EPA/XXX/XXX/XXX

June, 1997

AN ASSESSMENT OF SEDIMENTS FROM THE

UPPER MISSISSIPPI RIVER 

Final Report - June, 1997

by

F. James Dwyer, Eric L. Brunson, Timothy J. Canfield

Christopher G. Ingersoll, Nile E. Kemble

U.S. Geological Survey

Biological Resources Division

Environmental and Contaminants Research Center

4200 New Haven Road

Columbia, Missouri 65201

USEPA Project No. DW14935486-01-0

Project Officer:

 Thomas Armitage

U.S. Environmental Protection Agency

Office of Science and Technology

Washington, DC 20460

U.S. Environmental Protection Agency

Office of Science and Technology

Washington, DC 20460
ii

NOTICE

This is the final report of research funded under USEPA Project No. DW14935486-01-0. 

Mention of trade names or commercial products does not constitute endorsement or

recommendation for use. Data from this report can be obtained electronically from:

anonymous ftp - ftp://ftp.msc.nbs.gov/pub/umr/umr.zip

world wide web - http://www.msc.nbs.gov/pubs/umr.html

For problems with access to the above addresses please e-mail the Webmaster, Chris Henke, at

chenke@msc.nbs.gov or call 573-875-5399.
iii

ABSTRACT

The U.S. Geological Survey (USGS) has been monitoring the Upper Mississippi River

(UMR) since 1987 to document the fate and transport of contaminats associated with sediments. 

The UMR is that part of the river upstream of the confluence with the Ohio River at Cairo, IL

and consists of a series of 26 navigational pools created by a lock and dam system extending

from Minneapolis, MN to St. Louis, MO. The navigational pools are shallow, lake-like areas

which trap and store large quantities of fine-grained sediments during normal river flows. 

Concern with the redistribution of the river sediments arose after the flood of 1993. This project

was designed to evaluate the current status of sediments in the UMR by: (1) measuring the

concentrations of contaminants in sediments of the UMR, (2) evaluating the toxicity of sediments

collected from the river, (3) determining the bioaccumulation of contaminants from UMR

sediments using field-collected and laboratory exposed oligochaetes, and (4) determining the

benthic community structure in fine-grain sediments within the river.

To conduct these assessments, sediment samples and benthic organisms were collected

from 24 of the 26 navigational pools in the river and from one pool in the Saint Croix River. 

Two types of sediment samples were collected from the pools. One sediment sample was a

composite of 15 to 20 sediment grabs along one to five transects across the downstream one-third

of each pool (B samples). The other sediment sample was a composite of grabs from one station

on one transect within each pool (C samples). The latter stations were selected based on

historical chemistry data and the potential to collect oligochaetes. Samples were not collected

from the main navigation channels. Chapter 1 of this report describes whole-sediment toxicity

tests which were conducted for 28 days with the amphipod Hyalella azteca. Survival, growth

and sexual maturation were the measurement endpoints. Toxicity tests were conducted with both

the B and C sediment samples. Chapter 2 describes the bioaccumulation of contaminants from

sediments using field-collected oligochaetes and 28-day bioaccumulation studies conducted in

the laboratory with the oligochaete Lumbriculus variegatus. Bioaccumulation tests were

conducted with 13 of the 24 C sediment samples. Chapter 3 assesses the benthic community in

all 24 C samples. Using the Sediment Quality Triad approach, the status of UMR sediments was

assessed by integrating sediment chemistry, laboratory toxicity tests and benthic community

measurements.

In the toxicity tests, Hyalella azteca survival was significantly reduced in only one

sediment sample (13B) relative to both a control and reference sediment. Growth of amphipods

was also reduced in only one sediment sample (26C). Sexual maturation was not significantly

reduced in any treatments. No correlations were observed between survival, growth or sexual

maturation and any of the physical or chemical sediment characteristics. Using sediment

chemistry and the Effect Range Median (ERM), 96% of the samples were classified as non-toxic

(i.e. measured chemical concentrations rarely exceeded ERMs). Classifications using ERMs and

sediment chemistry were consistent with the biological results from the H. azteca toxicity tests.

In the bioaccumulation tests, concentrations of contaminants were relatively low in native

oligochaetes collected from the pools as well as in oligochaetes exposed to the sediments in the

laboratory. Organochlorine pesticides were generally below detection in sediment and tissue

samples. Only aliphatic and polycyclic aromatic hydrocarbons (PAHs) and total polychlorinated
iv

biphenyls were frequently measured above detection limits in oligochaete tissue and sediment

samples. Concentrations for a specific contaminant in laboratory-exposed and field-collected

oligochaetes were similar within a station. About 90% of the paired PAH concentrations in

laboratory-exposed and field-collected oligochaetes were within a factor of three of one another. 

With the detection limits used to analyze samples, contaminants were detected in tissue samples

more often than in sediment samples. Concentrations of PAHs in oligochaetes collected from the

pools or exposed in the laboratory to sediments from the UMR were up to 1000 times less than

tissue concentrations measured in oligochaetes from highly-contaminated sites within the U.S.

that our laboratory has previously studied.

The benthic community was dominated by oligochaetes and chironomids in 14 of the 23

sediment samples from the UMR and the one sediment sample from Saint Croix River. 

Fingernail clams comprised a large portion of the community in 3 of the samples and exceeded

1,000/m in 5 of the samples. Total abundance values of invertebrates ranged from 250/m ( 2 2

station 1C) to 22,389/m (station 19C) and were comparable to previously reported values for the 2

UMR. The frequency of chironomid mouthpart deformities was only 3% which is consistent

with the incidence of mouthpart deformities from uncontaminated sediments. Correlations

between benthic measures, sediment chemistry or other abiotic parameters exhibited few strong

or significant correlations indicating benthic communities are most likely controlled by factors

independent of contaminant concentrations. 

The Sediment Quality Triad (Triad) is a weight-of-evidence approach used to assess the

contamination of sediments by integrating sediment chemistry, laboratory toxicity testing and

benthic community measures. Results from the Triad analysis indicated 88% of the samples

were classified as not impacted based on sediment chemistry, laboratory toxicity and benthic

measures. These results are consistent with the bioaccumulation study in which concentrations

of contaminants in tissue were less than other U.S. sites that our laboratory has previously

studied. In addition, pools in about the lower third of the river had lower sediment contaminant

concentrations, less accumulation of contaminants in tissue, and greater taxa richness.

Sediments are often both a sink for water-borne contaminants and a source of

contaminants to the overlying water. In addition, sediments may accumulate significant

concentrations of contaminants even when water quality criteria are not exceeded. The results

from the present study indicate that the UMR is not severely contaminated relative to other sites

that have been studied in the U.S. Perturbations that may occur could be attributed to

channelization, sedimentation from surface runoff or long term changes in the natural flow

conditions of the river due to lock and dam construction. This study only conducted a partial

assessment of the UMR sediments and included no assessment of river water. Further, this study

was a one-time assessment that was conducted after a major flood event and does not evaluate

temporal or spatial variability of sediment contamination within the pools. Future research on, or

management of, the Upper Mississippi River should evaluate the limitations of this study. 
v

TABLE OF CONTENTS

List of Figures . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . iv

List of Tables . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . vii

List of Appendices . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . viii

Chapter 1: Evaluation of Contamination in Sediments Collected From

Navigational Pools of the Upper Mississippi River Using a

28-day Hyalella azteca Test. Kemble, N.E., 

E.L. Brunson, T.J. Canfield, F.J. Dwyer, and C.G. Ingersoll . . . . . . . . . . . . . . 1.1

Chapter 2: An Evaluation of Bioaccumulation of Contaminants from

Sediments from the Upper Mississippi River Using 

Field-collected Oligochaetes and Laboratory-exposed 

Lumbriculus variegatus. Brunson, E.L., T.J. Canfield,

F.J. Dwyer, C.G. Ingersoll, and N.E. Kemble . . . . . . . . . . . . . . . . . . . . . . . . . . 2.1

Chapter 3: Assessing Sediment Toxicity From Upper Mississippi

River Navigational Pools Using a Benthic Invertebrate

Community Evaluation and the Sediment Quality Triad

Approach. Canfield, T.J., Brunson, E.L., F.J. Dwyer,

C.G. Ingersoll, and N.E. Kemble . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 3.1
vi

LIST OF FIGURES

Figure 1.1 Map of Upper Mississippi River from Minneapolis, MN to St. Louis, MO . . 1.15

Figure 1.2 Concentrations of simultaneously extracted metal (SEM) cadmium in 

sediment samples compared to effect range median (ERM) for cadmium . . . 1.16

Figure 1.3 Concentrations of SEM Pb in sediment samples compared to ERM for Pb . . 1.17

Figure 1.4 Concentrations of chrysene in sediment samples compared to ERM for 

chrysene . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 1.18

Figure 1.5 Concentrations of benzo(a)pyrene in sediment samples compared to ERM

for benzo(a)pyrene . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 1.19

Figure 1.6 Number of ERM exceedances compared to sum ERM quotient ........... 1.20

Figure 1.7 Survival versus sum ERM quotient samples compared to other known

contaminated sites . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 1.21

Figure 2.1 Diagram of in-line flow splitter used to deliver water ................... 2.12

Figure 2.2 Total accumulation of polycyclic aromatic hydrocarbons by

laboratory-exposed Lumbriculus variegatus .......................... 2.13

Figure 2.3 Total accumulation of polycyclic aromatic hydrocarbons by field-collected

oligochaetes . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 2.14

Figure 2.4 Comparison of tissue concentrations in laboratory-exposed Lumbriculus

variegatus versus field-collected oligochaetes . . . . . . . . . . . . . . . . . . . . . . . . 2.15

Figure 2.5 Ratio of tissue concentrations in laboratory-exposed Lumbriculus

variegatus or field-collected oligochaetes for select PAHS . . . . . . . . . . . . . . 2.16

Figure 2.6 Biota-sediment accumulation factors for laboratory-exposed Lumbriculus

variegatus and field-collected oligochaetes for PAHs from the current

study and calculated from PCB homolog data reported in the literature ..... 2.17

Figure 3.1 Quadrant analysis for Hyalella azteca and sum ERM quotient . . . . . . . . . . . . 3.19

Figure 3.2 Percent chironomid deformities for samples from the Upper

Mississippi River . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 3.20
vii

LIST OF TABLES

Table 1.1 Results of sediment tests with Hyalella azteca ........................ 1.22

Table 1.2 Physical and chemical characteristics of sediments . . . . . . . . . . . . . . . . . . . . . 1.24

Table 1.3 Spearman rank correlations for simultaneously extracted metals and

physical sediment characteristics . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 1.26

Table 1.4 Regression data for amphipod survival, length and sexual maturation

with sediment physical and chemical characteristics . . . . . . . . . . . . . . . . . . . . 1.27

Table 1.5 Regression data normalized to total organic carbon for amphipod 

survival, length and sexual maturation . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 1.29

Table 2.1 List of molecular weights and log Kow for contaminants used to 

compare laboratory-exposed Lumbriculus variegatus and

field-collected oligochaetes . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 2.18

Table 2.2 Biota-sediment accumulation factors for the current study and reported

in the literature ................................................. 2.19

Table 3.1 Percent contribution of each taxa to the overall total abundance estimates . . . 3.21

Table 3.2 Spearman rank correlation for select invertebrate metrics with naphthalene,

chrysene, benzo(a)pyrene, Cd, Ni, Pb, Zn, percent water, percent sand,

percent silt, percent clay and total organic carbon ...................... 3.22

Table 3.3 Summary of quadrant analysis for scores of individual and combined benthic

measures with the sum effect range median quotient . . . . . . . . . . . . . . . . . . . 3.23

Table 3.4 Summary of measures used for the Sediment Quality Triad .............. 3.24
viii

LIST OF APPENDICES

Appendix 1.1 Pore-water quality for whole-sediment tests .......................... 1.30

Appendix 1.2 Mean measured overlying water quality for whole-sediment toxicity tests . . . 1.32

Appendix 1.3 List of polycyclic aromatic hydrocarbons (PAHs) and organochlorines

analyzed for in sediment . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 1.34

Appendix 1.4 Amphipod length data from first study .............................. 1.35

Appendix 1.5 Amphipod length data from second study ............................ 1.44

Appendix 1.6 Amphipod maturation and survival data from first study ................ 1.53

Appendix 1.7 Amphipod maturation and survival data from second study .............. 1.54

Appendix 1.8 Concentrations of simultaneously extracted metals (SEM) , acid volatile

sulfides (AVS) and SEM/AVS . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 1.56

Appendix 1.9 Concentrations of organochlorines in sediment samples . . . . . . . . . . . . . . . . . 1.58

Appendix 1.10 Concentrations of PAHs in sediment samples ........................ 1.60

Appendix 2.1 Mean measured overlying water quality for bioaccumulation tests ......... 2.20

Appendix 2.2 Tissue concentrations of organochlorine compounds measured in laboratoryexposed and field-collected oligochaetes . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 2.21

Appendix 2.3 Tissue concentrations of PAHs measured in laboratory-exposed and

field-collected oligochaetes . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 2.22

Appendix 2.4 Total accumulation of PAHs in laboratory-exposed and field-collected

oligochaetes . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 2.23

Appendix 2.5 Ratio of laboratory to field and field to laboratory tissue concentrations . . . . 2.28

Appendix 3.1 Mean abundance data for Oligochaeta taxa ........................... 3.25

Appendix 3.2 Mean abundance data for Chironomidae taxa ......................... 3.26

Appendix 3.3 Mean abundance data for benthic invertebrate taxa excluding oligochaetes

and chironomids . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 3.27
1.1

Chapter 1: Evaluation of Contamination in Sediments Collected from Navigational

 Pools of the Upper Mississippi River Using a 28 Day Hyalella azteca Test

Kemble, N.E., Brunson, E.L., Canfield, T.J., Dwyer, F.J., and Ingersoll, C.G.

Introduction

The Mississippi River is the largest river system in the United States. Because of its location, the

river receives contaminant inputs from a variety of industrialized and agricultural sources. The

Upper Mississippi River (UMR), the stretch of river upstream from the confluence with the Ohio

River at Cairo, IL, contains a series of 26 navigational pools created by a lock and dam system

from St. Louis, MO to Minneapolis, MN (Rada et al 1990; Figure 1.1). These navigational pools

are shallow lake-like areas which trap and store large quantities (1 to 4 cm/yr) of primarily finegrained sediments during normal river flows (McHenry et al 1984; Nielsen et al 1984). 

Dredging activities, commercial navigation, recreational boating and natural resuspension

processes can result in the remobilization of these sediments. Concern about the resuspension

and transport of these sediments and the contaminants associated with them arose after the flood

of 1993 (Moody and Meade 1995; Moody et al 1996).

The United States Geological Survey (USGS) has been monitoring the transport and

degradation of pollutants in the UMR since the fall of 1987 (Moody and Meade 1995). Studies

have monitored concentrations of contaminants in fish (Hora 1984; Wiener et al 1984),

invertebrates (Beauvais et al 1995; Steingraeber and Wiener 1995), sediments (Bailey and Rada

1984; Wiener et al 1984; Rada et al 1990; Frazier et al. 1996; Ingersoll et al 1997) or a

combination of the three (Peddicord et al 1980; Boyer 1984) in select pools in the UMR. 

However, little information was available on contaminant concentrations and toxicity in sediment

samples throughout the entire pool system of the UMR.

Four studies were conducted to assess the nature and extent of sediment contamination in the

navigational pools of the UMR: (1) contaminant concentrations were measured in sediments

before and after the flood of 1993 (Moody et al 1996); (2) whole-sediment toxicity tests were

conducted (this chapter); (3) whole-sediment bioaccumulation tests were conducted (i.e.; Chapter

2); and (4) benthic-community structure were evaluated (i.e.; Chapter 3). Sediment samples were

collected from June 11th to July 5th, 1994 from pool 1 (near Minneapolis, MN) to pool 26 (near

St. Louis, MO) of the UMR system (Figure 1.1). The objective of the study presented in this

chapter was to assess the toxicity of sediments from navigational pools of the UMR system using

28-day toxicity tests with the amphipod Hyalella azteca, measuring for potential effects on

survival, growth or sexual maturation.

Materials and Methods

Sample Collection, Handling, and Storage

Differential Global Positioning System (GPS) using a local reference was used to locate

sampling stations in the upper pools (1-14) and the Saint Croix River. A differential GPS using
1.2

the navigational beacon near St. Louis, MO., as the reference to locate sampling stations in the

lower pools (15-26). A 3.5 composite sediment sample was collected from each of the 26

navigational pools (pool samples designated as "B" samples; Moody et al 1996). These

composite samples of surface (upper 10 cm) sediments were collected using a van Veen grab

from 15 to 20 stations along one to five transects (typically 3 to 5 stations/transect) from the

downstream one-third of each navigation pool (except pool 17) in the UMR and from one site in

the Saint Croix River (SC) just upstream from its confluence with the Mississippi River in

Wisconsin (Figure 1.1; Moody et al 1996). Samples were not collected from the main

navigation channel which was assumed to contain courser sediment that had been deposited for a

short period of time. A 2-L subsample of the 3.5 L samples for toxicity testing and physical and

chemical characterization were removed and placed in a 2-L high density polyethylene (HDPE)

screw topped container. Samples were stored in a cooler at 4 C for 7 to 14 days on the research o

ship Acadiana, then shipped on ice to the Environmental and Contaminants Research Center

(ECRC - formerly the Midwest Science Center) in Columbia, MO. Two 125-mL subsamples

from each B sample were collected at the start of the toxicity tests for physical (grain size and

TOC) and chemical (organic and metal) characterization. 

A second composite sediment sample was also collected from each pool at one station on one

of the transects (station samples designated as "C" samples). The individual stations © samples)

were selected based on historical chemistry data and the potential for the collection of large

numbers of oligochaetes for bioaccumulation evaluations (Chapter 2). Station sediment samples

(C samples) for toxicity and bioaccumulation (Chapter 2) testing were collected with a Ponar

grab (529 cm area). Each C sample was a composite sample collected from the upper 6 to 10 2

cm of the sediment surface within a 5-m radius area. A total of 35 to 80 L of sediment was

collected from each C station. The sediment was then placed into a 120-L HDPE drum and

homogenized on ship with a stainless steel auger on a hand-held power drill. Subsamples of

these C samples were taken for (1) laboratory toxicity and laboratory bioaccumulation testing (10

L), (2) physical characterization (250 mL) and chemical characterization (250 mL for organics

and 250 mL for metals) and (3) benthic invertebrate assessment (2 L). The remaining C sample

was then sieved and native oligochaetes were collected for bioaccumulation analyses (Chapter 2). 

Sediment samples were stored in a cooler on the ship at 4 C for 7 to 14 days, then shipped on ice o

to the ECRC in Columbia, MO. Once at the ECRC, sediment samples were stored in the dark at

4 C until the start of the study. The control sediment (FLOR) used in the toxicity tests was a fine o

silt- and clay-particle size soil collected near St. Louis MO. This control sediment has been used

in previous studies (Kemble et al 1994).

Culturing of Test Organisms

Amphipods were mass cultured at 23 C with a luminance of about 800 lux according to o

procedures outlined in Tomasovic et al (1995) using 80-L glass aquaria containing 50 L of

ECRC well water (hardness 283 mg/L as CaCO , alkalinity 255 as CaCO , pH 7.8). Artificial 3 3

substrates were also placed in the amphipod culture aquaria (six 20-cm diameter

sections/aquarium of "coiled web material"; 3M Corp., Saint Paul, MN). Known-age amphipods

were obtained by isolating mixed-aged adults in a 5-mm mesh (#35 US Standard size sieve) sieve
1.3

in a pan containing about 2 cm of well water. After 24 h, well water was sprinkled through the

sieve, flushing <24-h-old amphipods into the pan below. These <24-h old amphipods were then

placed into flow-through glass chambers for 10 d before the exposure began. Isolated amphipods

were fed maple leaves and ground Tetramin® ad lib until the start of the test.

Toxicity Tests

Sediment Preparation: Sediment samples were re-homogenized in the laboratory using either a

plastic spoon (for the B samples) or a hand-held power drill with a stainless steel auger (for the C

samples). Subsamples were then collected for: (1) pore-water preparation, (2) physical and

chemical characterizations, (3) toxicity testing, and (4) bioaccumulation testing © samples only;

i.e., Chapter 2).

Water Quality: About 170 mL of pore water was isolated from each sample by centrifugation at

4 C for 15 min at 5200 rpm (7000 x G). A 50-mL subsample for total sulfide determination was o

removed from each sample and preserved with 0.1 mL of 2N zinc acetate solution (APHA 1985). 

Total dissolved sulfide was determined with an Orion EA940 Expandable ionAnalyzer, Orion

94-16 silver/sulfide electrode, and a Orion 90-02 double junction reference electrode. Dissolved

oxygen (mg/L, with a YSI Model 54A oxygen meter and a YSI 5739 probe), temperature ( C) o

and conductivity (μs/cm @ 25 C with a Orion 140 S-C-T meter and a 014010 conductivity cell) o

were determined on the remaining volume. Subsamples of pore water were then removed for the

following determinations: total ammonia (mg/L) with an Orion EA940, and Orion 95-12

ammonia electrode, alkalinity (mg/L, as CaCO ) and pH with an Orion EA940 Expandable 3

ionAnalyzer, Orion 917001 ATC probe, and Orion 8165BN combination pH probe, and total

hardness as (mg/L, as CaCO ) by EDTA titration. Unionized ammonia concentrations (mg/L, as 3

NH ) were calculated by adjusting total ammonia concentrations to pH and temperature using the 3

formula presented in Thurston et al (1979). Hydrogen sulfide concentrations (mg/L) were

calculated by adjusting the total dissolved sulfide concentrations to pH and temperature using the

relationship presented in Broderius and Smith (1977).

Mean characteristics of porewater water quality (ranges in parentheses) are as follows: pH

7.45 (6.69 to 8.17); alkalinity 505 (244 to 852) mg/L; hardness 504 (148 to 852) mg/L; dissolved

oxygen 5.04 (1.50 to 9.35) mg/L; conductivity 906 (380 to 1680) μs/cm @ 25 C; total ammonia 0

5.320 (1.210 to 22.700) mg/L; unionized ammonia 0.007 (0.000 to 0.025) mg/L; total sulfide

0.055 (0.000 to 0.569) mg/L; and hydrogen sulfide 0.023 (0.000 to 0.569) mg/L (Appendix 1.1).

 The following parameters were measured in overlying test water on Day -1 (the day before

amphipods were placed into the beakers) and at the end of each toxicity test: dissolved oxygen,

temperature, conductivity, pH, alkalinity, total hardness, and total ammonia. Methods used to

characterize overlying water quality in the whole-sediment tests were similar to the methods

described for characterization of pore water. Dissolved oxygen, pH, and conductivity were also

measured weekly. Temperature in the water baths holding the exposure beakers was measured

daily. Overlying water pH, alkalinity, total hardness, conductivity and total ammonia

measurements were similar among all stations, the control, and the in flowing test water

(Appendix 1.2). Dissolved oxygen measurements were at or above acceptable levels (>40% of
1.4

saturation; ASTM 1995) in all treatments throughout the study (Appendix 1.2). Means (ranges in

parentheses) of overlying water quality of each parameter are as follows: pH 8.07 (7.58 to 8.72);

alkalinity 87 (59 to 151) mg/L; hardness 128 (111 to 160) mg/L; dissolved oxygen 6.70 (5.84 to

7.53) mg/L; conductivity 392 (359 to 428) μs/cm @25 C; total ammonia 0.416 (0.090 to 1.520) 0

mg/L; and unionized ammonia 0.003 (0.000 to 0.012) mg/L (Appendix 1.2).

Toxicity Tests: All sediment tests were started within three months of sample collection from the

field. Due to the number of samples collected, half of the samples (i.e., half of the sites) were

randomly selected for the initial testing. The second set of sediment samples was tested after

completion of testing of the first set of samples. Sediment samples for the toxicity tests were

homogenized the day before animals were added to exposure beakers (Day -1), using procedures

previously described.

Toxicity tests were conducted with Hyalella azteca for 28 days. Effects of exposure to

sediments on survival, length, and sexual maturation of amphipods were measured (USEPA

1994; ASTM 1995). Each 300-mL beaker contained 100 mL of sediment and 150 mL of

overlying water. The photoperiod was 16:8 h (light:dark) at a light intensity of about 500 lux. 

Four replicate beakers/sample were placed in a ventilated water bath maintained at 23 C. Each o

beaker received 1.0 volume additions/d of overlying water starting on Day -1 (Zumwalt et al

1994). The overlying water used in the sediment toxicity exposures was a reconstituted

moderately hard water (hardness 95 mg/L as CaCO , alkalinity 65-70 mg/L as CaCO , pH 8.0- 3 3

8.3; USEPA 1994). One diluter cycle delivered 50 mL of water to each beaker (diluters cycled

every 8 h ± 15 min). Amphipods were acclimated to the test water over 6 h before exposures

began by sequentially transferring animals at 2 h intervals into 50:50 and 25:75 mixtures of well

water:test water, and then into 100% test water. Tests were started on Day 0 by placing 10

amphipods (10- to 11-d old) into each beaker. The water surface in each beaker was checked 15

min after organisms were placed in the beaker for floating organisms. Amphipods in each beaker

were fed 3 mg of Purina Rabbit Pellets in a water suspension three times a week for the first 7 R

days of the exposure, and 6 mg three times a week for the last 21 days of the exposure. If

excessive mold ($60% sediment surface) was observed on the sediment surface of any of the

beakers in a treatment, feeding was withheld from all of the beakers for that treatment (the

number of feedings withheld ranged from 0 to 5 depending on the treatment; USEPA 1994;

ASTM 1996). Beakers were observed daily for the presence of animals, signs of animal activity

(i.e., burrowing), and to monitor test conditions (i.e.; water clarity).

Amphipods were retrieved from each beaker at the end of exposures using procedures

described in Kemble et al (1994). Surviving organism were combined into a scintillation vial

and preserved in 8% sugar formalin for later measurement of length, and sexual maturation. A

Zeiss® Interactive Digital Analysis System in combination with a Zeiss SV8 stereomicroscope at

a magnification of 25x was used to measure amphipods following methods described in Kemble

et al (1994). Amphipods were classified as either "mature male" or "not male" based on the

presence of an enlarged second gnathopod (Kemble et al 1994). An enlarged second gnathopod

of male amphipods was a consistent measure of sexual maturation (it is difficult to distinguish

immature males from females at this age). 
1.5

Chemical and Physical Characterization of Sediments

Acid-volatile Sulfides (AVS) and Simultaneously Extractable Metals (SEM): Subsamples of

sediments were measured for acid-volatile sulfides (AVS) and simultaneously extractable metals

(SEM) immediately after homogenization. Station samples (C samples) were collected on the

boat and stored at 4 C until shipment to the laboratory. Pool samples (B samples) were collected o

in the laboratory immediately after sediment homogenization before the start of toxicity tests. 

Concentrations of AVS in sediment samples were determined using a silver/sulfide electrode

following methods described in Brumbaugh et al (1994). Concentrations of SEM were

determined using atomic spectroscopy following methods described in Brumbaugh et al (1994). 

Percentage recoveries for inorganics from both blank and sediment extracts averaged 96%. 

The average range was from a low of 78% for antimony (spiked as sodium sulfide) in the

sediment extract to a high of 110% for Zn in the sediment extract. The average duplicate

coefficient of variation was 1.7% (6 compounds, n=2). Average duplicate coefficient of

variation ranged from 0.2% for both Pb samples to 5.1% for S in one of the duplicate samples.

Organochlorine Pesticides (OCPs), Polychlorinated Biphenyls (PCBs), and Aliphatic and

Polycyclic Aromatic Hydrocarbons (PAHs): Sediment samples © samples) were prepared for the

analyses of organochlorine pesticides (OCPs), polychlorinated biphenyls (PCBs), and aliphatic

and polycyclic aromatic hydrocarbons (PAHs) by extracting twenty grams of sediment with

acetone, followed by petroleum ether. A final acetone/petroleum ether extraction was done and

the extracts combined, centrifuged and transferred to a separatory funnel containing sufficient

water to facilitate partitioning of residues into petroleum ether portion. The petroleum ether was

washed twice with water and concentrated by Kuderna-Danish to appropriate volume.

Organochlorine determination was conducted by transferring an aliquot of concentrated

extract to a 1.6 g Florisil mini-column topped with 1.6 g sodium sulfate. Residues were eluted

from the column in two elution fractions. The first fraction consisted of 12 mL of hexane

followed by 12 mL of 1% methanol in hexane; the second fraction consisted of an additional 24

mL of 1% methanol in hexane. Quantification of residues in the two Florisil fractions and three

silicic acid fractions was performed using a packed or megabore column and electron capture gas

chromatography.

Hydrocarbon determination was conducted by transferring a second aliquot of the

concentrated extract to a 20 g 1% deactivated silica gel column, topped with 5-g neutral alumina. 

Aliphatic and polynuclear aromatic hydrocarbon residues were fractioned by eluting aliphatics

from the column with 100-mL petroleum ether (Fraction 1) followed by elution of aromatics

using, 100-mL 40% methylene chloride/60% petroleum ether, followed by 50-mL methylene

chloride (combined eluates, Fraction 2). Quantification of fraction 1 by capillary column, flame

ionization gas chromatography was performed once the fraction was concentrated to appropriate

volume. The silica gel (fraction 2) containing aromatic hydrocarbons was concentrated,

reconstituted in methylene chloride and quantified by gas chromatography and mass

spectrometry.

Average percent spike recovery for eighteen OCPs was 103% (n=2). The smallest average

spike recovery was 68% for HCB while o,p’-DDE had the greatest average spike recovery
1.6

(120%). Individual OCP concentrations were below minimum detection limits so duplicate

analyses were not evaluated. Average percent spike recovery for PAH compounds was 98% (29

compounds, n=2). Naphthalene (84%) had the smallest average percent recovery while

fluoranthene had the greatest average spike recovery (110%). The average duplicate coefficient

of variation was 12.6% (13 compounds, n=2). Average duplicate coefficient of variation ranged

from 0% for multiple PAHs in both duplicate samples to 61% for benzo(a)pyrene in one of the

samples.

Methods for the analyses of the B samples, detection limits and quality control are described

in Moody et al (1996). Quality control of B sediment samples analyzed for PAHs included: (1)

estimates of accuracy determined from the standard deviation of the percent recovery of

deuterated compounds added to the extracts and calculated based on absolute area counts and

external calibration, and (2) precision, based on the relative standard deviation of the absolute

area of multiple analyses of a surrogate compound (Moody et al 1996). A list of all the PAHs

and OCPs analyzed for in both sets of sediment samples (B and C) are listed in Appendix 1.3. 

Physical Characterization of Sediments

Physical characterization of sediments included: (1) percentage water (Kemble et al 1993), (2)

particle size using a hydrometer (Forth et al 1982; Gee and Bauder 1986; Kemble et al 1993),

and (3) total organic carbon using a coulometric titration (Cahill et al 1987; Kemble et al 1993). 

All physical characterizations included analysis of duplicate samples. Differences in percentage

water for duplicate samples ranged from 0% in treatments 2B, 7B, 13C, 14B and 18B to 7% in

treatment 10C. Duplicate samples of control sediment, sucrose standards and blanks were

analyzed when determining sediment total organic. Precision and accuracy of the coulometric

technique used was tested against National Bureau of Standards and Standard Reference

Materials (NBS-SRM) with an error of less than 0.03% of the excepted values (Cahill et al

1987). Differences between duplicates ranged from 0% in treatments 3B, 11B, 12B, 13C, 14C,

15C, 18C, 20C, 22C, 22B, 24C and 26C to 0.9% in treatments 5C, 9C and 26B.

Data Analysis and Statistics

Toxicity Tests: Before statistical analyses were performed, data for survival and maturation were

arcsin transformed. Comparisons of mean survival and percentage sexual maturation were made

using a one-way nested analysis of variance (ANOVA) with mean separation by Fisher’s

protected least significant difference test at alpha = 0.05 (Snedecor and Cochran 1982). Data for

length had a normal distribution and were not transformed before statistical analysis. 

Comparison of mean body length was made using a one-way ANOVA with mean separation by

Fisher’s protected least significant difference test at alpha = 0.05 (Snedecor and Cochran 1982). 

A sample was designated as toxic when survival, growth, or sexual maturation were significantly

reduced relative to the control and reference sediments. Sediments from pools 6 and 11 were

chosen as reference sediment based on low concentrations of contaminants. Simple linear

regression was used to compare physical and chemical sediment characteristics to amphipod

survival, length or sexual maturation. All statistical analyses were performed with Statistical
1.7

Analysis System (SAS) programs (SAS 1994).

Effects Range Median: Chemistry concentrations and toxicity endpoints were evaluated using

28-day Hyalella azteca Effect Range Medians (ERMs) reported by Ingersoll et al (1996) and

Smith et al (1996). An ERM is defined as the concentration of a chemical in sediment above

which effects are frequently or always observed or predicted for most species (Long et al 1995). 

The total number of individual ERMs exceeded with each sample was plotted against the sum

ERM quotient (SERM-Q; where Q is equal to the concentration of each chemical in the sediment

sample divided by the ERM for that chemical), similar to the toxic unit described by Canfield et

al (1996), Ingersoll et al (1996) and Swartz et al (1997). We chose to evaluate sediment toxicity

relative to nine ERMs which correctly classified >70% of the samples in Ingersoll et al (1996). 

These 9 individual ERMs tended to minimize Type I (false positive) and Type II (false negative)

errors relative to other SECs reported by Ingersoll et al (1996). Due to insufficient chemistry

data for chromium and total PCBs, only 7 of the 9 individual ERMs were used in this evaluation. 

These ERMs included: cadmium, lead, nickel, zinc, chrysene, benzo(a)pyrene, and

benzo(g,h,i)perylene.

Results and Discussion

Toxicity Tests

Survival of amphipods was significantly reduced relative to the control and reference sediments

only in the 13B treatment (Table 1.1). Body length of amphipods was significantly reduced

relative to the control and reference sediments in only the 26C treatment (Table 1.1; Appendices

1.4 and 1.5). Sexual maturation was not significantly reduced in any treatments when compared

to the control and reference sediments (Table 1.1; Appendices 1.6 and 1.7).

Indigenous organisms recovered at the end of amphipod exposures included oligochaetes,

ostracods, clams, and a snail. Clam shells were present in many of the sediments; however, only

a few live clams were retrieved at the end of the exposure. Pairs of amphipods were observed in

amplexus in the control, 1-B, 2-B, 5-B, 6-C, 8-B, 8-C, 9-B, 10-B, 11-B, 14-C, 15-B, 18-C, 24-B,

24-C, and 26-B treatments, and gravid females were observed in the control, 11-B, 16-C, and 24-

B treatments. 

Although significant differences in survival of amphipods relative to the control and reference

sediments were only observed in sample 13B, there was a relatively wide range in survival

among the treatments. For example survival was below 70% in 13 of the 51 treatments (Table

1.1). Survival of amphipods in the control was acceptable ($80%), however, survival in two of

the four reference treatments (11C and 6B) was below 80%. Subsequent studies have found that

the reconstituted water described in USEPA (1994) that was used to conduct this study does not

consistently support adequate survival and growth of Hyalella azteca in 28-day exposures

(McNulty 1995; Kemble et al 1996). Ingersoll et al (1997) retested sediment samples 4C, 11C,

14C, and 24C using well water as an overlying water and observed a mean survival of >90% in

all of the samples with no substantial effects on growth, or reproduction of H. azteca. Survival

of amphipods in these same sediments ranged from 48% to 63% in the present chapter (Table
1.8

1.1). Similarly, Benoit et al (1997) tested Station samples (7C, 9C, 13C, 22C, and 24C) in

chronic toxicity tests with midge Chironomus tentans using a natural overlying water and did not

observe effects on survival, growth, emergence, or reproduction. Additional studies are ongoing

to evaluate 28-day Hyalella azteca exposures using reconstituted waters.

Physical and Chemical Characteristics of Sediments

Physical and chemical characteristics of sediment samples are listed in Table 1.2. Sediment

organic carbon content ranged from 0.2% for the sediment samples from Stations 6B and 20B to

5.2% for Station 10C. Organic carbon content in the control sediment was 1.2%. Percentage

solids ranged from 21% in the sediment sample from stations 4C and 10C to 84% for the

sediment sample from Station 20B. Classification of the sediment samples for grain size varied

from pool to pool (i.e., loam (11C), sandy-loam (8B), silty-clay-loam (25 C and 22C)) while the

control sediment was a silty-clay-loam (Table 1.2). Acid volatile sulfide levels ranged from

0.005 μmoles/g in the 1C sample to 63.0 μmoles/g in the 10C sample (Table 1.2).

Concentrations of simultaneously extracted metals in sediment samples are listed in Appendix

1.8. Sediment from sample 4C had the highest concentrations of extractable SEM Cd, Cu, Ni,

and Pb. Sample 12C had the highest concentration of SEM Zn (Appendix 1.8). The sum

SEM/AVS molar ratio in the present study was typically less than 1 (except the two samples

from pool 1). This indicates the concentration of divalent metals listed in Appendix 1.8 were

probably not high enough to result in toxicity of the samples (DiToro et al 1990). Concentrations

of SEM Cd, Cu, Ni and Pb were highest in sediment samples from treatment 4C (Appendix 1.8). 

However, concentrations of SEM Cu and Pb were still below the ERMs reported by Ingersoll et

al (1996; Figures 1.2 and 1.3). 

Significant positive correlations were observed between SEM metals vs. TOC (Cu > Zn >

Cd>Pb>Ni), SEM metals vs. percentage clay (Zn>Ni>Pb>Cu>Cd) and between SEM metals vs.

percentage silt (Ni>Cu>Pb>Zn>Cd) when tested by Spearman's rho coefficient of rank

correlation (Table 1.3). The significant negative correlation with sand and the positive

correlation with clay and silt indicates that metals were concentrated in the finer sediment

particles.

Concentrations of organochlorine pesticides (OCPs) in sediment samples are listed in

Appendix 1.9. Concentrations of OCPs were below detection limits (0.01 μg/g) in all of the C

samples except the 2C and SCC samples which had detectable concentrations of DDE and DDD

(Appendix 1.9). Amphipod survival in the 2C sediment sample was 75%. However, despite

having concentrations which were similar for both chemicals, survival of amphipods in the SCC

sample was 90%. This indicates that the levels of DDE and DDD detected in these samples was

not the sole cause of the lower survival observed in the 2C sediment sample. Concentrations of

OCPs in the B samples were at or below detection limits for 10 of the 15 individual pesticides

evaluated (Appendix 1.9). Concentrations for all 5 OCPs detected in the B samples were #0.079

μg/g dry weight and were below calculated ERMs (Smith et al 1996; Appendix 1.9).

Concentrations of polycyclic aromatic hydrocarbons (PAHs) in sediment samples are listed in

Appendix 1.10. The highest concentrations were observed at Pool 1 and were generally lower in

the downstream pools. Concentrations of PAHs in river sediments exceeded the Method Lower
1.9

Limit of Quantitation (MLLQ; 0.03 μg/g) in at least one sediment sample for every PAH

evaluated (except for 1-methylnaphthalene; Appendix 1.10). Concentrations of 4 of the 11 PAHs

measured exceed at least one calculated ERM (Ingersoll et al 1996; Figures 1.4 and 1.5). 

Elevated PAH concentrations in sediment samples were associated with sediment collected from

pools near Minneapolis, MN. Concentrations of PAHs below pool 4 were similar in the

remaining pools. Concentrations of fluoranthene exceeded the calculated ERM (0.175 μg/g) in 9

of the sediment samples from the Upper Mississippi River. Amphipod survival in these samples

was above 75% in all but one of the samples (sample 4C which had a survival of 63%; Table

1.1). This would indicate that concentrations of fluoranthene in these samples had little or no

effect on amphipod survival.

Comparisons of Sediment Characteristics to Toxicity Responses

Relationships of physical or chemical characteristics of sediments to toxicity were evaluated

using rank correlation (Table 1.4). No significant correlations were observed between survival,

growth or maturation and the measured physical or chemical characteristics of the sediment

samples (Table 1.4). Additionally, no significant correlation was observed between the toxicity

endpoints and concentrations of PAHs or OCPs normalized to total organic carbon

concentrations (Table 1.5). Sediments from Pool 1 had the highest percent sand (>88%), but

amphipod length and maturation were not reduced with exposure to 1B or 1C sediments relative

to the control and reference sediments (Table 1.1). Similarly, the control sediment had the

highest percent silt and clay relative to the other samples. Ingersoll and Nelson (1990), Kemble

et al (1994), and Ingersoll et al (1997) also reported sediment particle size did not affect the

response of Hyalella azteca in 28-d sediment exposures.

None of the 49 sediment samples exceeded any of the 7 individual ERMs. Use of these 7

ERMs correctly classified 47 of the 49 (96%) sediment samples from the UMR as non-toxic. 

The two samples incorrectly classified were both type II errors (false negative; toxic sample that

does not exceed an ERM). This again may indicate something other than contaminants or

contaminants not measured were the cause of the relatively wide range in survival among the

treatments.

Additional ERMs for individual chemicals listed in Ingersoll et al (1996) and Smith et al

(1996) were also evaluated. About 20% of the sediment samples exceeded at least one of these

ERMs. However, use of these additional ERMs to classify samples as toxic or non-toxic resulted

in increased Type I error (false positive; non-toxic sample that exceeds an ERM). As was the

case when using only the seven ERMs, chemical concentrations from the two samples classified

as toxic did not exceed any of the additional ERMs.

The prediction of sediment toxicity was also evaluated using a toxic quotient approach. A

toxic quotient was calculated for each sample by first dividing the concentration of individual

chemicals by their respective ERM and then summing each of the individual values (Canfield et

al 1996; Ingersoll et al. 1996). In the present study, quotients for the seven chemicals listed

above were used to calculate a toxic quotient for each sample (Table 1.2). Figure 1.6 plots the

relationship between the frequency of ERM exceedances and the sum of the ERM toxic quotient. 

In the present study, the ERM toxic quotient was # 2.6 and individual ERMs were not exceeded
1.10

indicating the sediment samples from the UMR were relatively non-contaminated compared to

sediments from areas of known contamination in the United States (Kemble et al 1994; Ingersoll

et al 1996). A toxic quotient approach was also used in Chapter 3 using a quadrant frequency

analysis to evaluate the benthic community of the pools in the UMR system.

Summary

Toxicity tests using amphipods identified only two of the 49 sediment samples from the Upper

Mississippi River system as toxic (a significant reduction in survival, growth or sexual

maturation compared to the control and reference sediments). However, there was a relatively

wide range in survival among the treatments. The overlying water used in this test was the

reconstituted water described in USEPA (1994), which McNulty (1995) and Kemble et al (1996)

have reported does not consistently support adequate survival of Hyalella azteca in 28-d

sediment exposures. Survival of amphipods and midge was >90% in subsequent studies with

sediments from the present study when natural water was used as the overlying test water (Benoit

et al 1997; Ingersoll et al 1997). This would indicate that the reconstituted test water was a

significant factor in the wide range of survival observed in the present study.

Effect Range Medians (ERMs) were used to evaluate the toxicity of contaminants associated

with field collected sediments. ERMs correctly classified 96% of the UMR sediment samples as

non-toxic. The two samples incorrectly classified were type II errors (false negatives). Again

this indicates that factors other than contaminants or unmeasured contaminants may have been

responsible for the variation in amphipod survival that was observed.

Concentrations of contaminants in sediments from the UMR were typically 10 to 100 times

less than concentrations of contaminants in sediments previously associated with toxicity

(Kemble et al 1994; Ingersoll et al 1996; Figure 1.7). This would indicate that the sediment

samples from the UMR were relatively non-contaminated compared to other areas of know

contamination across the United States.

Acknowledgment. We would like to thank Linda Sappington for assistance on Quality Control

and Quality Assurance and the following individuals for their input on the project: Eugene Greer,

Doug Hardesty, Pam Haverland, Chris Henke, Ed Henry, Phil Lovely, John Moody, Shane

Reussler, Julie Soltvedt, Jeff Stevens, Ron Walton, Dave Whites, Dave Zumwalt, the Crew of the

Acadiana (Craig LeBoeuf and Pat Marmande), and the laboratories providing chemical analysis

of the sediment samples. Thanks to Dave Mount, James Fairchild and Parley Winger and two

anonymous reviewers for their helpful comments on the manuscript.
1.11

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Fig. 1.1. Map of the Upper Mississippi River (UMR) from Minneapolis, MN to Saint Louis, MO.
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B SAMPLESC SAMPLESERM

1.16

Fig. 1.2. Concentrations of Simultaneously Extracted Metal (SEM) Cd in UMR sediment samples compared to a 
1

2

3

4

5

6

7

8

9

10

11

12

13

14

15

16

18

1

Pool

0

20406080

100120

Pb (ug/g) 

B SAMPLESC SAMPLESERM

1.17

Fig. 1.3. Concentrations of SEM Pb in UMR sediment samples compared to a ERM for Pb.
1

SC

2

3

4

5

6

7

8

9

10

11

12

13

14

15

16

1

Pool

0000011

Chrysene (ug/g)

B SamplesC SamplesERM

1.18

Fig. 1.4. Concentrations of Chrysene in UMR sediment samples compared to a ERM for Chrysene.
1

SC

2

3

4

5

6

7

8

9

10

11

12

13

14

15

16

18

Pool

000001

Benzo(a)pyrene (ug/g)

B SamplesC SamplesERM

1.19

Fig. 1.5. Concentrations of Benzo(a)pyrene (BAP) in UMR sediment samples compared to a ERM for BAP.
0

0

1

10

SUM ERM TOXIC QUOTIENT 

01234567

ERM EXCEEDANCES (#)other non-toxic other toxic umr non-toxic umr toxic

1.20

Fig. 1.6: Number of ERM exceedances for the 7 chemicals that correctly classified 70% of the samples compare
1

10

100

SUM ERM TOXIC QUOTIENT

0

20406080

100

SURVIVAL (%) 

1.21

Fig. 1.7. Survival vs Sum ERM toxic quotients of sediment samples from the UMR compared to survival vs. sum and the Clark Fork River and Milltown Reservoir MT.
1.22

Table 1.1. Results of the Upper Mississippi River sediment tests with Hyalella azteca. Means (Standard error of

the means in parentheses) within a column and within a set of sample are significantly different (p <0.05; n=4) from

the control and reference sediment and are designated with an asterix.

 

Sample Survival (%) Length (mm) Mature Males (%) 1

 

1st set of samples

Control 80.0 (4.08) 3.39 (0.16) 36.7 (8.91)

1B 92.5 (4.79) 3.66 (0.11) 39.1 (5.71) 

1C 65.0 (5.00) 3.17 (0.11) 16.9 (6.90)

3B 95.0 (5.00) 4.27 (0.08) 44.9 (8.43)

5B 80.0 (7.07) 4.23 (0.06) 44.8 (10.30) 

5C 80.0 (7.07) 4.06 (0.10) 21.6 (4.23)

8B 97.5 (2.50) 3.69 (0.09) 40.5 (7.72)

8C 92.5 (2.50) 4.09 (0.11) 32.3 (7.68)

10B 92.5 (7.50) 4.28 (0.09) 39.5 (18.49)

10C 72.5 (13.15) 3.86 (0.08) 34.4 (6.88)

11B (reference) 87.5 (2.50) 4.31 (0.07) 43.3 (11.57)

11C (reference) 57.5 (8.54) 3.61 (0.07) 32.8 (15.79)

12B 72.5 (9.46) 3.48 (0.07) 34.5 (3.00)

12C 85.0 (6.45) 3.78 (0.07) 32.4 (5.85)

15B 90.0 (4.08) 3.74 (0.08) 51.3 (11.46)

15C 72.5 (2.50) 3.59 (0.09) 34.0 (8.64)

16B 70.0 (9.13) 3.72 (0.08) 40.6 (6.56)

16C 90.0 (7.07) 3.83 (0.07) 30.0 (10.13)

21B 95.0 (2.89) 3.46 (0.06) 52.2 (6.08)

21C 87.5 (4.79) 3.87 (0.09) 51.4 (5.29)

25B 62.5 (13.15) 3.60 (0.11) 23.8 (10.51)

25C 62.5 (15.48) 3.63 (0.08) 29.6 (8.34)

26B 92.5 (4.79) 3.51 (0.09) 42.0 (6.82)

26C 90.0 (7.07) 2.88 (0.01) * 48.8 (11.30) 

2nd Set of samples

Control 97.5 (2.50) 2.59 (0.08) 5.9 (3.42)

2B 75.0 (8.66) 4.07 (0.11) 31.3 (6.25)

2C 75.0 (10.41) 3.47 (0.10) 43.8 (8.08)

4B 85.0 (6.45) 3.39 (0.10) 36.7 (13.72)

4C 62.5 (21.75) 3.35 (0.09) 12.1 (5.22)

6B (reference) 67.5 (17.02) 3.53 (0.09) 26.9 (9.21) 

6C (reference) 82.5 (2.50) 4.08 (0.10) 54.5 (2.97)

7B 100.0 (0.00) 3.66 (0.06) 42.5 (10.31)

7C 95.0 (2.89) 3.70 (0.07) 35.5 (3.41) 

9B 75.0 (10.41) 3.72 (0.09) 43.6 (6.47)

9C 67.5 (13.77) 3.65 (0.08) 32.8 (11.24)

13B 32.5 (7.50) * 3.87 (0.19) 18.8 (11.97)

13C 47.5 (10.31) 3.56 (0.11) 50.0 (9.64)

14B 65.0 (5.00) 3.85 (0.12) 31.6 (7.36)
1.23

Table 1.1. (continued)

 

Sample Survival (%) Length (mm) Mature Males (%) 1

 

14C 47.5 (7.50) 3.50 (0.12) 43.8 (15.72)

18B 77.5 (7.50) 3.57 (0.12) 50.0 (18.89)

18C 72.5 (17.97) 3.52 (0.09) 20.8 (7.50)

19B 85.0 (6.45) 3.31 (0.07) 40.2 (7.50)

19C 72.5 (7.50) 3.44 (0.07) 32.3 (15.91)

20B 82.5 (8.54) 3.43 (0.08) 11.9 (5.14)

20C 95.0 (2.89) 3.30 (0.06) 27.2 (10.74)

22B 85.0 (6.45) 3.79 (0.10) 24.4 (3.00)

22C 52.5 (10.31) 3.64 (0.11) 39.9 (14.20)

24B 87.5 (2.50) 3.61 (0.08) 34.4 (4.65)

24C 60.0 (8.16) 3.78 (0.12) 66.9 (14.19)

SCB 75.0 (10.41) 3.42 (0.10) 11.9 (7.89)

SCC 90.0 (4.08) 3.03 (0.06) 31.7 (5.60) 

starting body length of amphipods in the 1st set of samples was 1.05 mm (0.02 SE, n=11) and was 1.17 mm (0.04 1

SE, n=10) in the 2nd set of samples.
1.24

Table 1.2. Physical and chemical characteristics of sediments from the Upper Mississippi River at the start of whole-sediment tests. The sum Ealso calculated for each sample.

 TotalOrganic Particle Size (%) Sum

Sample Carbon Solids sand clay silt ERM

 (%) (%) Quotient Sediment Class

1B 0.3 76.5 88.6 9.3 2.1 1.17 Sand/Loamy Sand1C 0.5 77.9 88.8 10.1 1.1 0.80 Sand/Loamy Sand2B 3.6 61.3 53.5 25.5 21.0 1.66 Sandy Clay Loam2C 3.3 45.0 15.4 43.1 41.5 1.58 Silty Clay3B 2.7 53.2 27.5 23.5 49.0 1.00 Loam4B 4.8 26.2 11.6 49.0 39.5 1.68 Clay4C 5.0 20.8 33.4 39.8 26.9 2.60 Clay Loam5B 1.6 61.5 53.6 19.4 26.4 0.22 Sandy Loam5C 5.1 27.7 31.6 31.0 37.5 0.81 Clay Loam6B 0.2 77.3 84.6 12.4 3.0 0.11 Loamy Sand6C 0.7 70.2 78.1 13.6 8.3 0.28 Sandy Loam7B 1.0 47.7 17.1 32.1 50.7 0.27 Silty Clay Loam7C 2.3 62.1 56.5 16.8 26.7 0.65 Sandy Loam8B 1.3 57.5 58.0 18.8 23.2 0.21 Sandy Loam8C 2.2 55.5 11.5 37.0 51.5 0.47 Silty Clay Loam9B 2.0 56.3 27.6 21.5 50.9 0.52 Silt Loam9C 2.9 48.0 9.3 29.4 61.3 0.60 Silty Clay Loam10B 1.2 55.2 59.6 36.9 3.5 0.25 Sandy Clay10C 5.2 20.7 24.3 41.7 34.0 0.94 Clay11B 1.3 59.8 46.1 18.8 35.1 0.28 Loam11C 1.8 64.7 46.2 21.6 31.3 0.31 Loam12B 2.0 54.2 20.0 20.9 59.1 0.77 Silt Loam12C 2.3 54.9 15.3 21.4 63.3 0.84 Silt Loam13B 1.8 65.4 33.2 23.1 43.7 0.24 Loam13C 1.8 52.1 14.6 22.0 63.4 0.50 Silt Loam14B 0.6 35.8 4.0 42.5 53.5 0.23 Silty Clay14C 3.0 61.0 58.7 18.4 22.9 0.70 Sandy Loam15B 1.4 46.9 0.0 23.0 77.0 0.48 Silt Loam15C 1.9 59.0 41.5 20.5 38.0 0.59 Loam
1.25

Table 1.2. (continued).

 TotalOrganic Particle Size (%) Sum

Sample Carbon Solids sand clay silt ERM

 (%) (%) Quotient Sediment Class

16B 1.2 67.0 53.7 18.9 27.4 0.39 Sandy Loam16C 2.8 67.4 51.3 21.9 26.8 0.76 Sandy Clay Loam18B 0.7 69.1 64.0 19.5 16.5 0.33 Sandy Loam18C 1.7 62.6 21.8 23.8 54.5 0.56 Silt Loam19B 1.9 54.9 33.8 29.4 36.9 0.47 Clay Loam19C 2.3 49.2 7.6 34.0 58.4 0.66 Silty Clay Loam20B 0.2 84.1 81.4 11.7 6.8 0.23 Loamy Sand20C 0.8 73.5 52.1 22.0 26.0 0.45 Sandy Clay Loam21B 0.5 69.9 64.0 23.5 12.5 0.32 Sandy Clay Loam21C 1.1 59.0 44.4 25.8 29.8 0.30 Loam22B 0.5 73.3 62.1 23.4 14.5 0.25 Sandy Clay Loam22C 2.4 44.4 0.3 40.3 59.4 0.59 Silt Clay Loam24B 0.7 74.6 57.5 23.0 19.5 0.21 Sandy Clay Loam24C 1.7 57.1 30.7 22.0 47.4 0.50 Loam25B 1.4 63.3 33.2 30.7 36.1 0.33 Clay Loam25C 1.1 56.2 16.6 28.0 55.4 0.38 Silty Clay Loam26B 2.0 54.5 24.1 33.5 42.5 0.51 Clay Loam26C 0.7 72.6 43.5 27.0 29.5 0.42 Clay LoamSCB 3.0 34.0 53.4 24.8 21.9 0.88 Sandy Clay LoamSCC 4.3 26.6 36.1 25.5 38.5 1.17 LoamFLORB 1.2 32.0 12.3 26.5 61.3 1.04 Silty Clay LoamFLORC 1.2 32.0 12.3 26.5 61.3 1.04 Silty Clay Loam
1.26

Table 1.3. Spearman rank correlation for SEM Cd, Cu, Ni, Pb, and Zn with TOC, percent Sand, percent Silt, and percen

River sediments (excluding the control sediment). All of the correlations listed below were significant (p # 0.05).

Element TOC% %Sand %Silt %ClayCd 0.826 -0.449 0.341 0.394Cu 0.868 -0.556 0.563 0.468Ni 0.808 -0.634 0.594 0.553Pb 0.823 -0.583 0.434 0.549Zn 0.854 -0.589 0.385 0.570
1.27

Table 1.4. Linear regression (r ) of amphipod survival, length, or sexual maturation to sediment physical and 2

chemical characteristics. None of the regression were significant (p<0.05).

 Sexual 

 Survival Length Maturation

PW Total Ammonia 0.11 0.07 0.17

PW Unionized Ammonia 0.01 0.06 0.03

PW Total Sulfide 0.05 0.05 0.04

PW Hydrogen Sulfide < 0.01 0.03 0.09

PW Alkalinity < 0.01 0.09 0.08

PW Hardness 0.01 0.16 0.13

PW pH 0.02 < 0.01 < 0.01

PW DO < 0.01 0.04 0.01

PW conductivity 0.01 0.13 0.03

AVS 0.11 < 0.01 0.02

Total Organic Carbon 0.02 < 0.01 0.02

Percent Sand 0.02 < 0.01 < 0.01

Percent Clay 0.01 < 0.01 0.01

Percent Silt 0.05 < 0.01 0.05

Percent Fines 0.02 < 0.01 < 0.01 1

Percent Water < 0.01 0.02 0.02

SEM Cd 0.03 0.04 0.11

SEM Cu 0.01 0.03 0.03

SEM Ni < 0.01 < 0.01 < 0.01

SEM Pb 0.02 0.05 0.05

SEM Zn 0.02 0.01 0.02

Toxaphene 0.01 0.02 < 0.01

Mirex 0.05 0.04 < 0.01

DDD < 0.01 0.10 < 0.01

DDT 0.05 0.04 < 0.01

DDE < 0.01 0.12 0.00

Endrin 0.05 0.04 < 0.01

Dieldrin 0.05 0.04 < 0.01

Heptachlor epoxide 0.05 0.04 < 0.01

Lindane 0.05 0.04 < 0.01

Naphthalene 0.04 0.04 0.01

Acenaphthalene 0.04 < 0.01 < 0.01

Acenaphthene < 0.00 0.02 0.07

Phenanthrene 0.02 0.01 0.01

Anthracene < 0.01 < 0.01 < 0.01
1.28

Table 1.4. (Continued)

 Sexual 

 Survival Length Maturation

Fluorene 0.01 0.01 < 0.01

Fluoranthene 0.04 < 0.01 < 0.01

Chrysene 0.02 < 0.01 < 0.01

Pyrene 0.03 < 0.01 < 0.01

Benzo(b)fluoranthene 0.04 < 0.01 < 0.01

Benzo(k)fluoranthene 0.01 0.07 0.03

Benzo(a)pyrene 0.01 0.05 0.04

Indeo(1,2,3,-cd)pyrene < 0.01 < 0.01 0.01

Benzo(g,h,j,i)perylene < 0.01 0.01 0.01

Silt and Clay combined 1
1.29

Table 1.5. Linear regression (r ) of amphipod survival, length, sexual maturation to sediment chemical 2

characteristics normalized to organic carbon. None of the regressions were significant (p<0.05).

 Sexual 

 Survival Length Maturation

Toxaphene 0.03 0.01 0.05

Mirex 0.02 0.04 < 0.01

DDD 0.01 0.10 0.01

DDT 0.01 0.02 0.01

DDE < 0.01 0.03 < 0.01

Endrin 0.01 0.05 < 0.01

Dieldrin < 0.01 0.07 < 0.01

Heptachlor epoxide 0.01 0.07 < 0.01

Lindane 0.02 0.05 < 0.01

Naphthalene 0.03 < 0.01 < 0.01

Acenaphthalene < 0.01 < 0.01 < 0.01

Acenaphthene 0.02 0.02 0.02

Phenanthrene 0.01 < 0.01 < 0.01

Anthracene < 0.01 0.06 < 0.01

Fluorene < 0.01 < 0.01 0.01

Fluoranthene 0.04 < 0.01 < 0.01

Chrysene 0.14 0.07 < 0.01

Pyrene 0.05 0.06 < 0.01

Benzo(b)fluoranthene 0.06 0.02 < 0.01

Benzo(k)fluoranthene 0.01 0.03 < 0.01

Benzo(a)pyrene 0.04 0.07 < 0.01

Indeo(1,2,3,-cd)pyrene < 0.01 0.05 0.01

Benzo(g,h,j,i)perylene < 0.01 0.05 0.01
1.30

Appendix 1.1. Pore water quality for the whole-sediment tests with Upper Mississippi river samples. 

Total unionized Total 

Pool pH Alkalinity Hardness DO Conductivity ammonia ammonia Sulfide 

(mg/L) (mg/L) (mg/L) (μmho @25 C) (mg/L) (mg/L) (mg/L) 

o

01B 7.33 376 852 4.60 758 7.240 0.008 0.019 01C 7.69 292 280 7.60 541 1.540 0.004 0.000 02B 7.38 732 758 2.38 1354 5.900 0.007 0.011 02C 7.47 560 552 4.80 969 3.310 0.005 0.005 03B 7.50 624 664 6.60 1131 3.980 0.006 0.006 04B 7.59 ND ND 4.23 779 2.745 0.005 0.302 04C 7.84 ND ND 7.90 747 1.370 0.005 ND 05B 7.61 455 460 4.30 832 3.310 0.007 ND 05C 7.40 374 360 3.50 700 3.580 0.004 0.011 06B 7.82 432 404 6.20 830 3.470 0.011 0.099 06C 7.46 548 580 5.45 1031 11.400 0.016 0.040 07B 7.50 636 652 1.50 1186 5.640 0.009 0.000 07C 7.29 396 ND 4.35 765 6.150 0.006 0.009 08B 7.62 522 519 5.10 961 3.420 0.007 0.000 08C 7.45 596 600 5.60 1055 6.480 0.009 0.011 09B 7.43 835 750 6.15 1462 10.500 0.014 0.037 09C 7.41 ND ND 4.00 1262 10.400 0.013 0.124 10B 7.48 488 480 4.05 912 4.730 0.007 0.002 10C 7.26 452 468 5.45 785 3.550 0.003 0.039 11B 7.50 ND ND 5.60 930 4.120 0.006 0.000 11C 7.45 415 418 5.90 786 4.440 0.006 0.007 12B 7.40 568 620 6.00 1013 4.900 0.006 0.037 12C 7.20 710 600 4.60 1163 6.350 0.005 0.018 13B 7.24 852 808 4.60 1680 22.700 0.020 0.013 13C 7.41 ND ND 4.75 897 8.070 0.010 0.011 14B 7.43 436 440 2.55 846 4.540 0.006 0.029 14C 7.53 ND ND 4.35 636 4.290 0.007 0.465 15B 8.17 ND ND 5.80 847 3.440 0.025 0.023 15C 7.47 364 360 7.20 671 2.360 0.003 0.012 16B 7.50 464 484 3.50 892 6.190 0.010 0.003 16C 7.40 ND ND 5.40 998 6.970 0.009 0.031 18B 7.37 420 408 5.30 835 4.690 0.005 0.025 
1.31

Appendix 1.1. Pore water quality for the whole-sediment tests with Upper Mississippi river samples (continued).18C 7.44 340 348 2.95 652 3.180 0.004 0.007 19B 7.49 573 505 3.00 1027 5.440 0.008 ND 19C 7.33 ND ND 3.90 1077 6.840 0.007 0.202 20B 0.00 ND ND ND ND 2.750 0.000 0.569 20C 7.78 ND ND 6.80 643 2.650 0.008 0.007 21B 7.45 ND ND 5.30 1019 9.030 0.013 0.001 21C 7.31 ND 540 2.50 945 8.730 0.009 0.041 22B 7.41 495 475 5.10 934 6.370 0.008 0.000 22C 7.20 ND ND 4.50 1109 12.300 0.010 0.159 24B 7.65 ND ND 6.75 568 2.780 0.006 0.003 24C 7.45 ND ND 6.40 708 ND 0.000 0.052 25B 7.34 ND ND 1.50 869 3.260 0.004 ND 25C 7.42 440 352 5.60 768 2.270 0.003 0.008 26B 7.47 528 528 4.95 1003 5.520 0.008 0.007 26C 7.45 480 484 5.70 891 3.800 0.005 0.001 SCB 7.27 244 216 6.95 380 1.370 0.001 0.038 SCC 7.19 ND 148 6.50 386 1.210 0.001 0.057 FLOR 6.69 ND ND 9.35 1176 1.410 0.000 0.001 Mean 7.45 505 504 5.04 906 5.320 0.007 0.055 Max. 8.17 852 852 9.35 1680 22.700 0.025 0.569 Min. 6.69 244 148 1.50 380 1.210 0.000 0.000 Std 0.21 141 159 1.60 246 3.649 0.005 0.114 Median 7.45 480 484 5.10 892 4.440 0.007 0.012 
1.32

Appendix 1.2. Mean measured overlying water quality for the whole-sediment tests with Upper Mississippi river samples. Water quality was co

27.

 Total unionized

Pool pH Alkalinity Hardness DO Conductivity ammonia ammonia

(mg/L) (mg/L) (mg/L) (μmho @25 C) (mg/L) (mg/L) o

01B 7.77 78 115 7.20 375 0.303 0.00101C 7.87 72 112 7.44 367 0.045 0.00002B 8.14 102 139 6.32 419 0.634 0.00402C 8.72 151 160 6.52 408 0.473 0.01203B 8.16 84 128 6.50 402 0.190 0.00104B 7.96 83 129 6.04 396 0.312 0.00104C 8.09 76 125 7.04 386 0.171 0.00105B 8.10 78 121 7.18 388 0.183 0.00105C 8.36 79 122 7.20 363 0.308 0.00406B 7.97 81 130 7.02 391 0.331 0.00206C 8.01 122 124 6.05 396 0.981 0.00507B 8.10 115 144 7.06 391 0.610 0.00407C 8.00 80 126 6.80 378 0.684 0.00308B 7.95 79 120 7.04 386 0.218 0.00108C 8.04 88 127 6.36 400 0.453 0.00209B 8.17 111 148 6.44 413 0.902 0.00709C 8.08 93 148 6.40 402 0.857 0.00510B 8.15 82 122 7.04 396 0.285 0.00210C 8.01 83 124 6.86 392 0.268 0.00111B 8.11 78 119 6.94 386 0.310 0.00211C 8.16 75 120 7.02 398 0.228 0.00212B 8.30 83 125 7.05 395 0.228 0.00212C 8.28 84 125 6.74 394 0.292 0.00313B 8.06 129 154 5.84 428 1.520 0.00913C 8.09 101 135 6.24 398 0.808 0.00514B 8.02 94 135 6.21 403 0.570 0.00314C 8.09 87 136 6.24 403 0.397 0.00215B 8.20 78 117 6.88 400 0.160 0.00115C 7.92 80 122 6.50 385 0.157 0.00116B 8.15 81 126 6.76 412 0.324 0.00216C 8.18 85 127 6.72 399 0.350 0.003
1.33

Appendix 1.2. Mean measured overlying water quality for the whole-sediment tests with Upper Mississippi river samples (continued).18B 7.97 86 129 6.00 399 0.480 0.00218C 8.19 83 144 6.40 391 0.354 0.00319B 8.10 98 140 6.26 411 0.842 0.00519C 8.07 94 140 5.92 411 0.593 0.00320B 8.08 79 127 7.06 388 0.090 0.00120C 7.97 78 119 7.02 374 0.369 0.00221B 8.14 85 113 7.04 387 0.346 0.00221C 8.10 83 124 6.26 395 0.423 0.00322B 8.13 104 141 6.37 404 1.214 0.00822C 8.09 94 140 6.24 410 0.587 0.00424B 8.15 87 133 7.08 381 0.194 0.00124C 8.14 96 136 6.76 397 0.377 0.00325B 8.04 74 114 7.02 388 0.129 0.00125C 8.05 76 116 6.94 385 0.220 0.00126B 8.06 78 120 6.50 394 0.511 0.00326C 8.03 76 121 6.54 383 0.254 0.001SCB 7.74 60 113 6.88 364 0.199 0.001SCC 7.86 66 111 7.04 359 0.199 0.001FLOR B7.71 59 115 7.53 373 0.220 0.001FLOR C7.58 71 116 7.24 369 0.091 0.000Mean 8.07 87 128 6.70 392 0.416 0.003Max. 8.72 151 160 7.53 428 1.520 0.012Min. 7.58 59 111 5.84 359 0.090 0.000Std 0.17 16 11 0.41 14 0.294 0.002Median 8.09 83 125 6.76 394 0.324 0.002
1.34

Appendix 1.3. List of polycyclic aromatic hydrocarbons (PAHs) and organochlorines (OCs) analyzed for in the sediment samples

from the Upper Mississippi River.

Polycyclic aromatic hydrocarbons

1. Naphthalene 23. 2-methylnaphthalene

2. 1-methylnaphthalene 24. Biphenyl

3. 2,6-dimethylnaphthalene 25. Acenaphthalene

4. Acenaphthene 26. 2,3,5-trimethylnaphthalene

5. Fluorene 27. Dibenzothiophene

6. Phenathrene 28. Anthracene

7. 1,-methylphenanthrene 29. Fluoranthene

8. Pyrene 30. Benzo(b)fluoranthene

9. Chrysene 31. Benzo(k)fluoranthene

10. 1,2-Benzanthracene 32. Benzo(e)pyrene

11. Perylene 33. Benzo(a)pyrene

12. Indeno(1,2,3-cd)pyrene 34. 1,2,5,6-dibenzanthracene

13. Benzo(g,h,i)perylene 35. C1-naphthalenes

14. C1-fluorenes 36. C2-naphthalenes

15. C2-fluorenes 37. C3-naphthalenes

16. C3-fluorenes 38. C4-naphthalenes 

17. C1-phenanthrenes 39. C1-dibenzothiophenes

18. C2-phenanthrenes 40. C3-dibenzothiophenes

19. C3-phenanthrenes 41. C1-chrysenes 

20. C4-phenanthrenes 42. C2-chrysenes

21. C1-fluoranthenes+C1-pyrene 43. C3-chrysenes

22. C2-dibenzothiophenes 44. C4-chrysenes

Organochlorines

1. Lindane 15. HCB

2. Heptachlor 16. alpha BHC

3. Aldrin 17. beta BHC

4. Heptachlor epoxide 18. delta BHC

5. Chlordane 19. Oxychlordane

6. Endo 20. gamma Chlordane

7. Dieldrin 21. trans-nonachlor

8. DDE 22. PCB 1242

9. Endrin 23. PCB 1248

10. Perthane 24. PCB 1254

11. DDD 25. PCB 1260

12. DDT 26. alpha Chlordane

13. Methoxychlor 27. o.p’ DDD

14. Mirex 28. cis-nonchlor

15. Toxaphene 29. o,p’ DDT

16. o.p’ DDE
1.35

Appendix 1.4. Amphipod length data for the 1st set of sediment samples. Replication (Rep), Animal (individual animal number),

and length (mean length for individual animal; n=2 measurements).

 

Sample Animal Rep Length Sample Animal Rep Length 

 

ARCH 1 NA 0.946 1C 2 A 3.610

ARCH 2 NA 1.077 1C 3 A 3.475

ARCH 3 NA 1.134 1C 4 A 3.783

ARCH 4 NA 1.092 1C 5 A 3.669

ARCH 5 NA 0.973 1C 1 B 2.794

ARCH 6 NA 1.024 1C 2 B 3.616

ARCH 7 NA 1.122 1C 3 B 3.102

ARCH 8 NA 1.086 1C 4 B 4.112

ARCH 9 NA 1.000 1C 5 B 3.078

ARCH 10 NA 1.051 1C 6 B 2.946

ARCH 11 NA 1.086 1C 1 C 2.157

1B 1 A 3.373 1C 2 C 2.656

1B 2 A 2.522 1C 3 C 2.943

1B 3 A 3.048 1C 4 C 2.271

1B 4 A 3.084 1C 5 C 3.445

1B 5 A 3.610 1C 6 C 2.695

1B 6 A 3.090 1C 7 C 2.531

1B 7 A 3.655 1C 1 D 2.725

1B 8 A 3.265 1C 2 D 3.433

1B 1 B 3.843 1C 3 D 4.076

1B 2 B 3.666 1C 4 D 3.616

1B 3 B 4.348 1C 5 D 3.454

1B 4 B 3.630 1C 6 D 2.656

1B 5 B 3.783 3B 1 A 4.595

1B 6 B 3.765 3B 2 A 4.456

1B 7 B 4.207 3B 3 A 4.441

1B 8 B 3.556 3B 4 A 3.690

1B 9 B 3.846 3B 1 B 5.497

1B 10 B 3.332 3B 2 B 4.119

1B 1 C 4.398 3B 3 B 4.885

1B 2 C 4.889 3B 4 B 4.985

1B 3 C 3.864 3B 5 B 4.388

1B 4 C 4.646 3B 6 B 4.077

1B 5 C 3.409 3B 7 B 4.607

1B 6 C 4.942 3B 8 B 4.030

1B 7 C 4.073 3B 1 C 4.296

1B 8 C 4.883 3B 2 C 4.118

1B 9 C 4.222 3B 3 C 4.335

1B 1 D 3.173 3B 4 C 4.036

1B 2 D 2.925 3B 5 C 4.935

1B 3 D 2.474 3B 6 C 4.068

1B 4 D 4.282 3B 7 C 4.027

1B 5 D 2.752 3B 8 C 3.879

1B 6 D 3.179 3B 9 C 3.891

1B 7 D 3.164 3B 10 C 3.891

1B 8 D 3.472 3B 11 C 3.923

1B 9 D 3.215 3B 1 D 4.089

1C 1 A 3.170 3B 2 D 4.476
Appendix 1.4. Amphipod length data for the 1st set of sediment samples (continued).

 

Sample Animal Rep Length Sample Animal Rep Length 

 

1.36

3B 3 D 4.053 5C 5 B 3.388

3B 4 D 4.181 5C 6 B 3.099

3B 5 D 3.805 5C 1 C 3.822

3B 6 D 3.778 5C 2 C 3.825

3B 7 D 4.867 5C 3 C 3.762

3B 8 D 3.920 5C 4 C 3.750

5B 1 A 3.974 5C 5 C 4.046

5B 2 A 3.825 5C 6 C 3.944

5B 3 A 4.037 5C 7 C 3.995

5B 4 A 4.103 5C 8 C 4.700

5B 5 A 4.754 5C 9 C 3.553

5B 6 A 4.249 5C 1 D 4.667

5B 7 A 4.357 5C 2 D 4.617

5B 8 A 3.669 5C 3 D 4.569

5B 9 A 4.112 5C 4 D 4.327

5B 1 B 4.467 5C 5 D 5.295

5B 2 B 3.965 5C 6 D 4.019

5B 3 B 4.198 5C 7 D 4.431

5B 4 B 3.834 5C 8 D 3.801

5B 5 B 4.524 5C 9 D 4.482

5B 6 B 4.404 8B 1 A 4.443

5B 7 B 4.216 8B 2 A 3.867

5B 8 B 4.070 8B 3 A 3.887

5B 9 B 3.971 8B 4 A 3.517

5B 1 C 4.682 8B 5 A 3.858

5B 2 C 4.088 8B 6 A 4.094

5B 3 C 4.387 8B 7 A 4.046

5B 4 C 4.987 8B 8 A 5.017

5B 5 C 4.216 8B 9 A 4.826

5B 6 C 5.265 8B 10 A 5.098

5B 7 C 3.732 8B 1 B 4.159

5B 1 D 4.422 8B 2 B 3.777

5B 2 D 4.088 8B 3 B 3.622

5B 3 D 4.159 8B 4 B 3.834

5B 4 D 4.261 8B 5 B 4.270

5B 5 D 4.091 8B 6 B 3.669

5B 6 D 4.162 8B 7 B 3.580

5C 1 A 3.251 8B 8 B 3.490

5C 2 A 4.216 8B 9 B 3.242

5C 3 A 4.073 8B 10 B 2.253

5C 4 A 3.230 8B 1 C 3.054

5C 5 A 4.512 8B 2 C 3.654

5C 6 A 5.157 8B 3 C 3.389

5C 7 A 3.765 8B 4 C 3.816

5C 1 B 4.192 8B 5 C 3.455

5C 2 B 3.696 8B 6 C 3.066

5C 3 B 3.974 8B 7 C 3.081

5C 4 B 3.672 8B 8 C 3.837
Appendix 1.4. Amphipod length data for the 1st set of sediment samples (continued).

 

Sample Animal Rep Length Sample Animal Rep Length 

 

1.37

8B 1 D 3.352 10B 5 A 4.694

8B 2 D 3.675 10B 6 A 4.288

8B 3 D 3.151 10B 7 A 3.702

8B 4 D 4.045 10B 8 A 4.733

8B 5 D 3.557 10B 9 A 4.403

8B 6 D 3.374 10B 10 A 3.995

8B 7 D 3.560 10B 1 B 4.440

8B 8 D 2.940 10B 2 B 3.738

8B 9 D 3.922 10B 3 B 3.741

8B 10 D 3.075 10B 4 B 4.415

8B 11 D 3.373 10B 5 B 4.781

8C 1 A 4.283 10B 6 B 3.675

8C 2 A 4.238 10B 7 B 4.161

8C 3 A 3.581 10B 8 B 4.252

8C 4 A 4.027 10B 9 B 5.477

8C 5 A 3.916 10B 1 C 4.025

8C 6 A 2.807 10B 2 C 4.724

8C 7 A 3.367 10B 3 C 3.922

8C 8 A 2.458 10B 4 C 3.665

8C 9 A 4.142 10B 5 C 3.687

8C 1 B 4.241 10B 6 C 4.636

8C 2 B 4.253 10B 7 C 3.696

8C 3 B 3.858 10B 1 D 4.412

8C 4 B 5.099 10B 2 D 6.042

8C 5 B 3.831 10B 3 D 4.155

8C 6 B 3.678 10B 4 D 3.892

8C 1 C 4.136 10B 5 D 4.781

8C 2 C 4.184 10B 6 D 4.512

8C 3 C 4.127 10B 7 D 3.959

8C 4 C 3.467 10B 8 D 4.276

8C 5 C 4.524 10B 9 D 4.739

8C 6 C 3.876 10B 10 D 4.001

8C 7 C 3.461 10C 1 A 3.641

8C 8 C 3.587 10C 2 A 3.829

8C 9 C 4.425 10C 3 A 4.573

8C 1 D 4.460 10C 4 A 4.052

8C 2 D 4.346 10C 5 A 3.989

8C 3 D 4.322 10C 6 A 3.396

8C 4 D 3.654 10C 1 B 3.321

8C 5 D 4.747 10C 2 B 3.944

8C 6 D 4.383 10C 3 B 4.086

8C 7 D 5.024 10C 4 B 3.411

8C 8 D 5.575 10C 5 B 3.647

8C 9 D 5.015 10C 6 B 3.844

10B 1 A 4.594 10C 1 C 3.826

10B 2 A 4.104 10C 2 C 3.575

10B 3 A 3.959 10C 3 C 4.122

10B 4 A 3.632 10C 4 C 4.815
Appendix 1.4. Amphipod length data for the 1st set of sediment samples (continued).

 

Sample Animal Rep Length Sample Animal Rep Length 

 

1.38

10C 5 C 4.691 11C 1 A 3.813

10C 6 C 3.947 11C 2 A 3.095

10C 7 C 3.620 11C 3 A 3.628

10C 8 C 3.638 11C 4 A 3.357

10C 9 C 4.270 11C 5 A 3.741

10C 1 D 3.807 11C 6 A 3.143

10C 2 D 4.255 11C 7 A 3.732

10C 3 D 3.650 11C 1 B 3.741

10C 4 D 3.811 11C 2 B 4.179

10C 5 D 2.858 11C 3 B 3.325

10C 6 D 3.623 11C 4 B 4.107

10C 7 D 4.180 11C 5 B 3.497

10C 8 D 3.547 11C 1 C 3.280

11B 1 A 4.589 11C 2 C 3.664

11B 2 A 4.051 11C 3 C 3.571

11B 3 A 4.333 11C 4 C 4.146

11B 4 A 4.164 11C 5 C 4.122

11B 5 A 4.152 11C 6 C 3.652

11B 6 A 4.262 11C 1 D 3.315

11B 7 A 4.066 11C 2 D 3.717

11B 8 A 3.950 11C 3 D 3.688

11B 1 B 3.828 11C 4 D 3.574

11B 2 B 4.220 11C 5 D 3.057

11B 3 B 5.000 12B 1 A 4.003

11B 4 B 3.768 12B 2 A 4.275

11B 5 B 4.119 12B 3 A 4.095

11B 6 B 4.244 12B 4 A 3.586

11B 7 B 3.408 12B 5 A 3.414

11B 8 B 3.661 12B 6 A 3.837

11B 9 B 4.235 12B 7 A 2.515

11B 1 C 4.357 12B 8 A 3.154

11B 2 C 4.057 12B 9 A 3.870

11B 3 C 4.878 12B 1 B 3.210

11B 4 C 4.351 12B 2 B 2.669

11B 5 C 5.122 12B 3 B 3.447

11B 6 C 4.408 12B 4 B 3.678

11B 7 C 4.351 12B 5 B 2.964

11B 8 C 5.006 12B 6 B 3.039

11B 9 C 5.116 12B 7 B 3.873

11B 1 D 3.479 12B 8 B 3.769

11B 2 D 4.116 12B 9 B 3.876

11B 3 D 4.661 12B 1 C 3.453

11B 4 D 4.432 12B 2 C 3.755

11B 5 D 4.577 12B 3 C 3.293

11B 6 D 4.360 12B 4 C 3.036

11B 7 D 5.027 12B 5 C 3.494

11B 8 D 4.137 12B 6 C 3.335

11B 9 D 4.217 12B 7 C 3.512
Appendix 1.4. Amphipod length data for the 1st set of sediment samples (continued).

 

Sample Animal Rep Length Sample Animal Rep Length 

 

1.39

12B 1 D 3.663 15B 5 A 3.293

12B 2 D 3.352 15B 6 A 3.801

12B 3 D 2.808 15B 7 A 3.341

12B 4 D 3.565 15B 8 A 3.968

12B 5 D 3.988 15B 9 A 4.216

12B 6 D 3.293 15B 1 B 3.873

12B 7 D 3.518 15B 2 B 4.736

12C 1 A 4.236 15B 3 B 3.813

12C 2 A 4.233 15B 4 B 3.699

12C 3 A 4.209 15B 5 B 3.995

12C 4 A 3.249 15B 6 B 4.401

12C 5 A 4.518 15B 7 B 4.533

12C 6 A 4.155 15B 8 B 4.073

12C 7 A 3.051 15B 1 C 3.364

12C 8 A 3.738 15B 2 C 3.811

12C 9 A 4.224 15B 3 C 3.485

12C 1 B 4.026 15B 4 C 4.006

12C 2 B 3.477 15B 5 C 4.003

12C 3 B 3.813 15B 6 C 2.624

12C 4 B 4.353 15B 7 C 3.926

12C 5 B 3.669 15B 8 C 4.036

12C 6 B 4.242 15B 9 C 3.953

12C 7 B 3.711 15B 1 D 3.281

12C 8 B 4.506 15B 2 D 3.355

12C 9 B 3.033 15B 3 D 3.444

12C 10 B 3.594 15B 4 D 3.447

12C 1 C 3.960 15B 5 D 3.267

12C 2 C 3.615 15B 6 D 3.314

12C 3 C 4.062 15B 7 D 3.550

12C 4 C 3.798 15B 8 D 3.494

12C 5 C 3.972 15C 1 A 3.169

12C 6 C 4.110 15C 2 A 2.908

12C 7 C 2.748 15C 3 A 3.033

12C 8 C 4.062 15C 4 A 4.287

12C 1 D 3.639 15C 1 B 4.240

12C 2 D 3.705 15C 2 B 3.278

12C 3 D 3.408 15C 3 B 3.497

12C 4 D 3.327 15C 4 B 3.391

12C 5 D 3.249 15C 5 B 3.796

12C 6 D 3.831 15C 6 B 4.219

12C 7 D 3.240 15C 7 B 4.459

12C 8 D 4.083 15C 1 C 4.932

12C 9 D 3.444 15C 2 C 4.764

12C 10 D 3.681 15C 3 C 3.698

15B 1 A 3.801 15C 4 C 3.352

15B 2 A 4.273 15C 5 C 3.538

15B 3 A 3.870 15C 6 C 3.355

15B 4 A 3.054 15C 7 C 3.113
Appendix 1.4. Amphipod length data for the 1st set of sediment samples (continued).

 

Sample Animal Rep Length Sample Animal Rep Length 

 

1.40

15C 8 C 4.077 16C 7 B 3.612

15C 1 D 3.234 16C 8 B 3.755

15C 2 D 3.589 16C 9 B 3.590

15C 3 D 3.059 16C 1 C 3.497

15C 4 D 3.391 16C 2 C 3.503

15C 5 D 3.793 16C 3 C 3.308

15C 6 D 3.166 16C 4 C 4.084

15C 7 D 3.722 16C 5 C 3.330

15C 8 D 3.056 16C 6 C 3.637

15C 9 D 3.204 16C 7 C 3.991

16B 1 A 3.196 16C 8 C 3.376

16B 2 A 3.507 16C 9 C 3.951

16B 3 A 3.547 16C 1 C 3.376

16B 4 A 4.435 16C 1 D 3.851

16B 5 A 3.302 16C 2 D 3.680

16B 6 A 3.485 16C 3 D 3.730

16B 7 A 3.581 16C 4 D 3.541

16B 8 A 3.245 16C 5 D 4.438

16B 9 A 2.920 16C 6 D 4.364

16B 1 B 3.097 16C 7 D 4.659

16B 2 B 3.838 16C 8 D 4.165

16B 3 B 3.941 16C 9 D 4.308

16B 4 B 4.510 16C 10 D 4.395

16B 5 B 3.072 16C 11 D 3.826

16B 1 C 4.078 21B 1 A 3.280

16B 2 C 3.889 21B 2 A 3.224

16B 3 C 3.805 21B 3 A 3.301

16B 4 C 3.625 21B 4 A 3.916

16B 5 C 4.134 21B 5 A 3.200

16B 6 C 3.917 21B 6 A 3.999

16B 1 D 4.280 21B 7 A 3.107

16B 2 D 4.230 21B 8 A 3.483

16B 3 D 3.699 21B 9 A 2.872

16B 4 D 3.733 21B 10 A 3.265

16B 5 D 3.764 21B 1 B 3.283

16B 6 D 3.864 21B 2 B 3.319

16C 1 A 4.261 21B 3 B 3.030

16C 2 A 3.494 21B 4 B 3.781

16C 3 A 4.469 21B 5 B 4.160

16C 4 A 3.901 21B 6 B 4.178

16C 5 A 3.929 21B 7 B 2.809

16C 6 A 2.770 21B 8 B 3.775

16C 1 B 4.581 21B 9 B 3.856

16C 2 B 3.327 21B 1 C 3.480

16C 3 B 3.777 21B 2 C 3.161

16C 4 B 3.907 21B 3 C 3.808

16C 5 B 3.578 21B 4 C 3.579

16C 6 B 4.009 21B 5 C 4.056
Appendix 1.4. Amphipod length data for the 1st set of sediment samples (continued).

 

Sample Animal Rep Length Sample Animal Rep Length 

 

1.41

21B 6 C 3.501 25B 1 A 2.579

21B 7 C 3.960 25B 2 A 4.374

21B 8 C 3.579 25B 3 A 3.433

21B 1 D 3.611 25B 4 A 3.347

21B 2 D 2.887 25B 5 A 3.735

21B 3 D 3.140 25B 6 A 2.955

21B 4 D 3.632 25B 1 B 3.777

21B 5 D 2.565 25B 2 B 3.726

21B 6 D 3.543 25B 3 B 3.840

21B 7 D 3.811 25B 4 B 3.735

21B 8 D 3.012 25B 5 B 4.225

21B 9 D 3.537 25B 6 B 4.237

21B 10 D 3.254 25B 7 B 4.129

21C 1 A 4.595 25B 1 C 3.816

21C 2 A 3.895 25B 2 C 2.854

21C 3 A 4.476 25B 3 C 3.675

21C 4 A 3.397 25B 4 C 4.094

21C 5 A 4.512 25B 5 C 3.923

21C 6 A 3.343 25B 1 D 3.837

21C 7 A 4.509 25B 2 D 2.561

21C 8 A 3.850 25B 3 D 3.565

21C 9 A 3.808 25B 4 D 3.430

21C 1 B 3.069 25B 5 D 2.976

21C 2 B 3.358 25C 1 A 2.952

21C 3 B 2.863 25C 2 A 3.120

21C 4 B 3.295 25C 3 A 3.953

21C 5 B 3.069 25C 4 A 3.831

21C 6 B 3.671 25C 5 A 4.052

21C 7 B 3.110 25C 6 A 3.834

21C 8 B 3.376 25C 7 A 3.729

21C 1 C 3.865 25C 8 A 3.093

21C 2 C 4.539 25C 1 B 5.053

21C 3 C 4.109 25C 2 B 3.920

21C 4 C 4.366 25C 1 C 3.547

21C 5 C 4.921 25C 2 C 3.487

21C 6 C 2.920 25C 3 C 3.469

21C 7 C 3.901 25C 4 C 3.299

21C 8 C 3.620 25C 5 C 3.215

21C 1 D 4.047 25C 6 C 3.681

21C 2 D 4.273 25C 7 C 3.356

21C 3 D 3.373 25C 8 C 3.108

21C 4 D 4.545 25C 9 C 3.571

21C 5 D 3.987 25C 10 C 3.448

21C 6 D 3.957 25C 1 D 3.645

21C 7 D 4.643 25C 2 D 4.494

21C 8 D 4.094 25C 3 D 3.344

21C 9 D 4.050 25C 4 D 3.538

21C 10 D 4.088 25C 5 D 3.729
Appendix 1.4. Amphipod length data for the 1st set of sediment samples (continued).

 

Sample Animal Rep Length Sample Animal Rep Length 

 

1.42

25C 6 D 3.266 26C 10 A 2.829

25C 7 D 3.648 26C 1 B 2.764

25C 8 D 3.887 26C 2 B 3.267

25C 9 D 3.953 26C 3 B 3.084

26B 1 A 3.090 26C 4 B 2.713

26B 2 A 3.837 26C 5 B 2.875

26B 3 A 3.750 26C 6 B 3.171

26B 4 A 3.642 26C 7 B 3.006

26B 5 A 4.744 26C 1 C 2.761

26B 6 A 4.735 26C 2 C 2.686

26B 7 A 4.163 26C 3 C 2.731

26B 8 A 3.319 26C 4 C 2.883

26B 9 A 4.072 26C 5 C 2.632

26B 10 A 4.603 26C 6 C 2.680

26B 1 B 3.883 26C 1 D 3.204

26B 2 B 4.253 26C 2 D 3.012

26B 3 B 2.313 26C 3 D 2.665

26B 4 B 3.762 26C 4 D 2.620

26B 5 B 3.139 26C 5 D 2.593

26B 6 B 3.536 26C 6 D 2.764

26B 7 B 3.825 26C 7 D 2.964

26B 8 B 4.184 26C 8 D 3.054

26B 1 C 3.072 26C 9 D 2.958

26B 2 C 2.958 26C 10 D 2.255

26B 3 C 3.293 FLOR 1 A 2.734

26B 4 C 2.922 FLOR 2 A 4.479

26B 5 C 3.338 FLOR 3 A 3.796

26B 6 C 3.039 FLOR 4 A 3.362

26B 7 C 3.533 FLOR 5 A 4.323

26B 8 C 2.961 FLOR 6 A 3.401

26B 9 C 3.003 FLOR 7 A 3.826

26B 1 D 2.946 FLOR 1 B 4.156

26B 2 D 3.740 FLOR 2 B 3.087

26B 3 D 3.434 FLOR 3 B 3.237

26B 4 D 3.219 FLOR 4 B 3.278

26B 5 D 3.084 FLOR 5 B 4.587

26B 6 D 2.931 FLOR 6 B 2.455

26B 7 D 2.925 FLOR 7 B 3.332

26B 8 D 3.344 FLOR 8 B 3.434

26C 1 A 2.701 FLOR 1 C 2.731

26C 2 A 3.018 FLOR 2 C 1.338

26C 3 A 3.039 FLOR 3 C 1.976

26C 4 A 2.832 FLOR 4 C 3.683

26C 5 A 2.659 FLOR 5 C 3.668

26C 6 A 3.341 FLOR 6 C 2.973

26C 7 A 3.063 FLOR 7 C 1.868

26C 8 A 3.012 FLOR 1 D 4.341

26C 9 A 3.126 FLOR 2 D 3.790
Appendix 1.4. Amphipod length data for the 1st set of sediment samples (continued).

 

Sample Animal Rep Length Sample Animal Rep Length 

 

1.43

FLOR 3 D 3.644 FLOR 7 D 3.976

FLOR 4 D 4.072 FLOR 8 D 4.231

FLOR 5 D 2.578 FLOR 9 D 1.967

FLOR 6 D 4.760

 
1.44

Appendix 1.5. Amphipod length data for the 2nd set of samples. Replication (Rep), Animal (individual animal number), and length

(mean length for individual animal).

 

Sample Animal Rep Length Sample Animal Rep Length 

 

ARCH 1 NA 1.339 2C 6 B 3.066

ARCH 2 NA 1.369 2C 7 B 3.475

ARCH 3 NA 1.193 2C 8 B 3.200

ARCH 4 NA 1.178 2C 9 B 2.755

ARCH 5 NA 1.101 2C 10 B 3.956

ARCH 6 NA 1.107 2C 1 C 2.848

ARCH 7 NA 1.021 2C 2 C 3.938

ARCH 8 NA 1.056 2C 3 C 4.109

ARCH 9 NA 1.134 2C 4 C 3.884

ARCH 10 NA 1.196 2C 5 C 3.514

2B 1 A 4.177 2C 6 C 3.804

2B 2 A 3.556 2C 7 C 4.046

2B 3 A 3.890 2C 1 D 4.593

2B 4 A 4.572 2C 2 D 3.093

2B 5 A 4.001 2C 3 D 2.352

2B 6 A 3.920 2C 4 D 4.001

2B 7 A 4.467 2C 5 D 3.968

2B 8 A 4.195 2C 6 D 4.443

2B 1 B 4.461 2C 7 D 3.117

2B 2 B 4.491 2C 8 D 4.234

2B 3 B 3.729 2C 9 D 4.243

2B 4 B 4.246 4B 1 A 3.616

2B 5 B 4.622 4B 2 A 4.485

2B 6 B 3.502 4B 3 A 4.467

2B 7 B 4.718 4B 4 A 3.174

2B 8 B 3.421 4B 1 B 3.337

2B 1 C 5.184 4B 2 B 3.243

2B 2 C 3.980 4B 3 B 3.295

2B 3 C 5.558 4B 4 B 2.579

2B 4 C 4.213 4B 5 B 3.391

2B 1 D 2.874 4B 6 B 3.343

2B 2 D 4.548 4B 7 B 4.148

2B 3 D 3.027 4B 8 B 2.841

2B 4 D 3.711 4B 1 C 3.415

2B 5 D 3.568 4B 2 C 3.457

2B 6 D 4.040 4B 3 C 3.270

2B 7 D 3.777 4B 4 C 3.682

2B 8 D 3.436 4B 5 C 3.944

2C 1 A 3.108 4B 6 C 2.291

2C 2 A 2.898 4B 7 C 3.433

2C 3 A 2.737 4B 8 C 3.388

2C 4 A 2.880 4B 9 C 4.413

2C 5 A 3.102 4B 1 D 2.600

2C 1 B 3.096 4B 2 D 2.444

2C 2 B 3.442 4B 3 D 3.860

2C 3 B 3.359 4B 4 D 4.205

2C 4 B 3.403 4B 5 D 3.240

2C 5 B 2.764 4B 6 D 2.850
Appendix 1.5. Amphipod length data for the 2nd set of sediment samples (continued).

 

Sample Animal Rep Length Sample Animal Rep Length 

 

1.45

4B 7 D 3.502 6B 6 C 4.360

4B 8 D 3.213 6B 7 C 3.045

4B 9 D 2.714 6B 8 C 3.200

4B 10 D 3.821 6B 9 C 3.242

4B 11 D 3.478 6B 10 C 3.831

4B 12 D 2.868 6B 1 D 2.943

4B 13 D 2.922 6B 2 D 3.311

4B 14 D 3.785 6B 3 D 3.935

4C 1 B 3.024 6B 4 D 4.634

4C 2 B 2.744 6B 5 D 3.553

4C 3 B 2.594 6B 6 D 2.994

4C 4 B 3.661 6B 7 D 3.335

4C 5 B 2.802 6B 8 D 3.777

4C 6 B 3.195 6C 1 A 3.568

4C 7 B 2.934 6C 2 A 3.819

4C 8 B 3.556 6C 3 A 2.943

4C 9 B 3.562 6C 4 A 4.204

4C 10 B 3.234 6C 5 A 4.336

4C 11 B 3.439 6C 6 A 5.462

4C 1 C 3.093 6C 7 A 3.027

4C 2 C 3.030 6C 8 A 4.608

4C 3 C 4.269 6C 9 A 4.019

4C 4 C 3.036 6C 1 B 4.790

4C 5 C 3.039 6C 2 B 4.775

4C 6 C 3.986 6C 3 B 4.183

4C 7 C 3.830 6C 4 B 3.870

4C 8 C 3.144 6C 5 B 4.688

4C 1 D 3.493 6C 6 B 4.760

4C 2 D 3.758 6C 7 B 4.617

4C 3 D 4.323 6C 8 B 3.855

4C 4 D 3.721 6C 1 C 3.583

4C 5 D 3.427 6C 2 C 4.682

4C 6 D 3.195 6C 3 C 3.439

4C 7 D 3.090 6C 4 C 3.317

6B 1 A 3.317 6C 5 C 4.969

6B 2 A 3.508 6C 6 C 3.601

6B 3 A 3.499 6C 7 C 3.589

6B 4 A 3.326 6C 8 C 4.115

6B 5 A 3.132 6C 1 D 3.765

6B 6 A 3.493 6C 2 D 3.911

6B 7 A 2.949 6C 3 D 3.622

6B 1 B 3.024 6C 4 D 3.941

6B 2 B 3.908 6C 5 D 4.318

6B 1 C 3.132 6C 6 D 4.464

6B 2 C 3.636 6C 7 D 4.085

6B 3 C 4.013 6C 8 D 3.657

6B 4 C 4.443 7B 1 A 3.886

6B 5 C 3.741 7B 2 A 3.844
Appendix 1.5. Amphipod length data for the 2nd set of sediment samples (continued).

 

Sample Animal Rep Length Sample Animal Rep Length 

 

1.46

7B 3 A 3.605 7C 2 B 3.593

7B 4 A 3.725 7C 3 B 3.889

7B 5 A 3.635 7C 4 B 3.859

7B 6 A 3.695 7C 5 B 3.546

7B 7 A 3.361 7C 6 B 4.172

7B 8 A 4.294 7C 7 B 4.232

7B 9 A 3.537 7C 8 B 3.304

7B 10 A 4.127 7C 9 B 3.895

7B 1 B 3.263 7C 10 B 4.348

7B 2 B 3.185 7C 1 C 3.507

7B 3 B 3.835 7C 2 C 3.450

7B 4 B 3.447 7C 3 C 4.578

7B 5 B 3.263 7C 4 C 3.725

7B 6 B 3.587 7C 5 C 3.701

7B 7 B 4.262 7C 6 C 3.435

7B 8 B 3.832 7C 7 C 3.656

7B 9 B 3.158 7C 8 C 3.534

7B 10 B 3.641 7C 9 C 3.898

7B 1 C 3.549 7C 1 D 3.602

7B 2 C 3.531 7C 2 D 3.743

7B 3 C 3.531 7C 3 D 4.005

7B 4 C 3.140 7C 4 D 3.925

7B 5 C 3.087 7C 5 D 3.811

7B 6 C 3.084 7C 6 D 3.671

7B 7 C 3.486 7C 7 D 3.352

7B 8 C 3.110 7C 8 D 3.307

7B 9 C 4.411 7C 9 D 3.477

7B 10 C 3.570 7C 10 D 3.632

7B 1 D 4.470 7C 11 D 1.813

7B 2 D 4.136 9B 1 A 4.027

7B 3 D 4.238 9B 2 A 4.675

7B 4 D 4.050 9B 3 A 4.033

7B 5 D 3.543 9B 4 A 3.801

7B 6 D 3.707 9B 5 A 3.178

7B 7 D 3.638 9B 6 A 3.587

7B 8 D 3.948 9B 7 A 3.443

7B 9 D 3.543 9B 1 B 3.301

7B 10 D 3.590 9B 2 B 3.398

7C 1 A 4.405 9B 3 B 5.358

7C 2 A 4.160 9B 4 B 4.527

7C 3 A 3.283 9B 5 B 3.346

7C 4 A 3.659 9B 1 C 3.214

7C 5 A 3.468 9B 2 C 3.455

7C 6 A 3.090 9B 3 C 3.705

7C 7 A 4.005 9B 4 C 3.844

7C 8 A 3.361 9B 5 C 3.870

7C 9 A 3.987 9B 6 C 3.654

7C 1 B 4.178 9B 7 C 3.843
Appendix 1.5. Amphipod length data for the 2nd set of sediment samples (continued).

 

Sample Animal Rep Length Sample Animal Rep Length 

 

1.47

9B 8 C 3.772 13B 1 C 4.581

9B 9 C 3.352 13B 2 C 3.388

9B 10 C 3.566 13B 3 C 4.079

9B 11 C 3.461 13B 4 C 3.308

9B 1 D 3.662 13B 5 D 3.006

9B 2 D 3.759 13B 1 D 2.766

9B 3 D 2.822 13B 2 D 3.970

9B 4 D 3.304 13B 3 D 3.635

9B 5 D 4.416 13B 4 D 3.701

9B 6 D 3.753 13C 1 A 3.968

9B 7 D 3.699 13C 2 A 3.553

9B 8 D 3.361 13C 3 A 3.460

9C 1 A 2.913 13C 1 B 3.364

9C 2 A 2.976 13C 2 B 3.290

9C 3 A 3.666 13C 3 B 3.905

9C 4 A 4.283 13C 4 B 4.138

9C 1 B 4.093 13C 5 B 3.759

9C 2 B 3.843 13C 6 B 3.977

9C 3 B 3.374 13C 1 C 2.940

9C 4 B 4.202 13C 2 C 3.448

9C 5 B 4.081 13C 3 C 2.851

9C 6 B 3.708 13C 4 C 4.643

9C 7 B 4.280 13C 5 C 3.565

9C 8 B 4.018 13C 6 C 2.737

9C 1 C 3.072 13C 1 D 3.478

9C 2 C 3.229 13C 2 D 3.565

9C 3 C 3.334 13C 3 D 3.451

9C 4 C 3.982 14B 1 A 3.092

9C 5 C 3.681 14B 2 A 3.693

9C 6 C 3.509 14B 3 A 3.503

9C 7 C 3.178 14B 4 A 3.235

9C 8 C 3.340 14B 5 A 4.714

9C 9 C 3.566 14B 6 A 3.271

9C 10 C 3.692 14B 7 A 3.024

9C 11 C 3.162 14B 1 B 4.211

9C 12 C 2.825 14B 2 B 3.429

9C 13 C 3.289 14B 3 B 4.274

9C 14 C 3.656 14B 4 B 4.482

9C 15 C 4.075 14B 5 B 3.786

9C 16 C 4.256 14B 6 B 4.092

9C 1 D 4.100 14B 7 B 5.723

9C 2 D 3.991 14B 1 C 3.542

9C 3 D 3.256 14B 2 C 3.408

9C 4 D 4.142 14B 3 C 3.780

13B 1 A 5.304 14B 4 C 2.872

13B 2 A 4.062 14B 5 C 4.452

13B 1 B 4.652 14B 6 C 4.036

13B 2 B 3.870 14B 1 D 3.934
Appendix 1.5. Amphipod length data for the 2nd set of sediment samples (continued).

 

Sample Animal Rep Length Sample Animal Rep Length 

 

1.48

14B 2 D 3.173 18B 3 D 3.534

14B 3 D 3.756 18B 4 D 3.618

14B 4 D 3.863 18B 5 D 3.693

14B 5 D 4.821 18C 1 A 3.608

14B 6 D 4.036 18C 2 A 3.310

14B 7 D 3.863 18C 3 A 3.841

14C 1 A 3.434 18C 4 A 2.696

14C 2 A 3.066 18C 5 A 3.412

14C 3 A 4.009 18C 6 A 4.169

14C 4 A 3.268 18C 7 A 3.793

14C 5 A 4.053 18C 8 A 3.552

14C 6 A 3.426 18C 9 A 2.767

14C 1 B 2.568 18C 1 B 2.857

14C 2 B 3.009 18C 2 B 3.823

14C 3 B 2.640 18C 3 B 4.601

14C 4 B 2.631 18C 4 B 3.626

14C 5 B 3.765 18C 5 B 3.304

14C 6 B 4.054 18C 1 C 3.444

14C 1 C 3.527 18C 2 C 3.578

14C 2 C 4.161 18C 3 C 4.178

14C 3 C 3.134 18C 4 C 2.994

14C 4 C 3.946 18C 5 C 3.414

14C 1 D 3.934 18C 6 C 3.868

14C 2 D 3.845 18C 7 C 2.896

14C 3 D 4.077 18C 8 C 3.728

18B 1 A 3.816 18C 9 C 3.155

18B 2 A 3.375 18C 10 C 3.364

18B 3 A 2.355 18C 1 D 3.904

18B 4 A 4.167 19B 1 A 4.054

18B 5 A 3.978 19B 2 A 3.348

18B 6 A 3.000 19B 3 A 3.872

18B 7 A 2.559 19B 4 A 3.339

18B 1 B 4.803 19B 5 A 3.304

18B 2 B 3.636 19B 6 A 3.167

18B 3 B 2.898 19B 7 A 3.146

18B 4 B 3.681 19B 1 B 3.348

18B 5 B 3.738 19B 2 B 4.214

18B 6 B 4.503 19B 3 B 2.450

18B 7 B 4.020 19B 4 B 3.455

18B 1 C 3.618 19B 5 B 2.938

18B 2 C 2.664 19B 6 B 3.923

18B 3 C 3.711 19B 7 B 3.506

18B 4 C 3.579 19B 8 B 3.726

18B 5 C 3.570 19B 9 B 3.173

18B 6 C 2.946 19B 1 C 3.774

18B 7 C 2.928 19B 2 C 3.851

18B 1 D 4.065 19B 3 C 3.378

18B 2 D 4.434 19B 4 C 3.295
Appendix 1.5. Amphipod length data for the 2nd set of sediment samples (continued).

 

Sample Animal Rep Length Sample Animal Rep Length 

 

1.49

19B 5 C 2.818 20B 7 A 3.454

19B 6 C 3.711 20B 8 A 2.940

19B 7 C 2.970 20B 1 B 3.433

19B 8 C 3.205 20B 2 B 3.663

19B 1 D 3.009 20B 3 B 3.577

19B 2 D 3.304 20B 4 B 3.837

19B 3 D 3.319 20B 5 B 3.269

19B 4 D 3.015 20B 6 B 3.317

19B 5 D 2.774 20B 7 B 3.768

19B 6 D 3.089 20B 8 B 3.281

19B 7 D 3.545 20B 9 B 4.132

19B 8 D 2.616 20B 10 B 3.995

19B 9 D 2.836 20B 1 C 3.122

19B 10 D 2.917 20B 2 C 2.510

19C 1 A 3.893 20B 3 C 2.854

19C 2 A 3.792 20B 4 C 4.183

19C 3 A 3.238 20B 5 C 3.155

19C 4 A 2.929 20B 6 C 3.442

19C 5 A 3.581 20B 7 C 3.380

19C 6 A 2.515 20B 8 C 3.353

19C 7 A 3.512 20B 1 D 2.582

19C 8 A 4.137 20B 2 D 2.872

19C 1 B 3.631 20B 3 D 2.883

19C 2 B 3.777 20B 4 D 3.433

19C 3 B 2.997 20B 5 D 3.738

19C 4 B 3.176 20B 6 D 3.030

19C 5 B 3.875 20C 1 A 3.669

19C 6 B 2.979 20C 2 A 3.807

19C 1 C 3.646 20C 3 A 3.454

19C 2 C 3.256 20C 4 A 4.138

19C 3 C 3.435 20C 5 A 3.403

19C 4 C 3.955 20C 6 A 3.750

19C 5 C 3.438 20C 7 A 3.634

19C 6 C 3.307 20C 8 A 3.030

19C 7 C 3.149 20C 9 A 3.466

19C 8 C 3.720 20C 1 B 3.370

19C 1 D 2.920 20C 2 B 3.379

19C 2 D 3.845 20C 3 B 2.794

19C 3 D 3.366 20C 4 B 3.466

19C 4 D 3.494 20C 5 B 3.364

19C 5 D 3.173 20C 6 B 3.547

19C 6 D 3.485 20C 7 B 2.749

20B 1 A 3.630 20C 8 B 3.484

20B 2 A 3.580 20C 9 B 3.457

20B 3 A 4.004 20C 1 C 3.472

20B 4 A 3.711 20C 2 C 2.994

20B 5 A 3.834 20C 3 C 3.374

20B 6 A 3.884 20C 4 C 3.320
Appendix 1.5. Amphipod length data for the 2nd set of sediment samples (continued).

 

Sample Animal Rep Length Sample Animal Rep Length 

 

1.50

20C 5 C 3.350 22B 6 D 3.873

20C 6 C 3.418 22C 1 A 3.914

20C 7 C 3.143 22C 2 A 4.025

20C 8 C 3.021 22C 3 A 4.679

20C 9 C 3.780 22C 1 B 3.335

20C 1 D 2.979 22C 2 B 3.813

20C 2 D 2.964 22C 3 B 3.126

20C 3 D 3.230 22C 4 B 3.063

20C 4 D 3.185 22C 5 B 3.598

20C 5 D 3.388 22C 6 B 3.601

20C 6 D 2.522 22C 7 B 2.674

20C 7 D 3.182 22C 8 B 3.935

20C 8 D 3.015 22C 1 C 3.457

20C 9 D 3.221 22C 2 C 3.998

20C 10 D 2.564 22C 3 C 3.427

22B 1 A 3.586 22C 4 C 2.755

22B 2 A 3.708 22C 5 C 3.391

22B 3 A 3.786 22C 1 D 3.610

22B 4 A 4.004 22C 2 D 3.227

22B 5 A 3.063 22C 3 D 4.144

22B 6 A 3.550 22C 4 D 3.436

22B 7 A 3.350 22C 5 D 4.856

22B 8 A 4.121 22C 6 D 3.311

22B 9 A 3.415 22C 7 D 4.252

22B 10 A 4.778 24B 1 A 3.925

22B 1 B 5.253 24B 2 A 2.958

22B 2 B 4.351 24B 3 A 3.886

22B 3 B 3.666 24B 4 A 3.617

22B 4 B 3.732 24B 5 A 3.533

22B 5 B 3.281 24B 6 A 3.781

22B 6 B 3.173 24B 7 A 3.434

22B 7 B 3.885 24B 8 A 4.302

22B 8 B 3.290 24B 1 B 2.263

22B 9 B 2.848 24B 2 B 3.165

22B 1 C 3.188 24B 3 B 3.820

22B 2 C 3.639 24B 4 B 4.159

22B 3 C 4.760 24B 5 B 4.108

22B 4 C 3.726 24B 6 B 3.536

22B 5 C 4.527 24B 7 B 4.054

22B 6 C 4.258 24B 8 B 3.599

22B 7 C 4.978 24B 9 B 3.329

22B 8 C 2.934 24B 1 C 3.293

22B 9 C 3.729 24B 2 C 4.045

22B 1 D 3.595 24B 3 C 4.362

22B 2 D 3.753 24B 4 C 2.931

22B 3 D 3.556 24B 5 C 3.012

22B 4 D 3.917 24B 6 C 3.707

22B 5 D 3.636 24B 7 C 3.922
Appendix 1.5. Amphipod length data for the 2nd set of sediment samples (continued).

 

Sample Animal Rep Length Sample Animal Rep Length 

 

1.51

24B 8 C 3.189 SCB 5 C 2.784

24B 9 C 3.257 SCB 6 C 2.651

24B 1 D 3.862 SCB 7 C 3.661

24B 2 D 3.877 SCB 1 D 3.605

24B 3 D 3.563 SCB 2 D 3.708

24B 4 D 3.931 SCB 3 D 4.795

24B 5 D 3.054 SCB 4 D 3.343

24B 6 D 3.060 SCB 5 D 3.352

24B 7 D 3.653 SCB 6 D 2.955

24B 8 D 3.722 SCC 1 A 3.373

24B 9 D 4.311 SCC 2 A 2.286

24C 1 A 3.877 SCC 3 A 2.506

24C 2 A 3.750 SCC 4 A 2.973

24C 3 A 4.007 SCC 5 A 2.678

24C 4 A 3.490 SCC 6 A 3.051

24C 5 A 2.985 SCC 7 A 3.331

24C 6 A 3.414 SCC 8 A 3.238

24C 7 A 3.481 SCC 1 B 3.259

24C 8 A 3.980 SCC 2 B 2.955

24C 1 B 2.822 SCC 3 B 3.346

24C 2 B 3.820 SCC 4 B 3.563

24C 3 B 3.841 SCC 5 B 3.518

24C 4 B 4.542 SCC 6 B 2.952

24C 5 B 3.611 SCC 7 B 3.111

24C 6 B 4.836 SCC 8 B 2.961

24C 1 C 3.157 SCC 1 C 2.789

24C 2 C 3.859 SCC 2 C 2.584

24C 3 C 3.505 SCC 3 C 2.732

24C 4 C 3.832 SCC 4 C 2.753

24C 5 C 3.756 SCC 5 C 2.937

24C 6 C 3.139 SCC 6 C 2.861

24C 1 D 4.001 SCC 7 C 2.855

24C 2 D 5.571 SCC 8 C 2.922

24C 3 D 3.387 SCC 9 C 3.352

24C 4 D 4.083 SCC 10 C 2.404

SCB 1 A 3.794 SCC 1 D 2.725

SCB 2 A 3.490 SCC 2 D 3.275

SCB 3 A 3.758 SCC 3 D 3.359

SCB 4 A 3.171 SCC 4 D 3.741

SCB 1 B 3.748 SCC 5 D 3.648

SCB 2 B 3.736 SCC 6 D 2.537

SCB 3 B 2.994 SCC 7 D 2.523

SCB 4 B 3.239 SCC 8 D 3.143

SCB 5 B 3.393 SCC 9 D 3.648

SCB 1 C 3.325 FLOR1 A 2.334

SCB 2 C 2.933 FLOR2 A 2.522

SCB 3 C 3.319 FLOR3 A 2.256

SCB 4 C 3.583 FLOR4 A 2.561
1.52

FLOR 5 A 2.624 FLOR 4 C 3.606

FLOR 6 A 2.546 FLOR 5 C 2.444

FLOR 7 A 1.743 FLOR 6 C 2.916

FLOR 8 A 1.960 FLOR 7 C 2.866

FLOR 9 A 4.070 FLOR 8 C 2.761

FLOR 1 B 2.394 FLOR 1 D 2.113

FLOR 2 B 1.978 FLOR 2 D 1.909

FLOR 3 B 2.913 FLOR 3 D 2.576

FLOR 4 B 2.949 FLOR 4 D 1.790

FLOR 5 B 2.650 FLOR 5 D 2.283

FLOR 6 B 2.850 FLOR 6 D 2.360

FLOR 7 B 2.901 FLOR 7 D 2.522

FLOR 8 B 3.436 FLOR 8 D 3.347

FLOR 9 B 2.268 FLOR 9 D 2.668

FLOR 10 B 2.531 FLOR 10 D 2.059

FLOR 1 C 3.320 FLOR 11 D 1.981

FLOR 2 C 2.328

FLOR 3 C 3.230

 
1.53

Appendix 1.6. Amphipod maturation and survival data for the 1st set of samples. Replication (Rep), number of amphipods

recovered (Recov), and number of males recovered (Males).

 

Sample Rep Recov Males Sample Rep Recov Males 

 

01B 1 8 3 12C 1 9 4

01B 2 10 3 12C 2 10 2

01B 3 9 5 12C 3 8 2

01B 4 9 3 12C 4 10 4

01C 1 5 1 15B 1 9 3

01C 2 6 0 15B 2 8 6

01C 3 7 1 15B 3 9 6

01C 4 6 2 15B 4 10 3

03B 1 4 1 15C 1 7 1

03B 2 8 3 15C 2 7 2

03B 3 11 6 15C 3 8 3

03B 4 8 5 15C 4 9 5

05B 1 9 2 16B 1 9 2

05B 2 9 6 16B 2 5 2

05B 3 7 4 16B 3 6 3

05B 4 6 2 16B 4 6 3

05C 1 7 1 16C 1 6 1

05C 2 6 1 16C 2 9 1

05C 3 9 3 16C 3 9 5

05C 4 9 2 16C 4 11 4

08B 1 10 5 21B 1 10 5

08B 2 10 2 21B 2 9 4

08B 3 8 3 21B 3 9 4

08B 4 11 6 21B 4 10 7

08C 1 9 3 21C 1 9 5

08C 2 6 3 21C 2 8 3

08C 3 8 1 21C 3 8 5

08C 4 9 3 21C 4 10 5

10B 1 10 5 25B 1 6 1

10B 2 9 2 25B 2 7 2

10B 3 7 6 25B 3 5 0

10B 4 10 0 25B 4 4 2

10C 1 6 3 25C 1 8 2

10C 2 6 1 25C 2 2 1

10C 3 9 3 25C 3 10 1

10C 4 8 3 25C 4 9 3

11B 1 7 2 26B 1 10 5

11B 2 9 3 26B 2 8 2

11B 3 9 3 26B 3 9 5

11B 4 9 7 26B 4 8 3

11C 1 7 1 26C 1 10 7

11C 2 5 4 26C 2 7 2

11C 3 6 1 26C 3 6 4

11C 4 5 1 26C 4 10 3

12B 1 9 3 FLB 1 6 2

12B 2 9 3 FLB 2 8 5

12B 3 7 2 FLB 3 7 2

12B 4 7 3 FLB 4 9 2 
1.54

Appendix 1.7. Amphipod maturation and survival data for the 2nd set of samples. Replication (Rep), number of amphipods

recovered (Recov), and number of males recovered (Males).

 

Sample Rep Recov Males Sample Rep Recov Males 

 

02B 1 8 4 14B 1 7 1

02B 2 8 2 14B 2 7 2

02B 3 4 1 14B 3 6 3

02B 4 8 2 14B 4 6 2

02C 1 5 2 14C 1 6 1

02C 2 10 4 14C 2 6 1

02C 3 7 2 14C 3 4 3

02C 4 9 6 14C 4 3 2

04B 1 4 3 18B 1 7 2

04B 2 8 2 18B 2 7 4

04B 3 9 1 18B 3 7 1

04B 4 14 5 18B 4 5 5

04C 1 0 0 18C 1 9 3

04C 2 11 1 18C 2 5 1

04C 3 8 2 18C 3 10 3

04C 4 7 1 18C 4 1 0

06B 1 7 1 19B 1 7 3

06B 2 2 1 19B 2 9 7

06B 3 10 1 19B 3 8 0

06B 4 6 2 19B 4 10 4

06C 1 9 5 19C 1 8 3

06C 2 8 4 19C 2 6 3

06C 3 8 4 19C 3 8 2

06C 4 8 5 19C 4 6 1

07B 1 10 2 20B 1 8 2

07B 2 10 7 20B 2 10 1

07B 3 10 4 20B 3 8 1

07B 4 10 4 20B 4 6 0

07C 1 9 3 20C 1 9 4

07C 2 10 3 20C 2 9 0

07C 3 9 3 20C 3 9 4

07C 4 11 5 20C 4 10 2

09B 1 7 4 22B 1 10 2

09B 2 5 2 22B 2 9 2

09B 3 11 3 22B 3 9 2

09B 4 8 4 22B 4 6 2

09C 1 4 2 22C 1 3 2

09C 2 8 3 22C 2 8 4

09C 3 16 7 22C 3 5 0

09C 4 4 0 22C 4 7 3

13B 1 2 0 24B 1 8 3

13B 2 2 1 24B 2 9 2

13B 3 5 0 24B 3 9 4

13B 4 4 1 24B 4 9 3

13C 1 3 2 24C 1 8 3

13C 2 6 2 24C 2 5 4

13C 3 6 2 24C 3 6 6

13C 4 3 2 24C 4 4 2 
1.55

Appendix 1.7. (Continued)

 

Sample Rep Recov Males Sample Rep Recov Males 

 

SCB 1 4 0 FLC 1 9 1

SCB 2 5 0 FLC 2 10 0

SCB 3 7 1 FLC 3 8 1

SCB 4 6 2 FLC 4 11 0

SCC 1 8 2

SCC 2 8 3

SCC 3 10 2

SCC 4 9 4 
1.56

Appendix 1.8. Concentrations of acid volatile sulfide (μmoles/g) and simultaneously extracted metals (SEM in ug/g dry weight) and the sum of 

and the sum of the molar concentration of SEM divided by the molar concentration of AVS for the Upper Mississippi River sediment samples.

Sample AVS Cd Cu Ni Pb Zn 3SEM 3SEM/A

1B 0.19 0.058 1.1 1.2 5.2 11 0.232 1.221C 0.005 2.1 0.67 0.76 4.3 7 0.170 17.002B 1.9 0.39 4.7 4.6 7.7 26 0.591 0.312C 3.1 0.91 8.4 7.6 13.0 47 1.051 0.343B 2.1 0.68 7.3 6.1 11.0 35 0.813 0.394B 9.4 1.5 13.0 9.9 26.0 73 1.629 0.174C 16.0 3.0 20.0 15.0 53.0 118 2.658 0.175B 1.9 0.21 3.4 3.0 4.8 15 0.359 0.195C 16.0 0.63 6.3 9.4 16.0 45 1.030 0.066B 0.8 0.062 0.85 1.5 2.1 7.1 0.158 0.206C 2.9 0.12 2.0 2.7 4.2 12 0.282 0.107B 1.4 0.2 2.7 4.6 4.6 15 0.374 0.277C 8.7 0.43 5.3 6.6 12.0 35 0.793 0.098B 0.6 0.14 2.4 2.6 4.1 12 0.287 0.488C 2.2 0.28 4.1 4.6 6.9 20 0.485 0.229B 3.8 0.3 4.4 4.9 8.4 24 0.563 0.159C 3.8 0.38 5.2 5.4 11.0 31 0.705 0.1910B 4.8 0.17 3.7 3.2 6.0 18 0.419 0.0910C 63.0 0.74 11.0 10.0 26.0 68 1.516 0.0211B 1.1 0.13 2.5 3.2 5.8 16 0.368 0.3311C 6.5 0.18 2.8 3.8 7.1 17 0.405 0.0612B 3.8 0.43 4.8 4.8 26.0 137 2.382 0.6312C 5.9 0.45 4.5 4.1 27.0 143 2.462 0.4213B 3.1 0.14 2.3 3.2 6.8 22 0.461 0.1513C 5.2 0.3 4.4 4.5 14.0 48 0.950 0.1814B 3.5 0.14 2.8 3.3 6.1 19 0.422 0.1214C 18.0 0.41 6.8 6.6 18.0 51 1.090 0.0615B 3.6 0.2 4.0 3.9 9.9 31 0.653 0.1815C 3.7 0.34 5.3 4.5 14.0 44 0.904 0.2416B 2.0 0.14 2.5 3.1 5.8 20 0.427 0.2116C 3.5 0.31 2.7 2.9 8.0 30 0.592 0.17
1.57

Appendix 1.8. (continued)

Sample AVS Cd Cu Ni Pb Zn 3SEM 3SEM/A

18B 2.8 0.17 2.6 3.1 5.6 20 0.428 0.1518C 7.1 0.36 3.6 3.4 9.2 33 0.667 0.0919B 4.1 0.28 4.3 5.0 8.8 27 0.611 0.1519C 5.1 0.28 5.4 5.3 11.0 30 0.690 0.1420B 0.1 0.028 0.49 1.4 1.7 5.6 0.126 0.7920C 1.8 0.15 2.4 2.6 5.3 15 0.338 0.1921B 1.0 0.095 2.1 2.2 4.2 12 0.275 0.2821C 2.4 0.16 3.6 3.6 5.9 16 0.393 0.1622B 1.8 0.12 2.8 2.8 6.3 16 0.368 0.2022C 10.0 0.4 4.9 5.1 13.0 38 0.812 0.0824B 0.81 0.091 2.5 2.8 4.3 11 0.277 0.3424C 3.2 0.28 4.3 4.4 8.9 26 0.586 0.1825B 1.2 0.19 4.6 4.0 7.7 18 0.455 0.3825C 2.9 0.19 3.8 3.8 8.3 17 0.426 0.1526B 2.4 0.27 7.1 6.0 15.0 25 0.671 0.2826C 1.7 0.15 3.4 5.0 11.0 19 0.484 0.28SCB 1.7 0.31 9.0 3.4 17.0 35 0.820 0.48SCC 5.5 0.54 14.0 4.9 30.0 53 1.264 0.23
1.58

Appendix 1.9. Concentrations (μg/g dry weight) of organochlorine pesticides (OCs) in Upper Mississippi

River sediments.

POOL Chlordane Dieldrin DDE DDD DDT 

1B 0.001 ND 0.0004 0.0005 ND 1

1C ND ND ND ND ND

2B 0.001 0.0003 ND 0.0016 0.0002

2C ND ND 0.0520 0.0790 ND

3B ND 0.0003 0.0011 0.0038 0.0002

4B 0.002 0.0005 0.0010 0.0019 ND

4C ND ND ND ND ND

5B ND ND 0.0001 0.0001 ND

5C ND ND ND ND ND

6B ND ND 0.0001 0.0003 ND

6C ND ND ND ND ND

7B ND ND 0.0003 0.0010 0.0001

7C ND ND ND ND ND

8B ND ND 0.0002 0.0004 ND

8C ND ND ND ND ND

9B ND ND 0.0003 0.0010 0.0001

9C ND ND ND ND ND

10B ND ND 0.0002 0.0001 ND

10C ND ND ND ND ND

11B ND ND 0.0002 0.0004 ND

11C ND ND ND ND ND

12B ND ND 0.0003 0.0006 ND

12C ND ND ND ND ND

13B ND ND 0.0002 0.0004 ND

13C ND ND ND ND ND

14B ND 0.0001 0.0001 0.0002 ND

14C ND ND ND ND ND

15B 0.001 0.0002 0.0004 0.0005 0.0018

15C ND ND ND ND ND

16B ND 0.0002 0.0004 0.0004 ND

16C ND ND ND ND ND

18B ND 0.0002 0.0003 0.0006 ND

18C ND ND ND ND ND

19B ND 0.0003 0.0001 0.0002 ND

19C ND ND ND ND ND

20B ND ND 0.0001 0.0002 ND

20C ND ND ND ND ND

21B 0.002 0.0004 0.0004 0.0008 0.0003

21C ND ND ND ND ND
1.59

Appendix 1.9. Concentrations of organochlorine pesticides (OCs) in Upper Mississippi River sediments

(cont.). 

POOL Chlordane Dieldrin DDE DDD DDT 

22B ND 0.0003 0.0001 0.0001 ND

22C ND ND ND ND ND

24B 0.0010 0.0004 0.0001 0.0001 ND

24C ND ND ND ND ND

25B 0.0010 0.0006 0.0005 0.0005 ND

25C ND ND ND ND ND

26B ND 0.0007 0.0005 0.001 ND

26C ND ND ND ND ND

SCB ND ND 0.0007 0.0004 0.0001 

SCC ND ND 0.0780 0.0780 ND

ND = Not detected
1.60

Appendix 1.10. Concentrations ( μg/g dry weight) of polycyclic aromatic hydrocarbons (PAHs) in Upper Mississippi River sediments. (ND = Not Detected)

Pool 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 1B 1.080 ND ND 0.230 0.040 0.200 0.300 0.040 ND 0.600 0.580 0.190 0.280 0.390 ND N1C 0.013 0.013 ND ND ND ND 0.013 ND ND 0.031 0.025 0.031 0.031 0.038 0.038 0.2B 0.030 ND ND 0.020 0.020 0.020 0.230 0.020 ND 0.450 0.470 0.140 0.270 0.330 ND N2C 0.052 0.052 0.026 ND ND ND 0.157 0.026 0.052 0.340 0.314 0.157 0.209 0.157 0.183 0.3B 1.030 ND ND 0.380 ND 0.050 0.090 0.010 ND 0.210 0.190 0.060 0.140 0.210 ND N4B ND ND ND 0.590 ND ND 0.050 0.010 ND 0.350 0.420 0.050 0.140 0.230 ND N4C 0.049 0.049 ND ND ND ND 0.049 ND ND 0.196 0.245 0.098 0.147 0.147 0.147 0.5B ND ND ND 0.050 ND ND 0.010 ND ND 0.030 0.030 ND 0.010 0.020 ND N5C 0.036 0.036 ND ND ND ND ND ND ND 0.036 0.036 ND 0.071 0.036 ND N6B ND ND ND 0.010 ND ND 0.010 ND ND 0.090 0.100 ND 0.010 0.010 ND N6C 0.016 0.016 ND ND ND ND ND ND ND 0.031 0.031 ND 0.031 0.016 ND N7B ND ND ND 0.010 ND ND ND ND ND 0.110 0.130 ND 0.020 0.010 ND N7C 0.020 ND ND ND ND ND 0.020 ND ND 0.041 0.041 0.020 0.041 0.041 0.041 0.8B ND ND ND 0.040 ND ND 0.020 ND ND 0.040 0.030 0.010 0.020 0.020 ND N8C 0.019 0.019 ND ND ND ND 0.019 ND ND 0.037 0.019 0.019 0.037 0.019 0.019 0.9B 0.010 ND ND 0.010 ND 0.010 0.040 ND ND 0.080 0.070 ND 0.100 0.100 ND N9C 0.021 0.021 ND ND ND ND ND ND ND 0.042 0.021 0.021 0.042 0.021 0.042 0.10B ND ND ND 0.010 ND ND 0.010 ND ND 0.020 0.020 0.010 0.020 0.020 ND N10C 0.044 0.044 ND ND ND ND ND ND ND 0.044 0.044 ND 0.044 ND ND N

PAH-1 = Naphthalene PAH-2 = 2-methylnaphthalene PAH-3 = 1-methylnaphthalene PAH-4 = AcenaphthalenePAH-5 = Acenaphthene PAH-6 = Fluorene PAH-7 = Phenathrene PAH-8 = AnthracenePAH-9 =1,-methylphenanthrene PAH-10 =Fluoranthene PAH-11 = Pyrene PAH-12 = 1,2-BenzanthracenePAH-13 = Chrysene PAH-14 = Benzo(b)fluoranthene PAH-15 = Benzo(k)fluoranthene PAH-16 = Benzo(e)pyrenePAH-17 - Benzo(a)pyrene
1.61

Appendix 1.10. Concentrations of polycyclic aromatic hydrocarbons (PAHs) in Upper Mississippi River sediments (cont.).

Pool 18 19 20 21 22 23 24 25 26 27 28 29 30 31 32 33 1B ND 0.080 0.010 0.060 ND ND ND ND ND ND ND ND ND ND ND N1C ND ND ND ND 0.013 ND ND ND ND ND ND ND ND ND ND N2B ND 0.210 0.060 0.150 ND ND ND ND ND ND ND ND ND ND ND N2C 0.209 ND ND ND 0.079 ND 0.052 ND ND ND ND ND ND ND ND N3B ND 0.050 0.010 0.040 ND ND ND ND ND ND ND ND ND ND ND N4B ND 0120 0.020 0.080 ND ND ND ND ND ND ND ND ND ND ND N4C 2.304 ND ND ND 0.049 ND ND ND ND ND ND ND ND ND ND N5B ND ND ND ND ND ND ND ND ND ND ND ND ND ND ND N5C 1.286 ND ND ND 0.014 ND ND ND ND ND ND ND ND ND ND N6B ND ND ND ND ND ND ND ND ND ND ND ND ND ND ND N6C 0.094 ND ND ND 0.027 ND ND ND ND ND ND ND ND ND ND N7B ND ND ND ND ND ND ND ND ND ND ND ND ND ND ND N7C 1.660 ND ND ND ND ND ND ND ND ND ND ND ND ND ND N8B ND ND ND ND ND ND ND ND ND ND ND ND ND ND ND N8C 0.242 ND ND ND 0.022 ND ND ND ND ND ND ND ND ND ND N9B ND ND ND ND ND ND ND ND ND ND ND ND ND ND ND N9C 0.565 ND ND ND 0.019 ND ND ND ND ND ND ND ND ND ND N10B ND ND ND ND ND ND ND ND ND ND ND ND ND ND ND N10C 3.289 ND ND ND 0.013 0.026 ND ND ND ND ND ND ND ND ND N

PAH-18 = Perylene PAH-19 = Indeno(1,2,3-cd)pyrene PAH-20 = 1,2,5,6-dibenzanthracene PAH-21 = Benzo(g,h,i)perylenePAH-22 = C1-naphthalenes PAH-23 = C2-naphthalenes PAH-24 = C1-phenanthrenes PAH-25 = C2-phenanthrenesPAH-26 = C3-phenanthrenes PAH-27 = C4-phenanthrenes PAH-28 = C1-dibenzothiophenes PAH-29 = C2-dibenzothiophenesPAH-30 = C3-dibenzothiophenes PAH-31 = C1-fluoranthenes+C1-pyrene PAH 32 - C2-chrysenes PAH-33 = C3-chrysenesPAH-34 = C4-chrysenes
1.62

Appendix 1.10. Concentrations of polycyclic aromatic hydrocarbons (PAHs) in Upper Mississippi River sediments (cont.).

Pool 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 11B ND ND ND 0.230 ND ND 0.020 ND ND 0.090 0.080 0.010 0.030 0.040 ND N11C 0.016 0.016 ND ND ND ND ND ND ND ND ND ND ND ND ND N12B ND ND ND ND ND ND 0.010 ND ND 0.060 0.050 0.010 0.020 0.020 ND N12C 0.019 0.019 ND ND ND ND ND ND ND 0.019 0.019 ND 0.019 0.019 0.019 N13B ND ND ND 0.020 ND ND 0.010 ND ND 0.030 0.020 0.010 0.010 0.020 ND N13C 0.020 0.020 ND ND ND ND ND ND ND 0.020 0.020 ND 0.020 ND ND N14B ND ND ND 0.030 ND 0.040 0.020 ND ND 0.030 0.030 0.010 0.010 0.020 ND N14C 0.028 0.028 ND ND ND ND 0.028 ND ND 0.085 0.085 0.028 0.057 0.028 ND N15B ND ND ND 0.140 ND ND 0.070 ND ND 0.210 0.170 0.040 0.070 0.100 0.020 N15C 0.023 0.023 ND ND ND ND 0.023 ND ND 0.046 0.046 0.046 0.046 0.023 0.023 0.16B ND ND ND 0.050 ND 0.050 0.070 0.070 ND 0.140 0.120 0.030 0.070 0.090 ND N16C 0.015 0.015 0.015 ND ND ND 0.046 ND ND 0.091 0.107 0.091 0.091 0.061 0.076 0.18B ND ND ND 0.330 ND ND 0.020 ND ND 0.080 0.090 0.020 0.040 0.050 ND N18C 0.016 0.016 ND ND ND ND 0.033 ND ND 0.049 0.049 0.033 0.049 0.033 0.049 0.19B ND ND ND 0.180 ND 0.010 0.030 0.010 ND 0.060 0.070 0.02 0.040 0.060 ND N19C 0.021 0.021 ND ND ND ND 0.021 ND ND 0.064 0.064 0.043 0.064 0.043 0.043 0.20B ND ND ND 0.034 0.004 0.004 0.002 0.003 ND 0.033 0.030 0.012 0.010 0.001 ND N20C 0.014 0.014 ND ND ND ND 0.021 ND ND 0.043 0.043 0.043 0.058 0.043 0.029 0.

PAH-1 = Naphthalene PAH-2 = 2-methylnaphthalene PAH-3 = 1-methylnaphthalene PAH-4 = AcenaphthalenePAH-5 = Acenaphthene PAH-6 = Fluorene PAH-7 = Phenathrene PAH-8 = AnthracenePAH-9 =1,-methylphenanthrene PAH-10 =Fluoranthene PAH-11 = Pyrene PAH-12 = 1,2-BenzanthracenePAH-13 = Chrysene PAH-14 = Benzo(b)fluoranthene PAH-15 = Benzo(k)fluoranthene PAH-16 = Benzo(e)pyrenePAH-17= Benzo(a)pyrene
1.63

Appendix 1.10. Concentrations of polycyclic aromatic hydrocarbons (PAHs) in Upper Mississippi River sediments (cont.).

Pool 18 19 20 21 22 23 24 25 26 27 28 29 30 31 32 33 11B ND ND ND ND ND ND ND ND ND ND ND ND ND ND ND N11C 0.639 ND ND ND 0.016 ND ND ND ND ND ND ND ND ND ND N12B ND ND ND ND ND ND ND ND ND ND ND ND ND ND ND N12C 0.599 ND ND ND 0.019 ND ND ND ND ND ND ND ND ND ND N13B ND ND ND ND ND ND ND ND ND ND ND ND ND ND ND N13C 0.357 ND ND ND 0.020 ND ND ND ND ND ND ND ND ND ND N14B ND ND ND ND ND ND ND ND ND ND ND ND ND ND ND N14C 1.420 ND ND ND 0.028 ND ND ND ND ND ND ND ND ND ND N15B ND ND ND ND ND ND ND ND ND ND ND ND ND ND ND N15C 0.616 ND ND ND 0.023 ND ND ND ND ND ND ND ND ND ND N16B ND ND ND ND ND ND ND ND ND ND ND ND ND ND ND N16C 0.213 ND ND ND 0.030 ND ND ND ND ND ND ND ND ND ND N18B ND ND ND ND ND ND ND ND ND ND ND ND ND ND ND N18C 0.492 ND ND ND 0.016 ND ND ND ND ND ND ND ND ND ND N19B ND 0.010 0.010 0.010 ND ND ND ND ND ND ND ND ND ND ND N19C 0.596 ND ND ND 0.021 ND ND ND ND ND ND ND ND ND ND N20B ND ND ND ND ND ND ND ND ND ND ND ND ND ND ND N20C 0.261 ND ND ND 0.014 ND ND ND ND ND ND ND ND ND ND N

PAH-18 = Perylene PAH-19 = Indeno(1,2,3-cd)pyrene PAH-20 = 1,2,5,6-dibenzanthracene PAH-21 = Benzo(g,h,i)perylenePAH-22 = C1-naphthalenes PAH-23 = C2-naphthalenes PAH-24 = C1-phenanthrenes PAH-25 = C2-phenanthrenesPAH-26 = C3-phenanthrenes PAH-27 = C4-phenanthrenes PAH-28 = C1-dibenzothiophenes PAH-29 = C2-dibenzothiophenesPAH-30 = C3-dibenzothiophenes PAH-31 = C1-fluoranthenes+C1-pyrene PAH 32 - C2-chrysenes PAH-33 = C3-chrysenesPAH-34 = C4-chrysenes
1.64

Appendix 1.10. Concentrations of polycyclic aromatic hydrocarbons (PAHs) in Upper Mississippi River sediments (cont.).

POOL 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 21B ND ND ND 0.030 ND ND 0.070 0.010 ND 0.220 0.190 0.040 0.060 0.090 ND N21C 0.017 0.017 ND ND ND ND ND ND ND 0.017 ND ND 0.017 ND ND N22B ND ND ND 0.260 ND ND 0.020 ND ND 0.070 0.080 0.010 0.020 0.030 ND N22C 0.021 0.021 ND ND ND ND 0.021 ND ND 0.063 0.042 0.042 0.042 0.021 0.042 0.24B ND ND ND ND ND ND 0.010 ND ND 0.060 0.070 0.010 0.020 0.020 ND N24C 0.019 0.019 ND ND ND ND 0.019 ND ND 0.037 0.037 0.019 0.037 0.019 0.019 0.25B ND ND ND 0.010 ND ND 0.020 ND ND 0.050 0.050 0.020 0.030 0.040 ND N25C 0.018 0.018 ND ND ND ND ND ND ND 0.018 0.018 ND 0.037 ND ND N26B ND ND ND 0.010 ND ND 0.020 ND ND 0.050 0.050 0.020 0.030 0.050 ND N26C 0.014 0.014 ND ND ND ND ND ND ND 0.036 0.029 0.014 0.029 0.014 0.014 0.SCB ND ND ND 0.610 0.070 ND 0.050 0.010 ND 0.230 0.210 0.040 0.130 0.140 ND NSCC ND 0.039 ND ND ND ND ND ND ND 0.156 0.156 0.078 0.117 0.117 0.078 0.

PAH-1 = Naphthalene PAH-2 = 2-methylnaphthalene PAH-3 = 1-methylnaphthalene PAH-4 = AcenaphthalenePAH-5 = Acenaphthene PAH-6 = Fluorene PAH-7 = Phenathrene PAH-8 = AnthracenePAH-9 =1,-methylphenanthrene PAH-10 =Fluoranthene PAH-11 = Pyrene PAH-12 = 1,2-BenzanthracenePAH-13 = Chrysene PAH-14 = Benzo(b)fluoranthene PAH-15 = Benzo(k)fluoranthene PAH-16 = Benzo(e)pyrenePAH-17= Benzo(a)pyrene
1.65

Appendix 1.10. Concentrations of polycyclic aromatic hydrocarbons (PAHs) in Upper Mississippi River sediments (cont.).

POOL 18 19 20 21 22 23 24 25 26 27 28 29 30 31 32 33 21B ND ND ND ND ND ND 0.06 ND ND ND 0.03 ND ND ND ND 021C 0.087 ND ND ND 0.017 ND 0.017 ND ND ND ND 0.087 ND ND 0.017 N22B ND ND ND ND ND ND 0.02 0.03 ND ND 0.01 0.26 ND ND ND 022C 0.549 ND ND ND 0.021 ND 0.042 0.021 0.042 0.021 ND 0.549 ND ND 0.021 0.24B ND ND ND ND ND ND ND 0.02 ND ND ND ND ND ND ND 024C 0.595 ND ND ND 0.019 ND ND 0.019 0.019 0.019 0.037 ND ND ND ND 0.25B ND ND ND ND ND ND ND ND ND ND ND ND ND ND ND 025C 0.349 ND ND ND 0.018 ND ND 0.018 ND ND ND ND ND ND ND N26B ND ND ND ND ND ND ND ND ND ND ND 0.01 ND ND ND 026C 0.180 ND ND ND 0.014 ND ND ND ND ND ND ND ND ND ND NSCB ND 0.080 0.020 ND 0.050 ND ND ND ND ND ND 0.61 0.08 0.02 0.05 0SCC 5.469 ND ND ND 0.039 ND ND ND 0.078 0.078 0.117 ND ND ND ND N

PAH-18 = Perylene PAH-19 = Indeno(1,2,3-cd)pyrene PAH-20 = 1,2,5,6-dibenzanthracene PAH-21 = Benzo(g,h,i)perylenePAH-22 = C1-naphthalenes PAH-23 = C2-naphthalenes PAH-24 = C1-phenanthrenes PAH-25 = C2-phenanthrenesPAH-26 = C3-phenanthrenes PAH-27 = C4-phenanthrenes PAH-28 = C1-dibenzothiophenes PAH-29 = C2-dibenzothiophenesPAH-30 = C3-dibenzothiophenes PAH-31 = C1-fluoranthenes+C1-pyrene PAH 32 - C2-chrysenes PAH-33 = C3-chrysenesPAH-34 = C4-chrysenes
2.1

Chapter 2: An Evaluation of Bioaccumulation of Contaminants from Sediments from the Upper Mississippi

River Using Field-collected Oligochaetes and Laboratory-exposed Lumbriculus variegatus

Brunson, E.L., Canfield, T.J., Dwyer, F.J., Ingersoll, C.G., and Kemble, N.E.

Introduction

Over the past 10 years, a variety of methods have been described for evaluating the toxicity of sedimentassociated contaminants with benthic invertebrates. However, only a limited number of methods are currently

available for assessing bioaccumulation of contaminants from field-collected or laboratory spiked sediments

(Ingersoll et al 1995). Standard guides have been published for conducting 28-d bioaccumulation tests with the

oligochaete Lumbriculus variegatus including determination of bioaccumulation kinetics for different

compound classes (USEPA, 1994; ASTM 1996). Lumbriculus variegatus was selected for use in sediment

bioaccumulation testing in the present study of upper Mississippi River (UMR) for six reasons: (1) ease of

culture and handling, (2) known chemical exposure history, (3) adequate tissue mass for chemical analyses, (4)

tolerance of a wide range of sediment physico-chemical characteristics, (5) low sensitivity to contaminants

associated with sediment, and (6) amenability to long-term exposures without feeding. Other organisms do not

meet many of these selection criteria including mollusks (valve closure), midges (short-life cycle), mayflies

(difficult to culture), amphipods (small tissue mass, too sensitive), cladocerans and fish (not in direct contact

with sediment). 

Several investigators have conducted bioaccumulation studies in the laboratory with L. variegatus using

either field-collected or laboratory-spiked sediments (Schuytema et al. 1988; Nebeker et al. 1989; Ankley et al.

1991; Call et al., 1991; Carlson et al. 1991; Ankley et al. 1993; Kukkonen and Landrum 1994). However, only

one previous study has compared results of laboratory bioaccumulation studies conducted with L. variegatus to

residues from synoptically-collected field populations of oligochaetes (Ankley et al. 1992). The author reported

good agreement between concentrations of polychlorinated biphenyls in the laboratory and field organisms,

particularly for PCB congeners with K values <7. This suggests that laboratory exposures longer than 28 d ow

may be required to reach equilibrium for super-hydrophobic chemicals.

The United States Geological Survey (USGS) has been monitoring the Upper Mississippi River since 1987

to document the fate and transport of contaminated sediments (Moody and Meade 1995). Concern with the

redistribution of these contaminated sediments arose after the flood of 1993. This project is designed to

evaluate the current status of sediments in the UMR and is one chapter in a series designed to assess the extent

of sediment contamination in navigational pools of the river. The overall project consists of the following

assessments: (1) measuring concentrations of contaminants in sediments of the UMR (Moody et al. 1996), (2)

toxicity testing with sediments collected from the river (Chapter 1), (3) analysis of benthic community structure

(Chapter 3), and (4) bioaccumulation of sediment associated contaminants (the present chapter). The present

study had two objectives: (1) to assess the bioaccumulation of contaminants from UMR sediments using L.

variegatus and (2) to compare bioaccumulation in these laboratory-exposed oligochaetes to oligochaetes

collected from the field.

Materials and Methods

Sample Collection

Sediment samples and native oligochaetes were collected from 23 navigational pools on the UMR and from the

Saint Croix River ("C" samples described in Chapter 1). Sample stations were selected based on the potential of

oligochaetes or fine grained sediment. For each C sample, 35- to 80-L of sediment (6 to 25 grabs) were

collected with a stainless steel Ponar grab sampler (Wildlife Supply Company, Saginaw, MI). All grabs from a
2.2

station within a pool were collected within a 5-meter radius and combined in a 114-L high-density polyethylene

(HDPE) container. The composited sample was homogenized on board the research ship Acadiana using an

electric drill and a stainless steel auger. Once homogenized, the following subsamples were removed: (1) three

separate 250 ml subsamples for organic chemistry, metals/acid-volatile sulfides, and total organic

carbon/particle size (Chapter 1), (2) one 2-L subsample for benthic invertebrates (Chapter 3), and (3) one 10-L

subsample for laboratory toxicity (Chapter 1) and bioaccumulation testing. Sediment samples were stored at 4o

C until used in laboratory exposures or physical/chemical analysis.

The remainder of the composited C sample of sediment was rinsed on ship through a Wildco wash bucket (

U.S. Standard sieve size #30, 600 μm opening ). The material captured by the wash bucket was transferred to a

HDPE tub along with river water. After all the sediment was sieved, native oligochaetes were isolated from the

detritus. These oligochaetes from each sample were placed in a HDPE jar containing aerated river water and

held for 24 hours to depurate gut contents. After the 24-hour elimination period, dead oligochaetes were

discarded. The remaining oligochaetes were rinsed, blotted dry, weighed, transferred to clean glass jars, and

frozen at -22 C until analyzed for chemical contaminants. Weights of native oligochaete samples selected for o

analysis ranged from 0.34g (Pool 4) to 9.8g (Pool 9)

Laboratory Testing 

Lumbriculus variegatus were exposed in the laboratory to sediment following methods described in USEPA 

(1994) and ASTM (1996). Sediment from 13 of the 23 sampled pools were used in these laboratory exposures. 

Samples were chosen for testing on the basis of sufficient mass of field-collected oligochaetes for chemical

analyses (or the previously documented presence of PCBs for pool 4 in lower Lake Pepin; e.g. Rostad et al.,

1996 ). Oligochaetes were mass cultured in the laboratory following methods similar to those described in

USEPA (1994) using 75-L glass aquarium containing 50 L of well water (hardness 290 mg/L as CaCO , 3

alkalinity 255 mg/L as CaCO , pH 7.8). Each aquaria received about 27 volume additions (about 1.5 L/minute) 3

of well water daily. The culture water was aerated and maintained at 23 C. Pre-soaked, shredded brown paper o

towels were used as substrate. Cultures were fed Tetramin flake fish food twice weekly ad libitum.

Exposures of oligochaetes in the laboratory were conducted for 28 days in 4-L glass Pyrex beakers

containing 1 L of sediment and 3 L of overlying water. Four replicate chambers were tested for each of the

thirteen sediment samples. Reconstituted fresh water (hardness 90 to 96 mg/L as CaCO , alkalinity 60 to 70 3

mg/L as CaCO ; USEPA 1994) was used as the overlying water. Each beaker was calibrated to 4-L using a 3 

glass standpipe that exited through the beaker wall and was held in place with a silicon stopper. Test chambers

received 2 volume additions (6 L + 10% ) of overlying water per day. Water was delivered using a modified

Mount and Brungs diluter system (Ingersoll and Nelson 1990). An in-line flow splitter was attached to each

delivery line to split the water flow evenly to each of four test chambers. The splitters were constructed of 1/4

inch PVC pipe with four silicone stoppers and 14-gauge stainless steel hypodermic needles with the points and

connector ends cut off the needles (Figure 2.1). Glass stands were used to support the splitters keeping them

level to maintain a constant volume delivery to each exposure chamber. Chambers were held in a temperaturecontrolled waterbath (23+1 C) on a 16:8 light:dark photoperiod at about 500 lux. Oligochaetes were not fed o

during the sediment exposure.

Sediment and overlying water were placed in the chambers the day before adding organisms (Day -1). 

Sediments were first homogenized with a hand-held electric drill and stainless steel auger before being placed

into the test beakers. One-L of sediment was transferred into each chamber using a plastic spoon. Overlying

water was poured into the beakers through a piece of fine-mesh Nitex® material to minimize suspension of the

sediment. Water delivery started after chambers were placed in the waterbath.

Twenty-four hours before stocking the test (Day -1) oligochaetes were removed from the culture with a

fine-mesh nylon aquarium net, placed in beakers containing well water, and rinsed to remove excess toweling

and debris. Beakers containing the oligochaetes were then placed in a waterbath and aerated. With substrate
2.3

absent, the L. variegatus formed tight masses or clumps in the beakers which was helpful during transfer of

organisms into the exposure chambers. 

Oligochaetes were acclimated to the test water by removing half of the water in each beaker and replacing it

with temperature-acclimated test water. Two hours later this process was repeated. After another two hours, the

L. variegatus were combined into a glass pan and rinsed with well water to break up the masses of worms and

remove any remaining debris. With the mass of worms disturbed, oligochaetes were grouped together with a

stainless steel dental pick and allowed to form small clumps of about 1 g. The clumps of oligochaetes were

removed from the pan with the dental pick, touched against the rim of the pan to remove excess water, and

placed on a tared weigh boat. About 2.6 g unblotted oligochaetes were transferred to each test chamber

containing sediment and overlying water . Using this approach, the 2.6 g of unblotted oligochaetes represents

about 2 g of blotted oligochaetes or about 200 organisms. 

General conditions of the exposure system and behavior were evaluated daily. Dissolved oxygen and

conductivity of the overlying water were measured weekly in all chambers. Total hardness (as CaCO ), pH, 3

alkalinity (as CaCO ), and total ammonia of overlying water were measured at the beginning and end of the test. 3

Overlying water pH, alkalinity, total hardness, conductivity and total ammonia measurements were similar

among all stations and inflowing test water (Appendix 2.1). Dissolved oxygen measurements were at or above

acceptable levels (>40% of saturation; ASTM 1996) in all treatments throughout the study (Appendix 2.1). 

Ranges of mean water quality for each parameter were as follows: pH 7.7 to 7.9; alkalinity as CaCO 61 to 67 3 

mg/L; total hardness as CaCO3 104 to 110 mg/L; conductivity 342 to 350 μS @25 C; total ammonia 0.1 to 0.4 o

mg/L; and calculated unionized ammonia 0.0028 to 0.0094 mg/L. 

On Day 28 of the exposure, L. variegatus were isolated from each test chamber by washing the sediment

through No. 18 (1.0 mm opening) followed by No. 50 (300 μm opening) U.S. standard stainless steel sieves. 

The contents of each sieve was rinsed into several clear glass pans and all oligochaetes were removed. 

Lumbriculus variegatus were separated from native oligochaetes based on behavior (native oligochaetes tended

to form a tight, spring-like coil, whereas L. variegatus would not (USEPA 1994)). Once isolated, all L.

variegatus from a chamber were cleaned of any remaining debris and held for 24 h in 1-L water-only chambers

to allow them to clear their gut contents. The L. variegatus were then isolated, cleaned of any remaining debris,

and transferred to a tared weigh boat. Samples were then blotted, weighed, placed in glass jars, and stored at -

22 C pending chemical analysis for contaminants. Weights of laboratory-exposed oligochaete samples ranged o

from 1.3g to 3.0g.

Chemical Analyses

Sediment physical characteristics included the following: (1) sediment particle size, (2) total organic carbon, (3)

inorganic carbon and (4) percent water. Sediment chemical parameters included: (1) organochlorine pesticides

(OCs), (2) polychlorinated biphenyls (PCB), (3) select aliphatic and polynuclear aromatic hydrocarbons (PAH),

(4) simultaneously extracted metals (SEM), (5) acid volatile sulfide (AVS), and (6) total metals. See Chapter 1

for additional information on methods and results of chemical and physical characterizations of the sediments. 

Concentrations of metals and organochlorines in sediment samples were low (Chapter 1). Therefore,

replicate tissue samples from the laboratory exposures were combined for organochlorine pesticide/PCB

analyses and metals were not analyzed because of limited sample mass. Tissues were analyzed by Geochemical

and Environmental Research Group at Texas A&M University, College Station, Texas for the following: (1)

organochlorine pesticides (OCs), (2) polychlorinated biphenyls (PCBs), (3) select aliphatic and polynuclear

aromatic hydrocarbons (PAHs), and (4) percent lipid. Prior to analysis, tissue samples were homogenized and

extracted using a Teckmar Tissumizer, sodium sulfate, and methylene chloride (MacLeod et al. 1985; Wade et

al. 1988; Brooks et al. 1989). Tissue extracts were split into two fractions: one fraction was used to measure

percent lipid and the second fraction was used for measuring PAHs, OCs, and PCBs. Extracts for chemical

analyses were purified using absorption chromatography to isolate the aliphatic fraction and the PAH/OC/PCB
2.4

fraction. Lipid interference in the PAH/OC/PCB fraction was eliminated with further purification using HPLC. 

The quantitative analyses were performed by capillary gas chromatography (CGC) with electron capture

detector for OCs and PCBs and a mass spectrometer detector in the SIM mode for PAHs (Wade et al., 1988). 

Percent lipids were calculated on a wet-weight basis. A 20-ml aliquot of the total extract was filtered,

concentrated to 1 ml, and weighed. A 100-ul subsample was then removed, evaporated to dryness, and

weighed. Percent lipid was calculated using the weight of the dried subsample and the concentrated sample. 

Tissue residue data are presented in Appendix 2.2 and Appendix 2.3. Sediment data are shown in Table 1.1,

and Tables 1.3 to 1.5 in Chapter 1.

Average percent spike recovery for twenty-two OCs and was 88% (n=4). Beta BHC had the smallest

average spike recovery (53%) while oxychlordane had the greatest average spike recovery (104%). Individual

OC concentrations were often below minimum detectable limits so duplicate analyses were evaluated only for

total PCBs. The average duplicate coefficient of variation was 26% (range 0.7 to 61%, n=4). Average percent

spike recovery for PAH compounds was 96% (25 compounds, n=4). L123(c,d)pyrene had the smallest average

percent recovery (81%) while 1-methylnaphthalene had the greatest average percent recovery (110%). The

average duplicate coefficient of variation was 21% (34 possible compounds, n=1-4). Average duplicate

coefficient of variation ranged from 1% for c1-phenanthracene to 79% for benzo-a-pyrene. 

In addition to the laboratory-exposed and field-collected oligochaetes, three samples of oligochaetes from

laboratory cultures were collected at the beginning of the exposure for analysis contaminants. Two of the three

samples had detectable concentrations of PAHs and total PCBs however, the concentrations were generally less

than those of oligochaetes exposed to or collected from the UMR sediments. For some unexplained reason,

total PCB (1.3 μg/g dry wt ) and some PAH concentrations (up to 0.25 μg/g dry wt. ) in one of those three

samples was similar to oligochaetes exposed during the test. 

Results and Discussion

General Trends

Individual organochlorine pesticides (OC) were generally below the detection limits (ranging from 0.0007

to.0.0217 μg/g wet weight) for oligochaetes from both field-collection and laboratory-exposed animals

(Appendix 2.2). For the 13 field collected samples and 22 OCs measured, individual OCs were identified a total

of 6 times. The greatest individual OC concentration was 0.009 μg/g (wet weight) for dieldrin from

oligochaetes collected from Pool 22. As was the case with the field-collected oligochaetes, tissue

concentrations of individual OCs were often below the detection limit for many of the laboratory-exposed

oligochaetes. All oligochaete samples had at least one OC concentration above background (Pool 13 and Pool

16; 4,4'DDE); however, no sample had more than 6 OCs detected (Pool 11 and 14; gamma-chlordane, alphachlordane, aldrin, dieldrin, 4,4'DDE, 4,4'DDD). The greatest individual OC concentration was 0.013 μg/g (wet

weight) for 4,4 DDE for oligochaetes exposed in the laboratory to sediment collected from Pool 4. Also, 4,4

DDE was the most frequently measured OC (12 samples) with concentrations ranging from 0.0021 to 0.013

μg/g (wet weight).

Total PCBs were the only chlorinated organic compound detected in all field-collected and laboratoryexposed oligochaetes. Concentrations ranged from 0.045 μg/g (wet weight - pool 13) to 0.697 μg/g (wet weight

- Pool 4). The geometric mean for total PCBs measured in oligochaetes exposed to the sediment samples was

0.129 μg/g 

Field-collected and laboratory-exposed oligochaete samples were analyzed for 44 PAH isomers. Field

collected oligochaetes from Pool 4 had the fewest number of PAHs (14) while Pool 19 had the most (36). Only

16 PAH isomers (about 40% of those analyzed for) had detectable concentrations (detection limits from 0.0217

to 0.0024 μg/g wet weight) in 7 of the 13 Pools for both the field-collected and laboratory-exposed oligochaetes

(for the laboratory exposures, 2 of the 4 replicates had to exceed the detectable limit in order to be included in
2.5

this analysis). Table 2.1 lists all compounds measured in tissues that met these selection criteria. Figures 2.2

and 2.3 depict accumulation of total PAH in samples from laboratory-exposed or field-collected oligochaetes

for each UMR pool evaluated. Concentrations of the 16 PAH isomers were converted to molar units,

normalized to percent lipid, and summed. Total PAH from field-collected and laboratory-exposed

oligochaetes, show a trend of decreasing concentrations in the down river Pools (14 to 22). Field-collected

oligochaetes from Pool 7 were more contaminated than oligochaetes from the other pools. For the laboratory

exposures, oligochaetes exposed to sediments from Pool 4 were more contaminated than oligochaetes exposed

to sediments from the other pools. In general, perylene had the highest concentration of any PAH from fieldcollected and laboratory-exposed oligochaetes. This trend was greater for laboratory exposed oligochaetes than

for those collected from the field. Perylene concentrations ranged from 0.056 to 0.53 μg/g (wet weight) in field

collected oligochaetes and from 0.052 to 0.84 μg/g (wet weight) in oligochaetes from laboratory exposures.

Sediments and oligochaetes from the UMR are relatively uncontaminated compared to other locations we

have evaluated using sediment toxicity tests (Ingersoll et al. 1996) or bioaccumulation tests (sediments from

Little Scioto River in Ohio, unpublished data). Ingersoll et al (1996) calculated sediment effect concentrations

including Effects Range Medians (ERMs) from 28-day sediment exposures with Hyalella azteca. An ERM is 

defined as that concentration of a material in sediment above which toxic effects are frequently or always

observed or predicted. In the current study, tissue concentrations of PAHs were generally greatest in samples

from Pool 4. Two low molecular weight (LMW) PAHs (naphthalene and phenanthrene) and two high

molecular weight (HMW) PAHs (pyrene and chrysene) were generally the PAHs of highest concentration in

tissue samples from pool 4. The calculated sediment ERM concentrations (ug/g dry weight) for those PAHs are;

naphthalene - 0.097 , phenanthrene - 0.345, pyrene - 0.347, and chrysene - 0.500. The sediment concentrations

(ug/g dry weight) from Pool 4 were; naphthalene - 0.049, phenanthrene- 0.049, pyrene - 0.245, and chrysene -

0.147. The sediment ERMs are 1.4 to 7 times greater than the highest concentrations of these PAHs in

sediments from the current study. ERMs are not directly applicable to contaminant concentrations in tissues;

however, tissue concentrations in UMR Pool 4 were more than two orders of magnitude less than tissue

concentrations of oligochaetes exposed to sediments from the Little Scioto River. Collectively, this information

would indicate that sediment and biota from the UMR is relatively uncontaminated when compared to known

contaminated sites previously evaluated by our laboratory.

Detection of Compounds in Tissue vs. Sediment

Detection limits for tissue and sediment are usually different which creates difficulties in interpreting

bioaccumulation potential from relatively uncontaminated sediments. In the UMR, concentrations of PAHs and

PCBs were detected in both sediments and tissue samples 79% of the time for the laboratory-exposed

oligochaetes and 58% of the time for the field-collected oligochaetes. PAHs and PCBs were not detected in the

sediments but were detected in laboratory-exposed oligochaetes in 17% of the samples and in field-collected

oligochaetes in 41% of the samples. PAHs and PCBs were detected in sediment samples but not in 3% of the

samples from laboratory-exposed oligochaetes and 1% of the samples of field-collected oligochaetes. Although

the detection limits for sediments and tissues met established guidelines (USEPA 1984), detection limits for

sediments may need to be decreased in order to better represent potentially bioavailable compounds. 

Laboratory to Field Comparisons

Tissue concentrations of naphthalenes were generally higher in field-collected oligochaetes than in laboratory

exposed oligochaetes (Figure 2.4). Naphthalenes are LMW PAHs with log K values less than 4.5. PAHs with ow 

similar concentrations in both the laboratory-exposed and field-collected oligochaetes included a similar number

of HMW and LMW compounds (biphenyl, fluorene, 1-methylphenanthrene, pyrene, fluoranthene, chrysene, and
2.6

benzo(e)pyrene). Most of these compounds are intermediate in molecular weight and log K (except for ow

benzo(e)pyrene which has the highest molecular weight and log Kow of all compounds included in Figure 2.4). 

PAHs typically higher in the laboratory-exposed than in field-collected oligochaetes were primarily HMW

compounds ( benzo(a)anthracene, benzo[b(k)]fluoranthene, and perylene) with log K s greater than 5.1 (Figure ow

2.4 and 2.5). 

The ratio of tissue concentrations in laboratory-exposed oligochaetes to concentrations in field-collected

oligochaetes were generally similar (Figure 2.5). About 90% of the corresponding concentrations were within a

factor of three between the laboratory-exposed and field collected oligochaetes (represented by the crosshatched

region in Figure 2.5). However, there appears to be a shift from field>lab to lab>field as the molecular weight

of PAHs increases. Concentrations that differed by more than a factor of three were primarily LMW PAHs

(naphthalene, 1-methylnaphthalene, 2-methylnaphthalene, 2,6-dimethylnaphthalene, fluorene, 1,6,7-

trimethylnaphthalene, phenanthrene, and 1-methylphenanthrene) and were usually elevated in the field-collected

oligochaetes compared to the laboratory-exposed oligochaetes. Ratios >3 in the laboratory-exposed or fieldcollected oligochaetes were most frequently associated with a small group of pools (Field > 3x lab in Pools 4,

12, 22; lab >3x field in Pool 7).

Differences between tissue concentrations in the laboratory-exposed and field-collected oligochaetes may

have resulted from LMW PAHs being lost during the sampling of sediments. A second possibility for

differences between the laboratory and field-exposed may be spatial heterogeneity of contaminants in the

sediments in the field. Other possible explanations could include the rout of exposure. Exposure to

contaminants in the field may occur through sediment, food and overlying water while the route of exposure to

oligochaetes in the laboratory was sediment. Species-specific differences in exposure between Lumbriculus

variegatus and the native oligochaetes may also contribute to the differential accumulation. For example,

concentrations of metals reportedly differ among taxa inhabiting the same locations (Cain et al. 1992).

Biota-sediment Accumulation Factors

Biota-sediment accumulation factors (BSAFs) were calculated by dividing the lipid-normalized tissue

concentrations by the organic-carbon normalized sediment concentrations (USEPA 1994). Mean BSAFs for

this study were only listed for compounds in which BSAF could be calculated for both laboratory-exposed and

field-collected oligochaetes in at least seven of 13 pools (Table 2.2). For laboratory-exposed oligochaetes, mean

BSAFs ranged from 1.1 for benzo(a)anthracene to 5.3 for naphthalene. Mean BSAFs for field-collected

oligochaetes, mean BSAFs ranged from 0.5 for benzo(a)anthracene to 8.8 for naphthalene. Individual sample

BSAFs for naphthalene ranged from 1.6 to 10.1 in laboratory-exposed oligochaetes and 2.5 to 26.6 in fieldcollected oligochaetes. BSAFs for pyrene, benzo(a)anthracene, and benzo(b,k)fluoranthene were typically

greater than BSAFs reported for marine organisms (Lee 1992). BSAFs were also calculated using PCB

homolog data reported in Ankley et al. (1992) for laboratory-exposed L. variegatus and field-collected

oligochaetes (Figure 2.6). BSAFs were similar between laboratory-exposed and field-collected oligochaetes in

both Ankley et al. (1992) and in the present study; however, BSAFs in the present study were typically greater

(0.5 to 8.8) than those from Ankley et al. (1992; 0.17 to 2.26). 

A theoretical value of 1.7 for BSAFs has been estimated based on partitioning of non-ionic organic

compounds between sediment carbon and tissue lipids (McFarland and Clarke 1986). A BSAF of less than 1.7

indicates less partitioning into lipids than predicted and a value greater than 1.7 indicates more uptake than can

be explained by partitioning theory alone (Lee 1992). The majority of the BSAFs in Table 2.2 were within a

range of about 0.5 to 2.6 suggesting the theoretical BSAF value of 1.7 could be used to predict these mean

BSAFs with a fair amount of certainty. However, mean BSAFs for naphthalene (8.8) and 2-methyl naphthalene

(6.7) in the field-collected oligochaetes were elevated relative to a theoretical BSAF of 1.7. Moreover, BSAFs

for individual pools were as high as 10.1 for laboratory-exposed oligochaetes and 26.6 for field-collected

oligochaetes. The higher BSAFs in the field-collected oligochaetes may be the result of (1) exposure to
2.7

contaminants in the overlying water, (2) spatial differences in sediment contamination (i.e., sediments were not

sampled from a depth representative of the habitat of the oligochaetes), (3) increased error in chemical

determinations due to low concentration of contaminants in sediments, or (4) taxonomic-specific differences in

exposure. BSAFs substantially different from the theoretical value of 1.7 may also result when the system has

not reached steady state (i.e., depletion or release of contaminants in pore water).

Summary

Contaminant concentrations were relatively low in sediments and tissues from the 13 UMR pools evaluated. 

Only PAHs and total PCBs were frequently measured above detection limits. Most of the concentrations of

PAHs in UMR sediment were similar to concentrations in sediments identified as non-toxic in amphipod

toxicity tests from these previous studies. PAH concentrations in tissues of oligochaetes tested with highly

contaminated samples from previous studies were up to 1000 times greater than tissue concentrations measured

in the present study. Concentrations in laboratory exposed and field-collected oligochaetes for a compound

from a specific pool in the UMR were generally similar. About 90% of the paired PAH concentrations in

laboratory-exposed and field-collected oligochaetes were within a factor of three of one another. With the

detection limits used to analyze samples in the present study, contaminants were detected in tissue samples more

often than in the associated sediment samples. 

Acknowledgments. We would like to thank the following individuals for their contributions to the project: I.E.

Greer, D.K. Hardesty, P.S. Haverland, C.E. Henke, E.K. Henry, P. A. Lovely, J.A. Moody, D.S. Reussler, J.D.

Soltvedt, J. Stevens, R.W. Walton, D.W. Whites, P.R. Heine, J.L. Zajicek, D.C. Zumwalt, and the Crew of the

Acadiana (Craig LeBoeuf and Pat Marmande).

References

Ankley GT, Benoit DA, Hoke RA, Leonard EN, West CW, Phipps GL, MattsonVR, 

Anderson LA (1993) Development and Evaluation of Test Methods for Benthic Invertebrates and

Sediments: Effects of Flow Rate and Feeding on Water Quality and Exposure Conditions. Arch Environ

Toxicol Chem. 25:12-19

Ankley GT, Lodge K, Call DJ, Balcer MD, Brooke LT, Cook PM, Kreis Jr.RG, Carlson AR, 

Johnson RD, Niemi GJ, Hoke RA, West CW, Giesy JP, Jones PD, Fuying ZC (1992) Intergrated

assessments of contaminated sediments in the lower Fox River and Green Bay, Wisconsin. Ecotoxicol

Environ Saf 23:46-63 

Ankley GT, Schubauer-Berigan MK, Dierkes JR (1991) Predicting the toxicity of bulk 

sediments to aquatic organisms using aqueous test fractions: pore water versus elutriate. Environ. Toxicol.

Chem. 10:1359-1366

American Society for Testing and Materials (ASTM) (1996) Standard guide for determination of 

bioaccumulation of sediment-associated contaminants by benthic invertebrates. E1688-95. Annual Book of

ASTM Standards, Vol. 11.05, Philadelphia, PA. pp. 1110-1159

Brooks JM, Wade TL, Atlas EL, Kennicutt II MC, Presley BJ, Fay RR, Powell EN, Wolf G 

(1989) Analysis of bivalves and sediment for organic chemicals and trace elements. Third annual Report

for NOAA’s National Status and Trends Program, Contract 50-DGNC-5-00262
2.8

Cain DJ, Luona SN, Carter JL, Fend SV (1992) Aquatic insects as bioindicators of trace 

element contamination in cobble-bottom rivers and streams. Can.J. Fish. Aquat. Sci. 49:2141-2154

Call DJ, Balcer MD, Brooke LT, Lozano SJ, Vaishnav DD (1991) Sediment quality 

evaluation in the lower Fox River and southern Green Bay of Lake Michigan. USEPA Cooperative

Agreement Final Report, University of Wisconsin-Superior, Superior, WI

Carlson AR, Phipps GL, Mattson VR, Kosian PA, Cotter AM (1991) The role of acidvolatile sulfide in determining cadmium bioavailability and toxicity in fresh-water sediments. Environ.

Toxicol. Chem. 14:1309-1319

Ingersoll CG, Nelson MK (1990) Testing sediment toxicity with Hyalella azteca (Amphipoda) 

and Chironomus riparius (Diptera). In: Landis WG, van der SchalieWH (eds) Aquatic Toxicology and

Risk Assessment: Thirteenth Volume. STP 1096. American Society for Testing and Materials, 

Philadelphia, PA, pp. 93-109

Ingersoll CG, Ankley GT, Benoit DA, Burton GA, Dwyer FJ, Greer IE, Norberg-King TJ, 

Winger PV(1995) Toxicity and bioaccumulation of sediment-associated contaminants with freshwater

invertebrates: A review of methods and applications. Environ. Toxicol. Chem. 14:1885-1894 

Ingersoll CG, Haverland PS, Brunson EL, Canfield TJ, Dwyer FJ, Henke CE, Kemble NE, 

Mount DR, Fox RG (1996) Calculation and evaluation of sediment effect concentrations for the amphipod

Hyalella azteca and the midge Chrinomous riparius. J Great Lakes Res. 22(3):602-623

Kemble NE, Brunson EL, Canfield TJ, Dwyer FJ, Ingersoll CG (1996) Evaluation of 

contamination in sediments collected from navigational pools of the Upper Mississippi River using a 28

day Hyalella azteca test. (Chapter 1)

Kukkonen J, Landrum PF (1994) Toxicokinetics and toxicity of sediment bound pyrene in 

Lumbriculus variegatus (Oligochaeta). Environ Toxicol Chem 13:1457-1468 

Lee H II (1992) Models, muddles and mud:... In: Burton GA (ed) Sediment Toxicity Assessment.

Lewis Publishers, Chelsea MI pp 267-293

MacLeod WD, Brown DW, Friedman AJ, Burrow DG, Mayes O, Pearce RW, Wigren CA, 

Bogar RG (1985) Standard analytical procedures of the NOAA National Analytical Facility 1985-1986. 

Extractable Toxic Organic Compounds. 2nd Ed. U.S. Department of Commerce, NOAA/NMFS. NOAA

Tech. Memo. NMFS F/NWC-92. 

McFarland VA, Clarke JU (1986) Testing bioavailability of polychlorinated biphenyls using a 

two-level approach. In: Wiley RG (ed) Water Quality Research and Development: Successful Bridging

between Theory and Application. Hydrologic Engineering Research Center, Davis, CA, pp. 220-229
2.9

Moody JA, and Meade RH (1995) Hydrologic and sedimentologic data collected during three 

cruises on the Mississippi River and some of its tributaries from Minneapolis, Minnesota, to New Orleans,

Louisiana, July 1991-May 1992: U.S. Geological Survey Open-File Report 94-474 

Moody JA (1996) Hydrologic and sedimentologic, and chemical data describing surficial bed 

sediments in the navigational pools of the Upper Mississippi River after the Flood of 1993. U.S.Geological

Survey Open File Report 96-580. J.A. Moody, editor

Nebeker AV, Griffis WL, Wise CM, Hopkins E, Barbitta JA (1989) Survival reproduction 

and bioconcentration in invertebrates and fish exposed to hexachlorobenzene. Environ Toxicol Chem 

8:601-611

Rostad CE, Bishop LM, Ellis GS, Leiker TH, Monsterleet SG, Pereira WE (1986) 

Polychlorinated biphenyls and other synthetic organic contaminants associated with sediments and fish in

the Mississippi River. In: Contaminants in the Mississippi River 1978-92. U.S. Geological Survey Circular

1133:103-113

Schuytema GS, Krawczyk DF, Griffis WL, Nebeker AV, Robideaux ML, Brownawell BJ, 

Westall, JC (1988) Comparative uptake of hexachlorobenzene by fathead minnows, amphipods and

oligochaete worms from water and sediment. Environ Toxicol Chem 7:1035-1044

United States Environmental Protection Agency (1994) Methods for Measuring the Toxicityand 

Bioaccumulation of Sediment-associated Contaminants with Freshwater Invertebrates. EPA/600/R-94/024.

Washington, DC.

United States Environmental Protection Agency (1984) Guidelines Establishing Test Procedures 

for the Analysis of Pollutants Under the Clean Water Act; Final Rule and Interim Final Rule and Proposed

Rule. USEPA Federal Register Part VII 40 CFR Part 136.

Wade TL, Atlas EL, Brooks JM, Kennicutt II MC, Fox RG, Sericano J, Garcia B, DeFreitas 

D (1988) NOAA Gulf of Mexico Status and Trends Program: Trace Organic Contaminant Distribution in

Sediments and Oysters. Estuaries 11:171-179
2.10

Fig. 2.1. Diagram of in-line flow splitter used to deliver overlying water.
4

5

6

7

9

10

11

12

13

14

16

19

22

Sample number

0001111

uMole/g lipid

2.11

Fig. 2.2. Total accumulation of polycyclic aromatic hydrocarbons (μMole/g lipid) by Lumbriculus variegatus exposed in from the Upper Mississippi River. Chemical numbers correspond to the following chemicals: (1) naphthalene, (2) 1-memethylnaphthalene, (4) biphenyl, (5) 2,6-dimethylnaphthalene, (6) fluorene, (7) 1,6,7-trimethylnaphthalene, (8) phenanthmethylphenanthrene, (10) pyrene, (11) fluoranthene, (12) chrysene, (13) benzo(a)anthracene, (14) benzo[b(k)]fluoranthebenzo(e)pyrene.
4

5

6

7

9

10

11

12

13

14

16

19

2

Sample number

0001111

uMole/g lipid

2.12

Fig. 2.3. Total accumulation of polycyclic aromatic hydrocarbons (μMole/g lipid) by oligochaetes collected from select p

Mississippi River. Chemical numbers correspond to those listed for Figure 2.2.
0

5

10

FIELD

05

10

LAB

BIPHENYL

0

1

2

3

4

5

6

7

8

FIELD

012345678

LAB

1-METHYLNAPHTALENE

0

1

2

3

4

5

6

7

8

FIELD

012345678

LAB

2-METHYLNAPHTALENE

0

5

05

101520

LAB

2,6-DIM

0

2

4

6

8

10

FIELD

02468

10

LAB

1,6,7-TRIMETHYLNAPHTALENE

A

A

A

A

A

B

0

5

05

1015202530

LAB

NA

2.13

Fig 2.4. Comparison of tissue concentrations in laboratory-exposed L. variegatus verses field-collected oligochaetes. An 

"B" indicates laboratory = field, and "C" indicates laboratory > field.
0

1

2

3

4

5

6

7

8

9

10

11

12

13

123456789

10

LAB > FIELD

0

1

2

3

4

5

6

7

8

9

10

11

12

13

123456789

10

FIELD > LAB

COMPOUND #

A AABA B ACCB BBCC 

2.14

Fig. 2.5. Ratio of tissue concentrations in laboratory-exposed or field-collected oligochaetes for select PAHs. See the leglisting of the specific compounds by number. An "A" indicates field > laboratory, "B" indicates laboratory similar to fiellaboratory > field. Compounds are plotted in order of molecular weight with molecular weight increasing from left to rigconcentration of a compound for a pool is higher than the corresponding field concentration, then the laboratory/field ratof the plot. If the field concentration of a compound for a pool is higher than the corresponding laboratory concentrationratio is plotted on the lower half of the plot (see Appendix 2.4 for a list of ratio values).
0 1 2 3 4 5 6 7 8 

 Field-collected oligochaetes

0123456

Laboratory-exposed Lumbriculus

2.15

Fig 2.6. Biota-sediment accumulation factors (BSAFs) for laboratory-exposed Lumbriculus variegatus and field-collecte

the present study (circles) and calculated from PCB homolog data reported in (Ankley et al. 1992;triangles).
2.16

Table 2.1. List of chemicals that met our criteria for laboratory to field comparisons of tissue concentrations and their aand log Kow.

Chemical No. Low molecular-weight PAHs Molecular Weight Log Kow 

1 NAPHTHALENE 128.17 3.35 2 1-METHYLNAPHTHALENE 142.20 3.87 3 2-METHYLNAPHTHALENE 142.20 4.00 4 BIPHENYL 154.21 3.90 5 2,6-DIMETHYLNAPHTHALENE 156.23 4.31 6 FLUORENE 166.22 4.38 7 1,6,7-TRIMETHYLNAPHTHALENE 170.25 8 PHENANTHRENE 178.23 4.57 9 1-METHYLPHENANTHRENE 192.26 5.14 

High Molecular-weight PAHs

10 PYRENE 202.26 5.18 11 FLUORANTHENE 202.26 5.22 12 CHRYSENE 228.29 5.86 13 BENZOaANTHRACENE 228.29 5.91 14 BENZO[b(k)]FLUORANTHENE 252.32 5.78, 6.20 15 PERYLENE 252.32 6.25 16 BENZOePYRENE 252.32 6.44 
2.17

Table 2.2. Biota-sediment accumulation factors reported by Lee (1992) and in the present study. The meas BSAFs for t

where there was matching detection of a particular compound in both sediment and tissue in a least seven on 13 pools fofield collected (field) oligochaetes. The range in BSAFs in the present study are reported for as values for samples from upper Mississippi River.

COMPOUND Lee (1992) RANGE LAB RANGE FIELD RANGENaphthalene 5.3 1.6-10.1 8.8 2.5-26.62-methylNaph. 2.6 0.9-5.1 6.7 2.2-12.2Pyrene 0.4 0.18-0.5 2.3 0.8-3.9 2.2 0.7-5.6Fluoranthene 1.8 0.9-3.9 1.6 0.6-4.9Chrysene 1.5 0.7-2.4 1.1 0.3-2.0Benzo(a)anthracene 0.4 0.2-0.6 1.1 0.4-2.5 0.5 0.4-0.7Benzo[b,(k)]fluoran. 0.4 0.2-1.0Perelene 2.24 0.5-4.7 1.02 0.3-1.9
2.18

Appendix 2.1. Mean measured overlying water quality for the laboratory bioaccumulation test with Upper Mississippi River samples.

Pool Dissolved Oxygen Temperature Conductivity pH Alkalinity Hardness To

(mg/L) C (uS) (mg/L CaCO3) (mg/L CaCO3) (m

4 6.8 22.4 344.2 7.9 63.0 109.5 5 6.3 22.3 346.2 7.8 62.0 104.0 6 6.2 22.3 347.0 7.8 65.5 105.0 7 6.7 22.3 341.6 7.8 64.0 105.5 9 6.8 22.1 348.2 7.9 65.0 109.0 

10 6.5 22.3 348.8 7.8 63.0 107.5 11 6.4 22.5 349.2 7.8 61.0 106.5 12 5.8 22.5 347.0 7.7 66.5 105.0 13 6.5 22.3 344.2 7.8 64.0 106.5 14 6.9 22.4 345.2 7.8 65.0 107.0 16 6.0 22.5 350.0 7.8 66.0 109.5 19 6.2 22.5 347.2 7.8 67.0 107.0 22 6.0 22.3 350.3 7.7 65.0 110.0 

Mean 6.4 22.4 346.9 7.8 64.4 107.1 

Max. 6.9 22.5 350.3 7.9 67.0 110.0 

Min. 5.8 22.1 341.6 7.7 61.0 104.0 

Std. 0.4 0.1 2.5 0.1 1.8 1.9 

Median 6.4 22.3 347.0 7.8 65.0 107.0 
2.19

Appendix 2.2. Tissue concentrations of organochlorine compounds measured in laboratory-exposed and field-colconcentrations are on a wet-weight basis (μg/g). Appendix 2.2 data can be obtained electronically from:

anonymous ftp - ftp://ftp.msc.nbs.gov/pub/umr/umr.zipworld wide web - http://www.msc.nbs.gov/pubs/umr.html

For problems with access to the above addresses please e-mail the Webmaster, Chris Henke, at chenke@msc.nbs.
2.20

Appendix 2.3. Tissue concentrations of PAHs measured in laboratory-exposed and field-collected oligochaetes. Awet-weight basis (μg/g). Appendix 2.3 data can be obtained electronically from:

anonymous ftp - ftp://ftp.msc.nbs.gov/pub/umr/umr.zipworld wide web - http://www.msc.nbs.gov/pubs/umr.html

For problems with access to the above addresses please e-mail the Webmaster, Chris Henke, at chenke@msc.nbs.
2.21

Appendix 2.4 Total accumulation (μMole/g lipid) of PAHs in Laboratory exposed (LMML) and Field Collected (Lipid-normalized concentrations (μg/g lipid) are given for laboratory-exposed (LLCONC) and field-collected (FLChemical numbers (CHEM) correspond to those listed in Figure 2.3. 

 OBS POOL LLCONC FLCONC MOLEWT CHEM LMML FMML

 1 4 3.7278 26.0370 128.17 1 0.02908 0.20314 2 4 1.3910 7.6111 142.20 2 0.00978 0.05352 3 4 1.3835 12.0000 142.20 3 0.00973 0.08439 4 4 1.0671 11.9630 154.21 4 0.00692 0.07758 5 4 0.0000 13.5741 156.23 5 0.00000 0.08689 6 4 0.9915 0.0000 166.22 6 0.00597 0.00000 7 4 0.0000 9.4630 170.25 7 0.00000 0.05558 8 4 6.3817 11.2222 178.23 8 0.03581 0.06296 9 4 7.0939 10.9074 202.26 10 0.03507 0.05393 10 4 3.6973 6.2778 202.26 11 0.01828 0.03104 11 4 3.0205 4.9074 228.29 12 0.01323 0.02150 12 4 1.2794 0.0000 228.29 13 0.00560 0.00000 13 4 2.5340 4.4444 252.32 14 0.01004 0.01761 14 4 24.7897 20.7963 252.32 15 0.09825 0.08242 15 4 4.4511 10.4074 252.32 16 0.01764 0.04125 16 5 4.2986 8.3673 128.17 1 0.03354 0.06528 17 5 2.2823 3.3061 142.20 2 0.01605 0.02325 18 5 2.0964 4.5918 142.20 3 0.01474 0.03229 19 5 1.5249 2.8980 154.21 4 0.00989 0.01879 20 5 0.0000 5.3061 156.23 5 0.00000 0.03396 21 5 0.0000 2.6735 166.22 6 0.00000 0.01608 22 5 1.4302 3.3469 170.25 7 0.00840 0.01966 23 5 9.4287 3.9184 178.23 8 0.05290 0.02198 24 5 1.1943 1.0204 192.26 9 0.00621 0.00531 25 5 2.7115 2.6735 202.26 10 0.01341 0.01322 26 5 2.7236 1.3265 202.26 11 0.01347 0.00656 27 5 1.6731 1.5306 228.29 12 0.00733 0.00670 28 5 48.6282 21.6735 252.32 15 0.19272 0.08590 29 5 1.2731 0.0000 252.32 16 0.00505 0.00000 30 6 3.6883 5.6421 128.17 1 0.02878 0.04402 31 6 2.2485 5.2421 142.20 2 0.01581 0.03686 32 6 2.0274 5.0105 142.20 3 0.01426 0.03524 33 6 1.9501 2.6632 154.21 4 0.01265 0.01727 34 6 3.1882 3.4526 156.23 5 0.02041 0.02210 35 6 2.4097 1.8526 166.22 6 0.01450 0.01115 36 6 3.9054 2.1579 170.25 7 0.02294 0.01267 37 6 10.3210 3.6000 178.23 8 0.05791 0.02020 38 6 2.1980 1.0526 192.26 9 0.01143 0.00548 39 6 5.8612 5.3158 202.26 10 0.02898 0.02628 40 6 5.0932 3.2842 202.26 11 0.02518 0.01624 41 6 5.2747 3.0947 228.29 12 0.02311 0.01356 42 6 2.5364 1.0737 228.29 13 0.01111 0.00470
2.22

 43 6 2.0751 0.8211 252.32 14 0.00822 0.00325 44 6 13.9559 5.9053 252.32 15 0.05531 0.02340 45 6 3.5580 1.9474 252.32 16 0.01410 0.00772 46 7 5.014 5.9123 128.17 1 0.03912 0.04613 47 7 3.454 2.6316 142.20 2 0.02429 0.01851 48 7 3.939 3.5789 142.20 3 0.02770 0.02517 49 7 2.312 2.4737 154.21 4 0.01500 0.01604 50 7 2.896 3.4386 156.23 5 0.01853 0.02201 51 7 1.600 1.3158 166.22 6 0.00962 0.00792 52 7 1.622 1.8772 170.25 7 0.00953 0.01103 53 7 9.197 2.3509 178.23 8 0.05160 0.01319 54 7 1.527 1.1228 192.26 9 0.00794 0.00584 55 7 5.646 3.1053 202.26 10 0.02791 0.01535 56 7 4.740 2.5789 202.26 11 0.02344 0.01275 57 7 3.972 3.0175 228.29 12 0.01740 0.01322 58 7 1.362 0.6140 228.29 13 0.00597 0.00269 59 7 2.357 0.7193 252.32 14 0.00934 0.00285 60 7 190.789 62.2456 252.32 15 0.75614 0.24669 61 7 3.386 1.4035 252.32 16 0.01342 0.00556 62 9 4.952 6.0877 128.17 1 0.03864 0.04750 63 9 2.112 6.2807 142.20 2 0.01485 0.04417 64 9 3.648 8.1579 142.20 3 0.02566 0.05737 65 9 1.782 3.5088 154.21 4 0.01156 0.02275 66 9 3.469 8.6491 156.23 5 0.02220 0.05536 67 9 1.057 2.2456 166.22 6 0.00636 0.01351 68 9 1.321 3.0000 170.25 7 0.00776 0.01762 69 9 7.274 4.6667 178.23 8 0.04081 0.02618 70 9 0.000 1.8070 192.26 9 0.00000 0.00940 71 9 2.744 2.7719 202.26 10 0.01356 0.01370 72 9 2.532 1.8947 202.26 11 0.01252 0.00937 73 9 2.018 1.2807 228.29 12 0.00884 0.00561 74 9 1.819 0.3333 228.29 13 0.00797 0.00146 75 9 1.052 0.6667 252.32 14 0.00417 0.00264 76 9 70.149 35.6667 252.32 15 0.27802 0.14135 77 9 2.052 1.1579 252.32 16 0.00813 0.00459 78 10 5.103 4.6900 128.17 1 0.03981 0.03659 79 10 1.794 1.3400 142.20 2 0.01261 0.00942 80 10 1.835 2.6300 142.20 3 0.01290 0.01850 81 10 1.360 1.6200 154.21 4 0.00882 0.01051 82 10 1.261 2.6500 156.23 5 0.00807 0.01696 83 10 0.971 0.9100 166.22 6 0.00584 0.00547 84 10 1.175 1.2500 170.25 7 0.00690 0.00734 85 10 1.916 1.6300 178.23 8 0.01075 0.00915 86 10 1.466 0.8700 192.26 9 0.00762 0.00453 87 10 2.581 1.4000 202.26 10 0.01276 0.00692 88 10 1.785 1.2300 202.26 11 0.00883 0.00608 89 10 1.412 1.2500 228.29 12 0.00618 0.00548 90 10 0.000 0.1800 228.29 13 0.00000 0.00079 91 10 1.042 0.4700 252.32 14 0.00413 0.00186
2.23

 92 10 113.346 52.9900 252.32 15 0.44922 0.21001 93 10 1.006 0.4900 252.32 16 0.00399 0.00194 94 11 3.610 7.2371 128.17 1 0.02817 0.05646 95 11 1.830 2.5155 142.20 2 0.01287 0.01769 96 11 1.981 4.7216 142.20 3 0.01393 0.03320 97 11 1.585 2.1031 154.21 4 0.01028 0.01364 98 11 1.673 4.2474 156.23 5 0.01071 0.02719 99 11 0.000 14.5464 166.22 6 0.00000 0.08751 100 11 0.858 1.7938 170.25 7 0.00504 0.01054 101 11 1.735 2.4639 178.23 8 0.00974 0.01382 102 11 0.855 1.3299 192.26 9 0.00445 0.00692 103 11 2.051 2.4433 202.26 10 0.01014 0.01208 104 11 1.421 0.0000 202.26 11 0.00702 0.00000 105 11 1.413 2.2474 228.29 12 0.00619 0.00984 106 11 0.613 0.0000 228.29 13 0.00268 0.00000 107 11 0.709 0.0000 252.32 14 0.00281 0.00000 108 11 129.307 48.6082 252.32 15 0.51247 0.19265 109 11 1.672 0.0000 252.32 16 0.00663 0.00000 110 12 3.986 11.4694 128.17 1 0.03110 0.08949 111 12 1.608 5.9796 142.20 2 0.01131 0.04205 112 12 2.121 7.1429 142.20 3 0.01491 0.05023 113 12 1.727 3.6531 154.21 4 0.01120 0.02369 114 12 1.493 10.5918 156.23 5 0.00956 0.06780 115 12 1.071 3.7959 166.22 6 0.00645 0.02284 116 12 1.052 8.8980 170.25 7 0.00618 0.05226 117 12 2.053 19.2245 178.23 8 0.01152 0.10786 118 12 1.043 5.6939 192.26 9 0.00542 0.02962 119 12 2.587 4.5102 202.26 10 0.01279 0.02230 120 12 2.167 3.9184 202.26 11 0.01071 0.01937 121 12 1.548 1.6122 228.29 12 0.00678 0.00706 122 12 0.000 2.4490 228.29 13 0.00000 0.01073 123 12 0.876 0.0000 252.32 14 0.00347 0.00000 124 12 54.336 26.2653 252.32 15 0.21535 0.10410 125 12 1.057 0.0000 252.32 16 0.00419 0.00000 126 13 5.511 3.9242 128.17 1 0.04300 0.03062 127 13 2.642 3.6212 142.20 2 0.01858 0.02547 128 13 2.956 5.4242 142.20 3 0.02079 0.03815 129 13 2.530 1.4242 154.21 4 0.01641 0.00924 130 13 1.658 5.0303 156.23 5 0.01061 0.03220 131 13 0.805 0.8333 166.22 6 0.00484 0.00501 132 13 1.055 2.3788 170.25 7 0.00619 0.01397 133 13 1.927 1.7727 178.23 8 0.01081 0.00995 134 13 1.420 0.7121 192.26 9 0.00739 0.00370 135 13 3.165 2.0455 202.26 10 0.01565 0.01011 136 13 2.4696 1.8182 202.26 11 0.01221 0.00899 137 13 2.6575 1.1364 228.29 12 0.01164 0.00498 138 13 1.1395 0.4848 228.29 14 0.00499 0.00212 139 13 1.0265 0.5606 252.32 14 0.00407 0.00222 140 13 92.5926 37.9545 252.32 15 0.36696 0.15042
2.24

 141 13 1.4223 1.3788 252.32 16 0.00564 0.00546 142 14 4.0636 5.2432 128.17 1 0.03170 0.04091 143 14 2.2940 5.2793 142.20 2 0.01613 0.03713 144 14 2.2365 6.4685 142.20 3 0.01573 0.04549 145 14 1.3810 2.5676 154.21 4 0.00896 0.01665 146 14 1.6780 5.8739 156.23 5 0.01074 0.03760 147 14 1.1280 1.2973 166.22 6 0.00679 0.00780 148 14 1.1299 2.3243 170.25 7 0.00664 0.01365 149 14 1.9380 3.2793 178.23 8 0.01087 0.01840 150 14 1.0663 1.1622 192.26 9 0.00555 0.00604 151 14 2.3709 1.9459 202.26 10 0.01172 0.00962 152 14 2.4181 1.6577 202.26 11 0.01196 0.00820 153 14 2.0901 1.1351 228.29 12 0.00916 0.00497 154 14 0.5499 0.0000 228.29 13 0.00241 0.00000 155 14 0.7160 0.0000 252.32 14 0.00284 0.00000 156 14 49.2031 15.3694 252.32 15 0.19500 0.06091 157 14 1.2293 0.6036 252.32 16 0.00487 0.00239 158 16 5.4776 7.3816 128.17 1 0.04274 0.05759 159 16 2.2395 6.6974 142.20 2 0.01575 0.04710 160 16 2.1533 6.0526 142.20 3 0.01514 0.04256 161 16 1.6495 2.7237 154.21 4 0.01070 0.01766 162 16 1.5549 7.5132 156.23 5 0.00995 0.04809 163 16 1.3456 1.4868 166.22 6 0.00810 0.00895 164 16 1.5136 4.1974 170.25 7 0.00889 0.02465 165 16 6.3437 4.5263 178.23 8 0.03559 0.02540 166 16 1.0846 1.1579 192.26 9 0.00564 0.00602 167 16 4.8371 4.2237 202.26 10 0.02392 0.02088 168 16 4.3327 3.8684 202.26 11 0.02142 0.01913 169 16 3.9894 1.7500 228.29 12 0.01748 0.00767 170 16 1.4460 1.8816 228.29 13 0.00633 0.00824 171 16 1.8866 0.0000 252.32 14 0.00748 0.00000 172 16 26.3365 14.3816 252.32 15 0.10438 0.05700 173 16 1.9889 2.1974 252.32 16 0.00788 0.00871 174 19 5.0755 3.4023 128.17 1 0.03960 0.02655 175 19 1.8034 3.0805 142.20 2 0.01268 0.02166 176 19 1.9948 4.1839 142.20 3 0.01403 0.02942 177 19 1.7565 2.4023 154.21 4 0.01139 0.01558 178 19 1.5249 5.5057 156.23 5 0.00976 0.03524 179 19 1.1007 1.2529 166.22 6 0.00662 0.00754

 180 19 1.1964 3.4253 170.25 7 0.00703 0.02012 
2.25

 OBS POOL LLCONC FLCONC MOLEWT CHEM LMML FMML

 181 19 5.2950 4.3793 178.23 8 0.02971 0.024571 182 19 1.1791 1.6897 192.26 9 0.00613 0.008788 183 19 3.5176 2.6322 202.26 10 0.01739 0.013014 184 19 3.1881 2.5747 202.26 11 0.01576 0.012730 185 19 1.9302 0.9425 228.29 12 0.00846 0.004129 186 19 1.0474 0.7816 228.29 13 0.00459 0.003424 187 19 1.1345 0.5172 252.32 14 0.00450 0.002050 188 19 41.6354 18.0115 252.32 15 0.16501 0.071384 189 19 1.2650 0.8161 252.32 16 0.00501 0.003234 190 22 4.2526 4.1146 128.17 1 0.03318 0.032103 191 22 2.0454 2.0208 142.20 2 0.01438 0.014211 192 22 2.1461 3.1354 142.20 3 0.01509 0.022049 193 22 1.2772 1.8646 154.21 4 0.00828 0.012091 194 22 1.2666 4.8958 156.23 5 0.00811 0.031337 195 22 1.3352 1.5521 166.22 6 0.00803 0.009338 196 22 0.9046 6.4375 170.25 7 0.00531 0.037812 197 22 3.9079 2.7708 178.23 8 0.02193 0.015546 198 22 1.1377 0.8542 192.26 9 0.00592 0.004443 199 22 3.1845 2.6979 202.26 10 0.01574 0.013339 200 22 2.5594 2.4896 202.26 11 0.01265 0.012309 201 22 2.3395 1.2188 228.29 12 0.01025 0.005339 202 22 0.8239 0.6458 228.29 13 0.00361 0.002829 203 22 0.9537 0.5625 252.32 14 0.00378 0.002229 204 22 25.0994 18.6667 252.32 15 0.09947 0.073980 205 22 1.5719 0.9583 252.32 16 0.00623 0.003798
2.26

Appendix 2.5. Ratios of laboratory to field (L:F) and field to laboratory (F:L) tissue concentrations. Ratios were normalized tissue concentrations. Lipid-normalized concentrations (μg/g lipid) are listed for laboratory-exposed (collected (FLCONC) oligochaetes. 

 OBS CHEMICAL CHEM # POOL LLCONC FLCONC 

 1 1,6,7-TRIMETHNAPH 7 5 1.43018 3.3469  2 1,6,7-TRIMETHNAPH 7 6 3.90541 2.1579  3 1,6,7-TRIMETHNAPH 7 7 1.62179 1.8772  4 1,6,7-TRIMETHNAPH 7 9 1.32051 3.0000  5 1,6,7-TRIMETHNAPH 7 10 1.17534 1.2500  6 1,6,7-TRIMETHNAPH 7 11 0.85849 1.7938  7 1,6,7-TRIMETHNAPH 7 12 1.05160 8.8980  8 1,6,7-TRIMETHNAPH 7 13 1.05454 2.3788  9 1,6,7-TRIMETHNAPH 7 14 1.12992 2.3243  10 1,6,7-TRIMETHNAPH 7 16 1.51357 4.1974  11 1,6,7-TRIMETHNAPH 7 19 1.19635 3.4253  12 1,6,7-TRIMETHNAPH 7 22 0.90463 6.4375  13 1-METHYLNAPH 2 4 1.39105 7.6111  14 1-METHYLNAPH 2 5 2.28229 3.3061  15 1-METHYLNAPH 2 6 2.24855 5.2421  16 1-METHYLNAPH 2 7 3.45378 2.6316  17 1-METHYLNAPH 2 9 2.11157 6.2807  18 1-METHYLNAPH 2 10 1.79360 1.3400  19 1-METHYLNAPH 2 11 1.82974 2.5155  20 1-METHYLNAPH 2 12 1.60816 5.9796  21 1-METHYLNAPH 2 13 2.64189 3.6212  22 1-METHYLNAPH 2 14 2.29404 5.2793  23 1-METHYLNAPH 2 16 2.23951 6.6974  24 1-METHYLNAPH 2 19 1.80344 3.0805 
2.27

 25 1-METHYLNAPH 2 22 2.04537 2.0208  26 1-METHYLPHEN 9 5 1.19430 1.0204  27 1-METHYLPHEN 9 6 2.19802 1.0526  28 1-METHYLPHEN 9 7 1.52692 1.1228  29 1-METHYLPHEN 9 10 1.46592 0.8700  30 1-METHYLPHEN 9 11 0.85487 1.3299  31 1-METHYLPHEN 9 12 1.04266 5.6939  32 1-METHYLPHEN 9 13 1.42040 0.7121  33 1-METHYLPHEN 9 14 1.06633 1.1622  34 1-METHYLPHEN 9 16 1.08460 1.1579  35 1-METHYLPHEN 9 19 1.17914 1.6897  36 1-METHYLPHEN 9 22 1.13769 0.8542  37 2,6-DIMETHNAPH 5 6 3.18818 3.4526  38 2,6-DIMETHNAPH 5 7 2.89561 3.4386  39 2,6-DIMETHNAPH 5 9 3.46864 8.6491  40 2,6-DIMETHNAPH 5 10 1.26107 2.6500  41 2,6-DIMETHNAPH 5 11 1.67319 4.2474  42 2,6-DIMETHNAPH 5 12 1.49311 10.5918  43 2,6-DIMETHNAPH 5 13 1.65836 5.0303  44 2,6-DIMETHNAPH 5 14 1.67797 5.8739 

 45 2,6-DIMETHNAPH 5 16 1.55492 7.5132 

 46 2,6-DIMETHNAPH 5 19 1.52495 5.5057  47 2,6-DIMETHNAPH 5 22 1.26658 4.8958  48 2-METHYLNAPH 3 4 1.38352 12.0000  49 2-METHYLNAPH 3 5 2.09642 4.5918  50 2-METHYLNAPH 3 6 2.02741 5.0105  51 2-METHYLNAPH 3 7 3.93896 3.5789  52 2-METHYLNAPH 3 9 3.64825 8.1579  53 2-METHYLNAPH 3 10 1.83465 2.6300  54 2-METHYLNAPH 3 11 1.98082 4.7216 
2.28

 55 2-METHYLNAPH 3 12 2.12090 7.1429  56 2-METHYLNAPH 3 13 2.95584 5.4242  57 2-METHYLNAPH 3 14 2.23649 6.4685  58 2-METHYLNAPH 3 16 2.15334 6.0526  59 2-METHYLNAPH 3 19 1.99482 4.1839  60 2-METHYLNAPH 3 22 2.14610 3.1354  61 ACENAPHTHENE 10 0.80778 0.3800  62 ACENAPHTHENE 12 1.00739 1.3265  63 ACENAPHTHENE 13 0.69608 0.5758  64 ACENAPHTHENE 14 0.90012 0.6667  65 ACENAPHTHENE 16 1.13831 1.0132  66 BENaANTHRACENE 13 6 2.53638 1.0737  67 BENaANTHRACENE 13 7 1.36242 0.6140  68 BENaANTHRACENE 13 9 1.81888 0.3333  69 BENaANTHRACENE 13 13 1.13952 0.4848  70 BENaANTHRACENE 13 16 1.44604 1.8816  71 BENaANTHRACENE 13 19 1.04739 0.7816  72 BENaANTHRACENE 13 22 0.82395 0.6458  73 BENbkFLUORAN 14 4 5.06801 8.8889  74 BENbkFLUORAN 14 6 4.15012 1.6421  75 BENbkFLUORAN 14 7 4.71436 1.4386  76 BENbkFLUORAN 14 9 2.10390 1.3333  77 BENbkFLUORAN 14 10 2.08379 0.9400  78 BENbkFLUORAN 14 13 2.05292 1.1212  79 BENbkFLUORAN 14 19 2.26898 1.0345  80 BENbkFLUORAN 14 22 1.90736 1.1250  81 BENePYRENE 16 4 4.45106 10.4074  82 BENePYRENE 16 6 3.55795 1.9474  83 BENePYRENE 16 7 3.38590 1.4035  84 BENePYRENE 16 9 2.05173 1.1579 
2.29

 85 BENePYRENE 16 10 1.00561 0.4900  86 BENePYRENE 16 13 1.42232 1.3788  87 BENePYRENE 16 14 1.22928 0.6036  88 BENePYRENE 16 16 1.98890 2.1974  89 BENePYRENE 16 19 1.26503 0.8161  90 BENePYRENE 16 22 1.57189 0.9583  91 BIPHENYL 4 4 1.06709 11.9630  92 BIPHENYL 4 5 1.52488 2.8980  93 BIPHENYL 4 6 1.95013 2.6632  94 BIPHENYL 4 7 2.31239 2.4737  95 BIPHENYL 4 9 1.78193 3.5088  96 BIPHENYL 4 10 1.35955 1.6200  97 BIPHENYL 4 11 1.58544 2.1031  98 BIPHENYL 4 12 1.72667 3.6531  99 BIPHENYL 4 13 2.52996 1.4242  100 BIPHENYL 4 14 1.38104 2.5676  101 BIPHENYL 4 16 1.64953 2.7237  102 BIPHENYL 4 19 1.75647 2.4023  103 BIPHENYL 4 22 1.27716 1.8646  104 C1-NAPHTHALENES 4 2.77457 19.6111  105 C1-NAPHTHALENES 5 3.58325 7.8980  106 C1-NAPHTHALENES 6 4.28271 10.2421  107 C1-NAPHTHALENES 7 7.39486 6.2105  108 C1-NAPHTHALENES 9 5.75268 14.4386  109 C1-NAPHTHALENES 10 3.63483 3.9700  110 C1-NAPHTHALENES 11 3.81527 7.2371  111 C1-NAPHTHALENES 12 3.72231 13.1224  112 C1-NAPHTHALENES 13 5.59283 9.0455  113 C1-NAPHTHALENES 14 4.52395 11.7477  114 C1-NAPHTHALENES 16 4.39785 12.7500 
2.30

 115 C1-NAPHTHALENES 19 3.80030 7.2644  116 C1-NAPHTHALENES 22 4.18440 5.1563  117 CHRYSENE 12 4 3.02049 4.9074  118 CHRYSENE 12 5 1.67312 1.5306  119 CHRYSENE 12 6 5.27471 3.0947  120 CHRYSENE 12 7 3.97181 3.0175  121 CHRYSENE 12 9 2.01778 1.2807  122 CHRYSENE 12 10 1.41153 1.2500  123 CHRYSENE 12 11 1.41262 2.2474  124 CHRYSENE 12 12 1.54802 1.6122  125 CHRYSENE 12 13 2.65747 1.1364  126 CHRYSENE 12 14 2.09014 1.1351  127 CHRYSENE 12 16 3.98941 1.7500  128 CHRYSENE 12 19 1.93023 0.9425  129 CHRYSENE 12 22 2.33951 1.2188  130 FLUORANTHENE 11 4 3.69733 6.2778  131 FLUORANTHENE 11 5 2.72360 1.3265  132 FLUORANTHENE 11 6 5.09321 3.2842  133 FLUORANTHENE 11 7 4.74009 2.5789  134 FLUORANTHENE 11 9 2.53183 1.8947  135 FLUORANTHENE 11 10 1.78517 1.2300  136 FLUORANTHENE 11 12 2.167 3.9184  137 FLUORANTHENE 11 13 2.470 1.8182  138 FLUORANTHENE 11 14 2.418 1.6577  139 FLUORANTHENE 11 16 4.333 3.8684  140 FLUORANTHENE 11 19 3.188 2.5747  141 FLUORANTHENE 11 22 2.559 2.4896  142 FLUORENE 6 6 2.410 1.8526  143 FLUORENE 6 7 1.600 1.3158  144 FLUORENE 6 9 1.057 2.2456 
2.31

 145 FLUORENE 6 10 0.971 0.9100  146 FLUORENE 6 12 1.071 3.7959  147 FLUORENE 6 13 0.805 0.8333  148 FLUORENE 6 14 1.128 1.2973  149 FLUORENE 6 16 1.346 1.4868  150 FLUORENE 6 19 1.101 1.2529  151 FLUORENE 6 22 1.335 1.5521  152 NAPHTHALENE 1 4 3.728 26.0370  153 NAPHTHALENE 1 5 4.299 8.3673  154 NAPHTHALENE 1 6 3.688 5.6421  155 NAPHTHALENE 1 7 5.014 5.9123  156 NAPHTHALENE 1 9 4.952 6.0877  157 NAPHTHALENE 1 10 5.103 4.6900  158 NAPHTHALENE 1 11 3.610 7.2371  159 NAPHTHALENE 1 12 3.986 11.4694  160 NAPHTHALENE 1 13 5.511 3.9242  161 NAPHTHALENE 1 14 4.064 5.2432  162 NAPHTHALENE 1 16 5.478 7.3816  163 NAPHTHALENE 1 19 5.075 3.4023  164 NAPHTHALENE 1 22 4.253 4.1146  165 PERYLENE 15 4 24.790 20.7963  166 PERYLENE 15 5 48.628 21.6735  167 PERYLENE 15 6 13.956 5.9053  168 PERYLENE 15 7 190.789 62.2456  169 PERYLENE 15 9 70.149 35.6667  170 PERYLENE 15 10 113.346 52.9900  171 PERYLENE 15 11 129.307 48.6082  172 PERYLENE 15 12 54.336 26.2653  173 PERYLENE 15 13 92.593 37.9545  174 PERYLENE 15 14 49.203 15.3694 
2.32

 175 PERYLENE 15 16 26.337 14.3816  176 PERYLENE 15 19 41.635 18.0115  177 PERYLENE 15 22 25.099 18.6667  178 PHENANTHRENE 8 4 6.382 11.2222  179 PHENANTHRENE 8 5 9.429 3.9184  180 PHENANTHRENE 8 6 10.321 3.6000  181 PHENANTHRENE 8 7 9.197 2.351  182 PHENANTHRENE 8 9 7.274 4.667  183 PHENANTHRENE 8 10 1.916 1.630  184 PHENANTHRENE 8 11 1.735 2.464  185 PHENANTHRENE 8 12 2.053 19.224  186 PHENANTHRENE 8 13 1.927 1.773  187 PHENANTHRENE 8 14 1.938 3.279  188 PHENANTHRENE 8 16 6.344 4.526  189 PHENANTHRENE 8 19 5.295 4.379  190 PHENANTHRENE 8 22 3.908 2.771  191 PYRENE 10 4 7.094 10.907  192 PYRENE 10 5 2.711 2.673  193 PYRENE 10 6 5.861 5.316  194 PYRENE 10 7 5.646 3.105  195 PYRENE 10 9 2.744 2.772  196 PYRENE 10 10 2.581 1.400  197 PYRENE 10 11 2.051 2.443  198 PYRENE 10 12 2.587 4.510  199 PYRENE 10 13 3.165 2.045  200 PYRENE 10 14 2.371 1.946  201 PYRENE 10 16 4.837 4.224  202 PYRENE 10 19 3.518 2.632  203 PYRENE 10 22 3.185 2.698  204 TOTAL PCBs 4 133.321 149.278 
2.33

 205 TOTAL PCBs 5 26.355 66.061  206 TOTAL PCBs 6 28.644 13.768  207 TOTAL PCBs 7 14.275 99.316  208 TOTAL PCBs 9 15.253 17.000  209 TOTAL PCBs 10 23.962 14.350  210 TOTAL PCBs 11 39.490 19.979  211 TOTAL PCBs 12 25.261 21.204  212 TOTAL PCBs 13 10.182 8.197  213 TOTAL PCBs 14 14.400 27.045  214 TOTAL PCBs 16 58.854 25.026  215 TOTAL PCBs 19 50.689 6.310  216 TOTAL PCBs 22 25.622 5.813 
3.1

Chapter 3: Assessing Sediment Toxicity from Upper Mississippi River

Navigational Pools Using a Benthic Invertebrate Community Evaluation

and the Sediment Quality Triad Approach

Canfield, T.J., Brunson, E.L., Dwyer, F.J., Ingersoll, C.G., and

Kemble, N.E.

Introduction

The Mississippi River is the central catchment for a majority of the

water runoff between the west side of the Appalachian mountain range

to the east side of the Rocky mountain range. This makes the

Mississippi River system the largest in the United States, the third

largest drainage worldwide, and the seventh largest average discharge

worldwide (Van der Leeden, Troise and Todd 1990). The river receives

inputs from municipal, agricultural, and industrial sources. Previous

studies have examined the concentrations of organic and inorganic

contaminants in the sediments from select pools in the Upper

Mississippi River (Wiebe 1927; Bailey and Rada 1984; Weiner et al.

1984; Rada et al. 1990). Recently, some studies have reported a

decline in the levels of contaminants in the sediments of the Upper

Mississippi River (Rada et al. 1990).

The Upper Mississippi River (UMR), that part of the river north of

the confluence with the Ohio River at Cairo, IL, is divided into a

series of large runs and pools by 26 locks and dams constructed for

navigational purposes (Rada et al. 1990). This lock and dam system,

which runs from Minneapolis, MN to St. Louis, MO, provides areas for

deposition of large quantities of fine grained sediments during

normal and low flows (Nielson et al. 1984). Contaminants are often

associated with fine-grained sediments and settle along with these

sediments.(Forstner and Wittmann 1979, Hassett et al. 1980) 

Sediments often serve as a sink for an array of organic and inorganic

contaminants when the water to sediment gradient is high, and these

sediments can act as a source of contamination when the water to

sediment gradient is low (Shimp et al. 1971; Oschwald 1972; Medine and

McCutcheon 1989). 

Benthic macroinvertebrates inhabiting the sediments are presumably

exposed continuously to any contaminants contained in the sediments. 

Benthic macroinvertebrate abundance, community structure, and

ecological function have long been used to characterize water quality

in freshwater ecosystems (Davis and Lathrop 1992). Numerous studies
3.2

have documented potential changes in benthic invertebrate community

structure associated with the impacts of contaminants (Cook and

Johnson 1974; Rosenberg and Wiens 1976; Hilsenhoff 1982, 1987;

Waterhouse and Farrell 1985; Clements et al. 1992). Most studies in

lotic environments have examined the responses of benthic

macroinvertebrate communities in riffle areas due to ease of

collection and observed higher taxa richness. However, only a limited

number of assessments have been conducted in depositional softsediments (Canfield et al. 1996). 

The spatial and temporal distribution of resident organisms may

reflect the degree to which chemicals in the sediments are

bioavailable and toxic. Field surveys of invertebrates can provide an

important component of biological assessments of toxicity associated

with contaminated sediments for several reasons: (1)

macroinvertebrates are abundant, relatively sedentary, easy to

collect, and ubiquitous across a broad array of sediment types; (2)

many indigenous benthic organisms complete all or most of their life

cycles in the aquatic environment and may serve as continuous monitors

of sediment quality; and (3) results of an assessment of indigenous

populations may be useful for quantifying resource damage (Cook 1976,

Pratt and Coler 1976, Davis and Lathrop 1989).

The United States Geological Survey (USGS) has been monitoring the

UMR since 1987 to document the fate, transport, and distribution of

contaminated sediments (Moody and Meade 1995). Concern with regard to

the fate of contaminated sediments in the UMR arose after the flood of

1993, because of the potential for re-exposure of deeply buried,

potentially highly contaminated sediments. Further, the flood

inundated numerous riparian areas known to contain both diffuse and

concentrated (i.e. fuel tanks, warehouses) sources of contaminants. 

This study was designed to evaluate the current status of sediments in

the UMR and is one chapter in a series designed to assess the extent

of sediment contamination in the navigational pools of the river. The

overall study consisted of the following components: (1) monitoring

concentrations of contaminants in the Mississippi River sediments

(Moody et al. 1996); (2) toxicity testing with whole-sediments

collected from the river (Chapter 1); (3) bioaccumulation tests with

whole-sediments collected from the river (Chapter 2); and (4) 

analysis of benthic invertebrate community structure. The objective

of this portion of the study was three-fold: (1) describe

distributions and abundances of benthic invertebrates in softsediments from selected locations in pools of the UMR; (2) evaluate
3.3

impacts of contaminants associated with these sediments using measures

of benthic invertebrate community structure; and (3) evaluate the

concordance of benthic invertebrate assessments to sediment toxicity

and sediment chemistry using the sediment quality triad approach. 

Materials and Methods

Sampling Locations

Stations were selected for assessment of sediment toxicity, sediment

chemistry and benthic macroinvertebrate communities based on

historical chemistry data (Moody et al. 1996) and the availability of

soft sediments (Chapter 1). Upper Mississippi River pools were

sampled from June 11 to July 5, 1994. Stations were located in 23 of

26 pools in the UMR from pool 1 near Hastings, MN to pool 26 near St.

Louis, MO (Figure 1.1, Chapter 1). A complete description of the

sampling locations in each pool is described in Kemble et al. (Chapter

1) and bioaccumulation data is contained in Brunson et al. (Chapter

2).

Sediment Collection, Handling, and Storage 

Locations of stations for field sampling were determined with a Global

Positioning System. A stainless steel standard Ponar grab (23 x 23

cm, 529 cm area) was used to collect bulk sediments from about the 2

upper 6 to 10 cm of the sediment for chemistry analyses, laboratory

toxicity assessments and benthic invertebrates assessments at one

station per pool (Chapter 1). Each sample was a composite of 35 to 80

L of sediment/station (identified as C samples in Chapter 1). 

Sediments were placed in a 120-L high density polyethylene drum and

homogenized with a hand-held power drill and a stainless steel auger.

A 2.5 liter subsample of sediment for evaluations of benthos was

obtained taken from the composite C sample before subsamples were

obtained for chemistry and laboratory analyses (Chapter 1). To

isolate the benthos, these 2.5 liter subsamples were sieved through an

ASTM No.30 (533 μm) and an ASTM No. 60 (250 μm) bucket connected in

series using screened river water for rinsing. Material containing

benthos retained by the sieves was combined and transferred into 1L

high density polyethylene jars, preserved with 10% buffered formalin,

and transported to the laboratory. Subsamples for use in toxicity and

bioaccumulation testing (10L), for chemical characterization (250 ml
3.4

for metals, 250 ml for organics), and for physical characterization

(250 ml) were taken and stored in high-density polyethylene containers

or amber glass I-CHEM bottles (chemical characterizations only). All

samples were stored at 4 C in the dark (Chapter 1). o

Taxonomic Identification 

The preserved samples of benthos were placed in a sieve (250 μm) and

rinsed thoroughly with tap water in the laboratory to remove formalin

and excess silt or mud before sorting. The samples were drained of

excess water, returned to the original jars, filled with 95% ethanol

and allowed to soak for at least 24 h to facilitate extraction of

volatile compounds. Aliquots of the sample were sequentially removed

from the jar to sort benthic invertebrates until the entire sample had

been sorted. 

A binocular dissecting microscope (4x to 12x power) was used to

sort and pick the entire sample. Invertebrates were initially sorted

and enumerated into the following orders or families: Oligochaeta,

Chironomidae, Bivalvia, Gastropoda, Ephemeroptera, Odonata,

Plecoptera, Hemiptera, Megaloptera, Trichoptera, Coleoptera, Diptera,

Hirudinea, and Amphipoda. Taxa were identified to the lowest

practical level using appropriate taxonomic keys (Wiederholm 1983;

Merritt and Cummins 1984; Pennak 1989; Thorp and Covich 1991). The

following benthic macroinvertebrate metrics were calculated: 

macroinvertebrate abundance (number/m ), species composition, and taxa 2

richness (Appendix 3.1, 3.2, 3.3). All taxa were either identified

or verified by personnel at the Aquatic Resources Center in Franklin,

TN. 

Chironomid larvae (midge) were examined for deformities in

mouthpart structures. These deformities, which included various types

of asymmetry, missing teeth, extra teeth, fusion among various teeth,

and labial separation, have been described by several investigators

(Saether 1970; Hamilton and Saether 1971; Hare and Carter 1976;

Warwick et al. 1987; Warwick 1989). Individual midge were mounted on

slides and their mouthparts were examined for deformities in the

mentum and ligula (Tanypodinae only). Occurrence of deformities was

expressed as a proportion of the total number of midges at each

station.

Physical and Chemical Characterizations of Sediment
3.5

Sediment physical characteristics included the following: (1) sediment

particle size, (2) total organic carbon, (3) inorganic carbon and (4)

percent water. Sediment chemical parameters included the following: 

(1) chlorinated pesticides, (2) polychlorinated biphenyls (PCB), (3)

select aliphatic and polynuclear aromatic hydrocarbons (PAH), (4)

simultaneously extracted metals (SEM), (5) acid volatile sulfide

(AVS), and (6) total metals. See Chapter 1 for additional information

on chemical and physical characteristics of the sediments. 

Statistical Analyses

Statistical analysis was performed with the Statistical Analysis

System (SAS, Statistical Analysis System 1994). Comparisons between

benthic invertebrate abundance and physical and chemical data were

made with a Spearman Rank correlation and multivariate regression. If

not reported, statements of statistical significance indicate p < 0.05.

Sediment Quality Triad Assessments

The sediment quality triad (Triad) approach was used as an effects

based approach to integrate data from chemical and physical analyses

(e.g., PAH’s, metals, grain size), sediment laboratory toxicity

exposures (e.g., Hyalella azteca survival and growth) and benthic

community structure (e.g. biotic index, taxa richness, midge mouthpart deformities) in order to evaluate the level of concordance

between these three measures and the degree of contaminant-induced

degradation in aquatic communities in soft-sediment depositional areas

(Chapman et al. 1992). Toxicity, benthos, and chemistry data were

scored using procedures developed by Kreis (1988) and data were

plotted using procedures described by Canfield et al. (1994, 1996). 

Values for each individual variable for all samples were scaled

proportionally between 1 and 100 (e.g., 1 is indicative of the lowest

concentration or least impacted, and 100 is the greatest concentration

or most impacted). Scaling data retains proportional differences

between measurements and results in an identical range for all

variables. Typically, more than one variable is determined for a

particular Triad component (e.g. Hyalella azteca survival and growth). 

In these instances Kreis (1988) recommends: (1) scaling each

individual variable among samples, (2) summing the scaled values for

each variable, and then (3) re-scaling the sums for all samples. 

This results in scaled scores (e.g., toxicity, benthos, or chemistry)
3.6

between 1 and 100 for each Triad component which can be compared

graphically or in tabular form. 

The high and low values used to establish scores for each of the

sets of information for benthos, chemistry, and laboratory toxicity

were previously reported in Canfield et al. (1996). In order to

evaluate the extent of contamination of the UMR in the context of

other areas of concern in North America, we used data from three Great

Lakes Areas of Concern (Canfield et al. 1996) and data from a study of

the upper Clark Fork River, including Milltown Reservoir, in Montana

(Canfield et al. 1994). Inclusion of these data sets provided a

larger number of stations with a broad range in levels of

contamination that could be used in the analyses of the relative

responses of benthic communities in select sampling locations in the

UMR and evaluate the relative contamination of sediments in the UMR

sediments when compared to other areas. Six benthic invertebrate

indices were used to evaluate the extent of sediment contamination in

the UMR system: (1) total taxa richness, (2) chironomid genera

richness, (3) chironomid mouthpart deformities, (4) chironomid biotic

index, (5) chironomid/oligochaete ratio, and (6) oligochaete biotic

index. The Hilsenhoff index of Biotic Integrity was used to calculate

the biotic indices for both the midges and oligochaetes. Species

sensitivity within a genera was obtained primarily from those assigned

by Hilsenhoff (1982, 1987) and secondarily by Lenat (1993). 

To evaluate the chemistry portion of the Triad, we used EffectRange Median (ERM) concentrations calculated by Ingersoll et al.

(1996). An ERM is defined as the concentration of a chemical in

sediment above which effects are frequently or always observed (Long

et al. 1995). We used seven ERM values which correctly classify

laboratory toxicity >70% of the time in Hyalella azteca 28-d tests

(Ingersoll et al. 1996). These seven ERMs would more closely identify

cause and effect toxicity rather than correlative toxicity (Canfield

et al. 1996). These ERMs included: cadmium, nickel, lead, zinc,

chrysene, benzo(a)pyrene, and benzo(g,h,i)perylene. 

To evaluate the laboratory toxicity portion of the Triad we used

amphipod (Hyalella azteca) 28-d growth and survival to score

laboratory toxicity (Chapter 1). Amphipods are sensitive to

contaminated sediments and frequently exhibit reduced survival and

growth following exposures to contaminated sediments (Burton et al.

1996, Ingersoll et al. 1996). Each sample was designated as toxic

when either survival or growth of Hyalella were significantly reduced

relative to the control or reference sediment (Chapter 1; Kemble et
3.7

al. 1994; Ingersoll et al. 1996; USEPA 1993 ). Associations between

benthic indices or laboratory toxicity tests and sediment chemistry 

were evaluated by plotting the scores of either the benthic indices or

laboratory toxicity data against the sum of the ERM quotient (SERM-Q: 

ERM-Q=concentration of a chemical in sediment sample / ERM for that

chemical) for all seven chemicals in a sample. This approach is

similar to a toxic unit approach. 

Well defined guidelines have not been developed for distinguishing

impacts of contaminant effects on benthos found in soft sediments in

either lakes, streams, or rivers. Canfield et al. (1996) incorporated

data plots (partitioned by using quadrants defined by no effect

concentration data for plotting) and frequency analysis to identify

the distribution of the data points to identify relations between

sediment chemistry, laboratory toxicity and benthic invertebrate

distributions. This quadrant frequency analysis (essentially a

frequency analysis to identify correct classification and Type I and

Type II error) was conducted in order to evaluate which benthic

indices were most sensitive to elevated contaminant concentrations. 

In this analysis, scores for benthic indices are plotted against

scores for chemical contamination in sediments. Quadrants were then

defined which identified one of four possible conditions: (1) low

chemical concentration and benthos not adversely impacted, (2)

elevated chemical concentration and benthos adversely impacted, (3)

low chemical concentration and benthos adversely impacted (Type I

error, false positive), and (4) elevated chemical concentration and

benthos not adversely impacted (Type II error, false negative). 

Various combinations of benthic indices were evaluated by adding the

individual scores and re-scoring. These analyses were conducted for

all possible combinations of the six scored benthic indices listed

above. 

Sediment toxicity studies were conducted on sediments from all of

the UMR pools (except pools 3 and 17, Chapter 1). The results of the

tests on the UMR sediments were combined with data from 19 Great Lakes

sediment samples (Ingersoll et al. 1996) and 13 Clark Fork

River/Milltown Reservoir samples (Kemble et al. 1994) in order to

evaluate the toxicity of the UMR sediments in the context of other

samples previously evaluated. Based on previous plots of toxicity

scores (Canfield et al. 1996), the vertical quadrant line above which

no non-toxic samples were observed and above which chemical

contamination was considered toxic was sum of the ERM quotient of 39

(Figure 3.1). The horizontal quadrant line depicting laboratory
3.8

toxicity was a score of 30, which corresponded to the greatest

laboratory score above which no non-toxic samples were observed. 

This selection procedure for establishing quadrant lines may be less

environmentally protective since some of the samples that had a sum of

the ERM quotient score less than 39 were toxic to Hyalella azteca in

the laboratory studies (Chapter 1; Kemble et al. 1994; Ingersoll et

al. 1996). 

Scores for each of the benthic indices and all combinations of

scores were plotted against the sum of the ERM quotient. The position

of quadrant lines for benthic indices were determined in 3 steps: (1)

plotting the data, (2) drawing the vertical quadrant line at 39 for

the Sum of the ERM quotient, (3) by evaluating the distribution of

the data and selecting a benthic score (horizontal quadrant line)

which maximized the number of points in quadrants which would be

considered "correctly classified" and minimized the number of samples

with "Type I, false positive" and "Type II, false negative" error

results. 

Results and Discussion

Benthic Invertebrate Assessments

Abundance:

Benthic invertebrates from the UMR exhibited a wide range of

abundance values. Benthic invertebrate abundance (number/m ) in 2

samples ranged from 250/m in sample 1C to a maximum of 22,389/m in 2 2

sample 19C (Table 3.1). Total abundance values were less than 8,000/m 2

in 21 of 24 samples with the remaining 3 samples having abundance

values two-fold greater than any of the other samples. Oligochaetes

were numerically dominant in 12 of 24 samples. Midge comprised the

majority of the community in 8 of 24 samples with the bivalves (2),

mayflies (1) and nematodes (1) comprising the majority of the

community in 4 of 24 samples (Table 3.1; Appendix 3.1, 3.2, 3.3).

Oligochaete abundance ranged from 63/m in sample 5C to 12,111/m in 2 2

sample 19C (Table 3.1). Across the pools there were order of

magnitude differences in abundance values. In general, oligochaete

abundance is lowest in samples from the upper pools (1 to 7) and

higher in the lower pools (Table 3.1). We expected these differences

to be explained by organic carbon and grain size although no

significant correlations were observed in the correlation analysis

(Table 3.2) evident with this data set. 
3.9

Chironomid abundance ranged from zero in samples from station 7C to

8,889/m in samples from station 15C (Table 3.1). Distribution of 2

midge was fairly even across this range. These values for chironomid

abundances were generally higher than those reported from contaminated

sediments in Milltown Reservoir/Clark Fork River (Canfield et al.

1994) or the Great Lakes (Canfield et al. 1996). 

Community Composition: 

Samples from the UMR had a fairly diverse benthic invertebrate

community (Table 3.1). Overall taxa richness was greater in samples

from the lower two-thirds of the river than in the upper 8 pools. 

Oligochaete abundance accounted for 5 to 90% of the community in all

samples. Combined oligochaete and midge abundance accounted for 8 to

100% of the total benthic invertebrate community in all samples, with

the remainder of the benthic community abundance coming primarily from

the Bivalvia and Ephemeroptera. 

The oligochaete community was comprised of 2 families, 5 genera and

9 species (Appendix 3.1). Samples from 20C and 11C had the highest

number of species, while samples from 1C, 4C and 5C each had only one

species. Except for 10C, the oligochaete community was made up

entirely of the family Tubificidae. Limnodrilus spp., generally

considered tolerant of organic and metal contamination (Kennedy 1965,

Brinkhurst et al. 1972, Burt et al. 1991), was the most common genera

occurring in samples from the UMR. 

The midge community was comprised of 4 subfamilies (Chironomini,

Tanipodinae, Tanytarsini, Orthocladinae) and 18 genera (Appendix 3.2). 

The sample from station 10C had the highest number of genera present

(8), while sample from station 7C had no genera present. Chironomus

spp. was the most abundant genera present in 17 of 24 samples, with

Procladius spp. the most abundant in 3 of the remaining samples. 

The Bivalvia (clams) and aquatic insects (excluding midge)

comprised a large part (>20%) of the community collected in 11 of 24

samples (Appendix 3.3). Bivalvia abundance ranged from zero in 7

samples to 16,722/m in sample 9C (Table 3.1). The Bivalvia were 2

present in 17 of 24 samples. Bivalvia abundance was greater than or

equal to 1,000/m in 5 of 24 samples. Bivalvia abundance of 16,722/m 2 2

in sample 9C is 1 to 2 orders of magnitude greater than all other

samples collected and comprises 77% of the overall community abundance

(Table 3.1) . The Bivalvia community was made up almost entirely of

Musculium transversum.
3.10

The Ephemeroptera (mayflies) were present in 16 of 24 samples. 

Ephemeroptera abundance ranged from absent in 8 samples to 3,278/m in 2

sample 19C (Table 3.1). Ephemeroptera abundance was greater than or

equal to 500/m in 6 of 24 samples, but were entirely absent in 8 of 24 2

samples. The Ephemeroptera community was comprised of 2 families, 2

genera and 3 species. The majority of the insect community

(chironomidae excluded) was comprised of Hexagenia sp. 

The estimated abundance values of benthic invertebrates collected

in this study are comparable with the values of invertebrates

collected in previous studies of the UMR (Eckblad et al. 1977; Butts

and Sparks 1982; Neuswanger, Taylor and Reynolds 1982; Eckblad 1986;

Jahn and Anderson 1986; Hornbach et al. 1989). Although there is some

variation among studies, abundances were within the same range

regardless of the study. In 1991, the Environmental Management

Program, Long Term Resource Monitoring Program (LTRMP) issued an

observation bulletin (LTRMP Observational Bulletin NO. 1, Eckblad

1991) which reported on the observed decline of macroinvertebrate

communities (primarily the fingernail clams) in the UMR. Data from

our study does not support the trends reported in the LTRMP report. 

Benthos abundances were above the low level warnings issued in the

LTRMP report. Differences in abundances may be due to natural spatial

or temporal variation in the invertebrate communities or conditions in

the river and sediments have changed between the time when the LTRMP

report was issued and when we conducted our study. 

Deformities in chironomids

The frequency of mouth part deformities in the midge community ranged

from a low of zero in samples from 11 stations to a maximum of 13% in

sample 20C (Figure 3.2). Deformities were present in 13 out of 24

samples in the UMR, although only 4 of 24 samples had deformities

which could be considered above the identified background levels of 3

to 4% (Dickman, Brindle and Benson 1992).

Different genera of midge exhibit different levels of

susceptibility or tolerance to contaminants (Hamilton and Saether

1971; Hare and Carter 1976; Warwick 1985, 1988; Wiederholm 1984). 

Some genera are quite intolerant and are eliminated from locations

with relatively low levels of contaminants, while other genera such as

Procladius spp., Chironomus spp. and Cryptochironomus spp. are more

tolerant and may persist in contaminated locations (Warwick 1985; Bode

1988). An association between increased contamination and the
3.11

presence of midge deformities has been observed by several

investigators (Hamilton and Saether 1971; Warwick 1985; Tennessen and

Gottfried 1983; Cushman 1984; Wiederholm 1984; Diggins and Stewart

1993). Deformities reported in these studies include thickening of

the exoskeleton, enlargement and darkening of the head capsule,

asymmetry in mouth parts, missing or fused lateral teeth, and antennal

deformities. None of the specimens examined in the present study

exhibited antennal deformities. Deformities observed in this study

occurred only in Procladius spp. and Chironomus spp. 

 The occurrence of midge deformities is reportedly less than 1% in

non-impacted or pre-industrialization communities (Wiederholm 1984;

Warwick et al. 1987). Background levels have been estimated at 3% to

4% (Dickman, Brindle and Benson 1992), and investigators have

suggested that frequency of deformities in the range of 5 to 25% or

greater are generally associated with moderate to severe contamination

(Wiederholm 1984; Warwick et al. 1987). Based on these criteria,

deformities in midge from the UMR indicate that sediments from only 4

samples would be classified as Amoderately contaminated@ (Figure 3.2). 

Deformities of midges in samples from the UMR were considerably lower

than those from contaminated sediments in studies from either the

Milltown Reservoir/Clark Fork River (Canfield et al. 1994) or the

Great Lakes (Canfield et al.1996). These data indicate that overall

the sediments in the UMR are uncontaminated relative to other

locations with documented occurrences of deformities in midges. 

Correlation Data

Spearman rank correlations were used to compare associations of

physical and chemical measures to benthic responses because of nonnormal distribution of data (Snedecor and Cochran 1982). Few

significant correlations were detected between benthic parameters and

either contaminants or abiotic factors evaluated (Table 3.2). 

Significant negative correlations were observed between total

Ephemeroptera abundance (r=-0.43) and total abundance (r=-0.54) with

percent sand. Conversely, significant positive correlations were

observed between clay and total numbers (r=0.59), bivalve abundance

(r=0.49), chironomid abundance (r=0.48, Ephemeroptera abundance

(r=0.47), number of chironomid genera (r=0.46), and number of

chironomid taxa (r=0.46). Positive correlations with clay and

negative correlations with sand imply that hydrological factors such

as current velocity may have been determinants of benthic
3.12

distributions. For example, clay dominated areas may support greater

benthic nymphs due to increased stability of physical habitat and

increased deposition of organic matter compared to sandy areas. 

However, abiotic causality is difficult to infer without additional,

manipulative studies. 

Significant correlations with measures of chemical contamination

were sporadic, observed comparing the measures of total abundance

(TOTAL) with zinc (Zn), number of oligochaete taxa (OTAXA) with

cadmium (Cd), the oligochaete biotic index (OLBI) with chrysene

(CHRYS) and cadmium, and total taxa richness (TXRICH) with chrysene,

benzo(a)pyrene (BAP), cadmium and nickel (Ni, Table 3.2). Although

these correlations were significant, they still explained no more than

35% of the total variability. Further, the number of positive and

negative correlations varied within a particular chemical. This makes

interpretation difficult, but may not be unexpected given that the

measured chemicals in almost all the sediments were extremely low

compared to sediments in other locations in the United States and the

relative weakness of non-parametric correlations as statistical tools. 

Sediment Quality Triad

Spearman rank correlations described above were used to make initial

comparisons between measures of the benthic invertebrate community to

measures of sediment chemistry or overlying water quality at the

sampling stations. While rank correlation analysis can be used to

demonstrate association among variables, this ranking of data

eliminates proportional relationships among variables by re-ranking

data to simple rank-order (e.g. 1,2,3,). Thus, this ranking can not

be used to adequately evaluate dose response relationships. For these

reasons, we evaluated benthic community and laboratory toxicity data

using a quadrant classification approach described below (see also

Canfield et al. 1996).

Results of toxicity and chemistry evaluations of UMR sediments

presented in Chapter 1 indicate these sediment samples were relatively

uncontaminated compared to other locations in the United States

(Kemble et al. 1994; Ingersoll et al 1996). We used the sediment

quality triad approach in order to evaluate how benthic communities

sampled from the UMR compared to other locations in the U.S. we

previously have evaluated (Canfield et al. 1994, 1996).

Scores for various benthic indices relating benthic alterations to

contaminant levels were previously identified using data sets from the
3.13

Great Lakes (Canfield et al. 1996) and the Clark Fork River in Montana

(Canfield et al. 1994). The scores were used to evaluate the scores

for samples from the current study. In the present study, benthos

samples were not classified as impacted or non-impacted a priori, but

rather samples were considered to be classified as Aincorrect@ only if

the scores were in the false positive or false negative error

quadrants as established in Canfield et al. (1996) (Figure 3.1). 

Four benthic indices (midge biotic index, midge richness, percent

midge deformities, and taxa richness) were previously found to provide

some degree of discrimination among samples from the Great Lakes with

differing degrees of contamination (Canfield et al. 1996). In the

present study, midge deformities (19%) had the smallest combined false

positive and false negative error rate relative to the sum of the ERM

quotient score (Table 3.3). Midge oligochaete ratio, midge biotic

index, midge taxa richness, and total taxa richness had a combined

false positive and false negative error rate of 34% to 35%. A

benthos score required to obtain this degree of discrimination was

always > 75%. 

In addition to assessments using single indices, the combined

scores of benthic indices were evaluated which provided the best

classification (smallest combined false positive and false negative

error). Quadrant classification using the sum of the ERM quotient

score of two to three combined benthic indices reduced the false

positive and false negative error rate to 19 to 24% , which is less

than all individually scored benthic indices except midge deformities

(Table 3.3). The various combinations of four to all six benthic

indices were not included in or discussed since the accuracy of

classification did not increase with combinations of more than three

benthic indices (Table 3.3). The combinations were restricted so that

the benthos score required to minimize false positive and false

negative error was a score no greater than 80 to 81. We were unable

to identify a combined score of less than 80 which minimized both

false positive and false negative error. The combined metric of midge

oligochaete ratio, midge taxa richness and total taxa richness

provided the combination which had the lowest false positive and false

negative error of 19% (Table 3.3). 

 Table 3.4 summarizes the classification of sediment samples based

on exceedances of scores for toxicity, chemistry, or benthos by

quadrant analyses as described in Canfield et al. (1996). Twenty-one

of 24 samples (88%) showed good agreement (i.e. all "minuses") among

all three measures of the Triad, which indicated that no contaminant
3.14

induced degradation was observed (Table 3.4). None of the samples

were scored with all pluses (i.e. evidence of contaminant induced

degradation). In one of the 24 (4%) samples laboratory toxicity and

sediment chemistry measures were in agreement, however the benthic

component was not in accordance. Similarly in one of the samples

sediment chemistry and benthos response are in agreement, yet toxicity

did not occur. High concordance among laboratory toxicity, chemistry,

and benthos is evidence that these sediment samples from the UMR were

relatively low in contamination or toxic effects compared to other

locations we have previously evaluated (Canfield et al. 1994, 1996).

Summary

Benthic invertebrate abundance values in sediment samples from the UMR

were comparable to values reported from relatively uncontaminated

sediments. The percent composition of the benthic invertebrate

community also indicates a relatively healthy community compared to

more contaminated locations. Oligochaetes and chironomids constituted

over 90% of the benthic invertebrate communities collected in 10 of 24

samples from the UMR, which is expected given the pre-dominance of

soft sediments. However, most of the UMR pools had a relatively high

diversity of representatives from orders other than the oligochaetes

and chironomids , which is different from observations from other

highly contaminated areas (Canfield et al. 1994, 1996). Further

benthic community indices were only weakly correlated with sediment

contaminants. 

The occurrence of midge deformities ranged from 0 to 13% in the UMR

pool samples, which were relatively low compared to those chironomids

from more highly contaminated sediments. Sediment Quality Triad

analyses classified a high percentage of the samples (88%) to be not

impacted. These data indicate that these sediment samples were

relatively uncontaminated.

Additional studies are needed to evaluate specific contaminant,

biotic, and abiotic factors controlling benthic communities in soft

sediments associated with backwater areas of both lotic and lentic

environments. Studies designed to evaluate benthic distributions in

relation to factors influencing variation on a local microhabitat

scale are necessary in order to reduce the variation in the relations

between sediment chemistry, habitat, and measures of benthic

invertebrate communities. These studies should greatly expand our

ability to evaluate environmental quality of ecosystems such as the
3.15

UMR. 

Acknowledgements The authors would like to thank Doug Hardesty, Shane

Reussler, Jeff Steevens and David Whites for all their field and

laboratory help on this project. We would like to thank James

Fairchild, David Mount and Parley Winger for constructive criticism

and review of this manuscript. We thank Pamela Haverland and Ellen

Eherhart for their statistical help in developing the benthic metrics. 
3.16

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0 10 20 30 40 50 60 70 8

0

102030405060708090

100

SERM -Q

HYALELLA 

GROWTH-SURVIVAL

AC

GL-MT UMR

TOXIC NON-TOXIC NON-TOXI

TOXIC

3.24

Fig. 3.1. Quadrant analysis with the Hyalella azteca toxicity score and the SERM-Q. Quadrants are labeled : A: fhigh toxicity, low chemistry), B: false negative (Type II error; low toxicity, high chemistry), C: non-impacted (low impacted (high toxicity, high chemistry). A dark square or diamond indicates toxic samples and an open square or samples in Hyalella azteca toxicity tests. A square (either dark or open) indicates data from samples collected in thReservoir/Clark Fork River, Montana, and a diamond (either dark or open) indicates data from samples collected in River.
1% , HISTORICAL BACKGROUND LEVELS

4%, CURRENT BACKGROUND LEVELS

1

S C

2

4

5

6

7

8

9

1 0

1 1

1 2

1 3

1 4

1 5

1 6

1 8

1 9

2 0

2 

05

1 01 52 0

POOL #

MIDGE DEFORMITIES (%)

4%, CURRENT BACKGROUND LEVELS

SC 4 6 8 10 12 14 16 19 2

5

1 5

3.25

Fig. 3.2. Comparison of chironomid deformities (%) for samples from the UMR samples. Solid line ( ) represe

background level of deformities (based on analysis of core samples of pre-industrialization sediments) and the dash4% current background levels from modern day studies of uncontaminated sediments.
3.26

Table 3.1. Percent contribution of each taxa to the overall total abundance estimates. Values for total abundance are actual number of taOligochaeta; CH- Chironomidae; BIVL- Bivalvia; EPHM- Ephemeroptera; ODON- Odonata; HEM- Hemiptera; TRI- Trichoptera; DAMP- Amphipoda; NEMA- Nematoda; HDRNID- Hydrachnida. Sample numbers designate the pools in the Upper Mississippi River The sample number SCC corresponds to the sampling site in the Saint Croix River. 

TAXA

SAMPLE OL CH BIVL EPHM ODON HEM TRI DPT HR AMP NEMA HSCC 32 63 2 0 0 0 0 0 0 0 2 1C 33 67 0 0 0 0 0 0 0 0 0 2C 43 36 0 20 0 0 0 0 0 0 0 4C 18 3 0 3 0 0 0 0 0 0 75 5C 11 33 22 22 0 0 11 0 0 0 0 6C 53 26 3 5 0 2 0 7 0 2 2 7C 12 0 62 22 0 0 1 0 0 1 1 8C 43 35 0 18 1 0 0 2 0 0 0 9C 5 3 77 9 0 0 1 0 2 4 0 10C 46 51 2 0 0 0 0 0 0 0 0 11C 63 29 1 5 0 0 1 0 0 0 1 12C 7 30 19 35 0 1 0 2 1 1 4 13C 39 27 14 6 0 0 0 0 10 5 0 14C 75 20 2 0 0 0 0 2 0 0 0 15C 53 44 1 1 0 0 0 1 0 0 1 16C 46 53 0 0 1 0 0 0 0 0 0 18C 28 69 3 0 0 0 0 0 0 0 1 19C 54 25 6 15 0 0 0 0 0 0 0 20C 90 6 1 2 1 0 1 1 0 0 0 21C 16 60 0 18 3 0 0 3 0 0 0 22C 67 30 2 0 0 0 2 0 0 0 0 
3.27

24C 13 75 2 6 0 0 0 4 0 0 0 25C 78 16 3 3 0 0 0 0 0 0 0 26C 55 45 0 0 0 0 0 0 0 0 0 
3.28

Table 3.2. Spearman rank correlation for whole sediment measured naphthalene (NAPH), chrysene (CHRYS), becadmium (Cd), nickel (Ni), lead (Pb), zinc (Zn), percent water (%H2O), percent sand (%SAND), percent silt (%SILtotal organic carbon (TOC) with oligochaete abundance (OL), chironomid midge abundance (CH), bivalve abundanabundance (EPHM), total abundance (TOTAL), number of oligochaete taxa (OTAXA), number of chironomid mioligochaete biotic index (OLBI), chironomid midge biotic index, ratio of oligochaetes to chironomid midge (MGOmouthpart deformities (MGDFRM), chironomid midge genera richness (MGRICH), and total taxa richness (TXRICare designated with an asterisk (*; P< 0.05).

METRIC NAPH CHRYS BAP Cd Ni Pb Zn %H2O %SAND %SILT OL 0.21 0.15 -0.09 -0.23 0.07 0.11 0.13 -0.07 -0.32 0.17 CH 0.04 0.03 -0.01 -0.06 -0.08 0.24 0.32 -0.17 -0.39 0.07 

BIVL 0.28 -0.04 -0.25 0.09 0.21 0.38 0.37 -0.24 -0.33 -0.17 EPHM 0.34 -0.21 -0.01 -0.10 0.16 0.0 0.04 -0.22 -0.43 0.09 

* 

TOTAL 0.39 0.08 0.0 0.10 0.26 0.39 0.46 -0.31 -0.54 0.09 

* *

OTAXA -0.20 -0.14 -0.21 -0.60 -0.33 -0.33 -0.29 0.34 0.0 -0.04 

*

MTAXA 0.08 -0.33 -0.24 -0.23 -0.20 -0.02 0.07 -0.12 -0.34 0.03 OLBI 0.06 0.45 0.30 0.42 0.16 0.12 0.16 -0.12 0.16 0.0 

* *

MGBI 0.17 0.22 0.0 0.05 0.29 0.29 0.25 0.02 0.22 -0.08 MGOLRAT -0.17 -0.12 -0.02 0.16 -0.19 0.03 0.08 -0.07 -0.09 0.0 MGDFRM -0.25 -0.06 0.14 -0.15 -0.19 0.09 0.14 0.0 -0.14 0.05 MGRICH 0.08 -0.33 -0.24 -0.23 -0.19 -0.02 0.07 -0.12 -0.35 0.03 TXRICH -0.08 -0.46 -0.41 -0.54 -0.41 -0.26 -0.16 0.16 -0.09 -0.23 

* * **
3.23

Table 3.3. Summary of quadrant analysis for scores of individual benthic measures and combined benthic metric f(SERM-Q: ERM-Q=concentration of a chemical in sediment sample/ERM for that chemical) as a score of chemicaquadrant score is the score which maximized the number of points in quadrants which would be considered "correcthe incidence of false negative error (low chemical concentrations and benthos adversely impacted) and false positiconcentrations and benthos not adversely impacted as established in Canfield et al. 1996).

Number of Benthic Benthos Error- Olig. Biotic Midge-Olig Midge Midge Midge Indices in Score Score SERM-Q Index Ratio Biotic Index Deformities Richne

 (%)

1 80 59 X1 80 35 X1 80 35 X1 75 19 X1 81 34 X1 75 34 2 80 28 X 3 81 18 X X 3 81 15 X X 3 81 24 X X 3 80 19 X X XTotal 1 4 5 1 4 
3.24

Table 3.4. Summary of Sediment Quality Triad data. A plus (+) for chemistry indicates a concentration of contamQ score of >39. A plus (+) for laboratory toxicity is based on a Hyalella azteca 28-d growth and survival score of >based on a combined metric of midge-oligochaete ratio, midge taxa richness and total taxa richness score of >81. 

 %

 POOL OF

CHEMISTRY TOXICITY BENTHOS NUMBER SAMPLE POSSIBLE CON

+ + + None 0 Evidence of conta

- - - All pools 88 No evidence of co

except 4, 7 degradation

and 26

+ - - None 0 Contaminants not 

- + - 26 4 Chemicals not me

 with potential to c

- - + 7 4 Benthos response 

+ + - None 0 Contaminants may 

- + + None 0 Unmeasured chem

 causing degredati

+ - + 4 4 Chemicals not bio

 due to chemistry
3.25

Appendix 3.1. Mean abundance data (number/m ) for Oligochaeta taxa. DDIG - Dero digitata; BSWRE - Branchiura so

2

moldaviensis; LHOFF - Limnodrilus hoffmeisteri; LUDEK - L. udekemianus; LCER - L. cervix; LCLAP - L. claparedei

IBIFID - Immature with bifid setae; VFULL - Varichaetadrilus fulleri; IHRPCNS - Immature with hair/pectinate (nor

Immature with hair/pectinate (enlarged setae); TOTAL - total number of individuals; NTAXA - total number of taxa

SAMPLE

TAXA SCC 1c 2c 4c 5c 6c 7c 8c 9c 10c 11c 12c 13c 14c 15c 16c 18c 19c 20c 

DDIG 0 0 0 0 0 0 0 0 0 71 0 0 0 0 0 0 0 0 0 BSWRE 0 0 67 0 0 0 167 48 56 71 163 444 1,412 125 56 0 83 278 36 PMOLD 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 12 LHOFF 67 0 0 0 0 194 0 190 167 429 140 0 471 375 0 152 0 2,222 274 LUDEK 0 83 0 0 0 28 0 0 0 0 23 0 59 0 167 212 0 0 71 LCER 0 0 133 0 0 0 0 95 0 0 140 0 0 250 0 30 0 0 167 LCLAP 67 000000000000000000LMAUM 0 0 0 143 0 0 0 143 0 0 0 0 0 250 0 212 0 0 12 IBIFID 733 0 1,067 476 0 639 333 952 778 1,286 512 111 882 2,938 5,611 1,152 1,375 9,556 369 VFULL 0 0 0 0 0 0 0 0 0 0 628 0 0 0 0 0 0 0 583 IHRPCNS 0 0 0 0 63 0 0 190 0 0 23 0 0 0 2500 0 83 0 0 IHRPCES 0 0 0 0 0 0 0 95 0 0 0 0 0 188 2444 30 0 56 0 TOTAL 867 83 1,267 619 63 861 500 1,714 1,000 1,857 1,628 556 2,824 4,12510,7781,788 1,54212,1111,52NTAXA 2121122523623545337
3.26

Appendix 3.2. Mean abundance data (number/m ) for Chironomidae taxa. ABL - Ablabesmyia; COLTP - Coelotanypus; PRO - Procladius; TP - Tanypus; CRI - Cric

2

Chironomini; CHI - Chironomus; CRYP - Cryptochironomus; ENCHI - Endochironomus; GLPTP - Glyptotendipes; HARN - Harnischia; LIPNL - Lipinella; MICHI Parachironomus; PLUTB - Paralaurterborniella; POLY - Polypedilum; TRIB - Tribelos; STEMP - Stempellinella; TOTAL - total number of individuals; NTAXA - t

SAMPLE

TAXA SCC 1c 2c 4c 5c 6c 7c 8c 9c 10c 11c 12c 13c 14c 15c 16c 18c 19c 20c 

ABL 0 0 267 0 0 28 0 48 0 0 23 444 118 0 0 0 0 1,444 12 COLTP 0 0 0 0 0 0 0 0 333 0 0 0 235 0 0 0 0 222 0 PRO 533 0 533 0 125 139 0 333 278 71 23 56 59 0 500 0 42 278 0 TP 0 0 0 0 0 0 0 0 0 143 0 0 0 0 0 0 0 0 0 CRI 0 0 0 0 0 0 0 0 0 929 0 0 0 0 111 0 0 0 0 EPCLD 0 0 0 0 0 0 0 0 0 0 0 167 235 0 0 0 0 0 0 CRNI 0 0 0 0 0 0 0 0 56 214 0 111 0 0 111 30 42 0 0 CHI 1,067 0 0 95 63 167 0 857 0 143 628 1,111 1,118 1,125 8,056 2,000 3,625 3,611 0 CRYP 0 83 0 0 0 28 0 0 56 0 0 0 235 0 56 30 83 111 83 ENCHI 0 0 0 0 0 0 0 0 0 429 0 0 0 0 0 0 0 0 0 GLPTP 0 0 0 0 0 0 0 0 0 71 0 0 0 0 0 0 0 0 0 HARN 0 0 0 0 0 0 0 0 0 0 23 0 0 0 0 0 0 0 0 LIPNL 0 0 0 0 0 28 0 0 0 0 0 0 0 0 0 0 0 0 0 MICHI 0 0 67 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 PARCHI 0 0 0 0 0 0 0 0 0 71 0 0 0 0 0 0 0 0 0 PLUTB 0 0 133 0 0 0 0 48 0 0 0 0 0 0 0 0 0 0 0 POLY 0 0 67 0 0 0 0 0 0 0 23 389 0 0 56 30 42 0 0 TRIB 67 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 STEMP 0 83 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 TOTAL 1,733 167 1,067 95 188 417 0 1,381 722 2,071 744 2,278 2,000 1,125 8,889 2,091 3,833 5,667 95 NTAXA 3 2 5 1 2 5 0 4 4 8 5 6 6 1 6 4 5 5 2 
3.27

Appendix 3.3. Mean abundance data (number/m ) for benthic invertebrate taxa (excluding oligochaetes and chironomids). HIRUDINEA: HSTAG - H

2

GPSEUD - Gammarus pseudolimnaeus; HYDRACHNIDA: UNICOLA - Unionicola; EPHEMEROPTERA: BRACH - Brachycercus; HBILIM - HexaDSPIN - Dromogomphus spinosus; HEMIPTERA: CORIX - Corixidae; TRICHOPTERA: HBDNS - Hydropsyche bidens; PFLVA - Potomyia flava; Nectopsyche; OCTS - Oecetis; TRICP - Trichoptera Pupae; DIPTERA: CRATO - Ceratopogon; PALPO - Palpomyia; PROBEZ - Probezzia; HEMRSPHAE - Sphaeriidae; MTRANS - Musculium transversum; TOTAL - total number of individuals; NTAXA - total number of taxa.

SAMPLE

TAXA SCC 1c 2c 4c 5c 6c 7c 8c 9c 10c 11c 12c 13c 14c 15c 16c 18c 19c 20c 

HSTAG 0 0 0 0 0 0 0 0 333 0 0 56 706 0 0 0 0 0 0 GPSEUD 0 0 0 0 0 28 56 0 833 0 0 56 353 0 0 0 0 0 0 UNICOLA 0 0 0 48 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 BRACH 0 0 0 0 0 56 0 0 0 0 0 0 0 0 0 0 0 0 0 HBILIM 0 0 600 95 125 28 944 0 1,889 0 140 2,667 412 0 222 0 0 3,278 36 DSPIN 0 0 0 0 0 0 0 48 0 0 0 0 0 0 0 30 0 0 12 CORIX 0 0 0 0 0 28 0 0 0 0 0 56 0 0 0 0 0 0 0 HBDNS 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 PFLVA 0 0 0 0 0 0 0 0 0 0 23 0 0 0 0 0 0 0 12 NCTO 0 0 0 0 0 0 0 0 56 0 0 0 0 0 0 0 0 0 0 OCTS 0 0 0 0 63 0 0 0 56 0 0 0 0 0 0 0 0 0 0 TRICP 0 0 0 0 0 0 56 0 0 71 0 0 0 0 0 0 0 0 0 CRATO 0 0 0 0 0 56 0 0 0 0 0 0 0 0 0 0 0 0 0 PALPO 0 0 0 0 0 0 0 0 0 0 0 0 0 63 0 0 0 0 0 PROBEZ 0 0 0 0 0 56 0 95 0 0 0 167 0 63 111 0 0 0 0 HEMRO 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 12 SPHAE 67 0 0 0 125 0 0 0 0 71 23 0 0 125 0 0 0 0 12 MTRANS 0 0 0 0 0 0 2,611 0 16,667 0 0 1,667 1,000 0 167 0 167 1,333 0 TOTAL 133 0 600 2,667 313 278 3,722 905 19,833 143 209 5,000 2,471 250 556 30 208 4,611 83 NTAXA 2 0 1 3 3 7 5 4 6 2 4 7 4 3 4 1 2 2 5