound identification: Identifications of 

the compounds detected in the extracts were 

classified as target or non-target (unknown). 

Target compounds were identified by 

comparing the mass spectra retention times 

for the sample extracts with the same 

properties from authentic standards, which 

are used to generate the standard curve for 

quantization. Unknown compounds were 

identified by comparing the mass spectrum of

the compound detected with a library of mass 

spectra, and if necessary, tentative 

identification was based on classical mass 

spectral interpretation. 

Quantitation: Quantization of the target 

compound was conducted using a 6-point 

calibration curve derived using linear 

regression, and authentic standards. Because 

authentic standards for the unknown 

compounds were not available, the 

quantization of an unknown compound is an 

estimate that is based on the response of the 

internal injection standard that is used for the 

quantitation of the target compounds. The 

quantization of the unknown compounds is a 

11 
semi-quantitative/estimated value. Although 

the reported concentration of the unknown 

compounds is an estimated value, the data 

may still be used to establish associations 

with toxicity tests. 

Blood Plasma 

Blood-plasma samples were extracted 

by solvent exchange, concentrated, and then 

analyzed for the compounds listed in table 3 

following procedures described in the 

preceding section, “Passive-Sampler 

Extracts.” 

Antibiotics 

Concentrations of antibiotics in water 

samples and POCIS extracts were determined 

by the USGS Organic Geochemistry 

Research Laboratory in Lawrence, Kansas. A 

total of 39 antibiotics in five antibiotic classes 

were analyzed for in both water samples and 

POCIS extracts (table 4). Samples were 

analyzed using online solid-phase extraction 

and Agilent 1100 Series liquid 

chromatography/mass spectrometers 

equipped with diode array detectors (Brown 

and others, 1999; Lindsey and others, 2001). 

Roxarsone and Other Arsenic Species 

Water concentrations of roxarsone, 

arsenate, arsenite, monomethyl arsonate, and 

dimethyl arsinate were determined by the 

USGS National Water Quality Laboratory 

(NWQL) in Lakewood Colorado, according 

to Garbarino and others (2002). Samples 

were filtered and preserved with EDTA, and 

then analyzed using inductively coupled 

plasma-mass spectrophotometry. 

Results 

The results of passive-sampler extract 

analysis, smallmouth bass blood-plasma 

analysis, stream-water analysis, and effluent 

analysis are presented in tables 5, 6, 7, and 8. 

Additionally, tables 2, 3, and 4 list all 

compounds except inorganic arsenic and 

organoarsenic compounds that were analyzed 

for in the various sample media, though not 

all compounds were detected. Compounds 

that were not detected in any environmental 

sample were omitted from the results tables. 

Passive-Sampler Results 

Both SPMDs and POCIS were 

deployed at six sites in the South Branch 

Potomac River Basin and one site each in the 

Cacapon River and Williams River Basins 

(table 1 and fig. 2). Table 2 lists 59 

compounds analyzed for in the passivesampler extracts; results of the analyses are 

shown in table 5. Four compounds were 

found at all sites; hexachloro-benzene (HCB), 

a fungicide banned from use in the United 

States; pentachloroanisole (PCA), a 

degradation product of other industrial 

phenols; 2,2',4,4'-tetra-bromo-diphenyl ether 

(BDE 47), a flame retardant; and 2,2',4,4',5-

penta-bromo-diphenyl ether (BDE 99), 

another flame retardant. Only these four 

compounds were detected in passive samplers 

deployed at Williams River at Dyer, the outof-basin reference site and the site with the 

fewest detected compounds. Other 

compounds detected in passive-sampler 

extracts are trifluralin, a dinitroanaline

herbicide; gamma hexa-chloro-hexane 

(γHCH), a pesticide; chlorpyrifos, an organophosphate pesticide; trans- and cis-chlordane, 

organo-chlorine pesticides; trans- and cisnonachlor, also organo-chlorine pesticides; 

and the flame retardant 2,2',4,4',6-pentabromo-diphenyl ether (BDE 100). Of the 

compounds detected the polybrominated 

diphenyl ethers (Meerts and others, 2001, 

Lilienthal and others 2006), trifluralin (Fox, 

2004), HCB (Ralph and others, 2003), γHCH 

(Kojima and others, 2004), chlorpyrifos 

(Kojima and others, 2004) cis-nonaclor (Fox, 

2004), and cis- and trans-chlordane (Kojima 

and others, 2004) are either known or 

12 
Table 3. Chemical compounds analyzed for in smallmouth bass blood-plasma samples and associated detection levels. 

[μg/kg, micrograms per kilogram; **, indicates compounds identified in reagent blanks, compounds identified in at 

least one plasma sample are italicized.] 

Chemical 

compound 

Detec

-tion

level, 

in

μg/kg 

Chemical 

compound 

Detec

-tion

level, 

in

μg/kg 

Chemical 

compound 

Detection 

level, 

in

μg/kg 

bromoform 25 AHTN (tonalide) 50 o,p'-DDD 32 

cumene 25 HHCB 50 p,p'-DDD 64

phenol 25 OPEO-1 100 o,p'-DDT 64

1,4-dichlorobenzene 25 carbaryl 250 p,p'-DDT 64

d-limonene 25 metalaxyl 250 dieldrin 16

acetophenone 25 anthraquinone 250 endosulfan I 8

para-cresol 25 bromacil 1,000 endosulfan II 8

isophorone 25 metolachlor 250 endrin 16

camphor 25 NPEO-1-total 1,500 endrin aldehyde 16

isoborneol 25 fluoranthene 25 endrin ketone 128

menthol 25 chlorpyrifos 100 endosulfan sulfate 4

napthalene 25 pyrene 25 fipronil 128

methyl salicylate 25 triclosan 250 alpha-HCH 32 

dichlorvos 25 OPEO-2 250 beta-HCH 64

isoquinoline 25 bisphenol A 250 gamma-HCH 8

indole 25 NPEO-2-total 2,000 delta-HCH 32

2-methylnapthalene 25 tri (dichloroisopropyl)

phosphate 

250 heptachlor-epoxide 16

2-methyl benzothiophene 100 triphenyl phosphate 250 hexachlorobenzene 2 

3,4-dichlorophenyl

isocyanate

100 ethanol, 2-butoxy-,

phosphate 

250 trans-nonachlor 8

1-methylnapthalene 25 diethylhexyl phthalate** 50 cis-nonachlor 4

skatol 25 beta-coprostanol 200 mirex 16

2,6-dimethylnapthalene 25 cholesterol 200 octachlorostyrene 2

butylated hydroxyanisole 25 beta-sitosterol 200 oxychlordane 16 

5-methyl-1Hbenzotriazole 800 stigmastanol 400 pentachloroanisole 2

diethyltoluamide (DEET) 25 aldrin 16 PCB70 64

diethyl phthalate** 25 benfluralin 2 PCB 101 32

4-tert-octylphenol 50 BDE 47 2 PCB 110 32

benzophenone 25 BDE 66 16 PCB 118 4

ethyl citrate 100 BDE 71 8 PCB 138 16 

tributylphosphate 500 BDE 85 4 PCB 146 2

cotinine 200 BDE 99 2 PCB 149 16

para-nonylphenol (total) 200 BDE 100 4 PCB 151 8 

pentachlorophenol 2,000 BDE 138 4 PCB 170 2

prometon 100 BDE 153 2 PCB 174 2

atrazine 50 BDE 154 4 PCB 177 2

phenanthrene 25 BDE 183 4 PCB 180 2

4-octylphenol 50 chlorpyrifos 8 PCB 183 2

tri (2-chloroethyl) 

phosphate 

100 trans-chlordane 2 PCB 187 2

anthracene 25 cis-chlordane 16 PCB 194 2

diazinon 100 DCPA 2 PCB 206 2

carbazole 25 o,p'-DDE 32 trifluralin 2

caffeine 50 p,p'-DDE 64 toxaphene 2,500

13 
Table 4. Antibiotics analyzed for in passive sampler extracts and in wastewater effluent in the South Branch Potomac

River, Cacapon River, and Williams River Basins. 

[*, degradation products; †

, detected in effluent discharge; μg/L, micrograms per liter] 

Analyte Laboratory reporting

level 

Analyte Laboratory reporting

level 

Beta lactams Sulfonamides

Amoxicillin 0.01μg/L Sulfachloropyridazine 0.005μg/L

Ampicillin 0.01μg/L Sulfadiazine 0.005μg/L

Cefotaxime 0.01μg/L Sulfadimethoxine 0.005μg/L

Claxacillin 0.01μg/L Sulfamerazine 0.005μg/L

Ocacillin 0.01μg/L Sulfamethazine 0.005μg/L

Penicillin G 0.01μg/L Sulfamethoxazole†

0.005μg/L

Penicillin V 0.01μg/L Sulfathiazole 0.005μg/L 

Macrolides and degradation products* Tetracyclines and degradation products*

Erythromycin

† 0.005μg/L Chlorotetracycline 0.01μg/L

Anhydro-erythromycin*†

0.005μg/L *Anhydro-chlorotetracycline 0.01μg/L

Lincomycin†

0.005μg/L *Demeclocycline 0.01μg/L

Ormetoprim 0.005μg/L Doxycycline 0.01μg/L

Roxithromycin 0.005μg/L Minocycline 0.01μg/L

Trimethoprim†

0.005μg/L Oxytetracycline 0.01μg/L

Tylosin† 0.005μg/L Tetracycline 0.01μg/L

Virginiamycin 0.005μg/L *Anhydro-tetracycline 0.01μg/L

Quinolines Other groups

Ciprofloxacin†

0.005μg/L Carbadox 0.005μg/L

Clinafloxacin 0.005μg/L

Flumequine 0.005μg/L

Lomefloxacin 0.005μg/L

Norfloxacin 0.005μg/L

Ofloxacin†

0.005μg/L

Oxolinic acid 0.005μg/L 

Sarafloxacin 0.005μg/L

14 
15 

suspected endocrine disruptors. The greatest 

number of compounds detected came from 

passive samplers deployed in the South 

Branch Potomac River at Petersburg Gap, 

with 12 compounds detected. POCIS extracts 

were also analyzed for the antibiotics listed in 

table 4. However, no detectable 

concentration of any antibiotic was identified 

in any POCIS extract. This finding was 

contrary to expectations, as samplers were 

deployed during spring flows when run-off 

events are common and following land 

application of poultry-house litter. 

Blood-Plasma Results 

A total of 30 smallmouth bass bloodplasma samples were collected from six sites 

(table 1 and fig. 2) and analyzed for the 

analytes shown in table 3. Ten of the analytes 

were present in detectable concentrations. 

Plasma samples were selected from both male 

and female mature fish without intersex 

characteristics and from male fish with 

intersex characteristics to represent the range 

of intersex conditions found at that site. 

Results of the analyses are shown in table 6. 

One compound, diethyl-hexyl phthalate, was 

found in every plasma sample analyzed. 

Diethyl-hexyl phthalate and diethyl phthalate 

were also detected in the reagent blanks. 

Reagent blank concentrations of diethyl-hexyl 

phthalate were well below concentrations 

found in plasma samples. However, reagent 

blank concentrations of diethyl phthalate (62 

mg/kg) approached those found in several 

plasma samples, therefore concentrations of 

diethyl phthalate in plasma maybe an artifact 

of reagent contamination. In addition to 

diethyl-hexyl phthalate and diethyl phthalate, 

the following compounds were detected in at 

least one plasma sample: phenol, an industrial 

compound with many applications; dlimonene, used as a solvent and also as a 

fragrance; acetophenone, used as a fragrance, 

a flavoring, and an industrial solvent; 

napthalene and 2-methyl-napthalene, used in 

polyvinyl chloride production and in both 

moth balls and toilet deodorizer blocks; 4-

tert-octyl-phenol, used as an industrial 

surfactant, plasticizer and in the synthesis of 

many end products; and the polybrominated 

diphenyl ether flame retardants 2,2',4,4'-tetrabromo-diphenyl ether (BDE 47) and 

2,2',4,4',5-penta-bromo-diphenyl ether (BDE 

99). Analysis of plasma samples from the 

Williams River reference site detected fewer 

compounds and those compounds were at 

lower concentrations than in the South 

Branch Potomac River, with the exception of 

phenol, which was found only in a plasma 

sample from the Williams River. The 

polybrominated diphenyl ethers, known 

endocrine disruptors, were found at all sites 

except the reference site. Diethyl-hexyl 

phthalate, a known endocrine disruptor, was 

found in every plasma sample. Patterns in 

diethyl phthalate, another known endocrine 

disruptor, are harder to discern due to 

possible reagent blank contamination. 

Water-Sample Results 

Water samples collected at the sites 

listed in table 1 and shown in fig. 2 were 

analyzed for concentrations of roxarsone and 

other arsenic species and antibiotics. The 

results of the analysis of seven samples of 

stream water and one effluent sample are 

shown in table 7. Arsenate, found in samples 

from six streams in the South Branch 

Potomac and Cacapon River Basins, in 

concentrations ranging from 0.3 μg/L to 1 

μg/L, was the only arsenic species detected. 

Arsenic concentrations in stream water 

samples from the South Branch Potomac 

River and Cacapon River Basins previously 

collected and analyzed by the USGS are 

typically low, with concentrations in 

dissolved-fraction samples ranging from 2 

μg/L to below detection in most samples. It 

was not possible to determine the source of 

the arsenate in the samples, whether from 

natural sources, coal combustion, or 

photodegradation of roxarsone. 
Table 5. Chemical compounds in passive samplers (SPMD and POCIS) deployed at sites in the South Branch Potomac River, Cacapon River, and Williams River 

Basins, April to June 2004. [ng/sampler, nanograms per sampler; <, less than; SPMD, semipermeable membrane device; POCIS, polar organic compound integrative sampler; RR, railroad] 

Map index number (fig. 1) 

Site name Trifluralin,

in ng/sampler

Hexachlorobenzene,in ng/sampler

Pentachloroanisole, in ng/sampler

gammaHexachlorohexane, in ng/sampler

Chlorpyrifos, in ng/sampler

transChlordane,in ng/sampler

cisChlordane,in ng/sampler

transNonachlor,in ng/sampler 

cisNonachlor,in ng/sampler 

2,2',4,4'-tetrabromodiphenyl ether (BDE 47), in ng/sampler

2,2',4,4',5-pentabromodiphenyl ether (BDE 99), in ng/sampler

2,2',4,4',6-pentabromodiphenyl ether (BDE 100), in ng/sampler 

1 So. Br. Potomac R 

near Upper Tract,WV 

< 4 3.4 13.6 33.4 < 4 11.7 12.3 12.2 < 4 16.8 14.4 6.74

2 N. F. South Br. 

Potomac R. at Cabins, WV 

< 4 7.7 14.9 34.5 20.1 8.14 9.4 9.4 < 4 14.9 11 < 4 

8 So. Br. Potomac. R.. 

at Petersburg Gap, WV 

75.4 5.34 12.3 21.3 15.1 9.94 11.4 10.6 4.8 13 9.6 5.5

9 South Branch

Potomac River near Moorefield, WV 

18.7 4.28 11.8 40.7 < 4 9.95 10.2 9.87 < 4 19.2 16.1 < 4 

13 So Br Potomac R at 

RR bridge at Sycamore, WV 

28.8 4.24 15 < 8 15.9 12.8 14.2 12.2 6.45 21.4 14 < 4

14 So. Br. Potomac R. 

near Springfield, WV 

10.1 7.45 15.4 43.6 < 4 12.5 15.1 12.2 < 4 27.4 19.4 8.21

15 Lost River at 

McCauley near Baker, WV 

< 4 2.88 25.3 < 8 70.8 6.51 <8 < 8 < 4 10.9 3.86 < 4 

16 Williams River at

Dyer, WV 

< 4 4.13 10.3 < 8 < 4 < 8 < 8 < 8 < 4 10.5 7.21 < 4 

16 
17 

Table 6. Chemical compounds in smallmouth bass blood-plasma samples from the South Branch Potomac River and the Williams River Basins, May to October 2004. [μg/kg, micrograms per kilogram; ng/mL, nanograms per milliter; <, less than] 

Map Index Number (fig. 1) 

Site name Fish 

sample ID 

phenol, in μg/kg 

d-limonene, in μg/kg 

acetophenone, in μg/kg 

napthalene, in μg/kg 

2-methylnapthalene, in μg/kg 

diethyl phthalate, in μg/kg 

4-tert-octylphenol, in μg/kg 

diethyl-hexyl phthalate, in μg/kg 

2,2',4,4'-tetra-bromodiphenyl ether (BDE 47), in ng/mL 

2,2',4,4',5-pentabromodiphenyl ether (BDE 99), in ng/mL 

SMB 1 <25 127 <25 41.5 47.5 <25 73 644 <2 <2 SMB 2 <25 152 47.1 <25 3.1 <2 

34 <25 <50 452 2.79 <2 

<25 41.9 94.3 261 

SMB 5 <25 <25 <25 41.8 44.5 <25 <50 335 8.4 2.79 SMB 7 <25 120 <25 25.6 42.7 <25 <50 638 <2 <2 

1 

South Branch Potomac River near Upper Tract, WV 

SMB10 <25 <25 <25 <25 

SMB 1 <25 177 95.4 <25 <25 108 <50 670 <2 4.11 

SMB 2 <25 74.2 62.8 <25 <25 66.8 <50 458 3.04 3.38 SMB 4 <25 85.5 <25 <25 <25 75.9 <50 339 13.13 2.64 SMB 6 <25 161 106 <25 <25 86.6 <50 884 <2 <2 SMB 7 <25 92.5 73.8 <25 <25 <25 <50 285 <2 <2 

SMB 12 <25 163 41.5 <25 <2 <2 

<25 101 <25 26.9 76 536 

1 <25 146 <25 29 30.6 <25 7.72 <2 

<25 29.1 97.8 715 

SMB 14 38 <25 4.2 <2 

2 

North Fork South Branch Potomac River at Cabins, WV 

SMB 16 <25 113 80.3 27.4 35.4 <25 <50 519 <2 <2 

SMB 71.2 376 SMB 4 <25 70.1 338 <2 

5 <25 <25 36.8 <25 3.49 <2 

<25 <25 <25 <25 <25 <2 

SMB <25 28.3 69.1 460 3 

South Branch Potomac River at Petersburg, WV SMB 6 <25 145 <25 33 50.9 <25 85.7 6,320 <2 <2 

SMB 5 <25 75.8 <25 <25 <25 78.8 <50 670 <2 <2 

SMB 7 <25 225 123 <25 <25 150 <50 304 <2 2.9 SMB 9 <25 141 <25 <25 <25 88.7 <50 366 2.5 <2 

SMB 10 <25 115 

<25 93 

78.8 <25 <25 103 <50 672 6.8 4.8 

8 

South Branch Potomac River at Petersburg Gap, WV 

SMB 15 63.4 <25 <25 74.2 2.9 <2 <50 2,070 

SMB 1 <25 134 86.5 <25 <25 <25 71.7 341 18.6 3.94 SMB 4 <25 135 <25 <25 <25 114 <50 216 26.3 5.23 SMB 9 69.4 <25 <25 <25 12.02 7.34 

15 <25 188 120 <25 <25 <25 131 1,080 7.43 5.4 

17 <25 <25 <25 128 <2 

<25 91.8 67.4 281 

SMB 

12 

South Branch Potomac River near Moorefield, WV 

SMB <25 90.9 82.1 378 <2 

SMB 2 38.9 32.8 55.8 <25 <25 94 <50 1,520 <2 <2 SMB 6 <25 69.8 103 <25 <25 110 <50 767 <2 <2 16 Williams River at Dyer, WV SMB 7 <25 150 <50 1,180 <2 <2 <25 <25 <25 <25 
18 

Samples from 4 selected effluent 

discharges were analyzed for the list of 39 

human and veterinary antibiotics and 

antibiotic degradation products in table 7, 

eight of which were detected in at least one 

sample (table 8). Only one antibiotic was 

detected in the sample from the slaughter 

house at Moorefield’s poultry processing 

plant discharge, ofloxacin, a human antibiotic 

not approved for veterinary use (U.S. Food 

and Drug administration, Center for 

Veterinary Medicine, FDA Approved Animal 

Drug Products (Green Book) web page, 

http://www.fda.gov/cvm/greenbook.html, 

accessed October 12, 2006) and its presence 

in the discharge was likely due to a human 

source. Fish hatcheries have been found to be 

sources of antibiotics (Dietze and others, 

2005); in this study both ofloxacin and 

anhydro-erythromycin were detected in the 

Spring Run hatchery discharge. The 

Petersburg sewage-treatment plant (STP) 

effluent sample contained detectable 

concentrations of seven antibiotics and one 

antibiotic degradate and the Moorefield STP 

had detections of six antibiotics and one 

antibiotic degradate. Concentrations of all 

antibiotics detected in the municipal STP 

effluents were above the median values in 

Glassmeyer and others (2005), except for 

ciprofloxacin, which was below the reporting 

level in the Moorefield STP effluent sample. 

Summary 

Passive water sampling devices used 

at eight stream sites detected the presence of 

emerging contaminants, including pesticide, 

flame-retardant, and personal-care product 

residues in water. No antibiotics or 

antibiotic-degradation products were found in 

passive sampler extracts from any site. 

Several of the compounds detected in passive 

sampler extracts are known or suspected as 

endocrine disrupting compounds (EDCs). 

These same compounds were also identified 

in blood plasma collected from smallmouth 

bass, including fish with intersex 

characteristics. Of the various arsenic species 

analyzed for, only arsenate was found in 

detectable concentrations. Antibiotics were 

found in the various wastewater effluents 

sampled, with municipal effluent having 

seven to eight compounds, whereas the fishhatchery and the poultry-processing plant 

effluents were found to have detections of 

two and one antibiotics, respectively. This 

study found that some EDCs are nearly 

ubiquitous in the environment, also being 

present at sites where fish are not exhibiting 

intersex conditions. Other investigations 

have shown that some EDCs, including some 

found in both passive samplers and 

smallmouth bass blood plasma, act additively 

or synergistically, where mixtures of several 

compounds have a greater effect than a single 

compound (Rajapakse and others, 2002; 

Payne and others, 2000). Studies are 

currently (2006) under way to further define 

the occurrence of EDCs in the South Branch 

of the Potomac basin and to determine the 

potential for environmental mixtures of these 

compounds to interfere with normal 

endocrine function. 
Table 7. Inorganic arsenic and organoarsenic compounds in stream-water and effluent samples collected at sites in the South Branch Potomac River and Cacapon 

River Basins, September 2004. [wf, water filtered using 0.45 μm capsule filter; μg/L, micrograms per liter; <, less than; SH, slaughter house] 

Mapindex number(fig. 1) 

Site name Sample date Sample time Arsenate, wf, in μg/L as As Arsenite, wf, in μg/L as As Dimethyl- arsinate, in μg/L as As Mono- methyl- arsonate, in μg/L as As Rox- arsone, in μg/L 

1 South Branch Potomac River near Upper Tract, WV 9/9/2004 1400 0.5 <0.3 <0.2 <0.2 <0.22 North Fork South Branch Potomac River at Cabins,

WV 

9/9/2004 1515 .4 <.3 <.2 <.2 <.2

5 Mill Creek near Petersburg, WV 9/9/2004 1600 .4 <.3 <.2 <.2 <.2 6 Lunice Creek at Petersburg, WV 9/9/2004 1630 <.3 <.3 <.2 <.2 <.2 9 South Fork South Branch Potomac River near 

Moorefield, WV

9/9/2004 1720 .3 <.3 <.2 <.2 <.2

11 SH Discharge At Moorefield, WV 9/10/2004 1020 1 <.3 <.2 <.2 <.2 13 South Branch Potomac River at railroad bridge at

Sycamore, WV 

9/10/2004 800 .4 <.3 <.2 <.2 <.2

15 Lost River at McCauley near Baker, WV 9/9/2004 1820 .4 <.3 <.2 <.2 <.2 Table 8. Antibiotics and antibiotic degradation products in composite samples collected from wastewater effluent in the South Branch Potomac River Basin,June 7-8, 2004.[μg/L, micrograms per liter; <, less than; SH, slaughter house; STP, sewage treatment plant; *, antibiotic degradation product] 

Macrolides and degradation products Quinolines Sulfonamides Map

index number(fig. 1) 

Site name 

Erythromycinin water, in μg/L 

Anhydroerythromycin*in water, in μg/L 

Lincomycinin water, inμg/L 

Trimethoprim in water, inμg/L 

Tylosin in water, inμg/L 

Ciprofloxacin in water, inμg/L 

Ofloxacin in water, inμg/L 

Sulfamethoxazole inwater, inμg/L 

4 Spring Run Hatchery <0.005 0.029 <0.005 <0.005 <0.005 <0.005 0.013 <0.005

7 Petersburg STP .442 1.47 .011 .031 .006 .053 .242 .12110 Moorefield STP .149 1.011 .005 .117 .007 <.005 .067 .21211 SH at Moorefield, 

WV

<.005 <.005 <.005 <.005 <.005 <.005 .009 <.005

19 
References Cited 

Alvarez, D.A., Petty, J.D., Huckins, J.N, 

Jones-Lepp, T.L., Getting, D.T., Goddard, 

J.P., and Manahan, S.E., 2004, 

Development of a passive, in situ, 

integrative sampler for hydrophilic organic 

contaminants in aquatic environments: 

Environmental Toxicology and Chemistry, 

v. 23, p. 1640-1648. 

Alvarez, D.A., Stackelberg, P.E., Petty, J.D., 

Huckins, J.N., Furlong, E.T., Zaugg, S.D., 

and Meyer, M.T., 2005, Comparison of a 

novel passive sampler to standard watercolumn sampling for organic contaminants 

associated with wastewater effluents 

entering a New Jersey stream: 

Chemosphere, v. 61, p. 610-622. 

Bevan, C. L., Porter, D. M., Prasad, Anita, 

Howard, M. J., and Henderson, L. P., 2003, 

Environmental estrogens alter early 

development in Xenopus laevis: 

Environmental Health Perspectives, v. 111, 

no. 4, p. 488-496. 

Brown, B. L., Slaughter, A. D., and 

Schreiber, M. E., 2005, Controls on 

roxarsone transport in agricultural 

watersheds: Applied Geochemistry, v. 20, 

no. 1, p. 123-133 

Brown, G. K., Zaugg, S. D., and Barber, L. 

B., 1999, Wastewater Analysis by Gas 

Chromatography/Mass Spectrometry, in 

Morganwalp, D.W. and H.T. Buxton, eds. 

U.S. Geological Survey Toxic Substances 

Hydrology Program—Proceedings of the 

technical meeting. Charleston, S.C., March 

8-12, 1999. Volume 2 of 3—Contamination 

of hydrologic systems and related 

ecosystems: U.S. Geological Survey-Water 

Resources Investigations Report 99-4018B. 

W. Trenton, NJ, 471 pp., 

http://toxics.usgs.gov/pubs/wri99-

4018/Volume2/index.html, p. 431-436 

Campagnolo, E.R., Johnson, K. R., Karpati, 

Adam, Rubin, C.S., Kolpin, D. W., Meyer, 

M. T., Esteban, J.E., Currier, R.W, Smith 

Kathleen, Thu, K.M., and McGeehin, 

Michael, 2002, Antimicrobial residues in 

animal waste and water resources proximal 

to large-scale swine and poultry feeding 

operations: The Science of the Total 

Environment, v. 299, no. 1-3, p. 89-95 

Carballa, Marta, Omil, Francisco, Lema, 

J.M., Llompart, Maria, Garcia-Jares,

Carmen, Rodriguez, Issac, Gomez, 

Mariano, Ternes, Thomas, 2004, Behavior 

of pharmaceuticals, cosmetics and 

hormones in a sewage treatment plant: 

Water Research, v. 38, p. 2918-2926 

Chapman, H.D. and Johnson, Z.B., 2002, Use 

of antibiotics and roxarsone in broiler 

chickens in the USA: Analysis for the years 

1995 to 2000: Poultry Science, v. 81, p. 

356-364

Dietze, J. E., Scribner, E. A., Meyer, M. T., 

and Kolpin, D. W., 2005, Occurrence of 

antibiotics in water from 13 fish hatcheries, 

2001-2003: International Journal of 

Environmental Analytical Chemistry, v. 85, 

no. 15, p. 1141-1152. 

Dyer, A. R., Raman, D. R., Mullen, M. D., 

Burns, R. T., Moody, L. B., Layton, A. C., 

and Saylrer, G. S., 2001, Determination of 

17β-estradiol concentrations in runoff from 

plots receiving dairy manure: American 

Society of Agricultural Engineers Meeting 

Paper no. 01-2107, Sacramento, Ca. 

Fox, Jennifer, 2004, Chemical 

communication threatened by endocrinedisrupting chemicals: Environmental 

Health Perspectives, v. 112, no. 6, pp 648-

653 

Garbarino, J.R., Bednar, A.J., and Burkhardt, 

M.R., 2002, Methods of analysis by the 

U.S. Geological Survey National Water 

Quality Laboratory---Arsenic speciation in 

natural-water samples using laboratory and 

field methods: U.S. Geological Survey 

Water-Resources Investigations Report 02-

4144, 40 p. 

Glassmeyer, S. T., Furlong, E. T., Kolpin, D. 

W., Cahill, J. D., Zaugg, S. D., Werner, S. 

L., Meyer, M. T., and Kryak, D. D., 2005, 

Transport of chemical and microbial 

compounds from known wastewater 

discharges- Potential use as indicators of 

20 
human fecal contamination: Environmental 

Science and Technology, v. 39, p. 5157-

5169. 

Goodbred, S. L., Gilliom, R. J., Gross, T. S., 

Denslow, N. P., Bryant, W. L., and Schoeb, 

T. R., 1997, Reconnaissance of 17βestradiol, 11-ketotestosterone, vitellogenin, 

and gonad histopathology in common carp 

of United States streams: Potential for 

contaminant-induced endocrine disruption: 

U. S. Geological Survey Open-File Report 

96-627, Sacramento, CA, 47 p. 

Hanselman, T. A., Graetz, D. A., and Wilkie, 

A. C., 2003, Manure-borne estrogens as 

potential environmental contaminants: A 

review- Environmental Science and 

Technology, v. 37, no. 24, p. 5471-5478 

Hutchison, M.L., Walters, L. D., Avery, S. 

M., Muro, F. and Moore, A., 2005, 

Analyses of livestock production, waste 

storage, and pathogen levels and prevalence 

in farm manures: Applied and 

Environmental Microbiology, v. 71, no. 3, 

pp. 1221-1236 

Huckins, J., Petty, J., Prest, H., Clark, R., 

Alvarez, D., Orazio, C., Lebo, J., Cranor, 

W., Johnson, B., 2002, A guide for the use 

of semipermeable membrane devices 

(SPMDs) as samplers of waterborne 

hydrophobic organic contaminants.: API 

Publication 4690. American Petroleum 

Institute, Washington, DC

Jobling, S, Beresford, N, Nolan, M., RodgersGray, R., Tyler, C.R. and Sumpter, J.P., 

2002, Altered sexual maturation and 

gamete production in wild roach (Rutilus 

rutilus) living in rivers that receive treated 

sewage effluents: Biology of Reproduction. 

V 66, no. 2, p. 272-281. 

Kavanagh, R. J., Balch, G. C., Kiparissis, 

Niimi, A. J., Sherry, Jim, Tinson, Cheryl, 

and Metcalfe, C. D., 2004, Endocrine 

disruption and altered gonadal development 

in white perch (Morone americana) from 

the lower Great Lakes Region: 

Environmental Health Perspectives, v. 112, 

no. 8, p. 898-902. 

Kojima, Hiroyuki, Katsura, Eiji, Takeuchi, 

Shinji, Niiyama, Kazuhito, and Kobayashi, 

Kunihiko, 2004, Screening for estrogen and 

androgen receptor activities in 200 

pesticides in In Vitro reporter gene assays 

using Chinese hamster ovary cells: 

Environmental Health Perspectives, v. 112, 

no. 5, p. 524-531 

Kolodziej, E. P., Harter, Thomas, and Sedlak, 

D. L., 2004, Dairy wastewater, aquaculture 

and spawning fish as sources of steroid 

hormones in the aquatic environment: 

Environmental Science and Technology, v. 

38, no. 23, p. 6377-6384 

Kolpin, D. W., Furlong, E. T., Meyer, M. T., 

Thurman, E. M., Zaugg, S. D., Barber, L. 

B., and Buxton, H. T., 2002, 

Pharmaceuticals, hormones, and other 

organic wastewater contaminants in U. S. 

streams, 1999-2000- A national 

reconnasissance: Environmental Science 

and Technology, v. 36, no. 6, p. 1202-1211 

Lilienthal, Helmuth, Hack, Alfons, RothHärer, Astrid, Grande, S. W., and Talsness, 

C. E., 2006, Effects of developmental 

exposure to 2,2’,4,4’,5-

pentabromodiphenyl ether (PBDE-99) on 

sex steroids, sexual development, and 

sexually dimorphic behaviour in rats: 

Environmental Health Perspectives, v. 114, 

no. 2, p.194-201 

Meerts, I. A. T. M., Letcher, R. J., Hoving, 

Saske, Marsh, Göran, Bergman, Åke, 

Lemmen, J. G., van der Burg, and Brouwer, 

Abraham, 2001, In vitro estrogenicity of 

polybrominated diphenyl ethers, 

hydroxylated PBDEs, and polybrominated 

bisphenol A compounds: Environmental 

Health Perspectives, v. 109, no. 4, p.399-

407 

Melo, A. C., and Ransdell, J. S., 2001, Sexual 

dimorphism of brain aromatase activity in 

Medaka- induction of a female phenotype 

by estradiol: Environmental Health 

Perspectives, v. 109, no. 3, p. 257-264. 

Nash, J. P., Kime, D. E., Van der Ven, L. T. 

M., Wester, P. W., Brion, François, Maack, 

Gerd, Stahlschmidt-Allner, Petra, and 

21 
Tyler, C. R., 2004, Long-term exposure to 

environmental concentrations of the 

pharmaceutical ethynylestradiol causes 

reproductive failure in fish: Environmental 

Health Perspectives, v. 112, no. 17, p. 

1725-1733. 

Nichols, D. J., Daniel, T. C., Moore, P.A. Jr., 

Edwards, D. R., and Pote, D. H., 1997, 

Runoff of Estrogen hormone 17β-estradiol 

from poultry litter applied to pasture: 

Journal of Environmental Quality, v. 26, 

no. 4, pp 1002-1006. 

Orlando, E. F., Kolok, A. S., Binzcik, G. A., 

Gate, J. L., Horton, M. K., Lambright, C. 

S., Gray, L. E. Jr., Soto, A. M., and 

Guilette, L. J. Jr., 2004, Endocrinedisrupting effects of cattle feedlot effluent 

on an aquatic sentinel species, the fathead 

minnow: Environmental Health 

Perspectives, v. 112, no. 3, p. 353-358. 

Paybins, K.S., Messinger, Terence, Eychaner, 

J.H., Chambers, D.B., and Kozar, M.D., 

2000, Water Quality in the Kanawha–New 

River Basin West Virginia, Virginia, and 

North Carolina, 1996–98: U.S. Geological 

Survey Circular 1204, 32 p., on-line at 

http://pubs.water.usgs.gov/circ1204/ 

Payne, Joachim, Rajapakse, Nissanka, 

Wilkins, Megan, and Kortenkamp, 

Andreas, 2000, Prediction and assessment 

of the effects of mixtures of four 

xenoestrogens: Environmental Health 

Perspectives, v. 108, no. 10, p. 983-987. 

Petty, J.D., Huckins, J.N., Alvarez, D. A., 

Brumbaugh, W. G., Cranor, W.L., Gale, R. 

W., Rastall, A. C., Jones-Lepp, T.L., 

Leiker, T.J., Rostad, C.E., and Furlong, 

E.T., 2004, A holistic passive integrative 

sampling approach for assessing the 

presence and potential impacts of

waterborne environmental contaminants: 

Chemosphere, v. 54, no. 6, p. 695-705 

Rajapakse Nissanka., Silva, Elisabete, and 

Kortenkamp Andreas, 2002, Combining 

xenoestrogens at levels below individual 

no-observed-effect concentrations 

dramatically enhances steroid hormone 

action: Environmental Health Perspectives 

V. 110, no. 9, 917-921. 

Ralph, J. L., Orgebin-Crist, Marie-Claire, 

Lareyre, Jean-Jacques, and Nelson, C. C., 

2003, Disruption of androgen regulation in 

the prostate by the environmental 

contaminant hexachlorobenzene: 

Environmental Health Perspectives, v. 111, 

no. 4, p. 461-466 

Raman, D. R., Williams, E. L., Layton, A. C., 

Burns, R. T., Easter, J. P., Daugherty, A. S., 

Mullen, M. D., and Salyer, G. S., 2004, 

Estrogen content of dairy and swine wastes: 

Environmental Science and Technology, v. 

38, no. 13, p. 3567-3573 

Scheurs, R. H. M. M., Legler, Juliette, ArtolaGarciano, Elsa, Sinnige, T. L., Lanser, P. 

H., Seinen, Willem, and Van ser Burg, 

Bart, 2004, In vitro and in vivo

antiestrogenic effects of polycyclic musks 

in zebrafish: Environmental Science and 

Technology, v. 38, no. 4, pp 997-1002. 

Schmitt, C.J., T.M. Bartish, V.S. Blazer, T.S. 

Gross, D.E. Tillitt, W.L. Bryant, and L.R. 

DeWeese, 1999, Biomonitoring of 

Environmental Status and Trends (BEST) 

Program: Contaminants and their effects in 

fish from the Mississippi, Columbia, and 

Rio Grande basins. Pp. 437-446 in 

Morganwalp, D.W. and H.T. Buxton, eds. 

U.S. Geological Survey Toxic Substances 

Hydrology Program—Proceedings of the 

technical meeting. Charleston, S.C., March 

8-12, 1999. Volume 2 of 3—Contamination 

of hydrologic systems and related 

ecosystems: U.S. Geological Survey-Water 

Resources Investigations Report 99-4018B, 

W. Trenton, NJ, 471 pp. 

http://toxics.usgs.gov/pubs/wri99-

4018/Volume2/sectionD/2507_Schmitt/inde

x.html

Schreiber, M.E., Chambers, D.B., and White, 

J. S., 2004, Cycling of organoarsenic 

compounds in agricultural watersheds 

[abs]: Eos, Transactions, American 

Geophysical Union, v. 85, no. 47, Fall 

Meeting Supplement, Abstract H21C-1031. 

22 
Sheahan, D.A., Brighty, G.C., Daniel M, 

Jobling, S., Harries, JE., Hurst, M.R., 

Kennedy J., Kirby, S.J., Morris, S., 

Routledge, E. J., Sumpter, J.P., Waldock, 

M.J., 2002, Reduction in the estrogenic 

activity of a treated sewage effluent 

discharge to an English river as a result of a 

decrease in the concentration of industrially 

derived surfactants: Environmental 

Toxicology and Chemistry, v. 21, no. 3, p. 

515-519. 

Toft, Gunnar, Edwards, T. M., Baatrup, Erik, 

and Guillette, L. J. Jr., 2003, Disturbed 

sexual characteristics in male mosquitofish 

(Gambusia holbrooki) from a lake 

contaminated with endocrine disruptors: 

Environmental Health Perspectives, v. 111, 

no. 5, p. 695-701. 

U.S. Census Bureau, 2002, Summary File 1, 

census of population and housing: 

http://www.census.gov/PressRelease/www/2001/sumfile1.html 

U.S. Department of Agriculture, National 

Agricultural Statistics Service, 2004, West 

Virginia state and county data: 2002 census 

of agriculture, v. 1, Geographic Area Series 

part 48, 513 p. 

(http://www.nass.usda.gov/census/census02

/volume1/wv/WVVolume104.pdf) 

U.S. Environmental Protection Agency, 2004, 

2002 Toxic Release inventory data release: 

http://www.epa.gov/tri/tridata/tri02/index.h

tm

U. S. Food and Drug Administration, 2000. 

Freedom of Information Summary NADA 

140-445: U.S. Food and Drug 

Administration, 

http://www/fda/gov/cvm/infores/foi/140445

062888.html

West Virginia Department of Environmental 

Protection, 2006, 2006 integrated water 

quality monitoring and assessment report: 

West Virginia Department of 

Environmental Protection, Charleston, WV, 

53 p. 

23