600R08045
f/EPA
United States
Environmental Protection
Agency
Toxicology
-------
EPA/600/R-08/045
September 2008
ENDANGERED AQUATIC VERTEBRATES: COMPARATIVE
AND PROBABILISTIC-BASED TOXICOLOGY
by
Foster L. Mayer1'5, Denny R. Buckler2, F. James Dwyer3, Mark R. Ellersieck4, Linda C.
Sappington2, John M. Besser2, and Christine M. Bridges2
1U.S. Environmental Protection Agency
Office of Research and Development
National Health and Environmental Effects Research Laboratory
Gulf Ecology Division
Gulf Breeze, FL 32561
2U.S. Geological Survey
Columbia Environmental Research Center
Columbia, MO 65201
3U.S. Fish and Wildlife Service
Columbia, MO 65201
4University of Missouri-Columbia
College of Agriculture, Food and Natural Resources
Agricultural Experiment Station-Statistics
Columbia, MO 65211
5Present address:
8069 Constitution Rd.
Las Cruces, NM 88007
U.S. Environmental Protection Agency
Office of Research and Development
1200 Pennsylvania Avenue, NW
Washington, DC 20460
-------
-------
TABLE OF CONTENTS
Notice iv
Abstract v
Figures ., vi
Tables vii
Acknowledgements viii
1. Introduction 1
2. Methods .- 2
Toxicity Tests 2
Statistical Analyses 2
Acute Toxicity 2
Chronic Toxicity 3
Estimating Toxicity 3
Species Sensitivity Distribution (SSD) 3
3. Results 4
Comparative Toxicity Summary 4
Acute Toxicity 4
Chronic Toxicity 5
Estimating Toxicity 5
Acute Toxicity 5
Acute Estimate Accuracy and Uncertainty Analyses 5
Chronic Toxicity 5
Species Sensitivity Distribution (SSD) 5
Acute Toxicity SSD 5
Chronic Toxicity SSD 6
4. Discussion 7
111
-------
NOTICE
The U.S. Environmental Protection Agency through its Offices of Research and Development, Pesticide
Programs, Pollution Prevention and Toxics, and Water partially funded and cooperated in the research
described herein under EPA project Nos. DW14935155, DW14936559, and DW14939002 (U.S. Geological
Survey, Biological Resources Division, Columbia Environmental Research Center, Columbia, MO) and
No. CR82827901 (University of Missouri, College of Agriculture, Food and Natural Resources, Agricultural
Experiment Station-Statistics, Columbia, MO). It has been subjected to the Agency's peer and administrative
reviews and has been approved for publication as an EPA document. Mention of trade names or commercial
products does not constitute endorsement or recommendation for use.
IV
-------
ABSTRACT
Many times, endangered, threatened, and candidate endangered species (collectively known as listed
species) have been thought to be uniquely sensitive to chemicals. The purpose of this cooperative
research effort (U.S. Environmental Protection Agency, U.S. Geological Survey, U.S. Fish and Wildlife
Service, University of Missouri) was to determine: 1) if listed aquatic vertebrate species are more sensitive
to chemicals than non-listed species; 2) if common surrogate test species represent listed species
toxicologically; and 3) if predictive acute and chronic models can be applied to hazard assessments with
listed species where direct toxicity testing is not prudent or impractical. Toxicity tests were conducted with
29 species of fishes and amphibians (endangered species and a set of surrogates) and five chemicals
(carbaryl, copper, 4-nonylphenol, pentachlorophenol, and permethrin) representing a broad range of toxic
modes of action. For acute toxicity, rainbow trout (Oncorhynchus mykiss), the most sensitive surrogate
species, was equal to or more sensitive than listed and related aquatic vertebrate species 80% of the
time. Only 3% of the species were significantly (P< 0.05) more sensitive than rainbow trout, and even
then, the differences were within or very close to a factor of two (normal intra- and interlaboratory variation
= 2-5x). Under similar environmental conditions, chronic toxicity tests with copper and pentachlorophenol
indicated no significant greater sensitivity between rainbow trout and the listed species, spotfin chub
(Cyprinella monacha) and fountain darter (Etheostoma fonticola). Using Interspecies correlation
estimation (ICE) for estimating acute toxicity, 100% of the values for listed or related species were within
or very close to a factor of two of the observed values (n = 70, mean = 1.1, range = 0.49 - 2.2). Acute-to-
chronic (ACE) estimated chronic toxicity values were within a factor of two of observed values 80-90% of
the time and 100% within a factor of three. Species sensitivity distributions (SSD) were also developed to
determine the 5th percentile effect among observed data and different sets of estimated data. The most
accurate estimated acute toxicity SSDs were in using the surrogate species having the best correlation
model in ICE (SSD = 0.95x observed SSD; range = 0.88 - 0.98). SSDs for chronic toxicity were also quite
good with ACE-estimated chronic data or ICE-estimated acute data/acute-chronic ratio. 5th percentile
estimates averaged 0.95 times those for observed data (range = 0.46 -1.3). The results suggest that
listed aquatic vertebrate species are not universally more sensitive to contaminant exposure than other
aquatic vertebrate species on a toxicological basis. Surrogate test species do appear to represent listed
species toxicologically, at least for aquatic vertebrates, and toxicities and hazard assessments (SSD) can
be estimated accurately and precisely, not only for listed species, but other species with little or no toxicity
data as well.
-------
FIGURES
Page
Figure 1a Species sensitivity (static acute toxicity) distribution for carbaryl 9
Figure 1b Species sensitivity (static acute toxicity) distribution for copper 10
Figure 1c Species sensitivity (static acute toxicity) distribution for 4-nonyiphenol 10
Figure 1d Species sensitivity (static acute toxicity) distribution for pentachlorophenol 11
Figure 1e Species sensitivity (static acute toxicity) distribution for permethrin 11
Figure 2. Sensitivity of darters relative to other aquatic species 12
Figure 3a Species sensitivity distributions for observed acute toxicity data and
ICE-based estimated data using the surrogate species (fathead minnow
rainbow trout, or sheepshead minnow) having the best correlation with
the respective endangered species 13
Figure 3b Species sensitivity distributions for observed acute toxicity data and
ICE-based estimated data using fathead minnow as the only surrogate
species 14
Figure 3c Species sensitivity distributions for observed acute toxicity data and
ICE-based estimated data using ECOSAR fish values as the only
surrogate species 15
Figure 4 Test species required to include most sensitive species for acute toxicity
tests 16
Figure 5 Range of effects and geometric mean of variables on acute toxicity of
chemicals to aquatic organisms 16
VI
-------
TABLES
Table 1. Surrogate and endangered species tested with carbaryl, copper, 4-nonylphenol,
pentachlorophenol, and permethrin 17
Table 2. Acute static toxicity of carbaryl to surrogate test species and endangered
species 18
Table 3. Acute static toxicity of copper and 4-nonylphenol to surrogate test
species and endangered species 19
Table 4. Acute static toxicity of pentachlorophenol and permethrin to surrogate
test species and endangered species 20
Tables. Acute toxicity sensitivity rankings by 96-h LC50 21
Table 6. Chronic toxicity (ug/L) of copper (50 mg/L hardness) and
pentachlorophenol (6.5 pH) to fathead minnow, spotfin chub, rainbow
and fountain darter. 22
Table 7. Interspecies correlations (X2 = a + bX,) for all species with fathead
minnow as the surrogate species 23
Table 8. Interspecies correlations (X2 = a + bX^ for all species with rainbow trout
as the surrogate species 24
Table 9. Interspecies correlations (X2 = a + bX,) for all species with sheepshead
minnow as the surrogate species 25
Table 10. Correlation coefficients (r) and number of paired tests for all species and
chemicals analyzed for interspecies correlations 26
Table 11. Observed and estimated 96-h LC50s (ug/L) for five chemicals and 15
aquatic vertebrates using interspecies correlation analysis (ICE) and
surrogate species (fathead minnow, rainbow trout, or sheepshead
minnow) acute toxicity values 27
Table 12. Observed and estimated (ACE) chronic toxicities (ug/L) of copper (50
mg/L hardness)and pentachlorophenol (6.5 pH) for fathead minnow,
spotfin chub, rainbow trout, and fountain darter. 28
Table 13. Calculated 5th percentile values for observed and estimated acute
toxicity data (96-h LC50s in ug/L) using three methods 29
Table 14. Calculated 5th percentile values (ug/L) for observed and estimated
chronic toxicity data using three methods 30
vn
-------
ACKNOWLEDGMENTS
This project was sponsored in part by the U.S. Environmental Protection Agencys Office of Research
and Development, Pesticide Programs, Pollution Prevention and Toxics, and Water under Cooperative
Agreements DW14935155, DW14936559, DW14937809, DW14939002, and CR82827901; New York
Department of Environmental Conservation; and the U.S. Fish and Wildlife Service. We thank the federal,
state, and private hatcheries that provided the species used in testing, as well as E. Greer for culturing
test organisms. Technical assistance was provided by W. Brumbaugh, E. Brunson, K. Feltz, J. Folse, D.
Hardesty, C. Henke, S. Holbrook, C. Ivey, J. Kunz, T. May, C. Orazio, M. Tanner, N. Wang, D. Whites, and
R. Wiedmeyer; technical suggestions and support by T. Augspurger, K. Hattala, C. Ingersoll, D.R. Mount,
and G. Neuderfer; data analysis by A. Asfaw, V. Engle, and S. Raimondo; and graphics by V. Camargo and
S. Embry. We also thank Valerie Coseo for manuscript preparation. Peer reviews were contributed by T.
Augspurger, T. Bailey, K. Dickson, C. Flaherty, H. Galavotti, T. Henry, J. Hyland, T. Linton, D. Randall, M.
Reiley, D. Rodier, T. Steeger, and T. Waller.
Vlll
-------
1. INTRODUCTION
The widespread use of pesticides and other
commercial chemicals potentially poses a risk to
threatened and endangered species. Because
by definition, the distribution of each species is
limited, additional adverse stresses on these
populations could lead to extinction. In addressing
the risk of chemicals to endangered species, the
U.S. Environmental Protection Agency (EPA)
is the primary federal agency that regulates
chemical substances in the environment of
the United States. This authority is granted
primarily within three statutes: the Federal
Insecticide, Fungicide, and Rodenticide Act
(FIFRA; FL80-140), the Toxic Substance Control
Act (TSCA; PL94-469), and the Clean Water Act
(CWA; Section 101 (a) (3)). The FIFRA is used
to regulate pesticides that are manufactured
specifically for their toxicity and are intended for
direct application to the environment. The TSCA
regulates the production, use, transportation, and
disposal of chemicals of commerce, excluding
pesticides. The CWA prohibits the discharge
of pollutants in toxic amounts to water bodies
of the United States. The Endangered Species
Act of 1973, affords additional environmental
protection through Section 7 consultations,
requiring Federal agencies to ensure that any
action authorized, funded, or carried out by them
is not likely to jeopardize the continued existence
of endangered species or modify their critical
habitat. The chemical registration and regulation
responsibilities of EPA fall under the Endangered
Species Act.
Toxicity testing under FIFRA may require four
categories of data, including acute toxicity tests
with freshwater, estuarine, and marine fish
and invertebrates; embryo-larval and life-cycle
studies with fish and invertebrates; chemical
residue studies; and field testing. In the absence
of valid test data, TSCA risk assessments are
generally based on quantitative structure activity
relationships (QSAR). An integrated community-
based statistical approach is used to derive
water quality criteria, recommended under the
CWA, with a minimum multispecies data base
(Stephan et al. 1985) to protect aquatic organisms
from unacceptable adverse effects. As part of the
National Pollutant Discharge Elimination System
permit process, protection of aquatic environ-
ments from toxic discharges commonly includes
whole effluent toxicity tests with Ceriodaphnia
dubia, fathead minnow, and the alga, Selenastrum
capricornutum (U.S. EPA 1991).
The selection of surrogate test species used in
aquatic toxicity testing is critical to regulatory
processes because of the need to be predictive of
a large number of species and their sensitivities,
including endangered species. The test species
used for toxicity assessments are representative
and generally protective of other species, including
those that are threatened or endangered. Mayer
and Ellersieck (1986) compiled an acute toxicity
data base for 410 chemicals and 66 species of
freshwater animals and reached three conclusions:
1) for a given chemical, acute toxicity among species
ranged over five orders of magnitude; 2) for a given
species, acute toxicity among chemicals ranged over
nine orders of magnitude; and 3) no single species
was always the most sensitive to all chemicals, but
a group of surrogate species, as used in U.S. EPA
programs, did include the most sensitive species
toxicity value most of the time.
Since the advent of the Endangered Species
Act, endangered species have sometimes been
assumed to be more sensitive to chemicals and
other stressors than non-endangered species. A
more frequent concern is that sensitivity is unknown,
thus sensitivity estimates should include margins of
safety to avoid consequences of incorrect decisions
for rare taxa. Under FIFRA, a margin of safety
is added to risk assessment levels of concern
for endangered species. With acute toxicity for
example, it is presumed that minimal risk will occur
to endangered aquatic organisms from pesticides
(Urban and Cook 1986) if:
• EEC (Estimated environmental
concentration)/LC10 < 0.1 (when a
slope is available).
-------
• EEC/LC50 < 0.05 (when no slope is
available).
• EEC < lowest chronic no-effect
concentration.
TSCA does not provide the legislative authority to
require extensive testing, a priori, as does FIFRA.
The risk assessment process, many times, depends
upon quantitative structure activity estimates for
toxicity. With CWA, it is generally assumed that
derived water quality criteria are protective of all
aquatic species. A reasonable level of protection
(95%) will be provided if a diverse number of genera
are used in development of the criteria. In addition,
a margin of safety is added by limiting exceedances
of the Criteria Maximum Concentration (CMC, acute
toxicity) or the Criteria Continuous Concentration
(CCC, chronic toxicity) values for 1 hour or 4 days,
respectively, once every 3 years; ammonia criterion
includes a longer averaging period. However,
there is no limit on the exceedance in terms of
concentration. In order to meet the concentration/
frequency criteria from an engineering perspective,
a treatment plant must be designed with a goal of
keeping concentrations below the criteria, but actual
discharge monitoring is not set up to capture the
frequency exceedance criteria.
Under current regulations, unnecessary margins of
safety may be applied for endangered species, if
their sensitivity to toxic chemicals is not understood.
Thus, a major question remains as to whether
endangered species may be more sensitive to
chemical stressors than other species. More
important, however, in the absence of toxicity data
for endangered species, what is a reasonable
protective estimate of the toxicity? To address these
questions, at least in part, the objectives of this
effort were to:
• Summarize the toxicity data,
• Assess whether surrogate aquatic test
test species are toxicoiogically
representative of endangered aquatic
vertebrate species,
• Evaluate the application of acute and
chronic estimation models for endangered
aquatic vertebrate species, and
• Establish species sensitivity distributions
(SSDs) for surrogate and endangered
aquatic vertebrate species, contrasting
5th percentile values of actual and
estimated data.
2. METHODS
2,1 Toxicity Tests
Static acute toxicity tests were conducted on
several species (Table 1) in basic accordance with
procedures described in ASTM (2003a). Methods
for comparative toxicity are provided only briefly
since specific methods have been published
elsewhere (Besser et al. 2001, 2005; Bridges
et al. 2002; Dwyer et al. 1995, 1999a,b, 2000,
2005a,b; and Sappington et al. 2001). Additional
data (unpublished, C.M. Bridges, personal
communication) for larval amphibians and carbaryl
are also presented. Acute exposures for most
species included five chemicals representing a
broad range of modes of action:
• Carbaryl - cholinesterase inhibitor
• Copper - osmoregulation
interference
• 4-Nonylphenol - narcotic and
oxidative stressor
• Pentachlorophenol - oxidative
phosphorylation uncoupler
• Permethrin - sodium channel blocker
Early life-stage toxicity tests were conducted
in general accordance with ASTM (2003b).
Chronic exposures included only copper and
pentachiorophenol tested with fathead minnow,
spotfin chub, rainbow trout, and fountain darter.
For both acute and chronic tests, the copper
toxicity values were adjusted to represent toxicity
at 50 mg/L hardness (U.S. EPA 1985) and
pentachiorophenol values adjusted to pH 6.5
(U.S. EPA 1986) except for Cyprinodontidae. Both
surrogate and endangered fishes and amphibians
were tested (Table 1).
2.2 Statistical Analyses
For the comparative toxicity summary, the data
of Besser et al. 2001, 2005; Bridges et al. 2002;
Dwyer et al. 1995, 1999a,b, 2000, 2005a.,b; and
Sappington et al. 2001 were used, but were
statistically re-analyzed for consistency or further
analyzed.
2.3 Acute Toxicity
LC50 Estimation. All raw data from our previous
publications were re-analyzed; 96-h LCSOs and
95% confidence limits (CL) were derived by probit
-------
analysis (Finney 1978; PROC PROBIT-SAS/
STAT, SAS Institute Inc. 2004). In a few tests, the
LC50 was determined by linear interpolation, and
in 14 tests, the 96-h LC50 was derived by probit
analysis, but the respective 95% CL had to be
estimated using Spearman-Karber techniques
(Hamilton et al. 1977a, b). For some tests, CL
could not be estimated.
Comparative Toxicity. The 96-h LC50 data
were analyzed by one-way analysis of variance
(Snedecor and Cochran 1989) to compare
species sensitivity within a chemical. Log10
transformation of toxicity data was used and
met the assumptions for normally distributed
data in analysis of variance. The design was
not balanced because the numbers of tests per
species for each chemical were unequal (number
of tests per species/chemical combination =
1 -6; replicates per test = 2-3). To determine
mean differences among LCSO's, Fisher's least
significant difference (LSD) was used (PROC
GLM-SAS/STAT, SAS Institute Inc. 2004). In
addition, sum of ranks/n analyses, based on the
Iog10 of the values or means, were also conducted.
2.4 Chronic Toxicity
Log10 chronic values for survival (arcsine
transormed) and growth were determined by
analysis of variance. Log10 chronic values were
then analyzed by an unbalanced n one-way
analysis of variance (Snedecor and Cochran
1989), followed by Fisher's LSD, to compare
species within a chemical by effect (PROC GLM-
SAS/STAT Institute Inc. 2004).
2.5 Estimating Toxicity
Acute toxicity estimation. Interspecies correlation,
Model II least squares methodology, (both
variables are independent and subject to
measurement error (Snedecor and Cochran
1989), was used to develop correlation models
for the surrogate test species (fathead minnow,
rainbow trout, sheepshead minnow) with all
other species tested. Slopes and intercepts
were derived from the equation Iog10 X2 = a +
b(log10X1), where X2= the estimated 96-h LC50
for the species in question and X1 = the known
96-h LC50 value for one of the three surrogate
test species. When more than one LC50 value
existed for a species/chemical combination, the
geometric mean was used. A data set of correlation
parameters was established for these acute toxicity
data and estimates made with the ICE (Interspecies
Correlation Estimation) software (Asfaw et al. 2003).
Accuracy of 96-h LC50 estimates for endangered
and related species was determined by deriving
a ratio (estimated value/actual value), followed by
univariate analysis of the ratios (PROC Univariate-
SAS/STAT, SAS Institute Inc. 2004).
Chronic toxicity estimation. Chronic toxicity values
of copper and pentachlorophenol were estimated
for fathead minnow, spotfin chub, rainbow trout,
and fountain darter using raw acute toxicity data
(mortality observations within each exposure
concentration time and at each observation time
of 24, 48, 72, and 96 h) for the respective species.
Fountain darter and spotfin chub were selected
because of availability and known culture techniques.
The Accelerated Life Testing or Linear Regression
Analysis models, within the Acute-to-chronic
estimation (ACE) v2.0 software (Ellersieck et al.
2003, Mayer et al. 1999, 2002), were used for the
estimations. Estimation accuracy was determined
by comparing predicted chronic values to actual
chronic values (geometric mean of no-observed-
effect [NOEC] and lowest-observed-effect [LOEC]
concentrations -r by observed values).
2.6 Species Sensitivity Distribution (SSD)
The assessment of hazard was accomplished by
comparing toxicity responses of endangered to
surrogate test species (fathead minnow, rainbow
trout, sheepshead minnow) and by comparing the 5th
percentile of species sensitivity distributions (SSD)
to listed species toxicity values. For estimating the
5th percentile of species sensitivity distributions,
three methods were used and compared (described
below). In addition, the 5th percentile estimations
were conducted for actual acute and chronic values,
values estimated by ICE (Asfaw et al. 2003) and
QSAR (quantitative structure-activity relationships)
for acute toxicity, and values estimated by ACE
(Ellersieck et al. 2003) and acute-chronic ratios
(ACR) for chronic toxicity.
Graphic species sensitivity distributions were
derived using the methods of Dyer et al. (2006).
The hazardous concentration protective of 95% of
species (5th percentile value or HC5) for all SSDs
was determined by the methods of Solomon et al.
(1996, 2000), Aldenberg and Jaworska (2000), and
Stephan et al. (1985). The acute toxicity data sets
-------
that were used included: 1) observed data, 2)
data derived by ICE (Asfaw et al. 2003) using
the surrogate species (fathead minnow, rainbow
trout, or sheepshead minnow) having the best
correlation statistics, 3) data derived by ICE using
only the surrogate fathead minnow value, 4) data
derived by ICE using the ECOSAR (Ecological
Structure Activity Relationships) fathead minnow
value (U.S. EPA 2001), or 5) data derived by
ICE using the ASTER (Assessment Tools for
the Evaluation of Risk) fathead minnow value
(Russom 1991, 1997). The chronic data sets
used included: 1) observed data from criteria
documents (U.S. EPA 1985, 1986), 2) observed
data from criteria documents plus observed
data for fountain darter and spotfin chub, 3)
observed data from criteria documents plus ACE
(Ellersieck et al. 2003) estimated chronic values
for fountain darter and spotfin chub, 4) observed
data from criteria documents plus ICE (Asfaw
et al., 2003) estimated acute values divided by
the final criterion ACR (USEPA 1985, 1986) for
estimated chronic values for fountain darter and
spotfin chub, or 5) observed data for Daphnia
tnagna and fathead minnow with estimated
values for all other species (acute ICE estimates
divided by the ACR).
3. RESULTS
3.1 Comparative Toxicity Summary
3.1.1 Acute Toxicity
Control survival, with and without solvent, was
always greater than 90% for all species except for
Atlantic and shovelnose sturgeons (< 90%). For
these two species, control survival appeared to
be affected by the acetone carrier and only acute
toxicity results for copper (no solvent) with Atlantic
and shovelnose sturgeon are presented. Control
survival for fountain darter was acceptable, but
mortality (5-15%) occurred consistently among
all chemicals tested. Ten percent or less is
considered acceptable (ASTM 2003). These
observations indicate that, any conclusions
regarding the acute chemical sensitivity of
sturgeon and fountain darters should be regarded
with caution until testing methodology has been
refined. See Dwyer et al. (1995; 1999a,b; 2000;
2005a).
Overall, species of the families Acipenseridae,
Salmonidae, and Percidae were more sensitive
to the five chemicals than species in Cyprinidae,
Catostomidae, Cyprinodontidae, Poecillidae,
Bufonidae, and Ranidae (Tables 2-4, Fig. 1a -1e).
Differences between listed species and the suite
of surrogates were generally less than two-fold.
Permethrin was the most toxic compound and
carbaryl was the least toxic. The LC50s for rainbow
trout (Salmonidae) were always lower than the
LC50s for fathead (Cyprinidae) and sheepshead
minnows (Cyprinodontidae).
For carbaryl, 96-h LC50s ranged from 1435
ug/L (Apache trout) to 12,303 ug/L (boreal toad).
Amphibians were the least sensitive to carbaryl
(4592-12,303 ug/L) with all values being in the upper
50th percentile. Copper LC50s ranged from 18 ug/L
(fountain darter) to 1306 ug/L (Leon Springs pupfish).
Toxicity results with 4-nonyphenol ranged from 81
(shortnose sturgeon) to 553 ug/L (Leon Springs
pupfish). Pentachlorophenol LCSOs ranged from
11 (shortnose sturgeon) to 191 ug/L (sheepshead
minnow). Permethrin toxicity ranged from 1.7 (spotfin
chub and Lahontan cutthroat trout) to 39 ug/L
(bonytail chub).
Based on 96-h LC50s, rainbow trout had a summary
sensitivity ranking across the five chemicals of 5.2
(Table 5). Only 5 of 23 species fell below that value,
and of those species lower LCSOs, the average
factor was 0.7 (± 0.2 SD) that for rainbow trout. The
summary ranking for fathead minnow across the
five chemicals was 13.2 (range of species rankings
- 2.0 to >18) and was one of the more tolerant
species tested overall. The results indicate that the
acute sensitivities of rainbow trout were equal to or
more sensitive than that for listed and other species
80% of the time. However, rainbow trout acute
toxicity values were equal to or within a factor of 2
of the most sensitive species 97% of the time; the 3
values exceeding a factor of 2 were: 4-nonylphenol
- shovelnose sturgeon (0.42x) and fountain darter
(0.47x); pentachlorophenol - shortnose sturgeon
(0.44x) - - the only values that were significantly (P<
0.05) more sensitive than those for rainbow trout.
The ranking (Table 5) is in general agreement with
that previously derived by Dwyer et al. (2005).
As mentioned previously, the data for the fountain
darter, as well as sturgeons should be used with
caution. Further, the fountain darters in the present
study were very sensitive to copper. Five other
darter species were 3.8 -7.9 times more tolerant (Fig.
2, U.S. EPA 1985). Additional research including
culture techniques used in acute toxicity testing is
-------
needed to determine if sturgeons and the fountain
darter are truly more sensitive to chemicals than
other species.
3.1.2 Chronic Toxicity
For copper, the fountain darter tended to be the
most sensitive species followed by rainbow trout,
fathead minnow, and spotfin chub. Although no
statistically significant differences (P < 0.05) were
found (Table 6), the sample size is small. Growth
was the slightly more sensitive endpoint overall
for both copper and pentachlorophenol. Fathead
minnow was the most tolerant species tested with
pentachlorophenol. Fountain darter was the most
sensitive species, but the data for fountain darter
were not significantly different from that for spotfin
chub and rainbow trout. See Besser et al (2001,
2005).
3.2 Estimating Toxicity
3.2.1 Acute Toxicity
ICE (Asfaw et al. 2003) models were developed
for all species tested using the fathead minnow,
rainbow trout, or sheepshead minnow as the
surrogate species (Tables 7-9). Correlations
were quite strong (r) and slopes (b) were close to
1.0 in most cases. Overall, the fathead minnow
and rainbow trout were the best surrogate
species representing other species. Correlation
coefficients (r) were greater than 0.910 using
rainbow trout as the surrogate (Table 8), 0.882
for fathead minnow (Table 7), and 0.875 for
sheepshead minnow (Table 9). No significant
slopes existed for boreal toad and the three
surrogate fishes. However, the surrogate species
used should be as close as possible taxonomically
to the species being estimated (Asfaw et al. 2003).
The slope (1.090) is significant (p = 0.04) when
using southern leopard frog as the surrogate for
boreal toad. Correlations were also conducted
using each species tested as a surrogate (Table
10). Most correlations were high (r > 0.90), with
lower r values usually being associated with the
surrogate and other species not being from the
same taxonomic family.
3.2.2 Acute Estimate Accuracy and
Uncertainty Analyses
Of the 70 estimated acute toxicity values, 96%
were within a factor of 2.0 (> 0.5 < 2.0) of the
observed values (Table 11). Three ratios, or 4%,
were only marginally greater than a factor of 2.0
(0.49, 2.1, 2.2). Univariate statistics of the ratios
of the estimated acute toxicity value divided by the
observed acute value (> 1.0, estimate higher than
observed; < 1.0, estimate lower than observed)
were:
Parameter
Value
Number of observations
Mean
95% confidence limits
Standard deviation
Standard error
Median
1 st - 99st percentile
5th - 95th percentile
10th-90th percentile
25th - 75th percentile
Range
70
1.06
0.98- 1.14
0.35
0.041
1.00
0.49 - 2.24
0.60-1.61
0.72-1.54
0.79-1.30
0.49 - 2.24
Previous interlaboratory and intralaboratory acute
toxicity test comparisons (DeGraeve et al.1991,
Lemke 1981, Schimmel 1981) demonstrated
that a two-to five-fold difference (i.e., highest/
lowest acute toxicity values) in LC50s existed for
the same species/chemical combinations and
methodology. Intralaboratory test comparisons
in the same sources, indicated that LCSOs
seldom varied by more than a factor of 2.0.
Thus, the differences between observed and
estimated acute toxicity values with the species
examined are within, or are very close to, normal
intralaboratory variation.
3.3 Chronic Toxicity
Use of ACE (Ellersieck et al. 2003) proved highly
accurate as an estimator of chronic toxicity (Table
12). Significant differences among ACE-estimated
values were not determined since each estimate
is a single value. Dividing the ACE estimated
value by the lowest Chronic Value, provided an
overall ratio of 0.98 (median, 1.1) or approximately
the same as the actual value. Regardless of the
analyses, 80 percent of the estimates were within
a factor of two of the observed values and two
estimates (0.33, 0.37) were still within normal
intra- and interlaboratory variation (2-5x).
3.4 Species Sensitivity Distributions (SSD)
3.4.1 Acute Toxicity SSD
-------
Three frequently used SSD methods (Aldenberg
and Jaworska, 2000; Solomon et al. 1996, 2000;
Stephan et al. 1985) were applied to calculate 5th
percentile values for acute toxicity of the chemicals
to endangered vertebrates. This, by no means,
indicates a rigorous comparison of the methods
because the data sets are small (n = 15-18).
Wheeler et al. (2002) have previously assessed
different SSD statistical models for determining 5th
percentile values and found differences. Various
SSD models have been suggested (log-normal, log-
logistic, log-triangular and bootstrap techniques) as
well as using censored data sets for estimating the
5th percentile, but a consensus does not seem to
exist (Posthuma et al. 2002). However, we conducted
these analyses to determine potential differences
between observed and estimated values among SSD
methodologies for endangered species. Examples
of SSDs for permethrin, comparing observed data
and three estimated data methods, and the model of
Aldenberg and Jaworska (2000) indicate that use of
estimated data leads to a more
conservative lower portion of the curve and a less
conservative upper portion (Fig. 3a-c).
For acute data (Table 13), SSDs were conducted with
three models (Solomon et al. 1996, 2000; Aldenberg
and Jaworska 2000, and Stephan et al. 1985) using
five data sets; 1) observed data, 2) ICE estimated
data for several tested species with observed values
using the best correlation among three surrogate
species (fathead minnow, rainbow trout, sheepshead
minnow), 3) ICE estimated data using fathead
minnow only as the surrogate, 4) ECOSAR estimated
fathead minnow data, and 5) ASTER estimated
fathead minnow data. The best estimated data, as
compared to observed data, to determine the 5th
percentile was obtained using the surrogate species
having the best correlation model in ICE. The 5th
percentile estimates were slightly conservative and
averaged 0.94x (median = 0.92; SD = 0.08; range
= 0.81 - 1.1 x) that of observed data. Using fathead
minnow data only resulted in a less conservative
mean factor of 1.2x (median = 0.99; SD = 0.54; range
= 0.64 - 2.7x). Fathead minnow values determined
with QSAR (ECOSAR, ASTER) resulted in even less
conservative 5th percentile estimates, but were all
close to or within an order of magnitude of observed
values (ECOSAR and ASTER - mean factor of 3.6x
[range = 0.27 - 12x], ECOSAR - mean factor of
4.4x [range = 0.43 - 12x], ASTER - mean factor of
2.2x [range = 0.27- 4.1x]). ASTER values resulted
in more accurate and less variable 5th percentile
estimates than ECOSAR; however, the data sets
were too small to make a good comparison
between the two QSAR estimate techniques.
The 96-h LC50 of the most sensitive species
tested (Tables 2- 4) was always greater than or
very close to the 5th percentile values generated
by the methods of Solomon et al. (1996, 2000)
and Stephan et al. (1985) (5th percentile mean
= 0.72x, range = 0.65 - 1.1 the most sensitive
species LC50 tested). The 5th percentile values
generated by the Aldenberg and Jaworska (2000)
method, regardless of observed or estimated data,
were always greater than the 96-h LC50 for the
most sensitive species tested (mean = 2.6x, range
3.4.2 Chronic Toxicity SSD
For chronic data (Table 14), SSDs were conducted
with the same three models as for acute data
and four data sets; 1 ) observed data from the
respective criteria document plus data for spotfin
chub and fountain darter, 2) observed data from
criteria document plus chronic data estimated
by ACE for spotfin chub and fountain darter, 3)
observed data from criteria document plus spotfin
chub and fountain darter data derived by ICE
estimated acute values divided by the ACR, and
4) observed data for Daphnia magna and fathead
minnow from criteria document plus all other
species estimated by ICE divided by the ACR. All
three data sets containing data estimated by ICE
or ACE were highly accurate when compared
to the 5th percentile of the observed data set,
except for the SSD method of Stephan et al.
(1985). Using Solomon et al. (1996, 2000) and
Aldenberg and Jaworska (2000), the first two data
sets containing estimated data (ACE and ICE for
the two endangered species) averaged the same
as the 5th percentile for observed data (ACE -
factor of 1 .Ox, range = 0.83 - 1 . 1 x, ICE - factor of
1.0x, range = 0.94 - 1.1x). The data set having
observed data for Daphnia magna and fathead
minnow with ICE-estimated data for all other
species produced a 5th percentile value that was
also very close to observed data, but was more
conservative (average factor of 0.87x, range = 0.52
- 1 .3x) and more variable.
For observed copper data, the most sensitive
species chronic value (fountain darter, 2.7 ug/L)
was slightly greater than the 5th percentile value
(2.3 (jg/L) using Solomon et al. (1996, 2000) and
Stephan et al. (1985), but was less than half (0.4x)
-------
the SSD generated by Aldenberg and Jaworska
(2000). The 5th percentile values for data sets
containing estimated data were also less than the
chronic value for fountain darter, ranging from 0.45
to 2.4 ug/L, with the exception of Aldenberg and
Jaworska (2000), which ranged from 3.2 to 6.8
ug/L. With pentachlorophenol, all 5th percentile
values derived with Aldenberg and Jaworska
(2000) were about two times greater than the
chronic value for fountain darter (3.9 ug/L), the
most sensitive species. The Solomon et al. (1996,
2000) technique resulted in somewhat consistent
5th percentile values that were slightly less than
the fountain darter chronic value. The method of
Stephan et al. (1985) resulted in highly variable
5th percentile values (0.16-1.2 ug/L) well below
the fountain darter chronic value.
Various approaches in estimating chronic toxicity
data were also conducive to estimating 5th
percentile SSDs that corresponded well to SSDs
for observed data. Although small acute and
chronic data sets were used, the SSD method of
Solomon et al. (1996, 2000) was conservative and
produced the most consistent SSDs for including
the most sensitive species. It may be that the
Solomon methodology is best when using small
data sets.
4. DISCUSSION
Overall, endangered fishes and amphibians do
not appear to be significantly more sensitive to
chemicals than non-endangered species, based
on the tests performed here. This conclusion is
consistent to that reached by Hansen et al. (2002)
and Fairchild et al. (2005) who also indicated
that threatened and endangered species are
not uniquely sensitive to toxicants. Also, the
surrogate test species evaluated were found to
be toxicologically representative of endangered
species. Rainbow trout are especially useful when
estimating acute toxicity for sensitive organisms;
rainbow trout LCSOs were < those of sensitive
species 78% of the time but were within a factor
of two, 97% of the time. The three exceptions
had factors of 0.42 - 0.47x those of rainbow trout
and thus were very close to a factor of 2 (or
0.5). Dwyer et al. (2005a) found that a factor of
0.63 could be applied to a rainbow trout LC50
to estimate an endangered or other fish species
LC50. He further derived a conservative factor of
0.46 (0.63-1 SD) and a highly conservative factor
of 0.33 (0.63-2 SD). It must also be kept in mind that
previous inter- and intralaboratory acute toxicity test
comparisons (DeGraeve et al. 1991, Lemke 1981,
Schirrtmel 1981) demonstrated that a two-to five-fold
difference (i.e., highest/lowest acute toxicity values)
in LC50s existed for the same species/chemical
combinations and methodology. Intralaboratory test
comparisons indicated that LCSOs seldom vary by
more than a factor of 2.
Muffimert et al. (2003) reported ammonia LC50s
for mussels were 0.6 to 0.9 times LCSOs of rainbow
trout. In another study on effluent toxicity, Dwyer
et al. (2005b) indicated that endangered fishes
were more sensitive than fathead minnows 19%
of the time (0.2 - 0.9x) and more sensitive than
Ceriodaphnia dubia only 4% of the time (0.6 - 0.7x).
Besser et al. (2005) also found that chronic values for
two endangered fishes (fountain darter and spotfin
chub) were approximately O.Sx that of rainbow trout
or greater. The significance of this is that a factor of
0.5 times the more sensitive surrogate test species
(i.e., rainbow trout or Ceriodaphnia dubia) appears to
encompass LCSOs, chronic values, or effluent IC25s
of the more sensitive endangered species.
The EPA guidance (U.S. EPA 1978, Stephan et
al. 1985) for determining a no- or low-acute-effect
concentration (Continuous Maximum Concentration,
CMC) requires multiplying the Final Acute Value
by 0.5. Dwyer et al. (2005a) determined that
the factor was 0.56 for the endangered fishes
studied. However, in other studies with ammonia
and freshwater mussels, toxicity to mussels was
approximately 0.1-0.6x (Augspurger et al. 2003) and
0.2-1.2x (Wang et al. 2007) that for the common
surrogate fish species tested, rainbow trout and
fathead minnow, respectively. Therefore, derived
safety factors (e.g., 0.5x) within a taxa (e.g., fishes)
may not be applicable to other taxa and specific
chemicals (i.e., mussels and ammonia), and thus
may not be protective.
Surrogate test fishes do appear to represent
endangered fishes, in both toxicity and sensitivity.
Although Mayer and Ellersieck (1986) stated that
"no single species was always the most sensitive
to all chemicals," they further reported that, in a
comparison of 40 chemicals, the most sensitive
species could be identified using a combination
of surrogate test species (i.e., either Daphnia or
rainbow trout were the most sensitive 93% of the
time; in the other 7%, fathead minnow or bluegill
-------
were the most sensitive - - all surrogate test
species). A further analysis of random surrogate
species toxicological representation of other
species among all chemicals (data from Mayer
and Ellersieck, 1986) indicated that a suite of
three test surrogates (daphnids, rainbow trout, and
gammarids) could account for the most sensitive
species 87.5% of the time (Fig. 4); additional test
species contributed only 2.5% each to identifying
the most sensitive species.
The major acute and chronic toxicity testing
programs for aquatic species that existed in
government and industry basically ended in
the mid 1980s, and no major toxicity testing
programs are anticipated in the near future. Thus,
methods for probability-based toxicology and risk
assessment are a priority, including predictive
toxicology and probability-based risk assessment
for endangered species -- all species cannot
practically be tested with all chemicals. The
greatest range of uncertainty in acute toxicity
tests is due to species within a chemical and
chemicals within a species (Fig. 5). With the data
of Mayer and Ellersieck (1986), it was found that
acute toxicity data for species within a chemical
ranged as high as 5 orders of magnitude and
chemicals within a species, ranged up to 9 orders
of magnitude. The uncertainty of acute toxicity
estimates for chemicals within a species should
be addressed with quantitative structure activity
relationships (QSAR), although QSAR is generally
limited to three organisms (fathead minnow,
daphnid, and an alga). ICE and ACE were
developed to address the uncertainty of species
within a chemical.
Acute and chronic estimation models (ICE,
A-sfaw et al. 2003; ACE, Ellersieck et al. 2003)
were shown to be both highly accurate and
precise in estimating acute and chronic toxicity to
endangered species. Of 70 ICE-estimated acute
toxicity values, 96% were within a factor of 2.0
(range = 0.49-2.2) of the observed values. The
mean factor was 1.1 (SD - 0.35) with a median
of 1.0 (>1.0, estimate higher than observed; <1.0,
estimate lower than observed). ACE-estimated
chronic values were also very close to observed
values. The mean factor was 0.9 (range = 0.33 -
1.6), with two of ten factors being slightly greater
than a factor of 2.0 (0.33 and 0.37). Mayer et al.
(2002) also found a very high degree of accuracy
in predicting chronic toxicity from acute toxicity
data using time-concentration-effect models
(ACE v1.0, Mayer et al. 1999) with 7 fish species
and 18 chemicals. The significance of the ICE and
ACE estimations is that they are all within normal
intra- and interlaboratory variation (2-5x), and thus
acute and chronic values for endangered fish and
other aquatic vertebrate species can be accurately
estimated when toxicity values do not exist. If a
correlation for a specific endangered species does
not exist in ICE, the genus or family value of the
endangered species can be estimated, although
the estimated genus or family values may have
more uncertainty than an estimated species value.
Also, the method of Dwyer et al. (2005a) could be
used; i.e., 0.5x the rainbow trout acute and chronic
values or Ceriodaphnia dubia effluent values. If an
acute toxicity value does not exist for rainbow trout,
it can be estimated with ICE. Raw acute data are
required to use ACE for chronic value estimations.
If these data are not available for the species in
question, raw data from an acute rainbow trout test
could be used in ACE, applying a factor of 0.5x to
the estimated chronic value to calculate chronic
toxicity to the endangered aquatic vertebrate.
Species sensitivity distributions (SSDs) conducted
on observed and estimated acute toxicity data
indicated that estimated data can be used to
accurately and precisely determine 5th percentile
values that include the most sensitive species
tested, particularly ICE-estimated data using the
best interspecies correlation with appropriate
surrogate species (i.e., in this case, fathead
minnow, rainbow trout, or sheepshead minnow).
Using fathead minnow as the only surrogate for
ICE estimates or QSAR estimated data, tended
to become less conservative and more variable
in determining 5th percentile values, and thus,
less likely to include the 96-h LC50 for the most
sensitive species tested. Dyer et al. (2006) also
found SSDs for ICE-estimated data corresponded
well to SSDs for observed data, but the results
were more variable as they used only one
surrogate species per SSD determination. Various
approaches in estimating chronic toxicity data
were also conducive to estimating 5th percentile
SSDs that corresponded well to SSDs for
observed data. Although small acute and chronic
data sets were used, the SSD method of Solomon
et al. (1996, 2000) was conservative and produced
the most consistent SSDs for including the most
sensitive species. It may be that the Solomon
methodology is best when using small data sets.
-------
The results of these studies suggest that
threatened and endangered species do not
appear to be universally more sensitive to
contaminant exposure than other species frorri a
toxicological standpoint. However, it is important
to consider the consequences of additional
pressure on species populations that are already
stressed. By definition, population numbers,
genetic variability, and habitat availability for
endangered species are low. Toxicant stress
that might be assimilated by a healthy population
could have a much more severe effect on a
fragile and sensitive population. The removal
of a portion of a robust population via fishing
pressure, habitat alteration, or even toxicant
stress has been deemed to be legal under
current regulations. The 1985 guidelines
(Stephan et al. 1985) state that because aquatic
ecosystems can tolerate some stress and
occasional adverse effects, protection of all
species at all times and places is not deemed
necessary.; If criteria are developed with that in
mind, then loss of individuals within threatened
and endangered species populations may
become very real. Considering that threatened
and endangered species, at least vertebrates,
do not appear to be significantly more sensitive
to chemicals than other species, the data and
modelling methodology presented herein can be
used to develop approaches for identifying critical
chemical concentrations for those as well as
other species having little or no data.
1.0-
0.8
-Q
JS 0.6
o
1 0.4
E
O
0.2-
0.0-
Carbaryl
• G, pseudolimnaeu^,
• D. magna —
Boreal toad
Bullfrog
Pickerel frog
'Plains leopard frog
Oregon spotted frog
/ W Green frog
/Spotted salamander
> Southern leopard frog
' Desert pupfish
1 Bluegill
' Gray tree frog
i Fathead minnow
' Foothill yellow-legged frog
1 Leon Springs pupfish
1 Razorback sucker
»Cape Fear shiner
1 Spotfin chub
Bonytail chub
1 Colorado pikeminnow
'Sheepshead minnow
1 Lahontan cutthroat trout
1 Greenthroat darter
' Shortnose sturgeon
f O Rainbow trout
1 Fountain darter
• Greenback cutthrout trout
* Apache trout
0.001
""I
0.01
11 Mill
0.1
10
' I
100
48-h EC50 or 96-h LC50 (mg/L)
Figure 1a. Species sensitivity (static acute toxicity) distribution for carbaryl (o = designated
surrogate test species for comparison). Dashed line = lower 95% confidence limit. D. magna
and G. pseudolimnaeaus (Mayer and Ellersieck 1986) were added as reference points.
-------
o
<£
1.0 -
0.8 -
0.6-
0.4 -
0.2 -
0.0 -
Copper
•Bluegill
Fathead minnow
• Colorado pikeminnow
> Greenthroat darter
/ / • Razorback sucker
/ /• Bonytail chub
/ */Gila topminnow
Southern leopard frog
Spotfin chub
•/Cape Fear shiner
'Apache trout
Shortnose sturgeon
Boreal toad
G. pseudolimnaeus
Lahontan cutthroat trout
D. magna
O Rainbow trout
• Fountain darter
\
1.0
10 100
48-h EC50 or 96-h LC50 (ug/L)
1,000
10,000
Figure 1b. Species sensitivity (static acute toxicity) distribution for copper (o = designated
surrogate test species for comparison). Dashed line = lower 95% confidence limit. D, magna
and G. pseudolimnaeus (USEPA 1985) were added as reference points.
1.0 -
0.8 -
I °-6
o
0.4-
0.2 -
0.0 -
4-Nonylphenol
Southern leopard frog
' Sheepshead minnow
Fathead minnow
Bonytail chub
Gila topminnow
Colorado pikeminnow
Cape Fear shiner
Bluegill
Rainbow trout
Razorback sucker
Greenthroat darter
Lahontan cuttroat trout
Apache trout
Greenback cutthroat trout
Boreal toad
Leon Springs pupfish
D. magna
Spotfin chub
Fountain darter
Shortnose sturgeon
1.0
10 100
48-h EC50 or 96-h LC50 (ug/L)
r-r-q
1,000
10,000
Figure 1c. Species sensitivity (static acute toxicity) distribution for 4-nonylphenol (o = designated
surrogate test species for comparison). Dashed line = lower 95% confidence limit. D. magna
(U.S. EPA 2003) was added as a reference point.
10
-------
S
CL
1.0 -
0.8
0.6 H
0.4 -
0.2 -
0.0
Pentachlorophenol
O Sheepshead minnow
G. pseudolimnaeus
Leon Springs pupfish
• Boreal toad
. magna
Bluegill
Gila topminnow
Southern leopard frog
Razorback sucker
Fathead minnow
Spotfin chub
Bonytail chub
Cape Fear shiner
Greenback cutthroat trout
Lahontan cutthroat trout
Greenthroat darter
Rainbow trout
' Colorado pikeminnow
f Apache trout
Fountain darter
'Shortnose sturgeon
T~
1.0
10 100
48-h EC50 or 96-h LC50 (ug/L)
1,000
10,000
Figure 1d. Species sensitivity (static acute toxicity) distribution for pentachlorophenol (o - designated
surrogate test species for comparison). Dashed line = lower 95% confidence limits. D. magna and G.
pseudolimnaeus (USEPA 1986) were added as reference points.
1.0 -
0,8
I?
15
o
•I
JS
3
E
o
0.4-
0.2
o.o-
Permethrin
Bonytail chub
'Colorado pikeminnow
Sheepshead minnow
> Leon Springs pupfish
Southern leopard frog
> Giia topminnow
fO Fathead minnow
' Razorback sucker
Bluegill
/• cApe Fear shiner
I •/Fountain darter
'Rainbow trout
'Greenthrout darter
Greenback cutthroat trout
' Apache trout
[ Lahontan cutthroat trout
' Spotfin chub
•prmagna
Jolimnaeus
I—
0.01
0.1
1.0 10
48-h EC50 or 96-h LC50 (ug/L)
100
Figure 1e. Species sensitivity (static acute toxicity) distribution for permethrin (o = designated
surrogate test species for comparison). Dashed line = lower 95% confidence limit. D. magna
and G. pseudolimnaeus (Mayer and Ellersieck 1986) were added as reference points.
11
-------
1-0 -j Copper
0.8-
I 0.6
o
oi
£
3
O
0.4-
0.2 -
o.o-
.
'
/
•
X
• White perch
-_ •American eel
• Bluegill
Banded killfish
• Mozambique tilapia
• Pumpkinseed
, Striped shiner
= . Sockeye salmon
/(.yOrangetnroat darter
•/Atlantic salmon
/J^Mosquitofish
Striped bass
Johnny darter
/ W Common carp
' •Goldfish
Chiselmouth
Guppy
Fathead minnow
-. Fantail darter
/ 9 Brook trout
OGreenthrout darter
/ IrBlacknose dace
. O Rainbow darter
/ W Creek chub
. f Central stoneroller
/ g Bluntnose minnow
Brown bullhead
Coho salmon
Cutthroat trout
Rainbow trout
Chinook salmon
'Scud (G. Dulex)
Daphnid (D. pulex)
'Scud (G, pseudolimnaeus)
Daphnid/D. magna)
Daphnid (C. reticulate)
Fountain darter
Northern squawfish
Daphnid (D. pulicaria)
I
0.1
1.0
10 100
48-h EC50 or 96-h LC50 (ug/L)
1000
10,000
Figure 2. Sensitivity of darters (o) relative to other aquatic species (• ) for copper (data from U.S. EPA
1985). Dashed line = lower 95% confidence limit.
12
-------
o
QL
-------
1.0 -
0.8 -
£ 0.6 -
o
£
I 0.4-
o
0,2 -
0.0 -
Permethrin B
Observed Data
Bonytail chub/^t0sheepshead minnow
Colorado pikeminnow/ ^Leon Springs pupfish
Sheepshead minnow//«OBonytail chub
Leon Springs pupfistr • ©Colorado pikeminnow
Southern leopard frog^J OSouthern leopard frog
Gila topminnowT»Gila topminnow
Fathead minnow/•Fathead minnow
Razorback suckerQ/Razorback sucker
Cape Fear shinerQ •//Greenback cutthroat trout
Fountain darterO »/Greenthroat darter
Rainbow troutO 9 /Cape Fear shiner
Greethroat darterQ f /Rainbow trout
Greenback cutthroat trout 9/ /Fountain darter
Apache troutQ^ii/Apache trout
Lahontan cutthroat troutO" /•Spotfin chub
Spotfin chub ,*• " j*3 • Lahontan cutthroat trout
Estimated Data
Surrogate
• Fathead minnow
I
0.1
1.0
10
96-h LC50 (ug/L)
100
1 ' i
1000
Figure 3b. Species sensitivity distributions for observed acute toxicity data and ICE-based
estimated data using fathead minnow as the only surrogate species. Species common names to
left of curve are observed data (o—) and species names to right of curve are estimated data <• —);
when only one value exists between two species ( •), the values are approximately the same.
14
-------
1.0-
0.8-
°-6
2
a.
3
o
0.4
0.2-
0.0
Permethrin C
Observed Data
Bonytail chub/'pOSheepshead minnow
Xxf
Colorado pikeminnow/^OLeon Springs pupfish
Sheepshead minnow^ • QBonytail chub
Leon Springs pupfisbMl OColorado pikeminnow
Southern leopard frog/fr O Gila topminnow
Gila topminnowAO Southern leopard frog
Fathead minnow I'9Q Fathead minnow
Razorback sucker
-------
UJ
I
S
^
o
100
90 -
80
3 70
60 -
50-
i
A
i
B
40
Daphnids
Rainbow
Trout
Gammarids
SPECIES
Figure 4. Test species required to include most sensitive species for acute toxicity tests (A-E represents
randomly selected other species). Cumulative % = species combinations including the most sensitive
species x% of the time, beginning with daphnids only (e.g., testing only daphnids,rainbow trout and
gammarids can include the most sensitive species 87.5% of the time).
Temperature
pH
Fish size-extreme
Fish size - normal (ASTM)
Life stage • invertebrates
Life stage - fish
Source-fish
Diet-fish
Pesticide formulation
Static vs. flo-thru
Species within a chemical
Chemicals within a species
•10123456 9
Orders of Magnitude
Figure 5. Range of effects and geometric mean of variables on acute toxicity of chemicals to aquatic
organisms. Range = highest EC/LC50 value + lowest value within a particular variable test and chemical;
geometric mean ( • ) of ranges among all tests for that variable (data from Mayer and Ellersieck 1986).
16
-------
Table 1. Surrogate, endangered, and related species tested with carbaryl, copper,
4-nonylphenol, pentachlorophenol, and permethrin (Bridges et al. 2002, Dwyer et al.
1999a,b, 2000a, Sappington et al. 2001).
Family
Species
Family
Species
Acipenseridae
Shovelnose sturgeon**
Scaphirhynchus platorynchus
Shortnose sturgeon ***
Acipenser brevirostrum
Atlantic sturgeon ***
Acipenser oxyrhynchus
Cyprinidae
Fathead minnow*
Pimephales promelas
Bonytail chub ***
Gila elegans
Cape Fear shiner***
Notropis mekistocholas
Colorado pikeminnow ***
Ptychocheilus lucius
Spotfin chub ***
Cyprinella monacha
Castostomidae
Razorback sucker ***
Xyrauchen texanus
Salmonidae
Rainbow trout*
Oncorhynchus mykiss
Apache trout ***
Oncorhynchus apache
Lahontan cutthroat trout ***
Oncorhynchus clarki henshawi
Greenback cutthroat trout ***
Oncorhynchus clarki stomias
Cyprinodontidae
Sheepshead minnow*
Cyprinodon variegatus
Desert pupfish ***
Cyprinodon macularius
Leon Springs pupfish ***
Cyprinodon bovinus
Poecilidae
Gila topminnow ***
Poeciliopsis occidentalis
Percidae
Fountain darter ***
Etheostoma fonticola
Greenthroat darter***
Etheostoma lepidum
Bufonidae
Boreal toad ***
Bufo boreas
Hylidae
Gray tree frog*
Hyla versicolor
Ranidae
Plains leopard frog*
Rana blairi
Foothill yellow-legged frog*
Rana boyli
Bullfrog*
Rana catesbeiana
Green frog*
Rana clamitans
Pickerel frog*
Rana palustris
Oregon spotted frog,***
Rana pretiosa
Southern leopard frog**
Rana sphenocephala
Ambystomatidae
Spotted salamander
Ambystoma maculatum
* Surrogate test species for studies included.
** Species identified as surrogates in U.S. Fish and Wildlife Service Recovery Plans.
*** State and/or federally listed (threatened or endangered species).
*Non-listed species.
1-7
-------
Table 2. Acute static toxicity of carbaryl to surrogate test species and endangered species.1 Number
of tests = 1-6/species; replicates/test'= 2-3.
Family
Species common name
96-h LC50 (ua/LV
95% CU
Acipenseridae
Shortnose sturgeon **
Cyprinidae
Fathead minnow*
Bonytail chub **
Cape Fear shiner **
Colorado pikeminnow **
Spotfin chub **
Catostomidae
Razorback sucker **
Salmonidae
Rainbow trout*
Apache trout **
Greenback cutthroat trout **
Lahontan cutthroat trout **
Cyprinodontidae4
Sheepshead minnow*, 2ppt
Sheepshead minnow*, 15ppt
Desert pupfish**,15ppt
Leon Springs pupfish **, 2ppt
Leon Springs pupfish **, 15ppt
Poecillidae
Gila topminnow **
Percidae
Fountain darter **
Greenthroat darter**
Bufonidae
Boreal toad**
Hylidae
Gray tree frog
Ranidae
Plains leopard frog
Foothill yellow-legged frog
Bullfrog
Green frog
Pickerel frog
Oregon spotted frog **
Southern leopard frog
Ambystomatidae
Spotted salamander
19491 .i.j .k
5171bcds
3044° de '9
3038 e- ' 9 h '• '
34-jgd.e.f.g.h
1866hJ--;-k
1435k
1553''K
2221 9 h ' ' K
4362 cd el
2511 fg.h.i.j.
7714abc
454Qc.de.!
2017 hi'k
>3000
1615'ik
12303a
6214abcd
11477a
4592 cde.<
11830a
9523 "•"
11487a
9632 ab
7964abc
8034 abc
1560-2354
4922 - 5437
2723-3413
3551 -5197
2803 - 3296
3123-37373
4087 - 4626
1766- 1972
1292- 1594
1368- 1760
2076 - 2380
3839-5120
2175-2905
6082-12332
4043-5179
1696-2390
1234-2362
1732-2872
5757-6710
10621
3956-
10709
8617-
10710
8794-
6916-
- 12469
5166
- 13110
10553
- 123163
10586
9232
7294 - 8879
'Surrogate test species.
"Listed species.
1 Means with same letter are not significantly different (p > 0.05).
2 Confidence limits.
3 Confidence limits estimated by Spearman-Karber techniques (Hamilton et al. 1977 a,b).
4 Cyprinodontids were tested under two salinity conditions, 2 or 15 parts per thousand salinity (ppt).
18
-------
Table 3. Acute static toxicity of copper and 4-nonylphenol to surrogate test species and endangered species.1
Number of tests = 1-6/species; replicates/test = 2-3. Copper values adjusted to 50 mg/L hardness
(US EPA 1985) except for Cyprinodontidae.
Family
Species common name
Acipenseridae
Shovelnose sturgeon
Atlantic sturgeon**
Shortnose sturgeon**
Cyprinidae
Fathead minnow*
Bonytail chub**
Cape Fear shiner**
Colorado pikeminnow**
Spotfin chub**
Catostomidae
Razorback sucker**
Salmonidae
Rainbow trout*
Apache trout**
Greenback cutthroat trout**
Lahontan cutthroat trout**
Cyprinodontidae4
Sheepshead minnow*, 2ppt
Sheepshead minnow*, 15ppt
Leon Springs pupfish**, 2ppt
Leon Springs pupfish **, 15ppt
Poecillidae
Gila topminnow**
Percidae
Fountain darter**
Greenthroat darter**
Bufonidae
Boreal toad**
Ranidae
Southern leopard frog
Cop|
96-h LC50 fua/L
51 c'd'e
198
25 e, f. g
144"
6gb,c,d
35d.e.f.g
132 b
35d,.e,f,g
84 b< c' d
ige.f.g
>9.5
21 f,9
630 a
>204
1306a
>204
4g c. d, e, f
18 9
95 b c-
23 e-'-g
44 c. d, e, f. g
aer
J1 95% CL2
37
16
20
133
65
31
117
31
76
17
21
20
518
1018
42
15
66
21
39
-66
-21
-31
-158
-74
-40
-150
-393
-92
-20
-28
-23
-791
-2384
-56
-21
-226
-25
-49
4-Nonylphenol
96-h LC50 (ua/U1 95% CL2
81 'J
272 b' Ci • e
264 b> Ci di e' *
215c.d,e,f
231 c'd'e-f
98g,h,i,.j
182c,d,e,f,g
1 91 °! ^ e' *• ^
161 d,e,f,g,h
153 e'f'9'hj
•\ 63 c, d, e, f, g
472 a, b
30Qa,b.c,d
553 a
132f.g,h,i
243b.c,d,e,f
89 ^u
1 79 c, d, e, f, g
137f,g,h,i
332a,b,c
260 - 283
241 - 290
176-259
219-2443
93-1033
170- 193
183-200
148- 175
141 -169
149-178
422 - 5283
259 - 344
417- 1230
91 - 167
74-109
133-291
131 - 1443
289 - 392
* Surrogate test species.
**Listed species.
1 Means with same letter are not significantly different (p > 0.05).
2 Confidence limits.
3 Confidence limits estimated by Spearman-Karber techniques (Hamilton et al. 1977 a,b).
4 Cyprinodontids were tested under two salinity conditions — 2 or 15 parts per thousand salinity (ppt).
19
-------
Table 4. Acute static toxicity of pentachlorophenol and permethrin to surrogate test species
and endangered species. 1Number of tests = 1 -6/species; replicates/test = 2-3.
Pentachlorophenol values adjusted to pH 6.5 (U.S EPA 1986) except for Cyprinodontidae.
Family
Species common name
Acipenseridae
Shortnose sturgeon**
Cyprinidae
Fathead minnow*
Bonytail chub**
Cape Fear shiner**
Colorado pikeminnow**
Spotfin chub**
Catostomidae
Razorback sucker**
Salmonidae
Rainbow trout*
Apache trout**
Greenback cutthroat trout**
Lahontan cutthroat trout**
Cyprinodontidae5
Sheepshead minnow*, 2ppt
Sheepshead minnow*. 15ppt
Leon Springs pupfish**,2ppt
Leon Springs pupfish**,15ppt
Poecillidae
Gila topminnow**
Percidae
Fountain darter**
Greenthroat darter**
Bufonidae
Boreal toad**
Ranidae
Southern leopard frog
Pentachlorophenol
96-h LC50 (ua/L)1 95%
11"
40c.d.e
37c.d.e.f
35def
OH s f .g. h
39 c. d e
4Qc.d.e
25 d- e. f. g
17fg.h
33d.e.f..g
28 d e '- s
40 c .d.e
191 a
op b. c
107 ab
56D.C.d
15g.h
26de'9
98 ab
4gt.c.d
9.3-
38-
35-
32-
20-
37-
39-
24-
15-
17-
26-
163-
73-
82-
49-
12-
24-
86-
44-
CL2-
• 14
43
393
383
23
413
413
26
18
181
30
224
92
125
65
18
293
113
56
Permethrin
96-h LC50 (ua/LV1 95%CL2
NC4
9.5 cd
39 a
3.5 e-f
26 b.=
1.7'
6.1 d e
3.3 8f
2.0'
2.8 e-i
1.7'
17abc
23 a b
19 a°c
21 a.b
10b.c.d
3.3ef
2.8 ef
> 10
20 a be
8.9
27
3.2
20
1.5
5.5
3.1
1.8
1.4
1.5
15
16
2.8
2.5
16
-10
-80
-3.73
-42
- 1.9
-6.8
-3.5
-2.2
-4.2
- 1.8
- 193
-22
-4.0
-3.2
-26
* Surrogate test species.
"Listed species.
1 Means with same letter are not significantly different (p > 0.05).
2 Confidence limits.
3 Confidence limits estimated by Spearman-Karber techniques (Hamilton et al. 1977 a,b).
4 Could not be calculated.
5 Cyprinodontids were tested under two salinity conditions, 2 or 15 parts per thousand salinity (ppt).
20
-------
Tables. Acute toxicity sensitivity rankings by 96-h LC50. Rank = sum of
rankings among chemicals within a species/n for chemicals
having a 96-h LC50.
Species common name 96-h LC50 rank1
Atlantic sturgeon* 2.02
Fountain darter* 2.4
Shortnose sturgeon* 3.0
Apache trout* 3.6
Greenback cutthroat trout* . 4.8
Rainbow trout 5.2
Lahontan cutthroat trout* 5.4
Spotfin chub* 6.6
Greenthroat darter* 7.4
Shovelnose sturgeon 9.02
Cape Fear shiner* 9.0
Leon Springs pupfish*,15ppt 9.8
Razorback sucker* 10.6
Colorado pikeminnow* 10.8
Gila topminnow* 11.0
Boreal toad* 11.2
Bonytail chub* 12.2
Fathead minnow 13.2
Southern leopard frog 13.6
Sheepshead minnow, 2ppt 14.0
Sheepshead minnow, 15ppt 14.2
Leon Springs pupfish*, 2ppt 15.4
Desert pupfish* 18.03
Other amphibians >18.03
*Listed species.
10nly three values were significantly (p < 0.05) less than that for rainbow
trout (4-nonylphenol, shovelnose sturgeon and fountain darter, pentachloro-
phenol, shortnose sturgeon).
2 One test with copper.
3 One test with carbaryl.
21
-------
Table 6. Chronic toxicity (ug/L) of copper (50 mg/L hardness) and
pentachlorophenol (6.5 pH) to fathead minnow, spotfin
chub, rainbow trout, and fountain darter.1 Numbers in
parentheses are 95% confidence limits.
Species common
name
Fathead minnow
Spotfin chub
Rainbow trout
Fountain darter
Fathead minnow
Spotfin chub
Rainbow trout
Fountain darter
Tests Chronic Value
(n) Survival
Copper
3 9.5
(7.0-13)
1 11
•(8.1-15)
2 5.6
(4.2-7.7)
2 2.7
(2.1-3.5)
Pentachlorophenol
4 27a
(20-38)
1 33a
(24-45)
2 17°
(12-24)
1 8.7b
(6.4-12)
(NOEC-LOEC)2
Growth3
2.8
(2.0-3.9)
5.6
(3.9-8.1)
4.2
(3.0-5.6)
2.7
(2.1-3.5)
27a
(20-38)
8.0°
(5.7-11)
8.3°
(5.9-12)
3.9°
(2.5-6.4)
1 Means with same letter are not significantly different (p <0.05).
2 Geometric mean of chronic values (NOEC, No-observed-effect
concentration; LOEC, lowest observed effect concentration).
3 Dry weight.
22
-------
Table 7, Interspecies correlations (X2 = a + bXJ for all species and five chemicals
with fathead minnow as the surrogate species. X1 = log 96-h LC50 for
fathead minnow and X2 = log 96-h LC50 for other species in |jg/L
Family
Species common name
Acipenseridae
Shortnose sturgeon
Cyprinidae
Bonytail chub
Cape Fear shiner
Colorado pikeminnow
Spotfin chub
Catostomidae
Razorback sucker
Salmondiae
Apache trout
Greenback cutthroat trout
Lahontan cutthroat trout
Rainbow trout
Cyprinodontidae
Leon Springs pupfish
Sheepshead minnow
Poecillidae
Gila topminnow
Percidae
Fountain darter
Greenthroat darter
Bufonidae
Boreal toad
Ranidae
Southern leopard frog
n
4
5
5
5
5
5
5
4
5
5
4
4
4
5
5
4
5
Intercept (a)
-0.791
0.560
-0.471
0.351
-0.684
-0.175
-0.596
-0.259
-0.676
-0.414
0.724
0.888
0.284
-0.508
-0.349
-0.496
0.091
Slope (b)
1 .092**
0.745**
1.098**
0.823**
1.129**
1.017**
1 .033**
0.959*
1 .096**
0.993**
0.674*
0.684*
0.793
0.979**
1 .026**
1.166
0.973**
r
0.993
0.947
0.980
0.967
0.973
0.996
0.980
0.983
0.968
0.962
0.970
0.968
0.907
0.982
0.989
0.882
0.952
*Slope significant from zero, p < 0.05.
**Slope significant from zero, p < 0.01.
23
-------
Table 8. Interspecies correlations (X2 = a + bX,) for all species and five chemicals
with rainbow trout as the surrogate species. X, = log 96-h LC50 for
rainbow trout and X2 = log 96-h LC50 for other species in ug/L.
Family
Species common name
Acipenseridae
Shortnose sturgeon
Cyprinidae
Bonytail chub
Cape Fear shiner
Colorado pikeminnow
Fathead minnow
Spotfin chub
Catostomidae
Razorback sucker
Salmondiae
Apache trout
Greenback cutthroat trout
Lahontan cutthroat trout
Cyprinodontidae
Leon Springs pupfish
Sheepshead minnow
Poecillidae
Gila topminnow
Percidae
Fountain darter
Greenthroat darter
Bufonidae
Boreal toad
Ranidae
Southern leopard frog
n
4
5
5
5
5
5
5
5
4
5
4
4
4
5
5
4
5
Intercept (a)
-0.184
0.925
0.033
0.833
0.550
-0.133
0.360
-0.118
0.019
-0.195
0.952
1.092
0.644
-0.049
0.211
-0.054
0.520
Slope (b)
1.022*
0.719*
1.080**
0.750*
0.932**
1.093**
0.962**
1.014**
0.973**
1.090**
0.665*
0.690*
0.778**
0.958**
0.959**
1.192
0.968**
r
0.967
0.944
0.995
0.910
0.962
0.972
0.973
0.993
0.996
0.994
0.956
0.976
0.997
0.991
0.954
0.938
0.976
*Slope significant from zero, p < 0.05.
"Slope significant from zero, p < 0.01.
24
-------
Table 9. Interspecies correlations (X2 = a + bXJ for all species and five chemjcals
with sheepshead minnow as the surrogate species. X, = log 96-h LC50
for sheepshead minnow (15ppt salinity) and X2 = log 96-h LC50 for other
species in |jg/L.
Family
Species common name
Acipenseridae
Shortnose sturgeon
Cyprinidae
Bonytail chub
Cape Fear shiner
Colorado pikeminnow
Fathead minnow
Spotfin chub
Catostomidae
Razorback sucker
Salmondiae
Apache trout
Greenback cutthroat trout
Lahontan cutthroat trout
Rainbow trout
Cyprinodontidae
Leon Springs pupfish
Poecillidae
Gila topminnow
Percidae
Fountain darter
Greenthroat darter
Bufonidae
Boreal toad
Ranidae
Southern leopard frog
n
3
4
4
4
4
4
4
4
4
4
4
4
3
4
4
3
4
. Intercept (a)
-2.911
-0.017
-1.646
-0.360
-1.081
-2.050
-1.304
-1.705
-1.417
-1.906
-1.419
-0.110
-0.539
-1.489
-1.569
-2.563
-0.760
slope(b)
1.838
0.959
1 .537**
1.054
1.371*
1.631**
1.417*
1.439*
1 .366**
1 .559**
1 .380*
0.968**
1.105
1 .345*
1 .442*
1 .950
1.308*
r
0.975
0.887
0.986
0.875
0.968
0.999
0.982
0.973
0.989
0.989
0.976
0.985
0.952
0.973
0.983
0.995
0.948
*Slope significant from zero, p < 0.05.
**Slope significant from zero, p < 0.01.
25
-------
Species common mime. Species number 1
10 11
13 14 15 Id 17 IS
Colorado piU: mimi
0.97 V1 0.97-1 '' O.l'-'l
0,99s1' l.OOO1'
SlKxpsln.-;nl iniiiiiuu 1 Sppl
l7
•i •!
-------
Table 11. Observed and estimated 96-h LCSOs (ug/L) for five chemicals and 15 aquatic vertebrates using
interspecies correlation analysis (ICE, Asfaw et al. 2003) and surrogate species (fathead minnow,
rainbow trout, or sheepshead minnow) acute toxicity values. The surrogate species model having
the highest r value was used for estimations.
Estimated species
Shortnose sturgeon
Bonytail chub
Cape Fear shiner
Colorado pikeminnow
Spotfin chub
Razorback sucker
Apache trout
Greenback cutthroat
trout
Lahontan cutthroat
trout
Leon Springs pupfish
Gila topminnow
Fountain darter
Greenthroat darter
Boreal toad
Southern leopard frog
Surrogate
Species
FHM1
FHM
RBT2
FHM
SHM3
FHM
RBT
RBT
RBT
SHM
RBT
RBT
FHM
SHM
RBT
Carbaryl
19494
(1837)5
3044
(2121)
4264
(3678)
3038
(2554)
3416
(3127)
4325
(3997)
1435
(1580)
1553
(1591)
2221
(2346)
2017
(1517)
>3000
1615
(1215)
2143
(2892)
12303
(11660)
7964
(4856)
Copper
25
(37)
69
(147)
35
(26)
132
(134)
35
(64)6
84
(105)
25
(15)
>9.5
21
(16)
>204
48
(44)
18
(15)
95
(73)
23
(197)
44
(57)
4-Nonylphenol
81
(74)
264
(236)
215
(314)
231
(226)
98
(98)
182
(200)
161
(157)
153
(173)
163
(196)
132
(194)
243
(262)
89
(137)
179
(141)
137
(185)
332
(535)
Pentachloro-
phenol
11
(9.1)
37
(57)
35
(35)
21
(47)
39
(47)
40
(28)
17
(20)
33
(24)
28
(21)
107
(125)
56
(54)
15
(20)
26
(20)
98
(77)
49
(75)
Permethrin
ND8
39
(19)
3.5
(3.9)
26
(14)
1.7
(1.5)
6.1
(6.6)
2.0
(2.6)
2.8
(3.3)
1.7
(2.3)
21
(16)
10
(11)
3.3
(2.8)
2.8
(4.5)
>10
20
(11)
1 FHM = fathead minnow.
2 RBT = rainbow trout.
3 SHM -sheepshead minnow.
4 Observed 96-h LC50.
5 Estimated 96-h LC50 using ICE (Asfaw et al. 2003).
6 Copper value not available for sheepshead minnow, fathead minnow model and copper value were used.
7 Copper value not available for sheepshead minnow, rainbow trout model and copper value were used.
27
-------
Table 12. Observed and estimated (ACE, Ellersieck et al. 2003)
chronic toxicities (ug/L) of copper (50 mg/L hardness) and
pentachlorophenol (6.5 pH) for fathead minnow, spotfin
chub, rainbow trout, and fountain darter. Predicted no-effect
chronic values were determined using raw acute toxicity
data and the software ACE (Ellersieck et al. 2003).1
Chronic Value (NOEC-LOEC)2 ACE (95% CU
Species
Fathead minnow
Fathead minnow4
Spotfin chub
Rainbow trout
Rainbow trout5
Fountain darter
Fathead minnow
Spotfin chub
Rainbow trout
Fountain darter
Survival
9.5
(7.0-13)
6.9
(5.4-8.9)
11
(8.1 - 15)
5.6
(4.2 - 7.7)
2.7
(2.1 -3.5)
27a
(20 - 38)
33a
(24 - 45)
17b
(12-24)
8.7b
(6.4- 12)
Growth3
Copper
2.8
(2.0 - 3.9)
6.9
(5.4 - 8.9)
5.6
(3.9-8.1)
4.2
(3.0 - 5.6)
2.7
(2.1 -3.5)
Pentachlorophenol
27a
(20 - 38)
8.0b
(5.7-11)
8.3b
(5.9- 12)
3.9b
(2.5 - 6.4)
3.5
(0.9 - 8.2)
11
(2.8- 19)
6.2
(2.5-9.9)
2.3
(-50 - 55)
1.4
(0.5 - 4.5)
1.6
(0.4 - 2.8)
10
(4.1 -16)
11
(4.9- 18)
13
(3.4 - 22)
4.1
(1.2-6.9)
1 Means with same letter are not significantly different (p < 0.05). ACE
estimates were not analyzed since each is a single value.
2 Geometric mean of chronic values (NOEC, no-observed effect
concentration ; LOEC, lowest observed effect concentration).
3 Dry weight.
4 Data from Spehar and Fiandt (1986).
5 Data from Stubblefield (1990).
28
-------
Table 13. Calculated 5lh percentile values for observed and estimated acute toxicity data (96-h LCSOs
in ug/L) using three methods (Solomon et al. 1996, 2000; Aldenberg and Jaworska 2000;
Stephanetal. 1985).
Chemical
Observed
Data
ICE-FHM,
RBT, SHM1
ICE-
FHM2
Solomon etal. (1996,
Carbaryl
Copper
4-Nonylphenol
Pentachlorophenol
Permethrin
Carbaryl
Copper
4-Nonylphenol
Pentachlorophenol
Permethrin
1073
13
90
11
1.1
1809
24
133
22
2.7
993
12
83
12
1.1
Aldenberg
1655
24
134
24
2.4
1252
35
73
8.0
1.5
ECOSAR-
FHM35
2000)
3721
-
39
128
1.4
ASTER-
FHM45
-
-
24
45
-
and Jaworska (2000)
1830
NC7
114
14
3.1
6019
-
NC
189
2.9
-
-
NC
NC
-
Stephanetal. (1985)
Carbaryl
Copper
4-Nonylphenol
Pentachlorophenol
Permethrin
1409
18
81
12
1.6
1250
15
77
11
1.3
1372
37
75
9.4
2.0
4207
-
41
128
1.9
-
-
26
47
-
29
-------
Table 14. Calculated 5:ilpercentile values (ug/L) for observed and estimated chronic toxicity data using
three methods (Solomon et al. 1996, 2000; Aldenberg and Jaworska 2000; Stephan et al.
1985).
Chemical
Copper
Pentachlorophenol
Observed Observed Data Observed Data Observed Data
Data1 +ACE FD, SFC2 +ICE/ACR FD, FHM, Dm +ICE/
SFC3 AC Pi4
Solomon et al.
2.3 1.9
2.5 2.8
(1996.2000)
2.3
2.8
1.2
3,2
Aldenberg and Jaworska (2000)
Copper
Pentachlorophenol
Copper
Pentachlorophenol
6.8 6.5
6.9 7.7
Stephan et
2.3 1.4
0.16 0.75
6.8
6.5
al. (1985)
2.4
0.59
3.2
7.9
0.45
1.2
1 Chronic data from criteria document (U.S. EPA 1985, 1986) plus chronic data for fountain darter (FD)
and spotfin chub (SFC).
2 Chronic data from criteria document plus chronic data for fountain darter and spotfin chub estimated
-by ACE (Ellersieck et al. 2003).
3 Chronic data from criteria document plus chronic data for fountain darter and spotfin chub estimated
by ICE for acute values followed with division by ACR for estimated chronic values.
4 Chronic data for Daphina magna and fathead minnow from criteria document with acute values for all
other species estimated by ICE followed with division by ACR for estimated chronic values.
30
-------
REFERENCES
Aldenberg.T. and J. Jaworska. 2000. Uncertainty
of the hazardous concentration and fraction
affected for normal species sensitivity distributions.
Ecotoxicol. Environ. Saf. 46:1-18.
American Society for Testing and Materials.
2003a. Standard guide for conducting acute
toxicity tests on test materials with fishes,
macroinvertebrates, and amphibians (E729-96).
In: Annual book of ASTM standards. Vol. 11.05.
American Society for Testing and Materials, West
Conshohocken, PA.
American Society for Testing and Materials.
2003b. Standard guide for conducting early life-
stage toxicity tests with fishes (E1241 -98). In:
Annual book of ASTM standards. Vol. 11.05.
American Society for Testing and Materials, West
Conshohocken, PA.
Asfaw, A., M.R. Ellersieck, and F.L. Mayer. 2003.
Interspecies correlation estimations (ICE) for
acute toxicity to aquatic organisms and wildlife. II.
User manual and software. U.S. Environmental
Protection Agency Report No. EPA/600/R-03/106,
Washington, DC.
Augspurger, T, A.E. Keller, M.C. Black, W.G. Cope,
and F.J. Dwyer. 2003. Water quality guidance
for protection of freshwater mussels (Unionidae)
from ammonia exposure. Environ. Toxicol. Chem.
22:2569-2575.
Besser, J.M., F.J. Dwyer, C.G. Ingersoll, and N.
Wang. 2001. Early life-stage toxicity of copper
to endangered and surrogate fish species. U.S
Environmental Protection Agency Report No.
EPA/600/R-01/051, Washington, DC..
Besser, J.M., N.Wang, F.J. Dwyer, F.L. Mayer,
and C.G. Ingersoll. 2005. Assessing contaminant
sensitivity of endangered and threatened aquatic
species: Part II. Chronic toxicity of copper and
pentachlorophenol to two endangered species and
two surrogate species. Arch. Environ. Contam.
Toxicol. 48:155-165
Bridges, C.M., F.J. Dwyer, O.K. Hardesty, and D.W.
Whites. 2002. Comparative contaminant toxicity:
Are amphibian larvae more sensitive than fish?
Environ. Contam. Toxicol. 69:562-569
DeGraeve, G.M., J.D. Cooney, D.O. Mclntyre, T.L.
Pollock, N.G. Reichenbach, J.H. Dean, and M.D.
Marcus. 1991. Variability in the performance of the
seven-day fathead minnow (Pimephales promelas)
larval survival and growth test: An intra- and inter-
laboratory study. Environ. Toxicol. Chem. 10:1189-
1204.
Dwyer, F.J., F.L. Mayer, L.C. Sappington, D.R.
Buckler, C.M. Bridges, I.E. Greer, O.K. Hardesty,
C.E. Henke, C.G. Ingersoll, J.L Kunz, D.W. Whites,
T. Augspurger, D.R. Mount, and K. Hattala. 2005a.
Assessing contaminant sensitivity of endangered •
and threatened aquatic species: Part I. Acute toxicity
of five chemicals. Arch. Environ. Contam. Toxicol.
48:143-154.
Dwyer, F.J., O.K. Hardesty, C.E. Henke, C.G.
Ingersoll, D.W. Whites, T. Augspurger, T.J. Canfield,
D.R. Mount, and F.L. Mayer. 2005b. Assessing
contaminant sensitivity of endangered and
threatened aquatic species; Part III. Effluent toxicity
tests. Arch. Environ. Contam. Toxicol. 48:174-183.
Dwyer, F.J., O.K. Hardesty, C.G. Ingersoll, J.L. Kunz,
and D.W. Whites. 2000. Assessing contaminant
sensitivity of American shad, Atlantic sturgeon,
and shortnose sturgeon. Report to New York
Department of Environmental Conservation. U.S.
Geological Survey, Columbia Environmental
Research Center, Columbia, MO, 30 p.
Dwyer, F.J., O.K. Hardesty, C.E. Henke, C.G. Ingersoll,
D.W. Whites, and D.R. Mount. 1999a. Assessing
contaminant sensitivity of endangered and threatened
species: Toxicant classes. U.S. Environmental
Protection Agency Report No. EPA/600/R-99/098,
Washington, DC.
Dwyer, F.J., O.K. Hardesty, C.G. Ingersoll, and D.W.
Whites. 1999b. Assessing contaminant sensitivity of
Cape Fear shiner and spotfin chub. Interim Report to
U.S. Fish and Wildlife Service. Ecological Services,
Raleigh, NC, 20 p.
Dwyer, F.J., L.C. Sappington, D.R. Buckler, and S.B.
Jones. 1995. Use of surrogate species in assessing
contaminant risk to endangered and threatened
species. U.S. Environmental Protection Agency
Report No. EPA 600/R-96/029, Washington, DC.
31
-------
Dyer, S.D., D.J. Versteeg, S.E. Belanger, J.G.
Chaney, and F.L. Mayer. 2006. Interspecies
correlation estimates predict protective
environmental concentrations. Environ. Sci.
Technol. 40:3102-3111.
Ellersieck, M.R., A. Asfaw, F.L. Mayer, G.F.
Krause, K.Sun, and G. Lee. 2003. Acute-
to-chronic estimation (ACE v. 2.0) with time-
concentration-effect models: User manual and
software. U.S. Environmental Protection Agency
Report No. EPA/600/R-03/107, Washington, DC.
Fairchild, J.F., A.L. Allert, L.C. Sappington, and
B. Waddell. 2005. Chronic toxicity of un-ionized
ammonia to early life-stages of endangered
Colorado pikeminnow (Ptychocheilus lucius) and
razorback sucker (Xyrauchen texanus) compared
to the surrogate fathead minnow (Pimephales
promelas). Arch. Environ. Contam.Toxicol.
49:378-384.
Finney, D.J. 1978. Statistical methods in
biological assay. Griffin, London.
Hamilton, M.A., R.C, Russo, and R.V.
Thurston. 1977a. Trimmed Spearman
Karber method for estimating median lethal
concentrations in toxicity bioassays. Environ. Sci.
Technol.11:714-719.
Hamilton, M.A., R.C. Russo, and R.V. Thurston.
1977b. Correction. Environ. Sci. Technol.
12:417.
Hansen, J.A., J. Lipton, and P.G.Welsh. 2002.
Relative sensitivity of bull trout (Salvelinus
confluentus) and rainbow trout (Oncorhynchus
mykiss) to acute copper toxicity. Environ. Toxicol.
Chem. 21:633-639.
Lemke, A.E. 1981. Inter-laboratory comparison
-Acute testing set. U.S. Environmental
Protection Agency Report No. EPA/600/3-81-
005, Duluth, MN.
Mayer, F.L. and M.R. Ellersieck. 1986. Manual
of acute toxicity: Interpretation and database
for 410 chemicals and 66 species of freshwater
animals. Resource Publication 160, U.S. Fish
and Wildlife Service, Washington, DC.
Mayer, F.L., K.Sun, G. Lee, M.R. Ellersieck,
and G.F. Krause. 1999. User guide: Acute-to-
chronic estimation (ACE). EPA/600/R-98/152. U.S.
Environmental Protection Agency, Washington, DC (+
software).
Mayer, F.L., M.R. Ellersieck, G.F. Krause, K. Sun, G.
Lee, and D.R. Buckler. 2002. Time-concentration-
effect models in predicting chronic toxicity from acute
toxicity test data. Pages 39-67, in M. Crane, M.C.
Newman, P.F. Chapman, and J. Fenlon, eds. Risk
Assessment with Time to Event Models. Lewis Publ.,
Boca Raton, FL.
Mummert, A.K., R.J. Neves, T.J. Newcomb, and D.S.
Cherry. 2003. Sensitivity of juvenile freshwater
mussels (Lampsilis fasciola, Villosa iris) to total
and un-ionized ammonia. Environ. Toxicol. Chem.
22:2545-2553.
Posthuma, L, G.W. Suter II, andT.PTraas. 2002.
Species sensitivity distributions in ecotoxicology.
Lewis Pub., Boca Raton, FL 587 p.
Russom, C.L., E.B. Anderson, B.E. Greenwood, and
A. Pilli. 1991. ASTER: An integration of the AQUIRE
data base and the QSAR system for use in ecological
risk assessments. Sci. Total Environ. 109/110:667-
670.
Russom, C.L., S.P. Bradbury, S.J. Broderius, D.E.
Hammermeister, and R.A. Drummond. 1997.
Predicting modes of toxic action from chemical
structure: Acute toxicity in the fathead minnow
(Pimephales promelas). Environ. Toxicol. Chem.
16:948-967.
Sappington, L.C., F.L. Mayer, FJ. Dwyer, D.R. Buckler,
J.R. Jones, and M.R. Ellersieck. 2001. Contaminant
sensitivity of threatened and endangered fishes
compared to standard surrogate species. Environ.
Toxicol. Chem. 20:2869-2876.
SAS Institute Inc. 2004. SAS/STAT 9.1 User's Guide.
SAS Institute Inc., Gary, NC.
Schimmel, S.C. 1981. Results: Inter-laboratory
comparison - Acute toxicity tests using estuarine
animals. U.S. Environmental Protection Agency
Report No. EPA/600/4-81-003, Gulf Breeze, FL.
Snedecor, G.W. and W.G. Cochran. 1989. Statistical
methods, 8th ed. Iowa State University Press, Ames,
IA.
32
-------
Solomon, K.R., D.B. Baker, R.P. Richards, K.R.
Dixon, S.J. Klaine, T.W. LaPoint, R.J. Kendall,
C.P. Weisskipf, J.M. Giddings, J.P. Giesy, L.W.
Hall, Jr., and W.M.Williams. 1996. Ecological
risk assessment of atrazine in North American
surface waters. Environ. Toxicol. Chem. 15:31-
76.
Solomon, K., J. Giesy, and P. Jones. 2000.
Probabilistic risk assessment of agrochemicals
in the environment. Crop Protection 19:649-655.
Spehar, R.L. and J.T. Fiandt. 1986. Acute
and chronic effects of water quality criteria-
based metal mixtures on three aquatic species.
Environ. Toxicol. Chem. 5:917-931.
Stephan, C.E., D.I. Mount, D.J. Hansen, J.H.
Gentile, G.A. Chapman, and W.A. Brungs. 1985.
Guidelines for deriving numerical national water
quality criteria for the protection of aquatic
organisms and their uses. (PB85-227049). U.S.
Environmental Protection Agency, Washington,
DC.
Stubblefield, W.A. 1990. An evaluation of the
acute toxicity of copper chloride (CuCI2) to brown
trout (Salmo trutta), rainbow trout (Oncorhynchus
mykiss), and mountain whitefish (Prosopium
Williamson!). Project 8505-088-901, ENSR
Consulting and Engineering, Ft. Collins, CO. 33
p. + appendices.
Urban, D.J. and N.J. Cook. 1986. Hazard
Evaluation Division standard evaluation
procedure: Ecological risk assessment. EPA-
540/9-85-001. Office of Pesticide Programs, U.S.
Environmental Protection Agency, Washington,
DC.
U.S. Environmental Protection Agency. 1978.
Water Quality criteria. Federal Register
43(97):21506-21518.
U.S. Environmental Protection Agency. 1985.
Ambient water quality criteria for copper-1984.
EPA 440/5-84-031. Office of Water Regulations
and Standards Division, Washington, DC.
U.S. Environmental Protection Agency.
1986. Ambient water quality criteria for
pentachlorophenol. EPA 440/5-86-009. Office
of Water Regulations and Standards Division,
Washington, DC.
U.S. Environmental Protection Agency. 1991.
Technical support document for water quality
based toxics control. U.S. Environmental
Protection Agency Report No. 505/2-90-001,
Washington, DC.
U.S. Environmental Protection Agency. 2001.
Environmental Effects Database (EEDB),
ECOTOX Database System. U.S. Environmental,
Protection Agency website, http://www.epa.gov/
ecotox/, Washington, DC.
U.S. Environmental Protedtion Agency. 2003.
Ambient aquatic life water quality criteria for
nonylphenol-Draft. EPA 882-R-03-029. Office of
Water, Washington, DC.
Wang, N., C.G. Ingersoll, O.K. Hardesty, C.D. Ivey,
J.L. Kunz, T.W. May, RJ. Dwyer, A.D. Roberts,
T. Augspurger, C.M. Kane, R.J. Neves, and
M.C. Barnhart. 2007. Acute toxicity of copper,
ammonia, and chlorine to glochidia and juveniles
of freshwater mussels (unionidae). Environ.
Toxicol. Chem. 26:2036-2047.
Wheeler, J.R. E.P.M. Grist, K.M.Y. Leung, D.
Morritt, and M. Crane. 2002. Species sensitivity
distributions; Data and model choice. Mar. Poll.
Bull. 45:192-202.
33
-------
v EPA
United States
Environmental Protection
Agency
Office of
Research and Development
Washington DC 20460
Official Business
Penalty for Private Use
$300
EPA/600/R-48/045
September 2008
Recycled/Recyclable
Printed \vith vegetable-based ink on
> paper that contains a minimum of
50% post consumer fiber content
processed chlorine free
------- |