20070812 Ver 1_01 RETEC Group Report_20080502

Characterization of the Toxicity
and Bioavailability of Polycyclic
Aromatic Hydrocarbons in
Aquatic Sediments from Badin
Lake
Badin, North Carolina
Prepared by:
The RETEC Group, Inc.
1001 W. Seneca St., Suite 204
Ithaca, NY 14850-3342
RETEC Project Number: ALC02-18801-400
Prepared for:
Aluminum Corporation of America
100 Technical Drive
Alcoa Center, PA 15069
February 16, 2007
Characterization of the Toxicity
and Bioavailability of Polycyclic
Aromatic Hydrocarbons in
Aquatic Sediments from Badin
Lake
Badin, Forth Carolina
Prepared by:
The RETEC Group, Inc.
1001 W. Seneca St., Suite 204
Ithaca, NY 14850-3342
RETEC Project Number: ALC02-18801-400
Prepared for:
Aluminum Corporation of America
100 Technical Drive
Alcoa Center, PA 15069
Prared by:.,
Nie Azzolina, Hydrogeologist
r
. K-?) 1/,, ""17;
P. Kreitinger; Senor Toxicologist
February 16, 2007
Executive Summary
Alcoa, Inc. sponsored a voluntary research project to assess the bioavailability and
toxicity of polycyclic aromatic hydrocarbons (PAHs) in sediments of Badin Lake,
North Carolina adjacent to the Alcoa Badin Works (an aluminum smelter). An
investigation of Badin Lake sediment quality had previously determined that a small
embayment at the south end of the lake adjacent to the smelter had been impacted
with PAHs from historic smelter operations. The goals of this research project were
two-fold. The first goal was to determine if the PAHs in the sediments are
bioavailable and toxic to benthic aquatic organisms, and the second goal was to
evaluate whether the measurement of PAHs in sediment pore water using solid phase
microextraction (SPME) or rapidly released PAHs in sediment using supercritical
fluid extraction (SFE) could be used to predict PAH bioavailability and toxicity. The
project was designed to build upon the information and database for PAH
bioavailability and toxicity in freshwater sediments that has been initiated by Alcoa,
National Grid, and the Northeast Gas Association.
For this project, a set of sediment samples was collected from the Badin Works site
and analyzed for various biological, chemical, and physical properties. Mean
concentrations of total extractable EPA priority pollutant parent PAHs (PAH16)
ranged from 0.23 to 1,690 mg/kg in the sediment samples tested. The concentrations
of 18 parent PAH compounds plus 16 groups of alkylated PAHs were also measured
(PAH34). Sediment samples having concentrations of total extractable PAH16 greater
than 1.0 mg/kg were determined to have only modest concentrations of alkylated
PAHs indicating that the primary source of PAHs in sediments was pyrogenic in
origin, consistent with emissions from former smelting and carbon plant operations.
The ratio of total extractable parent PAHs (PAH16) to the parent plus alkylated PAH
compounds (PAH16/PAH34) was very consistent ranging from 0.46 to 0.65.
Some of the sediment samples tested were found to have high concentrations of total
organic carbon (TOC) with one sample having a carbon content of 42%. The carbon
present in sediment samples having TOC content greater than 4% or PAH
concentration greater than 10 mg/kg PAH16 consisted primarily of heat stable carbon.
These samples were measured to have a heat stable carbon/TOC ratio greater than
50%. These data suggest that the primary form of carbon in the samples has an
anthropogenic origin consistent with emissions from the smelter and associated
carbon plant and the use of anthracite coal, coal based coke, and coal tar paste at the
smelter.
The mean concentration of PAH34 in sediment pore water determined by SPME
ranged from less then detection to 76.7 µg/L. When expressed in terms of toxic units
to characterize PAH bioavail ability, these samples ranged in pore water
concentrations of PAHs from 0 to 11 TU34. The mean concentration of SFE rapidly
released PAHs (PAH34) ranged from 0.05 to 5.48 mg/kg. When expressed in terms of
toxic units, the concentration of rapidly released PAHs ranged from 0.0 to 0.7 TU34.
Measurement of bioavailable PAHs using SPME and SFE in sediment samples
collected from other aluminum smelter and manufactured gas plant (MGP) sites
E-1
Executive Summary
indicate that toxic effects to the benthic aquatic invertebrate Hyalella azteca in 28-day
laboratory tests are not expected in samples having less than 6.7 and 0.6 TU34,
respectively. These data from other sites also demonstrate that sediments samples
having more than 32 and 19 TU34 as determined by using the SPME and SFE
methods, respectively, are expected to result in reduced H. azteca survival. None of
the sediment samples collected from Badin Works exceeded the concentrations at
which H. azteca would be expected to have reduced survival.
The bioavailability and toxicity of PAHs was determined directly in sediment
samples from Badin Lake by measuring survival and growth of the aquatic amphipod
H. azteca and the aquatic midge Chironomus tentans. The survival and growth of H.
azteca were measured in 28-day chronic toxicity tests and the survival and growth of
C. tentans were measured in 10-day acute toxicity tests. Survival of H. azteca and C.
tentans in the control sediment was 95 ± 8.4 and 92.9 ± 11.1%, respectively. No
significant reduction in the survival of amphipods (H. azteca) was observed for
animals exposed to any of the sediments tested. This included exposures to sediment
BL06 which had 1,690 mg/kg total PAH16. The survival of animals exposed to
sediment BL06 was 75% ± 19%. Significantly reduced growth of H. azteca was
observed in animals exposed to one sediment sample, BL09 (0.13 ± 0.02
mg/organism), compared to the fine-grained field reference sediment, BL21 (0.24 ±
0.07). However, growth of animals exposed to sediment BL09 was not significantly
different than the growth observed in animals exposed to the laboratory control
sediment (0.19 ± 0.04), and there was no dose-response observed between the
concentration of PAHs (total extractable, SPME, or SFE) and organism growth.
No significant reduction in survival of C. tentans was observed in 10-day acute
toxicity tests compared to the survival of animals exposed to the laboratory control
sediment (92.9 ± 11%). However, a significant reduction in survival was observed
for animals exposed to sediment BL11 (62.9 ± 28.1) when compared to the fine-
grained field reference sediment BL21 (100 ± 0). No dose response was observed
between the concentration of PAHs (total extractable, SPME, or SFE) and organism
survival. There were no statistically significant differences in the growth of C. tentans
exposed to the test sediments during the 10-day acute toxicity test.
This characterization of toxicity and bioavailability of PAHs in aquatic sediments
from Badin Lake indicate that the PAHs present in sediment samples do not represent
a significant source of toxicity to benthic aquatic organisms. The two analytical
methods (SFE and SPME) currently being developed to screen sediment samples for
the concentration of bioavailable PAHs accurately predicted the absence of toxicity to
both the aquatic amphipod H. azteca and aquatic midge C. tentans. Sediment
samples which have the highest concentrations of TOC are also the same samples
which have the highest concentration of heat-stable carbon and highest concentration
of total extractable PAHs. The high carbon content present in these sediment samples
appears to greatly reduce the bioavailable concentration of PAHs and prevent
potential toxicity resulting from PAHs to aquatic life.
E-2
Table of Contents
1 Introduction ................................................................................................ .... 1-1
2 Field Activities ........................................................................................... .... 2-1
2.1 Sample Station Location ................................................................ .... 2-1
2.2 Field Measurements ....................................................................... .... 2-1
2.3 Sediment Sample Collection and Processing ................................. .... 2-2
3 Sediment Analyses ..................................................................................... .... 3-1
3.1 Sample Screening and Selection for Detailed Characterization .... .... 3-1
3.2 Analyses and Testing Methods ...................................................... .... 3-1
4 Results and Interpretation .......................................................................... .... 4-1
4.1 PAH Analyses ................................................................................ .... 4-1
4.2 TOC and Soot Carbon Analyses .................................................... .... 4-2
4.3 General Sediment Characteristics .................................................. .... 4-2
4.4 Cyanide .......................................................................................... .... 4-3
4.5 Characterization of Sediment Toxicity .......................................... .... 4-3
4.6 PAH Concentrations vs. Toxicity .................................................. .... 4-5
5 Conclusions ................................................................................................ ....5-1
6 References .................................................................................................. ....6-1
ALC02-18801-400 i
List of Appendices
Appendix A Digital Laboratory Data (compact disc)
Appendix B U. S. Army Corps ERDC Acute and Chronic Sediment Toxicity Final
Report for Badin Lake Sediment Exposed Chironomus tentans and
Hyalella azteca
ALC02-18801-400 ii
List of Tables
Table 2-1 Sediment Sample Latitude and Longitude Locations and Water Depth
Table 2-2 Field Water Quality
Table 3-1 Preliminary Screening Data
Table 3-2 Method References
Table 4-la Sediment Total PAH Concentrations
Table 4-lb Sediment SPME Porewater PAH Concentrations
Table 4-lc Sediment SFE PAH Concentrations
Table 4-2 General Physical and Chemical Characteristics
Table 4-3 Sediment Cyanide Concentrations
Table 4-4a H. azteca Toxicity Test Data - Survival & Growth
Table 4-4b C. tentans Toxicity Test Data - Survival & Growth
Table 4-5 Spiked Pitch and DNAPL Total PAH Concentrations
ALC02-18801-400 iii
List of Figures
Figure 2-1 Sediment Sample Location Map
Figure 4-1 Relative Distribution of Total Extractable PAHs in Badin Lake, Tar-,
and Pitch-spiked Control Sediments
Figure 4-2 PAH Concentrations Compared to Survival of H. azteca and C. tentans
Figure 4-3 PAH Concentrations Compared to Growth of H. azteca and C. tentans
Expressed as a Percent Change in Weight Compared to Laboratory
Control Sediment
Figure 4-4 Relationship Between Total Organic Carbon Content and Heat Stable
Carbon Content in Badin Lake Sediments
Figure 4-5 Relationship Between Total Extractable PAH16 Concentration and Total
Organic Carbon Content in Badin Lake Sediments
ALC02-18801-400 iv
1 Introduction
Alcoa, Inc. sponsored this voluntary research project to assess the
bioavailability and toxicity of polycyclic aromatic hydrocarbons (PAHs)
associated with Alcoa's Badin Works (an aluminum smelting facility) in
sediments of Badin Lake, North Carolina. Previous investigations had
determined a small embayment at the south end of the lake contained
sediments impacted with PAHs from historical operations at the smelter. The
goals of this project were two-fold. First, to determine if the PAHs in the
sediments may be bioavailable and toxic to benthic organisms within the
sediment, and second, to evaluate whether the measurement of PAHs in
sediment pore water using solid phase micro extraction (SPME) or rapidly
released PAHs in sediment using supercritical fluid extraction (SFE) could be
used to predict PAH bioavailability and toxicity. The project was designed to
build upon the information and database for PAH bioavailability and toxicity
in freshwater sediments that has been initiated by Alcoa, National Grid, and
the Northeast Gas Association.
This assessment of PAH bioavailability and toxicity was conducted following
the Project Work Plan (RETEC, 2005) with support by the following principle
contractors:
• Dr. Steven B. Hawthorne - Sediment PAH Availability Chemistry
Energy and Environment Research Center, University of North Dakota
Grand Forks, ND
• Dr. Todd Bridges/ Mr. J. Daniel Farr - Aquatic Toxicity Testing
US Army Engineer Research and Development Center
Vicksburg, MS
• Dr. Eleanor Hopke - Sediment Cyanide Analysis
Clarkson University
Potsdam, NY
• Mr. Dave Dunlap - General Sediment Chemistry
Severn Trent Laboratories (STL)
Pittsburgh, PA
A description of the field activities performed at the site, the results of the
analytical work performed, and interpretation of the data are presented in this
report.
ALC02-18801-400 1-1
2 Field Activities
Twenty-two sediment samples were collected on April 27-28, 2005, from
locations having PAH concentrations ranging from 10 to 2,100 mg/kg total
PAHs as identified during previous investigation activities conducted by
Woodward-Clyde Consultants (WCC) [Woodward-Clyde 1997]. Unless
otherwise noted below, the field work was performed using the methods and
procedures provided in the project work plans Sediment Sample Collection
and Analysis Work Plan for Evaluating PAH Bioavailability (RETEC, 2005)
and were consistent with methods specified by the United States
Environmental Protection Agency (U. S. EPA, 2001) and the American
Society for Testing Materials (ASTM, 2000a; ASTM 2000b).
2.1 Sample Station Location
Figure 2-1 shows the location of each sampling station. Table 2-1 provides
the latitude and longitude coordinates of each sampling station and water
depth at the time of sample collection. Water depth was measured with a
weighted tape measure with an accuracy of 0.1 feet.
Three samples, BL20, BL21, and BL22, were collected as potential far field
reference sediment samples for toxicity testing. These samples were located
approximately 3 miles northeast of the Badin Smelter site (Figure 2-1).
Samples BL20 and BL21 were selected as coarse- and fine-grained field
reference samples, respectively.
Field personnel recorded observations of the physical characteristics of the
sediment encountered at each sampling station, and also important
observations regarding the physical characteristics of the study area.
Information recorded included the presence of fill materials at the shoreline;
apparent depositional and erosional environment at the station; presence or
absence of aquatic vegetation; sediment color, texture and particle size; and
odor and presence of sheens or non-aqueous phase liquids (NAPL) [Table 2-
1].
2.2 Field Measurements
Water quality was measured in the field at each sample station location (Table
2-2). Two sets of water quality measurements were taken at each station.
One measurement was taken near the water surface, approximately 0.5 meter
below the water depth, and a second measurement was taken near the bottom,
approximately 0.5 meter above the top of the sediment surface. The water
quality parameters measured in the field were temperature, salinity, dissolved
oxygen (DO), conductivity, and pH.
ALC02-18801-400 2-1
Characterization of the Toxicity and Bioavailability of Polycyclic Aromatic Hydrocarbons in Aquatic
Sediments from Badin Lake - Badin, North Carolina
2.3 Sediment Sample Collection and
Processing
Approximately 4 gallons of surface sediment (top 6 inches) were collected
from each sample station using a Ponar grab sampler or a trowel/shovel where
the water depth above the sediment was less then 2 feet deep and near the
shoreline.
At each station, sediments were initially screened with a #3/8-mesh (9.5-cm
openings) screen to remove large objects such as cobbles, sticks, or shells.
The screened sediments were then placed into a clean, 5-gallon, plastic
bucket. The buckets were labeled with the sample station designation and
transported to an on-shore sample preparation area where the samples were
homogenized by screening to < 4 mm and mixed to ensure the consistency of
subsamples sent to each laboratory.
ALC02-18801-400 2-2
3 Sediment Analyses
3.1 Sample Screening and Selection for
Detailed Characterization
Twenty-two sediment samples were collected from the site, including a
coarse- and a fine-grained field reference sample. Sediment samples were
initially screened for their PAH concentration, total organic carbon (TOC),
and heat stable organic carbon (SOC) content. This preliminary data on PAH
concentration and the TOC and SOC content was used to guide the selection
of a subset of 12 samples for detailed characterization of PAH bioavailability.
The preliminary analysis of the total concentration of sixteen EPA priority
pollutant PAHs (PAH16) showed that the samples collected ranged from
approximately 0.1 to 901 mg/kg (wet weight), and the concentration of TOC
ranged from 0.3% to 42.4%. For samples having total concentrations of
PAH16 more than 1.0 mg/kg wet wt, the ratio of parent to total PAH
compounds (PAH16/PAH34) was very consistent ranging from 0.63 to 0.84.
The ratio of heat stable carbon/TOC varied from 0.19 to 1.00, respectively
(Table 3-1).
In addition to a coarse- and fine-grained field reference sample for toxicity
testing, BL 18 was selected as a near-field reference sediment based on its low
concentration total PAH16 (1.8 mg/kg wet wt, Table 3-1) measured by the
preliminary analysis of PAHs.
3.2 Analyses and Testing Methods
Table 3-2 provides method references for the testing and the laboratories
conducting the analysis. A detailed description of the test methods and
rationale is provided in the project work plan.
ALC02-18801-400 3-1
4 Results and Interpretation
Chain of custody (COC) documents, laboratory results, and field data sheets
are included in Appendix A as electronic portable data files (pdf) on compact
disc (CD).
4.1 PAH Analyses
Based on the preliminary analysis for PAHs, 12 samples were selected for
characterization of PAH bioavailability and toxicity. These samples were
selected to have a wide range in total extractable PAHs and were measured to
have 0.23 to 1,690 mg/kg total PAH16 (dry wt., Table 4-1a). Samples BL20
and BL21, selected as coarse- and fine-grained field reference samples for
toxicity testing, were determined to have 0.23 and 0.76 mg/kg PAH16,
respectively. Sample BL18, the sample selected as a near field reference
sample, was determined to have 5.2 mg/kg PAH16.
In addition to measuring the concentration of 16 EPA priority pollutant PAHs,
samples were analyzed for the concentration of 34 NOAA PAHs (U.S. EPA,
2000) which consist of 18 parent PAHs and 16 groups of alkylated PAHs
(PAH34). For sediment samples having concentrations of PAH16 greater than
1.0 mg/kg, the ratio of 16 EPA PAHs to 34 NOAA PAHs (PAH16/PAH34) was
low and very consistent ranging from 0.46 to 0.65 (Table 4-1a). These data
indicate that the primary source of PAHs in sediments was pyrogenic in origin
(i.e. resulting from combustion processes) and consistent with emissions from
the smelter and associated carbon plant.
Field observations of NAPL sheens and the presence m/z ions, characteristic
of alkanes in the mass spectral analysis of PAHs, indicated that a petrogenic
source of hydrocarbons (i.e. resulting from petroleum) may also be present in
several sediment samples collected from Badin Lake. However, the NAPL
present in some samples did not appear to contribute significantly to the
concentration of alkylated PAHs or the toxicity of the sediment (see section
4.6).
The concentration of dissolved PAHs in sediment pore water, measured using
SPME, was used as a predictor of PAH bioavailability and toxicity. The mean
pore water concentration of PAH34 ranged from below detection to 76.7 µg/L
(Table 4-1b). In addition to expressing the dissolved concentration of PAHs in
pore in terms of ug/L, the pore water PAH concentration is also expressed in
terms of toxic units (TU34) which is the sum of each measured pore water
PAH concentration divided by its EPA-derived final chronic value (FCV). A
toxic unit (TU) is a hazard quotient that normalizes the measured
concentration of PAHs in sediment or pore water to the concentration of PAH
expected to result in toxicity to sensitive macroinvertebrates. For a mixture of
PAHs, the TUs for individual parent and groups of alkylated PAHs are
summed. The total TUs (sum of the TUs for the mixture of 34 PAHs)
ALC02-18801-400 4-1
Characterization of the Toxicity and Bioavailability of Polycyclic Aromatic Hydrocarbons in Aquatic
Sediments from Badin Lake - Badin, North Carolina
measured in pore water by SPME ranged from <0.1 to 11 TU34 (Table 4-1b).
Data developed from multiple aluminum industry and manufactured gas plant
(MGP) sites indicates that pore water samples having less than approximately
6.7 TU34 are not anticipated to result in significantly reduced survival (> 85%
survival) of H azteca and samples having more then 32 TU34 are expected to
have near 100% mortality (<15% survival). Additional discussion on the
concentration of PAHs measured in sediment pore water and the results of
toxicity tests are presented in Section 4.6.
The mean concentration of SFE rapidly released PAHs (PAH34) ranged from
0.05 to 5.48 mg/kg (Table 4-1c). When expressed in terms of toxic units,
these samples had SFE rapidly released PAH concentrations ranging from
<0.1 to 0.7 TU34 (Table 4-1b). Sediment samples having less than
approximately 0.6 TU34, as determined by SFE, are not anticipated to result in
significantly reduced survival of H azteca and sediment samples having more
then 19 TU34 are expected to have near 100% mortality (<15% survival).
Additional discussion on the concentration of rapidly released PAHs
measured by SFE and the results of toxicity tests are presented in Section 4.6.
4.2 TOC and Soot Carbon Analyses
TOC and SOC were measured using a CHN elemental analyzer. Heat stable
organic carbon is defined as the TOC remaining following pretreatment at 375
°C in a muffle furnace under oxidizing conditions (Table 4-2). Many of the
samples had high concentrations of TOC with one sample having a carbon
content of 42%. The carbon present in sediment samples having a high TOC
content (> 4.0%) or high PAH concentration (> 10 mg/kg PAH16) consisted
primarily of heat stable carbon (SOC/TOC ratio > 0.50).
Samples BL20 and BL21, selected as coarse- and fine-grained field reference
samples, were determined to have 0.3 and 2.3% TOC, respectively. Sample
BL 18, the near field reference sample, was determined to have 3.7% TOC.
4.3 General Sediment Characteristics
Additional analyses of sediment grain size, pH, ammonia, and percent solids
were also performed (Table 4-2). The complete digital laboratory report from
STL is provided in Appendix A.
Sediment grain size varied from predominantly sand with 10-20% silt to silts
and clays with less than 10% sand. Samples BL20 and BL21, selected as a
coarse- and fined-grained field reference samples were determined to have 88
and 1.1% sand, respectively. Sample BL18, the sample selected as a near
field reference sample, was determined to have 51% sand.
ALC02-18801-400 4-2
Characterization of the Toxicity and Bioavailability of Polycyclic Aromatic Hydrocarbons in Aquatic
Sediments from Badin Lake - Badin, North Carolina
The pH of all sediment samples was circum-neutral, between 6.5 and 6.9.
Concentrations of ammonia varied from 13 to 109 mg/kg (Table 4-2).
4.4 Cyanide
Total, free, and iron-complexed cyanide was measured in the sediment
samples used for toxicity testing. Cyanide is a potential by-product from
aluminum smelting operations, and free cyanide is a potential toxicant and
confounding factor in the interpretation of toxicity testing results. The
complete digital laboratory report from Clarkson University is provided in
Appendix A.
The concentration of free cyanide was less than 0.25 mg/kg in all of the
sediment samples tested (Table 4-3). The highest concentration of free
cyanide (0.50 mg/kg) was measured in the field reference sample, BL21,
collected from an unimpacted area of Badin Lake. The aquatic life criterion
for free cyanide in water is 5.2 µg/L (U. S. EPA, 1996). The concentration of
total cyanide and iron cyanide ranged from <0.05 to 6.6 and <0.006 to 3.8
mg/kg, respectively. Based on these data, no toxicity to aquatic invertebrates
was anticipated in laboratory toxicity test.
4.5 Characterization of Sediment Toxicity
The bioavailability and toxicity of PAHs was determined directly by
measuring the toxicity of sediment to aquatic amphipods and midges. The
survival and growth of the amphipod H. azteca was measured in 28-day
chronic toxicity tests (Table 4-4a) and the survival and growth of the midge C.
tentans was measured in 10-day acute toxicity tests (Table 4-4b). The
survival and growth of animals exposed to the test sediments was compared to
a corresponding coarse-grained (BL20) or fine-grained (BL21) field reference
sediment. In addition, a near-field reference sediment (BL 18) was used to
expose animals to a sediment sample collected from the site which had a low
concentration of PAH16 (5.24 mg/kg). All toxicity testing was performed by
the U. S Army Engineer Research and Development Center (ERDC). The
complete digital laboratory report from ERDC is provided in Appendix A.
Survival of H. azteca and C. tentans in the uncontaminated laboratory control
sediment (negative control sediment) was 95 ± 8.4 and 92.9 ± 11.1%,
respectively. No significant reduction in the survival of amphipods (K
Azteca) was observed for animals exposed to any of the test sediments. This
included exposure to sediment BL06 which had 1,690 mg/kg total PAH16.
The survival of animals exposed to BL06 was 75 ± 19%. Significantly
reduced growth of H. azteca was observed in animals exposed to one
sediment sample, BL09 (0.13 ± 0.02 mg/organism), compared to the fine-
grained field reference sediment, BL21 (0.24 ± 0.07). However, growth of
animals exposed to sediment BL09 was not significantly different than the
growth observed in animals exposed to the laboratory control sediment (0.19
ALC02-18801-400 4-3
Characterization of the Toxicity and Bioavailability of Polycyclic Aromatic Hydrocarbons in Aquatic
Sediments from Badin Lake - Badin, North Carolina
± 0.04, Table 4-4a). The concentration of total PAH16 in sediment sample
BL09 was 109 ± 102 mg/kg and the concentration of SFE rapidly available
PAH and pore water PAH concentration in BL09 was less than 0.1 TU34
(Tables 4-lb and 4-1c).
No significant reduction in survival of C. tentans was observed in 10-day
acute toxicity tests compared to the survival of animals exposed to the
laboratory control sediment (92.9 ± 11%). However, a significant reduction in
survival was observed for animals exposed to sediment BL 11 (62.9 ± 28.1%)
when compared to the fine-grained field reference sediment BL21 (100 ± 0%).
The concentration of total PAH16 in sediment sample BL11 was 80.0 ± 3.2
mg/kg and the concentration of SFE rapidly available PAH and pore water
PAH concentration was 0.1 ±0.1 and 2.2 ± 0.1 TU34, respectively (Tables 4-lb
and 4-1c). There were no statistically significant differences in the growth of
C. tentans exposed to the test sediments during the 10-day acute toxicity test
(Table 4-4b).
To evaluate the toxicity of bioavailable PAHs similar to the PAH mixture
observed in field collected sediments, a positive control treatment was
prepared by adding either a coal tar pitch (Pitch) or fresh coal tar (Tar)
solution prepared in acetone to the laboratory control sediment. The final
concentration of total extractable PAH16 was 55.6 and 44.3 mg/kg for the
pitch- and tar-spiked positive control treatments, respectively (Table 4-5).
Upon examination of the distribution of low molecular weight to high
molecular weight PAHs, it was apparent that the pitch-spiked positive control
sediment was more similar to the contaminated Badin Lake sediment samples
(i.e. samples with > 100 mg/kg PAH16) than the distribution of PAHs in the
fresh tar-spiked control sediment (Figure 4-1). The tar-spiked control
sediment had higher relative concentrations of low molecular weight PAHs
(i.e. naphthalenes, acenaphthylene, acenaphthene, and fluorenes) when
compared to the field collected sediments.
The survival of amphipods in the pitch-spiked sample was greatly reduced
(6.7 ± 16.3%), while the survival of amphipods was unaffected in the tar-
spiked sample (93.3 ± 12.1) (Table 4-4b). In contrast, the survival of
chironomids was significantly reduced in both pitch-spiked and tar-spiked
control sediments compared to the laboratory control and field reference
samples (Table 4-4b).
The lack of toxicity to amphipods in the tar-spiked control sediment was
unexpected; however, other researchers have reported an unexpected lack of
toxicity to H. azteca in sediment samples that were spiked with hydrophobic
organic compounds (Call et al. 2001). This lack of toxicity has been
attributed to avoidance behavior of H. azteca which is an epibenthic species
that is able to swim in the water column above the sediments thus potentially
reducing its exposure during the toxicity test. In addition, the lack of toxicity
ALC02-18801-400 4-4
Characterization of the Toxicity and Bioavailability of Polycyclic Aromatic Hydrocarbons in Aquatic
Sediments from Badin Lake - Badin, North Carolina
has also been attributed to a loss of the test chemical from dissolution and the
daily renewal of the overlying water, as well as volatilization. Low molecular
weight PAHs (naphthalenes, acenaphthylene, acenaphthene, and fluorenes)
have higher solubility and vapor pressure compared to higher molecular
weight PAHs and would be more subject to these losses than higher molecular
weight PAHs.
Observations of H. azteca swimming above the sediment surface during the
tests were noted for the tar-spiked sediment but not the pitch-spiked treatment.
Thus it appears that a lack of toxicity in the tar-spiked control sediment may
have occurred as a result of the reduced exposure to H. azteca and potential
loss of the low molecular weight PAHs from volatilization and renewal of the
overlying water during the test. The Badin Lake sediment samples were
similar in PAH composition to the pitch-spiked control sediment and thus this
confounding effect on the interpretation of sediment toxicity test results was
not a concern for the test samples. It is also interesting to note that the
animals surviving exposure to the tar-spiked sediment were observed to have
the highest dry weight (not statistically significant) compared to the other
treatments and laboratory control sediments. The increased growth may have
resulted from an increase in microbial populations in the tar-spiked sediment
samples and a subsequent increase in the nutritional quality of the tar spiked-
sediment.
Based on these data, it is recommended that results from H. azteca toxicity
tests be reviewed with caution when using sediments that have a high
concentration of naphthalene and other low molecular weight PAHs. The
sediment samples collected from the Badin Lake did not have high
concentrations of naphthalene and other low molecular weight PAHs. In
addition, avoidance behavior was not observed in the toxicity tests using
sediments collected from Badin Lake.
4.6 PAH Concentrations vs. Toxicity
Despite a wide range in concentration of total extractable PAHs and a
maximum observed PAH16 concentration of 1,690 mg/kg there was no
relationship between the concentration of PAHs and the survival and growth
of amphipods or chironomids exposed to Badin Lake sediments (Figures 4-2
and 4-3). No dose response was observed between the concentration of PAHs
(total extractable, SPW or SFE) and organism survival or growth. In
contrast, sediment spiked with coal tar pitch elicited a toxic response to both
amphipods and chironomids at a concentration of only 55.6 mg/kg PAH16.
Amphipods survived exposure to sediment samples having a dissolved
concentration of pore water PAHs up to 11 TU34. Previous assessments on the
bioavailability of PAHs in sediments collected from other sites have
demonstrated that reduced survival of H. azteca is not observed in sediment
samples having dissolved PAH concentrations in pore water less than
ALC02-18801-400 4-5
Characterization of the Toxicity and Bioavailability of Polycyclic Aromatic Hydrocarbons in Aquatic
Sediments from Badin Lake - Badin, North Carolina
approximately 6.7 TU34 (Figure 4-2b). The concentration of SFE rapidly
released PAHs in Badin Lake sediment samples did not exceed 0.7 TU34.
Toxicity has not been observed in sediment samples from other sites having
less than approximately 0.6 SFE TU34 (Figure 4-2c). These results showing a
lack of toxicity in Badin Lake sediment samples are consistent with the
previous observations from other MGP and aluminum industry sites.
It appears that anthropogenic sources of carbon are responsible for the low
bioavailability and toxicity of PAHs observed in Badin Lake sediments.
Sediment samples collected from the site having high concentrations of PAHs
were also observed to have the high concentrations of TOC (Figure 4-4). In
addition, sediment samples having high concentrations of TOC were also
associated with high concentrations of SOC. These data suggest that PAHs in
sediments at the site tend to be associated with anthropogenic carbon which
has higher sorption capacities compared to natural organic carbon and thus
limiting PAH bioavailability and toxicity.
It is interesting to note that alkylated PAHs contribute the majority of toxic
units in the calculation of SPME TU34 and SFE TU34. In samples having
more than 100 mg/kg total extractable PAH34 (BL05, BL06, BL08, BL09
BL 10, BL 11, BL 12), the alkylated PAHs contribute 53 to 99% of the toxic
units in sediment pore water (SPME TU34) and 35 to 95% of the rapidly
released toxic units (SFE TU34). This suggests that alkylated PAHs are likely
to be the primary contributor to the potential toxicity of sediment samples at
primary aluminum smelter sites, despite the dominance of a pyrogenic PAH
source signature (PAH16/PAH34 ratio 0.46-0.65; Table 4-1a).
ALC02-18801-400 4-6
5 Conclusions
This characterization of toxicity and bioavailability of PAHs in aquatic
sediments from Badin Lake indicates that the PAHs present in sediment
samples do not represent a significant source of toxicity to benthic aquatic
organisms. The two analytical methods (SFE and SPME) currently being
developed to screen sediment samples for the concentration of bioavailable
PAHs accurately predicted the absence of toxicity to both the aquatic
amphipod H. azteca and aquatic midge C. tentans. Sediment samples which
have the highest concentrations of TOC are also the same samples which have
the highest concentration of heat-stable carbon and highest concentration of
total extractable PAHs. The high carbon content present in these sediment
samples appears to greatly reduce the bioavailable fraction of PAHs and
prevent potential toxicity resulting from the PAHs to aquatic life.
ALC02-18801-400 5-1
6 References
ASTM, (ed.) 2000a. E. 1391-94 Standard guide for collection, storage,
containerization, and manipulation of sediments for toxicological testing, Vol.
11.04. American Society for Testing Materials, Conshohocken, PA.
ASTM, (ed.) 2000b. D 3976 - 92 (Reapproved 2001) Standard practice for
preparation of sediment samples for chemical analysis, Vol. 11.04. American
Society for Testing Materials, Conshohocken, PA.
RETEC, 2005. Sediment Sample Collection and Analysis Work Plan for Evaluating
PAH B i oavail ability, Badin Lake, North Carolina. April 25, 2005.
U. S. EPA. 1996. Updates: Water Quality Criteria Documents for the Protection of
Aquatic in Ambient Water. EPA-820-B-96-001, September 1996
U. S. EPA. 2000. Technical basis for the derivation of equilibrium partitioning
sediment guidelines (ESGs) for the protection of benthic organisms: Nonionic
organics. EPA-822-R-00-001. U.S. Environmental Protection Agency, Office
of Science and Technology, Washington D.C.
U. S. EPA. 2001. Methods for collection, storage and manipulation of sediments for
chemical and toxicological analyses: Technical manual EPA-823-B-01-002.
United States Environmental Protection Agency, Office of Water,
Washington, D.C.
Woodward-Clyde, 1997. Preliminary Sediment and Water Sampling Results from
Alcoa's Badin, NC Works.
Call, Daniel J., Cox, Dean A., Geiger, Dianne L., Genisot, Kristen I., Markee,
Thomas P., Brooke, Larry T., Polkinghorne, Christine N., VandeVenter, Fred
A., Gorsuch, Joseph W., Robillard, Kenneth A., Parkerton, Thomas F., Reiley,
Mary C., Ankley, Gerald T., Mount, David R. 2001. An Assessment of the
Toxicity of Phthalate Esters to Freshwater Benthos. 2. Sediment Exposures.
Environmental Toxicology and Chemistry. 20 Issue: 8 Pages: 1805-1815.
ALC02-18801-400 6-1
Tables
TABLE 2-1
Sediment Contaminant Bioavailability Assessment
Sediment Sample Locations
Alcoa Badin Works Site
Selected
Sample for Detailed GPS GIPS Water
(') (1) Depth Field Description
ID Chemical Latitude Longitude
(ft)
Analysis
BL01 35 24 5223 80 6 8765 19
6 BROWN SILT, some fine-medium gravel and shells, little fine
. sand and sticks. Trace sheen. No odor.
BL02 35 24 7094 80 6 8918 0.8-1.9 GRAY fine-coarse GRAVEL and organic material, some silt
and sand. No sheen. Strong septic-like odor.
BL03 35 24 5189 80 6 8579 24.6 BROWN SILT, trace shells and twigs. No sheen or odor.
BL05 35 24 7239 80 6 8410 28.6 GRAYISH-BROWN SILT, trace shells. No sheen or odor.
BL06 35 24 7194 80 6 8594 20
9 GRAY and BROWN SILT and fine sand, trace shells. NAPL
. blebs and sheen. Faint odor.
BL08 35 24 5033 80 6 8750 21
2 BROWN SILT, some shells, little fine sand, trace twigs. No
. sheen or odor.
BL09 35 24 8382 80 6 9520 8.0 BROWN SILT, some gravel, trace shells. No sheen or odor.
BL10 35 24 7234 80 6 8545 8
24 GRAYISH-BROWN SILT, trace sand and shells.No sheen or
. odor.
BL11 35 24 7401 80 6 8742 23
3 BROWN SILT, some shells, trace fine sand. No sheen or
. odor.
BL12 35 24 7398 80 6 8339 29
3 GRAYISH-BROWN SILT, trace shells. NAPL blebs and
. sheen. No odor.
BL14 35 24 8323 80 6 9254 14
8 BROWN and GRAY SILT, trace gravel and shells. No sheen
. or odor.
BROWN SILT, some shells, wood and sticks. No sheen or
BL15 35 24 8217 80 6 8940 20.1
odor.
BROWN SAND and Silt, some gravel and shells. No sheen
BL16 35 24 8392 80 6 8938 17.8
or odor.
BROWN and GRAY fine-medium GRAVEL, some shells,
BL17 35 24 9056 80 6 8954 3.2
little medium-coarse sand. No sheen or odor.
BL18 35 24 9044 80 6 9027 5
1 BROWN GRAVEL, some shells, little silt and sticks. No
. sheen or odor.
BROWN coarse-fine GRAVEL
little sand
silt and shells. No
BL19 35 24 8884 80 6 8801 12.7 ,
,
sheen or odor.
REDDISH-BROWN fine-medium GRAVEL, some fine-coarse
BL20 35 26 0087 80 5 0461 0.0 sand, little wood and shells, trace black organic material. No
sheen or odor.
BL21 35 26 2671 80 5 0169 36.8 BROWN SILT. No sheen. Organic odor.
BROWN SAND and SILT
some shells and sticks
little
BL22 35 26 76441 80 5 2386 2.8 ,
,
gravel. No sheen. Organic odor.
(1)GPS coordinates are in WGS 1984 Degrees, Minutes, Decimal-Seconds.
Table 2-1
TABLE 2-2
Sediment Contaminant Bioavailability Assessment
Water Quality Measurements
Alcoa Badin Works Site
Sample Station
Designation Water Column
Position Temperature
Degrees Celcius
Salinity (ppt) Dissolved
Oxygen (mg/L) Conductivity
(umhos)
pH
near surface 17.2 0.0 8.6 0.6 6.8
131-01 near bottom 16.6 0.0 8.2 0.6 6.8
131-02 near surface 17.8 0.0 7.9 0.1 6.7
03
131 near surface 17.2 0.0 8.9 0.1 6.9
- near bottom 16.3 0.0 8.1 0.1 6.8
05
131 near surface 16.7 0.0 7.8 0.1 6.5
- near bottom 15.7 0.0 6.7 0.1 6.5
06
131 near surface 17.1 0.0 8.6 0.1 6.9
- near bottom 16.1 0.0 8.1 0.1 6.8
08
131 near surface 17.6 0.0 9.0 0.1 6.8
- near bottom 16.7 0.0 8.4 0.1 6.8
09
131 near surface 16.2 0.0 8.2 0.1 6.3
- near bottom 15.8 0.0 7.9 0.1 6.3
BL10 near surface 16.7 0.0 8.6 0.1 6.9
near bottom 16.0 0.0 7.7 0.1 6.7
11
131 near surface 17.1 0.0 8.5 0.1 6.9
- near bottom 16.0 0.0 7.9 0.1 6.7
BL12 near surface 16.6 0.0 8.0 0.1 6.8
near bottom 16.0 0.0 7.8 0.1 6.7
BL14 near surface 16.0 0.0 7.8 0.1 6.1
near bottom 15.6 0.0 7.5 0.1 6.2
BL15 near surface 16.2 0.0 8.0 0.1 6.3
near bottom 15.9 0.0 7.7 0.1 6.4
BL16 near surface 16.2 0.0 8.1 0.1 6.5
near bottom 16.0 0.0 7.7 0.1 6.5
BL17 near surface 16.2 0.0 8.5 0.1 6.6
BL18 near surface 15.8 0.0 8.0 0.1 6.3
near bottom 15.8 0.0 7.9 0.1 6.3
BL19 near surface 16.2 0.0 8.4 0.1 6.6
near bottom 16.0 0.0 8.2 0.1 6.6
BL20 near surface NA NA NA NA NA
BL21 near surface 16.4 0.0 8.0 0.1 6.2
near bottom 15.8 0.0 7.3 0.1 6.2
BL22 near surface 16.3 0.0 8.0 0.1 6.4
NA - Insufficient amount of water present to obtain measurements.
Table 2-2
TABLE 3-1
Sediment Contaminant Bioavailability Assessment
Sediment Screening Data
Alcoa Badin Works Site

Sample
ID Selected
For
Detailed
Chemical
Analysis
???
Total PAH1s
(Z)
Total PAH34
Ratio
PAH16/PAH34
Total
Organic
Carbon
Soot
Carbon
Ratio
Soot/Total
Carbon
mg/kg Wet Wt. mg/kg Wet Wt. Dry Wt. % Dry Wt. % Soot/TOC
BL01 33.5 49.6 0.68 6.0 5.2 0.87
BL02 117 160 0.73 15.2 9.9 0.65
BL03 40.4 58.5 0.69 9.6 8.1 0.84
BL05 83.1 143 0.58 9.0 7.3 0.81
BL06 901 1390 0.65 42.4 39.7 0.94
BL08 352 475 0.74 32.2 32.1 1.00
BL09 27.9 37.8 0.74 10.9 12.0 1.10
BL10 152 223 0.68 12.2 10.3 0.84
BL11 54.3 85.5 0.63 4.8 3.3 0.69
BL12 44.8 66.6 0.67 4.4 2.9 0.65
BL14 16.3 21.7 0.75 5.0 2.6 0.52
BL15 9.3 12.5 0.74 3.0 1.1 0.35
BL16 4.0 6.2 0.64 1.4 0.3 0.19
BL17 1.7 2.0 0.84 0.7 0.2 0.31
BL18 1.2 1.5 0.81 3.7 0.7 0.18
BL19 3.5 4.3 0.81 3.5 0.7 0.19
BL20 0.1 0.1 1.00 0.3 0.2 0.50
BL21 0.3 0.4 0.72 2.3 0.2 0.09
BL22 0.1 0.2 0.53 2.2 0.1 0.03
(1) Sum of 16 EPA priority pollutant PAHs noted below with an asterisk. (2) Sum of 34 PAHs
includes 18 parent and 16 groups of alkylated PAHs: naphthalene*, C1 naphthalenes, C2
naphthalenes, C3 naphthalenes, C4 naphthalenes, acenaphthylene*, acenaphthene*, fluorene*, C1
fluorenes,C2 fluorenes, C3 fluorenes, phenanthrene*, anthracene*, C1 phenanthrenes/anthracenes,
C2 phenanthrenes/anthracenes, C3 phenanthrenes/anthracenes, C4 phenanthrenes/anthracenes,
fluoranthene*, pyrene*, C1 fluoranthenes/pyrenes, benz[a ] anthracene*, chrysene*, C1 chrysenes,
C2 chrysenes, C3 chrysenes, C4 chrysenes, benzo[b+k]fluoranthene*, benzo[e ]pyrene,
benzo[a]pyrene*, perylene, indeno[1,2,3-cd]pyrene*, dibenz[ah]anthracene*,
benzo[ghi ]perylene*.
Table 3-1
TABLE 3-2
Sediment Contaminant Bioavailability Assessment
Analytical Testing Program
Alcoa Badin Works Site
Sample Matrix Parameter Purpose Quantity Laboratory'
Type
PAH Screen - PAHs parent & alkylated (34 compounds) Data to be used to select
Initial Characterization Sediment Total Organic Carbon samples for detailed 22 EERC
Soot Carbon characterization

Amphipod toxicity (Hyalella azteca) 28-day chronic test 13: 10 field samples,
Bioavailability S
di
t Direct Measurement of PAH 2
f
l
1 ERDC
Characterization e
men bioavailability re
erence samp
es,
Midge toxicity (Chironomus tentans ) 10-day chronic test positive control

Total and rapidly available PAHs [parent & alkylated 34 compounds] Chemical test for estimating

bioavailability 12 EERC
Total Solids
Grain size General sediment
Sediment characteristics potential 12 STL
Chemical/Physical pH confounding factors affecting
Characterization interpretation of toxicity data
Ammonia
Cyanide - total, free, and iron (Fe)-complexed Potential confounding factors 12 CLKU
Pore Dissolved Organic Carbon (DOC) Chemical test for estimating
Water 12 EERC
bioavailability
Free' dissolved PAHs (parent compounds and estimate of al lated

Temperature
pH
Fi
ld M Surface Dissolved oxygen General environmental quality 22 YSI 6820 Field
e
easurement Water Conductivity parameters Meter
Salinity
Turbidity

Note 1. EERC - Energy & Environment Research Center
ERDC - Army Engineering Research and Development Center
STL - Severn Trent Laboratories, Inc., Burlington, VT
CLKU - Clarkson University
Table 3-2
TABLE 4-la
Sediment Contaminant Bioavailability Alliance
Total PAH Concentrations
Alcoa Badin Works Site
BL01 BL05 BL06 BL08 BL09 BL10
Sediment PAH's m /k Mean Std Dev. N Mean Std Dev. N Mean Std Dev. N Mean Std Dev. N Mean Std Dev. N Mean Std Dev. N
naphthalene 0.14 0.02 4 0.26 0.04 4 2.64 0.38 4 0.58 0.10 4 0.72 0.49 4 0.40 0.03 4
2-methyl nahthalene 0.12 0.02 4 0.19 0.01 4 1.91 0.29 4 0.37 0.11 4 0.45 0.27 4 0.35 0.02 4
1-meth Inaphthalene 0.07 0.01 4 0.15 0.01 4 1.30 0.17 4 0.23 0.07 4 0.26 0.22 4 0.21 0.01 4
C2 naphthalenes 0.56 0.05 4 3.38 0.69 4 10.0 1.69 4 1.32 0.17 4 1.08 0.76 4 1.99 0.09 4
C3 naphthalenes 0.29 0.04 4 4.42 1.25 4 8.84 1.83 4 0.79 0.07 4 0.48 0.34 4 2.00 0.11 4
C4 naphthalenes 0.22 0.02 4 2.20 0.57 4 3.89 0.75 4 0.44 0.02 4 0.19 0.07 4 1.00 0.10 4
acena hth lene 0.08 0.02 4 0.12 0.01 4 0.50 0.04 4 0.29 0.01 4 0.11 0.07 4 0.15 0.01 4
acena hthene 0.20 0.05 4 0.84 0.08 4 10.2 0.75 4 2.21 0.20 4 0.80 0.76 4 1.59 0.03 4
fluorene 0.17 0.03 4 0.59 0.06 4 8.07 0.89 4 1.71 0.15 4 0.70 0.68 4 1.26 0.02 4
Cl fluorenes 0.41 0.09 4 1.35 0.22 4 8.88 0.91 4 2.08 0.24 4 0.82 0.77 4 1.47 0.11 4
C2 fluorenes 0.31 0.05 4 2.60 0.70 4 7.12 0.74 4 1.34 0.12 4 0.40 0.28 4 1.46 0.09 4
C3 fluorenes ND ND ND ND ND ND ND ND ND ND ND ND ND ND ND ND ND ND
henanthrene 1.90 0.30 4 7.41 0.55 4 84.9 9.21 4 29.1 3.10 4 8.58 7.48 4 13.3 0.14 4
anthracene 0.98 0.11 4 2.69 0.25 4 34.0 3.80 4 11.2 1.39 4 2.81 2.36 4 5.89 0.22 4
Cl henanthreneslanthracenes 2.33 0.38 4 7.55 1.12 4 62.7 5.81 4 19.3 1.88 4 7.25 5.68 4 11.6 0.32 4
C2 henanthreneslanthracenes 1.46 0.19 4 22.0 5.62 4 56.1 5.27 4 19.6 1.89 4 5.86 3.95 4 33.8 2.24 4
C3 henanthreneslanthracenes 1.85 0.43 4 33.7 9.68 4 60.6 8.37 4 9.96 0.85 4 2.28 1.62 4 41.3 6.27 4
C4 henanthreneslanthracenes 1.05 0.23 4 19.2 6.36 4 66.9 15.5 4 6.61 0.23 4 0.71 0.48 4 13.0 2.51 4
fluoranthene 4.83 0.73 4 22.0 1.30 4 187 18.2 4 94.0 11.0 4 12.4 11.0 4 37.5 0.48 4
rene 4.25 0.67 4 20.6 1.21 4 169 14.5 4 83.3 9.54 4 9.73 8.70 4 33.9 0.55 4
Cl fluoranthenes/ renes 3.34 0.32 4 22.5 2.59 4 214 16.7 4 82.6 9.58 4 47.0 50.0 4 48.0 1.59 4
benz a anthracene 3.06 0.34 4 16.2 0.94 4 133 13.0 4 66.1 7.88 4 8.25 8.00 4 26.4 0.53 4
chr sene 6.56 1.54 4 20.3 0.86 4 161 13.9 4 75.8 9.26 4 10.3 8.90 4 33.3 0.72 4
Cl chr senes 7.90 1.96 4 42.9 2.32 4 319 23.1 4 119 12.7 4 23.1 19.7 4 72.0 3.10 4
C2 chr senes 4.67 1.66 4 33.7 5.40 4 212 24.0 4 47.3 7.51 4 11.0 8.89 4 43.8 4.61 4
C3 chr senes 4.89 2.53 4 27.3 3.73 4 178 34.3 4 24.0 2.23 4 ND ND ND 30.4 3.68 4
C4 chr senes ND ND ND ND ND ND ND ND ND ND ND ND ND ND ND ND ND ND
benzo b+k fluoranthene 9.18 1.59 4 33.6 1.98 4 253 14.6 4 123 14.5 4 16.9 15.8 4 50.8 1.15 4
benzo e rene 2.87 0.93 4 11.2 0.74 4 104 6.29 4 49.3 5.82 4 5.38 4.24 4 12.7 0.52 4
benzo a rene 5.60 0.46 4 20.4 1.34 4 158 8.05 4 68.1 7.33 4 12.1 13.4 4 35.2 1.49 4
er lene 2.74 0.39 4 8.84 0.46 4 52.4 3.78 4 23.9 2.24 4 3.74 3.82 4 10.7 0.26 4
indeno 1,2,3-cd rene 4.17 0.47 4 36.4 4.03 4 289 22.6 4 111 13.5 4 13.9 17.1 4 53.4 4.57 4
dibenz ah anthracene 3.23 0.33 4 9.96 1.33 4 65.1 5.96 4 25.2 3.27 4 4.14 4.73 4 14.7 1.67 4
benzo hi er lene 3.26 0.40 4 16.4 1.10 4 135 9.93 4 51.7 6.00 4 7.46 8.06 4 27.0 0.73 4

Total PAH16 (mg/kg) 47.6 5.88 4 208 13.7 4 1690 129 4 743 85.8 4 109 102 4 335 6.91 4
Total PAH34 (mg/kg) 82.7 14.7 4 451 46.7 4 3060 207 4 1150 129 4 219 199 4 661 16.2 4
Ratio PAH16/PAH34 0.58 0.03 4 0.46 0.03 4 0.55 0.02 4 0.65 0.00 4 0.48 0.03 4 0.51 0.02 4
Toxic Units (TU34) 1.7 0.3 4 6.2 0.6 4 8.9 0.6 4 4.5 0.5 4 2.6 2.3 4 6.7 0.1 4
ND - Non detected
Table 4-la
Page 1 of 2
TABLE 4-la
Sediment Contaminant Bioavailability Alliance
Total PAH Concentrations
Alcoa Badin Works Site
BL11 BL12 BL14 BL18 BL20 BL21
Sediment PAH's m /k Mean Std Dev. N Mean Std Dev. N Mean Std Dev. N Mean Std Dev. N Mean Std Dev. N Mean Std Dev. N
naphthalene 0.12 0.01 3 1.05 1.75 4 0.35 0.31 4 0.03 0.01 4 0.02 0.00 4 0.06 0.00 4
2-meth Ina hthalene 0.11 0.01 3 0.73 1.25 4 0.19 0.08 4 0.01 0.00 4 0.01 0.00 4 0.02 0.00 4
1-meth Ina hthalene 0.06 0.01 3 0.41 0.70 4 0.10 0.05 4 0.01 0.00 4 0.00 0.00 4 0.01 0.00 4
C2 naphthalenes 1.01 0.05 3 1.65 2.23 4 0.81 0.13 4 0.18 0.04 4 0.08 0.01 4 0.64 0.13 4
C3 naphthalenes 0.81 0.26 3 0.77 0.96 4 0.36 0.05 4 0.09 0.02 4 0.05 0.01 4 0.20 0.07 4
C4 naphthalenes 0.84 0.25 3 0.30 0.23 4 0.23 0.09 4 0.10 0.02 4 0.05 0.01 4 0.18 0.03 4
acena hth lene 0.09 0.03 3 0.11 0.08 4 0.16 0.01 4 0.02 0.01 4 0.01 0.01 4 0.13 0.11 4
acena hthene 0.32 0.01 3 5.45 10.2 4 0.25 0.21 4 0.03 0.03 4 0.01 0.00 4 0.02 0.01 4
fluorene 0.23 0.03 3 4.45 8.34 4 0.20 0.14 4 0.03 0.03 4 0.01 0.00 4 0.02 0.00 4
C1 fluorenes 1.17 0.02 3 2.04 3.29 4 0.41 0.08 4 0.13 0.01 4 0.09 0.01 4 0.24 0.00 4
C2 fluorenes 1.24 0.25 3 0.91 1.15 4 0.27 0.07 4 0.21 0.03 4 0.22 0.04 4 0.87 0.26 4
C3 fluorenes ND ND ND ND ND ND ND ND ND ND ND ND ND ND ND ND ND ND
henanthrene 2.89 0.10 3 30.4 54.1 4 2.12 1.21 4 0.28 0.27 4 0.05 0.01 4 0.11 0.01 4
anthracene 1.35 0.07 3 22.5 41.4 4 0.64 0.26 4 0.11 0.10 4 0.01 0.00 4 0.04 0.01 4
C1 henanthrenes/anthracenes 3.21 0.03 3 19.5 32.4 4 1.53 0.43 4 0.25 0.12 4 0.12 0.02 4 0.22 0.03 4
C2 henanthrenes/anthracenes 3.73 0.57 3 8.37 12.0 4 1.13 0.31 4 0.42 0.13 4 2.87 0.08 4 4.18 0.06 4
C3 henanthrenes/anthracenes 6.63 1.51 3 6.72 6.56 4 0.76 0.16 4 0.33 0.06 4 0.23 0.01 4 0.44 0.10 4
C4 henanthrenes/anthracenes 3.31 1.03 3 3.21 2.04 4 0.36 0.17 4 0.08 0.01 4 0.14 0.02 4 0.49 0.32 4
fluoranthene 8.40 0.17 3 42.0 67.7 4 3.71 1.45 4 0.55 0.45 4 0.03 0.01 4 0.06 0.01 4
rene 7.52 0.21 3 39.4 64.1 4 3.21 1.18 4 0.48 0.40 4 0.02 0.00 4 0.06 0.01 4
C1 fluoranthenesl renes 6.67 0.38 3 40.8 68.2 4 2.39 0.69 4 0.38 0.30 4 0.03 0.01 4 0.09 0.03 4
benz a anthracene 6.30 0.20 3 31.8 52.2 4 2.32 0.70 4 0.39 0.33 4 0.00 0.00 4 0.02 0.00 4
ch sene 8.52 0.21 3 42.3 67.7 4 3.15 0.79 4 0.53 0.39 4 0.01 0.00 4 0.04 0.01 4
C1 chr senes 15.2 1.35 3 41.5 57.3 4 4.51 1.12 4 0.76 0.63 4 0.02 0.00 4 0.10 0.00 4
C2 chr senes 10.8 1.54 3 20.4 18.7 4 2.18 0.53 4 0.40 0.32 4 ND ND ND 0.06 0.04 4
C3 chr senes 13.9 7.30 3 14.6 10.2 4 ND ND ND ND ND ND ND ND ND ND ND ND
C4 chr senes ND ND ND ND ND ND ND ND ND ND ND ND ND ND ND ND ND ND
benzo b+k fluoranthene 14.6 0.61 3 54.4 79.3 4 6.75 1.41 4 1.16 0.82 4 0.02 0.01 4 0.08 0.01 4
benzo e rene 5.25 0.36 3 12.2 16.0 4 2.92 0.55 4 0.51 0.34 4 0.01 0.00 4 0.03 0.01 4
benzo a rene 8.99 0.54 3 56.7 89.4 4 2.67 0.69 4 0.63 0.54 4 0.01 0.01 4 0.04 0.00 4
er lene 5.76 0.24 3 19.8 26.3 4 1.81 0.26 4 0.33 0.19 4 0.00 0.00 4 2.58 0.05 4
indeno 1,2,3-cd rene 11.1 1.24 3 96.2 165 4 2.68 0.70 4 0.48 0.39 4 0.01 0.00 4 0.04 0.01 4
dibenz ah anthracene 3.76 0.23 3 33.0 57.5 4 0.70 0.15 4 0.11 0.09 4 0.00 0.00 4 0.01 0.00 4
benzo hi er lene 5.82 0.32 3 30.9 50.2 4 2.19 0.53 4 0.42 0.33 4 0.01 0.00 4 0.03 0.00 4

Total PAH16 (mg/kg) 80.0 3.21 3 491 809 4 31.1 9.62 4 5.24 4.00 4 0.23 0.05 4 0.76 0.16 4
Total PAH34 (mg/kg) 160 6.27 3 685 1060 4 51.1 14.0 4 9.43 6.09 4 4.15 0.19 4 11.1 0.68 4
Ratio PAH16/PAH34 0.50 0.03 3 0.61 0.14 4 0.61 0.02 4 0.51 0.10 4 0.05 0.01 4 0.07 0.01 4
Toxic Units (TU34) 4.1 0.1 3 20.1 31.5 4 1.3 0.4 4 0.3 0.2 4 2.1 0.1 4 0.8 0.0 4
ND - Non detected
Table 4-la
Page 2 of 2
TABLE 4-lb
Sediment Contaminant Bioavailability Alliance
Porewater PAH Concentrations
Alcoa Badin Works Site
BL01 BL05 BL06 BL08 BL09 BL10
Sediment SPME Porewater PAHs (pg/L) Mean Std Dev N Mean Std Dev N Mean Std Dev N Mean Std Dev N Mean Std Dev N Mean Std Dev N
naphthalene 0.096 0.045 4 ND ND ND ND ND ND ND ND ND ND ND ND 0.084 0.049 4
2-meth Ina hthalene 0.014 0.003 4 0.043 0.001 4 0.031 0.003 4 0.009 0.002 4 ND ND ND 0.015 0.007 4
1-meth Ina hthalene 0.012 0.003 4 0.415 0.008 4 0.260 0.007 4 0.014 0.002 4 ND ND ND 0.016 0.010 4
C2 naphthalenes ND ND ND 20.48 1.189 4 8.380 0.647 4 0.654 0.153 4 ND ND ND 2.927 0.325 4
C3 naphthalenes ND ND ND 28.08 1.794 4 10.60 1.158 4 0.244 0.010 4 ND ND ND 1.919 0.342 4
C4 naphthalenes ND ND ND 12.18 1.016 4 6.177 0.949 4 ND ND ND ND ND ND 1.586 0.335 4
acena hth lene 0.006 0.003 4 0.145 0.012 4 0.075 0.006 4 0.008 0.001 4 ND ND ND 0.023 0.003 4
acena hthene 0.057 0.002 4 0.340 0.007 4 0.342 0.017 4 0.276 0.016 4 ND ND ND 0.201 0.029 4
fluorene 0.014 0.004 4 0.145 0.008 4 0.122 0.008 4 0.039 0.005 4 ND ND ND 0.027 0.004 4
C1 fluorenes ND ND ND 1.469 0.031 4 0.653 0.050 4 0.128 0.012 4 ND ND ND 0.248 0.031 4
C2fluorenes ND ND ND 3.617 0.345 4 1.871 0.248 4 ND ND ND ND ND ND 0.895 0.037 4
C3fluorenes ND ND ND ND ND ND ND ND ND ND ND ND ND ND ND ND ND ND
henanthrene 0.031 0.002 4 0.329 0.004 4 0.531 0.018 4 0.119 0.008 4 ND ND ND 0.119 0.021 4
anthracene 0.010 0.001 4 0.099 0.003 4 0.128 0.007 4 0.045 0.002 4 ND ND ND 0.058 0.011 4
C1 henanthrenes/anthracenes ND ND ND 2.012 0.047 4 1.246 0.063 4 0.139 0.014 4 ND ND ND 0.212 0.034 4
C2 henanthrenes/anthracenes ND ND ND 3.376 0.112 4 1.798 0.200 4 0.103 0.012 4 ND ND ND 0.376 0.049 4
C3 henanthrenes/anthracenes ND ND ND 1.841 0.094 4 1.135 0.159 4 ND ND ND ND ND ND 0.374 0.067 4
C4 henanthrenes/anthracenes ND ND ND 0.640 0.072 4 0.394 0.069 4 ND ND ND ND ND ND 0.393 0.044 4
fluoranthene 0.064 0.004 4 0.274 0.003 4 0.501 0.015 4 0.510 0.049 4 ND ND ND 0.305 0.053 4
rene 0.066 0.002 4 0.311 0.004 4 0.457 0.012 4 0.426 0.035 4 ND ND ND 0.293 0.051 4
C1 fluoranthenes/ renes ND ND ND 0.774 0.022 4 0.783 0.024 4 0.595 0.018 4 ND ND ND 0.561 0.039 4
benz a anthracene 0.003 0.001 4 0.019 0.001 4 0.036 0.002 4 0.031 0.004 4 ND ND ND 0.016 0.005 4
ch sene 0.013 0.001 4 0.037 0.001 4 0.067 0.002 4 0.055 0.005 4 ND ND ND 0.035 0.008 4
C1 chr senes ND ND ND 0.034 0.001 4 ND ND ND ND ND ND ND ND ND 0.021 0.007 4
C2 chr senes ND ND ND ND ND ND ND ND ND ND ND ND ND ND ND ND ND ND
C3 chr senes ND ND ND ND ND ND ND ND ND ND ND ND ND ND ND ND ND ND
C4 chr senes ND ND ND ND ND ND ND ND ND ND ND ND ND ND ND ND ND ND
benzob+kfluoranthene ND ND ND 0.006 0.002 4 0.014 0.002 4 0.021 0.003 4 ND ND ND 0.007 0.002 4
benzo a rene ND ND ND 0.008 0.002 4 0.020 0.004 4 0.015 0.002 4 ND ND ND 0.007 0.002 4
benzo a rene ND ND ND ND ND ND ND ND ND 0.012 0.002 4 ND ND ND ND ND ND
er lene ND ND ND ND ND ND ND ND ND ND ND ND ND ND ND ND ND ND
indeno 1 2 3-cd rene ND ND ND ND ND ND ND ND ND ND ND ND ND ND ND ND ND ND
dibenz ah anthracene ND ND ND ND ND ND ND ND ND ND ND ND ND ND ND ND ND ND
benzo hi a lene ND ND ND ND ND ND ND ND ND ND ND ND ND ND ND ND ND ND

SPME PAH16 (mg/kg) 0.35 0.05 4 1.71 0.01 4 2.27 0.06 4 1.543 0.124 4 ND ND ND 1.17 0.15 4
SPME PAH34 (mg/kg) 0.38 0.05 4 76.7 4.51 4 35.6 3.22 4 3.445 0.280 4 ND ND ND 10.7 1.32 4

SPME Toxic Units (TU16) 0.02 0.00 4 0.14 0.00 4 0.23 0.00 4 0.214 0.019 4 ND ND ND 0.12 0.02 4
SPME Toxic Units (TU34) 0.03 0.00 4 11.29 0.70 4 5.91 0.61 4 0.457 0.029 4 ND ND ND 2.26 0.27 4
SPME TU16/SPME TU34 0.99 0.00 4 0.01 0.00 4 0.04 0.00 4 0.469 0.013 4 ND ND ND 0.05 0.01 4
ND - Non detected
Table 4-lb
Page 1 of 2
TABLE 4-lb
Sediment Contaminant Bioavailability Alliance
Porewater PAH Concentrations
Alcoa Badin Works Site
BL11 BL12 BL14 BL18 BL20 BL21
Sediment SPME Porewater PAHs (pg/L) Mean Std Dev N Mean Std Dev N Mean Std Dev N Mean Std Dev N Mean Std Dev N Mean Std Dev N
naphthalene ND ND ND ND ND ND ND ND ND ND ND ND ND ND ND ND ND ND
2-meth Ina hthalene 0.011 0.003 4 ND ND ND ND ND ND ND ND ND ND ND ND ND ND ND
1-meth Ina hthalene 0.014 0.005 4 ND ND ND ND ND ND ND ND ND 0.015 0.003 4 ND ND ND
C2 naphthalenes 0.702 0.117 4 ND ND ND ND ND ND ND ND ND ND ND ND ND ND ND
C3 naphthalenes 2.650 0.097 4 0.115 0.021 4 ND ND ND ND ND ND ND ND ND ND ND ND
C4 naphthalenes 2.211 0.114 4 0.230 0.056 4 ND ND ND ND ND ND ND ND ND ND ND ND
acena hth lene 0.017 0.006 4 ND ND ND ND ND ND ND ND ND 0.011 0.002 4 ND ND ND
acena hthene 0.083 0.009 4 0.010 0.003 4 ND ND ND 0.008 0.003 4 0.029 0.008 4 0.032 0.006 4
fluorene 0.008 0.003 4 ND ND ND ND ND ND 0.012 0.004 4 0.018 0.006 4 ND ND ND
C1 fluorenes 0.246 0.031 4 0.059 0.013 4 ND ND ND ND ND ND ND ND ND ND ND ND
C2fluorenes 0.988 0.097 4 ND ND ND ND ND ND ND ND ND ND ND ND ND ND ND
C3fluorenes ND ND ND ND ND ND ND ND ND ND ND ND ND ND ND ND ND ND
henanthrene 0.103 0.007 4 0.009 0.002 4 ND ND ND ND ND ND ND ND ND ND ND ND
anthracene 0.040 0.004 4 0.004 0.001 4 ND ND ND ND ND ND ND ND ND 0.001 0.000 4
C1 henanthrenes/anthracenes 0.425 0.041 4 ND ND ND ND ND ND ND ND ND ND ND ND ND ND ND
C2 henanthrenes/anthracenes 0.709 0.082 4 ND ND ND ND ND ND ND ND ND ND ND ND ND ND ND
C3 henanthrenes/anthracenes 0.434 0.022 4 ND ND ND ND ND ND ND ND ND ND ND ND ND ND ND
C4 henanthrenes/anthracenes 0.178 0.037 4 ND ND ND ND ND ND ND ND ND ND ND ND ND ND ND
fluoranthene 0.255 0.024 4 0.060 0.002 4 0.012 0.001 4 0.002 0.001 4 0.004 0.002 4 ND ND ND
rene 0.240 0.020 4 0.069 0.003 4 0.011 0.001 4 0.001 0.001 4 0.003 0.002 4 ND ND ND
C1 fluoranthenes/ renes 0.589 0.029 4 0.048 0.004 4 ND ND ND ND ND ND ND ND ND ND ND ND
benz a anthracene 0.015 0.003 4 0.004 0.000 4 ND ND ND ND ND ND ND ND ND ND ND ND
ch sene 0.036 0.004 4 0.011 0.000 4 ND ND ND 0.000 0.000 4 ND ND ND ND ND ND
C1 chr senes ND ND ND ND ND ND ND ND ND ND ND ND ND ND ND ND ND ND
C2 chr senes ND ND ND ND ND ND ND ND ND ND ND ND ND ND ND ND ND ND
C3 chr senes ND ND ND ND ND ND ND ND ND ND ND ND ND ND ND ND ND ND
C4 chr senes ND ND ND ND ND ND ND ND ND ND ND ND ND ND ND ND ND ND
benzob+kfluoranthene 0.010 0.001 4 ND ND ND ND ND ND ND ND ND ND ND ND ND ND ND
benzo a rene 0.009 0.002 4 ND ND ND ND ND ND ND ND ND ND ND ND ND ND ND
benzo a rene ND ND ND ND ND ND ND ND ND ND ND ND ND ND ND ND ND ND
er lene ND ND ND ND ND ND ND ND ND ND ND ND ND ND ND ND ND ND
indeno 1 2 3-cd rene ND ND ND ND ND ND ND ND ND ND ND ND ND ND ND ND ND ND
dibenz ah anthracene ND ND ND ND ND ND ND ND ND ND ND ND ND ND ND ND ND ND
benzo hi a lene ND ND ND ND ND ND ND ND ND ND ND ND ND ND ND ND ND ND

SPME PAH16 (mg/kg) 0.81 0.07 4 0.17 0.01 4 0.02 0.00 4 0.02 0.01 4 0.07 0.02 4 0.03 0.01 4
SPME PAH34 (mg/kg) 9.97 0.57 4 0.62 0.07 4 0.02 0.00 4 0.02 0.01 4 0.08 0.02 4 0.03 0.01 4

SPME Toxic Units (TU16) 0.11 0.01 4 0.02 0.00 4 0.00 0.00 4 0.00 0.00 4 0.00 0.00 4 0.00 0.00 4
SPME Toxic Units (TU34) 2.19 0.10 4 0.10 0.02 4 0.00 0.00 4 0.00 0.00 4 0.00 0.00 4 0.00 0.00 4
SPME TU16/SPME TU34 0.05 0.00 4 0.23 0.04 4 1.00 0.00 4 1.00 0.00 4 0.90 0.03 4 1.00 0.00 4
ND - Non detected
Table 4-lb
Page 2 of 2
TABLE 4-lc
Sediment Contaminant Bioavailability Alliance
SFE PAH Concentrations
Alcoa Badin Works Site
BL01 BL05 BL06 BL08 BL09 BL10
Sediment SFE P! H m /k Mean Std Dev N Mean Std Dev N Mean Std Dev N Mean Std Dev N Mean Std Dev N Mean Std Dev N
naphthalene 0.007 0.002 4 0.005 0.002 4 0.031 0.023 4 0.008 0.005 4 0.007 0.002 4 0.009 0.002 4
2-methyl nahthalene 0.003 0.001 4 0.002 0.001 4 0.026 0.013 4 0.004 0.000 4 0.003 0.001 4 0.004 0.002 4
1-meth Inaphthalene 0.003 0.001 4 0.004 0.002 4 0.031 0.014 4 0.003 0.001 4 0.002 0.001 4 0.003 0.001 4
C2 naphthalenes 0.052 0.001 4 0.213 0.119 4 0.719 0.404 4 0.037 0.005 4 ND ND ND 0.086 0.028 4
C3 naphthalenes 0.016 0.003 4 0.407 0.222 4 0.873 0.588 4 0.019 0.004 4 ND ND ND 0.127 0.054 4
C4 naphthalenes 0.010 0.003 4 0.242 0.146 4 0.383 0.264 4 ND ND ND ND ND ND 0.072 0.048 4
acena hth lene 0.002 0.000 4 0.001 0.001 4 0.006 0.002 4 0.002 0.001 4 0.001 0.000 4 0.003 0.002 4
acena hthene 0.004 0.001 4 0.006 0.003 4 0.084 0.039 4 0.024 0.014 4 0.004 0.001 4 0.013 0.004 4
fluorene 0.004 0.001 4 0.005 0.003 4 0.056 0.031 4 0.017 0.009 4 0.003 0.001 4 0.008 0.002 4
C1 fluorenes 0.009 0.005 4 0.072 0.046 4 0.193 0.109 4 0.020 0.009 4 ND ND ND 0.032 0.019 4
C2 fluorenes 0.019 0.009 4 0.249 0.158 4 0.375 0.237 4 ND ND ND ND ND ND 0.066 0.042 4
C3 fluorenes ND ND ND ND ND ND ND ND ND ND ND ND ND ND ND ND ND ND
henanthrene 0.026 0.012 4 0.033 0.022 4 0.465 0.247 4 0.181 0.117 4 0.024 0.011 4 0.065 0.032 4
anthracene 0.010 0.002 4 0.012 0.008 4 0.179 0.091 4 0.059 0.040 4 0.008 0.004 4 0.030 0.013 4
C1 henanthreneslanthracenes 0.041 0.009 4 0.310 0.190 4 0.792 0.513 4 0.088 0.049 4 0.021 0.009 4 0.099 0.059 4
C2 henanthreneslanthracenes 0.026 0.010 4 2.053 1.327 4 2.084 1.422 4 0.090 0.041 4 0.019 0.007 4 0.999 0.741 4
C3 henanthreneslanthracenes 0.034 0.013 4 3.453 2.383 4 3.457 2.086 4 0.066 0.036 4 ND ND ND 2.189 1.414 4
C4 henanthreneslanthracenes 0.057 0.035 4 1.939 1.388 4 4.267 2.300 4 0.062 0.026 4 0.001 0.000 3 0.872 0.450 4
fluoranthene 0.039 0.019 4 0.092 0.066 4 0.924 0.425 4 0.467 0.297 4 0.024 0.013 4 0.190 0.081 4
rene 0.037 0.017 4 0.114 0.078 4 0.815 0.374 4 0.374 0.240 4 0.020 0.010 4 0.165 0.067 4
C1 fluoranthenes/ renes 0.022 0.007 4 0.374 0.260 4 1.377 0.694 4 0.282 0.192 4 0.058 0.034 4 0.228 0.105 4
benz a anthracene 0.025 0.019 4 0.058 0.041 4 0.442 0.200 4 0.221 0.152 4 0.013 0.007 4 0.102 0.029 4
chr sene 0.080 0.049 4 0.114 0.078 4 0.662 0.290 4 0.284 0.189 4 0.023 0.013 4 0.189 0.071 4
C1 chr senes 0.127 0.034 4 0.751 0.522 4 1.974 0.943 4 0.299 0.190 4 0.038 0.022 4 0.575 0.213 4
C2 chr senes ND ND ND 1.398 1.086 4 3.395 1.633 4 ND ND ND ND ND ND 0.726 0.327 4
C3 chr senes ND ND ND 1.525 1.145 4 4.425 2.072 4 ND ND ND ND ND ND ND ND ND
C4 ch senes ND ND ND ND ND ND ND ND ND ND ND ND ND ND ND ND ND ND
benzo b+k fluoranthene 0.059 0.041 4 0.085 0.056 4 0.823 0.332 4 0.341 0.201 4 0.028 0.018 4 0.186 0.053 4
benzo e rene 0.026 0.017 4 0.043 0.027 4 0.349 0.144 4 0.110 0.067 4 0.011 0.006 4 0.071 0.018 4
benzo a rene 0.037 0.026 4 0.040 0.024 4 0.357 0.147 4 0.128 0.081 4 0.029 0.012 4 0.087 0.022 4
er lene 0.023 0.014 4 0.022 0.013 4 0.119 0.052 4 0.037 0.023 4 0.009 0.007 4 0.039 0.010 4
indeno 1,2,3-cd rene ND ND ND 0.015 0.013 4 0.378 0.168 4 0.083 0.062 4 0.008 0.006 4 0.043 0.010 4
dibenz ah anthracene ND ND ND 0.013 0.008 4 0.093 0.039 4 0.019 0.012 4 0.007 0.005 3 0.034 0.006 4
benzo hi e lene ND ND ND 0.017 0.010 4 0.170 0.072 4 0.043 0.027 4 0.005 0.003 4 0.033 0.007 4

SFE PAH16 (mg/kg) 0.33 0.18 4 0.61 0.41 4 5.48 2.43 4 2.25 1.44 4 0.20 0.10 4 1.16 0.38 4
SFE PAH34 (mg/kg) 0.80 0.23 4 13.66 9.38 4 30.3 15.6 4 3.37 2.07 4 0.37 0.18 4 7.35 3.67 4

SFE Toxic Units (TU16) 0.01 0.00 4 0.01 0.01 4 0.02 0.01 4 0.01 0.01 4 0.00 0.00 4 0.01 0.00 4
SFE Toxic Units (TU34) 0.02 0.00 4 0.18 0.12 4 0.09 0.04 4 0.01 0.01 4 0.00 0.00 4 0.07 0.04 4
SFE TU161SFE TU34 0.39 0.09 4 0.05 0.01 4 0.21 0.04 4 0.65 0.07 4 0.56 0.02 4 0.19 0.07 4
ND - Non detected
Table 4-lc
Page 1 of 2
TABLE 4-lc
Sediment Contaminant Bioavailability Alliance
SFE PAH Concentrations
Alcoa Badin Works Site
BL11 BL12 BL14 BL18 BL20 BL21
Sediment SFE P! H m Ik Mean Std Dev N Mean Std Dev N Mean Std Dev N Mean Std Dev N Mean Std Dev N Mean Std Dev N
naphthalene 0.003 0.001 3 0.015 0.020 4 0.004 0.002 4 0.003 0.001 4 0.003 0.001 3 0.004 0.001 4
2-methyl nahthalene 0.002 0.000 3 0.010 0.015 4 0.002 0.001 4 0.002 0.000 4 0.001 0.000 3 0.002 0.000 4
1-meth Inaphthalene 0.002 0.000 3 0.006 0.009 4 0.001 0.001 4 0.002 0.000 4 0.001 0.001 3 0.001 0.001 4
C2 naphthalenes 0.055 0.005 3 0.163 1 0.041 0.009 4 0.024 0.002 4 0.015 0.004 3 0.023 0.003 4
C3 naphthalenes 0.102 0.056 3 0.074 1 0.007 0.003 4 0.017 0.002 4 0.008 0.004 3 0.013 0.002 4
C4 naphthalenes 0.130 0.030 3 0.025 1 0.004 0.004 3 0.007 0.001 4 0.003 0.002 3 0.007 0.001 4
acena hth lene 0.005 0.005 3 0.002 0.002 4 0.001 0.000 4 0.001 0.000 4 0.001 0.000 3 0.001 0.001 4
acena hthene 0.007 0.004 3 0.023 0.039 4 0.002 0.001 4 0.002 0.000 4 0.002 0.001 3 0.003 0.000 4
fluorene 0.004 0.002 3 0.019 0.033 4 0.002 0.000 4 0.003 0.001 4 0.004 0.002 3 0.004 0.001 4
C1 fluorenes 0.204 0.186 3 0.032 1 ND ND ND 0.007 0.001 4 0.005 0.002 3 0.008 0.002 4
C2 fluorenes 0.146 0.086 3 0.030 1 0.005 0.001 4 0.010 0.003 4 0.006 0.002 3 0.013 0.003 4
C3 fluorenes ND ND ND ND ND ND ND ND ND ND ND ND ND ND ND ND ND ND
henanthrene 0.034 0.022 3 0.110 0.182 4 0.011 0.005 4 0.012 0.003 4 0.016 0.009 3 0.019 0.004 4
anthracene 0.019 0.007 3 0.054 0.090 4 0.003 0.001 4 0.003 0.001 4 0.004 0.003 3 0.005 0.001 4
C1 henanthreneslanthracenes 0.173 0.061 3 0.073 0.097 4 0.009 0.004 4 0.010 0.003 4 0.010 0.006 3 0.013 0.001 4
C2 henanthreneslanthracenes 0.340 0.206 3 0.055 0.060 4 0.007 0.004 4 0.014 0.003 4 1.255 0.025 3 1.843 0.028 4
C3 henanthreneslanthracenes 0.860 0.467 3 0.187 0.155 4 ND ND ND 0.009 0.002 4 0.019 0.003 3 0.017 0.005 4
C4 henanthreneslanthracenes 0.503 0.451 3 0.217 0.160 4 ND ND ND 0.011 0.004 4 0.010 0.001 3 0.022 0.004 4
fluoranthene 0.103 0.085 3 0.107 0.151 4 0.014 0.009 4 0.009 0.002 4 0.010 0.004 3 0.014 0.004 4
rene 0.094 0.072 3 0.092 0.121 4 0.011 0.007 4 0.008 0.001 4 0.008 0.001 3 0.012 0.003 4
C1 fluoranthenes/ renes 0.144 0.110 3 0.063 0.074 4 0.009 0.004 4 0.004 0.001 4 0.005 0.002 3 0.007 0.001 4
benz a anthracene 0.070 0.065 3 0.059 0.079 4 0.005 0.004 4 0.002 0.001 4 0.002 0.001 3 0.003 0.001 4
chr sene 0.173 0.165 3 0.117 0.153 4 0.009 0.006 4 0.004 0.001 4 0.003 0.001 3 0.005 0.001 4
C1 chr senes 0.499 0.232 3 0.258 0.240 4 ND ND ND 0.004 0.001 4 0.005 0.000 3 0.008 0.001 4
C2 chr senes 0.876 0.785 3 0.762 1 ND ND ND 0.002 0.001 3 ND ND ND 0.009 0.002 4
C3 chr senes 1.070 0.792 3 1.056 1 ND ND ND ND ND ND ND ND ND ND ND ND
C4 ch senes ND ND ND ND ND ND ND ND ND ND ND ND ND ND ND ND ND ND
benzo b+k fluoranthene 0.134 0.095 3 0.141 0.129 4 0.014 0.010 4 0.005 0.001 4 0.003 0.000 3 0.007 0.001 4
benzo e rene 0.059 0.043 3 0.050 0.040 4 0.005 0.004 4 0.002 0.001 4 0.001 0.000 3 0.002 0.000 4
benzo a rene 0.053 0.028 3 0.081 0.060 4 0.005 0.003 4 ND ND ND 0.002 0.000 3 0.004 0.001 4
er lene 0.049 0.028 3 0.059 0.036 4 0.006 0.005 4 0.002 0.000 4 0.001 0.000 3 0.013 0.004 4
indeno 1,2,3-cd rene 0.024 0.014 3 0.021 0.019 4 0.003 0.002 4 0.001 0.000 4 0.002 0.001 3 0.003 0.000 4
dibenz ah anthracene ND ND ND ND ND ND ND ND ND 0.000 0.000 4 0.000 0.000 3 0.001 0.000 4
benzo hi a lene 0.017 0.011 3 0.015 0.012 4 0.003 0.003 4 0.002 0.000 4 0.002 0.001 3 0.003 0.000 4

SFE PAH16 (mg/Kg) 0.74 0.56 3 0.86 1.08 4 0.09 0.05 4 0.05 0.01 4 0.06 0.02 3 0.09 0.02 4
SFE PAH34 (mg/Kg) 5.95 3.59 3 2.37 3.01 4 0.18 0.07 4 0.18 0.02 4 1.41 0.07 3 2.09 0.04 4

SFE Toxic Units (TU16) 0.02 0.01 3 0.03 0.04 4 0.00 0.00 4 0.00 0.00 4 0.04 0.01 3 0.01 0.00 4
SFE Toxic Units (TU34) 0.15 0.08 3 0.07 0.09 4 0.01 0.00 4 0.01 0.00 4 0.71 0.04 3 0.12 0.00 4
SFE TU161SFE TU34 0.13 0.03 3 0.38 0.02 4 0.40 0.10 4 0.30 0.05 4 0.05 0.01 3 0.05 0.01 4
ND - Non detected
Table 4-lc
Page 2 of 2
TABLE 4-2
Sediment Contaminant Bioavailability Assessment
Chemical and Physical Characteristics
Alcoa Badin Works Site
Sample BI-01 BI-06 BI-06 BI-08 BI-09 BL10 BL11 BL12 BL14 BL18 BL20 BL21
Grain Size Data
Sand (%) 44.7 6.4 54.0 35.8 77.1 8.2 16.2 21.0 33.0 50.8 88.3 1.1
Silt (%) 27.7 29.8 18.6 36.0 14.3 37.4 33.8 27.3 40.4 26.6 10.2 36.7
Clay (%) 27.6 63.8 27.4 28.3 8.6 54.4 50.0 51.7 26.6 22.6 1.6 62.2
Percent Solids
Solids (%) 51.8 35.3 46.4 48 71.2 38.7 44.9 45.6 51.5 47.1 70.3 26.2
Organic Carbon
DOC (mg/L) 6.30 2.00 2.40 1.90 2.20 2.70 3.00 2.70 2.10 2.40 1.90 2.60
Sediment TOC (wt. %) 6.00 8.97 42.4 32.2 10.9 12.2 4.83 4.42 4.99 3.65 0.27 2.34
Sediment Soot C (wt. %) 5.22 7.29 39.7 32.1 12.0 10.3 3.30 2.85 2.61 0.65 0.07 0.20
Basic Chemistry q
Ammonia (mg/Kg) 34.1 64.7 58 56 13.0 67 103 58 46 93 10 109
pH 6.9 6.8 6.9 6.7 6.8 6.8 6.9 6.8 6.8 6.9 6.9 6.5
NA - Not analyzed
Table 4-2
TABLE 4-3
Sediment Contaminant Bioavailability Assessment
Cyanide Analysis
Alcoa Badin Works Site
Sample ID Total Cyanide*
m k Free Cyanide**
(mg/kg) Iron Cyanide***
m k
BLO1 0.66 0.10 0.36 J/UJ
BL05 2.12 0.22 1.49 J/UJ
BL06 1.75 0.13 1.09 J/UJ
BLO8 0.66 0.10 0.41 J/UJ
BL09 4.53 0.04 2.88 J/UJ
BL10 6.66 0.20 3.86 J/UJ
BL11 3.67 0.14 2.19 J/UJ
BL12 1.09 0.16 0.69 J/UJ
BL14 3.59 0.15 1.04 J/UJ
BL18 0.17 0.09 <0.006 J/UJ
BL20 <0.05 <0.041 0.02 J/UJ
BL21 0.62 0.50 0.49 J/UJ
*Total Cyanide determined by APHA Standard Methods 4500-CV-C. "Total Cyanide
after Distillation" and APHA Standard Methods 4500-CV-E., "Colorimetric Method."
**Free Cyanide determined by ASTMD4282-95 . "Standard Test Method for
Determination of Free Cyanide in Water and Wastewater by Microdiffusion.
***Iron Cyanide determined by SW-846 Method 9015. "Metal Cyanide Complexes by
Anion Exchange Chromatography and UV Detection."
JIUJ values are estimates because of imprecision due to significant matrix interference.
Table 4-3
TABLE 4-4a
Sediment Contaminant Bioavailability Assessment
H. azteca Toxicity Test Data - Survival and Growth
Alcoa Badin Works Site
S
l
T Sample Survival Growth
amp
e
ype ID N (mg drywt.lorganism)

Means Standard
Deviationb
n°
Mean Standard
Deviation
n
Lab Control 95.0 8.4 6 0.19 0.04 6
Positive Control DNAPL 93.3 12.1 6 0.26 0.09 6
Positive Control Pitch 6.7e'' 16.3 6 0.26' NA 1
Test B1_01 96.7 8.2 6 0.21 0.04 6
Test B1_05 81.7 11.7 6 0.21 0.04 6
Test B1_06 75.0 18.7 6 0.21 0.06 6
Test B1_08 76.7 16.3 6 0.20 0.08 6
Test B1_09 93.3 8.2 6 0.13' 0.02 6
Test B1_10 90.0 12.6 6 0.15 0.06 6
Test B1_11 96.7 5.2 6 0.18 0.06 6
Test BL12 93.3 5.2 6 0.19 0.08 6
Test B1_14 86.7 20.7 6 0.19 0.04 6
Field Reference BL18 85.0 10.5 6 0.17 0.04 6
Field Coarse-grain Reference BL20 88.3 7.5 6 0.24 0.03 6
Field Reference BL21 88.3 20.4 6 0.24 0.07 6
aArithmetic mean
nStandard deviation
`Number of replicate analyses
a Excluded from the Tukey method of multiple comparisons for growth due to significant mortality in the replicate.
a Significantly different from BL18 reference
(Significantly different from BL21 reference
Comparisons of survival among treatments were conducted with arc sine (square root) transformed values subjected to either the parametric
Dunnett's Test or non-parametric Steel's Many-One Rank Test procedures. Dry weight among treatments was analyzed using the Tukey Method of
Multiple Comparisons.
Table 4-4a
TABLE 4-4b
Sediment Contaminant Bioavailability Assessment
C. tentans Toxicity Test Data - Survival and Growth
Alcoa Badin Works Site
S
l
T Sample Survival Growth
amp
e
ype ID N (mg drywt.lorganism)
Mean' Standard
Deviation Nc Mean Standard
Deviation N
Lab Control 92.9 11.1 7 2.3 0.3 7
Positive Control DNAPL 61.4e'' 10.7 7 2.6 0.6 7
Positive Control Pitch 60.0e' 8.2 7 1.8 0.3 7
Test BL01 94.3 7.9 7 2.0 0.3 7
Test BL05 95.7 7.9 7 2.4 0.7 7
Test BL06 84.3 19.0 7 2.4 0.6 7
Test BL08 94.3 9.8 7 2.2 0.4 7
Test BL09 82.9 17.0 7 2.3 0.4 7
Test BL10 77.1 26.3 7 2.7 0.8 7
Test BL11 62.9' 28.1 7 2.8 0.6 7
Test BL12 87.1 9.5 7 2.5 0.5 7
Test BL14 84.3 22.3 7 2.5 0.5 7
Field Reference BL18 88.6 10.7 7 2.3 0.4 7
Field Coarse-grain Reference BL20 87.1 19.8 7 2.1 0.7 7
Field Reference BL21 100.0 0.0 7 2.1 0.4 7
'Arithmetic mean
nStandard deviation
`Number of replicate analyses
aNo growth data collected in test sample due to 100% mortality
eSignificantly different from BL18 reference
(Significantly different from BL21 reference
Comparisons of survival among treatments were conducted with arc sine (square root) transformed values subjected to either the parametric
Dunnett's Test or non-parametric Steel's Many-One Rank Test procedures. Dry weight among treatments was analyzed using the Tukey Method of
Multiple Comparisons.
Table 4-4b
TABLE 4-5
Sediment Contaminant Bioavailability Assessment
Spiked Pitch and DNAPL Total PAH Concentrations
Alcoa Badin Works Site
Sediment PAH (mg/kg) Pitch NAPL
naphthalene 0.06 13.03
2-meth Ina hthalene 0.06 6.22
1-meth Ina hthalene 0.05 3.91
C2 naphthalenes 0.32 6.76
C3 naphthalenes 0.29 2.28
C4 naphthalenes 0.14 0.71
acena hth lene 0.02 1.00
acena hthene 0.73 3.04
fluorene 0.44 1.77
C1 fluorenes 0.40 2.49
C2 fluorenes 0.28 1.17
C3 fluorenes ND ND
henanthrene 4.75 7.80
anthracene 1.14 3.08
C1 henanthrenes/anthracenes 2.98 10.42
C2 henanthrenes/anthracenes 4.37 11.08
C3 henanthrenes/anthracenes 1.56 3.20
C4 henanthrenes/anthracenes 0.59 0.78
fluoranthene 6.61 2.38
rene 7.16 4.79
C1 fluoranthenes/ renes 8.75 7.55
benz a anthracene 5.93 2.12
ch sene 6.16 1.61
C1 ch senes 14.17 6.36
C2 ch senes 9.59 2.88
C3 ch senes 3.23 0.69
C4 ch senes ND ND
benzo b+k fluoranthene 8.45 1.25
benzo a rene 2.71 0.57
benzo a rene 6.13 1.45
e lene 1.40 0.24
indeno 1,2,3-cd rene 4.66 0.47
dibenz ah anthracene 1.32 0.16
benzo hi a lene 2.03 0.28

Total PAH16 (mg/kg) 55.6 44.3
Total PAH34 (mg/kg) 106.4 111.6
ND - Non detected
Table 4-5
Figures
'�t
9
I DATE: 1/04/05

Figure 4-1 Relative distribution of total extractable PAHs in Badin Lake, tar-, and pitch-
spiked control sediments
Q
a
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PAH
Figure 4-2 PAH concentrations compared to survival of H. azteca and C. tentans in
Badin Works sediment compared to other aluminum industry and MGP sites.
120
100
80
60
3
40
20
A. Total Extractable PAH16
----- - - - -- 0 - ------------
0 0 0
0
0
--------------------- --- a -- ---------------
0
----------------- -----------------------
0 4-
0.1
120
100
80
0
>-60
3
40
20
120
100
80
0
> 60
3
40
20
1 10 100 1000 10000 100000
Total PAH16 (mg/kg)
B. Dissolved PAH34 in Sediment Pore Water
---- -- -o-CY-- 0010$ 0 -0-131 ---------
0 I I
---- - -------01 --- -0 ------
0 -----------
I
I
I I
-------------------------------
01 0 1
-------------------------------F-
0 ----
I
a.?
0.0001 0.001 0.01 0.1 1 10 100
SPME Pore Water PAH34 (TU34)
-Probit regression - -85% Survival (6.7 TUs) - -15% Survival (32 TUs)
1000
C. Rapidly Released PAH34 in Sediment Determined By SFE
190 11 - o --ol
? I I
---- ------- - -- o---------i-------------
0
I I
-0 -J--- -?----I-------------
10 I
-------------------- ----
001 1
---------------------A-9------- - i -------------
I
0
0.001 0.01 0.1 1 10 100 1000
SFE Rapidly Released PAH34 (TU34)
-Probit regression - -85% Survival (0.6 TUs) - -15% Survival (19 TUs)
O Other MGP & Aluminum Industry Sites
* Badin Works H. azteca
? Badin Works C. tentans
Figure 4-3 PAH concentrations compared to growth of H. azteca and C. tentans
expressed as a percent change in weight compared to laboratory control sediment
160
140
120
t 100
U
s 80
a) 60
40
20
0

--1----1------1------- -1 711
----------
0.1 1 10 100 1000
Sediment Total PAH16 (mg/kg)
IOU
140
120
t 100
U
s 80
27,
a) 60
40
20
0
10000

---------- ----------
0.0001 0.001 0.01 0.1 1 10 100
Porewater PAH34 Concentration (SPME TU34)
IOU
140
120
a?
100
s
U 80
r
M
P 60
40
0
20
0
0.001 0.01 0.1 1 10
Rapidly Released PAH34 Concentration (SFE TU34)
0 Badin Lake - H. azteca 0 Badin Lake - C. tentans
Figure 4-4 Relationship between total organic carbon content and heat stable carbon
content in Badin Lake sediments
50
40
r-
0
30
U
m
M 20
4.0
Cn
c?
= 10
0

r2 = 0.99
0 10 20 30 40 50
Total Organic Carbon (%)
Figure 4-5 Relationship between total extractable PAH16 concentration and total organic
carbon content in Badin Lake sediments
50
0

c 40
0
L
v 30
20
L
0
4 10
0
?
r2 = 0.85

0
0 500 1000 1500 2000
Sediment Total PAH16 Concentration (mg/kg)
Appendix A
Digital Laboratory Data
(CD-ROM)
Appendix B
U.S. Army Corps ERDC Acute and Chronic Sediment
Toxicity Final Report for Badin Lake Sediment Exposed
Chironomus tentans and Hyalella azteca
CERDC-EP-R
23 September 2005
MEMORANDUM FOR: Mr. Joe Kreitinger/RETEC
SUBJECT: Acute and Chronic Sediment Toxicity Data for Badin Lake Sediment Exposed
Chironomus tentans and Hyalella azteca
The following is an abbreviated summary of the results from the 10-d Chironomus
tentans and 28-d Hyalella azteca studies. All data has passed QA/QC review and are
considered final.
Please contact us if you have any questions regarding the data or methods utilized in
conducting the studies.
Daniel J. Farrar, M.S. Todd S. Bridges, Ph.D.
Research Biologist Research Biologist
Study Summary
Methods
The C. tentans 10-d and H. azteca 28-d studies were conducted following the methods
described in "Methods for Measuring the Toxicity and Bioaccumulation of Sediment-
associated Contaminants with Freshwater Invertebrates" (EPA/600/R-99/064, 2000).
Deviations from test method. Method deviations for the C. tentans study include a reduction
of the number of replicates per treatment from 8 to 7 and the use of dry weights rather than
ash-free dry weight measurements for biomass. The reduction in the number of replicates
was necessary in order to facilitate automated water change on the large number of sediment
samples tested. The test method suggests using ash-free fry dry weight to avoid the influence
of the grain size of ingested sediment on mass measurements. This is important when the
reference and test sediment have different grain size profiles. Since the sediment samples in
this study were compared to a reference with similar grain size, the influence of grain size on
mass measurements is minimal and the use of simple dry weight measurements is adequate.
Initial water quality for both tests was measured on 1 replicate per sediment sample at test
initiation rather than on all replicates as suggested in the test method. Experience has
demonstrated that when test setup occurs on day -1, water quality measurements on
individual replicates of a given sediment treatment on day 0 are essentially identical therefore
measurement on all replicates is not necessary. The method suggests measuring dissolved
oxygen daily for both tests since the method doesn't require aeration unless dissolved oxygen
measurements drops below 2.5 mg/L. Since the tests were aerated from test initiation to
avoid issues related to depleted oxygen, dissolved oxygen was only measured during the
course of the test if aeration was interrupted.
Statistical analyses. Statistical analyses were conducted for both studies following the
methods described in "Methods for Measuring the Toxicity and Bioaccumulation of
Sediment-associated Contaminants with Freshwater Invertebrates" and in "Evaluation of
Dredged material proposed for Discharge in the Waters of the U. S. -Testing Manual"
(EPA/823/B-98/004, 1998).
C. tentans survival. For each reference, all treatments that were not greater than 10%
different from the reference were removed from the data set prior to analysis. In addition, the
performance control and additional reference data were also removed. Analyses were
conducted using ANOVA on Ranks (Kruskal-Wallis) followed by Dunnett's mean
comparison to the reference.
C. tentans mass. The performance control and additional reference were removed from the
data set prior to analysis. Analyses were conducted using ANOVA on Ranks (Kruskal-
Wallis) for the BT-21 reference data set and one-way ANOVA for the BT-18 reference data
set.
H. azteca survival. For each reference, all treatments that were not greater than 10% different
from the reference were removed from the data set prior to analysis. In addition, the
performance control and additional reference data were also removed. Analyses were
conducted using one-way ANOVA followed by Dunnett's mean comparison to the reference.
2
H. azteca mass. The performance control and additional reference were removed from the
data set prior to analysis. Analyses were conducted using one-way ANOVA followed by
Dunnett's mean comparison to the reference.
Results
C. tentans 10-d experiment. Performance metrics were met with mean control survival
exceeding 92% (table 5). Water quality parameters fell within required ranges (tables 1-3).
Survival for the DNAPL and PITCH spiked sediments were significantly less than survival
observed in the BL18 and BL21 references (table 5). Survival in sediment BL11 was
significantly lower than survival observed in reference BL21. No effects on biomass were
observed for any of the sediments tested (table 6). Individual replicate survival and biomass
data are provided in table 4.
Kazteca 28-d experiment. Performance metrics were met with mean control survival
equaling 90% (table 12). Water quality parameters fell within required ranges (tables 7-10).
Survival for the PITCH spiked sediment was significantly less than survival observed in the
BL18 and BL21 references (table 12). Sediment BL9 biomass was significantly lower than
biomass observed for reference BL21 (table 13). Individual replicate survival and biomass
data are provided in table 11.
3
Table 1. C. tentans day 0 water quality

Sediment
Replicate
H
D.O. m /L 'Hardness
(PPM Ca CO3) 'Alkalinity
(PPM Ca CO3) 'Conductivit
y (µS) 'Ammonia
(mg/L)
Control A 8.28 8.59 90 120 340 1
DNAPL A 8.30 8.55 80 90 320 1
PITCH A 8.22 8.50 90 120 300 1
BL1 A 7.70 8.60 75 90 290 1
BL5 A 7.95 8.15 60 80 260 1
BL6 A 8.02 8.30 80 95 280 1
BL8 A 8.03 8.54 65 90 270 1
BL9 A 8.15 8.55 80 80 300 1
BL 10 A 7.90 8.47 90 80 260 1
BL 11 A 8.01 8.61 65 70 270 1
BL 12 A 8.10 8.40 65 90 260 1
BL14 A 8.12 8.59 70 95 330 1
BL18 A 8.26 8.51 100 100 360 1
BL20 A 8.30 8.51 80 100 330 1
BL21 A 7.97 8.42 65 80 280 1
Min 7.70 8.15 60 70 260 1
Max 8.30 8.61 100 120 360 1
Table 2. C. tentans day 10 water quality

Sediment
Replicate
H
D.O. m /L 'Hardness
PPM Ca CO3 'Alkalinity
PPM Ca C03 'Conductivity
S 'Ammonia
m /L
Control A 8.03 7.85 110 100 370 1
Control B 8.03 7.65
Control C 8.15 7.77
Control D 8.08 7.27
Control E 8.09 7.02
Control F 8.12 7.62
Control G 8.11 7.64
DNAPL A 8.23 8.19 110 130 340 2
DNAPL B 8.14 7.79
DNAPL C 8.30 8.05
DNAPL D 8.26 7.96
DNAPL E 8.29 7.83
DNAPL F 8.06 5.97
DNAPL G 8.22 8.01
PITCH A 8.04 7.10 80 110 333 2
PITCH B 8.17 8.21
PITCH C 8.21 8.82
PITCH D 8.25 8.23
PITCH E 8.22 7.80
PITCH F 8.22 8.16
PITCH G 8.31 8.00
'Replicate composite measurement
Table 2 (continued). C. tentans day 10 water quality

Sediment
Replica e
pH D.O.
m /L 'Hardness
PPM Ca CO3 'Alkalinity
PPM Ca C03 'Conductivity
S 'Ammonia
m /L
BL1 A 8.07 7.37 80 110 290 1
BL I B 7.93 7.18
BL I C 7.95 7.66
BL I D 7.98 7.87
BL1 E 7.93 7.58
BL I F 7.75 5.00
BL I G 7.73 6.87
BL5 A 7.80 7.18 75 100 270 1
BL5 B 7.88 7.89
BL5 C 7.87 7.83
BL5 D 7.88 7.67
BL5 E 7.70 4.77
BL5 F 7.77 7.70
BL5 G 7.81 7.75
BL6 A 7.88 7.85 75 90 290 1
BL6 B 7.92 7.99
BL6 C 7.93 7.58
BL6 D 7.94 7.58
BL6 E 7.97 7.92
BL6 F 7.99 7.56
BL6 G 8.01 7.87
'Replicate composite measurement
Table 2 (continued). C. tentans day 10 water quality

Sediment
Replica e
pH D.O.
m /L 'Hardness
PPM Ca C03 'Alkalinity
PPM Ca C03 'Conductivity
S 'Ammonia
m /L
BL8 A 7.99 7.91 65 90 280 1
BL8 B 7.92 7.44
BL8 C 7.91 7.79
BL8 D 7.92 7.84
BL8 E 7.88 7.46
BL8 F 7.88 7.61
BL8 G 7.82 7.01
BL9 A 7.88 7.87 85 90 310 1
BL9 B 7.90 7.18
BL9 C 7.87 7.38
BL9 D 7.68 5.00
BL9 E 7.84 7.83
BL9 F 7.98 7.70
BL9 G 7.98 7.72
BL10 A 7.97 7.84 60 80 270 1
BL10 B 7.94 7.71
BL 10 C 7.82 6.03
BL10 D 7.88 7.96
BL10 E 7.89 7.68
BL10 F 7.86 7.46
BL 10 G 7.87 7.66
'Replicate composite measurement
Table 2 (continued). C. tentans day 10 water quality

Sediment
Replica e
pH
D.O. m /L 'Hardness
PPM Ca CO3 'Alkalinity
PPM Ca C03 'Conductivity
S 'Ammonia
m /L
BL11 A 7.85 7.58 75 80 260 1
BL11 B 7.86 7.73
BL 11 C 7.90 7.91
BL11 D 7.88 7.61
BL11 E 7.92 7.91
BL11 F 7.81 6.60
BL 11 G 7.83 7.70
BL12 A 7.78 5.28 65 70 270 1
BL12 B 7.80 7.39
BL12 C 7.85 7.75
BL12 D 7.81 7.06
BL 12 E 7.82 7.29
BL12 F 7.87 7.35
BL12 G 7.86 7.51
BL14 A 7.81 6.88 70 90 290 1
BL14 B 7.78 7.06
BL 14 C 7.90 7.79
BL14 D 7.93 7.51
BL 14 E 7.92 7.64
BL14 F 7.80 6.40
BL14 G 7.84 7.50
'Replicate composite measurement
Table 2 (continued). C. tentans day 10 water quality

Sediment
Replicate
H
D.O. m /L 'Hardness
PPM Ca CO3 'Alkalinity
PPM Ca C03 'Conductivity
S 'Ammonia
m /L
BL18 A 7.91 7.76 85 100 310 1
BL 18 B 7.93 7.60
BL 18 C 7.95 7.46
BL18 D 7.85 4.07
BL 18 E 7.94 7.77
BL 18 F 8.05 7.75
BL 18 G 8.07 7.43
BL20 A 8.08 7.98 80 70 300 1
BL20 B 8.10 8.11
BL20 C 8.12 7.99
BL20 D 8.14 7.82
BL20 E 8.12 7.84
BL20 F 8.10 7.50
BL20 G 8.12 7.85
BL21 A 8.12 7.85 65 90 260 1
BL21 B 8.01 4.97
BL21 C 7.96 7.80
BL21 D 7.92 7.70
BL21 E 7.92 7.37
BL21 F 7.85 7.14
BL21 G 7.83 7.73
Min 7.68 4.07 60 70 260 1
Max 8.31 8.82 110 130 370 2
'Replicate composite measurement
Table 3. C. tentans Daily Temperatures
Da Statistics
Sediment 0 1 2 3 4 5 6 7 8 9 10 Mean Std. Dev. Min Max
Control 21.5 21.8 21.9 21.8 21.9 22.5 22.6 20.6 22.1 22.3 23.4 22.0 0.7 20.6 23.4
DNAPL 21.8 22.0 22.0 21.9 22.0 22.4 22.6 20.6 23.1 22.3 23.2 22.2 0.7 20.6 23.2
PITCH 21.8 21.9 21.9 22.0 22.0 22.4 22.5 20.5 23.2 22.3 23.2 22.2 0.7 20.5 23.2
BL I 21.5 21.9 22.0 21.8 21.9 22.4 22.5 20.9 23.1 22.3 23.4 22.2 0.7 20.9 23.4
BL5 21.7 22.0 21.9 21.9 22.0 22.3 22.6 21.3 23.1 22.3 23.4 22.2 0.6 21.3 23.4
BL6 21.5 21.6 21.8 21.8 22.0 22.4 22.5 20.6 23.2 22.3 23.5 22.1 0.8 20.6 23.5
BL8 21.6 22.0 22.0 22.0 21.9 22.4 22.5 21.0 23.2 22.3 23.4 22.2 0.7 21.0 23.4
BL9 21.7 21.8 21.8 21.9 22.0 22.4 22.5 21.1 23.1 22.3 23.5 22.2 0.7 21.1 23.5
BL10 21.9 22.0 22.0 21.9 21.9 22.4 22.5 21.1 23.2 22.3 23.4 22.2 0.6 21.1 23.4
BL11 21.2 22.0 22.0 21.9 22.0 22.3 22.5 21.2 23.1 22.3 23.2 22.2 0.6 21.2 23.2
BL12 21.8 21.8 22.0 21.8 21.9 22.4 22.5 21.2 23.2 22.3 23.2 22.2 0.6 21.2 23.2
BL14 21.7 21.9 21.9 21.9 21.9 22.4 22.5 21.4 23.1 22.3 23.4 22.2 0.6 21.4 23.4
BL18 21.8 21.9 21.8 21.9 22.0 22.3 22.4 21.2 23.2 22.3 23.2 22.2 0.6 21.2 23.2
BL20 21.8 21.9 21.8 21.9 22.0 22.4 22.5 20.5 23.1 22.3 23.2 22.1 0.7 20.5 23.2
BL21 21.8 22.0 21.8 21.8 21.9 22.4 22.5 20.5 23.2 22.3 23.2 22.1 0.7 20.5 23.2
10
Table 4. C. tentans 10-d endpoint data

Treatment

Replicate

Survival

% Survival

# on pan
pan weight
(g) pan &
animal
weight (g) individual
dry weight
(mg)
Control A 9 90.0% 9 0.0950 0.1157 2.30
Control B 10 100.0% 10 0.0942 0.1142 2.00
Control c 10 100.0% 10 0.0967 0.1153 1.86
Control D 9 90.0% 9 0.0855 0.1107 2.80
Control E 10 100.0% 10 0.1023 0.1263 2.40
Control F 10 100.0% 10 0.0905 0.1115 2.10
Control G 7 70.0% 7 0.0897 0.1079 2.60
DNAPL A 5 50.0% 5 0.0977 0.1076 1.98
DNAPL B 5 50.0% 5 0.0978 0.1073 1.90
DNAPL C 6 60.0% 6 0.0965 0.1147 3.03
DNAPL D 8 80.0% 8 0.0924 0.1214 3.63
DNAPL E 6 60.0% 6 0.1013 0.1143 2.17
DNAPL F 6 60.0% 6 0.0913 0.1086 2.88
DNAPL G 7 70.0% 7 0.0923 0.1097 2.49
PITCH A 6 60.0% 6 0.0888 0.0973 1.42
PITCH B 6 60.0% 6 0.0883 0.1019 2.27
PITCH C 6 60.0% 6 0.1024 0.1130 1.77
PITCH D 7 70.0% 7 0.0941 0.1050 1.56
PITCH E 5 50.0% 5 0.0992 0.1084 1.84
PITCH F 7 70.0% 7 0.0946 0.1071 1.79
PITCH G 5 50.0% 5 0.0917 0.1011 1.88
11
Table 4 (continued). C. tentans 10-d endpoint data

Treatment

Replicate

Survival

% Survival

# on an
pan weight pan &
animal
wei ht individual
dry weight
m
BL-1 A 8 80.0% 8 0.0919 0.1083 2.05
BL-1 B 9 90.0% 9 0.0982 0.1202 2.44
BL-1 C 10 100.0% 10 0.1010 0.1234 2.24
BL-1 D 10 100.0% 10 0.0857 0.1050 1.93
BL-1 E 10 100.0% 10 0.0937 0.1163 2.26
BL-1 F 10 100.0% 10 0.0909 0.1096 1.87
BL-1 G 9 90.0% 9 0.1036 0.1163 1.41
BL5 A 9 90.0% 9 0.0960 0.1159 2.21
BL5 B 10 100.0% 10 0.0838 0.1056 2.18
BL5 C 8 80.0% 8 0.0754 0.0936 2.28
BL5 D 10 100.0% 10 0.0954 0.1167 2.13
BL5 E 10 100.0% 10 0.0847 0.0976 1.29
BL5 F 10 100.0% 10 0.0905 0.1205 3.00
BL5 G 10 100.0% 8 0.1005 0.1283 3.48
BL6 A 5 50.0% 5 0.0915 0.1068 3.06
BL6 B 8 80.0% 8 0.1871 0.2032 2.01
BL6 C 10 100.0% 10 0.1917 0.2121 2.04
BL6 D 10 100.0% 10 0.0998 0.1230 2.32
BL6 E 7 70.0% 7 0.0874 0.0996 1.74
BL6 F 10 100.0% 9 0.0949 0.1252 3.37
BL6 G 9 90.0% 6 0.0800 0.0950 2.50
12
Table 4 (continued). C. tentans 10-d endpoint data

Treatment

Replicate

Survival

% Survival

# on an
pan weight pan &
animal
wei ht individual
dry weight
m
BL8 A 10 100.0% 10 0.0972 0.1163 1.91
BL8 B 8 80.0% 8 0.0907 0.1109 2.53
BL8 C 8 80.0% 8 0.0933 0.1052 1.49
BL8 D 10 100.0% 10 0.0891 0.1133 2.42
BL8 E 10 100.0% 10 0.0885 0.1136 2.51
BL8 F 10 100.0% 10 0.0986 0.1215 2.29
BL8 G 10 100.0% 10 0.0895 0.1127 2.32
BL9 A 9 90.0% 9 0.0879 0.1073 2.16
BL9 B 8 80.0% 8 0.0944 0.1065 1.51
BL9 C 9 90.0% 8 0.0914 0.1143 2.86
BL9 D 10 100.0% 10 0.0948 0.1181 2.33
BL9 E 6 60.0% 6 0.0933 0.1081 2.47
BL9 F 6 60.0% 6 0.0951 0.1102 2.52
BL9 G 10 100.0% 10 0.1020 0.1223 2.03
BL10 A 10 100.0% 10 0.1050 0.1296 2.46
BL 10 B 9 90.0% 9 0.0890 0.1091 2.23
BL 10 C 10 100.0% 10 0.0998 0.1219 2.21
BL10 D 6 60.0% 6 0.0803 0.0932 2.15
BL10 E 5 50.0% 5 0.0948 0.1113 3.30
BL 10 F 4 40.0% 4 0.0913 0.1084 4.28
BL10 G 10 100.0% 10 0.0832 0.1092 2.60
13
Table 4 (continued). C. tentans 10-d endpoint data

Treatment

Replicate

Survival

% Survival

# on an
pan weight pan &
animal
wei ht individual
dry weight
m
BL 11 A 10 100.0% 10 0.0911 0.1135 2.24
BL11 B 8 80.0% 8 0.0938 0.1125 2.34
BL 11 C 6 60.0% 6 0.0986 0.1167 3.02
BL11 D 1 10.0% 1 0.0955 0.0980 2.50
BL11 E 5 50.0% 5 0.0866 0.1065 3.98
BL 11 F 7 70.0% 7 0.0906 0.1092 2.66
BL11 G 7 70.0% 7 0.0956 0.1139 2.61
BL12 A 8 80.0% 8 0.0924 0.1085 2.01
BL 12 B 7 70.0% 7 0.0908 0.1112 2.91
BL12 C 9 90.0% 9 0.0903 0.1137 2.60
BL 12 D 9 90.0% 9 0.0998 0.1224 2.51
BL 12 E 10 100.0% 10 0.0872 0.1182 3.10
BL 12 F 9 90.0% 9 0.0982 0.1128 1.62
BL 12 G 9 90.0% 9 0.0973 0.1220 2.74
BL 14 A 7 70.0% 7 0.0953 0.1124 2.44
BL 14 B 4 40.0% 4 0.0995 0.1092 2.43
BL14 C 10 100.0% 10 0.0994 0.1179 1.85
BL14 D 9 90.0% 9 0.0979 0.1250 3.01
BL14 E 9 90.0% 9 0.0936 0.1123 2.08
BL14 F 10 100.0% 10 0.0955 0.1282 3.27
BL14 G 10 100.0% 10 0.0863 0.1122 2.59
14
Table 4 (continued). C. tentans 10-d endpoint data

Treatment

Replicate

Survival

% Survival

# on an
pan weight pan &
animal
wei ht individual
dry weight
m
BL18 A 8 80.0% 8 0.0924 0.1131 2.59
BL18 B 10 100.0% 10 0.0940 0.1133 1.93
BL 18 C 8 80.0% 8 0.0931 0.1153 2.78
BL 18 D 8 80.0% 8 0.0962 0.1142 2.25
BL18 E 10 100.0% 10 0.0939 0.1117 1.78
BL18 F 10 100.0% 10 0.0929 0.1114 1.85
BL 18 G 8 80.0% 8 0.0897 0.1114 2.71
BL20 A 10 100.0% 10 0.0890 0.1036 1.46
BL20 B 10 100.0% 10 0.0851 0.0997 1.46
BL20 C 9 90.0% 9 0.0892 0.1069 1.97
BL20 D 7 70.0% 7 0.0890 0.1085 2.79
BL20 E 10 100.0% 10 0.0910 0.1094 1.84
BL20 F 5 50.0% 5 0.0870 0.1037 3.34
BL20 G 10 100.0% 10 0.0896 0.1066 1.70
BL21 A 10 100.0% 10 0.0901 0.1081 1.80
BL21 B 10 100.0% 10 0.0959 0.1119 1.60
BL21 C 10 100.0% 10 0.0951 0.1169 2.18
BL21 D 10 100.0% 10 0.0802 0.1004 2.02
BL21 E 10 100.0% 10 0.0894 0.1168 2.74
BL21 F 10 100.0% 10 0.0858 0.1079 2.21
BL21 G 10 100.0% 10 0.0971 0.1182 2.11
15
Table 5. C. tentans 10-d survival
Treatment Mean % S.D. C.V. n
Control 92.9% 11.1% 11.98% 7
DNAPL A, B 61.4% 10.7% 17.40% 7
PITCH A, B 60.0% 8.2% 13.61% 7
BL1 94.3% 7.9% 8.34% 7
BL5 95.7% 7.9% 8.22% 7
BL6 84.3% 19.0% 22.57% 7
BL8 94.3% 9.8% 10.35% 7
BL9 82.9% 17.0% 20.57% 7
BL10 77.1% 26.3% 34.06% 7
BL11 B 62.9% 28.1% 44.73% 7
BL12 87.1% 9.5% 10.92% 7
BL14 84.3% 22.3% 26.40% 7
BL18 88.6% 10.7% 12.07% 7
BL20 87.1% 19.8% 22.68% 7
BL21 100.0% 0.0% 0.00% 7
A= Significantly different from BL 18 reference
B=Significantly different from BL21 reference
16
Table 6. C. tentans 10-d mass
Treatment Mean m S.D. C.V. n
Control 2.3 0.3 14.60% 7
DNAPL 2.6 0.6 24.45% 7
PITCH 1.8 0.3 15.01% 7
BL I 2.0 0.3 16.68% 7
BL5 2.4 0.7 29.45% 7
BL6 2.4 0.6 24.24% 7
BL8 2.2 0.4 17.18% 7
BL9 2.3 0.4 18.87% 7
BL10 2.7 0.8 28.41% 7
BL11 2.8 0.6 21.42% 7
BL12 2.5 0.5 20.71% 7
BL14 2.5 0.5 19.60% 7
BL18 2.3 0.4 18.73% 7
BL20 2.1 0.7 34.40% 7
BL21 2.1 0.4 17.15% 7
A= Significantly different from BL 18 reference
B=Significantly different from BL21 reference
17
Table 7. H. azteca Day 0 water quality

Sediment
Replicate
H
D.O. m /L 'Hardness
PPM Ca CO3 'Alkalinity
PPM Ca CO3 'Conductivity
S 'Ammonia
m /L
Control A 8.18 7.72 90 110 280 1
DNAPL A 8.20 7.84 80 110 270 2
PITCH A 8.14 7.85 60 100 220 1
BL I A 8.06 7.53 100 75 240 1
BL5 A 7.98 7.60 100 80 200 1
BL6 A 7.99 7.86 100 90 190 1
BL8 A 7.92 7.53 60 70 200 1
BL9 A 8.05 7.86 65 70 220 1
BL10 A 7.92 8.02 80 60 200 1
BL11 A 8.02 7.96 80 60 200 1
BL12 A 8.08 7.92 120 60 210 1
BL14 A 8.03 7.94 90 80 220 1
BL 18 A 7.60 7.72 120 120 270 1
BL20 A 8.10 7.91 120 85 250 1
BL21 A 7.90 7.74 110 60 200 1
Min 7.60 7.53 60 60 190 1
Max 8.20 8.02 120 120 280 2
18
Table 8. H. azteca day 28 water quality

Sediment
Replicate
H
D.O. m /L 'Hardness
PPM Ca C03 'Alkalinity
PPM Ca C03 'Conductivity
S 'Ammonia
m /L
Control A 8.26 7.86 100 80 260 1
Control B 8.23 8.30
Control C 8.28 8.01
Control D 8.30 8.06
Control E 8.39 8.51
Control F 8.25 8.02
DNAPL A 8.26 8.21 70 80 260 1
DNAPL B 8.26 7.89
DNAPL C 8.35 8.35
DNAPL D 8.14 7.47
DNAPL E 8.30 8.22
DNAPL F 8.35 8.11
PITCH A 8.39 8.69 80 80 260 1
PITCH B 8.29 8.14
PITCH C 8.28 8.03
PITCH D 8.29 7.99
PITCH E 8.28 8.11
PITCH F 8.28 8.09
'Replicate composite measurement
19
Table 8 (continued). H. azteca Day 28 Water Quality

Sediment
Replicate
H
D.O. m /L 'Hardness
PPM Ca C03 'Alkalinity
PPM Ca C03 'Conductivity
S 'Ammonia
m /L
BL1 A 8.16 8.29 80 70 260 1
BL1 B 8.30 8.54
BL1 C 8.32 8.50
BL1 D 8.23 8.08
BL1 E 8.26 8.56
BL1 F 8.20 8.17
BL5 A 8.30 8.36 70 80 250 1
BL5 B 8.25 8.43
BL5 C 8.29 8.07
BL5 D 8.29 8.28
BL5 E 8.23 7.81
BL5 F 8.29 8.21
BL6 A 8.25 8.25 70 80 260 1
BL6 B 8.29 8.24
BL6 C 8.26 7.98
BL6 D 8.34 8.20
BL6 E 8.28 8.12
BL6 F 8.36 8.03
'Replicate composite measurement
20
Table 8 (continued). H. azteca day 28 water quality

Sediment
Replicate
H
D.O. m /L 'Hardness
PPM Ca CO3 'Alkalinity
PPM Ca C03 'Conductivity
S 'Ammonia
m /L
BL8 A 8.29 8.47 70 70 250 1
BL8 B 8.27 7.99
BL8 C 8.21 8.50
BL8 D 8.33 8.05
BL8 E 8.25 8.17
BL8 F 8.26 8.24
BL9 A 8.05 8.28 70 80 260 1
BL9 B 8.30 8.63
BL9 C 8.31 8.53
BL9 D 8.32 8.09
BL9 E 8.26 8.02
BL9 F 8.32 8.18
BL 10 A 8.35 8.74 70 70 240 1
BL 10 B 8.25 8.77
BL 10 C 8.26 8.63
BL 10 D 8.23 8.07
BL 10 E 8.33 8.17
BL10 F 8.30 8.70
'Replicate composite measurement
21
Table 8 (continued). H. azteca day 28 water quality

Sediment
Replicate
H
D.O. m /L 'Hardness
PPM Ca CO3 'Alkalinity
PPM Ca C03 'Conductivity
S 'Ammonia
m /L
BL 11 A 8.27 8.00 70 70 270 1
BL 11 B 8.26 8.10
BL 11 C 8.25 8.61
BL 11 D 8.27 8.35
BL 11 E 8.22 7.98
BL 11 F 8.36 8.43
BL12 A 8.25 7.82 70 80 250 1
BL12 B 8.26 8.30
BL 12 C 8.34 8.47
BL 12 D 8.11 7.99
BL 12 E 8.29 8.57
BL12 F 8.39 8.14
BL 14 A 8.25 8.43 70 90 290 1
BL 14 B 8.26 8.43
BL 14 C 8.16 8.37
BL14 D 8.35 8.28
BL 14 E 8.26 8.15
BL14 F 8.36 8.52
'Replicate composite measurement
22
Table 8 (continued). H. azteca day 28 water quality

Sediment
Replicate
H
D.O. m /L 'Hardness
PPM Ca CO3 'Alkalinity
PPM Ca C03 'Conductivity
S 'Ammonia
m /L
BL18 A 8.31 8.61 70 80 260 1
BL18 B 8.08 8.14
BL18 C 8.35 8.43
BL18 D 8.33 8.27
BL18 E 8.33 8.00
BL18 F 8.23 8.40
BL20 A 8.36 8.68 70 90 260 1
BL20 B 8.26 8.55
BL20 C 8.27 8.51
BL20 D 8.30 8.77
BL20 E 8.05 8.30
BL20 F 8.39 8.37
BL21 A 8.19 8.50 60 80 250 1
BL21 B 8.20 8.51
BL21 C 8.21 8.23
BL21 D 8.03 7.88
BL21 E 8.28 8.59
BL21 F 8.25 7.99
Min 8.03 7.47 60 70 240 1
Max 8.39 8.77 100 90 290 1
'Replicate composite measurement
23
Table 9. H. azteca daily replicate temperatures
DAY
Sediment 0 1 2 3 4 5 6 7 8 9 10 11 12 13 14
Control 22.0 21.9 23.2 23.7 23.1 23.2 23.1 23.4 23.2 23.3 23.4 23.3 23.3 23.1 23.2
DNAPL 22.0 21.9 23.2 23.7 23.1 23.2 23.1 23.4 23.2 23.3 23.4 23.3 23.3 23.2 23.2
PITCH 22.0 21.9 23.2 23.7 23.1 23.2 23.1 23.4 23.2 23.2 23.4 23.3 23.3 23.1 23.2
BL1 22.1 21.9 23.1 23.2 23.1 23.2 23.1 23.4 23.2 23.3 23.4 23.2 23.3 23.1 23.2
BL5 22.0 21.8 23.2 23.2 23.5 23.2 23.1 23.4 23.2 23.2 23.3 23.2 23.2 23.1 23.2
BL6 22.0 21.9 23.1 23.7 23.5 23.1 23.2 23.4 23.1 23.2 23.4 23.2 23.2 23.2 23.2
BL8 21.9 22.0 23.2 23.2 23.5 23.1 23.1 23.4 23.2 23.3 23.4 23.3 23.2 23.2 23.1
BL9 22.0 21.9 23.1 23.7 23.1 23.2 23.1 23.4 23.2 23.2 23.4 23.2 23.3 23.1 23.2
BL10 22.0 21.9 23.2 23.7 23.5 23.1 23.1 23.4 23.2 23.3 23.4 23.3 23.3 23.1 23.2
BL11 21.9 21.9 23.2 23.7 23.5 23.1 23.1 23.4 23.2 23.3 23.3 23.3 23.3 23.1 23.1
BL12 21.9 21.9 23.2 23.7 23.4 23.0 23.1 23.4 23.2 23.3 23.3 23.3 23.3 23.1 23.1
BL14 22.0 21.9 23.2 23.7 23.5 23.0 23.1 23.4 23.2 23.2 23.4 23.2 23.3 23.1 23.1
BL18 22.0 21.9 23.2 23.7 23.5 23.1 23.2 23.4 23.1 23.3 23.3 23.3 23.3 23.2 23.1
BL20 22.0 21.8 23.2 23.7 23.5 23.0 23.1 23.4 23.1 23.3 23.3 23.3 23.3 23.1 23.1
BL21 22.0 21.8 23.1 23.7 23.5 23.0 23.1 23.4 23.1 23.3 23.3 23.3 23.3 23.1 23.1
24
Table 9 (continued). H. azteca daily replicate temperatures
DAY Statistics
15 16 17 18 19 20 21 22 23 24 25 26 27 28 Mean Std. Dev. Min Max
23.3 23.5 23.5 23.4 23.2 23.4 23.5 23.5 23.4 23.5 23.4 23.5 23.5 23.4 23.3 0.4 21.9 23.7
23.3 23.5 23.5 23.6 23.3 23.4 23.4 23.5 23.4 23.5 23.4 23.4 23.2 23.4 23.3 0.4 21.9 23.7
23.3 23.5 23.5 23.4 23.4 23.4 23.4 23.4 23.5 23.5 23.4 23.4 23.2 23.4 23.2 0.4 21.9 23.7
23.3 23.4 23.5 23.3 23.3 23.4 23.4 23.4 23.5 23.5 23.3 23.4 23.4 23.4 23.2 0.4 21.9 23.5
23.3 23.5 23.5 23.4 23.4 23.3 23.4 23.5 23.5 23.5 23.3 23.4 23.3 23.4 23.2 0.4 21.8 23.5
23.3 23.5 23.5 23.3 23.4 23.3 23.3 23.5 23.5 23.5 23.3 23.4 23.2 23.4 23.2 0.4 21.9 23.7
23.3 23.5 23.5 23.3 23.4 23.4 23.3 23.4 23.5 23.5 23.3 23.4 23.4 23.4 23.2 0.4 21.9 23.5
23.3 23.4 23.5 23.4 23.4 23.4 23.3 23.5 23.4 23.5 23.3 23.4 23.3 23.4 23.2 0.4 21.9 23.7
23.3 23.5 23.5 23.4 23.4 23.4 23.3 23.4 23.4 23.5 23.3 23.4 23.2 23.4 23.2 0.4 21.9 23.7
23.3 23.5 23.5 23.3 23.4 23.3 23.4 23.5 23.5 23.5 23.3 23.4 23.3 23.4 23.2 0.4 21.9 23.7
23.3 23.5 23.5 23.3 23.4 23.3 23.3 23.5 23.5 23.5 23.3 23.4 23.3 23.4 23.2 0.4 21.9 23.7
23.4 23.4 23.4 23.6 23.4 23.3 23.3 23.5 23.5 23.6 23.3 23.4 23.5 23.5 23.3 0.4 21.9 23.7
23.4 23.4 23.4 23.2 23.4 23.4 23.3 23.5 23.5 23.6 23.3 23.4 23.4 23.5 23.3 0.4 21.9 23.7
23.3 23.4 23.5 23.4 23.4 23.3 23.3 23.5 23.4 23.6 23.4 23.5 23.4 23.5 23.2 0.4 21.8 23.7
23.3 23.5 23.5 23.2 23.4 23.3 23.3 23.4 23.5 23.6 23.4 23.5 23.4 23.4 23.2 0.4 21.8 23.7
25
Table 10. Weekly conductivity ( µS) measurements
WEEK
Sediment 1 2 3
Control 280 200 240
DNAPL 270 240 260
PITCH 270 220 230
BL1 280 230 230
BL5 270 240 220
BL6 220 260 280
BL8 240 230 260
BL9 230 200 270
BL 10 260 270 260
BL 11 260 210 150
BL 12 250 220 240
BL 14 260 220 260
BL18 260 230 270
BL20 280 240 240
BL21 260 210 230
Min 220 200 150
Max 280 270 280
26
Table 11. H. azteca 28-d endpoint data

Treatment

Replicate

Survival

% Survival

# on pan
pan weight
(g) pan &
animal
weight (g) individual
dry weight
(mg)
Control A 10 100% 10 0.0734 0.0756 0.22
Control B 9 90% 9 0.0726 0.0745 0.21
Control c 8 80% 8 0.0754 0.0774 0.25
Control D 10 100% 10 0.0860 0.0876 0.16
Control E 10 100% 10 0.0812 0.0828 0.16
Control F 10 100% 10 0.0966 0.0982 0.16
DNAPL A 10 100% 9 0.0852 0.0878 0.29
DNAPL B 9 90% 9 0.0894 0.0916 0.24
DNAPL C 7 70% 7 0.0826 0.0849 0.33
DNAPL D 10 100% 10 0.0920 0.0933 0.13
DNAPL E 10 100% 10 0.0685 0.0703 0.18
DNAPL F 10 100% 9 0.0902 0.0936 0.38
Pitch A 0 0% 0
Pitch B 0 0% 0
Pitch c 4 40% 4 0.0878 0.0889 0.28
Pitch D 0 0% 0
Pitch E 0 0% 0
Pitch F 0 0% 0
27
Table II (continued). H. azteca 28-d endpoint data

Treatment

Replicate

Survival

% Survival

# on an
pan weight pan &
animal
wei ht individual
dry weight
m
BL-1 A 8 80% 8 0.0817 0.0836 0.24
BL-1 B 10 100% 9 0.0763 0.0785 0.24
BL-1 C 10 100% 9 0.0795 0.0818 0.26
BL-1 D 10 100% 10 0.0661 0.0677 0.16
BL-1 E 10 100% 9 0.0811 0.0828 0.19
BL-1 F 10 100% 10 0.0751 0.0768 0.17
BL5 A 9 90% 8 0.0798 0.0818 0.25
BL5 B 10 100% 10 0.0807 0.0824 0.17
BL5 C 7 70% 7 0.0754 0.0770 0.23
BL5 D 7 70% 7 0.0830 0.0841 0.16
BL5 E 8 80% 7 0.0656 0.0673 0.24
BL5 F 8 80% 8 0.0841 0.0857 0.20
BL6 A 6 60% 6 0.0754 0.0773 0.32
BL6 B 8 80% 8 0.0880 0.0893 0.16
BL6 C 7 70% 7 0.1037 0.1054 0.24
BL6 D 9 90% 9 0.0690 0.0706 0.18
BL6 E 10 100% 8 0.0681 0.0697 0.20
BL6 F 5 50% 5 0.0725 0.0733 0.16
28
Table II (continued). H. azteca 28-d endpoint data

Treatment

Replicate

Survival

% Survival

# on an
pan weight pan &
animal
wei ht individual
dry weight
m
BL8 A 10 100% 10 0.0718 0.0737 0.19
BL8 B 6 60% 6 0.0777 0.0798 0.35
BL8 C 6 60% 6 0.0903 0.0915 0.20
BL8 D 8 80% 8 0.0712 0.0723 0.14
BL8 E 7 70% 7 0.0890 0.0905 0.21
BL8 F 9 90% 9 0.0876 0.0888 0.13
BL9 A 10 100% 10 0.0896 0.0911 0.15
BL9 B 9 90% 9 0.0894 0.0906 0.13
BL9 C 9 90% 9 0.0792 0.0802 0.11
BL9 D 8 80% 8 0.0827 0.0837 0.13
BL9 E 10 100% 9 0.0915 0.0927 0.13
BL9 F 10 100% 10 0.0848 0.0859 0.11
BL10 A 9 90% 9 0.0850 0.0861 0.12
BL10 B 10 100% 10 0.0850 0.0860 0.10
BL 10 C 8 80% 8 0.1003 0.1023 0.25
BL 10 D 10 100% 10 0.1066 0.1078 0.12
BL10 E 7 70% 7 0.0940 0.0950 0.14
BL 10 F 10 100% 10 0.1107 0.1126 0.19
29
Table II (continued). H. azteca 28-d endpoint data

Treatment

Replicate

Survival

% Survival

# on an
pan weight pan &
animal
wei ht individual
dry weight
m
BL 11 A 10 100% 10 0.1190 0.1212 0.22
BL11 B 10 100% 10 0.0955 0.0974 0.19
BL 11 C 10 100% 10 0.1084 0.1108 0.24
BL11 D 9 90% 9 0.1039 0.1048 0.10
BL 11 E 9 90% 9 0.1044 0.1061 0.19
BL 11 F 10 100% 10 0.0954 0.0966 0.12
BL 12 A 10 100% 10 0.0907 0.0923 0.16
BL 12 B 9 90% 9 0.0924 0.0940 0.18
BL12 C 9 90% 9 0.1069 0.1101 0.36
BL 12 D 9 90% 9 0.1068 0.1081 0.14
BL 12 E 10 100% 9 0.1027 0.1042 0.17
BL 12 F 9 90% 9 0.0912 0.0926 0.16
BL 14 A 10 100% 10 0.0874 0.0897 0.23
BL14 B 6 60% 6 0.0828 0.0837 0.15
BL 14 C 10 100% 10 0.0966 0.0990 0.24
BL 14 D 10 100% 10 0.0824 0.0842 0.18
BL 14 E 6 60% 6 0.0780 0.0789 0.15
BL 14 F 10 100% 10 0.0948 0.0967 0.19
30
Table II (continued). H. azteca 28-d endpoint data

Treatment

Replicate

Survival

% Survival

# on an
pan weight pan &
animal
wei ht individual
dry weight
m
BL18 A 7 70% 7 0.0988 0.1002 0.20
BL 18 B 8 80% 8 0.1001 0.1020 0.24
BL18 C 10 100% 10 0.0930 0.0942 0.12
BL 18 D 9 90% 9 0.0777 0.0790 0.14
BL 18 E 9 90% 9 0.0793 0.0806 0.14
BL18 F 8 80% 8 0.0855 0.0867 0.15
BL20 A 9 90% 9 0.0794 0.0817 0.26
BL20 B 9 90% 9 0.0677 0.0698 0.23
BL20 C 10 100% 10 0.0882 0.0903 0.21
BL20 D 8 80% 8 0.0835 0.0851 0.20
BL20 E 9 90% 9 0.0698 0.0719 0.23
BL20 F 8 80% 8 0.0882 0.0905 0.29
BL21 A 10 100% 10 0.0767 0.0796 0.29
BL21 B 8 80% 8 0.0812 0.0829 0.21
BL21 C 10 100% 10 0.0790 0.0813 0.23
BL21 D 5 50% 5 0.0720 0.0737 0.34
BL21 E 10 100% 10 0.0933 0.0948 0.15
BL21 F 10 100% 9 0.1004 0.1022 0.20
31
Table 12. H. azteca 28-d survival
Treatment Mean % S.D. C.V. n
Control 95.0% 8.4% 8.81% 6
DNAPL 93.3% 12.1% 12.98% 6
PITCH A, B 6.7% 16.3% 244.95% 6
BL I 96.7% 8.2% 8.45% 6
BL5 81.7% 11.7% 14.31% 6
BL6 75.0% 18.7% 24.94% 6
BL8 76.7% 16.3% 21.30% 6
BL9 93.3% 8.2% 8.75% 6
BL10 90.0% 12.6% 14.05% 6
BL11 96.7% 5.2% 5.34% 6
BL12 93.3% 5.2% 5.53% 6
BL14 86.7% 20.7% 23.83% 6
BL18 85.0% 10.5% 12.34% 6
BL20 88.3% 7.5% 8.52% 6
BL21 88.3% 20.4% 23.11% 6
A=Significantly different from BL-18 reference
B=Significantly different from BL-21 reference
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Table 13. H. azteca 28-d mass
Treatment Mean m S.D. C.V. n
Control 0.19 0.04 20.11% 6
DNAPL 0.26 0.09 35.88% 6
PITCH # 0.26 N/A N/A 1
BL I 0.21 0.04 19.76% 6
BL5 0.21 0.04 18.61% 6
BL6 0.21 0.06 28.85% 6
BL8 0.20 0.08 38.59% 6
BL9 B 0.13 0.02 11.95% 6
BL10 0.15 0.06 36.36% 6
BL11 0.18 0.06 31.35% 6
BL12 0.19 0.08 41.51% 6
BL14 0.19 0.04 20.25% 6
BL18 0.17 0.04 26.34% 6
BL20 0.24 0.03 13.39% 6
BL21 0.24 0.07 28.61% 6
B=Significantly different from BL-21 Reference
#=Excluded from analysis due to significant mortality in the replicate
33