MOBILE RIVER STUDY

1993-1994

10103035

ENVIRONMENTAL SERVICES DIVISION

ECOLOGICAL SUPPORT BRANCH

960 COLLEGE STATION ROAD

ATHENS, GEORGIA 30605-2700
Table of Contents

Summary, Conclusions, and Recommendations i

L . O Introduction 1

1.1 Goal 1

2.0 Study Approach 2

2.1 Study Implementation 2

2.2 Study Area 3

3.0 Screening Assessment 4

3.1 Sampling Strategy and Methods 4

3.2 River and Floodplain Sampling Results 5

3.2.1 In Situ Measurements 5

3.2.2 River Water 6

3.2.3 River Sediments 6

3.3 Floodplain Sampling Results 7

3.3.1 Surface Water 7

3.3.2 Sediments 9

4.0 Fish Tissue Contamination Assessment 10

4.1 Sampling Strategy and Methods 10

4.2 Sampling Results 11

4.2.1 Fish Fillets 11

4.2.2 Whole Body Fish 15

4.2.3 Floodplain Fish 18

5.0 Fish Health Index Assessment 19

5.1 Sampling Strategy and Methods 19

5.2 Autopsy Results 20

6.0 Discussion and Review of Results 21

6.1 Public Health Considerations 25

6.2 Preliminary Ecological Risk Evaluation 26

6.2.1 Assumptions 26

6.2.2 Exposure Pathways 27

6.2.3 Exposure Profiles 28

6.2.3.1 Wood Stork 28

6.2.3.2 Belted Kingfisher 28

6.2.4 Effects Profiles 29

6.2.4.1 Mercury 29

6 . 2.. 4 . 2 DDTr 30

6.2.5 Risk Characterization . 32

6.2.6 Sources of Uncertainty 32

6.2.7 Results and Conclusions 33

7.0 References 35

Tables

Figures

Appendices
Tables

1. Potential Chemicals of Concern Associated with

Superfund Sites.

2. Sequence of Sampling Efforts for the Mobile

River Study.

3. Minimum Detection Limit Ranges for Potential

Chemicals of Concern for All Fish Tissue

Samples Analyzed.

4. Organic Compounds Detected in River Surface

WdCer Samples. October 1993.

5. Results of Inorganic Analyses of River Surface

Water Samples. October 1993.

6. Results of Inorganic Analyses of River Sediment

Samples. October 1993.

7. Particle Size and Organic Carbon Content as

Percent Dry Weight of River Sediments (October, 1993)

and Floodplain Sediments (April, 1994).

8. Mobile River Study, August 7, 1993.

9. Results of Organic Analyses of S'.rface Water Samples

from the Floodplain Wetlands. October 1993.

10. Results of Inorganic Analyses of Surface Water

Samples from the Floodplain Wetlands. April 1994.

11. Results of Inorganic Analyses of Sediment from the

Floodplain Wetlands. April 1994.

12. Results of Organic Analyses of Floodplain Sediments.

April 1994.

13. Results of Organic Analyses of Largemouth Bass Fillet

"Composite Samples. November 1993.

14. Results of Organic Analyses of Channel Catfish

Composite Fillet Samples. November 1993.

15. Mean and Range of Lipid Content for Largemouth Bass

and Channel Catfish fillet and Whole Body Samples.

1993 - 94.

16. Results of Inorganic Analyses of Largemouth Bass Fillet

Samples. November 1993.

17. Results of Inorganic Analyses of Channel Catfish Fillet

Samples. Nover^er 1993.

18. Results of Organic Analyses of Largemouth Bass Whole

Body Samples. November 1993.

19. Results of Organic Analyses of Channel Catfish Whole

Body Samples. November 1993.

20. Mean and Range of Detectable Concentrations of DDE, ODD

and DDT in Fillet and Whole Body Samples of Largemouth

Bass and Channel Catfish. November 1993.

21. Results of Inorganic Analyses of Largemouth Bass Whole

Body Samples. November 1993.

22. Results of Inorganic Analyses of Channel Catfish Whole

Body Samples. November 1993.

23. Results of Inorganic Analyses of Whole Body Forage Fish

Composite Samples. November 1993.

24. Results of Organic Analyses of Whole Body Forage Fish

Composite Samples. November 1993.
25. Results of Inorganic Analyses of Whole Body Samples of

Fish Collected from the Floodplain Wetlands. April

1994 .

26. Results of Organic Analyses of Whole Body Fish from the

Floodplain Wetlands. April 1994.

27. Fish Health Index Scores for Largemouth Bass. October

1994.

28. Fish Health Index Scores for Channel Catfish. October

1994.

29. Mean and Range of TOTAL DDT concentrations in Fillet

and Whole Body Samples of Largemouth Bass and Channel

Catfish. April 1994.

30. Mean and Range of Mercury Concentrations in Fillet

Samples. November 1993.

31. Mean and Range of Mercury Concentrations in Whole Body

Samples. November 1993.

32. Summary of Human Risk Levels from Appendix G.

33. Hazard Quotient Calculations with Supporting

Documentation for the Wood Stork (Mycteria americana)

ingesting Mercury-contaminated prey within the 53-mile

Mobile River Study Area, 1993-94.

34. Hazard Quotient Calculations with Supporting

Documentation for the Wood Stork (Mycteria americana)

ingesting DDT-contaminated prey within the 53-mile

Mobile River Study Area, 1993-94.

35. Hazard Quotient Calculations with Supporting

Documentation for the Belted Kingfisher (Ceryle alcyon)

ingesting Mercury-contaminated prey within the 53-mile

Mobile River Study Area, 1993-94.

36. Hazard Quotient Calculations with Supporting

Documentation for the Belted Kingfisher (Ceryle alcyon)

ingesting DDT- contaminated prey within the 53-mile

Mobile River Study Area, 1993-94.
Figures

1. Mobile/Tombigbee Rivers Sampling Stations and Areas.

2. Site Locations for Water Quality and Sediment Sampling

in River and Floodplain Wetlands, Station MT-0

October, 1993 and April, 1994, respectively.

3. Site Location for Water Quality and Sediment Sampling

in the Mobile River, Station MT-1, October 1993,

respectively.

4. Site Location for Surface Water and Sediment Sampling

the river station MT-2 and in the Floodplain Wetlands

associated with Cold Creek Swamp. October, 1993 and

April, 1994 respectively.

5. Site Locations for Water Quality and Sediment Sampling

in the River (MT-7), and Floodplain Wetlands MT-7S, and

MT-7N, October 1993, and April 1994, respectively.

6. Site Location for Water Quality and Sediment Sampling

in the River MT-9, and MT-8, and Floodplain Wetlands,

MT-8, October 1993 and April 1994, respectively.

7. Site Locations for Water Quality and Sediment Sampling

in the Floodplain Wetlands, MT-11, April 1994. Tensaw

Lake area of Tensaw River.

8. Sampling Site Locations and Results for Total Mercury,

and Temperature in the Floodplain Wetlands associated

with Cold Creek Swamp, and Station MT-2, April 1994.

9. Sampling Sites and Results for Total Mercury in Canal

Surface Water, November, 1994. Barry Steam Electric

Generating Facilities, Cooling Water return to the

Mobile River.

10. Length of Age Frequency for Largemouth Bass from the

Upper Mobile Delta, Mobile River. Fall, 1991.

11. Fillet Samples Exceeding DDE Criteria Relative to

Station, Species and Size. November 1993.

12. Fillet Samples Exceeding ODD Criteria Relative to

Station, Species and Size. November 1993.

13. Fillet Samples Exceeding DDT Criteria Relative to

Station, Species and Size. November 1993.

14. Relationship Between Channel Catfish Length and Fillet

Concentration of Total DDT. November 1993.

15. Relationship Between Largemouth Bass Length and Fillet

Concentration of Total DDT. November 1993.

16. Relationship Between Size of Channel Catfish and Fillet

Concentration of Mercury. November 1993.

17. Relationship Between Size of Largemouth Bass and Fillet

Concentration of Mercury. November 1993.

18. Range and Mean Concentration of Mercury in Largemouth

Bass Fillet Samples Prepared of Large Fish. November

1993.

19. Range and Mean Concentration of Mercury in Channel

Catfish Fillet Samples. Nrv Briber 1993.

20. Relationship Between Larc^ruoith 3c.ss Total Length and

Whole Body Concentration of fetal OD"1. November 1993.
21. Relationship Between Channel Catfish Total Length and

Whole Body Concentration of Total DDT. November 1993.

22. Relationship Between Whole Body Lipid Content and Size

of Largemouth Bass. November 1993.

23. Relationship Between Whole Body Lipid Content and Size

of Channel Catfish. November 1993.

24. Average and Range,of Total DDT Concentration of

Largemouth Bass Whole Body Samples. November 1993.

25. Average and Range of Total DDT Concentration of Channel

Catfish Whole Body Samples. November 1993.

26. Relationship Between Size of Largemouth Bass and Whole

Body Concentration of Mercury. November 1993.

27. Relationship Between Size of Channel Catfish and Whole

Body Concentration of Mercury. November 1993.

28. Average and Range of Mercury Concentrations in

Largemouth Bass Whole Body Samples Relative to Sampling

Station. November 1993.

29. Average and Range of Mercury Concentrations in Channel

Catfish Whole Body Samples Relative to Sampling

Station. 1993-1994.

30. Average and Range of Total DDT Concentration of Forage

Fish Whole Body Samples. November 1993.

31. Mean Fish Health Index (FHI) Scores for Largemouth Bass

and Channel Catfish. November 1993.

32. Cumulative Distribution Frequency for Largemouth Bass

Fillet Concentration of DDE. November 1993.

33. Cumulative Distribution Frequency for Channel Catfish

Fillet Concentration of DDE. November 1993.

34. Temporal Trend in Total DDT Concentration of Largemouth

Bass Whole Body and Fillet Tissue Samples from the

Mclntosh Area of Tombigbee River.''

35. Temporal Trend in Total Mercury Concentration of

Largemouth Bass Whole Body and Fillet Tissue Samples

from the Mclntosh area of the Tombigbee River.
Appendices

A. Plan of Study (POS) and Related Amendment.

B. In Situ Measurements and Results and Mean and Monthly

River Flows.

C. Fish Collection Protocol, Mobile River Study, 1993-

94.

D. Fish Health Index (FHI) Protocol.

E. Autopsy Field Data Sheets.

F. Spreadsheets Summarizing of Autopsy Data and Calculated

FHI Scores.

G. Human Health Risk Evaluation of Fish Data, Mobile River

Study.

H. Responsiveness Summary.
Summary, Conclusions, and Recommendations

Summary

• The goal of the Mobile River Study was to assess the

contamination of the river, including those hazardous substances

and contaminants associated with and potentially emanating from

four Superfund National Priority Listed Sites and entering the

Mobile-Tombigbee River system. To assess the potential effects

of these contaminants, a 53 mile reach of the river system was

designated for study. This reach extended upstream from River

Mile Marker 19 near the Interstate 1-65 bridge to a point

cipproximately 13 miles upstream of Mclntosh, Alabama.

Study objectives included the characterization of the surface

v/ater and sediments of the river and associated floodplain

wetlands, assessment of the chemical contamination of Largemouth

Bass, Channel Catf-ish and forage fish, and determination of the

health condition of the bass and catfish.

The field investigation activities spanned a period extending

c- om September, ]993 through early Novemlet:,

 n994. During this

period, six data collection surveys were conducted. Of this

number, five were conducted in the river and one in the

floodplain wetlands associated with the river while under a

period of inundation by river flow.

• Of the myriad of organic compounds and metals analyzed for in

sediments and water, mercury and the pesticide DDT and related

metabolites DDE an ODD emerged as the chemical constituents of

potential concern in the study reach of the river. The

analytical detection of DDT and metabolites was limited mainly to

sediments of the river and floodplain wetlands intimately

associated with the Olin Basin. Total mercury concentrations

exceeding EPA criteria for surface water were restricted to

samples collected from the Cold Creek Swamp area when flooded by

river water.

• With the exception of cyanide and volatile organic compounds,

fish tissues were analyzed for the same chemical constituents as

were the sediments and surface water. One hundred sixty-two

fillet composite samples of Largemouth Bass and Channel Catfish

were prepared from small, medium and large size groups and

analyzed. About 120 whole body samples of bass, catfish and

forage fish were chemically analyzed. As was the 'case for the

sediments and water, total mercury and the pesticide DDT and

related metabolites DDE and ODD emerged from the analyses as the

only chemical constituents found to be a potential concern.
• Relative to public health considerations, mercury levels in

the fillet samples of Largemouth Bass and Channel Catfish were

well below the US Food and Drug Administration (USFDA) action

level and EPA criteria of one mg/kg. A limited human health

assessment yielded Hazard Quotients (HQs) for mercury between 0.1

and 1.0. These values were within EPA's acceptable level of an

HQ less than 1.

Total DDT concentrations of the Largemouth Bass and Channel

Catfish fillet samples were well below the action level of 5

mg/kg established by the USFDA. However, DDE, ODD, And DDT

concentrations exceeded the State of Alabama adopted criteria in

10 and 7 percent of the bass and catfish fillet samples analyzed,

respectively. Twenty-six and 53 percent of the bass and catfish

fillet samples contained concentrations of these compounds that

exceeded EPA criteria. A limited human health risk assessment

.yielded an increased risk (between 1:10,000 and 1:100,000) of

cancer under the subsistence fisherman scenario. This scenario

assumes a subsistence fisherman eating 6 ounces of contaminated

fish from an area of the river adjacent to the Olin Basin and

Ciba-Geigy floodplain, twice a week, for j,0 years.

• A preliminary ecological risk evaluation was conducted using

exposure scenarios involving a Wood Stork and a Belted Kingfisher

as representative target receptors for the study area. The

evaluation demonstrated a potential for adverse ecological

effects, particularly for fish eating birds. The belted

kingfisher and the wood stork were chosen as representative

receptors for the exposure assessment.

• Results of the Fish Health Index indicate that Largemouth Bass

and Channel Catfish were in relatively good condition with the

exception of encysted parasitic worms in the spleens, livers, and

kidneys which was common to all stations including the reference

and background stations. Lipid content of the fish were within

the range of values associated with background and reference

data.

• The mean concentration of mercury was significantly greater in

the fillets of Channel Catfish collected from the river station

position adjacent to the Cold Creek Swamp which is associated

with drainage from the Stauffer NPL Sites.

• Concentrations of total DDT were detected in nearly all fish

samples. The highest mean concentrations of total DDT in fillet

and whole body samples of catfish, bass, and forage fish were

found in the river area associated the Mclntosh sampling station

and the Olin and Ciba-Geigy NPL Sites. Historical data coupled

with results of this study suggest a decline in the contamination

level of total DDT in the Largemouth Bass and probably also in

other fish.

ii
Conclusions

» The findings of this study demonstrate that sediments of the

river and floodplain wetlands that are in close association with

the Olin and Ciba-Geigy NPL sites (MT-7), are contaminated with

total DDT and mercury. Fish from the river and floodplain

wetlands in close association with these same- sites, were

contaminated with DDT and related metabolites. In addition,

elevated concentrations of total mercury occurred in surface

water and sediments of the floodplain wetland and in fish from

the river area associated with the Stauffer LeMoyne and Cold

Creek Plant NPL Sites (MT-2).

• Overall, the limited human health risk assessment indicates

that the risk to the general public was marginal or within EPA

criteria; however, a potential increased risk to a subsistence

fisherman was demonstrated.

• The preliminary ecological risk evaluation indicates that, in

general, there is no potential effect to large, fish eating,

wading birds as represented by the woodstork. A potential for

ecological effects is evident for the belted kingfisher and those

fish eating birds having similar ecological functions and feeding

regimes.

These results indicate that the four NPL Sites and their

associated wetlands are potential sources to the river system.

Recommendations

• Given the conclusions stated above, this report recommends

remedial action for the floodplain wetlands of the four NPL

Sites. In addition, active monitoring of the river should be

implemented in the vicinity of the Cold Creek Swamp in Axis and

the Olin Basin/Ciba-Geigy floodplain in Mclntosh. In addition, a

continued monitoring program for the river system should be

developed to determine that contaminant levels in the river

channel continue to decrease, as indicated in the finds of this

report, to more acceptable levels.

111
1.0 Introduction

The Mobile/Tombigbee River is a waterway extending north

from Mobile Bay to Pickwick Reservoir on the Tennessee River.

Along its course upstream from Mobile Bay are four Superfund

National Priority List (NPL) Sites. The Stauffer Chemical

{LeMoyne Plant) Site located in Axis, Alabama and the Stauffer

Chemical (Cold Creek Plant) Site located in Bucks, Alabama are

approximately 20 miles north of Mobile, Alabama, at River Mile

26. The Olin Corporation (Mclntosh Plant) and the Ciba-Geigy

Corporation (Mclntosh Plant) are located in Mclntosh, Alabama,

approximately 50 miles north of Mobile, Alabama at River Mile 60

(Figure 1).

Each of these Sites has backwater or floodplain wetland

areas that are adjacent to and are hydrologically connected to

the Mobile/Tombigbee river system. From review of background

information and results of on-going investigations initiated

through the CERCLA process, it has been shown that each of these

wetland areas contain sediments containing elevated levels of

IIP --ardous substances. This information sui /ey provided a basis

for the development of a list of Chemicals of Concern (Table 1).

The presence of these contaminants in the wetland areas and the

knowledge that the wetland areas are intimately related to the

Mobile/Tombigbee river system, lead the U.S. Environmental

Protection Agency (EPA) to suspect that the River system had been

contaminated by the activities of these four NPL Sites. In

addition, other study findings reported by the U.S. Fish and

Wildlife Service (USFWS), Alabama Department of Environmental

Management (ADEM), and EPA have shown elevated levels of mercury

and DDT in the tissue of fish captured from the river segment and

in the vicinity of the CERCLA sites. Mercury is a potentially

toxic constituent known to be associated with the Stauffer and

Olin NPL sites. The compound DDT and its metabolites DDE and

ODD are also Chemicals of Concern associated with the Olin and

Ciba-Geigy NPL Sites.

EPA believes that the demonstrated potential releases of

contamination may pose a threat to human health and the

environment. Therefore, the EPA, Region IV, Waste Management

Division, Office of Superfund, initiated the Mobile River Study

to address the Agency's concern.

1.1 Goal

The goal of the Mobile River Study is to assess the

contamination of the river, including those hazardous substances,

pollutants and contaminants associated with and potentially

emanating from each or all of the NPL Sites and entering the

designated river segment. To facilitate this goal, the following

objectives were established:

• Characterization of the quality of water and sediments

of the river segment and. • -sociated wetland areas in
terms of potential chemicals of concern,

• Assessment of the chemical contamination of selected

and representative fish species or communities of the

river segment,

Determination of the health condition of a selected top

predator game fish species of the river segment.

The data collected from the Mobile River Study will be used

to better characterize the contamination in the river segment,

assess the potential risks to human health and the environment,

and assess contributions of contamination from the four NPL

Sites.

2.0 Study Approach

2.1 Study Implementation

The technical approach to this study vras described in a Plan

of Study (POS). Preceding implementation of ^ne POS, the

document was subject to technical and program review by the EPA,

the four companies associated with the NPL Sites, the federal and

state Natural Resource Trustees, and the Public. With some

exceptions as later discussed, the study proceeded as indicated

in the POS. An amendment to the POS was implemented in the later

stages to reflect some special sampling considerations during

high water level conditions in selected floodplain wetland areas

of the river. The POS and amendment are provided in Appendix A.

In brief, the POS describes study rationale, study design

considerations including quality assurance and data quality

objectives, sampling design and methods, and general data

analysis. The implementation of the POS is next reported

including deviations and additions, study observations and

results, data discussions and summary and conclusion.

The principle activities of the study were conducted in two

phases. The first phase, or screening assessment segment of the

study, involved sampling at strategically selected stations to

determine the general presence, relative distribution and

concentration of constituents for characterizing the quality of

surface water and sediments of the study reach. From this

effort, the results were used to determine the scope of sampling,

if required, to complete the chemical characterization of the

water and sediments in a following effort of sampling.

The sampling framework for the second phase of study was

organized to provide a basis for comparisons between station

parameters, with a primary focus upon assessing fish tissue

contamination and the fish health condition of Largemouth Bass

and Channel Catfish. The combined sampling activities for the
complete study spanned the period of time extending from mid

September, 1993 through early November, 1994. The sequence of

sample activities are provided in Table 2.

2.2 Study Area

The study as implemented involved a reach of the

Mobile/Tombigbee River system extending approximately 53 miles

upstream from River Mile 20 marker located near the Interstate I65 bridge. Seven stations were sampled for analysis of surface

water and sec'iment. Nine stations were sampled for fish

analysis, as indicated in Figure 1. Stations MT-8 and MT-9 were

used as background sites. Two additional sites were also

selected for reference purposes with one located on the Alabama

River (MT-10) near Chrysler, Alabama and the second site on the

Tensaw River in the Tensaw Lake region (MT-11). The former site

was chosen because of its long term use in the USFWS's National

Fish Contamination Study, while the latter was selected for

purposes of providing a reference location for characterizing

chemicals of concern in the river associated floodplain wetlands.

The tudy reach was fairly typical of the Mobile/Tombigbee River

system in terms of general near shore condition and habitat,

river width, and depth. Although the river's navigation channel

was maintained at a minimum depth of nine feet, depths exceeding

35 feet were frequently encountered over the course of the study

reach.

River currents were most pronounced in the maintained

channel even at stages of low flow. Common to the near shore

areas of the river were eddies created by a meandering shoreline

characterized by land 'points and shore pockets which appeared to

be mostly created by bank erosion. The bank erosion appeared

more severe and common along the shoreline of the Tombigbee River

section of the study reach. Tidal amplitude effects of Mobile

Bay were only readily apparent in the vicinity of stations MT-0

and MT-2 which were located at the most downstream segment of the

study area.

From a reconnaissance of the study reach in September, 1993,

the principle types of substrate characterizing the navigational

channel of the Tombigbee section of the study were primarily a

mixture of coarse to medium sand, small gravel, and shell

fragments. This prominent substrate feature reflected the

effects of strong, scouring currents and to some degree, possibly

barge traffic. The near shore substrate of the Tombigbee River

featured mainly fine to medium sand and traces of silt.

For the Mobile River section of the study area, beginning at

the confluence of the Alabama River, sediment composition was

typically represented by a substrate of medium to coarse sand in

the navigation channel. The bottom areas flanking the channel

and the shoreline were generally characterized by a sediment
mixture of fine sand and silt.

Since the Mobile/Tombigbee waterway is maintained for

navigation and not flood control, overbanking of the river during

storm flow conditions occurs. With overbanking, nutrients and

settleable solids are supplied to the river floodplain wetlands.

These organic and inorganic materials could include the potential

chemicals of concern. For these reasons, selected floodplain

areas were included in the sampling strategies.

To help characterize flow conditions of the river monthly

mean flow records for the study period were compared to long

terms records in Figures B-l and B-2 (Appendix B). The flow

information was provided by the U.S. Geological Survey for the

Coffeeville and Claiborne Lock and Dam locations. These records

reflect runoff from about 93 percent of the Mobile River drainage

basin.

3.0 Screening Assessment

3.1 Sampling Strategy and Methods

To narrow the array of potential chemicals of concern as

listed in Table 1, the screening phase of the study involved two

sampling periods. In early October, 1993, the river was sampled

during low flow stages. The floodplain wetlands associated with

the river were later sampled under conditions of inundation in

early April, 1994. The flooded condition provided access to the

sampling areas by boat.

A selected number of sediment and water samples from the

study reach were collected and analytically screened for the

chemical constituents. The stations sampled were MT-0

(downstream of 1-65 bridge), MT-1 (upstream approximately one

mile from 1-65 bridge), MT-2 (downstream of Cold Creek swamp),

MT-7 (downstream of Olin Basin at Mclntosh), MT-8 (Three Rivers

Lake, a backwater area approximately four miles upstream of

Mclntosh), MT-9 (approximately five miles upstream of Mclntosh),

and MT-11 in the Tensaw Lake area of the Tensaw River near

Brant's Landing, Alabama.

The sampling scheme at each of the screening stations on the

river was initiated by first determining the vertical

distribution of temperature, dissolved oxygen (DO), conductivity,

and pH in the water column using field calibrated electronic

probes. These profiles help to determine the presence and degree

of physical and chemical stratification of the water column and

the depth at which to obtain the most representative samples of

the river's water column. Sampling of the water column was

accomplished with a DC electrically operated peristaltic pump

which served to evacuate a precleaned four liter sample bottle.
From the bottle, an intake line of precleaned teflon tubing was

extended to the water depth selected for sampling. The four

liter collection bottle when full was used to pour into separate

sample bottles for analyses. Samples for volatile analyses were

taken directly from the teflon intake line when used as a

capillary tube.

Water sampling of the inundated floodplain wetlands was

accomplished from a boat. Individual sample bottles were

directly submerged to a depth of approximately one foot and

filled. For collection of water samples for ultra trace level

analysis of mercury, the person collecting the sample wore a

second pair of plastic gloves which extended the length of the

arm. For each floodplain station, multiple sites were sampled

for surface water and sediment analyses (Figures 2, 3, 4, 5, 6

and 7). The sampling sites in the floodplain wetlands are

bracketed for clarity. These sites were haphazardly selected and

not georeferenced with GPS readings; thus, their position shown

in the former figures were general approximations relative to the

river and other land marks. During the course of sampling in the

floodplain wetlands, samples of fish were also collected for

tissue analyses. Because of the active sampling strategy of

electrofishing, the fish samples represented a composite of

specimens rather than discreet samples for a specific site as was

the case for water and sediments. Results of the fish tissue

analyses are reported later in the fisheries section.

Sediment samples were collected from the river's navigation

channel and the shallower, off channel areas of the river bed.

Initially, sampling was accomplished at the first station (MT-0)

with diver operated hand cores, this was subsequently

discontinued, for the efficiency and safety of a petite ponar

dredge deployed from the boat. In the floodplain wetlands,

sampling was accomplished with a hand held auger which was fitted

with an extension handle, which was operated from a sampling

boat. By either means of sampling, multiple samples of sediments

from each site were composited to ensure a sufficient sample size

was obtained for chemical and particle size analyses.

3.2 River and Floodplain Sampling Results

3.2.1 In Situ Measurements

The results of the in situ measurements of DO, temperature,

conductivity and pH are detailed in Appendix B. These data

indicated the water column of the river was well mixed vertically

with the exception of the navigation channel at station MT-0 and

MT-2. The depth at which water sampling was conducted are

indicated in Table B-l. Density stratification in conjunction

with safety concerns associated with prolonged anchoring in the

navigator's channel were principal considerations in choosing

sampling locations. The type sampling device employed for the
collection of water samples required sustained anchoring for a

period exceeding one hour. Barge traffic during the anchoring

period proved to be a significant safety concern.

3.2.2 River Water

Although the water samples were analyzed for a great number

of organic compounds, very few contaminants were found at

concentrations exceeding minimum detection limits (MDL) of the

selected analytical procedures (Table 3). This table provides

the range of minimum detection limits per parameter as

encountered during the course of their respective analyses. The

variation mainly reflected differences in individual sample

variables such as moisture content, sample size of extraction

aliquot, and matrix composition. Subsequently, the MDL varied

greatly between some of the samples which were analyzed for the

same constituent.

The results of the organic analyses identified only three

comoounds (Table 4). Atrazine was detected at low concentrations

ir samples from station MT-7 near Mclntosh ^nd Stations MT-8 and

MT-9 which were both background locations located about four to

five miles upstream from Mclntosh. Atrazine is listed as a

chemical of concern associated with the Ciba-Geigy NPL site.

EPTC, a thiocarbamate compound, was detected in low

concentrations at station MT-2, MT-0 and MT-1, near the 1-65

bridge.

The results of the inorganic analyses indicated an array of

metals present in all water samples (Table 5) . Of this array,

copper and mercury were the only constituents listed among the

chemicals of Concern. Mercury was found at very low

concentrations at all stations with values ranging from 1.7 to

4.9 ng/1. Copper concentrations ranged from less than 2.0 ug/1

to 7.4 ug/1. Other than copper, the concentrations of the metals

detected were generally comparable to or less than what was

observed at the two background stations MT-8 and MT-9.

3.2.3 River Sediments

The objective of sediment sampling in the screening study

was to characterize the chemical quality of sediments and

determine if the chemical constituents and concentrations varied

according to sources such as the NPL Sites and particle size of

the bottom substrate. An assumption involved in this sampling

approach was that sorptive processes coupled with settling

properties of inorganic and organic particles would favor the

accumulation of chemicals in sediments characterized by finer

substrate particles.

The sediment analyses resulted in the quantification of 15

metals and no detectable concentrations of organic compounds. Of
the quantified metals, copper and lead were the only chemicals of

concern detected. Their concentrations, as well as the other

metals reported, were subject to a wide degree of variation

between samples (Table 6).'

The sediment samples were subjected to a particle size

analyses which yielded a suite of size fractions reported as

percent composition by dry weight (Table 7). Two principal types

of bottom substrates were determined from the analyses. Sampling

sites designated as MT-OC, MT-2A1, A2, A3, and MT-8 were

characterized by a substrate comprised primarily of fine sand and

silt. Sediments from sites MT-OB and MT-1C were characterized by

a predominance of coarse' to medium sand and traces of silt. A

characterization also applied to station MT-9 as determined by a

field inspection rather than a laboratory analysis. A sediment

sample of station MT-9 was returned to the laboratory for

•analyses but lost in the process. Given these physical

differences in sediment, those substrates sharing like particle

size composition also appear to share similar metal

concentrations (Table 6).

Two months prior to the above sampling, the personnel of the

Center for Applied Isotope Studies of the University of Georgia

(UGA) conducted a survey of bottom sediments for trace and heavy

metal concentrations (Noakes 1993). Their sampling effort

extended from River Mile 5 on the Mobile River and upstream to RM

78 on the Tombigbee River. Within this reach, box core grab

samples of sediments were collected from 22 locations along the

center line of the river. Each sediment sample was sieved and

the particle fraction passing through a 200 mesh screen (75

micron mesh opening) was analyzed for metals. Their findings are

summarized in Table 8. Many of the elements reported in the UGA

study such as Cadmium and Mercury were not detected in the

screening study of EPA. One aspect of the UGA study was that the

metal analysis involved exclusively very fine sediments because

of the sieving process in preparing the samples for analysis.

The results of the UGA study indicate that the metals

concentrations of the river sediments both upstream and

downstream of the NPL Sites were similar, as the EPA survey.

3.3 Floodplain Sampling Results

3.3.1 Floodplain Surface Water

Of the organic compounds analyzed in water samples from the

floodplain survey, Atrazine and Simazine were the only compounds

found at detectable concentrations (Table 9). Both compounds are

labeled as selective herbicides with the former being in wide use

and Simazine to a lesser extent. These compounds also feature

relatively low solubility in water and are common constituents of

farm field runoff (Grover 1989). With the exception of station

MT-0, Simazine, a listed chemical of concern associated with the

7
Ciba-Geigy NPL site, was detailed in at least one of the three

sites sampled at each station. Atrazine was common to all

stations. Concentrations of these herbicides ranged from 0.020

to 0.55 ug/1 and 0.037 to 0.06 ug/1, respectively. The wide

distribution of these two herbicides across the study area

including the background and reference stations suggests

watershed sources of these compounds other than or in addition to

the Ciba-Geigy NPL site.

The inorganic analyses of the floodplain water samples yield

an array of t^race and heavy metals (Table 10) . Among these

metals reported, copper, lead, and mercury were the only elements

identified as potential chemicals of concern. Mercury was common

to all the surface water samples. Except for station

MT-2, total mercury concentrations ranged from 2.8 to 9.6 ng/1.

The exception involved the three floodplain sites of station

'MT-2 in the lower floodplain wetland area of Cold Creek Swamp

(Figure 4). Water samples from these three sites contained total

mercury concentrations ranging from 37 to 63 ng/1. These values

exceeded EPA criteria for surface water and background and

reft • ance sites by an order-of-magnitude. J.ie detection of

copper in surface waters was limited to the three sampling sites

of the floodplain of the lower Cold Creek Swamp MT-2(A), 2(B),

and 2(C) and one background site at station MT-8(C).

Concentrations of copper were 6.0, 6.6, 7.0, and 3.2 ug/1 for

these sites, respectively.

Another aspect of the survey findings from the inundated

floodplain of the lower Cold Creek Swamp were elevated surface

water temperatures. At the time of the sampling, surface water

temperatures of 22.0, 22.1, and 26.0 C were measured at sites

MT-2(A), 2(B), and 2(C), respectively. Floodplain surface water

temperatures measured elsewhere at stations MT-0, 7, 8, and 11

ranged from 16.8 to 18.0 C. The elevated water temperature

appeared attributable to the circulation of heated water into the

swamp from the nearby Barry steam electric generating facility.

At the time-of these observations, overbanking of the discharge

canal was evident. This overbanking could explain the apparent

circulation pattern.

Assuming there is circulation of heated effluent to the Cold

Creek Swamp, this effluent did not appear to be as a factor

contributing to the elevated values of mercury in the swamp. The

temperature gradient coupled with that of the mercury

concentration observed in the swamp suggests the heated effluent

may be having a diluting effect on the mercury regime of the

swamp (Figure 8). Sampling of the steam electric facility's

intake and discharge canal were later sampled for mercury

analyses. Results of these analyses indicated that the waste

water discharges from the facility was not an apparent source of

the mercury (Figure 9). Samples for these analyses were

collected in November, 1994 during a river stage when flows were
contained within the banks of the canal. The concentration of

these samples ranged from, 2.5 to 3.9 ng/1 and-were comparable to

background concentrations reported for the October, 1993, survey

of the river. These results would suggest that the elevated

mercury concentrations in Cold Creek Swamp were not likely

derived from the waste water discharge of the Barry steam

electric generating facility but rather from drainage originating

in the swamp.

3.3.2 Floodplain Sediments

Organic analyses of sediments from the river's floodplain

wetlands indicated the presence of few quantifiable compounds

(Table 12). The compound DDT and related metabolites were found

mainly in the sediments of the floodplain wetlands between the

Olin Basin and the river (Figure 5). Of the three sampling

'sites, MT-7C(A), MT-7S(B), and MT-7S(C), the latter station was

positioned about 40 feet southwest of the vegetational edge of

the floodplain and actually in the river bed. Sediment

concentrations of total DDT at these stations were 338, 95, and

SO mg/kg, respectively. The latter two concentration were an

order-of-magnitude greater than the ER-L value of 3 mg/kg which

was established by Long and Morgan, 1990 as a concentration above

which adverse effects to sensitive species of aquatic may be

initiated. For station MT-7S(A), the sediment concentrations of

338 mg/kg of total DDT approached the ER-M value which was

considered to be a concentrations above which adverse effects are

almost always observed or predicted.

Concentrations of trace and heavy metals from MT-7S(C) were

substantially less when compared to the other floodplain sites

but similar to the river's bottom substrate characterized by fine

sediments at river sites MT-OC and MT-2A, 2B, and 2C (Table 6).

The sediments from site MT-7S(C) were also characterized by the

predominance of fine sand and silt (Table 7).

The other floodplain sediments were also in sharp contrast

to MT-7S(C) as well as the other river sediments. The silt-clay

fractions of the floodplain sediments were two to three times

greater than observed at MT-7S(C) and elsewhere in the river.

The greater content of silt and clay of the floodplain sediments

probably helps explain the greater concentration regimes of

metals which were found in the floodplain substrates of the

river.

The analytical results for the floodplain sampling sites

indicate similar inorganic chemical constituents and

concentrations among the stations with the exception of mercury

at sites MT-O(C), MT-2(B), MT-2(C) and MT-7S(A) (Table 11). At

these sites, sediments sconcentrations of mercury ranged from 3.3

to 32 mg/kg. The concentrations were one to two orders-ofmagnitude greater in concentrations compared to other sites and
exceed the ER-M value of 1.3 mg/kg. The EM-L value is a sediment

concentration above which adverse effects can be expected or

predicted (Long and Morgan, 1990) . Mercury is a chemical of

concern identified for both the Stauffer and Olin NPL Sites which

was associated with the latter three sampling sites,

respectively. Site MT-O(C) was located downstream of the

Stauffer Site.

4.0 Fish Tissue Contamination Assessment

4.1 Sampling Strategy and Methods

The focus of the fishery work was to assess the chemical

contamination of Largemouth Bass, Channel Catfish and forage

fish. The study approach t.o this objective involved

electrofishing from a boat and the use of cheese baited slat box

traps measuring 15 X 15 X 72 inches. The fish collection efforts

for the river and inundated floodplain wetlands occurred at

different times of the year. For the river sampling, fish

collection efforts were in November and December, 1993. Fish

collection in the floodplain wetlands occurred in April, 1994.

To assess chemical contamination of the targeted species,

fillet and whole body samples were analyzed for most of the

chemicals of concern listed in Table 1. Cyanide and volatile

organic compounds were excluded from these analyses. The fish

sampling and processing protocols leading to these analyses

involved the collection of sufficient numbers of the desired

species of different size groups.

For the Largemouth Bass, length at age frequency data for

the upper Mobile River Delta area were obtained from the Alabama

Division of Game and Fish, Fishery District 5 at Spanish Fort,

Alabama (Figure 10). From these data, three size groups were

selected for the Largemouth Bass which were: 170 to 225 mm

(small), 226 to 500 mm (medium), and 301 to 400 mm (large). In

each size group, the length of the smallest fish size was

targeted to equal to or greater than 75 percent of the length of

the largest fish of the group. In most cases, the targeted size

range was obtained. To develop the three size groups for Channel

Catfish, samples of the species were collected from the upper and

lower reaches of the study area. From these collections, a size

distribution for the species was obtained and the following

length determined: 200 to 275 mm (small), 276 to 375 mm (medium),

and 376 to 500 mm (large). Total length was the standard unit of

length measurement for both bass and catfish. The protocol for

processing the fish after capture is provided in Appendix C.

Each size group was represented by three replicate composite

samples of fillets for inorganic analyses and three replicate

composite samples of fillets for organic analyses. To accomplish

10
the whole body analyses, several specimens of the two predator

species were collected from each station. For each species, an

array of sizes were selected. The forage fish samples were

comprised of an assemblage of small species which usually

included menhaden, silversides, small sunfish, shad and minnows.

Six replicate composite samples of forage fish were collected

from each station and divided between the organic and inorganic

analyses.

As expected the catch per unit of effort for the species of

interest varied among the study stations. For this reason, the

river area for the collection of fish also varied among stations

because of the fixed sample size need. The general fishing areas

are depicted in Figure 1. These same fishing areas also served

for the Fish Health Index as later reported.

A special sampling effort was extended to the floodplain

wetlands when they were inundated during a high river flow period

in April, 1994. The purpose of the special sampling was to gain

some understanding of the presence and chemical condition of the

targeted species inhabiting the wetlands during the inundation

period. The sampling strategy was.to collect adequate numbers of

fish to constitute a sample of sufficient fillet and whole body

biomass to represent the three size groups of the targeted

species. The stomach contents of the captured fish were examined

to determine the types of food items consumed;these contents were

analyzed for the chemicals of concern. The limited fish sampling

was accomplished with electrofishing gear.

4.2 Sampling Results

4.2.1 Fish Fillets

Approximately 820 specimens of Largemouth Bass and Channel

Catfish, were collected and processed for laboratory analyses.

The identification and quantification of several organic

compounds resulted (Tables 13 and 14). Of these compounds DDT

and related metabolites DDE and ODD were the only observed'

organic chemicals of concern. Other pesticides reported were

detected in only a few samples and were widely dispersed among

the sampling stations including background and reference sites.

As indicated in Tables 13 and 14, composite samples of

fillets representing stations MT-0, MT-2, MT-3, and MT-7

underwent partial thawing in shipment to the laboratory for

analyses. Seventeen duplicates of these samples were later

shipped and received in the frozen condition by the same

laboratory and analyzed. With one exception, which was a bass

fillet composite from station MT-0, the duplicate results were in

good agreement with the original analyses. The difference in the

total DDT concentration between the thawed and frozen duplicates

were within the range of variance expected in split samples for
an inter-laboratory analyses of fish tissue for total DDT (EPA,

1993a,1995a). Subsequently, the initially reported results for

the partially thawed samples were treated as uncompromised

samples.

The pesticide DDT and its related metabolites were listed as

chemicals of concern because of their association with the Olin

and Ciba-Geigy NPL sites at Mclntosh, Alabama (Table 1). The

metabolites DDE and ODD are typically a result of natural

degradation processes of DDT. Subsequently, the presence of DDE

or ODD in fish tissue at concentrations greater than DDT are

common since DDT was banned from manufacturing in 1972. Such is

the case for the results of the fillet analyses.

The metabolite DDE was found at detectable concentrations in

94 percent of all samples for both Largemouth Bass and Channel

Catfish with ODD occurring in 28 percent and DDT in 6 percent of

the total fillet samples. Several of theses samples yielded

concentrations exceeding Alabama's recently adopted criteria and

EPA criteria for human consumption (USEPA 1993). Concentrations

of D})E in the fillet ? of the bass ranged f re - the MDL to 0.27"

mg/kg with 21 and six percent of the values equal to or greater

than EPA's and Alabama's criteria of 0.0316 and 0.069 mg/kg,

respectively. The concentrations for DDD ranged from less than

detection to 0.350 mg/kg with three observation exceeding the

EPA criteria of 0.0449 mg/kg and two exceeding the Alabama

criteria of 0.096. The measured concentrations for DDT ranged

from 0.007 to 0.130 mg/kg and were limited to four composite

samples from station MT-7 at Mclntosh. Only one of these samples

exceeded the EPA and Alabama criteria of 0.0316 and 0.069 mg/kg,

respectively.

In Channel Catfish fillet composite samples, concentrations

of DDE ranged from 0.005 to 0.290 mg/kg with 53 and seven percent

of the values exceeding the EPA and Alabama criteria which are

cited above. For DDD, the catfish fillet samples ranged in

detectable concentrations from 0.005 'to 0.091 mg/kg with five

percent of the samples exceeding the EPA criteria. Detectable

levels of DDT were limited to five samples from station MT-7 at

Mclntosh where concentrations ranged from 0.005 to 0.048 mg/kg.

Two of these samples exceeded the EPA criteria.

With respect to station location, species, and size group,

the distributions of those samples exceeding EPA criteria for

DDE, DDD, DDT are shown in Figures 11, 12, and 13. The

bioaccumulation of these pesticides to levels exceeding EPA

criteria was most prominent in Channel Catfish. The incidence of

excessive concentrations was mainly represented in the large and

medium size catfish. The large bass were the size class

appearing most effected. No detectable concentrations of any of

the pesticides were found in fillet samples from Station MT-10 on

the Alabama River.

12
The distribution of samples exceeding EPA criteria among the

stations downstream of MT-7 was fairly uniform. Excluding MT-7,

the proportion of samples•exceeding the DDE criteria at each of

the Mobile/Tombigbee stations ranged from 22 to 44 percent with

the extreme of this range being represented by reference station

MT-8. The number of possible samples for any one station was 18.

At station MT-7, 72 percent of the fillet samples exceed the EPA

criteria for DDE. For DDD and DDT, samples exceeding the EPA

criteria were fewer in number but exclusive to station MT-7

(Figures 12 and 13).

To determine if total DDT concentrations were significantly

different between stations, the data were treated in the

following manner. A data set was first examined for normality of

distribution and homogeneity of variance. Where necessary, the

data sets were transformed and tested to assure a normal

distribution and homogeneity of variance. A model I ANOVA was

followed by an unplanned multiple comparison procedure which

employed Duncans multiple range test to discern differences

between stations. This particular test was chosen because of its

po''t.: to detect differences. Total DDT vala^s In the data sets

were determined by adding together the reported concentrations of

DDE, DDD, and DDT for each of the samples analyzed. In many

cases, the respective compounds were reported at a concentration

defined as minimum detection limit (MDL). The MDL observations

were included in the total DDT calculations by assuming that the

median of the MDL as a concentration representing the compound.

Using the Duncans Multiple Range Test at the Alpha 0.05

level, a comparison of mean fillet concentrations between station

was conducted for total DDT concentrations in the fish samples.

For catfish fillet composite samples, the mean concentration of

total DDT for station MT-7 was significantly greater than all

other stations. The mean concentration of total DDT in bass

fillets was also significantly greater than other stations except

in the comparison with the reference station MT-8 where nosignificant differences in mean concentrations were detected.

Another observation that can be derived from Figures 11, 12,

and 13 is that the larger fish reflected a greater proportion of

the samples exceeding criteria with catfish appearing to be more

prone to the accumulation of these pesticides in the fillet

tissue than the bass. Since DDT and its metabolites are

lipophilic soluble, bioaccumulation would be more favored in

those fish and species with the greater body lipid content. This

appears to be the case with the Channel Catfish and Largemouth

Bass. The present lipid content of the catfish fillets appeared

greater when compared to the bass (Table 15).

To examine the relationship between fish size and pesticide

concentration, average length of the five fish comprising each

fillet composite and the total DDT concentration of the composite

13
sample were compared through linear regression (Figures 14 and

15). Total DDT was determined as the sum of detectable DDT, DDE,

and DDD concentrations for each fillet composite. The median of

the reported MDL were included in the determination. As shown in

these two figures, the fillet concentration of total DDT was

weakly correlated with fish length. The R SQUARE values for the

Channel Catfish and Largemouth Bass regression lines were 0.143

and 0.090, respectively. Of the concentration array plotted for

total DDT, no concentration approached the value of 5 mg/kg which

is the action level designated for total DDT by the USFDA as

protective of human health.

Several trace and heavy metals were detected in the

inorganic analyses of fillet samples. For both species, about

half of these elements were represented by one or at most a few

observations scattered among the stations (Tables 16 and 17). Of

these infrequently occurring elements, copper and lead were

included.

The array of metals of common detection among the fillet

samples included calcium, magnesium, manganese, nickel,

potassium, sodium, zinc, and mercury. In most instances, the

range in the concentrations per constituent remained similar to

values observed for the background and reference stations. Since

mercury was identified as a particular metal of concern, its

distribution relative to species, size of fish and station are

considered in the following.

Through linear regression analyses, the relationship between

the total length of fish and fillet concentration was considered

for Channel Catfish and Largemouth Bass (Figures 16 and 17). As

shown in the former figure, the relationship between fish length

and mercury concentration in the catfish was weakly developed as

indicated by a low R Square value of 0.067. The regression

analyses indicates that on the average, mercury concentrations in

the catfish fillet samples remained fairly independent of the

fish size. The relationship between fish length and mercuryconcentration in the Largemouth Bass fillet samples was much more

developed as indicated by a R Square value of 0.653. In this

analyses, the mercury concentration of the bass fillets appear

much more dependent on size of fish than the catfish.

With a more focused view on size effects, the mean and range

of concentrations of fillets prepared from the large size groups

of Largemouth Bass were plotted relative to station number

(Figure 18). For the large size bass, the average concentration

of mercury in the fillet samples were bounded by a large degree

of variation about the average including the background stations.

Stations MT-2, MT-3, MT-4, MT-8, MT-9 and MT-10 appeared similar

in range and concentrations.

Mercury concentrations reported for the Channel Catfish

14
fillets appeared less variable about the mean than with the bass

(Figure 19). With exception of station MT-2, the average

concentration of mercury for each station varied from 0.05 to

0.11 mg/kg'. At station MT-2, mercury ranged from 0.06 to 0.54

mg/kg and averaged 0.23 mg/kg which was nearly twice the value

for other stations. The maximum mercury concentration in the

reference sites ranged from 0.04 to 0.16 mg/kg. Based upon the

observed distribution of this metal among the sampling sites,

mercury contamination of the Channel Catfish was most evident at

station MT-2, a site in close association with Cold Creek Swamp

and the Stauffer NPL site. A Duncan's multiple range test at

alpha 0.05 shows the mean concentration of fillets for station

MT-2 was significantly greater compared to all other stations.

The reported mercury concentrations for the catfish and bass

fillet samples were all substantially less than 1 mg/kg which is

the action level of the USFDA and EPA criteria for human

consumption (USFDA AND EPA 1995).

4.2.2 Whole Body Fish

The organic analyses of the whole body samples of Channel

Catfish and Largemouth Bass resulted in the quantification of

five organic compounds which were Endosulfan I (ALPHA),

Chlorobenzilate, DDT, DDE, and DDD (Tables 18 and 19). The last

four compounds were identified as chemicals of concern.

Endosulfan I was limited to a single Largemouth Bass sample from

station MT-10, a reference site on the Alabama River.

Chlorobenzilate was detected in three of five whole body catfish

samples found at station MT-3. This same pesticide was detected

in a whole body sample of bass from each of the five different

stations including a reference and background site. Collectively

among the two species, the detectable concentrations of

Chlorobenzilate ranged from 0.041 to 0.083 mg/kg with 0.054 and

0.065 mg/kg representing the background and reference stations

(MT-9 and MT-10).

Principal pesticides of concern in this study were DDT and

its related metabolites DDE and DDD. Of the total number of

whole body analyses of Largemouth Bass (49 fish) and Channel

Catfish (42 fish), 96 and 100 percent, respectively, contained

detectable concentrations of DDE. For DDD, 35 and 42 percent of

the respective samples contained this compound with DDT being

limited to 12 percent of the whole body samples. As expected,

the DDE levels in both species represented maximum concentrations

relative to DDD and DDT. Regarding the latter two compounds,

little differences occurred between the fillet and whole body

concentrations per species.

The relationship between fish size and concentration of

total DDT was examined through linear regression analysis. The

median values for the reported MDL observations were included in

15
the calculation of total DDT concentrations. The correlation of

fish length with total DDT concentration was considered through

linear regression analyses for both bass and catfish (Figures 20

and 21). Both regression analyses yielded relatively low R

SQUARE values of 0.17 and 0.18, respectively. These values

suggest that total DDT concentration for whole body were weakly

dependent of fish size. Over all, the whole body concentrations

of total DDT for the Largemouth Bass averaged about 30 percent

greater than the average reported for Channel Catfish (Table 20).

This difference between the two species; however, does not appear

to be a function of whole body percent lipid content as might be

expected (Table 15). Percent lipid content of both whole body

and fillets were, with few exceptions, greater in catfish than in

the bass. Increased concentration of total DDT, however, favored

the bass in terms of whole body contamination. The range of

percent lipid content for both species were comparable to those

values reported in National Contamination Bioaccumulation Program

of the USFWS (Schmitt, Zajicek, and Perterman,1990). Recent

lipid content of whole body bass correlated moderately well with

fish size as indicated by a R-SQUARE of 0.38 (Figure 22). For

whole body catfish, the correlation between size and percent

lipid concentration was poor as indicated by a R-SQUARE of 0.08

(Figure 23).

The distributions of total DDT concentrations in terms of

means and ranges for whole body samples of the bass and catfish

relative to sampling station are depicted in Figures 24 and 25.

The median of the of the reported MDL were include in the

determination of total DDT. Using Duncan's multiple range test

at an alpha of 0.05, the mean concentration of total DDT in whole

body Largemouth Bass was not significantly different between any

station sampled except for the reference site (MT-10).

Similarly, mean whole body concentrations of total DDT in catfish

were not significantly different between stations except for

sampling sites MT-8 and MT-10 where the mean concentrations were

significantly less.

The inorganic analyses of the whole body samples of

Largemouth Bass and Channel Catfish resulted in the detection of

15 and 18 trace and heavy metals, respectively (Table 21 and 22).

Lead, chromium, and arsenic were unique to the Channel Catfish

samples. These elements were limited to only a few observations

among the samples as were cobalt, copper, and nickel for both

species. Those elements common to both the bass and catfish

samples included aluminum, barium, calcium, iron, magnesium,

manganese, potassium, selenium, sodium, vanadium, and mercury.

The range of concentrations for these elements were generally

similar among all stations with the exception of mercury.

The relationship between the whole body concentration of

mercury and fish length was considered through linear regression

analysis (Figures 26 and 27). For both Largemouth Bass and

16
Channel Catfish the small R SQUARE values of 0.077 and 0.038

indicate that mercury concentration was nearly independent of

body length. The distribution of bass and catfish whole body

mercury concentrations among the sampling stations is illustrated

in Figures 28 and 29. With the exception of station MT-2, the

average and range of mercury for the bass remained relatively

consistent among the stations with average concentrations varying

from between 0.11 to 0.26 mg/kg. Mercury concentrations of whole

body bass from station MT-2 averaged about three times greater

than mean values of the other stations. Using Duncan's multiple

range test at an alpha of 0.05, a comparison of the means showed

that average concentrations of mercury were not significantly

different between stations MT-2 and MT-8, a background site but

significantly increased above the means for all other sampling

sites.

Concentrations of mercury for the whole body samples of

catfish appeared more variable among the stations than with the

bass. Mean values for total mercury ranged from 0.04 to 0.74

mg/kq. The latter concentration was associated with station MT-3

whir., was not significantly different from ei_her the background

site at MT-9 or the reference station MT-10, per the Duncan's

multiple range test at an Alpha of 0.05.

The forage fish assemblage sampled in the study was mainly

represented by juvenile menhaden, silversides, small sunfish,

shad and assorted minnows. Results of the inorganic analyses of

the forage fish composites are provided in Table 23. Detectable

concentrations of arsenic, chromium, copper, and lead were found

in less than 20 percent of the samples analyzed (N=29) and were

scattered among the stations. Concentrations of barium,

magnesium, pot^^ium, selenium, sodium, vanadium and mercury were

common to all stations. Mercury concentrations varied from 0.02

to 0.07 mg/kg across all stations. Thirty-three percent of the

forage fish samples analyzed yield MDL results. Of the remaining

elements detected, che- concentrations of aluminum, iron,

manganese, and zinc were generally about 10 times greater in

forage fish than in either Largemouth Bass or Channel Catfish.

The results of the organic analyses of the forage fish

composite samples indicated the presence of one Aroclor compound

which was PCB-1260 and the pesticide DDT, DDE and DDD (Table 24).

The PCB compound was detected in one sample from station MT-0 and

MT-10, a reference site on the Alabama River. The observed

levels of DDE ranged from a MDL of 0.005 to a maximum detectable

concentration of 0.250 mg/kg. DDT concentrations ranged from the

MDL which varied from 0.005 to 0.020 mg/kg to a maximum

detectable level of 0.029 mg/kg. The distribution of total DDT

relative to sampling station is shown in Figure 30. The

calculation of total DDT included median values for all the

observed MDL results. With the exception of station MT-7, the

maximum concentration remained less than 0.10 mg/kg at all

17
stations with the average varying from 0.008 to 0.067 mg/kg. The

concentrations for the three replicate samples of forage fish

from station MT-7 ranged from 0.074 to 0.310 mg/kg and averaged

0.158 mg/kg. This mean, when compared to other stations, was not

significantly different from station MT-2 as determined from

Duncan's multiple range test at an alpha of 0.05. These two

stations both yield mean concentrations significantly greater

than all other stations.

4.2.3 Floodplain Fish

The floodplain wetlands were sampled during the first week

of April, 1994 when the river stage was sufficient to inundated

the area and provide access by boat. Those stations involved in

the sampling included MT-0, MT-2, MT-7, MT-8, and MT-11. Few

specimens of the targeted species were collected. The dispersion

opportunities of the fish in these inundated areas of the

floodplain were greatly enhanced relative to the river when flow

was within bank, thus making the fish more difficult to locate

and capture. Those fish collected included forage fish, from all

stations, Largemouth Bass from all stations ^;>c<-_t MT-7, and

Channel Catfish from only station MT-2. The forage fish were

represented primarily by juvenile shad. All the fish collected

were processed for whole body chemical analyses because of their

few numbers. Also the stomach content of the collected

Largemouth Bass and Channel Catfish were examined and chemically

characterized when sufficient sample biomass was available.

The inorganic analyses of the whole body tissue from the

floodplain wetlands identified 17 trace and heavy metals (Table

25). Of these elements, aluminum, chromium, cobalt, copper,

nickel, and vanadium were found at detectable concentrations in

only the forage samples. Barium and iron were common to the

forage fish but only detected in two bass and one catfish

samples, respectively. The remaining elements of calcium,

magnesium, manganese, potassium, selenium, sodium, zinc, and

mercury were found at detectable concentrations in all fish from

all stations. The range of Largemouth Bass and forage fish whole

body concentrations of this latter group of elements were

comparable to the values reported for the river samples of fish

with the exception of mercury in the forage fish from the

floodplain of stations MT-2 and MT-11. Mercury ranged from 0.02

to 0.07 mg/kg in forage samples from the river when sampled six

months prior to the floodplain sampling (Table 23). The

floodplain composite samples of the forage fish yielded mercury

concentrations of 0.05, 0.05, 0.10, 0.21, and 0.23 mg/kg with the

latter two observations coming from the reference site MT-11 on

the Tensaw River and MT-2 in the Cold Creek Swamp, respectively.

The organic analyses of the whole body fish samples from the

floodplain wetlands indicated the presence of two pesticides at

detectable concentrations. These compounds were rethoxychlor

18
which was found only at station MT-7 and DDT and related

metabolites DDE and DDD (Table 26). The metabolite DDE was

detected at all stations ,in samples of the Largemouth Bass and

forage fish. Concentrations ranged from the'MDL's which varied

from 0.005 to 0.010 mg/kg to a maximum detectable concentration

of 1.20 mg/kg which occurred in the forage fish sample from the

wetland associated with station MT-7 at Mclntosh. The second

greatest concentration was 0.85 mg/kg from stations MT-8, a

background site.

Total DDT roncentrations were at least in order-of-magnitude

greater in the forage fish at MT-7 and 8 than the other

floodplain stations. The 'former location represented the general

floodplain area between the Olin Basin and the Tombigbee River at

Mclntosh. The latter station was a background station located

about 4 river miles upstream from Mclntosh in the Three Rivers

Lake region (Figure 1). Total DDT concentrations of forage fish

samples from the river at these two stations in the November 1993

survey were one to two orders of magnitude less (Table 24 and

26) .

For mercury, concentrations in the forage fish were about

four times greater in the floodplain at stations MT-2 and 7 than

at the other sampling areas (Table 25). As was the case with

DDT, the forage samples collected from the floodplain, contained

10 times the mercury concentrations found in forage samples from

the river. In the case of the few Largemouth Bass collected

maximum concentrations of mercury were observed in samples from

MT-2 and MT-7 which were associated with the Stauffer and Olin

NPL Sites where mercury has been identified as a chemical of

concern. The concentration of mercury in these floodplain

samples of bass were well within the range reported for river

samples at MT-2.

5.0 Fish Health Index Assessment

5.1 Sampling Strategy and Methods

The Fish Health Index (FHI) assessment was initially planned

to involve Largemouth Bass as the predator species of interest.

When the assessment was implemented in the period of October 25

to November 2, 1994, the Channel Catfish was selected as a second

target species. This species was added to the study because of

its popularity as a commercial species and its distribution

throughout the study area.

Largemouth bass were collected by electrofishing. Channel

catfish were collected by electrofishing and cheese baited slat

traps. Fish samples were collected fron the same nine sampling

stations used to collect fillet and who-.e body samples for tissue

analyses (Figure 1). Upon collection in the field, fish were

19
placed in a live well and disturbed as little as possible until

anesthetized for the autopsies. The minimum collectable length

for Largemouth Bass was 250 mm and for Channel Catfish 325 mm.

An attempt was made to collect 15 bass and 15 catfish at each

sampling location. Internal and external examinations were

performed on over 200 individual fish according to EPA Region IV

protocol for conducting a Fish Health Index (FHI) (Appendix D).

All autopsies were performed in the field in a mobile

laboratory. In general the autopsy consisted of an external and

internal examination and a blood analyses. Observations were

recorded on field data sheets as a number or letter code. The

codes used on field data sheets were the same codes used by Goede

(1993). Copies of field data sheets are included in this report

in Appendix F.

After completing autopsies, the number/letter code

describing the physical/pathological condition of each tissue or

organ examined was assigned a numerical score (except for sex,

fat deposits, and the gall bladder). Next, an accumulative score

for each fish was calculated and then a mean FHI score was

calculated for groups of fish from the same sampling station.

All scoring of physical/pathological conditions and subsequent

calculations were performed automatically upon entering field

data into a computerized spreadsheet. The spreadsheet was

designed specifically to manipulate fish health data. Some of

the number/letter codes on the field data sheets had to be

replaced with a new code number/letter before entering on the

spreadsheet before manipulations and calculations could be

executed. The new codes are listed in the protocol for conducting

the FHI. Spreadsheets reflecting these code changes as well as

displaying the calculated scores mentioned above are included in

this document as Appendix E.

5.2 Autopsy Results

A total of 218 fish were collected from 9 sampling stations

on the Mobile, Tombigbee, and Alabama Rivers (121 largemouth bass

and 97 channel catfish). At least 10 fish of each species were

collected at each station except for catfish. Only four catfish

were collected at station MT-4 and no catfish were collected at

station MT-10. Autopsies were performed on all 218 fish and the

condition of their tissues and organs are summarized in the field

data sheets and spreadsheets included in Appendix B and Appendix

C respectively. Note that calculated scores for catfish did not

include blood data because criteria for determining "normal" and

"abnormal" conditions of blood in this fish species had not been

established at the time this study was conducted. The FHI scores

for fish collected at each sampling station are summarized in

Tables 27 and 28.

Externally all fish appeared healthy. All fish had good

20
color and, except for an occasional lesion or external parasite

("Anchor Worm"), all fish had healthy-looking skin, fins, and

opercula.

Internally, fish from all stations showed, to some degree,

abnormalities of the liver, spleen, and/or kidney. The primary

abnormality encountered was the presence of nodules. In

Largemouth Bass all three organs were usually affected. In

catfish the nodules were restricted to the liver. Dissection of

several liver nodules produced encysted parasitic worms in both

bass and catfish. Discoloration of the liver was the second most

frequently encountered abnormality, especially in bass. No gross

abnormalities were detected in the blood of either fish species.

In general, the health of the Largemouth Bass was poorer at

the upper and lower reaches of the study area, including the

background (MT-9) and reference (MT-10) stations (Figure 31).

The healthiest bass (i.e. lowest score) were found at station MT5. The mean FHI score for this station was significantly better

(statistically) than the mean score for the reference station.

Overall, the FHI scores for catfish were lower (i.e. better

health) than those for bass. The highest score (i.e. poorest

health) was generated by catfish from station MT-3. However,

this score is not significantly different (statistically) from

the score for the background station MT-9.

6.0 Discussion and Review of Results

The analytical approach to this' study involved the analyses

of water, sediment, and fish tissue for numerous organic

compounds and metals. Of this number, 39 were identified as

chemicals of concern because of their known association with the

four NPL Sites. The results of the chemical analyses identified

the presence of only a few of these selected chemicals and

elements. Of the more limited array, mercury and the pesticide

DDT and related metabolites DDE and DDD emerged as the most

salient constituents of environmental concern.

Through the screening study efforts, these chemical

constituents were found in elevated concentrations, relative to

background and reference conditions, in sediments and surface

water in close proximity to the NPL Sites. These areas were the

floodplain wetlands of Cold Creek Swamp where concentrations of

mercury in water and sediments were increased many times above

background and reference conditions. To a lesser extent, the

same scenario for mercury applied to the floodplain wetlands

associated with the Olin Basin. The pesticide DDT and related

metabolites were also detected in the fioodplain and river

sediments closely associated with the Olin Basin but not

elsewhere in the study's sampling regime. Based upon these

21
findings, the NPL Sites were recognized as source factors of

contamination to be considered in the assessment of public health

and ecology associated with the river.

Of the pesticide DDT and related metabolites, DDF was found

in measurable concentrations in nearly all of the fillet samples

of Largemouth Bass and Channel Catfish. The compound DDD was

detected in fillet samples. DDT was limited to fillet samples

from station MT-7 at Mclntosh. Total DDT concentrations of the

composited fillet samples were substantially less than the action

level of 5 mg/kg of the USFDA.

The fish tissue criteria promulgated by EPA for the

protection of human health is considerably more stringent than

either the USFDA action level or the Alabama criteria. EPA's

criteria is specific to DDT, DDE, and DDD and values of 0.0316,

0.0316, and 0.0449 mg/kg, respectively. Relative to DDT and DDD,

the observed concentrations exceeding criteria were limited to

fillet samples from the area of station MT-7 at Mclntosh.

Concentrations of DDE exceeding criteria were detected in samples

from all stations except MT-10 which was a re: -~ rence station on

the Alabama River near Chrysler, Alabama. The cumulative

frequency distribution (CDF) of fillet samples of DDE exceeding

the EPA criteria of 0.0316 mg/kg in Largemouth Bass and Channel

Catfish numbered approximately 25 and 50 percent of the samples,

respectively (Figures 32 and 33). Of those samples exceeding the

EPA criteria, their distribution remained central to station MT-7

at Mclntosh where the Olin and Ciba-Geigy NPL Sites are located

(Figures 11, 12, and 13). This central tendency m sample

distribution was also validated by demonstrating that the mean

concentration of total DDT in the fillet samples of catfish were

significantly greater at station MT-7 than all other stations

(Table 29).

The greatest observed mean whole body concentration of total

DDT for Largemouth Bass was from station MT-7. The

concentration, however, was not significantly different from the

downstream concentrations nor the background stations MT-8 and

MT-9. All whole body mean concentrations for bass from the

Mobile/Tombigbee River were significantly greater than samples

from the reference station MT-10 of the Alabama River. Similarly

for Channel Catfish, the mean whole body concentration of total

DDT for station MT-7 was the largest but not significantly

different from the down stream stations and reference station MT9. The lowest mean concentrations of total DDT in whole body

catfish were associated with the background station MT-8 and the

reference station MT-10. The mean concentration of total DDT

found in the forage fish samples was significantly greater at

station MT-7 and MT-2 when compared to all other river stations.

Total DDT concentrations were found at even greater levels in the

forage community sampled in the inundated flcodplain wetlands

associated with station MT-7 and the upstr^-an background sampling
area of station MT-8.

Based upon the above findings involving fillets and whole

body levels of DDT and related metabolites DDE and DDD, the

sampling area of station MT-7 at Mclntosh appears to be a factor

consistently associated with elevated concentrations of these

compounds as indicated in Figures 11, 12, 13, 24, 25 and 30. To

a lesser extent, the two background stations (MT-8 and 9) were

also associated with elevated levels of these pesticides. The

reference sites MT-10 on'the Alabama River and MT-11 in the

floodplain in the Tensaw Lake area yielded samples that

consistently featured the smallest concentrations of these

compounds. Since the two background stations (MT-8 and MT-9)

were located about four to five river miles upstream of the NPL

sites at Mclntosh, the mobility of fish could be a consideration

in explaining the pattern of contamination in the upper section

of the study reach. Results of mark and capture studies

involving Largemouth Bass in the Mobile River system may not

strongly support this consideration. Bass captured from the

delta area of the Mobile River were tagged and released and

rec..pt u.re. The recaptured bass showed relat1'. ^ly little movement

from their original location of capture and release

(Tucker,1994). No evidence of sediment or surface water

contamination was apparent at the upstream reference stations.

The only evidence of DDT and metabolite contamination occurred in

sediment samples from the Mclntosh floodplain wetlands and in the

river where the sampling areas interface with the Olin Basin.

The movement of some fish between the river and basin is

probable because of surface water connections between .the two

water bodies. Studies reported by Woodward and Clyde, 1994,

indicate Largemouth Bass, Mosquito Fish, and oth?r aquatic

species in the Basin are excessively contaminated with mercury

and DDT and its associated metabolites. The immigration of

contaminated fish such as Largemouth Bass from the basin to the

river and their mingling with the river population remains a

plausible explanation of the fish tissue contamination. The fish

fillet samples analyzed in this study were derived from a

composite of five fillets. Each fillet was obtained from

different fish of a similar size. Hence, a single fillet from

one highly contaminated fish which may. have originated from the

Basin could render the composite as excessively contaminated.

The composite samples represents an average concentration. An

alternate and less apparent explanation of the contaminated

composite fillet samples in the Mclntosh area could rest with

unknown sources upstream of station MT-7.

If the NPL sites at Mclntosh are viewed as a source of DDT,

DDE, and DDD to the river, their effects appear to be diminishing

in view of the findings of a study conducted in 1990 by the U.S.

Fish and Wildlife Service. In this investigation, sampling was

conducted to collect samples of the Largemouth Bass population in

23
the area of the Tombigbee River associated with Mclntosh and the

Olin and Ciba-Geigy NPL Sites. In their sampling, 15 bass were

collected and processed for fillet and whole body analyses.

These analyses included the determination of concentrations for

DDT, DDE, DDD, and mercury.

Total DDT was reported as the sum of the three compounds

which in the whole body samples ranged from 0.34 to 34.53 mg/kg

and averaged 8.0 mg/kg. Fillet concentrations of total DDT

ranged from 0.08 to 2.22 mg/kg and averaged 0.70 mg/kg. The mean

concentrations reported in the current study for total DDT in

whole body and riliet samples of Largemouth Bass from MT-7 were

0.50 and 0.30 mg/kg, respectively (Table 29). A comparison of

the mean values for the two studies will suggest a significant

decrease in the contamination level of the Largemouth Bass has

occurred over the last three years. These data when coupled with

.the NCBP historical data referenced in the same 1990 USFWS report

suggest even a more long term decrease in pesticide

concentrations (Figure 34). The referenced 1990 USFWS report also

provided some historical mercury concentrations for whole body

and f^let samples of Largemouth Bass from the- subject area. A

comparison of these data with bass results from MT-7 of the

current study would also suggest a decline in mercury

contamination (Figure 35).

Relative to public health considerations, concentrations of

mercury in the fillet samples for both the Largemouth Bass and

Channel Catfish were all less than the action level of 1 mg/kg as

supported by the USFDA and EPA, respectively (Table 30). Mercury

concentrations of the fillet samples of the bass correlated

fairly well with fish total length; hence, the fillets of larger

fish contained greater concentrations of the metal. Mercury

concentrations for the fillet samples of large bass were not to

comparable among sampling stations because of too much variation

about the means. In contrast, the range of concentrations in the

fillet samples of catfish were relatively narrow and very

comparable among the sampling stations except for MT-2 where the

river interfaced with the lower floodplain of Cold Creek Swamp.

The averaged concentration or mercury in the samples of catfish

fillets from MT-2 was twice the values of the other stations and

shown to be significantly different.

The mean whole body concentrations of mercury in Largemouth

Bass between sampling stations were not significantly different

between stations MT-8 and station MT-2. Samples from these two

stations yielded mean concentrations significantly increased

above all other stations. For Channel Catfish, the mean

differences between the stations were not significant.

Concentrations of mercury among the forage fish samples of the

river were low compared to background and reference stations.

Based upon the findings for mercury, station MT-2 emerged as

24
the only sampling location in the study area where concentrations

of this metal were found at levels significantly elevated over

all other stations including both background and reference sites.

This occurred in fillet samples of Channel Catfish. The mean

concentration, of mercury of whole body samples of Largemouth Bass

from station MT-2 were also significantly increased above all

other station except the background site MT-8.

Station MT-2 was established in an area of the Mobile River

adjoining the floodplain wetlands of the Cold Creek Swamp and the

associated Stauffer NPL site. A surface water connection occurs

between the river and swamp during some periods of high river

flow, thus providing a hydrologic pathway for exchange of biota

and mercury contaminated surface water. Surface water

concentrations of mercury in the floodplain wetland when

inundated were found to exceed EPA.surface water criteria.

Mercury concentration reported for both the catfish and bass

reflect the probable effects of this connection.

The Fish Health Index (FHI) results indicate that both the

La_-gemouth Bass and Channel Catfish were in relatively good

condition with the exception of encysted parasitic worms in the

spleens, livers and kidneys which was common to all samples.

Base upon the statistics of the FHI protocol, bass from station

MT-5 were in the best condition. The average concentration of

mercury in either the fillets -of whole body samples of bass

ranked least among the sampling stations. Station MT-5 was

located in the Tombigbee section of the study area just upstream

of the confluence of the Alabama River. In terms of the-FHI, the

Channel Catfish were found in better condition relative to the

bass. Also, the mean concentrations of mercury in whole body

samples of catfish were least when compared to the other

stations.

The consequences of the reported fish tissue contamination

can be considered in terms of public health and ecological

effects. Regarding the former consideration, mercury did not

pose a concern because the observed levels were less than the

action level of the USFDA or the criteria of EPA. The same

conclusion could also be reached in terms of total DDT relative

to the USFDA action level of 5 mg/kg or possibly the Alabama

criteria which was exceeded 10 and 7 percent of the Largemouth

Bass and Channel Catfish fillet samples. The concern for public

health is heighten somewhat on the basis of DDT, DDE, and DDD

concentrations exceeding EPA criteria in 26 and 53 percent of the

fillet samples of Largemouth Bass and Channel Catfish,

respectively.

6.1 Public Health Considerations

Based upon the reported con' entrations of mercury and DDT

and associated metabolites DDE e .a DDD, a human health risk
assessment was conducted (Appendix G). A summary of the risk

assessment findings are provided in Table 32. In this

assessment, carcinogenic and non-carcinogenic risks are

considered. The carcinogenic evaluation relates to the

probability of an individual incurring incremental cancer risk

due to the consumption of fish at a given frequency, quantity,

and concentration of DDT contamination. For toxic or noncarcinogenic effects, a hazard quotient (HQ) is also calculated

on the basis of consumption of fish with a given frequency,

quantity, and concentration of DDT or mercury, the assumptions

regarding these variables are given in the risk assessment of

Appendix G. A HQ greater than one indicates that exposure to the

contaminant will likely cause adverse effects. Tissue

concentrations used in this risk assessment were based upon

analyses of fish fillet samples from stations MT-2 and MT-7

because these stations yielded samples with the highest average

'and maximum levels of mercury and total DDT, respectively. The

maximum reported concentrations actually constitute and average

of five individual fillets that were composited and subsampled

for analyses.

Results of this assessment indicate that a person consuming

bass or catfish fillets from station MT-7 risks an additional low

to moderate chance (one chance in 10,000 to 100,000) to incur a

cancer. Non-carcinogenic effects from consuming catfish from

non-carcinogenic are unlikely. The HQ for Largemouth Bass is

higher and indicates an increased potential for adverse effects.

The consumption of Largemouth Bass or Channel Catfish

fillets from station MT-2 would result in exposure to

concentrations of mercury that are at or below levels shown to

cause adverse toxic effects.

6.2 Preliminary Ecological Risk Evaluation

In order to determine whether data collected in this study

may warrant pursuit of a full ecological risk assessment, a few

exposure scenarios were compiled into a preliminary ecological

risk evaluation. This evaluation draws on relevant information

presented recently for other Region IV sites in an ecological

risk assessment (US EPA 1994a) and another preliminary ecological

risk evaluation (US EPA 1994b).

6.2.1 Assumptions

This evaluation concentrates on the dietary exposure of area

wildlife to the two primary contaminants of concern for the

riverine segment studied, total mercury and DDTr. The exposure

scenarios chosen maximize fc^dchain exposure through ingestion of

fish; the focus of this study. The wood stork (Mycteria

americana) and the belted kingfisher Ceryle alcyon) will serve

26
as receptors for this evaluation. Both are upper trophic level

receptors for food chain exposure pathways that may exist at this

site. The diet of each species consists primarily of fish (US

EPA 1993, US EPA 1994b). The wood stork is a Federally listed

(June 3, 1994 amendment) endangered species statewide in Alabama.

This species also serves as a representative receptor of other

large fish eating wading birds sharing a similar feeding regimen.

The belted kingfisher has as a permanent range Alabama and much

of the southeast (Audubon 1994) and is an often seen resident

within the study area.

An area use factor of 1 will be used in this evaluation.

This value was chosen since the feeding range of both species

could be easily contained within the 53 mile segment of the

rivers studied. The range of the stork is approximately 20 miles

(32 km, US EPA 1994b), and the shoreline range of the kingfisher

has a mean of up to 1.4 miles (2.2 km, Brooks and Davis 1987 as

cited in US EPA 1993). With these ranges, the birds could feed

exclusively along the river and not extend beyond the segment

studied.

Estimated dose calculations.assume foraging beyond the

bounds of any one sample station, and use, therefore, a mean

concentration of total mercury or DDTr for all river stations

(MT-0 to MT-9) other than the reference station MT-10. In

determining this mean concentration, all stations for -which there

were either a confirmed, estimated (analytical code "J"), or

presumed, (analytical code "N"), values were used in averaging.

Samples below the limit of detection (analytical code "U") were

used in' the averaging calculations by assuming a median

concentration for the reported MDL. When determining the mean

for DDTr, station means were first determined, then a mear

concentration was determined for all river study stations. For

the purposes of this evaluation, it was assumed that the

toxicities of DDT, DDE and DDD were additive. It was also

assumed that mercury and DDTr were 100 percent available to the

organism from their prey.

Chronically toxic levels of mercury and DDTr were not found

for the species used in this evaluation, however, a No Observable

Adverse Effect Level (NOAEL) for each contaminant for piscivorous

birds that has been used in other recent evaluations (US EPAa,b)

was used here.

6.2.2 Exposure Pathways

The exposure pathway selected to determine risk to the stork

and the kingfisher is through the ingestion of contaminated

forage fish. It was assumed that forage fish, defined in this

study as any fish captured during electrofishing that was no

longer than three inches, total length, comprised 100 percent of

the diet of these piscivorous birds. An assumption of this

27
approach is that exposure calculated from collected biota is

representative of actual exposure. Exposure from incidental

water or sediment ingestion was not included here. It is

expected that such additional exposure would be minimal for the

kingfisher. For the stork, sediments entrained during shallow

water foraging may constitute a more important source of dietary

contaminants.

6.2.3 Exposure Profiles

6.2.3.1 Wood Stork (US EPA 1994b)

The wood stork is the only native stork in North America

north of Mexico. It is more or less resident of Florida and

Georgia, very rarely elsewhere along the coast from South

Carolina to Texas (Audubon 1994). It is currently Federally

.listed as an endangered species for the entire state of Alabama

(June 3, 1994 listing amendment). This is primarily a freshwater

bird. It is, however, frequently observed in saltwater marshes

(Eckert 1976) . Although sightings are rare in the study area

this ;p.ecies is used hrre because of its endan cred status in <-lie

state, its heavy dietary reliance on fish and it's upper level

trophic position, all of which make it a reasonable candidate for

a worst case risk scenario. Though primarily a freshwater bird,

it is frequently observed in saltwater marshes.

This is a gregarious species and is rarely seen alone.

Storks are usually seen in groups or with other wading birds,

eating, roosting, nesting, migrating, and performing all other

activities (Eckert 1976) .

Adult storks weigh 2050 grams (Dunning 1993), and consume

520 grains of food per day (Kahl 1964) . Dietary composition is

mostly fish, however, wood storks eat a wide variety of small

prey including frogs, water snakes, lizards, minnows, wood rats,

fiddler crabs, turtles, tadpoles, and water beetles (Eckert

1976). Since fish comprise most of the stork diet, and since

this study has provided current information on forage fish

contaminant content, calculations assume forage fish constitute

100 percent of the stork diet.

6.2.3.2 Belted Kingfisher (US EPA 1994a)

The belted kingfisher is a medium-sized bird that eats

primarily fish. It is one of the few species of fish-eating

birds found throughout inland areas as well as coastal areas.

The belted kingfisher's range includes most of the North American

continent; it breeds from northern Alaska and central Labrador

southward to the southern border of the United States (US EPA

1993). Alabama and essentially the entire southeast are part of

its permanent range (Audubon 1994) .

28
Adult kingfishers weigh from 113 to 178 grams (Fry and Fry

1992). Reported ingestion rates range from 0.5 (Alexander 1977)

to 1.75 (White 1936) times the bodyweight per day. For this

evaluation, a body weight of 113 grams and ingestion rate of

0.198 kg per day (1.75 cimes a bodyweight of 113 grams) will be

used. The diet is predominantly fish but includes frogs crayfish

and aquatic reptiles (Fry 1980). For this evaluation, it was

assumed that the diet was 100 percent fish. Feeding range while

nesting is 8050 acres (Cornwell 1963).

6.2.4 Effects Profiles

6.2.4.1 Mercury (US EPA 1994b)

Several studies were located that evaluated dietary mercury

exposure to predatory birds, both raptors and piscivorous

species. Several laboratory feeding studies conducted on

goshawks and red-tailed hawks indicate that these species

accumulated mercury and exhibited toxicity symptoms when fed prey

containing elevated mercury levels. A field rtudv conducted on

the -_orr^non loon also, indicated that this specj.es -xhibits

toxicity symptoms after ingesting prey with elevated mercury

levels.

Goshawks were fed a diet of chickens which had been fed

methyl mercury-dressed wheat (0.4 - 0.5 mg/kg/day) for five to

six weeks and both chickens and goshawks were sacrificed (Borg et

al. , 1970) . All chickens were clinically healthy at the end of

the feeding period. The average mercury level in the chicken

feed was 8 mg/kg, resulting in skeletal muscle concentrations of

10 mg/kg. Muscle and liver from the chickens was fed to

goshawks. Intake of Hg by the goshawks was 0.7 - 1.2 mg/kg/day.

Clinical symptoms of Hg poisoning appeared after two weeks. All

birds were dead 47 days after the start of the experiment.

Muscle Hg levels of the goshawks averaged 40 - 50 mg/kg,

representing a concentration factor of 4 - 5 in the second link

of the food chain. Brain mercury levels in the dead goshawks

ranged from 30 to 40 mg/kg.

Red-tailed hawks were fed chickens contaminated with methyl

mercury (Fimreite and Karstad, 1971). The chicks were fed diets

containing Panogen 15, a commercial seed treatment containing 2.5

percent methyl mercury dicyandiamide (MMD) at rates of 6, 12, and

18 mg/kg MMD for three weeks. Mercury levels measured in chick

livers were 3.9, 7.2 and 10.0 mg/kg, respectively. Mean

estimated intake of mercury by the three groups of hawks over the

12-week exposure period was 0.575, 1.12 and 1.46 mg mercury per

day, respectively. Mortality occurred in hawks receiving the

most contaminated diet (1.12 mg/kg/day) after an exposure period

of one month or more. Pathological changes noted in all hawks

which received the highest mercury doses included swelling of

axons of myelinated nerves in the t.punel cord, and dilatation of

29
myelin sheaths and loss of myelin.

Barr (1986) noted decreased reproductive success and

behavioral changes in common loons nesting on the WabigoonEnglish River systems, areas affected by unpredictable water

level fluctuations and Hg contamination. Water level

fluctuations due to dams were ruled out as the cause, as the

decreased nesting success observed both in lakes experiencing

only natural water level changes, as well as in areas affected by

dams. A strong negative correlation was found between the

successful use of territories by breeding loons and mercury

contamination. A reduction in egg laying, and nest site and

territorial fidelity, were associated with mean mercury

concentrations ranging from 0.3 to 0.4 mg/kg in prey (equivalent

to 0.1 mg/kg body weight/day), and from 2 to 3 mg/kg in adult

brain tissue and eggs. Loons established few territories, laid

• only one egg, and raised no young where mean mercury in prey

species exceeded 0.4 mg/kg. Non-mercury toxicants were found in

loons and prey items at low levels, and were therefore discounted

as a major factor in the failure of loon reproduction.

Wood storks feed through the use of tactile response and

exceptionally fast reflex action of the stork. Therefore, subtle

behavioral and/or neurological effects could have substantial

impacts on individual wood storks feeding at this site.

For purposes of this evaluation, dietary levels of 0.03 mg

mercury per kg in prey (0.01 mg/kg/day) will be used as a NOAEL

for the wood stork.

6.2.4.2 DDTr (US EPA 1994a)

DDTr is of particular concern in ecological systems because

of its tendency to accumulate in biological tissues, posing food

chain threats to higher order consumers.

To evaluate the effects of DDTr in piscivorous birds,

Anderson et al. (1975) studied the reproductive success of brown

pelicans off the southern California coast from 1969 to 1975.

Concentrations of DDTr in anchovies (the major food source of the

pelican colony) and pelican eggs were measured. Over the five

year monitoring period, concentrations of DDTr in anchovies

steadily declined from 4.27 mg/kg wet weight in 1969 to 0.15

mg/kg in 1974. Eggshell thickness gradually increased over the

5-year period. Productivity at the pelican colony increased from

a total of four young fledged in 1969 to 1185 fledged in 1974.

At tissue levels of 0.15 mg/kg DDTr in anchovies, the fledgling

rate was 30 percent below the estimated rate necessary to

maintain a stationary population.

Twelve pairs of American kestrels were fed a diet containing

2.8 mg/kg p,p'-DDE on a wet weight basis (Porter and Weimeyer

30
1972). Two of the males died after 14 and 16 months on the diet.

DDE residues in brain tissue were high, and fat reserves were

depleted. The kestrels died at a time when the birds lose weight

and deplete fat reserves due to reproduction and molt.

With regard to the effects of DDTr on eggs and reproduction,

a number of studies correlated dietary levels with levels

measured in eggs. Many field studies (Greichus et al. 1973, Fox

et al. 1980, and Cooke et al. 1976) have reported DDTr levels

measured in eggs and observed adverse effects on reproduction

such as broken eggs. However, few studies report levels of

contaminants in food organisms which contribute to the levels

observed in eggs. Only two studies were located that reported

dietary DDTr levels and levels measured in eggs. One study was

conducted with a raptor, and the second with chickens.

American kestrels were fed diets containing 2.8 mg/kg p,p'-

DDE on a wet weight basis for 2 years (Weimeyer and Porter .1970) .

The DDE was dissolved in cottonseed oil and added to ground meat.

The first egg laid by each pair was collected in 1968 and again

in 1969. Levels of PIP'-DDE measured in eggs ranged from 1.05 to

5.89 mg/kg in 1968, and from 17.4 to 44.2 mg/kg in 1969. Average

eggshell thickness was 10 percent less in 1969 than in 1968.

Smith et al., (1970) fed laying hens diets containing DDT at

levels of 0, 0.05,. 0.5 and 5.0 mg/kg for 2 months. DDT levels in

eggs were measured before and after force molting. Levels in

eggs before the molt were not elevated prior to the molt,

although levels in abdominal fat were. After the forced molt,

levels in eggs, for hens consuming the dietary levels reported

above, were 0.1, 1.2, 4.1 and 11.9 mg/kg, respectively.

Also with regard to the effects of DDTr on eggs and

reproduction, a number of studies have correlated percent shell

thinning with egg loss. Several of these have correlated

measured DDT residues in eggs of birds to increased thinning and

egg loss due to breakage. Eggshell thinning of greater than 20

percent appears to result in decreased nesting success due to

eggshell breakage (Anderson and Hickey 1972, Dilworth et al.

1972). Predatory and piscivorous birds appear to be at greatest

risk for accumulating levels of DDTr which will result in this

degree of eggshell thinning.

Lincer (1975) conducted laboratory feeding studies with

American kestrels in which females were maintained on diets

containing 0.3, 3.0, 6.0 and 10.0 mg/kg DDE (0.054, 0.55, 1.1 and

1.83 mg/kg body weight (BW) per day, respectively). Animals on

the 0.3 mg/kg diet produced eggs with shells which were not

significantly different in thickness from controls, and 6 percent

thinner than archived eggshells collected prior to widespread use

of DDT. Birds maintained on the 3.0 and 6.0 mg/kg diets produced

eggshells which were 14.0 and 17.4 percent thinner than

31
experimental controls, and 25.5 and 28.3 percent thinner than

pre-DDT eggshells, respectively. Eggshell thinning to this

extent may be expected to affect breeding success. While the

kestrel is not a piscivorous bird, it feeds at a similar trophic

level.

A NOAEL of 0.054 mg/kgBW/day were used to evaluate the

effects of DDTr on piscivorous birds for this evaluation.

6.2.5 Risk Characterization

To estimate risk to the wood stork and belted kingfisher

along the Mobile River study site, implications of the exposure

concentrations need to be determined. The hazard quotient method

(US EPA 1989, Barnthouse et al., 1986) compares exposure

concentrations to ecological endpoints such as reproductive

failure or reduced growth. The comparisons are expressed as

ratios of potential intake values to population effect levels,

or:

Hazard Quotient = Exposure Concentration

No Observed Adverse Effect Level (NOAEL)

A hazard quotient greater than one indicates that exposure to the

contaminant will likely cause adverse effects in the organism. A

hazard quotient less than one does not necessarily indicate a

lack of risk, but should be interpreted based on the nature of

the effect used as the denominator, and the magnitude of the

calculated quotient.

6.2.6 Sources of Uncertainty

There are factors inherent in the risk assessment process

which contribute to uncertainty and need to be considered when

interpreting results. Major sources of uncertainty include

natural variability, error, and insufficient knowledge.

Risk (hazard quotient) calculations were based on

conservative values; the lowest body weight and highest ingestion

rate cited for the stork and kingfisher were used in the exposure

profile and the NOAELS used to determine hazard quotients were

the lowest values found in the literature

An important contributor to uncertainty is the

incompleteness of the data or information upon which the risk

assessment is based. Literature values of life history

information used in the exposure profile may not be truly

representative of values for the stork and kingfisher in southern

Alabama. Although the reported NOAELs used in the quotient

calculations are generally for closely related species, response

to mercury and DDT for the wood stork and belted kingfisher may

be different than in species for which data was available. Doses

32
reported in toxicological studies were in units of mg

contaminant/kg diet, or in units of mg contaminant/kg body

weight/day. Doses were converted to units of mg/kg/day using

reported average body weights and ingestion rates.

NOAELs obtained from the literature may over- or underestimate actual values for the species addressed in this

evaluation. Another source of uncertainty arises because

toxicity values reported in the literature are often derived in

single species, single contaminant laboratory studies.

Prediction of ecosystem effects from laboratory studies is

difficult, as environmental factors and interactions between

contaminants in field conditions can influence a toxins effects.

6.2.7 Results and Conclusions

This screening level evaluation indicates that there is

reason to believe that piscivorous birds that rely on this 53

mile stretch of the Mobile River as a primary food source may be

at risk from feeding on forage-size fish. In t-.he four risk

seen?.ics developed for the wood stork and the kingfisher exposed

to mercury and DDTr individually and from food sources only,

hazard quotients all ranged well within an order of magnitude of

1.0. For the wood stork, a HQ of 0.06 was calculated for

exposure to mercury (Table 33), and a HQ of 0.277 was calculated

for exposure to DDTr (Table 34). For the belted kingfisher,

whose feeding rate per unit body weight is approximately seven

times higher than that of the wood stork, therefore increasing

exposure through this route proportionally, a HQ of 3.85 was

calculated for exposure to mercury (Table 35), and a HQ of 1.91

was calculated for exposure to DDTr (Table 36).

With all four calculated values within an order of magnitude

of 1.0, it would be advisable to develop a comprehensive

ecological risk assessment for this study area. This would

provide an opportunicy to fully explore exposure of animals at,

relative to, or suspected of inhabiting this area through a full

spectrum of scenarios to determine whether a significant hazard

is posed by this stretch of the river to that biota. Changes in

assumptions would be expected to move HQs in either direction.

Fish or mammals, having different exposure scenarios and toxic

responses to mercury and DDT will have different HQ values for

this geographical area. If an area use factor other than one

were used, for instance, to develop a worst-case feeding

scenario by centering the feeding range on the sample locations

that produced fish with the highest body concentrations of

mercury and DDT, the HQs calculated could increase considerably.

This would be especially true in the case of the kingfisher,

having a much more restricted range than the wood stork. For

instance, if the kingfisher's range centered on the Mclntosh

sample station where mean DDTr in forage fish was 0.158 mg/kg

rather than the overall river reach average 0.059 mg/kg, the
bird's hazard quotient would increase from 1.91 to 5.16, other

assumptions remaining unchanged.

Exposure scenarios for raptors capable of feeding on larger

fish should also be considered. A full risk assessment would

consider exposure from other routes as well, probably increasing

HQ values. Forage ranges that are considered radially from a

nest site rather than linearly along the river, may suggest area

use factors less than one, which would have an effect on the

calculated Hqs. If ranges were extended away from the river, use

should be made of the existing contaminant data that has been

generated for various media from specific industrial floodplain

and on-site locations rather than setting all non-riverine

exposure to zero.

34
7.0 References

Alabama. 1994. Revised Regulation 335-6-10-07. Alabama

Department of Environmental Management. Montgomery,

Alabama.

Alexander, G. 1977. "Food of vertebrate predators of trout

waters in north central lower Michigan." Michigan

Academician. 10:181-195 (as cited in Newell 1987).

Anderson, D.W. -.id J.J. Hickey. 1972. "Eggshell changes in

certain North American birds." In: K.H. Voous, ed.

Proceedings: XV International Ornithological Congress. The

Hague. Netherlands. pp. 514-540.

Anderson, D.W., J.R. Jehl, R.W. Risebrough, L.A. Woods, L.R.

Deweese and W.G. Edgecomb. 1975. "Brown pelicans:

Improved reproduction off the southern California coast."

Science. 190:806-808.

Barnthc..J, L.W., G.W. Sutor, II, S.M. Bartell, J.J. Beauchamp,

R.H. Gardner, E. Linder, R.V. O'Neill and A.E. Rosen. 1986.

User's Manual for Ecological Risk Assessment. Environmental

Sciences Division, Oak Ridge National Laboratory, Oak Ridge,

TN. Publication No. 2679, ORNL-6251.

Barr, J.F. 1986. Population Dynamics of the Common Loon (Gavia

immer) Associated with Mercury-Contaminated Maters in

Northwestern Ontario. Canadian Wildlife Service, Occasional

Paper Number 56.

Borg, K., K. Erne, E. Hanko, and H. Wanntorp. 1970.

Experimental Secondary Methylmercury Poisoning in the

Goshawk (Accipiter g. gentilis L) . Environ. Pollut. 1:91-

104.

Brooks, R.P.; Davis, W.J. 1987. Habitat selection by breeding

belted kingfishers (Cerv'e alcyon). Am. Midi. Nat. 117:63-

70.

Cooke, A.S., A.A. Bell and I. Prestt. 1976. "Eggshell

characteristics and incidence'of shell breakage for Grey

Herons Ardea cinerea exposed to environmental pollutants."

Environ. Pollut. 11:59-84.

Cornwell, G.W. 1963. "Observations on the breeding biology and

behavior of a nesting population of belted kingfishers."

Condor. 65:426-431.

Dilworth, T.G., J.A. Keith, P.A. Pearce and L.M. Reynolds. 1972.

"DDE and eggshell thickness in Nev. Brunswick woodcock." J.

ofWiidl. Manage. 36 (4) : 1186-]i: '
Dunning, John B. 1993. CRC Handbook of Avian Body Masses.

Press, Boca Raton, FL.

CRC

Eckert, Allan W. 1976.

(North of Mexico) .

The Wading Birds of North American

Weathervane Book, New York.

Fimreite, M. and L. Karstad. 1971. Effects of Dietary Methyl

Mercury on Red-Tailed Hawks. J. Wildl. Manage. 35(2):293-

300.

Fox, G.A., K.S. Yonge and S.G. Sealy. 1980. "Breeding

performance, pollutant burden and eggshell thinning in

Common Loons Gavia immer nesting on a Boreal forest lake."

Ornis Scandinavica. 11:243-248.

Fry, C. 1980. "The evolutionary biology of kingfishers

(Alcedinidea)." In: The Living Bird, 1979-1980. The

Laboratory of Ornithology, Cornell University, Ithaca. pp.

113-160.

Fry, C.H. and K. Fry. 1992. Kingfishers, Bee-eaters and

Rollers. A Handbook. Princeton University Press,

Princeton, New Jersey.

Greichus, Y.A., A. Greichus, and R.J. Emerick. 1973.

"Insecticides, polychlorinated bypenyls, and mercury in wild

cormorants, pelicans, their eggs, food and environment."

Bull. Environm. Contain. Toxicol. 9 (6)-.321-328 .

Grover, R. 1989. Environmental Chemistry of Herbicides. Vol.1.

CRC Press.

Kahl, M. Philip. 1964. Food Ecology of the Wood Stork (Mycteria

americana) in Florida. Ecological Monographs 34 (2):97-117.

Lincer, J.L. 1975. "DDE-induced eggshell-thinning in the

American kestrel: a comparison of the field situation and

laboratory results." J. Appl. Ecol. 12:781-793.

Long, Edward R. and Lee G. Morgan, 1990. The potential for

biological effects of sediment-sorbed contaminants tested in

the national status and trends program. NOAA Technical

Memorandum NOSOMA52. National Oceanic and Atmospheric

Administration. Seattle, Washington.

National Audubon Society. 1994. Field Guide to North American

Birds Eastern Region. Ed. John Bell, John Farrand, Jr.

Published by Alfred A. Knopf. New York.

36
Noakes, Scott. 1993. Mobile River Sediment Study. Center for

Applied Isotope Studies. University of Georgia, Athens,

Georgia.

Porter, R.D. and S.N. Wiemeyer. 1972. "DDE at low dietary

levels kills captive American kestrels." Bull. Environ.

Contam. Toxicol. 8(4j :193-199.

Smith, S.I., C.W. Weber and B.L. Reid. 1970. "Dietary

pesticides and contamination of yolks and abdominal fat of

laying hens." Poultry Science. 49:233-237.

Schmitt, C.J., J.L. Zajicek, and P.H. Peterman. 1990. National

Contamination Biomonitoring Program: Residues of

Organochlorine Chemicals in U.S. Freshwater Fish, 1976-1984.

Archives of Environmental Contamination and Toxicology.

Vol.19.

Tucker, William. 1994. Personal communicatio": . Alabama Division

of Game and Fish, District 5,.Spannish Fore, Alabama.

U.S. EPA (United States Environmental Protection Agency). 1989.

Risk Assessment Guidance for Superfund. Vol. I. Human

Health Evaluation Manual (Part A). Office of Emergency and

Remedial Response, Washington, D.C. EPA/540/1-89/002.

U.S. EPA, 1993. Wildlife Exposure Factors Handbook Volume I of

II. Office of Health and Environmental Assessment. Office

of Research and Development. U.S. EPA. Washington, DC

20460. EPA/600/R-93/187a.

U.S. EPA, 1993a. Office of Water Guidance on Fish Sampling and

Analysis, August, 1993. Office of Water, Washington D.C.

U.S. EPA, 1994a. Ecological Risk Assessment Final Report CibaGeigy Corporation Site. Mclntosh, Alabama. U.S. EPA Office

of Emergency and Remedial Response, Environmental Response

Team, Edison New Jersey. Work Assignment No.: 0.-022.

Weston Work Order No.: 03347-040-001-0022-01. U.S. EPA

Contract No.: 68-C4-0022.

U.S. EPA, 1994b. Preliminary Ecological Risk Evaluation for LCP

Chemicals, Brunswick, GA. Memorandum. U.S. EPA Office of

Emergency and Remedial Response, Environmental Response

Team. U.S. EPA. Edison, New Jersey.

U.S. EPA. 1995. TSC 195 Criteria Chart. EPA Region IV. Water

Quality Standards Section. Water Management Division.

Atlanta, Georgia 30365.

37
U.S. EPA. 1995a. Interlabortory Quality Assurance for the

Huntsville DDT Project. Analytical Support Branch,

Environmental Services Division, Region 4, Athens Georgia.

U.S. FDA, 1992. Action Levels for Poisonous or Deleterious

Substances in Human Food and Animal Feed. Dept. Health and

Environmental Services. Public Health Service. Food and

Drug Administration. 200 C Street, S.W. Washington, DC

20204.

U.S. Fish and Wildlife Service. 1992. An evaluation of mercury

and DDT contamination in fish and sediments collected from

the Tombigbee River near Mclntosh, Alabama. July, 1992.

U.S. Fish and Wildlife Enhancement. Daphne, Alabama.

Weimeyer, S.N. and R.D. Porter. 1970. "DDE thins eggshells of

captive American kestrels." Nature. 227:737-738.

Whitr. H. 1936. "the food of kingfishers anc mergansers on the

Margaree River, Nova Scotia." J. Biol. ED. Can. 2:299-309.

(as cited in: R. Palmer, 1962).

Woodward-Clyde. 1994. Additional Ecological Studies of OU-2

Mclntosh Plant Site. Vol.1 and II. Olin Corporation/Mclntosh

Plant, Mclntosh, Alabama.

38
TABLE 1. Potential Chemicals of Concern

Associated with Superfund Sites.

Mobile River Study

Alabama

Olin/Mclntosh Site; Stauffer Sites;

Alpha-BHC Carbon Bisulfide

Arsenic Carbon Tetrachloride

Benzene Cyanide

Beryllium Mercury

Beta-BHC Thiocyanate

Cadmium Thiocarbamates including;

Chlorobenzene Molinate

Chloroform Vernolate

Copper Pebulate

4,4'-DDD Butylate

4,4'-DDE Cycloate

4,4'-DDT EPTC

Delta-BHC

Dichlorobenzene isomers

(1,2,4,5-tetrachlorobenzene,

1,2,4-trichlorobenzene)

Gamma-BHC

Hexachlorobenzene

Lead

Mercury

Pentachlorobenzene

Pentachloronitrobenzene

Tetrachloroethyene

Ciba Geiov Site;

Alpha-BHC

Ametryn

Atrazine

Beta-BHC

Bladex

Chlorobenzilate

DDD

DDE

DDT

Delta-BHC

Diazinon

Gamma-BHC

Prometryn

Simazine
Table 2. Sequence of Sampling Efforts for

the Mobile River Study.

September 1993

October 1993

November 1993

December 1993

April 1994

April 1994

November 1994

Reconnaissance of River

Sampling of sediments and water for

screening study

River Sampling for Fish Tissue

Contamination Study

River Sampling for Fish Tissue

Contamination Study

Floodplain sampling for Fish Tissue

Contamination Study

Floodplain sampling of sediment and

surface water for Screening Study

Sampling River for Fish Health

Index Study
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Table 5. Results of inorganic analyses of river

surface water samples (ug/L). Mobile River Study.

October, 1993.

STATION

STATION #

ALUMINUM

ANTIMONY

BARIUM

CALCIUM

COPPER

IRON

MAGNESIUM

MANGANESE

MERCURY*

MOLYBDENUM

POTASSIUM

SODIUM

STRONTIUM

THALLIUM

TITANIUM

ZINC

WMT-OC

80052

420

6.0U

62

18

3.3

0.53

9.3

71

2.3

3 .1

3.8

77

130

20

6.7

4.9

WMT-1C

80055

700

6.0U

68

17

5.1

0.85

4.0

76

1.7

3 .7

2.0

31

110

20U

10

9.5

WMT-2A

80067

460

6.0U

59

16

7.4

0.58

3.8

46

1.5

4 .2

1.9

26

98

20U

6.6

4.2

WMT-7C

80059

770

6.0

130

22

2.0U

0.92

5.8

77

4.9

2.5

2.6

53

180

20U

8.9

3.3

WMT-8

80072

660

6.0U

120

21

2.0U

0.80

5.3

67

1.4

2.7

2.5

35

170

20U

8.0

12

WMT-9C

80061

960

6.0U

130

21

2.1

1.1

5.6

78

3.7

3 .2

2.6

36

170

20U

12

5.3

U = Material was analyzed for but not detected. The number is

the minimum quantitation limit.

* = Ultra trace level mercury concentrations in ng/L.
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Table 7 Particle size and organic carbon content as peiccnt diy weight of

river sediments (October, 1993) and floodplain sediments (April, 1994)

Mobile River Study

medium

INORG FRAC

ORG FRACTIO

INORG FRAC

ORG FRACTIO

MT - 0 ( C ) •

INORG FRAC

ORG FRACTIO

INORG FRAC

ORG FRACTIO

INORG FRAC

ORG FRACTIO

INORG FRAC

ORG FRACTIO

INORG FRAC

ORG FRACTIO

INORG FRAC

ORG FRACTIO

INORG FRAC

ORG FRACTIO

INORG FRAC

ORG FRACTIO

INORG FRAC

ORG FRACTIO

INORG FRAC

ORG FRACTIO

gravel

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

C

0

0.

00

00

00

00

00

00

00

02

00

01

01

03

00

00

00

00

00

00

00

00

00

00

00

00

fine

gravel

0

0

0

0

1

3

0

0

0

0

0

0

0

0

2

4

0

0

0

0.

0

0

0

0

00

00

10

06

43

80

00

01

01

00

32

02

01

02

97

62

02

01

25

27

84

78

00

00

coarse

sand

87

0

3

0

1

0

20

0

5

0

3

0

21

0

2

1

0

0

2

0

1

0

0

0

91

09

46

25

18

96

94

47

81

22

61

29

78

59

03

85

38

01

78

61

96

72

00

00

medium

sand

4

0

15

0

1

0

53

0

3

10

2

0

19

0

1

0

69

0

7

1

2

0

1

0

23

01

37

40

61

66

43

59

11

38

84

26

93

49

84

99

33

21

04

00

50

61

16

12

fine very fine

sand

0

0

12

0

2

0

8

0

23

0

40

0

26

0

2

0

6

0

6

0

3

0

42

0

07

00

40

28

27

70

22

26

93

50

60

90

27

67

04

90

46

02

60

90

99

70

78

21

sard

0

0

13

0

3

0

3

0

6

0

26

0

9

0

4

1

0

0

4

0

3

0

29

0

32

01

93

49

90

67

52

16

90

32

65

87

32

38

58

44

58

01

77

65

63

59

42

22

silt

0

0

51

1

67

6

10

n

9

39

20

1

18

1

63

8

2

0

55

5

66

6

74

1

00

00

30

17

27

71

73

90

01

30

61

53

54

48

03

78

78

16

10

02

49

04

22

19

clay

6

0

0

0

7

1

0

0

0

0

1

0

0

0

4

0

0

0

13

1

9

1

0

0

77

57

58

23

56

28

34

41

46

02

25

02

40

12

28

65

03

04

16

85

69

45

28

39

totals

99

0

97.

2.

85

14

97

2

49

50

96

3

96

3.

80

19

99.

0

89

10.

89

10

97

2.

32

68

.13

.87

21

79

17

83

24

76

08

92

26

.74

77

23

.53

47

70

.30

11

89

86

14
INORG FRAC

ORG FRACTIO

INORG FRAC

ORG FRACTIO

INORG FRAC

ORG FRACTIO

medium

gravel

0.01

0.01

0.00

0.00

0.00

0.00

fine

gravel

0.08

0.06

0.00

0.00

0.00

0.00

coa

s

11

0

1

1

1.

1.

rse

and

.81

.71

.75

.09

75

27

medi um

sand

4 .57

0.32

3 .32

0.70

3.00

0.76

fine

sand

13.01

0.58

5.38

0.96

2.74

0.74

very fine

sand

37.73

1.02

4 .72

0 82

3 . 19

0 74

silt

27.69

1.80

64 .54

6.70

70.99

7.87

c:

0

0

8

1

6.

0.

lay

.16

.45

.62

.40

05

89

totals

95.04

4.96

88.33

11.67

87.72

12.28

* = Samples from floodplain survey

()** •= Replicate
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Table 20. Mean and range for DDE, DDD and DDT concentrations* in fillet and

whole body samples (mg/kg) of Largemouth Bass and Channel Catfish.

Mobile River Study. November, 1993.

Pesticide

DDE

DDD

DDT

Largemouth Bass

fillets

.035

.002 -.87

.011

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.005

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whole body

.143

.004 - 1.3

.030

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.016

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Channel Catfish

fillets

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.008

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.007

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whole body

.100

.006 - 1.1

.019

.002 - .13

.005

.002 - .021

Mean values subtended by range. MDL included as median value.
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Table 27. Fish Health Index scores for largemouth bass.

Mobile River study. October 1994.

Fish*

1

2

3

4

5

6

7

8

9

10

11

12

13

14

15

16

Mean Score

Standard

Deviation

Station Identification

MT-0

90

90

120

60

30

90

90

60

120

60

30

0

30

90

60

68.0

34.9

MT-2

90

.90

60

60

90

30

30

120

100

60

90

90

40

73.1

28.4

MT-3

60

90

30

60

30

60

90

60

90

90

30

90

150

60

60

90

71.2

30.7

MT-4

30

70

100

60

30

90

60

0

100

0

60

120

90

60

8

63.3

35.8

MT-5

90

120

60

60

30

0

60

0

60

0

60

0

0

V

41.5

39.8

MT-7

130

90

60

60

120

90

60

90

30

60

79.0

30.7

MT-8

90

90

90

60

30

120

90

60

30

60

30

68.2

30.3

MT-9

30

120

90

140

30

90

60

60

90

120

90

90

90

84.6

32.8

MT-10

120

90

90

60

90

60

60

60

30

30

90

90

90

90

90

90

76.9

24.4
Table 28. Fish Health Index Scores for channel catfish,

Mobile River study. October 1994.

Fish*

1

2

3

4

5

6

7

8

9

10

11

12

13

14

15

16

Mean Score

Standard

Deviation

Station Identification

MT-0

0

0

30

0

50

30

60

30

30

30

30

30

30

26.9

18.0

MT-2

30

30

30

30

30

30

30

30

30

30

60

30

,32.5

8.7

MT-3

30

60

60

60

60

60

60

30

60

30

60

60

52.5

13.6

MT-4

30

0

60

30

30.0

24.5

MT-5

30

30

30

30

30

30

30

30

30

30

30

30

30

60

32.1

8.0

MT-7

30

30

60

0

30

30

30

60

30

30

33.0

17.0

MT-8

30

30

30

30

30

30

30

60

30

30

30

30

30

30

30

30

31.9

<•"

7.5

MT-9

30

0

30

30

30

60

60

60

30

60

30

60

60

60

30

60

43.1

18.9

MT-10

N

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Table 32. Summary of Human Risk Level by a

finding Provided in Appendix G

Contaminant Station

DDTr

Mercury

Risk Type

carcinogenic

toxic

(non-carcinogenic)

toxic

(non-carcinogenic)

Bass

1E-5 < risk < 1E-4

risk * 1E-4

0.1 < HQ < 1.0

1.0 < HQ < 10.0

0.1 < HQ < 1.0

0.1 < HQ < 1.0

Tissue

Catfish Concentration

1E-5 < risk < 1E-4

1E-5 < risk < 1E-4

0.1 < HQ < 1.0

0.1 < HQ < 1.0

0.1 < HQ < 1.0

HQ < 1.0

mean

max

mean

max

mean

max

Above calculations based on these fillet concentrations (mg/kg)

DDTr Station 7

Mercury Station 2

average

max

average

max

0.517

1.35

0.36

0.41

0.302

0.0409

0.28

0.54
Table 33.

Hazard Quotient Calculations with Supporting

Documentation for the Wood Stork (Mycteria americana)

ingesting Mercury-contaminated prey within the 53-mile

Mobile River Study Area, 1993-94.

LIFE HISTORY INFORMATION

Home range 32 km (794,193 acres) US EPA, 1994b

Ingestion Rate 0.52 kg/day US EPA, 1994b

. Body Weight 2050 g US EPA, 1994b

PREY

Forage Fish 0.022 mg/kg Table 23

NO OBSERVABLE ADVERSE EFFECT LEVEL (NOAEL)

0.01 mg/kg body weight/day US EPA, 1994a,b

DOSE CALCULATIONS (Diet (mg/kg) X Ingestion Rate (kg/day) X Area Use Factor X I/body weight (kg))

0.022 mg/kg X 0.52 kg/day X 1 X 1/2.05 kg = 0.006 mg/kg body weight/day

HAZARD QUOTIENT CALCULATIONS (Exposure concentration/NOAEL)

0.006 mg/kg body weight/day

0.01 mg/kg body weight/day =0.60
Table 34.

Hazard Quotient Calculations with Supporting

Documentation for the Wood Stork (Mycteria americana)

ingesting DDT-contaminated prey within the 53-mile

Mobile River Study Area, 1993-94.

LIFE HISTORY INFORMATION

Home range

Ingestion Rate

Body Weight

32 km (794,193 acres)

0.52 kg/day

2050 g

US EPA, 1994b

US EPA, 1994b

US EPA, 1994b

PREY

Forage Fish 0.059 mg/kg Table 24

NO OBSERVABLE ADVERSE EFFECT LEVEL (NOAEL)

0.054 mg/kg body weight/day US EPA, 1994a,b

DOSE CALCULATIONS (Diet (mg/kg) X Ingestion Rate (kg/day) X Area Use Factor X I/body weight (kg))

0.059 mg/kg X 0.52 kg/day X 1 X 1/2.05 kg = 0.015 mg/kg body weight/day

HAZARD QUOTIENT CALCULATIONS (Exposure concentration/NOAEL)

0.015 mg/kg body weight/day

0.054 mg/kg body weight/day = 0.277
Table 35.

Hazard Quotient Calculations with Supporting

Documentation for the Belted Kingfisher (Ceryle alcyon)

ingesting Mercury-contaminated prey within the 53-mile

Mobile River Study Area, 1993-94.

LIFE HISTORY INFORMATION

Home range 8041 acres US EPA, 1994a

2.2 km shoreline US EPA, 1993

Ingestion Rate 0.198 kg/day US EPA, 1994a

Body Weight 113 g US EPA, 1994a

PREY

Forage Fish 0.022 mg/kg Table 23

NO OBSERVABLE ADVERSE EFFECT LEVEL (NOAEL)

0.01 mg/kg body weight/day US EPA, 1994a,b

DOSE CALCULATIONS (Diet (mg/kg) X Ingestion Rate (kg/day) X Area Use Factor X I/body weight (kg))

0.022 mg/kg X 0.198 kg/day X 1 X 1/0.113 kg = 0.039 mg/kg body weight/day

HAZARD QUOTIENT CALCULATIONS (Exposure concentration/NOAEL)

0.039 mg/kg body weight/day

0.01 mg/kg body weight/day =3.85
Table 36.

Hazard Quotient Calculations with Supporting

Documentation for the Belted Kingfisher (Ceryle alcyon)

ingesting DDT-contaminated prey within the 53-mile

Mobile River Study Area, 1993-94.

LIFE HISTORY INFORMATION

Home range

Ingestion Rate

Body Weight

640 acres/mi, 8041 acres

2.2 km shoreline

0.198 kg/day

113 g

US EPA, 1994a

US EPA, 1993

US EPA, 1994a

US EPA, 1994a

PREY

Forage Fish 0.059 mg/kg Table 24

NO OBSERVABLE ADVERSE EFFECT LEVEL (NOAEL)

0.054 mg/kg body weight/day US EPA, 1994a,b

DOSE CALCULATIONS (Diet (mg/kg) X Ingestion Rate (kg/day) X Area Use Factor X I/body weight (kg))

0.059 mg/kg X 0.198 kg/day X 1 X 1/0.113 kg = 0.103 mg/kg body weight/day

HAZARD QUOTIENT CALCULATIONS (Exposure concentration/NOAEL)

0.103 mg/kg body weight/day

0.054 mg/kg body weight/day =1.91
FIGURES
A

AREA OF RIVER SAMPLED

FOR FISH

COFFEEVILLE

LOCK & DAM

(RIVER MILE 116.66)

OUN AND

CIBA-GEIGY

NPL STIES

DIXIE LANDING

NEAR CHRYSLER. AL

STAUFFER

NPL SITES

RIVER MILE 26
COFFEEVILLE

LOCK & DAM

/ (RIVER MILE 116.66)

AREA OF RIVER SAMPLED

FDR FISH

OLIN AND

CIBA-GEIGY

NPL SITES

DIXIE LANDING

NEAR CHRYSLER. AL

STAUFFER

NPL SITES

RIVER MILE 26

FIGURE i

MOBILE/TOMBIGBEE RIVERS

SAMPLING STATIONS AND AREAS

MOBILE RIVER STUDY

1093-1994

SCALE IN MILES

MOBILE

BAY
FIGURE 2

Site locations for water quality and sedinent sampling in river

and floodplain wetlands, station MT-0 October, 1993 and April

1994 respectively. MOBILE RIVER STUDY
TWENTYFDUR'

MILE BEND

FIGURE 3

Site location for water quality and sedinent sampling in

the Mobile River, station MT-1, October 1993, respectively.

MOBILE RIVER STUDY
FIGURE 4

Site location for surface water and sedinent sanpling the river

station MT-2 and in the floodplain wetlands associated with Cold Creek

Swamp. October, 1993 and April, 1994 respectively. MOBILE RIVER STUDY
PUMPING STATION

FIGURE 5

Site locations for water quality and sedinent sampling

in the river (MT-7), and floodplain wetlands MT-7S, and

MT-7N, October 1993, and April 1994, respectively.

MOBILE RIVER STUDY
FIGURE 6

Site location for water quality and sediment sampling

in the river MT-9, and MT-8, and floodplain wetlands,

MT-8, October 1993 and April 1994, respectively.

MDBILE RIVER STUDY
FIGURE 7

Site locations for water quality and sediment sampling in

the floodplain wetlands, MT-11. April 1994. Tensaw Lake area

of Tensaw River. MOBILE RIVER STUDY
VHITEHDRSE

BEND*'

FIGURE 8

Sampling site locations and results for total mercury, and temperature

readings (Centigrade) in the floodplain wetlands associated with

Cold Creek Swamp, and station MT-E, April 1994. MOBILE RIVER STUDY
FLY ASH

PDND

FIGURE 9

Sampling sites and results for total mercury

in canal surface water, November, 1994. Barry Stream

Electric Generating Facilities, cooling water return to the

Mobile River. MOBILE RIVER STUDY
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Hgure 18. Range and mean concentration of Mercury in Largemouth Bass

fillet samples prepared of large fish. Mobile River Sv,dy. November, 1993

. ! I i I • I Ii I i I | I i I . i I I I

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Figure 20 Relationship between Largemouth Bass total length and whole

body concentration of total DDT. Mobile River Study. November, 1993

-0.5

150 200 250 300 350 400 450 500 550

Fish Length mm

Regression Line (Linear Fit)

Regression Output:

Constant -0.23125

StdEirofYEst 0.246132

R Squared 0.181985

No. of Observations 49

Degrees of Freedom 47

X Coefficient(s)

Std Err of Coef.

0.00125

0 000387
Figure 21. Relationship between Channel Catfish total length and whole

body concentration of total DDT. Mobile River Study. November, 1993

1.4

1.2

1

0.8

0.6

Q 0.4

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£ 0

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100 200 300 400 500

Fish Length mm

600

Regression Line (Linear Fit)

Regression Output:

Constant -0.12891

StdErrofYEst 0.183595

R Squared 0.111094

No. of Observations 42

Degrees of Freedom 40

X Coefficient(s)

Std Err of Coef.

0.00072

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Figure 26. Relationship between size of Largemouth Bass and whole body

concentration of Mercury. Mobile River Study. November, 1993

2.5

O)

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cr A 13 1

s

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150

X

200 250 300 350 400 450 500 550

Fish Length mm

Regression Line (Linear Fit)

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R Squared 0.077047

No. of Observations 49

Degrees of Freedom 47

X Coefflcient(s)

Std Err of Coef.

0.001167

0.000589
Figure 27. Relationship between size of Channel Catfish and whole body

concentration of Mercury Mobile River Study November, 1993

0.4 T

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100 200 300 400 500

Fish Length mm

600

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Constant 0.028174

Std Err of Y Est 0.060568

R Squared 0.038533

No. of Observations 40

Degrees of Freedom 38

X Coefficient(s)

Std Err of Coef.

0.000131

0.000106
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APPENDIX A

PLAN OF STUDY (POS) AND

RELATED AMENDMENT
TABLE OF CONTENTS

INTRODUCTION

Background 1

Goal 2

Analytical and Sampling Options 2

STUDY DESIGN CONSIDERATIONS 2

General Study Approach 2

Study Area 2

Potential Chemicals of Concern 3

Precision, Accuracy, Representativeness,

Completeness, and Comparability. . . 4

SAMPLING DESIGN 4

General Considerations 4

Station and Sampling Site Selection 5

Sample Analysis 6

Sample Collection and QA/QC 6

Water 7

Sediments 8

Fish for Tissue Analyses 8

Fish Health Assessment 9

DATA ANALYSIS 10

Study Area Characterization 10

Risk Assessment 10

Potentially Responsible Party 11

PROJECT SCHEDULE 12

REFERENCES 13

TABLES 15

FIGURE 24
INTRODUCTION

Background. The Mobile/Tombigbee River is a waterway

extending north from Mobile Bay to Pickwick Reservoir on the

Tennessee River Along its course upstream from Mobile Bay are

four Superfund National Priority List (NPL) Sites. The Stauffer

Chemical (LeMoyne Plant) Site located in Axis, Alabama and the

Stauffer Chemical (Cold Creek Plant) Site located in Bucks,

Alabama are approximately 20 miles north of Mobile, Alabama at

River Mile 26. The Olin Corporation (Mclntosh Plant) and the

Ciba-Geigy Corporation (Mclntosh Plant) are located in Mclntosh,

Alabama approximately 50 miles north of Mobile, Alabama at River

Mile 60 (Figure 1).

Each of these Sites have backwater or floodplain wetland

areas that are adjacent to and are hydrologically connected to

the Mobile/Tombigbee river system. From the results of

background information and on-going investigations initiated

through the CERCLA process it has been shown that each of these

wf- _ lr\d areas contaa i sediments with elevat< " levels of nazc^ lous

substances. The presence of these contaminants in the wetland

areas and the knowledge that the wetland areas are intimately

related to the Mobile/Tombigbee river system leads the U.S.

Environmental Protection Agency (EPA) to suspect that the River

system has been contaminated by the activities of these four NPL

Sites. In addition, other study findings reported by the U.S.

Fish and Wildlife Service (USFWS), Alabama Department of

Environmental Management (ADEM), and EPA have shown elevated

levels of mercury and DDT in the fish captured in this river

system. Both of these substances are known to be Chemicals of

Concern at these NPL Sites (Table 1). Information submitted to

EPA also suggests that contaminants were also directly discharged

into the river. EPA believes that the evidenced contamination

poses a potential threat to human health and the environment.

Therefore, the EPA, Region IV, Waste Management Division, Office

of Superfund, has initiated the Mobile River Study to address the

Agency's concern. In a letter dated May 25, 1993, EPA notified

the Potentially Responsible Parties (PRPs) that new operable

units had been designated for their Sites and that these operable

units would be termed the Mobile River Study. The letters

identified the Mobile River Study as operable unit 4 for the

Stauffer Chemical (Lemoyne Plant) and the Stauffer Chemical (Cold

Creek Plant), operable unit 3 for the Olin Corporation (Mclntosh

Plant), and operable unit 5 for the Ciba-Geigy Corporation

(Mclntosh Plant) The letters also notified the PRPs that EPA

was invoking its right under Section 1104(b) of CERCLA to

undertake an initial Remedial Investigation that would determine

the extent of contamination from the four NPL Sites into a 44

mile river segment which extends from just south of the Stauffer

Chemical (LeMoyne Plant) Site to jist north of the Ciba-Geigy

Corporation (Mclntosh Plant) Site "igure 1) and that EPA would

not use the special notice proviso is of CERCLA Section 122(e),

42 U S.C Section 9622(c)
Goal. The goal of the Mobile River Study is to assess the

contamination of the river, including those hazardous substances,

pollutants and contaminants associated with and potentially

emanating from the four NPL Sites and entering the designated

river segment. As a mean to this end, the objectives of this

study will include:

• Characterizing the quality of water and sediments of

the river segment and associated wetland areas in terms

of potential chemicals of concern,

• Assessing chemical contamination of selected and

representative fish species or communities of the river

segment,

• .Determining the health condition of a top predator game

fish species of the river segment.

The data collected from the initial Mobile River Remedial

Irr'p 'tigation will be used to better charac :••. r'

 7e the

ccncamination in the river segment, assess the risk to human

health and the environment, and assess contributions from the

four NPL Sites. Table 2 lists the media to be sampled and the

use of the data.

Analytical and Sampling Options. Surface water, sediment,

and fish tissue will be sampled. Tables 3 through 6 list the

data quality objectives (DQOs) for each media to be sampled. All

samples will be analyzed in accordance with analytical support

level 4. A description of the analytical support levels is

provided in Table 7 and was obtained from the Data Quality

Objectives for Remedial Response Activities Manuals (2,3) .

STUDY DESIGN CONSIDERATIONS

General Study Approach. To attain the above goal and

objectives, the river and its associated wetlands and fisheries

will be sampled at stream sites located on the reach of river

extending from the downstream vicinity of Cold Creek to an area

upstream of Mclntosh, Alabama, a distance of about 44 river miles

(Figure 1). The study will be conducted in two phases. The

first phase is a screening effort followed by a more narrowly

focused second phase of study. The sampling framework will

provide a basis for statistical comparisons between and within

station parameters. Sampling is anticipated to occur during low

flow conditions which usually coincide with the summer to early

fall period of the year.

Study Area. The study area appears fairly typical of the

Mobile/Tombigbee River in terms of general shoreline

configuration and habitat, river width, and depth. The river's
navigation channel is maintained at a minimum depth of 9 feet,

however, depths greater than 35 feet are not uncommon.

Under normal low flow conditions of the summer and early

fall, river currents are often slow enough to create a general

appearance of slack water River bank erosion appears severe and

common along the shore of the Tombigbee River upstream of the

Alabama River confluence Tidal amplitude effects of Mobile Bay

are apparent in the study area and possibly as far upstream as

Coffeeville Lock and Dam (RM 116). During low flow conditions, a

saltwater wedge originating from the Bay can extend upstream in

the Mobile River and past the confluence of Cold Creek (RM 26).

The absence of a fully mixed water column can also be expected

elsewhere in the study area because of possible thermal

stratification

The river's width varies greatly along the study area but

generally falls within a range of 500 to 1000 feet. Average

width of the river system tends to increase substantially

downstream of the confluence of the Alabama River.

Sandy sediments tend to be the predominant substrate type

essociated with the Tombigbee section of _he study area. In some

instances, the navigation channel features a scoured bottom

substrate represented by mainly an aggregate of pebbles and

coarse sand. For the Mobile River segment of the study area,

the sediment composition is typically represented by a substrate

of fine to coarse sand in the navigation channel and mud in the

bottom areas between the channel and the shoreline. Since the

Mobile/Tombigbee waterway is maintained for navigation and not

flood control, overbanking of the river during storm flow

conditions is not unusual. With overbanking, nutrients and

settleable solids are supplied throughout the river floodplain

and associated wetlands. Much of the organic and inorganic

materials, which could include the potential chemicals of

concern, flooding the wetlands are deposited as sediment and are

subjected to a myriad of possible chemical and biological

processes. In these processes, materials, whether natural or

man-made, can be transformed from their original chemical form,

sequestered in the floodplain sediments, or mobilized for

reprocessing and transport elsewhere.

Potential Chemicals of Concern. Potential chemicals of

concern have been identified from the findings of investigations

on the wetland areas associated with the four Superfund Sites

(Table 1). Mercury and DDT are of particular concern because of

their reported concentrations in the sediments and aquatic biota

sampled in the designated operative units of the Superfund Sites.

Environmental concerns for these contaminants are heightened due

to the collective findings of other studies reported by the

Alabama Department of Environmental Management, EPA, and the

USFWS. The compound DDT was found in the tissues of Largemouth

Bass collected from the river in the vicinity of Mclntosh,

Alabama Concentrations for DDT fron whole fish analyses ranged
from 0.36 to 34.52 ppm and were indicated as excessive for fisheazing bird species (1). Over the pass two decades, EPA and ADEM

have reported mercury concentrations exceeding 1 ppm in filets

from Lc'.rgemouth Bass collected from the reach of the river

currently the subject of this study. A mercury level of 1 ppm in

fish filets exceeds the Food and Drug Administration's action

level for considering fish consumption advisories.

In preparing the list of concern, the assumption was made

that all of the cited constituents (Table 1) could potentially

reach the river and pose an environmental concern. Collectively,

40 chemicals of potential concern were identified. Many of these

compounds, if released to the rJ.ver, would potentially escape

measurement for various reasons including dilution effects,

volatility, hydrolysis, or minimal analytical detection limits of

the chemical methods used for their analysis.

Precision, Accuracy, Representativeness, Completeness, and

Comparability. The precision, comparability and accuracy of

sample analyses are addressed in the Analytical Support Branch

Operations and Quality Control Manual (4) or as specified by the

existing US-EPA standard procedures and protocols for contract

analytical laboratory programs. All samples will be collected

and handled in accordance with the Environmental Compliance

Branch Standard Operating Procedures and Quality Assurance Manual

(5) and/or the Ecological Support Branch Standard Operating

Procedures Manual, 1992 (6). This will result in collection of

representative samples of the various media. Completeness will

be achieved for 99 percent of all the samples collected (1

percent may be lost as a result of sample breakage in the

laboratory or during transport). It is also anticipated that 99

percent of the samples analyzed will result in valid data. Fish

collection and preparation of fish tissue samples for chemical

analyses will be accomplished with guidance provided by in USEPA

Fish Sampling and Analysis: A Guidance Document for Issuing Fish

Advisories (Draft) and Fish Field and Laboratory Methods for

Evaluating the Biological Integrity of Surface Water (7,8). The

protocol developed by the Tennessee Valley Authority (TVA) (9)

will be followed for the assessment of fish health conditions in

the study area of the river. All samples collected for chemical

analysis will be booked into the ESD or Contract Laboratory

Program (CLP) laboratories through the Regional Sample Control

Center (RSCC), to ensure availability of space and continuity of

data.

SAMPLING DESIGN

General Considerations. To narrow the array of potential

chemical constituents of concern as listed in Table 1, a select

number of representative sediment and water samples from the

study area will be analytically screened for the elements and

compounds of interest. Chemicals quantified in the screening

phase of the study will then be targeted in the second or
definitive phase of the study when sediments, water, and fish

samples will be collected at the eleven stations indicated in

Figure 1. Based upon findings from the definitive study, a

follow-up or third phase of the study could be formulated, if

• necessary, to address data gaps relative to the initial study

objectives and respond to additional assessment questions of

concern.

Summer through early fall is the anticipated period of

sampling for this study. Generally during this time, low flow

conditions are encountered. Low flows tend to minimize dilution

effects, provide for a more stable sampling environment and pose

worst, case seasonal conditions for water quality and aquatic

biota. Minimum dissolved oxygen concentrations and maximum water

temperatures for example are generally encountered during this

time and can pose a natural stress to the aquatic life. These

conditions when coupled with other environmental stressors, such

•as the potential chemicals of concern, can add to the natural

adverse stresses with the effects being manifested in the health

condition of the exposed fish community.

Over the course of the spring through early fall periods of

the year, fish will experience their maximum growth rate and

develop their greatest body fat or lipid content by winter.

Chlorinated pesticides such as DDT are extremely lipid soluble;

hence, the late fall to winter would be the preferred time to

sample fish tissue for lipid soluble pesticides. High flow

conditions are not, however, suited for the planned sampling

regime. The fish tend to be more dispersed and their habitat

more difficult to access under high flow conditions. For these

reasons, the summer to early fall season of the year was selected

as the sampling period for the study.

Station and Sampling Site Selection. The genera] location

of sampling stations selected for this study are indicated in

Figure 1. Eleven sampling stations are planned for this study.

Of this number three stations will serve as reference sites (MT8,

MT9 and MT10) and will be strategically located outside the

potential influence of the four Superfund Sites. The station

array does not purposely reflect randomness. Instead, station

placement is biased to characterize upstream/downstream

conditions relative to the sources of potential chemicals of

concern. A similar characterization is also extended to the

Alabama River and its confluence with Mobile/Tombigbee River

(Figure 1) .

Station selections as indicated in Figure 1 will remain

tentative until the screening phase of the study is completed.

In the screening phase, each of the indicated river stations will

be surveyed for the primary type of bottom substrate

characterizing the river and the presence and type of floodplain

forested wetland flanking the station area. The bottom substrate

of both the river and wetlands woiId likely represent areas for

accumulating fine particles and sorbed chemicals. Depending upon

the survey findings of the screening study, the tentative
stations may be adjusted longitudinally upstream or downstream to

help assure that these types of substrates, wetlands in

particular, are represented in the sampling strategies. In

evaluating sampling results via an upstream/downstream

comparison, similar substrate types are essential to the

evaluation. Likewise, the reference stations will be selected to

also facilitate comparison of similar substrates. Once the final

stations are determined, they will be geographically referenced

with a Global Positioning System (GPS).

To characterize the quality of the surface water and

bottom sediments at each station, sampling locations will be

keyed to a transect extending across the river and into adjacent

floodplain wetlands. For the river, per se, the transect will be

divided into three segments of similar lengths. Facing upstream,

the left, middle, and right segment of the transect will be

designated the west(A), mid(C), and east(B) sampling zones for

the river. Similarly, the floodplain wetlands associated with

the transect, sampling locations will be designated either the

west or east bank and subject to the same sampling strategy

applied to the river.

In sampling the fish communities, a su --area of the river

around each designated station will be defined as the fish

sampling area. Fish collected from anywhere within this area

will be referenced to the designated station. Depending on the

level of effort required to collect the required fish species and

sample size, the up and downstream boundaries of the subarea will

probably be different from station to station. In any case, the

upstream and downstream boundaries of each subarea will also be

georeferenced.

Sample Analysis. All sediment and surface water samples

will be analyzed in the laboratory for extractable organic

compounds (base/neutral and acid extractables - including routine

pesticides and PCBs), volatile organic compounds, metals, special

pesticides and cyanide (Table 1). Additional laboratory analysis

required on only sediments include total organic carbon, bulk

density, and moisture content. For water, additional laboratory

analyses include dissolved organic carbon, particulate organic

carbon, and ultra trace levels for mercury. Fish tissue samples

will be analyzed for routine pesticides, extractable organic

compounds and metals including mercury.

Sample Collection and QA/QC. For both the screening and

definite phase of the study, all samples will be collected in

accordance with the appropriate Standard Operating Procedures

(SOPs). The quality assurance and quality control procedures

described in this section will help insure that representative

samples are collected from the various media sampled. Table 8

lists the number of samples anticipated for each media sampled,

including QA/QC samples. The reported sample counts for water

and bottom sediments are considered to be a maximum possible

number for the study. As later discussed in the water sampling
section, actual sample numbers will largely depend on river

conditions at the time of the survey.

Duplicate or split samples will be collected at two of the

sample locations for each matrix and analyzed for extractable

organic compounds and metals. This will provide a check for

sampling techniques. In addition, each week water samples are

collected, water trip blanks will be prepared with organic-free

water for volatile organic analysis. The trip blanks will be

handled and stored with the samples collected from the site.

This will provide a check to determine if samples may have been

contaminated during handling, storage, and shipment.

Preservative blanks will be collected at the beginning of the

project and at the end of each week water samples are collected

to insure the integrity of the preservatives. Equipment rinse

blanks will be collected from equipment cleaned on-site, using

organic/inorganic free water, at least once a week or more

•frequently at the discretion of the field project manager and

analyzed for parameters of concern. The equipment rinse blanks

will be collected to ensure that the sampling equipment is

properly field cleaned.

All sample containers will be obtain .c f urn the BSD bottle

repository. Sample containers used will be in accordance with

the requirements specified in Appendix A of the ECBSOPQAM, and/or

the Ecological Support Branch Standard Operating Procedures

Manual (6) Table 9 indicates types of analysis required, type

preservatives needed, and kinds of sample containers to be used

in the study.

Water. At each of the river stations the left, middle, and

right zones of the river transect will be sampled. The sampling

will begin with establishing a vertical profile of the water

column in terms of temperature (C°), dissolved oxygen (ppm), pH,

conductivity (umhos/cm), and REDOX (millivolts). For this task,

remotely deployed, calibrated probes will be required. The

profiles will provide a means to assess the degree of water

column stratification and determine the need for two level

sampling of the water column. To define stratification, it is

assumed that a surface to bottom gradient will usually occur for

these particular parameters during the given sampling period,

i.e., dissolved oxygen and temperature values decreasing with

depth.

For purposes of this study, the boundary dividing the upper

and lower strata of the water column can be defined by the depth

at which the dissolved oxygen concentration decreases to one ppm

or less or when the REDOX readings are less than a positive 200

millivolts. These values will help determine the apparent

oxidizing or strongly reducing stratums of the wate:: column which

are significant factors affecting the chemical quality of the

water. Another factor affecting the chemical characteristics of

the water column is density stratification. Determining the

presence and location of a thermocline or the

freshwater/saltwater interface (clinograde) serves to identify
the upper and lower stratums of the water column based upon

density differences. For this study, the thermocline is defined

as the water column depth at which temperature changes one degree

C° or more within a one meter depth interval. To identify the

clinograde boundary, an increase in conductivity to 1000 umhos/cm

or more indicates the freshwater/ saltwater interface of the

water column.

For water sampling sites where the water column is fully

mixed, a water grab sample will be collected from mid depth.

Under a stratified condition, the sample will be obtained at the

mid point of depth within each of the two stratified layers of

water. In cases where the sampling site in the floodplain

wetland is covered with water, surface grab samples will be

obtained prior to the collection of the substrate samples. All

samples will be kept in designated containers, labeled, treated

with an appropriate preservative, stored and shipped to a

designated laboratory for analyses. The maximum surface water

sample count for the definitive phase study (Table 8) is based on

the assumption that at all stations, the water column is

stratified in the three sampling zones and that the associated

f."oc "plain wetlands are also inundated on b^th sides of the

river. Under this assumption, a total of twelve water samples

per station is anticipated.

Sediments. Surficial samples (top 10 to 15cm) of river or

wetland sediments will be generally collected along the same

transect line used for surface water. Sediment sampling will be

accomplished with either a coring or dredging device. Three

sediment replicate samples per sand and/or mud substrate found at

each station are required. Similarly, three replicate bottom

substrate samples will be collected from the selected wetland

sites. When taking a sample, minimum disturbance to the

sediments is required to help assure representativeness of the

site. Each sample will be visually examined on site, its

characteristics noted, and then processed. All samples will be

labeled, preserver1, stored, and shipped to a designated

laboratory for analyses. Sediment samples will be subject to

particle size determinations and analyses for the potential

chemicals of concern. Total sediment sample count is based on

the assumption that each station will feature two substrate types

in the river and that wetlands flank both sides of the river.

This assumption would provide for the collection of 12 sediment

samples per station.

Fish For Tissue Analyses. The sampling of the fish

communities in the study area may involve a wide array of capture

gear including fish shockers, trotlines, and nets because of the

species and large numbers of fish required for tissue analyses.

The goal of the sampling effort is two fold. First, collect two

species of gamefish in sufficient numbers to facilitate a fish

tissue contamination assessment relative to three size groups per

species. Largemouth Bass (a top predator) and Channel or Blue

8
Catfish (bottom feeder) are the gamefish species of interest.

Secondly, collect sufficient numbers of a species representing

the forage fish community of the river. Guidance for the

collection, field dressing, and processing of fish tissue samples

for chemical analysis is provided in USEPA Fish Sampling and

Analysis: A Guidance Document for Issuing Fish Advisories

(Draft) February 1993 and Fish, Field and Laboratory Methods for

Evaluating the Biological Integrity of Surface Waters (7,8).

The field processing of the sampled fish will begin with

assessing the general external condition, total length, and wet

weight of each specimen collected for analyses. Large, medium,

and small size specimens for each species will be aggregated into

respective groups of similar lengths. The size interval for

aggregating the samples will be determined in the field. The

control for each size group will be established by the total

length of the smallest member of the aggregate measuring no less

•than 25 percent the total length of the largest member. Total

length will serve as the standard measurement of size. In

summary, each fish species will be represented by three size

groups.

From each size group, six replicate composite samples of

filets will be prepared. A composite will be comprised of five

filets with each filet from a different fish. Three of these

composite samples will be analyzed for metals and three for

organic compounds. The analyses of filets will serve mainly to

aid in providing the basis for public health risk assessments.

For an ecological risk assessment, whole body analyses are

required. To facilitate this analyses, five whole fish specimens

will be collected for each species. The five specimens will be

selected to represent a range of sizes. Each whole fish group

will be analyzed for body concentrations of metals, extractable

organic compounds, and pesticides.

A third species or an assemblage of species will be

collected to represent the forage or prey component of the

river's fishery. Typically, this component of the fish community

is made up of minnows and other small fish species which are

preyed upon by a variety of fish eating animals. Hence, the

forage community can serve as a critical pathway for the

bioaccumulation of mercury and pesticides in higher trophic

levels of the riverine ecosystem.

Depending on abundance, three replicate composite samples of

a forage species or an assemblage of several forage species will

be collected. Each composite should be comprised of a minimum of

200 grams of fish weight. Each composite will be analyzed for

the same chemical constituents assigned the filet whole fish

assessment. Fish sample container, analysis required, and

preservation needs are given in Table 9.

Fish Health Assessment. This component of the study will

involve the development of a Fish Health Assessment Index (FHAI)

which is an autopsy-based survey that quantifies the effects of
environmental stressors on resident fish species. The fish

species of interest in this assessment is the Largemouth Bass

which is considered to be a primary top predator of the

Mobile/Tombigbee River system. The FHAI will be the results of

on-site autopsies of freshly collected specimens of the targeted

species. Specimens for the autopsy work will be submitted to the

field laboratory with blind identity as to sample station

location. Autopsy parameters and methods fol.lowed will be from

the protocol developed by TVA (9). This assessment will not

serve to elucidate cause and effects but rather to identify

stress gradients in the study area and their geographical

association with the Superfund sites.

DATA ANALYSIS

Study Area Characterization. To characterize the study

area, the sampling framework was organized to facilitate a

compare and contrast design. In this approach, eleven river

stations were identified for sampling. Of this total, three and

eight locations were designated as reference and potential impact

sites, respectively. The reference stations were all positioned

outside the influences from the NPL sites; hence, they are

assumed to be outside the effects of potential chemicals of

concern. Each station will be characterized in terms of its

prominent physical, chemical, and selected biological features.

Analytical results will be statistically treated to detect

significant differences between and within stations. Data will be

displayed in the form of tables, figures, and maps to determine

the general health and condition of selected components of the

ecosystem.

Risk Assessment. A human health and ecological risk

assessment will be performed on the data obtained in this study.

The human health assessment will follow the guidelines of the

Risk Assessment Guidance for Superfund Volume 1 Human Health

Evaluation Manual (Part A) (10). For ecological risk assessment,

a "weight of evidence" approach will be followed. Pertinent

guidance will be derived from Risk Assessment Guidance for

Superfund Volume II Environmental Evaluation Manual Interim Final

(11) ; Framework for Ecological Risk Assessment (12) ; Peer

Review Workshop Report on a Framework for Ecological Risk

Assessment (13); and Report on the Ecological Risk Assessment

Guidelines Strategic Planning Workshop (14). Other documents and

reference material that may be considered in the course of the

ecological risk can include the following:

• Exposure Analysis Modeling System: User Guide for EXAMS

II Version 2.94. (15).

• The Potential for Biological Effects of Sediments -

Sorbed Contaminants Tested in the National Status And

Trend Program. (16).

10
Primary Responsible Party. By the examination of the

results showing the distribution of contaminants in the

sediments, water, and fish, EPA will assess the contribution of

contaminants from each of the four NPL Sites to the river area of

concern. Also the findings of the risk assessment will be used to

determine the threat to human health and the environment from the

contaminants and their distribution. The distribution of

contaminants, risk to humans, and risk to the ecosystem will

serve as a basis to aid in the determination of any necessary

further actions to be taken in the study area of the river.

Further actions under CERCLA may be required. If no further

action is required at the Sites, a "No Action" Record of Decision

(ROD) will be established.

11
PROJECT SCHEDULE

Plan of Study Review

Meeting to Discuss Review Comments

Revision to Plan of Study

9/7-20/93

9/15/93

9/25/93

Phase I

Phase II -

Screening Sampling

Screening Result

Fish Sampling

Fish Ti isue Results

10/4-8/93

12/15/93

11/1-10/93

5/15/94

Phase II - Sediment/Water Sampling

- Analytical Results

7/15/94

9/15/94

Phase II - Fish Health Assessment

Assessment Results

8/1/94

9/15/94

Draft Final Report Review

Final Report

11/1/94

12/15/94

12
REFERENCES

1. United States Fish and Wildlife Service. An Evaluation of

Mercury and DDT Contaminants in Fish and Sediments Collected

From the Tombigbee River near Mclntosh, Alabama. U.S. Fish

and Wildlife Enhancement, Daphne, Alabama, July 1992.

2. United States Environmental Protection Agency. Data Quality

Objectives for Remedial Response Activities, Volume 1,

Development Process. Office of Emergency and Remedial

Response and Office of Waste Programs Enforcement,

Washington, DC 20460. EPA 540/G-87/003A, March 1987.

3. United States Environmental Protection Agency. Data Quality

Objectives for Remedial Response Activities. Example

Scenario. Office of Emergency and Remedial Response and

Office of Waste Programs Enforcement, Washington DC 20460.

EPA/540/G-87/004, March 1987.

4. United States Environmental Protection Agency. Analytical

Support Branch Operations and Quality Control Manual

(ASBOQCM). Region IV, Environmental Services Division,

Athens, Georgia 30605, September 1990.

5. United States Environmental Protection Agency, Region IV,

Environmental Services Division. Environmental Compliance

Branch Standard Operating Procedures and Quality Assurance

Manual. February 1991.

6. United States Environmental Protection Agency. Ecological

Support Branch Standard Operating Procedures Manual. Region

IV, Athens, Georgia 30605, April 1989.

7. United States Environmental Protection Agency, Region IV.

Fish Sampling and Analysis: A Guidance Document for Issuing

Fish Advisories (Draft). February 1993.

8. Klemm, Donald J., Quentin J. Stober, and James M. Lazorchak.

Fish Field and Laboratory Methods for Evaluating the

Biological Integrity of Surface Waters. U.S. Environmental

Protection Agency, Environmental Monitoring Systems

Laboratory, Cincinnati Office of Modeling, Monitoring

Systems, and Quality Assurance, Office of Research and

Development, Cincinnati, Ohio 45268. EPA/600/R-92/111,

March 1993.

9. Adams, Marshall S., Allen M. Brown, and Ronald W. Goede. A

Quantitative Health Assessment Index for Rapid Evaluation of

Fish Condition in the Field. Transactions of The American

Fisheries Society. Volume 122 (1), January 1993.

10. United States Environmental Protection Agency. Risk

13
Assessment Guidance for Superfund Volume I Human Health

Evaluation Manual (Part A) Interim Final. U.S.

Environmental Protection Agency, Office of Emergency and

Remedial Response, Washington, DC 20450. EPA/540/1-89/002,

December 1989.

11. United States Environmental Protection Agency. Risk

Assessment Guidance for Superfund Volume II Environmental

Evaluation Manual Interim Final. Office of Emergency and

Remedial Response, Washington DC 20460. EPA/540/1-89/1001,

March 1989.

12. United States Environmental Protection Agency. Framework

for Ecological Risk Assessment. Risk Assessment Forum.

Washington DC 20460. EPA/630/R-92/001, February 1992.

13. United States Environmental Protection Agency. Peer Review

Workshop Report on a Framework for Ecological Risk

Assessment. Risk Assessment Forum. Washington DC 20460.

EPA/625/3-91/022, February 1992.

14. United States Environmental Protection Agency. Report on

the Ecological Risk Assessment Guideline Strategic Planning

Workshop. Risk Assessment Forum. Washington DC 20460.

EPA/650/R-92/002, February 1992.

15. Burns, L. A., 1989. Exposure Analysis Modeling System:

User Guide for EXAMS II Version 2.94. United States

Environmental Protection Agency, Athens, Georgia, 30605.

16. Long, Edward R. and Lee G. Morgan. The Potential for

Biological Effects of Sediment-Sorbed Contaminants in its

Natural Status and Trends Program. National Oceanic an

Atmospheric Administration, National Ocean Service, NOAA

Technical Memorandum NOS DMA 52. Seattle, Washington, 1989.

14
TABLE 1. Potential Chemicals of Concern

Associated with Superfund Sites.

Mobile River Study

Alabama

Ol in/Me Intosb. Site; Stauffer Sites:

Alpha-BHC Carbon Disulfide

Arsenic Carbon Tetrachloride

Benzene Cyanide

Beryllium Mercury

Beta-BHC Thiocyanate*

Cadmium Thiocarbamates including;

Chlorobenzene Molinate*

Chloroform Vernolate*

Copper Pebulate*

4,4'-DDD Butylate*

4,4'-DDE Cycloate*

4,4'-DDT EPTC*

Delta-BHC

Dichlorobenzene isomers

(1,2,4,5-tetrachlorobenzene*,

1,2,4-trichlorobenzene)

Gamma-BHC

Hexachlorobenzene

Lead

Mercury

Pentachlorobenzene*

Pentachloronitrobenzene*

Tetrachloroethyene

Ciba Geiqv Site;

Alpha-BHC

Ametryn*

Atrazine*

Beta-BHC

Bladex*

Chlorobenzilate*

DDD

DDE

DDT

Delta-BHC

Diazinon*

Gamma-BHC

Prometryn*

Simazine*

* Special analyical services needed: not included with TAL/TAL

15
TABLE 2. Data Uses

Mobile River Study

Alabama

MEDIA RIVER RISK

SAMPLED CHARACTERIZATION ASSESSMENT

PRP

DETERMINATION

SURFACE WATER

SEDIMENT

FISH TISSUE

FISH HEALTH

YES

YES

YES

YES

YES

YES

YES

YES

YES

YES

YES

YES

16
TABLE 3. Data Quality Objectives

Surface Water Samples

Mobile River Study

Alabama

Media: River and wetland surface water

Location: Mobile/Tombigbee River and tributaries

Data Types: A. In situ - pH, specific conductivity,

temperature, dissolved oxygen, redox.

B. Laboratory analyses - volatile organic

compounds, base/neutral/acid extractable organic

compounds, (including routine pesticides and PCB's)

metals, ultra trace levels of total mercury,

special pesticides (Table 1) , cyanide, total

dissolved organic carbon, and total part^culate

organic carbon.

Sample Type: Environmental, grab sample

Xn Situ Analytical Data Level: Level 2

Laboratory Analytical Data Level: Level 4

Field Quality Control Samples;

5 percent duplicate/split samples, (VOA trip blank,

metals and cyanide preservative blank, and

equipment rinse blank collected once per week or as

needed).

Sampling Procedures:

See section 4, Environmental Compliance Branch

Standard Operating Procedures and Quality Assurance

Manual and Ecological Support Branch Standard

Operating Procedures Manual (5,6).

17
TABLE 4. Data Quality Objectives

Sediment Samples

Mobile River Study

Alabama

Media: Sediment

Location: Mobile/Tombigbee River and its tributaries

Data Types: A. In situ - visual inspection

B. Laboratory Analyses - Volatile organic

compounds, base/neutral/acid extractable organic

compounds, (including routine pesticides and PCB's)

metals, special pesticides, cyanide, total organic

carbon and bulk density (Table 1).

Sampling Type: Environmental, grab sample

Jn Situ Analytical Data Level: Not applicable

Laboratory Analytical Data Level: Level 4

Field Quality Control Samples:

5 percent duplicate or split samples

Sampling Procedures:

See section 4, Environmental Compliance Branch

Standard Operating Procedures and Quality Assurance

Manual and Ecological Support Branch Standard

Operating Procedures Manual (5,6).

18
TABLE 5. Data Quality Objectives

Fish Tissue Samples

Mobile River Study

Alabama

Media: Fish tissue

Location: Mobile/Tombigbee River and its tributaries

(see figure 1)

Data Types: A. In situ - none

B. Laboratory analyses: - Base/neutral/acid

extractable organic compounds (including

routine pesticides and PCB's), metals

(excluding cyanide), and special pesticides

(excluding Thiocarbamates).

Sampling Type: Environmental

Laboratory Analytical Data Level: 4

Field Quality Control Samples; 5 percent duplicate/split samples

Sampling Procedures:

See the Fish Sampling and Analysis: A Guidance Document for

Issuing Fish Advisories and Fish Field and Laboratory Methods

for Evaluating the Biological Integrity of Surface Waters

(7,8) .

19
TABLE 6. Data Quality Objectives

Fish Health Assessment

Mobile River Study

Alabama

Media: Fish

Location: Mobile/Tombigbee River and tributaries (see

figure 6)

Data Types: A. In situ - none

B. Laboratory analysis - visual examination

of fish body and internal organs

Sampling Type: Environmental

Laboratory Analytical Data Level: 2, 3

Field Quality Control: None

Sampling Procedure: TVA Protocol for Conducting Fish Health

Assessments (9).

20
TABLE 7. Analytical Levels

Mobile River Study

Alabama

Level Description

Field screening. This level is characterized by the use

of portable instruments which can provide real-time data

to assist in the optimization of sampling point locations

and health and safety support. Data can be generated

regarding the presence or absence of certain contaminants

at sampling locations.

Field analysis. This level is characterized by the use

of portable analytical instruments which can be used onsite, or in a mobile laboratory stationed near a site.

Depending upon the types of contaminants, sample matrix,

and personnel skills, qualitative and quantitative data

can be obtained.

Laboratory analysis using methods other than the contract

laboratory (CLP) or EPA, Region IV, BSD routine

analytical services. This level is used primarily in

support of field studies using standard EPA approved

procedures. Some procedures may be equivalent to CLP/ESD

routine analytical services, without the requirements for

rigorous QA/QC documentation.

CLP or BSD routine analytical services. This level is

characterized by rigorous QA/QC protocols and

documentation and provides qualitative and quantitative

analytical data.

Non-standard methods. Analyses which may require method

modification and/or development. CLP special analytical

services are considered level 5.

21
TABLE 8. Sample Collection Summary

Mobile River Study

Alabama

SAMPLE DESCRIPTION

TRIP BLANKS (WATER)

TRIP BLANKS (SEDIMENT)

PRESERVATIVE BLANKS

EQUIPMENT RINSE BLANKS

MATRIX WATER (EXT.)

MATRIX WATER (METALS)

MATRIX SEDIMENT (EXT.)

MATRIX SEDIMENT (METALS)

WATER SAMPLES

SEDIMENT SAMPLES

FISH TISSUE

TOTAL

SCREENING

PHASE NUMBER

SAMPLES

0

0

0

0

30

27

57

DEFINITIVE

PHASE NUMBER

SAMPLES

132

114

350

596

QA/QC

DEFINITIVE

PHASE

4

4

4 (PER WEEK)

1 (1 PER WK IF

CLEANED)

2

2

2

2

13 (DUPLICATES

& SPLITS)

14 (DUPLICATES

& SPLITS)

35 (DUPLICATES

& SPLITS)

83

22
TABLE 9 Sample Containers and Preservatives

Mobile/Tombigbee River Study

Alabama

WATER ANALYSIS

Extractao'e Organ c

(includirg rout^e c°s:icices

and DCB s;

Speci^c

 Desticides

Volatile Organic

Metals

Ultra Trace Level Mercury

Cyanide

Total Dissolved Organic

Carbon

Total Participate Organic Carbon

SEDIMENT SAMPLE ANALYSIS

Extractable Organic (includes

routine pesticides/PCB s

Specific Pesticides

Metals/Cyanide

Volatil e Organic

Total Organic Carbon

Bulk Density and Percent

Moisture

iuer ameer glass Teflcn liner

1 - 4 liter amber glass (may

need as many as 3 4 lite r

arnbers) with Teflon liner

3 40ml glass vial s (going to

ESD)

2 40 ml glass vial s (going to

CLP)

1 - 1 polyethylene with

polyethylene lined closure

1 - 500 ml Teflon container

with Teflon liner

1 - 1 liter polyethylene with

polyethylene lined closure

1 - 500 mL polyethylene with

polyethylene lined cap

1 - 500 mL polyethylene with

polyethylene lined cap

1 - 8 oz widemouth glass jar

with Teflon liner

1 - 8 oz widemouth glass jar

with Teflon liner (may need as

many as 3 - 8 oz glass)

1 8 oz widemouth glass jar

with Teflon liner

1 - 4 oz widemouth glass jar

with Teflon liner

1 8 oz widemouth glass jar

with Teflon lined li d

1 - qt widemouth glass jar

Iced 4° C

Iced 4° C

4 drops 1+1 HCL iced to 4° C

4 drops 1+1 HCL iced to 4° C

50$ HN03 to pH <2. iced 4° C

2 mis non-metal grade HCL

analyzed before use

NaOH to pH > 12 iced 4° C

Filtered (0 45fj) and acid

preserved

Non filtered and acid preserved

Iced 4° C

Iced 4° C

Iced 4° C

Iced 4° C

Iced 4° C

Iced 4° C

FISH SAMPLE ANALYSIS

Routine Pesticides

Metals

Each whole body wrapped in foil

(shiny side out) and then placed

in plastic bag and sealed

Filet composite wil l be

aggregated and placed together

on same foil and wrapped

together then sealed in plastic

bag

Each whole body placed HI

y ast.c oag nd sealed Filet

composites wi 11 be aggregated in

the sare p'astic bag and sealed

Iced 4° C freeze ASAP or

within 24 hours

Iced 4° C freeze ASAP or

within 24 hours

23
Mobile/Tombigbee River

Study Plan Amendment

February 28, 1994

INTRODUCTION: The Environmental Protection Agency initiated the

Mobile/Tombigbee River Study in the fall of 1993 to address the

agency's concern that hazardous material originating from four NPL

sites may be contaminating the river and posing a potential threat

to human health and the environment. Implementation of the study

was according to a Plan of Study which provides the bases for the

investigation, identifies the goals and objectives, lists the

chemicals of concern, and reports study design considerations and

sampling strategies.

In brief the study employs a two phase approach to sampling. Phase

I involved a screening study in which sampling was conducted at

selected river stations located upstream and downstream of the NPL

oi..-^s. The sampling effort was desxgned to facilitate a

characterization of the chemical quality of water and sediments of

the river and associated floodplain wetlands. Sampling for the

screening phase of the study was partly completed the week of

October 4, 1993. Sampling of the floodplain wetlands was not

possible at this time for lack of permission for land access to the

property; however, river water and sediments were obtained.

Following the field work of the screening study, the second phase

of the investgation was initiated with the collection of fish

samples for tissue analyses. The fisheries work began the week of

November 1, 1993 and was completed approximately six weeks latter.

The analyese of the tissue samples are scheduled for completion in

April, 1994 .

Analytical results for the chemical screening of water and sediment

samples from the river show that chemicals of concern were found at

concenrations comparable to reference conditions or- below minimun

detection limits for the analytical procedures employed. These

findings give rise to Lue question that possibly the floodplain

wetlands may play a more significant role than does the river in

the fish tissue contamination issue.

The basis for this question involves several considerations. All

four of the NPL sites are located in floodplain wetlands. With

winter river stages, overbanking and the inundation of the

floodplain wetlands are a common occurance in the Mobile/Tombigbee

River system. As overbanking occurs, fish can migrate from the

river to the floodplain where they can merge with the wetland food

chain. Duration of inundation in the wetlands can span several

weeks thus providing both opportunity and pathways for fish to.

forage for food which may be contaminated with a chemical of

concern and/or be directly exposed to contaminants in the wetlands.

Potentially, contaminants in the floodplain wetlands could be

directly link to the NPL sites, derived via the river from upstream
sources, or directly deposited in the wetlands through air

deposition. In any case, the testing of surface water, sediments,

and fish tissue samples from the floodplain wetlands would be

necessary to adequately address the assessment question relative to

the wetlands. To proceed with this sampling, the following

sampling stratigy is provided.

SAMPLING APPROACH: Sampling of the floodplain wetlands during

times of inundation is planned for the period of March/Aril, 1993.

River stages are anticipated to be sufficiently elevated during

this time to effect overbanking and flooding of the wetlands.

Under these hydrologic conditions, access to the floodplain

wetlands from the river is presumed to be possible by boat.

Sampling of the floodplain wetlands will be approached in the same

manner as the Phase I screening study. Samples from the floodplain

wetlands associated with the NPL sites and selected reference

•wetland will be analytically screened for chemical constituents of

concern.

The sampling effort for the floodplain wetlands will involve the

collection of sediments, surface water, and fish samples. Samples

will be analyzed for the suite of chemicals identified in the

initial Plan of Study. This sampling will remain within the scope

of the initial Plan of Study and the stated study goals,objectives,

sampling and analytcial methods, and quality assurance procedures.

Sample parameter will remain basically the same as initially

planned with the exception of adding fish stomach content analyses.

STATION AND SAMPLING SITES: For purpose of the chemical screening

phase of the study, five floodplain wetland areas were selected for

sampling. Four of these coincide with river stations MT-0, MT-2,

MT-7, and MT-8. The former three stations are located within the

potential influence of the NPL sites. Station MT-8 is a backwater

reference location which is positioned upstream ofthe NPL sites and

outside their influence. A second backwater wetland reference

station in the Tensaw Lake area is being researched for use as the

fifth station(MTL).

Within each of the floodplain wetlands identified, sampling site

selection will deviate from the initial Plan of Study. Originally

sample collection in the floodplain wetlands would have occurred at

three locations along an extension of a cross sectional transect of

the river. Instead, each of the three sampling sites will be

scattered to locations most representative of the floodplain

wetland station. The positioning of these sites will be

accomplished with the benefit of reconnaissance prior to sampling.

SITE SAMPLING: Sediment, surface water, and fish remain the

primary sampling media of inter?st. Because of the environmental

setting of the floodplain wetla .ds during periods of inundation,

some deviation will be required : i the sampling methods reported in

the initial Plan of Study. The. 3 changes ire following.

Water: The surface water c lumn is e -pected to be vertically
mixed in the floodplain wetlands during the overbanking period.

Under a non-stratified condition, the water column will be sampled

at mid depth at each of three sites. Sample collection, handling,

and preservation will be according to the initial Plan of Study.

Sample summary is provided in Table I.

Sediments: Since sampling is anticipated to occur from a

boat, a remote operated coring or augering device will be used to

sample the sediments. Three replicate samples will be gathered at

each sampling site, composited, and subsampled for chemical

analyses. Handling and preservation of samples will be according

to the initial Plan of Study. Sample summary is provided in Table

I.

Fish: The Largemouth Bass, Channel Catfish, and forage fish

are the species of interest. An array of sampling gear will

probably be required to catch representative samples of the target

species for each of the floodplain wetland areas selected for

study. For the Bass and Catfish, the goal of the sampling is to

collect adequate numbers of fish to constitute a sample of

sufficient filet and whole body biomass to re-present the three size

group established in the Plan of Study. Similarly, the forage fish

community will be sampled until sufficient numbers are available to

constitute an adequate composite of biomass for chemical analyses.

The stomach content of the captured fish will be examined to

determine the type of food items consumed relative to size class.

The food items will also be analyzed for chemical constituents of

concern poviding the biomass of the sample is of adequte size. A

sample summary is provided in Table I.
APPENDIX B

IN SITU MEASUREMENTS AND RESULTS
In Situ Measurements and Results

DO concentrations were near levels of saturation and generally

similar among the stations except MT-8, a back water area of the

river known as Three Rivers Lake. DO values were slightly less

than saturation and characterized by a profile of decreasing

concentrations from surface to bottom. The lower concentration of

DO near the bottom probably reflected decreased effects of vertical

mixing in the water column as compared to the river.

Thermal stratification of the water column was evident at

station MT-2 and was related to effects of the heated discharge

ori^ ..nating with the Barry steam electric generation facility of

the Alabama Power Corporation (Figure 4) . The facility's permitted

Point of Discharged (POD) to the river was located approximately

500 yards upstream of station MT-2. Surface water temperature was

35.4 C which decreased to 28.1 C near the river bottom. The

thermocline occurred at a depth of about 10 feet. Water

temperatures at stations MT-1 and MT-0 ranged between 27.8 and 29.0

C and were sirrd lar to the readings measured below the thermocline

at station MT-2. Temperature profiles of the water column in the

upper reach of the study area (MT-7, 8, and 9) were characterized

by cooler temperatures ranging from 24.5 to 26.4 C.

Conductivity reading indicated that the river was fairly well

mixed along the vertical plain of the water column, except in the

case of the navigation channel at station MT-0 (Table 2). At this

station, the conductivity profile for the navigation channel

indicated a relatively narrow range in values (305 to 311 umhos)

from the surface to a depth of about 35 feet where the conductivity

increased to 750 umhos and then to 6250 umhos at the bottom. The

increased conductivity could best be explained by the probable

effects of a strata of denser, saline water extending up channel

from Mobile Bay, a result of freshwater stream flow merging with an

estuary.

Another aspect of the conductivi,.\ regime at station MT-0 was
its profile established outside of the channel area at site MT-OC.

This shallower area which flanked the navigation channel and

adjoined the eastern shore of the river featured a relatively

narrow range of readings from surface to bottom. These values were

two fold greater in magnitude than readings in the river channel

site (Table 2). This contrast would indicate that the water of the

navigational channel was not fully mixed cross sectionally at

station MT-0 at least during the time of sampling. The increased

conductivity along the east side of the river at this location was

likely reflecting residual effects of previous salinity intrusion

into this area of the study reach. Upstream from station MT-0, the

conductivity profiles for stations MT-1 and MT-2 were slightly

decreased from the observation at the downstream sampling sites.

In the upper reaches of the study area n'-_.r Mclntcsh, Alaocuna,

the remaining stations for the screening study were located

(stations MT-7, MT-8, and MT-9). The former station was situated

approximately 300 yards downstream from the Olin Corporation's

river loading facilities (Figure 5) . At this site the conductivity

profiles for the navigation channel and the shallower, near shore

region flanking the east bank of the river shared a similar range

of readings (Table 2) . The river at this point appeared both

vertically and horizontally mixed. The conductivity regime at this

location was about 30 percent greater than observed at stations MT7, MT-8 and MT-9 located upstream of the Mclntosh area and probably

best reflected a source of more mineralized water inflowing to the

river.

The pH profiles for all stations sampled followed a fairly

narrow range of 7.1 to 7.7.

The above reported profiles of the in situ parameters were

considered in selecting sites and location in the water column to

obtain representative samples for water analyses. Other

considerations involved in the water and sediment sampling strategy

included safety relative to anchoring the sampling boat, barge

traffic and currents. In Table 2, sampling depths and sites are

indicated.
Table B-l. Vertical water quality profiles.

Mobile River Study. October, 1993

STATION

MT-OC

MT-OB

MT-1A

DATE

10-6-93

10-7-93

10-7-93

DEPTH

(FT)

1

4

7*

10

13

15.5**

1

6

11

16

21

26

31

35

38**

1

6

11

16

21

26

31

36

41

46

51

56**

TEMP.

(°C)

28 .5

28.4

28.5

28 .5

28.5

28.5

28.0

27. 9

27. 9

27. 9

27.8

27.8

27.8

27.8

28.1

29 .0

29.0

29. 0

29.0

29. 0

29.0

29.0

29.0

29.0

29.0

29.0

29 . 0

D.O.

(mg/L)

6.5

6.4

6.3

6.3

6.3

6 .3

6. 9

7.0

7.0

6 . 9

6.9

6.4

6.6

6.4

4.5

7.0

7.0

6.9

6.8

6 .8

6.8

6.8

6.8

6.8

6.'7

6.7

6.7

COND.

(^mho)

630

690

718

713

762

757

305

307

309

309

310

311

309

750

6250

284

284

283

282

282

282

283

285

281

285

282

283

pH

7.2

7.2

7.1

7.2

7.2

7.2

7.2

7.2

7.2

7 .2

7.2

7.2

7.2

7.1

6.5

7.2

7.3

7.2

7.2

7.2

7.2

7.2

7.2

7.2

7.2

7.2

7.2
Table B-l cont.

STATION

MT-1B

MT-2A

MT-7C

MT-7B

MT-9C

MT-8

DATE

10-6-93

10-8-93

10-7-93

10-7-93

10-7-93

10-7-93

DEPTH

(FT)

1

4

7

10*

13

16

19

20

1

4

7

10

13*

17

20**

1

4*

7**

1

6

11

16

21**

1

4*

7

10**

1*

3

6**

TEMP.

(°C)

28. 9

29.0

28.9

29.0

29.0

28 . 9

28 .6

28. 6

35.4

35.2

32 .5

30 .8

28.8

28.3

28 .1

26.3

26 . 1

21.0

26 .2

26.2

26 .2

26 .2

26 .2

26.4

26 .3

26 .3

26.3

25.6

25 .5

24 . 5

D.O.

(mg/L)

7.1

6.9

6.8

6.8

6. 8

6.7

6 .5

6.5

6 .8

6.7

6.7

6.6

6.6

6.6

6.5

8.9

8.3

8.0

8.2

8 .2

8.2

8.1

8.0

8.9

8.9

8.8

8.8

7.5

7.1

4 . 9

COND.

(/umho)

287

287

286

285

285

288

295

298

249

247

243

239

233

230

229

410

490

528

467

480

480

476

473

354

354

354

354

309

306

235

pH

7.3

7.3

7.3

7.2

7.3

7.2

7.2

7.2

7.2

7.2

7.2

7.2

7.2

7.2

7.2

7.2

7.3

7 .3

7. 5

7. 5

7. 5

7.6

7.5

7.7

7.7

7 .7

7.7

7.2

7.2

7. 1

Station - A,B,C respectively indicate

quarter points of the river cross sect

* - depth of water column gr^t sample

** - sediment grab sample fo 'h^rn; _;, l

lefc, center and right

jon facing upstream.

f"r chemical analyses.

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APPENDIX C

FISH COLLECTION PROTOCOL,

MOBILE RIVER STUDY,

1993-94.
Fish Collection Protocol, Mobile River Study, 1993-94.

Largemouth Bass, Channel Catfish and forage fish for the Mobile

River Study were captured between November, 1993 and 1994. All

collected species were harvested using boat mounted

electrofishing equipment. Additionally Channel Catfish were also

collected in cheese-baited box traps. Once caught, the fish were

immediately transferred to a cooler which held bagged ice and was

lined with plastic. After a short reach of the river was

electrofished, the fish were sorted with gloved hands, measured,

placed in individual plastic bags with station number and

specific fish number noted, sealed, returned to cooler and

information recorded onto a field log sheet. At the end of the

fishing session, fish were transported to the processing area.

FILLETING PROCESS

Talc free gloves were worn anytime fish were handled and

were changed between every composite or individual fish sample.

Individual fish were grouped according to size class and species.

Individual fish were measured to determine total body length

(from the tip of the nose to the tip of the compressed caudal

fin) in millimeters and weighed to the nearest gram while still

in their individual bags, and this information was recorded on

the log sheet. The fish were then removed from the plastic bag

with gloved hands and placed in a glass pan with other fish of

the same composite sample. The bag was weighed and its weight

recorded on the log sheet. When all five fish had been weighed

and measured, five to ten scales were removed from the left side

of each of the fish below the lateral line and posterior to the

pectoral fin for possible fish aging studies. These scales were

placed in appropriately marked individual manila envelopes. Two

tags were prepared for each fish with its individual fish number

and the addition of an "R" on one tag and an "L" on the other to

identify the right or left fillet and placed in the mouth of the

fish, (Example 1-32R, 1-32L). The fish were then scaled

(Largemouth Bass) or skinned (Channel Catfish), rinsed with

organic free water and placed in a clean glass pan. The

individual fish were transferred to a glass plate for filleting.

The right and left fillet, including the belly flap, were removed

using stainless steel fillet knives. Sex was determined whenever

possible and recorded. External anomalies were noted and

recorded. The left fillet was weighed and placed on a piece of

plastic (with the numbered tag from the fish's mouth) for metals

analysis and the right fillet weighed and placed on a piece of

aluminum foil, with its numbered tag, for organics analysis. The

left fillets for each composite sample were individually wrapped

in plastic and placed in a single plastic bag. The right fillets

were individually wrapped in aluminum foil and placed in a single

plastic bag. Custody tags were placed with the fillets,

information logged and the fish frozen for transport to the

Athens Ecological Support Branch Lab for processing.
PROCESSING FILLET SAMPLES

The five fish fillets comprising each composite were removed

from the freezer to partially thaw so that they could be cut into

small pieces with a stainless steel knife. These pieces were

placed in a blender which had been prechilled with dry ice and

blended to a uniform consistency. Additional dry ice was added

as necessary to enhance this process. The thoroughly blended

sample of all five fillets was then placed on foil or in a

plastic bag (depending on analysis protocol), loosely sealed and

allowed to sublime in the freezer for approximately 48 hours.

Fish were removed from the freezer, thoroughly mixed and aliquots

measured for analysis of metals or organics. The aliquots were

then tagged and shipped on dry ice to the analytical

laboratories. Chain-of-custody protocol was followed throughout

the study.

WHOLE BODY FISH

Whole body fish were tagged and logged and placed in the

freezer at the field processing laboratory upon arrival from the

field. The frozen fish were transported to Athens for final

processing. Processing consisted of thawing the fish enough to

allow them to be cut into small pieces, placed in a grinder and

ground. The intestinal tracts of all catfish were removed

because of the presence of cheese bait which was used to bait the

traps. The fish were sent through the grinder three times to

facilitate thorough mixing of all parts of the fish. Aliquots

were measured for metals and organic analyses, wrapped, frozen,

chain-of custody initiated, and shipped on dry ice to analytical

laboratories.

DECONTAMINATION PROCEDURES.

All equipment, with the exception of dip nets, were cleaned

with tap water and laboratory detergent using a brush, if

necessary, to remove particulate matter and surface films. The

equipment was rinsed thoroughly with tap water, rinsed thoroughly

with deionized water, rinsed twice with isoproranol, rinsed

thoroughly with organic free water and allowed to air dry as long

as possible or wrapped in aluminum foil for longer storage. All

equipment was decontaminated after each sample.

Dip nets were cleaned with laboratory detergent and

thoroughly rinsed with tap water, allowed to air dry as long as

possible and wrapped in plastic bags. Dip nets were changed

between stations.
APPENDIX D

FISH HEALTH INDEX (FHI) PROTOCOL
SOP XXXI

Revision No. 2

Dale: Decerrber 30,1994

Page 1 of 12

Fish Health Index (FHI) Protocol

Introduction

This protocol is designed to assess the general health of fish populations in a reservoir or river system.

The THI is based on the Health Assessment Index (HAI) developed by Adams et. al (1993) and the Fish

Health / Condition Assessment Procedures developed by Goede (1993). Autopsies are performed on a

population of live fish, concentrating on the following specific tissues and organs. The tissue and organs

examined include:

Length Fat

Weight Gall bladder

Eyes Hind gut

Gills Kidney

Pseudobranch Liver

Thymus Spleen

Fins Blood

Opercles

Each tissue or organ is assigned a numerical value based on general condition or degree of damage. Each

fish receives a total score by summing individual values for each tissue or organ. The mean score for

a "population" offish can be compared statistically with mean scores of other populations.

A. Apparatus and Equipment

Hematocrit Centrifuge

Hematocrit tubes

Hematocrit seal

Hematocrit rack

Protometer

Forceps

Blunt nose scissors

Scalpels

Fillet knives

Glass pans

Balance

Measuring board

Latex rubber gloves

MS-222 (tricaine methanesulfonate)

Camera

Camera stand

Film

Paper towels

Kimwipes

Trash bags
SOP XXXI

Revision No. 2

Dale: Decerriber 30,1994

Page 2 of 12

Opti-visors

Stereo microscope

Field Collection Equipment

Electro fish ing unit

Generator

Dip nets

Live well

Eelectricians gloves

Rubber boots

Boat

ri. Fish Collection

The optimum sample size is 15 fish, however, as few as 10 is acceptable. Fish should be collected via

electrofishing or a passive collection method. Active collection methods such as seining should be

avoided as they can produce external injuries that may bias results. Upon collection, immediately place

fish in a live well and disturb them as little as possible until they are anesthetized in preparation for the

autopsies. A minimum length of 250 mm is recommended for bass and 325 mm for catfish, although

it is possible to autopsy smaller fish.

C. Autopsy Procedure

Immobilize fish by placing them in a 100 mg/1 (ppm) solution of MS-222 (tricaine methanesulfonate).

A 100 ppm solution is made by adding 4 g powdered MS-222 to approximately 10 gallons (40 liters) of

water. Wait 5 to 10 minutes for the anesthetic to take effect. Fish are properly anesthetized when

regular, rhythmic opercular activity ceases. At this point fish will remain alive but anesthetized for

several hours.

External Examination

Begin the external exam by laying an anesthetized fish on a measuring board and recording its length in

mm. Next transfer the fish to a scale and record its weight in grams. Now lay out the fish and examine,

in order, the following tissues and organs, recording the condition of each tissue or organ with a number

or letter code as listed below:

Normal (N) - No aberrations in evidence. Good "clear" eye.

Exopthalmic (popeye) (El, E2) - Swollen, protruding eye. More commonly referred to as

"popeye". It is coded as El or E2. This refers to the presence of exopthalmia in one eye or two

eyes.
SOP XXXI

Revision No. 2

Dale: December 30,1994

Page 3 of 12

Hemorrhagic (HI, H2) - Refers to bleeding in the eye.

"Blind" (Bl, B2) - This is a very graphic category and you need not know whether the eye is

functionally blind. It generally refers to opaque eyes. The nature of the opacity is not important

here.

"Missing" (MI, M2) - An eye is actually missing from the fish.

"Other" (OT) - Any manifestations which do not "fit" the above. Describe it in the

remarks column.

Fins;

It must be remembered that this particular assessment system is concerned primarily with health and

condition. It is not concerned with aesthetic values. Eroded or "ragged" fins are definitely indicative

or a departure from normal condition and health. Previously ciousd -,ns which are completely healed

over and showing no evidence of the active erosion are, for the purposes of this assessment, considered

normal. Th^ evaluation of fins is relative to the degree of active erosion process in evidence. For the

purposes of this procedure, the number and location of fins involved is not significant. If only one fin

is displaying active erosion, the observation must be ranked and recorded. If several fins are displaying

erosion with unequal severity, the observation must refer to the most severe in evidence. This unequal

nature of the observations, in this case, is less significant in a full 15 fish sample. The classification is

as follows:

No active erosion (0) - Normal appearing fins with no active erosion. This would include

previously eroded fins which were completely healed over.

Mild active erosion (1) - Active erosion process but no hemorrhage or secondary infection

in evidence.

Severe active erosion (2) - Active erosion with hemorrhage and/or secondary infection

in evidence.

Note! Make a general remark relative to which fins were involved and any other observation of

special significance. There is a space for this type of entry at the bottom of the data collection

worksheet. This is particularly important in the summary.

Opercles:

It is necessary only to observe the degree of shortening of the opercles. The classification is as follows:

Normal opercle (0) - No shortening; gills completely covered

Mild shortening (1) - Slight shortening of the opercle with a very small portion of the gill

exposed
SOP XXXI

Revision No. 2

Date: December 30,1994

Page 4 of 12

Severe shortening (2) - Severe shortening of the opercles with a considerable portion of the gill

exposed.

Gills:

Normal (N) - No apparent aberrations in gills. Be very careful in this observation. The gill

can easily be effected by the manner in which the fish is handed during and after collecting. Be

alert and aware.

"Frayed" (F) - This generally refers to actual erosion of tips of gill lamellae resulting in "ragged"

appearing gills. Mere separation of gill lamellae can be construed to be "frayed" but that

condition may have been caused by something as simple as the manner in which the gill was

exposed by the investigator.

"Clubbed" (C) - This refers to swelling of the tips of the gill lamellae. They can often appear

bulbous or "club" like. The causes are not pertinent until interpretation is considered.

"Marginate" (M) - A graphic description of a gill with a light discolored margin along the distal

ends or tips of the lamellae or filaments. Margination can be and often is associated with

"clubbing". If both seem to apply it is not a problem. It is important that you noted that it was

not normal. Use the one which seems most appropriate.

"Pale" (P) - This refers to gills which are definitely very light in color. Severe anemia_ can

result in gills which are discolored to the point of being white. Severe bleeding induced during

sampling of blood can also result in somewhat pale gills. Gills begin to pale somewhat after

death also. This is not uncommon in fish taken from nets. All of this should be considered in

making the observation.

Other (OT) - Any observation which does not fit above. Describe in remarks.

Pseudobranchs:

Normal (N) - The pseudobranch is located dorsally and anterior to the gills in the branchial

cavity and can be easily observed under the operculum. The normal pseudobranch is quite "flat"

or even concave in aspect and displays no aberrations.

Swollen (S) - The "swollen" pseudobranch is convex in aspect and not difficult to discern upon

close examination.

Lithic (L) - Mineral deposits in pseudobranchs, manifested by appearance of white, somewhat

amorphous spots or foci.

Swollen and Lithic (S&L) - Lithic pseudobranchs are often also swollen.
SOP XXXI

Revision No. 2

Dale: December 30,1994

Page 5 of 12

Inflamed (I) - This is a generic use of the term, inflamed, and would more appropriately be

termed "redness" because it also includes observations of hemorrhage and any other cause of

redness. The term, "inflamed" has been traditionally used to describe this condition and is thus

continued for that reason.

Other (OT) - This term will cover any manifestation observed in the pseudobranch which is

not covered in the categories. Be sure to describe in remarks.

Thymus;

Assessment of the thymus involves degree of petechial or "pinpoint" hemorrhage.

No Hemorrhage (0) - The thymus displaying no hemorrhage is considered to be a normal

condition, although this assumption is still under investigation. Caution must be exercised here

because when the thymus involutes or ceases to function there is no observable petechial

hemorrhage. This happens normally as the fish mature. In salmonids, involution of the thymus

is thought to happen at two or three years of age but there is considerable disagreement among

investigators about that point.

Mild hemorrhage (1) - A few red spots or petechial hemorrhages in evidence. This might

be only two or three small spots.

Moderate hemorrhage (2) - More than a few red spots.

Severe Hemorrhage (3) - Many "pin point" hemorrhages in evidence with some of them

coalescing. The general area may also have a swollen tumescent appearance but that should be

recorded in remarks.

Internal Examination

Once the external examination has been completed, position the fish "belly up" and, using sharp/blunt

scissors, make a ventral cut from the anal vent forward to the pectoral girdle. Do not insert the scissors

so far that the internal organs are damaged. In largemouth bass and other centrarchids carefully cut

through the pectoral girdle to lay open the pericardial cavity and expose the heart and aorta. In catfish

leave the pectoral girdle intact.

Liver;

Upon opening the fish, immediately inspect and record the color and condition of the liver. This must

be done prior to collecting blood. Collecting blood and even minor hemmorhaging sustained during

dissection can quickly drain color from the liver.
SOP XXXI

Revision No. 2

Dale: December 30,1994

Page 6 of 12

Normal (A). Good, solid red color.

Lighter-or less vivid red color. Not so pale as to be classed general discoloration. Still

considered to be normal.

"Fatty" liver (C). Light tan color, such as "coffee with cream".

Nodules (NO) Nodules in the liver, i.e. white mycobacterial cysts and incipient nodules, such

as those in hepatoma.

Focal discoloration (DF). Change of color in local areas or foci of the liver.

General Discoloration (DG). Color change in the whole liver.

Other (OT). Aberration or deviation in liver which does not fit into above scheme. Class as

OT and describe in remarks.

Blood

Following inspection of the liver, collect blood. Blood is collected with a sharpened, heparinized

microhematocrit tube via a cardiac puncture. In bass blood is tapped from the aorta. By inserting a

finger into the buccal cavity an gently pressing upwards, the heart and aorta can b^ i.amooilized against

the roof of the pericardial cavity, making cardiac puncture much easier. In catfish the same technique

can be used to immobilize the heart and aorta, but, since the pericardial cavity is not open, the cardiac

puncture is more difficult. Once the heart and aorta are immobilized between the floor and raised roof

of the pericardial cavity, the sharpened capillary tube is thrust through the membrane separating the

pericardial and coelomic cavities and into the pericardial cavity in the suspected location of the aorta.

If the aorta is punctured, a spurt of dark red blood will partially fill the tube. If a clear yellow or pale

red fluid enters the tube, the aorta was missed. Try again with a fresh hematocrit tube. When the aorta

has been punctured, blood can be pumped into the tube by gently releasing and then reapplying pressure

on the aorta with the finger still inserted in the buccal cavity. The tube once filled is sealed with

hematocrit clay and placed in a numbered rack. Make every effort to ensure samples are matched with

the appropriate fish. After collecting a blood sample, proceed with the autopsy as outlined below.

Note: After 30 minutes or the collection of 6 to 8 samples, whichever comes first, place tubes in a

hematocrit centrifuge and spin for five minutes. Hematocrits should be read within 2 hours of collection

or before the blood begins to coagulate. To read hematocrits:

Obtain an hematocrit reader and place the filled hematocrit tube so that the bottom of the red

portion is at the 0 line and the meniscus of the clear piasma is at the 100% percent mark. The

packed red cell volume of the blood is the hematocrit and is read as a percentage of the total

blood volume.
SOP XXXI

Revision No. 2

Dale: December 30,1994

Page 7 of 12

The leucocrit is the buffy of gray zone which consist of the leucocytes or white blood cells an

is also read as percent of the total volume on the hematocrit reader.

The protein content of the plasma is determined using a protein refractometer. Take the

hematocrit tube and break it off just above the buffy, gray zone to obtain the clear plasma and

express the plasma onto the clear glass surface of the protein refractometer. Avoid placing any

glass fragments from the tube on the refractometer. Read the weight/volume percent of protein

in the refractometer. The protein refractometer should be calibrated prior to use with distilled

water.

After collecting a blood sample, proceed with the autopsy by examining, in order, the following tissues

and organs, recording the condition of each tissue and organ with a number or letter code listed below:

..iesenteric Fat;

0 - No fat deposited around the pyloric caeca.

1 - Slight, where less than 50% of each cecum is covered with fat.

2 - 50% of each cecum is covered with fat.

3 - More than each cecum is covered with fat.

4 - Pyloric caeca are completely covered by a large amount of fat.

Spleen;

Black (B) - The "black" is actually a very dark red color of the spleen.

(Red coloration of the spleen. There is subjective variation among investigators

as to whether the spleen is black or red, but both condition are considered

normal.)

Granular (G) - Granular or "rough" appearance of the spleen. This is also considered normal.

Nodular (NO) - The spleen contains or manifests fistulas or nodules of varying sizes. These are

often cysts, such as those encountered with mycobacterial infections.

Enlarged (E) - Spleens can, on occasion, be significantly and noticeably enlarged.

Other (OT) - Occasionally there are observable, gross aberrations which do no fit the above.

There may be spleens with a gray mottling and some with very small spleens. These should be

classed as "other" and described in remarks.

Hindiiut:
SOP XXXI

Revision No 2

Date: December 30,1994

PageS of 12

A short distance of the hind gut should be "opened". This should, in fact, have been accomplished as

mentioned above when the body cavity was incised. If not, it must be opened to expose the "inner

lining" or mucosa. Using the handle of a forceps or some other appropriate blunt instrument, lightly

"scrape" out the contents of the hindgut so that you can observe relative reddening or inflammation.

0 - No inflammation or reddening of the hindgut.

1 - Mild or slight inflammation or reddening of the hindgut

2 - Moderate inflammation of the hindgut.

3 - Considerable, severe inflammation or reddening of the hind gut.

KHney.

Normal (N) - Good firm dark red color lying relatively flat dorsally in the visceral cavity

along the length of the ventral surface of the vertebral column. It will be necessary to pull

the swimbladder and some of the mesentery aside to expose the kidney to view.

Swollen (S) - Enlarged or swollen wholly or in part.

Mottled (M) - Gray discoloration, mottled or "patchy" in appearance ranging from scattered

patches of gray to total gray discoloration. This is not to be mistaken with the superficial

gray appearance induced by the mesenteric membranes on the surface of the kidney. This

should be moved aside before observation is recorded.

Nodular (NO) - The kidney may have a "granular" or nodular appearance and texture. This

may be induced by granulomatous concretions.

Urolithiasis (U) - This condition is also known as nephrocalcinosis and involve mineral

material in the deposition of a white or "cream-colored" amorphous tubules of the kidney, it

can range in appearance from very small white spots to severe involvement with very large

"serpentine" deposits. These sites of deposition are not to be confused with the Stannius

bodies corpora of Stannius which are present in salmonid kidneys and have endocrine

function. The Stannius bodies are generally not associated with the tubules and usually occur

at the "edges" in an area about midway along the kidney. They appear more globular than do

the urolithic deposits.

Other (OT) - This is used to class any aberrations which do not fit into the above scheme.

Record it as OT and describe it in the remarks.

Gall Bladder -

0 - Yellow, empty of partially full

1 - Yellow, full or distended
SOP XXXI

Revision No. 2

Date: December 30,1994

Page 9 of 12

2 - Grass green

3 - Dark green

Sex:

Male (M) - Testes observed

Female (r) - Ovaries observed Note if "ripe" with eggs

Unknown (U) - immature or undeveloped gonads

General Observations and Remarks

Anything which appears to be abnormal should be noted. It is recommended that the mesenteric

tissues in the visceral cavity be checked for hemorrhage and inflammation and if these conditions are

present they should be so noted in general remarks.
SOP XXXI

Revision No. 2

Date: December 30,1994

Page 10 of 12

D. Scoring

Once autopsies are completed, transcribe the number and letter codes assigned to the condition of

each tissue/organ to a numerical score according to the table below. Note that no score is given to

sex, fat deposits, and gall bladder. Conditions associated with these tissues/organs are not considered

pathological but simply a physiological or nutritional state that varies on a seasonal or other periodic

basis.

Tissue or Organ Condition

N

E1.E2

H1.H2

B!,B2

M1.M2

OT

0

1

2

Score

0

30

30

30

30

30

0

10

20

Tissue or Organ

Spleen

Hind Gut

Condition

B

GR

NO

E

OT

0

1

2

3

Score

0

0

30

30

30

0

10

20

30

Opercles

Gill

Pseudobranchs

0

1

2

N

F

C

M

P

OT

N

S

L

SL

I

OT

0

1

2

3

0

10

20

0

30

30

30

30

30

0

30

30

30

30

30

0

10

20

30

Liver

Blood:

Hematocrit

Leucocrit

Plasma Protein

N

S

M

NO

U

OT

A

C

NO

DF

DG

OT

<4

>6

<3

0

30

30

30

30

30

0

30

30

30

30

30

0

30

0

30

0

30

30
SOP XXXI

Revision No. 2

Date: December 30,1994

Page 11 of 12

E. Calculations and Summary Data

The following calculations are useful in assessing the overall fish health. The specific calculations

performed will be determined by the specific objectives of a given study. Many of the following

calculations can be done by hand, but for most studies the use of a computer or at least pocket

calculator with a function key for calculating mean and standard deviation is recommended.

Ktl:

The value for Ktl is a ratio of length to weight. It is routinely expressed as Ktl x IO5 to

mitigate the problem of carrying so many decimal places in the records. Use the following

equation to obtain a value:

W x 10*

Ktl = L3

Where:

W = Weight in grams

L = Total length in millimeters

Mean and Standard Deviation:

The mean is determined by totaling all of the values for the observations and dividing by the

number of the observations. In-depth discussion of the standard deviation is beyond the scope

of this presentation. Again, a pocket calculator or computer is recommended for making this

calculation. In each sample of fish examined calculate means and standard deviations for the

following observations:

Length

Weight

Ktl

Hematocrit

Leucocrit

Plasma protein

These calculations are made on raw data from autopsy sheets, not from assigned scores.

Fish Health Index (FHF) Score;

Calculate an FHI score for each fish by totaling the numerical score for each tissue/organ.

For a given sample or population of fish calculate the mean and standard deviation from the

individual FHI scores for fish in that sample. The mean FHI scores between samples can be

compared statistically.
SOP XXXI

Revision No. 2

Dale: December 30,1994

Page 12 of 12

Values as Percents of Total Sample

This calculation expresses as a percent the number of fish in a sample with a specific

physical/pathological condition. As an example, consider the eyes. The number of fish with

normal eyes in a given sample is divided by the total number of fish in the sample (x 100) to

yield the percentage of normal-eyed fish in that sample. Similarly, the percentage of fish in a

sample with one blind eye (Bl) can be calculated. A percentage can be calculated for any or

all physical/pathological conditions. It is particularly useful for pinpointing a high incidence

of disease in any one particular tissue or organ.

F. Interpretation

The state-of-the-art in this area has not been refined. Normal and abnormal conditions for each tissue

and organ have been fairly well defined, but measures of overall fish health based on FHI scores, for

r

 ample, are still being evaluated. The interpretation of overall fu,h health is complicated by the

varying sensitivity of different fish species to environmental stresses. A viable strategy for

investigating potential hazardous waste sites is to statistically compare the mean FHI score for a fish

sample from a suspected "impacted" site with the mean score for fish from an established or selected

"unimpacted" reference or background site on a species to species basis. Other strategies may prove

equally enlightening as more FHI studies are completed.
APPENDIX D

FISH HEALTH INDEX (FHI) PROTOCOL
Length:

Weight;

Goede (1993)

AUTOPSY CLASSIFICATION KEY

Total length in millimeters

Weight in grams

Ktl:

Sex;

Eyes;

Fins;

Opercles;

Gills;

Pseudobranch:

Thvmus;

Fat:

F

M

W X 10s

L

3

F, note if ripe

EXTERNAL EXAMINATION

Normal (N)

Exopthalmic (popeye) (E1,E2)

Hemorrhagic (H1,H2)

Blind (B1,B2)

Missing (M1,M2)

Other (OT)

No active erosion; previous erosion healed over (0)

Mild erosion with no bleeding (1)

Severe active erosion with hemorrhage and/or

secondary infection (2)

No shortening (0)

Mild shortening (1)

Severe shortening (2)

Normal (N)

Frayed (F)

Marginate (M)

Clubbed (C)

Pale (P) Other(OT)

Normal (N)

Swollen (S) Lithic (L)

Swollen and Lithic (SL)

Inflammed (I) Other (OT)

No hemorrhage (0)

Mild hemorrhage (1)

Severe hemorrhage (2)

INTERNAL EXAMINATION

None

Slight, < 50% of each cecum is covered

50% of each cecum is covered

>50% of each cecum is covered

100% of each cecum is covered

(0)

(1)

(2)

(3)

(4)
Spleen: Black (B) Red (R)

Granular (G)

Nodular (NO)

Enlarged (E)

Other (OT)

Hindcrut: No inflammation (0)

Slight inflammation (1)

Severe inflammation (2)

Kidneys; Normal (N)

Swollen (S) Mottled (M)

Granular (G) Urolithic (U)

Other (OT)

Liver; Red (A) Light red (B)

Coffee & Cream (C)

Nodular (D)

Discoloration, focal (E)

Discoloration, general (F)

Other (OT)

Gallbaldder; Yellow, empty or partially full (0)

Yellow, full or distended (1)

Grass green (2)

Dark green (3)

Blood:

Hematocrit Volume of red blood cells (erythrocytes) expressed

as percent of total blood volume. Centrifuged 5

minutes.

Leucocrit Volume of white blood cells (leucocytes) expressed

as percent of total blood volume. "Buffy" zone of

the packed cell column.

Plasma protein Amount of protein plasma, expressed as gram percent

(grams per 100 ml).
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SOP XXXI

Revision No. 2 '

Date: Decen*er 30,1994

Page 1 of 12

Fish Health Index (FHI) Protocol

Introduction

This protocol is designed to assess the general health of fish populations in a reservoir or river system.

The FHI is based on the Health Assessment Index (HAI) developed by Adams et. al (1993) and the Fish

Health / Condition Assessment Procedures developed by Goede (1993). Autopsies are performed on a

population of live fish, concentrating on the following specific tissues and organs. The tissue and organs

examined include:

Length Fat

Weight Gall bladder

Eyes Hind gut

Gills Kidney

Pseudobranch Liver

Thymus Spleen

Fins Blood

Opercles

Each tissue or organ is assigned a numerical value based on general condition or degree of damage. Each

fish receives a total score by summing individual values for each tissue or organ. The mean score for

a "population" of fish can be compared statistically with mean scores of other populations.

A. Apparatus and Equipment

Hematocrit Centrifuge

Hematocrit tubes

Hematocrit seal

Hematocrit rack

Protometer

Forceps

Blunt nose scissors

Scalpels

Fillet knives

Glass pans

Balance

Measuring board

Latex rubber gloves

MS-222 (tricaine methanesulfonate)

Camera

Camera stand

Film

Paper towels

Kimwipes

Trash bags
SOP XXXI

Revision No. 2

Dale: December 30,1994

Page 2 of 12

Opti-visors

Stereo microscope

Field Collection Equipment

Electrofishing unit

Generator

Dip nets

Live well

Eelectricians gloves

Rubber boots

Boat

B. Fish Collection

The optimum sample size is 15 fish, however, as few as 10 is acceptable. Fish should be collected via

electrofishing or a passive collection method. Active collection methods such as seining should be

avoided as they can produce external injuries that may bias results. Upon collection, immediately place

fish in a live well and disturb them as little as possible until they are anesthetized in preparation for the

autopsies. A minimum length of 250 mm is recommended for bass and 325 mm for catfish, although

it is possible to autopsy smaller fish.

C. Autopsy Procedure

Immobilize fish by placing them in a 100 mg/1 (ppm) solution of MS-222 (tricaine methanesulfonate).

A 100 ppm solution is made by adding 4 g powdered MS-222 to approximately 10 gallons (40 liters) of

water. Wait 5 to 10 minutes for the anesthetic to take effect. Fish are properly anesthetized when

regular, rhythmic opercular activity ceases. At this point fish will remain alive but anesthetized for

several hours.

External Examination

Begin the external exam by laying an anesthetized fish on a measuring board and recording its length in

mm. Next transfer the fish to a scale and record its weight in grams. Now lay out the fish and examine,

in order, the following tissues and organs, recording the condition of each tissue or organ with a number

or letter code as listed below:

Eyes;

Normal (N) - No aberrations in evidence. Good "clear" eye.

Exopthalmic (popeye) (El, E2) - Swollen, protruding eye. More commonly referred to as

"popeye". It is coded as El or E2. This refers to the presence of exopthalmia in one eye or two

eyes.
SOP XXXI

Revision No. 2

Date: December 30,1994

Page 3 of 12

Hemorrhagic (HI, H2) - Refers to bleeding in the eye.

"Blind" (Bl, B2) - This is a very graphic category and you need not know whether the eye is

functionally blind. It generally refers to opaque eyes. The nature of the opacity is not important

here.

"Missing" (Ml, M2) - An eye is actually missing from the fish.

"Other" (OT) - Any manifestations which do not "fit" the above. Describe it in the

remarks column.

Fins;

It must be remembered that this particular assessment system is concerned primarily with health and

condition. It is not concerned with aesthetic values. Eroded or "ragged" fins are definitely indicative

of a departure from normal condition and health. Previously eroded fins which are completely healed

over and showing no evidence of the active erosion are, for the purposes of this assessment, considered

normal. The evaluation of fins is relative to the degree of active erosion process in evidence. For the

purposes of this procedure, the number and location of fins involved is not significant. If only one fin

is displaying active erosion, the observation must be ranked and recorded. If several fins are displaying

erosion with unequal severity, the observation must refer to the most severe in evidence. This unequal

nature of the observations, in this case, is less significant in a full 15 fish sample. The classification is

as follows:

No active erosion (0) - Normal appearing fins with no active erosion. This would include

previously eroded fins which were completely healed over.

Mild active erosion (1) - Active erosion process but no hemorrhage or secondary infection

in evidence.

Severe active erosion (2) - Active erosion with hemorrhage and/or secondary infection

in evidence.

Note! Make a general remark relative to which fins were involved and any other observation of

special significance. There is a space for this type of entry at the bottom of the data collection

worksheet. This is particularly important in the summary.

Opercles;

It is necessary only to observe the degree of shortening of the opercles. The classification is as follows:

Normal opercle (0) - No shortening; gills completely covered

Mild shortening (1) - Slight shortening of the opercle with a very small portion of the gill

exposed
SOP XXXI

Revision No. 2

Date: December 30,1994

Page 4 of 12

Severe shortening (2) - Severe shortening of the opercles with a considerable portion of the gill

exposed.

Gills;

Normal (N) - No apparent aberrations in gills. Be very careful in this observation. The gill

can easily be effected by the manner in which the fish is handed during and after collecting. Be

alert and aware.

"Frayed" (F) - This generally refers to actual erosion of tips of gill lamellae resulting in "ragged"

appearing gills. Mere separation of gill lamellae can be construed to be "frayed" but that

condition may have been caused by something as simple as the manner in which the gill was

exposed by the investigator.

"Clubbed" (C) - This refers to swelling of the tips of the gill lamellae. They can often appear

bulbous or "club" like. The causes are not pertinent until interpretation is considered.

"Marginate" (M) - A graphic description of a gill with a light discolored margin along the distal

ends or tips of the lamellae or filaments. Margination can be and often is associated with

"clubbing". If both seem to apply it is not a problem. It is important that you noted that it was

not normal. Use the one which seems most appropriate.

"Pale" (P) - This refers to gills which are definitely very light in color. Severe anemia_ can

result in gills which are discolored to the point of being white. Severe bleeding induced during

sampling of blood can also result in somewhat pale gills. Gills begin to pale somewhat after

death also. This is not uncommon in fish taken from nets. All of this should be considered in

making the observation.

Other (OT) - Any observation which does not fit above. Describe in remarks.

Pseudobranchs;

Normal (N) - The pseudobranch is located dorsally and anterior to the gills in the branchial

cavity and can be easily observed under the operculum. The normal pseudobranch is quite "flat"

or even concave in aspect and displays no aberrations.

Swollen (S) - The "swollen" pseudobranch is convex in aspect and not difficult to discern upon

close examination.

Lithic (L) - Mineral deposits in pseudobranchs, manifested by appearance of white, somewhat

amorphous spots or foci.

Swollen and Lithic (S&L) - Lithic pseudobranchs are often also swollen.
SOP XXXI

Revision No. 2

Date: December 30,1994

Page 5 of 12

Inflamed (I) - This is a generic use of the term, inflamed, and would more appropriately be

termed "redness" because it also includes observations of hemorrhage and any other cause of

redness. The term, "inflamed" has been traditionally used to describe this condition and is thus

continued for that reason.

Other (OT) - This term will cover any manifestation observed in the pseudobranch which is

not covered in the categories. Be sure to describe in remarks.

Thymus;

Assessment of the thymus involves degree of petechial or "pinpoint" hemorrhage.

No Hemorrhage (0) - The thymus displaying no hemorrhage is considered to be a normal

condition, although this assumption is still under investigation. Caution must be exercised here

because when the thymus involutes or ceases to function there is no observable petechial

hemorrhage. This happens normally as the fish mature. In salmonids, involution of the thymus

is thought to happen at two or three years of age but there is considerable disagreement among

investigators about that point.

Mild hemorrhage (1) - A few red spots or petechial hemorrhages in evidence. This might

be only two or three small spots.

Moderate hemorrhage (2) - More than a few red spots.

Severe Hemorrhage (3) - Many "pin point" hemorrhages in evidence with some of them

coalescing. The general area may also have a swollen tumescent appearance but that should be

recorded in remarks.

Internal Examination

Once the external examination has been completed, position the fish "belly up" and, using sharp/blunt

scissors, make a ventral cut from the anal vent forward to the pectoral girdle. Do not insert the scissors

so far that the internal organs are damaged. In largemouth bass and other centrarchids carefully cut

through the pectoral girdle to lay open the pericardial cavity and expose the heart and aorta. In catfish

leave the pectoral girdle intact.

Liver;

Upon opening the fish, immediately inspect and record the color and condition of the liver. This must

be done prior to collecting blood. Collecting blood and even minor hemmorhaging sustained during

dissection can quickly drain color from the liver.
SOP XXXI

Revision No. 2

Dale: December 30,1994

Page 6 of 12

Normal (A). Good, solid red color. ,

Lighter-or less vivid red color. Not so pale as to be classed general discoloration. Still

considered to be normal.

"Fatty" liver (C). Light tan color, such as "coffee with cream".

Nodules (NO) Nodules in the liver, i.e. white mycobacterial cysts and incipient nodules, such

as those in hepatoma.

Focal discoloration (DF). Change of color in local areas or foci of the liver.

General Discoloration (DG). Color change in the whole liver.

Other (OT). Aberration or deviation in liver which does not fit into above scheme. Class as

OT and describe in remarks.

Blood

Following inspection of the liver, collect blood. Blood is collected with a sharpened, heparinized

microhematocrit tube via a cardiac puncture. In bass blood is tapped from the aorta. By inserting a

finger into the buccal cavity an gently pressing upwards, the heart and aorta can be immobilized against

the roof of the pericardial cavity, making cardiac puncture much easier. In catfish the same technique

can be used to immobilize the heart and aorta, but, since the pericardial cavity is not open, the cardiac

puncture is more difficult. Once the heart and aorta are immobilized between the floor and raised roof

of the pericardial cavity, the sharpened capillary tube is thrust through the membrane separating the

pericardia! and coelomic cavities and into the pericardial cavity in the suspected location of the aorta.

If the aorta is punctured, a spurt of dark red blood will partially fill the tube. If a clear yellow or pale

red fluid enters the tube, the aorta was missed. Try again with a fresh hematocrit tube. When the aorta

has been punctured, blood can be pumped into the tube by gently releasing and then reapplying pressure

on the aorta with the finger still inserted in the buccal cavity. The tube once filled is sealed with

hematocrit clay and placed in a numbered rack. Make every effort to ensure samples are matched with

the appropriate fish. After collecting a blood sample, proceed with the autopsy as outlined below.

Note: After 30 minutes or the collection of 6 to 8 samples, whichever comes first, place tubes in a

hematocrit centrifuge and spin for five minutes. Hematocrits should be read within 2 hours of collection

or before the blood begins to coagulate. To read hematocrits:

Obtain an hematocrit reader and place the filled hematocrit tube so that the bottom of the red

portion is at the 0 line and the meniscus of the clear plasma is at the 100% percent mark. The

packed red cell volume of the blood is the hematocrit and is read as a percentage of the total

blood volume.
SOP XXXI

Revision No. 2

Dale: December 30,1994

Page 7 of 12

The leucocrit is the buffy of gray zone which consist of the leucocytes or white blood cells an

is also read as percent of the total volume on the hematocrit reader.

The protein content of the plasma is determined using a protein refractometer. Take the

hematocrit tube and break it off just above the buffy, gray zone to obtain the clear plasma and

express the plasma onto the clear glass surface of the protein refractometer. Avoid placing any

glass fragments from the tube on the refractometer. Read the weight/volume percent of protein

in the refractometer. The protein refractometer should be calibrated prior to use with distilled

water.

After collecting a blood sample, proceed with the autopsy by examining, in order, the following tissues

and organs, recording the condition of each tissue and organ with a number or letter code listed below:

Mesenteric Fat;

0 - No fat deposited around the pyloric caeca.

1 - Slight, where less than 50% of each cecum is covered with fat.

2 - 50% of each cecum is covered with fat.

3 - More than each cecum is covered with fat.

4 - Pyloric caeca are completely covered by a large amount of fat.

Spleen;

Black (B) - The "black" is actually a very dark red color of the spleen.

(Red coloration of the spleen. There is subjective variation among investigators

as to whether the spleen is black or red, but both condition are considered

normal.)

Granular (G) - Granular or "rough" appearance of the spleen. This is also considered normal.

Nodular (NO) - The spleen contains or manifests fistulas or nodules of varying sizes. These are

often cysts, such as those encountered with mycobacterial infections.

Enlarged (E) - Spleens can, on occasion, be significantly and noticeably enlarged.

Other (OT) - Occasionally there are observable, gross aberrations which do no fit the above.

There may be spleens with a gray mottling and some with very small spleens. These should be

classed as "other" and described in remarks.

Hindgut;
SOP XXXI

Revision No. 2

Dale: December 30,1994

Page 8 of 12

A short distance of the hind gut should be "opened". This should, in fact, have been accomplished as

mentioned above when the body cavity was incised. If not, it must be opened to expose the "inner

lining" or mucosa. Using the handle of a forceps or some other appropriate blunt instrument, lightly

"scrape" out the contents of the hindgut so that you can observe relative reddening or inflammation.

0 - No inflammation or reddening of the hindgut.

1 - Mild or slight inflammation or reddening of the hindgut

2 - Moderate inflammation of the hindgut.

3 - Considerable, severe inflammation or reddening of the hind gut.

Kidney;

Normal (N) - Good firm dark red color lying relatively flat dorsally in the visceral cavity

along the length of the ventral surface of the vertebral column. It will be necessary to pull

the swimbladder and some of the mesentery aside to expose the kidney to view.

Swollen (S) - Enlarged or swollen wholly or in part.

Mottled (M) - Gray discoloration, mottled or "patchy" in appearance ranging from scattered

patches of gray to total gray discoloration. This is not to be mistaken with the superficial

gray appearance induced by the mesenteric membranes on the surface of the kidney. This

should be moved aside before observation is recorded.

Nodular (NO) - The kidney may have a "granular" or nodular appearance and texture. This

may be induced by granulomatous concretions.

Urolithiasis (U) - This condition is also known as nephrocalcinosis and involve mineral

material in the deposition of a white or "cream-colored" amorphous tubules of the kidney, it

can range in appearance from very small white spots to severe involvement with very large

"serpentine" deposits. These sites of deposition are not to be confused with the Stannius

bodies corpora of Stannius which are present in salmonid kidneys and have endocrine

function. The Stannius bodies are generally not associated with the tubules and usually occur

at the "edges" in an area about midway along the kidney. They appear more globular than do

the urolithic deposits.

Other (OT) - This is used to class any aberrations which do not fit into the above scheme.

Record it as OT and describe it in the remarks.

Gall Bladder -

0 - Yellow, empty of partially full

1 - Yellow, full or distended
SOP XXXI

Revision No. 2

Dale: December 30,1994

Page 9 of 12

2 - Grass green

3 - Dark green

Sex;

Male (M) - Testes observed

Female (F) - Ovaries observed Note if "ripe" with eggs

Unknown (U) - immature or undeveloped gonads

General Observations and Remarks

Anything which appears to be abnormal should be noted. It is recommended that the mesenteric

tissues in the visceral cavity be checked for hemorrhage and inflammation and if these conditions are

present they should be so noted in general remarks.
SOP XXXI

Revision No. 2

Date: December 30,1994

Page 10 of 12

D. Scoring

Once autopsies are completed, transcribe the number and letter codes assigned to the condition of

each tissue/organ to a numerical score according to the table below. Note that no score is given to

sex, fat deposits, and gall bladder. Conditions associated with these tissues/organs are not considered

pathological but simply a physiological or nutritional state that varies on a seasonal or other periodic

basis.

Tissue or Organ

Fins

Condition

N

E1.E2

H1.H2

B1.B2

M1.M2

OT

0

1

2

Score

0

30

30

30

30

30

0

10

20

Tissue or Organ

Spleen

Hind Gut

Condition

B

GR

NO

E

OT

0

1

2

3

0

0

30

30

30

0

10

20

30

Opercles

Gill

Pseudobranchs

0

1

2

N

F

C

M

P

OT

N

S

L

SL

I

OT

0

1

2

3

0

10

20

0

30

30

30

30

30

0

30

30

30

30

30

0

10

20

30

Liver

Blood:

Hematocrit

Leucocrit

Plasma Protein

N

S

M

NO

U

OT

A

C

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DF

DG

OT

<4

30c.>6

>6

<3

0

30

30

30

30

30

0

30

30

30

30

30

0

30

0

30

0

30

30
SOP XXXI

Revision No. 2

Date: December 30,1994

Page 11 of 12

E. Calculations and Summary Data

The following calculations are useful in assessing the overall fish health. The specific calculations

performed will be determined by the specific objectives of a given study. Many of the following

calculations can be done by hand, but for most studies the use of a computer or at least pocket

calculator with a function key for calculating mean and standard deviation is recommended.

Ktl;

The value for Ktl is a ratio of length to weight. It is routinely expressed as Ktl x IO5 to

mitigate the problem of carrying so many decimal places in the records. Use the following

equation to obtain a value:

W x 10s

Ktl = L3

Where:

W = Weight in grams

L = Total length in millimeters

Mean and Standard Deviation;

The mean is determined by totaling all of the values for the observations and dividing by the

number of the observations. In-depth discussion of the standard deviation is beyond the scope

of this presentation. Again, a pocket calculator or computer is recommended for making this

calculation. In each sample of fish examined calculate means and standard deviations for the

following observations:

Length

Weight

Ktl

Hematocrit

Leucocrit

Plasma protein

These calculations are made on raw data from autopsy sheets, not from assigned scores.

Fish Health Index (FHT) Score;

Calculate an FHI score for each fish by totaling the numerical score for each tissue/organ.

For a given sample or population of fish calculate the mean and standard deviation from the

individual FHI scores for fish in that sample. The mean FHI scores between samples can be

compared statistically.
SOP XXXI

Revision No. 2

Dale: December 30,1994

Page 12 of 12

Values as Percents of Total Sample

This calculation expresses as a percent the number of fish in a sample with a specific

physical/pathological condition. As an example, consider the eyes. The number of fish with

normal eyes in a given sample is divided by the total number of fish in the sample (x 100) to

yield the percentage of normal-eyed fish in that sample. Similarly, the percentage of fish in a

sample with one blind eye (Bl) can be calculated. A percentage can be calculated for any or

all physical/pathological conditions. It is particularly useful for pinpointing a high incidence

of disease in any one particular tissue or organ.

F. Interpretation

The state-of-the-art in this area has not been refined. Normal and abnormal conditions for each tissue

and organ have been fairly well defined, but measures of overall fish health based on FHI scores, for

example, are still being evaluated. The interpretation of overall fish health is complicated by the

varying sensitivity of different fish species to environmental stresses. A viable strategy for

investigating potential hazardous waste sites is to statistically compare the mean FHI score for a fish

sample from a suspected "impacted" site with the mean score for fish from an established or selected

"unimpacted" reference or background site on a species to species basis. Other strategies may prove

equally enlightening as more FHI studies are completed.
APPENDIX E

AUTOPSY FIELD DATA SHEETS
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Weight:

EPA Region IV

AUTOPSY CLASSIFICATION KEY

Total length in millimeters

Weight in grams

Ktl:

Sex;

Eyes;

Opercles;

Gills:

Pseudobranch:

Thvmus;

Fat:

F

M

W x 10s

L

3

F, note if ripe

EXTERNAL EXAMINATION

Normal (N)

Exopthalmic (popeye) (E1,E2)

Hemorrhagic (H1,H2)

Blind (B1,B2)

Missing (M1,M2)

Other (OT)

No active erosion; previous erosion healed over (0)

Mild erosion with no bleeding (1)

Severe active erosion with hemorrhage and/or

secondary infection (2)

No shortening (0)

Mild shortening (1)

Severe shortening (2)

Normal (N)

Frayed (F)

Marginate (M)

Clubbed (C)

Pale (P) Other(OT)

Normal (N)

Swollen (S) Lithic (L)

Swollen and Lithic (SL)

Inflammed (I) Other (OT)

No hemorrhage (0)

Mild hemorrhage (1)

Moderate hemorrhage (2)

Severe hemorrhage (3)

INTERNAL EXAMINATION

None

Slight, < 50% of each cecum is covered

50% of each cecum is covered

>50% of each cecum is covered

100% of each cecum is covered

(0)

(D

(2)

(3)

(4)

Spleen; Black, red (B)
Hindqut;

Kidneys:

Liver;

Gallbaldder:

Blood:

Hematocrit

Leucocrit

Granular (G)

Nodular (NO)

Enlarged (E)

Other (OT)

No inflammation (0)

Slight inflammation (1)

Moderate inflammation (2)

Severe inflammation (3)

Normal (N)

Swollen (S)

Nodular (NO)

Other (OT)

Mottled (M)

Urolithic (U)

Red, pale red (A)

Coffee & Cream (C)

Nodular (NO)

Discoloration, focal (DF)

Discoloration, general (DG)

Other (OT)

Yellow, empty or partially full

Yellow, full or distended

Grass green

Dark green

(0)

(1)

(2)

(3)

Volume of red blood cells (erythrocytes) expressed

as percent of total blood volume. Centrifuged 5

minutes.

> 31 is normal < 31 is abnormal

Volume of white blood cells (leucocytes) expressed

as percent of total blood volume. "Buffy" zone of

the packed cell column.

< 4 is normal > 4 is abnormal

Plasma protein Amount of protein plasma, expressed as gram percent

(grams per 100 ml).

> 3 to < 6 is the normal range

< 3 is abnormal

> 6 is abnormal
APPENDIX F

SPREADSHEETS SUMMARIZING OF AUTOPSY DATA

AND CALCULATED FHI SCORES
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APPEISTDIX G

Human Health Risk Evaluation of

Fish Data. Mobile River Study
-to sr,.,

*o

} UNITED STATES ENVIRONMENTAL PROTECTION AGENCY

REGION 4

315 COURTLAND STREET N E

ATLANTA GEORGIA 30365

January 30, 1995 -' •

4WD-OHA

MEMORANDUM

SUBJECT: Human Health Risk Evaluation of Fish Data. Mobile River

Study

FROM: Kevin P. Koporec, Toxicologist/^

Office of Health Assessment

THRU: Elmer W. Akin, Chief

Office of Health Assessment

TO: Joanne Benante, RPM

South Superfund Remedial Branch

Per your request, I have evaluated the health risks

associated with potential human consumption of fish from the

Mobile River.

The data forwarded to me for evaluation report levels of

mercury and DDT/DDE/DDD in fish obtained by' EPA for the Mobile

River Study project. From observing the fillet fish data,

mercury and DDT/E/D are reported at higher concentrations in the

largemouth bass than in the catfish fillet samples. In both

catfish and largemouth bass, the fish classified in the "large"

category had the highest concentrations of each of the reported

chemicals.

Rather than attempt to estimate the risks from fish'

ingestion for each sampling station, I have used exposure

assumptions and toxicity values to backcalculate risk-based

concentrations which can then be compared to the chemical

concentrations found at the various sampling stations.

Exposure Assessment- Fish Ingestion

From personal communication with officials at the Alabama

Department of Public Health it was determined that, as an

estimate of reasonable maximum exposure, an individual

subsistence fisherman could consume up to two fish meals per week

from this area of the Mobile/Tombigbee Rivers. While many

individuals may not consume this quantity of Mobile/Tombigbee

River fish, the purpose of this assessment is to evaluate the

potential health- risk in a conservative (health-protective)

manner. This ingestion rate for an adult of 2 fish meals (145 g
per meal) per week from the contaminated source is assumed to

occur 52 weeks per year for a total of 30 years. The exposure

assumptions, other than the ingestion rate, are EPA baseline risk

assessment default values for fish ingestion. All exposure

assumptions are listed in Table 1.

Table 1. Exposure Parameters

Exposure Parameter

Fish Ingestion Rate (IR)

Exposure Frequency (EF)

Exposure Duration (ED)

Body Weight (BW)

Averaging Time (AT)

!i

Assumed Value

0.145 kg /meal

104 meals/year

30 years

70 kg

ED x 365 d/y (noncarcinogens)

70 y x 365 d/y (carcinogens)

Toxicity Assessment- Chemicals detected in Fish

EPA has classified DDT, DDE, and DDD as B2 (probable human)

carcinogens, and has verified cancer slope factors for each of

these compounds. Mercury is class D (not classifiable as to

human carcinogenicity). From the available noncarcinogenic

toxicity data EPA has established oral reference doses (RfDs) for

mercury (both inorganic and methyl) as well as for DDT. The

available toxicity values are listed in Table 2.

Table 2. Toxicity Values

I

Chemical

Mercury'

Reference Dose (mg/kg-d)

[Source]

3E-4 [HEAST/IRIS]'

Slope Factor (kg-d/mg)

[Source]

NA

I

j 3 4E-1 [IRIS]

'Mercury is class D for carcinogenicity; therefore, the slope factor is not

applicable the reference dose used is the same for inorganic [HEAST] and

methyl [IRIS] mercury

" The Guidance For Assessing Chemical Contaminant Data For Use In Fish

Advisories (EPA 1994 ^ recommends adding tne concentrations of DDT, DDE, and

DDD ana comparing to tne nealth based values for DDT.

Derivation of Risk-Based Concentrations for Fish Ingestion

The risk-based concentration is determined by appropriately

combining parameters from the exposure as^v-ssment and toxicity

assessment along with selected target risk values
The following equations result in chemical concentrations in fish

for the carcinogenic and noncarcinogenic endpoints, respectively.

= TCR x BW x AT

IR x EF x ED x SF

= THQ x BW x AT x RfD

IR x EF x ED

where: CF,., = carcinogenic risk-based concentration in fish

CF^ = noncarcinogenic risk-based concentration in fish

TCR = target carcinogenic risk

THQ = target hazard quotient (noncarcinogenic endpoint)

SF = carcinogenic slope factor

> RfD = reference dose

All exposure parameters are defined in Table 1.

The resultant risk based values are listed in Table 3.

Table 3. Risk-based fish concentrations for chemicals detected in

Mobile River Study (mg/kg)

Target

Risk Level

TCR = 1E-6

TCR = IE -5

TCR = IE -4

THQ =0. 1

j THQ =1.0

THQ = 10.0

Chemical

DDT

0.01

0.1

1.0

0 .08

0.8

8

Mercury

NA

NA

NA

0 .05

0.5

5

TCR = target carcinogenic risk (Upperbound)

THQ = target hazard quotient (noncarcinogenic endpoint)

NA- carcinogenic endpoint not applicable for mercury

Risk Characterization- Fish Ingestion

Note: The Upperbound risk values given in the following text

assume a consumption rate as stated in Table 1 of fish which are

all contaminated to the level stated.

Of the reported mercury concentrations for large channel

catfish fillets, station 2 had the highest concentration. The
average of the reported mercury concentrations at station 2 (0.28

ppm) corresponds to an hazard quotient (HQ) between 0.1 and 1.0,

based on the exposure assumptions used in this assessment. The

maximum fillet mercury concentration reported in the study (0.54

ppm) is approximately equal to an HQ of 1.0. For the samples of

largemouth bass- samples classified as large, station 2 also had

the highest reported concentrations of mercury. Both the average

and maximum levels at this station (0.36, 0.41 ppm, respectively)

correspond to an HQ between 0.1 and 1.0.

The sum of the DDE/DDD/DDT concentrations are highest at

station 7 for both the large channel catfish and the large

largemouth bass. In large catfish, the total of the DDE/DDD/DDT

averages at station 7, 0.302 ppm, corresponds to an upper-bound

carcinogenic risk between IE-5 and IE-4, and to a HQ between 0.1

and 1.0. The total of the maximum DDE/DDD/DDT concentrations in

catfish at station 7, 0.409 does not differ noticeably from the

average concentration in terms of estimated risk. For ingestion

of largemouth bass, the total of the DDE/DDD/DDT averages at

station 7, 0.517 ppm, also corresponds to an upper-bound

carcinogenic risk between IE-5 and IE-4, and to a HQ between 0.1

and 1.0. The total of the maximum DDE/DDD/DDT concentrations in

large fillet bass at station 7, 1.35 ppm corresponds to an upperbound carcinogenic risk of approximately IE-4, and to a HQ

between 1.0 and 10.0.

If an adolescent were to consume this fish at a comparable

rate to the adult, the total dose for the no/ncarcinogenic

endpoint would be higher (up to 2-fold) due to lower assumed body

weight for the adolescent. For the carcinogenic endpoint

(DDT/DDE/DDD), however, this difference is negated by the lower

exposure duration for the adolescent (9 yrs) being averaged out

over the lifetime (70 yrs).

Surface Water/Sediment

The surface water and sediment chemical data show detections

of mercury and a few pesticides. The concentrations reported

indicate no significant additional health risk under any

foreseeable exposure scenarios.

Conclusions/Considerations

In considering risk management decisions for this site, I

would emphasize the conservative (health protective) approach

used in deriving the health-based concentrations for DDT and

mercury. The assumption is made in this assessment that a 70 kg

adult regularly eats fish from the same contaminated source,

twice per week, 52 weeks per year, for 30 years and that all fish

eaten have the given level of contamination. Adjustment of

exposure values based on better additional site-specific

information would alter the risks estimates accordingly, most

likely downward.
FDA has set action levels for edible fish for both mercury

(1.0 ppm) and DDT (5.0 ppm). The maximum concentrations of both

mercury and DDT (including metabolites) reported for the Mobile

River Study are below these FDA levels. It should be noted,

however, that FDA's action levels assume that the individual will

consume no more than 1.2 Ib per year of contaminated fish (10% of

the total fish diet from the marketplace). FDA action levels are

not meant to protect the local recreational or subsistence

fisherman. In contrast, the subsistence-type fisherman assessed

in this report is assumed to consume more than 33 Ib of

contaminated fish per year.

If further questions arise, I can be reached at 347-3555,

vmx 6364.

K. P. Koporec/kpk:4WD-OHA: 1586/01/30/9 5/C: \WP51\iMRSFSH.RAl
APPENDIX H

RESPONSIVENESS SUMMARY
MOBILE RIVER STUDY

RESPONSIVENESS STUDY

FOR MAJOR COMMENTS

Human Health Risk Evaluation

Comment 1: The report says that a subsistence fisherman is at risk from

eating large largemouth bass from Station MT-7. this conclusion

is based on some very conservative and highly unlikely

assumptions. It is highly probable that a person consuming 0.145

kg of fish at each of 2 meals per week for 30 years would fish at

only a single location (such as MT-2 or MT-7), or would catch and

eat only the largest largemouth bass. Fish tissue concentrations

should be evaluated, and potential risk estimated, on the basis of

area, or multiple-station, means. Since the presence of a boat

launch near MT-7 presumes that subsistence fisherman are in the

area, and would not be likely to fish the entire 53-mile study

area, we (ADEM) calculated Hazard Quotients and Cancer Risk based

on the average of all fillets from MT-7, MT-8, and jMT-9. Using

these data, the HQ would be 0.108, and the cancer risk would be

1.7 x IO-5 .

Response 1: EPA's risk assessment guidance establishes protocol for an

individual, as having an average cancer risk of 1.0 x IO"6 or a 1

in a million chance of developing cancer. In the MRS Report a

subsistence fisherman scenario was evaluated which yielded a

cancer risks between 1.0 x 10'* and 1.0 x 10"5. Both the MRS

evaluation and the evaluation noted in the comment, show that a

subsistence fisherman as an increased risk of developing cancer,

(e.g. a range between 1 in 100,000 to 1 in 10,000). Therefore,

that ADEMs calculations are consistent with MRS regarding the

increased risk to a subsistence fisherman. Regarding the comment

relating to the probability of a subsistence fisherman eating fish

from only a single location (such as MT-2 or MT-7), EPA is aware

of numerous fish camps along the 53 mile study area. The Walker

Fish Camp, is located immediately down gradient from station MT-7.

A newspaper article in the May 2, 1995 edition of the Mobile

Register (attached) reports that there are individuals and

families that live in trailers at the Walker Fish Camp, Some of

whom have lived their for 20 years. It seem highly feasible that

these individuals may fish and eat fish from that location on a

consistent basis. It is reasonable to assume that if a fisherman

finds a good location to fish he/she will consistently return to

that location to fish. •

Comment 2: EPA has greatly overestimated potential human health risks in the

study area. Even so, the risk estimate is still within the

acceptable risk range of 1 x IO"4 to 1 x IO"6.

Response 2: For clean up actions under the remedial program of CERCLA, the NCP

establishes a risk range of 1 x IO"1 to 1 x IO"6. The NCP states

that EPA's preference is to select remedies that are at the more

protective end of the risk range. Therefore when developing its

preliminary goals, EPA uses 1 x IO"6 as a point of departure.

This means that a cumulative risk level of 1 x IO"6 is used as the

starting point (or initial protectiveness goal) for determining

the most appropriate risk level that alternatives should be

designed to attain. The use of 1 x IO"6 expresses EPA's

preference for remedial actions that result in risks at the more

protective end of the risk range, but this does not reflect a

presumption that the final remedial action should attain such a

.risk level. The 1 x IO"6 point >t departure may be revised to a

different risk level within the acceptable risk range based on the
consideration of appropriate factors including, but not limited

to: exposure factors, uncertainty factors, and technical factors.

For the MRS, the risk assessment calculations yielded a current

risk from carcinogens between 1 x IO"4 to 1 x IO"5. This equates

to some increased risk of developing cancer. The Mobile and

Tombigbee rivers are bodies of water that not only are

recreational, for swimming, boating, and fishing, but are

commercial fisheries. Protectiveness is of the utmost concern.

Based on numerous factors, the MRS Report did not conclude that a

remedial action was necessary for all or part of the study area,

but merely recommended additional monitoring of the river to

verify that contaminant levels continue to decrease to more

acceptable levels. The study also emphasized that the four

Superfund site must continue to address their contaminated

wetlands and full fledged ecological and human health risk

assessment be completed on those sites. EPA does not believe the

potential human health risk has been greatly overestimated, but

was a well balanced study based on sound scientific practices and

protocols.

Comment 3: EPA only used large fish fillets in their calculation of human

health risk for mercury. Use of medium-sized fish fillets in

calculating the concentration term yields a exposure point

concentration below the maximum value for catfish fillets so that

in fact the health risks are even lower than estimated by EPA.

EPA did not state whether inorganic, organic, or total mercury was

used in estimates for human health risk.

Response 3: As part of the MRS, EPA determined a size range for fish

specimens. For largemouth bass, 170 to 225 mm was considered a

small size, 226 to 300 mm was medium, and 301 to 400 mm was large.

For channel catfish, 200 to 275 mm was considered a small size,

276 to 375 mm was medium, and 376 to 500 mm was large. The MRS

human health risk assessment followed the guideline of the Risk

Assessment Guidance for Superfund; the Highest contaminant

concentrations were used in the calculation, because it is

reasonable that an individual could continue to eat fish from the

same location, and because the data set was not excessively large.

This concentration was from a fish composite sample and not an

individual fish and therefore represented an average concentration

of the contaminant for five fish. This is in accordance with the

Agency's directive toward protectiveness. It is important to note

here that EPA collected fish greater in size than was determined

as "large" for MRS. These extra-large fish, not used in the

concentration term, had higher concentration of DDT and mercury

than the large fish specimens. It would seem likely that a

fisherman would keep and eat the largest fish captured and throw

back the smaller specimens.
Preliminary Risk Evaluation

Comment 4a: For ecological risk, the conclusions in the MRS that a potential

for ecological risk is clearly demonstrated is questionable at

best. Assumptions for the ecological effects appear excessively

conservative, and the additive effect of overly conservative

assumptions paints an unrealistic picture of the risk to

piscivorous birds.

Comment 4bs EPA's Summary and Conclusions overstates the ecological risks that

are indicated by the data, unnecessarily raising public concerns

regarding ecological risks. For example, the assumption that a

kingfisher ingests food at a rate of 1.75 times its body weight is

excessively conservative.

Comment 4c: Exposure assumptions for ecological risk assessment are

inconsistent with information published in the scientific

literature and with documentation issued by EPA to support risk

characterization. For the MRS EPA has chosen an unjustifiably

high rate, which significantly inflates risk estimates.

Response 4: EPA did not conduct a comprehensive ecological risk assessment as

part of the MRS. Rather, we produced a limited preliminary

ecological risk evaluation with the purpose of determining if data

produced by this study showed any indication of whether a more

information to determine detailed ecological risk assessment may

be warranted. Moreover, EPA believes any risk assumptions used in

the MRS Report are reasonable, protective of human health and the

environment, and consistent with preliminary ecological risk

evaluations EPA has developed for Sites in Region IV and

nationwide. We do not believe our assessment is "excessively or

overly conservative," "unrealistic," or "inconsistent with

published information." For example, when evaluating the risk to

a piscivious avian, the MRS preliminary risk assessment assumed a

feeding range throughout the 53 mile study area. In fact, the

feeding range for the wood stork is 20 miles and the range for the

kingfisher is smaller at 1.4 miles. If an ecological risk

assessment was undertaken using the smaller feeding ranges and

focusing around the contaminated areas (e.g. the floodplain

wetland of the four Superfund sites), the calculated risk might be

much higher than reported in the MRS.

The documented ingestion rate for a kingfisher is 1.00 - 1.75

times body weight. EPA's use of the 1.75 factor in the risk

calculations is consistent with the Agency's mission of

protectiveness and necessary to eliminate any potential for false

negative results. It is within the range documented iri the

literature and therefore is not "excessively conservative," nor

does it "significantly inflate risk estimates." In fact, a

recalculation using the 1.00 factor instead of the 1.75 factor

still shows a potential risk to the kingfisher and an HQ> 1. EPA

believes the MRS is a well balanced study based on sound

scientific practices and protocols.

Comment 5: The POS states that "for ecological risk assessment a weight of

evidence approach will be followed." EPA's ecological risk

assessment is based totally on calculation of a hazard quotient.

Response 5: The preliminary ecological risk evaluation -relied not only on HQ,

but also on ER-L and ER-M. Moreover, the preamble to the NCP

mentions the "weight of scientific evidence" as a factor relating

to setting preliminary remediation goals. The weight of

scientific evidence concerning exposure is one factor among many
that EPA weighs in making a decision to deviate from the 1 x 10-6

risk level for remediation. The remedial decision or

recommendations made in the MRS relied on the "weight of

scientific evidence" as is envision in the NCP. For the Tombigbee

and Mobile river channel, this remedial decision is for no further

action with continued monitoring to determine that contaminants

continue to decrease to more acceptable levels. For the backwater

wetlands associated with the four NPL sites, the remedial decision

is to address or continue to address these areas as operable units

of the NPL Sites. Since these wetland are a potential contaminant

source to the river, appropriate remedial action will be addressed

through the CERCLA process. It is evident that these decisions

relied on factors including the weight of scientific evidence and

did not strictly rely on the hazard quotients as calculated in the

MRS ecological risk assessment.

Comment 6s Fish samples collected at each of the stations were composited at

three size ranges; no discrete specimens were analyzed. It would

have been useful to analyze some of the larger specimens of large

mouth bass and channel catfish separately. It is suspected that

the concentrations of mercury and total DDTs may be higher in

larger specimens, perhaps to a point where larger adults may

experience reproductive impacts or other health problems.

Response 6: As mention in response 3, EPA did collect fish larger than what

was designated as "large" in the MRS POS. Subsequent to drafting

the MRS report, this "extra large" fish tissue was analyses for.

mercury and DDT. The results showed higher concentrations of

contaminants in the tissue samples of some of these "extra large"

fish compared to the "large" fish designated in the MRS Report

(Figures attached). EPA believes that this analysis supports our

conclusions regarding the human health risk assessment, especially

for a subsistence fisherman, and our ecological risk assessment.
NOAELs

Comment 7a: The rationale for establishing a NOAEL for mercury made reference

to studies done on goshawks, red-tailed hawks, and loons, all with

varying results. The loon data suggests that when the mercury

concentration in the prey was 0.3 to 0.4" mg/kg, a reduction in egg

laying and nesting site and territorial fidelity occurred. The

lower end of this range, 0.3 mg/kg, is equated to a NOAEL of 0.1

mg/kg body weight/day. No rationale is provided for an order of

magnitude change to a NOAEL'of 0.01 mg/kg/day for use with the

wood stork.

Also the rationale for establishing a NOAEL for DDT and

metabolites used data associated with studies on kestrel. ADEMs

calculations equate to a NOAEL between 0.054 and 0.55 mg/kg BW/day

rather than the 0.054 value used in the MRS. It should be noted

that the kestrel, unlike the belted kingfisher, is not a

piscivorous bird.

Comment 7b: EPA's effect level for mercury of 0.01 mg/kg/day is a "no observed

adverse effect level' (NOAEL), it is not a toxicity threshold, but

rather an absolute safe level and therefore is inappropriate.

Comment 7c: The DDE NOAEL selected by EPA underestimates the true avian NOAEL.

EPA selected an avian NOAEL of 0.3 ppm based upon a study in

kestrels. A LOAEL of 3 ppm was also reported in that study. The

true NOAEL is probably between 0.3 and 3 ppm. Therefore, the

NOAEL used in the MRS overestimates the risk.

Comment 7ds The EPA's ecological risk assessment conclusions are not

consistent with available toxicity data. in general, a Hazard

Quotient (HQ) that compares the dose received by the organism in

the environment to a known toxic effect level indicates a risk

threshold. In marked contrast, in the MRS, an assumed NOAEL of

0.01 mg mercury/kg BW/day was used to estimate HQ. As

demonstrated in Section 3, there is a l^rge "safety factor"

inherent in this method, possibly an order of magnitude, when

considered against the toxicity LOAEL. thus, the HQ in the MRS

are not risk thresholds. They are safe exposure levels.

Response 7: A LOAEL is defined as the lowest observed adverse effect level. A

NOAEL is defined as a no observed adverse effect level.

Therefore, by definition, a LOAEL is an effect level. In other

works, an adverse effect is existent at a LOAEL. On the other

hand, a NOAEL is a non-effect level; or no adverse effect is

evident at a NOAEL. Since the mission of EPA is protection human

health and the environment, the only acceptable level would be one

with no adverse effect. Use of a LOAEL would not be protective of

the environment. As far as a "risk threshold", neither a NOAEL

nor a LOAEL are risk thresholds. Since a LOAEL is an effect level

and a NOAEL is a non-effect level, the risk threshold lies

somewhere between the NOAEL and the LOAEL. Therefore used of a

LOAEL to determine protectiveness is inappropriate and EPA's use

of a NOAEL is appropriate and adequately estimated the potential

risk to the species evaluated.

Regarding the loon data presented in comment 7a, a mercury

concentration in prey ranging from 0.3 to 0.4 mg/kg yields a LOAEL

of 0.1 mg/kg/day because these concentrations equate to an

observed effect in the loon. For calculation of a NOAEL, EPA use

a dietary level of 0.03 mg mercury per kg in prey. This

concentration yielded a NOAEL of 0.01 mg/kg/day. The order of

magnitude difference between a LOAEL and a NOAEL is a common
scientific default value based upon years of historical data and

consistent with human health evaluations of acute versus chronic

exposure. Regarding the 0.054 value used in the MRS, the Lincer

1995 study revealed that egg shell thickness was significantly

different at 0.054 mg/kg BW/day from archived egg shell thickness.

Therefore, 0.054 is not a NOAEL because an observed effect was

noted, however, it was the lowest available data point that could

be used to estimate protectiveness. Use of any value above 0.054

would not be protective.

EPA's use of 0.01 mg/kg/day for mercury was based upon a study

where this value yielded an endpoint resulting in a reproductive

reduction in egg laying, nesting, and terrestrial fidelity. The

NOAEL was not based on toxicity.

In the preliminary ecological risk evaluation, EPA, in the absence

of having information on the precise toxicity threshold, has

chosen to NOAELs in calculating HQs. We feel this is appropriate

and the most protective approach.
Use of Non-Detects

Comment 8a: Eliminating from analysis those samples below the limit of

detection inappropriately increases mean values. Including less

than detection samples as either zero or one-half the minimum

quantification limit results in a. mean mercury concentration for

station MT-0 to MT-8 of 0.02 mg/kg rather than the 0.03 mg/kg used

in Tables 33 and 35.

Comment 8b: The decision to exclude from calculation of mean tissue

concentrations those samples which were at or below the MDL is

contrary to EPA policy in most other areas of statistical

evaluation. It is not clear what the authors mean when they say

that the data represents "a conservative treatment of results."

It would be more appropriate if it was an "accurate* treatment of

the data following the Guidance documents cited in the POS.

Comment 8c: At a minimum, EPA should have included all of the study area fish

in order to address the mobility of both fish and fisherman over a

30 year exposure period.

In calculating the means for the data set, the values reported as

a minimum detection levels (MDLs) were not included in the mean.

These means should be calculated with the (MDLs) since this is

still valid data.

Comment 8d: Statistical analysis conducted by EPA did not include non-detect

values. Ignoring values at or below the detection limit when

statistically comparing reference and site locations artificially

increases the chances of finding a difference. Because the

statistical tests used rely on the variance in each of the data

sets being compared, ignoring non-detects artificially reduces the

variance in one or both data distributions such that distributions

that may have overlapped and been statistically indistinguishable

are collapsed such that the distribution no longer overlap and

spurious conclusions are drawn form these mis-applied statistical

tests.

Response 8: EPA has reevaluated the MRS Report and the decision regarding the

elimination of non-detects from the calculation of mean tissue

concentrations. In response to the comments submitted from review

of the draft report, a re-analysis of the data was performed to

include the non-detects in the calculations. The final report was

revised to include the re-analysis and inclusion of non-detects in

the fish mean tissue concentration. Revised conclusions on the

re-calculations are included in the final report.
Statistical Treatment of Fish Tissue Data

Comment 9- The use of Duncan's Multiple Range test was not rationalized in

the text of the report. Since the background sites represent

sections of the river rather than experimental controlled variate,

a model I ANOVA technique, would be more appropriate. A more

concise rationalization for the use of Duncan's test in necessary.

An analysis of the bass DDTr data reveal that the data set does

not satisfy the assumptions of normality or homogeneity of

variances required for ANOVA techniques. It is never stated

within the report whether the data were assessed for either

normality or equal variances. The cumulative frequency figures

presented strongly suggest that the data are skewed. A histogram

of the data and its distribution pattern would be more

enlightening.

Response 9: Data used in the statistical determinations were examined for

normal distribution characteristics and homogeneity of variances.

Where necessary, transformation of the data sets was accomplished

and re-examined to assure normality and homogeneity of variances.

Additional analyses included examination of confidence bounds for

skewness and kurtosis. A model I ANOVA was followed by an

unplanned multiple comparison procedure. Duncan's multiple range

test was selected because of its power to discern differences

between means. The report now reflects these procedures.
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DIHB LANDING

NEAR CHRYSLER. AL

COFFEEVILLE

LOCK & DAM

X (RIVER MILE 116.66)

OLIN AND

CIBA-GEIGY

NPL STIES

RIVER MILE 45

STAUFFER

NPL SITES

RIVER MILE 26

MOBILE

BAY

AHEA OF RIVBB SAMPLED

FOR PISH

FIGURE i

MOBILE/TOMBIGBEE RIVERS

SAMPLING STATIONS AND AREAS

MOBILE RIVER STUDY

1993-1994

SCALE IN MILES

o 5 io
T

DIXIE LANDING

NEAR CHRYSLER. AL

COFFEEVILLE

LOCK & DAM

(RIVER MILE 116.66)

OUN AND

CIBA-GEIGY

NPL SRES

STAUFFER

NPL SITES

RIVER MILE 26

MOBILE

BAY

AREA OF RIVER SAMPLED

FOR FISH

FIGURE I

MOBILE/TOMBIGBEE RIVERS

SAMPLING STATIONS AND AREAS

MOBILE RIVER STUDY

1993-1994

SCALE IN MILES

10
T

DIXIE LANDING

NEAR CHRYSLER. AL

COFFEEVILLE

LOCK & DAM

(RIVER MILE 116.66)

OUN AND

CIBA-GEIGY

NPL SITES

STAUFFER

NPL SITES

RIVER MILE 26

MOBILE

BAY

AREA OF RIVER SAMPLED

FOR FISH

FIGURE 1

MOBILE/TOMBIGBEE RIVERS

SAMPLING STATIONS AND AREAS

MOBILE RIVER STUDY

1993-1994

SCALE IN MILES

10
COFFEEVILLE

LOCK & DAM

(RIVER MILE 116.66)

DIXIE LANDING

AT CHRYSLER. AL

RIVER MILE 60

RIVER MILE 26

MOBILE

BAY

TO BE SELECTED IN

SCREENING PHASE:

\ » - Backwater

':',; - Backwater

FIGURE 1.

MOBILE/TOMBIGBEE RIVERS

STUDY REACH AND SAMPLE

STATION LOCATIONS

SCALE IN MILES

To