United States Office of Water EPA 822-P-15-001
Environmental Protection 4304T July 2015
Agency
Draft
Aquatic Life Ambient Water Quality
^
^^
Criterion for ^^^
Selenium - Freshwater
2015 ^/
U.S. Environmental Protection Agency
Office of Water
Office of Science and Technology
Washington, D.C.
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TABLE OF CONTENTS
Table of Contents ii
List of Tables iv
List of Figures v
Notice vi
Foreword vii
Acknowledgements viii
Acronyms x
Executive Summary xi
1 Introduction and Background 1
1.1 History of the EPA Selenium AWQC for Aquatic Life 1
2 Problem Formulation 4
2.1 Overview of Selenium Sources and Occurrence 4
2.2 Environmental Fate and Transport of Selenium in the Aquatic Environment 9
2.2.1 Selenium Species in Aquatic Systems 9
2.2.2 Bioaccumulation of Selenium in Aquatic Systems 11
2.3 Mode of Action and Toxicity of Selenium 13
2.4 Narrow Margin between Sufficiency and Toxicity of Selenium 15
2.5 Interactions with Mercury 16
2.6 Assessment Endpoints 17
2.7 Measures of Effect 19
2.7.1 Fish Tissue 20
2.7.2 Water 22
2.7.3 Summary of Assessment Endpoints and Measures of Effect 23
2.7.4 Conceptual Model of Selenium Effects on Aquatic Life 25
2.7.5 Analysis Plan for Derivation of the Chronic Fish Tissue-Based Criterion Elements26
2.7.6 Analysis Plan for Derivation of the Fish Tissue Criterion Elements Duration 27
2.7.7 Analysis Plan for Derivation of Chronic Water-based Criterion Element 27
2.7.8 Analysis Plan for Derivation of the Water Criterion Elements Duration 30
2.7.9 Analysis Plan for Intermittent-Exposure Water-based Criterion Element Derivation
31
3 Effects Analysis for Freshwater Aquatic Organisms 32
3.1 Chronic Tissue-Based Selenium Criterion Element Concentration 32
3.1.1 Acceptable Studies of Fish Reproductive Effects of Four Most Sensitive Genera... 33
3.1.2 Summary of Acceptable Studies of Fish Reproductive Effects 39
3.1.3 Invertebrate Chronic Effects 42
3.1.4 Summary of Relevant Invertebrate Tests 45
3.1.5 Derivation of Tissue Criterion Element Concentrations 46
3.1.6 Selenium Fish Tissue Toxicity Data Fulfilling Minimum Data Needs 52
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3.2 Chronic Water Column-based Selenium Criterion Element 55
3.2.1 Translation from Fish Tissue Concentration to Water Column Concentration 55
3.2.2 Equation Parameters 63
3.2.3 Food-Web Models 75
3.2.4 Classifying Categories of Aquatic Systems 76
3.2.5 Deriving Protective Water Column Concentrations for Lentic and Lotic System
Categories 80
3.2.6 Derivation of Averaging Period for Chronic Water Criterion Element 85
3.3 Intermittent-Exposure Water Criterion Element: Derivation from the Chronic Water
Criterion Element 86
4 National Criterion for Selenium in Fresh Waters 90
4.1 Protection of Downstream Waters 94
5 Site-specific Criteria 95
6 Effects Characterization 97
6.1 Fish and Amphibians 97
6.1.1 Principles for Using Studies for which ECIQS Cannot Be Calculated 97
6.1.2 Acceptable Studies of Fish Reproductive Effects of Genera not the Four Most
Sensitive 98
6.1.3 Reproductive Effects in Catfish (Ictaluridae) 105
6.1.4 Reproductive Effects in Amphibians (Xenopus laevis) 110
6.1.5 Reproductive Studies Not Used in the Numeric Criterion Derivation 110
6.1.6 Salmo GMCV: EPA Re-analysis of a Key Study Used in Criterion Derivation 113
6.1.7 Impact of Number of Tested Species on Criterion Derivation 116
6.1.8 Comparisons between Concentrations in Different Tissues 116
6.1.9 Studies of Non-Reproductive Effects 117
6.1.10 Special conditions for consideration of primacy of water column criterion
elements over fish tissue criterion elements 117
6.1.11 Comparison of Fish Chronic Reproductive Effects and Chronic Non-Reproductive
Effects 122
6.2 Water 124
6.2.1 Validation of Translation Equation for Developing Water Column Concentrations
124
6.3 Protection of Threatened or Endangered Species 126
6.3.1 Special Consideration for Pacific Salmonid Juveniles 128
6.4 Aquatic-Dependent Wildlife is Beyond the Scope of this Aquatic Criteria Derivation
131
7 References 133
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LIST OF TABLES
Table 1. Summary of the Draft Freshwater Selenium Ambient Chronic Water Quality Criterion
for Protection of Aquatic Life xiv
Table 2.1. Predominant chemical forms of selenium in discharges associated with different
activities and industries 9
Table 2.2. Summary of Assessment Endpoints and Measures of Effect Used in Criteria
Derivation for Selenium 24
Table 3.1. Maternal Transfer Reproductive Toxicity Studies 40
Table 3.2. Ranked Genus Mean Chronic Values for Fish Reproductive Effects Measured as Egg
or Ovary Concentrations 42
Table 3.3. Ranked Invertebrate Whole-Body Chronic Values with Translation to Expected
Accompanying Fish Whole-Body Concentrations 46
Table 3.4. Four lowest Genus Mean Chronic Values for Fish Reproductive Effects 47
Table 3.5. Tested Reproductive-Effect Egg-Ovary (EO) Concentrations Converted to Whole-
Body (WB) Concentrations 48
Table 3.6. The lowest four reproductive-effect whole-body GMCVs 49
Table 3.7. Tested Reproductive-Effect Egg-Ovary (EO) Concentrations Converted to Muscle
(M) Concentrations 51
Table 3.8. The lowest four reproductive-effect fish muscle GMCVs 51
Table 3.9. Minimum Data Requirements Summary Table Reflecting the Number of Species and
Genus Level Mean Values Represented in the Chronic Toxicity Dataset for Selenium in
Freshwater 53
Table 3.10. EPA-derived Trophic Transfer Function (TTF) Values for Freshwater Aquatic
Invertebrates 69
Table 3.11. EPA-Derived Trophic Transfer Function (TTF) Values for Freshwater Fish 70
Table 3.12. EPA-Derived Egg-Ovary to Whole-Body Conversion Factor (CF) Values 73
Table 3.13. Data for the 53 Site Minimum Translations of the Egg-Ovary Criterion
Concentration Element to a Water Column Concentration 82
Table 3.14. Water column criterion element concentration values translated from the egg-ovary
criterion element 84
Table 3.15. Representative Values of the Intermittent Water Criterion Concentration Element. 88
Table 4.1. 2014 External Peer Review Draft Freshwater Selenium Ambient Water Quality
Chronic Criterion for Aquatic Life 91
Table 6.1. Correlation matrix (values of r) for Ictaluridae and Centrarchidae abundance and for
selenium food chain contamination for the Hyco Reservoir data reported by Crutchfield
(2000) 107
Table 6.2. Freshwater Chronic Values from Acceptable Tests - Non-Reproductive Endpoints
(Parental Females Not Exposed) 119
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LIST OF FIGURES
Figure 2.1. Map indicating deposits of selenium in mining regions 6
Figure 2.2. Areas of western U.S. susceptible to selenium contamination (light gray) and where
agricultural land is irrigated (darker green) 7
Figure 2.3. Diagram of selenium partitioning, bioaccumulation, and effects in the aquatic
environment 25
Figure 2.4. Conceptual model for translating the selenium egg-ovary concentration to a water
column concentration 31
Figure 3.1. Distribution of reproductive-effect GMCVs for fish measured as egg-ovary
concentrations 47
Figure 3.2. Distribution of reproductive-effect GMCVs for fish, measured as egg-ovary
concentrations but converted to whole-body concentrations as shown in Table 6 50
Figure 3.3. Distribution of reproductive-effect GMCVs for fish, measured as egg-ovary
concentrations but converted to muscle concentrations as shown in Table 3.7 52
Figure 3.4. Example aquatic system scenarios and the derivation of the equation parameter
TTFcomposite 61
Figure 3.5. Effect of relative sample collection time on correlation coefficients of selenium
measurements in particulate material, and invertebrate and fish tissue 66
Figure 3.6. Enrichment factors (EF) for 69 aquatic sites derived from published studies and
organized by waterbody type 77
Figure 3.7. The relationship between Cwater and Cparticuiate, and Cwater and EF for the 27 lentic and
42 lotic aquatic systems 79
Figure 3.8. Distribution of EF values for the same 69 aquatic systems as shown in Figure 3.6 and
3.7 grouped by lentic and lotic aquatic system categories 80
Figure 3.9. Probability distribution of the water column concentrations translated from the egg-
ovary criterion element at 20 lentic and 33 lotic aquatic sites 84
Figure 3.10. Illustration of intermittent spike exposure occurring for a certain percentage of time
(e.g., 10%) over a 30-day period, and background exposure occurring for the remaining
percentage of time (e.g., 90%) 87
Figure 6.1. Crutchfield (2000) observations of channel catfish (CCF) and largemouth bass
(LMB) in Hyco Reservoir beginning a few years after populations of largemouth bass
had been reduced by Se contamination 109
Figure 6.2. Distribution of (a) fish (Trophic Level 3) non-reproductive GMCVs for fish
measured as whole-body concentrations or muscle concentrations converted to whole
body, and (b) invertebrate (Trophic Level 2) GMCVs, and (c) the WB criterion
applicable to TL3 122
Figure 6.3. Distribution offish reproductive effect GMCVs from Figure 3.2 and distribution of
fish nonreproductive effect GMCVs and invertebrate GMCVs from Figure 6.2 123
Figure 6.4. Scatter plot of predicted versus measured concentrations of selenium in fish 125
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NOTICE
This draft document has undergone a contractor-led external expert peer -review process,
as well as an EPA intra-agency review process following publication and public comments
received in July 2014. Final review by the Health and Ecological Criteria Division, Office of
Science and Technology, U.S. Environmental Protection Agency, has been completed, and the
draft is approved for publication.
When published in final form, this document will provide guidance to States and Tribes
authorized to establish water quality standards under the Clean Water Act (CWA), to protect
aquatic life from toxic effects of selenium. Under the CWA, States and Tribes are to establish
water quality criteria to protect designated uses. State and tribal decision makers retain the
discretion to adopt approaches on a case-by-case basis that differ from this guidance when
appropriate. While this document contains EPA's draft scientific recommendations regarding
ambient concentrations of selenium that protect aquatic life, it does not substitute for the CWA
or EPA's regulations; nor is it a regulation itself. Thus, it cannot impose legally-binding
requirements on EPA, States, Tribes, or the regulated community, and might not apply to a
particular situation based upon the circumstances. EPA may change this draft document in the
future. This document has been approved for publication by the Office of Science and
Technology, Office of Water, U.S. Environmental Protection Agency.
Mention of trade names or commercial products does not constitute endorsement or
recommendation for use. This document can be downloaded from:
http://www.epa.gov/waterscience/criteria/aqlife.html
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FOREWORD
Section 304(a)(l) of the Clean Water Act of 1977 (P.L. 95-217) requires the
Administrator of the Environmental Protection Agency to publish water quality criteria that
accurately reflect the latest scientific knowledge on the kind and extent of all identifiable effects
on health and welfare that might be expected from the presence of pollutants in any body of
water, including ground water. This document is a second proposal of an updated chronic
ambient water quality criterion (AWQC) for the protection of aquatic life based upon
consideration of all available information relating to effects of selenium on aquatic organisms.
Comments from the general public and an external expert peer review panel on an earlier draft
published in the Federal Register in May, 2014 have been incorporated into this proposal.
The term "water quality criteria" is used in two sections of the Clean Water Act, section
304(a)(l) and section 303(c)(2). The term has a different program impact in each section. In
section 304, the term represents a non-regulatory, scientific assessment of ecological effects. The
criterion presented in this document is such a scientific assessment. If water quality criteria
associated with specific designated uses are adopted by a state as water quality standards under
section 303, and approved by EPA, they become enforceable maximum acceptable pollutant
concentrations in ambient waters within that state. Water quality criteria adopted in state water
quality standards could have the same numerical values as criteria developed under section 304.
However, states may adjust water quality criteria developed under section 304 to reflect local
environmental conditions and human exposure patterns. Alternatively, states may use different
data and assumptions than EPA in deriving numeric criteria that are scientifically defensible and
protective of designated uses. It is not until their adoption as part of state water quality standards,
and subsequent approval by EPA, that criteria become enforceable. Guidelines to assist the states
and Indian tribes in modifying the criteria presented in this document are contained in the Water
Quality Standards Handbook (U.S. EPA 1994), which along with additional guidance on the
development of water quality standards and other water-related programs of this agency have
been developed by the Office of Water.
This draft document is guidance only. It does not establish or affect legal rights or
obligations. It does not establish a binding norm and cannot be finally determinative of the issues
addressed. Agency decisions in any particular situation will be made by applying the Clean
Water Act and EPA regulations on the basis of specific facts presented and scientific information
then available.
Elizabeth Southerland
Director
Office of Science and Technology
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ACKNOWLEDGEMENTS
Technical Analysis Leads
Joseph Beaman, U.S. EPA, Office of Water, Office of Science and Technology, Health and
Ecological Criteria Division, Washington, DC
beaman.joe@epa.gov (primary contact person)
Gary Russo, U.S. EPA, Office of Water, Office of Science and Technology, Standards and
Health Protection Division, Washington, DC
russo.gary@epa.gov
Charles Delos, U.S. EPA, Office of Water, Office of Science and Technology, Health and
Ecological Criteria Division, Washington, DC (retired)
U.S. EPA Office of Water Reviewers
Elizabeth Behl
Kathryn Gallagher
Lisa Huff
Michael Elias
Intra-Agency Panel Peer Reviewers (2014-2015)
Dale Hoff and Charles Stephan, U.S. EPA, Office of Research and Development, Mid-Continent
Ecology Division, Duluth, MN
Cindy Roberts, U.S. EPA, Office of Research and Development, Office of Science Policy,
Washington, DC
Jim Lazorchak, U.S. EPA, Office of Research and Development, National Exposure Research
Laboratory, Cincinnati, OH
Jeff Gallagher, U.S. EPA, Office of Chemical Safety and Pollution Prevention, Office of
Pollution Prevention and Toxics, Washington, DC
Laura Phillips, David Hair, and Scott Wilson, U.S. EPA, Office of Water, Office of Wastewater
Management, Washington, DC
Rosaura Conde, Ruth Chemerys, and Eric Monschein, U.S. EPA, Office of Water, Office of
Wetlands, Oceans, and Watersheds, Washington, DC
Lars Wilcut and Jim Keating, U.S. EPA, Office of Water, Office of Science and Technology,
Washington, DC
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Julianne McLaughlin, ORISE Postdoctoral Participant, Office of Water, Office of Science and
Technology, Washington, DC
Cheryl Atkinson and Frank Borsuk, U.S. EPA Region 3, Philadelphia, PA, and Wheeling, WV
Joel Hansel, U.S. EPA Region 4, Atlanta, GA
Dean (Robie) Anson, Candice Bauer, and Angela Vincent, U.S. EPA Region 5, Chicago, IL
Lareina Guenzel, U.S. EPA Region 8, Denver, CO
Diane Fleck, Eugenia McNaughton, and Daniel Oros, U.S. EPA Region 9, San Francisco, CA
Lisa Macchio, and Burt Shepard, U.S. EPA Region 10, Seattle, WA
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ACRONYMS
AE Assimilation Efficiency
AWQC Ambient Water Quality Criteria
BAF Bioaccumulation Factor
CF Conversion Factor
CV Chronic Value (expressed in this document as an EC 10)
CWA Clean Water Act
dw Dry Weight
ECx Effect Concentration at X Percent Effect Level
EF Enrichment Factor
EPA Environmental Protection Agency
EO Egg Ovary
FCV Final Chronic Value
GMCV Genus Mean Chronic Value
IR Ingestion Rate
ke Rate of selenium loss
ku Rate of selenium uptake
LOEC Lowest Observed Effect Concentration
M Muscle
MATC Maximum Acceptable Toxicant Concentration (expressed mathematically as the
geometric mean of the NOEC and LOEC)
MDR Minimum Data Recommendations or Requirements
NPDES National Pollutant Discharge Elimination System
NOEC No Observed Effect Concentration
SMCV Species Mean Chronic Value
TMDL Total Maximum Daily Load
TRAP EPA's Statistical Program: Toxicity Relationship Analysis Program (Version
1.22)
TTF Trophic Transfer Factor
WB Whole body
WQBELS Water Quality-based Effluent Limitations
WQC Water Quality Criteria
WQS Water Quality Standards
ww Wet Weight
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EXECUTIVE SUMMARY
This document sets forth the basis for and derivation of the Clean Water Act, Section
304(a) water quality criterion for protecting aquatic life from harmful effects of selenium, a
naturally occurring chemical element that is nutritionally essential in small amounts, but toxic at
higher concentrations. This assessment provides a critical review of all data identified in EPA's
literature search quantifying the toxicity of selenium to aquatic organisms, and provides a basis
for a criterion that will assure protection of population assemblages offish, amphibians, aquatic
invertebrates, and plants.
Although selenium may cause acute toxicity at high concentrations, the most deleterious
effect on aquatic organisms is due to its bioaccumulative properties; these chronic effects are
found at lower concentrations than acute effects. Organisms in aquatic environments exposed to
selenium accumulate it primarily through their diet, and not directly through water (Chapman et
al. 2010). It is also recognized that selenium toxicity occurs primarily through transfer to the
eggs and subsequent reproductive effects. Consequently, in harmony with the recommendations
of expert panels (U.S. EPA 1998, Chapman et al. 2010) and with peer review and public
comments on previous U.S. EPA (2004, 2014) drafts, the Agency developed a chronic criterion
reflective of the reproductive effects of selenium concentrations on fish species.
The 2015 "Draft Aquatic Life Ambient Water Quality Criterion for Selenium -
Freshwater, 2015, is a chronic criterion that is composed of four elements. The recommended
elements are: (1) a fish egg-ovary element; (2) a fish whole-body and/or muscle element; (3) a
water column - element (one value for lentic and one value for lotic aquatic systems); and (4) a
water column intermittent element to account for potential chronic effects from repeated, short-
term exposures (one value for lentic and one value for lotic aquatic systems). The assessment of
the available data for fish, invertebrates, and amphibians indicates that a criterion value derived
from fish will protect the aquatic community. All four criterion elements applied together should
protect aquatic life from the chronic effects of exposure to total selenium in waters inhabited by
fish, as well as "fishless waters."
Because the factors that determine selenium bioaccumulation vary among aquatic
systems, site-specific water column criterion values may be necessary at aquatic sites with high
selenium bioaccumulation to ensure adequate protection of aquatic life (Appendix K). Finally,
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the freshwater chronic selenium criterion applies only to aquatic life, and is not intended to
address selenium toxicity to aquatic-dependent wildlife such as aquatic-dependent birds.
The toxicity studies relevant to the derivation of the fish tissue selenium criterion
elements involve (a) extended-duration dietary exposure, and (b) measurement of total selenium
in the tissue of the target organism. Selenium either in fish whole-body or in muscle is usually
measured in non-reproductive studies and selenium in eggs or ovaries is typically measured in
reproductive studies. Selenium accumulation in the eggs of the exposed adult female prior to
spawning has been shown to yield the most robust relationship (statistically significant) with
occurrence of deformities and reduced survival of the offspring.
The outcome of assessing both reproductive and non-reproductive studies under
laboratory and field conditions led EPA to the conclusion, consistent with expert consensus
(Chapman et al. 2009, 2010), that reproductive effects, linked to egg-ovary selenium
concentrations, provide a sound basis for the criterion compared to non-reproductive (e.g.,
survivorship, growth) endpoints. Reproductive effects have been linked to observed reductions in
the populations of sensitive fish species in waterbodies having elevated concentrations of
selenium (Young et al. 2010). We applied the species sensitivity distribution concepts from the
U.S. EPA Guidelines for Deriving Numerical National Water Quality Criteria for the Protection
of Aquatic Organisms and their Uses (Stephan et al. 1985), to derive the selenium criterion.
Based on the available data, expressed as ECio values, the draft egg-ovary criterion element is
15.8 milligrams selenium per kilogram dry weight (mg Se/kg dw), based primarily on 16
reproductive studies representing ten fish genera. All other selenium criterion elements in the
draft document are derived from this egg-ovary criterion element.
EPA recommends states and tribes adopt all four elements of the criterion into their water
quality standards. Two elements are based on the concentration of selenium in fish tissue (eggs
and ovaries, and whole-body or muscle) and two elements are based on the concentration of
selenium in the water column (a 30-day chronic element and an intermittent exposure element).
Both water column elements are further refined into values for lentic waters (e.g., lakes and
impoundments) and lotic waters (e.g., rivers and streams). The difference between lentic and
lotic water column values reflect the observed difference in selenium bioaccumulation in these
two categories of aquatic systems. EPA derived the intermittent element based on the chronic 30-
day water column element and the fraction of any 30-day period during which elevated selenium
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concentrations occur. EPA is recommending the intermittent value to address short-term
exposures that contribute to chronic effects through selenium bioaccumulation (e.g., stormwater
overflows from storage ponds or other intermittent releases). EPA derived the values for the
water-column criterion elements from the egg-ovary element by assessing food-chain
bioaccumulation based on available data collected at lentic and lotic systems in the continental
United States. Thus, all four criterion elements are based on reproductive effects in freshwater
fish.
EPA primarily used field studies to assess selenium bioaccumulation in particulate
material (algae, detritus, and sediment) and used field observations and laboratory data to
quantify and model the trophic transfer of selenium from particulate material into invertebrates,
and from invertebrates into fish. EPA additionally used field and laboratory observations to
assess species-specific selenium partitioning between different tissues within a fish (whole-body,
eggs and/or ovaries, and muscle). EPA validated this approach using selenium measurements
from aquatic systems with a range of bioaccumulation potential.
While more than half the available chronic studies were fish studies, available field data
and laboratory toxicity studies suggest that a criterion based on fish will protect amphibians,
aquatic invertebrates, and plants since these taxa appear to be less sensitive than fish (see
Sections 3.1.4 and 6.1.4).
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Table 1. Summary of the Draft Freshwater Selenium Ambient Chronic Water Quality
Criterion for Protection of Aquatic Life.
(See Section 4 for the complete criterion statement.)
Media
Type
Fish Tissue
Water Column
Criterion
Element
Egg/Ovary1
Fish Whole
Body or
Muscle2
Monthly
Average
Exposure
Intermittent Exposure4
Magnitude
15.8mg/kg
8.0 mg/kg
whole body
or
11.3 mg/kg
muscle
(skinless,
boneless filet)
1.2 jig/Lin
lentic aquatic
systems
3.1 jig/Linlotic
aquatic systems
WQCint =
— C
bkgrnd
/ int)
I int
Duration
Instantaneous
measurement5
Instantaneous
measurement5
30 days
Number of days/month with
an elevated concentration
Frequency
Never to be
exceeded
Never to be
exceeded
Not more than
once in three
years on
average
Not more than once in three
years on average
Overrides any whole-body, muscle, or water column elements when fish egg/ovary
concentrations are measured, except in certain situations. See footnote 3.
r\
Overrides any water column element when both fish tissue and water concentrations are
measured, except in certain situations. See footnote 3.
3 Water column values are based on dissolved total selenium (includes all oxidation states, i.e.,
selenite, selenate, organic selenium and any other forms) in water. Water column values have
primacy over fish tissue values under two circumstances: 1) "Fishless waters" (waters where
fish have been extirpated or where physical habitat and/or flow regime cannot sustain fish
populations); and 2) New (see glossary) or increased inputs of selenium from a specific source
until equilibrium is reached.
4 Where WQCso-dayis the water column monthly element, for either a lentic or lotic system, as
appropriate. Cbkgrnd is the average background selenium concentration, and f;nt is the fraction of
any 30-day period during which elevated selenium concentrations occurs, with f;nt assigned a
value >0.033 (corresponding to 1 day). See Section 3.3.
5 Instantaneous measurement. Fish tissue data provide point measurements that reflect integrative
accumulation of selenium over time and space in the fish at a given site. Selenium concentrations
in fish tissue are expected to change only gradually over time in response to environmental
fluctuations.
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The chronic selenium criterion is derived to be protective of the entire aquatic
community, including fish, amphibians, and invertebrates. Because fish are the most sensitive to
selenium effects, selenium water quality criterion elements based on fish tissue (egg-ovary,
whole body, and/or muscle) sample data override the criterion elements based on water column
selenium data due to the fact, noted above, that fish tissue concentrations provide the most robust
and direct information of potential selenium effects in fish. However, because selenium
concentrations in fish tissue are a result of selenium bioaccumulation via dietary exposure, there
are two specific circumstances where the fish tissue concentrations do not fully represent
potential effects on fish and the aquatic ecosystem: 1) In "fishless" waters, and 2) areas with new
or increased selenium inputs.
Fishless waters are defined as waters with insufficient instream habitat and/or flow to
support a population of any fish species on a continuing basis, or waters that once supported
populations of one or more fish species but no longer support fish (i.e., extirpation) due to
temporary or permanent changes in water quality (e.g., due to selenium pollution), flow or
instream habitat. Because of the inability to collect sufficient fish tissue to measure selenium
concentrations in fish in such waters, water column concentrations will best represent selenium
levels required to protect aquatic communities and downstream waters in such areas.
New inputs are defined as new activities resulting in selenium being released into a lentic
or lotic waterbody. Increased input is defined as an increased discharge of selenium from a
current activity released into a lentic or lotic waterbody. New or increased inputs will likely
result in increased selenium in the food web, likely resulting in increased bioaccumulation of
selenium in fish over a period of time until the new or increased selenium release achieves a
quasi-"steady state" balance within the food web. EPA estimates that the concentration of
selenium in fish tissue will not represent a "steady state" for several months in lotic systems, and
longer time periods (e.g., 2 to 3 years) in lentic systems, dependent upon the hydrodynamics of a
given system; the location of the selenium input related to the shape and internal circulation of
the waterbody, particularly in reservoirs with multiple riverine inputs; and the particular food
web. Estimates of time to achieve steady state under new or increased selenium input situations
are expected to be site dependent, so local information should be used to better refine these
estimates for a particular waterbody. Thus, EPA recommends that fish tissue concentration not
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override water column concentration until these periods of time have passed in lotic and lentic
systems, respectively.
EPA recommends that states and tribes adopt into their water quality standards a
selenium criterion that expresses the four elements as a single criterion composed of multiple
parts in a manner that explicitly affirms the primacy of the whole-body or muscle element over
the water column elements, and the egg-ovary element over any other element. Adopting the fish
whole-body or muscle tissue element into water quality standards ensures the protection of
aquatic life when measurements from fish eggs or ovary are not available, and adopting the water
column element ensures protection when fish tissue measurements are not available (see Section
3). EPA recommends that when states implement the criterion for selenium under the National
Pollutant Discharge Elimination System (NPDES) permits program, states should establish
additional procedures to facilitate translation of the fish tissue criterion concentration elements
into water concentration permit limits.
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1 INTRODUCTION AND BACKGROUND
The objective of the Clean Water Act (CWA) is to "restore and maintain the chemical,
biological and physical integrity of the Nation's waters." One of the tools that EPA uses to meet
this objective is the development of ambient water quality criteria (AWQC) under section
304(a)(l) of the Act. As provided for by the Clean Water Act, EPA reviews and from time to
time revises 304(a) AWQC to ensure the criteria are consistent with the latest scientific
information. Section 304(a) aquatic life criteria serve as recommendations to states and tribes
for defining ambient water concentrations that will protect against adverse ecological effects to
aquatic life resulting from exposure to a pollutant found in water from direct contact, ingestion of
contaminated water and/or food. Aquatic life criteria address the Clean Water Act goals of
providing for the protection and propagation offish and shellfish. When adopted into state water
quality standards (WQS), these criteria can become a basis for establishing National Pollutant
Discharge Elimination System (NPDES) program permit limits, thresholds for listing impaired
waters [Section 303(d)] and Total Maximum Daily Loads (TMDLs).
1.1 HISTORY OF THE EPA SELENIUM AWQC FOR AQUATIC LIFE
In 1980 EPA first published numeric aquatic life criteria for selenium in freshwater.
These criteria were based on water-only exposure (no dietary exposure). In order to address the
lack of consideration of bioaccumulation in the 1980 selenium criteria, in 1987 EPA published
updated selenium criteria to address field-based toxicity observed in aquatic ecosystems at levels
below the existing criteria values. The 1987 criteria were field-based and accounted for both the
water column and dietary uptake pathways manifested at Belews Lake, North Carolina (USA), a
cooling water reservoir where water quality and fish communities had been affected by selenium
loads from a coal-fired power plant. At that time EPA also provided an acute criterion of 20 |ig/L
derived from a reverse application of an acute-chronic ratio obtained from conventional water-
only exposure toxicity tests applied to the 5 |ig/L chronic value based on dietary and water
column exposure in Belews Lake.
In 1998-1999 EPA published a revised acute criterion, a formula that recognized that the
two oxidation states, selenate and selenite, appeared to have substantially different acute
toxicities. This acute criterion assumed toxicity was based on water-only exposure. Subsequent
research has demonstrated that sulfate levels influence selenate toxicity in water-only exposures.
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In 1998 EPA held a peer consultation workshop (EPA-822-R-98-007) to evaluate new
science available for selenium relevant to the selenium aquatic life criterion. EPA concluded, and
the peer reviewers agreed, that fish-tissue values more directly represent chronic adverse effects
of selenium than the conventional water concentration approach used by EPA to protect aquatic
life, because chronic selenium toxicity is primarily based on the food-chain bioaccumulation
route, not on a water route of exposure.
In 2004 EPA published a draft chronic whole-body fish-tissue criterion with a water-
based monitoring trigger in the summer and fall. The critical effect considered at that time was
the impact on survivorship based on overwintering stress to bluegill sunfish. An acute criterion
was estimated at that time that addressed concerns with the species of selenium present and
adjusted for sulfate levels; however, it did not address the dietary uptake pathway.
Further refinement of the fish tissue approach occurred in 2009 based on the findings of a
Pellston scientific workshop on the ecological risk assessment of selenium (Chapman et al. 2009,
2010). As presented by Chapman et al. (2009), some key findings resulting from that workshop
are:
• Diet is the primary pathway of selenium exposure for both invertebrates and vertebrates.
• Traditional methods for predicting toxicity on the basis of exposure to dissolved [water
column] concentrations do not work for selenium because the behavior and toxicity of
selenium in aquatic systems are highly dependent upon site-specific factors, including
food web structure and hydrology.
• Selenium toxicity is primarily manifested as reproductive impairment due to maternal
transfer, resulting in embryotoxicity and teratogenicity in egg-laying vertebrates.
In this 2015 draft freshwater chronic criterion for selenium, EPA includes revisions based
on the public and external expert peer reviews in 2014, data and information from additional
studies provided by the public and peer reviewers, and additional scientific analyses. EPA also
conducted a new literature review and reanalyzed data considered in the 2004 and 2009 draft
criteria documents. This draft criterion reflects the latest scientific consensus (e.g., Chapman et
al. 2010) on the reproductive effects of selenium on aquatic life and their measurement in aquatic
systems and supersedes all previous national aquatic life water quality criteria for selenium.
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EPA is recommending a national selenium criterion expressed as 4 elements. All
elements are protective against chronic selenium effects, and account for both short term and
longer term exposure to selenium. Two elements are based on the concentration of selenium in
fish tissue (eggs and ovaries, and whole-body or muscle) and two elements are based on the
concentration of selenium in the water-column (two 30-day chronic values and an intermittent
value). EPA derived the 30-day chronic water column element from the egg-ovary element by
modeling selenium bioaccumulation in food webs of lotic and lentic aquatic systems. EPA is
recommending the intermittent value to address short-term exposures that could contribute to
chronic effects through selenium bioaccumulation in either lotic or lentic systems. EPA derived
the intermittent element based on the chronic 30-day water column element and the fraction of
any 30-day period during which elevated selenium concentrations occur. These water column
criterion elements apply to the total of all oxidation states (selenite, selenate, organic selenium,
and any other forms) (See Appendix L for Analytical Methods for Measuring Selenium).
Aquatic communities are expected to be protected by this chronic criterion from any potential
acute effects of selenium.
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2 PROBLEM FORMULATION
Problem formulation provides a strategic framework for water quality criteria
development by focusing the effects assessment on the most relevant chemical properties and
endpoints. The structure of this effects assessment is consistent with EPA's Guidelines for
Ecological Risk Assessment (U.S. EPA 1998).
This ecological effects assessment defines a scientifically-defensible water quality
criterion for selenium under section 304(a)(l) of the Clean Water Act. Clean Water Act Section
304(a)(l) requires EPA to develop criteria for water quality that accurately reflect the latest
scientific knowledge. These criteria are based solely on data and best professional scientific
judgments on toxicological effects. Criteria are developed following overarching guidance
outlined in the Agency's Guidelines for Deriving Numerical National Water Quality Criteria for
the Protection of Aquatic Organisms and Their Uses (Stephan et al. 1985), hereafter referred to
as "U.S. EPA Ambient Water Quality Criteria Guidelines". States and authorized tribes may
adopt EPA's recommended criteria into their water quality standards to protect designated uses
of water bodies, they may modify EPA's criteria to reflect site-specific conditions, or they may
derive criteria using other scientifically-defensible methods, all subject to EPA review and
approval.
2.1 OVERVIEW OF SELENIUM SOURCES AND OCCURRENCE
Selenium is a naturally occurring element present in sedimentary rocks and soils. It is
also present in the aquatic environment as methyl derivatives of selenium, naturally occurring in
freshwaters through methylation by bacteria (Ranjard et.al. 2003). There are around 40 known
selenium-containing minerals, some of which can have as much as 30% selenium, but all are rare
and generally occur together with sulfides of metals such as copper, zinc and lead (Emsley
2011). The distribution of organic-enriched, sedimentary shales, petroleum source rocks, ore
deposits, phosphorites, and coals, in which selenium typically co-occurs, is well characterized in
the United States (Presser et al. 2004) (see Figure 2.1 and Figure 2.2). Two major
anthropogenic activities cause selenium mobilization and introduction into aquatic systems. The
first is the mining of metals, minerals and refinement and use of fossil fuels; the second is
irrigation of selenium-rich soils.
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Mining activities bring selenium-enriched deposits to the surface, where they are exposed
to physical weathering processes (Error! Reference source not found.). The release of selenium
related to resource extraction activities is most common in the phosphate deposits of southeast
Idaho and adjacent areas of Wyoming, Montana, and Utah, and in coal mining areas in portions
of West Virginia, Kentucky, Virginia, and Tennessee (Presser et al. 2004). Where selenium-
containing minerals, rocks, and coal are mined, selenium can be mobilized when rock
overburden and waste materials are crushed, increasing the surface area and exposure of material
to weathering processes. Selenium contamination of surface waters can also occur when sulfide
deposits of iron, uranium, copper, lead, mercury, silver, and zinc are released during the mining
and smelting of these metal ores. Where coal is burned for power production, selenium can enter
surface waters as drainage from fly-ash ponds and fly-ash deposits on land (Gillespie and
Baumann 1986). Fly ash deposits have a high surface area to volume ratio, resulting in rates of
selenium in leachate several times higher than from the parent feed coal (Fernandez-Turiel et al.
1994). The refining of crude oil containing high levels of selenium can also be a major source of
loading in certain water bodies (Maher et al. 2010).
Irrigation of selenium-rich soils for crop production in arid and semi-arid regions of the
country (Figure 2.2) can mobilize selenium and move it off-site in surface water runoff or via
leaching into ground water. Where deposits of Cretaceous marine shales occur, they can weather
to produce high selenium soils; such soils are present in many areas of the western U.S. (Lemly
1993c). Selenium is abundant in the alkaline soils of the Great Plains, and some ground waters in
California, Colorado, Kansas, Oklahoma, South Dakota and Wyoming contain elevated
concentrations of selenium due to weathering of and leaching from rocks and soils. In semi-arid
areas of the West, irrigation water applied to soils containing soluble selenium can leach
selenium. The excess water (in tile drains or irrigation return flow) containing selenium can be
discharged into basins, ponds, or streams. For example, elevated selenium levels at the Kesterson
Reservoir in California originated from agricultural irrigation return flow collected in tile drains
that discharged into the reservoir (Ohlendorf et al. 1986).
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Figure 2.1. Map indicating deposits of selenium in mining regions.
Light shading indicates lower selenium concentrations (< 7.2 mg/L), whereas darker shading
indicates higher selenium concentrations (> 7.2 mg/L) in underlying geology. Source of Map:
SAIC, 2008.
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a 1
0 2SO 500 KILOMETERS
|
Uppor C
r than 2 5 on<1 wtwre gpatoffr iirtis .in? ma Inly
Kis. oc Tertian mar1m> wdtiwrnary deposit*
LI :," -I -i i M •.•IKli il I -I, I
Figure 2.2. Areas of western U.S. susceptible to selenium contamination (light gray) and
where agricultural land is irrigated (darker green).
Overlap of light gray and darker green show areas susceptible to selenium discharge from
irrigation. Note: Eastern U.S. is not as susceptible since selenium does not occur at the surface
where agricultural practices can mobilize selenium. Source of Map: Seilor et al. 1999 page 31.
Atmospheric emissions of selenium can originate from several sources including power
plants and other facilities that burn coal or oil, selenium refineries that provide selenium to
industrial users, base metal smelters and refineries, resource extraction industries, milling
operations, and end-product manufacturers (e.g., semiconductor manufacturers) (ATSDR 2003).
Airborne selenium particles can settle either on surface waters or on soils from which selenium
can be further transported and deposited into water bodies through ground or surface water
conveyances or runoff.
The chemical form of selenium that dominates a location is usually dependent on its
sources, effluent treatments, and biogeochemical processes in the receiving waters. Irrigation
/^
activities in areas with seleniferous soils typically mobilize selenate (SeC>4 or Se[VI]) (Seller et
/^
al. 2003). Combustion of coal for power generation creates predominantly selenite (SeOs , or
Se[IV]) in the fly ash waste due to the temperatures, pH, and redox conditions involved with the
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process (Huggins et al. 2007). Similar conditions during refinement of crude oil can also result in
high concentrations of selenite relative to selenate, as was observed in the San Francisco Bay
estuary in the 1980s (Cutter 1989). Although selenite is the dominant species in the discharges
resulting from crude oil refining and coal burning using conventional technologies, the
implementation of alternative treatment technologies can alter the relative concentrations of
selenate and selenite. For example, in scrubbers with forced oxidation systems that produce
strong oxidizing conditions and high temperatures, the majority of discharged selenium is in the
form of selenate (Maher et al. 2010). Table 2.1 shows the predominant form of selenium that is
associated with different activities and industries.
U.S. EPA's Office of Water and Office of Research and Development conducted the first
statistically based survey of contaminants in fish fillets from U.S. Rivers from 2008 through
2009. This national fish survey was conducted under the framework of U.S. EPA's National
Rivers and Streams Assessment (NRSA), a probability-based survey designed to assess the
condition of the Nation's streams and rivers (Lazorchak et al. 2014). During June through
October of 2008 and 2009, field teams applied consistent methods nationwide to collect samples
offish species commonly consumed by humans at 542 randomly selected river locations (> 5th
order based on l:100,000-scale Strahler order) in the lower 48 states. They collected one
composite fish sample at every sampling location, with each composite consisting of five
similarly sized adult fish of the same species from a list of target species. Largemouth and
smallmouth bass were the primary species collected for the study, accounting for 34% and 24%
of all fish composites, respectively. Samples were collected from both non-urban (379 sites) and
urban locations (163 sites). Each fillet composite sample was homogenized and analyzed using
an ICP-MS (Inductively Coupled Plasma- Mass Spectrometry) method for total selenium, and
results were reported as wet weight. Three of the 542 samples (approximately 0.6%) exceeded
the 2015 draft criterion for muscle tissue, 11.3 mg/kg dw. The maximum value detected was
17.75 mg Se/kg dw muscle, the median was 1.86 mg Se/kg dw, and the minimum 0.24 mg Se/kg
dw. Concentrations in urban and non-urban sites did not differ markedly.
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Table 2.1. Predominant chemical forms of selenium in discharges associated with different
activities and industries.
Selenium Form
Selenate
Selenite
Organoselenium
Sources
Agricultural irrigation drainage
Treated oil refinery effluent
Mountaintop coal mining/ valley fill leachate
Copper mining discharge
Oil refinery effluent
Fly ash disposal effluent
Phosphate mining overburden leachate
Treated agricultural drainage (in ponds or lagoons)
Source: Presser and Ohlendorf 1987; Zhang and Moore 1996; Cutter and Diego-McGlone 1990.
2.2 ENVIRONMENTAL FATE AND TRANSPORT OF SELENIUM IN THE AQUATIC
ENVIRONMENT
The fate and transport of selenium in aquatic systems is affected by the distribution of
selenium species and their transformations in water, sediment, and biota. These transformations
include the assimilation and conversion of inorganic selenium to organic selenium species in
plants and microbes that are transferred to higher trophic level consumer species throughout the
aquatic food web.
2.2.1 Selenium Species in Aquatic Systems
Aquatic organisms are exposed to a combination of predominantly organic selenium
species present in the food web throughout their life history; reproductive effects integrate these
exposures, to transformed inorganic and organic species of selenium. The bioavailability and
toxicity of selenium depend on both its concentration and speciation (Cutter and Cutter 2004;
Meseck and Cutter 2006; Reidel et al. 1996). Selenium exists in four oxidation states (VI, IV, 0,
II) and in a wide range of chemical and physical species across these oxidation states (Doblin et
al. 2006; Maher et al. 2010; Meseck and Cutter 2006). Therefore, in the effects assessment that
follows, we have correlated the adverse effects on aquatic life with total dissolved selenium.
In oxygenated surface waters, the primary dissolved selenium species are selenate
(SeC>42 or Se[VI]) and selenite (SeOs2 , or Se[IV]), as well as dissolved organic selenides (-II)
formed from fine particulate organic matter (e.g., Doblin et al. 2006; Meseck and Cutter 2006).
The relative abundance of selenate and selenite depends on relative contributions from the
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geologic and anthropogenic sources of selenium to the receiving waters, as there is negligible
inter-conversion between the two species (e.g., Maher et al. 2010). Aqueous selenite is more
abundant than selenate when the majority of selenium originates from discharges from coal fly
ash tailings or oil refineries (e.g., Cutter 1989; Huggins et al. 2007). Particulate species in the
water column include selenate, selenite, and elemental selenium (Se(0)) bound to resuspended
sediments and organic particles, as well as particulate organic selenium species incorporated into
suspended detritus (e.g., Cutter and Bruland 1984; Meseck and Cutter 2006).
In sediments, selenate and selenite can be reduced to iron selenides or elemental selenium
under abiotic or biotic processes; elemental selenium and selenides can be converted to selenate
under oxidizing conditions (Maher et al. 2010). For example, selenate can be reduced to
elemental selenium in sediments (e.g., Oremland 1990) in the presence of iron oxides (Chen et
al. 2008) and iron sulfides (Breynaert et al. 2008). Elemental selenium and organic selenides are
produced by selenate-reducing microbes in sediments. Overall, the reduction of selenate and
particularly selenite in sediments increases with increasing sediment organic matter (Tokunaga et
al. 1997). Selenite in particular is readily bound to iron and manganese oxy-hydroxides (Maher
et al. 2010), and is readily adsorbed to inorganic and organic particles, particularly at a lower pH
range (e.g., McLean and Bledsoe 1992; Tokungawa et al. 1997). Microbial reduction of selenite
to organic forms (via methylation) increases the solubility and bioavailability of selenium
(Simmons and Wallschlagel 2005). Plants and algae produce volatile selenium species by
biomethylation of excess selenium which, upon reaching the sediment, can be transformed to a
more bioavailable species, or deposited in the sediments and effectively removed from the
system (Diaz et al. 2009). Depending on environmental conditions, the reduction processes
described above are largely reversible, as elemental selenium and selenides in sediments can be
oxidized to selenate through microbial or abiotic transformations (e.g., Maher et al. 2010;
Tokunaga et al. 1997).
The most important transformation of selenium, with respect to its toxicity to aquatic
organisms, is in the uptake of dissolved inorganic selenium into the tissues of primary producers
at the base of the food web. The main route of entry of selenium into aquatic foodwebs is from
the consumption of particulate selenium of primary producers, and to a lesser degree, from the
consumption of sediments (Doblin et al. 2006; Luoma and Presser 2009). For algae, selenite and
organic selenides are similarly bioavailable, and both dissolved species are more bioavailable
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than selenate (e.g., Baines et al. 2001; Luoma et al. 1992). In vascular plants, selenate uptake is
greater than for the other dissolved species, as the majority of selenium uptake occurs in the
roots, and selenate is more easily transported to the shoots and leaves than selenite or organic
selenides (Dumont 2006). Following uptake, selenium is metabolized into a variety of organic
species that are assimilated into plant tissues. Selenium metabolism in plants is analogous to
sulfur metabolism (e.g., Dumont et al. 2006; Ouerdane et al. 2013). Selenate is reduced to
selenite, which is then reduced to selenide in a process involving reduced glutathione (Dumont et
al. 2006). Selenide is converted to selenocysteine (SeCys), which is then converted to
selenomethionine (SeMet) (Dumont et al. 2006). In addition to SeCys and SeMet, a variety of
other organic selenium species can be formed; however, SeCys, and particularly SeMet are
lexicologically important because these amino acids nonspecifically replace cysteine and
methionine in proteins, and are more bioavailable to higher trophic level consumers (Fan et al.
2002; Freeman et al. 2006).
2.2.2 Bioaccumulation of Selenium in Aquatic Systems
Dissolved selenium uptake by animals is slow, whatever the form, such that under
environmentally relevant conditions, dissolved selenium in the water column makes little or no
direct contribution to bioaccumulation in animals (Lemly 1985a; Ogle and Knight 1996), but
does influence the concentration of selenium in particulate matter. Selenium bioaccumulation in
aquatic organisms occurs primarily through the ingestion of food (Fan et al. 2002; Luoma et al.
1992; Maher et al. 2010; Ohlendorf et al. 1986; Presser and Ohlendorf 1987; Presser et al. 1994;
Saiki and Lowe 1987). However, unlike other bioaccumulative contaminants such as mercury,
the single largest step in selenium accumulation in aquatic environments occurs at the base of the
food web where algae and other microorganisms accumulate selenium from water by factors
ranging from several hundred to tens of thousands (Luoma and Presser 2009; Orr et al. 2012;
Stewart et al. 2010). Bioaccumulation and transfer through aquatic food webs are the major
biogeochemical pathways of selenium in aquatic ecosystems. Dissolved selenium oxyanions
(selenate, selenite) and organic selenides are assimilated into the tissues of aquatic primary
producers (trophic level 1 organisms), such as periphyton, phytoplankton, and vascular
macrophytes; and subsequently biotransformed into organoselenium. These organisms, together
with other particle-bound selenium sources, constitute the particulate selenium fraction in the
water column. Selenium from this particulate fraction is then transferred to aquatic primary
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consumers such as zooplankton, insect larvae, larval fish, and bivalves (trophic level 2), and then
to predators such as fish and birds (trophic level 3 and above). In addition to the water
concentration of selenium, the process of selenium bioaccumulation in aquatic life residing in
freshwater systems depends on several factors specific to each aquatic system. These factors
include:
Water residence time. Residence time is a measure of the average time a water molecule
will spend in a specified region of space. Residence time influences both the proportion of
selenium found in particulate and dissolved forms and the predominant form of selenium.
Organisms in waters with long residence times such as lakes, ponds, reservoirs, wetlands or
estuaries will tend to bioaccumulate more selenium than those living in waters with shorter
residence times such as rivers and streams (ATSDR 2003; EPRI2006; Luoma and Rainbow
2005; Orr et al. 2006; Simmons and Wallschlagel 2005). Several interrelated factors underlie
selenium's greater bioaccumulation potential in slow moving systems, such as food web
complexity and the organic content and reduction/oxidation potential of sediments. Finally,
selenium toxicity in flowing waters with shorter residence times may only be apparent far
downstream of their selenium sources, whereas waters with longer residence times are more
likely to exhibit selenium toxicity near their sources (Presser & Luoma 2006).
Distribution of selenium between particulate and dissolved forms. Selenium is found in
both particulate and dissolved forms in water, but direct transfer of selenium from water to
animals is only a small proportion of the total exposure. The proportion of selenium found in
particulate matter (algae, detritus, and sediment) is important because it is the primary avenue for
selenium entering into the aquatic food web (Luoma et al. 1992; Luoma and Rainbow 2005;
Ohlendorf et al. 1986; Presser and Ohlendorf 1987; Presser et al. 1994; Presser & Luoma 2006;
Saiki and Lowe 1987).
Bioaccumulation in prey. Trophic level 1 organisms such as periphyton and
phytoplankton, as well as other forms of particulate material containing selenium, such as
detritus and sediment, are ingested by trophic level 2 organisms such as mollusks, planktonic
crustaceans, and many insects, increasing the concentration of selenium in the tissues of these
higher-level organisms. Differences in the physiological characteristics of these organisms result
in different levels of bioaccumulation. Also, selenium effects on invertebrates typically occur at
concentrations higher than those that elicit effects on the vertebrates (e.g., fish and birds) that
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prey upon them. Additionally, mollusks such as mussels and clams can accumulate selenium to a
much greater extent than planktonic crustaceans and insects (although the levels do not seem to
be toxic to the mussels) due to higher ingestion rates of both particulate-bound (algae) and
dissolved selenium from the water column through filter feeding, as well as the lower rate at
which they eliminate selenium (Luoma and Rainbow 2005; Stewart et al. 2013). Because egg-
laying (oviparous) vertebrates such as fish and birds are most sensitive to selenium effects, (Janz
et al. 2010), these vertebrate consumers are also the most vulnerable groups to selenium
poisoning and the focal point of most selenium environmental assessments (Ogle and Knight
1996; Stewart etal. 2010).
Trophic transfer to predators. Bioaccumulation of selenium by higher trophic level
organisms, such as trophic level 3 and 4 fish, is highly influenced by the food web of the aquatic
environment. For example, fish that primarily consume freshwater mollusks will exhibit greater
selenium bioaccumulation than fish that consume primarily insects or crustaceans from waters
with the same concentration of dissolved selenium because mollusks tend to accumulate
selenium at higher concentrations than other trophic level 2 organisms, as noted above (Luoma
and Presser 2009; Stewart et al. 2004).
2.3 MODE OF ACTION AND TOXICITY OF SELENIUM
Selenium is a naturally occurring chemical element that is also an essential micronutrient.
Trace amounts of selenium are required for normal cellular function in almost all animals.
However, excessive amounts of selenium can also have toxic effects, with selenium being one of
the most toxic of the biologically essential elements (Chapman et al. 2010). Egg-laying
vertebrates have a lower tolerance than do mammals, and the transition from levels of selenium
that are biologically essential to those that are toxic occurs across a relatively narrow range of
exposure concentrations (Luckey and Venugopal 1977; USEPA 1987, 1998; Haygarth 1994;
Chapman et al. 2009, 2010). Selenium consumed in the diet of adult female fish is deposited in
the eggs, when selenium replaces sulfur in vitellogenin, which is transported to the ovary and
incorporated into the developing ovarian follicle (Janz et al. 2010), the primary yolk precursor.
Selenium is a member of the sulfur group of nonmetallic elements, and consequently, the
two chemicals share similar characteristics. Selenium can replace sulfur in two amino acids, the
seleno-forms being selenomethionine and selenocysteine. It has been a long-standing hypothesis
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that the cause of malformations in egg-laying vertebrates is due to the substitution of selenium
for sulfur in these amino acids and their subsequent incorporation into proteins, which causes
disruption of the structure and function of the protein. When present in excessive amounts,
selenium is erroneously substituted for sulfur, resulting in the formation of a triselenium linkage
(Se-Se-Se) or a selenotrisulfide linkage (S-Se-S), either of which was thought to prevent the
formation of the normal disulfide chemical bonds (S-S). The end result was thought to be
distorted, dysfunctional enzymes and protein molecules that impaired normal cellular
biochemistry (Diplock and Hoekstra 1976; Reddy and Massaro 1983; Sunde 1984).
Recent research, however, suggests that selenium's role in oxidative stress plays a role in
embryo toxicity, whereas selenium substitution for sulfur does not. The substitution of
selenomethionine for methionine does not appear to affect either the structure or function of
proteins (Yuan et al. 1998; Mechaly et al. 2000; Egerer-Sieber et al. 2006). The reason is
apparently due to selenium not being distally located in selenomethionine, which insulates the
protein from an effect on its tertiary structure. Although the incorporation of selenomethionine
into proteins is concentration-dependent (Schrauzer 2000), selenocysteine's incorporation into
proteins is not (Stadtman 1996). This suggests that neither selenomethionine nor selenocysteine
affect protein structure or function. In fact, Se as an essential micronutrient is incorporated into
functional and structural proteins as selenocysteine.
The role of selenium-induced oxidative stress in embryo toxicity and teratogenesis
appears to be related to glutathione homeostasis. A review of bird studies by Hoffman (2002)
showed exposure to selenium altered concentrations and ratios of reduced to oxidized glutathione
thereby increasing measurements of oxidative cell damage. Palace et al. (2004) suggested
oxidative stress due to elevated selenium levels results in pericardial and yolk sac edema in
rainbow trout embryos. Evidence for the role of oxidative stress in selenium toxicity is growing,
but mechanistic studies are needed to better understand its effects on egg-laying vertebrates. For
a more in depth discussion on the mechanism of toxicity at the cellular level including the
evidence against sulfur substitution as a cause and the role of oxidative stress, see Janz et al.
(2010).
The most well-documented, overt and severe toxic symptoms in fish are reproductive
teratogenesis and larval mortality. Egg-laying vertebrates appear to be the most sensitive taxa,
with toxicity resulting from maternal transfer to eggs. In studies involving young organisms
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exposed through transfer of selenium from adult female fish into their eggs, the most sensitive
diagnostic indicators of selenium toxicity in vertebrates occur when developing embryos
metabolize organic selenium that is present in egg albumen or yolk. It is then further metabolized
by larval fish after hatching.
A variety of lethal and sublethal deformities can occur in the developing fish exposed to
selenium, affecting both hard and soft tissues (Lemly 1993b). Developmental malformations are
among the most conspicuous and diagnostic symptoms of chronic selenium poisoning in fish.
Terata are permanent biomarkers of toxicity, and have been used to identify impacts of selenium
on fish populations (Maier and Knightl994; Lemly 1997b). Deformities in fish that affect
feeding or respiration can be lethal shortly after hatching. Terata that are not directly lethal, but
distort the spine and fins, can reduce swimming ability, and overall fitness. Because the rate of
survival of deformed young would be less than that for normal young, the percentage of
deformed adults observed during biosurveys will likely understate the underlying percentage of
deformed young, although quantitation of the difference is ordinarily not possible.
In summary, the most sensitive indicators of selenium toxicity in fish larvae are effects
modulated through the reproductive process and exhibited in fish larvae as teratogenic
deformities such as skeletal, craniofacial, and fin deformities, and various forms of edema that
result in mortality (Lemly 2002). The toxic effect generally evaluated is the reduction in the
number of normal healthy offspring compared to the starting number of eggs. In studies of young
organisms exposed to selenium solely through their own diet (rather than via maternal transfer),
reductions in survival and/or growth are the effects that are generally evaluated.
2.4 NARROW MARGIN BETWEEN SUFFICIENCY AND TOXICITY OF SELENIUM
Selenium has a narrow range encompassing what is beneficial for biota and what is
detrimental. Selenium is an essential nutrient that is incorporated into functional and structural
proteins as selenocysteine and selenomethionine. Several of these proteins are enzymes that
provide cellular antioxidant protection. Selenomethionine is readily oxidized, and its antioxidant
activity arises from its ability to deplete reactive oxygen species. Selenomethionine is required
as a mineral cofactor in the biosynthesis of glutathione peroxidases. All of the classic glutathione
peroxidases contain selenium and are found to be involved in the catalytic reaction of these many
enzymes (Allan 1999). The major function of the glutathione peroxidases involves the reduction
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of hydrogen peroxide to water at the expense of the oxidation of glutathione, the enzyme's
cofactor, an important antioxidant process at normal dietary levels.
Aquatic and terrestrial organisms require low levels of selenium in their diet to sustain
metabolic processes, whereas excess concentrations of selenium that are only an order of
magnitude greater than the required level have been shown to be toxic to fish, apparently due to
generation of reactive oxidized species, resulting in oxidative stress (Palace et.al., 2004). Dietary
requirements in fish have been reported to range from 0.05 to 1.0 mg Se/kg dw (Watanabe et al.
1997). Selenium requirements for optimum growth and liver glutathione peroxidase activity in
channel catfish were reported as 0.25 mg Se/kg dw (Gatlin and Wilson 1984). Estimated
selenium dietary requirements in hybrids of striped bass, based on selenium retention, were
reported as 0.1 mg Se/kg dw (Jaramillo 2006). Selenium deficiency has been found to affect
humans (U.S. EPA 1987), sheep and cattle (U.S. EPA 1987), deer (Oliver et al. 1990), fish
(Thorarinsson et al. 1994; Wang and Lovell 1997; Wilson et al. 1997; U.S. EPA 1987), aquatic
invertebrates (Audas et al. 1995; Caffrey 1989; Cooney et al. 1992; Cowgill 1987; Cowgill and
Milazzo 1989; Elendt 1990; Elendt and Bais 1990; Harrison et al. 1988; Hyne et al. 1993;
Keating and Caffrey 1989; Larsen and Bjerregaard 1995; Lim and Akiyama 1995; Lindstrom
1991; U.S. EPA 1987; Winner 1989; Winner and Whitford 1987), and algae (Doucette et al.
1987; Keller et al. 1987; Price 1987; Price et al. 1987; Thompson and Hosja 1996; U.S. EPA
1987; Wehr and Brown 1985). The predominance of research on selenium deficiency in
invertebrates and algae is related to optimizing the health of test organisms cultured in the
laboratory. A summary of several studies that evaluated the deficiency and/or the sufficiency of
selenium in the diet offish is provided in Appendix E.
2.5 INTERACTIONS WITH MERCURY
The most well-known interactions with selenium occur with both inorganic and organic
mercury, and are generally antagonistic (Micallef and Tyler 1987; Cuvin and Furness 1988;
Paulsson and Lundbergh 1991; Siegel et al. 1991; Southworth et al. 1994; Ralston et al. 2006),
with the most likely mechanism being the formation of metabolically inert mercury selenides
(Ralston et al. 2006; Peterson et al. 2009). However, other studies have found interactions
between mercury and selenium to be additive (Heinz and Hoffman 1998) or synergistic
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(Huckabee and Griffith 1974; Birge et al. 1979). The underlying mechanism for these additive
and synergistic interactions between mercury and selenium are unknown.
2.6 ASSESSMENT ENDPOINTS
Assessment endpoints are defined as "explicit expressions of the actual environmental
value that is to be protected" and are defined by an ecological entity (species, community, or
other entity) and its attribute or characteristics (U.S. EPA 1998). Assessment endpoints may be
identified at any level of organization (e.g., individual, population, community). In the context of
the Clean Water Act, aquatic life criteria for toxic pollutants are typically determined based on
the results of toxicity tests with aquatic organisms in which unacceptable effects on growth,
reproduction, or survival occurred. The goal of criteria is to protect the diversity, productivity,
and stability of aquatic communities. To achieve this goal, the endpoint of criteria assessment is
the survival, growth, and reproduction of a high percentage of species of a diverse assemblage of
freshwater aquatic animals (fish, amphibians, and invertebrates) and plants. Toxicity data are
aggregated into a sensitivity distribution that indicates the impact of the toxicant under study to a
variety of genera representing the broader aquatic community. Criteria are designed to be
protective of the vast majority of aquatic animal species in an aquatic community (i.e.,
approximately 95* percentile of tested aquatic animals representing the aquatic community). As
a result, health of the aquatic community may be considered as an assessment endpoint indicated
by survival, growth, and reproduction.
To assess potential effects on the aquatic ecosystem by a particular stressor, and develop
304(a) aquatic life criteria under the CWA, EPA typically requires the following, as outlined in
the U.S.EPA Ambient Water Quality Criteria Guidelines: acute toxicity test data (mortality,
immobility, loss of equilibrium) for aquatic animals from a minimum of eight diverse taxonomic
groups; as well as chronic toxicity data (e.g., survival, growth and reproduction) for aquatic
animals from 8 taxonomic groups (described in more detail below). The diversity of tested
species is intended to ensure protection of various components of an aquatic ecosystem. In the
case of bioaccumulative compounds like selenium, these acute toxicity studies do not address
risks that result from exposure to chemicals via the diet (through the food web). They also do not
account for the slow accumulation kinetics of many bioaccumulative compounds such as
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selenium and may underestimate effects from long-term accumulation in different types of
aquatic systems (SAB 2005).
Because the most sensitive adverse effects of selenium are reproductive effects on the
offspring of exposed fish, chronic effects are the focus of this selenium assessment. Shorter-term
intermittent or pulsed exposures to selenium may result in bioaccumulation through the aquatic
food web and may subsequently adversely affect fish reproduction, and such measures of effect
are therefore estimated from chronic assessment endpoints.
Chronic toxicity test data (longer-term survival, growth, or reproduction) for aquatic
animals are needed from a minimum of eight diverse taxonomic groups (or less generically,
[minimum of three taxa] if the derivation is based on an acute to chronic ratio). The diversity of
tested species is intended to ensure protection of various components of an aquatic ecosystem.
Specific minimum data recommendations or requirements (MDRs) identified for development of
criteria in the EPA Ambient Water Quality Criteria Guidelines require aquatic animal toxicity
data from:
1. the family Salmonidae in the class Osteichthyes ,
2. a second family in the class Osteichthyes, preferably a commercially or
recreationally important warmwater species (e.g., bluegill, channel catfish, etc.),
3. a third family in the phylum Chordata (may be in the class Osteichthyes or may
be an amphibian, etc.),
4. a planktonic crustacean (e.g., cladoceran, copepod, etc.),
5. a benthic crustacean (e.g., ostracod, isopod, amphipod, crayfish, etc.),
6. an insect (e.g., mayfly, dragonfly, damselfly, stonefly, caddisfly, mosquito,
midge, etc.),
7. a family in a phylum other than Arthropoda or Chordata (e.g., Rotifera,
Annelida, Mollusca, etc.), and
8. a family in any order of insect or any phylum not already represented.
Acceptable quantitative chronic values for selenium are available for six of the eight
MDRs (requirements 1, 2, 3, 6, 7, and 8). Acceptable chronic values for selenium are not
available for two of the MDRs (requirements 4 and 5: planktonic and benthic crustaceans,
respectively). Following the approach of U.S. EPA (2008b), which was reviewed by the Science
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Advisory Board, if information is available to demonstrate that an MDR is not sensitive, then a
surrogate value can be used in place of actual toxicity data to represent the missing MDR. Based
on the data estimating the sensitivity of insects (Centroptilum triangulifer), rotifers (Brachionus
calyciflorus), and oligochaetes (Lumbriculus variegatus), EPA determined that invertebrates
(e.g. insects and crustaceans) are less sensitive to selenium than fish. Therefore, the available
fish data were used in the genus-level sensitivity distribution to derive the chronic selenium
criterion (Note: invertebrate data were included in the sensitivity distribution for the whole body
criterion element to demonstrate that the derivation of the criterion element based on the fish
egg-ovary to whole body translated values protected invertebrates based on the sensitivity range
of the available species).
The U.S.EPA Ambient Water Quality Criteria Guidelines also require at least one
acceptable test with a freshwater alga or vascular plant. If plants are among the aquatic
organisms most sensitive to the stressor, results of a plant in another phylum should also be
available. A relatively large number of tests from acceptable studies of aquatic plants were
available for possible derivation of a Final Plant Value. However, the relative sensitivity of fresh
and saltwater plants to selenium (Appendix F) is less than fish so plant criteria were not
developed.
The available scientific evidence indicates that for selenium, critical assessment
endpoints for aquatic species are offspring mortality and severe development abnormalities that
affect the ability offish to swim, feed and successfully avoid predation, resulting in impaired
recruitment of individuals into fish populations. Selenium enrichment of reservoir environments
(e.g., Belews Lake, NC (Lemly 1985), Hyco Reservoir (DeForest 1999), and Kesterson
Reservoir, CA (Ohlendorf 1986) are well documented and demonstrate that adverse effects
resulted from bioaccumulative processes at different levels of biological organization, resulting
in population-level reductions of resident species.
2.7 MEASURES OF EFFECT
Each assessment endpoint requires one or more "measures of ecological effect", which
are defined as changes in the attributes of an assessment endpoint itself or changes in a surrogate
entity or attribute in response to chemical exposure. Ecological effects data are used as measures
of direct and indirect effects to growth, reproduction, and survival of aquatic organisms.
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The toxicity testing data available for any given pollutant vary significantly, depending
primarily on whether any major environmental issues are raised. An in-depth evaluation of
available data for selenium has been performed by EPA to determine data acceptability and
quality, based on criteria established in the U.S.EPA Ambient Water Quality Criteria Guidelines.
In traditional chronic tests used in many EPA aquatic life criteria documents, organisms
are exposed to contaminated water but fed a diet grown in uncontaminated media not spiked with
the toxicant prior to introduction into the exposure chambers. Such tests are not suitable for
deriving a criterion for a bioaccumulative pollutant unless (1) effects are linked to concentrations
measured in appropriate tissues, and (2) the route of exposure does not affect the potency of
residues in tissue. For selenium, the first condition might be met, but the second condition is not,
because the route of selenium exposure appears to influence the potency of a given tissue residue
(Cleveland et al. 1993; Gissel-Nielsen and Gissel-Nielsen 1978). Consequently, toxicity tests
with water-only exposures (and any tests not relying on dietary exposure) are not included in this
assessment.
Selenium toxicity is primarily manifested as reproductive impairment due to maternal
transfer, resulting in embryo mortality and teratogenicity. Measurements of selenium in fish
tissue are most closely linked to the chronic adverse effects of selenium (Chapman et al. 2010),
since chronic selenium toxicity is based on the food-chain bioaccumulation route, not a direct
waterborne route. In this selenium criterion document, water column criterion element
concentrations for selenium were derived from fish tissue concentrations by modeling selenium
transfer through the food web. The next sections describe approaches used to establish selenium
effects concentrations in fish tissue and to relate the concentrations in fish tissue to
concentrations in water.
2.7.1 Fish Tissue
Chronic measures of effect concentrations are the ECio, EC20, No Observed Effect
Concentration (NOEC), Lowest Observed Effect Concentration (LOEC), and Maximum
Acceptable Toxicant Concentration (MATC). The ECio is the concentration of a chemical that is
estimated to result in a 10 percent effect in a measured chronic endpoint (e.g., growth,
reproduction, or survival); the EC20 corresponds to 20 percent effect. The NOEC is the highest
chemical concentration at which none of the observed effects are statistically different from the
control, as determined by hypothesis testing. The LOEC is the lowest test concentration at which
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observed effects are found to be statistically different from the control. The MATC is the
geometric mean of the NOEC and LOEC.
Whenever possible, estimates of selenium concentrations associated with a low level of
effect (i.e., ECio) were calculated for each study using the computer program TRAP (v.1.22),
Toxicity Relationship Analysis Program (U.S. EPA 2013). The program is based on a regression
approach that models the level of adverse effects as a function of increasing concentrations of the
toxic substance. With the fitted model it is possible to estimate the contaminant concentration
associated with a small effect. Adverse effects were modeled as a sigmoid function of the
logarithmic concentrations of the toxic substance and concentration-response data were analyzed
using TRAP software (U.S. EPA 2013). When individual response level data were available,
tolerance distribution analysis was performed. When only treatment or replicate level data were
available, nonlinear regression was performed.
Only studies with a reference site (field surveys) or control treatment (experimental
studies) were included in the analysis, because response levels at these low (background)
selenium concentrations were the most influential points for calculating the estimated response
level at a selenium concentration of zero (yo).
When considering the use of the ECio versus the EC20, an ECio was determined to be a
more appropriate endpoint for tissue-based criteria given the nature of exposure and effects for
this bioaccumulative chemical. EC2os have historically been used in the derivation of EPA
criteria applicable to the water medium. While water concentrations may vary rapidly over time,
tissue concentrations of bioaccumulative chemicals are expected to vary gradually. Thus, where
concentrations of selenium in fish tissue are used as an effect threshold, there is potential for
sustained impacts on aquatic systems, relative to chemicals that are not as bioaccumulative. This
calls for use of a lower level of effect to attain sufficient protection. Further, the ECio was also
preferred over the NOEC or LOEC as these measures of effect are influenced by study design,
specifically the concentrations tested, the number of concentrations tested, the number of
replicates for each concentration, and the number of organisms in each replicate. As noted by
Campbell (2011), ECios and NOECs are generally of similar magnitude, but ECios have the
advantage of being more reproducible than NOECs (Van der Hoeven et al. 1997; Warne and van
Dam 2008). NOECs and MATCs are generally presented if calculated by the original
investigators, but were not used where an ECio could be calculated. The four lowest egg-ovary
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Genus Mean Chronic Values (GMCVs), whose exact values influence the calculation of the egg-
ovary criterion, are all based solely on ECios. NOECs contribute to some of the GMCVs for less
sensitive species.
In this document, chronic values are presented as tissue concentrations of total selenium
in units of mg/kg dry weight (dw). Studies of chronic toxicity of selenium to aquatic organisms
measure concentrations in distinct tissues (e.g., whole body, ovaries, eggs, muscle, and liver) and
report these values as either wet weight (ww) or dw. Studies reporting tissue concentrations only
based on wet weight were converted to dry weight using tissue-specific and species-specific
conversion factors. When wet to dry weight conversion factors were not available for a given
species, conversion factors for a closely related taxon were used. In deriving the egg or ovary
tissue criterion, chronic values are for those tissues directly measured in the study. Tissue-to-
tissue conversions (e.g., to estimate concentrations in an unmeasured tissue from a study's
measured tissue) involve some uncertainty because of variability in tissue concentration ratios
(deBruyn et al. 2008; Osmudson et al. 2007). Tissue-to-tissue conversions were needed for
calculating the reproductive toxicity-based whole-body and muscle chronic criterion element and
water criterion concentration elements.
The overall assessment evaluates both reproductive and non-reproductive studies.
Selenium concentrations measured directly in eggs or ovaries from reproductive (maternal
transfer) studies are used to derive the egg/ovary criterion element, and corresponding selenium
concentrations in whole body or muscle tissue resulting in reproductive effects are estimated
using conversion factors. Direct measurements of selenium concentrations in whole-body or
muscle from non-reproductive studies are used to examine non-reproductive, chronic effects,
such as impairments to growth.
2.7.2 Water
While state monitoring programs may sample ambient waters for selenium, widespread
measurements of selenium in fish tissue are relatively rare. Therefore, EPA is estimating chronic
measures of effect in the water column. The chronic criterion element for the water column is the
30-day average concentration that corresponds to the concentration of selenium in fish tissue
estimated to result in a 10 percent effect in fish for a specific water body type (lotic or lentic
water bodies as described below in Section 3.2.4). The chronic criterion element for the water
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column is derived by modeling trophic transfer of selenium through the food web resulting in the
fish tissue concentration that yields the chronic reproductive effects of concern.
The EPA collaborated with the United States Geological Survey (USGS) to develop a
model (later published in Presser and Luoma, 2010) that relates the concentration of selenium in
fish tissue to the water column. The approach is based on bioaccumulation and trophic transfer
through aquatic system food-webs. Model parameters are calculated using both field and
laboratory measurements of selenium in water, particulate material (algae, detritus and
sediment), invertebrates, fish whole-body, and fish egg-ovary. This model (which is a set of
equations) is described in more detail in Section 3.2.1.
-^^ -^^
2.7.3 Summary of Assessment Endpoints and Measures of Effect
The typical assessment endpoints for aquatic life criteria are based on effects on growth,
reproduction, or survival of the assessed taxa. These measures of effect on toxicological
endpoints of consequence to populations are provided by results from toxicity tests with aquatic
plants and animals. The toxicity values (i.e., measures of effect expressed as genus means) are
used in the genus sensitivity distribution of the aquatic community to derive the aquatic life
criteria. Endpoints used in this assessment are listed in Table 2.2.
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Table 2.2. Summary of Assessment Endpoints and Measures of Effect Used in Criteria
Derivation for Selenium.
Assessment Endpoints for the Aquatic
Community
Measures of Effect
Survival, growth, and reproduction of
freshwater fish, other freshwater vertebrates,
and invertebrates
For effects from chronic exposure:
1. EC 10 concentrations in egg and ovary, for
offspring mortality and deformity.
2. Estimated reproductive ECio in whole
body and muscle.
3. Estimated concentrations (|ig/L) in water
linked to egg-ovary ECios by food web-
modeling.
4. Intermittent water concentrations yielding
exposure equivalent to the above.
For acutely lethal effects:
Acute toxicity effects based on standard
water column-only toxicity testing are not
provided here for selenium, due to the
dominant significance of chronic effects.
Note: Chronic criterion is expected to be
protective of acute effects.
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2.7.4 Conceptual Model of Selenium Effects on Aquatic Life
Selenium Sources
Naturally elevated selenium in soils - agricultural irrigation practices (Western US only)
Mining activities - coal, metals and sulfide minerals, phosphate
Selenium in Water Column
Algal/Plant Transformation and Enrichment:
• As function of sorption to particulates (sediment, algae, detritus)
As function of system hydrodynamics, lotic & lentic systems, residence time
Initial Trophic Transfer
from phytoplankton, periphyton, macrophytes, detritus, & sediment
Secondary Trophic Transfer from
macroinvertebrates/
icthyoplankton/ other zooplankton
Tertiary Trophic Transfer from
lower trophic level fish
To higher trophic level fish
I Reproductive Impairment.
Larval skeletal deformities.
Larval mortality.
Population decline
Figure 2.3. Diagram of selenium partitioning, bioaccumulation, and effects in the aquatic
environment.
The conceptual model links sources, transformation and uptake through media phases,
and consumer transfer and dynamics reflective of the movement of selenium through ecosystems
(Figure 2.3). Diet is the dominant pathway of selenium exposure for both invertebrates and
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vertebrates. Selenium moves from water to particulates, a collection of biotic and abiotic
compartments that includes primary producers, detritus, and sediments, which form the base of
aquatic food webs. Transfer from particulates to primary consumers (e.g., macroinvertebrates) to
fish is species specific. Knowledge of the food web is one of the keys to determining which
biological species or other ecological characteristics will be affected.
During the development of CWA section 304(a) criteria, EPA assembles all available test
data and considers all the relevant data that meet acceptable data quality and test acceptability
standards. This criterion update document is specific to selenium in fresh water. Chronic
criterion elements for selenium are protective concentrations measured in fish tissue and related
to protective water concentrations generated using food-web modeling. Further modeling is used
to estimate short-term concentrations in water from intermittent or pulsed exposures that are
protective against the chronic effect.
2.7.5 Analysis Plan for Derivation of the Chronic Fish Tissue-Based Criterion Elements
Data for possible inclusion in the selenium dataset were obtained primarily by search of
published literature using EPA's public ECOTOX database (up to July 2013). These studies were
screened for data quality as described in the U.S.EPA Ambient Water Quality Criteria
Guidelines, and adjusted for factors related to dietary lab or field exposure, which were not
considered at the time the Guidelines were written. Additional data were considered and
reviewed for inclusion in this criterion based on the public and peer review comments on the
2014 "External Peer Review Draft" criterion document.
Chronic toxicity studies (both laboratory and field studies) were further screened to
ensure they contained the relevant chronic exposure conditions of selenium to aquatic organisms
(i.e., dietary, or dietary and waterborne selenium exposure), measurement of chronic effects, and
measurement of selenium in tissue(s). The criterion derivation uses only those studies in which
test organisms were exposed to selenium in their diet, because such studies most closely replicate
real-world exposures (diet and/or diet plus water). This approach accords with findings and
recommendations of the 2009 SETAC Pellston Workshop (Chapman et al. 2009, 2010).
EPA grouped studies based on whether the effects were chronic reproductive (e.g.,
effects on offspring survival or morphology) or chronic non-reproductive (e.g., juvenile growth
and survival). At the 2009 Pellston workshop (Chapman et al. 2009, 2010), a group of 46 experts
in the area of ecological assessment of selenium in the aquatic environment agreed that the most
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important toxicological effects of selenium in fish arise following maternal transfer of selenium
to eggs during vitellogenesis, resulting in selenium exposure when hatched larvae undergo yolk
absorption. Such effects include larval mortality or permanent developmental malformations,
such as skeletal and craniofacial deformities. Therefore, the chronic fish-tissue-based criterion
elements are based on reproductive effects only.
The egg-ovary Species Mean Chronic Values (SMCVs) were calculated from the chronic
values (ECios and occasionally NOECs) obtained from the relevant toxicity tests. Genus Mean
Chronic Values (GMCVs) were calculated from the SMCVs and then rank-ordered from least to
most sensitive. The egg-ovary Final Chronic Value (FCV) was calculated from regression
analysis of the four most sensitive GMCVs, in this case extrapolating to the 5* percentile of the
distribution represented by the tested genera. The FCV directly serves as the fish tissue egg-
ovary criterion concentration element without further adjustment because the underlying ECios
represent a low level of effect (per the U.S.EPA Ambient Water Quality Criteria Guidelines).
For the whole-body and muscle criterion concentrations, the egg-ovary GMCVs were
converted to estimated equivalent whole-body or muscle GMCVs. The criterion concentration
element expressed as whole-body or as muscle concentration was calculated in a manner similar
to the egg-ovary criterion element using conversion factors described below, from their
respective genus-level sensitivity distributions.
2.7.6 Analysis Plan for Derivation of the Fish Tissue Criterion Elements Duration
A numerical value for the fish tissue criterion elements averaging period, or duration, is
specified as instantaneous, because fish tissue data provide point, or instantaneous,
measurements that reflect integrative accumulation of selenium over time and space in the fish at
a given site. Selenium concentrations in fish tissue are generally expected to change only
gradually over time (Section 3.2.6 and Appendix J) in response to environmental fluctuations;
thus, there would be relatively little difference in tissue concentrations with different averaging
period durations.
2.7.7 Analysis Plan for Derivation of Chronic Water-based Criterion Element
The relationship between the ambient concentration of selenium in water and the
concentration of selenium in the eggs or ovaries offish is primarily through trophic transfer of
selenium, which is greatly affected by site-specific conditions. EPA used a peer-reviewed model
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to derive water concentrations from the egg-ovary criterion that explicitly recognizes partitioning
of selenium in water and particulate material (algae, detritus, and sediment), and trophic transfer
from particulate material to aquatic invertebrates, from invertebrates to fish, and partitioning in
fish whole-body and fish eggs and ovaries. The method is composed of five main steps:
1. Formulate a mathematical equation relating the concentration of selenium in the eggs and
ovaries offish to the ambient concentration of selenium in the water column.
2. Develop parameters needed to use the mathematical equation formulated in step 1 from
available empirical or laboratory data related to selenium bioaccumulation in aquatic systems
and aquatic organisms.
3. Classify categories of aquatic systems where a single water column concentration would be
adequately protective by evaluating the bioaccumulation potential at the base of the aquatic
food web.
S^
4. Translate the egg-ovary criterion element to an equivalent water column concentration at
each aquatic site.
5. Apply a statistical threshold to the distribution of translated water column concentrations for
each aquatic system category to yield a water column concentration value that would be
protective of each aquatic system category.
EPA worked with the United States Geological Survey to derive a translation equation to
estimate the site-specific concentration of selenium in the water column corresponding to the
egg-ovary criterion concentration. This equation utilizes a mechanistic model of bioaccumulation
previously published in peer-reviewed scientific literature (Luoma et. al. 1992; Wang et. al.
1996; Luoma and Fisher 1997; Wang 2001; Schlekat et al. 2002b; Luoma and Rainbow 2005;
Presser and Luoma 2006; Presser and Luoma 2010; Presser 2013). The equation uses site-
specific food web models, species-specific Trophic Transfer Factor (TTF) values, egg-ovary to
whole-body conversion factor (CF) values, and a site-specific enrichment factor (EF) values to
calculate a site-specific water column concentration element from the egg-ovary criterion
element.
Empirical or laboratory data related to selenium bioaccumulation in aquatic organisms
are needed to calculate species-specific TTF and CF parameters and a site-specific EF parameter.
EPA obtained these data by utilizing their extensive selenium library of published papers and
reports, and by searching published literature using EPA's public ECOTOX database and other
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publically available data received through solicitation of public comments on the 2014 "External
Peer Review" draft. Studies were screened using the same data quality guidelines as described
above. Relevant studies contained selenium measurements from field studies (water, particulate
material, and aquatic organisms) or contained laboratory data on physiological parameters of
selenium bioaccumulation in aquatic organisms. Literature searches for information on selenium
associated with particulate matter included searches for data on all forms of algae, detritus,
inorganic suspended material, and sediment.
EPA compiled a collection of selenium concentration measurements from acceptable
field studies. Measurements were accepted if the study indicated the samples were collected in
the field, and the study identified the unit of measure, the media from which the measurement
was made, the location from where the sample was taken, and the date the sample was collected.
EPA only used data from studies with adequately described field collection protocols and where
concentrations were within the bounds of concentrations found using modern, rigorous protocols
in similar systems (Safiudo-Wilhelmy et al. 2004). The spatial precision of field data sample
collection locations were generally at the site level, although aggregate measurements were also
included if exposure conditions were considered similar (e.g., averages of single or composite
measurements from several locations in the same aquatic system). The temporal precision of
sample collection times were usually at the level of the day they were collected, although some
studies only provided enough information to determine the week, month, or year. If the day a
series of samples were collected was not reported but the study provided information that
indicated the samples were taken concurrently, EPA noted sample precision, but assigned a
single effective collection date to all the samples.
EPA also compiled a collection of physiological coefficients for food ingestion rate (IR),
selenium assimilation efficiency (AE), and rate of selenium loss (ke) from published literature.
Coefficients were accepted if the studies provided either the actual measurements or sufficient
information to derive them, and were reported in standard units (ke. /d; AE: %; IR: g/g-d) or
could be converted to standard units. Even though IR can be highly variable (Whitledge and
Hayward 2000), IR values of surrogate species were occasionally used.
EPA accounted for bioaccumulation variability across aquatic sites by evaluating the
parameter EF (representing the partitioning of selenium between the dissolved and particulate
state) from representative aquatic systems. The parameter EF is a measure of bioaccumulation
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potential because it quantifies the transfer of selenium from the water column to particulate
material, which is the single most influential step in selenium bioaccumulation (Chapman et al.
2010). EPA calculated EF values for a set of aquatic systems using data from published literature
and applied statistical methods to distinguish categories with similar bioaccumulation
characteristics. On this basis, a single water column concentration is deemed adequately
protective when it is derived using data from aquatic sites in the same category. EPA translated
the egg-ovary criterion element into a set of water concentration values and derived a water
column criterion element for each aquatic system category using a percentile of the water column
concentrations for each category. To ensure adequate protection, EPA selected the 20* percentile
of the distribution of median water column values as the statistical cut-off (see Section 3.2.5).
Figure 2.4 diagrams the conceptual framework EPA used to derive water column criterion
element values from the egg-ovary criterion element.
2.7.8 Analysis Plan for Derivation of the Water Criterion Elements Duration
A numerical value for the lentic and lotic water criterion elements averaging period, or
duration, is specified as a 30 day average, because the presence of selenium in water is the initial
step in the process of bioaccumulation from the water column to fish tissue. The
bioaccumulation process for selenium takes place over a longer term than typically observed for
acute and chronic effects on aquatic life based on water concentrations. Therefore,
concentrations of selenium in lentic and lotic waters must remain elevated for a sufficient length
of time to provide a source of selenium in the water column leading to elevated fish tissue levels
that could impact fish reproduction. The derivation of a protective water averaging period from
kinetic modeling considerations is described in Section 3.2.6 and in Appendix J.
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Egg-ovary criterion element I
Representative aquatic systems I
Egg/ovary - whole body Conversion Factors (CF)
Concentrations in whole-body offish I
Trophic Transfer Functions (77?)
Concentrations in prey
Trophic Transfer Functions (77?)
Concentrations in particulate material
Site-specific Enrichment Factors (EF)
Distribution of water concentrations
Statistical threshold
Water criterion element
Figure 2.4. Conceptual model for translating the selenium egg-ovary concentration to a
water column concentration.
2.7.9 Analysis Plan for Intermittent-Exposure Water-based Criterion Element Derivation
Like the chronic water criterion element, the intermittent-exposure criterion element
protects against cumulative exposure of selenium from multiple short-term discharges that may
cause an excursion of the fish tissue criterion element. EPA derived the intermittent exposure
criterion element directly from the chronic water criterion element by algebraically rearranging
the chronic water criterion element to establish a limit on an intermittent elevated concentration
occurring over a specified percentage of time, while simultaneously accounting for natural or
anthropogenic background concentrations (see Section 3.3).
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3 EFFECTS ANALYSIS FOR FRESHWATER AQUATIC ORGANISMS
3.1 CHRONIC TISSUE-BASED SELENIUM CRITERION ELEMENT CONCENTRATION
Data were obtained primarily by search of published literature using EPA's public
ECOTOX database. The most recent ECOTOX database search extended to July 2013, but this
draft also reflects data either gathered or received by the Agency up to December 2014 based on
information from the public comment period and external expert peer review provided in
response to the "External Peer Review Draft" published in May 2014. All available, relevant,
and reliable chronic toxicity values were incorporated into the appropriate selenium AWQC
tables and used to recalculate the FCV, as outlined in detail in the U.S.EPA Ambient Water
Quality Criteria Guidelines.
The chronic values determined from acceptable chronic toxicity studies were separated
into reproductive endpoint and non-reproductive endpoint categories. Although both sets of
endpoints assess effects due to selenium on embryo/larval or juvenile development and survival
and growth, the fundamental difference is exposure route (inherent in test design). That is, the
fundamental difference is whether the aquatic organisms (e.g., fish) were directly exposed to
selenium in the diet and water column or exposed via maternal transfer of selenium to the
eggs/ovaries prior to reproduction. In studies with reproductive endpoints, parental females are
exposed to selenium and the contaminant is transferred from the female to her eggs. In the
selenium-exposed females, selenium replaces sulfur in vitellogenin, the primary yolk precursor,
which is transported to the ovary and incorporated into the developing ovarian follicle (Janz et al.
2010). In most but not all of these studies, progeny from these females were not additionally
exposed to aqueous selenium. The chronic values derived for the reproductive effects (survival,
deformities, and edema) are based on the concentration of selenium in the eggs or ovary, the
tissues most directly associated with the observed effects. In contrast, in studies grouped under
non-reproductive effects (usually larval and/or juvenile survival or growth), the tested fish had
no maternal pre-exposure to selenium. Chronic values for non-reproductive effects are based on
the concentration of selenium in tissues measured in the study: muscle, liver and/or whole body.
The reproductive endpoint studies applied to the derivation of the chronic criterion
elements are described below. Less definitive reproductive studies that are not directly applied to
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the criterion derivation are described in Section 6.1.2 and in Appendix C. Nonreproductive
studies are described in Section 6.1.9.
3.1.1 Acceptable Studies of Fish Reproductive Effects of Four Most Sensitive Genera
Below is a brief synopsis of the experimental design, test duration, relevant test
endpoints, and other critical information regarding the four sensitive genera that drive the
calculation of each specific chronic value. The studies in this section involve effects on the
offspring of exposed female fish. Data are summarized in Table 3.1. Details of these studies and
other chronic studies considered for criteria derivations are contained in Appendix C.
3.1.1.1 Acipenseridae
3.1.1.1.1 Acipemer trammontanus (white sturgeon)
Linville (2006) evaluated the effect of elevated dietary selenium on the health and
reproduction of white sturgeon. Adult female white sturgeon (approximately 5 years old) were
fed either a control diet (no added selenium, 1.4 mg/kg Se) or a diet spiked with selenized yeast
(34 mg/kg Se) for six months in a freshwater flow through system. At the end of the dietary
exposure, females were induced to spawn and fertilized with non-exposed male milt. Eggs were
hatched in jars, keeping eggs from each female separate. Progeny from two control females and
three treatment females were examined for length, weight, edema and deformities. No selenium
effects were observed for length or weight of larvae but effects were observed for both edema
and skeletal deformities. Selenium concentrations in eggs from the control fish were 1.61 and
2.68 mg/kg dw, and were 7.61, 11 and 20.5 mg/kg dw in eggs from the treatment fish. The larvae
hatched from these eggs had respectively 0, 0, 0, 13.3, and 27.8% occurrence of the combined
effects of edema and deformities.
An ECio of 16.3 mg/kg dw for total larval deformities (edema + skeletal) in response to
selenium concentrations in eggs was calculated using the threshold sigmoid nonlinear regression
model in TRAP (v.1.22). Because the ECio is based only on the partial response (27.8% effect) at
the highest egg selenium concentration (20.5 mg/kg Se egg), the modeled ECio was sensitive to
the slope of the model, and the ECio of 16.3 mg/kg was the most conservative model across a
range of slopes with identical goodness of fit (see Appendix C for details). An ECio based on
only one partial response would not ordinarily be included in the chronic data set, but there are
supporting data that suggest the federally-listed threatened species green sturgeon is also
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sensitive to selenium (see Section 6.3 Protection of Threatened or Endangered Species). The
white sturgeon ECio of 16.3 mg/kg egg dw is included in the data set because there are data
indicating reproductive effects at this concentration. This species, which is listed as endangered
in specific locations, such as the Kootenai River white sturgeon in Idaho and Montana. The
white sturgeon is also a taxonomic surrogate for other freshwater sturgeon species (e.g.,
shovelnose sturgeon) that are threatened or endangered.
3.1.1.2 Salmonidae
Acceptable studies were available for three salmonid genera, Oncorhynchus, Salvelinus
and Salmo. All of these studies evaluated the effects of selenium on salmonid embryo/larval
survival and deformity and used wild-caught adults taken from selenium contaminated streams
and spawned for effects determination. Exposure for all studies was therefore through the
parents. Summaries of the studies with Oncorhynchus and Salvelinus malma are discussed in
Section 6.1.2.3; brown trout (Salmo trutta) is discussed below.
3.1.1.2.1 Salmo trutta (brown trout)
Formation Environmental (2011) collected adult female and male brown trout from sites
with low and high selenium exposure in the vicinity of a phosphate mine located in Southeastern
Idaho in November 2007. Eggs were collected from 26 gravid females across three sampling
locations, fertilized with milt collected from several males from the same site and taken to the
laboratory for hatching and observation of larval malformations and survival. In addition to the
field collected fish, fertilized eggs of twelve females from two separate hatcheries were used in
the study. The study had two phases, hatch-to-swim up, and swim up-to-15 days post swim-up.
There are two experimental complications that affect the interpretation of these data: (a) elevated
deformity rates among the offspring that were to serve as hatchery-originated method controls
(very low selenium exposure) and among some of the low exposure field-collected organisms,
and (b) the accidental loss of a number of individuals from several treatments during the 15-day
post swim up portion of the test due to overflow of the tank water. This document's analysis of
the revised counts from AECOM (2012) builds upon and supersedes EPA's 2012 analysis
(Taulbee et al. 2012), peer reviewed by ERG (2012).
A total of seven ECios were calculated for this study. Six of the ECios were calculated for
the full test, in which the hatch-to-swim up and the swim up-to-15 days post swim up phases of
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the test were combined. For the full hatch through 15-days post swim up test, three separate
endpoints were measured: larval survival, larval deformities (% normal), and combined larval
survival and deformities (% normal survivors). For each of these three endpoints, two ECios
were calculated under two scenarios regarding the assumed health of the larvae lost during the
overflow of several tanks during the second phase of the test. In the worst case scenario, all
larvae lost during the overflow accident were assumed to have been dead or deformed. In the
optimistic scenario, all larvae lost during the overflow accident were removed from the ECio
calculations, under the assumption that they had the same rates of mortality and deformity as
those not lost. All ECio calculations included both field and hatchery treatments. The range of
the six ECios calculated for the full hatch through 15-days post swim up test was 16.78 - 21.94
mg/kg egg dw. Larval survival was the most sensitive endpoint, followed by the combined
survival and deformities endpoint, with deformities being the least sensitive. For each of the
three endpoints, the ECio for the worst case scenario was lower than the ECio for the optimistic
scenario. Because of uncertainties as to how to best address the loss offish during the overflow
event during the second phase of the test, an ECio for survival during only the first portion of the
test was calculated. The ECio for this endpoint was 18.09 mg/kg egg dw for larval survival
during the first portion of the test, hatch to swim up. Section 6.1.6 provides a summary of the
analysis that led to the final selection of the ECio for larval survival during the first portion of the
test. Appendix C presents details of the study and analysis.
3.1.1.3 Centrarchidae
3.1.1.3.1 Lepomis macrochirus (bluegill sunfish)
In a laboratory study, Doroshov et al. (1992) exposed adult bluegill for 140 days to three
dietary concentrations of seleno-L-methionine added to trout chow. Near the end of the
exposure, ripe females were induced to ovulate and ova were fertilized in vitro with milt stripped
from males. Fertilized eggs were sampled for fertilization success and selenium content. They
were also used in two tests, (a) a larval development study during the first 5 days after hatching,
and (b) a 30-day embryo-larval test. In the 5-day larval test, the average proportion of larvae with
edema was 0% at an egg concentration of 8.33 mg Se/kg (the first treatment), 5% at an egg
concentration of 19.46 mg Se/kg dw (the second treatment), and 95% at an egg concentration of
38.39 mg Se/kg dw (the highest treatment). The latter two were statistically different from the
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control (0% edema). All edematous larvae died in the high treatment. In the 30-day larval
survival test, statistical difference from the control was only found in the highest test treatment
for survival and growth (length and weight) measurements. The ECio calculated with TRAP for
the incidence of edema in the 5-day larval bioassay is 20.75 mg Se/kg dw in eggs.
A similar study with similar results was done by Coyle et al. (1993) in which two year
old pond-reared bluegill sunfish were exposed in the laboratory and fed (twice daily ad libitum)
Oregon moist™ pellets containing increasing concentrations of seleno-L-methionine. Water
concentrations were nominal 10 jag Se/L. The fish were grown under these test conditions for
140 days. Spawning frequency, fecundity, and percentage hatch were monitored after 60 days
when spawning began to occur. There was no effect from the highest dietary selenium
concentration (33.3 mg Se/kg dw) on adult growth, condition factor, gonadal somatic index, or
the various reproductive endpoints (Appendix C). The survival of newly hatched larvae,
however, was markedly reduced; only about 7 percent of larvae survived to 5 days post-hatch in
the high dietary treatment. The TRAP-calculated ECio value for larval survival is 24.55 mg
Se/kg dw in eggs.
Hermanutz et al. (1992), and Hermanutz et al. (1996) exposed bluegill sunfish to sodium
selenite spiked into artificial streams (nominal test concentrations: 0, 2.5, 10, and 30 jag Se/L)
which entered the food web, thus providing a simulated field exposure (waterborne and dietary
selenium exposure). In an effort originally intended to improve the rigor of the statistical analysis
of the Hermanutz et al. (1996) data, Tao et al. (1999) re-examined the raw data records and made
corrections to the counts. This criterion document considers the Hermanutz et al. (1992) data and
the Tao et al. (1999) re-examination of Hermanutz et al. (1996).
These data come from a series of three studies lasting from 8 to 11 months, conducted
over a 3-year period. All three studies began with exposure of adult bluegill sunfish in the fall,
and with respective studies ending in the summer of the following year. Temperatures averaged
4.6, 4.1 and 4.5°C during the winter months and averaged 26.4, 23.9 and 22.4°C during the
spawning months (June-July) for Studies I, II and III, respectively. Spawning activity was
monitored in the stream, and embryo and larval observations were made in situ and from
fertilized eggs taken from the streams and incubated within egg cups in the laboratory. None of
the adult bluegill exposed to the highest concentration of selenium in the water (Study I, mean
measured concentration equal to 29.4 ng/L) survived the entire exposure period (although a few
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did survive to spawn). Embryo-larval effects were observed in the selenium-dosed streams in
both Study I and Study II. The incidence of edema, lordosis, hemorrhage and larval survival in
the one stream concentration common to both Study I and II, 10 ug/L, ranged from 80 to 100
percent, 5 to 18 percent, 27 to 56 percent, and 29 to 58 percent, respectively over the three years
(combined egg cup and nest observations). Edema, lordosis, and hemorrhage in the lowest
stream concentration in Study II, 2.5 ug/L, ranged from 0 to 4 percent, 0 to 25 percent, and 3.6 to
75 percent, respectively (combined egg cup and nest observations); larval survival was 71.6
percent (72 and 75 percent in the control streams). See Hermanutz 1996 and 1992 in Appendix C
for more detail. The effects were not observed in larvae from fish that were not exposed to
elevated concentrations of selenium (control treatment). The mean concentrations of selenium in
bluegill ovaries ranged from 0.8 to 2.5 mg/kg dw in the control, 7.6 to 10.9 mg/kg dw in the 2.5
ug/L treatment, and 17.7 to 30.0 mg/kg dw in the 10 jag Se/L treatment (values represent both
Study I and II for the control and 10 ug/L treatments).
The embryo-larval data for continually exposed streams of Studies I and II of this
experiment were combined and analyzed in response to measured selenium concentration in the
maternal ovaries (mg/kg dw) using TRAP. In the recovering streams of Study II and Study III no
effects were observed at concentrations that had previously caused effects in the continually
exposed streams. Recovering streams were therefore excluded from this criterion document's
analysis because they do not reflect the type of system to which water quality criteria are most
commonly applied (i.e., systems receiving existing waterborne pollutant discharges). That is,
Study III consisted of the addition of new adult bluegill to the same streams that had received the
2.5, 10, and 30 ug/L sodium selenite during previous studies, but with all continued external
dosing of selenite halted. The adult bluegills exposed only to dietary selenium present in the
Study III food web accumulated selenium to levels very near to those accumulated during Study
II in which aqueous selenium was also present, demonstrating the importance of diet on selenium
accumulation. There were no effects (no effect on larval survival, 0 percent deformities, 0
percent hemorrhaging), on the bluegill progeny in Study III even from fish that accumulated
26.04 and 10.17 mg/kg dw in the recovering 10 ug/L streams, and 15.92 mg/kg dw in the
recovering 30 ug/L stream. The absence of effects at high tissue levels in the recovering streams
of Study III provides experimental corroboration for the field observations of biological recovery
in Belews Lake and Hyco Reservoir after selenium loads were reduced, but while tissue
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concentrations remained relatively high (Lemly 1997'a; Crutchfield 2000; Finley and Garrett
2007). Because Study III involved new (naive) fish added to previously contaminated streams,
neither acclimation nor adaptation would seem to explain this phenomenon. Overall, the
implication is that for some period of time, recovering systems might exceed tissue criterion
concentrations even though the effects of selenium have been mitigated.
Several endpoints were analyzed by TRAP independently (% edema, % lordosis, and %
hemorrhage) and in combination (% edema and % larval survival) relative to ovary selenium
concentration. The best fit and most sensitive was the combined (% edema and % larval survival)
which yielded an ECio of 11.36 mg Se/kg. The data for % lordosis indicate that it is a less
sensitive endpoint. The data for % hemorrhage showed no conclusive concentration-response
relationship: responses at concentrations 7.6 and 50.5 mg Se/kg dw varied widely without any
relationship to selenium concentration, precluding its use for estimating an ECio. The ECio value
of 11.36 mg/kg Se dw (combined larval survival and edema in response to Se concentration in
the parental ovaries) is considered an environmentally conservative chronic value for this
bluegill study. (See Appendix C for more discussion of this study).
The SMCV for bluegill reproductive endpoints based on ECio values is 17.95 mg Se/kg
dw in egg/ovary, based on the ECio values of Doroshov et al. (1992), Coyle et al. (1993), and
Hermanutz et al. (1992, and 1996 as corrected by Tao et al. 1999).
3.1.1.3.2 Micropterus salmoides (largemouth bass)
A laboratory study was conducted by Carolina Power & Light (1997) in which adult
largemouth bass obtained from a commercial supplier were fed an artificial diet spiked with a
gradient of selenomethionine concentrations for several months. Approximately 100 eggs from
each spawn were monitored for mortality and deformities up to the larval swim-up stage. The
authors combined survival and deformities into a single metric (i.e., survival as normal
offspring). The average concentration of selenium in the ovaries ranged from 3.1 mg/kg dw in
the control to 77.6 mg/kg dw in the high dietary treatment (53.1 mg/kg dw). The percent survival
of larval largemouth bass as a function of the selenium concentration in the parental ovary using
TRAP produced an ECio of 20.35 mg/kg dw and an EC20 of 23.60 mg/kg dw (Appendix C).
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3.1.2 Summary of Acceptable Studies of Fish Reproductive Effects
Table 3.1 summarizes the effect concentrations obtained from all acceptable reproductive
studies with fish. Summaries of the remainder of the reproductive studies (beyond the 4 most
sensitive genera described above) can be found in Section 6.1.2 below.
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Table 3.1. Maternal Transfer Reproductive Toxicity Studies.
Species
Acipemer
trammontanus
white sturgeon
Pimephales promelas
fathead minnow
Esox lucius
northern pike
Oncorhynchus mykiss
rainbow trout
Oncorhynchus clarkii
lew i si
Westslope cutthroat
trout
Oncorhynchus clarkii
lew i si
Westslope cutthroat
trout
Oncorhynchus clarkii
bouvieri
Yellowstone cutthroat
trout
Salvelinus malma
Dolly Varden
Reference
Linville 2006
Schultz and
Hermanutz
1990
Muscatello et
al. 2006
Holm 2002;
Holm et al.
2003, 2005
Rudolph et al.
2008
Nautilus
Environmental
2011
Formation
Environmental
2012
Golder 2009
Exposure route
dietary (lab)
dietary and waterborne
(mesocosm:
Monticello)
dietary and waterborne
(field: Saskatoon,
Sask.)
dietary and
waterborne
(field: Luscar River,
Alberta)
dietary and waterborne
(field: Clode Pond,
BC)
dietary and waterborne
(field: Clode Pond &
Fording River, BC)
dietary and waterborne
(field: Crow Creek,
Deer Creek, Sage
Creek, ID; hatchery:
Henry's Pond, ID)
dietary and
waterborne
(field: Kemess Mine
NW British Columbia)
Toxicological
endpoint
EC 10 for combined
edema and
deformities
LOEC for larval
edema and lordosis
EC 24 larval
deformities
EC 10 for skeletal
deformities
EC 10 for alevin
mortality
EC 10 for survival
at swim-up
EC 10 for survival
hatch through 1 5
days post swim up.
EC 10 for total
deformities
Chronic value,
mg/kg dwa
16.27 E
<23.85Ob'c
34.00 E
21.1Eb
24.11 E
24.02 E
25.25 E
56.22 E
SMCV
mg/kg dw
16.27 E
<23.85 O
34.00 E
21. IE
24.45 E
56.22 E
GMCV
mg/kg dw
16.27 E
<23.85 O
34.00 E
22.71 E
56.22 E
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Species
Salmo trutta
brown trout
Cyprinodon
macularius
desert pupfish
Lepomis macrochirus
bluegill
Lepomis macrochirus
bluegill
Lepomis macrochirus
bluegill
Micropterus salmoides
largemouth bass
Reference
Formation
Environmental
2011; AECOM
2012
Besser et al.
2012
Doroshov et al.
1992
Coyle et al.
1993
Hermanutz et
al. 1992, 1996
Carolina Power
& Light 1997
Exposure route
dietary and waterborne
(field: Lower Sage
Creek & Crow Creek,
ID)
dietary and waterborne
(lab)
dietary
(lab)
dietary and waterborne
(lab)
dietary and waterborne
(mesocosm:
Monticello) ^L
dietary (lab)
Toxicological
endpoint
EC 10 for larval
survival
Estimated ECio for
offspring survival
EC 10 larval edema
ECio for larval
survival
ECio for larval
edema
ECio for larval
mortality &
deformity
Chronic value,
mg/kg dwa
18.09 E
27 E
20.75 E
24.55 E
11.36Ob
20.35 O
SMCV
mg/kg dw
18.09 E
27 E
17.95 E
20.35 O
GMCV
mg/kg dw
18.09 E
27 E
17.95 E
20.35 O
E-Concentrationreported in egg; O- concentration reported in ovary
a All chronic values reported in this table are based on the measured concentration of selenium in egg/ovary tissues.
Tissue value converted from ww to dw. See Appendix C for conversion factors.
0 See Appendix E for an additional study results for fathead minnow.
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In order of their sensitivity to selenium, Table 3.2 presents the Genus Mean Chronic
Values from acceptable fish reproductive-effect studies that have been measured in terms of egg-
ovary concentrations.
Table 3.2. Ranked Genus Mean Chronic Values for Fish Reproductive Effects Measured as
Egg or Ovary Concentrations.
Rank
9
8
7
6
5
4
O
2
1
GMCV*
(mg Se/kg dw EO)
56.22
<34
27
<23.85
22.71
20.35
18.09
17.95
16.27
Species
Dolly Varden,
Salvelinus malma
Northern pike,
Esox lucius
Desert pupfish,
Cyprinodon macularius
Fathead minnow,
Pimephales promelas
Cutthroat trout,
Oncorhynchus clarkii
Rainbow trout,
Oncorhynchus mykiss
Largemouth bass,
Micropterus salmoides
Brown trout,
Salmo trutta
Bluegill sunfish,
Lepomis macrochirus
White sturgeon,
Acipenser transmontanus
SMCV
(mg Se/kg dw EO)
56.22
<34
27
<23.85**
24.45
21.1
20.35
18.09
17.95
16.27
* This table excludes Gambusia, which has a reproductive chronic value expressed as adult
whole-body rather than egg-ovary, because it is a live bearer.
** The fathead minnow SMCV is a conservative estimate because it does not include the higher
ECios for survival and deformities from GEI (2008), 35 - 65 mg/kg dw expressed as maternal
whole body, as noted in Appendix E, Figures E-l and E-2.
3.1.3 Invertebrate Chronic Effects
Below is a brief synopsis of the experimental design of the available invertebrate chronic
toxicity tests, and the resulting chronic values.
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3.1.3.1.1 Brachionus calyciflorus (rotifer)
Dobbs et al. (1996) exposed Brachionus calyciflorus to selenate in natural creek water for
25 days in a three-trophic level food chain test system. This is one of two laboratory-based
experiments (also see Bennett et al. 1986) that involved exposing algae to selenium (in this case
as sodium selenate) in water, and subsequently feeding the algae to rotifers which were in turn
fed to fish (fathead minnows). In the Dobbs et al. (1996) study, the rotifers and fish were
exposed to the same concentrations of sodium selenate in the water as the algae, but consumed
selenium bioaccumulated in the next lower trophic level. Rotifers did not grow well at
concentrations exceeding 108.1 jig Se/L in water, and the population survived only 6 days at
selenium concentrations equal to or greater than 202.4 jig Se/L in the water (40 jig Se/g dw in
the algae). Regression analysis of untransformed growth data (dry weight), determined 4 day
post-test initiation, resulted in a calculated ECio of 37.84 jig Se/g dw tissue.
3.1.3.1.2 Lumbriculus variegatus (oligochaete, blackworm)
Although not intended to be a definitive toxicity study for blackworms, Besser et al.
(2006) evaluated the bioaccumulation and toxicity of selenized yeast to the oligochaete,
Lumbriculus variegatus, which was intended to be used for dietary exposure in subsequent
studies with the endangered desert pupfish, Cyprinidon macularius. Oligochaetes fed selenized-
yeast diets diluted with nutritional yeast (54 to 210 mg Se/kg) had stable or increasing biomass
and accumulated Se concentrations as high as 140 mg/kg dw. The oligochaetes fed the undiluted
selenized-yeast (826 ug/g Se dry wt.) showed reduced biomass. The effect level is considered
>140 mg Se/kg dw.
^^
3.1.3.1.3 Centroptilum triangulifer (mayfly)
Mayfly larvae (Centroptilum triangulifer} were exposed to dietary selenium contained in
natural periphyton biofilms to eclosion (emergence) (Conley et al. 2009; Conley et al. 2011;
Conley et al. 2013). In Conley et al. (2009), the periphyton fed to the mayfly larvae were
exposed to dissolved selenite (radiolabeled 75Se) in November 2008 (12.6 and 13.9 |ig/L) and in
January 2009 ( 2.4, 2.4, 4.9, 10.3, and 10.7 |ig/L). Periphyton bioconcentrated Se an average of
1113-fold over the different aqueous selenium concentrations (see Table E-13 in Appendix E).
Twenty 4 to 6-day old mayfly larvae were exposed for 4.5 to 6 weeks to each of the periphyton
diets until the larvae eclosed to subimagos (final pre-adult winged stage). The subimagos were
allowed to emerge to the adult imago stage which deposited their egg masses in Petri dishes.
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Selenium concentrations were measured in postpartum adults along with their dry weights and
clutch size. Selenium increased in concentration from periphyton to the adult mayflies (trophic
transfer factor) an average of 2.2-fold. The authors observed a reduction in fecundity with diets
containing more than 11 mg Se/kg dw, which is considered the dietary threshold for this study.
Using the trophic transfer factor of 2.2, the periphyton selenium concentration of 11 mg/kg dw
translates to an adult mayfly selenium concentration of 24.2 mg/kg dw.
Conley et al. (2011) exposed larval C. triangulifer larvae similar to Conley et al. (2009)
to two different rations of periphyton (Ix and 2x) containing low, medium and high selenium
levels to evaluate the effect of feeding ration on the bioaccumulation of selenium and life cycle
performance of the mayfly. Mayfly larvae were fed either a Ix or 2x ration of periphyton loaded
with the three different selenium levels until the larvae eclosed to subimagos after 25-29 days.
Subimagos were induced to emerge to adults in petri dishes and their clutch size measured
through digital imaging. Mayflies fed the Ix ration had 54% and 72% reductions in survival
relative to controls in the medium and high Se treatment levels, respectively, both significant
(p<0.05). The mayflies fed the Ix ration also had significant reductions in fecundity in the low
(44% reduction), medium (63% reduction) and high (77% reduction) Se treatment levels.
However, for the mayflies that were fed the 2x ration, there were no significant differences
between the controls and any of the three Se treatment levels for any of the endpoints measured
including survival and fecundity. The 2x ration mayflies had 60% more biomass than the Ix
ration mayflies. This growth difference explains why the Ix ration mayflies had higher
concentrations of Se in their tissues (see Table E-14 in Appendix E). The two different rations
resulted in vastly different effect levels for Se, <12.8 mg/kg dw in the Ix ration test and >37.3
mg/kg dw in the 2x ration. It is apparent from this study that if the mayflies do not obtain
sufficient nutrition, they are more sensitive to selenium. Although reduced feeding levels occur
in nature, it is a confounding variable in this study that cannot be used to set a chronic effect
level for selenium.
Conley et al. (2013) evaluated the accumulation of selenite and selenate into periphyton
with a subsequent feeding exposure to mayfly larvae. As in the previous studies, C. triangulifer
larvae were fed periphyton previously exposed to different concentrations of selenium. In this
study, periphyton plates were first exposed to low (10 |ig/L) and high (30 |ig/L) concentrations
of either selenite or selenate and then fed to mayfly larvae to eclosion and to subimagos. The
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mean concentrations of selenium in the periphyton fed to the mayflies were 2.2, 12.8 and 37
mg/kg Se dw in the control, low and high treatments, respectively. Mayfly tissue (subimago)
concentrations (extrapolated from Figure 4a in Conley et al. 2013) were approximately 4-7, 20-
35, and 45-75 mg/kg Se dw, in the control, low and high treatments, respectively. The authors
reported significant reductions in survival from the control in the high Se treatment (both pooled
data and individual selenite and selenate treatments), but no significant differences were
observed in the low Se treatments. Secondary production (mayfly biomass) was significantly
reduced relative to the control in the high Se treatment for both selenium species. For the low Se
exposure treatments, secondary production was not significantly different than the control for the
selenite treated periphyton exposure, but was for the selenate and pooled data suggesting an
effect level between 20 and 35 mg/kg Se dw. These results as well as those observed in 2x ration
exposures in Conley et al. (2011) where no effects were observed at 37.3 mg/kg Se dw generally
support the chronic value determined for Conley et al. (2009) of 24.2 mg/kg Se dw. This
information included tabulated data from these studies presented in Appendix E.
3.1.4 Summary of Relevant Invertebrate Tests
The available measured invertebrate whole-body effect concentrations are shown in
Table 3.3. Because the intent of this assessment is to derive a concentration expressed in terms
offish tissue, Table 3.3 also provides information on how concentrations in invertebrate tissue
are translated (in Section 3.2) across media to predicted WB fish tissue concentrations (Trophic
Level 3, TL3) in a system having invertebrates and fish. That is, consistent with the
bioaccumulation modeling approach of Section 3.2, the second column of Table 3.3 uses the
median trophic transfer factor of 1.27 from Table 3.11 to yield expected WB fish tissue
concentrations in a system having invertebrates and fish. Whether comparing TL2 (invertebrate)
whole-body GMCVs directly to Table 3.5 TL3 (fish) whole-body GMCVs, or via the trophic
transfer adjustment in the second column of Table 3.3, it is apparent that invertebrates are not
among the more sensitive species.
The relative insensitivity of invertebrates when compared with the fish whole-body
concentrations demonstrates that invertebrates are generally protected by selenium criterion
values derived from fish. Therefore, the invertebrates are considered implicitly in the species
sensitivity distribution, and are counted toward the number of values available to calculate the
fish tissue criterion elements (as egg-ovary, whole-body, and muscle), and the missing
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invertebrate MDRs (4 and 5) are considered satisfied by the available invertebrate data.
Table 3.3. Ranked Invertebrate Whole-Body Chronic Values with Translation to Expected
Accompanying Fish Whole-Body Concentrations
SMCV & GMCV
as measured
(Trophic Level 2)
(mg Se/kg dw WB)
>140
37.84
24.2
Accompanying Trophic Level 3
Median Whole-Body Concentration
Predicted by Bioaccumulation
Model (Section 3.2)
(mg Se/kg dw WB TL3)
> 178
48.1
30.7
Species
Oligochaete, black,
Lumbriculus variegatus
Rotifer,
Brachionus calyciflorus
Mayfly,
Centroptilum triangulifer
3.1.5 Derivation of Tissue Criterion Element Concentrations
Data used to derive the final chronic value were differentiated based on the effect
(reproductive and non-reproductive effects). Acceptable chronic toxicity data on fish
reproductive effects are available for 10 fish genera. Acceptable chronic toxicity data on non-
reproductive effects are available for 7 fish genera and 3 invertebrate genera. The fish non-
reproductive effects data were not used to calculate tissue criterion elements because they were
more variable and less reproducible than the data on reproductive effects. The SSD is
predominantly populated with data on fish species because field evidence demonstrated that fish
communities were affected in situations having no observable change in the accompanying
diverse array of invertebrate communities. As a result, decades of aquatic toxicity research have
focused primarily on fish. The studies that have been done with invertebrates (Table 3.3, Section
3.1.3) have shown them to be more tolerant than most of the tested fish species. While
potentially sensitive due to physiologic similarities to fish, amphibian effects clearly attributable
to selenium are largely unknown (Unrine et al. 2007; Hopkins et al. 2000; Janz et al. 2010).
Hopkins et al. (2000) reported that amphibian larvae at sites receiving coal combustion wastes
appear to efficiently accumulate selenium in their tissues and possibly due to selenium have
exhibited axial malformations. In a recent laboratory exposure, Masse et al. (2014) determined
an ECio of 24.8 mg/kg Se for the African clawed frog (Xenopus laevis) suggesting that
anphibians have a similar sensitivity to fish (see Section 6.1.4).
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3.1.5.1 Fish Egg-Ovary Concentration
The lowest four GMCVs from Table 3.2 are shown below in Table 3.4.
Table 3.4. Four lowest Genus Mean Chronic Values for Fish Reproductive Effects.
Relative Sensitivity
Rank
4
3
2
1
Genus
Micropterus
Salmo
Lepomis
Acipemer
GMCV
(mg Se/kg dw egg-ovary)
20.35
18.09
17.95
16.27
With N=15 GMCVs (see Section 3.1.6), the 5* percentile projection yields an egg/ovary
criterion of 15.8 mg Se/kg dw egg/ovary, lower than the most sensitive fish species tested, white
sturgeon (A. transmontanus). The egg/ovary criterion element concentration is compared to the
distribution of egg/ovary chronic values in Figure 3.1.
fi4
1
5
*?. 39 -
|
> 16
0
*
.c
U)
LL.
C
A
X
X
) 2 4 6 8 1
Rank
A Salvelinus
X Esox
X Cyprinodon
• Pimephales
+ Oncorhynchus
Micropterus
Salmo
+ Lepomis
• Acipenser
... EOFCV
0
Figure 3.1. Distribution of reproductive-effect GMCVs for fish measured as egg-ovary
concentrations.
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3.1.5.2 Fish Whole-Body Criterion Element Concentration
Using the egg-ovary to whole-body conversion factors of the bioaccumulation modeling
approach presented subsequently in Section 3.2, Table 3.5 shows the conversion of
reproductive-effect egg-ovary concentrations to whole-body concentrations. It can be seen that
for some species, the conversion was done in a single step, using a measured egg-ovary (EO) to
whole-body (WB) ratio specific to the taxon. But for other species, it was done in two steps, first
converting to muscle (M) and then applying a generic M/WB ratio. There were sufficient egg-
ovary and whole-body selenium measurements to directly derive CF values for some species,
and matched pairs of selenium measurements in muscle and whole body were also available.
When paired tissue data used to derive CF values were not available, the EPA estimated missing
data using a hierarchical approach based on taxonomic relatedness to estimate CF. This approach
is consistent to that used to derive TTF estimates, and is described in Section 3.2.2, and in much
greater detail in Appendix B (see Table B-6 for final CF values).
Table 3.5. Tested Reproductive-Effect Egg-Ovary (EO) Concentrations Converted to
Whole-Body (WB) Concentrations.
Taxon*
Salvelinus
Esox
Cyprinodon
Pimephales
O. mykiss
O. clarkii
Oncorhynchus
Micropterus
Salmo
Lepomis
EO
Chronic
Value
56.22
34.00
27.00
23.85
21.10
24.45
22.71
20.35
18.09
17.95
EO/WB
Ratio
1.611
2.389
1.210
1.997
2.441
1.964
2.190
1.419
1.446
2.133
Calculated
WB Repro
Chronic
Value
34.90
14.23
22.31
11.94
8.64
12.45
10.37
14.34
12.51
8.41
Basis for EO/WB Ratio
(from Appendix B)
Median Dolly Varden EO/M (1.264) x
median fish M/WB (1.274)
Median northern pike EO/M (1.875) x
median fish M/WB (1.274)
Median desert pupfish EO/WB
Median Cyprinidae EO/WB
Median rainbow trout EO/M (1.916) x
median fish M/WB (1.274)
Median cutthroat trout EO/M (1 .805) x
median fish M/WB (1.274)
Using geomean of species ratios yields
geomean of SMC Vs
Median Centrarchidae EO/WB
Median brown trout EO/WB
Median bluegill EO/WB
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Taxon*
Acipenser
EO
Chronic
Value
16.27
EO/WB
Ratio
1.694
Calculated
WB Repro
Chronic
Value
9.60
Basis for EO/WB Ratio
(from Appendix B)
Median white sturgeon EO/M (1.330) x
median fish M/WB (1.274)
* The GMCV for Gambusia, a live bearer not shown in the above conversion table, was
originally measured as adult WB, not EO, and is >13.38 mg Se/kg dw WB. The "greater than"
sign signifies that no effects were found at the studies' highest observed concentrations.
Table 3.6. The lowest four reproductive-effect whole-body GMCVs.
Relative
Sensitivity
Rank
4
O
2
1
Genus
Pimephales
Oncorhynchus
Acipenser
Lepomis
GMCV
(mg Se/kg dw whole-body)
11.94
10.37
9.60
8.41
Because the factors used to convert egg-ovary to whole-body concentrations vary across
species, the whole-body rankings differ from the egg-ovary rankings. With N=15 GMCVs, the
5th percentile projection yields a whole body criterion of 8.0 mg Se/kg dw whole-body, slightly
lower than the most sensitive fish species tested, bluegill (Lepomis macrochirus). The fish whole
body criterion is compared to the distribution offish whole-body reproductive chronic values in
Figure 3.2
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fid 00 -i
•D
MOT no -
01
1/1
00
u
3
CO
S o nn
.c
tu>
A nn
c
X
+ • *
i • * ~
1 i i i k
) 2 4 6 8 10
Rank
A Salvelinus
X Cyprinodon
• Micropterus
+ Esox
Salmo
— Pimephales
* Oncorhynchus
• Acipenser
A Lepomis
_ _ _ VVB FCV
Figure 3.2. Distribution of reproductive-effect GMCVs for fish, measured as egg-ovary
concentrations but converted to whole-body concentrations as shown in Table 6.
(No conversion was done for live-bearer Gambusia, originally measured as WB).
3.1.5.3 Fish Muscle Criterion Element Concentration
Using the egg-ovary to muscle conversion factors of the bioaccumulation modeling
approach (presented later in Section 3.2), Table 3.7 shows the conversion of reproductive-effect
egg-ovary concentrations to whole-body concentrations. For all but Cyprinodon (desert pupfish),
the conversion was completed in a single step, applying an EO/M ratio specific to the taxon.
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Table 3.7. Tested Reproductive-Effect Egg-Ovary (EO) Concentrations Converted to
Muscle (M) Concentrations.
Taxon
Salvelinus
Esox
Cyprinodon
Pimephales
O. mykiss
O. clarkii
Oncorhynchus
Micropterus
Salmo
Lepomis
Acipenser
EO
Chronic
Value
56.22
34
27
23.85
21.1
24.45
22.71
20.35
18.09
17.95
16.27
EO/M
Ratio
1.264
1.875
0.950
1.590
1.916
1.805
1.860
1.187
1.135
1.375
1.330
Calculated
Muscle Repro
Chronic
Value
44.46
18.13
28.43
15.00
11.01
13.55
12.21
17.14
15.94
13.05
12.23
Basis for EO/M Ratio (from
Appendix B)
Median Dolly Varden EO/M
Median northern pike EO/M
Median desert pupfish EO/WB (1.210)
/ Median fish M/WB (1 .274)
Median Cyprinidae EO/M
Median rainbow trout EO/M
Median cutthroat trout EO/M
Using geomean of species ratios yields
geomean of SMC Vs
Median Genus Micropterus EO/M
Median brown trout EO/WB
/ median fish M/WB
Median bluegill EO/M
Median white sturgeon EO/M
Table 3.8. The lowest four reproductive-effect fish muscle GMCVs.
Relative Sensitivity
Rank
4
3
2
1
Genus
Pimephales
Lepomis
Acipenser
Oncorhynchus
GMCV
(mg Se/kg dw muscle)
15.00
13.05
12.23
12.21
Because the factors used to convert egg-ovary to muscle concentrations vary across
species, the whole-body rankings differ from both from the egg-ovary rankings and the muscle
rankings. With N=15 GMCVs, the 5th percentile projection yields a muscle criterion of 11.3 mg
Se/kg dw muscle, lower than the muscle value for the most sensitive fish genus tested,
Oncorhynchus. Figure 3.3 compares the fish muscle criterion to the distribution offish muscle
reproductive chronic values in Table 3.7.
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CA nn -
•D
•* 39 nn
01
1/1
no
J ic nn
3
01
(J
U)
3 s nn
.c
w
LL.
A nn
C
A
X
+ *
4-I-+— 1
) 2 4 6 8 10
Rank
A Salvelinus
X Cyprinodon
X Gambusia
• Esox
+ Micropterus
- Salmo
— Pimephales
* Lepomis
• Acipenser
_ _ _ M FCV
Figure 3.3. Distribution of reproductive-effect GMCVs for fish, measured as egg-ovary
concentrations but converted to muscle concentrations as shown in Table 3.7.
(Live-bearer Gambusia was converted from WB to muscle).
3.1.6 Selenium Fish Tissue Toxicity Data Fulfilling Minimum Data Needs
The toxicity data currently available for genera and species fulfilling the U.S.EPA
Ambient Water Quality Criteria Guidelines recommendations for calculation of the freshwater
chronic criterion are described in Section 3.1.1, 3.1.3, 6.1.2 and Appendix C and summarized in
Table 3.9
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Table 3.9. Minimum Data Requirements Summary Table Reflecting the Number of Species
and Genus Level Mean Values Represented in the Chronic Toxicity Dataset for Selenium
in Freshwater.
Freshwater Minimum Data Requirement
1 . Family Salmonidae in the class Osteichthyes
2. Second family in the class Osteichthyes,
preferably a commercially or recreationally
important warmwater species
3. Third family in the phylum Chordata (may be
in the class Osteichthyes or may be an
amphibian, etc.)
4. Planktonic Crustacean
5. Benthic Crustacean
6. Insect
7. Family in a phylum other than Arthropoda or
Chordata (e.g., Rotifera, Annelida, or Mollusca)
8. Family in any order of insect or any phylum
not already represented
Total
Genus Mean Chronic
Value (GMCV)
3
2
5>
See text
See text
1
1
1
15
Species Mean Chronic
Value (SMCV)
4
2
5
See text
See text
1
1
1
16
The first three of these MDRs in Table 3.9 are easily fulfilled by the fish species
represented in Sections 3.1.1, 6.1.2 and Appendix C. Because the field observations of
contaminated sites have found effects on fish and birds in the absence of changes in invertebrate
assemblages, scientific studies on the chronic toxicity of dietary selenium for invertebrates has
been very limited. The few dietary chronic toxicity studies that are available for invertebrate
species (arthropods , rotifers, and worms) indicate that they less sensitive than fish, with
available data indicating invertebrate whole body mean chronic values ranging from
approximately 3 to 12 times higher than the fish whole body criterion value recommended in this
document (data provided in Section 3.1.3). The above invertebrate data address MDRs 6-8,
leaving only MDRs 4 and 5, for the planktonic and benthic crustaceans, to be addressed. Because
the 5* percentile calculation methods for the FCV use actual numeric values for the GMCVs of
only the four most sensitive (fish) genera in the selenium dataset, it is only necessary to know
that the more tolerant genera have GMCVs that are greater than those of the low four. A
recommendation in the draft white paper on Aquatic Life Criteria for Contaminants of Emerging
Concern Part I (U.S. EPA 2008b), which was supported by the Science Advisory Board, states
"because only the four most sensitive genus mean chronic values (GMCVs) are used in the
criterion calculations, chronic testing requirements for a taxon needed to meet an MDR should
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be waived if there is sufficient information to conclude that this taxon is more tolerant than the
four most sensitive genera."
Currently, there are no available data on the chronic toxicity to crustaceans via dietary
exposure to selenium. Since there are data available for insects (Centroptilum spp. mayfly), EPA
has used the taxonomic association at the level of phylum (Arthropoda) to allow insects to act as
a surrogate for crustaceans. There is also associative evidence that macroinvertebrates in general
are less sensitive than fish. At sites where there have been documented effects to fish and
aquatic-dependent birds from selenium exposure (e.g., Kesterson Reservoir, Belews Lake, Hyco
Reservoir), field observations and data indicate that there has been no evidence of effects to
macroinvertebrates including crustaceans (Janz et al. 2010). In addition, Janz et al. (2010) notes
that the key vector for selenium toxicity via maternal transfer is selenium loading in the egg via
vitellogenesis. Crustaceans, and other arthropods are not known to deposit a significant amount
of vitellogenin in the egg compared with oviparous vertebrates like fish, therefore, less selenium
is likely transferred to the egg via deposition of vitellogenin. These mechanistic considerations
are thus consistent with the absence of observed field effects on aquatic macroinvertebrates,
including crustaceans and other arthropods, and with the Chapman et al. (2009, 2010) expert
consensus that it is the egg-laying vertebrates that are most at risk.
Applying this concept to the selenium criterion 5* percentile calculations, GMCVs for
MDRs 4 and 5 (the two crustacean MDRs) should be waived and counted in the total number of
GMCVs in the dataset, based on (a) the difference in the measured effect values discussed above,
and (b) the lack of observed invertebrate field effects linked to selenium (for example, as
concluded by Lemly 2002, pages 21-23, and Janz et al. 2010). Thus data are adequate to fulfill
the data needs for developing a chronic selenium criterion.
The total number of GMCVs available to derive the chronic criterion is 154. These
include ten fish genera from Sections 3.1.1 and 6.1.2 (Acipemer, Salmo, Lepomis, Micropterus,
Oncorhynchus, Pimephales, Gambusia, Esox, Cyprinodon, and Salvelinus) [Added to these are
the tested invertebrate genera Centroptilum, Brachionus, and Lumbriculus from Section 3.1.3],
and lastly the two waived genera for MDRs 4 and 5 (crustaceans).
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3.2 CHRONIC WATER COLUMN-BASED SELENIUM CRITERION ELEMENT
3.2.1 Translation from Fish Tissue Concentration to Water Column Concentration
EPA derived the chronic water column selenium criterion element by translating the egg-
ovary concentration to an equivalent water concentration. EPA worked with the United States
Geological Survey to derive a translation equation that utilizes a mechanistic model of
bioaccumulation previously published in peer-reviewed scientific literature (Luoma et. al., 1992;
Wang et. al., 1996; Luoma and Fisher, 1997; Wang, 2001; Schlekat et al. 2002b; Luoma and
Rainbow 2005; Presser and Luoma 2006; Presser and Luoma 2010; Presser 2013). This model
quantifies bioaccumulation in animal tissues by assuming that net bioaccumulation is a balance
between assimilation efficiency from diet, ingestion rate, rate of direct uptake in dissolved forms,
loss rate, and growth rate. The basic model is given as:
Where:
water
ku
AE
IR
Wo + g> (Equation 1)
Concentration of metal in water (|ig/L)
Average concentration of metal in all tissues at steady-state (|ig/g)
Efflux rate (/d)
Growth rate (/d)
Uptake rate (L/g-d)
Assimilation efficiency (%)
Ingestion rate (g/g-d)
Concentration in food (|ig/g)
3.2.1.1 Simplifying the Bioaccumulation Model
Specific application to selenium bioaccumulation permits the simplification of Equation 1
in two ways. One simplification is removing the parameter representing growth rate (g), and the
other simplification is removing the parameter representing direct aqueous uptake (&„).
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3.2.1.1.1 Growth Rate
The growth rate constant g is included in Equation 1 because the addition of body tissue
has the potential to dilute the concentration of bioaccumulative chemicals when expressed as
chemical mass per tissue mass. For very hydrophobic chemicals with low excretion rates such as
polychlorinated biphenyls, growth can be an important factor in bioaccumulation estimates
(Connolly and Pedersen 1988). However, Luoma and Rainbow (2005) suggest that for selenium,
growth rate is a relatively inconsequential parameter under most circumstances. Food
consumption is typically high during periods of high growth rate. Because food consumption is
the primary route of selenium uptake in aquatic organisms (Ohlendorf et al. 1986a, b; Saiki and
Lowe 1987; Presser and Ohlendorf 1987; Lemly 1985a; Luoma et al. 1992; Presser et al. 1994,
Chapman et al. 2010), high consumption rates of selenium-contaminated food may counteract
the selenium dilution that occurs with the addition of body tissue during periods of fast growth.
EPA evaluated the effect of removing the parameter g in the Equation 1 by performing a
sensitivity analysis. EPA analyzed a series of hypothetical tissue concentration estimates using
Equation 1 with g ranging between 0 (no growth) and 0.2/day (a relatively high rate of growth).
In one analysis, tissue concentrations of selenium were estimated using static values of IR. In a
second analysis, tissue concentrations of selenium were estimated using values of IR that were
adjusted for growth rate using a method similar to the approach used in a model of organic
chemical accumulation in aquatic food webs (Thomann et al. 1992). As expected, estimates of
selenium tissue concentrations were significantly reduced at progressively higher growth rates
when IR remained constant. However, selenium concentrations remained fairly steady or slightly
increased with progressively higher growth rates when IR was adjusted for the bioenergetics of
growth. This analysis supports the hypothesis that a higher//? (and consequently greater rate of
selenium ingestion) associated with the higher bioenergetic requirements of rapidly growing
young fish tends to oppose the dilution of selenium in their tissues due to growth, whereas a
lower//? (and consequently lower rate of selenium ingestion) associated with the lower
bioenergetic requirements of slower growing older fish tends to oppose the bioconcentration of
selenium in their tissues. EPA concludes from this analysis that omitting the growth rate
parameter g is an appropriate simplification of Equation 1. A more detailed description of this
sensitivity analysis is provided in Appendix J.
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3.2.1.1.2 Uptake Rate
The uptake rate constant ku is included in Equation 1 to account for direct absorption of
bioaccumulative chemicals in the dissolved phase. However, dietary intake of selenium is the
dominant source of exposure, suggesting that ku may also be relatively inconsequential for
selenium accumulation (Luoma and Rainbow 2005). Because aqueous uptake of selenium makes
up a small percentage of bioaccumulated selenium (Fowler and Benayoun 1976; Luoma et. al.,
1992; Roditi and Fisher 1999; Wang and Fisher 1999; Wang 2002; Schlekat et. al., 2004; Lee et.
al., 2006), Presser and Luoma (2010a, 2010b, 2013) deemed removal of ku from Equation 1 as an
acceptable simplification.
EPA evaluated the effect of removing the parameter ku in the Equation 1 by performing a
sensitivity analysis. EPA analyzed a series of tissue concentration estimates using Equation 1 and
a realistic range of ku values for trophic level 2 and trophic level 3 organisms. The analysis
suggests that approximately 75% of selenium exposure in trophic level 2 organisms
(invertebrates) and over 90% of selenium exposure in trophic level 3 organisms occurs through
consumption of selenium-contaminated food. EPA concluded that omitting the aqueous uptake
rate constant ku is an appropriate simplification of Equation 1. A more detailed description of this
sensitivity analysis is provided in Appendix J.
3.2.1.1.3 Derivation of the Translation Equation
Disregarding growth (g) and uptake of selenium dissolved in water (ku x Cwater), Equation
1 becomes Equation 2 (Reinfelder et al. 1998):
AExIRxCfood
or:
AExIR
X ^
tissue , food
e (Equation 2)
Because application of the bioaccumulation model applies to a single species, the
AExIR
combination of species-specific physiological parameters expressed as remains
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AExIR
constant for the species. Thus the EPA defines the expression as a single species-
ke
specific Trophic Transfer Function (TTF) given as Equation 3 (Reinfelder et al. 1998):
AExIR
TTF =•
e (Equation 3)
AExIR
Substituting TTF for in Equation 2 yields:
k
Ctissue = TTF x C food (Equation 4)
The trophic level of the organisms considered can be denoted by superscripts given as:
^iTL2 _
ttssue " A ^f°od (Equation 5)
Ctjssue as defined here represents the steady-state proportional concentration of selenium in the
tissue of trophic level 2 organisms relative to the concentration of selenium in their food source.
Using the same rationale, the average concentration of selenium in the tissues of trophic
level 3 organisms can be expressed as the concentration of selenium in its food multiplied by a
TTF which is given as:
^TL3 _ jvrr>TL3 r
-------
Defining the term c^ue as the concentration of selenium in fish tissue, defining the term
Cfood as the concentration of selenium in living and nonliving particulate material ingested by
invertebrates, and expressing the product of all TTF values as a single term results in the
equation:
n - j^rv composite ^ fFmiatirm Q"l
^whole-body iil ^^particulate ^E,qUctLlUll 7J
where:
C particulate = the concentration of selenium in particulate material
^hoie-body = the concentration of selenium in the whole body offish
TTFcomposie = the product of all trophic transfer function values
Equation 9 quantitatively expresses selenium bioaccumulation in fish (Cwhoie-body) as the
product of the concentration of selenium at the base of the food web (Cparticuiate) and a parameter
representing the trophic transfer of selenium through all dietary pathways (7777*°™?°^) This
model of bioaccumulation is conceptually similar to the model of bioaccumulation utilizing a
bioaccumulation factor (BAF). A BAF is the ratio of the concentration of a chemical in the tissue
of an aquatic organism to the concentration of the chemical dissolved in ambient water at the site
of sampling (USEPA 200 Ic). Similar to the term fTF00^0^, a BAF quantitatively represents the
relationship between the chemical concentrations in multiple environmental compartments.
However, a BAF is empirically derived from site-specific measurements, whereas jj'pcomPosl-te js
derived from knowledge of the ecological system. Because each TTF is associated with a
particular taxon, 777^°™?°^ can be inferred for an aquatic system using existing knowledge and
reasonable assumptions, without the considerable time and cost of collecting and analyzing
tissue and water samples.
Equation 9 characterizes the bioaccumulation of selenium as a combination of TTF
parameters from all steps in the dietary pathway of the predator species of interest. Thus it is
possible to differentiate bioaccumulative potential for different predator species and food webs
by modeling different exposure scenarios. For example, where the fish species of interest is a
trophic level 4 predator that primarily consumes trophic level 3 fish, the term 777^°™?°^ can be
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represented as the product of all TTF parameters that includes the additional trophic level given
as:
10)
where:
TT 9
TTF = the trophic transfer function of trophic level 2 species
y transfer function of the trophic level 3 species
= the trophic transfer function of the trophic level 4 species
TTF°mposlte = the product of all trophic transfer functions
^^^
Similarly, the consumption of more than one species of organism at the same trophic
level can also be modeled by expressing the TTF at a particular trophic level as the weighted
average of the TTFs of all species consumed given as:
TTF TLx = Y, (TTF^ x w;. ) (Equation 1 1)
where:
7
TTFi = the trophic transfer function of the i* species at a particular trophic level
w; = the proportion of the ith species consumed
These concepts can be used to formulate an expression of TTF°omposlte to model selenium
bioaccumulation in ecosystems with different consumer species and food webs. Figure 3.4
describes four example food web scenarios and the formulation of j"TFcomP°slte to model selenium
bioaccumulation in each of them.
The parameter jj'pcomPosl-te quantitatively represents all dietary pathways of selenium
exposure for a particular fish species within an aquatic system. The parameter is derived from
species-specific TTF values representing the food web characteristics of the aquatic system, w,,
the proportion of species consumed. See text for further explanation.
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A) Three trophic levels (simple):
rjirrrrj
''•?
'
B) Four trophic levels (simple);
mposite
J>pTL3
C) Three trophic levels (mix within trophic levels);
TTF00"""'"" = TTFT" x
Wl)+ (jTF2TL2
W] J;
w2 »'
D) Three trophic levels (mix across trophic levels);
W,
w
E) Four trophic levels (mix across trophic levels);
1 t
Figure 3.4. Example aquatic system scenarios and the derivation of the equation parameter
TTFcomposite.
Because EPA's objective is to derive an equation that translates a fish tissue
concentration of selenium to a water column concentration, the term Cwater is reintroduced into
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Equation 9 by defining the enrichment function EF representing the steady state proportional
bioconcentration of dissolved selenium at the base of the aquatic food web given as:
c
T-T T-T particulate
water (Equation 12)
where:
C particulate = Selenium concentration in particulate material (|ig/g)
CWater = Concentration of selenium dissolved in water (|ig/L)
EF = Enrichment function (L/g)
Rearranging the terms of Equation 12:
C particulate = EF X Cwater (Equation 13)
Substituting EFxCwater for c articu!ate m Equation 9 results in:
C» 'I vi >l 'composite T^T^ ^ I--T* .• * *\
whole-body =TTF xEFxCwater (Equation 14)
Solving for the concentration of selenium in water in Equation 14 results in:
^whole-bod
water 717177 composite .,7777 ,„ .,.,-N
*•*•? xM (Equation 15)
Because Equation 15 relates a concentration of selenium in water to the concentration of
selenium throughout all tissues of the body, Cwh0ie-body must be converted to an equivalent
concentration in eggs or ovaries. The EPA achieved this conversion by incorporating a species-
specific conversion factor (CF) into Equation 15. CF represents the species-specific proportion
of selenium in egg or ovary tissue relative to the concentration of selenium in all body tissues
and is given as:
CF=C-
r
whole-body (Equation 16)
Where:
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CF = Whole-body to egg-ovary conversion factor (dimensionless ratio).
= Selenium concentration in the eggs or ovaries offish (|ig/g)
= Selenium concentration in the whole body offish (|ig/g).
Rearranging the terms of Equation 16 yields:
C
_ egg-ovary
whole-body
(Equation 17)
Substituting Cwh0ie_body in Equation 15 with —egg °™y yields the translation equation:
CF
^^^
n _ egg-ovary
WU,LVt rr-irr-iT—i COWLVOSllQ T^ T^ /""< T"1
TTF p xEFxCF (Equation 18)
where TTFcomposlte equals the product of all trophic transfer functions from trophic level 2
through the target fish species.
Equation 18 describes an ecosystem-dependent relationship between the concentration of
selenium in the eggs and ovaries offish with the concentration of selenium in the water column.
This approach explicitly recognizes the sequential transfer of selenium between environmental
compartments (water, paniculate material, invertebrate tissue, fish tissue, and eggs and/or ovary
tissue) by incorporating quantitative expressions of selenium transfer from one compartment to
the other. Because this approach uses food web modeling along with species-specific TTF and
CF parameters to quantify most of the transfer between compartments, however, the only field
measurements needed to relate selenium in egg-ovary and water are measurements from the
water column and particulate material sufficient to calculate EF.
3.2.2 Equation Parameters
Empirical or laboratory data related to selenium bioaccumulation in aquatic organisms
are needed to derive the equation parameters EF, TTF, and CF. EPA obtained data from
published literature as described above The search resulted in the retrieval of 54 acceptable
studies containing a total of 7,203 selenium measurements at 610 aquatic sites (2,584 from
water, 289 from algae, 30 from detritus, 808 from sediment, 1,137 from various species of
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invertebrates, and 2,355 from various species offish) and 34 acceptable studies yielding 139
physiological constants (48 values of ke, 81 values of AE, and 10 values ofIR). EPA used this
collection of selenium measurements to calculate site-specific EF values and develop species-
specific TTF and CF values in an unbiased and systematic manner. A more detailed description
of how EPA calculated EF is described below. A more detailed description of how EPA
calculated TTF and CF is described in Appendix B.
3.2.2.1 Derivation of Trophic Transfer Function (TTF) Values
EPA derived T7F values for taxonomic groups of invertebrates and fish by either using
physiological coefficients found in the literature, or by evaluating the empirical relationship
between matched pairs of selenium measurements in organisms and the food they consumed.
When physiological coefficients were available, EPA calculated a T7F value using the equation:
k
e
Where:
k
e = Elimination rate constant (/d)
AE = Assimilation efficiency (%)
IR = Ingestion rate (g/g-d)
EPA also derived TTF values using empirical measurements of selenium from field
studies. EPA searched its collection of available selenium measurements and identified
measurements taken from aquatic organisms. For each measurement from an aquatic organism,
EPA searched for additional measurements from other aquatic organisms or particulate material
that was collected from the same aquatic site and of a type deemed likely to be ingested as a food
source or in conjunction with feeding activity (i.e., a lower trophic level). If multiple lower
trophic level measurements were matched to an aquatic organism measurement, the median of
the lower trophic level measurements was calculated. Each pair of measurements, one taken
from an aquatic organism and the other taken from lower trophic level organisms or particulate
material, was designated as a matched pair.
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Because selenium is transferred to aquatic animals primarily through aquatic food webs,
the observable concentration of selenium in different environmental compartments may vary
over time. To establish an appropriate time period with which to define matched pairs of
selenium measurements, the effect of sample collection time on the relationship between
selenium concentrations in different media was analyzed. EPA defined matched pairs of
selenium measurements as described above using different relative collection time ranges and
estimated the strength of the relationship between the two measurements by calculating the
Pearson product-moment correlation coefficient (r).
Figure 3.5 shows the correlation coefficients for selenium measurements taken from the
same aquatic sites when the measurement collection times were systematically shifted relative to
one another. Each correlation coefficient was calculated from a set of data within a specified
range of relative collection times with respect to the higher trophic level. For example, the
correlation coefficient calculated from paniculate and invertebrate measurements with a relative
sample collection time of 30 to 60 days were from invertebrate and paniculate samples collected
at the same site, with the invertebrate samples collected 30 to 60 days after the particulate
samples. Similarly, the correlation coefficient calculated from paniculate and invertebrate
measurements with a relative collection time of-60 to -30 days were from invertebrate and
particulate samples that were collected at the same site, with the invertebrate samples collected
30 to 60 days before the particulate samples.
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Particulate versus invertebrate
'1
r o
-1 -
IMi V'i*****"******,
<
rji
I
<
rJi?l
<
1
(
i
•
i n
1
<
i
Invertebrate versus fish
'•
f....i* i *.* _*_„,
i i
•
i -^
;* i •
-1
ooooooooo222°~rtrtrt00000000000000000000
000000000*??oOOs2)^^^°°°°°°°°°°°°°°°°°°
OJO
-------
and/or bioaccumulation characteristics that require different relative collection time criteria to
accurately characterize selenium relationships.
After matched pairs of selenium measurements from samples collected in the field were
identified, EPA evaluated two different analytical approaches to derive species-specific TTF
values from them. TTF was previously defined above as the steady state proportion relating the
concentration of selenium in the tissue of aquatic organisms to the concentration of selenium in
the food they ingest such that:
Ctissue =TTFxC food (Equation 4)
Rearranging the terms of Equation 4 yields:
C
rrrT~C< tissue
~ r
^ food
Because TTF can be defined as the ratio of the concentration of selenium observed in the
tissue of an aquatic organism to the concentration of selenium observed in the tissue or material
the organism ingests, one approach for deriving TTF values from field data is to simply use the
ratio of the two values. EPA evaluated this approach by calculating the ratios for all matched
pairs of selenium measurements, and for each species or taxonomic group, used a statistic of
central tendency of the distribution of ratios as the TTF value. An advantage of quantifying the
relationship between selenium in two environmental compartments using ratios is that it is a
simple and straightforward method that is conceptually similar to a bioaccumulation factor
(BAF). A disadvantage of this approach is that it presumes that the quality and quantity of data
used to derive the ratios adequately represent the relationship being characterized. Furthermore,
many aquatic organisms tend to bioaccumulate more metals at low environmental concentrations
(McGeer et al. 2003; Borgman et al. 2004; DeForest et al. 2007; USEPA 2007). Thus a
distribution of ratios could be biased toward larger values if the data are obtained from aquatic
systems with low selenium concentrations.
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Another analytical approach for deriving TTF values from matched pairs of selenium
measurements is to model the species-specific relationships using linear regression. One
possibility is to regress the concentration of selenium in the food of a particular species or
taxonomic group with the concentration of selenium in the organism's tissue, and use the
regression coefficient as the TTF. EPA evaluated this approach by applying ordinary least
squares (OLS) linear regression on the matched pairs of data. The regression coefficient (slope of
the fitted line) was then taken as the TTF value for that species or taxonomic group. An
advantage of this regression approach is that it estimates the quantitative relationship of selenium
across a range of environmental concentrations in a manner that allows statistical assessment.
Disadvantages of this regression approach include the assumption that the underlying data are
normally distributed; one or a few very high values can have a disproportionate influence on the
slope of the fitted line; and the bioaccumulation model does not account for a non-zero y-
intercept. Constraining the y-intercept to zero (also known as regression through the origin or
RTO) eliminates the added complexity of a non-zero y-intercept. However, RTO further
increases the disproportionate influence of one or a few high values on the slope of the fitted
line. Furthermore, RTO does not provide a straightforward way of evaluating goodness of fit
(Gordon 1981).
After evaluating both approaches, EPA decided to use a hybrid approach by designating
the median of the ratio of matched pairs of selenium measurements as the TTF value, but only if
OLS linear regression of those data resulted in a significant (P < 0.05) fit and positive regression
coefficient. Requiring a significant positive OLS linear regression coefficient confirms the
relationship between selenium in organisms and the food they ingest is adequately represented
by the available data. Using the median of the individual ratios provides an estimate of central
tendency for that relationship that is less sensitive to potential bias from measurements taken
from aquatic systems with very high or very low selenium concentrations. Some aquatic
organisms exhibit selenium bioaccumulation inversely related to water concentration (McGeer et
al. 2003; Borgman et al. 2004; DeForest et al. 2007). This inverse relationship is likely due to
saturation uptake kinetics of specific transport mechanisms that regulate metals bioaccumulation
within certain ranges (USEPA 2007). EPA evaluated the effect of very high and very low
selenium concentrations on the calculation of TTF values using the hybrid approach described
above by excluding selenium measurements above various minimum and/or below various
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maximum selenium concentrations. EPA found that using the median ratio effectively attenuates
any effects of selenium concentration on the calculation of TTF values using the hybrid approach
described above without the need to introduce additional arbitrary exclusion criteria.
EPA calculated T7F values for 13 invertebrate species and 30 fish species that live in
freshwater aquatic environments in North America. The data used to derive these TTF values are
provided in Appendix B. The final T7F values are listed in Table 3.10 and Table 3.11. The
presence of physiological coefficients for a taxon in Table 3.10 and Table 3.11 indicates that the
TTF values were calculated using those parameters. The absence of physiological coefficients for
a taxon indicates that EPA derived the TTF value using field data. If a TTF value could be
calculated from both physiological coefficients and field data, EPA used the TTF value
calculated from the substantially larger number of field measurements to minimize statistical
uncertainty.
Table 3.10. EPA-derived Trophic Transfer Function (TTF) Values for Freshwater Aquatic
Invertebrates.
Common name
Scientific name
AE
IR
ke
TTF
Crustaceans ^^B^
amphipod
copepod
crayfish
water flea
Hyalella azteca
copepods
Astacidae
Daphnia magna
-
0.520
-
0.406
-
0.420
-
0.210
-
0.155
-
0.116
1.22
1.41
1.46
0.74
Insects
dragonfly
damselfly
mayfly
midge
water boatman
Anisoptera
Coenagrionidae
Centroptilum triangulifer
Chironimidae
Corixidae
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
1.97
2.88
2.38
1.90
1.48
Mollusks
asian clama
zebra mussel
Corbicula fluminea
Dreissena polymorpha
0.550
0.260
0.050
0.400
0.006
0.026
4.58
4.00
Annelids
blackworm
Lumbriculus variegatus
0.165
0.067
0.009
1.29
Other
zooplankton
zooplankton
-
-
-
2.01
a Not to be confused with Corbula amurensis
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Table 3.11. EPA-Derived Trophic Transfer Function (TTF) Values for Freshwater Fish.
Common name
Scientific name
AE
IR
ke
TTF
Cypriniformes
bluehead sucker
longnose sucker
white sucker
flannelmouth sucker
common carp
creek chub
fathead minnow
sand shiner
Catostomus discobolus
Catostomus catostomus
Catostomus commersonii
Catostomus latipinnis
Cyprinus carpio
Semotilus atromaculatus
Pimephales promelas
Notropis stramineus 4
-
-
-
-
-
-
f-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
1.04
0.90
1.18
1.06
1.34
1.12
1.57
1.83
Cyprinodontiformes
mosquitofish
western mosquitofish
northern plains killifish
Gambusia sp.
Gambusia affmis
Fundulus kansae
-
-
-
-
-
-
-
-
-
0.86
1.25
1.27
Esociformes
northern pike
Esox lucius
-
-
-
2.04
Gasterosteiformes
brook stickleback
Culaea inconstans
-
-
-
1.69
Perciformes
black crappie
bluegill
green sunfish
largemouth bass
striped bass
walleye
yellow perch
Pomoxis nigromaculatus
Lepomis macrochirus
Lepomis cyanellus
Micropterus salmoides
Morone saxatilis
Sander vitreus ^^
Percaflavescens
-
-
-
-
0.375
-
-
-
-
-
-
0.335
-
-
-
-
-
-
0.085
-
-
2.67
1.48
1.27
1.27
1.48
1.82
1.42
Salmoniformes
brook trout
brown trout
mountain whitefish
cutthroat trout
rainbow trout
westslope cutthroat trout
Salvelinus fontinalis
Salmo trutta
Prosopium williamsoni
Oncorhynchus clarkii
Oncorhynchus mykiss
Oncorhynchus clarkii lewisi
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
0.88
1.44
1.38
1.07
1.19
1.20
Scorpaeniformes
mottled sculpin
sculpin
Cottus bairdi
Cottus sp.
-
-
-
-
-
-
1.38
1.29
Siluriformes
black bullhead
channel catfish
Ameiurus melas
Ictalurus punctatus
-
-
-
-
-
-
0.91
0.73
For fish species without sufficient data to directly calculate a TTF value, EPA estimated
the TTF value by sequentially considering higher taxonomic classifications until one or more
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taxa for which a calculated TTF value was available matched the tax on being considered. If the
lowest matching tax on was common to more than one species with a TTF value available, EPA
used the median TTF from the matching species. For example, although data to directly calculate
TTF for Rhinichthys atratulus (blacknose dace) were not available, this species is in the family
Cyprinidae, which also includes Cyprinus carpio (common carp), Semotilus atromaculatus
(creek chub), Pimephalespromelas (fathead minnow), and Notropis stramineus (sand shiner).
Because Cyprinidae is the lowest taxonomic classification where Rhinichthys atratulus matches
a species with an available TTF value, the median of the common carp, creek chub, fathead
minnow, and sand shiner TTF values was used as the TTF value for blacknose dace. The data
and analyses used to calculate all TTF values including those estimated by taxonomic
classification are provided in Table B-8 of Appendix B.
3.2.2.2 Derivation of Whole-Body to Egg-Ovary Conversion Factor (CF) Values
The parameter CF (conversion factor) in Equation 18 represents the species-specific
partitioning of selenium as measured in the whole-body and in egg-ovary tissue. EPA derived
species-specific CF values by applying the same method used to derive species-specific TTF
values using empirical measurements of selenium concentrations in different tissues of the same
fish. To derive whole-body to egg-ovary CF values, EPA defined matched pairs of selenium
measurements from the whole-body and from the eggs or ovaries measured from the same
individual fish or from matched composite samples. Egg-ovary concentration was defined as a
measurement from either the eggs or the ovaries. If multiple measurements from both eggs and
ovaries of the same individual or matched composite sample were available, the average value
was used. Similar to the procedure used to derive TTF values, EPA first confirmed a statistical
relationship between egg-ovary and whole body selenium for each species using OLS linear
regression of the matched pairs of measurements. If the regression resulted in a significant fit
(P<0.05) with a positive regression coefficient, EPA calculated the ratio of the egg-ovary to
whole body selenium concentration of each matched pair and used the median ratio as the CF
value for the species.
EPA had sufficient egg-ovary and whole-body selenium measurements to directly derive
egg-ovary to whole body CF values for 10 species offish. However, matched pairs of selenium
measurements in eggs and/or ovaries and muscle tissue, and matched pairs of selenium
measurements in muscle and whole body were also available. To derive CF values for additional
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fish species, EPA used either the additional data or a taxonomic classification approach to
estimate CF.
For those species offish with neither sufficient data to directly calculate an egg-ovary to
whole body CF, nor data to calculate a conversion factor for egg-ovary to muscle or whole body
to muscle, EPA first estimated CF following the approach described for the estimation of TTF
values. In this first approach, EPA sequentially considered higher taxonomic classifications until
one or more taxa for which a calculated CF value was available matched the taxon being
considered, and if the lowest matching taxon was common to more than one species with a CF
value available, EPA used the median CF from the matching species. For example, CF data are
not available to directly calculate CF ior Lepomis microlophus (redear sunfish); however, genus-
level CFs for Lepomis cyanellus (green sunfish) and Lepomis macrochirus (bluegill) are
available. Thus, EPA used the median CF values of Lepomis cyanellus and Lepomis macrochirus
for redear sunfish.
For fish species without sufficient data to directly calculate an egg-ovary to whole body
CF, but which had sufficient data to calculate a conversion factor for either egg-ovary to muscle
or whole body to muscle, EPA followed a two stage approach based on taxonomic similarity
similar to that described above. If a fish species had a species specific egg-ovary to muscle
conversion factor, but no whole body data with which to calculate an egg to whole body CF,
then available data for other species would be used to estimate a muscle to whole body
conversion factor for that species based on taxonomic relatedness. The estimated muscle to
whole body factor would be multiplied by the directly measured egg-ovary to muscle factor to
estimate an egg-ovary to whole body CF for that species. For example, rainbow trout has a
species specific egg-ovary to muscle conversion factor of 1.92, but does not have a species
specific egg-ovary to whole body CF. Using the taxonomic approach described above, the most
closely related taxa to rainbow trout with muscle to whole body conversion factors are in the
class Actinopterygii. The median conversion factor for the 8 species within that class is 1.27. The
final egg-ovary to whole body CFfor rainbow trout is 2.44 (Table 3.12), or 1.92 x 1.27.
EPA derived 10 CF values directly from matched pairs of egg-ovary and whole-body
selenium measurements and 7 CF values by multiplying EO/M and M/WB conversion factors.
Variability in the CF values for 16 of the 17 fish species was low (Table 3.12). Excluding
mountain whitefish, CFs ranged from 1.38 to 2.44, less than a 2-fold difference. CF variability
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within each species was also low for 7 of 9 species for which egg-ovary to whole-body CFs were
calculated. The two species with relatively high standard deviations for their CF values (brown
trout and cutthroat trout) contained potentially anomalous hatchery data that contributed to the
variability (see Table 3.12 footnote). These CF values are listed below in Table 3.12 and in
Table B-5 of Appendix B. All CF values including those estimated using the taxonomic
classification approach are provided in Table B-6 in Appendix B.
Table 3.12. EPA-Derived Egg-Ovary to Whole-Body Conversion Factor (CF) Values.
Common name
Scientific name
CF
Acipenseriformes
white sturgeon
Acipenser transmontanus
1.69
Cypriniformes
bluehead sucker
flannelmouth sucker
white sucker
common carp
razorback sucker
roundtail chub
Catostomus discobolus
Catostomus latipinnis
Catostomus commersonii
Cyprinus carpio
Xyrauchen texanus
Gila robusta
1.82
1.41
1.41
1.92
1.51
2.07
Esociformes
northern pike
Esox lucius
2.39
Perciformes
bluegill
green sunfish
smallmouth bass
Lepomis macrochirus
Lepomis cyanellus
Micropterus dolomieu
2.13
1.45
1.42
^^^^^ Salmoniformes
brook trout
Dolly Varden
brown trout
rainbow trout
cutthroat trout
mountain whitefish
Salvelinus fontinalis
Salvelinus malma
Salmo trutta
Oncorhynchus mykiss
Oncorhynchus clarkii
Prosopium williamsoni
1.38
1.61
1.45
2.44
1.96
7.39
Std. Dev.a
0.19
0.20
0.36
0.49
0.29
0.69
0.23
0.20
1.81b
2.03b
a Standard deviation for CF values for those species that had egg-ovary and whole body selenium
concentrations.
The brown trout and cutthroat trout standard deviations for CF values of 1.81 and 2.03 are
considerably higher than the other standard deviations in this table. The brown trout data were
taken from two studies, NewFields (2009) and Osmundson et al. (2007). CF values for three of
the four fish samples from Osmundson et al. were 4 to 6 times greater than the median. Also, the
NewFields data consisted of samples collected from natural streams and samples collected from
a fish hatchery. The CF values for the fish hatchery samples were 4 to 7 times lower than the
median value. Although collectively, the data set meets the criteria for including the brown trout
CF, the CF values for Osmundson et al. and NewFields hatchery samples may be anomalously
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high and low, respectively. Excluding these potentially anomalous data reduces the brown trout
standard deviation to 0.47. The cutthroat trout CF values are from two sources (Formation 2012
and Hardy 2005). The reason for the higher variability in the cutthroat trout CF values is due to
the relatively higher CF values in the hatchery fish from the Formation study. The standard
deviation for cutthroat trout drops to 0.62 if the hatchery fish are excluded. See Appendix B for a
presentation of the data for both of these species.
3.2.2.3 Calculation of Site-Specific Enrichment Factor (EF) Values
The most influential step in selenium bioaccumulation occurs at the base of aquatic food
webs (Chapman et al. 2010). The parameter EF characterizes this step by quantifying the
partitioning of selenium between the dissolved and particulate state. EF can vary by at least two
orders of magnitude across aquatic systems (Presser and Luoma 2010). Uncertainty in translating
a fish tissue concentration of selenium to a water column concentration using Equation 18 is
minimized when site-specific empirical observations of dissolved and particulate selenium of
sufficient quality and quantity are used to accurately characterize EF. Thus, EPA only used
aquatic sites with sufficient data to calculate a reliable EF value.
To calculate the EF of aquatic systems, EPA searched its collection of selenium
concentration measurements from field studies (see Section 2.7.7 for a description of data
sources and acceptability criteria) and identified aquatic sites with measurements from both
particulate material and the water column. EPA first identified all selenium measurements from
algae, detritus, or sediment, and then searched for corresponding water column measurements
from samples collected at the same aquatic site within one year of the particulate sample. If more
than one water measurement was available for any given particulate measurement, the median
was used. For each of these matched pairs of particulate and water measurements, EPA
calculated the ratio of particulate concentration to water concentration. If more than one ratio for
any given category of particulate material (algae, detritus, or sediment) was calculated at an
aquatic site, EPA used the median ratio. The geometric mean of the algae, detritus, and sediment
ratios was used as the site EF. Because there were at most only 3 possible values (one for algae,
one for detritus, and one for sediment), EPA used the geometric mean in order to reduce the
potential for one of the values to have excessive influence on the final site EF value.
The availability of selenium measurements from particulate material was limited. In
addition, the majority of particulate measurements were from sediment samples with a
significantly lower correlation to selenium in water (r = 0.42) compared to algae (r = 0.65; Fisher
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r-to-z transformation, P < 0.001) and detritus (r = 0.94; Fisher r-to-z transformation, P < 0.001).
Therefore, to reduce uncertainty in estimating site-specific EF values, EPA limited its analysis to
those aquatic sites with at least two particulate selenium measurements with corresponding water
column measurements, and only used sediment measurements if there was at least one other
measurement from either algae or detritus. On the basis of these requirements, EF values were
calculated for 69 individual aquatic sites.
3.2.3 Food-Web Models
For the aquatic sites with a calculated EF value, EPA modeled the food webs for the fish
species the studies indicated were present. Some of those studies provided information about the
species and proportions of organisms ingested by fish, either through direct analysis of stomach
contents, or examination of the presence and prevalence of invertebrate species. For those
studies, that site-specific information in the food web models was used. Most studies, however,
did not provide site-specific food web information. In those cases, the food web offish species
present were modeled using information about their typical diet and/or eating habits obtained
from the NatureServe database (http: //www. nature serve. org).
After EPA developed food web models, EPA identified the appropriate species-specific
TTF values for each model and calculated 777^°™?°^ Although individual TTF values were
derived for several different taxa of invertebrates and fish (Table 3.10 and Table 3.11), some of
the food web models included one or more taxa for which no TTF value was available. EPA
estimated TTF values for these taxa using the same taxonomic approach EPA used to estimate
egg-ovary to whole body, egg-ovary to muscle, and muscle to whole body conversion factors for
taxa without sufficient data. In brief, for taxa with insufficient data to calculate a TTF value,
EPA sequentially considered higher taxonomic classifications until one or more taxa for which a
TTF value was available matched the taxon being considered. If the lowest matching tax on was
common to more than one species with a TTF value available, EPA used the median TTF from
the matching species. EPA used site-specific food-web models to translate the egg-ovary
criterion element to a set of water column concentrations in order to derive the water column
concentration element of the selenium criterion. Details of these food web models are shown in
Table B-8 of Appendix B.
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3.2.4 Classifying Categories of Aquatic Systems.
Transformation reactions that convert dissolved selenium to particulate forms are the
primary route of entry into aquatic food webs, and are critical steps in selenium bioaccumulation
and toxicity (Chapman et al. 2010). Site-specific characteristics can result in substantial
bioaccumulation variability and consequently different risks of selenium toxicity for a given
dissolved selenium concentration. Freshwater systems fall into two distinct categories: lotic
systems such as rivers and streams, characterized by flowing water, and lentic systems, such as
lakes and ponds, characterized by largely standing water (e.g., Jones 1997). Water residence time
is generally shorter in lotic systems than in lentic systems, and subsequently, aquatic organisms
living in lentic systems tend to bioaccumulate more selenium than organisms living in lotic
systems for a given dissolved selenium concentration (ASTDR 2003; EPRI 2006; Luoma and
Rainbow 2005; Orr et al. 2006; Simmons and Wallschlagel 2005).
Although the distinction between lotic and lentic aquatic systems is often straightforward,
some aquatic systems possess both lotic and lentic characteristics. For example, flow rate can
vary greatly among lotic systems, with slow flowing low gradient systems (such as sloughs)
having longer residence times relative to fast flowing high gradient systems. Lotic systems can
also become more lentic during dry periods as hydrologic connectivity between deeper pools
decrease or cease with decreasing flow (Buffagni et al. 2009). Downstream reaches of some low
gradient coastal rivers can also be influenced by tides (Riedel and Sanders 1998). Some lentic
systems can exhibit some degree of flow, such as lakes fed and drained by one or more streams;
however, the magnitude of flow is generally small compared to a lotic system. Even after
accounting for flow, the majority of water movement in a lentic system is driven typically by
wind or convection rather than gravity (e.g., Jones 1997). Finally, human-made reservoirs have
some features that are intermediate between typical lotic and lentic systems. For example,
reservoirs tend to be longer and narrower than natural lakes, and they have somewhat shorter
water retention time than a natural lake of comparable volume (Thornton et al. 1990). Overall,
however, reservoirs as a general class are considered more lentic than lotic, and have historically
been classified as a type of lake (Thornton et al. 1990).
To verify the suitability of lentic and lotic aquatic system categories as the basis for
independent water column criterion values, EPA evaluated the aquatic systems that provided
data for the 69 EF values. EPA utilized the description provided by the study authors to
Do not distribute, quote or cite 76 Draft Document
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categorize each aquatic system as either lotic or lentic. Of the 27 lentic sites, the study authors
identified them as ponds (n = 16), lakes (n = 7), reservoirs (n = 3), or marshes (n = 1). Of the 42
lotic sites, the study authors identified them as creeks (n = 25), rivers (n = 6), artificial channels
(drains and wasteways, n = 4), springs (n = 2), sloughs (n = 2), or ephemeral systems (draws and
washes, n = 3). The three ephemeral aquatic sites (2 washes and 1 draw) were categorized as
lotic because there was flowing water at the time they were sampled (Butler et al. 1995; Presser
and Luoma 2009). EF values for these aquatic systems are shown in Figure 3.6.
14 -i
12 -
10 -
o) 8 •
LLJ 6 -
4 -
2 -
0
OBa
oHi
o
o
OMa
ODe
8
o
o
8
O
a 9
o
o
Q
Lake Reservoir Marsh Pond Creek Canyon Spring River Drain Draw Slough
Lentic
Lotic
Figure 3.6. Enrichment factors (EF) for 69 aquatic sites derived from published studies and
organized by waterbody type.
The dashed line represents the median EF for the 27 lentic sites (0.9 L/g), and the solid line
represents the median EF for the 42 lotic sites (0.4 L/g). See text for information on labeled
datapoints.
Because the four labeled aquatic sites in Figure 3.6 (Ba, Hi, Ma, and De) appear as
outliers, the EPA selected them for further scrutiny. Data from site "De" resulted in an EF value
of 3.3 L/g. This site was located within the tidal freshwater zone of the Delaware River (Riedel
and Sanders 1998). EPA classified this site as lotic despite the influence of tidal water because it
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77
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was well within the freshwater zone, indicating that the effects of persistent upstream
unidirectional flow outweighed tidal influence. Data from site "Ma" result in an EF value of 5.2
L/g. Site "Ma" was a small irrigation pond within the Mancos River Valley watershed in
southwestern Colorado (Butler et al. 1997). This watershed drains the Mancos Shale, a region
that is naturally high in selenium. Data from sites "Hi" and "Ba" resulted in EF values of 5.0 and
12.5 L/g, respectively. Site "Hi" was from High Rock Lake, NC, and data from site "Ba" was
from Badin Lake, NC (Lemly 1985). The high EF values at these lakes were the result of a
relatively high selenium concentration in particulate matter and low selenium concentration in
the water column.
Figure 3.7 illustrates the variability in EF values across aquatic systems and substantial
overlap between lotic and lentic categories. Some of this variability can be attributed to
differences in the ambient concentration of selenium in the water column at these aquatic sites.
EF is the ratio of selenium in particulate material (Cparticuiate) to selenium in the water column
(Cwater)- As expected, the selenium concentrations in particulate material are positively correlated
with the selenium concentrations in the water column (Figure 3.7A). The plot of Cparticuiate versus
Cwater shows a significant positive relationship for both lentic (r = 0.83, t(25) = 7.37, P < 10"6) and
lotic (r = 0.80, t(40) = 8.40, P < 10"9) aquatic systems. However, selenium accrual in particulate
matter is lower at aquatic sites with a higher water concentration of selenium compared to
aquatic sites with a lower water concentration of selenium (Figure 3.7B). The plot of Cwater
versus EF shows a significant negative relationship for both lentic (slope = -0.32, 95%
confidence interval = [-0.50, -0.15]) and lotic (slope = -0.32, 95% confidence
interval = [-0.47, -0.16]) aquatic systems. Consistent with other studies (e.g., Hamilton and
Palace 2001; Brix et al. 2005; Orr et al. 2006), these results illustrate that the overall longer
residence times of lentic systems result in increased bioaccumulation of selenium compared to
lotic systems.
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A
100 3
10 ;
Particulate
1 ,
B
10.00
1.00
*^ * • .x-*x*
f ** _*
jf .--xxx f
:.&"*' \ \
EF (L/g)
0.10
0.01
1 10
Water (ng/L)
100
0.1 1.0 10.0
Water (ug/L)
100.0
Figure 3.7. The relationship between Cwater and Cpartkuiate? and Cwater and EF for the 27
lentic and 42 lotic aquatic systems.
A: Relationship between Cwater and Cparticulate by site category.
B: Relationship between Cwater and EF by site category. Solid line, ordinary least squares linear
regression of logged data from lentic aquatic systems. Dashed line, ordinary least squares linear
regression of logged data from lotic aquatic systems.
Figure 3.8 shows the distribution of EF values grouped by lotic and lentic aquatic system
categories. Although EPA derived the lentic and lotic EF values from aquatic sites with a similar
range of water concentrations, the relative proportion of EF values collected at sites with higher
water concentrations is larger for lentic sites than lotic sites. In particular, 6 of the 27 lentic EF
values were from ponds in the Kesterson National Wildlife Refuge where Cwater ranged from
38.6 - 196 |ig/L (Saiki and Lowe 1987; Schuler et al. 1990). Despite the influence of selenium
water concentration on EF, the median of EF values from lentic and lotic aquatic systems are
significantly different from each other (Mann-Whitney U, p < 0.001). EPA concludes from these
analyses that lentic and lotic aquatic system categories are appropriate categories for
differentiating Se bioaccumulation. A listing of all aquatic-sites from which EFs were calculated
is provided in Appendix H.
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LU
6 n
5 -
4 -
3 -
2 -
1 -
o
o
Lentic (n=27) Lotic (n=42)
Waterbody Type
Figure 3.8. Distribution of EF values for the same 69 aquatic systems as shown in Figure
3.6 and 3.7 grouped by lentic and lotic aquatic system categories.
-th
-tii
th
Boxes show the 25 centile, median, and 75 centile EF values; whiskers show the 10 and 90
centiles. Circles represent EF values greater than 1.5 times the interquartile (25* -50* - lower
circles; 50* -75* -upper circles) range. Dashed line represents the median EF of all 69 sites (0.62
L/g). The EF value of 12.48 L/g from Badin Lake (Lemly 1985) is off scale.
3.2.5 Deriving Protective Water Column Concentrations for Lentic and Lotic System
Categories
To derive the water column element of the selenium criterion, EPA translated the egg-
ovary criterion element to a distribution of water column concentration values for lentic and lotic
aquatic systems. EPA uses the EF values calculated for 69 aquatic sites, available information
about the fish species present at those sites, and food web models of those fish species. Because
translation of the egg-ovary criterion element is site- and species-specific, several studies
identifying more than one species offish could potentially provide more than one translated
water column concentration (one translated water value for each species). EPA considered using
all water column values for all species present to generate distributions of translated water
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column values from lentic and lotic aquatic sites. However, the number of reported fish species
at aquatic sites with an EF value varied from one to six fish species. Furthermore, the studies
providing data for 16 of the 69 sites with EF values do not provide information on the species of
fish that may have been present at the aquatic site. Because the number offish species at an
aquatic site was not consistently reported, and because the number of reported fish species does
not necessarily indicate the number of species present at a site, EPA calculated one translated
egg-ovary criterion element to water column value for each aquatic site with both an EF value
and at least one reported fish species. When more than one species was reported at a site, the
EPA used the lowest translated water value for that site. Using this methodology, EPA translated
the egg-ovary FCV into water column concentrations at 20 lentic and 33 lotic aquatic sites. EPA
used these distributions of water concentration values translated from the egg-ovary criterion
element to derive chronic water column criterion element values for lentic and lotic aquatic
systems. Table 3.13 shows the model parameter values used to translate the egg-ovary criterion
element to site-specific water concentrations, and Figure 3.9 shows the distribution of the
translated values.
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Table 3.13. Data for the 53 Site Minimum Translations of the Egg-Ovary Criterion Concentration Element to a Water Column
Concentration.
Identification
Reference
Birkner 1978
Birkner 1978
Birkner 1978
Birkner 1978
Birkner 1978
Birkner 1978
Birkner 1978
Butler etal. 1991
Butler etal. 1993
Butler etal. 1993
Butler etal. 1995
Butler etal. 1995
Butler etal. 1995
Butler etal. 1995
Butler etal. 1995
Butler etal. 1995
Butler etal. 1995
Butler etal. 1995
Butler etal. 1995
Butler etal. 1997
Butler etal. 1997
Butler etal. 1997
Butler etal. 1997
Butler etal. 1997
Casey 2005
Casey 2005
Formation 20 12
Formation 20 12
Formation 20 12
Formation 20 12
Formation 20 12
Site
East Allen Reservoir, Medicine Bow WY
Galett Lake, Laramie WY
Larimer Highway 9 Pond, Fort Collins CO
Meeboer Lake, Laramie WY
Miller's Lake, Wellington CO
Sweltzer Lake, Delta CO
Twin Butter Reservoir, Laramie WY
Uncompahgre River at Colona
Navajo Reservoir, Piedra River Arm, near
La Boca
Spring Cr. at La Boca
Hartman Draw near mouth, at Cortez
McElmo Cr. at Hwy. 160, near Cortez
McElmo Cr. downstream from Alkali Cyn.
McElmo Cr. downstream from Yellow
Jacket Cyn.
McElmo Cr. upstream from Yellow Jacket
Cyn.
Navajo Wash near Towaoc
San Juan River at Four Comers
San Juan River at Mexican Hat Utah
Woods Cyn. Near Yellow Jacket
Cahone Canyon at Highway 666
Large pond south of G Road, southern
Mancos Valley
Mud Creek at Highway 32, near Cortez
Pond downstream from site MNP2, southern
Mancos Valley
Pond on Woods Canyon at 15 Road
Deerlick Creek
Luscar Creek
Crow Creek - 1A
Crow Creek - 3 A
Crow Creek - CC150
Crow Creek - CC350
Crow Creek - CC75
Species
Iowa darter
Iowa darter
northern plains killifish
northern plains killifish
fathead minnow
fathead minnow
fathead minnow
rainbow trout
brown trout
speckled dace
fathead minnow
speckled dace
speckled dace
fathead minnow
speckled dace
speckled dace
speckled dace
common carp
fathead minnow
green sunfish
fathead minnow
fathead minnow
smallmouth bass
fathead minnow
rainbow trout
rainbow trout
sculpin
sculpin
sculpin
sculpin
sculpin
Type
Lentic
Lentic
Lentic
Lentic
Lentic
Lentic
Lentic
Lotic
Lentic
Lotic
Lotic
Lotic
Lotic
Lotic
Lotic
7
Lotic
Lotic
Lotic
Lotic
Lotic
Lentic
Lotic
Lentic
Lentic
Lotic
Lotic
Lotic
Lotic
Lotic
Lotic
Lotic
Model Parameters
EFa
2.31
0.88
1.70
0.58
2.37
0.87
1.21
0.63
1.26
0.18
0.15
0.90
0.37
0.12
0.10
0.20
0.26
0.29
0.40
0.20
2.00
0.07
5.15
0.90
2.24
0.33
0.80
0.81
1.04
1.16
1.19
CFb
1.45
1.45
1.63
1.63
2.00
2.00
2.00
2.44
1.45
2.00
2.00
2.00
2.00
2.00
2.00
2.00
2.00
1.92
2.00
1.45
2.00
2.00
1.42
2.00
2.44
2.44
.63
.63
.63
.63
.63
ryrpi^composite-c
3.08
3.08
2.44
2.44
2.77
2.77
2.77
2.44
2.49
2.78
2.77
2.78
2.78
2.77
2.78
2.78
2.78
1.70
2.77
2.44
2.77
2.77
2.35
2.77
2.44
2.44
2.80
2.82
2.69
2.75
2.63
Translation
r d
^water
1.53
4.04
2.33
6.86
1.20
3.27
2.36
4.21
3.47
15.89
19.13
3.16
7.73
23.79
29.77
14.52
10.85
16.72
7.05
22.81
1.42
40.60
0.92
3.15
1.19
8.15
4.33
4.24
3.46
3.02
3.09
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82
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Identification
Reference
Formation 20 12
Formation 20 12
Formation 20 12
Formation 20 12
Formation 20 12
Formation 20 12
Grassoetal. 1995
Hamilton and
Buhl 2004
Lemly 1985
Lemly 1985
Lemly 1985
Saiki and Lowe
1987
Saiki and Lowe
1987
Saiki and Lowe
1987
Saiki and Lowe
1987
Saiki and Lowe
1987
Saiki and Lowe
1987
Saiki etal. 1993
Saiki etal. 1993
Saiki etal. 1993
Saiki etal. 1993
Stephens et al.
1988
Site
Deer Creek
Hoopes Spring - HS
Hoopes Spring - HS3
Sage Creek - LSV2C
Sage Creek - LSV4
South Fork Tincup Cr.
Arapahoe Wetlands Pond
lower East Mill Creek
Badin Lake
Belews Lake
High Rock Lake
Kesterson Pond 1 1
Kesterson Pond 2
Kesterson Pond 8
San Luis Drain
Volta Pond 26
Volta Wasteway
Mud Slough at Gun Club Road
Salt Slough at the San Luis National
Wildlife Refuge
San Joaquin R. above Hills Ferry Road
San Joaquin R. at Durham Ferry State
Recreation Area
Marsh 4720
Species
sculpin
sculpin
sculpin
sculpin
sculpin
sculpin
sculpin
sculpin
fathead minnow
fathead minnow
fathead minnow
western mosquitofish
western mosquitofish
western mosquitofish
western mosquitofish
western mosquitofish
western mosquitofish
bluegill
bluegill
bluegill
bluegill
J
common carp
Type
Lotic
Lotic
Lotic
Lotic
Lotic
Lotic
Lentic
Lotic
Lentic
Lentic
Lentic
Lentic
Lentic
Lentic
Lotic
Lentic
Lotic
Lotic
Lotic
Lotic
Lotic
Lentic
Model Parameters
EFa
1.55
0.24
0.54
0.45
0.69
1.32
0.86
1.32
12.48
1.75
4.99
0.51
0.32
0.60
0.36
0.93
1.03
1.37
0.43
0.36
0.75
0.10
CFb
.63
.63
.63
.63
.63
.63
2.00
1.96
2.00
2.00
2.00
1.63
1.63
1.63
1.63
1.63
1.63
2.13
2.13
2.13
2.13
1.92
r I v 1 1 iicomposite-c
2.68
3.51
2.39
2.83
2.67
2.85
2.77
2.02
2.77
2.77
2.77
2.46
2.46
2.46
2.46
2.46
2.46
2.12
2.12
2.12
2.12
1.70
Translation
r d
^ water
2.32
11.25
7.54
7.63
5.21
2.56
3.30
3.03
0.23
1.63
0.57
7.77
12.44
6.53
10.94
4.21
3.82
2.55
8.18
9.78
4.68
50.44
a - Geometric mean of the median enrichments functions (EF) for all available food types (algae, detritus, and sediment). EF (L/g) = Cfood/Cwater.
b - Taxa-specific conversion whole-body to egg ovary conversion factor (CF; dimensionless ratio).
c - Composite trophic transfer factor (TTFcomposite). Product of TTF values for all trophic levels.
d - Translated water concentration corresponding to an egg-ovary criterion element of 15.8 mg Se/kg dw, calculated by Equation 18.
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1 -
Cumulative
proportion
n
•
.
*
X
i
i
i
' •
i •
i
• *
i.
f
• X
1 1 K
1 X
• ; x
X
i
X
X
X 1
X 1
X
1
X •
X
• x
• x
X
• X
x
• x
X
X
X
X
X
X
X
X
X
X
* Lotic
• Lentic
10
100
Water (ng/L)
Figure 3.9. Probability distribution of the water column concentrations translated from the
egg-ovary criterion element at 20 lentic and 33 lotic aquatic sites.
Dashed and dash-dot lines show the 20th percentiles of the lentic and lotic distributions,
respectively.
EPA selected the 20* percentile from the distribution of translated water column values
of each category as the final national water column criterion element concentrations (3.1 |ig/L
for lotic waters and 1.2 jig/L for lentic waters) because the 20th percentile is consistent with past
practice as it provides a high probability of protection for most aquatic systems in both lentic and
lotic categories. Table 3.14 provides the 20th percentile of the water concentration values
translated from the egg-ovary criterion value.
Table 3.14. Water column criterion element concentration values translated from the egg-
ovary criterion element.
20th percentile (final EPA-recommended water
column criterion element)
Lentic
1.2 ug/L
Lotic
3.1 ug/L
As discussed in Section 2.2.2, selenium bioaccumulation potential depends on several
biogeochemical factors that characterize a particular aquatic system. Uncertainty in the
translation of the egg-ovary criterion element to site-specific water column concentrations can be
reduced by using site-specific data and information such as an EF value derived from site-
specific measurements and a food-web model derived from a biological assessment of the
aquatic system. The derivation of water column criterion element values described above is
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constrained by the need to apply a single value to a large number of aquatic systems for the
national criterion.
3.2.6 Derivation of Averaging Period for Chronic Water Criterion Element
For setting averaging periods for aquatic life criteria, U.S. EPA (1995b) used the concept
that the criterion averaging period should be less than or equal to the "characteristic time"
describing the toxic speed of action. In the context of the waterborne direct toxicity of metals,
characteristic time = 1/k, where k is the first-order kinetic coefficient in a toxicokinetic model
fitted to the relationship between LCso and exposure duration.
In the context of selenium bioaccumulation in a single trophic level, k would be the first-
order depuration coefficient, and 1/k would equal the time needed to depurate to a concentration
of 1/e times the initial concentration (where e=2.718). For depuration of two trophic levels
sequentially, invertebrates and fish, the characteristic time is likewise the time needed for c/c0 to
reach a value of 1/e.
For the first trophic level, the kinetics for algal bioaccumulation and depuration were
assumed to be rapid compared to the larger organisms at higher trophic levels; that is, the
characteristic time for algae was assumed to be negligible.
For the second trophic level, invertebrates, values for kjL2 are tabulated elsewhere in the
document. A value of O.I/day appears to be environmentally conservative, considerably higher
than those for Lumbriculus, Asian clam, and zebra mussel, but slightly lower than copepods,
which are very small in size. This corresponds to a characteristic time of 10 days.
For fish, the median depuration coefficient measured by Bertram and Brooks (1986) for
6-9 month-old (early adult) fathead minnows is applied, providing a kiL3 value of 0.02/day. This
corresponds to a characteristic time of 50 days. Because of the small size of adults of this
species, this represents faster kinetics than would likely be applicable to the salmonids and
centrarchids of greatest concern for selenium toxicity, consonant with the Newman and Mitz
(1988) inverse relationship between depuration rate and organism size. The striped bass k value
of Baines et al. (2002) is inapplicable here because it was measured in the early juvenile life
stage, a size that is too small to be relevant to reproductive impairment stemming from exposure
of adult females.
As shown in Appendix J, the characteristic time for the combined second and third
trophic levels (invertebrates and fish) is the approximate sum of the above two characteristic
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times, or 60 days. The analysis of the protectiveness of a 30-day averaging period, shorter than
the characteristic time, was performed and is shown in Appendix J. That analysis demonstrated
that a 30-day averaging period for the chronic water criterion affords protection under all
conditions, and is therefore the duration recommended for the chronic water column criterion
element.
3.3 INTERMITTENT-EXPOSURE WATER CRITERION ELEMENT: DERIVATION FROM
THE CHRONIC WATER CRITERION ELEMENT
Chapman et al. (2009) noted that selenium acute toxicity has been reported rarely in the
aquatic environment and that traditional methods for predicting effects based on direct exposure
to dissolved concentrations do not work well for selenium. As demonstrated in Appendix J, the
kinetics of selenium accumulation and depuration are sufficiently slow that attainment of the
water criterion concentration element by ambient 30-day averages will protect sensitive aquatic
life species even where concentrations exhibit a high degree of variability.
To address situations where pulsed exposures of selenium could result in
bioaccumulation in the ecosystem and potential chronic effects in fish, EPA is providing an
intermittent exposure water criterion concentration element intended to limit cumulative
exposure to selenium, derived from the chronic 30-day water criterion. To illustrate the concept,
Figure 3.10 shows a possible sequence of exposures over a 30-day period.
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o
v>
n
"c
c -
o
••••••••••••••••••••••••••I
1 10 19 28
Day
Figure 3.10. Illustration of intermittent spike exposure occurring for a certain percentage
of time (e.g., 10%) over a 30-day period, and background exposure occurring for the
remaining percentage of time (e.g., 90%).
The 30-day average concentration, Csoday, is given by:
"•"
~ Tint)
30 day = intJ
where CWis the intermittent spike concentration,/^ is the fraction of any 30-day period during
which elevated selenium concentrations occur, and Cbkgmd^ the average daily background
concentration occurring during the remaining time, integrated over 30 days. Csodayis not to
exceed the chronic criterion, WQCsoday^tho, intent is to apply a criterion element, WQdntto the
intermittent spike concentrations, then replacing CWwith WQdntand Cso day with
the above equation, and then solving for WQdnt yields:
— Cbkgrnd(l — fint~)
WQCint =
lint
The equation expresses the intermittent exposure water criterion element in terms of the
30-day average chronic water criterion element, for a lentic or lotic system, as appropriate, while
accounting for the fraction in days of any 30-day period the intermittent spikes occur and for the
concentration background occurring during the remaining time. The reasonable worst-case
assumption inherent in this approach is that selenium bioaccumulation is linear over a very wide
range of concentrations: that is, EF and T7F values do not decrease significantly as
concentrations increase.
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If the heights of three spikes in Figure 3.10 were to differ somewhat among each other,
then the intermittent criterion would apply to the arithmetic mean of the three. If the background
concentrations were to vary somewhat, then the arithmetic mean background would be entered
into the equation. Where concentrations vary smoothly over time, it does not matter where the
line is drawn defining elevated versus background concentrations. The intermittent criterion will
yield the same level of protection as the 30-day average criterion, provided that the equation uses
(a) the average of the concentrations occurring for the fraction of time defined as being
intermittently elevated, and (b) the average of the concentrations occurring for the remaining
time, defined as being background. The intermittent criterion will only be exceeded under
conditions that would have caused the 30-day criterion to be exceeded, had it been applied.
Table 3.15 illustrates example values for the intermittent water criterion concentration
element. The bottom row of the lotic and lentic values and the right column are to emphasize that
WQC/ntis not an independent criterion element but a re-expression of the 30-day average water
criterion concentration element. WQCmt converges to WQCsodayVfh&n the background
concentration is already at WQCsodayQ? when the intermittent exposure is said to occur
throughout the 30-day period.
Table 3.15.
Element.
Representative Values of the Intermittent Water Criterion Concentration
Bkgrnd
Cone,
^bkgmd
(HS/L)
0
1
2
O
3.1
0
0.5
0.7
1.0
1.2
0.033
(1 day)
Fraction of Time,/^ in a 30-day period
0.05
(1.5 days)
0.1
(3 days)
0.2
(6 days)
0.5
(15 days)
1
(30 days)
Lotic Intermittent Criterion, WQQ^ (ug/L)
93
64
35
6.0
3.1
62
43
24
5.0
3.1
31
22
13
4.0
3.1
16
12
7.5
3.5
3.1
6.2
5.2
4.2
3.2
3.1
3.1
3.1
3.1
3.1
3.1
Lentic Intermittent Criterion, WQC^ (ug/L)
36
22
16
7.0
1.2
24
15
11
5.0
1.2
12
7.5
5.7
3.0
1.2
6.0
4.0
3.2
2.0
1.2
2.4
1.9
1.7
1.4
1.2
1.2
1.2
1.2
1.2
1.2
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If the value of/Jrt, the intermittent exposure fraction of the month, is assigned a value less
than 1 day, then the intermittent criterion element value could exceed water concentrations that
have been shown to be acutely toxic to sensitive species in 2- or 4-day toxicity tests (compiled in
U.S. EPA 2004). Because the concentrations that would be acutely toxic in exposures of less
than 1 day might not be much greater than those observed to be toxic in 2-4 day exposures, the
intermittent fraction of the month must not be assigned a value less than 0.033, corresponding to
1 day.
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4 NATIONAL CRITERION FOR SELENIUM IN FRESH WATERS
The available data indicate that freshwater aquatic life would be protected from the toxic
effects of selenium by applying the following four-part criterion:
1. The concentration of selenium in the eggs or ovaries offish does not exceed 15.8 mg/kg,
dry weight; l
2. The concentration of selenium (a) in whole-body offish does not exceed 8.0 mg/kg dry
weight, or (b) in muscle tissue offish (skinless, boneless fillet) does not exceed 11.3
mg/kg dry weight; 2
3. The 30-day average concentration of selenium in water does not exceed 3.1 jig/L in lotic
(flowing) waters and 1.2 |ig/L in lentic (standing) waters more than once in three years
on average;
4. The intermittent concentration of selenium in either a lentic or lotic water, as appropriate,
does not exceed WQCint = WQC30~day " c»ftgrnd(i-/int) more than Qnce in thre£ y£ars Qn
Tint
average.3
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Table 4.1. 2014 External Peer Review Draft Freshwater Selenium Ambient Water Quality
Chronic Criterion for Aquatic Life.
Media
Type
Criterion
Element
Magnitude
Duration
Frequency
Fish Tissue
Egg/Ovary1
15.8mg/kg
Instantaneous
measurement5
Never to be
exceeded
Fish Whole
Body or
Muscle2
8.0 mg/kg
whole body
or
11.3 mg/kg
muscle
(skinless,
boneless filet)
Instantaneous
measurement5
Never to be
exceeded
Water Column3
Monthly
Average
Exposure
1.2 jig/Lin
lentic aquatic
systems
3.1 jig/Linlotic
aquatic systems
30 days
Not more than
once in three
years on
average
Intermittent Exposure4
WQCint =
WQC30_day - Cbkgrnd (1 - / int}
f int
Number of days/month with
an elevated concentration
Not more than once in three
years on average
Overrides any whole-body, muscle, or water column elements when fish egg/ovary
concentrations are measured except in certain situations. See footnote 3.
2 Overrides any water column element when both fish tissue and water concentrations are
measured except in certain situations. See footnote 3.
3 Water column values are based on dissolved total selenium (includes all oxidation states, i.e.,
selenite, selenate, organic selenium and any other forms) in water. Water column values have
primacy over fish tissue values under two circumstances: 1) "Fishless waters" (waters where
fish have been extirpated or where physical habitat and/or flow regime cannot sustain fish
populations); and 2) New (see glossary) or increased inputs of selenium from a specific source,
until equilibrium is reached.
4 Where WQCso-dayis the water column monthly element, for either a lentic or lotic system, as
appropriate. Cbkgrnd is the average background selenium concentration, and f;nt is the fraction of
any 30-day period during which elevated selenium concentrations occurs, with f;nt assigned a
value >0.033 (corresponding to 1 day). See Section 3.3.
5 Instantaneous measurement. Fish tissue data provide point measurements that reflect integrative
accumulation of selenium over time and space in the fish at a given site. Selenium concentrations
in fish tissue are expected to change only gradually over time in response to environmental
fluctuations.
EPA recommends that states and tribes adopt into their water quality standards a
selenium criterion that includes all four elements, and express the four elements as a single
criterion composed of multiple parts, in a manner that explicitly affirms the primacy of the
whole-body or muscle elements over the water column element, and the egg-ovary element over
any other element. The magnitude of the fish egg-ovary element is derived from analysis of the
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available toxicity data. The magnitudes of the fish whole-body element and fish muscle elements
are derived from the egg-ovary element coupled with data on concentration ratios among tissues.
The magnitudes of the water column elements are derived from the egg-ovary element coupled
with bioaccumulation considerations. Inclusion of the fish whole-body or fish muscle element
into the selenium criterion ensures the protection of aquatic life when fish egg or ovary tissue
measurements are not available, and inclusion of the water column elements into the selenium
criterion ensures protection when neither fish egg-ovary nor fish whole-body or muscle tissue
measurements are available, and provides consistent coverage for all waters.
To assure that the contribution of short-term exposures to the bioaccumulation risks is
accounted for in all situations, EPA is also recommending that the intermittent exposure element
be included in the selenium criterion, as noted above. EPA is not recommending a separate acute
criterion derived from the results of toxicity tests having water-only exposure because selenium
is bioaccumulative and toxicity primarily occurs through dietary exposure. Application of the
intermittent exposure criteria to single day, high exposure events will provide protection from the
most important selenium toxicity effect, reproductive toxicity, by protecting against selenium
bioaccumulation in the aquatic ecosystem resulting from short-term, high exposure events. It is
unnecessary to have an additional acute water column criterion because the intermittent exposure
criterion will be more stringent than an acute criterion. Further, as noted in this document, there
have been few if any acute exposure, water column-only selenium aquatic toxicity events
documented in the literature.
In implementing the water quality criterion for selenium under the NPDES permits
program, states may need to establish additional procedures due to the unique components of the
selenium criterion. Where states use the selenium water column concentration criterion element
values only (as opposed to using both the water column and fish tissue elements) for conducting
reasonable potential (RP) determinations and establishing water quality-based effluent
limitations (WQBELS) per 40 CFR 122.44(d), existing implementation procedures used for
other acute and chronic aquatic life protection criteria would be appropriate. However, if states
also decide to use the selenium fish tissue criterion element values for NPDES permitting
purposes, additional state WQS implementation procedures (IPs) will be needed to determine the
need for and development of WQBELs necessary to ensure attainment of the fish tissue criterion
element(s).
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The chronic selenium criterion is derived to be protective of the entire aquatic
community, including fish, amphibians, and invertebrates. Fish are the most sensitive to
selenium effects. Selenium water quality criterion elements based on fish tissue (egg-ovary,
whole body, and/or muscle) sample data override the criterion elements based on water column
Se data due to the fact, noted above, that fish tissue concentrations provide the most robust and
direct information of potential selenium effects in fish. However, because selenium
concentrations in fish tissue are a result of selenium bioaccumulation via dietary exposure there
are two specific circumstances where the fish tissue concentrations do not fully represent
potential effects on fish and the aquatic ecosystem: 1) In "fishless" waters, and 2) areas with new
or increased selenium inputs.
Fishless waters are defined as waters with insufficient instream habitat and/or flow to
support a population of any fish species on a continuing basis, or waters that once supported
populations of one or more fish species but no longer support fish (i.e., extirpation) due to
temporary or permanent changes in water quality (e.g., due to selenium pollution), flow or
instream habitat. Because of the inability to collect sufficient fish tissue to measure selenium
concentrations in fish in such waters, water column concentrations will best represent selenium
levels required to protect aquatic communities and downstream waters in such areas.
New inputs are defined as new activities (see glossary) resulting in selenium being
released into a lentic or lotic waterbody. Increased input is defined as an increased discharge of
selenium from a current activity released into a lentic or lotic waterbody. New or increased
inputs will likely result in increased selenium in the food web, likely resulting in increased
bioaccumulation of selenium in fish over a period of time until the new or increased selenium
release achieves a quasi-"steady state" balance within the food web. EPA estimates that
concentrations of selenium fish tissue will not represent a "steady state" for several months in
lotic systems, and longer time periods (e.g., 2 to 3 years) in lentic systems, dependent upon the
hydrodynamics of a given system; the location of the Se input related to the shape and internal
circulation of the waterbody, particularly in reservoirs with multiple riverine inputs; and the
particular food web. Estimates of steady state under new or increased selenium input situations
are expected to be site dependent, so local information should be used to better refine these
estimates for a particular waterbody. Thus, EPA recommends that fish tissue concentration not
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override water column concentration until these periods of time have passed in lotic and lentic
systems, respectively.
4.1 PROTECTION OF DOWNSTREAM WATERS
EPA regulations at 40 CFR 131.10(b) provide that "[i]n designating uses of a waterbody
and the appropriate criteria for those uses, the state shall take into consideration the water quality
standards of downstream waters and ensure that its water quality standards provide for the
attainment and maintenance of the water quality standards of downstream waters." Especially in
cases where downstream waters are lentic waterbody types (e.g., lakes, impoundments), or
harbor more sensitive species, a selenium criterion more stringent than that required to protect
in-stream uses may be necessary in order to ensure that water quality standards provide for the
attainment and maintenance of the water quality standards of downstream waters.
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5 SITE-SPECIFIC CRITERIA
All four elements of the selenium criterion can be modified to reflect site-specific
conditions where the scientific evidence indicates that different values will be protective of
aquatic life and provide for the attainment of designated uses.
Since the fish egg-ovary criterion element is based on toxicity data, a state may modify
that element by applying the Recalculation Procedure (U.S. EPA 2013b) to edit the species
toxicity database to reflect taxonomic relatedness to the site assemblage, while including tested
surrogates for untested resident species.
It is important to note that species in the national data set that are not present at a site
should not be deleted from the data set as those species serve as surrogate(s) for other species
known or expected to be present at a site. Confidence in the applied tissue criterion element can
be improved by further testing offish species resident at the site. The most relevant testing
would measure the survival and occurrence of deformities in offspring of wild-caught female
fish to determine an ECio for selenium in the eggs or ovaries (e.g., following Janz and
Muscatello 2008).
Using either the EPA national recommended egg-ovary, whole-body, or muscle criterion
concentration element or a site-specific egg-ovary, whole-body, or muscle criterion element,
translation of the fish tissue criterion to a water concentration can be performed in a manner that
accounts for site-specific conditions. Appendix K provides a step-wise process for deriving each
parameter used in Equation 18 to perform a site-specific translation. These steps include:
1. selecting a target fish species,
2. determining the primary food source for the target species,
3. determining the appropriate TTF values,
4. determining the appropriate EF value, and
5. determining the appropriate CF value.
Appendix K also provides information on how to obtain the site-specific information for
each step in the process. Other scientifically defensible methods, including the use of traditional
Bioaccumulation Factors (BAFs), may also be appropriate.
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Where sensitive aquatic-dependent (e.g., bird) species are known to exist, states should
consider developing site-specific criteria based on data for such species.
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6 EFFECTS CHARACTERIZATION
6.1 FISH AND AMPHIBIANS
6.1.1 Principles for Using Studies for which ECmS Cannot Be Calculated
When the data from an acceptable chronic test met the conditions for logistic regression
analysis, the ECio was used. When data did not allow calculation of ECs but did allow
calculation of closely spaced NOECs and LOECs, then the NOEC was used to approximate the
EC 10. No NOEC values were used in calculating the tissue criterion values.
When significant effects were observed at all treatment concentrations, such that no
treatment concentration was classified as a NOEC, then the chronic value was assigned as "less
than" (<) the lowest tested concentration. When no significant effects were observed at any
concentration, such that no treatment concentration was defined as an LOEC, then the chronic
value was assigned as "greater than" (>) the highest tested concentration.
A number of the chronic values in Sections 3.1.1 and 6.1.2 (reproductive effects) and in
Section 6.1.9 (nonreproductive effects) include a "greater than" (>) or "less than" (<) sign
because of an inability to resolve an exact value when all exposure concentrations of a study
yielded either too little or too much effect to provide a point estimate of a chronic value. The
decision to use chronic values with a "greater than" or "less than" sign in calculating an SMCV
followed a rule based on whether these values add relevant information to the mean, as described
below. None of these values were used in this assessment to derive the tissue criterion values.
^
6.1.1.1 Evaluation Approach
• Neither a low "greater than" value nor a high "less than" value were used to
calculate the SMCV;
• Both a low "less than" value and a high "greater than" value were included in the
SMCV calculation. However, none of these values were used in this assessment to
calculate the numeric criterion values for fish tissue.
For example, a chronic value reported here as ">15 mg Se/kg" is ignored if the tentative
SMCV is 20 mg Se/kg. The ">15 mg Se/kg" value indicates that no significant effects were
observed at the study's highest tested concentration of 15 mg Se/kg. As this is consistent with
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what would be expected if the SMCV were 20 mg Se/kg, it provides no information to support
modifying the SMCV. However, a different study showing no effects at its highest tested
concentration and yielding the value ">25 mg Se/kg" is not consistent with an SMCV of 20 mg
Se/kg, and indicates that the ">25 mg Se/kg" value provides information for modifying the mean
upwards. Conversely, a chronic value reported here as "<15 mg Se/kg" indicates that significant
effects were observed even at the study's lowest tested concentration of 15 mg Se/kg. As this is
not consistent with a 20 mg Se/kg SMCV, it indicates the utility of the "<15 mg Se/kg"
information for modifying the SMCV downwards. On the other hand, a value reported here as
"<25 mg Se/kg" would not be used to recalculate a 20 mg Se/kg SMCV. The intent of the
approach is to use all quality information that is relevant and appropriate for calculating the
SMCVs.
6.1.2 Acceptable Studies of Fish Reproductive Effects of Genera not the Four Most Sensitive
The following is a brief synopsis of the experimental design, test duration, relevant test
endpoints, and other critical information regarding the genera that were not the four most
sensitive but were included in the number of GMCVs in the dataset (see Section 3.1.5). The
studies in this section involve effects on the offspring of exposed female fish. Data are
summarized in Table 3.1. Details of these studies are contained in Appendix C.
6.1.2.1 Cyprinidae
6.1.2.1.1 Pimephales promelas (fathead minnow)
Schultz and Hermanutz (1990) examined the effects of selenium transferred from
parental fish (females) on fathead minnow larvae. The parental fathead minnows were first
exposed to selenite that was added directly to the water in artificial streams in a mesocosm study.
The selenite entered the food web and contributed to exposure via diet. Spawning platforms were
submerged into treated and control streams. The embryo samples that were collected from the
streams were brought into the laboratory and reared in incubation cups, which received stream
water dosed with sodium selenite via a proportional diluter. Edema and lordosis were observed
in approximately 25 percent of the larvae spawned and reared in natural water spiked with 10 ug
Se/L. Selenium residues in ovaries of females from the treated stream averaged 5.89 mg/kg ww
or 23.85 mg/kg dw (applying 75.3 average percent moisture for fathead minnow eggs/ovaries
from GEI Associates (2008) and Rickwood et al. (2008)).
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The reproductive SMCV for fathead minnows is <23.85 mg Se/kg dw in ovary/eggs
based on the Schultz and Hermanutz (1990) study. Because it is a "less than" value, the
sensitivity of fathead minnows relative to the species having lower SMCVs is not certain.
However, the Young et al. (2010) observation that fathead minnow populations remained after
selenium contamination of Belews Lake had eliminated most other fish species, including
bluegill and largemouth bass, indicates that fathead minnows at that site were not as sensitive as
those other species. Further support that the fathead minnow SMCV is a conservative estimate is
that it does not include the higher ECios for survival and deformities from GEI (2008), 35 - 65
mg/kg dw expressed as maternal whole body, as noted in Appendix E, Figures E-l and E-2.
6.1.2.2 Esocidae
6.1.2.2.1 Esox lucius (northern pike)
Muscatello et al. (2006) collected spawning northern pike from four sites near a uranium
milling operation in north-central Saskatoon, Canada, with egg concentrations ranging from 2.7
to 48 mg Se/kg dw. The four sites included a reference site and three sites 2, 10 and 15 km
downstream of the effluent discharge, representing a gradient of selenium exposure. Milt and ova
were stripped from gravid fish. Eggs were then fertilized in the field and incubated in the
laboratory for observations and measurements. The test was terminated when the majority of the
fry exhibited swim-up and had absorbed the yolk.
Mean egg diameter, fertilization success and cumulative embryo mortality were not
significantly different among the sites. Significant increases in percent total deformities
including edema, skeletal deformities, craniofacial deformities and fin deformities were observed
in fry originating from pike collected at the medium exposure site. The concentrations of
selenium in the northern pike eggs collected at the reference and low exposure site were very
similar, as were the percent total deformities in embryos/fry. The geometric mean of selenium in
the eggs of the adult females at the reference and low exposure sites was 3.462 mg Se/kg dw and
the corresponding arithmetic mean of the percent total deformities was 13.20%. There were only
4 adult females from exposed sites, and all had relatively similar concentrations in their eggs, all
close to the geometric mean concentration of 34.00 mg Se/kg dw. Likewise, all four exposed
females had relatively similar percent total deformities, not far from their arithmetic mean of
33.40%. This is not a sufficient level of effect for applying TRAP to determine an ECio.
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Furthermore, the relatively large spread between the two clusters of exposure concentrations
(3.462 and 34.00 mg Se/kg dw) would render a NOEC and LOEC unreliable and unsuitable for
defining a threshold. That is, the NOEC and LOEC would be "greater than" and "less than"
values, >3.462 and <34.00 mg Se/kg dw respectively, providing little information on the
sensitivity of northern pike compared to other species.
Instead, making use of the clustering of data at low exposure and effects and at elevated
exposure and effects, the effect level for the elevated exposure eggs was normalized to the low
exposure condition and rescaled to a 0-100% range. The rescaled (i.e., Abbott-adjusted) percent
of total deformities for the elevated exposure eggs was 24% (relative to the low exposure eggs).
Thus the concentration of selenium in the elevated exposure eggs (34 mg Se/kg dw) was
equivalent to an EC 24, and is the only effects concentration that can be calculated for this test,
given the limitations in the range of concentrations tested and effects observed. Although the
EC24 is not directly translatable to an ECio for use in determining the criterion, it is useful for
comparison with the EC24 in other species in order to determine species sensitivity rank. The
EC24 for skeletal deformities from the Holm et al. (2005) study of rainbow trout, calculated to be
30.9 mg Se/kg dw in eggs, is slightly lower than the northern pike value, indicating these two
species may be similar in tolerance, with the northern pike being slightly more tolerant (see
Appendix C for more details.)
6.1.2.3 Salmonidae
Seven publications provide quantitative data on the effects of selenium on salmonid
embryo/larval survival and deformity that were used in calculating criterion values. All involve
wild-caught adults taken from selenium contaminated streams and spawned for effects
determination. Exposure for all studies was therefore through the parents. Data are available for
rainbow trout (Oncorhynchus mykiss), cutthroat trout (Oncorhynchus clarkii), Dolly Varden
(Salvelinus malma) and brown trout (Salmo truttd). The studies with Oncorhynchus and
Salvelinus are discussed below; Salmo was previously discussed in Section 3.1.2.
6.1.2.3.1 Oncorhynchus mykiss (rainbow trout)
Holm (2002) and Holm et al. (2005) obtained eggs and milt from ripe rainbow trout
collected from reference streams and streams containing elevated selenium from an active coal
mine in Alberta, Canada. In 2000, 2001 and 2002 eggs were fertilized and monitored in the
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laboratory until swim-up stage, for percent fertilization, deformities (craniofacial, finfold, and
spinal malformations), edema, and mortality. No significant differences among sites were
observed for percent fertilization and mortality. Percentages of embryonic deformities and edema
were significantly different among streams, but rates of deformities at Wampus Creek, one of the
reference streams, were often similar to or higher than deformities at streams with elevated
concentrations of selenium (see Holm summary in Appendix C). The measurement of selenium
in the otolith layers of rainbow trout collected in this watershed showed low selenium exposure
in the fish's early life and a higher exposure to selenium during the fish's adult years (Palace et
al. 2007), suggesting that individuals that reach adulthood tend not to start their lives in streams
with elevated selenium concentrations, even though they may reside there later.
Estimates of effect concentrations for the combined data (i.e., 2000 through 2002) were
derived with a fitted logistic equation (TRAP). Proportion of skeletal deformities in rainbow
trout embryos was the most sensitive endpoint with an ECio of 21.1 mg Se/kg dw and an EC20 of
28.4 mg Se/kg dw. Holm (2002) and Holm et al. (2005) reported egg selenium concentrations in
wet weight. Egg wet weight was converted to dry weight assuming 61.2% moisture in rainbow
trout eggs (Seilor and Skorupa 2001).
6.1.2.3.2 Oncorhynchus clarkii lewisi (westslope cutthroat trout)
In a field study similar to those conducted by Holm et al. (2005) and Kennedy et al.
(2000), Rudolph et al. (2008) collected eggs from Westslope cutthroat trout from Clode Pond
(exposed site) and O'Rourke Lake (reference site). Clode Pond is on the property of Fording
River Coal Operations in Southeast British Columbia with reported selenium concentrations of
93 |ig/L. O'Rourke Lake is an isolated water body into which Westslope cutthroat trout were
stocked in 1985, 1989 and 1992 and has selenium levels reported as <1 |ig/L. Eggs with the four
highest Se concentrations (86.3 to 140 mg/kg dw) collected from Clode Pond fish died before
reaching the laboratory. Of those eggs from both ponds that survived, there was no correlation
between egg selenium concentration and frequency of deformity or edema in the fry. The percent
of alevins (post hatch to swim-up stage) that died was related to the selenium concentration in
the eggs; the TRAP estimates of the ECio and EC20 for survival in the eggs are 24.11 mg Se/kg
dw and 28.73 mg Se/kg dw, respectively.
As a follow-up to the study by Rudolph et al. (2008), Nautilus Environmental (2011)
conducted a more extensive study with Westslope cutthroat trout at the same site. Adult
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Westslope cutthroat trout were collected from lentic and lotic environments from locations near
the mining operations. The lentic fish were primarily captured in Clode Pond, a settling area used
to improve water quality of the mining discharge. Lotic fish were collected from the Fording
River and its tributaries near the mining operation. Reference females were obtained from
Connor Lake which is located within the watershed but not exposed to mining discharges. The
researchers reared fertilized eggs from the caught females in the laboratory until they reached
swim-up fry stage. A subset of fry surviving at swim-up was reared for an additional 28 days.
The most sensitive endpoint was larval survival at swim-up with an ECio determined by TRAP
of 24.02 mg/kg egg dw. This result is very similar to the ECio of 24.11 mg/kg egg dw
determined for the data generated by Rudolph et al. (2008). See Appendix C for more details on
the Nautilus Environmental (2011) study.
6.1.2.3.3 (Oncorhynchus clarkii bouveieri) Yellowstone cutthroat trout
Formation Environmental (2012) collected adult Yellowstone cutthroat trout from four
streams upstream and downstream of the Smokey Canyon mine in Idaho. In addition, sixteen
adults were obtained from Henry's Lake hatchery to serve as method controls. In the field,
females were stripped of eggs that were fertilized with milt stripped from males collected from
the same site. Hatchery females were stripped of eggs fertilized with milt stripped from males
obtained from the same hatchery. Fertilized eggs were placed in separate egg cups for each
female for hatching and monitoring. After swim up, remaining fry were thinned to a target of 100
fry/treatment and monitored for an additional 15 day post swim up feeding trial. Total
deformities, larval survival from hatch through 15 days post swim up, and combined deformities
r\
and survival were analyzed. Of these endpoints, the model fits for deformities (R = -0.03) and
the combined endpoint (R2=0.02) were very poor. The R2 for larval survival (0.23) TRAP model
fit was also variable, but it was included because it was both considerably less variable than the
other endpoints and because the resulting ECio of 25.25 mg/kg egg dw was consistent with other
ECioS for this species. See Appendix C for more details on the Formation Environmental (2012)
study.
Based on these three studies, for the ECio level of effects, the SMCV for cutthroat trout,
Oncorhynchus clarkii, is 24.45 mg Se/kg dw in eggs derived from Rudolph et al. (2008),
Nautilus Environmental (2011), and Formation Environmental (2012) (24.11, 24.02, and 25.25
mg Se/kg dw, respectively).
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6.1.2.3.4 Salvelinus fontinalis (brook trout)
These data were not used directly in the criterion calculations. See Section 6.1 for
discussion of the available data.
6.1.2.3.5 Salvelinus malma (Dolly Varden)
Golder (2009) collected adult Dolly Varden from a reference site and two sites
downstream from the Kemess Mine in northern British Columbia, one with a high and one with a
moderate selenium exposure in the fall of 2008. Fertilized eggs were taken to the laboratory
where they were monitored for survival and deformities until 90% of the larvae reached swim-
up, approximately 5 months after fertilization. Alevin mortality was <1% in the treatments
collected from the exposed sites and not considered an effect. The prevalence of deformities
increased sharply after the selenium egg concentration exceeded 50 mg/kg dw (Appendix C).
The proportion of Dolly Varden larvae without any type of deformity (skeletal, craniofacial, and
fmfold as well as edema), as a function of the log of the selenium concentration in the eggs using
TRAP, produced an ECio value of 56.22 mg Se/kg dw and an EC20 value of 60.12 mg Se/kg dw.
6.1.2.4 Salmonidae SMCV and GMCV Summary
For the ECio level of effects, the SMCV for cutthroat trout, Oncorhynchus clarkii, is
24.06 mg Se/kg dw in eggs derived from Rudolph et al. (2008), Nautilus Environmental (2011),
and Formation Environmental (2012) (24.11, 24.02, and 25.25 mg Se/kg dw respectively). The
GMCV for the genus Oncorhynchus is 22.71 mg Se/kg dw in eggs, derived from the 21.1 mg
Se/kg dw ECio from the combined Holm (2002) and Holm et al. (2005) rainbow trout data, the
above mean of the Rudolph et al. (2008) and Nautilus Environmental (2011) Westslope cutthroat
trout studies, and the Formation Environmental (2012) Yellowstone cutthroat trout study. The
GMCV for the genus Salvelinus is the ECio value of 56.22 mg Se/kg dw for Dolly Varden (S.
malma) from the Golder (2009) study.
6.1.2.5 Poeciliidae
Data are available for two species in this family. These studies are not represented in
Table 3.1 because they are live-bearing rather than egg-laying, but the relative tolerance of these
species is accounted for in derivation of the criterion.
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6.1.2.5.1 Gambusia holbrooki (eastern mosquitofish)
Staub et al. (2004) collected male and gravid female eastern mosquitofish from a
contaminated ash basin and a reference pond in July 1999. Male fish were used for measuring
standard metabolic rate and the reproductive endpoints. Brood size and percent viability of live
offspring at parturition were measured using the live-bearing females. Standard metabolic rates
of males, brood size of females, and offspring viability were not significantly different between
sites. Average concentrations of selenium in females were 11.85 and 0.61 mg/kg dw in the
contaminated ash basin and reference sites, respectively. The chronic value in whole body tissue
is >11.85 mg Se/kg dw whole-body (Appendix C). In a community of equally exposed fish taxa
(fish taxa having whole body tissue concentrations >11.85 mg Se/kg dw), the median egg-ovary
concentration among egg-laying fish would be expected to be 1.71 higher, or >20.26 mg Se/kg
dw.
6.1.2.5.2 Gambusia affmis (western mosquitofish)
Western mosquitofish were collected in June and July 2001 from sites in the grassland
water district in Merced County, California. Mosquitofish were collected from two sites that
were contaminated with selenium and from two reference sites in the same area with relatively
low selenium water concentrations (Saiki et al. 2004). Seventeen to 20 gravid females
(mosquitofish are live-bearers) from each site were held in the laboratory for two weeks to
quantify live and dead births and to make other measurements. Live and dead fry were visually
examined under low magnification with a binocular microscope for evidence of external
abnormalities (teratogenic symptoms such as spinal curvature, missing or deformed fins, eyes
and mouths and edema). The percentage of live births was high at both selenium-contaminated
sites (96.6 to 99.9%) and reference sites (98.8 to 99.2%). There were no obvious anomalies (e.g.,
deformities, edema) observed during the study. The concentration of selenium in 4 postpartum
females from the site with the highest selenium concentration ranged from 13.0 to 17.5 mg Se/kg
dw (geometric mean of the high and low is 15.1 mg Se/kg dw). The chronic value in whole body
tissue is >15.1 mg Se/kg dw (Appendix C). Similar to Staub et al. (2004), this value can be
converted to egg-ovary concentrations that would be expected in accompanying egg-laying fish,
by multiplying by the median fish egg-ovary to whole-body concentration ratio, 1.71. This yields
a >25.82 mg Se/kg dw equivalent egg-ovary concentration.
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Gambusia, which have been reported to be tolerant to selenium contamination, are often
one of the few remaining species at sites with high levels of selenium contamination (Cherry et
al. 1976; Lemly 1985a; Saiki et al. 2004; Young et al. 2010, Janz et al. 2010). The two studies
discussed above support this observation with a GMCV of >13.4 mg Se/kg dw in whole body
tissue, combining these "greater than" values as described in Section 6.1.1. It may be concluded
that this genus is not among the most sensitive to selenium.
6.1.2.6 Cyprinodontidae
6.1.2.6.1 Cyprinodon macularius (desert pupfish)
Besser et al. (2012), using a diet of oligochaete Lumbriculus that had fed on selenized
yeast, exposed desert pupfish to six levels of dietary and waterborne selenium. Five-week old
juveniles (Fo) were exposed for 85 days, during which time survival and growth were measured.
Upon reaching maturity at the end of this exposure period, the 60-day reproductive study was
begun, during which FI eggs were collected, counted, and further tested for percent hatch,
survival, growth, and deformities. The authors observed no significant differences in pupfish
survival, growth, total egg production, hatch, or deformities among treatments. Although the
authors noted a potential interaction between the timing of egg production and treatment, a
comprehensive re-analysis of this data, described in Appendix C, indicated that the phenomenon
was neither statistically nor biologically significant. It is concluded that the egg concentration, 27
mg Se/kg (dw), for the test's highest treatment was not sufficiently high to define a
concentration-response curve. Although desert pupfish is thus not among the most sensitive
species, the slightly reduced survival observed at 27 mg Se/kg egg dw egg suggests that the ECio
may be close to that concentration, as also noted by the authors.
6.1.3 Reproductive Effects in Catfish (Ictaluridae)
Some important families offish are not represented in the effects assessment, such as the
catfish family (Ictaluridae). In their compilation of egg-ovary versus whole-body ratios,
Osmundson et al. (2007) found comparatively high concentrations of selenium in egg-ovary
compared to whole body in black bullhead, Ameiurus melas, which are related to the Ictaluridae.
This raises a question about the potential risks of reproductive effects in this species and possibly
in related Ictaluridae. In addition to this concern about how much selenium such species may
accumulate in their eggs, U.S. Fish and Wildlife Service (2005) has suggested that offspring of
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channel catfish (Ictaluruspunctatus) and related species might be affected at unusually low egg
concentrations. This is based on results of a study in which adult female catfish were injected
with seleno-L-methionine (Doroshov et al. 1992b). Effects were found in the offspring at egg
concentrations below levels observed in other studies in Section 3.1.2 and Appendix C. These
data were not included in derivation of the criterion because the injection route of exposure is not
an acceptable experimental protocol for studies used in criterion derivation due to its difference
from exposure routes in the environment (water column and diet).
In the absence of valid tests yielding an Ictaluridae ECio or chronic value, EPA evaluated
the potential vulnerability of the taxonomic group that includes catfish by examining
comparative fisheries observations of Ictaluridae and Centrarchidae sharing the same selenium-
contaminated water body. Crutchfield (2000) reports results of annual cove rotenone sampling
performed from 1982 to 1997 in Hyco Reservoir, North Carolina. The sampling was begun after
centrarchid populations in this reservoir had collapsed due to the release of ash pond selenium
from a coal-fired power plant. The plant began operating a dry fly ash handling system in
January 1990, thereby eliminating the aquatic discharge of selenium; the sampling continued
through the recovery period.
Crutchfield (2000) reports abundance data (kg/ha) for 20 fish taxa, including four
Ictaluridae and three Centrarchidae. These data were examined to determine the relationship
between the Ictaluridae and the selenium-affected Centrarchidae populations. The correlation
matrix between annual measured abundance of the seven taxa is shown below in Table 6.1.
Correlation with the reciprocal of measured average concentrations of selenium in invertebrates
is also shown. Because the reciprocal of the selenium concentration is used, a positive correlation
means that abundance decreases as selenium concentration increases. Conversely, a negative
correlation means abundance decreases as selenium concentration decreases.
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Table 6.1. Correlation
and for selenium food
Crutchfield (2000).
matrix (values of r) for Ictaluridae and Centrarchidae abundance
chain contamination for the Hyco Reservoir data reported by
Channel catfish
White catfish
Black bullhead
Ameiurus spp.
Bluegill
Largemouth bass
Pomoxis spp.
(crappie)
1 + Inverteb.
Se Cone.
Ictaluridae
Channel White Flat Ameiurus
catfish catfish bullhead spp.
1.00 -0.36 0.18 0.68
-0.36 1.00 0.02 -0.32
0.18 0.02 1.00 0.40
0.68 -0.32 0.40 1.00
0.08 -0.31 0.32 0.22
-0.33 -0.24 -0.08 -0.24
-0.08 -0.15 0.08 -0.05
.0.44 -0.06 -0.03 -0.31
Centrarchidae
Large- Pomoxis
mouth spp.
Bluegill bass (crappie)
0.08 -0.33 -0.08
-0.31 -0.24 -0.15
0.32 -0.08 0.08
0.22 -0.24 -0.05
1.00 0.78 0.76
0.78 1.00 0.78
0.76 0.78 1.00
0.80 0.92 0.69
1 + Inverteb.
Se Cone
-0.44
-0.06
-0.03
-0.31
0.80
0.92
0.69
1.00
The centrarchid abundances are well correlated with each other and are closely related to
selenium concentrations in the food chain, with fish abundance decreasing as selenium
concentrations increase. Ictaluridae abundances, however, are unrelated either to the selenium-
sensitive centrarchid abundances or to the selenium concentrations in the food chain.
Figure 6.1 shows abundance as both mass and numbers of individuals of channel catfish
(CCF) and largemouth bass (LMB) observed by Crutchfield (2000) during the period 1982-1997.
Both species are long lived. Crutchfield (2000) notes that the decline of reproductive success and
abundance of Hyco's largemouth bass (and bluegill) was first documented in the mid-1970s.
Because this study was initiated after the largemouth bass recreational fishery had collapsed,
Figure 6.1 does not show the largemouth bass decline, only the period of its depression and
subsequent recovery.
Numbers of largemouth bass were very low at the beginning of the study period; their
numbers and mass do not begin to recover until invertebrate selenium drops below 30 mg Se/kg
dw. In the later portion of the study period, 1989-1997, largemouth bass numbers and mass
increase 100-fold. These observations are fully consistent with the premise that the earlier
observations of elevated selenium concentrations had been impairing reproduction of largemouth
bass.
In contrast, the ups and downs of channel catfish numbers, mass, and size seem to vary
randomly throughout the period of study. In 1984 catfish numbers reached their third highest
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value while their average size was at its minimum: that is, there were many young individuals.
Simultaneously, largemouth bass was near its minimum for both numbers and mass. The next
year (1985) catfish numbers jumped to their maximum for the study period, and mass reached
near maximum. Such observations are easily explained if reproduction is taking place. But they
seem inexplicable under a premise that channel catfish reproduction was even more impaired
than largemouth bass reproduction, and its population merely a senescent non-reproducing
remnant of the pre-contamination population. Rather the observations indicate that //selenium
was having any effect on catfish reproduction, it was far less than on largemouth bass
reproduction and was no hindrance to rapid population increases.
Observations of selenium-contaminated Belews Lake accord with the above observations
of Hyco Reservoir. Young et al. (2010) indicate that as many as 29 resident fish species were
documented prior to contamination, but only common carp, catfish, and fathead minnows
remained after contamination. The Doroshov et al. (1992) injection study results suggesting that
channel catfish is sensitive at egg concentrations of 5 mg Se/kg dw, four-fold below the
largemouth bass Chronic Value, thus conflict with field observations. As demonstrated in the
Appendix C discussion of the Cleveland et al. (1993) toxicity tests with juvenile bluegill, the
exposure route by which selenium was accumulated can have a dramatic influence on the
potency of a given tissue concentration. That is, to accord with the Cleveland et al. (1993) data,
the whole-body ECso would be expected to be at least 4-fold higher when accumulated via diet
than when accumulated via water. For this reason, the criterion is derived only from tests using
the environmentally relevant exposure route of diet.
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1982
1984 1986 1988 1990 1992 1994 1996
CCF kg/ha
LMB kg/ha
1982
1986 1988
1990
1992
1994 1996
1982
1984
1986
1988 1990
1992
1994 1996
*>
80
60
01
in
40
-g 20
t
V
Tilapia
accidently
introduced,^
1984^, •
All treatment
processes
operational,
early 1990
Reinitiation o
tournaments
CCF and LMB
consumption
fba
19S
ad-
^^ * ^^» ^^ visories lifted 19'
"^" **x
— -Invertebrate Se Cone
1982
1984 1986
1988 1990
1992
1994 1996
Figure 6.1. Crutchfield (2000) observations of channel catfish (CCF) and largemouth bass
(LMB) in Hyco Reservoir beginning a few years after populations of largemouth bass had
been reduced by Se contamination.
(A) number of individuals/ha, (B) mass/ha, (C) mass/ha divided by number/ha, yielding average
weight per individual, and (D) invertebrate Se concentration (mg Se/kg dw), and noting other
events relevant to management of the fishery.
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6.1.4 Reproductive Effects in Amphibians (Xenopus laevis)
Masse et al. (2014) has conducted the only maternal transfer study conducted with an
amphibian under controlled laboratory conditions. The African clawed frog (Xenopus laevis) was
fed a control diet (0.73 mg/kg Se dw) and three spiked diets containing selenium concentrations
of 10.92, 30.4 and 94.2 mg/kg dw. Trophic transfer to the frog's eggs was approximately 1:1
with measured selenium concentrations in the control and three spiked diets of 1.6, 10.82, 28.13,
and 81.66 mg/kg egg dw, respectively. Deformities were assessed in 200 tadpoles per female
(1800 - 2000 tadpoles per treatment group). ECio values determined by the authors for abnormal
spinal curvature, abnormal craniofacial structure and abnormal lens structure were 57.3, 38.4,
and 34.5 mg/kg Se egg dw, respectively. The ECio value for total deformities of 24.8 mg/kg Se
egg dw is in the mid-range of ECio values for fish (see Table 3.2). Although^ laevis is a non-
native amphibian with a different reproductive strategy, their mid-range sensitivity suggests
amphibians are protected by the fish chronic criterion. Note: the information presented here was
obtained from a platform presentation made in 2015. The authors plan to submit these data for
publication in 2015 (Janz, pers. comm.).
6.1.5 Reproductive Studies Not Used in the Numeric Criterion Derivation
6.1.5.1.1 Danio rerio (zebrafish)
Two studies (Penglase et al. 2014; and Thomas and Janz 2014) have shown the zebrafish,
Danio rerio (family Cyprinidae), to be sensitive to selenium. Penglase et al. (2014) assessed the
interaction of selenium with mercury through a maternal transfer study but did have two
treatments with selenium exposures resulting in 1.17 mg/kg egg dw (control) and 6.24 mg/kg egg
dw. The higher Se egg concentration had significantly reduced embryo survival and fecundity
relative to the control, however embryo survival in the controls was low at 54%. With only one
selenium treatment exposure, the data were not amenable to TRAP analysis. Thomas and Janz
(2014) conducted a maternal transfer study using adult zebrafish that were fed a control diet and
three levels of selenomethionine, 3.7, 9.6, and 26.6 mg/kg Se dw for 90 days before breeding the
exposed fish and collecting the fertilized embryos for assessment. TRAP analysis of larval
survival and larval deformities of 2-6 days post fertilization fish produced very low ECio values.
The lowest ECio was for deformities at 7.0 mg/kg egg dw. This value is markedly lower than any
of the ECio's in the current data set. The slope of the concentration-response curve for both
deformities and larval survival was very shallow, which was different than the selenium
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responses for all other fish species for which data were available (see Figure E-5 in Appendix E).
Further, the control mortality in the experiment continuing over 160 days was high, over 40%.
This zebrafish ECio for deformities contrasts with the absence of deformities in the
related species, fathead minnow, observed by GEI (2008) at concentrations up 40 mg/kg in adult
whole body (dw) as presented in Figure E-2 in Appendix E. The GEI (2008) fathead minnow
study was not directly used for criteria derivation because the offspring survival data for Sand
Creek appeared to be confounded by multiple stressors in this industrial waterway. However, its
deformity data appear unequivocal, indicating that the fathead minnow deformity endpoint is
relatively insensitive to selenium.
Since the zebrafish is a non-native cyprinid species, EPA assessed the information
available on zebrafish sensitivity to selenium compared to the sensitivity of native cyprinid
(minnow) species across the US (Appendix E in the criteria document), including several studies
where native cyprinids were investigated in selenium-impacted waters (NAMC 2008). Data from
these studies suggest that native cyprinids are likely less sensitive to selenium than the non-
native zebrafish.
The anomalous nature of the concentration-response curve, with the very low value
coupled with field and other laboratory data suggesting that cyprinids are not particularly
sensitive to selenium was the basis for not including the zebrafish ECio in the data for deriving
the criterion. A detailed write up of this study and a summary of field and laboratory studies
indicating native cyprinids are not one of the more sensitive families are provided in Appendix
E.
^
6.1.5.1.2 Oncorhynchus clarkii (cutthroat trout)
Kennedy et al. (2000) reported no significant differences in mortality and deformity in
eggs, larvae, and fry from wild-caught cutthroat trout between a reference and an exposed site
(Fording River, British Columbia, Canada). The observations were made on eggs reared in well
water from spawning age females collected from the two locations (N = 17 and 20, respectively)
and fertilized by one male collected at each site. The mean selenium content in eggs from fish
collected from the reference site was 4.6 mg/kg dw and from fish collected from the Fording
River was 21.2 mg/kg dw. The chronic value for eggs is >21.2 mg Se/kg dw. These values were
not used in the criterion derivation because they represent high "greater than" values, as
discussed above, and provide no additional important quantitative data for the analyses.
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Hardy (2005) fed cutthroat trout experimental diets containing a range of
selenomethionine (0-10 mg/kg dw) for 124 weeks. No significant growth or survival effects were
observed in the adult fish over the 124 weeks. The whole body concentration reached 12.5 mg/kg
dw selenium after 44 weeks. Embryo-larval observations (percent hatch and percent deformed)
were not related to whole body selenium concentrations in the spawning females (9.37 mg/kg
dw) fed the selenium-laden diet for 124 weeks. The concentration of selenium in eggs from these
females was 16.04 mg/kg dw. For this study the chronic value, an unbounded NOEC, is thus
>16.04 mg Se/kg dw in eggs. This value was not used in the criterion derivation.
6.1.5.1.3 Salvelinus fontinalis (brook trout)
Holm et al. (2005) collected spawning brook trout from streams with elevated selenium
contaminated by coal mining activity and from reference streams in 2000, 2001 and 2002.
Similar to procedures described by these authors for rainbow trout, above, fertilized eggs were
monitored in the laboratory for percent fertilization, deformities (craniofacial, fmfold, and spinal
malformations), edema, and mortality. Embryos from the contaminated stream had on average a
higher frequency of craniofacial deformities than fry from the reference stream (7.9% for the
contaminated stream compared to 2.1% in the reference stream). Although this increased rate of
craniofacial deformities was calculated to be statistically significant when compared across sites,
the Abbott-adjusted effect is only 6% and is thus below the 10% effect represented by an ECio.
But more important, when comparing across adult females (the more reliable analysis for
selenium reproductive toxicity studies of this type, and the one used to obtain the related rainbow
trout ECio for these authors' studies), there is no apparent relationship between brook trout
craniofacial deformities and exposure across a broad range of concentrations, as illustrated in
Appendix C. An environmentally conservative estimate of the NOEC might be considered to be
the average concentration of selenium in eggs from the high exposure site (Luscar Creek), >7.78
mg Se/kg ww or >20.5 mg Se/kg dw using the 61.2% moisture content for rainbow trout eggs
cited above. However, the effect threshold appears to be substantially higher based on the
absence of any consistent concentration-response relationship up to the maximum observed egg
concentration of 18.9 mg Se/kg ww or 48.7 mg Se/kg dw, as shown in the Appendix C graphs.
Given the point estimate ECio available for the related species, Salvelinus malma (Dolly Varden,
Section 6.1.2), the "greater than" chronic value for brook trout is not used to obtain the GMCV,
in accordance with the principles of Section 6.1.1.
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6.1.5.1.4 Lepomis macrochirus (bluegill)
Applicable chronic reproductive data for bluegill can be grouped by exposure type: field
and laboratory. In some field studies, chronic value estimates were "less than" fairly high
selenium concentrations (Bryson et al. 1984, 1985a; Gillespie and Baumann 1986). This low
resolution is due to the observed effect occurring at a single observed high exposure
concentration relative to a reference condition. In the Bryson et al. (1984, 1985a) and Gillespie
and Baumann (1986) studies, the artificially crossed progeny of females collected from a
selenium contaminated reservoir (Hyco Reservoir, Person County, NC) did not survive to swim-
up stage, irrespective of the origin of milt used for fertilization. Measured waterborne selenium
concentrations prior to the experiments ranged from 35 to 80 |ig/L. The ovary tissue selenium
concentration associated with this high occurrence of mortality of hatched larvae was <30 mg/kg
dw tissue, as reported by Bryson et al. (1985a), and <46.30 mg/kg dw tissue, as reported by
Gillespie and Baumann (1986). In the case of the latter, nearly all swim-up larvae from the Hyco
Reservoir females were edematous, none of which survived to swim-up.
Bryson et al. (1985b) examined percent hatch and percent swim-up larvae from spawns
using fish collected from Hyco Reservoir and a control site. There were no differences in the
Hyco measurements relative to the control. The concentration of selenium in the liver of the
parental Hyco bluegill was 18.6 mg/kg dw. The chronic values for this embryo-larval
development test was >18.6 mg Se/kg dw liver. The high "less than" and low "greater than"
chronic values obtained from Bryson et al. (1984, 1985a, 1985b) and Gillespie and Baumann
(1986) were not used in the SMCV calculation because these values are consistent with and yet
provide no numeric basis for modifying the SMCV obtained from the
6.1.6 Salmo GMCV: EPA Re-analysis of a Key Study Used in Criterion Derivation
In the draft selenium criterion document submitted for external peer review in May 2014,
the lowest GMCV in the reproductive effects dataset was for Salmo. Because of the importance
of this data for the numeric criterion calculation, and because of several experimental factors
(described below) that resulted in the calculation of several reasonable ECios, EPA conducted a
careful and thorough reanalysis of the study data and subjected its reanalysis to independent,
external peer review (ERG 2012), to confirm the validity and scientific robustness of the
approach taken by EPA in the reanalysis and use of the reanalyzed data. Those assessments were
then superseded by a reanalysis of a more complete enumeration of the deformity counts
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provided by AECOM (2012). The results of the reanalysis following the information provided by
AECOM (2012) were presented in the May 2014 draft document. During the subsequent review
phase, issues were raised about the optimal model fit for the worst case and optimistic deformity
endpoints, and about the appropriateness of assumptions made to address the accidental loss of
larval fish during the 15-day post swim up portion of the test. Additional analysis was performed
to address these issues and is reported in this document.
As described in detail in Appendix C, Formation Environmental (2011) evaluated
survival and deformities in the offspring of wild-caught brown trout having a range of exposure
levels. The evaluation of offspring had two key phases, hatch to swim-up, and swim-up to 15-
days post-swim-up. ECios were calculated for three endpoints, larval survival, larval deformities,
and combined larval survival and deformities from hatch to 15-days post swim-up. During the
second phase of the test, several drainpipe filters became clogged with uneaten food, causing
overflow loss of some organisms from several aquaria. For each endpoint, ECios were calculated
for two scenarios that examined two hypotheses regarding the condition of the fish lost to the lab
accident. In the "worst case" scenario, all fish lost to overflow were treated as being either dead
or deformed, depending on the endpoint that was evaluated. In the "optimistic" scenario, the
overflow event was treated as a random technician error unrelated to selenium toxicity, and any
lost fish were removed from the calculation. In all, a total of 6 ECios were presented in the May
2014 draft document.
The ECios presented in the May 2014 document ranged from 15.91-21.16 mg/kg. The
ECio of 15.91 mg/kg for the worst case scenario deformity endpoint was selected as the ECio for
brown trout.
During the review period, several commenters noted that the final EC 10s for the
deformity endpoint, under both the worst case and optimistic scenarios, were sensitive to initial
model conditions, in particular the parameter for the slope of the falling limb of the
concentration-response curve. For the deformity endpoint - worst case scenario, the final ECio
converged to either 15.91 mg/kg or 21.58 mg/kg depending on the initial value entered for the
slope parameter. For the deformity endpoint - optimistic scenario, the final ECio converged to
either 16.36 mg/kg, 18.37 mg/kg, or 21.94 mg/kg depending on the initial value entered for the
slope parameter. In order to evaluate the most appropriate ECio for the deformity endpoints,
models were evaluated based on residual sum of squares, and the ECio for the model with the
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lowest residual sum of squares was selected as the most appropriate. For the worst case scenario
deformity endpoint, the model with the lowest residual sum of squares was the ECio=21.58
mg/kg model, and for the optimistic deformity endpoint, the model with the lowest residual sum
of squares was the ECio=21.94 mg/kg model. This evaluation is described in detail in the brown
trout chronic summary in Appendix C. A major reason for the presence of multiple minima in
the deformity data sets was the high variability in deformities at selenium concentrations at or
below 20.5 mg/kg, including larvae hatched from hatchery fish that were fertilized by Formation
Environmental (2011) following field methodologies.
In contrast to deformities, the ECioS for hatch through 15-day post swim up larval
survival were stable across a wide range of initial model parameters. The ECio for the worst case
survival scenario was 16.78 mg/kg, and the ECio for the optimistic survival scenario was 20.40
mg/kg. Although the survival endpoint was not subject to the multiple minima issue that
complicated the ECio calculation for the deformity endpoint, the magnitude of the difference in
ECioS for the two survival scenarios, combined with the high survival among larvae that were not
lost to overflow up to 20.50 mg/kg led some commenters to question the validity of the worst
case assumption that all larvae lost to overflow were dead or deformed.
Because of the uncertainty in how best to address the issue of larval fish lost to overflow
during the second portion of the test, an ECio was calculated for survival for the first portion
(hatch to swim up) of the test. The first portion of the test was much longer than the second
portion of the test (88 days on average compared to 15 days), and by omitting the second portion
of the test, the analysis is free from all assumptions regarding how to address the uncertainties
surrounding the lab accident. For the calculation of this endpoint, larvae that survived to the end
of the first portion of the test but did not reach swim up were included as surviving. In contrast,
these larvae were assumed to have died when hatch through post swim up survival was
calculated. Although counting all surviving larvae as survivors is less conservative in theory, the
resulting ECio is lower (18.09 mg/kg with all survivors counted vs. 20.62 mg/kg with survivors
who did not swim up assumed to be dead). Only survival could be assessed for the first portion
of the test, because visibly non-deformed fish were preferentially selected for the post swim up
portion of the test during the thinning process. The final recommended ECio for brown trout in
this version of the draft selenium criterion is 18.09 mg/kg for survival for the first portion of the
test (hatch to swim up).
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6.1.7 Impact of Number of Tested Species on Criterion Derivation
Many of the species used for testing the toxicity of selenium are those observed to be
affected at contaminated sites or otherwise suspected to be particularly sensitive. Six of the 8
minimum data requirements were met, and the other 2 (for planktonic and benthic crustaceans)
were waived (see Section 2.6). Of the N=15 genera used for the calculation of the criterion, ten
are fish, which are more sensitive than invertebrates, based on the available data. Of the ten fish
genera, five are either salmonids or centrarchids. Had a broader array of expected insensitive
taxa been included, then the four most sensitive genera would not likely change, but N would
increase. The criterion calculation for selenium is relatively insensitive to the effect of increasing
the value of N by adding more tests with different genera than those already represented. Setting
N=20 (leaving the four most sensitive the same) would only raise the criterion from 15.8 mg
Se/kg to 16.2 mg Se/kg. This insensitivity occurs because the four lowest GMCVs are closely
spaced, such that the calculated egg-ovary criterion element is never distant from the lowest
GMCV.
6.1.8 Comparisons between Concentrations in Different Tissues
Researchers often report concentrations of selenium in fish eggs or ovaries (e.g.,
Formation Environmental 2011; Formation Environmental 2012; Holm et al. 2005; Osmundson
et al. 2007). Osmundson et al. (2007) found reduced levels of selenium in ovaries after
spawning, presumably due to the loss of selenium through spawning and release of eggs with
relatively high concentrations of selenium. Of the 14 chronic values determined from the
maternal transfer reproductive studies, 11 values represent selenium measured in eggs. Three
values represent selenium measured in the ovaries: Schultz and Hermanutz (1990), Hermanutz et
al. (1992, 1996) and Carolina Power & Light (1997). However, information in two of these
studies indicates that the concentrations of selenium in the ovaries were similar to concentrations
in eggs. Schultz and Hermanutz (1990) measured selenium in fathead minnow ovaries at the end
of the study from fish that presumably had spawned. The authors found the concentrations of
selenium in the ovaries and embryos of the fathead minnows exposed to the same treatments to
be similar. Hermanutz et al. (1992, 1996) sampled adult female bluegill just prior to spawning
and at the end of the test (post spawning) and found no decreases in the concentration of
selenium in the post-spawned fish. In the third study, selenium in ovaries of largemouth bass
(Carolina Power & Light 1997) was measured from fish sampled just after spawning. Based on
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the indications that the selenium concentrations in the ovaries were similar to that in eggs in the
Schultz and Hermanutz (1990) and Hermanutz et al (1992, 1996) studies, egg selenium and
ovary selenium were considered equal for the toxicity data set. Any potential error resulting from
this assumption would be conservative since the effect of spawning only lowers the selenium
concentration in the ovary. EPA recognizes selenium ovary concentrations may vary in field
collected samples due to fish reproductive cycles and will address such concerns in the
implementation information.
6.1.9 Studies of Non-Reproductive Effects
Non-reproductive effect studies do not involve effects on the offspring of exposed female
adults, and their results are not expressed as selenium concentrations in egg or ovary tissue.
Because selenium concentrations in whole body and muscle are generally lower than in egg and
ovary, with observed egg-ovary to whole-body ratios ranging from 1.3 to 7.4, and egg-ovary to
muscle ratios ranging from 1.0 to 5.8, whole-body and muscle effect concentrations cannot be
directly compared to egg-ovary effect concentrations. Non-reproductive effects were determined
to provide a less reliable basis for a criterion, in part because comparatively few of such studies
provided sigmoidal concentration-response curves. Non-reproductive SMCVs and GMCVs are
shown in Table 6.2 below and summaries of the acceptable non-reproductive studies are
included in Appendix D.
6.1.10 Special conditions for consideration of primacy of water column criterion elements over
fish tissue criterion elements
1 The chronic selenium criterion is derived to be protective of the entire aquatic community,
including fish, amphibians, and invertebrates. Fish are the most sensitive to selenium effects.
Selenium water quality criterion elements based on fish tissue (egg-ovary, whole body,
and/or muscle) sample data override the criterion elements based on water column Se data
due to the fact, noted above, that fish tissue concentrations provide the most robust and
direct information on potential selenium effects in fish. However, because selenium
concentrations in fish tissue are a result of selenium bioaccumulation via dietary exposure,
there are two specific circumstances where the fish tissue concentrations do not fully
represent potential effects on fish and the aquatic ecosystem: 1) In "fishless" waters, and 2)
new or increased selenium inputs.
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Fishless waters are defined as waters with insufficient instream habitat and/or flow to
support a population of any fish species on a continuing basis, or waters that once supported
populations of one or more fish species but no longer support fish (i.e., extirpation) due to
temporary or permanent changes in water quality (e.g., due to selenium pollution), flow or
instream habitat. Because of the inability to collect sufficient fish tissue to measure selenium
concentrations in fish in such waters, water column concentrations will best represent
selenium levels required to protect aquatic communities and downstream waters in such
areas.
New inputs are defined as new activities (see glossary) resulting in selenium being released
into a lentic or lotic waterbody. Increased input is defined as an increased discharge of
selenium from a current activity released into a lentic or lotic waterbody. New or increased
inputs will likely result in increased selenium in the food web, likely resulting in increased
bioaccumulation of selenium in fish over a period of time until the new or increased
selenium release achieves a quasi-"steady state" balance within the food web. EPA estimates
that concentrations of selenium fish tissue will not represent a "steady state" for several
months in lotic systems, and longer time periods (e.g., 2 to 3 years) in lentic systems,
dependent upon the hydrodynamics of a given system; the location of the Se input related to
the shape and internal circulation of the waterbody, particularly in reservoirs with multiple
riverine inputs; and the particular food web. Estimates of time to achieve steady state under
new or increased selenium input situations are expected to be site dependent, so local
information should be used to better refine these estimates for a particular waterbody. Thus,
EPA recommends that fish tissue concentration not override water column concentration
until these periods of time have passed in lotic and lentic systems, respectively.
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Table 6.2. Freshwater Chronic Values from Acce
Species
Acipenser
transmontanus
white sturgeon
Pogonichthys
macrolepidotus
Sacramento splittail
Pimephales promelas
fathead minnow
Pimephales promelas
fathead minnow
Xyrauchen texanus
razorback sucker
Xyrauchen texanus
razorback sucker
Catostomus latipinnis
flannelmouth sucker
Oncorhynchus
tshawytscha
Reference
Tashjian et al.
2006
Teh etal. 2004
Bennett etal. 1986
Dobbsetal. 1996
Beyers and
Sodegren2001a
Beyers and
Sodegren2001b
Beyers and
Sodegren2001a
Hamilton et al.
1990
Exposure route and
duration
dietary (lab)
8 weeks
dietary (lab)
9 months
dietary (lab)
9 to 19 days
dietary and
waterborne
(lab)
8 days
dietary and
waterborne (lab)
28 days
dietary and
waterborne (lab)
28 days
dietary and
waterborne (lab)
28 days
dietary (lab)
60 days
ptable Tests - Non-Reproductive Endpoints (Parental Females Not Exposed).
Selenium form
seleno-L-methionine in
artificial diet
seleno-L-methionine in
artificial diet
selenized-yeast
algae exposed to selenite
then fed to rotifers which
were fed to fish
algae exposed to selenate
in water then fed to
rotifers which were fed
to fish
water: selenate; diet:
algae exposed to selenate
in water then fed to
rotifers which were fed
to fish
water: site waters; diet:
algae exposed to site
water then fed to rotifers
which were fed to fish
water: selenate;
diet: algae exposed to
selenate in water then
fed to rotifers which
were fed to fish
mosquitofish spiked with
seleno -DL-methionine
Toxicological
endpoint
ECio juvenile
growth
EC2o juvenile
growth
NOEC
LOEC
MATC juvenile
deformities
(juvenile exposure
only)
Chronic value for
larval growth
LOEC for larval
fish dry weight after
8d
NOEC for survival
and growth
NOEC for survival
and growth
NOEC for survival
and growth
ECio for juvenile
growth
Chronic value,
mg/kg dwa
15.08 WB
27.76 M
17.82 WB
32.53 M
10.1 M
15. 1M
12.34 M
51.40 WB
<73 WBb
>12.9WBb
>42 WBb
>10.2 WB
7.355 WB
SMCV
mg/kg dw
EC10
15.1 WB
27.8 M
EC20
17.8 WB
32.5 M
10.1 M
15. 1M
12.3 M
51.40WB
69.83 M
see text
>10.2WB
EC10
9.052 WB
GMCV
mg/kg dw
15.1 WB
27.8 M
10.1 M
15. 1M
12.3 M
51.40WB
69.83 M
see text
>10.2WB
EC10
9.052 WB
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Species
chinook salmon
Oncorhynchus mykiss
rainbow trout
Oncorhynchus mykiss
rainbow trout
Morone saxitilis
striped bass
Lepomis macrochirus
bluegill
Lepomis macrochirus
bluegill
Reference
Hilton and Hodson
1983;
Hicks etal. 1984
Hilton etal. 1980
Coughlan and
Velte 1989
Lemly 1993a
Mclntyre et al.
2008
Exposure route and
duration
dietary (lab)
16 weeks
dietary (lab)
20 weeks
dietary (lab)
80 days
dietary and
waterborne (lab)
180 days
20 to 4°C
dietary and
waterborne (lab)
180 days 20°C
dietary and
waterborne (lab)
182 days
20to4°C(ESl)
dietary and
waterborne (lab)
182 days
20 to 9°C (ESS)
dietary and
waterborne (lab)
182 days
20 to 4°C (ES2)
Selenium form
mosquitofish spiked with
SLD diet
sodium selenite in food
preparation
sodium selenite in food
preparation
Se-laden shiners from
Belews Lake, NC
diet: seleno-L-
methionine
water: 1:1
selenate: selenite
diet: seleno-L-
methionine
water: 1:1
selenate: selenite
diet: Lumbriculus fed
selenized-yeast
water: 1:1
selenate: selenite
diet: Lumbriculus fed
selenized-yeast
water: 1:1
selenate: selenite
diet: seleno-L-
methionine
water: 1:1
selenate: selenite
Toxicological
endpoint
EC20 for juvenile
growth
EC10 for juvenile
growth
EC20 for juvenile
growth
juvenile growth
NOEC
LOEC
MATC
juvenile survival
and growth
NOEC
LOEC
MATC
LOEC for survival
of yearling bass
LOEC for juvenile
mortality at 4oC
Threshold prior to
"winter stress"
NOEC for juvenile
mortality at 20oC
EC10juv. survival
ESI
EC20juv. survival
ESI
ECiojuv. survival
ESS
EC2ojuv. survival
ESS
NOECjuv. surv.
ES2
Chronic value,
mg/kg dwa
10.47 WB
11.14WB
15.73 WB
21 Liver
7 1.7 Liver
38.80 Liver
40 Liver
100 Liver
63. 25 Liver
<16.2 Me
<7.91 WB
5.85 WB
>6.0WB
9.27 WB
9.78 WB
14.00 WB
14.64 WB
>9.992 WB
SMCV
mg/kg dw
EC20
12.83 WB
NOAEC
28.98 L
LOAEC
84.68 L
MATC
49.52 L
<16.2M
4°C
EQo-NOAEC
8.15 WB
4°C
EC20-LOAEC
8.80 WB
9°C EC10
14.0 WB
9°C EC20
14.6 WB
GMCV
mg/kg dw
<16.2M
8.15 WB
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Species
Lepomis macrochims
bluegill
Lepomis macrochims
bluegill
Reference
Bryson et al.
1985b
Cleveland et al.
1993
Exposure route and
duration
dietary (lab)
60 days
dietary (lab)
90 days
Selenium form
seleno-DL-cysteine
seleno-L-methionine
Toxicological
endpoint
NOEC for juvenile
growth
NOEC for juvenile
survival
Chronic value,
mg/kg dwa
>3.74WBb
>13.4WBb
SMCV
mg/kg dw
GMCV
mg/kg dw
All chronic values reported in this table are based on the measured concentration of selenium in whole body (WB), muscle (M)
or liver (L) tissues.
Chronic value not used in SMCV calculation (see text).
Tissue value converted from ww to dw. See Appendix C for conversion.
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6.1.11 Comparison of Fish Chronic Reproductive Effects and Chronic Non-Reproductive
Effects
A chronic criterion element of 15.8 mg/kg dw in the egg/ovary addresses the toxic effect
identified by the Chapman et al. (2009, 2010) expert workshop to be of greatest concern, and is
expected to be protective of non-reproductive endpoints such as juvenile survival and growth.
If the information in the reproductive-effect GMCV Table 3.5 or Figure 3.2 (expressed
as whole-body) were combined with the information in the nonreproductive-effect Table 6.2 or
Figure 6.2, and the lower of the reproductive or nonreproductive GMCVs for each taxon used to
construct a combined distribution of whole-body chronic values, the resulting criterion
(corresponding to N=18, accounting for three additional fish genera only having
nonreproductive-effect GMCVs), the criterion would be calculated to be 8.14 mg Se/kg WB dw,
slightly higher than the reproductive-effect criterion expressed as whole-body (Figure 6.3).
"^ 1 TO
M 128
01 fiA
Bfl "4
£
J OT
C ^Z
3
•§ 16
00
01
O S
o 8
A .
C
« fish (TL3) WB Nonrepro GMCV
A lrwerteb(TL2)GMCV
is ^
A
A
• *
•• — . ^. . ^L .......................
) 2 4 6 8 10
Rank
Figure 6.2. Distribution of (a) fish (Trophic Level 3) non-reproductive GMCVs for fish
measured as whole-body concentrations or muscle concentrations converted to whole body,
and (b) invertebrate (Trophic Level 2) GMCVs, and (c) the WB criterion applicable to
TL3.
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g
M
Ol
l/l
J
3
>
o
00
Ol
.c
5
TITC nn
190 nn
64.00
37 nn
16.00
A nn
• Fish (TL3) WB Repro GMCV
• Fish (TL3) WB Nonrepro GMCV
A Irwerteb (TL2) GMCV
-----Fish(TL3)WBFCV
^
c
A 0
V 9
^^
0 2 4 6 8 10 12
Rank
Figure 6.3. Distribution of fish reproductive effect GMCVs from Figure 3.2 and
distribution of fish nonreproductive effect GMCVs and invertebrate GMCVs from Figure
6.2.
W
For establishing a reliable criterion, the sufficiency of and consistency among the data
underlying the reproductive-endpoint GMCVs favor their use over any non-reproductive
endpoint data (see Section 3.1.1 and Appendix C). Most of the reproductive studies involved
examining the offspring of wild-caught females, exposed under real-world conditions. Most had
unambiguous concentration-response curves that supported ECio estimates.
In contrast, the non-reproductive endpoint studies provide fewer data for supporting a
criterion, and fewer of these studies yielded the type of concentration-response data that could
support EC 10 estimates. Furthermore, the non-reproductive data are not as consistent, as noted by
Janzetal. (2010).
In contrast to the non-reproductive effects, the reproductive effects show more clear-cut
concentration-response relationships (11 of the 19 reproductive chronic values are specific ECs,
compared to only 5 of the 19 non-reproductive chronic values), are more readily reproducible,
and are better corroborated by field observations. Reproductive effects represent the endpoint of
greatest concern (Chapman et al. 2009, 2010); all non-reproductive GMCVs are protected by a
criterion derived from the reproductive GMCVs. The reproductive endpoint data, expressed
relative to selenium concentrations in fish eggs and ovaries, thus provide a more reliable and
protective basis for the criterion. Because the data set used to derive the criterion is comprised
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primarily of the aquatic species considered most sensitive to selenium (salmonids and
centrarchids) and because the criterion is designed to protect 95% of the genera, the criterion of
15.8 mg/kg dw ovary/egg should be protective of aquatic populations offish and invertebrates.
6.2 WATER
6.2.1 Validation of Translation Equation for Developing Water Column Concentrations
EPA evaluated the efficacy of the equation used to translate the egg-ovary criterion
element to a water column concentration. EPA's translation equation is given as:
ri _ ->
water r^rr^-j-^ composite T^T^ t~*T^
TTF xEFxCF (Equation 18)
Because fish bioaccumulate selenium over a relatively long time period, single measurements of
selenium in fish tissue are likely to be less variable and a better representation of selenium loads
to the aquatic system than single measurements of selenium in the water column. Thus, EPA
used a validation approach based on fish tissue measurements rather than single water
measurements.
The EPA solved Equation 18 for egg-ovary concentration yielding:
Cegg.omfy = Cwater xTTFcompos"e xEFxCF (Equation 19)
EPA used Equation 19 to calculate the predicted concentration of selenium in the eggs and
ovaries offish from all spatially and temporally relevant measurements in the water column.
EPA then compared those predicted values to the measured concentration in the fish.
EPA searched its collection of selenium measurements in fish tissue taken from aquatic
sites with a previously calculated EF value. Identified tissue measurements from other than eggs
or ovaries were converted to equivalent egg-ovary concentrations using species-specific
conversion factors as described previously. For each tissue measurement, EPA searched its
collection of selenium measurements again for water column measurements that were taken from
the same aquatic site and within one year of the tissue measurement. If more than one water
column measurement was matched to a tissue measurement, the median water column
measurement was used. For each matched pair of tissue and water measurements, appropriate
species-specific TTF and CF values were identified as described previously, and the EF value
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from the site samples were taken. EPA then used Equation 19 to calculate the predicted egg-
ovary concentration from the observed water column concentration. Finally, EPA compared the
predicted egg-ovary concentrations with the observed egg-ovary concentrations.
EPA identified 169 tissue measurements associated with one or more water column
measurements. A predicted egg-ovary concentration was calculated for each water column
concentration as described above. Figure 6.4 shows all 169 predicted egg-ovary concentrations
plotted against the measured egg-ovary concentrations. Because both the predicted and observed
selenium concentrations exhibited substantial heteroscedasticity (the variability of one variable is
unequal across the range of values of a second variable that predicts it), they are plotted and
analyzed on a log scale. The predicted and measured concentrations are highly correlated
(r=0.81, t(i67)=17.91, PO.001). Data used to generate Figure 6.4 can be found in Appendix I.
1000.0 3
Observed egg-ovary
concentration (mg/kg
dw)
100.0
10.0
1.0 --
0.1
Egg-ovary criterion element
I
Egg-ovary
criterion
element
0.1 1.0 10.0 100.0 1000.0
Predicted egg-ovary concentration (mg/kg dw)
Figure 6.4. Scatter plot of predicted versus measured concentrations of selenium in fish.
Solid line shows unity y = x line; dashed lines show the egg-ovary criterion element value.
Although there is a strong correlation between predicted and observed egg-ovary
concentration values, Figure 6.4 shows more data points above the y = x line at low selenium
concentrations. This result suggests the model underestimates bioaccumulation at low selenium
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concentrations. Such behavior is likely the result of the inherent model assumption of constant
bioaccumulation rates regardless of selenium concentration, whereas selenium bioaccumulation
has been shown to be inversely related to water concentration (see Sections 3.2.2 and 3.2.4 for
further discussion). Within the range of concentrations near the egg-ovary criterion element
value, however, the relationship between predicted and observed selenium concentrations are
evenly dispersed around the y = x line. Thus the model is unlikely to result in biased estimates
near egg-ovary concentrations that may require regulatory action.
Dispersion around the unity line is likely attributable to several sources of uncertainty
including small sample sizes, temporal or spatial variability in selenium exposure, and local
variability in aquatic food webs. EPA limited this analysis to only those aquatic sites with at least
two paniculate measurements available to calculate an EF value and with at least one of them
from algae or detritus. Nevertheless, only one or two measurements of algae and/or detritus were
available for 41 of the 69 aquatic sites evaluated. Although such a restriction reduces uncertainty
when applying Equation 19 to available data, EPA believes that two paniculate measurements
are only marginally sufficient. Another potential source of uncertainty is the frequent absence of
site-specific information about the types and proportions of organisms ingested by fish. In most
cases, EPA estimated the type and proportion of prey organisms using general knowledge of the
fish species and aquatic system location. Notwithstanding the limitations in available data, the
EPA concludes from this analysis that Equation 18 provides a reasonable translation of the egg-
ovary criterion to a site-specific water concentration.
6.3 PROTECTION OF THREATENED OR END ANGERED SPECIES
The chronic toxicity dataset for selenium contains toxicity data for two Federally-listed
endangered species, Cyprinodon macularius (desert pupfish) and Oncorhynchus mykiss (listed as
steelhead, indicating anadromous individuals, but herein called rainbow trout, implying non-
anadromous individuals). The dataset also contains toxicity data for Acipenser transmontanus
(white sturgeon) which is listed as endangered in specific locations, such as the Kootenai River
white sturgeon in Idaho and Montana. The white sturgeon also serves as a surrogate for other
sturgeon listed as threatened or endangered (e.g., pallid and shovelnose sturgeon). The Acipenser
GMCV of 16.3 mg/kg dw egg is the lowest value in the dataset and therefore provides protection
for other potentially sensitive sturgeon.
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Desert pupfish, Cyprinodon macularius, with a chronic value estimated to be >27 mg
Se/kg dw egg, is not among the most sensitive species. Its chronic value of >27 mg Se/kg dw egg
is substantially above the chronic egg-ovary criterion element of 15.8 mg Se/kg dw.
Oncorhynchus mykiss has a SMCV of 21.1 mg Se/kg dw egg, and is in the fourth most
sensitive genus. The dataset contains multiple studies with cutthroat trout (Oncorhynchus clarkii)
some subspecies of which are Federally listed as threatened. The SMCV for cutthroat trout is
24.06 mg Se/kg dw egg. Both of these chronic values for Oncorhynchus species are greater than
the chronic egg-ovary criterion element.
The dataset also contains toxicity information for Salvelinus malma (Dolly Varden)
which is not threatened or endangered, but is so closely related to the threatened Salvelinus
conjluentus (bull trout) that it can hybridize with that species, producing fertile offspring (Baxter
et al. 1997). Dolly Varden is the least sensitive fish species for which information is available,
with SMCV of 56 mg Se/kg dw egg. Salvelinus fontinalis, brook trout, can also hybridize with
bull trout, but the offspring are sterile, suggesting that it is less closely related. With the available
study of brook trout, although Section 6.1.2 conservatively sets the NOEC at >20.5 mg Se/kg dw
egg, which was the average concentration at the Holm et al. (2005) high-exposure site, the
concentration-response information for the offspring of individual females, presented in
Appendix C, suggests that its ECio could be substantially higher, possibly as high as that for
Dolly Varden.
The criterion of 15.8 mg Se/kg (dw) egg-ovary element is below all of the above
mentioned chronic values for threatened and endangered (or closely related) species. However,
because other threatened or endangered species might be more sensitive, if relevant new
information becomes available in the future, it should be considered in state- or site-specific
criteria calculations.
The protectiveness of the draft whole body criterion of 8 mg/kg dw to threatened and
endangered species is supported by a recent non-reproductive study with two sturgeon species.
De Riu et al. (2014) fed juvenile green and white sturgeon (-30 g body weight) diets containing
a range of selenium concentrations (selenomethionine added to diet formulation; 2.2 mg/kg Se in
control diet (no added Se) and 19.7, 40.1 and 77.7 mg/kg Se in the three treatment diets). Several
endpoints were monitored over the 8 week exposure period including survival and percent body
weight increase (% BWI). White sturgeon had no mortalities through the highest dietary
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treatment. Green sturgeon juveniles had 0%, 7.7% and 23.1% mortality with the three dietary
treatments. TRAP analysis (threshold sigmoid nonlinear regression) of the green sturgeon
survival data resulted in a whole body ECio value of 28.93 mg/kg dw. ECio values were lower
for % BWI using TRAP. For % BWI, the whole body ECio value for green sturgeon was 16.36
mg/kg dw, and 23.94 mg/kg dw for white sturgeon.
Also notable, the background concentrations of selenium in the juvenile green and white
sturgeon were also elevated at 7.2, 6.5 and 7.1 mg/kg dw (green sturgeon whole body), and 4.8
7.3 and 5.6 mg/kg dw (white sturgeon whole body) at test initiation, and after 4 and 8 weeks of
exposure, respectively.
The De Riu et al. (2014) study suggests that green sturgeon may be more sensitive to
selenium than white sturgeon and also that the draft EPA whole body concentration of 8.0 mg/kg
dw will be protective, based on the survival and growth data and the observation that the control
whole body tissue concentrations are similar to the proposed criterion. This is important because
white sturgeon, as well as juvenile green sturgeon (up to 3 to 4 yrs), spend most of their time in
the coastal rivers and estuaries. All species in the Acipenseriforms (sturgeon and paddlefish)
spawn in freshwaters (Bemis and Kynard 1997) or spend their entire life in freshwater. The
inclusion of the white sturgeon's ECio in the dataset provides surrogacy for the threatened and
endangered species from this group.
6.3.1 Special Consideration for Pacific Salmonid Juveniles
The current draft criterion is based on reproductive effects (larval mortality and/or
deformities) for offspring of selenium-exposed adults, and the whole-body criterion element is
derived from the egg-ovary element, with an implicit assumption of adult exposure to selenium.
One peer-reviewer of the EPA External Peer Review Draft criterion document raised concerns
regarding the protection of anadromous salmonids, since there is at least some evidence (e.g.,
Hamilton et al. 1990) that juvenile growth may be comparable in sensitivity to reproductive
effects endpoints used by EPA. Anadromous salmon species (e.g. Chinook salmon) in the Pacific
Northwest are unique in that reproductively mature adults are not exposed to selenium in the
freshwater environment due to their life history (i.e., young juvenile salmon leave freshwater
streams and rivers as smolts and mature to adulthood in the marine environment until migration
for spawning begins. Furthermore, they are semelparous, so there is no potential Se exposure
following spawning in freshwater.).
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Juvenile salmon have evolved different strategies for growth and maturation to the smolt
stage, and may spend from 3 months to 2 years in freshwater (depending on timing of egg
hatching and other factors) before migrating to estuarine areas as smolts and then into the ocean
to feed and mature. Salmon remain in the ocean for 1 to 6 years (more commonly 2 to 4 years),
with the exception of a small proportion of yearling males (called jack salmon), which mature in
freshwater or return after 2 or 3 months in salt water (NOAA 2011).
The physiological and morphological changes that allow these species to adapt to marine
conditions as juveniles are reversed in returning adults preparing to migrate up natal streams to
spawn. One key change is the cessation of feeding prior to re-entry into freshwater. Since mature
females are not feeding after returning to freshwater, it is not representative to predict
reproductive effects for anadromous salmonid species based on egg-ovary selenium
concentrations, because the exposure is wholly from selenium sources in the marine environment
(Groot and Margolis 1991).
6.3.1.1 Selenium Toxicity to Juvenile Salmonids
Hamilton et al. (1990) assessed the toxicity of two organoselenium diets in 90-day partial
life cycle tests in freshwater with two life stages of Chinook salmon (Oncorhynchus
tshawytscha). The first diet consisted offish meal made from low-selenium mosquitofish
(collected from a reference site) fortified with selenomethionine (here termed the SeMet diet).
The second diet contained fish meal made from high-selenium mosquitofish (Gambusia a/finis)
collected from the San Luis Drain (SLD), California (here termed the SLD diet). This waterbody
is known to have high concentrations of selenium. A 90-day partial life cycle study was
conducted with swim-up stage salmon larvae in a standardized fresh water that simulated
dilution of San Luis Drain water.
Survival and growth (length and weight) were measured at 30, 60, and 90 days.
Unexplained control mortality (33%) between day 60 and day 90 introduced an unacceptable
level of uncertainty into the overall health of the fish. The 1985 Aquatic Life Guidelines
(USEPA, 1985) and the Manual of Instructions for Preparing Aquatic Life Water Quality Criteria
Documents (Stephan, 1987) require that excessive control mortality be treated as an exclusionary
threshold in data quality assessments for regulatory purposes such as deriving water quality
criteria. Therefore the 90 day survival data from this study was not used quantitatively. At 60
days, larval control mortality was acceptable (1%), and 60 day larval survival was > 90% in all
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SLD and SeMet treatments (3.2 ppm - 18.2 ppm) except for the high Se treatment (35.4 ppm).
Whole body selenium concentrations were measured at 60 days, were 10.4 and 13.3 mg/kg dw,
respectively, for larvae fed the SeMet and SLD diets of 18.2 mg/kg dw (Hamilton et al. 1990).
Although survival was similar in response to the two diets, larval growth responses
differed between the SLD and SeMet diets. The salmon fed the SeMet mosquitofish diet had
significant reductions in both length and weight at 30, 60, and 90 days; but only at the 2 highest
concentrations (18.2 and 35.4 ppm). The average length and weight of the larvae fed the SLD
mosquitofish diet were significantly lower at all concentrations at 30, 60, and 90 days. The
greater effect on growth parameters fed the SLD mosquitofish meal diet could have been caused
by one or more of several factors: 1) additional forms of organic selenium (e.g., selenocysteine)
present in the SLD mosquitofish, 2) additional toxic elements (e.g., heavy metals) that were
accumulated by the SLD mosquitofish, and not present in the reference site mosquitofish, and 3)
differential metabolic processing of the organoselenium contained in the proteins of the SLD
mosquitofish and fed to the larval salmon, versus the larvae fed the diet containing the free
amino acid selenomethionine (Hamilton et.al. 1990).
EPA performed a regression on the 60-day weight and whole body concentrations, and
derived a whole body EC 10 value of 7.355 jig/g dw for the SeMet diet for reduced growth, and a
whole body EC 10 value of 11.14 jig/g dw for the SLD diet for reduced growth. These values are
the only two available EC 10 Species Mean Chronic Values (SMCVs) for non-reproductive
endpoints for the genus Oncorhynchus, and the Genus Mean Chronic Value (GMCV) is 9.052
Hg/g dw.
EPA recommends that states and tribes consider use of the whole-body criterion element
for juvenile (smolt) anadromous Pacific salmon species as the primary criterion element over the
other elements due to the unique life history of these species, specifically, the lack of exposure to
adult salmonids from selenium in freshwater prior to reproduction. The hierarchal structure of
the egg-ovary tissue over the other tissue criterion elements applies to all other species in the
family Salmonidae. The egg-ovary criterion element, as well as the other fish tissue criterion
elements and the water column criterion elements still apply, as applicable, to protect the
remainder of the aquatic community in these waters.
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6.4 AQUATIC-DEPENDENT WILDLIFE is BEYOND THE SCOPE OF THIS AQUATIC
CRITERIA DERIVATION
AWQC that are developed by EPA typically focus directly on aquatic life, not aquatic-
dependent wildlife such as birds. As presented by Campbell (2011), EPA recognizes that
selenium effects on aquatic-dependent wildlife are also of concern but considers them beyond
the scope of this national criterion update. In the interest of providing updated guidance to
protect against the known risks of selenium exposure to fish, EPA decided to focus its analyses
on updating the existing selenium criterion for freshwater aquatic life based on the latest
scientific evidence.
EPA plans, in the future, to consider the effects of selenium on aquatic-dependent
wildlife, potentially in the form of criteria expanded to address aquatic-dependent wildlife. When
translated to a water concentration, a criterion protective of aquatic-dependent wildlife may be
more stringent or possibly less stringent, than the values provided for aquatic life in this 2015
criteria document. This is because data indicate that for most ecosystems, selenium
concentrations are generally conserved or increase incrementally at each trophic level in a food
web (after a substantial increase from water to trophic level 1 (e.g. algae). Certain specific
ecosystems (e.g., estuarine and marine systems more commonly) with mollusk-based food-webs
may create a pathway for more selenium to bioaccumulate, particularly in molluscivorous
predators (certain fish and aquatic bird species) since the available data indicate that molluscs
generally have a higher trophic transfer factor than other invertebrate taxa., This level of
bioaccumulation is typically lower, and in contrast to other bioaccumulative chemicals such as
mercury which have much greater biomagnification.
As stated previously, the single largest step in tissue selenium accumulation in aquatic
environments occurs at the base of the food web where algae and other microorganisms
accumulate selenium from water (Orr et al. 2012; Stewart et al. 2010). Mollusks such as mussels
and clams accumulate selenium to a much greater extent than planktonic crustaceans and insects
due to higher ingestion rates of both particulate-bound (algae) and dissolved selenium from the
water column through filter feeding, and these organisms have a lower selenium elimination
rate.(Luoma and Rainbow 2005). Thus, aquatic-dependent wildlife criteria for species that are
primarily molluscivores may have concentrations of concern that were not protected by the 2015
selenium criterion elements found in this document. The criteria values for aquatic-dependent
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wildlife would be expected to depend on the aquatic systems, species, and food webs considered,
as well as spatial and temporal considerations related to selenium exposure and breeding and
nesting seasons.
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vxEPA
United States Office of Water EPA 822-P-15-001
Environmental Protection 4304T July 2015
Agency
External Peer Review Draft
Aquatic Life Ambient Water Quality
Criterion for
Selenium - Freshwater
2015
(Appendices A-M)
U.S. Environmental Protection Agency
Office of Water
Office of Science and Technology
Washington, B.C.
-------
LIST OF APPENDICES
APPENDIX A: Selenium Chemistry A-l
Inorganic Selenium A-2
Organoselenium A-4
Departure from Thermodynamic Equilibrium A-5
Physical Distribution of Species in Surface Water A-5
APPENDIX B: Conversions B-l
Conversion of Wet to Dry Tissue Weight B-2
Methodology B-2
Derivation of tissue conversion factors B-3
CF values calculated directly from whole-body and egg-ovary selenium
measurements B-6
Muscle to egg-ovary conversion factors B-18
Muscle to whole-body conversion factors B-36
Derivation of Trophic Transfer Function values B-51
Methodology B-51
TTF values from physiological coefficients B-53
TTF values from field data B-60
Food web models used to calculate composite TTFs to translate the egg-ovary FCV to
water-column values B-138
APPENDIX C: Summaries of Chronic Studies Considered For Criteria Derivation C-l
APPENDIX D: Summary Studies of Non-Reproductive Effects D-l
Studies of Non-Reproductive Effects D-2
Comparison of Fish Chronic Criterion Element to the Most Sensitive Non-Reproductive
SMCV D-12
APPENDIX E: Other Data E-l
Selenite E-2
Selenate E-2
Other Data - Endangered Species E-l3
Other Data- Chronic Studies with Fish Species E-21
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Other Data- Chronic Studies with Invertebrate Species E-52
Other Data - Field Study West Virginia Impoundments E-56
Other Data - Nutritional Deficiency/Sufficiency Studies Containing Measured Selenium in
the Diet and Whole Body Fish Tissue E-57
APPENDIXF: Toxi city of Selenium to Aquatic Plants F-l
Selenite F-2
Selenate F-2
APPENDIX G: Unused Data G-l
APPENDIX H: Calculation of EF Values H-l
APPENDIX I: Observed versus Predicted Egg-Ovary Concentrations I-1
APPENDIX J: Supplementary information on Selenium Bioaccumulation in Aquatic
Animals J-l
1.0 Effects of Growth Rate on the Accumulation of Selenium in Fish J-2
2.0 Analysis of the Relative Contribution of Aqueous and Dietary Uptake on the
Bioaccumulation of Selenium J-3
3.0 Kinetics of Accumulation and Depuration: Averaging Period J-4
3.1 Background J-4
3.2 Approach for Modeling Effects of Time-Variable Se Concentrations J-5
3.2.1 Model Results J-8
3.2.2 Summary of Scenario Results J-12
APPENDIX K: Site-Specific Criteria K-l
1.0 Translating the concentration of selenium in tissue to a concentration in water K-2
1.1 Relating the Concentration of Selenium in Tissue and Water using the mechanistic
modeling approach K-4
1.2 Steps for deriving a site-specific water concentration value from the egg-ovary FCV
K-9
1.2.1 Identify the appropriate target fish species K-9
1.2.2 Model the food-web of the targeted fish species K-13
1.2.3 Identify appropriate TTF values K-14
1.2.4 Determine the appropriate EF value K-l 8
1.2.5 Determine the appropriate CF value K-20
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1.2.6 Translate the selenium egg-ovary FCV into a site-specific water
concentration value using Equation 18 K-23
1.3 Managing uncertainty using the mechanistic modeling approach K-23
1.4 Example calculations K-24
1.4.1 Example 1 K-24
1.4.2 Example 2 K-25
1.4.3 Example 3 K-26
1.4.4 Example 4 K-27
1.5.5 Example 5 K-28
1.5.6 Example 6 K-29
1.5.7 Example 7a K-30
1.5.7 Example 7b K-31
2.0 Translating the concentration of selenium in tissue to a concentration in water using
Bioaccumulation Factors (BAF) K-31
2.1 Summary of the BAF approach K-31
2.2 Managing uncertainty using the BAF approach K-32
3.0 Comparison of Mechanistic Bioaccumulation Modeling and BAF approaches K-35
APPENDIX L: Analytical Methods for measuring Selenium L-l
General considerations when measuring concentrations of selenium L-2
Analytical methods recommended for measuring selenium in water L-3
American Public Health Standard Method 3114 B L-3
EPA Method 200.8 L-4
EPA Method 200.9 L-4
Analytical methods available for measuring selenium in fish tissue L-5
Strong acid digestion L-6
Dry-ashing digestion L-6
APPENDIX M: Abbreviations M-l
Reference and site abbreviations M-2
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APPENDIX A: SELENIUM CHEMISTRY
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Selenium in aquatic ecosystems exists in a broad range of oxidation states: (+ VI) in selenates
(HSeO4~, SeO42") and selenic acid (H2SeO4), (+ IV) in selenites (HSeO3~, SeO32") and selenous acid
(H2SeO3), 0 in elemental selenium, and (-II) in selenides (Se2~, HSe"), hydrogen selenide (H2Se), and
organic selenides (R2Se). Selenium also shows some tendency to form catenated species like organic
diselenides (RseSeR). Within the normal physiological pH range and the reduction potential range
permitted by water, only Se, SeO32", HSeO3", and SeO42" can exist at thermodynamic equilibrium (Milne
1998). While ionic reactions are expected to be rapid in water, oxidation-reduction reactions may be slow,
and the possibility exists for the formation of HSe" in living systems and some environments where
anoxic conditions arise. The parallel behavior of comparable species of sulfur and selenium in living
systems has often been observed, but it is important to recognize that their chemical characteristics are
different in many ways. For instance, selenate is comparable to chromate in oxidizing strength and far
stronger than sulfate [£°(SeO42'/H2SeO3) = 1.15 V; £°(Cr2O72VCr3+) = 1.33V; £°(SO42'/H2SO3) = 0.200V
(standard potentials in acid solution: Weast 1969)], whereas selenide is a much stronger reducing agent
than sulfide [£°(Se/H2Se) = -0.36 V; £°[S/H2S ]= 0.14V)].
INORGANIC SELENIUM
Selenate usually predominates in well-aerated surface waters, especially those with alkaline
conditions. In spite of its oxidizing strength, selenate (SeO42~) exhibits considerable kinetic stability in the
presence of reducing agents (Cotton and Wilkinson 1988). The radius of SeO42" is comparable to that of
SO42" (Frausto da Silva and Williams 1991), and uptake by cells is expected to take place via the same ion
channels or permeases for both anions. Competition between sulfate and selenate uptake has been
observed in many species: algae (Riedel and Sanders 1996), aquatic plants (Bailey et al. 1995),
crustaceans (Ogle and Knight 1996), fungi (Gharieb et al. 1995), HeLa cells (Yan and Frenkel 1994), and
wheat (Richter and Bergmann 1993). Reduced selenate bioconcentration with increasing sulfate
concentration has been demonstrated in Daphnia magna (Hansen et al. 1993). A significant inverse
relationship was shown to exist between acute selenate toxicity to aquatic organisms and ambient sulfate
concentrations (Brix et al. 200la). Competition with selenate has also been observed for phosphate in
green algae (Riedel and Sanders 1996), and with chromate and tungstate in anaerobic bacteria (Oremland
etal. 1989).
Selenous acid species (HSeO3~ and SeO32") can predominate in solution under the moderately
oxidizing conditions encountered in oxygenated waters. Between pH 3.5 and 9.0 biselenite ion is the
predominant ion in water, and at pH values below 7.0, selenites are rapidly reduced to elemental selenium
under mildly reducing conditions (Faust 1981), situations that are common in bottom sediments.
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Most selenite salts are less soluble than the corresponding selenates. The extremely low solubility
of ferric selenite Fe2(SeO3)3 (Ks= 2.0 ± 1.7 x 10'31), and of the basic ferric selenite Fe2(OH)4SeO3 (Ks =
10~61 7), is important to the environmental cycling of selenium. Selenites also form stable adsorption
complexes with ferric oxides, forming complexes of even lower solubility than the ferric selenites. Under
certain conditions, selenite (in contrast to selenate) seems to be completely adsorbed in high amounts by
ferric hydroxide and, to a lesser extent, by aluminum hydroxide (Faust 1981). Coprecipitation techniques
have been applied for preconcentration of selenium in natural waters, using iron (III) hydroxides, which
coprecipitates selectively the selenite, but not the selenate, species in river and sea waters (Yoshii et al.
1977). Alum and iron coagulation precipitation can be used in water treatment processes to remove
selenite (Clifford et al. 1986). The low levels of selenium in ocean waters have been attributed to the
adsorption of selenite by the oxides of metals, such as iron and manganese (National Academy of
Sciences 1976).
Relative to selenate, selenite is more reactive because of its polar character, resulting from the
asymmetric electron density of the ion, its basicity (attraction to bond with proton), and its nucleophilicity
(attraction to bond to a nucleus using the lone pair electrons of the ion). No evidence has yet been
presented to show that HSeO3" or SeO32" is taken up intact into the cell interior. Evidence indicates that
selenite is reduced rapidly, even before uptake in some cases, making it difficult to distinguish between
uptake and metabolic processes (Milne 1998). Freshwater phytoplankton process selenate and selenite by
different mechanisms, leading to different concentrations within the cell, and the concentrations attained
are affected by various chemical and biological factors in the environment (Riedel et al. 1991). These
authors suggested that selenate is transported into the cell by a biological process with low affinity,
whereas selenite appears to be largely physically adsorbed. Contradictory evidence suggesting that
selenite uptake is enzymatically mediated was found with marine phytoplankton (Baines and Fisher
2001). Experimental results supporting the hypothesis that separate accumulation mechanisms for
selenate and selenite are present in D. magna have been published (Maier et al. 1993). However, while
some organisms appear to absorb selenite nonspecifically, specific transport systems exist in other
species. Sulfate competition is insignificant in the aquatic plant Ruppia maritima (Bailey et al. 1995), and
specific uptake systems have been demonstrated in some soft line microorganisms (Heider and Boeck
1993). Selenite uptake in green algae, unlike selenate, is increased substantially at lower pH values, a
property that represents another difference between these two anions (Riedel and Sanders 1996). The
uptake of inorganic selenium species, selenate and selenite, by the green alga Chlamydomonas reinhardtii
(Dang) was examined as a function of pH over the range 5 to 9, and in media with varying concentrations
of major ions and nutrients using 75Se as a radiotracer. Little difference was noted in the uptake of
selenate as a function of pH, with the maximum uptake found at pH 8; however, selenite uptake increased
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substantially at the lower pH values. Differences in speciation are suggested to be the cause of these
differences. Selenate exists as the divalent ion SeO42" over the range of pH tested; whereas monovalent
biselenite ion HSeO3" is prevelant at these pH values. At the low end of the pH range, neutral selenous
acid may also play a role.
Elemental selenium is not measurably soluble in water. It has been reported that elemental
selenium is slowly metabolized by several bacteria (Bacon and Ingledew 1989), and the translocation of
elemental selenium into the soft tissue of the marine mollusk Macoma balthica has been reported (Luoma
et al. 1992). The bioavailability of elemental selenium toM balthica was assessed by feeding the
organisms 75Se-labeled sediments in which the elemental selenium was precipitated by microbial
dissimilatory reduction. A 22% absorption efficiency of particulate elemental selenium was observed. In
view of the insolubility of elemental selenium, uptake may be preceded by air oxidation, or in reducing
environments thiols may facilitate the solubilization (Amaratunga and Milne 1994). Elemental selenium
can be the dominant fraction in sediments (Zawislanski and McGrath 1998).
Selenium is reduced to hydrogen selenide, H2Se, or other selenides at relatively low redox
potentials. Hydrogen selenide by itself is not expected to exist in the aquatic environment since the
Se°/H2Se couple falls even below the ETVH2 couple. Aqueous solutions of H2Se are actually unstable in air
due to its decomposition into elemental selenium and water. Under moderately reducing conditions,
heavy metals are precipitated as the selenides, which have extremely low solubilities. The following are
log Ks values of some heavy metal selenides of environmental interest: -11.5 (Mn2+), -26.0 (Fe2+), -60.8
(Cu+), -48.1 (Cu2+), -29.4 (Zn2+), -35.2 (Cd2+), and -64.5 (Hg2+). The precipitation of selenium as heavy
metal selenides can be an important factor affecting the cycling of the element in soils and natural waters.
ORGANOSELENIUM
Organic selenides (conventionally treated as Se(-II) species) in variable concentrations, usually in
the form of free and combined selenomethionine and selenocysteine, are also present in natural surface
waters (Fisher and Reinfelder 1991). Dissolved organic selenides may be an important source of selenium
for phytoplankton cells, because they can account for -80% of the dissolved selenium in open ocean
surface waters, and for a significant fraction in many other environments as well (Cutter 1989; Cutter and
Cutter 1995). Dissolved organoselenium levels of 14.2%, 65% and 66% were measured in samples (one
meter depth) from Hyco Reservoir, NC; Robinson Impoundment, SC; and Catfish Lake, NC; respectively
(Cutter 1986). The Hyco Reservoir organoselenium was identified as being protein bound.
Organoselenium concentrations were found to range from 10.4% (58.7 ug/L) to 53.7% (1.02 ug/L) of the
total selenium present in Lake Creek and Benton Lake, MT surface waters (Zhang and Moore 1996).
Organoselenium quite often is measured as the difference between total dissolved selenium and the sum
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of selenite plus selenate, and is therefore not typically characterized. Much more work is needed in the
area of specific identification and characterization of the nature of the organic selenides present in aquatic
ecosystems. Organoselenium form(s) are much more bioavailable and probably play a very important role
in selenium ecotoxic effects (e.g. Besser et al., 1993; Rosetta and Knight 1995).
DEPARTURE FROM THERMODYNAMIC EQUILIBRIUM
In the highly dynamic natural waters, there is often a departure from thermodynamic equilibrium.
In the thermodymanic models, kinetic barriers to equilibrium and biological processes are not adequately
considered, and the speciation of selenium in oxidized natural waters is not accurately predicted. Selenate
is usually the predominate form in solution; however, selenite and organoselenium can both exist at
concentrations higher than predicted (Faust 1981; Luoma et al. 1997). Bioaccumulation by
microorganisms, bioproduction and release of organoselenium, and mineralization of particulate selenium
forms contribute to the disequilibrium.
PHYSICAL DISTRIBUTION OF SPECIES IN SURFACE WATER
The physical distribution of various selenium species in surface waters is regulated by:
• sorption to or incorporation in suspended particulate matter (SPM), and
• complexation with inorganic and/or organic colloidal material, such as (FeO OH)n and humic
substances (dissolved organic matter, DOM).
Both sorption to SPM and complexation with colloidal matter reduces the bioavailability of the
selenium species. The average fraction of selenium associated with the suspended particulate phase
(0.45um filtration) as determined from eleven different studies of various surface waters was found to be
16% (0-39% range) of the total selenium, i.e., an average operationally defined dissolved selenium level
of 84% (Table A-l). In the James River, VA, the dissolved inorganic and organic selenium was found to
be 77% and 70% associated with colloidal matter, respectively (Takayangi and Wong 1984). A study of
lake ecosystems in Finland (Wang et al. 1995) found that 52% of the dissolved selenium was associated
with humic substances, and in a similar speciation study of Finnish stream waters, Lahermo et al. (1998)
determined that 36% of the selenium was complexed with humic matter. Hence, in various waterbodies
physical distribution as well as chemical speciation of selenium must be considered in relationship to
bioavailability and aquatic toxicity.
Until recently, the organic selenium fraction has been routinely measured as the difference
between total dissolved selenium and the sum of selenite and selenate. Unfortunately, the calculation of
this important selenium fraction in water as the difference between the total and measurable inorganic
fractions has not permitted this fraction to be fully characterized. New techniques are currently being
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developed which should help the specific identification and characterization of the nature of the organic
selenides present in aquatic systems. This work is particularly important because portions of the organic
selenium fraction (e.g., selenomethionine) of total dissolved selenium in water have been shown to be
much more bioavailable than the other forms of selenium, and therefore this work is also important for
understanding the manifestation of selenium ecotoxic effects.
Table A 1. Suspended particulate and dissolved selenium as a function of total selenium in
freshwater and marine aquatic ecoystems.
Reference
Cutter 1989
Cutter 1986
Tanizaki et al. 1992
Luomaetal. 1992
Cumbie and VanHorn, 1978
GLEC 1997
Wangetal. 1995
Lahermoetal. 1998
Hamilton etal. 200 la,b
Hamilton etal. 200 la,b
Hamilton et al. 2001a,b
Nakamoto and Hassler 1992
Nakamoto and Hassler 1992
Welsh and Maughan 1994
Welsh and Maughan 1994
Welsh and Maughan 1994
Welsh and Maughan 1994
Welsh and Maughan 1994
Welsh and Maughan 1994
Waterbody
Carquinezitist, CA
Hyco Reservoir, NC
Japanese Rivers
San Francisco Bay, CA
Belews Lake, NC
Unnamed Stream, Albright, WV
Finnish Lakes
Finnish Streams
Adobe Creek, Fruita, CO
North Pond, Fruita, CO
Fish Ponds, Fruita, CO
Merced River, CA
Salt Slough, CA
Cibola Lake, CA
Hart Mine Marsh, Blythe, CA
Colorado River, Blythe, CA
Palo Verda Oxbow Lake, CA
Palo Verda Oufall Drain, CA
Pretty Water Lake, CA
Particulate Se
(% of Total)
20-40
0
16
22-31
8
4
10
8
18
0
7
0
4
39
6
11
33
0
21
Fraction
dissolved, fd
0.6-0.8
1
0.84
0.69-0.78
0.92
0.96
0.9
0.92
0.82
1
0.93
1
0.96
0.62
0.94
0.89
0.67
1
0.79
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A-6
Draft Document
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APPENDIX B: CONVERSIONS
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CONVERSION OF WET TO DRY TISSUE WEIGHT
Methodology
Conversion factors (CF) derived from selenium measurements were calculated using
concentrations expressed as dry weights ((ig/g dry weight). The majority of tissue and whole-body
selenium concentrations were reported as dry weights. Measurements reported as wet weight were
converted to equivalent dry weights using available percent moisture data for the relevant species and
tissue type.
Species-specific percent moisture data for muscle tissue were available for bluegill (Gillespie and
Baumann 1986; Nakamoto and Hassler 1992), rainbow trout (Seiler and Skorupa 2001), and for a
composite average of nine fish species (May et al. 2000). Species specific percent moisture data for
ovaries were available for bluegill (Gillespie and Baumann 1986; Nakamoto and Hassler 1992), fathead
minnow (GEI Associates 2008; Rickwood et al. 2008), and rainbow trout (Seiler and Skorupa 2001).
Species-specific % moisture data for whole-body tissues were available for bluegill (USGS NCBP).
Measurements reported as wet weight were converted to equivalent dry weights using available
percent moisture data for the relevant species and tissue type. If percent moisture data were unavailable
for a fish species, percent moisture data for a similar species (i.e., same genus or, if unavailable, same
family) were used. Table B-l lists percent moisture by tissue type, species, data source, and the target
species and study for which the % moisture data were used to convert from wet to dry weight.
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Table B-l. Percent moisture, by species and tissue type.
% Moisture Data Source
Species
Study
% Moisture by Tissue
Whole-
body
Muscle
Ovary
Conversion Applied to
Species
Study
Used in derivation of FCV
Rainbow trout
Rainbow trout
Fathead
minnow
Bluegill
Avg of 9 spp
Seiler & Skorupa 2001
Seller &Skorupa 2001
Average of GEI Assoc. 2008;
Rickwood et al. 2008
Average of Gillespie &
Baumann 1986 and Nakamoto
&Hassler 1992
May et al. 2000
78.4
61.20
61.20
75.30
76.00
Rainbow trout
Brook trout
Fathead minnow
Bluegill
Striped bass
Holm et al. 2005
Holm et al. 2005
Schultz and
Hermanutz 1990
Hermanutz et al.
1996
Coughlan and Velte
1989
Used in conversion of FCV in egg/ovary to whole-body Se concentrations
Bluegill
Bluegill
Bluegill
Rainbow trout
Rainbow trout
Rainbow trout
Rainbow trout
Rainbow trout
Rainbow trout
USGS NCBP
May et al. 2000
Average of Gillespie &
Baumann 1986 and Nakamoto
&Hassler 1992
May et al. 2000
Seiler & Skorupa 2001
May et al. 2000
Seiler & Skorupa 2001
May et al. 2000
Seiler & Skorupa 2001
74.80
80.09
77.54
77.54
77.54
76.00
61.20
61.20
61.20
Bluegill
Bluegill
Bluegill
Brook Trout
Brook Trout
Rainbow Trout
Rainbow Trout
Rainbow Trout
Rainbow Trout
Hermanutz et al.
1996
Hermanutz et al.
1996
Hermanutz et al.
1996
Holm et al. 2005
Holm et al. 2005
Holm et al. 2005
Holm et al. 2005
Casey & Siwik 2000
Casey & Siwik 2000
DERIVATION OF TISSUE CONVERSION FACTORS
Methodology
EPA used a mechanistic bioaccumulation modeling approach to derive a mathematical
relationship between the concentration of selenium in water to the concentration of selenium in the eggs
and ovaries offish. This approach characterizes selenium bioaccumulation as a series of steps
representing the phase transformation of selenium from dissolved to particulate form, and then the trophic
transfer of selenium through aquatic food webs to invertebrates and fish. The final step in this process is
the transfer of selenium into eggs and ovary tissue.
Do not distribute, quote or cite
B-3
Draft Document
-------
Equation 1 quantitatively models the transfer of selenium through each environmental
compartment as a series of site-specific and species-specific parameters. The parameter CFin Equation 1
represents the species-specific proportion of selenium in egg or ovary tissue relative to the average
concentration of selenium in all body tissues and is given as:
C
-
egg-ovary
(Equation 1)
whole-body
where
CF = Whole-body to egg-ovary conversion factor (dimensionless ratio).
C egg-ovary = Selenium concentration in the eggs or ovaries of fish (|ig/g dw)
C Whoie-body = Selenium concentration in the whole body of fish (|ig/g dw).
EPA derived species-specific conversion factor (CF) values using the same methods that were
used to derive species-specific TTF values from field data. To derive whole-body to egg -ovary CF
values, the EPA defined matched pairs of selenium measurements from the whole-body and from the eggs
or ovaries measured from the same individual fish or from matched composite samples. Egg-ovary
concentration was defined as a measurement from either the eggs or the ovaries. If multiple measurements
from both eggs and ovaries of the same individual or matched composite sample were available, the
average value was used. EPA first confirmed a statistical relationship between egg-ovary and whole
body selenium for each species using ordinary least squares (OLS) linear regression. If the regression
resulted in a statistically significant (P<0.05) positive slope, EPA calculated the ratio of the egg -ovary to
whole body selenium concentration for each matched pair of measurements and used the median as the
CF value for that species.
EPA derived CF values from selenium measurements in units of (ig/g dry weight. The majority
of tissue and whole body selenium concentrations were reported as dry weights. Measurements reported
as wet weight were converted to equivalent dry weights using available percent moisture data for the
relevant species and tissue type. If percent moisture data were unavailable for a fish species, percent
moisture data for a similar species (i.e., same genus or, if unavailable, same family) were used. A listing
of percent moisture concentrations by species and target tissue are provided in the bottom portion of
Table B-l.
For those species without sufficient data to directly calculate an egg-ovary to whole body CF, but
which had sufficient data to calculate a conversion factor for either egg-ovary to muscle or whole body to
muscle, the EPA followed a two stage approach based on taxonomic similarity similar to that described
above. If a fish species had species specific egg -ovary to muscle conversion factor, but no whole body
Do not distribute, quote or cite B-4 Draft Document
-------
data with which to calculate an egg to whole body CF, then available data would be used to estimate a
muscle to whole body conversion factor for that species based on taxonomic relatedness. The estimated
muscle to whole body factor would be multipled by the directly measured egg-ovary to muscle factor to
estimate an egg-ovary to whole body CF for that species. For example, rainbow trout has a species
specific egg-ovary to muscle conversion factor of 1.92, but does not have a species specific egg-ovary to
whole body CF. Using the taxonomic approach described above, the most closely related taxa to rainbow
trout with muscle to whole body conversion factors are in the class Actinopterygii. The median
conversion factor for the 8 species within that class is 1.27. The final egg-ovary to whole body CF for
rainbow trout is 2.44 (Table 11), or 1.92 x 1.27.
The EPA developed species-specific egg-ovary to muscle and muscle to whole-body correction
factors following the procedure described for whole-body to egg-ovary conversion factors. The EPA
obtained matched pairs of selenium measurements in the whole-body and muscle filets and matched pairs
of selenium measurements in muscle filets and whole-body from published scientific literature. EPA first
confirmed a statistical relationship between the two tissue types for each species using OLS linear
regression. If the regression resulted in a significant fit with a positive slope, the EPA calculated the ratio
of each matched pair of measurements and then calculated the median ratio.
Do not distribute, quote or cite B-5 Draft Document
-------
CF values calculated directly from whole-body and egg-ovary selenium measurements
c,
^ whole-bocty
egg
^ ovary
*-- egg-ovary
= Selenium concentration in all tissues ((ig/g dw)
= Selenium concentration in eggs ((ig/g dw)
= Selenium concentration in ovary tissue ((ig/g dw)
= Average selenium concentration in eggs and ovaries
'c+c
egg-ovary
Kauo -
whole-body
Black bullhead (Ameiurus melas)
Study
Osmundson
Osmundson
Osmundson
Osmundson
Osmundson
Osmundson
Osmundson
Osmundson
Osmundson
Osmundson
Osmundson
Cp
whole-body ^
etal.
etal.
etal.
etal.
etal.
et
al.
etal.
etal.
etal.
etal.
etal.
60 -I
40 •
^-•egg-ovary
20 •
o •
c
A
°\
)
2007
2007
2007
2007
2007
2007
2007
2007
2007
2007
2007
^
oV
0
20 40
*-• whole-body
5
4
5
4
9
7
7
6
8
2
5
60
30
80
50
90
60
60
30
60
60
00
30
Not
egg *^ ovary
64.
35.
52.
56.
42.
38.
37.
34.
26.
56.
64.
Median ratio:
R2
F:
df:
P:
Cegg-ovary Ratio
30
40
80
00
80
70
30
30
40
70
30
6.29
: 0.37
64
35
52
56
42
38
37
34
26
56
64
30
40
80
00
80
70
30
30
40
70
30
12
7
9
11
4
5
5
5
3
28
12
13
38
60
43
46
09
11
20
07
35
13
4.67
8
0.046
used because negative
slope.
Do not distribute, quote or cite
B-6
Draft Document
-------
Bluegill (Lepomis macrochirus)
Study
Coyleetal. 1993
Coyleetal. 1993
Coyleetal. 1993
Coyleetal. 1993
Coyleetal. 1993
Doroshov et al. 1992
Doroshov et al. 1992
Doroshov et al. 1992
Doroshov et al. 1992
Hermanutz et al. 1996
Hermanutz et al. 1996
Hermanutz et al. 1996
Hermanutz et al. 1996
Hermanutz et al. 1996
Hermanutz et al. 1996
Hermanutz et al. 1996
Hermanutz et al. 1996
Hermanutz et al. 1996
Hermanutz et al. 1996
Hermanutz et al. 1996
Hermanutz et al. 1996
Hermanutz et al. 1996
Hermanutz et al. 1996
Hermanutz et al. 1996
Hermanutz et al. 1996
Hermanutz et al. 1996
Osmundson et
60 i
40 •
*- egg-ovary
20 •
al. 2007
o
y
0 a
/
o/
°Y
f/O O O
Q® °
«
0 20 40
/-<
*-• whole-body
Cp
whole-body ^e
0.90
2.90
4.90
7.20
16.00
1.60
5.50
9.30
19.30
1.50
18.10
1.90
2.80
12.30
9.40
1.50
4.90
21.00
24.30
5.00
9.50
6.60
1.80
4.20
10.30
13.80
8.80
60
<
1.90
7.30
13.00
22.80
41.30
2.80
8.30
19.50
38.40
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
Median
^ ovary
2.10
8.30
12.50
25.00
41.00
-
-
-
-
0.30
16.70
4.40
8.40
29.00
24.50
3.20
10.30
42.10
55.00
7.00
26.00
14.90
4.40
7.90
16.30
15.90
9.70
ratio:
R2:
F:
df:
P:
^egg-ovary
2.00
7.80
12.75
23.90
41.15
2.80
8.30
19.50
38.40
0.30
16.70
4.40
8.40
29.00
24.50
3.20
10.30
42.10
55.00
7.00
26.00
14.90
4.40
7.90
16.30
15.90
9.70
2.13
0.82
110.9
25
< 0.001
Ratio
2.22
2.69
2.60
3.32
2.57
1.75
1.51
2.10
1.99
0.20
0.92
2.32
3.00
2.36
2.61
2.13
2.10
2.00
2.26
1.40
2.74
2.26
2.44
1.88
1.58
1.15
1.10
Do not distribute, quote or cite
B-7
Draft Document
-------
Bluehead sucker (Catostomus discobolus)
Study
Osmundson et al. 2007
Osmundson et al. 2007
Osmundson et al. 2007
Osmundson et al. 2007
Osmundson et al. 2007
Osmundson et al. 2007
Osmundson et al. 2007
10 -I
r 5 •
^ egg-ova iy
/3
o
0 5
*-• whole -body
Brown trout
Study
(Salmo trutta)
Formation 201 1 Saratoga fish hatchery
Formation 201 1 Saratoga fish hatchery
Formation 201 1 Saratoga fish hatchery
Formation 201 1 Saratoga fish hatchery
Formation 201 1 Saratoga fish hatchery
Formation 201 1 Saratoga fish hatchery
Formation 201 1 Saratoga fish hatchery
Formation 201 1 Saratoga fish hatchery
Formation 201 1 Saratoga fish hatchery
Formation 2011
Formation 2011
Formation 2011
Formation 2011
Formation 2011
Formation 2011
Formation 2011
Formation 2011
Formation 2011
Cp
whole-bodv ^e
1.30
2.00
2.10
2.20
2.40
3.90
5.60
10
Cp
whole-bodv ^e
3.60
4.10
3.70
4.30
3.00
3.10
2.70
2.50
8.90
13.80
17.90
13.60
17.20
6.70
9.60
22.60
7.20
9.20
zg *^ovarv
2.40
4.20
3.70
4.00
4.10
7.10
8.10
Median ratio:
R2:
F:
df:
P:
zg *^ovarv
0.80
0.90
0.80
0.90
1.20
1.20
1.00
1.00
12.80
40.30
36.00
26.80
26.90
18.60
17.70
38.80
13.20
13.40
*^ egg-ovary
2.40
4.20
3.70
4.00
4.10
7.10
8.10
1.82
0.95
88.9
5
<0.001
*^ egg-ovary
0.80
0.90
0.80
0.90
1.20
1.20
1.00
1.00
12.80
40.30
36.00
26.80
26.90
18.60
17.70
38.80
13.20
13.40
Ratio
1.85
2.10
1.76
1.82
1.71
1.82
1.45
Ratio
0.22
0.22
0.22
0.21
0.40
0.39
0.37
0.40
1.44
2.92
2.01
1.97
1.56
2.78
1.84
1.72
1.83
1.46
Do not distribute, quote or cite
B-8
Draft Document
-------
Brown trout (Salmo trutta)
Formation 2011
Formation 2011
Formation 2011
Formation 2011
Formation 2011
Formation 2011
Formation 2011
Formation 2011
Formation 2011
Formation 2011
Formation 2011
Formation 2011
Formation 2011
Formation 2011
Formation 2011
Formation 2011
Osmundson et al. 2007
Osmundson et al. 2007
Osmundson et al. 2007
Osmundson et al. 2007
13.20
8.60
11.30
20.00
8.40
5.60
6.70
5.90
6.00
7.00
5.60
4.70
7.20
9.20
5.50
8.50
4.60
4.30
5.00
5.50
20.50
12.50
11.20
28.10
12.80
8.40
8.50
8.40
9.10
7.50
6.60
6.90
6.20
14.00
6.90
9.50
1.20
37.80
35.60
32.50
20.50
12.50
11.20
28.10
12.80
8.40
8.50
8.40
9.10
7.50
6.60
6.90
6.20
14.00
6.90
9.50
1.20
37.80
35.60
32.50
1.55
1.45
0.99
1.41
1.52
1.50
1.27
1.42
1.52
1.07
1.18
1.47
0.86
1.52
1.25
1.12
0.26
8.79
7.12
5.91
50
C 25 1
^egg-ova ly
25
*-• whole -body
50
Median ratio: 1.45
R2:
F:
df:
P:
0.47
31.3
36
<0.001
Do not distribute, quote or cite
B-9
Draft Document
-------
Channel catfish (Ictalurus punctatus)
Study
Osmundson
Osmundson
Osmundson
Osmundson
et
et
et
et
al.
al.
al.
al.
2007
2007
2007
2007
CP
whole-body ^egj
3.40
3.30
2.60
4.00
l ^ovarv
29
21
13
30
Cegg-ovarv Ratio
.50
.10
.70
.30
29.50
21.10
13.70
30.30
8
6
5
7
.68
.39
.27
.58
- egg-ovary
30 -
20 -
10 -
Median ratio: 6.98
R2: 0.82
F: 9.1
df: 2
P: 0.099
Not used because P > 0.05.
10 20
^-•whole-body
30
Common carp (Cyprinus carpio)
Study
Osmundson
Osmundson
Osmundson
Osmundson
Osmundson
C/~<
whole-body ^eg!
et
et
et
et
et
al.
al.
al.
al.
al.
2007
2007
2007
2007
2007
6
4
11
23
4
.30
.80
.70
.10
.10
g ^ovarv
12.
9.
16.
27.
9.
^ess-ovarv
10
40
30
30
90
12
9
16
27
9
.10
.40
.30
.30
.90
Ratio
1.
1.
1.
1.
2.
92
96
39
18
41
30 n
20 -
*-egg-o
gg-ovary
10 -
Median ratio: 1.92
10
20
30
-•whole-body
R2:
F:
df:
P:
0.96
584.8
3
<0.001
Do not distribute, quote or cite
B-10
Draft Document
-------
Cutthroat trout (Oncorhynchus clarkii)
Study
Hardy 2005
Hardy 2005
Hardy 2005
Hardy 2005
Hardy 2005
Hardy 2005
Hardy 2005
Hardy 2005
Hardy 2005
Hardy 2005
Formation 2012
Formation 2012
Formation 2012
Formation 2012
Formation 2012
Formation 2012
Formation 2012
Formation 2012
Formation 2012
Formation 2012
Formation 2012
Formation 2012
Formation 2012
Formation 2012
Formation 2012
Formation 2012 Henry Lake fish hatchery
Formation 2012 Henry Lake fish hatchery
Formation 2012 Henry Lake fish hatchery
Formation 2012 Henry Lake fish hatchery
Formation 2012 Henry Lake fish hatchery
Formation 2012 Henry Lake fish hatchery
Formation 2012 Henry Lake fish hatchery
Formation 2012 Henry Lake fish hatchery
Formation 2012 Henry Lake fish hatchery
Formation 2012 Henry Lake fish hatchery
Formation 2012 Henry Lake fish hatchery
Formation 2012 Henry Lake fish hatchery
Formation 2012 Henry Lake fish hatchery
Formation 2012 Henry Lake fish hatchery
Formation 2012 Henry Lake fish hatchery
Formation 2012 Henry Lake fish hatchery
Cp
whole-body ^es
0.70
2.60
2.80
6.40
1.20
4.60
5.90
9.10
11.40
5.60
2.56
16.3
20.7
19.4
17
16.7
25.7
8.17
9.07
8.63
16.6
19.4
21
18.6
22.5
0.4
0.45
0.44
0.36
0.5
0.36
0.44
0.28
0.44
0.43
0.31
0.23
0.72
0.73
0.91
0.85
g *^ ovary
1.00
3.80
5.50
18.00
1.60
7.80
6.60
5.10
5.20
16.00
3.43
17.6
27.9
29.7
22.3
14.6
47.6
22
15.4
11.4
12.7
40.1
30
35.6
30.5
1.65
2.03
2.48
1.36
2.33
0.83
2.26
.87
.98
.34
3.23
.58
.93
.79
2.06
1.74
^egg-ovary
1.00
3.80
5.50
18.00
1.60
7.80
6.60
5.10
5.20
16.00
3.43
17.6
27.9
29.7
22.3
14.6
47.6
22
15.4
11.4
12.7
40.1
30
35.6
30.5
1.65
2.03
2.48
1.36
2.33
0.83
2.26
.87
.98
.34
3.23
.58
.93
.79
2.06
1.74
Ratio
1.43
1.46
1.96
2.81
1.33
1.70
1.12
0.56
0.46
2.86
1.34
1.08
1.35
1.53
1.31
0.87
1.85
2.69
1.70
1.32
0.77
2.07
1.43
1.91
1.36
4.13
4.51
5.64
3.78
4.66
2.31
5.14
6.68
4.50
3.12
10.42
6.87
2.68
2.45
2.26
2.05
Do not distribute, quote or cite
B-ll
Draft Document
-------
Cutthroat trout (Oncorhynchus clarkii)
50 -
45 -
40 -
35 -
30 -
^-egg-ovary .-,<. _
20 -
15 -
r
0,
O
O
o / Median ratio:
Qff'O
/^* R2:
o /" o p.
o° o/" °0 df:
,X^°
0 10 20 30
^whole-body
1.96
0.83
194.3
39
<0.001
Flannelmouth sucker (Catostomus latipinnis)
Study
Osmundson et al. 2007
Osmundson et al. 2007
Osmundson et al. 2007
Osmundson et al. 2007
Osmundson et al. 2007
Osmundson et al. 2007
Osmundson et al. 2007
Cp
whole-body ^eg!
3.00
2.60
3.10
3.10
3.50
4.40
4.50
g ^ovarv
4.00
4.10
5.90
4.30
5.70
6.20
6.20
^ess-ovary
4.00
4.10
5.90
4.30
5.70
6.20
6.20
Ratio
1.33
1.58
1.90
1.39
1.63
1.41
1.38
r 4
egg-ovary
Median ratio: 1.41
R2:
F:
df:
P:
0.65
9.2
5
0.021
4
-1
-'whole-body
Do not distribute, quote or cite
B-12
Draft Document
-------
Green sunfish (Lepomis cyanellus)
Study
Osmundson et al. 2007
Osmundson et al. 2007
Osmundson et al. 2007
Osmundson et al. 2007
Osmundson et al. 2007
Osmundson et al. 2007
Osmundson et al. 2007
Osmundson et al. 2007
Osmundson et al. 2007
Osmundson et al. 2007
Osmundson et al. 2007
Osmundson et al. 2007
Osmundson et al. 2007
Osmundson et al. 2007
Osmundson et al. 2007
Osmundson et al. 2007
Osmundson et al. 2007
Osmundson et al. 2007
Osmundson et al. 2007
Osmundson et al. 2007
Osmundson et al. 2007
Osmundson et al. 2007
Osmundson et al. 2007
Osmundson et al. 2007
Osmundson et al. 2007
Osmundson et al. 2007
Osmundson et al. 2007
Osmundson et al. 2007
Osmundson et al. 2007
Osmundson et al. 2007
Osmundson et al. 2007
Osmundson et al. 2007
Osmundson et al. 2007
Osmundson et al. 2007
Osmundson et al. 2007
Osmundson et al. 2007
Osmundson et al. 2007
Osmundson et al. 2007
Cp
whole-bodv *-^egg
22.80
8.80
15.40
4.80
5.70
4.40
3.80
11.90
6.40
9.50
9.10
6.20
7.00
7.70
6.20
10.20
9.70
9.90
7.20
9.00
9.70
8.90
9.80
9.90
10.30
5.30
10.10
11.80
3.30
4.00
4.30
3.70
6.20
3.50
4.40
5.60
4.90
4.40
^ovarv
27.40
10.20
21.80
7.00
8.90
6.40
6.40
18.10
12.30
13.80
15.20
10.80
11.70
12.60
10.00
13.90
15.20
14.70
8.80
12.90
13.10
11.50
13.20
11.60
7.50
8.10
13.20
14.00
5.20
5.80
4.10
4.90
9.50
4.80
5.60
10.10
7.50
5.90
^egg-ovary
27.40
10.20
21.80
7.00
8.90
6.40
6.40
18.10
12.30
13.80
15.20
10.80
11.70
12.60
10.00
13.90
15.20
14.70
8.80
12.90
13.10
11.50
13.20
11.60
7.50
8.10
13.20
14.00
5.20
5.80
4.10
4.90
9.50
4.80
5.60
10.10
7.50
5.90
Ratio
1.20
1.16
1.42
1.46
1.56
1.45
1.68
1.52
1.92
1.45
1.67
1.74
1.67
1.64
1.61
1.36
1.57
1.48
1.22
1.43
1.35
1.29
1.35
1.17
0.73
1.53
1.31
1.19
1.58
1.45
0.95
1.32
1.53
1.37
1.27
1.80
1.53
1.34
Do not distribute, quote or cite
B-13
Draft Document
-------
Green sunfish (Lepomis cyanellus)
30
20 •
*-egg-ov;
:gg-ovary
10 -
Median ratio: 1.45
10
20
30
-•whole-body
R2:
F:
df:
P:
0.87
240.0
36
< 0.001
Roundtail chub (Gila robusta)
Study
Osmundson et al. 2007
Osmundson et al. 2007
Osmundson et al. 2007
Osmundson et al. 2007
Osmundson et al. 2007
Osmundson et al. 2007
Osmundson et al. 2007
Cp
whole-body ^es!
4.10
5.30
6.40
6.80
5.50
6.60
8.40
2 ^ovarv
7.90
10.80
15.20
14.10
10.60
18.00
17.80
^egs-ovarv
7.90
10.80
15.20
14.10
10.60
18.00
17.80
Ratio
1.93
2.04
2.38
2.07
1.93
2.73
2.12
20 n
10 -
'egg-ovary
10
*-• whole -body
Median ratio: 2.07
20
R2:
F:
df:
P:
0.80
20.4
5
0.004
Do not distribute, quote or cite
B-14
Draft Document
-------
Smallmouth bass (Micropterus dolomieu)
Study
Osmundson et al. 2007
Osmundson et al. 2007
Osmundson et al. 2007
Osmundson et al. 2007
Osmundson et al. 2007
Osmundson et al. 2007
12 i
c 6 •
*- egg-ovary
0 -
C
White sucker
Study
O jS
X
6
^-•whole-body
(Catostomus commersonii)
Osmundson et al. 2007
Osmundson et al. 2007
Osmundson et al. 2007
Osmundson et al. 2007
Osmundson et al. 2007
Osmundson et al. 2007
Osmundson et al. 2007
Osmundson et al. 2007
Osmundson et al. 2007
Osmundson et al. 2007
Osmundson et al. 2007
Osmundson et al. 2007
Osmundson et al. 2007
Osmundson et al. 2007
Osmundson et al. 2007
Osmundson et al. 2007
Osmundson et al. 2007
Osmundson et al. 2007
Osmundson et al. 2007
Osmundson et al. 2007
Cp
whole-body ^e
4.20
5.50
5.40
7.80
5.10
4.90
12
Cp
whole-body ^e
3.80
4.20
3.30
4.50
6.30
6.80
11.00
12.70
5.70
3.90
3.80
9.90
5.30
10.70
5.90
7.00
6.40
6.30
5.30
6.20
zg *^ ovary
6.00
8.00
6.50
11.00
7.10
8.80
Median ratio:
R2:
F:
df:
P:
zg *^ ovary
6.20
6.20
5.20
6.50
7.70
5.80
10.90
11.20
9.40
5.40
5.10
10.40
10.40
11.00
11.70
11.60
9.40
10.20
7.30
8.90
*^ egg-ovary
6.00
8.00
6.50
11.00
7.10
8.80
1.42
0.73
10.6
4
0.026
*^ egg-ovary
6.20
6.20
5.20
6.50
7.70
5.80
10.90
11.20
9.40
5.40
5.10
10.40
10.40
11.00
11.70
11.60
9.40
10.20
7.30
8.90
Ratio
1.43
1.45
1.20
1.41
1.39
1.80
Ratio
1.63
1.48
1.58
1.44
1.22
0.85
0.99
0.88
1.65
1.38
1.34
1.05
1.96
1.03
1.98
1.66
1.47
1.62
1.38
1.44
Do not distribute, quote or cite
B-15
Draft Document
-------
White sucker (Catostomus commersonii)
Osmundson et al. 2007
Osmundson et al. 2007
Osmundson et al. 2007
Osmundson et al. 2007
Osmundson et al. 2007
Osmundson et al. 2007
Osmundson et al. 2007
Osmundson et al. 2007
Osmundson et al. 2007
Osmundson et al. 2007
Osmundson et al. 2007
Osmundson et al. 2007
Osmundson et al. 2007
Osmundson et al. 2007
Osmundson et al. 2007
Osmundson et al. 2007
Osmundson et al. 2007
Osmundson et al. 2007
Osmundson et al. 2007
Osmundson et al. 2007
15 -
10 -
*-egg-ovary
5 -
°cS ^rf^l
CD o 3^ef^
°^XX**Cb
.ff^^ o o o
xg*S®
0
0.
0 5 10
/-<
^whole-body
5.60
8.80
8.70
11.40
10.70
8.40
7.00
7.50
10.30
6.70
2.10
1.80
3.20
2.30
3.10
3.00
2.80
2.50
3.40
2.80
•\
J
15
10.50
10.20
8.10
9.50
10.70
8.30
12.00
6.10
6.10
11.30
2.60
3.60
4.40
4.40
4.80
4.30
4.10
3.80
3.60
3.80
Median ratio:
R2:
F:
df:
P:
10.50
10.20
8.10
9.50
10.70
8.30
12.00
6.10
6.10
11.30
2.60
3.60
4.40
4.40
4.80
4.30
4.10
3.80
3.60
3.80
1.41
0.54
45.4
38
< 0.001
1.88
1.16
0.93
0.83
1.00
0.99
1.71
0.81
0.59
1.69
1.24
2.00
1.38
1.91
1.55
1.43
1.46
1.52
1.06
1.36
Do not distribute, quote or cite
B-16
Draft Document
-------
Table B-2. Summary of whole-body to egg-ovary conversion factors (CF) from matched
pairs of whole-body and egg-ovary measurements.
Common name
Bluegill
Bluehead sucker
Brown trout
Common carp
Flannelmouth sucker
Cutthroat trout
Green sunfish
Roundtail chub
Smallmouth bass
White sucker
Scientific name
Lepomis macrochirus
Catostomus discobolus
Salmo trutta
Cyprinus carpio
Catostomus latipinnis
Oncorhynchus clarkii
Lepomis cyanellus
Gila robusta
Micropterus dolomieu
Catostomus commersonii
Median ratio (CF)
2.13
1.82
1.45
1.92
1.41
1.96
1.45
2.07
1.42
1.41
Do not distribute, quote or cite
B-17
Draft Document
-------
Muscle to egg-ovary conversion factors
*-• muscle
ovary
c,
egg-ovary
Ratio
= Selenium concentration in muscle tissue only (|ig/g dw)
= Selenium concentration in eggs ((ig/g dw)
= Selenium concentration in ovary tissue ((ig/g dw)
= Average selenium concentration in eggs and ovaries
C.
egg-ovary
C
muscle
Hack bullhead (Ameiurus melas)
tudy
)smundson et al
)smundson et al
)smundson et al
)smundson et al
)smundson et al
)smundson et al
)smundson et al
)smundson et al
)smundson et al
)smundson et al
80 -
60 -»
r 40 •
egg-ovary
20 •
0 i
C/~<
muscle ^t
. 2007 3.40
. 2007 3.90
. 2007 4.30
. 2007 4.70
. 2007 5.70
. 2007 7.40
. 2007 7.50
. 2007 7.80
. 2007 7.80
. 2007 9.20
o o
~
o $£
O
~™"™™™™""™^
0 2 4 6 8 10
^ muscle
Not
gs ^ovarv
64.30
35.40
52.80
56.00
42.80
38.70
37.30
34.30
26.40
56.70
Median ratio:
R2:
F:
df:
P:
used because P >
slope.
^ess-ovarv
64.30
35.40
52.80
56.00
42.80
38.70
37.30
34.30
26.40
56.70
6.84
0.17
1.65
8
0.250
Ratio
18.91
9.08
12.28
11.91
7.51
5.23
4.97
4.40
3.38
6.16
0.05 and negative
Do not distribute, quote or cite
B-18
Draft Document
-------
Bluegill (Lepomis macrochirus)
Study
muscle
Brysonetal. 1984
Brysonetal. 1985a(pt. 1)
Brysonetal. 1985a(pt. 1)
Brysonetal. 1985a(pt. 2)
Doroshov et al.
Doroshov et al.
Doroshov et al.
Doroshov et al.
Hermanutz et al
Hermanutz et al
Hermanutz et al
Hermanutz et al
Hermanutz et al
Hermanutz et al
Hermanutz et al
Hermanutz et al
Hermanutz et al
Hermanutz et al
Hermanutz et al
Hermanutz et al
Hermanutz et al
Hermanutz et al
Hermanutz et al
Hermanutz et al
Hermanutz et al
Hermanutz et al
Hermanutz et al
Hermanutz et al
1992
1992
1992
1992
1996
1996
1996
1996
1996
1996
1996
1996
1996
1996
1996
1996
1996
1996
1996
1996
1996
1996
1996
1996
Osmundson et al. 2007
60 -I
40 -
c
'"egg-ovary
20 -
Q
o ^^
^^ o
0 °/^
o ^^ o
8°
cP— rfT
o><»
ft&O O
0 20 40 60 80 100
C ,
^muscle
err
*^egg ^^ ovary ^^ egg-ovary
84.0 - 49.0 49.0
59.0 - 30.0 30.0
2.7 - 2.2 2.2
25.0 - 9.1 9.1
1.5 2.8 - 2.8
5.8 8.3 - 8.3
10.4 19.5 - 19.5
23.6 38.4 - 38.4
1.6 - 2.0 2.0
8.5 - 18.8 18.8
14 - 15.5 15.5
2.1 - 0.3 0.3
20.6 - 16.7 16.7
1.9 - 4.4 4.4
3.5 - 8.4 8.4
17.6 - 29.0 29.0
12.5 - 24.5 24.5
2.3 - 3.2 3.2
6.9 - 10.3 10.3
44.9 - 42.1 42.1
39.8 - 55.0 55.0
5.3 - 7.0 7.0
12.5 - 26.0 26.0
7.8 - 14.9 14.9
3.2 - 4.4 4.4
6.1 - 7.9 7.9
18.7 - 16.3 16.3
15.1 - 15.9 15.9
12.9 - 9.7 9.7
Median ratio: 1.38
R2: 0.65
F: 50.37
df: 27
P: <0.001
Ratio
0.58
0.51
0.81
0.36
1.87
1.43
1.88
1.63
1.25
2.21
1.11
0.14
0.81
2.32
2.40
1.65
1.96
1.39
1.49
0.94
1.38
1.32
2.08
1.91
1.38
1.30
0.87
1.05
0.75
Do not distribute, quote or cite
B-19
Draft Document
-------
Bluehead sucker (Catostomus discobolus)
Study
muscle
Osmundson et al. 2007
Osmundson et al. 2007
Osmundson et al. 2007
Osmundson et al. 2007
Osmundson et al. 2007
Osmundson et al. 2007
Osmundson et al. 2007
10 -I
'"egg-ovary
0 -
c
^*^>
^^^
o
5 10
p
^muscle
err
*^egg ^^ ovary ^^ egg-ovary
1.5 - 2.4 2.4
2.3 - 4.2 4.2
2.5 - 3.7 3.7
2.7 - 4 4
3.1 - 4.1 4.1
5.2 - 7.1 7.1
8.6 - 8.1 8.1
Median ratio: 1.48
R2: 0.91
F: 47.70
df: 5
P: <0.001
Ratio
1.60
1.83
1.48
1.48
1.32
1.37
0.94
Brook trout (Salvelinus fontinalis)
Study
muscle
Holm et al. 2005
Holm et al. 2005
Holm et al. 2005
Holm et al. 2005
Holm et al. 2005
Holm et al. 2005
Holm et al. 2005
C C C
^^egg *" ovary *" egg-ovary
2.80 1.50 - 1.50
1.40 2.50 - 2.50
2.20 3.40 - 3.40
2.00 4.70 - 4.70
2.20 2.90 - 2.90
5.00 5.60 - 5.60
9.70 9.90 - 9.90
Holm et al. 2005 10.50 15.40 - 15.40
Holm et al. 2005 1
Holm et al. 2005 1
1.20 12.80 - 12.80
1.40 14.80 - 14.80
Holm et al. 2005 12.30 12.20 - 12.20
Holm et al. 2005 15.90 12.40 - 12.40
Holm et al. 2005 16.50 13.20 - 13.20
Holm et al. 2005 19.60 15.50 - 15.50
Holm et al. 2005 20.40 15.30 - 15.30
Holm et al. 2005 23.40 25.40 - 25.40
Holm et al. 2005 34.70 32.50 - 32.50
Ratio
0.54
1.79
1.55
2.35
1.32
1.12
1.02
1.47
1.14
1.30
0.99
0.78
0.80
0.79
0.75
1.09
0.94
Do not distribute, quote or cite
B-20
Draft Document
-------
Brook trout (Salvelinusfontinalis)
40 n
egg-ovary
20 -
Median ratio: 1.09
R2:
F:
df:
P:
0.91
152.3
15
< 0.001
0 20 40
*-• muscle
Brown trout (Salmo trutta)
otliCly ^muscle ^egg ^ovary ^egg-ovary
Osmundson et al. 2007 3.2 - 1.2 1.2
Osmundson et al. 2007 3.6 - 37.8 37.8
Osmundson et al. 2007 4 - 35.6 35.6
Osmundson et al. 2007
40 -,
30 -
C 20 -
'"egg-ovary
10 -
0 -
(
°° /
^<^ O
tt——., ,
) 5 10
r
^muscle
6.3 - 32.5 32.5
Median ratio: 7.03
R2: 0.17
F: 0.40
df: 2
P: 0.71
Not used because P > 0.05.
Ratio
0.38
10.50
8.90
5.16
Do not distribute, quote or cite
B-21
Draft Document
-------
Channel catfish (Ictaluris punctatus)
Study
"egg-ovary
30
20
10
0
o 9
C
muscle
P
^ov
Ratio
Osmundson et al. 2007
Osmundson et al. 2007
Osmundson et al. 2007
Osmundson et al. 2007
40
3.4
3.6
3.7
5.3
29.5
21.1
13.7
30.3
29.5
21.1
13.7
30.3
8.68
5.86
3.70
5.72
10
Median ratio: 5.79
R2: 0.20
F: 0.49
df: 2
P: 0.67
Not used because P > 0.05.
Common carp (Cyprinus carpio)
Study
Garcia-Hernandez 2000
Osmundson et al.
Osmundson et al.
Osmundson et al.
Osmundson et al.
Osmundson et al.
30 -
20 -
c
'"egg-ovary
10
2007
2007
2007
2007
2007
.X
o .X**^
o&^
o
0 10 20
*" muscle
CP P P
muscle ^egg ^ovarv ^egg-ovarv
4.6 - 1.8 1.8
7.8 - 12.1 12.1
8.2 - 9.4 9.4
20 - 16.3 16.3
24.2 - 27.3 27.3
6.6 - 9.9 9.9
^ Median ratio: 1.14
R2: 0.84
F: 21.7
df: 4
P: 0.007
30
Ratio
0.39
1.55
1.15
0.82
1.13
1.50
Do not distribute, quote or cite
B-22
Draft Document
-------
Cutthroat trout (Oncorhynchus clarkii)
Study
Colder 2005
Colder 2005
Colder 2005
Colder 2005
Colder 2005
Colder 2005
Colder 2005
Colder 2005
Colder 2005
Colder 2005
Colder 2005
Colder 2005
Colder 2005
Colder 2005
Colder 2005
Colder 2005
Colder 2005
Colder 2005
Colder 2005
Colder 2005
Colder 2005
Colder 2005
Colder 2005
Colder 2005
Colder 2005
Colder 2005
Colder 2005
Colder 2005
Colder 2005
Colder 2005
Colder 2005
Kennedy et al.
Kennedy et al.
Kennedy et al.
Kennedy et al.
Kennedy et al.
Kennedy et al.
Kennedy et al.
Kennedy et al.
Kennedy et al.
Kennedy et al.
Kennedy et al.
2000
2000
2000
2000
2000
2000
2000
2000
2000
2000
2000
CP
muscle *^e$
6.80
4.20
3.00
4.90
4.50
4.00
5.00
5.00
5.00
8.40
8.30
7.00
6.60
8.40
9.80
8.50
16.00
7.00
8.00
7.00
7.00
9.00
7.00
7.00
8.00
9.80
7.00
9.00
7.00
8.00
10.00
41.30
15.30
14.10
12.50
13.70
14.30
9.50
9.40
8.70
9.50
10.20
!g
75
58
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
.40
.40
30.60
20
19
16
16
14
.20
.40
.20
.10
.40
13.20
12
.60
12.30
^ ovary
28.
47.
22.
9.
8.
7.
10.
10.
8.
16.
18.
14.
14.
14.
16.
15.
20.
14.
19.
14.
14.
16.
13.
14.
14.
20.
22.
16.
12.
13.
14.
66.
31.
31.
18.
19.
16.
19.
22.
17.
13.
14.
20
80
00
80
20
00
00
00
00
20
30
30
30
70
40
90
00
00
00
00
00
00
00
00
00
20
00
00
00
00
00
80
60
40
50
50
20
30
00
00
60
50
^egg-ovary
28.20
47.80
22.00
9.80
8.20
7.00
10.00
10.00
8.00
16.20
18.30
14.30
14.30
14.70
16.40
15.90
20.00
14.00
19.00
14.00
14.00
16.00
13.00
14.00
14.00
20.20
22.00
16.00
12.00
13.00
14.00
71.10
45.00
31.00
19.35
19.45
16.20
17.70
18.20
15.10
13.10
13.40
Ratio
4.
11.
7.
2.
1.
1.
2.
2.
1.
1.
2.
2.
2.
1.
1.
1.
1.
2.
2.
2.
2.
1.
1.
2.
1.
2.
3.
1.
1.
1.
15
38
33
00
82
75
00
00
60
93
20
04
17
75
67
87
25
00
38
00
00
78
86
00
75
06
14
78
71
63
1.40
1.
2.
2.
1.
72
94
20
55
1.42
1.
1.
1.
1.
1.
1.
13
86
94
74
38
31
Do not distribute, quote or cite
B-23
Draft Document
-------
Cutthroat trout (Oncorhynchus clarkii)
Study
Kennedy et al.
Kennedy et al.
Kennedy et al.
Kennedy et al.
Kennedy et al.
2000
2000
2000
2000
2000
Rudolph et al. 2007
Rudolph et al. 2007
Rudolph et al. 2007
Rudolph et al. 2007
Rudolph et al. 2007
Rudolph et al. 2007
Rudolph et al. 2007
Rudolph et al. 2007
Rudolph et al. 2007
Rudolph et al. 2007
Rudolph et al. 2007
Rudolph et al. 2007
Rudolph et al. 2007
Rudolph et al. 2007
Rudolph et al. 2007
Rudolph et al. 2007
Rudolph et al. 2007
Rudolph et al. 2007
Rudolph et al. 2007
Rudolph et al. 2007
Rudolph et al. 2007
Rudolph et al. 2007
150 -I
100 -
^egg-ovary
50 •
n .
0
0
muscle
10.70
6.60
9.70
10.90
6.90
7.70
8.20
8.00
8.10
6.60
8.50
7.20
7.30
7.60
8.70
8.20
7.90
7.60
11.80
40.40
46.10
50.40
34.70
39.00
35.40
11.30
13.40
c c
^egg ^ovarv
10.50 20.60
9.90 21.50
9.10 13.20
8.50 13.40
13.20 20.30
13.90
12.50
15.00
14.90
15.20
12.90
12.30
16.70
13.10
15.60
13.90
15.10
12.30
16.10
86.30
121.00
140.00
51.00
65.30
46.80
16.90
20.60
Median ratio:
R2:
F:
df:
P:
^egg-ovary
15.55
15.70
11.15
10.95
16.75
13.90
12.50
15.00
14.90
15.20
12.90
12.30
16.70
13.10
15.60
13.90
15.10
12.30
16.10
86.30
121.00
140.00
51.00
65.30
46.80
16.90
20.60
1.81
0.82
308.3
67
< 0.001
Ratio
1.45
2.38
1.15
1.00
2.43
1.81
1.52
1.88
1.84
2.30
1.52
1.71
2.29
1.72
1.79
1.70
1.91
1.62
1.36
2.14
2.62
2.78
1.47
1.67
1.32
1.50
1.54
50
100
150
uscle
Do not distribute, quote or cite
B-24
Draft Document
-------
Dolly varden (Salvelinus malma)
Study
Colder 2009
Colder 2009
Colder 2009
Colder 2009
Colder 2009
Colder 2009
Colder 2009
Colder 2009
Colder 2009
Colder 2009
Colder 2009
Colder 2009
Colder 2009
Colder 2009
Colder 2009
Colder 2009
Colder 2009
200 -|
150 -
100 -
^egg-ovary
50 •
ft -
muscle
73.00
45.90
107.00
97.20
114.00
115.00
79.60
9.90
3.40
5.30
2.80
4.90
6.60
55.70
58.30
39.50
50.50
o
Cegg
^ovarv
92.30
40
107
102
124
185
112
7
12
70
00
00
00
00
00
00
10
9.60
5
10
11
65
51
60
56
40
50
00
80
90
50
60
Median ratio:
/>
R2:
F:
df:
P:
^egg-ovary
92.30
40
107
102
124
185
112
7
12
70
00
00
00
00
00
00
10
9.60
5
10
11
65
51
60
56
1.26
0.90
140.3
15
40
50
00
80
90
50
60
Ratio
1
0
1
1
1
1
26
89
00
05
09
61
1.41
0
3
1
1
2
1
1
0
1
1
71
56
81
93
14
67
18
89
53
12
< 0.001
0 50 100 150 200
muscle
Do not distribute, quote or cite
B-25
Draft Document
-------
^lannelmouth sucker (Catostomus latipinnis)
Study
3smundson
3smundson
3smundson
3smundson
3smundson
3smundson
3smundson
c
'"egg-ovary
muscle
et al. 2007
et al. 2007
et al. 2007
et al. 2007
et al. 2007
et al. 2007
et al. 2007
10 -I
5 -
0 -
o^>-o
irf (J
—————— -~~^^
0 5 10
^ muscle
err
*^egg *^ ovary *^ egg-ovary
3.6 - 4.0 4.0
3.8 - 4.1 4.1
4.1 - 5.9 5.9
4.6 - 4.3 4.3
5.2 - 5.7 5.7
6.2 - 6.2 6.2
7.3 - 6.2 6.2
Median ratio: 1.08
R2: 0.58
Ff C\^
: 6.92
df: 5
P: 0.036
Ratio
1.11
1.08
1.44
0.93
1.10
1.00
0.85
jreen sunfish (Lepomis cyanellus)
Study
3smundson
3smundson
3smundson
3smundson
3smundson
3smundson
3smundson
3smundson
3smundson
3smundson
3smundson
3smundson
3smundson
3smundson
3smundson
3smundson
3smundson
3smundson
3smundson
3smundson
muscle
et al. 2007
et al. 2007
et al. 2007
et al. 2007
et al. 2007
et al. 2007
et al. 2007
et al. 2007
et al. 2007
et al. 2007
et al. 2007
et al. 2007
et al. 2007
et al. 2007
et al. 2007
et al. 2007
et al. 2007
et al. 2007
et al. 2007
et al. 2007
C C C
'"egg '"ovary '"egg-ovary
28.1 - 27.4 27.4
12.9 - 10.2 10.2
21.9 - 21.8 21.8
5-77
6.1 - 8.9 8.9
5.2 - 6.4 6.4
5.1 - 6.4 6.4
15.7 - 18.1 18.1
10.1 - 12.3 12.3
11.5 - 13.8 13.8
10.5 - 15.2 15.2
7.2 - 10.8 10.8
9.3 - 11.7 11.7
7.7 - 12.6 12.6
6 - 10 10
12 - 13.9 13.9
12.1 - 15.2 15.2
12.5 - 14.7 14.7
7.5 - 8.8 8.8
11.3 - 12.9 12.9
Ratio
0.98
0.79
1.00
1.40
1.46
1.23
1.25
1.15
1.22
1.20
1.45
1.50
1.26
1.64
1.67
1.16
1.26
1.18
1.17
1.14
Do not distribute, quote or cite
B-26
Draft Document
-------
Jreen sunfish (Lepomis cyanellus)
>tudy
Dsmundson et al. 2007
Dsmundson et al. 2007
Dsmundson et al. 2007
Dsmundson et al. 2007
Dsmundson et al. 2007
Dsmundson et al. 2007
Dsmundson et al. 2007
Dsmundson et al. 2007
Dsmundson et al. 2007
Dsmundson et al. 2007
Dsmundson et al. 2007
Dsmundson et al. 2007
Dsmundson et al. 2007
Dsmundson et al. 2007
Dsmundson et al. 2007
Dsmundson et al. 2007
Dsmundson et al. 2007
Dsmundson et al. 2007
30 -I
20 -
r
'"egg-ovary
10 -
o .
J^
ojf^vf
<&r°
U T i r
0 10 20
'"muscle
Cp
muscle ^es
13.6
13.2
12.4
12.5
8.6
5.3
11.9
13.6
3.8
4.2
4.1
4.2
5.7
4.4
3.5
5.5
5
4.3
^
30
2 ^ovarv
13.1
11.5
13.2
11.6
7.5
8.1
13.2
14
5.2
5.8
4.1
4.9
9.5
4.8
5.6
10.1
7.5
5.9
Median ratio:
R2:
F:
df:
P:
^egg-ovary
13.1
11.5
13.2
11.6
7.5
8.1
13.2
14
5.2
5.8
4.1
4.9
9.5
4.8
5.6
10.1
7.5
5.9
1.21
0.89
281.4
36
<0.001
Ratio
0.96
0.87
1.06
0.93
0.87
1.53
1.11
1.03
1.37
1.38
1.00
1.17
1.67
1.09
1.60
1.84
1.50
1.37
Do not distribute, quote or cite
B-27
Draft Document
-------
Mountain whitefish (Prosopium williamsoni)
Study
Colder 2005
Colder 2005
Colder 2005
Colder 2005
Colder 2005
Colder 2005
Colder 2005
Colder 2005
Colder 2005
Colder 2005
Colder 2005
Colder 2005
Colder 2005
Colder 2005
Colder 2005
Colder 2005
Colder 2005
Colder 2005
Colder 2005
Colder 2005
Colder 2005
Colder 2005
Colder 2005
Colder 2005
Colder 2005
Colder 2005
Colder 2005
50 i
c 25 •
'"egg-ovary
0 -
(
muscle
3
3
3
4
3
3
5
5
5
7
7
5
7
3
4
4
5
4
10
4
7
6
6
5
6
5
4
jy0
f
) 25 50
^ muscle
60
70
10
20
90
50
20
00
20
60
20
50
80
70
70
40
70
00
00
90
60
10
80
00
60
00
80
C C
*"egg *"ovarv
26.
25.
20.
19.
19.
23.
38.
41.
32.
34.
32.
40.
39.
20.
22.
28.
30.
31.
35.
26.
26.
29.
41.
29.
34.
36.
28.
Median ratio:
R2
F:
df:
P:
90
80
00
30
20
20
00
00
00
00
00
00
70
30
40
90
10
50
20
70
80
70
10
00
50
30
90
'"egg-ova
26
25
20
19
19
23
38
41
32
34
32
40
39
20
22
28
30
31
35
26
26
29
41
29
34
36
28
5.80
0.33
12.4
25
<0.001
rv
90
80
00
30
20
20
00
00
00
00
00
00
70
30
40
90
10
50
20
70
80
70
10
00
50
30
90
Ratio
7
6
6
4
4
6
7
8
6
4
4
7
5
5
4
6
5
7
3
5
3
4
6
5
5
7
6
47
97
45
60
92
63
31
20
15
47
44
27
09
49
77
57
28
88
52
45
53
87
04
80
23
26
02
Do not distribute, quote or cite
B-28
Draft Document
-------
Northern pike (Esox lucius)
Study
Muscatello
Muscatello
Muscatello
Muscatello
Muscatello
Muscatello
Muscatello
Muscatello
Muscatello
Muscatello
Muscatello
Muscatello
Muscatello
Muscatello
etal.
etal.
etal.
etal.
etal.
etal.
etal.
etal.
etal.
etal.
etal.
etal.
etal.
etal.
60 -|
^egg-ovary
30-
0 -
C
*
2006
2006
2006
2006
2006
2006
2006
2006
2006
2006
2006
2006
2006
2006
o ^X^
X
/
X
30
^muscle
muscle
0
1
Ce
.90
.90
2.60
1
1
17
16
16
.30
.00
.00
.50
.50
2.00
2.00
1
.30
2.50
1
47
X
X
o
60
.30
.80
zg *^ ovary
3.50
2.70
3.40
3.70
2.70
43.20
24.50
26.10
3.40
4.10
4.10
4.10
3.40
48.20
Median ratio:
R2:
F:
df:
P:
^egg-ovary
3.50
2.70
3.40
3.70
2.70
43.20
24.50
26.10
3.40
4.10
4.10
4.10
3.40
48.20
1.88
0.83
58.9
12
<0.001
Ratio
3.89
1.42
1.31
2.85
2.70
2.54
1.48
1.58
1.70
2.05
3.15
1.64
2.62
1.01
Rainbow trout (Oncorhynchus mykiss)
Study
Casey and
Casey and
Casey and
Casey and
Casey and
Casey and
Casey and
Casey and
Casey and
Casey and
Casey and
Casey and
Siwik
Siwik
Siwik
Siwik
Siwik
Siwik
Siwik
Siwik
Siwik
Siwik
Siwik
Siwik
2000
2000
2000
2000
2000
2000
2000
2000
2000
2000
2000
2000
muscle
4
3
2
3
2
2
2
2
3
4
1
2
Ce
.10
.80
.60
.30
.30
.80
.30
.80
.00
.90
.50
.60
;g l^ ovary
11.60
10.10
0.10
4.90
3.60
5.30
3.70
6.40
5.20
6.80
3.60
6.90
^egg-ovary
11.60
10.10
0.10
4.90
3.60
5.30
3.70
6.40
5.20
6.80
3.60
6.90
Ratio
2
2
0
1
1
1
1
2
1
1
2
2
83
66
04
48
57
89
61
29
73
39
40
65
Do not distribute, quote or cite
B-29
Draft Document
-------
Rainbow trout (Oncorhynchus mykiss)
Study
Casey and Siwik 2000
Casey and Siwik 2000
Casey and Siwik 2000
Casey and Siwik 2000
Casey and Siwik 2000
Casey and Siwik 2000
Casey and Siwik 2000
Casey and Siwik 2000
Casey and Siwik 2000
Casey and Siwik 2000
Casey and Siwik 2000
Casey and Siwik 2000
Casey and Siwik 2000
Casey and Siwik 2000
Casey and Siwik 2000
Casey and Siwik 2000
Casey and Siwik 2000
Casey and Siwik 2000
Casey and Siwik 2000
Casey and Siwik 2000
Casey and Siwik 2000
Casey and Siwik 2000
Casey and Siwik 2000
Casey and Siwik 2000
Casey and Siwik 2000
Casey and Siwik 2000
Casey and Siwik 2000
Casey and Siwik 2000
Holm et al. 2005
Holm et al. 2005
Holm et al. 2005
Holm et al. 2005
Holm et al. 2005
Holm et al. 2005
Holm et al. 2005
Cp
muscle ^ej
4.60
4.60
3.60
2.40
3.70
2.70
0.70
0.60
0.60
28.60
30.90
32.40
28.00
31.70
29.50
30.10
29.90
32.80
31.40
32.00
35.70
24.60
30.30
25.70
35.00
33.80
28.70
25.80
1.70
1.60
1.30
4.00
4.30
8.50
7.40
zg *^ovarv
6.90
6.40
5.50
10.50
7.60
4.10
1.10
0.90
1.30
56.30
56.00
71.50
61.30
54.50
56.80
57.90
64.70
46.60
56.50
67.50
59.40
48.70
69.10
43.50
58.10
59.20
55.00
49.00
1.00
3.50
4.60
12.80
17.10
17.50
29.70
^egg-ovary
6.90
6.40
5.50
10.50
7.60
4.10
1.10
0.90
1.30
56.30
56.00
71.50
61.30
54.50
56.80
57.90
64.70
46.60
56.50
67.50
59.40
48.70
69.10
43.50
58.10
59.20
55.00
49.00
1.00
3.50
4.60
12.80
17.10
17.50
29.70
Ratio
1.50
1.39
1.53
4.38
2.05
1.52
1.57
1.50
2.17
1.97
1.81
2.21
2.19
1.72
1.93
1.92
2.16
1.42
1.80
2.11
1.66
1.98
2.28
1.69
1.66
1.75
1.92
1.90
0.59
2.19
3.54
3.20
3.98
2.06
4.01
Do not distribute, quote or cite
B-30
Draft Document
-------
Rainbow trout (Oncorhynchus mykiss)
Study
80 -,
p 40 -
'"egg-ovary
0
y
OJfl
0_
0
jre*
/
40
^" muscle
CP P
muscle *^ egg *^ ovary
Median ratio:
R2:
F:
df:
P:
80
^egg-ovary
1.92
0.96
990.0
45
<0. 001
Ratio
Razorback sucker (Xyrauchen texanus)
Study
Hamilton et al. 2005
Hamilton et al. 2005
Hamilton et al. 2005
Hamilton et al. 2005
Hamilton et al. 2005
Hamilton et al. 2005
Hamilton et al. 2005
Hamilton et al. 2005
Hamilton et al. 2005
Hamilton et al. 2005
a,b,c
a,b,c
a,b,c
a,b,c
a,b,c
a,b,c
a,b,c
a,b,c
a,b,c
a,b,c
Waddell and May 1995 a
Waddell and May 1995 a
Waddell and May 1995 a
50
25
C
'"egg-ovary
n
y
4
yv
o /o
o /
0 /
/
/ 0 v
0 />
CP P
muscle '"egg '"ovarv
6.30 6.50 7.00
15.60 46.50 30.60
29.20 37.80 45.50
5.10 6.00
5.80 - 5.90
13.50 - 27.50
16.20 - 42.10
6.00 - 5.10
12.50 - 10.00
18.00 - 12.90
4.40 3.70
7.10 4.70
32.00 10.60
Median ratio:
R2:
F:
df:
P:
^egg-ovary
6.75
38.55
41.65
6.00
5.90
27.50
42.10
5.10
10.00
12.90
3.70
4.70
10.60
1.12
0.61
12.5
8
0.004
Ratio
1.07
2.47
1.43
1.18
1.02
2.04
2.60
0.85
0.80
0.72
X
X
X
0 25 50
muscle
a Data from this study labeled above with 'x's' were excluded because results appeared atypical.
Do not distribute, quote or cite
B-31
Draft Document
-------
Roundtail chub (Gila robusta)
Study Cmuscie
Osmundson et al. 2007
Osmundson et al. 2007
Osmundson et al. 2007
Osmundson et al. 2007
Osmundson et al. 2007
Osmundson et al. 2007
Osmundson et al. 2007
30 -I
20 H
0 -O
C °e*
egg-ovary ^^~
10 J x«^0
o
01
^,,™,™,™,™,™,™,™,™,™,™,™,™,™,™,™
0 5 10 15
r
^muscle
\^ egg ^ ovarv ^ egg-ovary Iv3.ll 0
4.3 - 7.9 7.9 1.84
5 - 10.8 10.8 2.16
6.2 - 15.2 15.2 2.45
6.9 - 14.1 2.04
7 - 10.6 10.6 1.51
7.3 - 18 18 2.47
9.8 - 17.8 17.8 1.82
Median ratio: 2.04
R2: 0.62
F: 8.27
df: 5
P: 0.026
Smallmouth bass (Micropterus dolomieu)
Study Cmuscie
Osmundson et al. 2007
Osmundson et al. 2007
Osmundson et al. 2007
Osmundson et al. 2007
Osmundson et al. 2007
Osmundson et al. 2007
15 -I
10 rW11*****'^
'"egg-ovary Q^^Q
5 i
,,i, f! sj :
0 5 10 15
r
^muscle
\^ egg ^ ovarv ^ egg-ovarv Iv3ll 0
3.7 - 6.0 6.0 1.62
6.5 - 8.0 8.0 1.23
6.9 - 6.5 6.5 0.94
11 11 1.00
5.5 - 7.1 7.1 1.29
7.7 - 8.8 8.8 1.14
Median ratio: 1.19
R2: 0.85
F: 23.5
df: 4
P: 0.006
Do not distribute, quote or cite
B-32
Draft Document
-------
White Sturgeon (Acipenser transmontanus)
Study
Linville 2006
Linville 2006
Linville 2006
Linville 2006
Linville 2006
Linville 2006
20 •
c,^L'-
10 •
0
Cp
muscle ^es
1.28
1.22
1.48
9.93
15.3
11.1
O
, .&'
O
) 5 10 15 2Q
g *^ovarv
2.46
1.61
2.68
11
20.5
7.61
Median ratio:
R2:
F:
df:
P:
^egg-ovary
2.46
1.61
2.68
11
20.5
7.61
1.33
0.86
24.96
4
0.006
Ratio
2.46
1.61
2.68
11
20.5
7.61
White Sucker (Catostomus commersonii)
Study
Osmundson et al. 2007
Osmundson et al. 2007
Osmundson et al. 2007
Osmundson et al. 2007
Osmundson et al. 2007
Osmundson et al. 2007
Osmundson et al. 2007
Osmundson et al. 2007
Osmundson et al. 2007
Osmundson et al. 2007
Osmundson et al. 2007
Osmundson et al. 2007
Osmundson et al. 2007
Osmundson et al. 2007
Osmundson et al. 2007
Osmundson et al. 2007
Osmundson et al. 2007
Osmundson et al. 2007
Osmundson et al. 2007
Osmundson et al. 2007
Osmundson et al. 2007
Cp
muscle *^es
2.9
4.8
3.7
3.7
8.4
9.4
15.5
23.6
9.4
6.1
4.6
12.3
9.2
9.4
9.4
10.5
11.4
9.6
9.3
9.8
10.5
g ^ ovary
6.2
6.2
5.2
6.5
7.7
5.8
10.9
11.2
9.4
5.4
5.1
10.4
10.4
11
11.7
11.6
9.4
10.2
7.3
8.9
10.5
^egg-ovary
6.2
6.2
5.2
6.5
7.7
5.8
10.9
11.2
9.4
5.4
5.1
10.4
10.4
11
11.7
11.6
9.4
10.2
7.3
8.9
10.5
Ratio
2.14
1.29
1.41
1.76
0.92
0.62
0.70
0.47
1.00
0.89
1.11
0.85
1.13
1.17
1.24
1.10
0.82
1.06
0.78
0.91
1.00
Do not distribute, quote or cite
B-33
Draft Document
-------
White Sucker (Catostomus commersonii)
Study Cmuscie
Osmundson et al. 2007
Osmundson et al. 2007
Osmundson et al. 2007
Osmundson et al. 2007
Osmundson et al. 2007
Osmundson et al. 2007
Osmundson et al. 2007
Osmundson et al. 2007
Osmundson et al. 2007
Osmundson et al. 2007
Osmundson et al. 2007
Osmundson et al. 2007
Osmundson et al. 2007
Osmundson et al. 2007
Osmundson et al. 2007
Osmundson et al. 2007
Osmundson et al. 2007
Osmundson et al. 2007
Osmundson et al. 2007
20 -i
15"
^egg-ovary o3**n5r^
0.1 „„„„„„„„„,,, , ,„„„„„„„„„ , ., „„„„„„„„„„
j ™™™™™™™™.™ _, ,™™™™™™™™™ j j „„„„„„„„„»!
0 5 10 15 20 25
r
^ muscle
err
^egg ^ovarv ^^ egg-ovary
11.1 - 10.2 10.2
12.1 - 8.1 8.1
12.8 - 9.5 9.5
16.0 - 10.7 10.7
12.1 - 8.3 8.3
9.0 - 12 12
10.6 - 6.1 6.1
12.6 - 6.1 6.1
11.6 - 11.3 11.3
2.8 - 2.6 2.6
2.5 - 3.6 3.6
4.3 - 4.4 4.4
3.5 - 4.4 4.4
4.3 - 4.8 4.8
3.1 - 4.3 4.3
3.6 - 4.1 4.1
3.0 - 3.8 3.8
4.1 - 3.6 3.6
3.6 - 3.8 3.8
Median ratio: 1.00
R2: 0.59
F: 53.92
df: 38
P*" c\ nni
<- U.UUl
Ratio
0.92
0.67
0.74
0.67
0.69
1.33
0.58
0.48
0.97
0.93
1.44
1.02
1.26
1.12
1.39
1.14
1.27
0.88
1.06
Do not distribute, quote or cite
B-34
Draft Document
-------
Table B-3. Summary of muscle to egg-ovary conversion factors.
Common name
Bluegill
Bluehead sucker
Brook trout
Ccommon carp
Cutthroat trout
Dolly Varden
Flannelmouth sucker
Green sunfish
Mountain whitefish
Northern pike
Rainbow trout
Razorback sucker
Roundtail chub
Smallmouth bass
White sturgeon
White sucker
Scientific name
Lepomis macrochirus
Catostomus discobolus
Cyprinus carpio
Oncorhynchus clarkii
Catostomus latipinnis
Lepomis cyanellus
Oncorhynchus mykiss
Gila robusta
Micropterus dolomieu
Acipenser transmontanus
Catostomus commersonii
Median ratio
1.38
1.48
1.09
1.14
1.81
1.26
1.08
1.21
5.80
1.88
1.92
1.12
2.04
1.19
1.33
1.00
Do not distribute, quote or cite
B-35
Draft Document
-------
Muscle to whole-body conversion factors
Cwhoie-body = Selenium concentration
Cmusde = Selenium concentration
f
-p .• _ muscle
r
whole-body
in all tissues ((ig/g dw)
in muscle tissue only (|ig/g dw)
Black bullhead (Ameiurus melas)
Study
Osmundson et al. 2007
Osmundson et al. 2007
Osmundson et al. 2007
Osmundson et al. 2007
Osmundson et al. 2007
Osmundson et al. 2007
Osmundson et al. 2007
Osmundson et al. 2007
Osmundson et al. 2007
Osmundson et al. 2007
15 -|
10 -
o
r OQ, 0
5 °o
A
0 5 10
whole-body
^whole-bodv ^muscle JvatlO
5.30 3.40
4.80 3.90
5.50 4.30
4.90 4.70
9.60 5.70
7.60 7.40
7.30 7.50
6.60 7.80
8.60 7.80
2.00 9.20
Median ratio: 0.93
R2: 0.00
F: 0.03
df: 8
0.64
0.81
0.78
0.96
0.59
0.97
1.03
1.18
0.91
4.60
P: 0.973
15 Not used because P > 0.05.
Bluegill (Lepomis macrochirus)
Study
Doroshov et al. 1992
Doroshov et al. 1992
Doroshov et al. 1992
Doroshov et al. 1992
Hermanutz et al. 1996
Hermanutz et al. 1996
Hermanutz et al. 1996
Hermanutz et al. 1996
^whole-bodv ^muscle JvatlO
1.60 1.50
5.50 5.80
9.30 10.40
19.30 23.60
1.50 2.10
18.10 20.60
1.90 1.90
2.80 3.50
0.94
1.05
1.12
1.22
1.40
1.14
1.00
1.25
Do not distribute, quote or cite
B-36
Draft Document
-------
Bluegill (Lepomis macrochirus)
Study Cwhoie_bodv Cmuscie Ratio
Hermanutzetal. 1996 12.30 17.60
Hermanutzetal. 1996 9.40 12.50
Hermanutzetal. 1996 1.50 2.30
Hermanutz etal.1996 4.90 6.90
Hermanutzetal. 1996 21.00 44.90
Hermanutzetal. 1996 24.30 39.80
Hermanutzetal. 1996 2.70 3.40
Hermanutz etal.1996 5.00 5.30
Hermanutzetal. 1996 9.50 12.50
Hermanutzetal. 1996 6.60 7.80
Hermanutzetal. 1996 1.80 3.20
Hermanutzetal. 1996 4.20 6.10
Hermanutzetal. 1996 10.30 18.70
Hermanutzetal. 1996 13.80 15.10
Osmundson et al. 2007 8.80 12.90
50 -
o
C* 95
*-• muscle ZJ
0 -
(
X Median ratio:
R2:
F:
,f
df:
) 25 50
^-•whole-body
1.43
1.33
1.53
1.41
2.14
1.64
1.26
1.06
1.32
1.18
1.78
1.45
1.82
1.09
1.47
1.32
0.89
172.2
21
< 0.001
Do not distribute, quote or cite
B-37
Draft Document
-------
Bluehead sucker (Catostomus discobolus)
Study
Osmundson et al. 2007
Osmundson et al. 2007
Osmundson et al. 2007
Osmundson et al. 2007
Osmundson et al. 2007
Osmundson et al. 2007
Osmundson et al. 2007
^whole-bodv ^muscle JVallO
1.30
2.00
2.10
2.20
2.40
3.90
5.60
1.50
2.30
2.50
2.70
3.10
5.20
8.60
1.15
1.15
1.19
1.23
1.29
1.33
1.54
1U •
^muscle
X Median ratio: 1.23
R2: 0.99
F: 682.9
df: 5
P: <0.001
0 5 10
/-<
^whole-body
Brown trout (Salmo trutta)
Study Cwhoie.bodv Cmusde Ratio
Osmundson et al. 2007 4.60 3.20 0
Osmundson et al. 2007 4.30 3.60 0
Osmundson et al. 2007 5.00 4.00 0
Osmundson et al. 2007 5.50 6.30 1
70
84
80
15
-•muscle
4 -
4
Cwhole-b
Median ratio: 0.82
R2: 0.78
F: 7.2
df: 2
P: 0.122
Not used because P > 0.05.
Do not distribute, quote or cite
B-38
Draft Document
-------
Channel catfish (Ictalurus punctatus)
Study
Osmundson et al. 2007
Osmundson et al. 2007
Osmundson et al. 2007
Osmundson et al. 2007
^whole-bodv ^muscle JvatlO
3.40
3.30
2.60
4.00
3.40
3.60
3.70
5.30
1.00
1.09
1.42
1.33
b •
*-• muscle
0 -
C
0 Median ratio:
oX%^
i i Not use
3 6
/~<
^-•whole-body
R2
F:
df:
P:
d because P
1.21
: 0.49
2.0
2
0.338
>0.05.
Common carp (Cyprinus carpio)
Study Cwhoie.bodv
Osmundson et al. 2007 6.30
Osmundson et al. 2007 4.80
Osmundson et al. 2007 1 1 .70
Osmundson et al. 2007 23.10
Osmundson et al. 2007 4.10
/^
^^muscle
7.80
8.20
20.00
24.20
6.60
Ratio
1
1
1
1
1
24
71
71
05
61
-•muscle
30
20 -
10 -
Median ratio: 1.61
R2:
F:
df:
P:
0.85
17.6
3
0.017
10
20
30
-•whole-body
Do not distribute, quote or cite
B-39
Draft Document
-------
Flannelmouth sucker (Catostomus latipinnis)
Study
^whole-bodv ^muscle JVallO
Osmundson et al. 2007
Osmundson et al. 2007
Osmundson et al. 2007
Osmundson et al. 2007
Osmundson et al. 2007
Osmundson et al. 2007
Osmundson et al. 2007
10 -I
^muscle
0 -
(
0
*%
W
) 5 10
^whole body
3.0 3.6 1.20
2.6 3.8 1.46
3.1 4.1 1.32
3.1 4.6 1.48
3.5 5.2 1.49
4.4 6.2 1.41
4.5 7.3 1.62
Median ratio: 1.46
R2: 0.91
F: 50.1
df: 5
P: <0.001
Green sunfish (Lepomis cyanellus)
Study
^whole-bodv ^muscle KallO
Osmundson et al. 2007
Osmundson et al. 2007
Osmundson et al. 2007
Osmundson et al. 2007
Osmundson et al. 2007
Osmundson et al. 2007
Osmundson et al. 2007
Osmundson et al. 2007
Osmundson et al. 2007
Osmundson et al. 2007
Osmundson et al. 2007
Osmundson et al. 2007
Osmundson et al. 2007
Osmundson et al. 2007
Osmundson et al. 2007
Osmundson et al. 2007
Osmundson et al. 2007
Osmundson et al. 2007
Osmundson et al. 2007
Osmundson et al. 2007
22.80 28.10 1.23
8.80 12.90 1.47
15.40 21.90 1.42
4.80 5.00 1.04
5.70 6.10 1.07
4.40 5.20 1.18
3.80 5.10 1.34
11.90 15.70 1.32
6.40 10.10 1.58
9.50 11.50 1.21
9.10 10.50 1.15
6.20 7.20 1.16
7.00 9.30 1.33
7.70 7.70 1.00
6.20 6.00 0.97
10.20 12.00 1.18
9.70 12.10 1.25
9.90 12.50 1.26
7.20 7.50 1.04
9.00 11.30 1.26
Do not distribute, quote or cite
B-40
Draft Document
-------
Green sunfish (Lepomis cyanellus)
Study
Osmundson et al. 2007
Osmundson et al. 2007
Osmundson et al. 2007
Osmundson et al. 2007
Osmundson et al. 2007
Osmundson et al. 2007
Osmundson et al. 2007
Osmundson et al. 2007
Osmundson et al. 2007
Osmundson et al. 2007
Osmundson et al. 2007
Osmundson et al. 2007
Osmundson et al. 2007
Osmundson et al. 2007
Osmundson et al. 2007
Osmundson et al. 2007
Osmundson et al. 2007
Osmundson et al. 2007
Osmundson et al. 2007
Osmundson et al. 2007
Osmundson et al. 2007
Osmundson et al. 2007
Osmundson et al. 2007
Osmundson et al. 2007
Osmundson et al. 2007
Osmundson et al. 2007
Osmundson et al. 2007
Osmundson et al. 2007
Osmundson et al. 2007
Osmundson et al. 2007
Osmundson et al. 2007
Osmundson et al. 2007
Osmundson et al. 2007
whole-bodv
9.70
8.90
9.80
9.90
10.30
5.30
10.10
11.80
3.30
4.00
4.30
3.70
6.20
3.50
4.40
5.60
4.90
4.40
8.00
7.90
6.40
8.70
8.30
6.10
5.60
18.10
9.40
12.20
5.30
7.30
9.30
6.80
7.50
C muscle Ratio
13.60
13.20
12.40
12.50
8.60
5.30
11.90
13.60
3.80
4.20
4.10
4.20
5.70
4.40
3.50
5.50
5.00
4.30
10.10
11.90
11.10
11.80
11.00
7.10
6.70
26.40
9.60
16.70
8.10
10.60
14.20
11.30
12.80
1.40
1.48
1.27
1.26
0.83
1.00
1.18
1.15
1.15
1.05
0.95
1.14
0.92
1.26
0.80
0.98
1.02
0.98
1.26
1.51
1.73
1.36
1.33
1.16
1.20
1.46
1.02
1.37
1.53
1.45
1.53
1.66
1.71
Do not distribute, quote or cite
B-41
Draft Document
-------
Green sunfish (Lepomis cyanellus)
Study
C,
whole-body
c,
muscle
Ratio
30 -
C i 15 •
*-• muscle
0 -
C
Roundtail chub
Study
Osmundson et al
Osmundson et al
Osmundson et al
Osmundson et al
Osmundson et al
Osmundson et al
Osmundson et al
J/"
^?
S$s> °
#F
15
/~<
^-•whole-body
(Gila ro^wsta)
.2007
.2007
.2007
.2007
.2007
.2007
.2007
Median ratio:
30
whole-bodv
4.10
5.30
6.40
6.80
5.50
6.60
8.40
R2
F:
df:
P:
muscle
4.30
5.00
6.20
6.90
7.00
7.30
9.80
1.23
: 0.91
501.6
51
< 0.001
Ratio
1.05
0.94
0.97
1.01
1.27
1.11
1.17
10 n
-•muscle
5 -
Median ratio: 1.05
R2:
F:
df:
P:
10
0.86
29.6
5
0.002
^-•whole-body
Do not distribute, quote or cite
B-42
Draft Document
-------
Small mou th bass (Micropterus dolomieu)
Study
Osmundson
Osmundson
Osmundson
Osmundson
Osmundson
Osmundson
12 -I
^ muscle
0 -
et al. 2007
et al. 2007
et al. 2007
et al. 2007
et al. 2007
et al. 2007
whole-bodv
4.20
5.50
5.40
7.80
5.10
4.90
muscle
3.70
6.50
6.90
11.0
7.10
8.80
/ Median ratio:
o /
/°
f
o
0 6
whole-body
12
R2
F:
df:
P:
Ratio
0.88
1.18
1.28
1.41
1.08
1.57
1.23
: 0.83
20.2
4
0.008
White sucker (Catostomus commersonii)
Study
Osmundson
Osmundson
Osmundson
Osmundson
Osmundson
Osmundson
Osmundson
Osmundson
Osmundson
Osmundson
Osmundson
Osmundson
Osmundson
Osmundson
Osmundson
Osmundson
Osmundson
Osmundson
Osmundson
Osmundson
et al. 2007
et al. 2007
et al. 2007
et al. 2007
et al. 2007
et al. 2007
et al. 2007
et al. 2007
et al. 2007
et al. 2007
et al. 2007
et al. 2007
et al. 2007
et al. 2007
et al. 2007
et al. 2007
et al. 2007
et al. 2007
et al. 2007
et al. 2007
wh ol e-b odv
3.80
4.20
3.30
4.50
6.30
6.80
11.00
12.70
5.70
3.90
3.80
9.90
5.30
10.70
5.90
7.00
6.40
6.30
5.30
6.20
^muscle
2.90
4.80
3.70
3.70
8.40
9.40
15.50
23.60
9.40
6.10
4.60
12.30
9.20
9.40
9.40
10.50
11.40
9.60
9.30
9.80
Ratio
0.76
1.14
1.12
0.82
1.33
1.38
1.41
1.86
1.65
1.56
1.21
1.24
1.74
0.88
1.59
1.50
1.78
1.52
1.75
1.58
Do not distribute, quote or cite
B-43
Draft Document
-------
White sucker (Catostomus commersonii)
Study
Osmundson et al. 2007
Osmundson et al. 2007
Osmundson et al. 2007
Osmundson et al. 2007
Osmundson et al. 2007
Osmundson et al. 2007
Osmundson et al. 2007
Osmundson et al. 2007
Osmundson et al. 2007
Osmundson et al. 2007
Osmundson et al. 2007
Osmundson et al. 2007
Osmundson et al. 2007
Osmundson et al. 2007
Osmundson et al. 2007
Osmundson et al. 2007
Osmundson et al. 2007
Osmundson et al. 2007
Osmundson et al. 2007
Osmundson et al. 2007
Osmundson et al. 2007
Osmundson et al. 2007
Osmundson et al. 2007
Osmundson et al. 2007
Osmundson et al. 2007
Osmundson et al. 2007
Osmundson et al. 2007
Osmundson et al. 2007
Osmundson et al. 2007
Osmundson et al. 2007
Osmundson et al. 2007
Osmundson et al. 2007
Osmundson et al. 2007
Osmundson et al. 2007
Osmundson et al. 2007
Osmundson et al. 2007
Osmundson et al. 2007
Osmundson et al. 2007
Osmundson et al. 2007
whole-bodv
5.60
8.80
8.70
11.40
10.70
8.40
7.00
7.50
10.30
6.70
2.10
1.80
3.20
2.30
3.10
3.00
2.80
2.50
3.40
2.80
3.10
5.50
7.00
7.30
2.40
2.70
2.70
2.60
19.60
9.80
8.70
8.70
9.10
13.40
3.10
2.40
2.10
3.20
2.80
Cmuscie Ratio
10.50
11.10
12.10
12.80
16.00
12.10
9.00
10.60
12.60
11.60
2.80
2.50
4.30
3.50
4.30
3.10
3.60
3.00
4.10
3.60
5.60
6.30
9.10
8.50
3.00
4.40
3.20
1.60
28.10
12.10
11.80
12.60
12.30
18.00
2.80
3.20
3.10
4.30
3.40
1.88
1.26
1.39
1.12
1.50
1.44
1.29
1.41
1.22
1.73
1.33
1.39
1.34
1.52
1.39
1.03
1.29
1.20
1.21
1.29
1.81
1.15
1.30
1.16
1.25
1.63
1.19
0.62
1.43
1.23
1.36
1.45
1.35
1.34
0.90
1.33
1.48
1.34
1.21
Do not distribute, quote or cite
B-44
Draft Document
-------
White sucker (Catostomus commersonii)
Study
^whole-body ^muscle rvatlO
30 n
20 -
-• muscle
10 -
0 10 20
*-• whole -body
30
Median ratio: 1.34
R2:
F:
df:
P:
0.91
561.3
57
< 0.001
Table B-4. Muscle to whole-body correction factor.
Common name
Bluegill
Bluehead sucker
Common carp
Flannelmouth sucker
Green sunfish
Roundtail chub
Smallmouth bass
White sucker
Scientific name
Lepomis macrochirus
Catostomus discobolus
Cyprinus carpio
Catostomus latipinnis
Lepomis cyanellus
Gila robusta
Micropterus dolomieu
Catostomus commersonii
Median ratio
1.32
1.23
1.61
1.46
1.23
1.05
1.23
1.34
Table B-5. Directly calculated final whole-body to egg-ovary conversion factors (CF).
Common name
Median ratio
\\^ egg-ovary ^whole-body)
Median ratio
\\^ egg-ovary ^muscle)
Muscle to
whole-body
correction
factor
Final CF
values
Species
Bluegill
Bluehead sucker
Brook trout
Brown trout
Common carp
Cutthroat trout
2.13
1.82
1.45
1.92
1.96
1.09
1.27
2.13
1.82
1.38
1.45
1.92
1.96
Do not distribute, quote or cite
B-45
Draft Document
-------
Common name
Dolly varden
Flannelmouth sucker
Green sunfish
Mountain whitefish
Northern pike
Rainbow trout
Razorback sucker
Roundtail chub
Smallmouth bass
White sturgeon
White sucker
Median ratio
\ ess-ovary ^whole-bodv)
1.41
1.45
2.07
1.42
1.41
Median ratio
y^ ess-ovary ^muscle)
1.26
5.80
1.88
1.92
1.12
1.33
Muscle to
whole-body
correction
factor
1.27
1.27
1.27
1.27
1.34
1.27
Final CF
values
1.61
1.41
1.45
7.39
2.39
2.44
1.51
2.07
1.42
1.69
1.41
Genus
Catostomus
Esox
Lepomis
Micropterus
Oncorhynchus
1.41
2.39
1.79
1.42
1.96
Family
Catostomidae
Centrarchidae
Cyprinidae
Salmonidae
1.41
1.45
2.00
1.96
Order
Perciformes
1.45
Class
Actinopterygii
1.63
Do not distribute, quote or cite
B-46
Draft Document
-------
Table B-6. All EPA-derived egg-ovary to whole body (CF), egg-ovary to muscle, and muscle to whole body conversion factors directly calculated or estimated using taxonomic
classification (see main text for explanation of the taxonomic classification approach).
Common
name
Scientific name
Direct calculation
E-O/
WB
E-O/
M
M/
WB
Values based on taxonomic classification
Order
Family
Genus
E-O/
WB
E-O / WB
source
E-O/
M
E-O / M source
M/
WB
M / WB source
Final E-O /WB
Final
E-O
/WB
Final E-O / WB
source
alligator gar
bigmouth
buffalo
black
bullhead
black crappie
black
redhorse
blacknose
dace
blue catfish
bluegill
bluehead
sucker
brassy
minnow
brook
stickleback
brook trout
brown
bullhead
brown trout
brown
bullhead
bullhead
chain pickerel
channel
catfish
common carp
common
snook
crappie
creek chub
Atractosteus spatula
Ictiobus cyprinellus
Ameiurus melas
Pomoxis
nigromaculatus
Moxostoma
duquesnei
Rhinichthys
atratulus
Ictalurus furcatus
Lepomis
macrochirus
Catostomus
discobolus
Hybognathus
hankinsoni
Culaea inconstans
Salvelinus fontinalis
Ameiurus nebulosus
Salmo trutta
Ameiurus nebulosus
Esox
Ictalurus punctatus
Cyprinus carpio
Centropomus
undecimalis
Pomoxis sp.
Semotilus
atromaculatus
2.13
1.82
1.45
1.92
1.38
1.48
1.09
1.14
1.32
1.23
1.61
Lepistosteiformes
Cypriniformes
Siluriformes
Perciformes
Cypriniformes
Cypriniformes
Siluriformes
Perciformes
Cypriniformes
Cypriniformes
Gastero steiformes
Salmoniformes
Siluriformes
Salmoniformes
Siluriformes
Siluriformes
Esociformes
Siluriformes
Cypriniformes
Perciformes
Perciformes
Cypriniformes
Lepisosteidae
Catostomidae
Ictaluridae
Centrarchidae
Catostomidae
Cyprinidae
Ictaluridae
Centrarchidae
Catostomidae
Cyprinidae
Gastero steidae
Salmonidae
Ictaluridae
Salmonidae
Ictaluridae
Ictaluridae
Esocidae
Ictaluridae
Cyprinidae
Centropomidae
Centrarchidae
Cyprinidae
Atractosteus
Ictiobus
Ameiurus
Pomoxis
Moxostoma
Rhinichthys
Ictalurus
Lepomis
Catostomus
Hybognathus
Culaea
Salvelinus
Ameiurus
Salmo
Ameiurus
Esox
Ictalurus
Cyprinus
Centropomus
Pomoxis
Semotilus
1.63
1.41
1.63
1.45
1.41
2.00
1.63
2.13
1.82
2.00
1.63
1.71
1.63
1.45
1.63
1.63
1.63
1.63
1.92
1.45
1.45
2.00
All fish
Family
Catostomidae
All fish
Family
Centrarchidae
Family
Catostomidae
Family
Cyprinidae
All fish
Exact match
Exact match
Family
Cyprinidae
All fish
Family
Salmonidae
All fish
Exact match
All fish
All fish
All fish
All fish
Exact match
Order
Perciformes
Family
Centrarchidae
Family
Cyprinidae
1.30
1.10
1.30
1.21
1.10
1.59
1.30
1.38
1.48
1.59
1.30
1.09
1.30
1.81
1.30
1.30
1.88
1.30
1.14
1.21
1.21
1.59
All fish
Family
Catostomidae
All fish
Family
Centrarchidae
Family
Catostomidae
Family
Cyprinidae
All fish
Exact match
Exact match
Family
Cyprinidae
All fish
Exact match
All fish
Family
Salmonidae
All fish
All fish
Genus Esox
All fish
Exact match
Order
Perciformes
Family
Centrarchidae
Family
Cyprinidae
1.27
1.34
1.27
1.23
1.34
1.33
1.27
1.32
1.23
1.33
1.27
1.27
1.27
1.27
1.27
1.27
1.27
1.27
1.61
1.23
1.23
1.33
All fish
Family
Catostomidae
All fish
Family
Centrarchidae
Family
Catostomidae
Family
Cyprinidae
All fish
Exact match
Exact match
Family
Cyprinidae
All fish
All fish
All fish
All fish
All fish
All fish
All fish
All fish
Exact match
Order
Perciformes
Family
Centrarchidae
Family
Cyprinidae
1.63
1.41
1.63
1.45
1.41
2.00
1.63
2.13
1.82
2.00
1.63
1.38
1.63
1.45
1.63
1.63
2.39
1.63
1.92
1.45
1.45
2.00
All fish
Family
Catostomidae
All fish
Family
Centrarchidae
Family
Catostomidae
Family Cyprinidae
All fish
Exact match
Exact match
Family Cyprinidae
All fish
E-O/WB * M/WB
All fish
Exact match
All fish
All fish
E-O/WB * M/WB
All fish
Exact match
Order Perciformes
Family
Centrarchidae
Family Cyprinidae
Do not distribute, quote or cite
B-47
Draft Document
-------
Common
name
cutthroat trout
dolly varden
Scientific name
Oncorhynchus
clarkii
Salvelinus malma
fathead Pimephales
minnow promelas
flannelmouth
sucker
flathead
catfish
flathead chub
freshwater
drum
Catostomus
latipinnis
Pylodictis
Platygobio gracilis
Aplodinotus
grunniens
goldeye Hiodon alosoides
, , , Dorosoma
gizzard shad , .
0 cepedianum
green sunfish
iowa darter
Japanese
medaka
kokanee
salmon
largemouth
bass
largescale
sucker
Lepomis cyanellus
Etheostoma exile
Oryzias latipes
Oncorhynchus nerka
Micropterus
salmoides
Catostomus
macrocheilus
. , Rhinichthys
longnose dace , , J
0 cataractae
longnose
sucker
mosquito fish
mottled
sculpin
mountain
whitefish
northern pike
northern
pikeminnow
northern
plains killifish
Catostomus
catostomus
Gambusia sp.
Cottus bairdi
Prosopium
williamsoni
Esox lucius
Ptychocheilus
oregonensis
Fundulus kansae
Direct calculation
E-O/
WB
1.96
1.41
1.45
E-O/
M
1.81
1.26
1.08
1.21
5.80
1.88
M/
WB
1.46
1.23
Values based on taxonomic classification
Order
Salmoniformes
Salmoniformes
Cypriniformes
Cypriniformes
Siluriformes
Cypriniformes
Perciformes
Hiodontiformes
Clupeiformes
Perciformes
Perciformes
Beloniformes
Sahnoniformes
Perciformes
Cypriniformes
Cypriniformes
Cypriniformes
Cyprinodontiforme
s
Scorpaeniformes
Sahnoniformes
Esociformes
Cypriniformes
Cyprinodontiforme
s
Family
Salmonidae
Salmonidae
Cyprinidae
Catostomidae
Genus
Oncorhynchus
Salvelinus
Pimephales
Catostomus
Ictaluridae Pylodictus
Cyprinidae
Sciaenidae
Hiodontidae
Clupeidae
Centrarchidae
Percidae
Adrianichthyidae
Salmonidae
Centrarchidae
Catostomidae
Cyprinidae
Catostomidae
Poeciliidae
Cottidae
Salmonidae
Esocidae
Cyprinidae
Platygobio
Aplodinotus
Hiodon
Dorosoma
Lepomis
Etheostoma
Oryzias
Oncorhynchus
Micropterus
Catostomus
Rhinichthys
Catostomus
Gambusia
Cottus
Prosopium
Esox
Ptychocheilus
Fundulidae Fundulus
E-O/
WB
1.96
1.71
2.00
1.41
1.63
2.00
1.45
1.63
1.63
1.45
1.45
1.63
1.96
1.42
1.41
2.00
1.41
1.63
1.63
1.71
1.63
2.00
1.63
E-O / WB
source
Exact match
Family
Salmonidae
Family
Cyprinidae
Exact match
All fish
Family
Cyprinidae
Order
Perciformes
All fish
All fish
Exact match
Order
Perciformes
All fish
Genus
Oncorhynchus
Genus
Micropterus
Genus
Catostomus
Family
Cyprinidae
Genus
Catostomus
All fish
All fish
Family
Salmonidae
All fish
Family
Cyprinidae
All fish
E-O/
M
1.81
1.26
1.59
1.08
1.30
1.59
1.21
1.30
1.30
1.21
1.21
1.30
1.86
1.19
1.08
1.59
1.08
1.30
1.30
5.80
1.88
1.59
1.30
E-O / M source
Exact match
Exact match
Family
Cyprinidae
Exact match
All fish
Family
Cyprinidae
Order
Perciformes
All fish
All fish
Exact match
Order
Perciformes
All fish
Genus
Oncorhynchus
Genus
Micropterus
Genus
Catostomus
Family
Cyprinidae
Genus
Catostomus
All fish
All fish
Exact match
Exact match
Family
Cyprinidae
All fish
M/
WB
1.27
1.27
1.33
1.46
1.27
1.33
1.23
1.27
1.27
1.23
1.23
1.27
1.27
1.23
1.34
1.33
1.34
M / WB source
All fish
All fish
Family
Cyprinidae
Exact match
All fish
Family
Cyprinidae
Order
Perciformes
All fish
All fish
Exact match
Order
Perciformes
All fish
All fish
Genus
Micropterus
Genus
Catostomus
Family
Cyprinidae
Genus
Catostomus
1.27 All fish
1.27
1.27
1.27
1.33
1.27
All fish
All fish
All fish
Family
Cyprinidae
All fish
Final E-O /WB
Final
E-O
/WB
1.96
1.61
2.00
1.41
1.63
2.00
1.45
1.63
1.63
1.45
1.45
Final E-O / WB
source
Exact match
E-O/WB * M/WB
Family Cyprinidae
Exact match
All fish
Family Cyprinidae
Order Perciformes
All fish
All fish
Exact match
Order Perciformes
1.63 All fish
, „ , Genus
Oncorhynchus
1.42
1.41
2.00
1.41
Genus Micropterus
Genus Catostomus
Family Cyprinidae
Genus Catostomus
1.63 All fish
1.63
7.39
2.39
2.00
1.63
All fish
E-O/WB * M/WB
E-O/WB * M/WB
Family Cyprinidae
All fish
Do not distribute, quote or cite
B-48
Draft Document
-------
Common
name
northern
redbelly dace
northern
squawfish
quillback
rainbow trout
razorback
sucker
red shiner
redbreast
sunfish
redear sunfish
Scientific name
Chrosomus eos
Ptychocheilus
oregonensis
Carpiodes cyprinus
Oncorhynchus
mykiss
Xyrauchen texanus
Cyprinella lutrensis
Lepomis auritus
Lepomis
microlophus
... . . Richardsonius
redside shiner , . , ,
balteatus
river
carpsucker
river redhorse
rock bass
roundtail
chub
sacramento
perch
sacramento
pikeminnow
sailfin molly
sand shiner
sauger
sculpin
shadow bass
shorthead
redhorse
Carpiodes carpio
Moxostoma
carinatum
Ambloplites
rupestris
Gila robusta
Archoplites
interruptus
Ptychocheilus
grandis
Poecilia latipinna
Notropis stramineus
Sander canadensis
Cottus sp.
Ambloplites
ariommus
Moxostoma
macrolepidotum
.. Hypophthalmichthys
silver carp ,., .
^ mohtrix
Direct calculation
E-O/
WB
2.07
E-O/
M
1.92
1.12
2.04
M/
WB
1.05
Values based on taxonomic classification
Order
Cypriniformes
Cypriniformes
Cypriniformes
Salmoniformes
Cypriniformes
Cypriniformes
Perciformes
Perciformes
Cypriniformes
Cypriniformes
Cypriniformes
Perciformes
Cypriniformes
Perciformes
Cypriniformes
Cyprinodontiforme
s
Cypriniformes
Perciformes
Scorpaeniformes
Perciformes
Cypriniformes
Cypriniformes
Family
Cyprinidae
Cyprinidae
Catostomidae
Salmonidae
Catostomidae
Cyprinidae
Centrarchidae
Centrarchidae
Cyprinidae
Catostomidae
Catostomidae
Centrarchidae
Cyprinidae
Centrarchidae
Cyprinidae
Poeciliidae
Cyprinidae
Percidae
Cottidae
Centrarchidae
Catostomidae
Genus
Chrosomus
Ptychocheilus
Carpiodes
Oncorhynchus
Xyrauchen
Cyprinella
Lepomis
Lepomis
Richardsonius
Carpiodes
Moxostoma
Ambloplites
Gila
Archoplites
Ptychocheilus
Poecilia
Notropis
Sander
Cottus
Ambloplites
Moxostoma
„ ... Hypophthalmicht
Cyprinidae .
JF hys
E-O/
WB
2.00
2.00
1.41
1.96
1.41
2.00
1.79
1.79
2.00
1.41
1.41
1.45
2.07
1.45
2.00
1.63
2.00
1.45
1.63
1.45
1.41
2.00
E-O / WB
source
Family
Cyprinidae
Family
Cyprinidae
Family
Catostomidae
Genus
Oncorhynchus
Family
Catostomidae
Family
Cyprinidae
Genus Lepomis
E-O/
M
1.59
1.59
1.10
1.92
1.12
1.59
1.29
Genus Lepomis 1 .29
Family
Cyprinidae
Family
Catostomidae
Family
Catostomidae
Family
Centrarchidae
Exact match
Family
Centrarchidae
Family
Cyprinidae
All fish
Family
Cyprinidae
Order
Perciformes
All fish
Family
Centrarchidae
Family
Catostomidae
Family
Cyprinidae
1.59
1.10
1.10
1.21
2.04
1.21
1.59
1.30
1.59
1.21
1.30
1.21
1.10
1.59
E-O / M source
Family
Cyprinidae
Family
Cyprinidae
Family
Catostomidae
Exact match
Exact match
Family
Cyprinidae
Genus Lepomis
Genus Lepomis
Family
Cyprinidae
Family
Catostomidae
Family
Catostomidae
Family
Centrarchidae
Exact match
Family
Centrarchidae
Family
Cyprinidae
All fish
Family
Cyprinidae
Order
Perciformes
All fish
Family
Centrarchidae
Family
Catostomidae
Family
Cyprinidae
M/
WB
1.33
1.33
1.34
1.27
1.34
1.33
1.27
1.27
1.33
1.34
1.34
1.23
1.05
1.23
1.33
1.27
1.33
1.23
1.27
1.23
1.34
1.33
M / WB source
Family
Cyprinidae
Family
Cyprinidae
Family
Catostomidae
All fish
Family
Catostomidae
Family
Cyprinidae
Genus Lepomis
Genus Lepomis
Family
Cyprinidae
Family
Catostomidae
Family
Catostomidae
Family
Centrarchidae
Exact match
Family
Centrarchidae
Family
Cyprinidae
All fish
Family
Cyprinidae
Order
Perciformes
All fish
Family
Centrarchidae
Family
Catostomidae
Family
Cyprinidae
Final E-O /WB
Final
E-O
/WB
2.00
2.00
1.41
2.44
1.51
2.00
1.79
1.79
2.00
1.41
1.41
1.45
2.07
1.45
2.00
Final E-O / WB
source
Family Cyprinidae
Family Cyprinidae
Family
Catostomidae
E-O/WB * M/WB
E-O/WB * M/WB
Family Cyprinidae
Genus Lepomis
Genus Lepomis
Family Cyprinidae
Family
Catostomidae
Family
Catostomidae
Family
Centrarchidae
Exact match
Family
Centrarchidae
Family Cyprinidae
1.63 All fish
2.00
1.45
1.63
1.45
1.41
2.00
Family Cyprinidae
Order Perciformes
All fish
Family
Centrarchidae
Family
Catostomidae
Family Cyprinidae
Do not distribute, quote or cite
B-49
Draft Document
-------
Common
name
smallmouth
bass
smallmouth
buffalo
Scientific name
Micropterus
dolomieu
Ictiobus bubalus
speckled dace Rhinichthys osculus
spotted bass
spotted gar
stonecat
Micropterus
punctulatus
Lepisosteus oculatus
Noturus flavus
striped bass Morone saxatilis
striped mullet
sucker
tilapia
trout species
tui chub
utah sucker
walleye
western
mosquito fish
Mugil cephalus
Oncorhynchus sp.
Gila bicolor
Catostomus ardens
Sander vitreus
Gambusia affinis
westslope Oncorhynchus
cutthroat trout clarkii lewisi
white bass
white crappie
white
sturgeon
white sucker
wiper
yellow
bullhead
yellow perch
Morone chrysops
Pomoxis annularis
Acipenser
transmontanus
Catostomus
commersonii
Morone chrysops x
Moron saxatilis
Ameiurus natalis
Perca flavescens
Direct calculation
E-O/
WB
1.42
1.41
E-O/
M
1.19
1.33
1.00
M/
WB
1.23
1.34
Values based on taxonomic classification
Order
Perciformes
Cypriniformes
Cypriniformes
Perciformes
Lepistosteiformes
Siluriformes
Perciformes
Mugiliformes
Cypriniformes
Perciformes
Salmoniformes
Cypriniformes
Cypriniformes
Perciformes
Cyprinodontiforme
s
Salmoniformes
Perciformes
Perciformes
Acipenseriformes
Cypriniformes
Perciformes
Siluriformes
Perciformes
Family
Centrarchidae
Catostomidae
Cyprinidae
Centrarchidae
Lepisosteidae
Ictaluridae
Moronidae
Mugilidae
Catostomidae
Cichlidae
Salmonidae
Cyprinidae
Catostomidae
Percidae
Poeciliidae
Salmonidae
Moronidae
Centrarchidae
Acipenseridae
Catostomidae
Moronidae
Ictaluridae
Percidae
Genus
Micropterus
Ictiobus
Rhinichthys
Micropterus
Lepisosteus
Noturus
Morone
Mugil
Oncorhynchus
Gila
Catostomus
Sander
Gambusia
Oncorhynchus
Morone
Pomoxis
Acipenser
Catostomus
Morone
Ameiurus
Perca
E-O/
WB
1.42
1.41
2.00
1.42
1.63
1.63
1.45
1.63
1.41
1.45
1.96
2.07
1.41
1.45
1.63
1.96
1.45
1.45
1.63
1.41
1.45
1.63
1.45
E-O / WB
source
Exact match
Family
Catostomidae
Family
Cyprinidae
Genus
Micropterus
All fish
All fish
Order
Perciformes
All fish
Family
Catostomidae
Order
Perciformes
Genus
Oncorhynchus
Genus Gila
Genus
Catostomus
Order
Perciformes
All fish
Genus
Oncorhynchus
Order
Perciformes
Family
Centrarchidae
All fish
Exact match
Order
Perciformes
All fish
Order
Perciformes
E-O/
M
1.19
1.10
1.59
1.19
1.30
1.30
1.21
1.30
1.10
1.21
1.86
2.04
1.08
1.21
1.30
1.86
1.21
1.21
1.33
1.00
1.21
1.30
1.21
E-O / M source
Exact match
Family
Catostomidae
Family
Cyprinidae
Genus
Micropterus
All fish
All fish
Order
Perciformes
All fish
Family
Catostomidae
Order
Perciformes
Genus
Oncorhynchus
Genus Gila
Genus
Catostomus
Order
Perciformes
All fish
Genus
Oncorhynchus
Order
Perciformes
Family
Centrarchidae
Exact match
Exact match
Order
Perciformes
All fish
Order
Perciformes
M/
WB
1.23
1.34
1.33
1.23
1.27
1.27
1.23
1.27
1.34
1.23
1.27
1.05
1.34
1.23
1.27
1.27
1.23
1.23
1.27
1.34
1.23
M / WB source
Exact match
Family
Catostomidae
Family
Cyprinidae
Genus
Micropterus
All fish
All fish
Order
Perciformes
All fish
Family
Catostomidae
Order
Perciformes
All fish
Genus Gila
Genus
Catostomus
Order
Perciformes
All fish
All fish
Order
Perciformes
Family
Centrarchidae
All fish
Exact match
Order
Perciformes
1.27 All fish
1.23
Order
Perciformes
Final E-O /WB
Final
E-O
/WB
1.42
1.41
2.00
1.42
1.63
1.63
1.45
1.63
1.41
1.45
1.96
2.07
1.41
1.45
1.63
1.96
1.45
1.45
1.69
1.41
1.45
Final E-O / WB
source
Exact match
Family
Catostomidae
Family Cyprinidae
Genus Micropterus
All fish
All fish
Order Perciformes
All fish
Family
Catostomidae
Order Perciformes
Genus
Oncorhynchus
Genus Gila
Genus Catostomus
Order Perciformes
All fish
Genus
Oncorhynchus
Order Perciformes
Family
Centrarchidae
E-O/WB * M/WB
Exact match
Order Perciformes
1.63 All fish
1.45
Order Perciformes
Do not distribute, quote or cite
B-50
Draft Document
-------
DERIVATION OF TROPHIC TRANSFER FUNCTION VALUES
Methodology
Taxa specific trophic transfer functions (TTF) to quantify the degree of biomagnification across a
given trophic level were calculated from either physiological parameters measured in laboratory studies
or from field measurements of paired selenium concentrations in consumer species and their food. TTFs
from both approaches were used to calculate translated water concentrations; however, when TTF data of
similar quality are available from both approached, as was the case with bluegill, field-derived TTF data
are used.
Physiological data consisted of assimilation efficiencies (AE), measured as either a percentage or
a proportion, ingestion rates (IR), measured as grams of Se per grams of food consumed per day, and
efflux rate constant (ke), measured as I/day. All available data were collected for a particular species, and
then the TTF for that species was calculated using the equation:
Where AE, IR, and Ke were estimated as the median value of all available data for that parameter for that
species.
The majority of TTF were calculated using paired whole-body Se measurements from organisms
collected at the same site in the field. TTFs for trophic level 2 organisms were determined using the
equation:
rTL2
^tissue
' CTL2
Where Cj^2d equals the average Se concentration in particulate matter, defined as the average of Caigae,
Cdetntus, and CSediment. Of the three types of particulate matter potentially assumed by TL2 organisms (e.g.,
the majority of invertebrates), Csediment correlated relatively poorly to Q^fue, when compared to Caigae and
Cdetntus- In order to minimize potentially erroneous TTF calculations based solely on sediment Se
concentrations, while note completely discounting the importance of organic matter in sediments as a
potential food source, Csediment was included in CpartiCuiate calculations only when either Caigae or Cdetnms data
were also available.
Do not distribute, quote or cite B-51 Draft Document
-------
TTFs for trophic level 3 organisms were determined using the equation:
rTL3
tissue
Where Cj^d equals the average whole-body Se concentration in invertebrates collected at the same site
as their potential predator species. The majority of trophic level 3 organisms are fish species, but
damselflies and dragonflies of the order Odonata are also trophic level 3 organisms, and 7TFTL3 values
were calculated for those species as well.
For all field derived data used to determine TTFs, EPA first confirmed a statistical relationship
between whole-body selenium concentrations for each species and its food using OLS linear regression.
If the regression resulted in a statistically significant (P<0.05) positive slope, EPA calculated the TTF as
the median ratio of the paired concentration data.
Do not distribute, quote or cite B-52 Draft Document
-------
TTF values from physiological coefficients
AE (%) =
TTF
Assimilation efficiency
= Ingestion rate
= Efflux rate constant
AExlR
Invertebrates:
Baltic macoma (Macoma balthicd)
Physiological Parameters
AE (%) IR(g g1 d'1) ke (dj)
TTF Study
22.5
91.0
84.0
95.0
78.0
74.0
92.3
58.0
85.8
64.9
90.4
Median Values and TTF
84.0 0.27a
Luomaetal. 1992
Luomaetal. 1992
Luomaetal. 1992
Luomaetal. 1992
0.03 Reinfelderetal. 1997
0.03 Reinfelderetal. 1997
Schleckat et al. 2002
Schleckat et al. 2002
Schleckat et al. 2002
Schleckat et al. 2002
Schleckat et al. 2002
0.03 7.56
' Value taken from Mytilus edulls
Short-necked clam (Ruditapes philippinarum)
Physiological Parameters
AE (%) IR(g gj d'1) ke (dj)
TTF Study
70.0
52.0
Median Values and TTF
61.0 0.27a
0.013
0.013
0.013 12.67
Zhang etal. 1990
Zhang etal. 1990
Value taken from Mytilus edulis
Do not distribute, quote or cite
B-53
Draft Document
-------
Quahog (Mercenaria mercenarid)
Physiological Parameters
AE (%) IR(g g'1 d'1) ke (d'1)
TTF
Study
100.1
92.0
Median Values and TTF
96.1 0.27a
0.01
0.01 25.93
Reinfelder and Fisher
1994
Reinfelder et al. 1997
' Value taken from Mytilus edulis
Eastern Oyster (Crassostrea virginica)
Physiological Parameters
AE (%) IR(g g'1 d'1)
ke (d'1) TTF Study
105.4
70.0
Median Values and TTF
87.7 0.27a
0.005
0.070
0.038
Okazaki and Panietz
1981
Reinfelder and Fisher
1994
Reinfelder et al. 1997
6.31
' Value taken from Mytilus edulis
Common mussel (Mytilus edulis}
Physiological Parameters
AE(%) IRtgg'd1) ke(d1) TTF
86.0 0.02
75.0 0.05
60.7
48.0
13.7
55.1
55.8
71.9
71.5
27.9
84.4
81.0
79.4
Study
Reinfelder etal. 1997
Reinfelder etal. 1997
Wang and Fisher 1996
Wang and Fisher 1996
Wang and Fisher 1996
Wang and Fisher 1996
Wang and Fisher 1996
Wang and Fisher 1996
Wang and Fisher 1996
Wang and Fisher 1996
Wang and Fisher 1996
Wang and Fisher 1996
Wang and Fisher 1996
Do not distribute, quote or cite
B-54
Draft Document
-------
Physiological Parameters
AE (%) IR(g gj d'1)
TTF
Study
63.0
61.5
69.0
81.0
82.0
72.0
78.0
76.0
71.0
33.9
27.5
0.037
0.05
0.027
0.022
0.020
0.018
0.055
0.065
0.058
0.27 0.022
0.026
0.019
Wang and Fisher 1996
Wang and Fisher 1996
Wang and Fisher 1996
Wang and Fisher 1997
Wang and Fisher 1997
Wang and Fisher 1997
Wangetal. 1995
Wangetal. 1995
Wangetal. 1995
Wangetal. 1996
Wangetal. 1996
Wangetal. 1996
Wangetal. 1996
Wangetal. 1996
Wangetal. 1996
Wangetal. 1996
Median Values and TTF
71.3 0.27
0.026
7.30
Asian clam (Corbiculafluminea)
Physiological Parameters
AE (%) IR(g g'1 d'1)
TTF
Study
55.0 0.05 0.006
Median Values and TTF
55.0 0.05 0.006
Lee et al. 2006
4.58
Do not distribute, quote or cite
B-55
Draft Document
-------
Zebra mussel (Dreissenct polymorphd)
Physiological Parameters
AE (%) IR(g g'1 d'1) ke (d'1)
18.0
24.0
46.0
40.0
41.0
7.7
23.0
28.0
0.40
0.026
Median Values and TTF
26.0 0.40 0.026
TTF Study
Roditi and Fisher 1999
Roditi and Fisher 1999
Roditi and Fisher 1999
Roditi and Fisher 1999
Roditi and Fisher 1999
Roditi and Fisher 1999
Roditi and Fisher 1999
Roditi and Fisher 1999
Roditi and Fisher 1999
Roditi and Fisher 1999
4.00
Water flea (Daphnia magna)
Physiological Parameters
AE (%) IR(g g1 d'1) ke (dj)
0.08
0.34
57.9
43.0
39.8
33.0
41.4
41.5
38.0
24.5
0.101
0.12
0.131
0.134
0.108
0.112
Median Values and TTF
40.6 0.21 0.12
TTF Study
Goulet et al. 2007
Goulet et al. 2007
Yu and Wang 2002b
Yu and Wang 2002b
Yu and Wang 2002b
Yu and Wang 2002b
Yu and Wang 2002b
Yu and Wang 2002b
Yu and Wang 2002b
Yu and Wang 2002b
Yu and Wang 2002b
Yu and Wang 2002b
Yu and Wang 2002b
Yu and Wang 2002b
Yu and Wang 2002b
Yu and Wang 2002b
0.74
Do not distribute, quote or cite
B-56
Draft Document
-------
Copepod (Temora longicornis)
Physiological Parameters
AE (%) IR(g g'1 d'1) ke (d'1)
TTF
Study
55.0 0.42 0.115
Median Values and TTF
55.0 0.42 0.115
Wang and Fisher 1998
2.01
Copepod (Small, unidentified)
Physiological Parameters
AE (%) IR(g g'1 d'1)
ke (d'1) TTF Study
50.0 0.42
Median Values and TTF
50.0 0.42
0.155
0.155 1.35
Schlekat et al. 2004
Copepod (Large, unidentified)
Physiological Parameters
AE (%) IR(g gj d'1) ke
TTF Study
52.0
0.42
0.155
Schlekat et al. 2004
Median Values and TTF
50.0 0.42
0.155 1.41
Blackworm (Lumbriculus variegatus)
Physiological Parameters
AE (%) IR(g g'1 d'1)
TTF Study
24.0 0.067
9.0 0.067
Median Values and TTF
16.5 0.067
0.009
0.006
0.013
0.009
0.0086 1.29
Riedel and Cole 2001
Riedel and Cole 2001
Riedel and Cole 2001
Riedel and Cole 2001
Do not distribute, quote or cite
B-57
Draft Document
-------
Mayfly (Centroptilum triangulifeff
Physiological Parameters
AE (%) IR(g g'1 d'1)
38.0 0.72
40.0 0.72
Median Values and TTF
39.0 0.72
ke (d'1) TTF
0.25
0.19
0.22 1.28
Study
Riedel and
Riedel and
Cole 2001
Cole 2001
a - not used because field TTF data available
Vertebrates:
Bluegill (Lepomis macrochims)a
Physiological Parameters
AE(%) IRfeg-'d'1) ke(d1) TTF
34.0
22.0
24.0
36.0
30.0
32.0
43.0
40.0
37.0 0.041
0.031
0.034
36.0 0.031
0.038
0.038
0.008
0.042
Study
Besseretal. 1993
Besseretal. 1993
Besseretal. 1993
Besseretal. 1993
Besseretal. 1993
Besseretal. 1993
Besseretal. 1993
Besseretal. 1993
Besseretal. 1993
Besseretal. 1993
Besseretal. 1993
Besseretal. 1993
Besseretal. 1993
Besseretal. 1993
Whitledge and Haywood 2000
Whitledge and Haywood 2000
Median Values and TTF
35.0 0.025
0.036 1.1563
1 Not used because of availability of acceptable field-based TTF data
Do not distribute, quote or cite
B-58
Draft Document
-------
Fathead Minnow (Pimephales promelas)
Physiological Parameters
AE (%) IR(g g'1 d'1)
TTF Study
50.0
0.050
Median Values and TTF
50.0 0.050
0.029
0.019
0.3
0.014
0.013
0.016
0.012
0.026
0.018
0.025
0.0185
Presser and Luoma 2010
Bertram and Brooks 1986
Bertram and Brooks 1986
Bertram and Brooks 1986
Bertram and Brooks 1986
Bertram and Brooks 1986
Bertram and Brooks 1986
Bertram and Brooks 1986
Bertram and Brooks 1986
Bertram and Brooks 1986
Bertram and Brooks 1986
Bertram and Brooks 1986
1.35
Striped Bass (Morons saxatilis)
Physiological
AE (%) I]
33
42
Parameters
0.17
0.5
ke (d'1) TTF
0.09
0.08
Study
Baines et al. 2002
Baines et al. 2002
0.12
0.16
0.11
0.08
Median Values and TTF
37.5 0.335
Buckel and Stoner 2004
Buckel and Stoner 2004
Buckel and Stoner 2004
Buckel and Stoner 2004
0.085 1.48
Do not distribute, quote or cite
B-59
Draft Document
-------
TTF values from field
Invertebrates:
Calg =
c —
^ invert
Ratio
data
Selenium concentration in algae (mg/kg)
Selenium concentration in detritus (mg/kg)
Selenium concentration in sediment (mg/kg)
Selenium concentration in invertebrate tissue (mg/kg)
Average selenium concentration in particulate material (
('invert
('part
rca.lg+cdet+csed\
< 3
;
Scuds (Amphipoda)
Study
Birkner 1978
Birkner 1978
Birkner 1978
Birkner 1978
Birkner 1978
Birkner 1978
Birkner 1978
Birkner 1978
Lambing et al. 1994
Saikietal. 1993
Saikietal. 1993
Saikietal. 1993
Saikietal. 1993
Saikietal. 1993
Saikietal. 1993
Saikietal. 1993
Saikietal. 1993
Saikietal. 1993
Saikietal. 1993
Saikietal. 1993
Saikietal. 1993
Saikietal. 1993
Saikietal. 1993
Site
29
20
7
19
30
3
22
23
S46
ET6
ET6
GT5
GT5
GT4
GT4
SJR2
SJR2
SJR3
SJR3
SJR1
SJR1
ET7
ET7
Cal2
8.80
3.00
0.18
16.80
17.30
0.10
4.60
7.80
2.30
1.03
1.03
4.50
4.50
1.39
1.39
1.25
1.25
0.45
0.45
0.22
0.22
0.16
0.16
Cdet Csed '
15.40
41.00
2.80
1.20
47.30
0.30
44.00
10.80
1.15
1.15
14.95
14.95
8.40
8.40
5.00
5.00
1.25
1.25
0.50
0.50
0.76
0.76
-~i
12.10
22.00
1.49
9.00
32.30
0.20
24.30
9.30
2.30
1.09
1.09
9.73
9.73
4.90
4.90
3.13
3.13
0.85
0.85
0.36
0.36
0.46
0.46
^invert
18.40
11.40
2.90
4.30
22.50
2.30
7.60
11.30
3.20
0.44
0.86
4.60
3.30
3.40
3.70
3.80
2.80
1.50
1.10
0.89
1.30
1.10
1.10
Ratio
1.52
0.52
1.95
0.48
0.70
11.50
0.31
1.22
1.39
0.40
0.79
0.47
0.34
0.69
0.76
1.22
0.90
1.77
1.30
2.47
3.61
2.42
2.42
Do not distribute, quote or cite
B-60
Draft Document
-------
'inverts
Median ratio: 1.22
R2:
F:
df:
P:
0.69
46.9
21
< 0.001
Earthworms and Leeches (Annelida)
Study
Lemly 1985
Lemly 1985
Lemly 1985
Site
Badin Lake
Belews Lake
High Rock Lake
^ als ^ det ^ sed ^ part ^ invert -K3.il 0
8.20
62.70
8.25
0.91
8.27
0.79
4.56
35.49
4.52
8.10
51.15
9.05
1.78
1.44
2.00
'inverts
60 -
50 -
40 -
30 -
20 -
10 -
0
0
10
20
-•partic.
Median ratio: 1.78
R2:
F:
df:
P:
30
40
1.00
2426
1
< 0.001
Do not distribute, quote or cite
B-61
Draft Document
-------
Midges (Chironomidae)
Study
Birkner 1978
Birkner 1978
Birkner 1978
Birkner 1978
Birkner 1978
Birkner 1978
Birkner 1978
Birkner 1978
Grassoetal. 1995
Lambing etal. 1994
Saiki and Lowe 1987
Saiki and Lowe 1987
Saiki and Lowe 1987
Saiki and Lowe 1987
Saiki and Lowe 1987
Saiki and Lowe 1987
Saiki and Lowe 1987
Saiki and Lowe 1987
Saiki and Lowe 1987
Saiki and Lowe 1987
Saiki etal. 1993
Saiki etal. 1993
Saiki etal. 1993
Saiki etal. 1993
Saiki etal. 1993
Saiki etal. 1993
Saiki etal. 1993
Saiki etal. 1993
Saiki etal. 1993
Saiki etal. 1993
Saiki etal. 1993
Saiki etal. 1993
Saiki etal. 1993
Saiki etal. 1993
Site Calg Cdet Csed CDart (
29
19
30
3
22
27
12
23
17
S46
Kesterson Pond 1 1
Kesterson Pond 2
Kesterson Pond 2
Kesterson Pond 8
San Luis Drain
San Luis Drain
Volta Pond 26
Volta Pond 26
Volta Pond 7
Volta Pond 7
ET6
ET6
GT5
GT5
GT4
GT4
SJR2
SJR2
SJR3
SJR3
SJR1
SJR1
ET7
ET7
8.80
16.80
17.30
0.10
4.60
10.35
2.30
7.80
1.87
2.30
18.15
152.7
152.7
136.5
67.00
67.00
0.42
0.42
1.03
1.03
4.50
4.50
1.39
1.39
1.25
1.25
0.45
0.45
0.22
0.22
0.16
0.16
47.95
44.65
44.65
92.00
275.0
275.0
.01
.01
.39
.39
.15
.15
14.95
14.95
8.40
8.40
5.00
5.00
1.25
1.25
0.50
0.50
0.76
0.76
15.40
1.20
47.30
0.30
44.00
6.50
0.30
10.80
0.40
8.56
34.82
34.82
6.05
79.90
79.90
0.29
0.29
0.39
0.39
12.10
9.00
32.30
0.20
24.30
8.43
1.30
9.30
1.14
2.30
18.15
44.65
44.65
92.00
79.90
79.90
0.42
0.42
0.89
0.89
1.09
1.09
9.73
9.73
4.90
4.90
3.13
3.13
0.85
0.85
0.36
0.36
0.46
0.46
^invert Ratio
58.20
15.30
59.30
2.50
18.80
26.70
7.70
34.20
2.07
9.70
71.00
200.0
290.0
220.0
190.0
284.0
1.74
1.30
3.00
1.30
0.58
1.00
8.90
7.20
5.40
6.90
6.00
4.10
1.50
1.60
0.47
1.00
0.53
0.84
4.81
1.70
1.84
12.50
0.77
3.17
5.92
3.68
1.82
4.22
3.91
4.48
6.49
2.39
2.38
3.55
4.18
3.13
3.37
1.46
0.53
0.92
0.92
0.74
.10
.41
.92
.31
.77
.89
.31
2.78
.16
.85
Do not distribute, quote or cite
B-62
Draft Document
-------
Midges
Beetles
Study
Schuler
Schuler
Schuler
Schuler
Schuler
Schuler
Schuler
(
(Chiron omidae)
350 i
300
250
200
^-"inverts i CQ
100
° O /
/ 0
0 // o
50 -1 °y °
0 P^ °
0 50 100
C1
^-partic.
(Coleoptera)
Site Cal2 Cdet
etal. 1990 Kesterson 53.70
Pond 1 1
etal. 1990 Kesterson 53.70
Pond 11
etal. 1990 Kesterson 53.70
Pond 1 1
etal. 1990 Kesterson 52.50
Pond 2
etal. 1990 Kesterson 87.10
Pond 7
etal. 1990 Kesterson 87.10
Pond 7
etal. 1990 Kesterson 87.10
Pond 7
120 -i
100 -
80 -
60 -
'-inverts
40 -
20 -
ft -
0
0
-gl
^\,
o
o
o
Median ratio:
R2:
F:
df:
P:
Cp
sed t-Dart
11.50 32.60
11.50 32.60
11.50 32.60
9.30 30.90
5.90 46.50
5.90 46.50
5.90 46.50
Median ratio:
R2:
F:
df:
P:
Not used because P >
1.90
0.82
144.0
32
< 0.001
Cinvert Ratio
77.60 2
74.10 2
110.00 3
54.00 1
89.10 1
28.80 0
43.70 0
1.92
0.20
1.24
5
0.36
0.05 and negative
38
27
37
75
92
62
94
20
40
60
partic.
Do not distribute, quote or cite
B-63
Draft Document
-------
Water boatmen (Corixidae)
Study
Birkner 1978
Birkner 1978
Birkner 1978
Birkner 1978
Birkner 1978
Birkner 1978
Birkner 1978
Birkner 1978
Lambing et al. 1994
Rinellaetal. 1994
Rinellaetal. 1994
Rinellaetal. 1994
Saiki and Lowe 1987
Saiki and Lowe 1987
Saiki and Lowe 1987
Saiki and Lowe 1987
Saiki and Lowe 1987
Saiki and Lowe 1987
Saiki and Lowe 1987
Saiki and Lowe 1987
Schuleretal. 1990
Schuleretal. 1990
Schuleretal. 1990
Schuleretal. 1990
Schuleretal. 1990
Schuleretal. 1990
Schuleretal. 1990
Schuleretal. 1990
Rinella and Schuler
1992
Site
18
29
20
7
3
22
12
23
S46
G
A
Q
Kesterson Pond 1 1
Kesterson Pond 1 1
Kesterson Pond 8
Kesterson Pond 8
Volta Pond 26
Volta Pond 26
Volta Pond 7
Volta Pond 7
Kesterson Pond 1 1
Kesterson Pond 1 1
Kesterson Pond 1 1
Kesterson Pond 2
Kesterson Pond 2
Kesterson Pond 7
Kesterson Pond 7
Kesterson Pond 7
18
Calg <
7.60
8.80
3.00
0.18
0.10
4.60
2.30
7.80
2.30
0.84
2.21
1.42
18.15
18.15
136.50
136.50
0.42
0.42
53.70
53.70
53.70
52.50
52.50
87.10
87.10
87.10
0.59
^det ^^sed ^^part ^^ invert J\.«.11U
4.30
15.40
41.00
2.80
0.30
44.00
0.30
10.80
0.50
0.40
0.50
47.95 8.56
47.95 8.56
92.00 6.05
92.00 6.05
1.01 0.29
1.01 0.29
1.39 0.39
1.39 0.39
11.50
11.50
11.50
9.30
9.30
5.90
5.90
5.90
5.95
12.10
22.00
1.49
0.20
24.30
1.30
9.30
2.30
0.67
1.31
0.96
18.15
18.15
92.00
92.00
0.42
0.42
0.89
0.89
32.60
32.60
32.60
30.90
30.90
46.50
46.50
46.50
0.59
8.40
29.40
11.00
4.20
4.20
9.90
7.30
15.50
3.40
1.38
2.98
2.00
24.00
16.00
20.00
24.00
2.15
0.87
1.76
1.53
15.90
64.60
15.10
20.00
10.00
23.00
30.90
6.46
2.70
1.41
2.43
0.50
2.82
21.00
0.41
5.62
1.67
1.48
2.06
2.28
2.08
1.32
0.88
0.22
0.26
5.17
2.10
1.98
1.72
0.49
1.98
0.46
0.65
0.32
0.49
0.66
0.14
4.58
Do not distribute, quote or cite
B-64
Draft Document
-------
70 •
60 •
50 •
40 •
^-inverts go -
20 •
10 -
n j
o
o o
o o_V^~~^"
je?-5* o Q
0 50
'-partic.
o
Median ratio:
R2:
F:
df:
100
P:
1.48
0.25
9.17
27
< 0.001
Crayfish (Astacidae)
Study
Birkner 1978
Birkner 1978
Birkner 1978
Birkner 1978
Birkner 1978
Butler et al.
Butler et al.
Butler et al.
Butler et al.
Butler et al.
Butler et al.
Butler et al.
Butler et al.
Butler et al.
Butler et al.
Butler et al.
Butler et al.
Butler et al.
Butler et al.
Butler et al.
Butler et al.
Butler et al.
Butler et al.
Butler et al.
Butler et al.
Butler et al.
Butler et al.
Butler et al.
1993
1993
1995
1995
1995
1995
1995
1995
1995
1995
1995
1995
1995
1995
1995
1995
1995
1995
1995
1995
1995
1995
1997
Site
29
19
30
22
27
SP2
SP2
AK
AK
DD
DD
HD1
HD1
HD2
HD2
ME2
ME2
ME4
ME4
ME3
ME3
NW
NW
SD
SD
YJ2
YJ2
CHK
£
8
16
17
4
10
1
1
0
0
0
0
0
0
0
0
1
1
1
1
0
0
3
3
0
0
0
0
1
c c c
80
80
30
60
35
60
60
45
45
88
88
59
59
45
45
11
11
04
04
82
82
45
45
77
77
31
31
19
15
1
47
44
6
0
0
0
0
0
0
0
0
1
1
0
0
0
0
1
1
0
0
0
0
40
20
30
00
50
50
50
20
20
70
70
20
20
10
10
50
50
40
40
60
60
50
50
10
10
12
9
32
24
8
1
1
0
0
0
0
0
0
0
0
1
1
0
0
0
0
2
2
0
0
0
0
1
Cjnvert Ratio
10
00
30
30
43
05
05
33
33
79
79
59
59
32
32
10
10
77
77
61
61
53
53
64
64
21
21
19
23
10
36
11
20
2
2
0
0
0
1
0
0
0
3
4
3
1
1
1
1
0
30
10
80
30
00
60
90
76
79
62
10
86
79
96
00
10
40
30
80
40
70
20
30
40
40
40
50
90
1.93
1.12
1.14
0.47
2.37
2.48
2.76
2.34
2.43
0.78
1.39
1.46
1.34
2.98
3.10
1.00
1.27
1.69
2.35
2.30
6.07
1.66
1.31
2.20
2.20
6.83
7.32
0.76
Do not distribute, quote or cite
B-65
Draft Document
-------
Crayfish (Astacidae)
Butler etal. 1997 MN2
Butler etal. 1997 MUD2
Butler etal. 1997 MUD2
Butler etal. 1997 TRH
Butler etal. 1997 TRH
Saiki etal. 1993 ET6
Saiki etal. 1993 ET6
Saiki etal. 1993 GT5
Saiki etal. 1993 GT5
Saiki etal. 1993 GT4
Saiki etal. 1993 GT4
Saiki etal. 1993 SJR2
Saiki etal. 1993 SJR2
Saiki etal. 1993 SJR3
Saiki etal. 1993 SJR3
Saiki etal. 1993 SJR1
Saiki etal. 1993 SJR1
Saiki etal. 1993 ET7
Saiki etal. 1993 ET7
40 -
35 -
30 -
25 •
C „ 20 -
^ inverts
15 -
10 -
5 -
n J
0
1
1
1
1
1
1
4
4
1
1
1
1
0
0
0
0
0
0
79
30
30
25
25
03
03
50
50
39
39
25
25
45
45
22
22
16
16
o
1
1
14
14
8
8
5
5
1
1
0
0
0
0
15
15
95
95
40
40
00
00
25
25
50
50
76
76
0
9
9
4
4
3
3
0
0
0
0
0
0
.79
.30
.30
.25
.25
.09
.09
.73
.73
.90
.90
.13
.13
.85
.85
.36
.36
.46
.46
// Median ratio:
o
o /•
/
/r
/
^ R2:
o
F:
df:
P:
0
3
3
0
1
0
0
5
4
3
3
1
1
0
1
0
0
0
0
1.46
0.74
130.S
45
83
10
80
98
60
67
83
20
40
10
20
70
90
77
30
50
74
87
85
1
2
2
0
1
0
0
0
0
0
0
0
0
0
1
1
2
1
1
06
38
92
78
28
62
76
53
45
63
65
54
61
91
53
39
06
91
87
< 0.001
10 20 30
40
Do not distribute, quote or cite
B-66
Draft Document
-------
True flies (Diptera)
Study
Schuler et al.
Schuler et al.
Schuler et al.
Schuler et al.
Schuler et al.
Schuler et al.
Schuler et al.
Schuler et al.
Site
1990 Kesterson
1990 Kesterson
1990 Kesterson
1990 Kesterson
1990 Kesterson
1990 Kesterson
1990 Kesterson
1990 Kesterson
140 -
120 -
100 -
80 -
c
^ inverts gn .
40 -
20 -
n .
Calg
Pond
Pond
Pond
Pond
Pond
Pond
Pond
Pond
o
o
o~
o
o
11
11
2
2
2
7
7
7
~- — — —
53
53
52
52
52
87
87
87
-8
c
70
70
50
50
50
10
10
10
del Csed C
11.50
11.50
9.30
9.30
9.30
5.90
5.90
5.90
Median
part
32.60
32.60
30.90
30.90
30.90
46.50
46.50
46.50
ratio:
R2:
F:
df:
P:
Not used because P >
, slone.
Cjnvert Ratio
126
85
117
93
105
95
97
102
2.81
0.07
0.46
6
0.65
00
10
00
30
00
50
70
00
0.05 and nej
3.87
2.61
3.79
3.02
3.40
2.05
2.10
2.19
Dative
20
40
60
-partic.
Do not distribute, quote or cite
B-67
Draft Document
-------
Mayflies (Ephemeroptera)
Study
Rinella et al
Casey 2005
Casey 2005
Casey 2005
Casey 2005
Casey 2005
Casey 2005
Conley et al
Conley et al
Conley et al
Conley et al
Conley et al
Conley et al
Conley et al
Conley et al
Conley et al
Conley et al
Conley et al
Conley et al
Conley et al
Conley et al
Conley et al
*- inverts
Site Cal2 Cdet Csed CDart (
. 1994
.2009
.2009
.2009
.2009
.2009
.2009
.2009
.2011
.2011
.2011
.2013
.2013
.2013
.2013
.2013
90 -
80 -
70 -
60 -
50 -
40 -
30 -
20 -
10 -
n -
A
Deerlick Creek
Luscar Creek
Deerlick Creek
Luscar Creek
Deerlick Creek
Luscar Creek
Plate 10A
Plate 20A
Plate 20B
Plate 20C
Plate 20D
Plate 5A
Plate 5B
2x-High
2x-Low
2x-Medium
Control
Selenate-high
Selenate-low
Selenite-high
Selenite-low
o
_x"
s^ O
2
5
5
5
4
25
17
8
11
2
2
40
9
19
2
36
12
36
12
21
50
50
50
40
50
50
70
30
20
00
90
50
90
20
80
80
70
80
o
0.40 1
1.00 0.20 0
3.20 2.40 3
1.00 0.20 0
3.20 2.40 3
1.00 0.20 0
3.20 2.40 3
4
25
17
8
11
2
2
40
9
19
2
36
12
36
12
31
60
20
60
20
60
20
40
50
50
70
30
20
00
90
50
90
20
80
80
70
80
-invert Ratio
9
6
8
5
9
6
12
9
34
56
16
27
4
5
37
14
21
5
59
31
78
29
65
40
20
70
70
80
30
70
80
70
20
50
20
70
30
10
60
10
80
70
40
80
7
10
2
9
3
11
3
2
1
3
1
2
1
2
0
1
1
2
1
2
2
2
39
67
56
50
03
33
84
20
36
24
86
43
91
85
91
48
09
32
63
48
14
33
9-^ Median ratio: 2.38
o
R2: 0.75
F: 59.19
df: 20
P: <0.001
10
20 30
40
- participate
Do not distribute, quote or cite
B-68
Draft Document
-------
Snails (Gastropoda)
Study
Site Caig Cdet Csed
Butler etal. 1995 WC
Butler etal. 1995 WC
Butler etal. 1995 WC
Butler etal. 1997 DCP1
Butler etal. 1997 MNP2
Butler etal. 1997 CHP
Butler etal. 1997 LCHP1
20 -
15 -
f\ , 10 -
^•"inverts
5 -
n -
O
3.30 1
3.30 1
3.30 1
1.00 2
5.40 6
4.00 2
0.33 1
C
50
50
50
10
70
10
10
Median
part
2
2
2
1
6
3
0
.40
.40
.40
.55
.05
.05
.72
ratio:
R2:
F:
. . - df:
— o-~Q
o
o
024
'-partic.
° Not used because
6 8
P:
P>
^invert
3
3
2
3
2
19
0
1.54
0.01
0.07
5
0.93
0.05.
70
90
00
50
00
00
32
Ratio
1
1
0
2
0
6
0
54
63
83
26
33
23
45
Zooplankton
Study
Birkner 1978
Birkner 1978
Birkner 1978
Birkner 1978
Birkner 1978
Birkner 1978
Birkner 1978
Birkner 1978
Lambing et al.
Site
29
20
7
19
3
27
12
23
1988 12
Saiki and Lowe 1987 Kesterson
Pond 1 1
Saiki and Lowe 1987 Kesterson
Pond 2
Saiki and Lowe 1987 Kesterson
Pond8
Saiki and Lowe 1987 Volta Pond 26
Saiki and Lowe 1987 Volta Pond 7
Saiki and Lowe 1987 Volta
Wasteway
Saiki etal. 1993 ET6
CP P
alg ^det ^sed
8.80 15
3.00 41
0.18 2
16.80 1
0.10 0
10.35 6
2.30 0
7.80 10
1.40 0
18.15 47.95 8
152.70 44.65 34
136.50 92.00 6
0.42 1.01 0
1.39 0
0.87 2.03 0
1.03 1.15
C
40
00
80
20
30
50
30
80
30
56
82
05
29
39
24
part
12
22
1
9
0
8
1
9
0
18
44
92
0
0
0
1
.10
.00
.49
.00
.20
.43
.30
.30
.85
.15
.65
.00
.42
.89
.87
.09
^invert
31
11
3
7
3
42
5
15
2
68
30
00
30
70
40
50
80
40
60
30
83.00
100
1
2
2
1
00
46
90
80
20
Ratio
2
0
2
0
17
5
4
1
3
3
1
1
3
3
3
1
59
50
22
86
00
04
46
66
06
76
86
09
51
26
21
10
Do not distribute, quote or cite
B-69
Draft Document
-------
Zooplankton
Study
Site
Saikietal. 1993 ET6
Saikietal. 1993 GT5
Saikietal. 1993 GT5
Saikietal. 1993 GT4
Saikietal. 1993 GT4
Saikietal. 1993 SJR2
Saikietal. 1993 SJR2
Saikietal. 1993 SJR3
Saikietal. 1993 SJR3
Saikietal. 1993 SJR1
Saikietal. 1993 SJR1
Saikietal. 1993 ET7
Saikietal. 1993 ET7
140 -1
120 -
100 -
80 -
^inverts 6Q .
40 -
20 -
0 <
(
o °/
O s'
) 50
c
'"partic.
Ca,S
1
4
4
1
1
1
1
0
0
0
0
0
0
/
03
50
50
39
39
25
25
45
45
22
22
16
16
/
0
Cdet
1
14
14
CSed
15
95
95
8.40
8.40
5
5
1
1
0
0
0
0
100
00
00
25
25
50
50
76
76
cDart c
1.09
9.73
9.73
4.90
4.90
3.13
3.13
0.85
0.85
0.36
0.36
0.46
0.46
Median ratio:
R2
F:
df:
P:
invert Ratio
1.50
2.40
5.40
4.50
4.40
2.60
4.30
1.60
1.80
1.40
1.30
0.63
1.40
1.89
0.76
85.7
27
< 0.001
1.38
0.25
0.56
0.92
0.90
0.83
1.38
1.89
2.12
3.89
3.61
1.38
3.08
Do not distribute, quote or cite
B-70
Draft Document
-------
Special case of Odonates (Damselflies and Dragonflies) consuming invertebrates
Cfa
faod
damsel
Ratio
= Number of invertebrate food species co-occurring with an Odonate species.
= Average selenium concentration in particulate material (mg/kg):
fcalg+cdet+csed\
= Median selenium concentration in all invertebrate tissues that co-occur with an
Odonate species (mg/kg)
= Selenium concentration in damselfly tissue (mg/kg)
= Selenium concentration in dragonfly tissue (mg/kg)
_ Cfood
('part
cdamsel ^dragon
('food ('food
Co-occurring potential food species of damselflies and dragonflies (Odonata)
Study
Saiki and Lowe 1987
Saiki and Lowe 1987
Saiki and Lowe 1987
Saiki and Lowe 1987
Saiki and Lowe 1987
Saiki and Lowe 1987
Saiki and Lowe 1987
Saiki and Lowe 1987
Saiki and Lowe 1987
Saiki and Lowe 1987
Schuleretal. 1990
Schuleretal. 1990
Schuleretal. 1990
Schuleretal. 1990
Schuleretal. 1990
Schuleretal. 1990
Birkner 1978
Birkner 1978
Birkner 1978
Site
Kesterson Pond
11
Kesterson Pond
2
Kesterson Pond
2
Kesterson Pond
0
o
Kesterson Pond
0
o
Volta Pond 26
Volta Pond 26
Volta Pond 7
Volta Pond 7
Volta
Wasteway
Kesterson Pond
11
Kesterson Pond
11
Kesterson Pond
2
Kesterson Pond
2
Kesterson Pond
7
Kesterson Pond
7
29
20
7
Co-occurs with: n
dragonflies
dragonflies
dragonflies
dragonflies
dragonflies
dragonflies
dragonflies
dragonflies
dragonflies
dragonflies
dragonflies
dragonflies
dragonflies
dragonflies
dragonflies
dragonflies
damselflies
damselflies
damselflies
4
4
4
5
5
4
4
5
5
2
10
10
8
8
11
11
3
2
2
cpart
18.15
44.65
44.65
92.00
92.00
0.42
0.42
0.89
0.89
0.87
32.60
32.60
30.90
30.90
46.50
46.50
12.10
22.00
1.49
food
47.5
206.5
206.5
120
120
.52
.52
.53
.53
.83
75.85
75.85
93.3
93.3
69.2
69.2
29.4
11.2
3.55
Ratio
2.62
4.62
4.62
1.30
1.30
3.65
3.65
1.72
1.72
2.10
2.33
2.33
3.02
3.02
1.49
1.49
2.43
0.51
2.39
Do not distribute, quote or cite
B-71
Draft Document
-------
Co-occurring potential food species of damselflies and dragonflies (Odonatd)
Study
Birkner 1978
Birkner 1978
Birkner 1978
Birkner 1978
Birkner 1978
Birkner 1978
Site Co-occurs with: n
19 damselflies
30 damselflies
3 damselflies
22 damselflies
27 damselflies
23 damselflies
Grasso etal. 1995 17 damselflies
250 -
200 -
150 -
50 -
0 <
(
o
^^
^^^^ o
° ^^"^
) 20 40 60 80 100
*--particulate
CDart
2 9.00
2 32.30
3 0.20
3 24.30
1 8.43
3 9.30
1 1.14
Median ratio:
R2:
F:
df:
P:
Cfood Ratio
9.8
40.9
1
1
2.5 12
9.9
26.7
15.5
2.07
2.21
0.54
28.7
24
< 0.001
09
27
50
0.41
3
1
1
17
67
82
Damselflies (Anisoptera)
Study
Birkner 1978
Birkner 1978
Birkner 1978
Birkner 1978
Birkner 1978
Birkner 1978
Birkner 1978
Birkner 1978
Birkner 1978
Birkner 1978
Birkner 1978
Birkner 1978
Birkner 1978
Site Cfood
29
4
25
20
7
19
6
30
3
22
27
23
11
Grasso etal. 1995 17
Grasso etal. 1995 9
damsel
29.4
1.95
18.7
11.2
3.55
9.8
4.2
40.9
2.5
9.9
26.7
15.5
5.9
2.07
8.2
Ratio
55
1.8
21.9
18.7
4.4
28.4
11.1
53.3
3.1
15.8
45.1
18.4
7.7
1.75
6.98
1
0
1
1
1
2
2
1
1
1
1
1
1
0
0
87
92
17
67
24
90
64
30
24
60
69
19
31
85
85
Do not distribute, quote or cite
B-72
Draft Document
-------
Damselflies (Anisopterd)
Study
100 -I
80 -
60 -
'-damsel
40 -
20 -
0 -
(
Site
/
Cfood
Median
Cdamsel Ratio
ratio: 1.30x2.21
/^ (damselfly food to particulate) = 2.88
o /\-»
* /*
ep-o°
) 20
•^ ^ u
40 60
od
R2: 0.89
F: 104.4
df: 13
P: <0.001
Dragonflies (Zygoptera)
Study
Mason et al. 2000
Mason et al. 2000
Saiki and Lowe 1987
Saiki and Lowe 1987
Saiki and Lowe 1987
Saiki and Lowe 1987
Saiki and Lowe 1987
Saiki and Lowe 1987
Saiki and Lowe 1987
Saiki and Lowe 1987
Saiki and Lowe 1987
Saiki and Lowe 1987
Schuleretal. 1990
Schuleretal. 1990
Schuleretal. 1990
Schuleretal. 1990
Schuleretal. 1990
Schuleretal. 1990
Sorenson & Schwarzbach 1991
Site
BK
HCRT
Kesterson Pond 1 1
Kesterson Pond 2
Kesterson Pond 2
Kesterson Pond 8
Kesterson Pond 8
Volta Pond 26
Volta Pond 26
Volta Pond 7
Volta Pond 7
Volta Wasteway
Kesterson Pond 1 1
Kesterson Pond 1 1
Kesterson Pond 2
Kesterson Pond 2
Kesterson Pond 7
Kesterson Pond 7
5
t^food
1.845
4.305
47.5
206.5
206.5
120
120
1.52
1.52
1.53
1.53
1.83
75.85
75.85
93.3
93.3
69.2
69.2
0.42
Cdragon Ratio
1.665
2.81
53
155
171
95.5
105
1.4
1.42
1.2
1.4
2.5
63.1
95.5
110
65
61.7
56.2
0.49
0.90
0.65
1.12
0.75
0.83
0.80
0.88
0.92
0.93
0.78
0.92
1.37
0.83
1.26
1.18
0.70
0.89
0.81
1.17
Do not distribute, quote or cite
B-73
Draft Document
-------
Dragonflies (Zygoptera)
Study
200 -I
150 -
r 100 -
'-'dragon
50 •
n i
Site
/°
/°
Cp
food ^dragon
Median ratio:
Ratio
0.89x2.21
/ (damselfly food to particulate) = 1 .97
O o/
o J$
„ flp°
Q>/*3
/
•
R2:
F:
df:
P:
0.95
343.5
17
<0.001
0 100 200 300
Vertebrates:
c
^ invert
Cfish
T?atir»
IxailO
Cfood
= Selenium concentration in invertebrate tissue ((ig/g)
= Average selenium concentration
_ cfish
^invert
in the whole-body offish ((ig/g)
Black bullhead (Ameiurus melas)
Study
GEI2013
GEI2013
GEI2013
GEI2013
GEI2013
Lemly 1985
Site
Sand Creek at Colfax
Sand Creek at Colfax
Sand Creek at Colfax
Sand Creek at Colfax
Sand Creek at Colfax
Badin Lake
Mueller et al. 1991 Lake Meredith near Ordway,
CO
Mueller et al. 1991 Lake Meredith near Ordway,
Lemly 1985
CO
High Rock Lake
Mueller et al. 1991 Pueblo Reservoir near Pueblo,
CO
Butler etal. 1991 SweitzerLake
Lemly 1985
Belews Lake
Cr<
invert *^fish
2.81
2.81
2.81
2.81
2.81
5.18
6.40
6.40
6.75
8.70
29.80
45.53
Ratio
1.95
2.37
2.73
3.21
3.96
4.19
9.20
9.70
5.26
7.40
39.00
28.11
0.70
0.84
0.97
1.14
1.41
0.81
1.44
1.52
0.78
0.85
1.31
0.62
Do not distribute, quote or cite
B-74
Draft Document
-------
Black bullhead (Ameiurus melas)
Study
50 -
40 -
30 -
Cflsh 2Q _
10 - CU-""
n ™
0 10
Site Cjnvert Cflsh
o
^^ Median ratio:
^^"^ o
^^ R2.
*^ F:
df:
p.
20 30 40 50
^- Invert.
Ratio
0.91
0.79
38.1
10
< 0.001
Black crappie (Pomoxis nigromaculatus)
Study
Butler etal. 1995
Butler etal. 1995
Peterson et al. 1991
Peterson et al. 1991
Mueller etal. 1991
Lambing etal. 1994
Lambing etal. 1994
Lambing etal. 1994
Lambing etal. 1994
Lambing etal. 1994
Lambing etal. 1994
Site Cjnvert Cflsh
Totten Reservoir 1.07
Summit Re servoir 1.85
Ocean Lake, west side 3.83
Ocean Lake, west side 3.83
Lake Meredith near 6.40
Ordway, CO
Priest Butte Lakes near 14.00
Choteau
Priest Butte Lakes near 14.00
Choteau
Priest Butte Lakes near 14.00
Choteau
Priest Butte Lakes near 15.00
Choteau
Priest Butte Lakes near 15.00
Choteau
Priest Butte Lakes near 15.00
Choteau
Ratio
2.50
1.70
4.20
6.32
13.00
39.00
41.00
47.00
40.00
57.00
63.00
2.35
0.92
1.10
1.65
2.03
2.79
2.93
3.36
2.67
3.80
4.20
Do not distribute, quote or cite
B-75
Draft Document
-------
Black crappie (Pomoxis nigromaculatus)
Study
Site
- invert
cfl;
ish
Ratio
70 -I
60 -
50 -
., 40 -
-fish 30 .
20 -
10 -
0
o
o
Median ratio: 2.67
0
10
15
20
R2:
F:
df:
P:
0.92
97.9
9
< 0.001
-invert.
Blacknose dace (Rhinichthys atratulus)
Study
Mason et al. 2000
Mason et al. 2000
Mason et al. 2000
Site
BK
BK
BK
Cr<
invert *^fish
1.43
1.43
1.43
Ratio
1.13
1.45
1.74
0.79
1.01
1.21
o
o
o
Median ratio: 1.01
R2:
F:
df:
P:
0.0
0.0
1
1.0
Not used because P > 0.05.
C
*" invert.
Bluegill (Lepomis macrochirus)
Study
Saikietal. 1993
Saikietal. 1993
Saikietal. 1993
Saikietal. 1993
Site
ET6
ET6
ET7
ET7
CP
invert ^fist
0.85
0.85
0.86
0.86
, Ratio
1.40
2.20
1.20
1.20
1.66
2.60
1.40
1.40
Do not distribute, quote or cite
B-76
Draft Document
-------
Bluegill (Lepomis macrochirus)
Study
Hermanutz et al. 1996
Saikietal. 1993
Saikietal. 1993
Butler etal. 1995
Hermanutz et al. 1996
Saikietal. 1993
Saikietal. 1993
Hermanutz et al. 1996
Saikietal. 1993
Saikietal. 1993
Hermanutz et al. 1996
Saikietal. 1993
Saikietal. 1993
Saikietal. 1993
Saikietal. 1993
Hermanutz et al. 1996
Hermanutz et al. 1996
Hermanutz et al. 1996
Crutchfield 2000
Crutchfield 2000
Crutchfield 2000
Crutchfield 2000
Mueller etal. 1991
Crutchfield 2000
Crutchfield 2000
Crutchfield 2000
Crutchfield 2000
Hermanutz et al. 1996
Crutchfield 2000
Crutchfield 2000
Crutchfield 2000
Crutchfield 2000
Crutchfield 2000
Crutchfield 2000
Crutchfield 2000
Crutchfield 2000
Crutchfield 2000
Crutchfield 2000
Crutchfield 2000
Crutchfield 2000
Hermanutz et al. 1996
Crutchfield 2000
Site
MSO I
SJR1
SJR1
TT
MSO III
SJR3
SJR3
MSO II
SJR2
SJR2
MSO III
GT4
GT4
GT5
GT5
MSO II
MSO II
MSO III
transect 3
transect 3
transect 3
transect 3
Rl
transect 3
transect 3
transect 3
transect 3
MSO III
transect 3
transect 3
transect 3
transect 3
transect 3
transect 3
transect 4
transect 4
transect 4
transect 4
transect 4
transect 4
MSO II
transect 3
P P
^ invert ^fis
0.87
0.95
0.95
1.07
1.20
1.50
1.50
1.70
3.30
3.30
3.95
4.05
4.05
4.90
4.90
5.05
5.05
5.55
8.60
8.60
8.60
8.60
8.70
9.25
9.25
9.25
9.25
10.00
11.40
11.40
11.95
11.95
15.20
15.20
15.70
15.70
15.70
15.70
16.45
16.45
16.63
16.95
;h Ratio
1.55
0.87
1.40
2.30
1.83
1.90
2.00
1.55
2.70
3.30
4.21
4.30
4.50
5.00
6.40
3.86
4.88
13.77
7.64
7.64
11.90
14.30
5.20
7.64
9.05
16.70
19.00
10.32
30.32
34.50
21.28
28.60
38.73
48.80
15.16
16.70
20.20
21.28
13.10
13.63
24.29
16.70
1.78
0.92
1.48
2.16
1.52
1.27
1.33
0.91
0.82
1.00
1.06
1.06
1.11
1.02
1.31
0.76
0.97
2.48
0.89
0.89
1.38
1.66
0.60
0.83
0.98
1.81
2.05
1.03
2.66
3.03
1.78
2.39
2.55
3.21
0.97
1.06
1.29
1.36
0.80
0.83
1.46
0.99
Do not distribute, quote or cite
B-77
Draft Document
-------
Bluegill (Lepomis macrochirus)
Study
Crutchfield 2000
Crutchfield 2000
Crutchfield 2000
Hermanutz et al. 1996
Hermanutz et al. 1996
Crutchfield 2000
Crutchfield 2000
Crutchfield 2000
Crutchfield 2000
Crutchfield 2000
Crutchfield 2000
Crutchfield 2000
Crutchfield 2000
Crutchfield 2000
Crutchfield 2000
Crutchfield 2000
Crutchfield 2000
Hermanutz et al. 1996
Hermanutz et al. 1996
Crutchfield 2000
Crutchfield 2000
Crutchfield 2000
Crutchfield 2000
Crutchfield 2000
Crutchfield 2000
Crutchfield 2000
Crutchfield 2000
Crutchfield 2000
Crutchfield 2000
Crutchfield 2000
Crutchfield 2000
Crutchfield 2000
Crutchfield 2000
Crutchfield 2000
Crutchfield 2000
Crutchfield 2000
Crutchfield 2000
Crutchfield 2000
Crutchfield 2000
Crutchfield 2000
Crutchfield 2000
Crutchfield 2000
Site
transect 3
transect 3
transect 3
MSOII
MSOII
transect 3
transect 3
transect 4
transect 4
transect 3
transect 3
transect 3
transect 3
transect 4
transect 4
transect 4
transect 4
MSOI
MSOI
transect 3
transect 3
transect 3
transect 3
transect 3
transect 3
transect 4
transect 4
transect 4
transect 4
transect 4
transect 4
transect 4
transect 4
transect 4
transect 4
transect 4
transect 4
transect 4
transect 4
transect 4
transect 4
transect 4
P P
^ invert ^fi
16.95
16.95
16.95
17.30
17.30
17.90
17.90
18.25
18.25
20.35
20.35
20.70
20.70
20.90
20.90
20.90
20.90
21.19
21.19
21.80
21.80
21.80
21.80
23.40
23.40
25.40
25.40
30.00
30.00
30.70
30.70
33.20
33.20
33.25
33.25
38.55
38.55
43.90
43.90
48.90
48.90
49.30
ish Ratio
33.38
38.10
48.54
16.76
20.99
33.38
44.00
16.69
20.20
50.07
83.30
27.27
41.70
27.27
39.30
43.96
52.40
18.13
18.28
26.20
28.03
33.38
65.50
46.25
84.50
30.32
45.20
51.60
60.70
86.51
102.40
53.13
59.50
54.66
61.90
83.46
237.00
37.97
48.80
62.18
156.00
72.75
1.97
2.25
2.86
0.97
1.21
1.86
2.46
0.91
1.11
2.46
4.09
1.32
2.01
1.30
1.88
2.10
2.51
0.86
0.86
1.20
1.29
1.53
3.00
1.98
3.61
1.19
1.78
1.72
2.02
2.82
3.34
1.60
1.79
1.64
1.86
2.16
6.15
0.86
1.11
1.27
3.19
1.48
Do not distribute, quote or cite
B-78
Draft Document
-------
Bluegill (Lepomis macrochirus)
Study
Crutchfield 2000
250 -|
200 -
150 -
Cfish 100 -
50 -
o -nd
0
Bluehead sucker
Study
Butler etal. 1995
Butler etal. 1995
Butler etal. 1995
Butler etal. 1995
Butler etal. 1995
Butler etal. 1995
Butler etal. 1995
Butler etal. 1995
Butler etal. 1993
Butler etal. 1993
Butler etal. 1993
Butler etal. 1995
Butler etal. 1995
Butler etal. 1993
Butler etal. 1995
Butler etal. 1995
Butler etal. 1993
Butler etal. 1993
Butler etal. 1993
Butler etal. 1994
Butler etal. 1994
Butler etal. 1995
Butler etal. 1995
Butler etal. 1994
Butler etal. 1997
Site ^invert
Cf^ Ratio
transect 4 49.30 92.90 1.88
o
Median ratio: 1.48
o
0
O O O J3H--*"""""'/^
JD ..Q-ff"""""^ a
eJtifg""'6^ 8
ftfipW*
20 40 60
(Catostomus discobolus)
Site {^invert
R2: 0.58
F: 119.4
df: 87
P: < 0.001
Cflsh Ratio
AK 0.78 0.94 1.21
HD1 0.83 0.83 1.01
HD1 0.83 0.86 1.04
HD1 0.83 1.20 1.45
HD1 0.83 1.40 1.70
DD 0.86 0.64 0.74
DD 0.86 0.88 1.02
DD 0.86 1.30 1.51
Dl
Bl
Bl
ME2
ME2
B2
SD
SD
D2
D2
PI
COL1
RB3
YJ2
YJ2
.20 2.80 2.33
.25 1.90 1.52
.25 2.20 1.76
.25 0.83 0.66
.25 1.30 1.04
.35 1.80 1.33
.40 1.50 1.07
.40 1.80 1.29
.45 1.60 1.10
.45 2.30 1.59
.50 2.20 1.47
.50 1.60 1.07
.60 13.00 8.13
.65 0.96 0.58
.65 2.80 1.70
NFK3 2.00 1.40 0.70
MN2 2.20 1.20 0.55
Do not distribute, quote or cite
B-79
Draft Document
-------
Bluehead sucker
Study
Butler etal. 1997
Butler etal. 1997
Butler etal. 1997
Butler etal. 1997
Butler etal. 1993
Butler etal. 1995
Butler etal. 1995
Butler etal. 1995
Butler etal. 1995
Butler etal. 1995
Butler etal. 1997
Butler etal. 1997
Butler etal. 1993
Butler etal. 1993
Butler etal. 1997
Butler etal. 1997
Butler etal. 1997
Butler etal. 1993
Butler etal. 1991
Butler etal. 1993
Butler etal. 1994
Butler etal. 1997
Butler etal. 1997
Butler etal. 1995
Butler etal. 1995
Butler etal. 1994
Butler etal. 1994
Butler etal. 1994
35 -
30 -
25 -
c 20 "
o
10 -
5 - _flf
n -^Hl
U 1
0
(Catostomus discobolus)
Site
MUD
MUD
CHK
CHK
Ul
SJ1
SJ1
SJ1
ME3
ME3
MN3
MN1
SP1
SP2
MUD2
MUD2
MUD2
F2
4
F2
BSW1
WBR
WBR
NW
NW
LZA1
RBI
GUN2
o
o
op _^~ -— - -6~~~~~~"
\ o^
$8
10 20
^-Invert.
C C
^invert ^fish
2.30
2.30
2.40
2.40
2.45
2.50
2.50
2.50
2.55
2.55
2.70
2.90
2.95
3.40
3.45
3.45
3.45
3.90
3.90
4.80
5.00
5.05
5.05
5.10
5.10
19.00
21.00
28.00
Median ratio:
_- R2:
F:
df:
P
30
Ratio
1.80
2.30
1.20
1.60
4.80
0.94
1.20
1.20
1.70
1.80
1.50
1.40
5.10
7.10
2.50
5.20
5.60
10.00
1.80
0.94
33.00
1.80
2.80
7.20
9.30
9.00
22.00
3.60
1.04
0.16
9.6
51
< 0.001
0.78
1.00
0.50
0.67
1.96
0.38
0.48
0.48
0.67
0.71
0.56
0.48
1.73
2.09
0.72
1.51
1.62
2.56
0.46
0.20
6.60
0.36
0.55
1.41
1.82
0.47
1.05
0.13
Do not distribute, quote or cite
B-80
Draft Document
-------
Brook stickleback (Culaea inconstans)
Study
GEI2013
GEI2013
GEI2013
GEI2013
GEI2013
GEI2013
GEI2013
GEI2013
GEI2013
GEI2013
GEI2013
GEI2013
GEI2013
Lambing et al. 1994
Lambing et al. 1994
Lambing et al. 1994
GEI2013
GEI2013
GEI2013
GEI2013
GEI2013
GEI2013
GEI2013
GEI2013
GEI2013
GEI2013
GEI2013
GEI2013
GEI2013
GEI2013
GEI2013
GEI2013
GEI2013
GEI2013
GEI2013
Lambing et al. 1994
Lambing et al. 1994
Lambing et al. 1994
Site
SWA1
SWA1
SWA1
SWA1
SWA1
SWA1
SWA1
SWA1
SWA1
SWA1
SWA1
SWA1
SWA1
S38
S37
S36
SW2-1
SW2-1
SW2-1
SW2-1
SW2-1
SW2-1
SW2-1
SW2-1
SW2-1
SW2-1
SWB
SWB
SW1
SW1
SW1
SW1
SW11
SW11
SW2-1
S34
Sll
Sll
P P
^ invert ^fis
2.81
2.81
2.81
2.81
2.81
3.64
3.64
3.64
3.64
3.64
3.64
3.64
3.64
4.70
5.30
6.30
6.60
6.60
6.60
6.60
6.60
6.60
6.60
6.60
6.60
6.60
7.06
7.06
7.82
7.82
7.82
7.82
8.41
8.41
9.14
14.00
14.50
14.50
;h Ratio
4.40
4.59
4.66
5.00
5.21
3.69
4.16
4.21
4.62
4.78
4.98
5.06
6.28
17.00
6.10
5.30
21.14
23.21
23.64
25.89
27.71
32.97
34.54
37.05
39.26
43.38
15.74
17.15
9.96
10.38
10.58
11.98
6.36
6.45
21.09
35.00
22.00
26.00
1.57
1.64
1.66
1.78
1.86
1.02
1.14
1.16
1.27
1.31
1.37
1.39
1.73
3.62
1.15
0.84
3.21
3.52
3.58
3.93
4.20
5.00
5.24
5.62
5.95
6.58
2.23
2.43
1.27
1.33
1.35
1.53
0.76
0.77
2.31
2.50
1.52
1.79
Do not distribute, quote or cite
B-81
Draft Document
-------
Brook stickleback (Culaea inconstans)
Study
50 -
40 -
30 -
Cfish
o
10 - ^^
08 °
A
0 5
Site
o
8^-""°
8
o o
10 15 20
^-invert.
Cjnvert Cflsh Ratio
Median ratio: 1.69
R2: 0.27
F: 13.3
df: 36
P: < 0.001
Brook trout (Salvelinus fontinalis)
Study
Hamilton and Buhl 2004
Mason et al. 2000
Mason et al. 2000
Mason et al. 2000
Mason et al. 2000
Mason et al. 2000
Mason et al. 2000
Butler etal. 1997
Hamilton and Buhl 2005
Hamilton and Buhl 2005
Hamilton and Buhl 2004
Hamilton and Buhl 2004
12 -
10 -
8 -
Cfish 6 •
'"o J^
2 - °JK#
T
0
Site
use
BK
BK
BK
HCRT
HCRT
HCRT
MN1
LGC
UGC
DVC
use
o .,
o^^^o
5 10 15
^- invert.
Cjnvert Cf,sh Ratio
0.50 2.40
1.43 1.21
1.43 1.57
1.43 1.90
2.81 0.99
2.81 1.59
2.81 2.95
2.90 2.20
7.80 6.90
9.30 9.80
12.80 8.00
0.50 2.40
Median ratio: 0.88
R2: 0.83
F: 43.6
df: 9
P: < 0.001
4.80
0.84
1.10
1.33
0.35
0.57
1.05
0.76
0.88
1.05
0.63
4.80
Do not distribute, quote or cite
B-82
Draft Document
-------
Brown bullhead (Ameiurus nebulosus)
Study
Rinella and Schuler 1992
Mason et al. 2000
Mason et al. 2000
Mason et al. 2000
21 0,
—
Cfish ! -
1 -
1 I
0 1
Site
HCRT
HCRT
HCRT
\.
^\
2
c c
^invert ^fish
1.20
2.81
2.81
2.81
o
o Median ratio:
R2:
F:
o df:
1 P:
Not used because P >
slope
1.90
0.22
1.23
1.83
0.55
0.27
0.73
Ratio
1.58
0.08
0.44
0.65
2
0.58
0.05 and negative
Brown trout (Salmo truttd)
Study
Butler etal. 1993
Butler etal. 1993
Butler etal. 1993
Butler etal. 1993
Butler etal. 1993
Butler etal. 1993
Butler etal. 1993
Butler etal. 1993
Butler etal. 1993
Butler etal. 1993
Butler etal. 1993
Butler etal. 1993
Formation 2012
Butler etal. 1993
Butler etal. 1994
Formation 2012
Formation 2012
Butler etal. 1993
Butler etal. 1991
Formation 2012
Site
LP2
LP2
LP2
LP3
LP3
Bl
B2
B2
B2
D2
D2
D2
SFTC-1
PI
NFK3
SFTC-1
SFTC-1
SP2
12
CC-75
Cp
invert ^fish
1.00
1.00
1.00
1.12
1.12
1.25
1.35
1.35
1.35
1.45
1.45
1.45
1.63
1.95
2.00
2.42
2.49
2.75
2.80
3.11
1.60
1.70
2.10
2.10
2.80
4.20
2.40
2.70
2.70
3.20
3.50
3.50
6.70
3.30
5.00
3.68
2.64
1.20
5.40
4.05
Ratio
1.60
1.70
2.10
1.88
2.51
3.36
1.78
2.00
2.00
2.21
2.41
2.41
4.11
1.69
2.50
1.52
1.06
0.44
1.93
1.30
Do not distribute, quote or cite
B-83
Draft Document
-------
Brown trout (Salmo trutta)
Study
Formation 2012
Formation 2012
Formation 2012
Butler etal. 1993
Formation 2012
Butler etal. 1993
Butler etal. 1993
Butler etal. 1993
Butler etal. 1993
Butler etal. 1991
Butler etal. 1991
Formation 2012
Butler etal. 1993
Formation 2012
Formation 2012
Formation 2012
McDonald and Strosher 1998
Formation 2012
Formation 2012
Formation 2012
Butler etal. 1991
Butler etal. 1994
Butler etal. 1994
Butler etal. 1994
Butler etal. 1994
Formation 2012
Formation 2012
Formation 2012
Butler etal. 1991
Hamilton and Buhl 2005
Formation 2012
Formation 2012
Formation 2012
Formation 2012
Formation 2012
Formation 2012
Formation 2012
Formation 2012
Formation 2012
Formation 2012
Formation 2012
Formation 2012
Site
CC-75
CC-350
CC-350
LP4
SFTC-1
SP2
R2
R2
R2
4
4
CC-75
ST2
CC-75
CC-75
CC-350
ER747
CC-150
CC-150
CC-150
10
SMF
SMF
SMF
SMF
CC-3A
CC-3A
CC-3A
3
CC
CC-150
DC-600
DC-600
DC-600
DC-600
DC-600
LSV-4
LSV-4
HS-3
HS-3
CC-350
CC-350
c c
^^ invert ^fis
3.11
3.16
3.16
3.20
3.21
3.40
3.70
3.90
3.90
3.90
3.90
3.97
4.10
4.16
4.16
4.20
4.29
4.46
4.46
4.70
4.80
4.80
4.80
4.80
4.80
5.45
5.45
5.48
6.20
6.70
7.03
7.83
7.83
8.53
8.53
8.65
9.54
9.54
11.40
11.40
11.45
11.45
;h Ratio
5.35
6.28
8.53
1.80
2.25
3.40
5.90
5.40
6.70
3.30
3.50
3.18
6.00
6.60
10.32
5.78
4.80
5.83
8.67
5.20
2.00
8.40
8.54
9.40
21.44
9.20
10.44
11.25
3.50
9.70
10.14
10.54
12.83
6.20
8.54
5.85
15.18
16.20
18.83
20.60
7.95
11.50
1.72
1.99
2.70
0.56
0.70
1.00
1.59
1.38
1.72
0.85
0.90
0.80
1.46
1.59
2.48
1.38
1.12
1.31
1.94
1.11
0.42
1.75
1.78
1.96
4.47
1.69
1.92
2.05
0.56
1.45
1.44
1.35
1.64
0.73
1.00
0.68
1.59
1.70
1.65
1.81
0.69
1.00
Do not distribute, quote or cite
B-84
Draft Document
-------
Brown trout (Salmo trutta)
Study
Formation 2012
Formation 2012
Formation 2012
Formation 2012
Formation 2012
Formation 2012
Formation 2012
Formation 2012
Formation 2012
Formation 2012
Formation 2012
Formation 2012
Butler etal. 1994
Butler etal. 1994
Formation 2012
Formation 2012
Formation 2012
Formation 2012
Formation 2012
Formation 2012
Formation 2012
Formation 2012
Butler etal. 1994
Butler etal. 1994
Formation 2012
100 -|
80 -
60 -
Cfisl1 40 -
20 -
n -
o os,j
i§i@r®
Site
CC-1A
CC-1A
CC-1A
CC-1A
HS-3
CC-1A
CC-150
CC-3A
CC-3A
HS
HS
HS
HCC1
HCC1
LSV-2C
LSV-2C
HS-3
LSV-2C
HS-3
LSV-2C
HS
HS
GUN2
GUN2
LSV-2C
o
8
ntJL-ogg'"
O o
p
^ invert
12
12
12
12
Cfish
24
24
24
57
13.41
13
14
14
14
15
15
18
21
21
22
22
24
26
26
26
27
27
28
28
30
55
32
50
50
70
70
70
00
00
62
62
70
31
55
95
80
80
00
00
00
Median ratio:
R2:
F:
df:
P:
Ratio
9.33
10.51
16.85
9.95
17.89
14.03
7.83
15.38
19.68
16.52
25.00
24.90
35.68
42.00
12.78
19.45
23.68
22.67
28.97
20.96
22.80
32.63
5.90
80.27
19.53
1.44
0.55
102.7
85
< 0.001
0.76
0.86
1.38
0.79
1.33
1.04
0.55
1.06
1.36
1.05
1.59
1.33
1.70
2.00
0.56
0.86
0.96
0.86
1.09
0.78
0.82
1.17
0.21
2.87
0.65
10
20 30
40
Do not distribute, quote or cite
B-85
Draft Document
-------
Bullhead (Ameiurus sp.)
Study
Butler etal. 1995
Butler etal. 1993
Butler etal. 1993
Butler etal. 1994
5 -,
4 -
3 -
Cfish -,
1 -
A
0
Site
ME3
R2
R2
BSW1
o^^-<
cr-^^
1 '
2 4
*- Invert,
C C
*-* invert ^fish
2.55
3.70
3.70
5.00
3 Median ratio:
R2:
F:
df:
P:
Not used because P >
6
Ratio
3.00
3.50
4.00
4.10
1.01
0.77
6.58
2
0.13
0.05
1.18
0.95
1.08
0.82
Channel catfish (Ictalurus punctatus)
Study
Butler etal. 1995
Butler etal. 1995
Butler etal. 1995
Butler etal. 1997
Roddy etal. 1991
Roddy etal. 1991
Roddy etal. 1991
Roddy etal. 1991
Roddy etal. 1991
Roddy etal. 1991
Roddy etal. 1991
Roddy etal. 1991
Roddy etal. 1991
Roddy etal. 1991
Roddy etal. 1991
Roddy etal. 1991
Roddy etal. 1991
Roddy etal. 1991
Roddy etal. 1991
Butler etal. 1993
Butler etal. 1993
Butler etal. 1993
Site
TT
SJ1
SJ1
MN4
18
18
18
18
18
18
18
18
18
18
18
18
18
18
18
LP4
LP4
R2
P P
^^ invert ^fish
1.07
2.50
2.50
2.65
3.10
3.10
3.10
3.10
3.10
3.10
3.10
3.10
3.10
3.10
3.10
3.10
3.10
3.10
3.10
3.20
3.20
3.70
Ratio
1.00
2.80
4.10
4.20
.40
.50
.60
.70
.70
.80
.80
.90
2.00
2.10
2.20
2.20
2.30
2.40
3.10
2.68
3.30
3.00
0.94
1.12
1.64
1.58
0.45
0.48
0.52
0.55
0.55
0.58
0.58
0.61
0.65
0.68
0.71
0.71
0.74
0.77
1.00
0.84
1.03
0.81
Do not distribute, quote or cite
B-86
Draft Document
-------
Channel catfish (Ictalurus punctatus)
Study
Butler etal. 1993
Butler etal. 1993
Butler etal. 1993
Butler etal. 1997
Mueller etal. 1991
Butler etal. 1991
Butler etal. 1991
Butler etal. 1991
Butler etal. 1991
Butler etal. 1991
Butler etal. 1991
Butler etal. 1991
50 -
40 -
30 -
Cfisa 2Q _
10 • o ^"
^J
-------
Common carp (Cyprinus carpio)
Study
Butler etal. 1995
Garcia-Hernandez et al. 2000
Butler etal. 1994
Roddy etal. 1991
Roddy etal. 1991
Roddy etal. 1991
Roddy etal. 1991
Roddy etal. 1991
Roddy etal. 1991
GEI2013
GEI2013
GEI2013
GEI2013
GEI2013
GEI2013
GEI2013
Butler etal. 1993
Peterson et al. 1991
Peterson et al. 1991
Peterson et al. 1991
Peterson et al. 1991
Peterson et al. 1991
Butler etal. 1993
GEI2013
GEI2013
GEI2013
GEI2013
Butler etal. 1991
Butler etal. 1993
GEI2013
GEI2013
GEI2013
GEI2013
GEI2013
GEI2013
GEI2013
GEI2013
GEI2013
GEI2013
GEI2013
GEI2013
GEI2013
Site
MN1
NFK2
18
18
18
18
18
18
SW4-
SW4-
SW4-
SW4-
SW4-
SW4-
SW4-
R2
7
7
7
7
7
R2
SW88
SW88
SW88
SW88
9
R2
SW9
SW9
SW9
SW9
SW9
SW9
SW9
SW9
SW9
SW9
SW9
SW9
SW9
CP
invert ^fisl
2.70
3.00
3.10
3.10
3.10
3.10
3.10
3.10
3.10
3.33
3.33
3.33
3.33
3.33
3.33
3.33
3.70
3.83
3.83
3.83
3.83
3.83
3.90
3.96
3.96
3.96
3.96
4.10
4.30
4.45
4.45
4.45
4.45
4.45
4.45
4.45
4.45
4.45
4.45
4.45
4.45
4.45
, Ratio
9.80
3.30
4.90
3.20
3.90
4.60
4.70
4.80
5.30
3.91
4.36
4.48
4.60
4.78
7.29
9.61
3.30
4.24
4.41
4.73
5.16
5.21
4.80
3.88
5.33
5.49
5.66
3.90
5.00
3.64
3.70
3.77
3.80
3.90
4.14
4.26
4.41
4.50
4.53
4.69
5.61
6.13
3.63
1.10
1.58
1.03
1.26
1.48
1.52
1.55
1.71
1.18
1.31
1.35
1.38
1.44
2.19
2.89
0.89
1.11
1.15
1.23
1.35
1.36
1.23
0.98
1.35
1.39
1.43
0.95
1.16
0.82
0.83
0.85
0.85
0.88
0.93
0.96
0.99
1.01
1.02
1.05
1.26
1.38
Do not distribute, quote or cite
B-88
Draft Document
-------
Common carp (Cyprinus carpio)
Study
Butler etal. 1991
Butler etal. 1994
Lemly 1985
Low and Mullins 1990
Mueller etal. 1991
Mueller etal. 1991
Butler etal. 1991
Mueller etal. 1991
Mueller etal. 1991
GEI2013
GEI2013
GEI2013
GEI2013
GEI2013
GEI2013
GEI2013
GEI2013
GEI2013
GEI2013
Lemly 1985
GEI2013
GEI2013
Butler etal. 1993
Grassoetal. 1995
Grassoetal. 1995
Grassoetal. 1995
May et al. 2008
May et al. 2008
GEI2013
GEI2013
GEI2013
GEI2013
GEI2013
GEI2013
GEI2013
GEI2013
GEI2013
GEI2013
GEI2013
Mueller etal. 1991
Butler etal. 1997
Mueller etal. 1991
Site
10
BSW1
Badin lake
5
A6
A3
3
R2
R2
SW2-
SW2-
SW2-
SW2-
SW2-
SW2-
SW2-
SW2-
SW2-
SW2-
High Rock Lake
SWB
SWB
F2
9
9
9
SSW
SSAU
SW11
SW11
SW11
SW11
SW11
SW11
SW11
SW11
SW11
SW11
SW11
A2
MN5
Rl
c c
^^ invert ^fis
4.80
5.00
5.18
5.60
5.60
6.00
6.20
6.40
6.40
6.60
6.60
6.60
6.60
6.60
6.60
6.60
6.60
6.60
6.60
6.75
7.06
7.06
7.50
7.59
7.59
7.59
7.60
8.35
8.41
8.41
8.41
8.41
8.41
8.41
8.41
8.41
8.41
8.41
8.41
8.50
8.60
8.70
h Ratio
10.30
12.00
6.50
1.20
3.40
6.50
2.20
14.00
14.40
22.96
24.27
25.09
26.73
26.74
28.74
29.73
31.74
36.81
41.57
5.03
12.50
15.61
5.80
4.70
4.93
5.51
10.40
7.59
3.56
3.60
3.79
3.95
4.14
4.34
6.43
7.21
7.50
7.90
11.84
7.30
16.00
5.60
2.15
2.40
1.26
0.21
0.61
1.08
0.35
2.19
2.25
3.48
3.68
3.80
4.05
4.05
4.36
4.51
4.81
5.58
6.30
0.75
1.77
2.21
0.77
0.62
0.65
0.73
1.37
0.91
0.42
0.43
0.45
0.47
0.49
0.52
0.77
0.86
0.89
0.94
1.41
0.86
1.86
0.64
Do not distribute, quote or cite
B-89
Draft Document
-------
Common carp (Cyprinus carpio)
Study
May et al. 2008
GEI2013
GEI2013
GEI2013
GEI2013
GEI2013
GEI2013
May et al. 2008
May et al. 2008
May et al. 2008
Lambing etal. 1994
Lambing etal. 1994
May et al. 2008
Butler etal. 1994
May et al. 2008
Butler etal. 1994
Butler etal. 1991
Butler etal. 1991
Butler etal. 1991
Butler etal. 1991
Lemly 1985
70
60
50
40 0
CO
Fisli „ Q O
20 O Q^"
££^C
10 (efiyaftr) „
o I&^S^
Site
NSK
SW2-
SW2-
SW2-
SW2-
SW2-
SW2-
sso
NSCL
SSAL
S34
S34
KR
RBI
NSP
GUN2
7
7
7
7
Belews Lake
o
0 ^^°
/v
0
o
o
0 10 20 30 40 50 60
Creek chub (Semotilus
Study
Mason et al. 2000
Mason et al. 2000
Mason et al. 2000
GEI2013
^Invert
atromaculatus)
Site
HCRT
HCRT
HCRT
SW4-1
c c
^invert ^fish
8.81
9.14
9.14
9.14
9.14
9.14
9.14
10.00
10.70
11.50
14.00
14.00
17.20
21.00
24.00
28.00
29.80
29.80
29.80
29.80
45.53
Median ratio:
R2:
F:
df:
P:
C C
^invert ^fish
2.81
2.81
2.81
3.33
Ratio
9.33
13.29
13.77
20.49
23.65
24.84
27.27
8.48
10.80
10.50
19.00
32.00
7.78
5.10
10.30
63.00
25.80
31.00
40.00
50.00
43.66
1.34
0.47
102.8
116
< 0.001
Ratio
0.49
1.18
1.97
4.65
1
1
1
2
2
2
2
0
1
0
1
2
0
0
0
2
0
1
1
1
0
0
0
0
1
06
45
51
24
59
72
99
85
01
91
36
29
45
24
43
25
87
04
34
68
96
18
42
70
40
Do not distribute, quote or cite
B-90
Draft Document
-------
Creek chub (Semotilus atromaculatus)
Study
GEI2013
GEI2013
GEI2013
GEI2013
GEI2013
GEI2013
GEI2013
GEI2013
GEI2013
GEI2013
GEI2013
GEI2013
GEI2013
GEI2013
GEI2013
GEI2013
GEI2013
GEI2013
GEI2013
GEI2013
GEI2013
GEI2013
GEI2013
GEI2013
GEI2013
GEI2013
GEI2013
GEI2013
GEI2013
GEI2013
GEI2013
GEI2013
GEI2013
GEI2013
GEI2013
GEI2013
GEI2013
GEI2013
GEI2013
GEI2013
GEI2013
GEI2013
Site
SW4-1
SW4-1
SW4-1
SW4-1
SW4-1
SW4-1
LG1
LG1
LG1
LG1
LG1
LG1
LG1
LG1
LG1
LG1
LG1
LG1
LG1
LG1
LG1
LG1
LG1
LG1
LG1
LG1
LG1
LG1
LG1
LG1
LG1
CC1
CC1
CC1
CC1
CC1
CC1
CC1
CC1
CC1
CC1
CC1
CP
invert ^fisl
3.33
3.33
3.33
3.33
3.33
3.33
3.37
3.37
3.37
3.37
3.37
3.39
3.39
3.39
3.39
3.39
3.39
3.39
3.39
3.39
3.39
3.56
3.56
3.56
3.56
3.56
3.56
3.56
3.56
3.56
3.56
3.76
3.76
3.76
3.76
3.76
4.69
4.69
4.69
4.69
4.69
4.69
, Ratio
4.96
5.52
6.11
6.31
6.53
6.67
3.41
3.58
3.75
3.78
4.10
3.23
3.72
3.74
3.78
3.89
4.03
4.12
5.11
5.21
5.34
3.28
3.37
3.82
3.86
4.02
4.16
4.49
4.53
4.63
4.77
5.43
5.57
6.51
6.71
7.12
3.99
4.06
4.08
4.25
4.44
4.48
1.49
1.66
1.84
1.90
1.96
2.01
1.01
1.06
1.11
1.12
1.22
0.95
1.10
1.10
1.12
1.15
1.19
1.22
1.51
1.54
1.58
0.92
0.95
1.07
1.09
1.13
1.17
1.26
1.27
1.30
1.34
1.44
1.48
1.73
1.78
1.89
0.85
0.87
0.87
0.91
0.95
0.96
Do not distribute, quote or cite
B-91
Draft Document
-------
Creek chub
Study
GEI2013
GEI2013
GEI2013
GEI2013
GEI2013
GEI2013
GEI2013
GEI2013
GEI2013
GEI2013
GEI2013
GEI2013
GEI2013
GEI2013
8 -|
6 -
CM 4 -
2 -
0 -
(
(Semotilus atromaculatus)
Site Cjnvert Cflsh
CC1 4.69
CC1 4.69
CC1 4.69
CC1 4.69
CC1 5.86
CC1 5.86
CC1 5.86
CC1 5.86
CC1 5.86
CC1 5.86
CC1 5.86
CC1 5.86
CC1 5.86
CC1 5.86
o
a §«--'^ y Median ratio:
• R2:
o F:
g df:
III! p.
) 2 4 6 8
*- Invert,
Ratio
4.50
4.72
5.24
5.44
4.98
5.39
5.77
6.39
6.43
6.50
6.57
7.42
7.42
7.47
1.12
0.30
25.11
58
< 0.001
0.96
1.01
1.12
1.16
0.85
0.92
0.99
1.09
1.10
1.11
1.12
1.27
1.27
1.28
Cutthroat trout (Oncorhynchus clarkii)
Study
Site Cjnvert Cflsh
Hamilton and Buhl 2004 ShpC 1.90
McDonald and Strosher 1998 ER745 2.74
Hamilton and Buhl 2005 SC 4.10
McDonald and Strosher 1998 ER747 4.29
Hamilton and Buhl 2005 UAC 5.00
Hamilton and Buhl 2004 ACM 6.70
Hamilton and Buhl 2005 DC 8.70
McDonald and Strosher 1998 ER746 10.70
Hamilton and Buhl 2005 BGS 10.80
Hamilton and Buhl 2004 DVC 12.80
Hamilton and Buhl 2004 UEMC 26.90
Ratio
1.80
5.40
3.50
6.57
6.60
6.30
11.00
12.71
12.20
10.20
27.00
0.95
1.97
0.85
1.53
1.32
0.94
1.26
1.19
1.13
0.80
1.00
Do not distribute, quote or cite
B-92
Draft Document
-------
Cutthroat trout
Study
(Oncorhynchus clarkii)
Site
Hamilton and Buhl 2004 LEMC
60 -I
50 -
40 -
Cflsfc 30 -
20 -
10 - '
n ^
0
^
^^
o ^^^
^^^
3*5
i i i
20 40 60
P P
^ invert ^fish
75.20
X>
Median ratio:
R2:
F:
df:
1 P:
80
Ratio
52.30
1.07
0.97
325.2
10
< 0.001
0.70
Fathead minnow (Pimephales promelas)
Study
Butler etal. 1997
Butler etal. 1997
Butler etal. 1995
Butler etal. 1995
Butler etal. 1995
Butler etal. 1995
Butler etal. 1995
Butler etal. 1995
Butler etal. 1995
Butler etal. 1995
Butler etal. 1995
Butler etal. 1995
Butler etal. 1995
Butler etal. 1993
Butler etal. 1993
Butler etal. 1995
Butler etal. 1995
Butler etal. 1995
Butler etal. 1995
Butler etal. 1995
Butler etal. 1995
Butler etal. 1997
Butler etal. 1997
Butler etal. 1997
Site
MNP1
MNP1
AK
AK
AK
HD1
HD1
HD1
DD
DD
DD
HD2
HD2
Dl
Dl
ME2
SD
SD
SD
ME4
ME4
TRH
TRH
TRH
Cp
invert ^fish
0.70
0.70
0.78
0.78
0.78
0.83
0.83
0.83
0.86
0.86
0.86
0.98
0.98
1.20
1.20
1.25
1.40
1.40
1.40
1.55
1.55
1.60
1.60
1.60
Ratio
1.70
1.80
2.60
2.80
2.90
2.50
2.60
3.90
3.40
3.60
3.90
1.50
1.60
3.70
3.80
4.80
3.00
4.00
4.90
1.40
5.90
2.20
3.00
4.20
2.43
2.57
3.35
3.61
3.74
3.03
3.15
4.73
3.95
4.19
4.53
1.53
1.63
3.08
3.17
3.84
2.14
2.86
3.50
0.90
3.81
1.38
1.88
2.63
Do not distribute, quote or cite
B-93
Draft Document
-------
Fathead minnow (Pimephales promelas)
Study
Butler etal. 1997
Butler etal. 1995
Butler etal. 1995
Birkner 1978
Birkner 1978
Butler etal. 1997
Butler etal. 1997
Butler etal. 1997
Grassoetal. 1995
Grassoetal. 1995
Grassoetal. 1995
Grassoetal. 1995
Grassoetal. 1995
Grassoetal. 1995
Butler etal. 1993
Butler etal. 1995
Butler etal. 1995
Butler etal. 1995
Butler etal. 1994
GEI2013
GEI2013
GEI2013
GEI2013
GEI2013
GEI2013
GEI2013
GEI2013
GEI2013
GEI2013
Butler etal. 1993
Lambing etal. 1994
Butler etal. 1993
GEI2013
GEI2013
GEI2013
GEI2013
GEI2013
GEI2013
GEI2013
GEI2013
GEI2013
GEI2013
Site
TRH
YJ2
YJ2
1
4
TR25
TR25
TR25
10
10
10
17
17
17
Ul
ME3
ME3
ME3
AD
SWA1
SWA1
SWA1
SWA1
SWA1
SWA1
SWA1
SWA1
SWA1
SWA1
WSB2
S48
SP2
SW4-1
SW4-1
SW4-1
SW4-1
SW4-1
SW4-1
SW4-1
SW4-1
SW4-1
SW4-1
c c
^^ invert ^fis
1.60
1.65
1.65
1.75
1.80
1.80
1.80
1.80
1.85
1.85
1.85
1.91
1.91
1.91
2.45
2.55
2.55
2.55
2.70
2.81
2.81
2.81
2.81
2.81
2.81
2.81
2.81
2.81
2.81
3.00
3.05
3.15
3.33
3.33
3.33
3.33
3.33
3.33
3.33
3.33
3.33
3.33
h Ratio
4.30
4.00
11.00
2.10
2.10
4.00
5.20
6.00
2.74
2.79
2.90
6.59
6.60
7.30
6.40
4.30
4.40
5.30
9.60
3.89
3.98
4.04
4.33
4.48
4.53
4.81
5.00
5.24
5.76
8.10
2.50
8.20
4.85
5.25
5.39
5.88
5.89
6.07
6.11
6.61
6.67
6.87
2.69
2.42
6.67
1.20
1.17
2.22
2.89
3.33
1.48
1.51
1.57
3.45
3.46
3.82
2.61
1.69
1.73
2.08
3.56
1.39
1.42
1.44
1.54
1.60
1.61
1.71
1.78
1.87
2.05
2.70
0.82
2.60
1.46
1.58
1.62
1.77
1.77
1.83
1.84
1.99
2.01
2.07
Do not distribute, quote or cite
B-94
Draft Document
-------
Fathead minnow (Pimephales promelas)
Study
GEI2013
GEI2013
GEI2013
GEI2013
GEI2013
GEI2013
Butler et al.
Butler et al.
Butler et al.
Butler et al.
Butler et al.
GEI2013
GEI2013
GEI2013
GEI2013
GEI2013
GEI2013
GEI2013
GEI2013
GEI2013
GEI2013
Butler et al.
Butler et al.
Butler et al.
GEI2013
GEI2013
GEI2013
GEI2013
GEI2013
GEI2013
GEI2013
GEI2013
GEI2013
GEI2013
Butler et al.
Butler et al.
Butler et al.
Butler et al.
Butler et al.
GEI2013
GEI2013
GEI2013
1993
1995
1997
1997
1997
1993
1993
1994
1993
1993
1994
1993
1993
Site
LG1
LG1
LG1
LG1
LG1
LG1
SP2
ME1
MUD2
MUD2
MUD2
LG1
LG1
LG1
LG1
LG1
LG1
LG1
LG1
LG1
LG1
WSB2
WSB2
CF1
SWA1
SWA1
SWA1
SWA1
SWA1
SWA1
SWA1
SWA1
SWA1
SWA1
SB2
R2
PSW1
SB2
SB2
CC1
CC1
CC1
c
^^ invert
3
3
3
3
3
3
3
3
3
3
3
3
3
3
3
3
3
3
3
3
3
3
3
3
3
3
3
3
3
3
3
3
3
3
3
3
3
3
3
3
3
3
Cfish
.39
.39
.39
.39
.39
.39
.40
.40
.45
.45
.45
.56
.56
.56
.56
.56
.56
.56
.56
.56
.56
.60
.60
.60
.64
.64
.64
.64
.64
.64
.64
.64
.64
.64
.65
.70
.70
.75
.75
.76
.76
.76
3
3
4
4
5
5
6
5
6
7
12
3
3
3
4
4
4
4
5
5
5
4
10
7
3
3
4
4
4
4
4
4
5
5
9
6
22
5
8
3
5
7
Ratio
.60
.89
.27
.45
.18
.51
.00
.60
.50
.70
.00
.26
.35
.72
.09
.20
.81
.86
.05
.47
.56
.20
.00
.90
.62
.72
.07
.43
.52
.66
.68
.76
.45
.71
.90
.60
.00
.70
.60
.79
.23
.36
1.06
1.15
1.26
1.31
1.53
1.63
1.76
1.65
1.88
2.23
3.48
0.92
0.94
1.05
1.15
1.18
1.35
1.37
1.42
1.54
1.56
1.17
2.78
2.19
1.00
1.02
1.12
1.22
1.24
1.28
1.29
1.31
1.50
1.57
2.71
1.78
5.95
1.52
2.29
1.01
1.39
1.96
Do not distribute, quote or cite
B-95
Draft Document
-------
Fathead minnow (Pimephales promelas)
Study
GEI2013
GEI2013
Butler et al.
Butler et al.
GEI2013
GEI2013
GEI2013
GEI2013
GEI2013
GEI2013
GEI2013
GEI2013
Butler et al.
Butler et al.
Butler et al.
Butler et al.
Butler et al.
GEI2013
GEI2013
GEI2013
GEI2013
GEI2013
GEI2013
Butler et al.
GEI2013
GEI2013
GEI2013
GEI2013
GEI2013
Butler et al.
Butler et al.
Butler et al.
Lemly 1985
Lambing et
Lambing et
GEI2013
GEI2013
GEI2013
GEI2013
GEI2013
GEI2013
GEI2013
1993
1994
1993
1993
1993
1993
1997
1993
1993
1991
1994
al. 1994
al. 1994
Site
CC1
CC1
R2
LSW1
SW88
SW88
SW88
SW88
SW88
SW88
SW88
SW88
Rl
Rl
ST2
ST2
MNP2
SW9
SW9
SW9
SW9
SW9
SW9
ST2
CC1
CC1
CC1
CC1
CC1
WSB2
10
TGC
Badin Lake
S39
S39
CC1
CC1
CC1
CC1
CC1
CC1
CC1
c c
^^ invert ^fis
3.76
3.76
3.90
3.90
3.96
3.96
3.96
3.96
3.96
3.96
3.96
3.96
4.00
4.00
4.10
4.10
4.40
4.45
4.45
4.45
4.45
4.45
4.45
4.50
4.69
4.69
4.69
4.69
4.69
4.75
4.80
4.90
5.18
5.85
5.85
5.86
5.86
5.86
5.86
5.86
5.86
5.86
lh Ratio
8.69
9.07
6.60
73.00
4.73
4.96
5.13
5.55
5.56
5.86
6.07
6.32
11.00
11.00
7.60
16.00
11.00
5.57
5.93
6.14
6.20
6.56
7.57
12.80
5.92
6.49
7.14
7.59
7.68
17.10
8.10
11.00
2.54
7.90
21.00
6.68
7.73
7.88
8.45
9.21
9.70
11.69
2.31
2.41
1.69
18.72
1.20
1.25
1.30
1.40
1.41
1.48
1.53
1.60
2.75
2.75
1.85
3.90
2.50
1.25
1.33
1.38
1.39
1.47
1.70
2.84
1.26
1.39
1.52
1.62
1.64
3.60
1.69
2.24
0.49
1.35
3.59
1.14
1.32
1.35
1.44
1.57
1.66
2.00
Do not distribute, quote or cite
B-96
Draft Document
-------
Fathead minnow (Pimephales promelas)
Study
Butler etal. 1991
Lambing etal. 1994
GEI2013
GEI2013
GEI2013
GEI2013
GEI2013
GEI2013
GEI2013
GEI2013
GEI2013
GEI2013
GEI2013
GEI2013
GEI2013
GEI2013
GEI2013
GEI2013
GEI2013
GEI2013
GEI2013
Butler etal. 1995
Butler etal. 1995
Butler etal. 1995
Lemly 1985
GEI2013
GEI2013
GEI2013
GEI2013
GEI2013
GEI2013
GEI2013
GEI2013
GEI2013
GEI2013
GEI2013
GEI2013
GEI2013
GEI2013
GEI2013
GEI2013
GEI2013
Site
3
S46
SW1
SW1
SW1
SW1
SW1
SW1
SW1
SW1
SW1
SW2-1
SW2-1
SW2-1
SW2-1
SW2-1
SW2-1
SW2-1
SW2-1
SW2-1
SW2-1
we
we
we
High Rock Lake
SWB
SWB
SWB
SWB
SWB
SWB
SWB
SWB
SWB
SWB
SWB
SWB
SWB
SWB
SWB
SWB
SWB
c c
^^ invert ^fis
6.20
6.20
6.54
6.54
6.54
6.54
6.54
6.54
6.54
6.54
6.54
6.60
6.60
6.60
6.60
6.60
6.60
6.60
6.60
6.60
6.60
6.75
6.75
6.75
6.75
7.06
7.06
7.06
7.06
7.06
7.06
7.06
7.06
7.06
7.44
7.44
7.44
7.44
7.44
7.44
7.44
7.44
;h Ratio
9.50
5.10
7.01
7.86
7.98
8.23
8.50
9.48
9.95
10.09
10.19
12.51
12.83
14.80
16.70
17.21
18.27
20.13
20.66
26.75
30.48
18.40
22.90
26.40
3.21
7.38
8.49
8.61
8.72
9.02
9.11
9.30
9.53
9.80
9.36
9.46
9.78
9.87
10.66
10.97
11.22
12.25
1.53
0.82
1.07
1.20
1.22
1.26
1.30
1.45
1.52
1.54
1.56
1.90
1.95
2.24
2.53
2.61
2.77
3.05
3.13
4.06
4.62
2.73
3.39
3.91
0.48
1.05
1.20
1.22
1.24
1.28
1.29
1.32
1.35
1.39
1.26
1.27
1.32
1.33
1.43
1.48
1.51
1.65
Do not distribute, quote or cite
B-97
Draft Document
-------
Fathead minnow (Pimephales promelas)
Study
GEI2013
GEI2013
Butler etal. 1994
GEI2013
GEI2013
GEI2013
GEI2013
GEI2013
GEI2013
GEI2013
GEI2013
GEI2013
GEI2013
Butler etal. 1994
GEI2013
GEI2013
GEI2013
GEI2013
GEI2013
GEI2013
GEI2013
Butler etal. 1997
GEI2013
GEI2013
GEI2013
GEI2013
GEI2013
GEI2013
GEI2013
GEI2013
Butler etal. 1997
Butler etal. 1997
Birkner 1978
Lambing etal. 1994
Lambing etal. 1994
Lambing etal. 1994
Birkner 1978
Butler etal. 1994
Butler etal. 1994
Birkner 1978
Lemly 1985
Butler etal. 1994
Site
SWB
SWB
CRC
SW1
SW1
SW1
SW1
SW1
SW1
SW1
SW1
SW1
SW1
IW
SW11
SW11
SW11
SW11
SW11
SW11
SW11
MN5
SW2-1
SW2-1
SW2-1
SW2-1
SW2-1
SW2-1
SW2-1
SW2-1
WCP
WCP
22
S34
Sll
Sll
23
GUN2
MKP
27
Belews Lake
OMD
c c
^^ invert ^fis
7.44
7.44
7.50
7.82
7.82
7.82
7.82
7.82
7.82
7.82
7.82
7.82
7.82
8.35
8.41
8.41
8.41
8.41
8.41
8.41
8.41
8.60
9.14
9.14
9.14
9.14
9.14
9.14
9.14
9.14
9.70
9.70
11.30
14.00
14.50
14.50
15.50
28.00
32.00
34.60
45.53
73.00
;h Ratio
12.43
12.46
20.40
8.45
8.88
9.11
9.15
9.41
9.82
11.07
11.15
11.23
13.76
10.00
4.68
5.29
5.34
5.38
5.38
5.70
7.05
7.30
13.31
15.63
15.77
16.79
17.00
18.21
19.39
22.50
10.00
15.00
11.00
25.00
11.00
33.00
34.50
7.50
51.00
79.00
23.03
13.00
1.67
1.68
2.72
1.08
1.14
1.16
1.17
1.20
1.26
1.42
1.43
1.44
1.76
1.20
0.56
0.63
0.63
0.64
0.64
0.68
0.84
0.85
1.46
1.71
1.73
1.84
1.86
1.99
2.12
2.46
1.03
1.55
0.97
1.79
0.76
2.28
2.23
0.27
1.59
2.28
0.51
0.18
Do not distribute, quote or cite
B-98
Draft Document
-------
Fathead minnow
Study
100 -|
80 - Q
60 -
Cfish 40 -
B
20 -91
A -^^K
U i—
0
(Pimephales promelas)
Site
o
° ^^^^
\f ^^-^^
Ixr"'""""^ o
P" O
20 40 60 80
c c
^invert ^fish
Median ratio:
R2:
F:
df:
P:
Ratio
1.57
0.22
64.4
232
< 0.001
Flannelmouth sucker (Catostomus latipinnis)
Study
Butler etal. 1995
Butler etal. 1995
Butler etal. 1995
Butler etal. 1995
Butler etal. 1995
Butler etal. 1995
Butler etal. 1995
Butler etal. 1995
Butler etal. 1995
Butler etal. 1993
Butler etal. 1993
Butler etal. 1995
Butler etal. 1995
Butler etal. 1995
Butler etal. 1995
Butler etal. 1993
Butler etal. 1994
Butler etal. 1994
Butler etal. 1994
Butler etal. 1994
Butler etal. 1994
Butler etal. 1994
Butler etal. 1994
Butler etal. 1994
Butler etal. 1994
Site
AK
AK
AK
AK
HD1
HD2
HD2
HD2
HD2
LP3
LP3
ME2
ME2
ME2
ME2
PI
COL1
COL1
COL1
COL1
COL1
COL1
COL1
COL1
COL1
P P
^^ invert ^fish
0.78
0.78
0.78
0.78
0.83
0.98
0.98
0.98
0.98
1.12
1.12
1.25
1.25
1.25
1.25
1.50
1.50
1.50
1.50
1.50
1.50
1.50
1.50
1.50
1.50
Ratio
0.82
0.90
1.10
1.10
2.90
0.49
0.54
0.62
0.96
0.92
1.40
1.40
1.60
2.00
2.20
2.40
0.50
0.60
0.63
0.92
.00
.60
.70
.80
.90
1.06
1.16
1.42
1.42
3.52
0.50
0.55
0.63
0.98
0.83
1.26
1.12
1.28
1.60
1.76
1.60
0.33
0.40
0.42
0.61
0.67
1.07
1.13
1.20
1.27
Do not distribute, quote or cite
B-99
Draft Document
-------
Flannelmouth sucker (Catostomus latipinnis)
Study
Butler
Butler
Butler
Butler
Butler
Butler
Butler
Butler
Butler
Butler
Butler
Butler
Butler
Butler
Butler
Butler
Butler
Butler
Butler
Butler
Butler
Butler
Butler
Butler
Butler
Butler
Butler
Butler
Butler
Butler
Butler
Butler
Butler
Butler
Butler
Butler
Butler
Butler
Butler
Butler
Butler
Butler
et
et
et
et
et
et
et
et
et
et
et
et
et
et
et
et
et
et
et
et
et
et
et
et
et
et
et
et
et
et
et
et
et
et
et
et
et
et
et
et
et
et
al.
al.
al.
al.
al.
al.
al.
al.
al.
al.
al.
al.
al.
al.
al.
al.
al.
al.
al.
al.
al.
al.
al.
al.
al.
al.
al.
al.
al.
al.
al.
al.
al.
al.
al.
al.
al.
al.
al.
al.
al.
al.
1994
1995
1995
1995
1995
1995
1994
1995
1995
1995
1995
1997
1997
1997
1993
1997
1997
1995
1995
1995
1995
1995
1995
1995
1995
1995
1995
1995
1997
1997
1995
1995
1995
1997
1997
1993
1993
1994
1994
1991
1991
1991
Site
COL1
ME4
ME4
ME4
ME4
ME4
RB3
MP
MP
YJ2
YJ2
MNQ
MNQ
MNQ
PI
MUD
MUD
SJ1
SJ1
SJ1
SJ1
SJ1
SJ1
ME3
ME3
ME3
ME3
ME3
MN4
MN4
MN1
MN1
MN1
MN3
MN3
LP4
LP4
PSW1
LSW1
4
9
9
p
^ invert
1
1
1
1
1
1
1
1
1
1
1
1
1
1
1
2.
2.
2.
2.
2.
2.
2.
2.
2.
2.
2.
2.
2.
2.
2.
2.
2.
2.
2.
2.
3
3
3
3
3
4
4
Cfish
.50
.55
.55
.55
.55
.55
.60
.60
.60
.65
.65
.80
.80
.80
.95
.30
.30
.50
.50
.50
.50
.50
.50
.55
.55
.55
.55
.55
.65
.65
.70
.70
.70
.70
.70
.20
.20
.70
.90
.90
.10
.10
1
1
1
1
2
3
29
1
1
1
2
2
3
3
1
2
4
0
1
1
2
3
4
1
1
2
2
3
5
9
1
4
6
2
2
2
2
9
6
1
1
6
Ratio
.90
.30
.50
.90
.40
.00
.00
.20
.40
.60
.40
.10
.20
.50
.50
.70
.10
.61
.10
.50
.20
.19
.20
.70
.70
.10
.40
.60
.10
.60
.70
.80
.50
.30
.60
.40
.60
.40
.70
.70
.50
.00
1
0
0
1
1
1
18
0
0
0
1
1
1
1
0
1
1
0
0
0
0
1
1
0
0
0
0
1
1
3
0
1
2
0
0
0
0
2
1
0
0
1
.27
.84
.97
.23
.55
.94
.13
.75
.88
.97
.45
.17
.78
.94
.77
.17
.78
.24
.44
.60
.88
.27
.68
.67
.67
.82
.94
.41
.92
.62
.63
.78
.41
.85
.96
.75
.81
.54
.72
.44
.37
.46
Do not distribute, quote or cite
B-100
Draft Document
-------
Flannelmouth sucker (Catostomus latipinnis)
Study
Butler
Butler
Butler
Butler
Butler
Butler
Butler
Butler
£
etal. 1991
etal. 1994
etal. 1994
etal. 1997
etal. 1997
etal. 1994
etal. 1994
etal. 1991
35 -
30 -
25 -
20 -
sh 15 -
10 -
5 -
0 -
O
O n
coo °-
Jp
W£=-— |
0 10
c
Site
10
BSW1
CRC
MN5
NW2
LZA1
RBI
7
o
-^^"^
0
1 '
20 30
>l
"invert.
c
*^ invert
4
5
7
8
11
19
21
29
£
80
00
50
60
40
00
00
80
Median ratio:
1
40
R2:
F:
df:
P:
Ratio
2.50
9.60
12.00
8.40
11.00
17.00
4.60
22.00
1.06
0.36
41.6
73
< 0.001
0.52
1.92
1.60
0.98
0.96
0.89
0.22
0.74
Do not distribute, quote or cite
B-101
Draft Document
-------
Gizzard shad (Dorosoma cepedianum)
Study
Mueller et al.
Mueller et al.
Mueller et al.
20 -
15 -
Cfish 10 -
5 -
0 -
1991
1991
1991
i i
024
Site
R2
Rl
Rl
a
"S
1
6
c
^-'invert.
C C
^invert ^fish
6.40
8.70
8.70
s. Median ratio:
^x
0 R2:
F:
df:
p.
8 10 Not used because P >
slope.
14.30
7.50
11.00
1.26
0.74
2.78
1
0.39
Ratio
2.23
0.86
1.26
0.05 and negative
Goldeye (Hiodon alosoides)
Study
Roddy etal. 1991
Roddy etal. 1991
Roddy etal. 1991
Roddy etal. 1991
Roddy etal. 1991
Roddy etal. 1991
Roddy etal. 1991
Roddy etal. 1991
Roddy etal. 1991
Roddy etal. 1991
Site
18
18
18
18
18
18
18
18
18
18
Cp
invert ^fish
3.10
3.10
3.10
3.10
3.10
3.10
3.10
3.10
3.10
3.10
2.00
2.10
2.20
2.30
2.40
2.70
2.90
3.40
3.60
4.70
Ratio
0.65
0.68
0.71
0.74
0.77
0.87
0.94
1.10
1.16
1.52
Do not distribute, quote or cite
B-102
Draft Document
-------
Goldeye (Hiodon alosoides)
Study
5 ,
4 -
3 -
fish 2 _
1 -
1
0
Site
o
8
Q
a
i
i i i
1 2 3
c
^-'invert.
C C
^^ invert ^fish
Median ratio:
R2:
F:
df:
4 P:
Ratio
0.82
0.0
0.0
8
1.0
Green sunfish (Lepomis cyanellus)
Study
Butler etal. 1995
Butler etal. 1995
Butler etal. 1995
Butler etal. 1997
Butler etal. 1997
Butler etal. 1995
GEI2013
GEI2013
GEI2013
GEI2013
GEI2013
Roddy etal. 1991
Roddy etal. 1991
Roddy etal. 1991
Roddy etal. 1991
Roddy etal. 1991
GEI2013
GEI2013
GEI2013
Butler etal. 1997
Butler etal. 1997
GEI2013
GEI2013
GEI2013
GEI2013
Site
HD1
HD1
MP
TRH
TR25
ME3
SWA1
SWA1
SWA1
SWA1
SWA1
18
18
18
18
18
LG1
LG1
LG1
MUD2
MUD2
SW88
SW88
SW9
SW9
Cp
invert ^fish
0.83
0.83
1.60
1.60
1.80
2.55
2.81
2.81
2.81
2.81
2.81
3.10
3.10
3.10
3.10
3.10
3.39
3.39
3.39
3.45
3.45
3.96
3.96
4.45
4.45
Ratio
1.30
1.30
1.90
3.30
4.40
5.00
2.96
3.21
3.24
3.69
3.88
2.80
3.80
4.00
5.20
5.70
4.11
4.33
5.71
7.00
7.60
7.14
7.41
4.38
5.06
1.58
1.58
1.19
2.06
2.44
1.96
1.06
1.14
1.16
1.32
1.38
0.90
1.23
1.29
1.68
1.84
1.21
1.28
1.68
2.03
2.20
1.81
1.87
0.98
1.14
Do not distribute, quote or cite
B-103
Draft Document
-------
Green sunfish (Lepomis cyanellus)
GEI2013
GEI2013
GEI2013
SW9
SW9
SW9
Butler etal. 1991 10
Lemly 1985
Badin Lake
Butler etal. 1991 3
Lemly 1985
GEI2013
High Rock Lake
SWB
Butler etal. 1997 CHI
GEI2013
GEI2013
GEI2013
GEI2013
GEI2013
SW11
SW11
SW11
SW11
SW11
Butler etal. 1994 LZA1
Butler etal. 1991 7
Butler etal. 1991 7
Lemly 1985
40 -
30 -
C 20 -
10 -
A .
Belews Lake
o
4
4
4
4
5
6
6
7
7
8
8
8
8
8
19
29
29
45
45
45
45
80
18
20
75
44
50
41
41
41
41
41
00
80
80
53
Median ratio:
o ^^-""""""'^
.^^ 0
^^*^ °
gftjb^'^
3Ki3*
3*
R2:
F:
df:
P:
5.53
5.80
7.29
7.90
3.43
6.40
3.30
11.94
9.50
4.54
4.84
5.34
7.00
7.13
37.00
15.20
25.10
21.96
1.27
0.59
57.9
41
< 0.001
1
1
1
1
0
1
0
1
1
0
0
0
0
0
1
0
0
0
24
30
64
65
66
03
49
61
27
54
58
63
83
85
95
51
84
48
10 20 30 40 50
p
'-invert.
Do not distribute, quote or cite
B-104
Draft Document
-------
Iowa darter (Etheostoma exile)
Study
Birkner 1978
Birkner 1978
Birkner 1978
Birkner 1978
50 -
40 -
30 -
(^ .
fish 2Q _
10 -
0 -
Site
7
20
22
23
i i i i
C C
^invert ^fish
3.75
11.20
11.30
15.50
Median ratio:
R2:
F:
df:
P:
Not used because P >
Ratio
2.10
36.30
23.00
41.90
2.37
0.90
17.3
2
0.055
0.05
0.56
3.24
2.04
2.70
0 5 10 15 20
Largemouth
Study
C
*- invert.
bass (Micropterm salmoides)
Site
Saikietal. 1993 ET6
Saikietal. 1993 ET6
Saikietal. 1993 ET7
Saikietal. 1993 ET7
Saikietal. 1993 SJR1
Saikietal. 1993 SJR1
Rinella and Schuler 1 992 Malheur Lake
Saikietal. 1993 SJR3
Saikietal. 1993 SJR3
Butler etal. 1995 MP
GEI2013
SWA1
Garcia-Hernandez et al. 2000 Cienga Wetland
Saikietal. 1993 SJR2
Saikietal. 1993 SJR2
GEI2013
GEI2013
GEI2013
GEI2013
GEI2013
SW4-1
SW4-1
SW4-1
SW4-1
SW4-1
C C
^^ invert ^fish
0.85
0.85
0.86
0.86
0.95
0.95
1.20
1.50
1.50
1.60
2.81
3.00
3.30
3.30
3.33
3.33
3.33
3.33
3.33
Ratio
1.00
1.40
0.86
1.00
0.80
1.80
0.92
1.70
1.80
1.40
3.17
5.10
2.20
2.40
5.53
5.65
5.72
5.80
6.34
1.18
1.66
1.00
1.16
0.85
1.90
0.77
1.13
1.20
0.88
1.13
1.70
0.67
0.73
1.66
1.70
1.72
1.74
1.91
Do not distribute, quote or cite
B-105
Draft Document
-------
Largemouth bass (Micropterm salmoides)
Study
GEI2013
GEI2013
GEI2013
GEI2013
GEI2013
GEI2013
GEI2013
GEI2013
Saikietal. 1993
Saikietal. 1993
GEI2013
GEI2013
GEI2013
GEI2013
GEI2013
GEI2013
Saikietal. 1993
Saikietal. 1993
GEI2013
GEI2013
GEI2013
GEI2013
GEI2013
GEI2013
Crutchfield 2000
Crutchfield 2000
Crutchfield 2000
Crutchfield 2000
Crutchfield 2000
Crutchfield 2000
Crutchfield 2000
Crutchfield 2000
Crutchfield 2000
Crutchfield 2000
Crutchfield 2000
Crutchfield 2000
Crutchfield 2000
Crutchfield 2000
Crutchfield 2000
Crutchfield 2000
Crutchfield 2000
Crutchfield 2000
Site
SW4-1
LG1
SW88
SW88
SW88
SW88
SW88
SW88
GT4
GT4
SW9
SW9
SW9
SW9
SW9
SW9
GT5
GT5
SW11
SW11
SW11
SW11
SW11
SW11
transect 3
transect 3
transect 3
transect 3
transect 3
transect 3
transect 3
transect 3
transect 3
transect 3
transect 3
transect 3
transect 4
transect 4
transect 4
transect 4
transect 4
transect 4
P P
^ invert ^fis
3.33
3.39
3.96
3.96
3.96
3.96
3.96
3.96
4.05
4.05
4.45
4.45
4.45
4.45
4.45
4.45
4.90
4.90
8.41
8.41
8.41
8.41
8.41
8.41
8.60
8.60
8.60
8.60
9.25
9.25
9.25
9.25
11.40
11.40
11.95
11.95
15.70
15.70
15.70
15.70
16.45
16.45
;h Ratio
7.14
4.29
4.87
5.73
5.77
5.93
6.62
6.84
4.00
4.70
5.78
5.79
6.19
6.87
7.27
7.36
6.80
6.90
5.02
5.19
5.77
6.26
6.48
7.22
8.92
9.50
10.32
11.40
10.70
10.96
14.00
14.20
23.70
26.12
19.62
22.30
13.50
14.78
29.69
34.90
10.70
11.20
2.15
1.27
1.23
1.45
1.46
1.50
1.67
1.73
0.99
1.16
1.30
1.30
1.39
1.54
1.63
1.65
1.39
1.41
0.60
0.62
0.69
0.74
0.77
0.86
1.04
1.10
1.20
1.33
1.16
1.18
1.51
1.54
2.08
2.29
1.64
1.87
0.86
0.94
1.89
2.22
0.65
0.68
Do not distribute, quote or cite
B-106
Draft Document
-------
Largemouth
Study
bass (Micropterm salmoides)
Site
Crutchfield 2000 transect 4
Crutchfield 2000 transect 4
Crutchfield 2000 transect 4
Crutchfield 2000 transect 4
Crutchfield 2000 transect 4
Crutchfield 2000 transect 4
40 -I
30 -
Cfish 20 "
10 -
0 -
o
o 8
o °
o ^^**®
o^***^% 8
e&**^£ °
—^g^"^ *
^^u ^*
11111
P P
^ invert ^fish
18.25
18.25
20.90
20.90
20.90
20.90
Median ratio:
R2:
F:
df:
P:
Ratio
14.78
17.20
20.20
24.34
28.60
30.83
1.27
0.72
164.3
65
< 0.001
0.81
0.94
0.97
1.16
1.37
1.48
0 5 10 15 20 25
Longnose dace (Rhinichthys cataractae)
Study
Site
Lambing etal. 1994 S3 3
Mueller etal. 1991 Al
GEI2013 LG1
GEI2013
GEI2013
GEI2013
GEI2013
GEI2013
GEI2013
GEI2013
GEI2013
GEI2013
GEI2013
GEI2013
Mueller et al.
Hamilton and
Hamilton and
LG1
LG1
LG1
LG1
LG1
LG1
LG1
LG1
LG1
LG1
LG1
1991 Tl
Buhl 2005 CC
Buhl 2005 BGS
Cp
invert ^fish
2.40
2.70
3.37
3.37
3.37
3.37
3.37
3.39
3.39
3.56
3.56
3.56
3.56
3.56
5.40
6.70
10.80
Ratio
5.30
2.10
5.05
5.57
6.57
6.75
10.08
10.69
12.77
8.95
9.63
11.41
11.94
12.04
16.90
13.40
10.90
2.21
0.78
1.50
1.65
1.95
2.00
2.99
3.15
3.77
2.52
2.71
3.21
3.36
3.39
3.13
2.00
1.01
Do not distribute, quote or cite
B-107
Draft Document
-------
Longnose dace (Rhinichthys cataractae)
Study
20 -I
15 -
QI
cfisn 10 - jt
5 - O |
O
n
0
Site
o
Q^^*
^^^ °
5 10
^-Invert.
c c
*^ invert ^fish
Median ratio:
R2:
F:
df:
P:
15 Not used because P >
Ratio
2.52
0.17
3.16
15
0.07
0.05
Longnose sucker (Catostomus catostomus)
Study
Minnow 2007
Butler etal. 1994
Butler etal. 1994
Butler etal. 1994
Butler etal. 1994
Butler etal. 1994
Butler etal. 1994
Butler etal. 1994
Butler etal. 1994
Butler etal. 1994
Butler etal. 1994
Mueller etal. 1991
Minnow 2007
10 -
8 -
6 -
tKh 4 " £ ^0"
") - 8
o
T i
Site
FL17
NFK2
NFK2
NFK2
NFK2
NFK2
NFK2
NFK2
NFK2
NFK2
NFK2
Tl
FL17
^®
^^^
^^
i i i
Cp
invert ^fish
3.03
3.10
3.10
3.10
3.10
3.10
3.10
3.10
3.10
3.10
3.10
5.40
21.22
Median ratio:
R2:
F:
df:
1 P:
Ratio
1.40
2.10
2.50
2.70
2.80
2.90
3.00
3.20
3.30
3.40
4.00
3.60
7.90
0.90
0.83
54.7
11
<0.001
0.46
0.68
0.81
0.87
0.90
0.94
0.97
1.03
1.06
1.10
1.29
0.67
0.37
10 15
1
"Invert
20
25
Do not distribute, quote or cite
B-108
Draft Document
-------
Mosquitofish (Gambusia sp.)
Study
Lemly 1985
Lemly 1985
Lemly 1985
50 -i
40 •
30 -
cflsh 2Q _
J**
10 - .^
oer
1 1
0 10
Site
Badin Lake
High Rock Lake
Belews Lake
^
/^
^^
/^
^
i i i i
20 30 40 50
C
^ in vert.
C C
^ invert ^fish
5.18
6.75
45.53
Median ratio:
R2:
F:
df:
P:
Ratio
5.44
5.75
44.15
0.97
1.00
1326
1
0.019
1.05
0.85
0.97
Mottled sculpin (Cottus bairdii)
Study
Hamilton and Buhl 2004
Butler etal. 1993
Butler etal. 1993
Butler etal. 1993
Butler etal. 1993
Butler etal. 1993
Butler etal. 1993
Butler etal. 1993
Butler etal. 1993
Hamilton and Buhl 2004
Butler etal. 1993
Butler etal. 1994
Butler etal. 1997
Butler etal. 1997
Butler etal. 1997
Lambing etal. 1994
Butler etal. 1991
Butler etal. 1997
Butler etal. 1997
Butler etal. 1994
Site
use
LP2
LP2
LP3
LP3
LP3
PI
PI
PI
ShpC
PI
NFK3
MN2
CHK
CHK
S33
12
MN1
MN1
NFK2
C C
^invert ^fish
0.50
1.00
1.00
1.12
1.12
1.12
1.50
1.50
1.50
1.90
1.95
2.00
2.20
2.40
2.40
2.40
2.80
2.90
2.90
3.10
Ratio
5.30
2.20
3.10
3.90
4.20
4.90
5.10
6.40
6.70
4.10
7.30
5.80
2.60
3.10
4.40
3.70
4.20
3.20
3.40
6.40
10.60
2.20
3.10
3.50
3.77
4.39
3.40
4.27
4.47
2.16
3.74
2.90
1.18
1.29
1.83
1.54
1.50
1.10
1.17
2.06
Do not distribute, quote or cite
B-109
Draft Document
-------
Mottled sculpin (Cottus bairdii)
Study
Site Cjnvert Cflsh
Butler etal. 1991 4 3.90
Butler etal. 1991 4 3.90
Butler etal. 1991 10 4.80
Hamilton and
Buhl 2005 UAC 5.00
Butler etal. 1991 3 6.20
Hamilton and
Hamilton and
Buhl 2004 ACM 6.70
Buhl 2005 CC 6.70
Butler etal. 1993 F2 7.50
Hamilton and
Hamilton and
Hamilton and
Hamilton and
Buhl 2004 LBR 7.70
Buhl 2005 DC 8.70
Buhl 2005 BGS 10.80
Buhl 2004 DVC 12.80
Butler etal. 1994 HCC1 21.00
14 -
12 -
10 -
R -
Cflsh
O
4 -
^
0 -
o ®
^__^ Median ratio:
O ^-2""""'"'""^
£o oP---'"""""""'""'""^ R2
QsBc — o^o ® F:
gPo df:
P:
11111
Ratio
2.60
4.40
5.00
6.20
6.50
8.30
8.20
9.90
5.20
12.00
12.30
8.80
5.60
1.38
0.27
11.62
31
< 0.001
0.67
1.13
1.04
1.24
1.05
1.24
1.22
1.32
0.68
1.38
1.14
0.69
0.27
0 5 10 15 20 25
C
^-invert.
Mountain whitefish (Prosopium williamsoni)
Study
Site Cjnvert Cflsh
Low and Mullins 1990 7 1.60
McDonald and Strosher 1998 ER745 2.74
Minnow 2007 EL12 4.01
McDonald and Strosher 1998 ER747 4.29
Minnow 2007 MI3 6.21
Minnow 2007 MI2 6.69
Minnow 2007 ELI 7.08
Minnow 2007 FO23 10.00
Ratio
1.40
4.17
6.60
4.93
9.12
10.16
9.12
10.20
0.88
1.52
1.65
1.15
1.47
1.52
1.29
1.02
Do not distribute, quote or cite
B-110
Draft Document
-------
Mountain whitefish (Prosopium williamsoni)
Study
Site
Cjnvert
Cfl
lsh
Ratio
14 -
12 -
10 -
8 -
6 -
4 -
2 •
0
Median ratio: 1.38
0
10
15
R2.
F:
df:
P:
0.83
30.3
6
<0.001
c
*^ invert.
Do not distribute, quote or cite
B-lll
Draft Document
-------
Northern pike (Esox lucius)
Study
Butler etal. 1995
Butler etal. 1995
Butler etal. 1995
Butler etal. 1995
Butler etal. 1995
Butler etal. 1995
Butler etal. 1995
Butler etal. 1995
j.5
3.0
-j S
2.0
CF"* 1.5 Q
•^
1.0 Q
0.5
0.0
0.0 0.2 (M 0.6
Northern plains killfish (Fundulus
Study
Birkner 1978
Birkner 1978
Birkner 1978
Birkner 1978
Birkner 1978
Site
PU
PU
TT
TT
TT
TT
TT
TT
o
e
^*^
jf^
0.8 1.0 1.2
kansae)
Site
3
11
23
27
30
CP
invert ^fish
0.61
0.61
1.07
1.07
1.07
1.07
1.07
1.07
Median ratio:
R2.
F:
df:
P:
r r
^ invert ^
3.10
5.65
15.50
34.60
45.05
0.93
1.40
2.63
2.88
2.68
1.91
1.90
1.80
2.04
0.61
9.24
6
0.015
fish
7.70
5.00
23.10
31.90
57.40
Ratio
1.52
2.30
2.47
2.70
2.51
1.79
1.78
1.69
Ratio
2.48
0.88
1.49
0.92
1.27
Do not distribute, quote or cite
B-112
Draft Document
-------
Northern plains
70 -
60 -
50 -
cflifc 40 "
JO
20 -
10 - Q,
1
0
killfish (Fundulus kansae)
Q
^
^^
^^
11111
10 20 30 40 50
C
^ in vert.
Median ratio:
R2:
F:
df:
P:
1.27
0.93
37.8
3
0.008
Rainbow trout (Oncorhynchus mykiss)
Study
Butler etal. 1993
Butler etal. 1993
Butler etal. 1993
Butler etal. 1995
Butler etal. 1995
Butler etal. 1995
Butler etal. 1995
Butler etal. 1994
Butler etal. 1997
Butler etal. 1997
Butler etal. 1997
Butler etal. 1997
Butler etal. 1997
Butler etal. 1997
Butler etal. 1997
Butler etal. 1997
Butler etal. 1997
Butler etal. 1997
Butler etal. 1997
Butler etal. 1997
Butler etal. 1997
Butler etal. 1994
Butler etal. 1994
Butler etal. 1993
Butler etal. 1991
Casey 2005
Site
LP2
LP2
LP3
MP
MP
MP
MP
NFK3
MN2
MN2
CHK
CHK
CHK
CHK
CHK
MN3
MN3
MN3
MN1
MN1
MN1
NFK2
NFK2
F2
4
Deerlick Cr.
C C
^^ invert ^fish
1.00
1.00
1.12
1.60
1.60
1.60
1.60
2.00
2.20
2.20
2.40
2.40
2.40
2.40
2.40
2.70
2.70
2.70
2.90
2.90
2.90
3.10
3.10
3.90
3.90
4.45
Ratio
1.27
1.40
1.90
2.10
2.30
2.50
3.06
4.70
2.10
2.80
2.20
2.29
2.50
2.80
2.90
2.60
3.69
4.90
2.50
2.60
3.20
3.60
35.68
7.60
3.50
2.09
1.27
1.40
1.70
1.31
1.44
1.56
1.91
2.35
0.95
1.27
0.92
0.96
1.04
1.17
1.21
0.96
1.37
1.81
0.86
0.90
1.10
1.16
11.51
1.95
0.90
0.47
Do not distribute, quote or cite
B-113
Draft Document
-------
Rainbow trout (Oncorhynchus mykiss)
Casey 2005
Butler etal. 1993
Butler etal. 1997
Low and Mullins 1990
Casey 2005
Casey 2005
Butler etal. 1994
Butler etal. 1994
40 -i
0
30 -
Cflsh 20 -
10 - „
J^
ft I* W
0
Red shiner (Cyprinella
Study
Butler etal. 1995
Butler etal. 1995
Butler etal. 1995
Butler etal. 1995
Butler etal. 1995
Butler etal. 1997
Butler etal. 1994
Butler etal. 1994
Butler etal. 1994
Lemly 1985
Mueller etal. 1991
Butler etal. 1991
Lemly 1985
May et al. 2008
Butler etal. 1994
May et al. 2008
Mueller etal. 1991
Deerlick Cr.
F2
WBR
5
Luscar Cr.
Luscar Cr.
HCC1
GUN2
0
8^-—***^***^^'^
__— — -*********^
0
1 1 1
10 20 30
*"- invert.
lutrensis)
Site
ME4
YJ2
SJ1
ME3
ME3
MN4
AD
LW
LSW1
Badin Lake
A3
3
High Rock Lake
SSW
IW
SSAU
A2
4.45
4.80
5.05
5.60
9.95
9.95
21.00
28.00
Median ratio:
R2:
F:
df:
P:
c c
^^ invert ^fish
1.55
1.65
2.50
2.55
2.55
2.65
2.70
3.00
3.90
5.18
6.00
6.20
6.75
7.60
8.35
8.35
8.50
3.34
7.60
5.10
2.60
11.16
13.71
26.76
5.40
1.19
0.16
6.22
32
0.005
Ratio
5.10
4.50
3.50
4.20
4.60
4.20
7.30
19.00
14.00
3.56
8.10
7.70
3.70
10.00
83.00
11.20
7.90
0.75
1.58
1.01
0.46
1.12
1.38
1.27
0.19
3.29
2.73
1.40
1.65
1.80
1.58
2.70
6.33
3.59
0.69
1.35
1.24
0.55
1.32
9.94
1.34
0.93
Do not distribute, quote or cite
B-114
Draft Document
-------
Red shiner (Cyprinella lutrensis)
Study
Butler etal. 1997
May et al. 2008
May et al. 2008
May et al. 2008
May et al. 2008
May et al. 2008
May et al. 2008
May et al. 2008
Lemly 1985
Site
MN5
NSK
sso
NSCU
NSCL
SSAL
KR
NSP
Belews Lake
Cr<
invert *^fish
8.60
8.81
10.00
10.50
10.70
11.50
17.20
24.00
45.53
Ratio
4.40
5.81
7.16
7.24
7.36
9.00
7.03
8.62
30.92
0.51
0.66
0.72
0.69
0.69
0.78
0.41
0.36
0.68
-fish
100 -|
80 -
60 -
40 -
20 -
0
o o
Median ratio: 1.28
R2:
F:
df:
P:
0.06
1.59
24
0.224
0 10 20 30
r*
'-"invert.
40
50
Not used because P > 0.05
Redside shiner (Richardsonius balteatus)
Study
Site
cfl;
ish
Ratio
Hamilton and Buhl 2004
Hamilton and Buhl 2004
Hamilton and Buhl 2005
ACM
LBR
BGS
6.70
7.70
10.80
6.00
2.70
13.20
0.90
0.35
1.22
Do not distribute, quote or cite
B-115
Draft Document
-------
Redside shiner (Richardsonius balteatus)
14 -1
12 -
10 -
CfiSh g . Q
4 - /
2 .
A
0 5
*- in vert
o
/
/
/
/
0
10
Median ratio:
R2:
F:
df:
P
15 Not used because P >
0.90
0.73
2.68
1
0.397
0.05
Roundtail chub (Gila robusta)
Study
Butler etal. 1994
Butler etal. 1994
Butler etal. 1994
Butler etal. 1994
Butler etal. 1994
Butler etal. 1994
Butler etal. 1994
Butler etal. 1994
Butler etal. 1994
Butler etal. 1994
Butler etal. 1995
Butler etal. 1997
Butler etal. 1994
Butler etal. 1994
Butler etal. 1994
Butler etal. 1994
Butler etal. 1994
Butler etal. 1991
Butler etal. 1994
Butler etal. 1994
Butler etal. 1993
Butler etal. 1994
Butler etal. 1994
Butler etal. 1997
Butler etal. 1994
Butler etal. 1994
Site
COL1
COL1
COL1
COL1
COL1
COL1
COL1
COL1
COL1
RB3
MP
MUD
AD
LW
NFK2
PSW1
LSW1
10
TGC
BSW1
F2
CRC
IW
NW2
RBI
GUN2
P P
^ invert ^fish
1.50
1.50
1.50
1.50
1.50
1.50
1.50
1.50
1.50
1.60
1.60
2.30
2.70
3.00
3.10
3.70
3.90
4.80
4.90
5.00
7.50
7.50
8.35
11.40
21.00
28.00
Ratio
2.20
2.50
2.70
3.30
3.70
4.10
5.10
5.30
26.00
5.40
4.20
4.60
7.10
4.50
6.10
7.70
5.80
1.90
10.00
8.10
7.30
19.00
8.50
6.90
5.90
6.80
1.47
1.67
1.80
2.20
2.47
2.73
3.40
3.53
17.33
3.38
2.63
2.00
2.63
1.50
1.97
2.08
1.49
0.40
2.04
1.62
0.97
2.53
1.02
0.61
0.28
0.24
Do not distribute, quote or cite
B-116
Draft Document
-------
Roundtail chub (Gila robustd)
30 -
25 -
20 -
Cfish 15
10 -
5 -
0 -
o
o
08 rP
ggU u O
B^ O
1 '
0 10 20
c
^ in vert.
Median ratio: 1.98
J
"Nfnt u«pr| hprai
1
30
R2: 0.01
F: 0.18
df: 24
P: 0.834
jse P > 0.05
Sand shiner (Notropis stramineus)
Study
GEI2013
GEI2013
GEI2013
GEI2013
GEI2013
GEI2013
GEI2013
GEI2013
GEI2013
GEI2013
GEI2013
GEI2013
GEI2013
GEI2013
GEI2013
GEI2013
GEI2013
GEI2013
GEI2013
GEI2013
GEI2013
GEI2013
GEI2013
GEI2013
Site
SW1
SW1
SW1
SW1
SW1
SW1
SW1
SW1
SW1
SW2-1
SW2-1
SW2-1
SW2-1
SW2-1
SWB
SWB
SWB
SWB
SW1
SW1
SW1
SW1
SW1
SW1
invert
6.54
6.54
6.54
6.54
6.54
6.54
6.54
6.54
6.54
6.60
6.60
6.60
6.60
6.60
7.06
7.06
7.06
7.06
7.82
7.82
7.82
7.82
7.82
7.82
Cflsh Ratio
8.43
9.02
9.66
11.21
11.85
11.94
13.50
14.05
14.14
18.70
19.33
19.77
20.39
23.70
8.27
9.01
9.81
10.22
11.33
12.05
12.22
12.55
12.65
12.68
1.29
1.38
1.48
1.71
1.81
1.83
2.06
2.15
2.16
2.84
2.93
3.00
3.09
3.59
1.17
1.28
1.39
1.45
1.45
1.54
1.56
1.60
1.62
1.62
Do not distribute, quote or cite
B-117
Draft Document
-------
Sand shiner (Notropis stramineus)
GEI2013
GEI2013
GEI2013
GEI2013
GEI2013
GEI2013
GEI2013
GEI2013
GEI2013
25 -\
20 -
15 -
cflsh io _
5 -
A
0 2
SW1
SW1
SW1
SW1
SW2-1
SW2-1
SW2-1
SW2-1
SW2-1
o
1
JU"
1^1
1 1 1
468
*"-- invert.
7.82
7.82
7.82
7.82
9.14
9.14
9.14
9.14
9.14
§
3
Median ratio:
R2:
F:
df:
P:
10
14.13
14.43
15.87
16.63
17.84
18.21
18.98
20.12
20.73
1.83
0.12
4.12
31
0.026
1.81
1.85
2.03
2.13
1.95
1.99
2.08
2.20
2.27
Sculpin (Cottoidea)
Study
Mason et al. 2000
Mason et al. 2000
Mason et al. 2000
Formation 2012
Formation 2012
Formation 2012
Formation 2012
Formation 2012
Formation 2012
Formation 2012
Formation 2012
Formation 2012
Formation 2012
Formation 2012
Formation 2012
Formation 2012
Site
BK
BK
BK
SFTC-1
SFTC-1
SFTC-1
CC-75
CC-75
CC-350
CC-350
SFTC-1
CC-75
CC-75
CC-75
CC-350
CC-150
c c
^invert ^fish
1.43
1.43
1.43
1.63
2.42
2.49
3.11
3.11
3.16
3.16
3.21
3.97
4.16
4.16
4.20
4.46
Ratio
1.16
2.35
2.64
9.31
5.68
5.87
5.03
5.58
6.47
7.12
3.75
3.77
7.08
7.19
5.28
5.04
0.81
1.64
1.84
5.71
2.35
2.36
1.62
1.79
2.05
2.26
1.17
0.95
1.70
1.73
1.26
1.13
Do not distribute, quote or cite
B-118
Draft Document
-------
Sculpin (Cottoidea)
Formation 2012
Formation 2012
Formation 2012
Formation 2012
Formation 2012
Formation 2012
Formation 2012
Formation 2012
Formation 2012
Formation 2012
Formation 2012
Formation 2012
Formation 2012
Formation 2012
Formation 2012
Formation 2012
Formation 2012
Formation 2012
Formation 2012
Formation 2012
Formation 2012
Formation 2012
Formation 2012
Formation 2012
Formation 2012
Formation 2012
Formation 2012
Formation 2012
Formation 2012
Formation 2012
Formation 2012
Formation 2012
Formation 2012
Formation 2012
Formation 2012
Formation 2012
Formation 2012
CC-150
CC-150
CC-3A
CC-3A
CC-3A
CC-150
DC-600
DC-600
DC-600
DC-600
DC-600
LSV-4
LSV-4
HS-3
HS-3
CC-350
CC-350
CC-1A
CC-1A
CC-1A
CC-1A
HS-3
CC-1A
CC-150
CC-3A
HS
HS
HS
LSV-2C
LSV-2C
HS-3
LSV-2C
HS-3
LSV-2C
HS
HS
LSV-2C
4.46
4.70
5.45
5.45
5.48
7.03
7.83
7.83
8.53
8.53
8.65
9.54
9.54
11.40
11.40
11.45
11.45
12.24
12.24
12.24
12.57
13.41
13.55
14.32
14.50
15.70
15.70
18.70
22.62
22.62
24.70
26.31
26.55
26.95
27.80
27.80
30.00
6.01
5.14
11.65
14.45
11.47
10.73
7.96
8.62
7.87
8.50
7.63
18.28
20.01
18.57
21.85
9.53
10.03
8.34
9.94
17.47
7.78
26.63
12.63
7.35
20.20
23.23
23.25
10.95
11.38
17.47
23.93
18.85
23.68
20.32
35.93
41.30
25.95
1.35
1.09
2.14
2.65
2.09
1.53
1.02
1.10
0.92
1.00
0.88
1.92
2.10
1.63
1.92
0.83
0.88
0.68
0.81
1.43
0.62
1.99
0.93
0.51
1.39
1.48
1.48
0.59
0.50
0.77
0.97
0.72
0.89
0.75
1.29
1.49
0.87
Do not distribute, quote or cite
B-119
Draft Document
-------
Sculpin (Cottoidea)
50
40
Q A
30
ftsh 20
10
0
• 0 LaJ^
@
0
Shorthead redhorse
Study
Roddy etal. 1991
Roddy etal. 1991
Roddy etal. 1991
Roddy etal. 1991
Roddy etal. 1991
Roddy etal. 1991
Roddy etal. 1991
Roddy etal. 1991
Roddy etal. 1991
Roddy etal. 1991
Roddy etal. 1991
4 -
3 -
Cflsll 2 -
1 -
n -
O
o
o ^»<-^6
Q O ^^>^U
(fiiP ° °
o
1 I 1 1
10 20 30 40
*- invert.
(Moxostoma macrolepidotum)
Site
18
18
18
18
18
18
18
18
18
18
18
1
Median ratio: 1.29
R2: 0.63
F: 87.0
df: 51
P: <0.001
Cjnvert Cflsh Ratio
3.10 2.80
3.10 2.90
3.10 2.90
3.10 3.10
3.10 3.30
3.10 3.40
3.10 3.50
3.10 3.60
3.10 3.70
3.10 3.80
3.10 3.80
Median ratio: 1.10
R2: 0.00
F: 0.00
df: 9
P: 1.0
0.90
0.94
0.94
1.00
1.06
1.10
1.13
1.16
1.19
1.23
1.23
Not used because P > 0.05
-'invert.
Do not distribute, quote or cite
B-120
Draft Document
-------
Smallmouth bass (Micropterm dolomieu)
Study
Butler etal. 1995
Butler etal. 1995
Butler etal. 1995
Butler etal. 1995
Butler etal. 1995
Butler etal. 1995
Butler etal. 1995
Butler etal. 1997
Mueller etal. 1991
Mueller etal. 1991
14.0
12.0
10,0
8.0
CF""' 6.0
•1.0
2.0 f=»^*~"~"
0.0
0.0 2.0
Site
MP
SU
SU
su
su
su
su
MNP3
Rl
Rl
o
^ ---~~'
^— ~~ — " o
--—""^ 0
4.0 6.0 8.0 10.0
CP
invert ^fish
1.60
1.85
1.85
1.85
1.85
1.85
1.85
6.15
8.70
8.70
Median ratio:
R2:
F:
df:
P:
Not used because P >
Ratio
.90
.50
.50
.48
.55
.72
.84
12.00
2.90
4.10
0.82
0.26
2.84
8
0.117
0.05
1.19
0.81
0.81
0.80
0.84
0.93
1.00
1.95
0.33
0.47
Speckled dace (Rhinichthys osculus)
Study
Hamilton and Buhl 2004
Butler etal. 1995
Butler etal. 1995
Butler etal. 1995
Butler etal. 1995
Butler etal. 1995
Butler etal. 1995
Butler etal. 1995
Butler etal. 1995
Butler etal. 1995
Butler etal. 1993
Butler etal. 1993
Butler etal. 1993
Butler etal. 1993
Site
use
AK
AK
AK
HD1
HD1
HD1
DD
DD
DD
LP3
Dl
Dl
Dl
^invert ^fish
0.50
0.78
0.78
0.78
0.83
0.83
0.83
0.86
0.86
0.86
1.12
1.20
1.20
1.20
Ratio
6.90
3.10
4.00
4.30
2.80
3.20
5.30
4.40
5.60
6.00
6.00
3.40
3.50
3.70
13.80
4.00
5.16
5.55
3.39
3.88
6.42
5.12
6.51
6.98
5.38
2.83
2.92
3.08
Do not distribute, quote or cite
B-121
Draft Document
-------
Speckled dace (Rhinichthys osculus)
Study
Butler
Butler
Butler
Butler
Butler
Butler
Butler
Butler
Butler
Butler
Butler
Butler
Butler
Butler
Butler
Butler
Butler
Butler
Butler
Butler
Butler
Butler
Butler
Butler
Butler
Butler
Butler
Butler
Butler
Butler
Butler
Butler
Butler
Butler
Butler
Butler
Butler
Butler
Butler
Butler
Butler
et
et
et
et
et
et
et
et
et
et
et
et
et
et
et
et
et
et
et
et
et
et
et
et
et
et
et
et
et
et
et
et
et
et
et
et
et
et
et
et
et
al.
al.
al.
al.
al.
al.
al.
al.
al.
al.
al.
al.
al.
al.
al.
al.
al.
al.
al.
al.
al.
al.
al.
al.
al.
al.
al.
al.
al.
al.
al.
al.
al.
al.
al.
al.
al.
al.
al.
al.
al.
1993
1993
1995
1993
1993
1993
1993
1994
1994
1994
1994
1994
1994
1994
1994
1994
1994
1995
1995
1995
1997
1993
1994
1993
1993
1993
1997
1997
1993
1997
1997
1997
1997
1993
1993
1993
1993
1993
1993
1995
1995
Site
Bl
Bl
ME2
B2
D2
D2
D2
COL1
COL1
COL1
COL1
COL1
COL1
COL1
COL1
COL1
RB3
YJ2
YJ2
YJ2
MNQ
PI
NFK3
SB1
SB1
SB1
MN2
MN2
ST1
MUD
MUD
CHK
CHK
Ul
Ul
Ul
Ul
Ul
Ul
SJ1
SJ1
invert
1
1
1
1
1
1
1
1
1
1
1
1
1
1
1
1
1
1
1
1
1
1
2
2
2
2
2
2
2
2
2
2
2
2
2
2
2
2
2
2
2
Cfish
.25
.25
.25
.35
.45
.45
.45
.50
.50
.50
.50
.50
.50
.50
.50
.50
.60
.65
.65
.65
.80
.95
.00
.15
.15
.15
.20
.20
.25
.30
.30
.40
.40
.45
.45
.45
.45
.45
.45
.50
.50
4
4
6
5
4
6
6
2
5
7
7
8
8
9
9
11
93
6
6
7
5
5
7
7
9
10
2
3
6
6
7
3
5
3
6
7
9
9
9
2
4
Ratio
.40
.40
.10
.80
.90
.50
.80
.30
.00
.30
.40
.40
.60
.30
.60
.00
.00
.30
.50
.10
.90
.50
.10
.80
.50
.00
.70
.60
.80
.10
.20
.80
.20
.60
.90
.30
.20
.40
.80
.90
.30
3
3
4
4
3
4
4
1
3
4
4
5
5
6
6
7
58
3
3
4
3
2
3
3
4
4
1
1
3
2
3
1
2
1
2
2
3
3
4
1
1
.52
.52
.88
.30
.38
.48
.69
.53
.33
.87
.93
.60
.73
.20
.40
.33
.13
.82
.94
.30
.28
.82
.55
.63
.42
.65
.23
.64
.02
.65
.13
.58
.17
.47
.82
.98
.76
.84
.00
.16
.72
Do not distribute, quote or cite
B-122
Draft Document
-------
Speckled dace (Rhinichthys osculus)
Study
Butler
Butler
Butler
Butler
Butler
Butler
Butler
Butler
Butler
Butler
Butler
Butler
Butler
Butler
Butler
Butler
Butler
Butler
Butler
Butler
Butler
Butler
Butler
Butler
Butler
Butler
Butler
Butler
Butler
Butler
Butler
Butler
Butler
Butler
Butler
Butler
Butler
Butler
Butler
Butler
Butler
et
et
et
et
et
et
et
et
et
et
et
et
et
et
et
et
et
et
et
et
et
et
et
et
et
et
et
et
et
et
et
et
et
et
et
et
et
et
et
et
et
al.
al.
al.
al.
al.
al.
al.
al.
al.
al.
al.
al.
al.
al.
al.
al.
al.
al.
al.
al.
al.
al.
al.
al.
al.
al.
al.
al.
al.
al.
al.
al.
al.
al.
al.
al.
al.
al.
al.
al.
al.
1995
1995
1995
1995
1997
1995
1997
1997
1993
1997
1993
1993
1993
1993
1993
1994
1994
1994
1994
1994
1994
1994
1994
1994
1994
1994
1994
1993
1993
1995
1993
1993
1994
1994
1993
1993
1993
1993
1994
1993
1991
Site
SJ1
ME3
ME3
ME3
MN4
MN1
MN3
MN3
SP2
MN1
SP1
SP1
SP1
WSB2
WSB2
LW
NFK2
NFK2
NFK2
NFK2
NFK2
NFK2
NFK2
NFK2
NFK2
NFK2
NFK2
LP4
ST2
ME1
WSB2
SB2
CF1
PSW1
SB2
SB2
R2
F2
LSW1
Rl
9
invert
2
2
2
2
2
2
2
2
2
2
2
2
2
3
3
3
3
3
3
3
3
3
3
3
3
3
3
3
3
3
3
3
3
3
3
3
3
3
3
4
4
Cfish
.50
.55
.55
.55
.65
.70
.70
.70
.75
.90
.95
.95
.95
.00
.00
.00
.10
.10
.10
.10
.10
.10
.10
.10
.10
.10
.10
.20
.35
.40
.60
.60
.60
.70
.75
.75
.90
.90
.90
.00
.10
5
2
5
7
7
5
4
6
12
3
7
7
8
6
7
62
4
5
5
6
6
6
6
6
6
7
8
8
9
6
11
12
6
13
7
10
6
8
83
8
5
Ratio
.10
.80
.50
.00
.90
.50
.30
.00
.00
.70
.00
.30
.90
.20
.60
.00
.80
.40
.70
.10
.20
.30
.40
.70
.90
.40
.70
.70
.30
.40
.70
.10
.10
.00
.80
.80
.00
.90
.00
.50
.70
2
1
2
2
2
2
1
2
4
1
2
2
3
2
2
20
1
1
1
1
2
2
2
2
2
2
2
2
2
1
3
3
1
3
2
2
1
2
21
2
1
.04
.10
.16
.75
.98
.04
.59
.22
.36
.28
.37
.47
.02
.07
.53
.67
.55
.74
.84
.97
.00
.03
.06
.16
.23
.39
.81
.72
.78
.88
.25
.36
.69
.51
.08
.88
.54
.28
.28
.13
.39
Do not distribute, quote or cite
B-123
Draft Document
-------
Speckled dace (Rhinichthys
Study
Butler etal. 1993
Butler etal. 1993
Butler etal. 1993
Butler etal. 1993
Butler etal. 1993
Butler etal. 1991
Butler etal. 1994
Butler etal. 1994
Butler etal. 1994
Butler etal. 1997
Butler etal. 1997
Butler etal. 1995
Hamilton and Buhl 2005
Butler etal. 1991
Hamilton and Buhl 2004
Butler etal. 1994
Hamilton and Buhl 2004
Butler etal. 1994
Butler etal. 1997
Hamilton and Buhl 2005
Butler etal. 1997
Hamilton and Buhl 2004
Butler etal. 1994
Butler etal. 1994
100 -|
80 -
60 -
Cflsh
20 -
n -
O
0
osculus)
Site
ST2
ST2
R2
ST2
WSB2
10
SMF
TGC
BSW1
WBR
WBR
NW
LiB
3
ACM
CRC
LBR
IW
MN5
SLC
NW2
DVC
LZA1
GUN2
invert
4
4
4
4
4
4
4
4
5
5
5
5
5
6
6
7
7
8
8
9
11
12
19
28
Cfish
10
10
30
50
75
80
80
90
00
05
05
10
40
20
70
50
70
35
60
70
40
80
00
00
Median ratio:
0
a^^>r££Lf
m&Sfj^
0
»__ •
*o^ o
R2:
F:
df:
P:
Ratio
8.50
10.70
17.10
15.70
15.60
4.80
7.80
12.00
15.00
5.50
9.70
8.70
5.80
6.50
8.50
13.00
5.60
10.00
14.00
15.20
11.00
7.50
28.00
8.90
2.79
0.01
1.71
118
0.185
2.07
2.61
3.98
3.49
3.28
1.00
1.63
2.45
3.00
1.09
1.92
1.71
1.07
1.05
1.27
1.73
0.73
1.20
1.63
1.57
0.96
0.59
1.47
0.32
10
r
^-invert.
20
30
Not used because P > 0.05
Do not distribute, quote or cite
B-124
Draft Document
-------
Sucker (Catostomidae)
Study
Butler etal. 1995
Butler etal. 1995
Butler etal. 1993
Rinella and Schuler 1992
Butler etal. 1993
Butler etal. 1995
Butler etal. 1993
Butler etal. 1993
Butler etal. 1993
Butler etal. 1991
Butler etal. 1994
Butler etal. 1993
Butler etal. 1993
Butler etal. 1993
Butler etal. 1993
Butler etal. 1993
Butler etal. 1993
40 -j
0
30 -
C">n -
fish "-u
10 -
_«-*— -o ^
0 2
*"-• invert.
Site
HD2
HD2
Dl
Malheur Lake
B2
YJ2
PI
Ul
Ul
12
NFK2
SB2
R2
R2
ST2
WSB2
F2
0°^^— — —
5 — " — '
O
468
Cp
invert ^fish
0.98
0.98
1.20
1.20
1.35
1.65
1.95
2.45
2.45
2.80
3.10
3.60
3.90
4.30
4.50
4.75
7.50
Median ratio:
R2:
F:
df:
P:
Not used because P >
Ratio
0.68
0.76
2.30
1.60
1.80
2.20
1.50
2.30
3.60
2.10
35.00
5.10
5.00
2.20
10.00
11.80
4.20
1.33
0.07
1.10
15
0.360
0.05
0.69
0.78
1.92
1.33
1.33
1.33
0.77
0.94
1.47
0.75
11.29
1.42
1.28
0.51
2.22
2.48
0.56
Sunfish (Centrarchidae)
Study
Welsh and Maughan 1994
Welsh and Maughan 1994
Welsh and Maughan 1994
Welsh and Maughan 1994
Welsh and Maughan 1994
Welsh and Maughan 1994
Welsh and Maughan 1994
Site
outfall drain
Pretty Water
Hart Mine Marsh
outfall drain
outfall drain
Pretty Water
Old Channel
c c
^^ invert ^fish
1.15
1.16
1.20
1.30
1.30
1.50
1.50
Ratio
2.30
1.80
2.40
2.10
2.80
1.60
2.00
2.00
1.56
2.00
1.62
2.15
1.07
1.33
Do not distribute, quote or cite
B-125
Draft Document
-------
Sunfish (Centrarchidae)
Welsh and M
Welsh and M
Welsh and M
Welsh and M
Welsh and M
GEI2013
12
10
8
Cflsh 6
4
2
0
aughan 1994 Pretty Water 1.50 2.30 1.53
aughan 1994 CibolaLake 1.85 5.90 3.19
aughan 1994 CibolaLake 1.90 5.30 2.79
aughan 1994 CibolaLake 1.90 7.60 4.00
aughan 1994 Oxbow Lake 3.60 11.00 3.06
SW2-1 9.14 8.10 0.89
o
^^^ _. Median ratio: 2.00
0 ^^^^ °
%^^^^ R2: 0.38
^^ F: 6.66
G& df: 11
P- nnp
0 2 4 6 8 10
invert.
Tui chub (Gila bicolor)
Study Site Cinvert Cflsh Ratio
Sorenson & 5
Sorenson & J
Rinella and S
4 -
3 -
3 -
2 -
1 -
1 -
>chwarzbach 1991 5 0.49 1.20 2.45
Schwarzbach 1991 4 0.76 1.00 1.32
chuler!992 Harney Lake 2.05 3.10 1.51
^^ Median ratio: 1.51
.s^ R2: 0.94
0.05
^ ID vert.
Do not distribute, quote or cite
B-126
Draft Document
-------
Walleye (Sander vitreus)
Study
Site
Butler etal. 1995 PU
Butler etal. 1995 PU
Butler etal. 1995 PU
Butler etal. 1995 PU
Butler etal. 1995 PU
Butler etal. 1995 TT
Butler etal. 1995 TT
Butler etal. 1995 TT
Butler etal. 1995 TT
Butler etal. 1995 TT
Butler etal. 1995 TT
Peterson et al
Peterson et al
Peterson et al
Peterson et al
Mueller et al.
10 -1
R -
O
6 -
Cflsh
7 -
n -
1991 7
1991 7
1991 7
1991 7
1991 Rl
0
0
^ -~
JL-~""""~ —
a—-—" — """""""
|i^ °
e
invert
0
0
0
0
0
1
1
1
1
1
1
3
3
3
3
8
Cfish
61
61
61
61
61
07
07
07
07
07
07
83
83
83
83
70
Median ratio:
0 2 4 6 8 10
c^
'-"invert.
R2:
F:
df:
P:
Ratio
0.89
.00
.27
.66
.72
.60
.86
2.00
2.55
2.68
2.68
4.27
4.79
6.76
8.35
2.40
1.82
0.24
4.46
14
0.032
1
1
2
2
2
1
1
1
2
2
2
1
1
1
2
0
46
64
09
72
82
50
75
88
39
51
51
11
25
77
18
28
Western mosquitofish (Gambusia affinis)
Study
Site
Saiki etal. 1993 ET6
Saiki etal. 1993 ET6
Saiki etal. 1993 ET7
Saiki etal. 1993 ET7
Saiki etal. 1993 SJR1
Saiki etal. 1993 SJR1
Saiki and Lowe 1987 Volta Pond 26
^invert
0
0
0
0
0
0
1
Cfish
85
85
86
86
95
95
42
Ratio
1.00
1.30
0.90
1.00
0.95
1.30
1.24
1
1
1
1
1
1
0
18
54
05
16
01
38
87
Do not distribute, quote or cite
B-127
Draft Document
-------
Western mosquitofish
Saiki and Lowe 1987
Saikietal. 1993
Saikietal. 1993
Saiki and Lowe 1987
Saiki and Lowe 1987
GEI2013
GEI2013
GEI2013
GEI2013
GEI2013
Saikietal. 1993
Saikietal. 1993
GEI2013
Saikietal. 1993
Saikietal. 1993
Saikietal. 1993
Saikietal. 1993
Saiki and Lowe 1987
Saiki and Lowe 1987
Saiki and Lowe 1987
Saiki and Lowe 1987
Saiki and Lowe 1987
Saiki and Lowe 1987
Saiki and Lowe 1987
Saiki and Lowe 1987
350 -i
300 -
250 -
200 -
fish 15Q .
100 -
50 \^<>^*'
^ J&"*^
(Gambusia a/finis)
Volta Pond 26
SJR3
SJR3
Volta Wasteway
Volta Wasteway
SWA1
SWA1
SWA1
SWA1
SWA1
SJR2
SJR2
SWA1
GT4
GT4
GT5
GT5
Kesterson Pond 1 1
Kesterson Pond 1 1
Kesterson Pond 8
Kesterson Pond 8
Kesterson Pond 2
Kesterson Pond 2
San Luis Drain
San Luis Drain
o
o ^-"•"o
^^
0 ^***^ °
J^
i i i i
1.42
1.50
1.50
2.23
2.23
2.81
2.81
2.81
2.81
2.81
3.30
3.30
3.64
4.05
4.05
4.90
4.90
60.65
60.65
102.50
102.50
177.00
177.00
190.00
190.00
Median ratio:
R2:
F:
df:
P:
1.28
1.70
2.00
1.35
1.36
3.01
3.49
3.66
3.89
4.27
2.20
4.50
2.91
4.50
4.90
11.00
16.00
104.00
130.00
164.00
223.00
224.00
247.00
149.00
332.00
1.25
0.90
256.4
30
<0.001
0.90
1.13
1.33
0.61
0.61
1.07
1.24
1.30
1.39
1.52
0.67
1.36
0.80
1.11
1.21
2.24
3.27
1.71
2.14
1.60
2.18
1.27
1.40
0.78
1.75
0 50 100 150 200
invert.
Do not distribute, quote or cite
B-128
Draft Document
-------
Western cutthroat trout (Oncorhynchus clarkii lewisi)
Study Site
Minnow 2007 BA6
Minnow 2007 AL4
Minnow 2007 MI5
Minnow 2007 EL 12
Minnow 2007 EL 14
Minnow 2007 FO9
Minnow 2007 MI3
Minnow 2007 MI2
Minnow 2007 ELI
Minnow 2007 LI8
Minnow 2007 FO10
Minnow 2007 HA7
Minnow 2007 CL11
70 -1
60 - 0
50 " o ^
C 4° Jxx'X^
fish 30 . ^^^
20 - .^ °
10 - <^$
0 10 20 30
r*
'-"invert.
Cjnvert Cflsh Ratio
3.27 6.98 2.13
3.92 4.44 1.13
4.00 5.12 1.28
4.01 7.42 1.85
4.41 4.52 1.02
4.44 7.80 1.76
6.21 5.65 0.91
6.69 5.16 0.77
7.08 4.82 0.68
7.81 9.36 1.20
17.51 45.94 2.62
22.41 21.10 0.94
30.87 57.27 1.86
Median ratio: 1.20
R2: 0.81
F: 47.6
df: 11
P: <0.001
40
White sucker (Catostomus commersonii)
Study Site
Butler etal. 1993 LP3
Butler etal. 1993 Bl
Butler etal. 1993 D2
Butler etal. 1993 D2
Butler etal. 1993 PI
Butler etal. 1993 PI
Butler etal. 1995 MP
Butler etal. 1995 SU
Grasso etal. 1995 17
Grasso etal. 1995 17
Grasso etal. 1995 17
Cjnvert Cflsh Ratio
1.12 2.50 2.24
1.25 2.60 2.08
1.45 1.90 1.31
1.45 2.50 1.72
1.50 1.70 1.13
1.50 1.80 1.20
1.60 1.40 0.88
1.85 1.20 0.65
1.91 2.84 1.49
1.91 3.19 1.67
1.91 3.44 1.80
Do not distribute, quote or cite
B-129
Draft Document
-------
White sucker (Catostomus commersonii)
Study
Grassoetal. 1995
Grassoetal. 1995
Grassoetal. 1995
Butler etal. 1994
Butler etal. 1993
Lambing etal. 1994
Mueller et al. 1991
GEI2013
GEI2013
GEI2013
Mason et al. 2000
Mason et al. 2000
Mason et al. 2000
Butler etal. 1993
Butler etal. 1993
Butler etal. 1993
GEI2013
GEI2013
GEI2013
GEI2013
GEI2013
GEI2013
GEI2013
GEI2013
GEI2013
GEI2013
Butler etal. 1993
GEI2013
GEI2013
GEI2013
GEI2013
GEI2013
GEI2013
GEI2013
GEI2013
GEI2013
GEI2013
GEI2013
GEI2013
GEI2013
GEI2013
Site
17
17
17
NFK3
ST1
S33
Al
SWA1
SWA1
SWA1
HCRT
HCRT
HCRT
WSB2
SP2
LP4
SW4-
SW4-
SW4-
SW4-
SW4-
SW4-
SW4-
SW4-
SW4-
SW4-
ST2
LG1
LG1
LG1
LG1
LG1
LG1
LG1
LG1
LG1
LG1
LG1
LG1
LG1
LG1
Cr<
invert *^fish
1.91
1.91
1.91
2.00
2.25
2.40
2.70
2.81
2.81
2.81
2.81
2.81
2.81
3.00
3.15
3.20
3.33
3.33
3.33
3.33
3.33
3.33
3.33
3.33
3.33
3.33
3.35
3.37
3.37
3.37
3.37
3.37
3.39
3.56
3.56
3.56
3.56
3.56
3.56
3.56
3.56
Ratio
3.64
4.00
4.01
3.90
4.90
3.50
4.20
2.83
3.89
4.18
0.81
1.43
1.43
3.90
3.50
2.80
3.01
3.45
3.50
3.62
4.04
4.08
4.13
4.17
4.34
4.78
7.00
3.54
3.55
3.90
3.95
4.48
3.00
2.72
2.80
2.89
2.99
3.04
3.08
3.13
3.18
1.91
2.09
2.10
1.95
2.18
1.46
1.56
1.01
1.39
1.49
0.29
0.51
0.51
1.30
1.11
0.88
0.91
1.04
1.05
1.09
1.22
1.23
1.24
1.25
1.31
1.44
2.09
1.05
1.05
1.16
1.17
1.33
0.88
0.77
0.79
0.81
0.84
0.86
0.87
0.88
0.89
Do not distribute, quote or cite
B-130
Draft Document
-------
White sucker (Catostomus commersonii)
Study
GEI2013
GEI2013
Butler etal. 1993
Butler etal. 1993
GEI2013
GEI2013
GEI2013
GEI2013
GEI2013
GEI2013
Butler etal. 1993
Butler etal. 1993
Butler etal. 1993
GEI2013
GEI2013
GEI2013
GEI2013
GEI2013
Peterson et al. 1991
Peterson et al. 1991
Butler etal. 1993
Butler etal. 1991
GEI2013
GEI2013
Butler etal. 1993
Butler etal. 1993
GEI2013
GEI2013
GEI2013
GEI2013
GEI2013
GEI2013
GEI2013
GEI2013
GEI2013
GEI2013
GEI2013
GEI2013
GEI2013
GEI2013
GEI2013
Site
LG1
LG1
WSB2
WSB2
SWA1
SWA1
SWA1
SWA1
SWA1
SWA1
SB2
R2
SB2
CC1
CC1
CC1
CC1
CC1
7
7
R2
4
SW88
SW88
Rl
ST2
SW9
SW9
SW9
SW9
SW9
SW9
CC1
CC1
CC1
CC1
CC1
CC1
CC1
CC1
CC1
Cr<
invert *^fish
3.56
3.56
3.60
3.60
3.64
3.64
3.64
3.64
3.64
3.64
3.65
3.70
3.75
3.76
3.76
3.76
3.76
3.76
3.83
3.83
3.90
3.90
3.96
3.96
4.00
4.10
4.45
4.45
4.45
4.45
4.45
4.45
4.69
4.69
4.69
4.69
4.69
4.69
4.69
4.69
4.69
Ratio
3.25
3.27
4.30
6.30
2.83
3.39
3.47
3.55
3.63
3.75
4.30
4.20
4.80
5.99
6.56
7.21
7.42
7.62
3.30
4.64
5.40
5.30
4.63
4.75
9.50
8.30
4.07
4.18
4.19
4.20
4.40
5.18
4.51
4.57
4.94
5.02
5.81
6.01
6.43
7.25
8.00
0.91
0.92
1.19
1.75
0.78
0.93
0.95
0.98
1.00
1.03
1.18
1.14
1.28
1.59
1.74
1.92
1.97
2.03
0.86
1.21
1.38
1.36
1.17
1.20
2.38
2.02
0.91
0.94
0.94
0.94
0.99
1.16
0.96
0.98
1.05
1.07
1.24
1.28
1.37
1.55
1.71
Do not distribute, quote or cite
B-131
Draft Document
-------
White sucker (Catostomus commersonii)
Study
GEI2013
Butler etal. 1993
GEI2013
GEI2013
GEI2013
GEI2013
GEI2013
GEI2013
GEI2013
GEI2013
GEI2013
GEI2013
Butler etal. 1991
GEI2013
GEI2013
GEI2013
GEI2013
GEI2013
GEI2013
GEI2013
GEI2013
GEI2013
GEI2013
GEI2013
GEI2013
GEI2013
GEI2013
GEI2013
GEI2013
Butler etal. 1994
Mueller et al. 1991
GEI2013
GEI2013
GEI2013
GEI2013
GEI2013
Lambing etal. 1994
Lambing etal. 1994
Lambing etal. 1994
Butler etal. 1994
Butler etal. 1994
Site
CC1
F2
CC1
CC1
CC1
CC1
CC1
CC1
CC1
CC1
CC1
CC1
3
SWB
SWB
SWB
SWB
SWB
SWB
SWB
SWB
SWB
SWB
SWB
SWB
SWB
SWB
SWB
SWB
IW
Rl
SW2-1
SW2-1
SW2-1
SW2-1
SW2-1
S34
S34
S34
HCC1
GUN2
Cr<
invert *^fish
4.69
4.80
5.86
5.86
5.86
5.86
5.86
5.86
5.86
5.86
5.86
5.86
6.20
7.06
7.06
7.06
7.06
7.06
7.06
7.06
7.06
7.44
7.44
7.44
7.44
7.44
7.44
7.44
7.44
8.35
8.70
9.14
9.14
9.14
9.14
9.14
14.00
14.00
14.00
21.00
28.00
Ratio
8.52
5.20
5.00
5.37
5.59
5.71
5.90
6.61
6.79
6.82
7.29
7.48
1.80
7.18
7.36
7.98
8.03
9.65
12.76
12.85
13.16
8.21
8.77
8.85
9.87
10.97
13.59
15.75
16.40
9.70
3.40
16.54
18.14
18.54
19.16
21.29
25.30
28.00
29.00
3.00
20.00
1.82
1.08
0.85
0.92
0.95
0.98
1.01
1.13
1.16
1.16
1.25
1.28
0.29
1.02
1.04
1.13
1.14
1.37
1.81
1.82
1.86
1.10
1.18
1.19
1.33
1.48
1.83
2.12
2.21
1.16
0.39
1.81
1.99
2.03
2.10
2.33
1.81
2.00
2.07
0.14
0.71
Do not distribute, quote or cite
B-132
Draft Document
-------
White sucker (Catostomus commersonii)
Study
Butler etal. 1991
50 -I
40 -
30 -
Cflsh 20 -
|
10 - AJ
n *^^r®
0
Site
7
0
8 ^
\j jrf***^
i ^^^°
go ^^^--^
r^ o
0 0
10 20 30
r*
'-"invert.
Cp
invert ^fish
29.80
Median ratio:
R2:
F:
df:
P
40
Ratio
7.90
1.18
0.49
129.9
134
<0.001
0.27
Yellow perch (Percaflavescens)
Study
Butler etal. 1995
Butler etal. 1995
Butler etal. 1995
Butler etal. 1995
Butler etal. 1995
Butler etal. 1995
Belize et al. 2006
Belize et al. 2006
Belize et al. 2006
Belize et al. 2006
Peterson et al. 1991
Belize et al. 2006
Belize et al. 2006
Belize et al. 2006
Belize et al. 2006
GEI2013
GEI2013
GEI2013
GEI2013
GEI2013
Lambing etal. 1994
Site
PU
TT
TT
MP
MP
MP
Halfway
Geneva
Bethel
McFarlane
7
Long
Ramsey
Windy
Nelson
SW11
SW11
SW11
SW11
SW11
S34
c c
^^ invert ^fish
0.61
1.07
1.07
1.60
1.60
1.60
1.74
2.29
2.61
3.79
3.83
4.42
4.97
6.32
6.79
8.41
8.41
8.41
8.41
8.41
14.00
Ratio
1.10
1.60
1.70
2.00
2.20
2.70
2.72
3.30
3.09
5.40
7.33
6.28
7.64
6.06
10.68
4.54
5.49
5.50
5.58
5.68
67.00
1.80
1.50
1.60
1.25
1.38
1.69
1.56
1.44
1.19
1.42
1.91
1.42
1.54
0.96
1.57
0.54
0.65
0.65
0.66
0.68
4.79
Do not distribute, quote or cite
B-133
Draft Document
-------
Yellow perch (Perca flavescens)
Study
Site
Cinvert
Cfl
lsh
Ratio
80 -1
60 -
Cflsh 40 -
20 -
0 -
0
Median ratio: 1.42
^^~-~- R2: 0.46
^^—~-""~""^ F: 16.24
ile®"'"""c?3~Q df: 19
jT\t^CsfJ^r*^^if "
P: <0.001
0 5 10 15
1^
'-"invert.
Do not distribute, quote or cite
B-134
Draft Document
-------
Table B-7. Final vertebrate Trophic Transfer Factor (TTF) values, including estimated values using taxonomic classification.
Order Family Genus TTF TTF source data
Common name
Scientific name
alligator gar
black bullhead
black crappie
black redhorse
blacknose dace
blue catfish
bluegill
bluehead sucker
brassy minnow
brook stickleback
brook trout
brown bullhead
brown trout
bullhead
chain pickerel
channel catfish
common carp
common snook
crappie
creek chub
cutthroat trout
dolly varden
fathead minnow
flannelmouth sucker
flathead catfish
flathead chub
freshwater drum
gizzard shad
goldeye
green sunfish
iowa darter
kokanee salmon
largemouth bass
largescale sucker
longnose dace
longnose sucker
Do not distribute, quote
Atractosteus spatula
Ameiurus melas
Pomoxis nigromaculatus
Moxostoma duquesnei
Rhinichthys atratulus
Ictalurus furcatus
Lepomis macrochims
Catostomus discobolus
Hybognathus hankinsoni
Culaea inconstans
Salvelinus fontinalis
Ameiurus nebulosus
Salmo trutta
Esox niger
Ictalurus punctatus
Cyprinus carpio
Centropomus undecimalis
Pomoxis sp.
Semotilus atromaculatus
Oncorhynchus clarkii
Salvelinus malma
Pimephales promelas
Catostomus latipinnis
Pylodictis olivaris
Platygobio gracilis
Aplodinotus grunniens
Dorosoma cepedianum
Hiodon alosoides
Lepomis cyanellus
Etheostoma exile
Oncorhynchus nerka
Microptems salmoides
Catostomus macrocheilus
Rhinichthys cataractae
Catostomus catostomus
or cite
Lepistosteiformes
Siluriformes
Perciformes
Cypriniformes
Cypriniformes
Siluriformes
Perciformes
Cypriniformes
Cypriniformes
Gasterosteiformes
Salmo niformes
Siluriformes
Salmo niformes
Siluriformes
Esociformes
Siluriformes
Cypriniformes
Perciformes
Perciformes
Cypriniformes
Salmo niformes
Salmo niformes
Cypriniformes
Cypriniformes
Siluriformes
Cypriniformes
Perciformes
Clupeiformes
Hiodontiformes
Perciformes
Perciformes
Salmo niformes
Perciformes
Cypriniformes
Cypriniformes
Cypriniformes
Lepisosteidae
Ictaluridae
Centrarchidae
Catostomidae
Cyprinidae
Ictaluridae
Centrarchidae
Catostomidae
Cyprinidae
Gasterosteidae
Salmonidae
Ictaluridae
Salmonidae
Ictaluridae
Esocidae
Ictaluridae
Cyprinidae
Centropomidae
Centrarchidae
Cyprinidae
Salmonidae
Salmonidae
Cyprinidae
Catostomidae
Ictaluridae
Cyprinidae
Sciaenidae
Clupeidae
Hiodontidae
Centrarchidae
Percidae
Salmonidae
Centrarchidae
Catostomidae
Cyprinidae
Catostomidae
B-135
Atractosteus
Ameiurus
Pomoxis
Moxostoma
Rhinichthys
Ictalurus
Lepomis
Catostomus
Hybognathus
Culaea
Salvelinus
Ameiurus
Salmo
Esox
Ictalurus
Cyprinus
Centropomus
Pomoxis
Semotilus
Oncorhynchus
Salvelinus
Pimephales
Catostomus
Pylodictus
Platygobio
Aplodinotus
Dorosoma
Hiodon
Lepomis
Etheostoma
Oncorhynchus
Micropterus
Catostomus
Rhinichthys
Catostomus
1.27
0.91
2.67
1.05
1.46
0.73
1.48
1.04
1.46
1.69
0.88
0.91
1.44
0.82
2.04
0.73
1.34
1.48
2.67
1.12
1.07
0.88
1.57
1.06
0.82
1.46
1.48
1.27
1.27
1.27
1.62
1.19
1.27
1.05
1.46
0.90
All fish
Exact match
Exact match
Family Catostomidae
Family Cyprinidae
Genus Ictalurus
Exact match
Exact match
Family Cyprinidae
Exact match
Exact match
Genus Ameiurus
Exact match
Family Ictaluridae
Genus Esox
Exact match
Exact match
Order Perciformes
Genus Pomoxis
Exact match
Exact match
Genus Salvelinus
Exact match
Exact match
Family Ictaluridae
Family Cyprinidae
Order Perciformes
All fish
All fish
Exact match
Family Percidae
Genus Oncorhynchus
Exact match
Genus Catostomus
Family Cyprinidae
Exact match
Draft Document
-------
Common name
Scientific name
Order
Family
Genus
TTF
TTF source data
mixed
mosquitofish
mottled sculpin
mountain whitefish
northern pike
northern pikeminnow
northern plains killifish
northern redbelly dace
northern squawfish
quillback
rainbow trout
razorback sucker
red shiner
redbreast sunfish
redear sunfish
redside shiner
river carpsucker
river redhorse
rock bass
roundtail chub
sacramento perch
sacramento pikeminnow
sailfin molly
sand shiner
sauger
sculpin
shadow bass
shorthead redhorse
silver carp
smallmouth bass
smallmouth buffalo
speckled dace
spotted bass
spotted gar
stonecat
striped bass
striped mullet
sucker
Gambusia sp.
Cottus bairdi
Prosopium williamsoni
Esox lucius
Ptychocheilus oregonensis
Fundulus kansae
Chrosomus eos
Ptychocheilus oregonensis
Carpiodes cyprinus
Oncorhynchus mykiss
Xyrauchen texanus
Cyprinella lutrensis
Lepomis auritus
Lepomis microlophus
Richardsonius balteatus
Carpiodes carpio
Moxostoma carinatum
Ambloplites rupestris
Gila robusta
Archoplites interruptus
Ptychocheilus grandis
Poecilia latipinna
Notropis stramineus
Sander canadensis
Cottus sp.
Ambloplites ariommus
Moxostoma macrolepidotum
Hypophthalmichthys molitrix
Micropterus dolomieu
Ictiobus bubalus
Rhinichthys osculus
Micropterus punctulatus
Lepisosteus oculatus
Noturus flavus
Morone saxatilis
Mugil cephalus
Cyprinodontiformes
Scorpaeniformes
Salmoniformes
Esociformes
Cypriniformes
Cyprinodontiformes
Cypriniformes
Cypriniformes
Cypriniformes
Salmoniformes
Cypriniformes
Cypriniformes
Perciformes
Perciformes
Cypriniformes
Cypriniformes
Cypriniformes
Perciformes
Cypriniformes
Perciformes
Cypriniformes
Cyprinodontiformes
Cypriniformes
Perciformes
Scorpaeniformes
Perciformes
Cypriniformes
Cypriniformes
Perciformes
Cypriniformes
Cypriniformes
Perciformes
Lepistosteiformes
Siluriformes
Perciformes
Mugiliformes
Cypriniformes
Poeciliidae
Cottidae
Salmonidae
Esocidae
Cyprinidae
Fundulidae
Cyprinidae
Cyprinidae
Catostomidae
Salmonidae
Catostomidae
Cyprinidae
Centrarchidae
Centrarchidae
Cyprinidae
Catostomidae
Catostomidae
Centrarchidae
Cyprinidae
Centrarchidae
Cyprinidae
Poeciliidae
Cyprinidae
Percidae
Cottidae
Centrarchidae
Catostomidae
Cyprinidae
Centrarchidae
Catostomidae
Cyprinidae
Centrarchidae
Lepisosteidae
Ictaluridae
Moronidae
Mugilidae
Catostomidae
Gambusia
Cottus
Prosopium
Esox
Ptychocheilus
Fundulus
Chrosomus
Ptychocheilus
Carpiodes
Oncorhynchus
Xyrauchen
Cyprinella
Lepomis
Lepomis
Richardsonius
Carpiodes
Moxostoma
Ambloplites
Gila
Archoplites
Ptychocheilus
Poecilia
Notropis
Sander
Cottus
Ambloplites
Moxostoma
Hypophthalmichthys
Micropterus
Ictiobus
Rhinichthys
Micropterus
Lepisosteus
Noturus
Morone
Mugil
0.87 Exact match
0.86 Exact match
1.38 Exact match
1.38 Exact match
2.04 Exact match
1.46 Family Cyprinidae
1.27 Exact match
1.46 Family Cyprinidae
1.46 Family Cyprinidae
1.05 Family Catostomidae
1.19 Exact match
1.05 Family Catostomidae
1.46 Family Cyprinidae
1.37 Genus Lepomis
1.37 Genus Lepomis
1.46 Family Cyprinidae
1.05 Family Catostomidae
1.05 Family Catostomidae
1.48 Family Centrarchidae
1.46 Family Cyprinidae
1.48 Family Centrarchidae
1.46 Family Cyprinidae
1.06 Family Poeciliidae
1.83 Exact match
1.82 Genus Sander
1.29 Exact match
1.48 Family Centrarchidae
1.05 Family Catostomidae
1.46 Family Cyprinidae
1.27 Genus Micropterus
1.05 Family Catostomidae
1.46 Family Cyprinidae
1.27 Genus Micropterus
1.27 All fish
0.82 Family Ictaluridae
1.48 Order Perciformes
1.27 All fish
1.05 Family Catostomidae
Do not distribute, quote or cite
B-136
Draft Document
-------
Common name
Scientific name
Order
Family
Genus
TTF
TTF source data
sunfish species
tilapia
trout species
tui chub
utah sucker
walleye
western mosquitofish
westslope cutthroat trout
white bass
white crappie
white sturgeon
white sucker
wiper
yellow perch
Oncorhynchus sp.
Gila bicolor
Catostomus ardens
Sander vitreus
Gambusia affinis
Oncorhynchus clarkii lewisi
Morone chrysops
Pomoxis annularis
Acipenser transmontanus
Catostomus commersonii
Morone chrysops x Moron saxatilis
Percaflavescens
Perciformes
Perciformes
Salmoniformes
Cypriniformes
Cypriniformes
Perciformes
Cyprinodontiformes
Salmoniformes
Perciformes
Perciformes
Acipenseriformes
Cypriniformes
Perciformes
Perciformes
Centrarchidae 2.00 Exact match
Cichlidae 1.48 Order Perciformes
Salmonidae Oncorhynchus 1.19 Genus Oncorhynchus
Cyprinidae Gila 1.46 Family Cyprinidae
Catostomidae Catostomus 1.05 Genus Catostomus
Percidae Sander 1.82 Exact match
Poeciliidae Gambusia 1.25 Exact match
Salmonidae Oncorhynchus 1.20 Exact match
Moronidae Morone 1.48 Order Perciformes
Centrarchidae Pomoxis 2.67 Genus Pomoxis
Acipenseridae Acipenser 1.27 All fish
Catostomidae Catostomus 1.18 Exact match
Moronidae Morone 1.48 Order Perciformes
Percidae Perca 1.42 Exact match
Do not distribute, quote or cite
B-137
Draft Document
-------
FOOD WEB MODELS USED TO CALCULATE COMPOSITE TTFS TO TRANSLATE THE EGG-OVARY FCV TO WATER-COLUMN VALUES.
Table B-8. Food web
Referen Site Site
ce descript ID
ion
Default
Default
Default
Default
Default
Default
Default
models used to
Target Fish
fish TTF
species
commo
n name
black 0.
bullhea
d
black 2.
crappie
blackno 1 .
se dace
blue 0.
catfish
bluegill 1.
bluehea 1 .
d
sucker
brassy 1 .
minnow
calculate composite TTFs to translate the egg-ovary FCV to
Fish prey as described in NatureServe Fish prey spp 1° TL2 TTF
comment species used
91 Omnivorous bottom feeder; often eats
aquatic insects, crustaceans, molluscs,
occasionally fishes and carrion
67 Primarily a midwater feeder; zooplankton
and small Diptera larvae predominate in
the diet of individuals to 12 cm SL, while
fishes and aquatic insects predominate in
the diet of larger individuals
46 Eats immature aquatic insects,
amphipods, and various other aquatic
invertebrates; also eats algae and
diatoms, which may be of little
nutritional value (Smith 1979, Becker
1983).
73 Bottom feeder. Eats mostly crustaceans
and aquatic insects when young. Later,
fishes and large invertebrates become
most important (Moyle 1976). Also
scavenges.
48 Feeds opportunistically on aquatic insect
larvae, planktonic crustaceans, flying
insects, snails, and other small
invertebrates; small fishes, fish eggs,
crayfish, and algae sometimes are eaten.
Larvae and juveniles often eat
cladocerans and copepod nauplii. Adults
eats mainly aquatic insects, crayfishes,
and small fishes, or, in some bodies of
water, mostly zooplankton. Feeds at all
levels of water column.
04 Herbivore, Invertivore
46 Eats algae, phyto- and zooplankton,
benthic invertebrates, surface drift,
Median of all
insects
Median of all
insects
Median of all
insects
Median of all
insects
Median of all
insects
TL1
TL1
a water-column value at aquatic sites where sufficient data was available to calculate an enrichement factor (EF).
1°TL2 1° 1°TL2 2°TL2 2° 2° 2° TL2 3° TL2 3° 3° 3° TL2 4° TL2 4° 4° 4° TL2
spp TL2 proportio spp used TL2 TL2 proportio spp used TL2 TL2 proportio spp used TL2 TL2 proportio
abbrev TT n spp TT n spp TT n spp TT n
F abrev F abrev F abrev F
in 2.14 0.45 Median of crs 1.41 0.45 Median of bvs 4.29 0.10
all all
crustacean bivalves
s
in 2.14 0.50 Median of pc 1.41 0.10
planktonic
crustacean
s
in 2.14 0.50 Median of all 1.41 0.50
all
invertebrat
es except
bivalves
in 2.14 0.36 Median in,bc 1.74 0.20 Median of bvs 4.29 0.08
insects and all
benthic bivalves
crustacean
s
in 2.14 0.68 Median of pc 1.41 0.20 crayfish cr 1.46 0.08
planktonic
crustacean
s
TL1 1.00 0.60 Median of all 1.41 0.40
all
invertebrat
es except
bivalves
TL1 1.00 0.50 Median of pc 1.41 0.40 Median of in 2.14 0.10
planktonic all insects
1° 1°TL3 1°TL3 1° 1°TL3
TL3 spp used spp TL3 proportio
spp abbrev TT n
F
Fish Median all f+a 1.79 0.4
fish eating
median all
invertebrat
es
Fish Median all f+a 1.79 0.36
fish eating
median all
invertebrat
es
Fish Median all f+a 1.79 0.04
fish eating
median all
invertebrat
es
Effectiv Targe TTFcomposi
e TTF t fish te
TTF
2.03 0.91 1.85
1.93 2.67 5.14
1.78 1.46 2.59
2.11 0.73 1.53
1.93 1.48 2.85
1.16 1.04 1.21
1.28 1.46 1.86
bottom ooze (Becker 1983).
crustacean
Do not distribute, quote or cite
B-138
Draft Document
-------
Referen Site Site Target Fish Fish prey as described in NatureServe Fish prey spp 1° TL2 TTF 1° TL2 1° 1° TL2 2° TL2 2° 2° 2° TL2 3° TL2 3° 3° 3° TL2 4° TL2 4°
ce descript ID fish TTF comment species used spp TL2 proportio spp used TL2 TL2 proportio spp used TL2 TL2 proportio spp used TL2
ion species abbrev TT n spp TT n spp TT n spp
commo . F abrev F abrev F abrev
n name
Default
Default
Default
Default
Default
Default
Default
brook
stickleb
ack
brook
trout
brown
bullhea
d
brown
trout
bullhea
d
channel
catfish
commo
ncarp
1 .69 Eats various aquatic invertebrates
(including eggs and larvae), eggs and
larvae of fishes, and algae. In a Manitoba
lake, was opportunistic but heavily
dependent on arthropods (Moodie 1986).
0.88 Feeds opportunistically on various
invertebrate and vertebrate animals,
including primarily terrestrial and aquatic
insects and planktonic crustaceans.
0.91 Bottom feeder. Young eat chironomid
larvae and small crustaceans. Adults eat
larger insect larvae and fishes, also fish
eggs, mollusks, carrion, and plant
material (Becker 1983, Moyle 1976).
1 .44 Eats aquatic and terrestrial insects and
their larvae, crustaceans (especially
crayfish), molluscs, fishes, and other
animals. In streams, young feed mainly
on aquatic and terrestrial drift
invertebrates; in lakes, they feed on
zooplankton and benthic invertebrates
(Sublette et al. 1990). Large adults feed
on fishes, crayfish, and other benthic
invertebrates.
0 . 82 Black (not exotic to CO and NM):
Omnivorous bottom feeder; often eats
aquatic insects, crustaceans, molluscs,
occasionally fishes and carrion. Stomach
often contain substantial amounts of plant
material of unknown nutritional value
(Moyle 1976). Juveniles planktivorous; at
about 27 mm TL, feed largely on
crustaceans and midge larvae
0.73 Bottom feeder. Young eat mainly small
invertebrates; as they grow, fishes and
crayfish become increasingly important,
though individuals of all sizes eat
abundant aquatic insects. Large fish are
mainly piscivorous (Moyle 1976).
1 .34 Omnivorous; adults eat mainly
invertebrates, detritus, fish eggs, and
plant material (Jester 1974, Becker 1983,
Sublette et al. 1990).
Median of all all 1.41
invertebrates
except
bivalves
Median of all in 2.14
insects
Median of all in 2.14
insects
Median of pc 1.41
planktonic
crustaceans
Median of all in 2.14
insects
Median of all in 2.14
insects
Median of all all 1.41
invertebrates
except
bivalves
0.80 TL1 TL1 1.00 0.20
0.60 crayfish cr 1.46 0.10 Median of bvs 4.29 0.05
all
bivalves
0.68 Median of all 1.41 0.20 Median of bvs 4.29 0.04
all all
invertebrat bivalves
es except
bivalves
0.20 Median of in 2.14 0.12 crayfish cr 1.46 0.08
all insects
0.68 Median of all 1.41 0.20 Median of bvs 4.29 0.04
all all
invertebrat bivalves
es except
bivalves
0.48 Median of pc 1.41 0.20 crayfish cr 1.46 0.08
planktonic
crustacean
s
0.65 TL1 TL1 1.00 0.35
4° 4°TL2 1° 1°TL3 1° TL3 1° 1° TL3
TL2 proportio TL3 spp used spp TL3 proportio
TT n spp abbrev TT n
F . F
Fish Median all f+a 1.79
fish eating
median all
invertebrat
es
Fish Median all f+a 1.79
fish eating
median all
invertebrat
es
Fish Median all f+a 1.79
fish eating
median all
invertebrat
es
Fish Median all f+a 1.79
fish eating
median all
invertebrat
es
Fish Median all f+a 1.79
fish eating
median all
invertebrat
es
0.25
0.08
0.6
0.08
0.24
Effectiv Targe TTFcomposi
e TTF t fish te
TTF
1.33 1.69
2.09 O.S
2.05 0.91
1.73 1.44
2.25
1.85
1.87
2.49
2.05 0.82
1.68
1.86 0.73
1.35
1.27 1.34
1.70
Do not distribute, quote or cite
B-139
Draft Document
-------
Referen Site Site
ce descript ID
ion
Default
Target
fish
species
commo
nname
crappie
Fish Fish prey as described in NatureServe
TTF
2.67 Black: Primarily a midwater feeder;
zooplankton and small Diptera larvae
predominate in the diet of individuals to
12 cm SL, while fishes and aquatic
insects predominate in the diet of larger
Fish prey spp 1° TL2 TTF
comment species used
Median of all
insects
1°TL2
spp
abbrev
in
1°
TL2
TT
F
2.14
1°TL2
proportio
n
0.50
2°TL2
spp used
Median of
planktonic
crustacean
s
2°
TL2
spp
abrev
pc
2°
TL2
TT
F
1.41
2° TL2 3° TL2
proportio spp used
n
0.10
3°
TL2
spp
abrev
3° 3° TL2 4° TL2
TL2 proportio spp used
TT n
F
4°
TL2
spp
abrev
4° 4° TL2
TL2 proportio
TT n
F
1°
TL3
spp
Fish
1°TL3
spp used
Median all
fish eating
median all
invertebrat
es
1°TL3
spp
abbrev
f+a
1°
TL3
TT
F
1.79
1°TL3
proportio
n
0.4
Effectiv Targe TTFcomposi
e TTF t fish te
TTF
Default
Default
Default
Default
Default
Default
Default
1.93 2.67
5.14
creek
chub
cutthroa
t trout
fathead
minnow
flannel
mouth
sucker
flathead
chub
freshwa
ter
drum
gizzard
Do not distribute, quote or cite
individuals. White: eats fishes,
planktonic crustaceans, and aquatic
insects; small individuals eat mostly
zooplankton, fish tend to predominate in
the diet of larger individuals, though
zooplankton also consumed (Moyle
1976)
1.12 Feeds opportunistically on various plants
and animals, from surface drift to
benthos; mostly invertivorous but large
individuals often picivorous (Becker
1983). Chironomid larvae and other
larval insects important in diet of young.
1.07 Opportunistic. Inland cutthroats feed
primarily on insects (aquatic and
terrestrial); often feeds in and especially
downstream from riffle areas; some large
individuals feed mostly on fishes; also
eats zooplankton and crustaceans.
1.57 Feeds opportunistically in soft bottom
mud; eats algae and other plants, insects,
small crustaceans, and other invertebrates
(Becker 1983, Sublette et al. 1990).
1.06 Herbivore, Invertivore Bottom feeder.
Reported to feed on diatoms, algae,
fragments of higher plants, seeds, and
benthic invertebrates (Sigler and Miller
1963; Lee et al. 1980). See Tyus and
Minckley 1988 for possible importance
of Mormon cricket as food source.
1.46 Opportunistic; eats aquatic and terrestrial
insects and some algae (Olund and Cross
1961)
1.48 Young feed mainly on minute
crustaceans; adults mostly are bottom
feeders, eat insect larvae, crustaceans,
fishes, and (mostly in rivers) clams and
snails (Becker 1983, Scott and Grossman
1973, Lee etal. 1980).
1.27 Adults primarily bottom filter-feeding
expected diet
of small
invertebrates
Median of all all
invertebrates
except
bivalves
Median of all
insects
Median of all
insects
Median in,bc
insects and
benthic
crustaceans
Median of all in
insects
Median of all crs
crustaceans
TL1
TL1
1.41
2.14
2.14
1.74
2.14
1.41
1.00
0.70 TL1
1.00
TL1
1.00
0.20
Fish
0.50 Median of crs
all
crustacean
s
0.60 Median of crs
all
crustacean
s
0.75 TL1 TL1
1.41
0.20
Fish
1.41
1.00
0.20 TL1
0.25
TL1 1.00
0.20
0.80 TL1
TL1 1.00
0.44 Median of in 2.14
all insects
0.20
0.40 Median of bvs
all
bivalves
4.29
0.04
Fish
B-140
Median all
fish eating
median all
invertebrat
Median all
fish eating
median all
invertebrat
es
Median all
fish eating
median all
invertebrat
es
f+a
1.79
0.1
1.37
1.12
1.53
f+a
1.79
0.3
1.89
1.07
2.02
1.77
1.55
1.57
1.06
2.77
1.64
f+a
1.79
0.12
1.91
1.86
1.46
1.48
2.79
2.76
1.00
1.27 1.27
Draft Document
-------
Referen Site Site
ce descript ID
ion
Default
Default
Default
Default
Default
Default
Default
Default
Target
fish
species
commo
nname
shad
goldeye
green
sunfish
Iowa
darter
kokane
e
salmon
largemo
uth bass
longnos
edace
longnos
e sucker
mixed
Fish Fish prey as described in NatureServe Fish prey spp
TTF comment
detritivores
1 .27 Young-of-year eat mainly
microcrustaceans, also other
invertebrates. Older individuals eat
mainly aquatic insects obtained at surface
but also various other animals, including
frogs, fishes, and small mammals.
1 .27 Feeds opportunistically on the larger,
more active invertebrates that occur with
them, and on small fishes. Young feed
mostly on crustaceans (zooplankton) and
aquatic insect larvae. Adults eat more
large aquatic and terrestrial insects,
crayfish, and fishes
1 .62 Eats mainly various invertebrates; expected diet
commonly ingested food items of adults of small
are midge larvae, mayfly naiads, and invertebrates
amphipods, and of the young, copepods
and cladocerans. Apparently feeds on
swimming organisms and those on
bottom.
1.19 Zooplankton, insects.
1 .27 Fry feed mainly on zooplankton. Larger
young eat insects, crustaceans, and fish
fry. Adults eat mainly fishes, though
sometimes prefer crayfish or amphibians
(Moyle 1976, Smith 1979).
1 .46 Eats mainly benthic insects, especially
Diptera and mayflies (Becker 1983, Scott
and Grossman 1973); also eats algae and
plant material (Sublette et al. 1990).
Terrestrial insects and fish egs common
in diet of adults from Lake Michigan (see
Sublette et al. 1990).
0.90 Eats mostly bottom invertebrates (Scott
and Grossman 1973).
0.87
1°TL2TTF 1°TL2 1° 1° TL2 2° TL2 2° 2° 2° TL2 3° TL2 3° 3° 3° TL2 4° TL2 4° 4° 4° TL2 1° 1° TL3 1° TL3 1° 1° TL3
species used spp TL2 proportio spp used TL2 TL2 proportio spp used TL2 TL2 proportio spp used TL2 TL2 proportio TL3 spp used spp TL3 proportio
abbrev TT n spp TT n spp TT n spp TT n spp abbrev TT n
F abrev F abrev F abrev F . F
Median in,bc 1.74 1.00
insects and
benthic
crustaceans
Median of all in 2.14 0.58 Median of pc 1.41 0.10 crayfish cr 1.46 0.08 Fish Median all f+a 1.79 0.24
insects planktonic fish eating
crustacean median all
s invertebrat
es
Median of all in 2.14 0.70 amphipods am 1.22 0.16 crayfish cr 1.46 0.08 Median of pc 1.41 0.06
insects planktoni
c
Crustacea
ns
Median of pc 1.41 0.80 Median of in 2.14 0.20
planktonic all insects
crustaceans
Median of all in 2.14 0.10 crayfish cr 1.46 0.10 Fish Median all f+a 1.79 0.8
insects fish eating
median all
invertebrat
es
Median of all in 2.14 0.80 TL1 TL1 1.00 0.20
insects
Median of all all 1.41 1.00
invertebrates
except
bivalves
Median of all all 1.41 1.00
invertebrates
except
bivalves
Effectiv Targe TTFcomposi
e TTF t fish te
TTF
1.74 1.27 2.20
1.93 1.27 2.44
1.90 1.62 3.08
1.56 1.19 1.85
1.79 1.27 2.27
1.91 1.46 2.79
1.41 0.90 1.27
1.41 0.87 1.23
Do not distribute, quote or cite
B-141
Draft Document
-------
Referen Site Site
ce descript ID
ion
Default
Default
Default
Default
Default
Default
Target
fish
species
commo
nname
mosquit
ofish
mottled
sculpin
mountai
n
whitefis
h
norther
npike
norther
n plains
killifish
norther
n
redbelly
dace
Fish Fish prey as described in NatureServe Fish prey spp
TTF comment
0.86 Opportunistic. Inland cutthroats feed
primarily on insects (aquatic and
terrestrial); often feeds in and especially
downstream from riffle areas; some large
individuals feed mostly on fishes; also
eats zooplankton and crustaceans.
1 .38 Benthic feeder; forages among rocks,
mainly on immature aquatic insect larvae,
especially mayflies, chironomid midges,
and stoneflies; larger individuals also eat
caddisflies and crayfish; crustaceans,
annelids, fishes (including fish eggs) and
plant material also may be eaten; may
take swimming prey from water column
(Scott and Grossman 1973, Becker 1983).
1 .38 Feeds actively on aquatic and terrestrial
insects. Also feeds on some fish eggs and
occasionally on fishes. Bottom-oriented
predator (Moyle 1976), occasionally
feeds at surface (Sigler and Sigler 1987).
2.04 Young initially eat large zooplankton and
immature aquatic insects. After 7-10 days
fishes begin to enter diet and eventually
dominate. Adults feed opportunistically
on vertebrates small enough to be
engulfed. (Scotland Grossman 1973).
Sight feeder.
1 .27 Feed effectively at all levels and food Montana field
habits are generalized. Prefer aquatic guide
insects but also feed on plants. (http://fieldgui
de.mt.gov/detai
1_AFCNB0460
O.aspx)
1 .46 Eats mainly diatoms and filamentous
algae, also zooplankton and aquatic
insects.
1°TL2TTF 1°TL2 1° 1° TL2 2° TL2 2° 2° 2° TL2 3° TL2 3° 3° 3° TL2 4° TL2
species used spp TL2 proportio spp used TL2 TL2 proportio spp used TL2 TL2 proportio spp used
abbrev TT n spp TT n spp TT n
F abrev F abrev F
Median of all in 2.14 0.75 Median of crs 1.41 0.25
insects all
crustacean
s
Median of all in 2.14 0.70 Median of crs 1.41 0.10 TL1 TL1 1.00 0.10
insects all
crustacean
s
Median of all in 2.14 0.90
insects
Median in,bc 1.74 0.05
insects and
benthic
crustaceans
Median of all in 2.14 0.80 TL1 TL1 1.00 0.20
insects
TL1 TL1 1.00 0.70 Median of in 2.14 0.15 Median in,bc 1.74 0.15
all insects insects
and
benthic
crustacean
s
4° 4° 4°TL2 1° 1°TL3
TL2 TL2 proportio TL3 spp used
spp TT n spp
abrev F
Fish Median all
fish eating
median all
invertebrat
es
Fish Median all
fish eating
median all
invertebrat
es
Fish Median all
fish eating
median all
invertebrat
es
1° TL3 1° 1° TL3
spp TL3 proportio
abbrev TT n
F
f+a 1.79 0.1
f+a 1.79 0.1
f+a 1.79 0.95
Effectiv Targe TTFcomposi
e TTF t fish te
TTF
1.96 0.86
1.69
1.92 1.38
2.65
2.11
1.79
1.38
2.04
2.90
3.66
1.91
1.27
2.44
1.28
1.46
1.87
Do not distribute, quote or cite
B-142
Draft Document
-------
Referen Site Site
ce descript ID
ion
Default
Default
Default
Default
Default
Default
Default
Default
Target
fish
species
commo
nname
norther
n
squawfi
sh
rainbow
trout
red
shiner
redside
shiner
river
carp sue
ker
roundta
il chub
sacrame
nto
perch
sailfm
molly
Fish Fish prey as described in NatureServe Fish prey spp
TTF comment
1 .46 Small individuals feed primarily on
aquatic and terrestrial insects. Adults feed
on fish, insects, insect larvae, crustaceans
and some plankton during spring and
summer. Fishes are the major component
of the diet in winter.
1.19 In lakes, feeds mostly on bottom-
dwelling invertebrates (e.g., aquatic
insects, amphipods, worms, fish eggs,
sometimes small fish) and plankton. In
streams, feeds primarily on drift
organisms. May ingest aquatic vegetation
(probably for attached invertebrates).
1 .46 Eats various small invertebrates (insects,
crustaceans), plant material (digestibility
may be low), and microorganisms
(Becker 1983). In Virgin River, diet
dominated by Ceratopongidae,
Simuliidae, and Chironomidae (Greger
and Deacon 1988).
1 .46 Feeds mainly on aquatic and terrestrial
insects; also eats molluscs, plankton, and
some small fish and fish eggs. Fry eat
zooplankton and algae.
1 .05 Mostly a bottom feeder, browses on
periphyton associated with submerged
rocks and debris, ingests various small
planktonic plants and animals.
1 .46 Opportunistic; eats available aquatic and
terrestrial insects, gastropods,
crustaceans, fishes, and sometimes
filamentous algae (Sublette et al. 1990).
1 .48 Opportunistic; diet mainly benthic insect
larvae, snails, mid- water insects,
zooplankton, and fishes (Moyle et al.
1989). Young feed mainly on small
crustaceans, but as they grow Sacramento
perch consume more aquatic insects
larvae and pupae. Large adults feed
mainly on other fishes when available.
1 .06 Eats mainly algae, vascular plants,
organic detritus, and mosquito larvae
(and other small invertebrates).
1° TL2 TTF
species used
Median of all
insects
Median of all
insects
Median
insects and
benthic
crustaceans
Median of all
insects
TL1
Median of all
insects
TL1
TL1
1°TL2 1° 1°TL2 2°TL2 2° 2° 2° TL2 3° TL2 3° 3° 3° TL2 4° TL2
spp TL2 proportio spp used TL2 TL2 proportio spp used TL2 TL2 proportio spp used
abbrev TT n spp TT n spp TT n
F abrev F abrev F
in 2.14 0.32 Median of crs 1.41 0.08
all
crustacean
s
in 2.14 0.75
in,bc 1.74 1.00
in 2.14 0.70 Median of pc 1.41 0.10 Median of bvs 4.29 0.10
planktonic all
crustacean bivalves
s
TL1 1.00 0.75 Median of pc 1.41 0.25
planktonic
crustacean
s
in 2.14 0.55 Median of crs 1.41 0.15 Median of bvs 4.29 0.15
all all
crustacean bivalves
s
TL1 1.00 0.75 Median of in 2.14 0.25
all insects
TL1 1.00 0.75 Median of in 2.14 0.25
all insects
4° 4° 4°TL2 1° 1°TL3
TL2 TL2 proportio TL3 spp used
spp TT n spp
abrev F
Fish Median all
fish eating
median all
invertebrat
es
Fish Median all
fish eating
median all
invertebrat
es
Fish Median all
fish eating
median all
invertebrat
es
Fish Median all
fish eating
median all
invertebrat
es
1° TL3 1° 1° TL3
spp TL3 proportio
abbrev TT n
F
f+a 1.79 0.6
f+a 1.79 0.25
f+a 1.79 0.1
f+a 1.79 0.15
Effectiv Targe TTFcomposi
e TTF t fish te
TTF
1.87 1.46
2.73
2.05 1.19
2.44
1.74 1.46
2.53
2.25 1.46
1.10 1.05
2.30 1.46
1.29 1.48
3.28
1.16
3.35
1.90
1.29 1.06
1.36
Do not distribute, quote or cite
B-143
Draft Document
-------
Referen Site Site
ce descript ID
ion
Default
Default
Default
Default
Default
Default
Target
fish
species
commo
nname
sand
shiner
sauger
sculpin
shorthe
ad
redhors
e
smallm
outh
bass
speckle
d dace
Fish Fish prey as described in NatureServe Fish prey spp
TTF comment
1 .83 Eats various aquatic and terrestral
invertebrates (especially chironomids),
algae, and (mainly) bottom particulate
matter (Becker 1983). Winter diet mostly
chironomids larvae and mayfly and
stonefly naiads (Ohio, see Sublette et al.
1990)
1 .82 Larvae eat microcrustaceans. Young eat
zooplankton, immature and adult aquatic
insects, and fish fry; adults eat small
fishes and various invertebrates (Scott
and Grossman 1973), or are almost
exclusively piscivorous (Burkhead and
Jenkins 1991). Sight feeder, adapted to
low light.
1 .29 Benthic feeder; forages among rocks,
mainly on immature aquatic insect larvae,
especially mayflies, chironomid midges,
and stoneflies; larger individuals also eat
caddisflies and crayfish; crustaceans,
annelids, fishes (including fish eggs) and
plant material also may be eaten; may
take swimming prey from water column
(Scott and Grossman 1973, Becker 1983).
1 .05 Invertivore
1 .27 Adults almost entirely piscivorous if
sufficient prey available
1 .46 An omnivorous benthic feeder, at times
feeding on drift in mid- water or rarely at
the surface (Schreiber and Minckley
1981). The diet consists mostly of benthic
insects, also includes other invertebrates,
algae, and detritus (little or no plant
material or detritus in some areas)
(Sublette et al. 1990, Woodbury 1933,
Greger and Deacon 1988). Young feed
mainly on zooplankton.
1°TL2TTF 1°TL2 1° 1° TL2 2° TL2 2° 2° 2° TL2 3° TL2
species used spp TL2 proportio spp used TL2 TL2 proportio spp used
abbrev TT n spp TT n
F abrev F
Median in,bc 1.74 0.75 TL1 TL1 1.00 0.25
insects and
benthic
crustaceans
Median in,bc 1.74 0.36 Median of pc 1.41 0.10
insects and planktonic
benthic crustacean
crustaceans s
Median of all in 2.14 0.70 crayfish cr 1.46 0.15
insects
Median of all all 1.41 1.00
invertebrates
except
bivalves
Median of all in 2.14 0.20
insects
Median of all in 2.14 0.70 Median in,bc 1.74 0.15 TL1
insects insects and
benthic
crustacean
s
3° 3° 3°TL2 4°TL2 4° 4° 4° TL2 1° 1° TL3
TL2 TL2 proportio spp used TL2 TL2 proportio TL3 spp used
spp TT n spp TT n spp
abrev F abrev F
Fish Median all
fish eating
median all
invertebrat
es
Fish Median all
fish eating
median all
invertebrat
es
Fish Median all
fish eating
median all
invertebrat
es
TL1 1.00 0.15
1° TL3 1° 1° TL3
spp TL3 proportio
abbrev TT n
F
f+a 1.79 0.54
f+a 1.79 0.15
f+a 1.79 0.8
Effectiv Targe TTFcomposi
e TTF t fish te
TTF
1.55 1.83
2.84
1.73 1.82
3.16
1.99 1.29
2.57
1.41 1.05
1.86 1.27
1.91 1.46
1.48
2.35
2.78
Do not distribute, quote or cite
B-144
Draft Document
-------
Referen Site Site
ce descript ID
ion
Default
Target
fish
species
commo
nname
stonecat
Fish Fish prey as described in NatureServe Fish prey spp
TTF comment
0.82 Eats mainly bottom invertebrates (insects,
crayfish); sometimes also plant material
and fishes (Becker 1983, Scott and
Grossman 1973).
1° TL2 TTF
species used
Median
insects and
benthic
crustaceans
1°TL2
spp
abbrev
in,bc
1°
TL2
TT
F
1.74
1° TL2 2° TL2
proportio spp used
n
0.70 TL1
2°
TL2
spp
abrev
TL1
2°
TL2
TT
F
1.00
2° TL2 3° TL2
proportio spp used
n
0.20
3°
TL2
spp
abrev
3° 3° TL2 4° TL2
TL2 proportio spp used
TT n
F
4°
TL2
spp
abrev
4° 4° TL2
TL2 proportio
TT n
F
1°
TL3
spp
Fish
1°TL3
spp used
Median all
fish eating
median all
invertebrat
es
1°TL3
spp
abbrev
f+a
1°
TL3
TT
F
1.79
1°TL3
proportio
n
0.1
Effectiv Targe TTFcomposi
e TTF t fish te
TTF
Default
Default
sucker 1.05 White: Larvae feed near surface on
protozoans, diatoms, small crustaceans,
and bloodworms. Adults feed
opportunistically on bottom organisms,
both plant and animal (e.g., chironomid
larvae, zooplankton, small crayfishes)
(Becker 1983, Sublette et al. 1990).
Bluehead: A bottom feeder. Scrapes
algae and other organisms from rocks
with chisel-like ridges inside each lip;
ingests fine organism-laden sediments.
May feed in stream riffles, or deeper
rocky pools; in lakes it may feed over
rocks near shore. May eat aquatic insect
larvae. Flannelmouth: Bottom feeder.
Reported to feed on diatoms, algae,
fragments of higher plants, seeds, and
benthic invertebrates (Sigler and Miller
1963; Lee et al. 1980). See Tyus and
Minckley 1988 for possible importance
of Mormon cricket as food source.
sunfish 2.00 Species present inGEI 2013. Bluegill:
species Feeds opportunistically on aquatic insect
larvae, planktonic crustaceans, flying
insects, snails, and other small
invertebrates; small fishes, fish eggs,
crayfish, and algae sometimes are eaten.
Larvae and juveniles often eat
cladocerans and copepod nauplii. Adults
eats mainly aquatic insects, crayfishes,
and small fishes, or, in some bodies of
water, mostly zooplankton. Feeds at all
levels of water column. Green sunfish:
Feeds opportunistically on the larger,
more active invertebrates that occur with
them, and on small fishes. Young feed
mostly on crustaceans (zooplankton) and
aquatic insect larvae. Adults eat more
large aquatic and terrestrial insects,
crayfish, and fishes. Based on reported
Median of all all 1.41 0.50 TL1
invertebrates
except
bivalves
TL1 1.00 0.50
1.60 0.82
1.20 1.05
1.31
1.27
Median of all in 2.14 0.63 Median of pc
insects planktonic
crustacean
1.41
0.15 crayfish cr
1.46
0.10
Fish
Median all
fish eating
median all
invertebrat
f+a
1.79
0.12
1.92 2.00
3.84
Do not distribute, quote or cite
B-145
Draft Document
-------
Referen
ce
Site
descript
ion
Site
ID
Target
fish
species
commo
nname
Fish
TTF
Fish prey as described in NatureServe Fish prey spp
comment
1° TL2 TTF
species used
1°TL2
spp
abbrev
1°
TL2
TT
F
1°TL2
proportio
n
2°TL2
spp used
2°
TL2
spp
abrev
2°
TL2
TT
F
2°TL2
proportio
n
3°TL2
spp used
3°
TL2
spp
abrev
3°
TL2
TT
F
3°TL2
proportio
n
4°TL2
spp used
4°
TL2
spp
abrev
4°
TL2
TT
F
4°TL2
proportio
n
1°
TL3
spp
1°TL3
spp used
1°TL3
spp
abbrev
1°
TL3
TT
F
1°TL3
proportio
n
Effectiv Targe TTFcomposi
e TTF t fish te
TTF
species in GEI 2013
Default
Default
Default
tilapia 1.48 aureus: Eats mainly phytoplankton.
mossambicus: Nonselective omnivore;
eats planktonic algae, aquatic plants,
invertebrates, and small fishes (Moyle
1976). zilli: Feeds on algae and higher
plants, invertebrates, and occasionally
eats dead or dying fish.
trout 1.19 Rainbow: In lakes, feeds mostly on
species bottom-dwelling invertebrates (e.g.,
aquatic insects, amphipods, worms, fish
eggs, sometimes small fish) and plankton.
In streams, feeds primarily on drift
organisms. May ingest aquatic vegetation
(probably for attached invertebrates).
Brown: Eats aquatic and terrestrial
insects and their larvae, crustaceans
(especially crayfish), molluscs, fishes,
and other animals. In streams, young feed
mainly on aquatic and terrestrial drift
invertebrates; in lakes, they feed on
zooplankton and benthic invertebrates
(Sublette et al. 1990). Large adults feed
on fishes, crayfish, and other benthic
invertebrates.
tui chub 1.46 Adults opportunistic. They feed on plant
material, plankton, insect larvae,
crustaceans, fish fry and fish eggs, etc.
Young feed on zooplankton. Coarse-
rakered form eats more plant material,
fine-rakered form more zooplankton.
Median of all
invertebrates
except
bivalves
Median of all
insects
all
1.41
0.50 TL1
TL1
1.00
0.50
1.20
1.48
1.78
2.14
0.55 crayfish
1.46
0.05
Fish
Median all
fish eating
median all
invertebrat
es
f+a
1.79
0.4
1.97
1.19
2.34
TL1
TL1
1.00
0.40 Median of pc
planktonic
crustacean
1.41
0.28 Median of
all insects
in
2.14
0.28 crayfish
cr
1.46
0.04
1.45
1.46
2.12
Do not distribute, quote or cite
B-146
Draft Document
-------
Referen
ce
Site Site
descript ID
ion
Target Fish
fish TTF
species
commo
nname
Fish prey as described in NatureServe
Fish prey spp
comment
1° TL2 TTF
species used
1°TL2
spp
abbrev
1°
TL2
TT
F
1° TL2 2° TL2
proportio spp used
n
2°
TL2
spp
abrev
2°
TL2
TT
F
2°TL2
proportio
3°TL2
spp used
3°
TL2
spp
abrev
3°
TL2
TT
F
3°TL2
proportio
4°TL2
spp used
4°
TL2
spp
abrev
4°
TL2
TT
F
4°TL2
proportio
1°
TL3
spp
1°TL3
spp used
1° TL3 1° 1° TL3
spp TL3 proportio
abbrev
TT
F
n
Effectiv Targe TTFcomposi
e TTF t fish te
TTF
Default
Default
utah
sucker
walleye
1 .05 Bottom feeder. Varied diet; feeds freely
on both animal and plant organisms, at all
depths throughout the year. Grazes on
filamentous algae.
1 .82 Adults feed opportunistically on various
fishes and larger invertebrates.
Median of all all 1.41 0.50 TL1 TL1 1.00 0.50
invertebrates
except
bivalves
Median in,bc 1.74 0.50
insects and
benthic
crustaceans
Fish Median all f+a 1.79
fish eating
median all
invertebrat
0.5
1.20 1.05
1.76 1.82
1.27
3.21
Default
Default
Default
western 1.25 Opportunistic omnivore; eats mainly
mosquit small invertebrates, often taken near
ofish water surface. Also eats small fishes and,
in the absence of abundant animal food,
algae and diatoms (Moyle 1976).
Mosquito fish are principally carnivorous,
and have strong, conical teeth and short
guts (Meffe et al. 1983, Turner and
Snelson 1984). They are reported to feed
on rotifers, snails, spiders, insect larvae,
crustaceans, algae, and fish fry, including
their own progeny (Barnickol 1941,
Minckley 1973, Meffe and Crump 1987).
Cannibalism has been documented by
several authors (Seale 1917, Krumholz
1948, Walters and Legner 1980,
Harrington and Harrington 1982). Plant
material is taken occasionally (Bamickol
1941) and may make up a significant
portion of the diet during periods of
scarcity of animal prey (Harrington and
Harrington 1982). Grubb (1972) showed
that anuran eggs from temporary ponds
were preferentially selected over those
breeding in permanent systems.
westslo 1.20 Opportunistic. Inland cutthroats feed
pe primarily on insects (aquatic and
cutthroa terrestrial); often feeds in and especially
t trout downstream from riffle areas; some large
individuals feed mostly on fishes; also
eats zooplankton and crustaceans.
white 1.48 Eats fishes, zooplankton, aquatic insects,
bass oligochaetes, and crayfish; fishes often
dominate diet of adults; diet may vary
from place to place (Moyle 1976,
Do not distribute, quote or cite
Median of all
insects
2.14
0.75 Median of
all
crustacean
s
1.41
0.25
1.96 1.25
2.46
Median of all
insects
Median of all
insects
in
2.14
in
2.14
0.45 Median of pc
planktonic
crustacean
0.30 Median of pc
planktonic
crustacean
s
B-147
1.41
0.10
1.41
0.05 crayfish cr
1.46
0.05
Fish Median all f+a 1.79
fish eating
median all
invertebrat
Fish Median all f+a 1.79
fish eating
median all
invertebrat
0.45
1.91 1.20
2.29
1.86 1.48
2.76
Draft Document
-------
Referen Site Site
ce descript ID
ion
Default
Default
Default
Default
Default
Saikiet
al. 1993
Saikiet
al. 1993
Saikiet
al. 1993
Target
fish
species
commo
nname
white
crappie
white
sturgeo
n
white
sucker
wiper
yellow
perch
bluegill
largemo
uth bass
western
mosquit
ofish
Fish Fish prey as described in NatureServe
TTF
Sublette et al. 1990).
2.67 Eats fishes, planktonic crustaceans, and
aquatic insects; small individuals eat
mostly zooplankton, fish tend to
predominate in the diet of larger
individuals, though zooplankton also
consumed (Moyle 1976)
1 .27 A bottom feeder. Young feed mostly on
the larvae of aquatic insects, crustaceans,
and molluscs. A significant portion of the
diet of larger sturgeon consists offish.
1.18 Adults feed opportunistically on bottom
organisms, both plant and animal (e.g.,
chironomid larvae, zooplankton, small
crayfishes) (Becker 1983, Sublette et al.
1990).
1 .48 adults are predatory on fishes and larger
crustaceans (Hassler 1988).
1 .42 Larvae and young primarily zooplankton
feeders; older young eat mostly
invertebrates associated with bottom and
with aquatic plants; adults feed among
plants and along bottom on larger
invertebrates and small fishes (Moyle
1976).
1.48 site- specific: 0.23 chironomid; 0.3
microcrustacea; 0.47 amphipod
1.27 site- specific: 0.72 fish; 0.28 crayfish
1.25 site- specific: 0.89 molluscs, and insects;
0.065 chironomid; 0.045 microcrustacea
Fish prey spp
comment
expected
common spp in
benthos
stomach
analysis
stomach
analysis
stomach
contents show
a large
terrestrial
component
1° TL2 TTF
species used
Median of all
insects
Median
insects and
benthic
crustaceans
TL1
crayfish
Median
insects and
benthic
crustaceans
amphipods
crayfish
Median
insects and
benthic
crustaceans
1°TL2 1° 1°TL2 2°TL2 2° 2° 2° TL2 3° TL2 3° 3° 3° TL2 4° TL2
spp TL2 proportio spp used TL2 TL2 proportio spp used TL2 TL2 proportio spp used
abbrev TT n spp TT n spp TT n
F abrev F abrev F
in 2.14 0.50 Median of pc 1.41 0.10
planktonic
crustacean
s
in,bc 1.74 0.31 Median of bvs 4.29 0.09
all bivalves
TL1 1.00 0.50 Median of in 2.14 0.30 Median of pc 1.41 0.10 crayfish
all insects planktonic
crustacean
s
cr 1.46 0.20
in,bc 1.74 0.64 Median of pc 1.41 0.13 TL1 TL1 1.00 0.07
planktonic
crustacean
s
am 1.22 0.47 Median of pc 1.41 0.30 midges mg 1.90 0.23
planktonic
crustacean
s
cr 1.46 0.27 creek chub CrC 1.12 0.73
in,bc 1.74 0.89 midges mg 1.90 0.07 Median of pc 1.41 0.05
planktonic
crustacean
s
4° 4° 4°TL2 1° 1°TL3
TL2 TL2 proportio TL3 spp used
spp TT n spp
abrev F
es
Fish Median all
fish eating
median all
invertebrat
es
Fish Median all
fish eating
median all
invertebrat
es
cr 1.46 0.10
Fish Median all
fish eating
median all
invertebrat
es
Fish Median all
fish eating
median all
invertebrat
es
1° TL3 1° 1° TL3
spp TL3 proportio
abbrev TT n
F
f+a 1.79 0.4
f+a 1.79 0.6
f+a 1.79 0.8
f+a 1.79 0.16
1.93 2.67
2.00 1.27
1.43 1.18
1.72 1.48
1.65 1.42
1.43 1.48
1.21 1.27
1.74 1.25
5.14
2.53
2.55
2.35
2.12
1.54
2.18
Do not distribute, quote or cite
B-148
Draft Document
-------
Referen
ce
Formatio
n2012
Fonnatio
n2012
Fonnatio
n2012
Fonnatio
n2012
Fonnatio
n2012
Fonnatio
n2012
Fonnatio
n2012
Fonnatio
n2012
Fonnatio
n2012
Fonnatio
n2012
Fonnatio
n2012
Fonnatio
n2012
Fonnatio
n2012
Fonnatio
n2012
Site
descript
ion
Crow
Creek -
CC150
Crow
Creek -
CC150
Crow
Creek -
1A
Crow
Creek -
1A
Crow
Creek -
CC350
Crow
Creek -
CC350
Crow
Creek -
3A
Crow
Creek -
3A
Crow
Creek -
CC75
Crow
Creek -
CC75
Deer
Creek
Deer
Creek
Hoopes
Spring -
HS
Hoopes
Spring -
HS
Site
ID
CC-
150
CC-
150
cc-
1A
cc-
1A
CC-
350
cc-
350
cc-
3A
CC-
3A
CC-
75
cc-
75
DC-
600
DC-
600
HS
HS
Target
fish
species
commo
nname
brown
trout
sculpin
brown
trout
sculpin
brown
trout
sculpin
brown
trout
sculpin
brown
trout
sculpin
brown
trout
sculpin
brown
trout
sculpin
Fish
TTF
1.44
1.29
1.44
1.29
1.44
1.29
1.44
1.29
1.44
1.29
1.44
1.29
1.44
1.29
Fish prey as described in NatureServe Fish prey spp 1° TL2 TTF 1° TL2 1° 1° TL2 2° TL2 2° 2° 2° TL2 3° TL2 3° 3° 3° TL2 4° TL2 4° 4° 4° TL2 1°
comment species used spp TL2 proportio spp used TL2 TL2 proportio spp used TL2 TL2 proportio spp used TL2 TL2 proportio TL3
abbrev TT n spp TT n spp TT n spp TT n spp
F abrev F abrev F abrev F
Proportions as described in table C-2.
Proportions as described in table C-2.
Proportions as described in table C-2.
Proportions as described in table C-2.
Proportions as described in table C-2.
Proportions as described in table C-2.
Proportions as described in table C-2.
Proportions as described in table C-2.
Proportions as described in table C-2.
Proportions as described in table C-2.
Proportions as described in table C-2.
Proportions as described in table C-2.
Proportions as described in table C-2.
Proportions as described in table C-2.
Median of all in 2.14
insects
Median of all in 2.14
insects
Median of all in 2.14
insects
Median of all in 2.14
insects
Median of all in 2.14
insects
Median of all in 2.14
insects
Median of all in 2.14
insects
Median of all in 2.14
insects
Median of all in 2.14
insects
Median of all in 2.14
insects
Median of all in 2.14
insects
Median of all in 2.14
insects
Median of all in 2.14
insects
Median of all in 2.14
insects
0.73 midges mg 1.90
0.73 midges mg 1.90
0.89 midges mg 1.90
0.89 midges mg 1.90
0.93 midges mg 1.90
0.93 midges mg 1.90
0.91 blackworm bw 1.29
s
0.91 blackworm bw 1.29
s
0.61 midges mg 1.90
0.61 midges mg 1.90
0.77 midges mg 1.90
0.77 midges mg 1.90
0.50 Median of bvs 4.29
all bivalves
0.50 Median of bvs 4.29
all bivalves
0.27
0.27
0.09 Median of bvs 4.29 0.02
all
bivalves
0.09 Median of bvs 4.29 0.02
all
bivalves
0.07
0.07
0.05 Median of bvs 4.29 0.04
all
bivalves
0.05 Median of bvs 4.29 0.04
all
bivalves
0.37 blackwor bw 1.29 0.02
ms
0.37 blackwor bw 1.29 0.02
ms
0.21 blackwor bw 1.29 0.02
ms
0.21 blackwor bw 1.29 0.02
ms
0.33 blackwor bw 1.29 0.18
ms
0.33 blackwor bw 1.29 0.18
ms
1°TL3 1°TL3 1° 1°TL3 Effectiv Targe TTFcomposi
spp used spp TL3 proportio e TTF t fish te
abbrev TT n TTF
F
2.08 1.44 3.00
2.08 1.29 2.69
2.16 1.44 3.12
2.16 1.29 2.80
2.13 1.44 3.07
2.13 1.29 2.75
2.19 1.44 3.15
2.19 1.29 2.82
2.04 1.44 2.94
2.04 1.29 2.63
2.08 1.44 2.99
2.08 1.29 2.68
2.72 1.44 3.92
2.72 1.29 3.51
Do not distribute, quote or cite
B-149
Draft Document
-------
Referen
ce
Formatio
n2012
Fonnatio
n2012
Fonnatio
n2012
Fonnatio
n2012
Fonnatio
n2012
Fonnatio
n2012
Fonnatio
n2012
Fonnatio
n2012
Site
descript
ion
Hoopes
Spring -
HS3
Hoopes
Spring -
HS3
Sage
Creek -
LSV2C
Sage
Creek -
LSV2C
Sage
Creek -
LSV4
Sage
Creek -
LSV4
South
Fork
Tincup
Cr.
South
Fork
Tincup
Cr.
Site
ID
HS-3
HS-3
LSV-
2C
LSV-
2C
LSV-
4
LSV-
4
SFTC
-1
SFTC
-1
Target
fish
species
commo
nname
brown
trout
sculpin
brown
trout
sculpin
brown
trout
sculpin
brown
trout
sculpin
Fish Fish prey as described in NatureServe
TTF
1 .44 Proportions as described in table C-2.
1 .29 Proportions as described in table C-2.
1 .44 Proportions as described in table C-2.
1 .29 Proportions as described in table C-2.
1 .44 Proportions as described in table C-2.
1 .29 Proportions as described in table C-2.
1 .44 Proportions as described in table C-2.
1 .29 Proportions as described in table C-2.
Fish prey spp 1° TL2 TTF
comment species used
Median of all
insects
Median of all
insects
Median of all
insects
Median of all
insects
Median of all
insects
Median of all
insects
Median of all
insects
Median of all
insects
1° TL2 1°
spp TL2
abbrev TT
F
in 2.14
in 2.14
in 2.14
in 2.14
in 2.14
in 2.14
in 2.14
in 2.14
1° TL2 2° TL2 2°
proportio spp used TL2
n spp
abrev
0.55 Median of crs
all
crustacean
s
0.55 Median of crs
all
crustacean
s
0.91 Median of crs
all
crustacean
s
0.91 Median of crs
all
crustacean
s
0.65 midges mg
0.65 midges mg
0.97 Median of bvs
all bivalves
0.97 Median of bvs
all bivalves
2°
TL2
TT
F
1.41
1.41
1.41
1.41
1.90
1.90
4.29
4.29
2°TL2 3°TL2 3° 3° 3° TL2 4° TL2 4° 4° 4° TL2 1° 1° TL3 1° TL3 1° 1° TL3
proportio spp used TL2 TL2 proportio spp used TL2 TL2 proportio TL3 spp used spp TL3 proportio
n spp TT n spp TT n spp abbrev TT n
abrev F abrev F . F
0.37 midges mg 1.90 0.08
0.37 midges mg 1.90 0.08
0.05 Median of bvs 4.29 0.04
all
bivalves
0.05 Median of bvs 4.29 0.04
all
bivalves
0.34 blackwor bw 1.29 0.02
ms
0.34 blackwor bw 1.29 0.02
ms
0.03
0.03
Effectiv Targe
e TTF t fish
TTF
1.85 1.44
1.85 1.29
2.19 1.44
2.19 1.29
2.07 1.44
2.07 1.29
2.21 1.44
2.21 1.29
TTFcomposi
te
2.67
2.39
3.16
2.83
2.98
2.67
3.18
2.85
Do not distribute, quote or cite
B-150
Draft Document
-------
Table B-9. Calculation of site-specific invertebrate proportions using invertebrate counts in Formation 2012
Order
Ephemeropt
era
Ephemeropt
era
Ephemeropt
era
Ephemeropt
era
Ephemeropt
era
Ephemeropt
era
Ephemeropt
era
Ephemeropt
era
Ephemeropt
era
Ephemeropt
era
Ephemeropt
era
Ephemeropt
era
Plecoptera
Plecoptera
Plecoptera
Plecoptera
Plecoptera
Plecoptera
Trichoptera
Trichoptera
Trichoptera
Trichoptera
Genus
Atenella
margarita
Baetis spp.
Centroptilum
conturbatum
Cinygmula
spp.
Diphetor
hageni
Drunella
coloradensis
Drunella
grandis
Epeorus
longimanus
Ephemerella
dorothea
infrequens
Ephemerella
aurivillii
Paraleptophle
bia spp.
Tricorythodes
minutus
Hesperoperla
pacifica
Isoperla sp.
Malenka sp.
Pteronarcys
sp.
Skwala sp.
Sweltsa sp. p
Agapetus sp.
Arctopsyche
sp.
Brachycentrus
sp.
Cheumatopsy
Do not distribute, quote
Habitat
/
Behavl
or
CN
sw
sw
CN
SW
CN
CN
CN
CN
CN
SW
CN
CN
CN
CN/SP
CN/SP
CN
CN
CN
CN
CN
CN
or cite
Function
al
Feeding
Groups
CG
CG
CG
sc
CG
P
P
SC
CG
CG
CG
CG
P
P
SH
SH
P
P
SC
P
F
F
Toleran Stream SF Tincup Creek Crow Creek Deer Creek
ce
Locatio SFTC1 CC75 CC150 CC350 DC600
n
Date 8/29/2007 9/9/200 9/2/200 8/23/200 9/3/2008 9/1/200 8/24/200 9/3/200 9/1/200 8/23/200 9/4/200 9/7/200 8/27/200 9/8/2008
867 67867867
3 2
5 3 5 56 14 85 89 27 90 68 38 61 253 76 67
2
4 2 14
5 1
0 1 771
0 24 93 3 47
0 453
153 1
15 7
1 2 12 3 9 11 4 11 11
4 2
1 21 12 3 11 21 62 13
2 7 11 5 7
2 10 33 5 25 16 5 14 3 2 4 14 30
0 21
2 3 4 14 1 18
17 13 13 35
0
1 4 18 9 35 2 14 6
1 4 4 29 3 88 4 13 3
B-151
Hoopes Spring Sage Creek Crow Creek
HS HS3 LSV2C LSV4 CC1A CC3A Tota
1
9/8/200 8/24/2007 9/4/200 9/6/2006 8/28/20 9/5/2008 9/8/200 8/28/20 9/5/200 9/5/200 9/1/200 8/25/20 9/6/200 9/4/200 8/26/20 9/7/200
6 8 07 6 07 8 6 6 07 8 6 07 8
3 5
76 9 2 56 249 7 316 27 53 46 57 32 62 56 61 2°41
1 1
16
11 963 3°
79
13 53 1
14 j 38
12
25
51522 1
12
57 4 2 2 1 75
8 7 5 3 25
20 23 94 3 15 21?
1 32613 46
21 4 9 3 29 1 2 212
3 24
2 4 37
14 2 2 77
2 2
13 33 34 23 191
3 17 65 6 153 29 4 20 73 11 27 61 18 635
8 13 21
Draft Document
-------
Order
Trichoptera
Trichoptera
Trichoptera
Trichoptera
Trichoptera
Trichoptera
Trichoptera
Trichoptera
Trichoptera
Trichoptera
Trichoptera
Trichoptera
Trichoptera
Trichoptera
Trichoptera
Trichoptera
Trichoptera
Coleoptera
Coleoptera
Coleoptera
Coleoptera
Coleoptera
Coleoptera
Coleoptera
Coleoptera
Genus
che sp.
Dicosmoecus
sp.
Dolophilodes
sp.
Glossoma sp.
Helicopsyche
sp.
Hesperophyla
xsp.
Hydropsyche
sp.
Hydroptila sp.
Lepidostoma
spp.
Micrasema sp.
Neothremma
sp.
Oecetis
disjuncta
Onocosmoecu
ssp.
Oligophlebod
es sp.
Parapsyche
sp.
Psychoglypha
sp.
Rhyacophila
spp.
Wormaldia
spp.
Ametor sp.
Brychius sp.
Cleptelmis sp.
Dubiraphia sp.
Heterlimnius
corpulentus
Optioservus
quadrimaculat
us
Oreodytes sp.
Paracymus sp.
Habitat
/
Behavl
or
BU
CN
CN
CN
CN
CN
CN
SP/CB
CN
CN
CN
CB
CN
CN
SP/CB
CN
CN
sw
CB
CN
CN
CN/BU
CN
SW/DV
CN
Function Toleran Stream SF Tincup Creek Crow Creek Deer Creek Hoopes Spring Sage Creek Crow Creek
al ce
Feeding
Locatio SFTC1 CC75 CC150 CC350 DC600 HS HS3 LSV2C LSV4 CC1A CC3A Tota
n 1
Groups Date 8/29/2007 9/9/200 9/2/200 8/23/200 9/3/2008 9/1/200 8/24/200 9/3/200 9/1/200 8/23/200 9/4/200 9/7/200 8/27/200 9/8/2008 9/8/200 8/24/2007 9/4/200 9/6/2006 8/28/20 9/5/2008 9/8/200 8/28/20 9/5/200 9/5/200 9/1/200 8/25/20 9/6/200 9/4/200 8/26/20 9/7/200
867 67867867 6 8 07 6 07 866 07 86 07 8
SH
F
SC
SC
SH
F
SC
SH
SH
SC
P
SH
SC
P
CG
P
F
P
SC
CG/SC
CG/SC
CG/SC
CG/SC
P
P/OM
1 3 2
1 25 3
0 4
3 3 5 4 5 93
5
4 5 47 50 23 29 17 11 74 97 41 1 14 2
6 8911
1 13 7 68 67 6 16
1 8 9 65 1 18 14 3 28
0 4
82 23
1 1
1 11
1 16 5 7
1 3
0 7345 5 9 17 16 3 5 9 16 23 83
3 3 18 15 2567
5
72 13
4 3 26 4
4 3
4 30 32
4 97 267 43 109 68 40 205 153 78 162 167 7 2 5 12
5 6 1
5 1
2 7
28
4
22 19 81 214
3 1 48 14 4 7°
2 8 9 11 53 63 91 29 105 79 151 WU
1 16 1 2 39
2 13 4 2 6 141
5 4 76 3 3 36 273
4
3 7 1?
1
2 13
121 32
3
11 13 7 236
39 1 3 1 2 11 7?
1 1
10 16
5 1 1 6 46
3
5 6?
2556
5 21 33 18 132 151 27 153 74 246 69 83 129
7
1
Do not distribute, quote or cite
B-152
Draft Document
-------
Order
Coleoptera
Megaloptera
Odonata
Hemiptera
Diptera
Diptera
Diptera
Diptera
Diptera
Diptera
Diptera
Diptera
Diptera
Diptera
Diptera
Diptera
Diptera
Diptera
Diptera
Diptera
Chironomid
ae (family)
Hiradinea
(class)
Collembola
Oligochaeta
(class)
Bivalvia
(class)
Gastropoda
(class)
Gastropoda
(class)
Gastropoda
(class)
Gastropoda
Genus
Zaitzevia
paravula
Sialis sp.
Ophiogomphu
ssp.
Sigara sp.
Anopheles sp.
Antocha sp.
Atherix sp.
Chelifera sp.
Dixa sp.
Empididae
Ephydridae
Glutops sp.
Hexatoma
Limnophila
sp.
Muscidae
Pericoma sp.
Probezzia sp.
Ptychoptera
sp.
Simulium sp.
Tipula sp.
Chironomidae
Helobdella sp.
Collembola
Oligochaeta
Pisidium sp.
Fossaria sp.
Amnicola sp.
Gyraulus sp.
Mentus sp.
Do not distribute, quote
Habitat
/
Behavl
or
CN/BU
BU/CB
BU
SW
SW
BU
BU
SP/BU
BU
SP/BU
BU
BU
BU
BU
BU
BU
CN
BU
BU/SP
BU
CN
CN
CN
CN
or cite
Function Toleran Stream SF Tincup Creek Crow Creek Deer Creek Hoopes Spring Sage Creek Crow Creek
al ce
Feeding
Locatio SFTC1 CC75 CC150 CC350 DC600 HS HS3 LSV2C LSV4 CC1A CC3A Tota
n 1
Groups Date 8/29/2007 9/9/200 9/2/200 8/23/200 9/3/2008 9/1/200 8/24/200 9/3/200 9/1/200 8/23/200 9/4/200 9/7/200 8/27/200 9/8/2008 9/8/200 8/24/2007 9/4/200 9/6/2006 8/28/20 9/5/2008 9/8/200 8/28/20 9/5/200 9/5/200 9/1/200 8/25/20 9/6/200 9/4/200 8/26/20 9/7/200
CG/SC
P
P
P
F
CG
P
CG
CG
P
CG
P
P
P
P
P
CG
F
SH
CG/SH/P
PA/P
CG
F
SC
SC
SC
SC
4
4
1
10
8
3
2
6
1
6
6
3
2
4
6
6
7
6
4
6
g
5
8
8
5
867 67867867 6 8 07 6 07 866 07 86 07 8
170 57 5 4 1 1 7 23 5 18 33 2 2 7 2 6 16 11 8 2 13 366
1131 6
2 7 9
5 5
1 1
5146 18 2 1 37
26 22 24 3 44 H9
21 74 1 5 20
13 13
152 8
111 3
121 4
19 91 541 9 4 5 16 1 74
133 5 5 3 9 29
1 3 4
2 1 3
3 211 2 2 11
1 !
18 78 5 30 26 49 17 17 5 102 9 15 8 54 13 21 38 24 25 24 12 114 35 8 1 26 31 760
73 1 3 3 2233 27
188 195 173 143 99 143 68 10 30 33 88 151 92 124 25 23 83 20 43 91 149 36 56 35 41 21 8 2168
1 1
2 2
5 15 7 2 6 4 7 8 3 5 3 5 19 72 101 5 3 34 989 19 56 4°5
224 2 262522 12 311 23 69
2 1 2 52 57 27 4 4 8 15 1 1 174
2211 31
1 1
6 6
B-153 Draft Document
-------
Order Genus Habitat Function
/ al
Behavi Feeding
or
Groups
(class)
Gastropoda Physella sp. CN SC
(class)
Gastropoda Valvata sp. CN SC
(class)
Amphipoda Gammarus sp. SW/BU OM
Ostracoda Ostracoda SW CG
Tricladida Polycelis OM
coronata
Acari Acari P
(subclass)
Toleran Stream
ce
Locatio
n
Date
8
6
8
1
8
% Subsampled
Total abundance
Total
taxa
Total Counts
Density (#/lm2)
SF Tincup Creek Crow Creek
SFTC1 CC75 CC150 CC350
8/29/2007 9/9/200 9/2/200 8/23/200 9/3/2008 9/1/200 8/24/200 9/3/200 9/1/200 8/23/200
867 67867
19 3 2 1 3
1
4
2 234 26
50 50 12.5 12.5 66.6 12.5 25 50 25 12.5
394 486 516 506 494 465 482 534 477 536
24 19 27 25 22 26 24 16 23 24
788 972 4128 4048 741.7417 3720 1928 1068 1908 4288
2835 3496 14849 14561 2668 13381 6935 3842 6863 15424
394 486 516 506 494 465 482 534 477 536
880 1516 1481
Deer Creek Hoopes Spring
DC600 HS HS3
9/4/200 9/7/200 8/27/200
867
1
7
50 100 33.3
492 420 478
21 23 23
984 420 1435-43_
3540 1511 5163
492 420 478
1505
9/8/2008 9/8/200 8/24/2007 9/4/200
6 8
114 55 7
1
2
75 50 33.3 100
409 498 415 91
16 15 13 14
545.3333 996 1246.246 91
1962 3583 4483 327
409 498 415 91
1307 1004
9/6/2006 8/28/20 9/5/2008
07
2 6
2 4 13
460 2 9
12.5 33.3 100
596 470 280
21 22 14
4768 141L4j 280
17151 5077 1007
596 470 280
1346
9/8/200
6
14
8
30
25
541
23
2164
7784
541
Sage Creek
LSV2C
8/28/20 9/5/200
07 8
32
1 12
13 8
25 75
532 445
21 17
2128 593'33
7655 2134
532 445
1518
LSV4
9/5/200
6
1
1
12.5
445
21
3560
12806
445
445
Crow Creek
CC1A CC3A Tota
9/1/200 8/25/20 9/6/200
6 07 8
2
2
1
2 2
25 25 50
487 452 463
20 18 22
1948 1808 926
7007 6504 3331
487 452 463
1402
1
9/4/200 8/26/20 9/7/200
6 07 8
3 23 288
1
44
525
4
5 35
25 25 50 j,387
465 503 500
30 20 15
1860 2012 1000
6691 7237 3597
465 503 500
1468 i387
Functional Feeding Groups (FFG): CG = Collector-Gatherer, SC = Scraper, F = Filterer, P = Predator, SH = Shredder, OM = Omnivore Habitat/Behavior (Hab/Beh): BU = Burrower, SW = Swimmer, CN = Clinger, CB = Climber, SP =
Sprawler, DV = Diver
Do not distribute, quote or cite
B-154
Draft Document
-------
Table B-10. Summary of Formation 2012 Invertebrate Data.
Phylum Subphylum Class Subclass Infraclas Superorder Order Lookup ID Common SFTC1 CC75 CC150 CC350 DC600 HS HS3 LSV2C LSV4 CC1A CC3A
s name
Count Proportio Count Proportio Count Proportio Count Proportio Count Proportio Count Proportio Count Proportio Count Proportio Count Proportio Count Proportio Count Proportio
nnnnnn n nnnn
Arthropoda Insecta Pterygota Ephemeropteroide Ephemeropter Ephemeropter Mayflies
a a a
Arthropoda Insecta Pterygota Exopterygota Plecoptera Plecoptera Stoneflie
s
Arthropoda Insecta Amphiesmenopter Trichoptera Trichoptera Caddisflies
a
Arthropoda Insecta Pterygota Neoptera Endopterygota Coleoptera Coleoptera Beetles
Arthropoda Insecta Neoptera Megaloptera Megaloptera Alderflies,
dobsonflies
and fishflies
Arthropoda Insecta Pterygota Odonatoptera Odonata Odonata Dragonflies
and
damselflies
Arthropoda Insecta Neoptera Paraneoptera Hemiptera Hemiptera True bugs
(cicadas,
aphids,
planthoppers,
leafhoppers,
shield bugs)
Arthropoda Insecta Panorpida Diptera Diptera True flies
Arthropoda Insecta Chironomidae Chironomidae Midges
(family) (family)
Annelida Clitellata Hirudine Hirudinea Leeches
a (class)
Arthropoda Entognatha Collembola Collembola Springtails
(not insects!)
Annelida Clitellata Oligochaeta Oligochaeta Worms
(class)
Mollusca Bivalvia Bivalvia Clams
(class)
Mollusca Gastropoda Gastropoda Snails and
(class) slugs
Arthropoda Crustacea Malacostraca Amphipoda Amphipoda Crustaceans
Arthropoda Crustacea Ostracoda Ostracoda Sea shrimp
Platyhelminthes Turbellari Tricladida Tricladida Flatworms
a
Arthropoda Chelicerata Arachnida Acari Acari Mites and
(subclass) ticks
Total
36 0.04
107 0.12
30 0.03
631 0.72
1 0.00
5 0.01
42 0.05
5 0.01
2 0.00
21 0.02
880
185 0.12
85 0.06
268 0.18
234 0.15
4 0.00
143 0.09
556 0.37
24 0.02
6 0.00
7 0.00
4 0.00
1516
231 0.16
40 0.03
283 0.19
408 0.28
1 0.00
103 0.07
385 0.26
1 0.00
2 0.00
17 0.01
2 0.00
1 0.00
7 0.00
1481
192 0.13
18 0.01
539 0.36
457 0.30
145 0.10
108 0.07
16 0.01
11 0.01
4 0.00
15 0.01
1505
444 0.34
195 0.15
229 0.18
76 0.06
55 0.04
272 0.21
27 0.02
8 0.01
1 0.00
1307
115 0.11
59 0.06
34 0.03
18 0.02
29 0.03
241 0.24
178 0.18
9 0.01
319 0.32
2 0.00
1004
325 0.24
20 0.01
230 0.17
65 0.05
89 0.07
106 0.08
3 0.00
2 0.00
16 0.01
19 0.01
471 0.35
1346
421 0.28
26 0.02
324 0.21
327 0.22
85 0.06
154 0.10
43 0.03
12 0.01
54 0.04
21 0.01
51 0.03
1518
56 0.13
30 0.07
135 0.30
29 0.07
36 0.08
149 0.33
8 0.02
1 0.00
1 0.00
445
168 0.12
11 0.01
325 0.23
507 0.36
2 0.00
221 0.16
127 0.09
9 0.01
4 0.00
21 0.01
2 0.00
1 0.00
4 0.00
1402
136 0.09
22 0.01
623 0.42
311 0.21
7 0.00
166 0.11
70 0.05
75 0.05
24 0.02
29 0.02
5 0.00
1468
Midge 0.00 0.37 0.26 0.07 0.21 0.24 0.08 0.10 0.33 0.09 0.05
Other insects 0.97 0.61 0.72 0.90 0.76 0.25 0.54 0.78 0.64 0.88 0.86
Molluscs 0.03 0.01 0.00 0.01 0.01 0.33 0.01 0.04 0.00 0.02 0.04
Do not distribute, quote or cite
B-155
Draft Document
-------
Phylum Subphylum Class Subclass Infraclas Superorder Order
s
Lookup ID Common
name
Crustaceans
Annelids
Other
SFTC1
Count Proportio
n
0.00
0.01
0.00
CC75
Count Proportio
n
0.00
0.02
0.00
CC150
Count Proportio
n
0.00
0.01
0.01
CC350 DC600
Count Proportio Count
n
0.00
0.01
0.01
HS
Proportio Count
n
0.00
0.02
0.00
HS3
Proportio Count
n
0.00
0.18
0.00
LSV2C
Proportio Count
n
0.36
0.00
0.00
Proportio
n
0.05
0.03
0.00
LSV4 CC1A
Count Proportio Count
n
0.00
0.02
0.00
CC3A
Proportio Count
n
0.00
0.01
0.00
Proportio
n
0.00
0.05
0.00
Total
1.00000
1.00000
1.00000
1.00000
1.00000
1.00000
1.00000
1.00000
1.00000
1.00000
1.00000
Take the top 3
that are above
1%
Insects 0.97
Mollusc 0.03
s
Insects 0.61
Midge 0.37
Worm 0.02
s and
leeches
Insect 0.73
s
Midge 0.27
Insects 0.93
Midge 0.07
Insects 0.77
Midge 0.21
Worms 0.02
and
leeches
Insects 0.50
Mollusc 0.33
s
Worms 0.18
and
leeches
Insects
0.55
Insects
0.91
Crustacean 0.37
s
Midge
0.08
Crustacean 0.05
s
Molluscs 0.04
Insects 0.65
Midge 0.34
Worm 0.02
s and
leeches
Insects 0.89
Midge 0.09
Mollusc 0.02
s
Insects
0.91
0.05
Worms
and
leeches
Mollusc 0.04
Do not distribute, quote or cite
B-156
Draft Document
-------
APPENDIX C: SUMMARIES OF CHRONIC STUDIES
CONSIDERED FOR CRITERIA DERIVATION
White sturgeon C-2
Sacramento splittail C-l 2
Fathead minnow C-14
Flannelmouth & razorback suckers C-20
Northern pike C-23
Chinook salmon C-25
Rainbow trout & brook trout C-30
Cutthroat trout C-50
Dolly varden C-67
Brown trout C-70
Desert pupfish C-92
Eastern and western mosquitofish C-109
Striped bass C-l 11
BluegillsunfishC-112
Largemouth bass C-l60
See Appendix E for descriptions of other, less conclusive studies with:
Rainbow trout
Fathead minnow
Sacramento splittail
White sucker
See Appendix E for descriptions of invertebrate studies.
Do not distribute, quote or cite C-l Draft Document
-------
Tashjian, D.H., S.J. The, A. Sogomoyan and S.S.O. Hung. 2006. Bioaccumulation and chronic toxicity
of dietary L-selenomethionine in juvenile white sturgeon (Acipenser transmontanus}. Aquatic
Toxicol.79:401-409.
Test Organism:
Exposure Route:
Test Duration:
Study Design:
Effects Data:
White sturgeon (Acipenser transmontanus)
Dietary only
Seleno-L-methionine was added to an artificial diet consisting of vitamin-free
casein, wheat gluten, egg albumin, dextrin, vitamin mix, BTM-mineral mix,
; O ? OO 5 ; ; 5
cellulose, corn oil, cod liver oil, choline chloride and santoquin; the measured
dietary concentrations were 0.4, 9.6, 20.5, 41.7, 89.8, 191.1 mg Se/kg dw.
8 weeks
25 juvenile white sturgeon were placed in each of 24 90-L tanks. Treatments
were randomly assigned to the 24 tanks resulting in 4 replicates per dietary
treatment. Four fish from each tank were sampled after 0, 4 and 8 weeks for
weight, length, liver weight, condition factors, hepatosomatic indices, hemocrit,
histopathology, and selenium measurement in liver, kidney, muscle and gill
tissues. 8 fish after 0 and 8 weeks were sampled for whole body selenium
measurement.
Sturgeon survival did not differ significantly among treatment groups after the 8-
week exposure with a mean survival rate of 99 across all groups. Fish fed 41.7 to
191.1 mg Se/kg dw exhibited significant declines in body weight (see table). All
other endpoints measured were as sensitive or less sensitive to selenium in the
diet as body weight.
Mean (SE) white sturgeon moisture, lipid and whole body Se after 8-week exposure
Treatment
group
0.4
9.6
20.5
41.7
89.8
191.1
Moisture, % ww
76.8 (0.5) b
77.0 (0.7) b
76.8 (0.3) b
77.3 (0.5) b
78.5 (0.3) ab
80.0 (0.4) a
Lipid, % ww
9.5 (4) abc
9.5 (0.9) abc
10.1 (0.4) ab
9.6 (0.7) abc
7.6 (0.4) bed
6.1 (0.4) cd
muscle Se, mg/kg dw
8.2 (0.6) e
17.2 (0.7) d
22.9 (1.5) c
36.8 (1.8) b
52.9 (3 .2) a
54.8 (2.8) a
whole body Se, mg/kg dw
5. 2 (0.4) c
11.8 (0.9) b
14.7 (0.8) b
22.5 (1.4) a
34.4 (2.3) a
27.5 (4.4) a
Do not distribute, quote or cite
C-2
Draft Document
-------
Mean (SE) white sturgeon body weight increase after 8-week exposure
Treatment
group
0.4
9.6
20.5
41.7
89.8
191.1
Body weight
increase (%)
282.9 (4.6) a
285.5 (9.9) a
277.7 (6.1) a
191.0 (12.6) b
106.5 (5. 8) c
28.6 (3.6) d
muscle Se, mg/kg dw
8.2 (0.6) e
17.2 (0.7) d
22.9 (1.5) c
36.8 (1.8) b
52.9 (3 .2) a
54.8 (2.8) a
whole body Se, mg/kg dw
5.2 (0.4) c
1 1.8 (0.9) b
14.7 (0.8) b
22.5 (1.4) a
34.4 (2.3) a
27.5 (4.4) a
Letters denote statistical groupings among treatments within each exposure period (p<0.05).
Chronic Value:
Using the logistic equation with a log transformation of the exposure
concentrations (TRAP program), the ECio and EC2o values for reduction in body
weight are 15.08 and 17.82 mg Se/kg dw whole body and 27.76 and 32.53 mg
Se/kg dw muscle tissue.
Do not distribute, quote or cite
C-3
Draft Document
-------
White sturgeon (Tashjian et al 2006)
350
300
s£ 250
CD"
CO
2 200
o
c
S 150
-o
o
-° 100
50
0
.
-
\^
\
\
V
V
3 .9 1.0 1.1 1.2 1,3 1,4 1.5 1.6 1,7 1.8
Log([Se] muscle, mg/kg dw)
Parameter Summary (Logistic Equation Regression Analysis)
Parameter Guess FinalEst StdError 95%LCL 95%UCL
LogXSO 1.6006 1.6303 0.0314 1.5304 1.7301
S 1.6574 2.938 0.925 -0.005 5.882
YO 284.2 286.3 18.9 226.1 346.5
Effect Concentration Summary
% Effect
50.0
20.0
10.0
5.0
Xp Est
42.69
32.53
27.76
23.98
95%LCL
33.92
21.17
15.63
11.75
95%UCL
53.72
49.99
49.30
48.93
16/19/2009 10:32
MED Toxic Response Analysis Model, Version 1.03
Do not distribute, quote or cite
C-4
Draft Document
-------
White sturgeon (Tashjian et al 2006)
350
300
^ 250
8
CD
£ 200
O
IB 150
-o
-° 100
50
0
.(
-
5 .7 .8 .9 1.0 1.1 1.2 1.3 1.4 1.5 1.6
Log([Se] whole body, mg/kg dw)
Parameter Summary (Logistic Equation Regression Analysis)
Parameter Guess FinalEst StdError 95%LCL 95%UCL
LogX50 1.3403 1.3750 0.0643 1.1702 1.5797
S 2.283 2.794 1.908 -3.277 8.865
YO 284.2 294.2 45.0 151.0 437.3
Effect Concentration Summary
% Effect
50.0
20.0
10.0
5.0
Xp Est
23.71
17.820
15.078
12.926
95%LCL
14.80
6.890
4.160
2.587
95%UCL
37.99
46.090
54.655
64.584
(8/19/2009 10:42
MED Toxic Response Analysis Mode), Version 1.03
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C-5
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Linville, R.G. 2006. Effects of Excess Selenium on the Health and Reproduction of White Sturgeon
(Acipenser transmontanus}: Implications for San Franscisco Bay-Delta. Dissertation. University of
California at Davis.
Test Organism:
Exposure Route:
Test Duration:
Study Design:
Effects Data:
EC10 Calculations:
White Sturgeon (Acipenser transmontanus)
Dietary only
Selenium was added to the treatment in the form of selenized yeast. Selenized
yeast (2.2%; Selenomax®, Ambi Inc.) was added to a commercial salmonid diet
and pelleted with fish oil. For the control diet, the selenized yeast mixture
contained 1.3% selenized yeast and 98.7 tortula yeast. Only selenized yeast was
added to the treatment diet. After pelleting, the diet was allowed to air dry on
drying racks.
Females were fed 0.3% body weight/day the experimental diet for 6 months.
16 adult female white sturgeon (approximately 5 years old, mean weight and fork
length: 22.71 kg and 134.59 cm) were exposed in a freshwater flow through
system to either the control diet (8 females in one tank fed 1.4 mg/kg Se) or
treatment (8 females in a separate tank fed 34 mg/kg Se, Se from selenized yeast)
for 6 months. After the 6 month dietary exposure, females were induced to
spawn and fertilized with non-exposed male milt. Eggs were hatched in jars
keeping eggs from each female separate. For each progeny cohort, 3000 larvae
were randomly distributed into 3 reps for stage 40 (intestinal portion is void of
yolk material, but stomach is not differentiated and is filled with yolk) sampling
and 3 reps for stage 45 (yolk sac absorbed, start exogenous feeding)
sampling. Se and biological measurements were made in each replicate.
No Se effects were observed for length or weight of larvae. Effects were
determined for both edema (Table 1) and skeletal (Table 2) deformities. Edema
and deformities increased with stage. EC 10 calculations are based on the
combined effects of edema and skeletal deformities (Table 3) for stage 45 (Table
4), in response to selenium concentrations in eggs.
The EC 10 for total larval deformities (edema + skeletal) in response to Se
concentrations in eggs was calculated using the threshold sigmoid nonlinear
regression model in TRAP (v. 1.22). Se concentrations were log transformed.
The incidence of total deformities was highest at the highest Se concentration, at
27.78%, or 72.22% normal (Table 4). No deformities were observed at three of
the four lower Se concentrations, and 13.33% deformities were observed at one
of the intermediate concentrations (7.61 mg/kg). As a result, the slope of the
concentration-response (C-R) curve, and the resulting EC 10, is determined by the
single highest Se concentration with the 27.78% effect level. The first four Se
concentrations are defining the pre-toxic threshold (the y-intercept term of the
model equation). Because the EC 10 is based on one value, and because that
value shows a relatively low effect level, EC 10 predictions are sensitive to the
initial estimate of the slope of the falling limb of the C-R curve (Table 5).
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Table 1.
Edema Deformities.
Control
Stage 36
Stage 40
Stage 45
Table 2.
Stage 36
Stage 40
Stage 45
Cohort
C3
C4
C5
C4
C5
C4
C5
Skeletal
Control
Cohort
C3
C4
C5
C4
C5
C4
C5
Edema (%)
0.00(1)
0.00(1)
0.00(1)
0.00 (3)
0.00 (3)
0.00 (3)
0.00 (3)
Deformities.
Larval Se
(mg/kg dw)
2.43
1.69
2.67
1.8
2.88
1.96
2.59
Treatment
Cohort
Tl
T2
T3
Tl
T2
T3
Tl
T2
T3
Edema
(%)
0.00(1)
0.00(1)
6.67(1)
0.00 (3)
4.44 ±2.22 (3)
1.67 ±1.67 (2)
0.00 (3)
15.56± 1.11(3)
0.00 (2)
Larval Se
(mg/kg dw)
11.6
18.4
7.75
11.6
20.4
7.22
12
19.4
7.61
Treatment
Skeletal (%)
0.00(1)
0.00(1)
0.00(1)
1.11 ±1.11 (3)
1.11 ±1.11 (3)
0.00 (3)
0.00 (3)
Larval Se
(mg/kg dw)
2.43
1.69
2.67
1.8
2.88
1.96
2.59
Cohort
Tl
T2
T3
Tl
T2
T3
Tl
T2
T3
Skeletal (%)
0.00(1)
0.00(1)
10.00(1)
0.00 (3)
14.44± 1.11(3)
8.33 ±1.67 (2)
0.00 (3)
21.11± 1.11(3)
13.33 ±3.33 (2)
Larval Se
(mg/kg dw )
11.6
18.4
7.75
11.6
20.4
7.22
12
19.4
7.61
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Table 3. Combined Edema and Skeletal Deformities.
Control
Stage
36
Stage
40
Stage
45
Treatment
Egg Se Larval Se
Cohort Affected (%) (mg/kg) (mg/kg) Cohort Affected (%)
C3
C4
C5
C4
C5
C4
C5
0.00(1) 2.46 2.43
0.00(1) 1.61 1.69
0.00(1) 2.68 2.67
l.llil.ll
(3) 1.61 1.8
l.llil.ll
(3) 2.68 2.88
0.00(3) 1.61 1.96
0.00(3) 2.68 2.59
Tl
T2
T3
Tl
T2
T3
Tl
T2
T3
0.00(1)
0.00(1)
16.67(1)
0.00 (3)
18.89± 1.11
(3)
10.00 ±0
(2)
0.00 (3)
27.78 ±2.94
(3)
13.33 ±3.33
(2)
EggSe
(mg/kg)
11
20.5
7.61
11
20.5
7.61
11
20.5
7.61
Larval Se
(mg/kg)
11.6
18.4
7.75
11.6
20.4
7.22
12
19.4
7.61
Table 4. Stage 45 data combined effects - for TRAP input.
Cohort Egg Se (mg/kg) Larval Se (mg/kg) Deformed (%)
Normal (%)
C4
C5
T3
Tl
T2
1.61
2.68
7.61
11
20.5
1.96
2.59
7.61
12
19.4
0
0
13.33
0
27.78
100
100
86.67
100
72.22
Table 5. Effects of initial guess for slope on EC10, EC90/EC10, and goodness of fit.
Initial Slope
2
4
6
9.59
Final Slope
2.63
3.88
5.64
8.75
EC10 (mg/kg)
16.27
17.52
18.40
19.13
EC90/EC10
2.63
1.93
1.57
1.34
Residual Sum of Squares
0.0133
0.0133
0.0133
0.0133
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Chronic Value:
For all EC 10 estimates, a prediction for the initial slope was made, but no initial
guess was made for the logXSO (the concentration representing the EC50 effect
level) of the y-intercept. These values were stable across a range of initial
conditions. The goodness of fit of each model was evaluated using residual sums
of squares. Finally, the EC90/EC10 for each model was calculated.
When TRAP is allowed to make the initial guess for the slope, a slope of 2.645 is
selected. When an initial slope equal to 2.645 or lower is selected, TRAP will
converge on a final slope at or approximate to 2.645 (Table 5). When an initial
guess for a slope greater than 2.645, but equal to or less than 9.59, TRAP will
converge on an increasingly steeper slope, to a maximum of 8.75. However, the
goodness of fit of the resulting model is identical because the uncertainty within
the effects data resulting from having only one point determining the slope of the
falling limb allows TRAP to fit multiple curves across a range of slopes directly
through the highest value (see figures la-Id). The range of EClOs that can be
calculated from TRAP models with the same overall goodness of fit is between
16.27-19.13 mg/kg (Table 5). At slopes of 9.6 or greater, TRAP is no longer able
to find a solution that will pass through the highest value, and the resulting model
fits are poor.
The chronic value for combined deformities is an EC10 of 16.27 mg egg/kg dw,
using the most conservative EC 10 across the range of model results that are
statistically equivalent based on residual sum of squares.
1.0
.4 .6 .8 1.0
Log(mg Se/kg egg dw)
1.2
1.4
Parameter Initial Final
LogXSO 1.416 1.4215
S 2 2.6306
YO 0.96667 0.96667
Figure l.a. Initial estimate for slope set equal to or less than 2.645 (set to 2 for this figure).
EC10 = 16.27 mg/kg.
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1.0
.4 .6 .8 1.0
Log(mg Se/kg egg dw)
1.4
Parameter Initial Final
LogXSO 1.416 1.3863
S 4 3.8754
YO 0.96667 0.96667
Figure l.b. Initial estimate for slope set to 4. EC10 = 17.51 mg/kg.
.4 .6 .8 1.0
Log(mg Se/kg egg dw)
1.4
Parameter Initial Final
LogXSO 1.4159 1.3629
S 6 5.643
YO 0.96667 0.96668
Figure I.e. Initial estimate for slope set to 6. EC10 = 18.40 mg/kg.
Do not distribute, quote or cite
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1.2
1.0
CO
E
o
Ql
.4 .6 .8 1.0
Log(mg Se/kg egg dw)
Parameter
LogXSO
S
YO
Initial
1.416
9.59
Final
1.3447
8.7529
0.96667 0.96667
1.2
1.4
Figure l.d. Initial estimate for slope set to 9.6. EC10 = 19.13 mg/kg.
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Teh, S.J., X. Deng, D-F Deng, F-C Teh, S.S.O. Hung, T.W. Fan, J. Liu, R.M. Higasi. 2004 Chronic
effects of dietary selenium on juvenile Sacramento splittail (Pogonichthys macrolepidotus). Environ. Sci.
Technol. 38: 6085-6593.
Test Organism:
Exposure Route:
Dietary Treatments:
Test Duration:
Study Design:
Effects Data:
Sacramento splittail (Pogonichthys macrolepidotus); juveniles 7-mos.old
Dietary only
8 graded levels of dietary Se; dietary levels obtained by combining selenized
yeast with Torula (non-active) yeast. Selenized yeast contained approximately
21% of Se as selenomethionine and proteinaceous Se forms. Diet was formulated
as pellets by mixing dry ingredients with water and oil, fan-dried, crumbled and
sieved. Analyzed levels: 0.4 (no selenized yeast), 0.7, 1.4, 2.7, 6.6, 12.6, and 57.6
mg/kg.
Fish were fed twice daily with a daily feeding rate of 3% BW in first 5 months
and then adjusted to 2% BW thereafter.
9 months
A flow-through system with 40 fish/tank (24 total tanks) was used; each tank
held 90 L. Flow rate was 4 L/min. Water temperature was maintained at 23°C for
6 months and then 18°C for last 3 months due to failure of water heating system.
5 fish were sampled from each tank at 5 and 9 months and measured for gross
deformities, length, weight, Se in liver and muscle. Sections of the liver were
kept for histopathology. Condition factor (100 x BW/length), heptatosomatic
index (100 x liver weight/BW), BCF (total organ Se/dietary Se) were determined.
Mortality was observed in the two highest dietary treatments: 10 and 34.3%,
respectively. No mortalities were observed in fish fed diets # 12.6 mg/kg. No
significant difference in growth offish fed 12.6 mg/kg Se in diet, but there was in
the fish fed 26.6 mg/kg Se. See table below for levels of Se in fish at 9 months
and associated effects.
Authors determined prevalence of deformities was higher in fish fed 6.6 and 12.6
mg/kg Se in their diet, however a dose-response relationship did not occur (e.g.,
no deformities in high concentration). Gross pathology was a more sensitive
endpoint than growth.
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Summary of effects and assoc. dietary and tissue concentrations in Sacramento splittail
after 9 month exp.
Dietary conc'n mg/kg
Se in liver, mg/kg dw
Se in muscle, mg/kg dw
0.4
20.1
6.6
0.7
18.6
6.9
1.4
20.0
9.2
2.7
23.0
10.1
6.6
26.8
15.1
12.6
31.3
18.9
26.0
40.4
29.4
57.6
73.7
38.7
Liver histopathology (mean lesions scores, N=15)
Macrophage aggregate
Glycogen depletion
Single cell necrosis
Fatty vacuolar degerneration
Eosinophilic protein droplets
Sum of mean lesion scores
0.13
0
0
0
0
0.13
0.07
0
0
0
0
0.07
0.2
0.2
0
0
0
0.4
0.27
0
0.07
0.2
0
0.54
0.40
0.4
0.13
0.53
0
1.46
0.20
0.2
0
0.07
0
0.47
0.20
0
0.07
0.2
0.07
0.54
0.85
1.38
0.46
0.08
0.85
3.62
Gross Pathology (No. of deformities, N=15)
Facial deformities (eye, jaw, and mouth)
Body deformities (kyphosis, lordosis,
scoliosis)
Prevalence of deformity (%)
0
0
0
1
0
6.7
0
4
26.7
1
2
20
5
3
53.3
3
1
26.7
0
1
6.7
0
0
0
Chronic Value:
Comments:
Using gross pathology as the endpoint (prevalence of deformities, %), the
NOAEC is 10.1 mg Se/kg dw and the LOAEC is 15.1 mg/kg Se dw in muscle
tissue; MATC or CV = 12.34 mg/kg Se in muscle dw.
The above concentrations in juvenile muscle tissue cannot be exactly translated
into an equivalent egg-ovary or whole-body concentration in adult splittail. But
using the median egg-ovary to muscle ratio of 1.59 for the family Cyprinidae, the
NOEC and MATC would represent 16.1 and 19.6 mg Se/kg egg-ovary. Using the
median muscle to whole-body ratio of 1.26 for the family Cyprinidae, the NOEC
and MATC would represent 8.04 and 9.83 mg Se/kg whole body. However,
appropriateness of these conversion estimates rests upon uncertain assumptions
that the muscle concentrations in juvenile splittails equal those of adult splittails
under the same exposure conditions, and that splittail tissue ratios are those
typical of the family Cyprinidae.
The authors observed deformities including spinal deformities using fish that
were 7-months-old at test initiation. This is the only study in which deformities
were observed in fish that were not exposed maternally.
Deng et al. (2008) exposed Sacramento splittail juveniles (21-day post hatch) to
dietary selenium and dietary methylmercury in a two factorial design for four
weeks. No adverse effects (growth, condition factor, lethargy or abnormalities)
were observed in the selenium only exposures. The splittail accumulated
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approximately 3.5 mg Se/kg ww muscle in the highest dietary exposure (35 mg
Se/kg. Using the average percent moisture in fish muscle of 78.4% (May et al.
2000), the dw Se concentration is 16.2 mg Se/kg muscle indicating the
recommended CV does not over-estimate an effect concentration.
Rigby et al. (2010) re-analyzed the juvenile Sacramento splittail data generated in
the Teh et al. (2004) study. The authors used logistic regression to estimate EC
values for deformities on a culled data set which eliminated the three highest
dietary treatments due to their departure from a standard concentration-response
relationship. The ECio value for the culled data set was 7.9 mg Se/kg dw muscle
which is lower than the recommended CV of 12.3 mg Se/kg dw muscle. Due to
the lack of a concentration-response relationship across the entire dietary range
and the lack of effects in the Deng et al. (2008) study, an ECio of 7.9 mg Se/kg
dw muscle is too uncertain for a recommended CV. Although the recommended
CV of 12.3 mg Se/kg dw muscle is based on deformities (an uncertain response),
it is considered representative of an effect level for this species because of the
significant reductions in growth at the two highest test concentrations.
Do not distribute, quote or cite C-14 Draft Document
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Bennett, William N., Arthur S. Brooks, and Martin E. Boraas. 1986. Selenium uptake and transfer in
an aquatic food chain and its effects on fathead minnow larvae. Arch. Environ. Contam. Toxicol. 15:513-
517.
Test Organism:
Exposure Route:
Test Duration:
Study Design:
Fathead minnow (Pimephalespromelas; 2 to 8 day-old larvae).
Dietary only
Green alga, Chlorellapyrenoidosa were exposed to Se (H275SeO4) in culture
water for 3 days. Rotifers, Brachionus calyciflorus, were cultured in chambers
with selenium containing green algae at the ratio of 25 |o,g algae/ml to 50 |o,g
rotifer/ml for 5 hr. The rotifers were filtered to separate them from the algae and
immediately heat-killed. The Se concentration in the rotifers was measured for
75Se activity.
9 to 30 days
Selenium uptake by larval fathead minnows was measured in three experiments.
Se-contaminated and control rotifers for feeding to larval fish were prepared in
advance using the low algae:rotifer ratio. Daily equal volumes of rotifers were
divided among five 800 mL polypropylene larval chambers. Three chambers
received Se-contaminated rotifers and two received control rotifers. The rotifers
were dead at the time of feeding (heat killed).
Larval fish were hatched from eggs spawned in the laboratory. After hatching,
active larvae were divided equally among the larval test chambers (daily renewal
exposures using dechlorinated Lake Michigan water). Larvae were initially fed
rotifers raised on control algae (no selenium). The age of the larvae when first
fed Se-contaminated rotifers was 4, 9, and 3 days post-hatch for experiments 1, 2,
and 3, respectively. Larval fish were fed Se-contaminated rotifers for 7, 9, and 7
days in the 3 experiments. A post-exposure observation period of 19 and 2 days
was used for experiments 1 and 2, respectively. During this time the larvae were
fed control rotifers. Daily, larvae from a replicate were removed from the test
chamber, washed, placed in a 20 ml vial, and counted for 75Se activity for 20 min.
All larvae were then placed in test chambers with fresh food rations. At the end
of the study all fish were individually dried and weighed.
Initial feeding of control diet
(days)
Day Se diet first fed
Day Se diet last fed
Observation days on control diet
Age at study termination (days)
Experiment 1
3
4
11
19
30
Experiment 2
8
9
17
2
19
Experiment 3
2
3
9
0
9
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Effects Data:
Mean food Se concentration
(mg/kg)
Food intake (|o,g rotifers/larva)
Initial larvae mean dry wt. at start
of Se-laden food (|o,g)
Final larvae mean dry wt. (|o,g) at
end of test
Final mean larval Se content (|o,g
Se/larva)b
Final mean larval Se
concentrations (mg Se/kg dw)
Experiment 1
>70
50
90
1470 (Control)
800 (Treatment)3
0.0062
43.0
Experiment 2
68
1330
400
1888 (Control)
1354 (Treatment)3
0.0700
51.7
Experiment 3
55
1190
100
475 (Control)
4 16 (Treatment)
0.0248
61.1
a Significantly different from the control.
b Values when Se-laden feeding was ended.
Chronic Value:
Selenium was measured in the test water during the feeding exposures, but the
concentrations were insignificant (0.84 ng/L). Survival was not affected by the
selenium exposures. Preliminary tests showed that fathead minnow larvae would
reach plateau concentrations of selenium within the 7- to 9-day exposure periods.
The food supply was sufficient to sustain growth of the larvae during the study,
according to the authors. The authors state that selenium uptake and higher
selenium content in experiment 2 larvae was due to their larger size and ability to
consume more rotifers/unit time. Se-exposed larvae were significantly smaller
(p<0.05) in mass than controls for experiments 1 and 2.
GM of mean larval Se concentrations measured in the three experiments, i.e.,
43.0, 51.7, and 61.1 mg/kg dw WB, respectively, is 51.40 mg Se/kg dw.
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Dobbs, M.G., D.S. Cherry, and J. Cairns, Jr. 1996. Toxicity and bioaccumulation of selenium to a
three-trophic level food chain. Environ. Toxicol. Chem. 15:340-347.
Test Organism:
Exposure Route:
Rotifer (Brachionus calyciflorus}, and fathead minnow (Pimephales promelas)
12 to 24 hr-old at start.
Dietary and waterborne
Water
Filtered and sterilized natural creek water supplemented with nutrients (Modified
Guillard's Woods Hole Marine Biological Laboratory algal culture medium) for
algal growth. Sodium selenate (Na2SeO4) was added to test water to obtain
nominal concentrations of 100, 200, or 400 (ig Se/L. Concentrations remained
stable and equal in each trophic level.
Control Diet
No selenium was added to the water medium for the alga; green alga was free of
selenium for the rotifer; and rotifers were free of selenium for the fathead
minnow.
Selenium Diet
Sodium selenate was added to the culture medium for the alga; green alga
thereby contained a body burden for the rotifer; and rotifers thereby contained a
body burden for the fathead minnow.
Dietary Treatments: Each trophic level had a different treatment. The green alga was exposed directly
from the water (1, 108.1, 204.9, 397.6 (ig total Se/L); rotifers were exposed from
the water (1, 108.1, 204.9, 393.0 (ig total Se/L) and the green alga as food (2.5,
33, 40, 50 mg Se/kg dry wt); and the fathead minnow were exposed from water
(1, 108.1, 204.9, 393.0 (ig total Se/L) and the rotifer as food (2.5, 47, 53, 60 mg
Se/kg dry wt.).
Test Duration:
Study Design:
25 days
A flow-through system utilizing a stock solution of filtered and sterilized creek
water controlled at 25°C was used to expose three trophic levels of organisms.
Approximately one liter of media was pumped from the algal chamber into the
rotifer chamber each day. A cell density between 3 and 6 x 106 cells/ml was
delivered to the rotifer chambers. Rotifers were started at a density of 151.4 ± 7.7
females/ml and one liter/day of rotifers containing culture water was
intermittently pumped into the minnow chamber. (B. calyciflorus has a life span
of about 7 days at 25°C.) The pump was necessary to overcome the swimming
ability of rotifers to avoid an overflow tube. Larval fathead minnows
(35/chamber) were prevented from escaping by a screened overflow. Chambers
were cleaned daily and aeration was provided. All chambers were duplicated for
test replication and water was measured for selenium on days 0, 2, 6, 7, 11, 14,
17, 20, and 24. All algal and rotifer biomass and selenium samples were made on
these days. Fathead minnow chambers were measured for biomass, dissolved
selenium, and tissue selenium concentrations of days 0, 7, 11, 14, 20, and 24.
Do not distribute, quote or cite C-17 Draft Document
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Effects Data:
Additional measurements were made in the 200 (ig Se/L test chambers on the
fathead minnow on day 16. Selenium concentrations were maintained near the
nominal concentrations and the standard deviation of mean concentrations was
less than 4 percent.
Rotifers. Rotifers did not grow well and demonstrated reduced survival at all
selenium exposure concentrations during the 25 day test. By test day 7 only the
lowest test concentration (108.1 ug/L) had surviving rotifers which showed a
decrease in selenium content from test days 18 through 25. A reduction in rotifer
biomass was discernable by test day 4 in the selenium treatments and since all
test concentrations had viable rotifer populations present, the effect level was
calculated using these data.
Effect of Dietary and Waterborne Selenium on Rotifers after 4 Days Exposure
Se in water, jig/L
1
108.1
202.4
393
Se in diet, nig/kg dw
2.5
33
40
50
Se in rotifer tissue,
mg/kg dw
2.5
40
54
75
rotifer biomass, mg/ml
dw
0.028
0.025
0.011
0.003
Fathead minnows. Due to the reduction of rotifer biomass in the higher test
concentrations, fish mortality and reduction in fish growth observed in the latter
days of the test was difficult to discern between effects from starvation and
selenium toxicity. The data from test day 8 was selected for determining the
effect of selenium on fathead minnows because starvation could be excluded as a
variable.
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Effect of Dietary and Waterborne Selenium on Larval Fathead Minnows after 8 Days Exposure
Se in water, jig/L
1
108.1
202.4
393
Se in diet, nig/kg dw
2.5
47
53
60
Se in fathead minnow
tissue, mg/kg dw
2.5
45
75
73
Average fish weight,
mg dw
0.8
0.7
0.4
0.2
Chronic Value:
Rotifers
Fish
42.36 mg Se/kg dw (EC20)
< 73 mg Se/kg dw (LOAEC) - not amenable to statistical treatment; the LOAEC
was based on the observation that a >50 percent reduction in mean fish weight
occurred at this tissue concentration.
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C-19
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Schultz, R. and R. Hermanutz. 1990. Transfer of toxic concentrations of selenium from parent to
progeny in the fathead minnow (Pimephalespromelas). Bull. Environ. Contam. Toxicol. 45:568-573.
Test Organism:
Exposure Route:
Study Design:
Effects Data :
Chronic Value:
Fathead minnow (Pimephales promelas; Adults)
Dietary and waterborne
Selenite was added to artificial streams which entered the food web; thus, fish
were also exposed to selenium in the diet.
Four Monticello artificial streams were used for the study which lasted from
September 1987 to September 1988. For each study, two streams (treated) were
dosed continuously to achieve 10 |o,g/L and two streams served as controls. Mean
selenium concentrations at the head of the treated streams were 9.8 ±1.2 and
10.3 + 1.7 |og/L, respectively. The concentrations of selenium measured in the
water from controls streams were all less than the detection limit, i.e., 2 (ig/L.
Spawning platforms were submerged into each stream. One subset of six embryo
samples (n = 2000 embryos per sample) were collected from the streams for
selenium analysis. Another subset often embryo samples were reared in
incubation cups receiving the same stream water dosed with sodium selenite via a
proportional diluter. The treated embryos in egg cups received an average 9.7 +
2.6 (ig Se/L. Samples of hatched larvae were analyzed for selenium content while
others were inspected for occurrence of edema and lordosis. Prior to test
termination, female parents were seined. The mean selenium content in the
ovaries of seven to eight females from the treated and control streams was
reported.
Edema and lordosis occurred in approximately 25 percent of the fish spawned
and reared in 10 (ig Se/L. Corresponding occurrence in control fish incubated in
the egg cups was only 1 and 6 percent, respectively. Selenium residues in the
ovaries of females from the control and treated streams were 0.77 and 5.89 mg
Se/kg ww. Using 75.3 percent moisture content in the eggs/ovaries (average
value for fathead minnow ovaries and eggs from GEI Consultants 2008 and
Rickwood et al. 2008), these concentrations equate to 3.12 and 23.85 mg Se/kg
dw.
The LOEC for egg/ovary is <23.85 mg Se/kg dw.
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Beyers, D.W. and Sodergren, C. 200 la. Evaluation of interspecific sensitivity to selenium exposure:
Larval razorback sucker versus flannelmouth sucker. Larval Fish Laboratory. Department of Fishery and
Wildlife Biology, Colorado State University, Fort Collins, Colorado.
Test Organism:
Exposure Route:
Study Design:
Effects Data :
Chronic Value:
Larval flannelmouth sucker (Catostomus latipinnis) and larval razorback sucker
(Xyrauchen texanus)
Dietary and waterborne - laboratory exposure (28-d early life stage)
Continuous flow diluter supplied a range of aqueous test concentrations <1, 25.4,
50.6, 98.9, and 190.6 (ig/L selenate. Well water was used as the dilution water.
Across the range of aqueous exposure concentrations, each test chamber was fed
the same daily ration of living rotifers containing selenium at <0.702, 1.35, 2.02,
4.63, and 8.24 mg/kg dw, respectively. Rotifers accumulated selenium from
algae (Chlorella vulgaris) exposed to 0, 25, 50, 100, and 200 :g/L selenate.
Replicated (n=4) exposure beakers using a randomized, balanced 5x2 factorial
design (1st factor - selenium; 2nd factor - species). Survival was monitored daily
and growth measured at the end of the 28-day exposure. Selenium was measured
in the larvae at the end of the 28-day exposure.
No survival effects were observed and there were no decreases in fish weight or
length. Fish mass was found to increase as a function of selenium concentration.
The chronic values for the flannelmouth sucker and razorback sucker were >10.2
and >12.9 mg Se/kg dw, respectively, based on the concentrations of selenium
measured in whole-body tissue of larval fish at the highest water and dietary
selenium concentrations.
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Beyers, D.W. and Sodergren, C. 200 Ib. Assessment of exposure of larval razorback sucker to selenium
in natural waters and evaluation of laboratory-based predictions. Larval Fish Laboratory. Department of
Fishery and Wildlife Biology, Colorado State University, Fort Collins, Colorado.
Test Organism:
Exposure Route:
Study Design:
Effects Data:
Chronic Value:
Larval razorback sucker (Xyrauchen texanus)
Dietary and waterborne - laboratory exposure (28-d early life stage)
Larvae were exposed in a daily static-renewal system to control water
(reconstituted very hard) and site waters: De Beque, Orchard Mesa, North Pond
diluted 50%, and North Pond. Each water type received either a control diet
(rotifers) or a diet previously exposed to the site water (site food: rotifers fed
algae exposed to respective site water).
Replicated (n=4) exposure beakers using a randomized, balanced 5x2 factorial
design (1st factor - test water type; 2nd factor - rotifers cultured in control water or
in site water). Survival was monitored daily and growth measured at the end of
the 28-day exposure. Selenium was measured in the larvae at the end of the 28-
day exposure.
No survival effects were observed. There were no significant decreases in growth
offish exposed to both site water and site food compared to fish exposed to
control water and control food. There was a significant increase in growth offish
exposed to site water and control food relative to fish exposed to control water
and control food (p<0.0001). There were reductions in the growth offish (14%)
exposed to site water and site food compared to site water and control food
(p<0.0001). Due to the lack of a dose-response relationship in both the
concentration of selenium in the food (rotifers) and growth, and the concentration
of selenium in the fish larvae and growth, the authors did not attribute the effect
of site food on the growth offish to selenium.
The NOAEC for the razorback sucker larvae in the four site water types based on
selenium in whole-body tissue were: De Beque >5.45 mg Se/kg dw; Orchard
Mesa >11 mg Se/kg dw; North Pond 50% dilution >41.1 mg Se/kg dw; North
Pond >42 mg Se/kg dw. Because no significant effects were observed in larvae
exposed to North Pond water at >42 mg Se/kg dw whole-body tissue, this value
was selected as the chronic value for the study.
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C-22
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Muscatello, J.R., P.M. Bennett, K.T. Himbeault, A.M. Belknap and D.M. Janz. 2006. Larval
deformities associated with selenium accumulation in northern pike (Esox Indus) exposed to metal
mining effluent. Environ. Sci. Technol. 40:6506-6512.
Test Organism:
Exposure Route:
Test Duration:
Study Design:
Effects Data:
Northern pike (Esox Indus)
Dietary and waterborne - field exposure
Eggs were collected in the field and incubated in the laboratory. The test was
terminated when the majority of the fry exhibited swim-up and had absorbed the
yolk.
The study area was Key Lake uranium milling operation in north-central
Saskatoon. Spawning northern pike were collected from four sites, one reference
(Davies Creek) and three exposure sites, David Creek near-field (high exposure),
Delta Lake (medium exposure), and David Creek far-field (low exposure). The
exposure sites were located approximately 2, 10 and 15 km downstream of the
effluent discharge. Milt and ova were stripped from ripe fish and eggs were
fertilized in the field. Females were saved for metal analysis and age
determination. Subsamples of ova (prior to fertilization) were collected for metal
analysis.
Although the study sites represent open systems where fish can potentially
migrate among sites, radiotelemetry data from tagged adult pike (Muscatello and
Janz, unpublished data) indicate high site fidelity at the "high" and "medium"
exposure sites (lakes). In contrast, the "low" exposure site likely represents pike
that migrated from further downstream sites that were likely of similar Se
exposures as the reference site.
Eggs were incubated using a two-way ANOVA experimental design using water
collected from reference or exposure sites. So, embryos originating from
reference or exposure site females were incubated in either reference or
appropriate exposure water. In addition, embryos from reference site females
were incubated in water from all four study sites. 50 viable embryos from each
individual female were transferred to each of four replicate incubation chambers.
Cumulative time to 50% eyed, 50% hatch and 50% swim-up were determined.
When the majority of the fry exhibited swim-up and had absorbed the yolk, the
remaining fry were preserved and examined for deformities.
Mean egg diameter and fertilization success did not differ among sites.
Cumulative embryo mortality throughout incubations was not significantly
different among the sites ranging from 45 to 60%. There were no significant
differences in the cumulative time to reach 50% eyed embryos, 50% hatch or
50% swim-up among treatments. Difference in the percent total deformities
between test waters used during embryo incubation exposures were not
significant, so the data were combined for each site (see Table below).
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Selenium concentrations in eggs and muscle from female northern pike collected from reference
and exposed sites and associated total deformities in embryos
Site
Davies Creek
Davies Creek
Davies Creek
Davies Creek
Davies Creek
David Creek (far field)
David Creek (far field)
David Creek (far field)
David Creek (far field)
David Creek (far field)
Delta Lake
Delta Lake
Delta Lake
David Creek (near field)
David Creek (near field)
Site ID
Reference
Reference
Reference
Reference
Reference
Low
Low
Low
Low
Low
Medium
Medium
Medium
High
High
Female
1
2
3
4
5
1
2
3
4
5
1
2
3
1
2
[Se] mg/kg dw
Egg
3.45
2.72
3.39
3.72
2.69
3.39
4.07
4.07
4.07
3.4
43.19
24.53
26.14
48.23
N/A*
Muscle
0.86
1.89
2.56
1.34
1.04
1.95
2.04
1.26
2.48
1.26
17
16.52
16.52
47.82
28.72
Total
deformities %
17
2.5
15.51
7.13
10.41
20.32
13.19
15.33
18.83
11.8
37.8
31.71
26.29
39.5
N/A*
* female had no eggs
Significant increases in total deformities (edema, skeletal deformities,
craniofacial deformities and fin deformities) were observed in fry originating
from pike collected at the medium exposure site. Determination of an effect level
for the percent total deformities relative to the concentration of selenium in eggs
or in female muscle tissue was not amenable to analysis by TRAP. One
requirement of TRAP is to have a response greater than 50%, which was not
satisfied with the available data.
When data are not amenable to determining an effect level using a software
program, such as TRAP, one way to estimate the effect level is to make a direct
measurement of effect at an exposure or tissue concentration. For example, if
only a control and one exposure concentration, 10 |og/L, were tested in an acute
toxicity test and there was 100% survival in the control and 35% in the 10 |og/L,
the effect level would be an EC35 of lOjo, g/L. Such an approach was used to
estimate effect in the Muscatello et al. data. Because no significant differences
were observed in either selenium concentrations in eggs or percent total
deformities between the reference and low exposure site, the data from these 10
sites were combined. Similarly, the egg selenium and total deformity data were
combined for the 4 medium and high exposure sites. These means, geometric for
the selenium concentrations and arithmetic for the percent total deformities, are
given in the following table.
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Mean selenium in northern pike egg and effect values for reference and exposure sites
Sites
Reference sites
(includes low exposure)
exposure sites
[Se] in eggs,
mg/kg dw
(geometric mean)
3.462
34.00
Total deformities,
%
(arithmetic mean)
13.20
33.82
Total deformities, %
(accounting for reference
deformities and transformed
to new scale)3
0
23.76
a The % total deformities in the reference and exposed sites were normalized to the reference effect
(13.2%) and then transformed to a new scale (100%). i.e, Abbott's formula.
The percent affected becomes 24% or an EC24 and the effect level is 34.00
Se/kg dw in eggs
Chronic Value:
EC24 = 34.00 mg Se/kg dw in eggs. Note: an ECi0 cannot be estimated with the
data.
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Hamilton, S.J., K.J. Buhl, N.L. Faerber, R.H. Wiedermeyer and F.A. Bullard. 1990. Toxicity of
organic selenium in the diet of chinook salmon. Environ. Toxicol. Chem. 9:347-358.
Test Organism:
Exposure Route:
Chinook salmon (Oncorhynchus tshawytscha Walbaum; swim-up larvae)
Dietary only
Control Diet
Oregon moist pellet diet where over half of the salmon meal was replaced with
meal from low-selenium mosquitofish (1.0 mg Se/kg dw) collected from a
reference site.
Selenium Diet # 1
Oregon moist pellet diet where over half of the salmon meal was replaced with
meal from high-selenium mosquitofish (35.4 mg Se/kg dw) collected from the
San Luis Drain, CA, termed SLD diet.
Selenium Diet #2
Oregon moist pellet diet where over half of the salmon meal was replaced with
meal from low-selenium mosquitofish same as in the control diet, but fortified
with seleno-DL-methionine (35.5 mg Se/kg dw), termed SeMet diet.
Dietary Treatments: Each selenium diet was formulated to contain about 36 mg Se/kg dw as the high
exposure treatment. The remaining treatments were achieved by thoroughly
mixing appropriate amounts of high-exposure treatment diet with control diet to
yield the following nominal concentrations (3, 5, 10, and 18 mg Se/kg dw).
Test Duration:
Study Design:
90 days
Each dietary treatment was fed twice each day to swim-up larvae (n=100) in each
of two replicate aquaria that received 1 L of replacement water (a reconstituted
experimental water that simulated in quality a 1:37 dilution of water from the San
Luis Drain, CA minus the trace elements) every 15 minutes (flow-through
design). Mortality was recorded daily. Growth was evaluated at 30-day intervals
by measuring the total lengths and wet weights of two subsets of individual fish
(n=10x2) held in separate 11.5 L growth chambers within each replicate
aquarium. Tissue samples were collected for whole-body selenium
determinations (dw basis) at 30-day intervals throughout the study; 10, 5, and 2
fish were sampled from each duplicate treatment after 30, 60, and 90 days of
exposure, respectively. Concentrations of selenium measured in water were
below the limit of detection (1.5-3.1 |o,g/L) in all dietary selenium exposure
concentrations.
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Effects Data:
The magnitude of reduced growth was most evident in the weight of the fish,
although total length was significantly reduced in fish fed high Se-laden diets as
well. The effect of increasing dietary selenium on mean larval weight was similar
in both the SLD and seleno-methionine diets.
Effect of San Luis Drain Diet on Growth and Survival of Chinook Salmon Larvae after 60 Days
Se in diet, mg/kg dw
1
3.2
5.3
9.6
18.2
35.4
Se in chinook salmon,
mg/kg dw
0.9
3.3
4.5
8.4
13.3
29.4
Mean larval weight, g
3.35
2.68
2.76
2.8
2.62
1.4
Survival, %
99
97.3
93
95
92.4
89
Effect of Seleno-methionine Diet on Growth and Survival of Chinook Salmon Larvae after 60
Days
Se in diet, mg/kg dw
1
3.2
5.3
9.6
18.2
35.4
Se in chinook salmon,
mg/kg dw
0.9
2
3.1
5.3
10.4
23.4
Mean larval weight, g
3.35
3.08
3.22
3.07
2.61
1.25
Survival, %
99
100
95
94.1
92.4
62.5
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Chronic Value:
Due to unacceptable control mortality of swim-up larvae in control treatments
after 90 days (33.3 percent - SLD diet; 27.5 percent - SeMet diet), chronic values
had to be determined from respective values reported after 60 days (tables
above).
Analysis of the elemental composition of the SLD diet indicated that B, Cr, Fe,
Mg, Ni and Sr were slightly elevated compared to the control and SeMet diets.
No additional analyses were performed to determine the presence of other
possible contaminants, i.e., pesticides.
Diet
type
SLD
SeMet
EC2o values
Survival
(after 60 d of
exposure)
Tissue Se
(mg/kg dw)
NAa
NAa
Growth
(after 60 d of exposure)
Whole body Tissue Se
(mg/kg dw)
15.73
10.47
ECio values
Growth
(after 60 d of exposure)
Whole body Tissue Se
(mg/kg dw)
11.14
7.355
a The EC20 and ECi0 values for survival of swim-up larvae versus levels of selenium for the SLD and
SeMet dietary exposure could not be estimated using non-linear regression.
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Hamilton et al (1990) Chinook Salmon fed SLD Diet
Logistic Equation, Three Parameter Model, Se concentrations logic transformed
LogX50
StDev
YO
% Effect
50
20
10
5
4.U
3.5
4
_ 3.0
o)
I, 2.5
S
- 2.0
>
" 1.5
c
CO
CD
s 1.0
.5
0
(
•
*~* ^*^V
\^
\
^v
\
-
D .2 .4 .6 .8 1.0 1.2 1.4 1.6
Log(Se in Chinook Salmon mg/kg dw)
Guess FinalEst SE 95%LCL 95%UCL
1.453 1.453 7.30E-02 1.2206 1.6854
1.353 1.353 6.67E-01 -7.71E-01 3.4769
2.968 2.968 1.89E-01 2.3651 3.5709
Xp Est 95% LCL 95% UCL
28.379 16.62 48.458
15.734 5.7003 43.431
11.143 2.4771 50.127
8.1085 1.1213 58.637
DF SS MS F P
Total
Model
Error
5 2.0749 0.41498
2 1.8202 0.91009 10.719 0.95699
3 0.2547 8.49E-02
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Chinook salmon SeMet diet (Hamilton et al. 1990)
.2 .4 .6 .8 1.0 1.2
Log([Se]whole body, mg/kg dw)
1.4
Parameter Summary (Logistic Equation Regression Analysis)
Parameter Guess FinalEst StdError 95%LCL 95%UCL
LogXSO 1.3148 1.2823 0.0242 1.2053 1.3593
S 0.6971 1.3214 0.1826 0.7404 1.9025
YO 3.217 3.239 0.067 3.027 3.452
Effect Concentration Summary
% Effect
50.0
20.0
10.0
5.0
Xp Est
19.156
10.472
7.355
5.312
95%LCL
16.045
7.516
4.595
2.899
95%UCL
22.870
14.591
11.775
9.733
06/19/2009 1352
MED Toxic Response Analysis Model, Version 1 03
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C-30
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Hilton, J.W. and P.V. Hodson. 1983. Effect of increased dietary carbohydrate on selenium metabolism
and toxicity in rainbow trout (Salmo gairdneri). J. Nutr. 113:1241-1248.
Test Organism:
Exposure Route:
Test Treatments:
Test Duration:
Study Design:
Effects Data:
Rainbow trout (Oncorhynchus mykiss; juvenile; approx. 0.6 g each)
Dietary only
Low carbohydrate diet (LCD)
This diet contained capelin oil at 11 percent of the diet with cellulose as the filler.
High carbohydrate diet (HCD)
This diet contained cere lose at 25 percent of the diet with cellulose as the filler.
For both diets, the selenium was supplemented as sodium selenite which was
mixed with cellulose and then added to the diet as a selenium premix.
The two diets were supplemented with selenium (as sodium selenite) at the rate
of 0, 5, or 10 mg/kg dw to make up the six different dietary selenium treatments
(n = 3 low carbohydrate diet; n= 3 high carbohydrate diet). The six diets were fed
to duplicate groups of 100 fish. The trout were fed to satiation 3-6 times per day.
Measured concentrations of selenium in the low carbohydrate diet were: 0.6
(control), 6.6, and 11.4 mg/kg dw, and the measured concentrations of selenium
in the high carbohydrate diet were: 0.7 (control), 6.6, and 11.8 mg/kg dw. The
tanks received a continuous flow of water with a flow rate of 3-4 liters per
minute.
16 weeks
Body weights, feed: gain ratios, and total mortalities were determined after each
28-day interval. After 16 weeks, approximately 20 fish were randomly removed
from each tank, weighed, and blood was collected for hemoglobin, hematocrit,
and plasma glucose, protein, and calcium determination. The livers and kidneys
were then dissected. The livers were assayed for glycogen content, and samples
of both liver and kidney were assayed for selenium content. Additional
subsamples of fish were sacrificed and assayed for selenium content and for ash,
crude protein, and moisture content (n=6 per treatment). Finally, 30 fish were
killed, their livers and kidneys dissected, and analyzed for Ca, Cu, Fe, Mg, P, and
Zn content.
The only overt sign of selenium toxicity was food avoidance observed in trout
fed the highest selenium content in both low and high carbohydrate diets, which
led to significantly reduced body weight after 16 weeks. There were no
significant differences detected between treatment groups in hematological
parameters. Kidney, liver, and carcass selenium levels increased with increasing
selenium content of the diet, however, only the liver selenium concentrations
were significantly affected by dietary selenium level, dietary carbohydrate level,
and the interaction between the two treatments. Mineral analysis of the kidney
showed significantly higher levels of calcium and phosphorous in trout reared on
the two highest levels of dietary selenium. Concentrations of copper in the liver
increased significantly with increasing dietary selenium levels and decreasing
dietary carbohydrate levels.
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Effect of Selenium in Low carbohydrate Diet to Rainbow Trout
Se in diet, nig/kg dw
0.6
6.6
11.4
Se in trout liver, mg/kg dw
0.8
38.3
49.3
Trout weight, kg/100 fish
3.3
3.3
1.8
Effect of Selenium in High carbohydrate Diet to Rainbow Trout
Se in diet, mg/kg dw
0.7
6.6
11.8
Se in trout liver, mg/kg dw
0.6
21.0
71.7
Trout weight, kg/100 fish
2.7
2.3
1.4
Chronic Value:
The following table lists the NOAEC, LOAEC and MATC for both diets in liver
tissue. EC values could not be determined for this study. Data did not meet
minimum requirements for analysis.
Diet
Low carb
high carb
NOAEC, mg Se/kg dw
liver
38.3
21.0
LOAEC, mg Se/kg dw
liver
49.3
71.7
MATC, mg Se/kg dw
liver
43.5
38.8
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Hicks, B.D., J.W. Hilton, and H.W. Ferguson. 1984. Influence of dietary selenium on the occurrence of
nephrocalcinosis in the rainbow trout, Salmo gairdneri Richardson. J. Fish Diseases. 7:379-389.
(Note: These data are the exact same as reported for the low carbohydrate diet in Hilton and Hodson
1983, with the addition of prevalence of nephrocalcinosis occurring in trout after 16 to 20 weeks of
consuming the contaminated test diets).
Test Organism:
Exposure Route:
Test Treatments:
Test Duration:
Study Design:
Effects Data:
Chronic Value:
Rainbow trout (Oncorhynchus mykiss; juvenile; approx. 0.6 g each)
Dietary only
This diet contained capelin oil at 11 percent of the diet with cellulose as the filler.
The selenium was supplemented as sodium selenite which was mixed with
cellulose and then added to the diet as a selenium premix.
The test diet was supplemented with selenium (as sodium selenite) at the rate of
0, 5, or 10 mg/kg dw to make up the three different dietary selenium treatments.
The three diets were fed to duplicate groups of 100 fish. The trout were fed to
satiation 3-6 times per day. Measured concentrations of selenium in the low
carbohydrate diet were: 0.6 (control), 6.6, and 11.4 mg/kg dw. The tanks received
a continuous flow of water with a flow rate of 3-4 liters per minute.
16 to 20 weeks
See Hilton and Hodson (1983). After 20 weeks on the test diets, ten fish were
randomly removed from each treatment. Tissues for histopathological
examination included the stomach, intestine and pyloric ceca (including
pancreas), spleen, liver, heart, kidney, skin, muscle, and gills.
Only effects of selenium on kidney tissue are included in the article. The kidneys
of the 10 trout fed the highest selenium content in the diet exhibited normal
appearance. Five of these trout exhibited precipitation of calcium in the tubules
with some epithelial necrosis, but no loss of epithelial continuity. Extensive
mineralized deposition of Ca within the tubules, tubular dilation and necrosis of
tubular epithelium, ulceration of tubules, and intestinal Ca mineralization was
observed in four of the ten fish.
Same as for growth of rainbow trout reported by Hilton and Hodson (1983). The
MATC estimated for growth of rainbow trout relative to final concentration of
selenium in liver tissue of trout reared on the low carbohydrate diet is the GM of
38.3 (NOAEC) and 49.3 (LOAEC) mg/kg dw, or 43.45 mg/kg dw.
EC values could not be determined for this study. Data did not meet minimum
requirements for analysis.
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Hilton, J.W., P.V. Hodson, and S.J. Slinger. 1980. The requirements and toxicity of selenium in
rainbow trout (Salmo gairdneri). J. Nutr. 110:2527-2535.
Test Organism:
Exposure Route:
Test Duration:
Study Design:
Effects Data:
Rainbow trout (Oncorhynchus mykiss; juvenile; approx. 1.28 g each)
Dietary only
A casien-torula yeast diet was formulated to contain geometrically increasing
levels of selenium from 0 to 15 mg/kg dw. The selenium was supplemented as
sodium selenite which was mixed with cellulose and then added to the diet as a
selenium premix.
20 weeks
Six test diets were fed to triplicate groups of 75 fish. The trout were fed to
satiation 3-4 times per day, 6 days per week, with one feeding on the seventh
day. Measured concentrations of selenium in the diet were: 0.07 (control), 0.15,
0.38, 1.25, 3.67, and 13.06 mg/kg dw. The tanks received a continuous flow of
dechlorinated tap water from the City of Burlington, Ontario municipal water
supply. The waterborne selenium content of this water was 0.4)0, g/L. During the
experiment, the fish were weighed every 2 weeks with the feeding level adjusted
accordingly. Mortalities were noted daily and the feed consumption for each
treatment was recorded weekly. After 4 and 16 weeks, three to six fish were
randomly removed from each tank, sacrificed, and their livers and kidneys
removed and weighed. An additional three to six fish were then obtained from
each treatment, killed, and prepared for tissue analysis. Organs and carcasses
were freeze-dried for determination of selenium concentration. After 16 weeks,
three more fish were removed. Kidney, liver, spleen and dorsal muscle tissue was
dissected for examination of histopathology. At the end of 8 and 16 weeks, four
to five fish were removed, sacrificed, and a blood sample was taken for
hematological measurements (hematocrit, red blood cell count, and blood iron
concentration). After 20 weeks, three to four more fish were removed, sacrificed,
and a blood sample was taken for measurement of glutathione peroxidase
activity.
There were no significant differences detected between treatment groups in
histopathology, hematology, or plasma glutathione peroxidase activity. Trout
raised on the highest dietary level of selenium (13.06 mg/kg dw) had a
significantly lower body weight and a higher number of mortalities (10.7;
expressed as number per 10,000 fish days) than trout from the other treatments
levels after 20 weeks of exposure.
Do not distribute, quote or cite
C-34
Draft Document
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Effects on Juvenile Rainbow Trout
Se in diet, mg/kg dw
0.07
0.15
0.38
1.25
3.67
13.06
Se in Liver, mg/kg dw
0.6
0.95
2.4
11
40a
100b
Weight, g/fish
3.2
3.5
3.7
4.1
4.1
1.4
Mortality*
0
0
0.6
0.6
0
10.7
* expressed as number per 10,000 fish-days
a NOAEC
b LOAEC
Chronic Value:
NOAEC = 40 mg Se/kg dw
LOAEC = 100 mg Se/kg dw
MATC = 63.25 mg Se/kg dw
Do not distribute, quote or cite
C-35
Draft Document
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Holm, J. 2002. Sublethal effects of selenium on rainbow trout (Oncorhynchus mykiss) and brook trout
(Salvelinusfontinalis). Masters Thesis. Department of Zoology, University of Manitoba, Winnipeg, MB.
Holm, J., V.P. Palace, K. Wautier, R.E. Evans, C.L. Baron, C. Podemski, P. Siwik and G. Sterling.
2003. An assessment of the development and survival of rainbow trout (Oncorhynchus mykiss) and brook
trout (Salvelinusfontinalis) exposed to elevated selenium in an area of active coal mining. Proceedings of
the 26th Annual Larval Fish Conference 2003, Bergen, Norway. ISBN 82-7461-059-B.
Holm, J., V.P. Palace, P. Siwik, G. Sterling, R. Evans, C. Baron, J. Werner, and K. Wautier. 2005.
Developmental effects of bioaccumulated selenium in eggs and larvae of two salmonid species. Environ.
Toxicol. Chem. 24: 2373-2381.
Test Organism:
Exposure Route:
Study Design:
Effects Data :
Rainbow trout (Oncorhynchus my kiss; spawning adults) and brook trout
(Salvelinusfontinalis; spawning adults)
Dietary and waterborne - field exposure
Total selenium concentrations measured at the high selenium site ranged from 6
to 32 |og/L. Selenium was not measured at the reference streams; selenium
concentrations at reference locations in the area ranged from <0.5 to 2.2 |og/L.
Spawning fish were collected at low selenium or reference streams (Deerlick
Creek, Wampus Creek and Cold Creek), a slightly elevated selenium stream
(Gregg Creek), and an elevated selenium stream (Luscar Creek) in the
Northeastern slopes region of Alberta, Canada. An active coal mine is the source
of selenium in the elevated streams. Eggs and milt from the spawning trout were
expressed by light pressure from abdomen. Individual clutches of eggs were
fertilized from a composite volume of milt derived from 3-5 males. Fertilized
eggs from individual females were reared to swim-up stage and examined for a
number of parameters including percent fertilization, mortality, edema, and
deformities (craniofacial, finfold, and spinal malformations). Similar studies
were conducted in 2000, 2001 and 2002. One notable difference is that the
embryos were incubated at 8°C in 2000 and at 5°C in 2001. The authors noted
that 5°C is a better representation of the actual stream temperature during embryo
development.
Other than selenium, there were no significant differences in the concentrations
of other elements (Al, As, Sb, Ba, Be, Ni, B, Cd, Ca, Cr, Co, Cu, Fe, Pb, Li, Mg,
Mn, Hg, Mo, Ag, Sr, Tl, Th, Sn, Ti, U, V, Zn) in trout eggs between the low level
and elevated selenium streams. There are two ways to approach determination of
effects due to selenium in this study and both are presented here. The first
approach determines effects based on a comparison of average conditions
between streams (between streams approach). For example, if there is a
significant difference between the average frequency of deformities in a
contaminated stream and reference stream, the effect level for the between
streams approach would be the average concentration of selenium in the tissue
from the contaminated stream. The second approach evaluates individual
response variables (e.g., edema, deformities) against the individual selenium
tissue concentrations for the combined contaminated and reference stream data
Do not distribute, quote or cite
C-36
Draft Document
-------
set with each year (within streams approach). This approach, which results in EC
estimates (e.g., ECio and EC2o) if the data meet the model assumptions, is
explained in the Calculations of Chronic Values section of the text.
Between streams approach: For each sampling location (stream), data for the
three years (Tables 1 and 2) were combined in the between streams analysis of
variance (ANOVA). For rainbow trout embryos, there were no significant
differences in fertilization, time to hatch and mortality between the streams with
elevated selenium and the reference streams. ANOVA indicated significant
differences in the frequency of embryonic effects between streams (Table 3). The
analysis did not prove useful; however, due to a higher occurrence of effects in
some of the reference streams relative to the exposed streams (Tables 3 and 4).
The between streams analysis, therefore, was not used to determine effect
concentrations for rainbow trout.
ANOVA of brook trout data indicated the only significant difference in
embryonic abnormalities among sites was craniofacial deformities (Tables 5 and
6). Significant differences were also found for fertilization and larval weight. The
highest average percent fertilization was observed at the site with the greatest
concentration of selenium in eggs, which indicates that the differences in
fertilization among sites were not caused by variation in selenium concentrations.
Because the percent of embryos with craniofacial deformities in Luscar Creek
was 7.9% (2.1% in Cold Creek), it was not considered biologically meaningful.
Likewise the significantly lower larval weights at the exposed sites was not large
(16% lower than Cold Creek larvae) and again coupled with the low occurrence
of abnormalities by the brook trout, a signature of selenium effects, the lower
larval weights were not considered biologically meaningful.
Within streams approach: As with the between streams analysis, data were
combined for the three years of study in the within streams analysis (Tables 1 and
2). Craniofacial deformities, skeletal deformities and edema in rainbow trout
embryo, as a function of selenium in egg ww, were fitted to a logistic curve from
which ECio and EC20 values were calculated (see table below and Figures 1 and
2). EC estimates for fmfold deformities, length and weight of rainbow trout
embryos could not be made because of inadequate dose-response. The brook
trout data were not suitable for fitting logistic curves (Figure 3).
Do not distribute, quote or cite C-37 Draft Document
-------
Rainbow Trout EC Estimates using TRAP Logistic Equation, log([Se]egg)
Response
100% -
%craniofacial
100% -
%skeletal
100% -
%edema
EC20
Se, mg/kg ww
11.4
11.0
9.9
Se, mg/kg
dwa
29.4
28.4
25.5
EC10
Se, mg/kg ww
10.3
8.2
9.5
Se, mg/kg
dwa
26.5
21.1
24.5
Comment
Large SE for
steepness
ww to dw was converted using 61.2% moisture for rainbow trout eggs (Seilor and Skorupa, 2001)
Do not distribute, quote or cite
C-38
Draft Document
-------
Table 1. Rainbow trout embryo-larval parameters collected from a high Se site (Luscar Creek), an
intermediate Se site (Gregg River), and reference sites (Deerlick Creek and Wampus Creek) in
northeastern Alberta over three consecutive years.
Year
2000
2000
2000
2000
2000
2000
2000
2001
2001
2001
2001
2001
2001
2001
2001
2001
2001
2001
2001
2001
2001
2001
2001
2001
2001
2001
2001
2001
2001
2001
2001
2001
2001
2001
2002
2002
2002
Site Female #
Luscar
Luscar
Luscar
Deerlick
Deerlick
Deerlick
Deerlick
Luscar
Luscar
Luscar
Luscar
Luscar
Luscar
Luscar
Luscar
Luscar
Luscar
Deerlick
Deerlick
Deerlick
Deerlick
Deerlick
Deerlick
Deerlick
Deerlick
Deerlick
Deerlick
Gregg
Gregg
Gregg
Gregg
Gregg
Wampus
Wampus
Luscar
Luscar
Luscar
11
12
14
16
17
18
15
1
3
4
8
14
32
33
39
40
41
8
9
10
16
17
21
22
23
25
39
2
3
5
9
18
9
13
3
8
10
Se in eggs,
mg/kg ww
6.84
6.66
11.6
1.78
1.39
1.00
5.01
5.39
8.39
6.48
4.47
10.4
5.64
3.88
5.14
3.36
11.7
3.68
3.08
1.62
2.62
2.79
1.96
3.13
3.03
3.32
2.43
4.57
4.49
4.05
5.09
5.97
2.66
2.04
5.4
18.3
22
%craniofacial %skeletal %finfold
deformities deformities deformities
7.18
1.48
14.43
0.63
0
0
0
7.35
6.29
22.22
12
34.55
8.24
5.26
1.91
11.62
37.67
9.55
5.39
7.89
24.24
14.13
13.27
1.09
9.65
9.25
11.89
11.97
5.58
4.95
20
16.13
16.07
7.84
60.47
94.12
100
13.26
4.43
23.71
1.9
0
0.86
0
6.76
4.97
22.22
9.33
44.85
5.97
6.58
3.18
7.05
83.41
5.45
4.98
7.89
48.48
15.22
35.71
2.17
14.04
13.29
9.09
7.75
9.3
5.45
13.85
19.35
0
9.8
27.9
23.5
64.3
1.66
0.74
7.22
0.63
0
0
0
3.53
2.98
33.33
2.67
4.24
3.13
9.21
0
5.39
3.59
1.36
0.41
5.26
3.03
4.35
7.14
0
3.51
7.51
7.69
15.49
2.33
2.48
15.38
41.94
1.79
1.31
93
4.4
3.6
%edema
4.97
1.85
85.57
0.63
0
0
0
2.94
6.95
26.67
10.67
43.64
9.09
3.95
1.27
6.64
87
5.45
2.07
10.53
12.12
20.65
25.51
1.09
7.89
8.09
14.69
7.04
4.65
5.94
16.15
35.48
7.14
7.84
14
97.1
100
Do not distribute, quote or cite
C-39
Draft Document
-------
Year Site Female # Se in eggs, %craniofacial %skeletal %finfold
mg/kg ww deformities deformities deformities
%edema
2002
2002
2002
2002
2002
2002
2002
2002
2002
2002
2002
2002
2002
2002
2002
2002
2002
Luscar
Luscar
Luscar
Luscar
Luscar
Deerlick
Deerlick
Deerlick
Deerlick
Deerlick
Deerlick
Gregg
OO
Wampus
Wampus
Wampus
Wampus
Luscar
12
22
23
24
26
10
18
21
24
25
26
1
1
2
3
4
28
15.7
20.5
6.3
26.8
6.5
5.9
7.8
5
4.3
4.4
6.6
5.8
3
4
4.6
4.7
7
82.35
100
5.59
100
1.72
5.65
10.77
6.9
2.88
5.3
2.95
4.76
18.84
0
4.1
25
19.23
47.1
42.1
6.6
100
1.7
7.26
1.54
6.9
2.88
5.3
1.85
3.81
14.49
0
3.28
20
0
66.7
2.1
1.6
0
4.3
7.26
9.23
20.69
21.58
6.82
1.11
3.81
72.46
100
7.58
70
76.9
52.9
100
2.7
100
0.9
3.23
3.08
1.72
0.72
3.03
1.85
3.81
11.59
100
0.61
12.5
0
Do not distribute, quote or cite
C-40
Draft Document
-------
Table 2. Brook trout embryo-larval parameters collected from a high Se site (Luscar Creek), an
intermediate Se site (Gregg River), and reference site (Cold Creek) in northeastern Alberta over
three consecutive years.
Year Location Female # Se in egg, mg/kg %craniofaci %skeletal %finfold %edema
2000
2000
2000
2000
2000
2000
2000
2000
2000
2000
2000
2000
2000
2000
2000
2000
2000
2000
2001
2001
2001
2001
2001
2001
2001
2001
2001
2001
2001
2001
2001
2001
2001
2001
2001
2001
2001
2001
Luscar
Luscar
Luscar
Luscar
Luscar
Luscar
Luscar
Luscar
Luscar
Luscar
Luscar
Cold
Cold
Cold
Cold
Cold
Cold
Cold
Cold
Cold
Cold
Cold
Cold
Luscar
Luscar
Luscar
Luscar
Luscar
Luscar
Luscar
Luscar
Luscar
Luscar
Luscar
Gregg
Gregg
Gregg
Gregg
Do not distribute, quote
1
2
3
5
12
13
14
15
16
17
18
21
22
24
25
26
33
34
6
7
8
21
51
3
7
17
19
59
60
61
64
76
82
83
3
22
23
25
or cite
4.78
4.83
5.98
3.86
6.06
5.8
5.17
9.92
5.03
6.01
12.7
1.15
1.83
0.97
No data
0.59
1.35
2.18
1.79
1.36
0.94
1.07
1.09
8.4
7.26
14.6
9.79
5.8
9.03
7.29
7.08
7.1
6.06
5.82
7.08
7.95
9.23
6.46
C-41
15.38
38.06
7.39
25
16.77
4.06
4.13
16.22
5.61
9.44
14.34
3.26
4.83
1.67
3.31
3.45
6.15
6.45
0
1.61
1.36
0.43
0
0
1.35
2.22
7.55
2.28
3.16
0
1.54
36.71
1.11
6
6.32
0
0.5
0.56
0
1.49
3.03
5.7
1.83
1.42
0.49
0.54
0
5.83
0.72
1.48
1.38
0
1.1
4.83
0
0
0
0.69
0
0
2.13
0.93
1.62
0.63
2.11
0.46
0
0
2.19
13.29
0.22
2
1.58
0
0.5
0
0
3.73
0.34
8.77
0.7
0.2
0.36
0.54
0.27
0.83
0
0.89
1.38
0.72
1.66
6.9
1.54
0.81
0
0.46
0.27
0
0
0
0.81
0.32
2.42
0.91
1.05
9.09
0
19.65
0.88
5.6
20.53
1.08
2.51
0.56
D
15.38
1.49
0.5
4.82
0
0
0.12
0
0.27
1.11
0.36
0
0.69
0
1.1
0.69
0
0
0
1.38
0.54
0
6.38
0.46
0.27
0
0.3
0.46
1.05
0
0
1.16
0.44
0.8
1.58
0
0
0
raft Documer
-------
Year Location Female # Se in egg, mg/kg %craniofaci %skeletal %finfold %edema
2001
2001
2001
2001
2002
2002
2002
2002
2002
2002
2002
2002
2002
2002
2002
2002
2002
2002
2002
2002
2002
2002
2002
2002
2002
Gregg
Gregg
Gregg
Gregg
Luscar
Luscar
Luscar
Luscar
Luscar
Luscar
Luscar
Luscar
Gregg
Gregg
Gregg
Cold
Cold
Cold
Cold
Cold
Cold
Cold
Cold
Cold
Cold
31
32
33
34
17
23
26
38
42
44
54
56
25
37
39
32
26
2
5
29
23
48
42
22
51
7.35
4.91
7.02
5.01
6.28
5.27
6.36
18.9
4.95
6.47
7.96
18.8
6.27
4.58
6.67
0.42
0.89
0.94
1
1.02
1.2
1.25
1.6
1.74
2.11
0.51
7.21
1.88
0
1.7
7.34
1.81
0.9
2.79
0
0.33
3.99
1.23
2.99
3.57
0
0
0.96
0.25
0.72
0.35
9.52
0
0
2.17
1.7
0.48
1.88
0.37
12.74
0.46
0.52
0.54
0.44
0.25
0.33
0.75
1.23
0
1.19
0.6
0
0.32
0.5
1.09
0.35
4.76
0
0
2.17
0.17
3.37
4.38
0
0.85
0
0.26
0
0.15
0
0
0.5
0
0
1.19
0
0
0
0.25
0.36
0.35
2.38
0
1.09
0
0
0.48
0
0
0.21
0.46
0.26
0.18
0.15
0
0
0.75
0
0
1.19
0
0.29
0
0
0.72
0.35
0
0
1.09
2.17
Do not distribute, quote or cite
C-42
Draft Document
-------
Table 3. Results of ANOVA comparing rainbow trout endpoints among sites
% fertilization
Site
Residuals
Df
3
51
Sum of Sq
77.60
20817.33
Mean Sq
25.8653
408.1829
F Value
0.06336703
Pr(F)
0.978935
% mortality
Site
Residuals
Df
3
51
Sum of Sq
3751.51
34510.50
Mean Sq
1250.504
676.676
F Value
1.848008
Pr(F)
0.1502207
% craniofacial deformities
Site
Residuals
Df
3
50
Sum of Sq
8093.97
30449.48
Mean Sq
2697.989
608.990
F Value
4.430272
Pr(F)
0.007732133
% skeletal deformities
Site
Residuals
Df
3
50
Sum of Sq
3279.30
19703.16
Mean Sq
1093.101
394.063
F Value
2.773923
Pr(F)
0.05094422
% finfold deformities
Site
Residuals
Df
3
50
Sum of Sq
6273.17
26886.93
Mean Sq
2091.056
537.739
F Value
3.888612
Pr(F)
0.01417887
% edema
Site
Residuals
Df
3
50
Sum of Sq
8902.51
43012.30
Mean Sq
2967.502
860.246
F Value
3.449597
Pr(F)
0.0233558
Do not distribute, quote or cite
C-43
Draft Document
-------
Table 3. Results of ANOVA comparing rainbow trout endpoints among sites (continued)
Fry length
Site
Residuals
Df
3
50
Sum of Sq
5.0847
148.8246
Mean Sq
1.694896
2.976493
F Value
0.5694271
Pr(F)
0.6377436
Fry weight
Site
Residuals
Df
3
48
Sum of Sq
1721.104
7726.859
Mean Sq
573.7012
160.9762
F Value
3.563888
Pr(F)
0.02080915
Table 4. Rainbow trout means (standard deviation) for measurements made in eggs, embryos and
larvae spawned from fish collected at exposed sites (Luscar and Gregg Creeks) and reference sites
(Deerlick and Wampus Creeks).
Parameter
egg Se, mg/kg ww
fertilization, %
mortality, %
craniofacial, %
skeletal, %
finfold, %
edema, %
larval length, mm
larval weight, mg
Site
Luscar Cr.
9.93 (6.77)
77.8 (20.3)
35.0(29.5)
33.3 (37.2)
25.0(27.9)
15.0(27.1)
34.5 (40.3)
18.5 (2.0)
53.3(16.3)
Gregg Cr.
6.52(4.11)
81.2(12.7)
34.2 (32.5)
10.6 (6.5)
9.9 (5.8)
13.6(15.2)
12.2(12.3)
19.4(1.6)
44.6 (10.4)
Deerlick Cr.
3.49(1.90)
77.5 (20.9)
18.1 (14.6)
7.1(6.1)
9.2(12.3)
5.4 (6.2)
6.1 (7.3)
19.0(1.5)
41.2(9.3)
Wampus Cr.
3.5 (1.09)
77.5(24.1)
37.3 (34.5)
12.0 (9.6)
7.9 (8.2)
42.2 (43.7)
23.3 (37.8)
19.2 (0.9)
40.6 (8.4)
Do not distribute, quote or cite
C-44
Draft Document
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Table 5. Brook trout means (standard deviation) for measurements made in eggs, embryos and
larva spawned from fish collected at exposed sites (Luscar and Gregg Creeks) and
reference site (Cold Creek).
Parameter
egg Se, mg/kg ww
fertilization, %
mortality, %
craniofacial, %
skeletal, %
finfold, %
edema, %
larval length, mm
larval weight, mg
Site
Luscar Cr.
7.78 (3.80)
92.8 (7.2)
6.5 (8.9)
7.9(10.1)
2.0 (3.3)
1.9(4.1)
1.0(2.9)
17.4(1.1)
31.7(8.6)
Gregg Cr.
6.59(1.39)
78.4(18.2)
2.9 (2.3)
2.3 (2.5)
0.8 (0.7)
3.1(6.0)
0.3 (0.6)
17.9(0.9)
31.3(5.4)
Cold Cr.
1.26(0.47)
89.1(19.6)
6.9(12.1)
2.1(2.6)
1.0(1.4)
0.9(1.5)
0.7(1.4)
18.5(1.2)
37.8 (7.2)
Table 6. Results of ANOVA comparing brook trout endpoints among sites
% fertilization
site
Residuals
% mortality
site
Residuals
df
2
60
df
2
60
Sum of Sq
1683.3
12906.4
Sum of Sq
131.4
5433.6
% craniofacial deformities
site
Residuals
df
2
60
Sum of Sq
519.1
3150.6
Mean Sq
841.67
215.11
Mean Sq
65.72
90.56
Mean Sq
259.54
52.51
F Value
3.9128
F Value
0.7257
F Value
4.9427
Pr(F)
0.0253
Pr(F)
0.4882
Pr(F)
0.0103
Do not distribute, quote or cite
C-45
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Table 6. Results of ANOVA comparing brook trout endpoints among sites (continued)
% skeletal deformities
site
Residuals
df
2
60
% finfold deformities
site
Residuals
% edema
site
Residuals
Fry length
site
Residuals
Fry weight
site
Residuals
df
2
60
df
2
60
df
2
60
df
2
60
Sum of Sq
19.2
367.6
Sum of Sq
37.5
895.1
Sum of Sq
4.6
280.6
Sum of Sq
16.1
73.9
Sum of Sq
546.2
3512.9
Mean Sq
9.58
6.13
Mean Sq
18.74
14.92
Mean Sq
2.32
4.68
Mean Sq
8.04
1.23
Mean Sq
273.10
58.55
F Value
1.5631
F Value
1.2562
F Value
0.4966
F Value
6.5265
F Value
4.6644
Pr(F)
0.2179
Pr(F)
0.2921
Pr(F)
0.6110
Pr(F)
0.0027
Pr(F)
0.0131
Do not distribute, quote or cite
C-46
Draft Document
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Figure 1. Rainbow trout percent normal (100 - % craniofacial deformities) as a
function of the logarithm of selenium concentration in eggs (Exposure Variable).
Untransformed values reported in mg Se/kg tissue wet weight. The curve represents
projections from the fitted logistic equation.
as 40 -
Log(Se in eggs ww)
Figure 2. Rainbow trout percent normal (100 - % skeletal deformities) as a
function of the logarithm of selenium concentration in eggs (Exposure Variable).
Untransformed values were reported in mg Se/kg tissue wet weight. The curve
represents projections from the fitted logistic equation.
.6 .8 1.0 1.2
Log(Se in eggs ww)
Do not distribute, quote or cite
C-47
Draft Document
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Figure 3. Plot of percent abnormal for craniofacial, skeletal and finfold deformities and
edema against selenium concentration in brook trout eggs ww, 2000 and 2001 data.
!=S 40 -
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infold Deformities (°A
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S_
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0 2 4 6 8 10 12 14 16
Egg Se concentration
(|j,g/g, wet weight)
0 2 4 6 8 10 12 14 16
Egg Se Concentration
(|j,g/g, wet weight)
Do not distribute, quote or cite
C-48
Draft Document
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The effect levels determined using the within streams approach resulted in values based on ww in eggs.
The primary tissue for which the reproductive effect levels were based, eggs, was converted from ww to
dw using the average percent moisture of 61.2% for rainbow trout eggs reported by Seilor and Skorupa
(2001).
Chronic Values: Brook trout: between streams approach
No effects at ECi0 level at 7.78 mg Se/kg eggs ww or 20.5 mg Se/kg eggs dw;
egg. Chronic value is >20.5 mg Se/kg eggs dw.
Rainbow trout: Within streams approach
ECio value (skeletal deformities) at 8.2 mg Se/kg egg ww or 21.1 mg Se/kg egg
dw. Chronic value is 21.1 mg Se/kg eggs dw.
Do not distribute, quote or cite C-49 Draft Document
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Kennedy, C.J., L.E. McDonald, R. Loveridge, M.M. Strosher. 2000. The effect of bioaccumulated
selenium on mortalities and deformities in the eggs, larvae, and fry of a wild population of cutthroat trout
(Oncorhynchus clarki lewisi). Arch. Environ. Contam. Toxicol. 39:46-52.
Test Organism:
Exposure Route:
Study Design:
Effects Data :
Cutthroat trout (Oncorhynchus clarki lewisi; spawning adults, 3-6 years)
Dietary and waterborne - field exposure
Total selenium concentrations measured at the time the eggs were taken were
<0.1 |o,g/L from the reference site and 13.3 to 14.5 |o,g/L at the exposed site.
At reference and exposed site (Fording River, BC, Canada which receives
drainage from open-pit coal mining), eggs were stripped from females (n=20
from reference site; n=17 from exposed site) and fertilized from milt from one
male collected at each site. Fertilized eggs were reared in well water and
examined for time to hatch, deformities (craniofacial, finfold, skeletal and yolk
sac malformations), and mortalities. Inspection of deformities in eggs was
performed using 40X magnification.
No significant correlations between the selenium concentrations in the eggs from
either site and: hatching time (reference, 25.5-26.5 days; exposed, 22-25.5 days);
percent deformities preponding (reference, 0-2.4%; exposed, 0-0.34%); percent
deformities after ponding (reference, 0-0.26%; exposed, 0-0.09%); percent
mortalities preponding (reference, 1.5-70.3%; exposed, 1-100%); percent
mortalities after ponding (reference, 0.3-4.3%; exposed, 1.5-43.7%); total percent
mortalities (reference, 2.8-55.8%; exposed, 3.7-100%). The average selenium
residues in tissues were as follows:
Site
Reference
Exposed
Adult fish liver, mg Se/kg
dw
8.2; Range: 3.4-14.6
36.6; Range: 18.3-1 14
Adult fish muscle, mg Se/kg
dw
2.4; 1.4-3.8
12.5; Range: 6.7-41
eggs, mg Se/kg dw
4.6
21.2
Chronic Value:
>21.2 mg Se/kg dw in eggs
>12.5 mg Se/kg dw in muscle
Do not distribute, quote or cite
C-50
Draft Document
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Hardy, R.W. 2005. Effects of dietary selenium on cutthroat trout (Oncorhynchus dark!) growth and
reproductive performance. Report for Montgomery Watson Harza. December 14, 2005.
Test Organism:
Exposure Route:
Test Duration:
Study Design:
Effects Data:
Chronic Value:
Cutthroat trout (Oncorhynchus clarki, 0.9 g)
Dietary only
Six experimental dietary treatments were produced by cold extrusion. The
formulation of the diet was designed to be similar to commercial trout diets and
had a proximate composition of 45% protein and 16% lipid. Seleno-methionine
diluted in distilled water (100 (ig/L) was added in appropriate volumes to each
batch of feed to facilitate pelleting. Measured dietary selenium concentrations
were 1.2 (control), 3.8, 6.4, 9.0, 11.5, and 12 mg Se/kg dw. Fry were fed initially
at a rate of 10 times per day 6 days each week to apparent satiation. Feeding
frequency decreased as fish grew.
124 weeks (865 days, 2.5 yrs)
Groups of 50 fish were placed into triplicate tanks (145 L) receiving 4-15 L/min
of hatchery water at 14.5EC and fed one of the six experimental diets. The fish in
each tank were bulk-weighed and counted every 14 days for the first 12 weeks of
the experiment, and then every 4 weeks until 48 weeks. Samples offish for
whole-body selenium analysis were taken at each sampling date for the first 12
weeks followed by every 3 months thereafter. After six months of feeding, the
fish were transferred to 575 L tanks and the number of replicate tanks per dietary
treatment was reduced to two. After 80 weeks of feeding, the fish were
transferred to 1050 L outdoor tanks each supplied with 70 L/min of constant
temperature (14.5°C) spring (hatchery) water. After 2.5 years of the feeding trial,
fish were spawned and whole body selenium level, egg selenium level, % eyed
eggs, % hatched eggs, and % deformed larvae were examined.
No signs of toxicity (reduced growth or survival relative to controls) were
observed in fish fed the highest dietary selenium treatment (12 mg Se/kg dw)
after the first 80 weeks of exposure just prior to transfer outdoors. No signs of
clinical disease were evident, and no relationship was found between feed
conversion ratios and the level of selenium added to the feed. Average whole
body selenium levels of female Henry's Lake cutthroat trout at spawning at 2.5 to
3 years of age were 5.87, 9.10, 11.37 and 5.61 mg Se/kg dw in the four highest
dietary treatments. Average egg selenium levels in the same four dietary
treatments were 6.61, 5.05, 5.18, and 16.04 mg Se/kg dw. Percent survival from
the eyed stage to hatching varied among treatment groups, with the control and
the highest Se dietary treatment having the second highest survival (85%) and the
fifth dietary treatment group the highest (93%). Percent deformed larvae ranged
from a low of 5.6% in controls to a high of 20.2% in the 6.4 mg Se/kg dw dietary
treatment group; larvae in the two highest dietary treatment groups only
exhibited 7 and 6.8 %, respectively.
The chronic value for embryo/larval deformity is aNOAEC of >11.3 7 mg Se/kg
dw whole-body parent tissue and >16.04 mg Se/kg dw egg.
Do not distribute, quote or cite
C-51
Draft Document
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Rudolph, B-L, I. Andreller, CJ. Kennedy. 2008. Reproductive success, early life stage development,
and survival of Westslope cutthroat trout (Oncorhynchus clarki lewisi) exposed to elevated selenium in an
area of active coal mining. Environ. Sci. Technol. 42: 3109-3114.
Test Organism:
Exposure Route:
Test duration:
Study Design:
Effects Data:
Westslope cutthroat trout (Oncorhynchus clarki lewisi)
Field collected.
In June, 2005, eggs were collected from 12 females from Clode Pond (exposed
site) and 16 females from O'Rourke Lake (reference site). Milt was obtained
from 3-5 males at each site. Clode Pond is on the property of Fording River Coal
Operations in Southeast British Columbia with reported selenium concentrations
of 93 (ig/L. O'Rourke Lake is an isolated water body into which Westslope
cutthroat trout were stocked in 1985, 1989 and 1992 and has selenium levels
reported <1 (ig/L.
Through the end of yolk sac absorption (at swim-up) by the alevins.
Individual batches of eggs were fertilized in the field with 2 ml composites of
milt. Water-hardened eggs were transported to the rearing laboratory. Eggs and
alevins were monitored daily for fertilization, hatching and mortality. After the
yolk sacs were absorbed, alevins were sacrificed and preserved in Davidson's
solution.
All viable fry (n = 4,922) after yolk absorption were observed for the frequency
and severity of skeletal (lordosis, kyphosis, and scoliosis), craniofacial (head,
eyes or jaw), and fin malformations as well as edema. The authors used a
graduated severity index (GSI) for deformities in which fry were scored 0
(normal) to 3 (severe) based on the level of defect.
Eggs with the four highest Se concentrations (86.3 to 140 mg/kg dw) collected
from Clode Pond fish died before reaching the laboratory (Table 1). Excluding
the eggs that died from females CP1, CP3, CP4 and CP5, fertilization (total eggs
reaching the eyed stage/total eggs x 100) was not related to Se concentrations in
the eggs (Table 1). The percent of alevins (post hatch to swim-up stage) that died
was related to the selenium concentration in the eggs; the ECi0 estimated by
TRAP is 24.1 mg Se/kg dw Figure 1). Note: The data used in the TRAP analysis
excluded the variable from OL1. These are data from the reference lake in which
only 57% of the larvae survived. Alevin survival was meaningfully higher in the
other 15 clutches of eggs from the reference site (87.3 to 99.8%). Analysis of
combined egg and alevin survival resulted in a similar ECio estimate. The
selenium in muscle data was not amenable for analysis with alevin survival using
TRAP. ECio and EC20 estimates for muscle were derived using a least squares
regression of the egg and muscle data reported by Rudolph et al.
[Semuscle]= 4.0853 +0.39 l[Seegg]
(R2 = 0.9094)
Deformity analysis was not performed on the alevins that died prior to the swim-
up stage. Therefore, due either to dead eggs or dead alevins, the occurrence and
severity of deformities were assessed on four clutches of eggs from Clode Pond
(CP2, CP6, CP11 and CP12) with a range of 11.8 to 20.6 :g Se/g dw and 15 of
Do not distribute, quote or cite
C-52
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the 16 clutches (all eggs died in OL8) from O'Rourke Lake (Table 1). There was
no correlation between egg Se concentration and frequency of deformity or
edema. Statistical differences between sites were observed (p < 0.05) for skeletal
deformities and edema for both the frequency of the occurrence and the severity
score (Table 2). Note: the percent and severity score of skeletal deformities were
greater in the reference site than in the exposed site.
The effect level for this study was based on the alevin mortality data and not the
deformity measurements. Although edema occurred statistically more often at the
exposed site (87.7% at Clode Pond, 61.2% at O'Rourke Lake), it was not
correlated with selenium levels in the eggs. Also the greater occurrence of
skeletal malformations in the reference site confounded the use of statistical
differences between sites to determine effect levels for this study.
Effect Concentration: 24.11 mg Se/kg dw in eggs; 13.51 mg Se/kg dw in muscle
Table 1. Fertilization, egg mortality and alevin mortality for offspring from individual fish
collected in Clode Pond and O'Rourke Lake.
Fish ID
Clode Pond
(exposed site)
CP1
CP2
CP3
CP4
CP5
CP6
CP7
CP8
CP9
CP10
CPU
CP12
avg
SD
Muscle [Se]
mg/kg dw
38.8
11.8
40.4
46.1
50.4
34.7
39
7
35.4
35.5
11.3
13.4
30.3
15.1
Egg [Se] Dead eggs, Dead alevins,
mg/kg dw Fertilization, % % %
88.3
16.1
86.3
121
140
51
65.3
11.8
46.8
75.4
16.9
20.6
61.6
42.4
0
99.7
0
0
0
99
97.2
73.7
91.3
88.2
79.2
98.6
61
45
100
1.8
100
100
100
7.4
8.9
36.1
36.6
17.6
22.1
3
44
42
NA
0.9
NA
NA
NA
92.6
91.1
0.8
63.2
82.4
1.3
5.1
42
44
O'Rourke Lake
(reference site)
OL1
OL2
OL3
OL4
OL5
OL6
OL7
OL8
OL9
OLIO
8.28
7.7
8.16
8.03
8.12
6.61
8.52
7.22
7.25
7.64
12.9
13.9
12.5
15
14.9
15.2
12.9
12.3
16.7
13.1
100
93.1
99.4
98.2
89.3
76
99.4
30.5
96.4
99.1
28.6
53.1
3.9
14.5
19.3
32
2.1
100
12.8
2.5
42.9
6.9
2.4
12.7
5.3
4
0.2
NA
4.5
5.5
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C-53
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Fish ID
OL11
OL12
OL13
OL14
OL15
OL16
avg
SD
Muscle [Se]
mg/kg dw
8.74
8.2
7.86
8.5
7.62
8.13
7.9
0.6
Egg [Se]
mg/kg dw Fertilization, %
Dead eggs, Dead alevins,
15.6
13.9
15.1
13.1
12.3
12.7
13.9
1.4
96.2
99.1
92.6
79.5
92.4
71
88
18
10.8
16.4
25.9
22.2
11.8
45.2
25
25
2.4
3
2.8
0.5
2.6
4.8
7
10
Table 2. Deformity results (frequency and severity) for offspring from O'Rourke Lake and Clode
Pond. Values are presented as mean ± SE. * indicates a significant difference (p < 0.05)
between means from the two sites.
Frequency of deformity, %
Skeletal*
Craniofacial
Pinfold
Edema*
Severity of deformity, score
Skeletal*
Craniofacial
Pinfold
Edema*
O'Rourke Lake
37.4 ±3.6
10.2 ±2.0
10.6 ±3.1
61.2 ±4.9
0.47 ±0.07
0.12 ±0.03
0.15 ±0.05
0.61 ±0.05
Clode Pond
16.5 ±2.2
5.7 ± 1.0
7.5 ±3.84
87.7 ±2.0
0.18 ±0.02
0.06 ±0.01
0.09 ±0.05
0.88 ±0.02
Do not distribute, quote or cite
C-54
Draft Document
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Figure 1. Survival of Westslope cutthroat trout alevin as a logistic function of the logarithm of the
selenium concentration in eggs.
110
100
90
80
£
. 70
15
> 60
I 5°
j> 40
<
30
20
10
0
1.1 1.2 1.3 1.4 1.5 1.6
Log(Egg [Se] pg/g dw)
1.7
1.8
1.9 2.0
TRAP Output
Parameter summary
Parameter
Log X50
S
YO
Initial Est.
1.35
3.0
96.35
Final Est.
1.5885
2.663
97.59
Std Error
0.0345
0.694
2.17
95% LCL
1.516
1.209
93.04
95% UCL
1.661
4.116
102.13
Effect concentration summary
p
50
20
10
5
1
Xp estimate
38.77
28.73
24.11
20.51
14.36
95% LCL
32.83
20.95
16.03
12.51
7.22
95% UCL
45.78
39.39
36.26
33.63
28.53
Do not distribute, quote or cite
C-55
Draft Document
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Nautilus Environmental. 2011. Evaluation of the Effects of Selenium on Early Life Stage Development
of Westslope Cutthroat Trout from the Elk Valley, BC. Report to Elk Valley Selenium Task Force,
November 24, 2011.
Test Organism:
Exposure Route:
Test Duration:
Study Design:
Effects Data:
Westslope cutthroat trout (Oncorhynchus clarki lewisi)
Field collected. Adult fish were collected and spawned from lentic and lotic
environments in areas proximate to Teck Coal's Fording River Operations. Eggs
were also obtained from fish collected from Connor Lake, a lake located within
the Elk valley watershed not exposed to mine discharges and considered a
reference site and a methodological control.
Fertilized eggs were reared in the laboratory until they reached swim-up fry
stage. A subset of fry surviving at swim-up were reared for an additional 28 days.
Gametes were stripped from the ripe adults in the field during June and July 2008
and transported immediately to the laboratory in coolers containing wet ice. Eggs
were fertilized in the laboratory. After stripping the eggs, female fish were
sacrificed and the whole body stored on ice for later Se analysis. For a given
female, approximately 240 fertilized eggs were divided into four replicates of 60
eggs. In cases when fewer eggs were available three replicates of 60 eggs were
used. If less than 180 eggs were available, either 3 or 4 replicates of 30 were
used. Females with less than 90 eggs were not used. The fertilized eggs were
maintained in the laboratory until the fry reached swim-up at which point
deformities were assessed. Survival was also assessed up to swim-up. In test
chambers in which there were at least 40 surviving fish at swim-up, one-half of
the surviving fish were maintained for an additional 28 days. Survival, length,
weight and deformities were assessed in the 28-day post swim-up test.
The number, type and severity of deformities were measured at swim-up and at
the end of the 28-day post swim-up test. Deformity assessments were conducted
on recently killed fresh fish to avoid artifacts caused by preservation. A
graduated severity index (GSI) was assigned to each of four types of
deformity/abnormality: skeletal, craniofacial, finfold and edema. Graduated
Severity Index (GSI) methods followed those described in Holm et al. (2003) and
Rudolph et al (2006; 2008).
Survival of the larvae from hatch through swim-up spawned from the four fish
collected from the reference site, Connor Lake, ranged from 73 to 92% (egg Se
4.32 to 7.31 mg/kg dw) (Table 1). Larval survival at swim-up was also generally
high for fish collected in the Se exposed sites up to egg Se concentration 29.6
mg/kg dw (Table 1, Figure 1). Larvae exposed above this egg Se concentration
had poor to no survival. Larvae from one fish (POOS 11) below this threshold did
have poor survival (11.7%). The authors noted that the many of the eggs from
this fish displayed an unusual distribution of lipid vesicles which resulted in
greater than 50% mortality in the first 24 hours due to egg breakage. The
remaining eggs may have been compromised due to the organic material released
during the egg breakage.
The rate of deformities in larvae at swim-up showed no relationship with Se in
egg through 29.6 mg/kg dw (Table 2).
Do not distribute, quote or cite
C-56
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The results of the 28-day post swim-up test showed no relationships between
larval survival or deformities and egg Se (Table 3). The authors also measured
the length and weight of larvae at the end of the 28 day test; neither of which
showed a relationship with egg Se concentration.
Se Tissue Concentrations. Two analytical laboratories (A and B) measured Se in
the eggs. The mean difference in egg Se concentrations between the two
laboratories was 34.2%. To better understand the difference between the two
laboratories, five egg samples (i.e., from five different fish) from this study were
sent to both laboratories in 2010. Both laboratories digested the eggs using the
methods they used in their own 2008 original analysis. The respective digestates
were split and then shared between laboratories. Both labs then measured
selenium in their own digestates and the digestate received from the other lab.
The results of this follow-up study showed that when each lab used their own
digestion procedures Laboratory A had on average 43% higher measurements in
the 2008 analysis and 23% higher in the follow-up 2010 analysis. When each lab
measured selenium using the same digestate the difference in the Se
measurements between labs was on average only 1 to 8%. The authors concluded
that although both laboratories employed acceptable and approved practices,
Laboratory A used a more efficient digestion process resulting in higher Se
measurements. To compensate for the reduced Se measurements in Laboratory B,
its values were increased by 34.2%. The measurements made by Laboratory A
are marked in Table 1; unmarked values are Laboratory B measurements
increased by 34.2%.
Effect Concentration: The most sensitive endpoint determined by TRAP was larval survival at swim-
up. TRAP was used to model larval survival with the entire egg Se dataset that
included egg Se measurements from Laboratory A and adjusted measurements
from Laboratory B (EC10 = 26.6 mg/kg egg dw; Figure 1) and using only the egg
Se measurements from Laboratory A (Figure 2). Because the Laboratory A
dataset estimated slightly lower EC values, the EC10 of 24.02 mg/kg egg dw is
the selected effect concentration for this study.
Do not distribute, quote or cite C-57 Draft Document
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Table 1. Summary of westslope cutthroat trout larvae surviving to swim-up per parent female (fish
ID) including location of collection of parent female and concentration of selenium in the eggs.
Proportion surviving
Fish ID
YO93
CL1
R082
CL4
CL2
CL3
P00815
R026
P00823
R039
R086
R077
R042
R055
R043
R074
POOS 11
P00809
P00803
R078
GO99
O087
O085
WO52
R069
R071
WO94
UT101
Se egg,
Location mg/kg dw Replicates
Lentic
Reference
Lotic
Reference
Reference
Reference
Lotic
Lotic
Lotic
Lotic
Lotic
Lotic
Lotic
Lotic
Lotic
Lotic
Lotic
Lotic
Lotic
Lotic
Lotic
Lentic
Lentic
Lentic
Lotic
Lotic
Lentic
Lentic
3.88*
4.32
5.21
5.96*
6.82
7.31
7.6
12.53
12.71
12.9
13.4*
14.29
16.44
16.5
16.85
17.8*
19.25
19.72
24.8*
29.61
34.2*
54.7*
56.8*
61.1*
65.61
72.9
73.1
74.67
4
4
3
4
4
4
3
4
4
4
4
3
3
4
4
4
1
4
4
4
4
4
4
4
4
4
4
4
Replicate
mean
0.8125
0.9167
0.9056
0.7333
0.8333
0.8542
0.8222
0.5792
0.8875
0.6042
0.9417
0.6444
0.8
0.8792
0.8667
0.9375
0.1167
0.7667
0.9375
0.8825
0.2083
0.07083
0
0
0
0
0
0
Replicate Replicate Number Total
min max survivors number
0.6667
0.8833
0.8333
0.6
0.7
0.8167
0.7167
0.5
0.85
0.55
0.85
0.6167
0.7
0.7833
0.7667
0.8833
0.1167
0.65
0.9333
0.8333
0.1667
0.01667
0
0
0
0
0
0
0.9167
1
0.95
0.8
0.9167
0.8833
0.95
0.65
0.95
0.65
0.9833
0.6667
0.9
0.9667
0.9667
0.9833
0.1167
0.8833
0.95
0.9333
0.2667
0.2
0
0
0
0
0
0
195
220
163
176
200
205
148
139
213
145
226
116
72
211
104
225
7
184
225
105
50
17
0
0
0
0
0
0
240
240
180
240
240
240
180
240
240
240
240
180
90
240
120
240
60
240
240
119
240
240
240
240
240
240
240
240
*Laboratory A dataset
Do not distribute, quote or cite
C-58
Draft Document
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Table 2. Summary of westslope cutthroat trout larval deformities to swim-up per parent female
(fish ID) including location of collection of parent female and concentration of selenium in the eggs.
Fish ID
YO93
CL1
R082
CL4
CL2
CL3
P00815
R026
P00823
R039
R086
R077
R042
R055
R043
R074
P00809
P00803
GO92
R078
GO99
Location
Lentic
Lentic
Lotic
Lentic
Lentic
Lentic
Lotic
Lotic
Lotic
Lotic
Lotic
Lotic
Lotic
Lotic
Lotic
Lotic
Lotic
Lotic
Lotic
Lotic
Lotic
Se egg, Skeletal Craniofacial Finfold Edema Deformities
mg/kg dw combined combined combined combined combined
3.88*
4.32
5.21
5.96*
6.82
7.31
7.6
12.53
12.71
12.9
13.4*
14.29
16.44
16.5
16.85
17.8*
19.72
24.8*
26.1
29.61
34.2*
4.5%
7.6%
1.2%
4.3%
11.1%
5.0%
0.0%
2.1%
1.9%
2.1%
2.7%
1.7%
1.2%
0.0%
0.9%
2.7%
3.9%
2.7%
0.0%
1.8%
14.5%
0.9%
1.9%
1.3%
7.3%
3.7%
2.0%
2.7%
2.1%
2.9%
1.9%
1.0%
10.4%
0.0%
2.8%
2.6%
1.8%
2.8%
0.9%
1.9%
0.0%
53.9%
4.4%
1.0%
2.5%
1.7%
0.8%
1.0%
0.0%
0.7%
1.8%
2.9%
0.0%
0.9%
0.0%
1.0%
1.8%
0.9%
3.3%
0.0%
1.9%
1.0%
6.8%
1.9%
1.0%
0.0%
0.7%
3.0%
0.0%
2.9%
1.4%
5.6%
4.9%
0.0%
12.2%
2.6%
2.9%
1.7%
0.9%
4.7%
0.9%
4.4%
2.9%
28.2%
7.7%
9.5%
3.7%
12.6%
15.9%
7.0%
5.6%
2.1%
7.4%
9.9%
2.7%
15.5%
2.6%
4.7%
4.4%
3.6%
9.0%
4.5%
4.4%
5.7%
64.7%
*Laboratory A dataset
Do not distribute, quote or cite
C-59
Draft Document
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Table 3. Summary of larval survival and rates deformities after the 28-day post swim-up test per
parent female (fish ID) including location of collection of parent female and concentration of
selenium in the eggs.
Fish ID
CL1
CL2
CL3
CL4
Y093
R082
P00815
P00823
R086
R077
R055
R074
P00809
P00803
Location
Reference
Reference
Reference
Reference
Lentic
Lotic
Lotic
Lotic
Lotic
Lotic
Lotic
Lotic
Lotic
Lotic
Sample
size (n)
112
93
96
68
93
71
69
105
112
36
101
106
65
108
EggSe
(mg/kg dw)
4.3
6.8
7.3
6
3.9
5.2
7.6
12.7
13.4
14.3
16.5
17.8
19.7
24.8
Survival
(%)
99.1
99
91.7
98.6
95.6
87.4
91.1
96.3
97.2
92.4
95.9
93.1
91.7
95.7
Skeletal
(%)
0
0
0
0
0
0
0
0
0
2.8
0
0
0
0
Craniofacial
(%)
0
0
1
0
0
2.9
1.2
0
0.9
2.8
4.6
0
0
0
Pinfold
(%)
0
0
1
4.3
2
0
1.4
0
0
2.8
0
0
0
1
Total
(%)
0
0
2
4.3
2
2.9
2
0
0.9
4.2
4.6
0
0
1
Do not distribute, quote or cite
C-60
Draft Document
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1.1
1.0
.9
-g
3
0
O
Q.
O
ol
.7
.6
.5
.4
.3
.2
.1
n
\
* \
\
\
• \
i i i i i i
.4
.6
1.0 1.2 1.4
Log(Se egg), mg/kg dw
1.6
Figure 1. TRAP 1.20 Analysis type - Tolerance distribution; Model option - Triangular distribution
(3 parameters). Includes Laboratories A and B datasets.
Xp Estimates
p
50
20
10
5
0
Xp Estimate
35.992
29.449
26.616
24.779
20.850
95% LCL
34.831
28.128
25.229
23.356
19.376
95% UCL
37.192
30.832
28.080
26.289
22.437
Do not distribute, quote or cite
C-61
Draft Document
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1.1
1.0
.9
1
1
0
o
Q.
0
ol
.7
.6
.5
.4
.3
.2
.1
n
\
\
\
• \
i i i i i i
.4
.6
1.0 1.2 1.4
Log(Se egg), mg/kg dw
1.6
Figure 2. TRAP 1.20 Analysis type - Tolerance distribution; Model option - Triangular distribution
(3 parameter). Includes Laboratory "A" dataset only. *
Xp Estimates
p
50
20
10
5
0
Xp Estimate
33.75
26.92
24.02
22.16
18.237
95% LCL
32.56
25.54
22.55
20.65
16.686
95% UCL
34.99
28.37
25.57
23.77
19.933
* Although some scientists have attempted to explain certain occurrences of improved response with
increasing concentration in terms of nutrient selenium sufficiency-deficiency, the concentrations involved
in this study are too high to for selenium deficiency to be an explanation. The figure's apparent bi-phasic
measured response is thus best explained as being a chance outcome of noise.
Do not distribute, quote or cite
C-62
Draft Document
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Formation Environmental. 2012. Appendix E - Yellowstone Cutthroat Trout Adult Laboratory
Reproduction Studies. Technical Support Document: Proposed Site-Specific Selenium Criterion, Sage
and Crow Creeks, Idaho. Prepared for J.R. Simplot Company. January 2012.
Test Organism:
Exposure Route:
Test Duration:
Study Design:
Effects Data:
Yellowstone cutthroat trout (Oncorhynchus clarkii bouvieri)
Field collected. Adult female and male Yellowstone cutthroat trout were
collected at five field sites from four streams near the Smokey Canyon mine.
addition Yellowstone cutthroat trout eggs were obtained from a hatchery as
method controls.
In
Test duration was from hatch through 15 days post swim up, and averaged 55-56
days for larvae hatched from field collected fish and 64 days for larvae hatched
from laboratory collected fish.
Eggs were collected from 15 ripe females at five sites from four streams
upstream and downstream of the Smokey Canyon mine. This included one
selenium impacted stream downstream of the mine, Sage Creek (LSV), one site
along Crow Creek upstream of Sage Creek (CC-150) and one site along Crow
Creek downstream of Sage Creek (CC-350), and in sites within the reference
streams Deer Creek (DC), and South Fork Tincup Creek (SFTC). Eggs were
fertilized in the field with milt collected from males collected at the same site as
females. Fertilized eggs were water hardened at the site using stream water, then
placed in oxygenated plastic bags and stored on ice in the dark (cooler) for
transportation to laboratory. In addition, eggs were collected from 16 ripe
females obtained from Henry's Lake hatchery (HL) to serve as method controls.
Hatchery females were stripped of eggs and fertilized by milt from males
obtained from the same hatchery. For field and hatchery fish, Se was measured
in adult fish (whole body) and in eggs of field collected females.
A target of approximately 600 fertilized eggs from each female (were placed in
egg cups for hatching and monitoring. After swim up, remaining fry were thinned
to a target of 100 fry/treatment and monitored for an additional 15 day post swim
up feeding trial.
Endpoints measured in the laboratory were hatch, survival (hatch to swim up, and
hatch through 15 days post swim up), and deformities. Deformities were
combined as assessed as having at least one deformity, or being fully free of
deformities (i.e., normal).
Eggs failed to hatch for one of the field treatments (SFTC-1), and six of the
hatchery treatments, resulting in a final dataset of eggs fertilized from 14 field
collected fish and 10 hatchery fish. Se concentrations in eggs obtained from field
J OO
collected females ranged from 11.4 mg/kg in Deer Creek through 47.6 mg/kg in
Crow Creek downstream of Little Sage Creek (Table 1). Se concentrations in
eggs obtained from Henry's Lake hatchery fish ranged from 0.83 mg/kg - 3.23
mg/kg (Table 1). Se concentrations in whole body tissue samples obtained from
field collected females ranged from 8.17 mg/kg in Deer Creek through 25.7
mg/kg in Crow Creek downstream of Little Sage Creek (Table 1). Se
concentrations in whole body tissue samples obtained from Henry's Lake
hatchery fish ranged from 0.23-0.91 mg/kg (Table 1).
Do not distribute, quote or cite
C-63
Draft Document
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Three endpoints (proportion free from deformities, proportion surviving from
hatch through 15 day post swim up, and proportion surviving from hatch through
15 days post swim up AND free of deformities were analyzed. Selenium
concentrations and respective counts are shown in Table 1. For all tables, each
sample I.D. represents eggs hatched from a single female fish. Plots of each
endpoint are shown in Figure 1. All analyses were performed in TRAP (version
1.22) using tolerance distribution analysis and assuming a triangular data
distribution.
TRAP analysis was performed using count data provided to EPA by Formation
Environmental. In the information provided by Formation Environmental, the
combined endpoint (survived and free from deformities) was calculated as the
number of normal larvae divided by the sum of larvae assessed for deformities
and larvae that died during the test (Table 1). The above calculation implies that
the number of larvae assessed for deformities is equivalent to the number of
larvae that survived the test. However, it is noted that in at least one treatment
(LSV2C-001), no larvae survived through swim up (Formation Environmental
2012), meaning that all deformity assessments for that treatment had to have
been performed on dead fish. For these analyses, both % survival and % fully
normal + survived were set to 0% for that treatment. It is not clear whether dead
larvae were assessed in other treatments based on the available information, so
for this analysis, no other treatments were adjusted. For each endpoint, the ECio
was calculated and the goodness of fit was assessed using R2, which for a
nonlinear regression model is calculated as (1-(residual sum of squares/total sum
of squares)).
Rates of total deformities were high and variable across the selenium
concentration range (Figure 1). The ECio for total deformities was 9.769 mg/kg
egg dw, and the R2 = -0.03, meaning that a horizontal line through the average y-
value would have a better fit than the fitted model. The ECio for survival was
25.25 mg/kg, and the R2 = 0.23 (Figure 1). Finally, the ECio for the combined
endpoint (survived the test and fully free from deformities was 11.78 mg/kg, with
an R2=0.02. Of the three endpoints, the ECio of 25.25 mg/kg is selected because
it is not confounded by high variability among low concentration treatments and
provides the best model fit of the three endpoints.
Effect Concentration: 25.25 mg Se/kg dw in eggs
Do not distribute, quote or cite C-64 Draft Document
-------
Table 1. Yellowstone cutthroat trout selenium concentrations, survival, and deformity data
from hatch to test end.
Sample IDa
CC-150/001
CC-350/001
CC-350/002
CC-350/003
CC-350/004
CC-350/005
DC/001
DC/002
DC/003
DC/004
HL/002
HL/003
HL/004
HL/006
HL/007
HL/008
HL/011
HL/012
HL/013
HL/015
LSV2C/001
LSV2C/002
LSV2C/003
LSV2C/004
EggSe
mg/kg
17.6
27.9
29.7
22.3
14.6
47.6
22
15.4
11.4
12.7
2.03
2.48
1.36
0.83
2.26
1.87
3.23
1.58
1.93
2.06
40.1
30.0
35.6
30.5
WBbSe
mg/kg
16.3
20.7
19.4
17.0
16.7
25.7
8.17
9.07
8.63
16.6
0.45
0.44
0.36
0.36
0.44
0.28
0.31
0.23
0.72
0.91
19.4
21.0
18.6
22.5
#Free
From
Deformities
22
14
143
73
149
91
95
133
59
7
5
121
154
21
120
147
69
112
148
0
2
40
92
107
# Assessed
For
Deformities
182
138
602
330
480
392
275
465
380
38
39
302
416
244
404
412
296
454
483
36
200
319
487
476
#Died
33
120
83
36
19
71
30
26
39
23
10
19
20
103
18
37
22
27
24
6
536
105
138
75
# Survived
182
138
602
330
480
392
275
465
380
38
39
302
416
244
404
412
296
454
483
36
0
319
487
476
# Assessed +
#Died
215
258
685
366
499
463
305
491
419
61
49
321
436
347
422
449
318
481
507
42
536C
424
625
551
a - CC - Crow Creek; DC - Deer Creek; LSV2C - Sage Creek; HL - Henry's Lake (Hatchery)
b - whole body
c - does not include the 200 fish assessed that were dead prior to assessment, as all fish for that treatment
died during the swim up stage in this sample.
Do not distribute, quote or cite
C-65
Draft Document
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1.0 n
in
0
:E 0.8 -
Q 0.6 -
o
0
£
LL
Q.
2
Q-
0.4 -
0.2 -
o.o
0.0
1.0 n
T3
C
0.8
ro
X^
0
>
|
W
cL
o
0.4
0.2
0.5 1.0 1.5
log (mg Se/kg egg dw)
U.U I I I I W
0.0 0.5 1.0 1.5
log (mg Se/kg egg dw)
1.0 -i
"ro
E 0.8
o
~z_
"g 0.6
ro
0
>
'£ 0.4
3
W
Q.
0 0.2
Q.
n n
0 ™
* • \ *
* * ~~—~«^» •
m , *
0.0 0.5 1.0
log (mg Se/kg egg dw)
1.5
Figure 1. Concentration response relationships of Yellowstone cutthroat trout deformities
(top), survival (middle), and deformities+survival (bottom) in response to selenium
concentrations in eggs. ECios (mg Se/kg egg dw) and R2 as follows: deformities (ECio=
9.769, R2= -0.03); survival (ECio= 25.25, R2= 0.23); deformities+survival (ECio= 11.78, R2=
0.02).
Do not distribute, quote or cite
C-66
Draft Document
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Colder Associates. 2009. Development of a Site-specific Selenium Toxicity Threshold for Dolly Varden
Char. Report to Northgate Minerals Corporation, PO Box 3519, Smithers, British Columbia. Report
Number 04-1421-101/2000.
Test Organism:
Exposure Route:
Test duration:
Study Design:
Effects Data:
Dolly Varden (Salvelnius malmd)
Field collected.
Adult Dolly Varden char were collected from reference (North Kemess Creek),
high Se exposure (Upper Waste Rock Ponds and Creek) and moderate Se
exposure (lower Waste Rock Creek) sites during September 22 to 24, 2008. Eggs
were stripped from females and fertilized with milt from males collected from the
reference site. Fertilized eggs were taken to the laboratory for testing.
The test was terminated when 90% of the larvae reached swim-up, approximately
5 months after fertilization.
Approximately 30 fertilized eggs were added to each replicate rearing container.
The number of replicates per female parent ranged from one to four depending
on the number of eggs available. Embryos were maintained in 4 L containers
with 3.5 L dechlorinated tap water in a static-renewal system (3 renewals
times/week) at 5°C. The condition of the embyos and alevins were observed
daily and any dead individuals were counted and removed. Test termination
occurred over a 3-day period during February 11 to 13, 2009. The hatched larvae
were sacrificed using an overdose of the anesthetic, clove oil. Individual length
and weight were measured on each fry, and deformity analysis was performed on
fresh unpreserved larval fish using 40X magnification.
A graduated severity index (GSI) was used for deformity assessment (skeletal,
craniofacial, and finfold as well as edema). The narrative criteria were the same
as used by Holm et al. (2005) and Rudolph et al. (2008).
Alevin survival was not related to Se concentration in the eggs (Table 1). Almost
all of the mortality occurred during the egg stage. Only 4 alevins died during the
study, 1 from Fish #19 and 3 from Fish #2, both females collected at an exposed
site. The prevalence of deformities increased sharply after the selenium egg
concentration exceeded 50 mg/kg dw (Table 1, Figure 1). The proportion of
Dolly Varden larvae with any type of deformity (skeletal, craniofacial, and
finfold as well as edema) as a function of the log of the selenium concentration in
the eggs using TRAP (logistic equation) produced an ECio value of 56.22 mg/kg
dw eggs (Figure 1).
Do not distribute, quote or cite
C-67
Draft Document
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Table 1. Selenium concentration in the eggs of Dolly Varden char and the survival of alevins to the
swim-up stage and the proportion of larvae without any type of deformity.
Fish
#
1
2
5
6
15
19
9
12
17
Sample
ID
WRC-
F105
WRC-F61
WRC-
F103
WRC-F83
WRC-
F104
WRC-F86
NK-F30
NK-F29
NK-F21
Location
Waste Rock Creek
Waste Rock Creek
Waste Rock Creek
Waste Rock Creek
Waste Rock Creek
Waste Rock Creek
North Kemess Creek
North Kemess Creek
North Kemess Creek
[Se]
eggs
mg/kg
dw
56.6
65.8
32.6
51.9
56.3
60.5
11
10.5
5.4
Survival of eggs to
up
Initial
120
120
29
120
60
120
30
46
90
End
71
81
29
115
48
115
1
15
86
swim-
%
59
68
100
96
80
96
3
33
96
Proportion of
larvae
without any
type of
deformity
0.89
0.58
0.97
0.97
0.90
0.72
a
1.00
0.91
SCD1 Redd#l
SCD2 Redd #2
Southern Collection
Ditch 10.3 30 18 60
Southern Collection
Ditch 24.7 40 32 80
1.00
1.00
Do not distribute, quote or cite
C-68
Draft Document
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Figure 1. Proportion of Dolly Varden alevin without any type of deformity as a logistic function of
the logarithm of the selenium concentration in eggs (TRAP).
1.2r
o
o
03
03
O
C
O
o
Q.
O
.6
r=0.933
1.0 1.2 1.4 1.6
Iog10 Egg Se (mg/kg dw)
1.8
2.0
Guess
Final
SE 95% LCL 95% UCL
LogXSO
Slope
YO
Total
Model
Error
1.844
4.152
0.975
ECx
50
20
10
5
1
DF
9
2
7
1.829
6.963
0.980
EC
67.42
60.12
56.22
52.85
46.11
SS
1.74E-01
1.63E-01
1.11E-02
0.007
1.252
0.017
95% LCL
64.92
57.96
53.00
48.64
40.13
MS
1.93E-02
8.13E-02
1.58E-03
1.812
4.003
0.939
95% UCL
70.01
62.35
59.64
57.43
52.99
F
51.429
1.845
9.924
1.021
P
0.99993
Do not distribute, quote or cite
C-69
Draft Document
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AECOM. 2012. Reproductive success study with brown trout (Salmo truttd). Data quality assurance
report. Final. December 2012.
Formation Environmental. 2011. Brown Trout Laboratory Reproduction Studies Conducted in Support
of Development of a Site-Specific Selenium Criterion. Prepared for J.R. Simplot Company by Formation
Environmental. Revised October 2011.
Test Organism:
Exposure Route:
Test duration:
Study Design:
Brown trout (Salmo truttd)
Field collected.
Adult female and male brown trout were collected at three field sites from two
streams downstream of the Smokey Canyon mine. In addition, brown trout eggs
were obtained from two hatcheries as method controls.
Embryo-larval monitoring to 15 days post swim-up.
Eggs were collected from 26 ripe female brown trout at three field sites
downstream of the Smokey Canyon mine. These included one site on the highly
impacted Sage Creek (LSV2C) as well as two sites along Crow Creek (CC-150
and CC-350) downstream of the conflux with Sage Creek. The downstream -
most station along Crow Creek (CC-150) was intended to be a field control. Eggs
were fertilized in the field with milt collected from males collected at the same
site as females. Fertilized eggs were water hardened at the site using stream
water, then placed in oxygenated plastic bags and stored on ice in the dark
(cooler) for transportation to laboratory. Se was measured in adult fish (whole
body) and in eggs of field collected females. In addition, eggs were collected
from 8 ripe females obtained from the Saratoga National Fish Hatchery (SC) to
serve as method controls. Similar to field-caught fish, SC hatchery females were
stripped of eggs and fertilized by milt from males obtained from the same
hatchery. As a result of lower than expected hatch rates and fungal contamination
in some SC hatchery samples, additional hatchery fish were obtained (as already
fertilized eyed embryos) from the Spring Creek Trout Hatchery (SPC), which
were divided into four treatments.
Approximately 600 fertilized eggs from each female (or 600 eyed embryos for
SPC treatments) were placed in egg cups for hatching and monitoring. After
swim up, remaining fry were thinned to a target of 100 fry/treatment and
monitored for an additional 15 day post swim up feeding trial. Test termination
ranged from 83 to 88 days after hatch for all but the Spring Creek Hatchery egg
treatments, which occurred 50 days after the arrival of fertilized, eyed embryos
from that hatchery.
Endpoints measured in the laboratory study were fecundity, hatch, growth,
survival/mortality, and feeding success (growth) post swim up. Larval brown
trout were also evaluated for deformities (craniofacial, vertebral, fin) and edema.
For this study, deformities were combined and assessed as having at least one
deformity, or being fully free of deformities (i.e., normal).
Do not distribute, quote or cite
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Effects Data: Se concentrations in eggs collected from 26 ripe females at 3 field locations
ranged from 6.2-12.8 mg Se/kg dw at CC150, 6.9-14.0 mg Se/kg dw at CC350,
and 11.2-40.3 mg Se/kg dw at LSV2C. Se concentrations in hatchery eggs ranged
from 0.76-1.2 mg Se/kg dw at the SC hatchery, and were 0.73 mg Se/kg dw at
the SPC hatchery. The Se whole body concentration in field collected fish ranged
from 7.2-22.6 mg/kg dw at LSV03, 4.7-8.4 mg/kg dw at CC150, and 5.5-9.2
mg/kg dw at CC350. Se whole body concentrations in SC hatchery fish ranged
from 2.5-4.3 mg/kg dw. Hatchery data were combined with field data and
included in all analyses.
Because of concerns raised in a U.S. Fish and Wildlife (2012) review of the
Formation Environmental (2011) report regarding the consequences offish lost
due to an overflow event resulting from a drain that became clogged with food
during the 15 day post swim up portion of the test, all endpoints were analyzed
using a both an "optimistic" and a "worst-case" scenario. The U.S.FWS (2012)
review of the Formation study proposed all treatments that lost fish to the
overflow event should be excluded from the ECio calculation, because they were
more likely to have been dead or deformity. As an alternative to that proposal,
the "worst-case" scenarios were introduced here to examine that hypothesis, by
treating all fish lost to overflow as either dead or deformed, rather than excluding
those treatments altogether. In the "optimistic" scenario, the overflow event was
treated as a random technician error unrelated to selenium toxicity, and any lost
fish were removed from the calculation. In other words, fish lost to overflow
were assumed to be equally likely to have been dead or deformed to fish that
were not lost.
A second assumption for these analyses was also based on a comment made in a
U.S. FWS (2012) review of the Formation Environmental report, where it was
noted that fish that survived but failed to reach swim up would likely have died
in the wild. This occurred among the offspring of the five females with the
highest egg selenium concentrations, ranging from 26.8-40.3 mg Se/kg egg
(LSV2C-003, -004, -005, -010, and -021). For all endpoints that were analyzed
from hatch through the end of the 15 day post swim up feeding trial, all fish that
failed to reach swim up were assumed to be dead, with respect to survival.
Three endpoints: percentage fully free from deformities (% normal), percentage
surviving (% survival), and percentage surviving AND fully free of deformities
(% alive and normal)) were analyzed. All analyses were performed on hatch
through 15 days post swim up, with the exception of the % survival endpoint (see
below), which was also analyzed from hatch to swim up. All analysis was
performed in TRAP (version 1.22) using tolerance distribution analysis and
assuming a triangular data distribution.
Combined Survival and Deformity Endpoint
Selenium concentrations and counts of total larvae and fully normal larvae (alive
and normal) are included in Table 1. The ECio for the worst case scenario was
20.65 mg/kg and the ECio for the optimistic scenario was 21.16 mg/kg (Figure
1). Although the combined endpoint is theoretically the most sensitive endpoint,
because it combines the effects of mortality and deformities, the combined
endpoint did not yield the lowest ECi0s for this study. Because of the particular
data distribution, the percentage number of alive and normal larvae decreased
Do not distribute, quote or cite C-71 Draft Document
-------
from approximately 56% and 60% in the 20.5 mg/kg and 17.7 mg/kg treatment
concentrations in the worst case scenario (60% and 74%, respectively, in the
optimistic scenario) to 0% in the 26.8 mg/kg and higher treatment concentrations
(Figure 1). At selenium concentrations 20.5 mg/kg and below, the percentage of
larvae that were alive and normal were highly variable, even among the low
selenium hatchery treatments. The abrupt decline in alive and normal larvae
between the 20.5 mg/kg and 26.8 mg/kg treatment concentrations resulted in an
ECio that was slightly greater than 20.5 mg/kg for both scenarios.
Do not distribute, quote or cite C-72 Draft Document
-------
Table 1. Brown trout selenium concentrations and survival + deformity data (combined endpoint) from hatch to test end (15 days post
swim up). Worst case counts assumed that all fish lost to the overflow event during the post swim up portion of the study would have been
deformed.
Sample IDa
SC-001
SC-002
SC-003
SC-004
SC-005
SC-006
SC-007
SC-008
SPC-001C
SPC-002C
SPC-005C
SPC-006C
CC-1 50-009
CC-150-011
CC-150-012
CC-150-013
CC-150-015
CC-150-016
CC-150-017
CC-150-018
CC-1 50-020
CC-350-006
CC-350-007
CC-350-008
LSV2C-002
LSV2C-003
LSV2C-004
Whole
body
Se, mg/kg
dw
3.6
4.1
3.7
4.3
3
3.1
2.7
2.5
8.4
5.6
6.7
5.9
6
7
5.6
4.7
7.2
9.2
5.5
8.5
8.9
13.8
17.9
EggSe
mg/kg
dw
0.76
0.94
0.83
0.92
1.2
1.2
1
0.96
0.73
0.73
0.73
0.73
12.8
8.4
8.5
8.4
9.1
7.5
6.6
6.9
6.2
14
6.9
9.5
12.8
40.3
36
# Normal
that were
# dead at
Normal assessment
63
72
131
46
23
457
93
283
427
371
400
427
106
87
156
137
210
13
99
195
453
120
68
269
483
2 2
16 16
#
Normal
and
alive
63
72
131
46
23
457
93
283
427
371
400
427
106
87
156
137
210
13
99
195
453
120
68
269
483
0
0
# Live fish
assessed for
deformities
115
113
302
140
42
535
137
359
570
545
561
556
142
266
282
310
445
23
163
486
558
386
131
338
544
0
0
# Fish lost to
overflow
# Fish died during post
during test swim up test
8
4
7 9
28
6
8
30
6 10
8
20
8
17
11
2
12
46 26
14
3 43
7 33
16
6
26
10 20
21 28
4 16
395
289
# Live fish
assessed + # died
during test.
"Optimistic case"
123
117
309
168
48
543
167
365
578
565
569
573
153
268
294
356
459
26
170
502
564
412
141
359
548
395
289
# Live fish
assessed + # died
during test + #
lost during post
swim up. "Worst
case"
123
117
318
168
48
543
167
375
578
565
569
573
153
268
294
382
459
69
203
502
564
412
161
387
564
395
289
Do not distribute, quote or cite
C-73
Draft Document
-------
Sample IDa
LSV2C-005
LSV2C-008
LSV2C-010
LSV2C-012
LSV2C-016
LSV2C-017
LSV2C-019
LSV2C-020
LSV2C-021
Whole
body
Se, mg/kg
dw
13.6
9.6
22.6
7.2
9.2
13.2
8.6
11.3
20
EggSe
mg/kg
dw
26.8
17.7
38.8
13.2
13.4
20.5
12.5
11.2
28.1
#
Normal
8
147
5
217
440
110
267
240
8
# Normal
that were
dead at
assessment
8
5
8
#
Normal
and
alive
0
147
0
217
440
110
267
240
0
# Live fish
assessed for
deformities
0
194
0
554
530
150
390
296
0
# Fish died
during test
267
4
97
17
20
28
22
5
404
# Fish lost to
overflow
during post
swim up test
45
19
39
36
# Live fish
assessed + # died
during test.
"Optimistic case"
267
198
97
571
550
178
412
301
404
# Live fish
assessed + # died
during test + #
lost during post
swim up. "Worst
case"
267
243
97
571
550
197
451
337
404
a SC - Saratoga National Fish Hatchery; SPC - Spring Creek Trout Hatchery; CC - Crow Creek; LSV - Sage Creek
b Test end was 15 days after swim up.
0 Arrived as fertilized, eyed-eggs. No whole body Se measurement possible.
Do not distribute, quote or cite
C-74
Draft Document
-------
1.0 -i
a> 0.8
CO
T3
C
(0
(/)
0)
0.6 -
E
£ 0.4
Q
M—
O
£ 0.2 -
0.0
0.0
0.5 1.0
log (mg Se/kg egg dw)
1.0 -,
£ 0.8 -
£
CO
T3
C
(0
0.6 -
•^
E
£ 0.4 -
0)
Q
M—
O
0)
£ 0.2 -
0.0
0.0
0.5 1.0
log (mg Se/kg egg dw)
Figure 1. ECi0s for combined (mortality + deformity) endpoint. Top - worst case scenario.
ECio=20.65 mg/kg. Bottom - optimistic scenario. ECi0=21.16 mg/kg.
Do not distribute, quote or cite
C-75
Draft Document
-------
Deformity Endpoint
Selenium concentrations, counts of larvae assessed for deformities, and counts of
normal larvae are included in Table 2. In the selenium draft document submitted
to external peer review, the ECi0 for the worst case deformity endpoint was
calculated as 15.91 mg/kg (Figure 2), and the ECi0 for the optimistic deformity
endpoint was calculated as 18.36 mg/kg (Figure 3). The ECio of 15.91 mg/kg for
the worst case scenario deformity endpoint was presented as the final ECi0 for
the brown trout SMCV, because it was the lowest (most conservative) of all
measured endpoints. During the public comment phase, it was noted that the
ECioS for deformity calculated by TRAP are dependent on the initial model
conditions, most notably the standard deviation parameter, which is the
parameter for the falling limb of a tolerance distribution model.
The effects of initial model conditions for the resulting ECIO of the deformity
endpoint- worst case scenario are shown in Figure 2. In this figure, the initial
values of the logXCSO (the EC50 of the model fit) and the y-intercept are
identical (logXC50=1.4, y-intercept=0.6), but the initial values of the slope
parameter are varied. When the initial standard deviation term is set at 0.064 or
higher, the final standard deviation is solved to be approximately 0.125, and the
model converges at or near an ECio of 15.91 (Figure 2 - top). When the initial
standard deviation term is set at 0.063 or lower, the final standard deviation is
solved to be approximately 0.0378, and the model converges at or near an ECi0
of 21.58 (Figure 2 - bottom).
When multiple minima are present, the most statistically appropriate model, or
global minima, is the model where the residual sum of squares is minimized.
The residual sum of squares for the ECi0=15.91 mg/kg model is 1.152, which is
larger than the residual sum of squares of 1.064 for the ECi0=21.58 mg/kg model,
meaning the model with the ECi0=21.58 mg/kg provides the best fit to the data.
Within the range where the models diverge, at concentrations of 17.7 mg/kg and
higher, the difference between the residual sum of squares is even more
pronounced, as the error in the 21.58 mg/kg ECi0 model is 5.3-fold less than the
error in the 15.91 mg/kg ECio model (0.0198 versus 0.105 sum of squares). As a
final line of evidence in support of the ECio = 21.58 mg/kg for the worst case
deformity endpoint, when counts are converted to proportions, and the nonlinear
regression model (threshold sigmoid model shape) is run in TRAP, the resulting
ECio is 22.1 mg/kg.
The effects of initial model conditions for the resulting ECIO of the deformity
endpoint- optimistic scenario are shown in Figure 3. In this figure, the initial
values of the logXC50 (the EC50 of the model fit) and the y-intercept are identical
(logXC50=1.3, y-intercept=0.62), but the initial values of the slope parameter are
varied. When the initial standard deviation term is set at either 0.056-0.078, or
0.117 or higher, the final standard deviation is solved to be approximately 0.122,
and the model converges at or near an ECio of 16.36 (Figure 3 - top). When the
initial standard deviation term is set from 0.079 through 0.116, the final standard
deviation is solved to be approximately 0.101, and the model converges at or
near an ECio of 18.37 (Figure 2 - middle). Finally, when the initial standard
deviation term is set at 0.055 or lower, the final standard deviation is solved to be
approximately 0.0349, and the model converges at or near an ECio of 21.94
(Figure 3 - bottom). The corresponding residual sums of squares for the three
Do not distribute, quote or cite C-76 Draft Document
-------
model fits are as follows: EC10=16.36 mg/kg (1.065), EC10=18.36 mg/kg (1.039),
ECio=21.95 mg/kg (0.9326). As discussed regarding the worst case scenario for
the deformity endpoint, ECio of 21.95 mg/kg is the model with the lowest
residual sum of squares, and is the recommended ECi0 for the optimistic scenario
deformity endpoints because it provides the best fit to the data. As a final line of
supporting evidence, when counts are converted to proportions, and the nonlinear
regression model (threshold sigmoid model shape) is run in TRAP, the resulting
ECio is 22.1 mg/kg.
One of the reasons for the multiple minima of the deformity endpoint models is
the high variability in deformities at concentrations at or below 20.5 mg/kg
(Figures 2-3). Even when additional variability is not introduced, by assuming
fry lost to the overflow event were deformed (the optimistic scenario), variability
in deformity rates is high (Figure 3). In addition to the high variability in
deformities among the field samples and the SC hatchery samples (the hatchery
samples processed by Formation Environmental), there appears to be a site effect
in deformity rates as well. As can be seen in Figures 2 and 3, deformity rates
among field samples appear to be greater for fish hatched from eggs collected in
the two Crow Creek sites (CC-150, CC-350) compared to Sage Creek (LSV-2C).
Although the difference in means is not statistically significant at the 95%
confidence level, the confidence bounds separate at around the 20% confidence
level, suggesting that this might not be a random effect. This pattern holds for all
but the five highest concentrations (26.8-40.3 mg/kg), all from Sage Creek,
which clearly fall above the threshold for selenium toxicity. If the result of
higher deformities among Crow Creek sites is not a random artifact, it suggests a
confounding factor, unrelated to selenium exposure. Whether the higher
deformity rates represent random variation, population differences, other
environmental quality differences (unrelated to Se), or methodological issues is
unclear.
Do not distribute, quote or cite C-77 Draft Document
-------
Table 2. Brown trout selenium concentrations and deformity data from hatch to test end (15 days
post swim up). Worst case counts assumed that all fish lost to the overflow event during the post
swim up portion of the study would have been deformed.
Sample
IDa
SC-001
SC-002
SC-003
SC-004
SC-005
SC-006
SC-007
SC-008
spc-oor
SPC-002C
SPC-005C
SPC-006C
CC-150-
009
CC-150-
011
CC-150-
012
CC-150-
013
CC-150-
015
CC-150-
016
CC-150-
017
CC-150-
018
CC-150-
020
CC-350-
006
CC-350-
007
CC-350-
008
LSV2C-
002
LSV2C-
003
LSV2C-
004
LSV2C-
Whole
body
Se, mg/kg
dw
3.6
4.1
3.7
4.3
3
3.1
2.7
2.5
8.4
5.6
6.7
5.9
6
7
5.6
4.7
7.2
9.2
5.5
8.5
8.9
13.8
17.9
13.6
EggSe
mg/kg
dw
0.76
0.94
0.83
0.92
1.2
1.2
1
0.96
0.73
0.73
0.73
0.73
12.8
8.4
8.5
8.4
9.1
7.5
6.6
6.9
6.2
14
6.9
9.5
12.8
40.3
36
26.8
#
Normal
63
72
131
46
23
457
93
283
427
371
400
427
106
87
156
137
210
13
99
195
453
120
68
269
483
2
16
8
# Assessed for
deformities.
"Optimistic
Case"
115
113
302
140
42
535
137
359
570
545
561
556
142
266
282
310
445
23
163
486
558
386
131
338
544
100
142
149
# Lost to # Assessed for
overflow deformities plus
during post # lost. "Worst
swim up test Case"
115
113
9 311
140
42
535
137
10 369
570
545
561
556
142
266
282
26 336
445
43 66
33 196
486
558
386
20 151
28 366
16 560
100
142
149
Do not distribute, quote or cite
C-78
Draft Document
-------
Sample
IDa
005
LSV2C-
008
LSV2C-
010
LSV2C-
012
LSV2C-
016
LSV2C-
017
LSV2C-
019
LSV2C-
020
LSV2C-
021
Whole
body
Se, mg/kg
dw
9.6
22.6
7.2
9.2
13.2
8.6
11.3
20
EggSe
mg/kg
dw
17.7
38.8
13.2
13.4
20.5
12.5
11.2
28.1
#
Normal
147
5
217
440
110
267
240
8
# Assessed for
deformities.
"Optimistic
Case"
194
80
554
530
150
390
296
172
# Lost to
overflow
during post
swim up test
45
19
39
36
# Assessed for
deformities plus
# lost. "Worst
Case"
239
80
554
530
169
429
332
172
a SC - Saratoga National Fish Hatchery; SPC - Spring Creek Trout Hatchery; CC - Crow Creek; LSV -
Sage Creek
b Test end was 15 days after swim up.
0 Arrived as fertilized, eyed-eggs. No whole body Se measurement possible.
Do not distribute, quote or cite
C-79
Draft Document
-------
1 .U
CD
W
CD
¥. 0.8-
"GO
L
O
:>
, 0.6-
CD
E
| 0.4-
c
o
o 0.2 -
o
CL
o.o -J
• SC Hatchery
A T SPC Hatchery
* m A • CC-150
w « • CC-350
V * A " A LSV-2C
T •
_ A
* =v A
• • • \
" • \
• • • A \
• • A \
\
• \
EC10=15.91 mg/kg \A
AI \A
a
tiii
0.0 0.5 1.0 1.5 2
log(Dg Se/kg dw)
CD
CD
O
"GO
L
O
^
i
"CD
E
o
c
0
o
Q.
0
ol
i .\j
0.8 -
0.6 -
0.4 -
0.2 -
n n -
• A
w A
v v A
T • .
• • A
A A^~i
• \
" • \
• • • A \
• • A 1
^T 1
• \
EC10=21. 58 mg/kg \
L
• SC Hatchery
V SPC Hatchery
• CC-150
+ CC-350
A LSV-2C
A
L, A
^•v^ A
o.o
0.5 1.0
log(Dg Se/kg dw)
1.5
2.0
Figure 2. Comparison of EClOs for deformities-worst case scenario as a function of initial
conditions. When initial standard deviations are set to 0.064 or higher, the ECi0 for worst case
deformities converges at or near 15.91 mg/kg (top). When initial standard deviations are set to
0.063 or lower, the ECio for worst case deformities converges at or near 21.58 mg/kg (top). The
residual sum of squares is lower for the ECi0=21.58 mg/kg model compared to the ECi0=15.91
mg/kg model (1.064 vs. 1.152), indicating that the ECi0=21.58 mg/kg model provides a better fit to
the data.
Do not distribute, quote or cite
C-80
Draft Document
-------
o
^
••s.
o
1
"ro
0
c
o
o
CL
O
ol
.0
'S.
o
ro
o
c
0
1
0
Q_
0.8 -
0.4 -
0.2 -
On
• SC Hatchery
0 T SPC Hatchery
9 • ^A • CC-150
¥ « AA + CC-350
T • A A LSV-2C
oDD \
* a A \
EC10=1 6.36 mg/kg V
0.0 0.5 1.0 1.5 2.0
log (Dg Se/kg dw)
1 n
0.8 -
0.6 -
0.4 -
0.2 -
n n -
• SC Hatchery
0 V SPC Hatchery
£ • ^A • CC-150
¥ • AA • CC-350
T • A A LSV-2C
V ^ S^
• °cP \
D A \
EC10=18.37 mg/kg V
0.0 0.5 1.0 1.5 2.0
log (Dg Se/kg dw)
o
E
••s.
o
i
"ro
E
0
z
c
o
t
o
CL
O
ol
1 .U
0.8 -
OC
.D
0.4 -
0.2 -
n n
• A
J • A
T 0 A
• • ~\
• SC Hatchery
T SPC Hatchery
• CC-150
• CC-350
A LSV-2C
• • •• 1
* \
• J \
« A \
- _ \
• « * \
\
EC10=21. 94 mg/kg \ A
Ai A
ij^
^^^^ A
0.0
0.5
1.0
1.5
2.0
log (Dg Se/kg dw)
Figure 3. Comparison of ECi0s for deformities-optimistic scenario as a function of initial conditions.
Depending on initial parameter values, the final model converges on three ECi0s. Residual sums of
squares for the three models, top to bottom are (1.065,1.039, and 0.9326). The lowest residual sum
of squares is for the ECi0 =21.95 mg/kg model, indicating that it provides a better fit to the data
than the other two models.
Do not distribute, quote or cite
C-81
Draft Document
-------
Survival Endpoint
Selenium concentrations and estimated counts of larvae surviving from hatch
through 15 days post swim up are included in Table 3. Estimated counts are used
for calculating survival through the 15 day post swim up test because larvae were
thinned to a target of 100 individuals/treatment prior to the onset of the 15 day
post swim up test, and final survival is calculated as the product of survival from
hatch to swim up and survival during the 15 day post swim up test. The ECi0 for
the worst case survival scenario was 16.78 mg/kg and the ECi0 for optimistic
survival scenario was 20.40 mg/kg (Figure 4). The ECi0s for both models were
stable across a wide range of initial conditions, and are not subject to the multiple
minima issue of the deformity endpoint. The ECi0 of 16.78 mg/kg for the
optimistic is effectively identical to the ECio for the worst case survival scenario
of 16.76 mg/kg presented in the peer-reviewed response to the FWS review of
the Formation Environmental study (Taulbee et al. 2012).
However, in light of the high survival in all hatchery and field samples through
20.5 mg/kg, concern has been expressed that the worst case scenario is
particularly unrealistic for the survival endpoint. Because measured survival was
high, the effects of the overflow event (worst case scenario) were more
pronounced, as the difference in ECi0s among the worst case and optimistic
scenarios (20.40 mg/kg) was greatest for this endpoint.
In addition, the principle scientist of the brown trout study stated in the public
comments to the selenium draft document submitted for external peer review:
"escaped fry were observed swimming in the water bath where the treatment
containers were being held. These fry congregated near the treatment cells. Dead
or dying fish were not observed." This observation was not included in the
original Formation Environmental (2011) report, where fry lost to overflow were
treated in a manner consistent with the "optimistic" scenario.
Do not distribute, quote or cite C-82 Draft Document
-------
Table 3. Brown trout selenium concentrations and survival data from hatch to test end (15 days post swim up). Worst case counts
assumed that all fish lost to the overflow event during the post swim up portion of the study would have been deformed.
Sample IDa
SC-001
SC-002
SC-003
SC-004
SC-005
SC-006
SC-007
SC-008
SPC-001C
SPC-002C
SPC-003C
SPC-004C
SPC-005C
SPC-006C
CC-1 50-009
CC-150-011
CC-150-012
CC-150-013
CC-150-015
CC-150-016
CC-150-017
CC-150-018
CC-1 50-020
CC-350-006
CC-350-007
CC-350-008
LSV2C-002
LSV2C-003
LSV2C-004
LSV2C-005
Whole
body
Se, mg/kg
dw
3.6
4.1
3.7
4.3
3
3.1
2.7
2.5
8.4
5.6
6.7
5.9
6
7
5.6
4.7
7.2
9.2
5.5
8.5
8.9
13.8
17.9
13.6
EggSe
mg/kg
dw
0.76
0.94
0.83
0.92
1.2
1.2
1
0.96
0.73
0.73
0.73
0.73
0.73
0.73
12.8
8.4
8.5
8.4
9.1
7.5
6.6
6.9
6.2
14
6.9
9.5
12.8
40.3
36
26.8
#Eggs
Hatche
d
144
138
340
189
70
564
188
396
598
20
585
21
589
593
173
288
314
402
479
89
223
522
584
432
181
407
584
404
309
287
Prop.
Survival
. Hatch
to swim
up
0.951
0.978
0.982
0.868
0.914
0.988
0.856
0.985
0.987
1.000
0.966
1.000
0.986
0.971
0.942
0.993
0.965
0.891
0.971
0.966
0.969
0.969
0.990
0.944
0.950
0.951
0.993
0.079
0.414
0.387
Prop
survival. Post
swim up.
"Optimistic
Case"
0.990
0.990
0.989
0.971
1.000
0.990
0.970
1.000
1.000
1.000
1.000
1.000
1.000
1.000
0.990
1.000
0.990
0.973
1.000
1.000
1.000
1.000
1.000
0.980
0.988
0.986
1.000
0.281
0.477
0.622
Prop
survival. Post
swim up.
"Worst
Case"
0.990
0.990
0.890
0.971
0.984
0.990
0.970
0.900
1.000
1.000
1.000
1.000
1.000
1.000
0.990
1.000
0.990
0.720
1.000
0.500
0.670
1.000
1.000
0.980
0.790
0.710
0.840
0.281
0.477
0.622
Prop survival.
Hatch to end.
"Optimistic
case"
0.942
0.968
0.971
0.842
0.914
0.978
0.830
0.985
0.987
1.000
0.966
1.000
0.986
0.971
0.933
0.993
0.955
0.866
0.971
0.966
0.969
0.969
0.990
0.926
0.938
0.938
0.993
0.022
0.197
0.240
Prop survival.
Hatch to end.
"Worst case"
0.942
0.968
0.874
0.842
0.900
0.978
0.830
0.886
0.987
1.000
0.966
1.000
0.986
0.971
0.933
0.993
0.955
0.641
0.971
0.483
0.649
0.969
0.990
0.926
0.751
0.675
0.834
0.022
0.197
0.240
Est.#
survived.
Hatch to end.
"Optimistic
case"
136
134
330
159
64
551
156
390
590
20
565
21
581
576
161
286
300
348
465
86
216
506
578
400
170
382
580
9d
61d
69d
Est.#
survived.
Hatch to end.
"Worst case"
136
134
297
159
63
551
156
351
590
20
565
21
581
576
161
286
300
258
465
43
145
506
578
400
136
275
487
9d
61d
69d
Do not distribute, quote or cite
C-83
Draft Document
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Sample IDa
LSV2C-008
LSV2C-010
LSV2C-012
LSV2C-016
LSV2C-017
LSV2C-019
LSV2C-020
LSV2C-021
Whole
body
Se, mg/kg
dw
9.6
22.6
7.2
9.2
13.2
8.6
11.3
20
EggSe
mg/kg
dw
17.7
38.8
13.2
13.4
20.5
12.5
11.2
28.1
#Eggs
Hatche
d
263
108
591
570
217
471
357
424
Prop.
Survival
. Hatch
to swim
up
0.989
0.231
0.971
0.965
0.885
0.953
0.986
0.288
Prop
survival. Post
swim up.
"Optimistic
Case"
0.982
0.440
1.000
1.000
0.963
1.000
1.000
0.730
Prop
survival. Post
swim up.
"Worst
Case"
0.540
0.440
1.000
1.000
0.780
0.610
0.640
0.730
Prop survival.
Hatch to end.
"Optimistic
case"
0.971
0.102
0.971
0.965
0.852
0.953
0.986
0.210
Prop survival.
Hatch to end.
"Worst case"
0.534
0.102
0.971
0.965
0.690
0.582
0.631
0.210
Est.#
survived.
Hatch to end.
"Optimistic
case"
255
lld
574
550
185
449
352
89d
Est.#
survived.
Hatch to end.
"Worst case"
140
lld
574
550
150
274
225
89d
a SC - Saratoga National Fish Hatchery; SPC - Spring Creek Trout Hatchery; CC - Crow Creek; LSV - Sage Creek
b Test end was 15 days after swim up.
0 Arrived as fertilized, eyed-eggs. No whole body Se measurement possible.
d Survived but failed to reach swim up. Assumed dead in all hatch to 15 day post swim up analyses.
Do not distribute, quote or cite
C-84
Draft Document
-------
w
CD
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CO
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1.0 T
0.8 -
0.6 -
0.4 -
0.2 -
0.0 -
I • • ua Si A
• "^ B^ • SC Hatchery
• • V T SPC Hatchery
^* A\ • CC-150
* \ 4 CC-350
+ \A A LSV-2C
• • A \
A \
A\
• I
I
\
\
\
\
EC10= 16.78 mg/kg \
0.0 0.5 1.0 1.5 2.0
log(Dg Se/kg dw)
1.0 -
0.8 -
0.6 -
0.4 -
0.2 -
0.0 -
4
v^f • 'hi-iQ i A^ A
9 O D<> "Q^ ^ • SC Hatchery
• \ T SPC Hatchery
^* • CC-1 50
^ CC-350
1 A LSV-2C
1
1
1
1
1
EC10= 20.40 mg/kg
]*_ A«
/-\ /-\ /-\ i~ .,4/-\ -<^ *^/^
log(Dg Se/kg dw)
Figure 4. EClOs for larval survival. The ECio for larval survival under the worst case scenario
assumptions was 16.78 mg/kg (top), and the ECi0 for larval survival under optimistic scenario
assumptions was 20.40 mg/kg (bottom). No multiple minima were observed for either model across
a range of initial model parameters.
Do not distribute, quote or cite
C-85
Draft Document
-------
Survival Endpoint - Assessment of Overflow Loss
An assessment was made to determine whether the loss of fish from the overflow
event during the 15 day post swim up portion of the test was related to survival or
Se treatment concentration measured during the first portion of the test. In this
assessment, data were examined from the perspective of whether the overflow
loss of brown trout during the second stage of the test could reflect dead, dying,
or weak organisms.
First, the relationship between larval survival in the first and second stages of the
test (hatch to swim up, 15 days post swim up) were compared for all treatments
where larvae successfully reached the swim up stage (Figure 5). Overall,
survival in the second stage tracks survival in the first stage (r2=0.6), but survival
in the second stage was noticeably higher in than in the first stage.
1.005
si 1
CO
tS 0.995
•o
-------
0.6
HO n r
re O.b
•o
5 0.4
-§ 0.3
_o
I0'2
t
S! o.i
o
o*«»«** •• * • •
0 0.02 0.04 0.06 0.08 0.1 0.12 0.14 0.16
Brown trout mortality during 1st stage
0.6
Si
re
n c.
O.D
1 *
-------
and wild fish, if leaving the aquarium required swimming over the rim, one
might speculate that previous generations of hatchery fish might have developed
a tolerance to remaining in conditions that might seem crowded to wild
organisms. (That is, however, purely speculative.) Otherwise, the difference
between hatchery and wild fish would seem only to reflect a random artifact,
since the Se concentrations at which the wild fish displayed high overflow losses
are low.
0.6
to n c
fB O.D
"O
5 0.4
-o 0.3
I0'2
t
§ 0.1
O
10 15 20 25
Egg Concentration, mg Se/kg dw
Figure 7. Relationship between egg Se concentration and overflow loss
during the second stage of the test. Larvae from treatment levels 26.8 mg/kg
and higher, which failed to swim up, were excluded.
In summary, the positive correlation between survival during the hatch to swim
up portion of the test and survival during the 15 day post swim up portion of the
test, combined with the lack of a correlation between mortality during the hatch
to swim up portion of the test and overflow loss during the second stage of the
test, suggests that the overflow loss may represent a random technician error not
related to the health of the individuals lost. The relationship between selenium
egg concentrations and overflow loss was lower for the larvae hatched from
hatchery fish compared to the larvae hatched from field collected fish; however,
among field treatments ranging from 6.0-20.5 mg/kg there was no correlations,
further supporting the hypothesis that the overflow event was a random
occurrence unrelated to the health of larval fish.
Survival Endpoint - ECin for the first portion of the test
Because larval survival was measured at the end of the first portion of the test
(hatch to swim up), an alternative approach to measuring survival would be to
calculate the brown trout EC10 for survival for only the first portion of the test.
Selenium concentrations and counts of total larvae and larvae that survived the
first portion of the test are included in Table 4. The hatch to swim up portion of
the test was much longer than the second portion (88 days on average compared
Do not distribute, quote or cite C-88 Draft Document
-------
to 15 days), and more importantly, it avoids the experimental confound
introduced by the loss offish during the overflow event. With this approach, the
second portion of the test would be rejected as inconclusive due to the laboratory
accident.
In contrast to survival endpoints measured from hatch through 15 days post swim
up, survival for all treatments, including larvae from the five treatments of 26.8
mg/kg and higher, where larvae failed to reach swim up, were included.
Although this is theoretically a less conservative approach than the assumption
that larvae that failed to reach the swim up stage would not survive in the wild,
and therefore should be treated as being dead, the inclusion of these larvae results
in a lower ECio (18.09 mg/kg - when non swim up surviving larvae are counted
as survivors; compared to 20.62 mg/kg - when they are assumed to not have
survived in the wild - Figure 8). In contrast to other ECi0 calculations, this
approach is free from all assumptions, and even with respect to larvae that fail to
reach swim up, the ECio calculation is based on measured, rather than assumed,
values.
Unlike survival, deformities could not be analyzed for the first portion of the test
because of a bias introduced during the thinning process prior to the initiation of
the 15 day post swim up portion of the test. During the thinning process, visibly
deformed larvae were selectively removed, so that the fish used in the 15 day
post swim up test were less likely to have been deformed. Because of this
selection bias, only survival could be evaluated from hatch to swim up.
Effect Concentration: 18.09 mg Se/kg dw in eggs for larval survival during the hatch to swim up portion
of the test where all surviving larvae were included.
Do not distribute, quote or cite C-89 Draft Document
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Table 4. Brown trout selenium concentrations and survival data from hatch to swim up (first
portion of the test).
Sample IDa
SC-001
SC-002
SC-003
SC-004
SC-005
SC-006
SC-007
SC-008
SPC-001b
SPC-002b
SPC-003b
SPC-004b
SPC-005b
SPC-006b
CC-150-009
CC-150-011
CC-150-012
CC-150-013
CC-150-015
CC-150-016
CC-150-017
CC-150-018
CC-150-020
CC-350-006
CC-350-007
CC-350-008
LSV2C-002
LSV2C-003
LSV2C-004
LSV2C-005
LSV2C-008
LSV2C-010
LSV2C-012
LSV2C-016
LSV2C-017
LSV2C-019
LSV2C-020
LSV2C-021
Whole body
Se, mg/kg dw
3.6
4.1
3.7
4.3
3
3.1
2.7
2.5
8.4
5.6
6.7
5.9
6
7
5.6
4.7
7.2
9.2
5.5
8.5
8.9
13.8
17.9
13.6
9.6
22.6
7.2
9.2
13.2
8.6
11.3
20
EggSe
mg/kg dw
0.76
0.94
0.83
0.92
1.2
1.2
1
0.96
0.73
0.73
0.73
0.73
0.73
0.73
12.8
8.4
8.5
8.4
9.1
7.5
6.6
6.9
6.2
14
6.9
9.5
12.8
40.3
36
26.8
17.7
38.8
13.2
13.4
20.5
12.5
11.2
28.1
# Larvae
Hatched
144
138
340
189
70
564
188
396
598
20
585
21
589
593
173
288
314
402
479
89
223
522
584
432
181
407
584
404
309
287
263
108
591
570
217
471
357
424
# Larvae Survived -
Hatch to Swim Up
137
135
334
164
64
557
161
390
590
20
565
21
581
576
163
286
303
358
465
86
216
506
578
408
172
387
580
32C
128C
111"
260
25C
574
550
192
449
352
122C
a SC - Saratoga National Fish Hatchery; SPC - Spring Creek Trout Hatchery; CC - Crow Creek; LSV
Sage Creek
b Arrived as fertilized, eyed-eggs. No whole body Se measurement possible.
0 Survived, but failed to reach swim up.
Do not distribute, quote or cite
C-90
Draft Document
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1.0 n
0.0
0.5 1.0
log (mg Se/kg egg dw)
1.5
1.0 n
0.8 -
E
CO
.2
o
"ro
I
15
S: 0.4
0.6 -
CO
I
0.2 -
0.0
EC =18.09 mg/kg
o.o
0.5 1.0
log (mg Se/kg egg dw)
1.5
Figure 8. Larval survival, hatch to swim up. Top panel - larvae that survived but did not
reach swim up assumed to be dead. Bottom panel - larvae that survived but did not reach
swim up counted as surviving.
Do not distribute, quote or cite
C-91
Draft Document
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Besser, J.M., W.G. Brumbaugh, D.M. Papoulias, C.D. Ivey, J.L. Kunz, M. Annis, and C.G.
Ingersoll. 2012. Bioaccumulation and toxicity of selenium during a life-cycle exposure with desert
pupfish (Cyprinodon macularius): U.S. Geological Survey Scientific Investigations Report 2012-5033,
30 p. with appendixes.
Test Organism:
Exposure Route:
Test Duration:
Study Design:
Desert pupfish (Cyprinodon macularius)
Dietary and waterborne. Pupfish were fed the oligochate, Lumbriculus
variegatus, which had been grown on a diet of selenized yeast.
180 days life cycle, 21 days Fl larvae, 58 days Fl juveniles and adults.
Desert pupfish (Cyprinodon macularius), a federally-listed endangered species,
were exposed simultaneously to waterborne and dietary selenium at six exposure
levels (controls and five selenium treatments) in a three-phase life cycle exposure
study. Aqueous exposures were prepared using sodium selenate and sodium
selenite salts at an 85%-15% proportion, respectively. Pupfish were fed the
oligochate, Lumbriculus variegatus, daily to satiation (25 to 30% rations based
on wet weights). Prior to being fed to the pupfish, the oligochaetes were exposed
to aqueous selenium and fed selenized yeast at appropriate concentrations to
attain the target dietary tissue concentrations. The measured concentrations in
water, oligochaetes (pupfish diet), and pupfish tissues for the control and five
treatments during the life cycle exposures.
Treatment
Control
Se-1
Se-2
Se-3
Se-4
Se-5
water
HS/L
nd
3.4
6.2
14
26
53
oligochaetes
mg/kg dw
1.6
5.1
7.3
14
24
52
pupfish, mg/kg dw
Fo WB eggs F! WB
0.75 1 1.2
2.5 3 3.4
3.4 4.4 3.7
6.7 8 6.7
12 13 12
24 27 31
The 85-day Phase 1 exposure was initiated with approximately five week old
juvenile pupfish (F0). Phase 1 consisted of two separate groups with one group
(started two weeks prior to the second group) used for determining survival,
growth and whole body selenium concentrations, and the other group used for
survival assessment and to provide adults for the main reproduction exposure.
Both groups in Phase 1 were similarly exposed to all six treatments, with each
treatment having 8 replicates and 10 fish in each replicate.
At the end of the 85-day Phase 1 exposure, the pupfish were reproductively
mature and were used for the Phase 2 exposure, the main reproduction study. A
preliminary reproduction study was conducted with adults from the first exposure
group of F0 pupfish. These fish were divided into two spawning groups and eggs
were collected on four dates during a 9-day period. The main purpose of the
preliminary study was to confirm the reproductive maturity of the pupfish, but
samples of larvae from this study were used for assessment of deformities. The
main reproduction study in Phase 2 was started with adults from the second F0
exposure. These fish were sorted into spawning groups (1 male and 3 females) in
Do not distribute, quote or cite
C-92
Draft Document
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7-L exposure chambers, with eight replicate spawning groups per selenium
treatment. Spawning activity was monitored by removing (and replacing)
spawning substrates from each chamber three times a week (Monday-
Wednesday-Friday). There were 23 egg collection dates during a 60-day period.
All eggs were counted and eggs collected from eight Wednesdays were used for
hatching success, deformities and FI larval and juvenile growth and survival in
the 58-day Phase 3 exposure. Larvae were examined for developmental
endpoints including edema, delayed development, and skeletal, eye, craniofacial,
and fin deformities.
Effects Data: A summary of the endpoints by each treatment level is shown below.
Table 1. Summary of pupfish toxicity endpoints by exposure treatment (average across all
replicates). There were no statistically significant differences across controls and selenium
amendment treatments for any of the endpoints shown here (1-way ANOVA, a=0.05).
Endpoif Control Se-1 Se-2 Se-3 Se-4 Se-5
FO survival, day 28
FO survival, day 56
FO survival, day 85
FO survival, day 150
FO growth, day 28
FO growth, day 56
FO growth, day 85
FO growth, day 150
Fl survival, day 30
Fl survival, day 58
Fl growth, day 30
Fl growth, day 58
total number eggs
% reduction eggs
avg % deformities, main
avg % deformities, preliminary
100
100
100
91
213
535
935
1718
100
100
73
260
6845
NA
5.3
4.4
100
100
100
94
206
526
998
1763
100
100
73
264
6331
8
2.7
8.8
100
100
100
94
204
486
941
1776
100
93
76
286
4143
39
4.9
11.6
100
100
100
94
198
469
934
1755
100
90
78
286
4386
36
2.4
14.3
100
100
100
91
213
509
914
1673
98
95
77
288
3337
51
11.4
10.7
98
100
100
97
203
447
1053
1606
98
88
58
255
5225
24
8.1
21
'Endpoint units: survival, %; growth, mg wet weight; % reduction eggs is relative to the control.
The authors observed no significant differences in pupfish survival or growth
among treatments. The authors hypothesized the lack of statistically significant
acute effects was because the pupfish in this study were near their chronic
toxicity threshold, as suggested by the (non-significant) mean reductions in
growth (7% in F0 day 150) and survival (12% in Fj day 58) in the highest
selenium treatment (Se-5), relative to controls (Table 1).
Do not distribute, quote or cite C-93 Draft Document
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Egg hatching and larval survival in all selenium treatments (not listed in Table 2)
were within 10 percent of control means, and differences among treatments were
not related to selenium exposure. The authors noted that the highest selenium
treatment, Se-5, did have the lowest larval survival (84%) and lowest combined
egg hatching and larval survival (76 percent). The means frequencies of
deformities were higher in the two highest Se treatments (Se-4 and Se-5, Table
1); however % deformities across treatment levels were not statistically
significant (1-way ANOVA, p=0.13; Beckon et al. (2012). However, overall
deformity rates were statistically significantly higher in a preliminary
reproduction than in the main reproduction test. Beckon et al. (2012)
hypothesized that the reason for the difference in deformity rates between the two
tests was related to the time the eggs were collected relative to the time the
respective spawning groups were isolated. Eggs were collected in the preliminary
reproductive study 1-9 days after the spawning groups were isolated, whereas
spawns used to characterize deformities in the main reproduction test were
collected at least 14 days after the onset of spawning. The larvae produced from
the earlier collected eggs may have been exposed to higher selenium
concentrations in the egg. The pattern of a gradual decrease in egg selenium
concentration over time was observed in the life cycle study.
Egg production varied considerably over the 23 collection dates (Table 2 and
Figure 1). Although each of the selenium treatments had a lower total number of
eggs relative to the control, one-way ANOVAs of cumulative egg production did
not indicate significant differences among treatments on either a per-replicate
basis (p=0.34) or on a per-female basis (p=0.20). Similarly, repeated measures
ANOVA indicated no differences between treatments, but the authors indicated
significant differences among sampling dates and significant interactions of
treatment and date. Because of the lower number of eggs in the selenium
treatments and the significance of the interaction of treatment and time, the
authors concluded that pupfish egg production was adversely affected by
elevated selenium exposure and reported significant reductions in egg production
at treatment levels Se-2 through Se-5 (4.4 to 27 mg/kg dw Se in eggs). The
authors recognized that typically larval survival and deformities are the most
sensitive reproductive endpoint for selenium toxicity and not egg production and
suggested more study is needed to confirm the unusual sensitivity of pupfish egg
production to selenium.
Do not distribute, quote or cite C-94 Draft Document
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Table 2. Number of pupfish collected on each sampling date throughout the study, by treatment
level. Values represent the sum of all eggs collected on a given date for a given Se treatment.
Day
Control
Se-1
Se-2
Se-3
Se-4
Se-5
2
4
7
9
11
14
17
21
23
25
28
30
32
35
37
39
42
44
51
53
56
58
60
136
275
307
265
401
417
448
303
287
340
366
130
323
320
236
326
507
251
380
278
199
202
148
112
173
273
252
136
359
456
664
205
308
273
164
304
427
176
151
140
133
359
63
478
329
396
90
123
301
226
424
333
206
404
141
94
103
104
271
81
41
159
55
66
227
38
138
331
187
67
142
283
169
319
246
163
204
143
143
101
52
78
150
113
184
193
152
338
197
195
410
344
122
188
160
271
265
198
145
163
177
150
95
82
75
74
38
113
101
69
305
56
238
143
109
94
162
432
283
380
401
232
400
175
228
181
132
151
223
38
140
140
137
370
188
222
320
196
Do not distribute, quote or cite
C-95
Draft Document
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ra
T3
c
o
-------
Between Subjects
Source
Se treatment
Error
Sum of Sq. df
2,202.6 5
10,543.5 42
Mean Sq. F-rat. p-value
440,5 1.755 0.143
251.0
Within Subjects
Source Sum of Sq. df Mean Sq. F-rat. p-value
Sampling Date 1,867.5 22 84.89 4.973 <0.001
Se Treatment x Sampling Date 2,566.3 110 23.33 1.367 0.010
Error 15,771.8 924 17.07
As with the results reported in Table 7 of Besser et al. (2012), there was no main
effect of Se treatment (note - for purposes of these analyses and associated text,
"Se treatment" is defined as the control plus the 5 treatments that received Se
amendments), but there was a statistically significant (p<0.05) effect of sampling
date and a significant date by Se treatment interaction. Results were qualitatively
similar because the p-values for Se treatment and sampling day were identical in
both analyses, yet the p-values for the day by Se treatment interaction term were
nearly identical.
A statistically significant sampling date effect means that there were significant
differences in overall egg production on different sampling dates. Daily egg
production per female ranged from 2.176 on day 2 to a high of 7.294 on day 11,
and was variable throughout the study. Of greater interest is the statistically
significant day x Se treatment interaction. What this means is, although there was
not an overall significant effect of Se treatment on egg production per female,
there was a significant Se treatment effect (p<0.05) on egg production per female
on at least one of the 23 sampling dates.
Analysis after Removal of Control Replicate Outlier: Repeated measures ANOVA analysis confirmed
the results reported in Besser et al. (2012). However, as shown on Figure 8b of
Besser et al. (2012), one replicate chamber (replicate g) within the control
treatment had only one surviving female pupfish from day 7 through the end of
the test (day 60), and that replicate also had the highest overall egg production
per female of any test chamber. All replicate chambers in all treatments began
with three female pupfish, and the replicate described above was the only one
with only one surviving female. All three females survived the 60 day test in the
majority of the replicate chambers. In order to determine whether the significant
date by Se treatment interaction was an artifact of this one test chamber, data
were reanalyzed after removing this replicate.
One requirement of repeated measures ANOVA is that the model cannot contain
any missing values. An alternative to repeated measures ANOVA when data are
missing, and the most commonly followed procedure under these circumstances,
is to analyze the data using a mixed model. This was the procedure followed
here.
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The results of a fully balanced mixed model (no missing data) should be identical
to repeated measures ANOVA. As an initial check, the full dataset was
reanalyzed as a mixed model. Sample chamber was the random effect parameter,
and Se treatment, sampling date, and Se treatment by sampling date were the
fixed effect parameters. As expected, the F-ratios for the effects of selenium
treatment, sampling date, and the sampling date by Se treatment interaction were
identical. Next, the data were reanalyzed after removing data from control
replicate g from all sampling dates. Results of this analysis are reported in the
table below.
Mixed Model - Fixed
Effect Numerator df Denominator df F-ratio p-Value
Se Treatment 5 902 1.087 0.366
Sampling Date 22 902 6.042 <0.001
Se Treatment x Sampling Date 110 902 1.310 0.023
T
The statistically significant interaction between Se Treatment and Sampling Date
persisted after removal of the potentially anomalous control treatment chamber
with one female pupfish. In other words, even after removing the one potentially
anomalous control replicate, there were still some individual sampling dates
where the effects of Se treatment were statistically significant (p<0.05).
Se Treatment x Sampling Date Interaction: When a significant interaction is
observed in a repeated measures ANOVA, the next recommended step in the
process is to examine each of the repeated measures (sampling dates) separately
to identify those dates where the significant difference in Se treatment level
occurred. When individual dates for the full dataset (including the replicate with
one surviving female) were analyzed separately, there were significant (p<0.05)
effects of Se treatment level on egg production on days 28, 35, 37, 42, and 53 (1-
way ANOVA, df5)42). There were no significant Se treatment effects on the
remaining 18 sampling dates. ANOVA results are summarized in the table
below.
Sampling Date F-ratio p-value
28 2.501 0.045
35 2.704 0.033
37 3.351 0.012
42 4.294 0.003
53 3.352 0.012
Because of the large number of comparisons (23 individual ANOVA models for
each sampling date), an alpha of 0.05 is inappropriate for this particular analysis.
This is because an alpha of p<0.05 means that a statistically significant result will
be observed 5% of the time due to chance alone (Type I error). In order to control
for the increased likelihood of a Type I error when making multiple comparisons,
the alpha level of 0.05 was adjusted using Sidak's correction (Abdi 2007). For 23
comparisons and an alpha of 0.05 for one comparison, the adjusted alpha using
Sidak's correction is as follows:
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1 - (1 - 0,05)23 = 0,0027
After adjusting alpha to account for the 23 separate sampling dates, there were no
sampling dates with a significant Se treatment effect (p<0.0027). As a result, it
was not necessary to perform post hoc means comparisons tests for any of the
individual sampling dates to determine which Se treatment levels were
significantly different from each other.
Each of the 23 sampling dates for the dataset where the replicate chamber from
the control treatment with one surviving female pupfish was excluded were also
analyzed using one-way ANOVA to determine which sampling dates had
significant Se treatment effects. Significant differences among Se treatment
levels at alpha 0.05 are shown in the table below.
Sampling Date F-ratio p-value
35 2.839 0.027
42 3.164 0.017
53 2.549 0.042
After adjusting alpha to account for the 23 separate sampling dates, there were no
sampling dates with a significant Se treatment effect (p<0.0027). As with the full
dataset, it was not necessary to perform post hoc means comparisons tests for any
of the individual sampling dates to determine which Se treatment levels were
significantly different from each other.
Summary of Repeated Measures Analysis: This analysis demonstrated that although there was a
significant Se treatment by sampling date interaction, regardless of whether or
not the control treatment chamber with one female pupfish was excluded,
differences among Se treatment levels were only observed for a small subset of
the 23 sampling dates. Furthermore, after adjusting alpha to account for multiple
comparisons, one-way ANOVA analyses conducted separately for each sampling
date to locate the source of the Se Treatment x Sampling Date interaction
determined that there were no statistically significant differences among Se
treatment levels on any sampling date, precluding the need to perform post hoc
comparison of means tests to identify significant differences among individual Se
treatments.
Combining Effect Metrics Using a Population Model: To improve the certainty of any conclusions to
be made about the sensitivity of pupfish to selenium, it is also worthwhile to
consider the biological (as opposed to statistical) significance of the observations.
But for total egg production, survival, and deformities, the concentration-
response curves did not show a sufficient concentration-related effect to calculate
an EC10. Nevertheless, because Besser et al. (2012) raised the issue of an
interaction of egg production with time, there is a particular concern that there
could be a delay in egg production that would reduce population growth rate,
even while total numbers of eggs were not significantly affected. This question
was evaluated by constructing a population model corresponding to data
available from the test.
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This modeling approach allows for combining and properly weighting effects on
egg production, timing of egg production, and survival. Percent hatch and percent
deformities were also considered in alternate calculations. Because the model is
only intended for combining the lab data into a unified concentration-response
curve, it cannot be interpreted as making real-world population predictions. The
relevant data were taken from spreadsheets Besser et al. (2012b and 2012c),
which were provided by Besser.
The reproduction and larval endpoints spreadsheet, Besser et al. (2012b),
presents egg production at 23 time points. This information thus allows for 23
adult life stages, each assigned its own fecundity. Another page of this
spreadsheet provides larval survival data, thus defining survival of the early life
stage. The juvenile and adult survival spreadsheet, Besser et al. (2012b), defines
a survival rate shared by these life stages.
For each treatment, the data from the test thus provide all the needed input for 25
life stages: (1) an embryo-larval stage with its own daily survival probability
(along with hatching and deformity percentages, when considered in alternative
calculations), (2) anon-reproducing juvenile stage sharing its treatment's daily
survival probability with the adult stages, and (3 - 25) 23 short-duration adult
stages each with its own egg production, but sharing its treatment's daily survival
probability with the treatment's other adult stages. Use of the data is detailed
below.
Egg Production: Egg production at the test's 23 observation time points is from
the spreadsheet Besser et al. (2012b), expressed as eggs per female per day. The
intent of Besser et al. (2012) was for each treatment to have eight replicates, and
each replicate was to have one male and three females. Only replicates matching
that design were used. Early in the test Control Replicate "g" ended up with only
one female, and was therefore not used here. Se-1 Replicate "h" and Se-3
Replicates "d" and "h" had been inadvertently stocked with two males and two
females, and were likewise not used here. Table 3 shows the time course of egg
production incorporated into the population model. For each treatment, model
fecundity, m1, for life stages i = 3 - 25, is the observed egg production divided by
2, in order to provide female eggs per female per day.
Percent Hatch: The spreadsheet Besser et al. (2012b) presents percent hatch for
eggs collected at selected time points. Within each treatment these were
averaged. In selenium reproductive studies percent hatch is often treated as a
noise variable unrelated to selenium exposure. Consequently, the population
growth calculations were run with and without including percent hatch. When
hatch was incorporated into the calculation, daily fecundity was reduced by
multiplying by percent hatch.
Deformities: The Besser et al. (2012b) spreadsheet also provides deformity
counts for the study's preliminary test and for its main test. Only the main test
results were used here. Counts were totaled for each treatment, and a percentage
calculated. Population growth calculations were performed both with and without
consideration of deformity percentage. For simplicity when considered, a worst
case assumption was made that deformed individuals do not contribute to the
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population. Percent deformity was thereby handled in manner parallel to percent
hatch, by multiplying daily fecundity by percent free of deformity.
Table 3. Life stage durations, and observed eggs per female per day at observation time points
for control and selenium treatments, only with replicates having the design three females and one
male. Model fecundity, m, is set at one-half the observed, to yield female eggs per female.
Repro Study
Obser-vation
Day
-
-
2
4
7
9
11
14
17
21
23
25
28
30
32
35
37
39
42
44
51
53
56
58
60
Assigned
Life Stage
Number
1
2
3
4
5
6
7
8
9
10
11
12
13
14
15
16
17
18
19
20
21
22
23
24
25
Total as £ (duration • eg
Life Stage
Duration
35
85
2
2
3
2
2
3
3
4
2
2
3
2
2
3
2
2
3
2
7
2
3
2
2
gs/f/d) =
Observed Eggs/Female/Day
Control
-
-
2.690
5.548
4.333
5.762
8.024
6.540
6.429
3.345
5.786
6.905
4.794
1.881
5.464
4.373
5.631
6.119
7.349
4.798
1.847
6.310
3.183
3.405
3.810
281.6
Se-1
-
-
2.571
4.048
4.302
5.524
3.238
4.905
7.143
7.881
4.643
7.286
4.317
3.881
7.286
7.310
4.417
3.917
2.222
3.274
2.139
1.512
7.317
7.810
8.226
294.3
Se-2
-
-
1.875
2.563
4.181
4.708
8.833
4.625
2.861
4.208
2.938
1.958
1.431
2.167
5.646
1.132
0.927
4.240
1.056
1.719
1.571
0.823
2.076
8.469
4.115
181.9
Se-3
-
-
1.319
2.153
3.185
3.639
4.528
2.296
1.481
1.764
3.806
2.792
1.306
1.403
1.444
2.880
1.556
3.556
2.500
3.194
2.532
5.403
2.491
9.597
6.347
174.7
Se-4
-
-
2.542
3.917
2.222
5.646
5.521
2.750
2.014
1.698
3.688
3.125
1.319
1.708
1.563
1.028
0.792
2.354
1.403
1.438
2.060
1.333
3.528
3.104
2.271
142.0
Se-5
-
-
1.958
3.375
6.000
5.896
7.917
5.569
3.222
4.167
3.646
4.750
2.514
2.750
3.146
3.097
0.792
2.917
1.944
2.854
2.202
3.917
3.083
7.656
4.271
220.1
Larval Survival: The Besser et al. (2012b) spreadsheet also has data for larval
survival after 14 and 21 days for eggs collected at three time points. The fraction
surviving 21 days was used here. For each treatment, the probability of the early
life stage (i=l) surviving each day equals the fraction surviving for 21 days,
raised to the 1/21 power: a\ = aL = (21-d Surv)1721, shown in Table 4.
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Juvenile and Adult Survival: A second spreadsheet, Besser et al. (2012c), has
data on juvenile and adult survival after 30 and 58 days. The fraction surviving
58 days was used (Table 4). Parallel to the handling of larval survival, for each
treatment the juvenile-adult daily survival probability, OJA = (58-d Surv)1758, as
shown in the table. This value applies to life stages i=2-25 (o2 through 1, there would be a slight youthful bias
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within the life stage, such that slightly more than half would be only 1 day into
the life stage and not ready to graduate, and slightly less than half would be in
their second day and ready to graduate. The above function adjusts for that.1
The projected population growth rate for each treatment was calculated as
follows. The 25x25 projection matrix was placed on an Excel spreadsheet. Each
cell in the diagonal was then modified to subtract the eigenvalue, A, which
represents the population growth rate. That is, each cell in the diagonal was
rewritten as cr^l-y,) - A. The determinant of the 25x25 matrix was then calculated
by function MDETERM. To obtain the population growth rate, Excel's Solver
was then tasked with finding a value for/I that yielded a value of zero for the
matrix determinant. In this case,-10~18 < MDETERM < +10"18 was deemed
sufficiently close to zero. Introducing the constraint to look for X values between
1.01 and 1.04 was found helpful for Solver to find the dominant eigenvalue.
When Solver occasionally could not get the determinant within 10"18 of zero,
probably due to a solution oscillation that can occur because the input values yt
are expressed as a function of the solution output/I, digits were removed from
Solver's best estimate for A, to provide a new starting value with which Solver
could complete the solution.
Effects on Projected Population Growth Rates: Table 5 and Figure 2 show the model results. Figures
2-B, -C, and -D are almost indistinguishable from Figure 2-A, because hatch and
deformity rates varied so little across treatments. Although population growth
rates at 4.4 - 27 mg Se/kg are less than at 1 - 3 mg Se/kg, the 6-fold increase in
concentration from 4.4 - 27 mg Se/kg yields no change in response.
Consequently, the results do not suggest a selenium-related effect, and no ECi0
can be calculated. Based on the combined influences of egg production and
timing, and survival (with or without percentage hatch and deformities), pupfish
does not appear to be among the most sensitive species.
1 The formula for y is undefined (0/0) under the condition a=\ and l=\, so it is not obvious from inspection how it behaves. This
function addresses a model artifact that is called numerical dispersion when it occurs in pollutant transport models. It prevents
overoptimistic rates of moving through the life stages, particularly in the 35-day and 85-day larval and juvenile stages, and allows
a 25-stage model of life duration 180 days to yield precisely the same growth rate as a 180-stage (one day per stage) model,
which was also constructed and checked for comparison. However, in this application where absolute growth rates have no
particular meaning and only relative differences between treatments are of interest, the function does not change the overall
perspective.
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Table 5. Model output: daily population growth rates as k (factor increase) and r (=ln k), for
models that account for survival, fecundity and its timing, and optionally also hatch and/or
deformities. Because k is responding to all the treatment parameters included in the model, its
treatment-to-treatment variations do not exactly track the variations in any single input.
Treat-
ment
Control
Se-1
Se-2
Se-3
Se-4
Se-5
Cone
1
3
4.4
8
13
27
Factors included in model:
All account for survival (<7L , OJA ) and fecundity (m) and its timing
•k
1.0337
1.0346
1.0299
1.0285
1.0291
1.0294
r
0.0332
0.0340
0.0294
0.0281
0.0287
0.0290
Hatch
•k
1.0330
1.0338
1.0284
1.0277
1.0283
1.0288
R
0.0324
0.0333
0.0280
0.0273
0.0279
0.0283
deformity
•k
1.0334
1.0344
1.0295
1.0283
1.0283
1.0288
r
0.0328
0.0338
0.0291
0.0279
0.0279
0.0284
hatch &
•k
1.0326
1.0336
1.0281
1.0275
1.0276
1.0281
deform.
r
0.0321
0.0331
0.0277
0.0271
0.0272
0.0277
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1.2
0)
c
O I r
n. -1
0)
o:
=5" 0.8 -
OJ
4_*
3
f 0.6
£
o
Jo 0.4 -
s
| 02
1
]
1.2
>
2 4 8 16 32
Egg concentration (mg Se/kg dw)
1.2
OJ
c
O -I
a. 1 -
4)
^ G n n G
"g °-8
.3.
3 0.6
43 _ . .j population growth
3- eggs/female-d
•*: o 2 21-d larval surv
Ji 58-d juv-adlt surv
OL-, "vt->«-Lv
'n a 1C ft '• T T i
1 2 4 8 16 32
Egg concentration (mg Se/kg dw)
1.2
1 2 4 8 16 32
Egg concentration (mg Se/kg dw)
Figure 2. Abbott-adjusted pupfish response as modeled population growth rate (solid-filled
symbols) and observed eggs per female per day, larval survival, and juvenile and adult
survival (open symbols). Where used in the population model (to modify fecundity),
hatch and deformity are shown as open symbols. Some open-symbol points are
obscured beneath solid-symbol points. (A) Upper left, egg production and survival only,
(B) upper right, adds in influence of percent hatch, (C) lower left, adds in influence of
deformities, and (D) lower right, adds in influence of percent hatch and deformities.
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Isolating the Influence of Timing of Egg Production: By combining survival with egg production and
its timing in the above analysis, the assessment obscures the influence of timing:
the issue that was the main reason for undertaking population modeling in the
first place. The concern is whether selenium exposure could delay reproduction,
thereby yielding reduced population growth. To help isolate the influence on the
timing of egg production, two population model runs were performed where all
treatments were assigned one of two daily survival rates (0.99 or 0.999) spanning
the full range of daily survival rates observed in the 21 and 58 day survival
calculations. That is, with survival held constant, the only factors varying across
treatments were egg production and timing.
The results are shown in the table below. The Abbott-adjusted results are plotted
in Figure 3. Although the relative differences in Figure 3 population growth rates
are subdued compared to the wider variation in egg production, this is merely a
consequence of the predicted population growth rate being more responsive to
survival than to reproduction. It is still apparent that the variations in total egg
production are affecting growth rate. The question to be addressed here is
whether increasing selenium concentration yields a decline in growth rate beyond
the pattern reflecting total egg production.
Population growth rates, as influenced only by differences in egg
production and timing
Treat-
ment
Control
Se-1
Se-2
Se-3
Se-4
Se-5
Cone
1
3
4.4
8
13
27
With only egg production (m) and its timing
variable across treatments
(7=0.999
•k
.0339
.0338
.0310
.0293
.0293
.0324
r
0.0334
0.0333
0.0306
0.0289
0.0288
0.0318
(7=0.99
•k
1.0246
1.0245
1.0217
1.0201
1.0200
1.0231
r
0.0243
0.0242
0.0215
0.0199
0.0198
0.0228
Inspection of Figure 3 indicates that when survival is assigned a constant value
across treatments, the pattern of population growth differences across treatments
does not suggest an additional selenium-accentuated factor depressing population
growth rate. Population growth at 13 and 27 mg Se/kg is slightly higher than
might be expected from total egg production, when compared to lower
concentrations. The lack of influence of selenium exposure on timing of egg
production is also illustrated by comparing each treatment's cumulative
proportion of egg production over the course of the test, as shown in Figure 4.
Although the treatments differ somewhat in the temporal pattern of their egg
production, there is no consistent relationship with selenium exposure.
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1.2
c
o
a.
V)
-------
Chronic Value:
In other selenium studies, egg production and percent hatch have not generally
been thought to be related to selenium exposure. Although Besser et al. (2012)
noted that repeated measures ANOVA indicated a potential interaction between
selenium treatment and egg production on particular sampling dates, a thorough
examination of the study data from multiple perspectives indicates no statistically
significant or biologically apparent effect of selenium on egg production, timing
of egg production, or percent hatch at or below the highest tested concentration
of 27 mg Se/kg (dw). Likewise there was no discernible effect on deformity
rates.
In the separate tests of Fl larval survival at 21 days and of Fl juvenile-adult
survival at 58 days, the highest treatment, 27 mg Se/kg (dw), displayed lower
survival than any other treatment. Although the reduction was not sufficient to be
statistically significant, Besser et al. (2012) suggest that this is indicative of a
threshold. Note that among toxicity tests in general, the 10% effect level of the
ECio might or might not be statistically significant from the perspective of
hypothesis testing.
Shown below are the survival rates for the 27 mg Se/kg treatment adjusted to the
control (Abbott-adjusted), or similarly adjusted to the average survival at all
lower treatments (some of which had better survival than the controls). Either
way the adjustment is done, results are similar. (These survival data, Abbott-
adjusted, are included in Figure 2.)
27 mg Se/kg treatment:
adjusted to control
adjusted to all lower treatments
Larval
Surv at 21
days
92.9%
89.1%
Juv-Adlt
Surv at 58
days
87.5%
91.6%
The effect level at 27 mg Se/kg was thus 7% - 13% in the above comparisons.
While the concentration response curve is not sufficiently defined to allow
confident assignment of an ECio, the data suggest a chronic value in the general
neighborhood of 27 mg Se/kg.
An effect level of 27 mg Se/kg egg for the pupfish in this study is consistent with
the findings of Saiki et al. (2012a) who evaluated selenium in two related species
in the Salton Sea, California. These authors measured 3.09 to 30.4 mg/kg whole
body Se levels in mosquitofish and sailfin molliesand based on a lack of a
negative relationship with the catch-per-unit-effort deduced these species were
not adversely affected by selenium. They extrapolated the finding of selenium
tolerance to the pupfish based on the results of another study (Saiki et al 2012b)
in which mosquitofish and sailfin mollies accumulated similar levels of selenium
to the pupfish. Note: the ratio of selenium in whole body to egg tissues in the
pupfish was approximately 1:1 in the Besser study (see first table in the pupfish
study summary above).
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Staub, B.P. W.A. Hopkins, J. Novak, J.D. Congdon. 2004. Respiratory and reproductive characteristics
of eastern mosquitofish (Gambusia holbrooM) inhabiting a coal ash settling basin. Arch. Environ.
Contamin. Toxicol. 46:96-101.
Test Organism:
Exposure Route:
Study Design:
Effects Data:
Chronic Value:
Eastern mosquitofish (Gambusia holbrooM)
Waterborne and Dietary - field exposed
Fish were collected from a contaminated ash basin (ASH) and a reference pond
(REF)
In July 1999, male eastern mosquitofish were collected from ASH and REF
(n=26, n=20, respectively) for measurement of standard metabolic rate (SMR). In
July 1999, gravid female eastern mosquitofish were collected from ASH and
REF and transported to a laboratory for testing. To ensure all females were
fertilized in the field, all offspring used in testing were limited to three weeks
after collection. (Eastern mosquitofish are live-bearers with a four week gestation
period.) Response variables compared between ASH and REF were (1) SMR of
males, (2) brood size of females, (3) percent of live offspring at parturition, and
(4) trace element concentration in females and offspring.
SMRs of males, brood size of females, and offspring viability were not
significantly different between sites. Average (n=5) concentrations of selenium in
females were 11.85 and 0.61 mg/kg dw in ASH and REF sites respectively. The
average concentrations of selenium in offspring were 15.87 mg/kg dw and below
detection in ASH and REF sites, respectively. The authors point out that the
selenium concentrations are an under-estimate of the field levels since the
females were allowed to depurate during their time in the laboratory prior to
parturition.
>11.85 mg Se/kg dw whole body
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Saiki, M.K., B.A. Martin, and T.M. May. 2004. Reproductive status of western mosquitofish inhabiting
selenium-contaminated waters in the grassland water district, Merced County, California. Arch. Environ.
Contamin. Toxicol. 47:363-369.
Test Organism:
Exposure Route:
Study Design:
Effects Data:
Chronic Value:
Western mosquitofish (Gambusia affmis)
Waterborne and Dietary - field exposed
Fish were collected from selenium-contaminated sites and reference sites in the
San Joaquin River watershed.
Western mosquitofish were collected in June and July 2001 from San Luis Drain
(SLD) at Gun Club Road (Se-contaminated site), North Mud Slough at Gun Club
Road (MSN1; reference site); North Mud Slough at State Highway 140 (MSNs;
Se-contaminated site); San Joaquin River at Lander Avenue (SJR; reference site).
20 gravid females from each site were held in the laboratory for two weeks to
quantify live and dead births and to make other measurements. Only 17 females
from SLD were collected. Live and dead fry were visually examined under low
magnification with a binocular microscope for evidence of external abnormalities
(teratogenic symptoms such as spinal curvature, missing or deformed fins, eyes
and mouths and edema).
The percentage of live births was high at both Se-contaminated sites (96.6 to
99.9%) and reference sites (98.8 to 99.2%). There were no obvious anomalies
(e.g., deformities, edema) observed during the study. The concentration of
selenium in 4 postpartum females from the site with the highest selenium
concentration, SLD, ranged from 13.0 to 17.5 mg Se/kg dw (geometric mean of
the high and low is 15.1 mg Se/kg dw. The concentration of selenium of western
mosquitofish collected at each site is in Table D-8.
>15.1 mg Se/kg dw whole body
Table D-8. Selenium in whole body samples of western mosquitofish from study sites
Site
SLD
MSN2
MSN1
SJR
N
8
24
20
22
[Se], mg/kg dw
18.1
9.31
2.72
0.907
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Coughlan, D.J. and J.S. Velte. 1989. Dietary toxicity of selenium-contaminated red shiners to striped
bass. Trans. Am. Fish Soc. 118:400-408.
Test Organism:
Exposure Route:
Test Treatments:
Test Duration:
Study Design:
Effects Data:
Chronic Value:
Striped bass (Morone saxitilis; adults from Lake Norman, NC, approximately
250 g each)
dietary only
Treated fish were fed selenium contaminated red shiners (1 g) from Belews Lake,
NC (9.6 mg Se/kg ww or 38.6 mg Se/kg dw based on a mean reported moisture
content of 75.1 percent). Control fish were fed golden shiners from a local bait
dealer (0.3 mg Se/kg ww or 1.3 mg Se/kg dw based on a mean reported moisture
content of 76.3 percent).
Test treatments were as described above. Two tanks contained treated fish (n =
20 fish total), and one tank offish served as the control (n = 10 fish). Each tank
received a continuous flow of soft well water (hardness and alkalinity approx. 30
mg/L as CaCO3) throughout the exposure.
80 days
During the experiment, all striped bass (n = 10 per tank) were fed to satiation
three times per day. Pre-weighed rations of live red shiners (treated fish) and
golden shiners (controls) were added to the tanks and allowed 5 hours to feed.
Uneaten prey was removed and weighed. Composite whole-body samples of
each prey fish were collected at regular intervals throughout the study for whole-
body tissue selenium analysis. The final selenium concentration in epaxial white
muscle was determined for surviving striped bass at the end of the test. Moribund
striped bass were sacrificed so as to obtain muscle tissue samples for selenium
analysis. Samples of liver and trunk kidney of these and the surviving striped
bass were dissected for observations of histopathology.
Striped bass fed selenium-laden red shiners exhibited changes in behavior
(lethargy, reduced appetite), negligible weight gain, elevated selenium
concentrations in muscle, histological damage, and death. Control fish ate and
grew well, and behaved normally. Average selenium ingestion was between 60
and 140 Og Se/fish per day until day 30. Appetite of the treated fish appeared to
be significantly reduced beyond this point compared to the appetite of the control
group. By day 78, all striped bass fed the Se-laden red shiners either had died or
were moribund and sacrificed for analysis. The final selenium concentration in
muscle of treated striped bass averaged from 3.5 (tank 1) and 4.0 (tank 2) mg/kg
ww, or 16.2 and 18.5 mg/kg dw, respectively, assuming 78.4 percent moisture
content in muscle tissue; default May et al (2000) value for all species. The final
selenium concentration in muscle of control striped bass fed uncontaminated
golden shiners averaged 1.1 mg/kg ww, or 5.09 mg/kg dw (assuming 78.4
percent moisture content in muscle tissue; default May et al (2000) value for all
species).
The chronic value for percent survival of striped bass relative to final selenium in
muscle tissue after being fed Se-laden red shiners is <16.2 mg/kg dw.
An EC2o value could not be calculated for this data set because the data did not
meet the assumptions required for analysis.
Do not distribute, quote or cite
C-lll
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Bryson, W.T., W.R.Garrett, M.A. Mallin, K.A. MacPherson, W.E. Partin, and S.E. Woock. 1984.
Roxboro Steam Electric Plant 1982 Environmental Monitoring Studies, Volume II, Hyco Reservoir
Bioassay Studies. Environmental Technology Section. Carolina Power & Light Company.
28-day Embryo/Larval Study
Test Organism:
Exposure Route:
Study Design:
Effects Data:
Chronic Value:
Bluegill sunfish (Lepomis macrochirus; embryos and larvae)
dietary and waterborne - field exposure
Native adult bluegill were collected from Hyco Reservoir in Person County,
North Carolina and from a nearby control lake (Roxboro City Lake). Hyco
Reservoir is a cooling lake for Carolina Power & Light and receives the
discharge from the ash storage pond. No selenium values were given for Hyco
Reservoir, total selenium was not detected in the control lake (<1 (ig/L). A mean
selenium for the ash pond effluent from a previous study was 53 (ig/L (N=59;
range 35-80 (ig/L).
All combinations of crosses between the Hyco and control fish were made using
gametes from the collected fish. Fertilized eggs were exposed in egg cups to 0,
20 and 50 percent ash pond effluent under flow-through conditions. Percent hatch
and swim-up successes were measured. Swim-up larvae were released to
exposure tanks where there were fed zooplankton collected from Hyco and the
control lake. Larvae were observed for 28 days at which time survival and weight
were measured.
Survival to the swim-up stage was different between larvae from Hyco females
fertilized with either male type and those larvae from control females fertilized
with either male type. All crosses involving a Hyco female resulted in larvae
exhibiting 100 percent mortality prior to reaching swim-up. Percent survival
from hatch to 28 days for larvae from control females exposed to control water
and fed control lake zooplankton was only 5 and 12 percent for the two replicates
so no meaningful comparisons can be made to the different dilution exposures or
diet exposure. The mean concentrations of selenium in the ovaries, female liver
and female muscle were 49, 130, and 84 mg/kg dw, respectively.
Effect level: <49, <130 and <84 mg Se/kg dw in adult ovaries, liver and muscle,
respectively
<49.65 mg Se/kg dw in whole body using the muscle to whole body equation
<84 mg Se/kg dw maternal muscle
<49 mg Se/kg dw ovary
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Ingestion Study
Test Organism:
Exposure Route:
Study Design:
Effects Data:
Chronic Value:
Bluegill sunfish (Lepomis macrochirus; 30-day old larvae)
Dietary and waterborne - field exposed adults
Juvenile bluegill from crosses with females in 0, 20 and 50 percent ash pond
effluent were transferred to control water and fed zooplankton from either Hyco
or the control lake. Selenium in Hyco and control zooplankton was 45 and 1.9
mg/kg dw, respectively. Duration was not given.
Survival and observations on pathology and morphology were made in the two
diet treatments.
Mortality in larvae fed control zooplankton was 23.7 percent, whereas mortality
in larvae fed Hyco zooplankton was 97.3 percent. There were no differences in
survival (for two diet treatments) in larvae that were raised for the 30 days prior
to the test in different effluent concentrations (0, 20 50 percent). The average
selenium concentrations in the larvae fed control and Hyco zooplankton were 1.9
and 24.7 mg/kg dw, respectively.
Effect level for larval survival: <24.7 mg Se/kg dw in larvae
None recommended for larval tissue.
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Bryson, W.T., W.R.Garrett, M.A. Mallin, K.A. MacPherson, W.E. Partin, and S.E. Woock. 1985a.
Roxboro Steam Electric Plant Hyco Reservoir 1983 Bioassay Report. Environmental Services Section.
Carolina Power & Light Company. September 1985.
28-day Embryo/Larval Study
Test Organism:
Exposure Route:
Study Design:
Effects Data:
Bluegill sunfish (Lepomis macrochirus; embryos and larvae)
dietary and waterborne - field exposed
Resident adult bluegill were collected from Hyco Reservoir in Person County,
North Carolina and from a nearby control lake (Roxboro City Lake). Hyco
Reservoir is a cooling lake for Carolina Power & Light and receives the
discharge from the ash storage pond. For embryo/larval study up to swim-up
stage, control fish were collected from the unaffected portion of Hyco.
Repeat of 1982 28-day Embryo/Larval Study. Three crosses between: Hyco
female and Hyco male; control female with Hyco male; and control female with
control male. Gametes were fertilized and maintained for the 28-day test in ash
pond effluent dilutions of 0, 20 and 50 percent. Percent hatch, percent swim-up
success and survival were measured to 28 days post hatch. Two treatments were
replicated and fed zooplankton collected from Hyco-affected and Hyco-
unaffected (control). Larvae were observed for 28 days at which time survival
and weight were measured.
Embryo/Larval Study up to Swim-up Stage. Five crosses were made between fish
collected from the affected and unaffected areas. Percent hatch, percent swim-up
and survival were measured until swim-up (approximately 3-4 days after hatch).
28-day Embryo/Larval Study. All larvae that hatched from eggs obtained from
Hyco females died prior to completing swim-up (see table below).
Effect level (larval survival): <30, <33 and <59 mg Se/kg dw for adult female
bluegill in ovaries, liver and muscle, respectively
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Summary of 28-day embryo larval study
%
effluent
0
20
20
50
0
20
50
0
20
20
50
Parent
source in
cross
MXF
HXH
HXH
HXH
HXH
HXC
HXC
HXC
CXC
CXC
CXC
CXC
% hatch
92
98
92
97
89
96
60
79
90
88
72
% swim-
up
0
0
0
0
87
96
84
95
96
97
92
%
survival,
28-days
0
0
0
0
18
34
58
40
36
25
42
Adult tissue, mg Se/kg dw
Gonad
M
33
33
33
33
33
33
33
nd
nd
nd
nd
F
30
30
30
30
2.2
2.2
2.2
2.2
2.2
2.2
2.2
Liver
M
43
43
43
43
43
43
43
37
37
37
37
F
33
33
33
33
4.4
4.4
4.4
4.4
4.4
4.4
4.4
Muscle
M
62
62
62
62
62
62
62
27
27
27
27
F
59
59
59
59
2.7
2.7
2.7
2.7
2.7
2.7
2.7
Chronic Value:
<36.49 mg Se/kg dw in whole-body using the muscle to whole body equation.
<59 mg Se/kg dw muscle
<30 mg Se/kg dw ovary
Embryo/larval study to swim-up. Percent swim-up of larvae from parents
collected in non-affected Hyco averaged 93 percent, whereas percent swim-up
from larvae collected from affected Hyco was 12 percent. Effect levels were
determined for adult female and larval tissues. Larval tissues were averaged
across effluent concentrations (geometric mean).
Effect level (percent swim-up):
Adult female ovaries: >9.1 mg/kg dw; <30 mg/kg dw
Adult female liver: >26 mg/kg dw, <33 mg/kg dw
Adult female muscle: >25 mg/kg dw, <59 mg/kg dw
Larvae: >12.8 mg/kg dw; < 165 mg/kg dw
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Summary of Embyo/Larval Study up to Swim-up - Affected vs Unaffected Hyco
date
of
fert.
6-24
6-27
6-28
6-28
6-29
7-14
7-26
7-27
Parents'
capture
location in
Hyco
affected
affected
affected
affected
affected
unaffected
unaffected
unaffected
Percent hatch
at % effluent
0
93
99
29
98
88
92
99
76
20
98
88
34
86
93
80
94
84
50
94
77
35
91
85
84
93
86
Percent swim-up
at % effluent
0
0
0
25
5
59
79
100
100
20
0
0
14
0
42
92
98
89
50
0
0
3
0
25
89
98
91
Selenium in tissue, mg/kg dw
Adult female
Ovary
30
30
30
30
30
9.1
9.1
9.1
Liver
33
33
33
33
33
26
26
26
Muse
59
59
59
59
59
25
25
25
Larvae
0: 130
20: 120
0: 130
20: 120
0: 130
20: 120
0: 130
20: 120
0: 130
20: 120
0: 19
20: 11
50: 10
0: 19
20: 11
50: 10
0: 19
20: 11
50: 10
Chronic Value:
The chronic value estimated for the percentage larvae reaching the swim-up stage
is presented as a range:
>25 mg Se/kg dw (unaffected area) and <59 mg Se/kg dw muscle (affected area)
>30 mg Se/kg dw (unaffected area) and <9.1 mg Se/kg dw ovary (affected area)
Do not distribute, quote or cite
C-116
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Bryson, W.T., K.A. MacPherson, M.A. Mallin, W.E. Partin, and S.E. Woock. 1985b. Roxboro Steam
Electric Plant Hyco Reservoir 1984 Bioassay Report. Environmental Services Section. Carolina Power &
Light Company
Ingestion Study
Test Organism:
Exposure Route:
Test Treatments:
Test Duration:
Study Design:
Effects Data:
Chronic Value:
Bluegill sunfish (Lepomis macrochirus; juvenile- hatchery raised)
Dietary only
5 diets: Se form (nominal selenium concentration in base diet)
seleno-DL-cystine (5 mg/kg)
seleno-DL-cystine (10 mg/kg)
seleno-DL-methionine (5 mg/kg)
sodium selenite (5 mg/kg)
Hyco zooplankton (5 mg/kg)
60 days
Each treatment contained 40 fish which were maintained in a flow-through
system. Fish were fed at 3 percent of their body weight. Length and weight were
measured on days 30 and 60. Total selenium was measured in liver and whole-
body.
No decreased length or weight in any of the Se-diets relative to the control.
all values are whole-body
seleno-DL-cysteine: >2.16 mg Se/kg dw
seleno-DL-cysteine-2X: >3.74 mg Se/kg dw
seleno-DL-methionine: >2.46 mg Se/kg dw
sodium selenite : >1.21 mg Se/kg dw
Hyco zooplankton: >2.35 mg Se/kg dw
Because none of the selenium-spiked diet formulations affected growth of
juvenile fish at the concentrations tested, the chronic value selected for this study
is >3.74 mg Se/kg dw for the seleno-DL-cysteine-2X formulation.
Source and Exposure Embryo-Larval Study
Test Organism:
Exposure Route:
Test Treatments:
Test Duration:
Bluegill sunfish (Lepomis macrochirus; Adults from Hyco and a control lake)
Dietary and waterborne - field exposure
Four treatments:
Hyco-collected fish exposed to Hyco water in flow through spawning tanks.
Hyco-collected fish in control water in flow through spawning tanks.
Control fish exposed to Hyco water in flow through spawning tanks.
Hyco-collected fish in control water in flow through spawning tanks.
Adult fish were in spawning tanks 4-7 months
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Study Design:
Effects Data:
Eggs from each treatment were observed for percent hatch and percent swim-up.
Fish collected from the control lake did not spawn. Percent hatch and percent
swim-up from Hyco fish in Hyco and control water are given in the table below.
The percent hatch and percent swim-up were >83 and >83 for all the Hyco fish
suggesting no effect for these endpoints.
Source of
parents
Hyco
Hyco
Control
Control
Se in parental
liver tissue,
mg/kg dw
18.6
18.6
13.8
13.8
Water type
for eggs and
larvae
Hyco
well water
Hyco
well water
N
16
10
a
12
Percent hatch
86.6
83.8
a
86.0
Percent
swim-up
91.1
95.5
83.3
97.4
a percent hatch unknown.
Chronic Value: The chronic value for this study is > 18.6 mg Se/kg dw liver tissue.
Do not distribute, quote or cite
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Gillespie, R.B. and P.C. Baumann. 1986. Effects of high tissue concentrations of selenium on
reproduction by bluegills. Trans. Am. Fish. Soc. 115:208-213.
Test Organism:
Exposure Route:
Test Treatments:
Study Design:
Effects Data:
Chronic Value:
Bluegill sunfish, wild-caught (Lepomis macrochirus; adults; embryos and larvae)
dietary and waterborne - field exposure
High selenium adult fish were collected (electrofishing and with Fyke nets) from
Hyco Reservoir. Low selenium adult fish were collected from Roxboro City
Lake, Roxboro, NC.
All possible combinations of bluegill parents from Hyco Reservoir and Roxboro
City Lake were artificially crossed in June and July, 1982 and 1983, respectively.
Fertilization success was assessed by stripping subsamples of 100 to 500 eggs per
female and combining them with 2 ml of sperm. All zygotes were reared in
Roxboro City Lake water and percent fertilization was estimated 2-3 hours later
as the proportion of mitotically active zygotes. To estimate hatching success,
gametes were combined as before and subsamples of 100 to 300 embryos per
cross were transferred to egg cups and maintained in closed aquaria receiving re-
circulated Roxboro City Lake water. Percent hatch (approx. 2d at 22 to 25°C)
was based on the number of yolk-sac larvae. In 1982, about 200 embryos from 8
crosses were observed and preserved at intervals up to 40 h after fertilization, and
about 450 larvae were preserved at intervals of 40 to 180 h after fertilization. In
1983, about 1,800 larvae were observed and preserved from 40 to 150 hr from
crosses involving females from Hyco Reservoir, and about 40-300 hr for crosses
involving females from Roxboro City Lake (10 crosses total).
No significant differences were found in percent fertilization or in percent hatch
among parent combinations from the 18 crosses made in June 1982 and July
1983. In contrast, larvae from all crosses involving a Hyco female were
edematous; 100 percent of the larvae were abnormal in 7 of 8 crosses. Note: This
outcome was observed when the same female from Hyco Reservoir was crossed
with males from either Hyco Reservoir or Roxboro City Lake. The range of
selenium concentrations in the ovaries of Hyco Reservoir females used for the
cross experiments was from 5.79 to 8.00 (GM = 6.945 mg/kg ww; n=7). The
reported concentrations of selenium in ovaries and carcasses of females collected
from Hyco Reservoir in 1982 and 1983 were 6.96 and 5.91 mg/kg ww (n=22 and
28, respectively). The reported concentrations of selenium in ovaries and
carcasses of females collected from Roxboro City Lake in 1982 and 1983 were
0.66 and 0.37 mg/kg ww (n=14 and 19, respectively). The mean selenium
concentration in bluegill larvae (n=222) from artificial crosses of parents from
Hyco Reservoir was 28.20 mg Se/kg dw.
<46.30 mg Se/kg dw ovary using 85 percent moisture for ovaries measured in
study.
Do not distribute, quote or cite
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Doroshov, S., J. Van Eenennaam, C. Alexander, E. Hallen, H. Bailey, K. Kroll, and C. Restrepo.
1992. Development of Water Quality Criteria for Resident Aquatic Species of the San Joaquin River; Part
II, Bioaccumulation of Dietary Selenium and its Effects on Growth and Reproduction in Bluegill
(Lepomis macrochirus). Final Report to State Water Resources Control Board, State of California.
Contract Number 7-197-250-0.
Test Organism:
Exposure Route:
Test Duration:
Study Design:
Effects Data:
Bluegill sunfish (Lepomis macrochirus); Population A: selenium
bioaccumulation observations used 113 g (range 30-220 g) obtained from
Rainbow Ranch Fish Farm, California. Population B: spawning performance
observations used 106 g (range 65-220 g) females and 164 g (range 80-289 g)
males obtained from Chico Game Fish Farm.
Dietary only
Dietary
Seleno-L-methionine added to trout chow; the three nominal dietary
concentrations of 8, 18 and 28 mg/kg seleno-L-methionine were measured at 5.5,
13.9, and 21.4 mg/kg Se (moisture content 13 to 16%).
140 days
Population A fish and Population B females were fed nominal dietary treatments
8, 18 and 28 mg/kg seleno-L-methionine; Population B males were fed untreated
diets until the start of spawning. Population A fish were sampled on days 0, 30,
58, 86 and 114 for Se measurement. At least 3 females were sampled each event.
Fish remaining after day 114 were transferred to an outdoor pond fed untreated
diet and sampled on day 144 for depuration analysis.
On day 120 Population B males and females were paired for natural spawning
which had limited success. Fish were maintained in treatment tanks and females
were monitored for egg ripeness. When ripe, females were induced to ovulate
and ova were fertilized in vitro with semen stripped from males. Fertilized eggs
were sampled for fertilization success, Se content, and two live sub-samples for
bioassay, one a 30-day embryo-larval test and another for larval development
during first 5 days after hatching.
Larval development: after hatching, 100 larvae were transferred to beakers and
samples were examined daily for normal, abnormal and dead were recorded.
Larval bioassay: 90 fertilized eggs from each female were placed in groups of
approximately 30 eggs. Larvae and fry were fed rotifers and brine shrimp nauplii
through the 30 day observation.
Treatment effects were only observed on early development bioassays. In the 5-
day larval bioassay, systemic edema and underdeveloped lower jaw were
apparent in all larvae in the 21.4 mg/kg dietary treatment by day 3 and complete
mortality by day 5, except for two progenies where 10% of the larvae appeared
normal. No abnormalities were observed in control and 5.5 mg/kg treatment. 3 of
the 6 progenies in the 13.9 mg/kg treatment exhibited 10 to 20% larvae with
similar abnormalities (in table below). The average proportion of larvae with
edema were 5% in 13.9 and 95% in 21.9 mg/kg, both of these were statistically
different from the control (0% edema).
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Chronic Value:
In the 30-day larval survival bioassay, statistical difference was only in the
highest test treatment for survival and growth measurements, length and weight
(see table below). The proportion of abnormal larvae was higher in the selenium-
treated diets but was not significantly different from the control. The percent of
abnormal larvae in the 13.9 mg/kg treatment (7.2%) was only slightly higher than
the control (6.3%).
Authors present the effect level for bluegill at the 13.9 mg/kg dietary treatment
(NOEC 5.5 mg/kg) based on proportions of edema and delayed resorption of the
yolk sac. The latter endpoint is based on significantly greater yolk area and oil
globule area in the 13.9 and 21.4 mg/kg treated eggs.
The most sensitive endpoint, percent edema, as a function of selenium in egg dw
and adult muscle dw, was fitted to a logistic curve from which EC estimates were
calculated (see Figures 1 and 2). The ECio and EC2o values are given in the
following table.
Effect level
ECio
EC20
Egg, mg Se/kg dw
20.75
22.71
Maternal muscle, mg Se/kg dw
11.25
12.55
ECio value (edema) at 20.75 mg Se/kg egg dw or 11.25 mg Se/kg muscle dw
Chronic Value is 20.75 :g Se/g eggs dw.
Selenium Concentrations (mg/kg dw) in Bluegills from Population A Day 113 of Bioaccumulation
Dietary
treatment
Ovary
Female liver
Testis
Male liver
Control
2.17(0.05)
2.51(0.32)
2.65 (0.21)
4.10(0.37)
5.5 mg/kg dw
10.89(1.83)
NA
9.87
14.32
13.9 mg/kg dw
26.17(0.07)
22.75 (2.96)
16.38(0.71)
24.28 (4.54)
21.4 mg/kg dw
40.32 (2.44)
40.68(2.14)
29.70 (5.02)
52.47 (5.23)
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Selenium Concentrations (mg/kg dw) in Bluegill Parents (Population B) Used in Larval Toxicity
Tests
Dietary
treatment
Male liver
Testis
Female liver
Female muscle
Ovary
Eggs
Larvae
Fry
Control
4.07 (0.23)
1.87(0.11)
4.00 (0.26)
1.47 (0.14)
2.23(0.11)
2.81 (0.14)
NA
1.48(0.11)
5.5 mg/kg dw
6.94(1.58)
3.64 (0.47)
12.33 (1.09)
5.80 (0.79)
6.34 (0.47)
8.33 (0.63)
NA
1.25 (0.02)
13.9 mg/kg dw
20.46 (3.46)
9.96 (0.45)
25.98 (4.28)
10.41 (2.02)
14.10(2.62)
19.46 (3.83)
NA
1.37(0.06)
21.4 mg/kg dw
31.63(1.75)
15.25 (0.45)
47.60(4.11)
23.64 (2.04)
30.63 (3.23)
38.39 (3.14)
35.30(4.16)
1.46(0.03)
5-day Larval Development Toxicity Test (used for Chronic Value determination), average (SD)
Dietary
treatment
Free of Edema, %
Control
100
5.5 mg/kg dw
100
13.9 mg/kg dw
95 (2)*
21.4 mg/kg dw
4.3(2.7)*
Results from 30-day Embryo-larval Toxicity Test, average (SD)
Dietary treatment
Larval survival, %
Larval length, mm
Larval weight, mg
Abnormalities in
larvae, %
Control
71 (8.5)
19.1(1.2)
114(24)
6.3 (7.9)
5.5 mg/kg dw
51.9(26.5)
19.9(1.2)
133 (27)
15.0(5.8)
13.9 mg/kg dw
64.4 (3.4)
19.3 (0.8)
119(16)
7.2(3.1)
21.4 mg/kg dw
2.5 (3.5)*
16.6 (2.5)*
81 (37)*
25.0 (43.3)
* Statistically significantly different from control
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Thirty day toxicity test mortalities and tissue selenium concentrations in respective females, "n"
is number of eggs on Day 0, "r" is mortality on Day 30, "p" is proportions.
Progenies
08-2C
18-4C
5.5-1S
5.5-2S
5.5-6S
13.9-1S
13.9-3S
13.9-6S
21.4-1S
21.4-2S
21.4-3S
21.4-4S
21.4-5S
21.4-6S
21.4-7S
n
89
85
85
90
85
90
87
87
88
90
86
88
90
86
88
r
17
17
64
42
19
29
34
31
87
89
79
88
90
86
82
P
0.191
0.200
0.753
0.467
0.224
0.322
0.391
0.356
0.989
0.989
0.919
1.000
1.000
1.000
0.932
[Se], mg/kg dw (female)
Ovary
1.95
2.38
7.72
5.55
4.06
3.94
21.82
20.40
29.90
45.82
27.24
23.18
32.64
37.63
18.02
Liver
4.04
5.03
14.89
7.06
10.49
7.54
34.74
36.82
38.02
33.96
59.01
62.71
55.25
48.14
36.10
Muscle
2.25
0.95
7.07
5.80
1.41
2.75
15.44
16.58
NA
31.10
17.28
27.40
24.00
24.66
17.42
Eggs
3.54
3.25
11.49
8.31
6.18
8.55
22.06
30.20
44.02
36.31
25.21
52.18
42.40
38.47
30.12
Do not distribute, quote or cite
C-123
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CO
Q
o
CO
CD
0.
90
80
70
60
"co 50
"co
| 40
co
_ 30
20
10
.4 .6 .8 1.0 1.2
Log(mg Se/kg egg dw)
Thirty-day survival of offspring of respective females shown in previous table, along with one
possible non-convergent solution. TRAP version 1.22 could not converge to a single solution, and a
range of solutions are possible, none of which can reconcile the disparate observations. This
particular solution yielded a survival ECio of 22.75 mg Se/kg egg dw, only slightly above the edema
(measured at 5 days) shown below.
Do not distribute, quote or cite
C-124
Draft Document
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MED Toxic Response Analysis Model 01/07/2015 18:10
E
•o
120
100
60
CL
20
.8 1.0 1.2
Log{mg Se/kg egg dw)
1.8
Parameter Summary (Threshold Sigmoid Regression Analysis)
Parameter Guess FinalEst StdError 95%LCL 95%UCL
LogX50 1.442S 1.4342 0.0000 1.4342 1.4342
S 5.452 4.712 O.DOO 4.712 4.712
YO 98.33 100.00 0.00 100.00 100.00
% Effect
50.0
20.0
10.0
5.0
Effect Concentration Summary
Xp Est 95%LCL
27.1S
22.71
2075
19.460
85%UCL
MED T.jsidly ^dainshc Ariljsis Voasl. Version 1 ;t
Do not distribute, quote or cite
C-125
Draft Document
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MED Toxic Response Analysis Model 01/07/2015 18:12
E
•o
120
100
60
CL
20
.6 .6 1.0 1.2
Log{mg Se/kg muscle dw)
1.4
1.6
Parameter Summary (Threshold Sigmoid Regression Analysis)
Parameter Guess FinalEst StdError 95%LCL 95%UCL
LogX50 1.2029 1.1926 0.0000 1.1926 1.1926
S 4.516 3.904 O.DOO 3.904 3.904
YO 98.33 100.00 0.00 100.00 100.00
% Effect
50.0
20.0
10.0
5.0
Effect Concentration Summary
Xp Est 95%LCL
15.581
12.545
11.246
10.410
95%UCL
c Ariljsis VoasL Version 1 ;t
Do not distribute, quote or cite
C-126
Draft Document
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Hermanutz et al. 1992. Effects of elevated selenium concentrations on bluegills (Lepomis macrochirus)
in outdoor experimental streams. Environ. Tox. & Chem. 11: 217-224
Hermanutz et al. 1996. Exposure of bluegill (Lepomis macrochirus) to selenium in outdoor experimental
streams. U.S. EPA Report. Mid-Continent Ecology Division. Duluth, MN.
Tao, J., P. Kellar and W. Warren-Hicks. 1999. Statistical Analysis of Selenium Toxicity Data. Report
submitted for U.S. EPA, Health and Ecological Criteria Div. The Cadmus Group.
Test Organism:
Exposure Route:
Study Design:
Bluegill (Lepomis macrochirus; 3 to 4-year old adults)
Dietary and waterborne followed by dietary only
Dietary and waterborne
Selenite was added to artificial streams which entered the food web; thus, fish
were also exposed to selenium in the diet.
Dietary only
Recovering streams exposed bluegill to selenium in prey organisms. Selenite
addition to water was ceased (selenium in water was below detection level).
Eight Monticello artificial streams were used for three separate studies between
1987 and 1990.
Table 1. Study Design.
Stream
Dates
BGa put in station 0-2
BG transferred to sta.
6 End of study
1
2
3
4
5
6
7
8
Study I
9-1-87
5-16-88
8-22-88
Unused
Unused
10 ng/L
30 ng/L
Control
30 ng/L
Control
10 ng/L
Study II
10-88
5-89
8-89
Control
2.5 ng/L
10 ng/L
Recovering
Control
Recovering
2.5 ng/L
10 ng/L
Study III
11-89
5-90
7-90
Control
Recovering
Recovering
Recovering
Control
Recovering
Recovering
Recovering
BG = Bluegill
Do not distribute, quote or cite
C-127
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The design of the three Hermanutz et al. studies is included in Table 1 and a schematic diagram of an
artificial stream is provided below (Figure 1). For each study, a random sample of 22-50 adult bluegill
were transferred from stations 0-2 (provided temperatures above 4°C during winter) to station 6 (most
suitable for nests) during mid-May for spawning. Spawning activity was monitored in the streams.
Embryo and larval observations were made in situ and in the laboratory from fertilized eggs taken from
the streams and incubated in the lab.
Figure 1. Schematic Design of One of the Artificial Streams in the Monticello Study
Station Number
inlet
I
Adults from fall to
mid-May
Adult barrier-
Adult barrier
Adult barrier •
Adults from mid-
, May to end of study
Do not distribute, quote or cite
C-128
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Table 2. Effects on Progeny - Study Ia
Egg cup observations
treatment
control
control
10 ng/L
10 ng/L
30 ng/L
stream
5
7
3
8
4
ovary Se (mg/kg ww)
Up
NA
0.47
4.29
4.72
3.71
down
0.53
0.01
2.53
6.37
NA
geomean
0.53
0.07
3.29
5.48
3.71
ovary Se
(mg/kg
dw)b
2.21
0.29
13.73
22.85
15.46
Geomean
ovary Se
(mg/kg
dw)
0.79
17.71
15.46
% hatch
mean ± SD
93.3 ±9.1
71.5 ±22.5
60.3 ±25. 8
% survival
to 4th day
mean ± SD
69.7 ±13.9
28.8 ±23.1
9.1 ± 12.9
% edema
mean ± SD
0.1 ±0.2
80 ±1.0
50.3 ±64.1
% lordosis
mean ± SD
1.8 ±2.6
11.6±15.9
6.3 ± 1.8
% hemorr
mean ± SD
0.1 ±0.3
28.5 ±40.6
26.8 ±20.2
Nest observations
treatment
control
control
10 ng/L
10 ng/L
30 ng/L c
stream
5
7
3
8
4
ovary Se (mg/kg ww)
up
NA
0.47
4.29
4.72
3.71
down
0.53
0.01
2.53
6.37
NA
geomean
0.53
0.07
3.29
5.48
3.71
ovary Se
(mg/kg
dw)b
2.21
0.29
13.73
22.85
15.46
Geomean
ovary Se
(mg/kg
dw)
0.79
17.71
15.46
# active
nests
mean ± SD
6.5 ±2.1
5.0 ±4.2
1.0± 1.4
# embryos
Collected
mean ± SD
1441 ±205
1282 ±457
361±510
% dead
Embryos
mean ± SD
0.9 ±0.03
3.2 ±2.9
0.4
# larvae
Collected
mean ± SD
3947 ±1888
1169 ±1093
157 ±222
% dead
Larvae
mean ± SD
3.0±1.1
17.0±21.3
12.1
a Selenium concentrations in table were taken from Hermanutz et al. (1996); effect values were taken from Hermanutz et al (1992).
b used 76% moisture for egg/ovary in bluegill (average of Gillespie and Bauman 1986 and Nakamoto and Hassler 1992) to convert egg/ovary ww
to dw
0 No active nests, embryos, or larvae found in one of the 30 (ig/L streams. Therefore, N = 1 for % dead embryos and dead larvae in the 30 ng/L
treatment
Do not distribute, quote or cite
C-129
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Table 3. Effects on Progeny - Study IIa
Egg cup observations
treatment
control
control
2.5 ng/L
2.5 ng/L
10 ng/L
10 ng/L
rec 30 ng/L
rec 30 ng/L
stream
1
5
2
7
3
8
4
6
No. of
trials
6
5
0
4
3
2
0
6
%
hatch
93.0
96.4
NA
81.4
83.3
91.1
NA
92.9
%
survival
to 3rd
day
75.2
71.5
NA
71.6
57.7
57.1
NA
73.0
%
edema
0
0
NA
0
100
100
NA
17.4
%
lordosis
0
0
NA
0
11.1
18.2
NA
0
% hem or r
0
0
NA
3.6
49.3
41.1
NA
11.5
% healthy"
97.8
97.9
NA
92.2
0
0
NA
70.7
ovary Se (mg/kg ww)
up
1.02
1.09
2.02
6.96
5.87
down
0.78
0.76
1.82
3.36
8.1
12.6
13.2
avg
0.89
0.91
1.82
2.61
8.10
9.36
8.80
ovary Se
(mg/kg dw)c
3.72
3.79
7.58
10.86
33.75
39.02
36.68
Nest Observations
Treatment
control
control
2.5 ng/L
2.5 ng/L
10 ng/L
10 ng/L
R30ng/L
R30ng/L
Stream
1
5
2
7
3
8
4
6
#
active
Nests
6
9
1
5
2
3
0
8
#
embryos
Collected
2458
1329
0
1462
672
931
NA
646
% dead
embryos
0.94
0
0
0
0.32
NA
0
# larvae
collected
3252
3435
2497
4717
5376
750
NA
6782
%
dead
larvae
0.03
1.05
0.20
0.08
0.50
0.40
NA
7.8
#samples
w larvae
7
13
3
8
9
4
NA
16
%
edema
0
0
4.1
0
81.4
50
NA
27.3
%
lordosis
0
0
25
0
5.0
14.7
NA
0
%
hemorr
0
0
77.6
52
55.5
26.7
NA
17.1
ovary Se (mg/kg ww)
up
1.02
1.09
2.02
6.96
5.87
Down
0.78
0.76
1.82
3.36
8.1
12.6
13.2
Avg
0.89
0.91
1.82
2.61
8.10
9.36
8.80
ovary Se
(mg/kg
dw)c
3.72
3.79
7.58
10.86
33.75
39.02
36.68
a Selenium concentrations in table were taken from Hermanutz et al. (1996); effect values were taken from Tao et al. (1999).
b Among live larvae that survived up to third day after first larvae hatched; assumes the observations of multiple abnormality types always co-
occurred in the same organism. This may overestimate the actual % healthy when this assumption is violated.
0 used 76% moisture for egg/ovary in bluegill (average of Gillespie and Bauman 1986 and Nakamoto and Hassler 1992) to convert egg/ovary ww
to dw
R = recovering stream
Do not distribute, quote or cite
C-130
Draft Document
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Table 4. Effects on Progeny - Study III3
Egg cup observations
treatment
control
control
R2.5 ng/L
R2.5 ng/L
R 10 ng/L
R 10 ng/L
R 30 ng/L
R 30 ng/L
Stream
1
5
2
7
3
8
4
6
number of
trials
2
3
3
6
3
5
% hatch
92
76.7
87.3
87.2
75.3
92
% survival
to 3rd day
58.6
69.2
66
76.5
74.5
78
% edema
0
0
0
0
0
% lordosis
0
0.9
0
0
0
% hemorr
0
0.8
0
0
0
ovary Se
(mg/kg ww)
1.2
0.93
1.84
1.97
6.25
2.44
3.82
ovary Se (mg/kg
dw)b
5.0
3.88
7.67
8.21
26.04
10.17
15.92
Nest observations
treatment
control
control
R2.5 ng/L
R2.5 ng/L
R 10 ng/L
R 10 ng/L
R 30 ng/L
R 30 ng/L
stream
1
5
2
7
3
8
4
6
# active
nests
2
2
5
5
2
4
9
# samples
with larvae
5
3
5
2
4
4
13
% edema
0
0
0
0
0
0
0
% lordosis
0
0
0
0
0
0
0
% hemorr
0
0
0
0
0
0
0
ovary Se
(mg/kg ww)
1.2
0.93
1.84
1.97
6.25
2.44
3.82
ovary Se
(mg/kg dw)b
5.0
3.88
7.67
8.21
26.04
10.17
15.92
a The NOAEC for the study are from recovering 30 Og Se/L treatment.
b used 76% moisture for egg/ovary in bluegill (average of Gillespie and Bauman 1986 and Nakamoto and Hassler 1992) to convert egg/ovary ww
to dw
R = recovering stream
Do not distribute, quote or cite
C-131
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Effects Data: Tables 2 through 4 include exposure and effects data for Study I,
II, and III, respectively. Study I&II deformity and survival data reported in the
tables above from the nest and egg in response to Se concentrations in parental
ovaries (mg/kg dw) were compiled in Table 5 for TRAP analysis. Study I effects
data were obtained from Hermanutz et al. (1992), and corresponding Study I
ovary Se concentrations were obtained from Hermanutz et al. (1996). Study II
effects and exposure data were obtained from Hermanutz et al. (1996). Neither
data from the two recovering streams in Study II, nor any of the Study III data
were included in these analyses. As stated in the main text of the criteria
document, the recovery streams do not reflect the type of system to which water
quality criteria are most commonly applied; those receiving existing waterborne
pollutant discharges.
Prior to analysis, all percentages were transformed (100-% value of response) so
that the response variables decreased with increasing Se. In the initial analysis,
the EC 10 for % larvae free from edema in response to Se concentrations in
ovaries was analyzed in TRAP by combining nest and egg cup observations in
Study II with egg cup data from study I (Figure 2). The resulting EC 10, which
was presented in the 2014 draft criterion document, was 12.68 mg/kg dw. As
shown in Figure 2, the model fit is poor, with the declining limb passing through
two values from egg cup data at intermediate Se concentrations and ignoring two
values from nest data at relatively high Se concentrations.
Next, data were reanalyzed to determine whether a different endpoint would
result in a more appropriate concentration-response model. A comparison of the
% larval survival, % edema and % normal survival endpoints are illustrated in
Figures 3 and 4. The relationship between each of these endpoints and selenium
in water shows a clear relationship with minimal variation (Figure 3). When
these endpoints are plotted against selenium in ovaries, a similar relationship
exists but with more variability (Figure 4). The reason for the increased variation
is due to inconsistent bioaccumulation between S1 and S2 studies (see Figure 4
bottom). This inconsistency is illustrated in Figure 5 which shows the bluegills
in S2 accumulated more selenium in their ovaries compared to the bluegill in S1.
The effects vs Se in ovary plots generally show effects occurring between 10 and
16 mg/kg Se.
The bottom panel of Figure 4 shows the same data as Figure 2, and shows that
the nest data add to the variability of the edema data. When the survival and
edema data are combined into a single endpoint, only the egg cup data can be
used, because survival was only evaluated for the egg cup data. Using the
combined endpoint (Figure 4, middle panel) removes the variability introduced
by the nest data.
Figure 6 shows the relationship between the combined survival+deformity
endpoint in response to selenium in ovaries. The combined endpoint was defined
as the product of larval survival and the lesser of "1-edema" or "1-lordosis" for
all egg cup data from Study I and II (Table 5). The incidence of edema was
greater than lordosis for all but 1 treatment (Study I, stream 5,7), where edema
0.1% and lordosis was 1.8%. Percent normal survival was the most sensitive
endpoint, with an EC 10 of 11.36 mg/kg dw.
Do not distribute, quote or cite C-132 Draft Document
-------
In addition to the combined endpoint, EC 10s for individual endpoints were also
calculated in TRAP. The results for % free from edema were described
previously (Figure 2). The EC10 for % lordosis was 19.38 mg/kg ovary Se dw,
but because of the large standard error surrounding the slope and inflection point
parameters of the model, the corresponding confidence intervals (0.06-6103
mg/kg ovary Se dw) were extremely large. Coupled with the fact that the
incidence of lordosis was low in both studies (less than 20% at the highest Se
concentrations, with a maximum value of 25%), this endpoint was determined to
be less appropriate than %edema. Model convergence for % hemorrhage could
only be achieved in TRAP at an unrealistic y-intercept value of 150, indicating a
negative 50% incidence of hemorrhage at a Se concentration of zero. Finally,
model convergence for %larval survival could not be achieved in TRAP.
Chronic Value: The chronic value for bluegill was calculated as the ECi0 value of 11.36 mg/kg
Se dw (larval survival + free from edema combined endpoint in response to Se
concentration in the parental ovaries).
Do not distribute, quote or cite C-133 Draft Document
-------
Table 5. Combined Nest and Egg Cup Observations for TRAP Analysis (Study I&II)
Study
II
II
II
II
II
II
II
II
II
II
II
II
I
I
I
Stream
1
1
5
5
2
7
7
7
3
3
8
8
5,7
3,8
4
Data
Source
Egg Cup
Nest
Egg Cup
Nest
Egg Cup
Nest
Egg Cup
Nest
Egg Cup
Nest
Egg Cup
Nest
Egg Cup
Egg Cup
Egg Cup
Se Treatment
Control
Control
Control
Control
2.5 Mg/L
2.5 ng/L
2.5 Mg/L
2.5 ng/L
10 ng/L
10 Mg/L
10 Mg/L
10 Mg/L
Control
10 Mg/L
30 Mg/L
Ovary Se
(mg/kg dw)
3.72
3.72
3.79
3.79
7.58
7.58
10.86
10.86
33.75
33.75
39.02
39.02
0.79
17.71
15.46
%Edema
0
0
0
0
NA
4.1
0
0
100
81.4
100
50
0.1
80
50.3
%Lordosis
0
0
0
0
NA
25
0
0
11.1
5.0
18.2
14.7
1.8
11.6
6.3
% Hemorrhage
0
0
0
0
NA
77.6
3.6
52
49.3
55.5
41.1
26.7
0.1
28.5
26.8
%Survival
75.2
71.5
71.6
57.7
57.1
69.7
28.8
9.1
Do not distribute, quote or cite
C-134
Draft Document
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co
120 r
100f
80
co
E
CD
73
60
40
<•- 20
.2 .4 .6 .8 1.0 1.2
Log Ovary Se(mg/kg dw)
1.4
1.6
1.8
Model Parameters (Logistic Regression Nonlinear Regression Model)
Initial Guess Final Estimate S.E. 95% LCL 95% UCL
logX50
S
Y-intercept
1.2
6.29
100
1.1905
6.2904
99.93
2.52E-02
4.6679
6.0437
1.135
-3.9836
86.628
1.2459
16.564
113.23
Effect Concentration Summary
% Effect ECx 95% LCL 95% UCL
50
20
10
5
15.504
13.657
12.68
11.842
13.646
10.232
8.4743
7.1031
17.615
18.229
18.974
19.743
Figure 2. Incidence of larval bluegill edema as a function of the logarithm of the selenium
concentration in parental ovaries. Study I and II egg cup and nest data combined. The data points are
the same as the lower panel of Figure 4.
Do not distribute, quote or cite
C-135
Draft Document
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80
70
50
40
SI cup
20 [[[ BS2cup
10 [[[
o [[[
1 2 4 8 16 32
Water Cone. |jg/L
80
« 70
1 60
•£ SI cup
| 20 "S2CUP
^ 10
o • •
-------
80
70
- 60
I 50
40
on SI CUp
20 BS2 cup
10
o .........
0.5 1 2 4 8 16 32 64
mg Se/kg Ovary dw
80
s 70
o
o 60
•v 50
M40
I 30 SI cup
E 20 HS2 cup
0.5 1 2 4 8 16 32 64
mg Se/kg Ovary dw
120
100 - • M • n
SB
| 80
S 60 SI cup
•
§ 40 • BS2 cup
20 S2 nest
0 - : • : • •. • •.
0.5 1 2 4 8 16 32 64
mg Se/kg Ovary dw
Figure 4. Percent 3-day larval survival vs Se in ovary (top); % surviving and normal (combined
larval survival and larvae without edema) vs Se in ovary (middle); 100-%edema vs Se in ovary
(bottom). SI = study I; S2 = study II (see page 1 above for description).
Do not distribute, quote or cite C-137 Draft Document
-------
M
S 20
b
« 10
SI
IS2
4 8
Water |jg/L
16
32
Figure 5. Se in ovary versus Se in water.
Do not distribute, quote or cite
C-138
Draft Document
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90
80
Q
"O
c
CO
"to
_>
>
^5
CO
"O
-------
Coyle, J.J., D.R. Buckler and C.G. Ingersoll. 1993. Effect of dietary selenium on the reproductive
success of bluegills (Lepomis macrochirus). Environ. Toxicol. Chem. 12:551-565.
Test Organism:
Exposure Route:
Test Duration:
Study Design:
Bluegill sunfish {Lepomis macrochirus; two-year old pond-reared adult fish and
resultant fry)
Dietary and waterborne
Dietary
Seleno-L-methionine added in an aqueous solution to Oregon moist pellets;
moisture content of diet was 25 percent.
Waterborne
Flow through, 10 (ig Se/L nominal, 6:1 ratio of selenate:selenite, 98 percent
purity, adjusted to pH 2 with HC1 to prevent bacterial growth and change in
oxidation states of Se(IV) and Se(VI).
140 days
The experiment consisted of a test control and food control (see Test Treatment
table below) with fish (n=28 initially) in the four remaining treatments fed one of
the four seleno-methionine diets in combination with 10 (ig Se/L in water.
Spawning frequency, fecundity, and percentage hatch were monitored during the
last 80 days of the exposure period. Survival of resulting fry (n=20) was
monitored for 30 days after hatch. Adults and fry were exposed in separate,
modified proportional flow-through diluters. Fry were exposed to the same
waterborne selenium concentrations as their parents. Adults were fed twice daily
ad libitum. Whole-body selenium concentrations in adult fish were measured at
days 0, 60, and were calculated from individually analyzed carcass and gonadal
tissue (ovaries and testes) at day 140. Eggs not used in percentage of hatch
determinations were frozen and analyzed for total selenium.
Measured Se in:
water
(Hg Se/L)
diet
(mg Se/kg dw)
Test Treatments
1
(test control)
0.56
0.76
2
(food control)
8.4
0.76
3
10.5
4.63
4
10.5
8.45
5
10.1
16.8
6
11.0
33.3
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C-140
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Effects Data:
There was no effect of the combination of highest dietary selenium concentration
(33.3 mg/kg dw) in conjunction with exposure to a waterborne selenium
concentration of 11.0 (ig/L on adult growth (length and weight), condition factor,
gonad weight, gonadal somatic index, or reproductive endpoints (i.e., spawning
frequency, number of eggs per spawn, percentage hatch) during the 140-day
exposure. The mean corresponding whole-body selenium concentration in adults
exposed to this waterborne and dietary selenium combination was 19 mg/kg dw.
Survival of fry from the exposed adults was affected by 5 days post-hatch.
Concentrations of whole-body selenium in adult tissue at day 60 were used to
determine effects in the fry because eggs were taken for the larval tests beginning
at day 60 of the adult exposure.
Effects on Adults
Se in diet,
mg/kg dw
0.8
0.8
4.6
8.4
16.8
33.3
Se in water,
Mg/L
0.5
7.9
10.5
10.5
10.1
10.1
whole -body
Se (140 d),
mg/kg dw
0.8
1.0
3.4
6.0
10
19
replicate
A
B
A
B
A
B
A
B
A
B
A
B
total no.
spawns
15
10
12
11
20
12
2
9
13
13
14
4
eggs/spawn
14,099
5,961
9,267
9,255
9,782
13,032
10,614
7,995
10,797
9,147
8,850
8,850
hatchability,
%
94.5
90.5
89.5
84.5
86.5
96.5
96.5
90
83
91.5
80
80
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Effects on Larvae
Se in diet, mg/kg
dw
0.8
0.8
4.6
8.4
16.8
33.3
Se in water, (ig/L
0.5
7.9
10.5
10.5
10.1
10.1
egg, mg/kg dw
1.8
1.8
7.3
13
23
42
adult whole -body
(60 d), mg/kg dw
0.9
0.9
2.9
4.9
7.2
16
mean survival,
%
92
93
90
95
87
7
Chronic Value:
EC20 and ECi0 estimates using logistic equation with log transformation of
exposure:
effect level
EC20
ECio
egg, mg Se/kg dw
26.30
24.10
whole body, mg Se/kg dw
8.954
7.936
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C-142
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Coyle et al. 1993 bluegill larval survival - TRAP logistic
110
100
90
80
_- 70
•5 60
m 50
CO
1 40
30
20
10
0
c
r
£ f
,2 .4 .6 .8 1.0 1.2 1.4 1.6 1.8
Log([Se]egg mg/kg dw)
Parameter Summary (Logistic Equation Regression Analysis)
Parameter Guess FinalEst StdError 95%LCL 95%UCL
LogXSO 1.4626 1.4990 0.0137 1.4554 1.5427
S 1.8812 5.041 0.529 3.358 6.723
YO 92.50 92.50 1.05 89.15 95.85
Effect Concentration Summary
% Effect
50.0
20.0
10.0
5.0
Xp Est
31.55
26.93
24.55
22.54
95%LCL
28.54
23.75
21.19
19.02
95%UCL
34.89
30.54
28.45
26.72
06/29/2009 14:04
MED Toxic Response Analysis Model, Version 1.03
Do not distribute, quote or cite
C-143
Draft Document
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Coyle et al. 1993 bluegill larval survival - TRAP logistic
110
100
gd1
80
- 70
03
•| 60
" 50
CO
1 «
30
20
10
0
c
r-
m
• ~^^>_
: \
; \
\
\
V^_
.2 .4 .6 .8 1.0 1.2 1.4
Log([Se]adult whole body mg/kg dw)
Parameter Summary (Logistic Equation Regression Analysis)
Parameter Guess FinalEst StdError 95%LCL 95%UCL
LogXSO 1.0102 1.0408 0.0202 0.9764 1.1051
S 1.9483 3.851 0.452 2.412 5.290
YO 92.50 92.52 1.17 88.81 96.23
Effect Concentration Summary
% Effect
50.0
20.0
10.0
5.0
Xp Est
10.985
8.929
7.910
7.074
95%LCL
9.472
7.398
6.342
5.484
95%UCL
12.739
10.777
9.865
9.125
06/29/2009 14:10
MED Toxic Response Analysis Mode!, Version t.03
Do not distribute, quote or cite
C-144
Draft Document
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Cleveland, L. et al. 1993. Toxicity and bioaccumulation of waterborne and dietary selenium in juvenile
bluegill sunfish (Lepomis macrochirus}. Aquatic Toxicol. 27:265-280.
Test Organism:
Life Stage:
Exposure Route:
Study Design:
Effects Data:
Bluegill sunfish {Lepomis macrochirus)
juvenile (5 months - waterborne exposure; 3 months - dietary exposure)
waterborne (60-d) and dietary (90-d) - separate exposures
waterborne - 6:1 selenate:selenite at 0.17, 0.34, 0.68, 1.38, 2.73 mg/L; dietary -
seleno-L-methionine in Oregon moist at 1.63, 3.25, 6.5, 13, 26 mg Se/kg dw)
Fish were exposed using a flow-through diluter. Each test consisted of an
exposure and a depuration phase. Whole body tissue measurements were made at
31 and 60 days of waterborne exposure and at 31, 59 and 90 days of dietary
exposure. Mortality and condition factor, K (weight x 105/length3), were reported
at selected intervals.
The waterborne exposure (see table below) was determined to have an EC2o =
4.07 mg Se/kg dw (1.96-8.44 mg/kg 95% CL). However, because it was a water-
only exposure, it was not considered in the derivation of the FCV. These data
nevertheless provide evidence that exposure route influences the tissue
concentration toxicity threshold, although the mechanistic explanation for this
phenomenon is lacking.
A mortality effect level for the dietary exposure could not be calculated because
the highest selenium whole body concentration (13.4 mg Se/kg dw) only had
17.5% mortality. The middle selenium concentration did have 22.5% mortality.
Cleveland et al. reported a significant decrease in K between 4.7 and 7.7 mg/kg
dw (see table below).
Waterborne Exposure Study
Measured selenium in
water (:g/L)
20 (control)
160
330
640
1120
2800
60-d measured
selenium in whole
body (mg/kg dw)
1.1
2.8
4
5.3
9.8
14.7*
60-d mortality (%)
10
12.5
22.5
52.5
70
97.5
Condition factor (K)
1.5
1.5
1.6
1.5
1.6
NA
*a 30-d measurement because all fish were dead at 60 days in this concentration.
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C-145
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Dietary Exposure Study
Measured selenium in
food (mg/kg ww)
0.68 (control)
2.3
3.5
6.6
12.7
25
90-d measured
selenium in whole
body (mg/kg dw)
1
2.1
3.3
4.7
7.7
13.4
90-d mortality (%)
5
7.5
10
22.5
15
17.5
Condition factor (K)
1.3
1.3
1.3
1.3
1.2
1.2
Discussion
The study demonstrates the influence of exposure route on the potency of a given
tissue concentration, as shown in the figure. The TRAP threshold sigmoid
concentration-response curve for the water-only exposure yields an EC50 of 6.5
mg Se/kg dw WB. In contrast, higher whole-body concentrations acquired via
diet did not yield significant effects and cannot support a TRAP-fitted
concentration-response curve or EC estimate. Examination of the graph indicates
that the water-only concentration-response curve would need to be shifted to the
right a minimum of 4-fold (or possibly more) to be able to fit the (lack of) effects
observed in the dietary study. This supports the decision to derive the criteria
only from studies relying on the environmentally relevant exposure route, diet.
sp
OS
•n
$
£
3
1/1
100 -,
90 "
80
70 4
60 4
50 H
40 -|
30 J
20 -j
10 -|
01
t
1
• n
B * •
Via water exposure, 60-d
• Via dietary exposure, 90-d
Model for water exposure
2 4 8 16 32
Whole Body Concentration, mg Se/kg dw
Survival at 60-days (for water exposure) or 90-days (for dietary exposure) versus
whole-body concentration.
Do not distribute, quote or cite
C-146
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Chronic Value: Given (a) the very slight reduction in K (1.3 to 1.2 between 4.7 and 7.7 mg Se/kg
dw WB, with no further reduction at 13.4 mg Se/kg dw WB) and uncertain
relevance of growth data, and (b) no apparent concentration-related effect on
mortality between 4.7 and 13.4 mg Se/kg dw WB, the NOAEC is interpreted to
be 13.4 mg Se/kg dw for this study; and the chronic value is >13.4 mg Se/kg dw
whole body.
Do not distribute, quote or cite C-147 Draft Document
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Lemly, A.D. 1993a. Metabolic stress during winter increases the toxicity of selenium to fish. Aquatic
Toxicol. 27:133-158.
Test Organism:
Exposure Route:
Test Duration:
Study Design:
Effects Data :
Chronic Value:
Comments:
Bluegill sunfish (Lepomis macrochirus; juvenile 50-70 mm)
Waterborne and dietary
Water
1:1 selenite:selenate in stock at pH 2; metered in to reach 5 :g/L
Diet
seleno-L-methionine in TetraMin (5 mg/kg dw)
180 days
Fish were exposed (treatment and control) under intermittent flow-through
conditions for 180 days. Tests were run at 4° and 20°C with biological
(histological, hematological, metabolic and survival) and selenium measurements
made at 0, 60, 120 and 180 days. Fish were fed at a rate of 3% body weight per
day. All treatments were initiated at 20°C and then decreased in the cold
treatment at a rate of 2°C per week for 8 weeks to reach 4°C and then maintained
at that temperature for the remainder of the 180 days.
In the 20°C test, fish accumulated 6 mg/kg dw selenium (whole-body) with no
significant effect on survival (4.3% and 7.4% mortality in control and treatment,
respectively). In the 4°C test, fish exposed to selenium accumulated 7.9 mg/kg
dw (whole-body) selenium and had significant mortality after 120 (33.6%) and
180 days (40.4%) relative to control (3.9%). Several hematological
measurements were significantly different in both the warm and cold selenium
exposures relative to controls. Both warm and cold selenium treatments also had
greater O2 consumption than controls. Fish lipid content in the cold Se treatment
decreased more than the cold control; lipid content did not decrease in either the
warm control or the warm Se treatment (see summary tables below). The results
suggest significant mortality occurs in juvenile bluegill during winter months
when tissue concentrations reach 7.91 mg/kg dw and lipid levels decrease to 6
percent.
20°C, >6 mg Se/kg whole-body; 4°C, <7.91 mg Se/kg dw whole body
See "Comparison of the Cold-Temperature Bluegill Juvenile-Survival Studies" in
this appendix after presentation of the Mclntyre et al. (2008) study.
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Mean Concentration of Selenium in Tissues, Cumulative Survival*, Percent Lipid Content and Oxygen Consumption in Juvenile
Bluegill
day
0
60
120
180
cold - Se control
Sea
1
1
1.1
1.4
Surv.
%
100
97.1
97.1
97.1
lipid,
%
13.2
12.5
11.5
10.5
02b
98
58
57
57
cold + Se
Sea
1
5.8
7.9
7.9
Surv.
%
100
92.9
66.4
59.6
lipid,
%
13.2
10
6
6
02b
98
63
81
78
warm - Se control
Sea
1
1.2
1.1
1.2
Surv.
%
100
95.7
95.7
95.7
lipid,
%
13.2
13.3
13.4
13.6
02b
98
98
100
100
warm + Se
Sea
1
5.8
6
6
Surv.
%
100
100
96.7
92.6
lipid,
%
13.2
13.3
13.4
13.5
02b
98
103
120
120
a whole body Se tissue concentration, mg/kg dw
b oxygen consumption, mg/kg/hr
* Cumulative Survival: In this experiment, 240 juvenile bluegill were placed in three 400-L fiberglass tanks, 80 in each, and exposed to
each control and treatment for a period of 180 days. Ten fish were removed at random from each treatment replicate on days 0, 60, 120,
and 180 for selenium, histological, hematological, and metabolic measurements.
Replicate and Average Whole-body concentrations (mg/kg dry weight) of selenium in juvenile bluegill*
replicat
e
c+Se
w+Se
c-Se
w-Se
dayO
1
0.87
1.17
0.89
0.99
2
1.21
0.96
3
0.95
0.90
mean
1.01
1.01
0.89
0.99
day 60
1
6.30
5.61
0.97
1.12
2
5.49
6.19
3
5.76
5.43
mean
5.85
5.74
0.97
1.12
day 120
1
8.36
6.37
1.01
0.99
2
7.31
5.92
3
7.85
5.50
mean
7.84
5.93
1.01
0.99
day 180
1
7.53
5.48
1.10
0.96
2
8.01
5.72
3
8.19
6.02
mean
7.91
5.74
1.10
0.96
* Each value is for a composite sample made from 5 fish.
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C-149
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The Kaplan-Meier estimator was used to calculate survival at time t
5(0 =
where r(t,) is the number offish alive just before time tt, i.e. the number at risk, and dt is the number of deaths in the interval It = \tt, ti+i\.
The 95% confidence interval for such estimate (Venables and Ripley 2002) was computed as
expi-//(/)exp
±k.
H(t)
where
d.
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C-150
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The following table lists the estimates of survival in the cold + Se treatment at 60, 120 and 180 days. The term n.event is the number of
deaths at a given interval; n.risk is the number of organisms alive at the beginning of the interval; survival is computed by the Kaplan-
Meier estimator.
Time
60
120
180
n.risk
210
165
88
n. event
15
47
9
survival
0.929
0.664
0.596
std.err
0.0178
0.0350
0.0381
lower 95% CI
0.884
0.590
0.517
upper 95% CI
0.956
0.728
0.666
Hematological Measurements in Juvenile Bluegill Sunfish (indicates significantly different from control)
Warm Exposure
blood parameter
total erythrocyte, 106/ml
% mature
nuclear shadows, 104/ml
total leucocytes, 104/ml
% lymphocytes
% neutrophils
hematocrit, %
MCHC (mean corpuscular hemoglobin
cone.)
Col d Exposure
blood parameter
dayO
warm-Se
2.95
85
0.95
17.22
23
15
37
23
warm+Se
2.92
86
0.86
17.41
25
13
36
25
dayO
cold-Se
cold+Se
day 60
warm-Se
2.96
86
0.97
16.90
20
14
37
25
warm+Se
2.93
93*
2.05*
17.55
23
15
29*
19*
day 60
cold-Se
cold+Se
day 120
warm-Se
2.99
86
0.83
16.73
19
17
36
25
warm+Se
2.95
94*
2.38*
17.62
26
19
29*
18*
day 120
cold-Se
cold+Se
day 180
warm-Se
2.96
85
0.91
17.05
21
17
38
25
warm+Se
2.89
94*
2.30*
17.36
22
16
28*
17*
day 180
cold-Se
cold+Se
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C-151
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total erythrocyte, 106/ml
% mature
nuclear shadows, 104/ml
total leucocytes, 104/ml
% lymphocytes
% neutrophils
hematocrit, %
MCHC (mean corpuscular hemoglobin
cone.)
MCV (mean corpuscular volume)
2.91
84
0.86
16.48
17
13
39
26
182
2.93
82
0.84
16.88
16
12
37
25
171
2.97
87
0.83
16.79
16
15
40
25
188
2.90
95*
2.30*
16.91
17
11
30*
18*
146*
3.01
85
0.89
16.80
19
15
41
22
180
2.95
96*
2.49*
16.74
15
12
28*
17*
135*
3.00
85
0.90
16.96
19
12
39
23
185
2.99
97*
2.36
16.63
18
14
27*
17*
130*
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Mclntyre et al. 2008. Effect of Selenium on Juvenile Bluegill Sunfish at Reduced Temperatures. US
EPA, Health and Ecological Criteria Division. EPA-822-R-08-020
Test Organism:
Exposure Route:
Test Duration:
Study Design:
Bluegill sunfish (Lepomis macrochirus); juvenile; average length 47 mm,
average weight 1 g
Waterborne and dietary
Water
1:1 selenite:selenate; For exposure systems (ES) 1 and 3, fish were exposed to a
control and a series of 6 nominal concentrations, 1.25, 2.5, 5, 10, 20 and 40 (ig
Se/L. For ES2, fish were exposed to a control and one nominal concentration, 5
(ig Se/L.
Diet
For ESI and ESS, fish were fed a series of six concentrations of selenium and a
background control mLumbriculus variegatus. The measured selenium
concentrations in the L. variegatus treatments in ESI were: 2.3 (control), 4.5, 5.3,
7.5, 14.2, 25.7 and 34.9 mg Se/kg dw; in ES3: 2.2 (control), 4.2, 5.0, 7.2, 15.2,
25.4 and 46.7 mg Se/kg dw. Fish were fed worms at a rate of 4% of the current
biomass in each fish tank. Selenium was accumulated in L. variegatus by feeding
the worms in separate tanks a series of six concentrations of selenized-yeast
diluted with nutritional yeast: 1.7, 3.3, 6.7, 13.3, 26.7 and 53.5 mg Se/kg dw.
Control worms were fed nutritional yeast only. Each tank was additionally
exposed to the associated aqueous concentration selenium, e.g., the worms fed
the 1.7 mg Se/kg dw selenized yeast were exposed to 1.25 :g Se/L, the worms fed
the 3.3 mg Se/kg dw selenized yeast were exposed to 2.5 :g Se/L, and so on.
For ES2, fish were fed TetraMin spiked with seleno-L-methionine at a nominal
concentration of 5 mg/kg dw and at a rate of 3% of the current biomass in each
tank.
182 days
Juvenile bluegill were exposed concurrently to selenium using three separate
exposure systems, ESI, ES2 and ES3. In ESI and ES3, 100 fish were exposed to
each of 6 selenium treatments (low through high treatments are referred to as
Treatments 1 through 6) and two controls in 200 L carboys under flow-through
conditions. Each treatment consisted of an aqueous selenium concentration and
an associated dietary selenium concentration, e.g., the fish in the lowest ESI
treatment were exposed to 1.25 :g Se/L and fed worms containing 4.5 mg Se/kg
dw (see Exposure Route for other treatment concentrations). Temperature was
controlled in each system through the immersion of the carboys in a temperature-
controlled water bath and by controlling the temperature of the dilution water
being added to the carboys. The temperature in ESI was maintained at 20°C for
the first 30 days of exposure, and then decreased 2°C/week until it reached 4°C
(test day 79) at which point temperature was maintained until test termination
(test day 182). The only difference between ESI and ES3 was temperature was
decreased 2°C/week until it reached 9°C (test day 65) at which point temperature
was maintained until test termination (test day 182).
The exposure of ES2 was similar to ESI and ES3 in that 100 juvenile bluegill
were exposed to treatment in 200 L carboys under flow-through conditions. The
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ES2 selenium treatment consisted of two replicates of 5 (ig Se/L waterborne and
5 mg Se/kg dw diet (Tetramin). Two controls were maintained with ES2. The
temperature regime for ES2 was identical to ES1.
Observations on fish behavior and mortality were checked daily. Total selenium
was measured in each fish tank weekly and selenium speciation was measured
monthly in each fish tank. Whole body total selenium was measured in the
worms from each tank (2 replicate 5 g samples) on test days 0, 30, 60, 112 and
182 and in the bluegill from each tank (3 replicates of 3-fish composites - total 9
fish) on test days 0, 7, 30, 60, 112 and 182. The standard length and weight of
each fish was measured on each sample day. Lipid content was measured in fish
at day 0 and from each treatment at test termination.
Effects Data: Selenium increased in bluegill as the exposure concentrations increased (see
following table). No meaningful mortality was observed in ES2. The number of
fish that died in ES2 during the 182 day test were two fish in one treatment
replicate and none in the other treatment replicate; no deaths were reported in
ES2 controls. Significant mortality of juvenile bluegill was observed in ESI and
ES3. After 182 days, a total of 24 and 68 fish died in Treatments 5 and 6,
respectively in ESI; and a total of 38 and 61 fish died in Treatments 5 and 6,
respectively in ES3. See table below for mortalities in all treatments. Estimates of
bluegill survival were adjusted for the removal of individuals from the test
population. Individuals were removed from the experiments before test
completion, for sampling tissue concentrations or because they suffered
accidental deaths unrelated to selenium toxicity. For such data, it was necessary
to account for the reduction in number of individuals at risk of death due to
selenium over time. lfr(t,) is the number of individuals at risk just before time tt
and di is the number of deaths in the interval, /, = [th ti+l), then survival (S) at time
t can be estimated as
The product (P) was calculated for each period in which one or more deaths
occur. The equation is the Kaplan-Meier estimator (Venables and Ripley 2002).
This correction was applied to calculate the proportion of survival in treatments
with ten or more deaths (10% mortality). The table below provides the adjusted
proportion and surviving bluegill in each treatment along with the concentration
of selenium in bluegill at test termination. The values in this table were used to
calculate the EC20 and ECi0 values using the TEAM software. Growth and lipid
content of the bluegill was not negatively affected by the selenium exposures.
Do not distribute, quote or cite C-154 Draft Document
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Measured total selenium concentrations in bluegill sunfish for all treatments and controls in Exposure System 1, 2 and 3.
ESI
ES3
ES2
Control
Total Selenium in Whole Body Bluegill Tissue, mg/kg dw
Treatment 1 Treatment 2 Treatment 3 Treatment 4
Average (SD)
1.93(0.21)
2.48(0.11)
2.85(0.10)
2.70 (0.20)
3.16(0.11)
2.56(0.21)
Average (SD)
1.93(0.21)
2.43 (0.18)
3.10(0.04)
3.07(0.05)
3.41 (0.08)
3.15(0.25)
Average (SD)
1.93(0.21)
2.64 (0.06)
2.94(0.13)
3.69(0.25)
3.99(0.26)
4.02(0.21)
Average (SD)
1.93(0.21)
2.72 (0.07)
4.24 (0.22)
5.21 (0.30)
6.42 (0.05)
6.72 (0.09)
Treatment 5
Average (SD)
1.93(0.21)
3.27(0.27)
6.62 (0.23)
8.62 (0.45)
11.60(0.43)
10.71 (0.55)
Treatment 6
Average
(SD)
1.93(0.21)
4.27 (0.44)
10.21 (0.36)
12.66 (0.45)
Test Day
0
7
30
60
112
182
Test Day
0
7
30
60
112
182
Control
Average (SD)
1.93(0.21)
2.50(0.10)
2.24(0.41)
2.70 (0.22)
2.16(0.14)
1.67(0.21)
Control
Average (SD)
1.93(0.21)
2.19(0.19)
2.49(0.15)
1.53(0.03)
1.57(0.01)
1.38(0.06)
Treatment 1
Average (SD)
1.93(0.21)
2.60 (0.29)
2.44 (0.26)
2.88 (0.08)
2.49(0.10)
3.20(0.27)
5A
Average (SD)
1.93(0.21)
3.55 (0.25)
7.05 (0.76)
8.23(1.55)
8.97(1.28)
9.41(1.63)
Treatment 2
Average (SD)
1.93(0.21)
2.38(0.10)
2.70(0.16)
3.04(0.39)
3.10(0.12)
3.83 (0.47)
5B
Average (SD)
1.93(0.21)
3.08 (0.50)
7.51(1.18)
8.09 (0.67)
9.45(1.73)
10.61 (0.38)
Treatment 3
Average (SD)
1.93(0.21)
2.82 (0.20)
3.13(0.10)
3.79(0.24)
3.64(0.16)
5.48 (0.24)
Treatment 4
Average (SD)
1.93(0.21)
3.19(0.33)
3.95(0.16)
5.54(0.21)
6.54(0.21)
9.38 (0.63)
Treatment 5
Average (SD)
1.93(0.21)
4.29 (0.20)
6.06 (0.36)
9.50(0.91)
11.50(0.25)
16.01 (0.30)
Treatment 6
Average
(SD)
1.93(0.21)
6.13(0.62)
11.07(0.92)
15.14(0.96)
17.24 (0.30)
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Total number of deaths in ESI and ES3 Treatments throughout the experiment's duration (182
days). Both ESI and ES3 had two control tanks.
Treatment ES1 ESS
Control (#1, #2) 0,7 1,1
1 5 0
2 1 1
3 00
4 33
5 24 38
6 68 61
The concentration of selenium in bluegill and the adjusted proportion of surviving fish at the end of
the 182 day exposure.
ESI ES3
Treatment [Se]tiSSUe, rng/kg dw surv
control 2.08 0.962
1 2.56 0.988
2 3.15 0.984
3 4.02 1.000
4 6.72 0.962
5 10.71 0.497
6 12.66 0.075
[Se]tiSSue, mg/kg dw
1.67
3.20
3.83
5.48
9.38
16.01
17.24
surv
0.988
1.000
0.988
1.000
0.960
0.435
0.168
Chronic Value:
The NOAEC for bluegill in ES2 was calculated as the geometric mean of the
concentration of bluegill in the two replicates at the end of the exposure period,
9.992 mg Se/kg dw whole body. The chronic value for ES2 is therefore >9.992
mg Se/kg dw whole body. The EC2o and ECio values for ESI and ES3 are given
in the following table.
EC2o mg Se/kg dw
ECio mg Se/kg dw
ESI (4°C)
Whole body
9.78
9.27
ES3 (9°C)
Whole body
14.64
14.00
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Comparison of the Cold-Temperature Bluegill Juvenile-Survival Studies of Lemly (1993a) and
Mclntyre et al. (2008)
The Lemly (1993a) and Mclntyre et al. (2008) cold-temperature juvenile bluegill studies are summarized
on the previous pages. This discussion compares and contrasts these studies.
Both studies indicated that juvenile bluegill are more sensitive to selenium at lower temperature than at
higher temperature. For a 4°C temperature regime, the EC 10 of 9.27 mg Se/kg dw WB obtained with
Mclntyre's selenized yeast-worm-fish dietary bioaccumulation system is somewhat similar to the
threshold of 5.85 mg Se/kg dw WB estimated from the time course of bioaccumulation and mortality in
Lemly's single treatment with seleno-L-methionine in TetraMin. These chronic values differ by a factor
of 1.58.
The difference in diet does not appear to explain the modest difference in results; however, since
Mclntyre's other 4°C experiment (Exposure System ES2), which used Lemly's seleno-L-methionine in
TetraMin diet, experienced no significant toxicity, whereas Lemly's similarly exposed fish experienced
40 percent mortality by the end of the test. In addition to the difference in observed mortalities, Lemly=s
bluegill in the 4°C selenium exposure decreased in both lipid content and body condition over the 180
days whereas no decreases in these measurements were observed in the Mclntyre et al. study, although
the fish used in both studies were of comparable size and body condition at test initiation: 47 mm average
standard length (range 44 to 54 mm) and a body condition index (100 x fish weight/standard length) of
3.2 in ES2 compared to 50 to 70 mm total length and a body condition factor of 3.9 in Lemly.
There are several possible reasons why such results could differ between studies. (1) ES2 maintained
exposure at 20°C for the first 30 days of exposure before decreasing the temperature compared to 7 days
in the Lemly study. (2) Lemly measured O2 consumption by removing and reintroducing test fish to the
test tanks, which was not done by Mclntyre et al. (3) The two studies differed in photoperiod - Lemly
"began with a 16:10 h light/dark photoperiod which was gradually reversed to 10:16" (sic) whereas
Mclntyre et al. used a fixed photoperiod of 16:8. (4) Some genetic differences between the tested batches
of organisms may be expected, reflecting different origins, despite the similarities in their starting size
and condition.
The modification to maintain 20°C for 30 days was to allow a longer period of time for the fish to
accumulate selenium during a warmer condition prior to decreasing the temperature. This did result in
shortening the exposure in ES2 at 4°C by 19 days (103 days at 4°C) compared to 122 days at 4°C in
Lemly's study. However, as the majority of deaths in Lemly's study occurred between in the middle 60
days of the 180-day test, the slightly shorter cold period in the Mclntyre study would not explain the
differences in mortalities.
As stated above, Lemly removed fish (N = 15) from each treatment for oxygen consumption
measurement and then returned these fish to the exposure tanks. There is the possibility that the fish
removed from the cold plus selenium treatment were sufficiently stressed by the exposure conditions that
the additional handling stress contributed to the mortality observed in this treatment. Between test days 60
and 180, 56 fish died Lemly's cold plus selenium treatment. Even if stress due to handling affected all the
fish used in the oxygen consumption measurements (up to 30 fish), it does not explain all the mortality
that was observed and therefore does not explain the difference between the two studies.
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It may then be questioned whether the fixed photoperiod alone could account for the differences in the
results of the two studies. More explicitly, did the longer light period in Mclntyre et al. photoperiod allow
the fish to feed more than the fish exposed to the shorter light period in the Lemly study, such that lipid
and body condition in the Mclntyre et al. fish were maintained and therefore not susceptible to "winter
stress syndrome." The effects of photoperiod on fish and other ectotherms are well-documented.
Temperature-independent seasonal changes in fish have been reported for growth and food conversion
efficiency (Biswas and Takeuchi 2003; Jonassen et al. 2000; Simensen et al. 2000), feeding behavior
(Volkoff and Peter 2006), metabolic rate (Evans 1984), and reproduction (Koger et al. 1999; Scott 1979).
Some of these studies have found conflicting results on the effect of photoperiod on growth (Fuchs 1978;
Jonassen et al. 2000; Simensen et al. 2000). Coupled with temperature being a dominant factor in
controlling physiological functions in temperate-zone fish as indicated by a 3 to 4-fold fluctuation in
metabolic activities over 10°C (Brett 1970; Fry 1971), it is difficult to use literature findings to explain
the difference in the two bluegill studies.
Observational recordings of the feeding behavior in Mclntyre et al. noted that in both control replicates
and in both treatment replicates the feeding of the juvenile bluegill went from active to not active on test
day 78 when temperatures were decreased from 6.6 to 5.8°C. The feeding observations are reflected in a
gradual slight decrease in the body condition factor (K) after test day 60 in the figure below. Although
food intake was not quantified during the study, the lack of growth indicated in K suggests feeding
markedly decreased as the temperature declined, as shown in the figure. Body condition decreased much
more in the Lemly's cold plus selenium exposed fish after test day 60 (approximately 50%) but K in his
cold-without-selenium exposure decreased only slightly, similar to Mclntyre et al. Therefore it is not
possible to determine if the greater decrease in K and in lipid content in Lemly's cold plus selenium
treatment was due to decreased feeding because of a shorter photoperiod or because the bluegill fish
population used in his study were more sensitive to selenium in cold conditions. Mclntyre et al. obtained
bluegill from Osage Catfisheries in Missouri whereas Lemly collected fish from ponds (assumed to be
near Blacksburg, Virginia, not stated in paper). The fish obtained from Missouri, a location with colder
winters than Virginia, may have been better adapted for withstanding colder winter temperatures than
Lemly's fish and therefore were less sensitive to "winter stress syndrome" as induced by selenium
exposure. Similarly, different populations of a species can have varying sensitivities to stressors.
Furthermore, the relative difference in the Lemly and Mclntyre et al. results is slightly lower than Delos
(2001) found to be typical when equivalent toxicity tests of the same species are compared. There should
thus be no expectation that the two study results should agree more closely than they do.
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o
"G
CO
LL
c
g
'•^
T3
C
O
O
5 -
4 -
3 -
2 -
1 -
• Condition Factor (K)
Temperature (°C)
22
- 20
- 18
- 16
- 14
- 12
- 10
- 8
- 6
- 4
o
(D
50
100
150
200
Test Day
Relationship between body condition factor (K) and temperature in juvenile bluegill fed a diet of Se-
enriched TetraMin in the Mclntyre et al. (2008) study.
Both Lemly (1993) and Mclntyre et al. (2008) showed reduced survival of juvenile bluegill exposed to
elevated selenium under lab-simulated winter conditions, albeit at somewhat different concentrations. But
only Lemly, not Mclntyre et al., found the decreased survival to be accompanied by loss of lipid and body
condition. Such loss is not generally corroborated by field evidence (Janz 2008). Several studies have
measured growth and energy storage indicators in juvenile fish just prior to and just after winter at
reference sites and sites with elevated selenium in northern Canada (Bennett and Janz 2007a, b; Kelly and
Janz 2008; Driedger et al 2009; Weber et al. 2008). The growth (length, weight, condition factor, muscle
RNA:DNA ratio, muscle protein) and energy storage (whole body lipids, whole body triglycerides, liver
triglycerides, liver glycogen) indicators for five fish species (northern pike, burbot, fathead minnow,
creek chub, white sucker) measured just after winter were similar or greater than those measured just
before winter at the selenium exposed sites. The slimy sculpin did show a decrease in whole body
triglycerides, but the reduction was similar at exposed and reference sites.
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Carolina Power & Light. 1997. Largemouth Bass Selenium Bioassay- Report. Carolina Power & Light
Company, Environmental Services Section, 3932 New Hill, North Carolina. December 1997
Test Organism:
Exposure Route:
Test duration:
Study Design:
Effects Data:
Largemouth bass (Micropterus salmoides)
Laboratory; dietary exposure only; DL-selenomethionine added to an artificial
diet. Adult largemouth bass obtained from a commercial supplier were fed
several months prior to spawning a series of selenium concentrations in the
artificial diet.
Embryo-larval monitoring through swim-up stage.
Dietary exposure studies were conducted in 1995 and in 1996. In 1995, the
measured dietary Se concentrations were 0.9 (control), 2.9, 7.5 and 11.2 mg
Se/kg dw: in 1996, they were 26.7, 53.1 and 78.4 mg Se/kg dw. Parent fish were
fed to satiation twice per day. Approximately 100 eggs from each spawn were
transferred to each of 2 to 4 incubation cups. Eggs and larvae were monitored for
mortality and deformities up to the larval swim-up stage. Selenium was measured
in the liver, muscle and gonad tissues of the parent fish. All live deformed larvae
at swim-up stage were considered as mortalities in the analyses.
Over the two year period, 56 successful spawns were obtained across all dietary
treatments. Live larval fish with deformities (kyphosis, scoliosis, jaw gap, and
lordosis) and edema at swim-up stage were considered mortalities for data
analysis. The average concentration of selenium in ovaries ranged from 3.1
mg/kg dw in the control to 77.6 mg/kg dw in the high dietary treatment (Table 1).
Larval survival generally decreased as the selenium concentration in the ovary
increased (Table 1; Figure 1). A plot of the percent survival of larval largemouth
bass as a function of the logarithm of selenium concentration in the parental
female ovary using TRAP produced an EC10 of 20.35 mg Se/kg dw (Figure 1).
Effect
Concentration:
20.35 mg/kg dw in ovaries
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Table 1. Selenium concentrations in the diet, ovary and muscle tissues and the percent mortality
and deformities.
Measured Se in
diet fed to parents,
mg/kg dwa
0.9 ±0.1
(0.7- 1.3)
2.9 ±0.5
(2.1-3.8)
7.5 ±0.6
(6.3 - 8.4)
11.2±1.4
(9.3-14.1)
26.7 ± 1.7
(23.6-29.5)
53.1±4.8
(45.5-61.9)
Spawn No.
6
12
13
26
34
35
3
4
10 (2F)
13
14
9
12
15
18
1
2
5
7
8
16
19
6
11
17
2
5
11
16
17
19
36
37
51
52
22
25
30
31
32
41
48 (2F)
50 (2F)
55
Se in parent ovary, mg/kg dw
Individual
5.38
7.34
3.51
5.74
1.58
1.36
2.09
1.85
2.11
1.86
1.40
9.59
8.03
9.73
7.66
8.43
25.15
15.31
1.20
6.78
8.25
10.20
35.44
15.08
24.59
37.14
44.67
34.26
35.58
33.48
48.24
35.81
37.88
32.95
59.89
46.22
70.45
81.62
54.99
53.96
51.48
84.31
32.87
73.33
Average
3.1
8.8
10.8
25.0
40.0
61.0
Larval survival, %
Individual
75.5
99.7
96.2
88.9
99.5
96.8
98.8
100
97
97.1
98.4
84.9
100
98.5
95.9
75
63.9
90.6
79.1
95
96.8
100
91.5
77.9
96.7
91.2
0
75.9
0
9.9
0
6.3
0
0
0
0
0
0
0
0
0
0
0
0
Average
95.3
94.8
85.8
88.7
18.3
0
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Measured Se in
diet fed to parents,
mg/kg dwa
78.4 ±4.3
(73.2-87.0)
Spawn No.
4(2F)
7
8
10
18
21
24
28
38
44
47
49
Se in parent ovary, mg/kg dw
Individual
66.81
56.98
86.49
65.99
72.35
71.89
62.44
99.02
52.37
102.82
88.15
105.29
Average
77.6
Larval survival, %
Individual
66
0
0
0
0
0
0
0
0
0
0
0
Average
5.5
± standard error; range of concentrations in parentheses.
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Figure 1. Percent larval survival as a function of the logarithm of the selenium concentration
in the parental ovaries.
1MB all data
5 60 -
_ro 40 .
20 -
.5 1.0 1.5 2.0
Log(0vary Se, mg/kg dw)
Parameter Summary (Logistic Equation Regression Analysis!
Parameter Guess FinalEst StdError 95%LCL 95%UCL
LogXSO 1.4000 1.4828 0.0309 1.4207 1.5449
S 6.000 3.154 1.138 0.872 5.435
YO 97.00 93,37 4.44 84.46 102.28
Effect Concentration Summary
% Effect
50.0
20.0
10.0
5.0
Xp Est
30.40
23.60
20.35
17.758
95%LCL
26.35
17.65
13.77
10.920
95%UCL
35.07
31.56
30.09
28.876
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APPENDIX D: SUMMARY STUDIES OF NON-
REPRODUCTIVE EFFECTS
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STUDIES OF NON-REPRODUCTIVE EFFECTS
Acipenseridae
Acipenser transmontanus (white sturgeon)
Juvenile white sturgeon were exposed for 8 weeks to a series of 5 concentrations of seleno-L-methionine
added to an artificial diet (Tashjian et al. 2006). Survival was not affected by selenium treatment with a
mean survival rate of 99% across all groups. Fish fed the highest three dietary treatments of selenium,
41.7, 89.8 and 191.1 mg Se/kg dw, exhibited significant declines in growth assessed by body weight
measurements. The ECio for reduction in body weight is 15.08 mg Se/kg dw in whole body or 27.76 mg
Se/kg dw muscle; the EC2o is 17.82 mg Se/kg dw in whole body or 32.53 mg Se/kg dw muscle tissue. The
criterion values derived in this document that are based on reproductive endpoints are protective of the
endpoint measured in this non-reproductive study.
Cyprinidae
Pogonichthys macrolepidotus (Sacramento splittail)
Teh et al. (2004) exposed juvenile Sacramento splittail (7 months-old) to 8 levels of dietary selenium, 0.4
(no added selenium), 0.7, 1.4, 2.7, 6.6, 12.6, 26.0, and 57.6 mg/kg. Selenium was added to the diet via
selenized yeast which was diluted with Torula yeast (inactive) to attain the target levels. Mortality,
growth, histopathology, deformities and selenium content in muscle and liver were observed or measured
after 5 and 9 months of exposure. The appearance of deformities was the most sensitive endpoint. The
authors determined the occurrence of deformities was higher in fish fed 6.6 and 12.6 mg Se/kg in their
diet; however, such pathology was examined for only 15 of the 120 individuals per treatment, and a
consistent concentration-response relationship did not occur (i.e., no deformities in the high
concentration). The lack of a concentration-response relationship for the incidence of deformities has also
been observed in another study. Crane et al. (1992) exposed a European species of perch, Percafluviatilis
to three aqueous and dietary selenium treatments in experimental ponds for 288 days up through
spawning. Crane et al. (1992) found an increased occurrence of deformities in embryos and larvae in the
lowest selenium treatment relative to the control, but a decrease in the middle treatment. No hatching
occurred in the high treatment. Teh et al. (2004) proposed several physiological mechanisms to explain
the lack of a dose-response relationship, but it appears that the underlying mechanism is not understood at
this time. Toxicity tests with unusual dose-response relationships are typically not considered for criteria
derivation, but since another assay (Crane et al. 1992) observed a similar relationship, the Teh et al.
(2004) study with/1, macrolepidotus is included. Using prevalence of deformities as the endpoint, the
NOEC, LOEC and MATC (chronic value) in muscle tissue are 10.1, 15.1 and 12.34 mg Se/kg dw,
respectively. The critieron value in muscle tissue, based on the reproductive ECi0, is 11.8 mg Se/kg dw.
Appendix C provides further details on the study results and an approximate estimate of their relationship
to egg-ovary and whole-body concentrations. Teh et al. (2004) is the only study in which deformities
developed in fish that were not exposed to selenium from their mothers' ovaries. The selenium criterion
values derived based on reproductive endpoints are protective of the endpoint measured in this non-
reproductive study, considering the non-reproductive muscle MATC of 12.3 mg Se/kg dw is greater than
the reproductive muscle criterion of 11.8 mgSe/kg dw.
Pimephalespromelas (fathead minnows)
Non-reproductive chronic values for fathead minnows were derived from two laboratory-based studies.
These studies (Bennett et al. 1986 and Dobbs et al. 1996) involved exposing algae to selenium (either as
sodium selenite or sodium selenate) in water, and subsequently feeding the algae to rotifers which were in
turn fed to fathead minnows. In the Bennett et al. (1986) study, larval fathead minnows were fed control
rotifers (cultured in chambers without selenium containing algae) or selenium-contaminated rotifers
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(cultured in chambers with selenium containing algae previously exposed to sodium selenite in the water)
in three separate experiments lasting 9 to 30 days. The different experiments were distinguished by 1) the
day selenium-laden rotifers were first fed; 2) the day selenium-laden rotifers were last fed; and 3) the age
of larvae at experiment termination. The results from the three experiments reported by Bennett et al.
(1986) were conflicting. Larval growth was significantly reduced at larval whole-body selenium
concentrations of 43.0 mg Se/kg dw in the first experiment and 51.7 mg Se/kg dw in the second
experiment, but was slightly but not significantly reduced at 61.1 mg Se/kg dw in the third experiment
(see Appendix C). Following the approach of Section 7.1.1, the geometric mean of these three values,
51.40 mg Se/kg dw, is the chronic value for this study.
Dobbs et al. (1996) used a test system similar to that of Bennett et al (1986) (described above). Larval
fathead minnows were exposed to the same concentrations of sodium selenate in the water as their prey
(rotifers), but also received additional selenium from the consumption of the selenium-contaminated
rotifers. In this study, the fathead minnows did not grow well at concentrations exceeding 108.1 (ig Se/L
in water, and they survived only to 11 days at selenium concentrations equal to or greater than 393.0 (ig/L
in the water (75 mg Se/kg dw in the diet, i.e., rotifers). The LOEC for retarded growth (larval fish dry
weight) in this study was <73 mg Se/kg dw tissue.
A third laboratory study, by Ogle and Knight (1989), examined the chronic effects of elevated foodborne
selenium on growth and reproduction of fathead minnows. Juvenile fathead minnows were fed a purified
diet mix spiked with inorganic and organic selenium in the following percentages: 25 percent selenate, 50
percent selenite, and 25 percent seleno-L-methionine. The pre-spawning exposure lasted 105 days using
progeny of adult fathead minnows originally obtained from the Columbia National Fishery Research
Laboratory, as well as those obtained from a commercial fish supplier. After the 105 day exposure period,
a single male and female pair from each of the respective treatment replicates were isolated and inspected
for spawning activity for 30 days following the first spawning event of that pair. There was no effect from
selenium on any of the reproductive parameters measured, including larval survival, at the dietary
concentrations tested (5.2 to 29.5 mg Se/kg dw food). Sub-samples of larvae from each brood were
maintained for 14 days post-hatch and exhibited >87.4 percent survival. The pre-spawning adult fish fed a
mean dietary level of 20.3 mg Se/kg dw exhibited a significant reduction in growth compared to controls
(16 percent reduction), whereas a nonsignificant reduction in growth (7 percent) occurred in the fish fed
15.2 mg Se/kg dw. The chronic value, as determined by the geometric mean of the NOEC and the LOEC
measured at 98 days post-test initiation, was 17.57 mg Se/kg expressed as the above dietary
concentrations, and 5.961 mg Se/kg dw as fathead minnow whole-body tissue. The concentration-
response relationship, as indicated by the study data presented in Appendix E, was uniformly shallow; not
resembling the sharp sigmoidal function characteristic of most selenium response curves.
Since Ogle and Knight reported that food in the higher selenium concentrations remained uneaten and
fish were observed to reject the food containing the higher selenium concentrations, the authors suggested
that the decreased growth was caused by a reduced palatability of the seleniferous food items, which
contained unnatural percentages of inorganic selenium (Fan et al. 2002). This is a common observation
also noted by Hilton and Hodson (1983) and Flilton et al. (1980) and apparent in Coughlan and Velte
(1989). It is here interpreted to be an artifact of unrealistic spiking of the diet with inorganic selenium in
this early experimental protocol. That is, in the real world it is not expected that avoidance of food items
that were unpalatable because of excessive selenium would be either a mechanism by which selenium
causes effects or a mechanism by which organisms can avoid exposure. (See Janz et al. (2010) for a more
complete discussion of selenium's mechanism of toxicity.) Given the no observed effect on larval
survival and the apparent non-toxicological effect on growth in the Ogle and Knight study, a chronic
value for this study is not included.
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Catostomidae
Xyrauchen texanus (razorback sucker)
Two non-reproductive endpoint studies have been done with the endangered razorback sucker. In the first
study, Beyers and Sodergren (200 la) exposed larval razorback suckers for 28 days to a range of aqueous
selenate concentrations (6.12, 25.4, 50.6, 98.9, and 190.6 ug/L) and respectively fed them a range of
selenium in their diet (rotifers containing <0.702, 1.35, 2.02, 4.63, and 8.24 mg/kg dw). Reflecting the
lack of effects on survival and growth in any exposure, the chronic value for this study, based on selenium
measured in the larvae at the end of the test, is >12.9 mg Se/kg dw.
In a second study, Beyers and Sodergren (200 Ib) exposed larval razorback suckers to a control water and
three different site waters containing varying concentrations of selenium for 28 days. Two treatments
were tested within each water type: fish fed rotifers cultured in the same water type (site diet) and fish fed
rotifers cultured in control water. There were no reductions in survival or growth in fish exposed to both
the site water and site diet compared to fish exposed to control water and control diet. There were,
however, reductions in growth of fish exposed to site water/site food compared to the same site water and
control food. The authors did not attribute the effect on larval growth by the diet to selenium and cited
several lines of evidence, including: (1) there was not a dose-response relationship in the concentration of
selenium in the food (rotifers) and growth, nor in the concentration of selenium in the fish larvae and
growth across the three water types; and (2) water from the De Beque site promoted a significant
reduction in the growth of fish exposed to site water/site food relative to site water/control food, but
contained low levels of selenium in the water (<1 ug/L) and in food (2.10 mg/kg dw) typically lower than
those that have been found to elicit effects. The chronic value for this study is >42 mg Se/kg dw based on
the whole body concentration of selenium in the larval razorback suckers exposed to North Pond site
water.
Two similar studies were conducted in 1996 and 1997 to determine effects of site water and site food,
both contaminated with selenium, on the razorback sucker (Hamilton et al. 2001a,b; published later in a
peer-reviewed journal in 2005, see Hamilton et al. 2005 a,b,c). Both studies show marked effects of
selenium on survival of razorback sucker larvae exposed to contaminated food and to a lesser extent,
contaminated water. Although the data convincingly demonstrate effects to larval survival from exposure
to contaminated food, interpretation of the results, of chronic criterion derivation is complex because of
inconsistencies between: 1) levels of selenium in the food and larvae relative to larval survival; 2) the
time to larval mortality relative to selenium in the diet and selenium in the larvae; and 3) levels of other
inorganic contaminants in food and water (possible organic contaminants were not measured). Summaries
of each of these two studies as well as a third study with razorback suckers (Hamilton et al. 2005d) are
presented in Appendix E.
Due to the confounding results, lack of dose-response within and among related studies, and the
uncertainty of the effect of other inorganic contaminants on larval response to the various dietary and
waterborne treatments, the data from these three studies for razorback sucker (Hamilton et al. 2001a,b;
Hamilton et al. 2005d) have not been included. A more detailed explanation of why these studies were not
included is given in Appendix E. Because of the vastly different results between the Beyers and
Sodergren studies and Hamilton et al. studies and the inability to resolve the differences, SMCV and
GMCV were not calculated for the razorback sucker.
Catostomus latipinnis (flannelmouth sucker)
Beyers and Sodergren (200 la) exposed flannelmouth sucker larvae to a range of aqueous selenate
concentrations (<1, 25.4, 50.6, 98.9, and 190.6 ug/L) and fed them a range of selenium in their diet
(rotifers containing <0.702, 1.35, 2.02, 4.63, and 8.24 mg/kg dw, respectively). There were no survival or
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growth effects observed after the 28 day exposure. The chronic value based on the concentration of
selenium measured in the larvae exposed to the highest test concentration was >10.2 mg Se/kg dw.
Salmonidae
Oncorhynchus tshawytscha (Chinook salmon)
Hamilton et al. (1990) conducted a 90-day growth and survival study with swim-up larvae fed one of two
different diets. The first diet consisted of Oregon moist™ pellets where over half of the salmon meal was
replaced with meal from selenium-laden mosquitofish (Gambusia affinis) collected from the San Luis
Drain, CA (SLD diet). The second diet was prepared by replacing half the salmon meal in the Oregon
moist™ pellets with meal from low-selenium mosquitofish (i.e., the same relatively uncontaminated
mosquitofish that were used in the control diet) and spiked with seleno-DL-methionine (SeMe diet).
Analysis of the trace element composition in the two different diets indicated that while selenium was the
most toxic element in the SLD diet, concentrations of boron, chromium, iron and strontium in the high-
selenium mosquitofish replacement diet (SLD diet type) were slightly elevated compared to the
replacement diet. These trace elements were, however, only 1.2 (e.g., iron) to 2.0 times (e.g., chromium)
higher in the SLD diet than the SeMe diet, which contained the following measured concentrations (dry
weight basis) in the food: 10 mg boron/kg, 2.8 mg chromium/kg, 776 mg iron/kg, and 48.9 mg
strontium/kg.
During the test, survival of control Chinook salmon larvae (consuming food at approximately 3 mg Se/kg
dw) was 99 percent up to 60 days post-test initiation. Between 60 and 90 days of exposure, however, the
control survival declined to 66.7% in the SLD test and to 72.5% in the test using the SeMe diet, indicating
compromised health. Therefore, only data collected up to 60 days post-test initiation were considered for
analysis. Nevertheless, there remains the possibility that even at 60 days, the control organisms were not
healthy, although overt signs of stress did not appear until later.
For the SeMe diet, regression analysis of the 60-day growth data yielded a whole-body ECio of 7.355 mg
Se/kg dw and an EC2o of 10.47 mg Se/kg dw. For the SLD diet, regression analysis of the 60-day growth
data yielded a whole-body ECi0 of 11.14 mg Se/kg dw and an EC2o of 15.73 mg Se/kg dw. Note: The San
Luis Drain mosquitofish (comprising the Chinook salmon's SLD diet) were not tested for contaminants
other than certain key elements. Because the San Luis Drain receives irrigation drainage from the greater
San Joaquin Valley, there is a possibility that the SLD diet might have contained elevated levels of
pesticides, possibly a confounding factor, although the SLD diet was less toxic than the SeMe diet.
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Oncorhynchus mykiss (rainbow trout)
Hilton and Hodson (1983) reared juvenile rainbow trout on either a high (25 percent) or low (11 percent)
available carbohydrate diet supplemented with sodium selenite for 16 weeks. Body weights, feed: gain
ratios, and total mortalities were followed throughout the exposure every 28 days. Tissues (livers and
kidneys) were extracted for selenium analysis after 16 weeks. By the end of the exposure, fish fed diets
(low carbohydrate and high carbohydrate) with the highest selenium concentrations (11.4 and 11.8 mg
Se/kg dw food, respectively) exhibited a 45 to 48 percent reduction in body weight (expressed as kg per
100 fish) compared to control fish. The authors attributed such results to food avoidance. With only two
dietary exposure concentrations and a control, these data were not amenable to regression analysis. The
MATC for growth of juvenile rainbow trout relative to the concentrations of selenium in liver tissue of
trout reared on the high carbohydrate seleniferous dietary type is the geometric mean (GM) of 21.00 mg
Se/kg dw liver (NOEC) and 71.7 mg Se/kg dw liver (LOEC), or 38.80 mg Se/kg dw liver. The calculated
MATC for the same group of experimental fish exposed to selenium in the low carbohydrate diet is 43.5
mg Se/kg dw liver tissue, which is the same MATC for trout exposed for an additional 4 weeks based on
the occurrence of nephrocalcinosis in kidneys (see Hicks et al. 1984; Appendix C).
Hilton et al. (1980) employed a similar test design to that of Hilton and Hodson (1983) to examine the
narrow window at which selenium changes from an essential nutrient to a toxicant affecting juvenile
rainbow trout. The food consisted of a casein-Torula yeast diet supplemented with selenium as sodium
selenite. As discussed previously for the Ogle and Knight (1989) study with fathead minnow, this
represents an unrealistic fraction of inorganic selenium in the diet. The experiment lasted for 20 weeks.
During this time, the trout were fed to satiation 3 to 4 times per day, 6 days per week, with one feeding on
the seventh day. Organs (liver and kidney) and carcasses were analyzed for selenium from fish sacrificed
at 4 and 16 weeks. No gross histopathological or physiological effects were detected in the fish, although
trout raised on the highest dietary level of selenium (13.06 mg Se/kg dw food) had a significantly lower
body weight (wet basis), a higher feed:gain ratio, and higher number of mortalities (10.7; expressed as
number per 10,000 fish days). The MATC for growth and survival of juvenile rainbow trout relative to
the final concentrations of selenium in liver tissue is the geometric mean of the NOEC (40 mg Se/kg dw
liver) and the LOEC (100 mg Se/kg dw liver), or 63.25 mg Se/kg dw, both of which hinge on accepting
dietary spiking entirely with inorganic selenium as an acceptable experimental protocol.
The non-reproductive GMCV for Oncorhynchus (both rainbow trout and Chinook salmon) is 9.052 mg
Se/kg dw whole body based on the ECio value derived from the Hamilton et al. (1990) study with
Chinook salmon. The NOEC values for the rainbow trout studies conducted by Hilton and Hodson
(1983), Hilton et al. (1980), and Hicks et al. (1984) were not used in the GMCV calculation because of
the large difference between the NOEC and the LOEC values. If adult fish contained whole-body
selenium concentrations equal to 9.052 mg Se/kg dw, their egg-ovary concentrations would be estimated
to be 21.5 mg Se/kg dw when translated using the factor 2.37. The criterion values derived based on
reproductive endpoints are protective of the endpoint measured.
Moronidae
Morone saxitilis (striped bass)
A non-reproductive chronic value for selenium was determined from a laboratory dietary exposure
conducted using yearling striped bass (Coughlan and Velte 1989). During the experiment, the bass were
fed contaminated red shiners (38.6 mg Se/kg dw whole body) from Belews Lake, NC (treated fish) or
golden shiners with low levels of selenium (1.3 mg/kg dw whole body) purchased from a commercial
supplier (control fish). The test was conducted in soft well water and lasted up to 80 days. During the
experiment, all fish were fed to satiation 3 times per day. Control fish grew well and behaved normally.
Treated fish behaved lethargically, grew poorly due to a significant reduction in appetite, and showed
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histological damage, all eventually leading to the death of animals. The final selenium concentration in
muscle of treated striped bass averaged from 16.2 to 18.5 mg/kg dw tissue (assuming 78.4 percent
moisture content), which was 3.4 to 3.6 times higher than the final selenium concentrations in control
striped bass, which averaged 5.10 mg/kg dw tissue. The chronic value for this species was determined to
be <16.2 mg Se/kg dw in muscle tissue.
Centrarchidae
Lepomis macrochirus (bluegill)
Bryson et al. (1985b) conducted juvenile survival toxicity tests using hatchery bluegill and various forms
of selenium spiked to an artificial diet as well as a diet consisting of zooplankton collected from Hyco
Reservoir. There was no effect on length or weight of the juvenile bluegill after 60 days of exposure. The
highest concentration of selenium measured in whole body of the juveniles in these tests was in the
seleno-DL-cysteine-2X treatment (3.74 mg Se/kg dw).
Cleveland et al. (1993) performed a 90-day diet-only laboratory exposure in which juvenile bluegill were
fed a range of selenomethionine concentrations added to Oregon moist™ pellets. The authors observed no
significant effects on survival, but did report a very small but apparently statistically significant decrease
in the condition factor, K, from 1.3 at four concentrations between 1.0 and 4.7 mg Se/kg dw whole body,
to 1.2 at the two concentrations 7.7 and 13.4mg Se/kg dw whole body. The condition factor (weight x
105/length3) is intended to reflect a fish's reserves. In contrast to the studies of Ogle and Knight (1989),
Hilton and Hodson (1983), and Hilton et al. (1989), which appear to have involved an inorganic selenium
food palatability problem, this study did not use inorganic selenium in the diet. Nevertheless, given that
the reduction in K (1.3 to 1.2) is slight and shows no increasing effect between 7.7 and 13.4 mg Se/kg dw,
thus not yielding a sigmoidal concentration-response curve to support an ECio calculation, the chronic
value for this study was estimated at >13.4 mg Se/kg dw in whole body tissue.
Data from Lemly (1993a) indicate that over-wintering fish may be more susceptible to the effects of
waterborne and dietary selenium due to increased sensitivity at low temperature. The author exposed
juvenile bluegill in the laboratory to a single elevated exposure level, waterborne (1:1 selenite:selenate;
nominal 5 (ig Se/L) and foodborne (seleno-L-methionine in TetraMin; nominal 5 mg Se/kg dw food)
selenium for 180 days. Tests with a control and the treated fish were run at 4°C and 20°C with biological
and selenium measurements made every 60 days. Survival and whole-body lipid content were unaffected
at 20°C (whole-body selenium concentrations equal to 6 mg/kg dw, the sole treatment exposure) when
compared to control fish. Thus, at 20°C the chronic value for juvenile bluegill exposed to waterborne and
dietary selenium based on survival was >6 mg/kg dw in whole-body tissue. Fish exposed to the
combination low-level waterborne and dietary selenium at 4°C exhibited significantly elevated mortality
(40.4 percent) relative to controls (2.9 percent), and exhibited significantly greater oxygen consumption
and reduced lipid content, which are indicative of stress. At 4°C the chronic value for juvenile bluegill
exposed to waterborne and dietary selenium was <7.91 mg Se/kg dw in whole body based on mortality
and tissue measurements at the end of the test (180 days), and 5.85 mg Se/kg dw in whole body based on
mortality at 180 days and tissue measurements at 60 days. The increase in the concentration of whole-
body selenium between Day 60 and 180 at 4EC was apparently due to reductions in body weight caused
by loss of lipid (comparatively low in selenium) while body burden in other tissues remained relatively
constant. If this concentration of selenium in tissues occurs in sensitive overwintering fish in nature, a
concentration of 5.85 mg/kg dw (the selenium tissue concentration in the 4°C exposure after 60 days) in
fish collected during the summer or fall months could be considered a threshold concentration for the
selenium-sensitive fish during the winter months. Therefore, this study's chronic value for the threshold
concentration prior to winter stress is 5.85 mg Se/kg dw in whole body tissue.
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Mclntyre et al. (2008) also investigated the toxicity of selenium to juvenile bluegill under cold
temperature conditions in the laboratory. Whereas relative to the control, Lemly (1993a) tested only one
exposure level, 5 mg Se/kg in the diet and 5 ug Se/L and one low temperature regime, 4°C, Mclntyre et
al. (2008) evaluated a range of diet and water concentrations, two types of diet, and two low-temperature
regimes. The goal of the study was to determine ECio and EC2o values for selenium exposure to juvenile
bluegill in 4°C and 9°C low-temperature regimes. Three separate exposure systems were run concurrently
for 182 days. Two systems exposed juvenile bluegill to a series of six aqueous and dietary selenium
treatments and a control; one exposure system (ESI) with a cold temperature regime (4°C), and one (ESS)
with a cool temperature regime (9°C), both using a yeast-worm-fish food chain bioaccumulation system.
That is, graded levels of selenized-yeast in ESI and ESS were fed to the oligochaete, Lumbriculus
variegatus, which in turn was fed to bluegill. The third exposure system (ES2) used diet and exposure
conditions similar to Lemly's 4°C treatment, i.e., nominal 5 (ig Se/L in the water and nominal 5 mg Se/kg
dw food (seleno-L-methionine in TetraMin). The cold temperature regime for ESI and ES2 was 20°C for
the first 30 days of exposure, and then decreased 2°C/week until it reached 4°C (test day 79) at which
point temperature was maintained until test termination (test day 182). The cool temperature regime
(ESS) was similar except when the temperature reached 9°C (test day 65), it was maintained until test
termination (test day 182).
At the end of the 182 day exposure in the ES2 (with Lemly's diet and temperature), the bluegill
accumulated an average (geometric mean) whole body concentration of 9.99 mg/kg dw with no
meaningful mortality in the treatment or control. Significant mortality of juvenile bluegill was observed in
the two highest treatments in the cold (ESI) and cool (ESS) Lumbriculus-fed tests. No effects on body
weight or condition factor were observed. The ECi0 and EC20 values for the cold treatment (ESI) are 9.27
and 9.78 mg Se/kg dw in whole body, respectively. The ECi0 and EC2o values for the cool treatment
(ESS) are slightly higher at 14.00 and 14.64 mg Se/kg dw in whole body, respectively.
The design and the results of the Mclntyre et al. (2008) study have similarities and differences with
Lemly (1993a), as presented in detail with comparisons and contrasts in Appendix C. Both studies found
juvenile bluegill were more sensitive in a cold-temperature regime than in a cool (Mclntyre et al.) or a
warm regime (Lemly). The effect levels determined for the cold temperature regime differed by a factor
of 1.58 (ESI of Mclntyre et al., 9.27 mg Se/kg; Lemly, 5.85 mg Se/kg), a difference rather typical of
chronic studies conducted in different laboratories using different fish populations (Delos 2001) and
similar to the 1.51 factor difference between two ECi0s of Hamilton etal. (1990) for chinook salmon. As
these two cold-temperature juvenile-survival lab studies are far more similar than they are different, their
results were combined per the standard procedure described in the U.S.EPA Ambient Water Quality
Criteria Guidelines, to determine the non-reproductive SMCVs for bluegill. These SMCVs were
determined separately for two temperature conditions for bluegill, 4°C and 9°C. The SMCV for 4°C is
8.15 mg Se/kg dw whole body, based on three chronic values: (a) the Lemly (1993a) concentration prior
to winter stress (5.85 mg Se/kg dw whole body), (b) the Mclntyre et al. (2008) ESI EC10 (9.27 mg Se/kg
dw whole body), and (c) the Mclntyre et al. (2008) ES2 NOEC (>9.992 mg Se/kg dw whole body). This
value is not less than the reproductive endpoint-based whole-body criterion concentration of 8.0 mg Se/kg
dw. The SMCV for 9°C is 14.00 mg Se/kg dw whole body, based on the Mclntyre et al. (2008) ESS EC10.
The studies of Bryson et al (1985b) and Cleveland et al. (1993) were not conducted at cold temperatures
and were thus not used for these SMCV calculations.
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Table D-l (same as Table 17 in the main document). Freshwater Chronic Values from Acceptable Tests - Non-Reproductive
Endpoints (Parental Females Not Exposed).
Species
Acipenser
transmontanus
white sturgeon
Pogonichthys
macrolepidotus
Sacramento splittail
Pimephales promelas
fathead minnow
Pimephales promelas
fathead minnow
Xyrauchen texanus
razorback sucker
Xyrauchen texanus
razorback sucker
Catostomus latipinnis
flannelmouth sucker
Oncorhynchus
tshawytscha
Reference
Tashjian et al.
2006
Teh etal. 2004
Bennett etal. 1986
Dobbsetal. 1996
Beyers and
Sodegren2001a
Beyers and
Sodegren2001b
Beyers and
Sodegren2001a
Hamilton et al.
1990
Exposure route and
duration
dietary (lab)
8 weeks
dietary (lab)
9 months
dietary (lab)
9 to 19 days
dietary and
waterborne
(lab)
8 days
dietary and
waterborne (lab)
28 days
dietary and
waterborne (lab)
28 days
dietary and
waterborne (lab)
28 days
dietary (lab)
60 days
Selenium form
seleno-L-methionine in
artificial diet
seleno-L-methionine in
artificial diet
selenized-yeast
algae exposed to selenite
then fed to rotifers which
were fed to fish
algae exposed to selenate
in water then fed to
rotifers which were fed
to fish
water: selenate; diet:
algae exposed to selenate
in water then fed to
rotifers which were fed
to fish
water: site waters; diet:
algae exposed to site
water then fed to rotifers
which were fed to fish
water: selenate;
diet: algae exposed to
selenate in water then
fed to rotifers which
were fed to fish
mosquitofish spiked with
seleno -DL-methionine
lexicological
endpoint
ECiojuvenile
growth
EC20 juvenile
growth
NOEC
LOEC
MATC juvenile
deformities
(juvenile exposure
only)
Chronic value for
larval growth
LOEC for larval
fish dry weight after
8d
NOEC for survival
and growth
NOEC for survival
and growth
NOEC for survival
and growth
EC10 for juvenile
growth
Chronic value,
mg/kg dwa
15.08 WB
27.76 M
17.82 WB
32.53 M
10.1 M
15. 1M
12.34 M
51.40 WB
<73 WBb
>12.9WBb
>42 WBb
>10.2 WB
7.355 WB
SMCV
mg/kg dw
EC10
15.1 WB
27.8 M
EC20
17.8 WB
32.5 M
10.1 M
15. 1M
12.3 M
51.40WB
69.83 M
see text
>10.2 WB
EC10
9.052 WB
GMCV
mg/kg dw
15.1 WB
27.8 M
10.1 M
15. 1M
12.3 M
51.40WB
69.83 M
see text
>10.2 WB
EC10
9.052 WB
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Species
chinook salmon
Oncorhynchus mykiss
rainbow trout
Oncorhynchus mykiss
rainbow trout
Morone saxitilis
striped bass
Lepomis macrochirus
bluegill
Lepomis macrochirus
bluegill
Reference
Hilton and Hodson
1983;
Hicks etal. 1984
Hilton etal. 1980
Coughlan and
Velte 1989
Lemly 1993a
Mclntyre et al.
2008
Exposure route and
duration
dietary (lab)
16 weeks
dietary (lab)
20 weeks
dietary (lab)
80 days
dietary and
waterborne (lab)
180 days
20 to 4°C
dietary and
waterborne (lab)
180 days 20°C
dietary and
waterborne (lab)
182 days
20to4°C(ESl)
dietary and
waterborne (lab)
182 days
20 to 9°C (ESS)
dietary and
waterborne (lab)
182 days
20 to 4°C (ES2)
Selenium form
mosquitofish spiked with
SLD diet
sodium selenite in food
preparation
sodium selenite in food
preparation
Se-laden shiners from
Belews Lake, NC
diet: seleno-L-
methionine
water: 1:1
selenate: selenite
diet: seleno-L-
methionine
water: 1:1
selenate: selenite
diet: Lumbriculus fed
selenized-yeast
water: 1:1
selenate: selenite
diet: Lumbriculus fed
selenized-yeast
water: 1:1
selenate: selenite
diet: seleno-L-
methionine
water: 1:1
selenate: selenite
lexicological
endpoint
EC20 for juvenile
growth
EC10 for juvenile
growth
EC20 for juvenile
growth
juvenile growth
NOEC
LOEC
MATC
juvenile survival
and growth
NOEC
LOEC
MATC
LOEC for survival
of yearling bass
LOEC for juvenile
mortality at 4oC
Threshold prior to
"winter stress"
NOEC for juvenile
mortality at 20oC
EC10juv. survival
ESI
EC20juv. survival
ESI
ECiojuv. survival
ESS
EC2ojuv. survival
ESS
NOECjuv. surv.
ES2
Chronic value,
mg/kg dwa
10.47 WB
11.14WB
15.73 WB
21 Liver
7 1.7 Liver
38.80 Liver
40 Liver
100 Liver
63.25 Liver
<16.2Mc
<7.91 WB
5.85 WB
>6.0WB
9.27 WB
9.78 WB
14.00 WB
14.64 WB
>9.992 WB
SMCV
mg/kg dw
EC20
12.83 WB
NOAEC
28.98 L
LOAEC
84.68 L
MATC
49.52 L
<16.2M
4°C
Edo-NOAEC
8.15 WB
4°C
EC20-LOAEC
8.80 WB
9°CEC10
14.0 WB
9°C EC20
14.6 WB
GMCV
mg/kg dw
<16.2M
8.15 WB
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Species
Lepomis macrochims
bluegill
Lepomis macrochims
bluegill
Reference
Bryson et al.
1985b
Cleveland et al.
1993
Exposure route and
duration
dietary (lab)
60 days
dietary (lab)
90 days
Selenium form
seleno-DL-cysteine
seleno-L-methionine
lexicological
endpoint
NOEC for juvenile
growth
NOEC for juvenile
survival
Chronic value,
mg/kg dwa
>3.74WBb
>13.4WBb
SMCV
mg/kg dw
GMCV
mg/kg dw
All chronic values reported in this table are based on the measured concentration of selenium in whole body (WB), muscle (M) or liver (L)
tissues.
Chronic value not used in SMCV calculation (see text).
Tissue value converted from ww to dw. See Appendix C for conversion.
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COMPARISON OF FISH CHRONIC CRITERION ELEMENT TO THE MOST
SENSITIVE NON-REPRODUCTIVE SMCV
The egg/ovary chronic criterion element can be compared to the most sensitive non-reproductive SMCV,
cold-stressed (4°C) bluegill, using a conversion from selenium in egg/ovary to whole body tissue. For
bluegill the EO/WB ratio of 2.13 is based on n=27 observations compiled from four different sources
(Coyle et al. 1993; Doroshov et al. 1992; Hermanutz et al. 1996; and Osmundson et al. 2007) and
discussed in Section 4.1.2. These yielded a good correlation between the concentrations of selenium in
egg/ovary relative to whole body. Using an egg-ovary to whole-body ratio of 2.13, a concentration of 15.8
mg/kg dw in the egg-ovary of bluegills converts to a whole-body selenium concentration of 7.4 mg/kg
dw, which is below the 4°C non-reproductive SMCV for bluegill of 8.15 mg/kg dw. Equivalently, the
same whole-body to ovary-egg conversion factor of 2.13 can be used to convert the non-reproductive
bluegill SMCV of 8.15 mg/kg Se whole-body to an ovary/egg concentration of 17.36 mg Se/kg dw, which
would be protected by the reproductive egg/ovary criterion of 15.8 mg/kg.
The bluegill cold-stressed non-reproductive SMCV is also protected by the reproductive effect-based
whole-body criterion of 8.0 mg Se/kg. Figure 16 shows the non-reproductive effect whole-body GMCVs
compared to the whole-body criterion. Figure 17 shows the separate distributions offish nonreproductive
whole-body GMCVs (from Figure 16) and fish reproductive whole-body GMCVs (from Figure 6) in a
single graph.
Being a laboratory investigation of acceptable quality, the Lemly (1993a) 4°C chronic value of 5.85
mg/kg dw was included in the determination of the 8.15 mg/kg dw whole-body non-reproductive SMCV
for bluegill. Nevertheless, the mortality observed in that laboratory study does not appear to be consistent
with field observations. The occurrence of mortality in the field at the concentrations Lemly (1993a)
reported to cause mortality in his lab was not observed in the Lemly (1993b) field study of centrarchid
deformities in Belews Lake. In a field study, Lemly (1993b) found larval centrarchid deformities at
concentrations ranging from 12-80 mg Se/kg dw WB. If juvenile mortality occurred at concentrations
lower than those found to induce larval deformities and at concentrations as low as Lemly (1993a)
reported in the lab (EC40 = 7.91 mg Se/kg WB), then centrarchids would likely not have been present in
Belews Lake. The observations of Lemly (1993b) are evidence that larval deformity, not juvenile
mortality, is the more sensitive endpoint.
The Crutchfield and Person (2000) predictions and field observations of recovery of bluegill at Hyco
Reservoir likewise suggest that significant mortality was unlikely to be occurring at the concentrations
Lemly (1993a) reported to cause substantial mortality. During a time period over which Crutchfield
(2000) indicated dietary invertebrate concentrations exceeded 20 mg Se/kg dw, Crutchfield and Person
(2000) indicated that bluegill population growth occurred at rates predicted to be natural for the
unimpaired species. In contrast, if the Lemly (1993a) lab EC40 of 7.91 mg Se/kg dw whole-body were
applicable to this field situation, the mortality associated with the resulting bluegill whole-body
concentrations (25 mg Se/kg dw whole-body, assuming a trophic transfer factor of 1.27) would have
prevented any recovery.
Selenium-induced cold temperature loss of lipid and body condition, a non-reproductive sublethal effect
that Lemly (1993a) observed to accompany juvenile mortality in the laboratory (but which Mclntyre et al.
(2008) did not observe in a similar study) has also not generally been corroborated by field evidence (Janz
2008). Several studies have measured growth and energy storage indicators in juvenile fish just prior to
and just after winter at reference sites and sites with elevated selenium in northern Canada (Bennett and
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Janz 2007a, b; Kelly and Janz 2008; Driedger et al 2009; Weber et al. 2008). The growth (length, weight,
condition factor, muscle RNA:DNA ratio, muscle protein) and energy storage (whole body lipids, whole
body triglycerides, liver triglycerides, liver glycogen) indicators for five fish species (northern pike,
burbot, fathead minnow, creek chub, white sucker) measured just after winter were similar or greater than
those measured just before winter at the selenium exposed sites. The slimy sculpin did show a decrease in
whole body triglycerides, but the reduction was similar at exposed and reference sites.
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APPENDIX E: OTHER DATA
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SELENITE
Additional data on the lethal and sublethal effects of selenium on aquatic species are presented in Table
E-l. Bringmann and Kuhn (1959a,b, 1976, 1977a, 1979, 1980b, 1981), Jakubczak et al. (1981), and
Patrick et al. (1975) reported the concentrations of selenite that caused incipient inhibition (defined
variously, such as the concentration resulting in a 3% reduction in growth) for algae, bacteria, and
protozoans (Table E-l). Although incipient inhibition might be statistically significant, its ecological
importance is unknown. Albertano and Pinto (1986) found the growth of three red algal species was
inhibited at selenite concentrations that ranged from 790 to 3,958 ug/L.
SELENATE
Dunbar et al. (1983) exposed fed D. magna to selenate for seven days and obtained an LC50 of 1,870
ug/L. This value is in the range of the 48-hr EC50s in Table E-l.
Watenpaugh and Beitinger (1985a) found that fathead minnows did not avoid 11,200 ug/L selenate
during 30-minute exposures (Table E-l). These authors also reported (1985b) a 24-hr LC50 of 82,000
ug/L for the same species and they found (1985c) that the thermal tolerance of the species was reduced by
22,200 ug/L. Westerman and Birge (1978) exposed channel catfish embryos and newly hatched fry for
8.5 to 9 days to an unspecified concentration of selenate. Albinism was observed in 12.1 to 36.9% of the
fry during the five years of such exposures. Pyron and Beitinger (1989) also investigated fathead
minnows, and after a 24-hr exposure, no effect on reproductive behavior was found at 36,000 ug/L, but
when adults were exposed to 20,000 ug/L selenate for 24-hr, edema was observed for their larvae.
The respiratory rate of the eastern oyster, Crassostrea virginica, was unaffected by exposure to selenate at
400 ug/L for 14 days (Fowler et al. 1981). Embryos of the striped bass were quite tolerant to selenate in
dilute salt water (Klauda 1985a, b). There was a 93% successful hatch of embryos at 200,000 ug/L, but
50% of 72-day-old juveniles died after four days at 87,000 ug/L. Exposure of juvenile fish for up to 65
days to concentrations of selenate between 39 and 1,360 ug/L caused developmental anomalies and
pathological lesions.
Do not distribute, quote or cite E-2 Draft Document
-------
Table E-l. Other Data on Effects of Selenium on Aquatic Organisms
Hardness
(mg/L as
Chemical CaCOjl Duration
Effect
Concentration
Reference
Selenium (IV)
Green alga,
Scenedesmus
quadricauda
Green alga,
Selenastrum
capricornutum
Green alga,
Selenastrum
capricornutum
Green alga,
Selenastrum
capricornutum
Alga,
Chrysochromulina
breviturrita
Red alga,
Cyanidium
caldarium
Red alga,
Cyanidioschyzon
merolae
Red alga,
Galdieria
sulphuraria
Algae (diatoms),
Mixed population
Bacterium,
Escherichia coli
Bacterium,
Pseudomonus putida
Protozoan,
Entosiphon sulcatum
Protozoan,
Microreqma
heterostoma
Sodium
selenite
Sodium
selenite
Sodium
selenite
Sodium
selenite
Selenious
acid
Selenious
acid
Selenious
acid
Selenious
acid
Sodium
selenite
Sodium
selenite
Sodium
selenite
Sodium
selenite
Sodium
selenite
FRESHWATER
96hr
72hr
72 hr
72 hr
30 days
20 days
20 days
20 days
18 days
-
16hr
72 hr
28 hr
SPECIES
Incipient
inhibition (river
water)
Decreased dry
weight and
chlorophyll a
BCF= 12-2 lb
BCF= 11,164C
Increased growth
Inhibited growth
Inhibited growth
Inhibited growth
Inhibited growth
Incipient
inhibition
Incipient
inhibition
Incipient
inhibition
Incipient
inhibition
a
2,500 Bringmann and
Kuhn 1959a,b
75 Foe and Knight,
Manuscript
10-100 Foe and Knight,
Manuscript
150 Foe and Knight,
Manuscript
320 Wehr and Brown
1985
3,958 Albertano and
Pinto 1986
3,140 Albertano and
Pinto 1986
790 Albertano and
Pinto 1986
11,000 Patrick etal.
1975
90,000 Bringmann and
Kuhn 1959a
1 1 ,400 Bringmann and
(11,200) Kuhn 1976;
1977a; 1979;
1980b
1.8 Bringmann 1978;
(1.9) Bringmann and
Kuhn 1979;
1980b; 1981
183,000 Bringmann and
Kuhn 1959b
Do not distribute, quote or cite
E-3
Draft Document
-------
Species
Hardness
(mg/L as
Chemical CaCO,) Duration
Effect
Concentration
Reference
Protozoan,
Chilomonas
paramecium
Protozoan,
Uronema parduezi
Snail,
Lymnaea stagnalis
Cladoceran,
Daphnia magna
Cladoceran,
Daphnia magna
Cladoceran,
Daphnia magna
Cladoceran,
Daphnia magna
Cladoceran (<24 hr),
Daphnia magna
Cladoceran
(5th instar),
Daphnia magna
Cladoceran,
Daphnia magna
Cladoceran
(preadult),
Daphnia pulex
Ostracod,
Cyclocypris sp.
Amphipod,
Hyalella azteca
Amphipod
(2 mm length),
Hyalella azteca
Amphipod
(2 mm length),
Hyalella azteca
Amphipod
(2 mm length),
Hyalella azteca
Sodium
selenite
Sodium
selenite
Sodium
selenite
Sodium
selenite
Sodium 214
selenite
Sodium 214
selenite
Sodium 329
selenite
Sodium
selenite
Sodium
selenite
Selenious 220d
acid
Sodium 42
selenite
Sodium 100.8
selenite
Sodium 329
selenite
Sodium 133
selenite
Sodium 133
selenite
Sodium 133
selenite
48 hr
20 hr
7.5 days
48hr
24 hr
24 hr
48 lu-
ge hr
14 days
48 hr
21 days
48hr
48hr
24 hr
48 hr
14 days
48 hr
10 days
24 days
Incipient
inhibition
Incipient
inhibition
LT50
EC50 (river
water)
LC50
EC50
(swimming)
EC50 (fed)
EC50 (fed)
LC50 (fed)
LC50 (fed)
Did not reduce
oxygen
consumption or
filtering rate
LC50
LC50 (fed)
LC50
LC50
(fed)
LOEC
reproduction
(static-renewal)
a
62
118
3,000
2,500
16,000
9.9
710
430
430
685
160
680
1,200
>498
130,000
70
623
312
200
Bringmann and
Kuhn 1981;
Bringmann et al.
1980
Bringmann and
Kuhn 1980a;
1981
Van Puymbroeck
etal. 1982
Bringmann and
Kuhn 1959a,b
Bringmann and
Kuhn 1977a
Bringmann and
Kuhn 1977b
Halter etal. 1980
Adams and
Heidolphl985
Johnston 1987
Kimball,
Manuscript
Reading and
Buikemal980
Owsley 1984
Halter etal. 1980
Brasher and Ogle
1993
Brasher and Ogle
1993
Brasher and Ogle
1993
Midge (first instar),
Sodium
134
48 h
LC50
7,950
Ingersoll et al.
Do not distribute, quote or cite
E-4
Draft Document
-------
Species
Chironomus riparius
Midge (first instar),
Chironomus riparius
Coho salmon (fry),
Oncorhynchus
kisutch
Rainbow trout (fry),
Oncorhynchus
mykiss
Rainbow trout (fry),
Oncorhynchus
mykiss
Rainbow trout,
Oncorhynchus
mykiss
Rainbow trout,
Oncorhynchus
mykiss
Rainbow trout,
Oncorhynchus
mykiss
Rainbow trout
(juvenile),
Oncorhynchus
mykiss
Rainbow trout
(juvenile),
Oncorhynchus
mykiss
Rainbow trout
(juvenile),
Oncorhynchus
mykiss
Rainbow trout
(juvenile),
Oncorhynchus
mykiss
Rainbow trout
(juvenile),
Oncorhynchus
mykiss
Rainbow trout,
Oncorhynchus
mykiss
Rainbow trout,
Oncorhynchus
Hardness
(mg/L as
Chemical CaCO,)
selenite
Sodium 40^8
selenite
Sodium 325
selenite
Sodium 334
selenite
Sodium 334
selenite
Sodium 330
selenite
Sodium 325
selenite
Sodium 325
selenite
Sodium
selenite
Sodium
selenite
Sodium
selenite
Sodium
selenite
Sodium
selenite
Sodium 135
selenite
Sodium 135
selenite
Duration
48 h
43 days
21 days
21 days
5 days
48 days
96 days
4wk
4wk
4wk
42 wk
42 wk
9 days
96 hr
9 days
Effect
LC50
LC50
LC50
Reduced growth
LC50
LC50
LC50
MATC
survival
MATC
survival
BCF = 23
MATC growth
(dietary only
exposure)
MATC survival
(dietary only
exposure)
LC50
LC50
(fed)
Concentration
a
14,600
160
460
250
2,700
2,750
500
280
200
4.7
Hg/g dw
(whole-body)
100
>9.96
Hg Se/g dw
(food)
5.34
Hg Se/g dw
(food)
7,020
7,200
5,410
Reference
1990
Ingersoll et al.
1990
Adams 1976
Adams 1976
Adams 1976
Adams 1976
Adams 1976
Adams 1976
Gissel-Nielsen
and Gissel-
Nielsen 1978
Gissel-Nielsen
and Gissel-
Nielsen 1978
Gissel-Nielsen
and Gissel-
Nielsen 1978
Goettl and
Daviesl978
Goettl and
Daviesl978
Hodson et al.
1980
Hodson et al.
1980
Do not distribute, quote or cite
E-5
Draft Document
-------
Hardness
(mg/L as
Chemical CaCO,) Duration
Effect
Concentration Reference
mykiss
Rainbow trout,
Oncorhynchus
mykiss
Rainbow trout,
Oncorhynchus
mykiss
Rainbow trout,
Oncorhynchus
mykiss
Rainbow trout
(fertilized egg),
Oncorhynchus
mykiss
Rainbow trout
(embryo),
Oncorhynchus
mykiss
Rainbow trout,
Oncorhynchus
mykiss
Rainbow trout
(sac fry),
Oncorhynchus
mykiss
Rainbow trout
(sac fry),
Oncorhynchus
mykiss
Rainbow trout
(egg ),
Oncorhynchus
mykiss
Rainbow trout
(embryo),
Oncorhynchus
mykiss
Rainbow trout
(sac-fry),
Oncorhynchus
mykiss
Rainbow trout
(swim-up fry)
Oncorhynchus
mykiss
Northern pike,
Esox lucius
Sodium
selenite
Sodium
selenite
Sodium
selenite
Sodium
selenite
Sodium
selenite
Sodium
selenite
Sodium
selenite
Sodium
selenite
Sodium
selenite
Sodium
selenite
Sodium
selenite
Sodium
selenite
Sodium
selenite
135 96 hr
9 days
135 41 days
135 50 wk
135 44 wk
120 hr
90 days
272 90 days
272 90 days
96 hr
96 hr
96 hi
96 hi
10.2 76 hr
LC50
(fed)
LOAEC
(Reduced hatch
of eyed embryos)
Decreased iron in
blood and red
cell volume
BCF = 33.2
BCF = 21.1
Did not reduce
survival or time
to hatch
Chronic value for
survival
LC50
MATC
survival
BCF = 17.5
BCF = 3.5
BCF = 3.1
BCF = 3.0
BCF= 13.1
BCF= 1.6
BCF = 80.3
BCF = 20.2
LC50
a
8,200
6,920
26
53
53
10,000
14
55.2e
31.48
0.4
45.6
0.4
45.6
0.4
45.6
0.4
45.6
11,100
Hodson et al.
1980
Hodson et al.
1980
Hodson et al.
1980
Hodson et al.
1980
Klaverkamp et al.
1983b
Mayer etal. 1986
Hunnetal. 1987
Hunnetal. 1987
Hodson et al.
1986
Hodson et al.
1986
Hodson et al.
1986
Hodson et al.
1986
Klaverkamp et al.
1983a
Do not distribute, quote or cite
E-6
Draft Document
-------
Species
Hardness
(mg/L as
Chemical CaCO,) Duration
Effect
Concentration
Reference
Goldfish,
Carassius auratus
Goldfish,
Carassius auratus
Goldfish,
Carassius auratus
Goldfish,
Carassius auratus
Goldfish,
Carassius auratus
Fathead minnow,
Pimephales promelas
Fathead minnow,
Pimephales promelas
Fathead minnow,
Pimephales promelas
Fathead minnow,
Pimephales promelas
Creek chub,
Semotilus
atromaculatus
Bluegill,
Lepomis
macrochirus
Bluegill,
Lepomis
macrochirus
Bluegill (juvenile),
Lepomis
macrochirus
Bluegill (juvenile),
Lepomis
macrochirus
Largemouth bass
(juvenile),
Micropterus
salmoides
Yellow perch,
Percaflavescens
African clawed frog,
Xenopus laevis
Selenium
dioxide
Sodium
selenite
Sodium
selenite
Selenium
dioxide
Selenium
dioxide
Selenium
dioxide
Sodium
selenite
Sodium
selenite
Selenious
acid
Selenium
dioxide
Sodium
selenite
Selenium
dioxide
Sodium
selenite
Sodium
selenite
Sodium
selenite
Sodium
selenite
Sodium
selenite
157
_
_
-
_
157
329
329
220d
_
318
157
16
25
and
200
25
and
200
10.2
_
14 days
10 days
46 days
7 days
48 hr
9 days
96 hr
14 days
8 days
48 hr
48 days
14 days
323 days
120 days
120 days
10 days
7 days
LC50
Mortality
Gradual anorexia
and mortality
LC50
Conditional
avoidance
LC50
LC50
(fed)
LC50
(fed)
LC50
(fed)
Mortality
LC50
LC50
MATC larval
survival
(dietary only
exposure)
No mortality
No mortality
LC50
LC50
a
6,300
5,000
2,000
12,000
250
2,100
1,000
600
420
312,000
400
12,500
19.75
Hg Se/g dw
(food)
>10
10
4,800
1,520
Cardwell et al.
1976a,b
Ellis 1937; Ellis
etal. 1937
Ellis etal. 1937
Weir and Hine
1970
Weir and Hine
1970
Cardwell et al.
1976a,b
Halter etal. 1980
Halter etal. 1980
Kimball,
Manuscript
Kim etal. 1977
Adams 1976
Cardwell et al.
1976a,b
Woock et al.
1987
Lemly 1982
Lemly 1982
Klaverkamp et al.
1983a,b
Browne and
Dumont 1980
African clawed frog, Sodium
1-7 days Cellular damage
Do not distribute, quote or cite
E-7
Draft Document
-------
Hardness
(mg/L as
Species
Xenopus laevis
Chemical
selenite
CaCO,} Duration
Effect
Concentration
a
Reference
Dumont 1980
Selenium (VD
Alga,
Chrysochromulina
breviturrita
Rotifer,
Brachionus
calyciflorus
Snail,
Lymnaea stagnalis
Cladoceran,
Daphnia magna
Cladoceran
(juvenile),
Daphnia magna
Cladoceran
(5th instar),
Daphnia magna
Cladoceran
(5th instar),
Daphnia magna
Amphipod
(2 mm length),
Hyalella azteca
Amphipod
(2 mm length),
Hyalella azteca
Amphipod
(2 mm length),
Hyalella azteca
Amphipod
(1-11 days old),
Hyalella azteca
Amphipod
(1-11 days old),
Hyalella azteca
Midge (first instar),
Chironomus riparius
Midge (first instar),
Chironomus riparius
Rainbow trout
(embryo, larva),
Oncorhynchus
-
Sodium
selenate
Sodium
selenate
Sodium
selenate
Sodium
selenate
Sodium
selenate
Sodium
selenate
Sodium
selenate
Sodium
selenate
Sodium
selenate
Sodium
selenate
Sodium
selenate
Sodium
selenate
Sodium
selenate
Sodium
selenate
30 days
120 96 hr
6 days
129.5 7 days
48hr
48 hr
90 hr
133 48 hr
133 10 days
133 24 days
18 10 days
(S04=3.4)
124 10 days
(SO4=32)
134 48 h
4(M8 48 h
104 28 days
(92-110)
Increased
growth
EC20 Growth
(dry weight)
LT50
LC50
(fed)
LC50
(fed)
LC50
(fed)
42% of
organisms had
visible changes in
gut morphology
LC50
LC50
(fed)
LOEC
reproduction
(static renewal)
LC50
(fed)
LC50
(fed)
LC50
LC50
EC50 (death and
deformity)
50
42.36
(Hg/g dw)
15,000
1,870
550
750
250
2378
627
>700
43
371
16,200
10,500
5,000
(4,180)
(5,170)
Wehr and Brown
1985
Dobbsetal. 1996
Van Puymbroeck
etal. 1982
Dunbar et al.
1983
Johnston 1987
Johnston 1987
Johnston 1989
Brasher and Ogle
1993
Brasher and Ogle
1993
Brasher and Ogle
1993
Borgmann et al.
2005
Borgmann et al.
2005
Ingersoll et al.
1990
Ingersoll et al.
1990
Birge 1978;
Birge and Black
1977; Birge etal.
Do not distribute, quote or cite
E-8
Draft Document
-------
Species
mykiss
Goldfish
(embryo, larva),
Carrassius auratus
Goldfish,
Carassius auratus
Fathead minnow,
Pimephales promelas
Fathead minnow,
Pimephales promelas
Fathead minnow,
Pimephales promelas
Fathead minnow,
Pimephales promelas
Fathead minnow,
Pimephales promelas
Fathead minnow,
Pimephales promelas
Fathead minnow,
Pimephales promelas
Fathead minnow,
Pimephales promelas
Channel catfish
(embryo, fry),
Ictalurus punctatus
Narrow-mouthed
toad
(embryo, larva),
Gastrophryne
carolinensis
Chemical
Sodium
selenate
Sodium
selenate
Sodium
selenate
Sodium
selenate
-
-
-
Sodium
selenate
Sodium
selenate
Sodium
selenate
Sodium
selenate
Sodium
selenate
Hardness
(mg/L as
CaCO,} Duration
195 7 days
24 hr
337.9 48 days
338 48 days
51 30min
24hr
24 hr
44^9 7 days
160-180 24 hr
160-180 24 hr
90 8.5-9 days
195 7 days
Effect
EC50 (death and
deformity)
BCF=1.42
BCF=1.15
BCF= 1.47
BCF = 0.88
BCF= 1.54
LC50
LC50
No avoidance
LC50
Reduced thermal
tolerance
Chronic value -
growth
Chronic value-
growth
Chronic value-
survival
No effect on
reproductive
behavior
Edema in larvae
produced from
adults exposed to
Selenium VI
Induced albinism
EC50 (death and
deformity)
Concentration
a
8,780
0.45
0.9
1.35
2.25
4.5
2,000
1,100
11,200
82,000
22,200
1,739
561
2,000
36,000
20,000
_
90
Reference
1980
Birge 1978
Sharma and
Davis 1980
Adams 1976
Adams 1976
Watenpaugh and
Beitingerl985a
Watenpaugh and
Beitingerl985b
Watenpaugh and
Beitingerl985c
Norberg-King
1989
Pyron and
Beitingerl989
Pyron and
Beitingerl989
Westerman and
Birge 1978
Birge 1978;
Birge and Black
1977; Birge etal.
1979a
Organo-selenium
Bluegill (juvenile),
Lepomis
macrochirus
Seleno-L-
methionine
16 323 days
MATC larval
survival
(dietary only
exposure)
20.83
jig Se/g dw
(food)
Woock et al.
1987
Do not distribute, quote or cite
E-9
Draft Document
-------
Hardness
(mg/L as
Chemical CaCO,) Duration
Effect
Concentration
Reference
Bluegill (juvenile),
Lepomis
macrochirus
Bluegill
(2 yr and adult),
Lepomis
macrochirus
Bluegill
(2 yr and adult),
Lepomis
macrochirus
Redear sunfish
(adult),
Lepomis microlophus
Phytoplankton,
Mixed population
Cladoceran
(<24 hr),
Daphnia magna
Cladoceran
(<24 hr),
Daphnia magna
Midge (<24-hr),
Chironomus riparius
Bluegill (juvenile),
Lepomis
macrochirus
Bluegill (juvenile),
Lepomis
macrochirus
Seleno-L-
methionine
Selenium
Selenium
Selenium
Selenium
Selenite-
Selenate
mixture
Selenite-
Selenate
mixture
Selenite-
Selenate
mixture
Selenite-
Selenate
mixture
Selenite-
Selenate
mixture
283
138
138
138
283
283
90 days
field
field
field
EC20 survival
(dietary only
exposure)
NOEC
deformities
NOEC
deformities
LOEC Adverse
histopathological
alterations
a
>13.4
ug/g dw
(food)
53.83
ug Se/g dw
(liver)
23.38
ug Se/g dw
(ovaries)
<38.15
ug Se/g dw
Cleveland et al.
1993
Reashetal. 1999
Reashetal. 1999
Sorensenl988
Selenium Mixtures
field
21 days
21 days
30 days
60 days
60 days
Reduced growth
rates
MATC
growth
MATC
productivity
MATC
emergence
NOEC survival
EC20 survival
18
115.2
ugSe/L
21.59 ug/gdw
(whole-body)
503.6
340
4.07
ug/g dw
(whole body)
Riedeletal. 1991
Ingersoll et al.
1990
Ingersoll et al.
1990
Ingersoll et al.
1990
Cleveland et al.
1993
Cleveland et al.
1993
Salinity
Chemical (g/kg) Duration Effect
Concentration
Reference
SALTWATER SPECIES
Anaerobic
bacterium,
Sodium
selenite
Selenium (IV)
110 hr Stimulated growth
79.01 Jones and
Stadtman
Do not distribute, quote or cite
E-10
Draft Document
-------
Species
Methanococcus
vannielli
Bacterium,
Vibriofisheri
Green alga,
Chlorella sp.
Green alga,
Platymonas
subcordiformis
Green alga,
Dunaliella
primolecta
Diatom,
Skeletonema
costatum
Diatom,
Chaetoceros
muelleri
Diatom,
Phaeodactylum
tricornutum
Diatom,
Thallassiosira
aestivalis
Brown alga,
Fucus spiralis
Red alga,
Porphyridium
cruentum
Chemical
Sodium
selenite
Sodium
selenite
Sodium
selenite
Sodium
selenite
Selenium
dioxide
Selenium
dioxide
Selenium
dioxide
Selenium
oxide
Sodium
selenite
Sodium
selenite
Salinity
(g/kg) Duration Effect
5 min 50% decrease in
light output
(Micro tox7)
32 14 days 5-12% increase in
growth
32 14 days 23% increase in
growth
32 20 days Increased growth;
induced glutathione
peroxidase
5 days BCF = 18,000
BCF = 16,000
BCF = 10,000
6 days BCF = 337,000
BCF = 65,000
BCF = 5,000
8 days BCF = 109,000
BCF = 27,000
BCF = 7,000
29-30 72 hr No effect on cell
morphology
60 days 1355% increase in
growth of thalli
32 27 days Increase growth;
induced glutathione
peroxidase
Concentration
rug/L)a
68,420
10-10,000
100-10,000
4,600
0.06
0.79
3.6
0.06
0.79
3.6
0.06
0.79
3.6
78.96
2.605
4,600
Reference
1977
Yuetal. 1997
Wheeler et al.
1982
Wheeler et al.
1982
Gennity et al.
1985a,b
Zhang et al.
1990
Zhang et al.
1990
Zhang et al.
1990
Thomas et al.
1980a
Fries 1982
Gennity et al.
1985a,b
Selenium (VI)
Bacterium, Sodium - 15 min 50% decrease in
Vibriofisheri selenate light output
(Micro tox7)
Green alga, Sodium 32 14 days No effect on rate of
Chlorella sp. selenate cell
Green alga, Sodium 32 4-5 days 100% mortality
Chlorella sp. selenate
Green alga, Sodium 32 14 days No effect on rate of
Dunaliella selenate cell population
primolecta growth
Green alga, Sodium 32 14 days 71% reduction in
3,129,288 Yuetal. 1997
10-1,000
10,000
10-100
Wheeler et al.
1982
Wheeler et al.
1982
Wheeler et al.
1982
1,000
Wheeler et al.
Do not distribute, quote or cite
E-ll
Draft Document
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Species
Dunaliella
primolecta
Green alga,
Dunaliella
primolecta
Green alga,
Platymonas
subcordiformis
Green alga,
Platymonas
subcordiformis
Green alga,
Platymonas
subcordiformis
Green alga,
Platymonas
subcordiformis
Brown alga,
Fucus spiralis
Red alga,
Porphridium
cruentum
Red alga,
Porphyridium
cruentum
Eastern oyster
(adult),
Crassostrea
virginica
Striped bass
(embryo),
Morone saxatilis
Striped bass
(larva),
Morone saxatilis
Striped bass
(juvenile),
Morone saxatilis
Striped bass
(juvenile),
Morone saxatilis
Chemical
selenate
Sodium
selenate
Sodium
selenate
Sodium
selenate
Sodium
selenate
Sodium
selenate
Sodium
selenate
Sodium
selenate
Sodium
selenate
Sodium
selenate
Sodium
selenate
Sodium
selenate
Sodium
selenate
Sodium
selenate
Salinity
(g/kg) Duration
32 4-5 days
32 14 days
32 14 days
32 14 days
32 4-5 days
60 days
32 14 days
32 4-5 days
34 14 days
7.2-7.5 4 days
4.0-5.0 4 days
3.5-5.5 9-65 days
3.5-5.5 45 days
Effect
rate of cell
population growth
100% mortality
No effect on rate of
cell population
growth
16% decrease in
rate of cell
population growth
50% decrease in
rate of cell
population growth
100% mortality
160% increase in
growth rate of thalli
23-35% reduction
in rate of cell
population growth
100% mortality
No significant
effect on respiration
rate of gill tissue
93% successful
hatch and survive
LC50 (control
survival= 77%)
Significant
incidence of
development
anomalies of lower
jaw
Significant
incidence of severe
blood
cytopathology
Concentration
10,000
10
100
1,000
10,000
2.605
10-1,000
10,000
400
200,000
13,020
39-1,360
1,290
Reference
1982
Wheeler et al.
1982
Wheeler et al.
1982
Wheeler et al.
1982
Wheeler et al.
1982
Wheeler et al.
1982
Fries 1982
Wheeler et al.
1982
Wheeler et al.
1982
Fowler et al.
1981
Klauda
1985a,b
Klauda
1985a,b
Klauda
1985a,b
Klauda
1985a,b
a Concentration of selenium, not the chemical. Units are Og selenium/L of water unless noted otherwise.
b Converted from dry weight to wet weight basis (see Guidelines)
0 Growth of algae was inhibited
d From Smith etal. (1976).
e Calculated from the published data using probit analysis and allowing for 8.9% spontaneous mortality.
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OTHER DATA - ENDANGERED SPECIES
Two similar studies were conducted in 1996 and 1997 to determine effects of site water and site food,
both contaminated with selenium, on the endangered species, razorback sucker, Xyrauchen texanus
(Hamilton et al. 2001a,b; published later in a peer-reviewed journal in 2005, see Hamilton et al. 2005
a,b,c). Both studies show marked effects of selenium on survival of razorback sucker larvae exposed to
contaminated food and to a lesser extent, contaminated water. Although the data convincingly
demonstrate effects to larval survival from exposure to contaminated food, interpretation of the results in
the context of chronic criterion derivation is complex because of inconsistencies between: 1) levels of
selenium in the food and larvae relative to larval survival; 2) the time to larval mortality relative to
selenium in the diet and selenium in the larvae; and 3) levels of other inorganic contaminants in food and
water (possible organic contaminants were not measured). A summary of each of these two studies is
presented below.
Evaluation of Contaminant Impacts on Razorback Sucker held in Flooded Bottomland Sites Near Grand
Junction , Colorado - 1996 (Hamilton et al. 2001a; also Hamilton et al. 2005 a,b,c)
This study was initiated with 5-day old razorback sucker larvae spawned from adults (first time spawners)
which were previously held (9 months) in three different locations along the Colorado River that
contained varying levels of selenium: Horsethief (the designated reference site which receives water
pumped directly from the Colorado River near Fruita, CO, and where dissolved selenium concentrations
in water ranged from < 1.6 to 3.9 (ig/L during the period of exposure), Adobe Creek (low level selenium
contamination - dissolved selenium concentrations in water ranged from 1.5 to 11.6 (ig/L; avg. = 3.8
(ig/L), and North Pond (high level selenium contamination - dissolved selenium concentrations in water
ranged from 3.8 to 19.6 (ig/L; avg. = 9.5 (ig/L). The selenium content in eggs from three Horsethief
females ranged from 5.8 to 6.6 mg Se/kg dw, and the selenium content in adult muscle plugs at spawning
was from 3.4 to 5.0 mg Se/kg dw. The selenium content in the eggs from three Adobe Creek females
ranged from 38.0 to 54.5 mg Se/kg dw, and the selenium content in adult muscle plugs at spawning was
from 11.5 to 12.9 mg Se/kg dw. The selenium content in the eggs from three North Pond females ranged
from 34.3 to 37.2 mg Se/kg dw, and the selenium content in adult muscle plugs at spawning was from
14.1 to 17.3 mg Se/kg dw. The selenium content in eggs from one of three hatchery brood stock females
was 7.1 mg Se/kg dw, and the selenium content in muscle plugs of two of three hatchery brood stock
females at spawning ranged from 2.6 to 13.8 mg Se/kg dw. The razorback sucker larvae spawned from
fish hatchery brood stock (older, previously spawned females) and held in Colorado River (Horsethief)
water were used as an additional reference group of test fish.
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The experimental groups were subdivided into those receiving reference water (hatchery water; 24-Road
Fish Hatchery) or site water (Table E-2). They were further subdivided into those receiving a daily ration
of reference food (brine shrimp) or zooplankton (predominantly cladocerans and copepods) collected
from each site where their parents were exposed for the previous 9 months. A total of 60 larvae from each
of the four adult sources (Horsethief, Adobe Creek, North Pond, Brood Stock held in different ponds at
Horsethief) were exposed to each treatment (2 replicates x 3 spawns x 10 fish/beaker). The larvae were
held in beakers containing 800 ml of test water. Fifty percent of the test water was renewed daily.
Table E-2. Treatment conditions during the 30-day larval study
Source of Larvae
Horsethief Adults
Adobe Creek Adults
North Pond Adults
Hatchery raised Adults
Treatments
Reference food: Reference
water
Reference food: Site water
Site food: Reference water
Site food: Site water
Reference food: Reference
water
Reference food: Site water
Site food: Reference water
Site food: Site water
Reference food: Reference
water
Reference food: Site water
Site food: Reference water
Site food: Site water
Reference food: Reference
water
Reference food: Site water
Site food: Reference water
Site food: Site water
Se in
food
(mg/kg dw)
2.7
2.7
5.6
5.6
2.7
2.7
20
20
2.7
2.7
39
39
2.7
2.7
5.6
5.6
Dissolved Se in
water
(Og/L)
<1.6
0.9
<1.6
0.9
<1.6
5.5
<1.6
5.5
<1.6
10.7
<1.6
10.7
<1.6
0.9
<1.6
0.9
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Growth, survival and development were evaluated amongst treatment groups for up to 30 days in the
treatment conditions. Each treatment group was fed once daily after renewal. Test waters were collected
every day from each site as grab samples for the renewal. A small portion of this water was retained at 3-
and 7-day intervals for an analysis of total and dissolved selenium concentrations. At approximately 2-
day intervals, aquatic invertebrates and brine shrimp not used for feeding were sieved from the media for
selenium analysis. The number of live fish was recorded daily. After the 30-day exposure period, the
surviving fish were sacrificed and measured for total length. At this same time, approximately four fish
from each treatment, when available, were collected as a composite sample and analyzed for total
selenium.
After 30 days of exposure in the reference food-reference water treatment, survival of razorback sucker
larvae from brood stock and Horsethief adults (89 and 87 percent, respectively) was slightly higher than
those from Adobe Creek adults (84 percent) and North Pond adults (75 percent). Corresponding selenium
concentrations in larval whole-body tissue were 3.6, 3.3, 7.7 and 9.7 mg Se/kg dw, respectively. Survival
was similar or slightly reduced in larvae from all four sources after 30 days of exposure in the reference
food-site water treatments; corresponding selenium concentrations in larval whole-body tissue were 5.2,
5.1, 12.7 and 15.2 mg Se/kg dw, respectively. In contrast, none of the larvae spawned from parents from
Horsethief, Adobe Creek, or North Pond survived to 30 days when fed zooplankton collected from the
three sites, irrespective of the water type they were exposed to (i.e., reference or site). Only the larvae
from brood stock adults, which were fed zooplankton from the Horsethief site for this treatment, survived,
and even these larvae suffered substantial mortality (40 and 60 percent respectively). The mean selenium
concentrations in whole-body tissue of larvae from brood stock adults after the 30-day exposures were 5.4
mg Se/kg dw (site food-reference water treatment) and 6.9 mg Se/kg dw (site food-site water treatment).
Several inconsistencies were observed that indicate selenium may not be solely responsible for the effect
on larval survival. Larval survival in the Adobe Creek treatment group exposed to reference water (<1.6
(ig/L) and reference food (2.7 mg Se/kg dw ) was 84 percent, similar to survival of larvae from brood
stock (89 percent). The selenium concentration in the larvae from this Adobe Creek treatment group after
30 days was higher (7.7 mg/kg dw) than that of the brood stock fish (5.4 mg Se/kg dw) in the reference
water (<1.6 (ig/L) and site food (5.6 mg Se/kg dw) treatment, which had a 30-day survival of 62 percent.
Also, the time to 50 percent mortality between the site food treatments, where most mortality occurred,
was not related to selenium concentration in the diet or in the larvae.
Do not distribute, quote or cite E-15 Draft Document
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Although the larvae from brood stock held at Horsethief and the larvae from the first-time spawning
adults held at Horsethief that were used for the 9 month exposure received the same site food, no larvae
from the latter group survived the 30 day exposure. Concentrations of selenium in the larvae of these two
treatment groups were essentially the same between days 6 and 12 of the exposure (8.1 to 8.9 mg Se/kg
dw). During this same general time frame (6 to 7 days of exposure), larvae from Adobe Creek and North
Pond adults apparently tolerated up to 32 and 39 mg Se/kg dw in tissue, respectively, without any
increase in mortality when exposed to reference food and reference water. Larvae grown out under
hatchery conditions from adults in the Horsethief and Adobe Creek treatments also did not differ in total
deformities compared to larvae from brood stock. There was also no difference between treatments
(brood stock, Horsethief, Adobe Creek, and North pond) in percent egg viability, percent hatchability,
percent embryos with deformities, and percent mortality of deformed embryos and larvae from a separate
test initiated with eggs in the same study (Hamilton et al. 2005b).
Evaluation of Contaminant Impacts on Razorback Sucker held in Flooded Bottomland Sites Near Grand
Junction , Colorado -1997 (Hamilton et al. 200Ib)
In a similar 30-day larval study conducted by the authors in the following year (1997), razorback sucker
larvae from a single hatchery brood stock female (11 mg Se/kg dw muscle) were subjected to the sixteen
different combined water and dietary exposure conditions described in the earlier (1996) study. The
female parent was held at Horsethief Canyon State Wildlife Area before spawning. The larvae were held
in beakers containing 800 ml of test water as before; fifty percent of the test water was renewed daily.
Specific treatment conditions for the 1997 30-day larval study are listed in Table E-3.
Table E-3. Treatment conditions during the 30-day larval study
Water Treatments
Reference food (brine shrimp):
Reference water (24-Road Hatchery)
Reference food: Site water (Horsethief)
Reference food: Site water (Adobe Creek)
Reference food: Site water (North Pond)
Horsethief food: Reference water
Horsethief food: Site water (Horsethief)
Sein
food
(mg/kg dw)
3.2
6.0
32.4
52.5
3.2
6.0
Sein
water
(Og/L)
<1
1.6
3.4
13.3
<1
1.6
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Water Treatments
Horsethief food: Site water (Adobe Creek)
Horsethief food: Site water (North Pond)
Adobe Creek food: Reference water
Adobe Creek food: Site water (Horsethief)
Adobe Creek food: Site water (Adobe Creek)
Adobe Creek food: Site water (North Pond)
North Pond food: Reference water
North Pond food: Site water (Horsethief)
North Pond food: Site water (Adobe Creek)
North Pond food: Site water (North Pond)
Se in
food
(mg/kg dw)
32.4
52.5
3.2
6.0
32.4
52.5
3.2
6.0
32.4
52.5
Se in
water
(Og/L)
3.4
13.3
<1
1.6
3.4
13.3
<1
1.6
3.4
13.3
After 30 days of exposure in this year=s study, there was also good survival of razorback sucker larvae
fed reference food (brine shrimp) and held in reference water or water from Horsethief (83 and 81
percent, respectively). The survival of these larvae was significantly greater than survival of larvae fed
brine shrimp and held in water from North Pond (52 percent). Corresponding selenium concentrations in
larval whole-body tissue after 10 days were 6.3, 6.7, and 11 mg Se/kg dw, respectively. The average
concentrations of selenium in the water for the three treatments were <1, 1.6, and 13.3 (ig Se/L. After 30
days the mean selenium concentrations in these larvae were 5.2, 5.2, and 16 mg Se/kg dw, respectively.
Survival was markedly reduced (0 to 30 percent survival) in the remaining treatments where larvae were
fed zooplankton from the various sites. Complete mortality was experienced by larvae exposed to
Horsethief food and reference water treatment after 30 days.
Similar to the previous study, several inconsistencies in results suggested that selenium may not have
been solely responsible for the effect on larval survival. The most notable inconsistency was that the
greatest effect on larval survival (percent survival or time to 50 percent mortality) was from exposure to
Horsethief food, the food with the lowest selenium contamination.
The authors of the above two studies (Hamilton et al. 2001a,b) make a strong argument that some of the
inconsistency in response observed in their studies between larvae fed reference and site diets may be
related to the difference in arsenic concentration between the two diets. The arsenic concentration
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measured in the brine shrimp used in the reference diet was 24 mg total As/kg dw (measured in the
second larval study) versus between 6 and 7.5 mg total As/kg dw measured in the zooplankton from the
various sites. In their publication (Hamilton et al. 2005c), the authors cite several studies reporting an
ameliorating effect of arsenic against the toxicity of a variety of forms of selenium in various animals
(Dubois et al. 1940, Hoffman et al. 1992, Klug et al. 1949, Levander 1977, Moxon 1938, Thapar et al.
1969). In terms of the survival of larvae from Horsethief, Adobe Creek and North Pond adults when fed
the reference diet, the authors propose that the arsenic concentrations in the brine shrimp diet may have
resulted in an antagonistic interaction with selenium and reduced adverse effects in larvae. Such
hypothesis is questionable, because their studies included diets spiked with inorganic arsenic salts,
whereas the arsenic in brine shrimp (and other natural diets), is most likely predominantly organic arsenic
(US EPA 2003). Additionally, in a separate but related study by the same authors (Hamilton et al. 2005d),
larval razorback sucker spawned from one female at the Ouray Native Fish Facility were fed zooplankton
from six sites (SI, S3, S4, S5, SR, and NR) adjacent to the Green River, Utah at four different initial ages
(5, 10, 24, and 28 day old larvae) for 20 to 25 days. The selenium concentrations in zooplankton from the
SI reference site ranged from 2.3 to 3.5 mg Se/kg dw (dissolved Se in water <0.6 to <1.1 (ig/L). The
concentrations in zooplankton from sites S3 and S4 were slightly higher (range 2.4 to 6.7 mg Se/kg dw;
water, 0.3-0.8 (ig/L), substantially elevated at S5 (12- 26 mg Se/kg dw; water, 0.6-3.1 (ig/L), and highest
at SR and NR (44-94 mg Se/kg dw; water, 14-107 (ig/L). All larvae in the test initiated when they were 5
days old (study 1) died after 25 days of exposure. Median time to death was shortest in fish fed
zooplankton from the reference site (SI) and longest for SR and NR. Interestingly, the concentration of
arsenic measured in zooplankton collected from SI was 12 mg As/kg dw, half that of the brine shrimp
used in the above study (Hamilton et al. 200 Ib), which did not appear to antagonize the toxicity of the
selenium in the diet in this test. In this and the previous two studies, additional inorganic contaminants
such as vanadium and strontium were elevated in the zooplankton fed to the larval razorback sucker.
Do not distribute, quote or cite E-18 Draft Document
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De Riu, D., L. Jang-Won, Huang, S., Monielloa, G., and Hung, S. 2014. Effect of dietary
selenomethionine on growth performance, tissue burden, and histopathology in green and white sturgeon.
Aquat. Toxicol. 148:65-73.
Test Organisms:
Exposure Route:
Test Duration:
Study Design:
Effects Data:
Chronic Value:
Green sturgeon (Acipenser medirostris)
White sturgeon (Acipenser transmontanus)
Dietary only
Three different concentrations of L-selenomethionine were added to an artificial
diet mixture: nominal concentrations of 0 (control), 50, 100, and 200 mg
SeMet/kg (measured: 2.2 mg/kg Se in control diet (no added Se) and 19.7, 40.1
and 77.7 mg/kg Se in the three treatment diets).
8 weeks
Daily rations of the treatment diets (3% BW/d for first 4 weeks and 2% BW/d for
second 4 weeks) were fed to the juvenile sturgeon (approximately 30 g). Each of
the four dietary treatment consisted of 3 replicate 90 L tanks with 25 juveniles in
each tank. Several endpoints were monitored over the 8 week exposure period
including survival, percent body weight increase (% BWI), and hepatosomatic
index (HSI).
White sturgeon had no mortalities through the highest dietary treatment. Green
sturgeon juveniles had 0%, 7.7% and 23.1% mortality with the three dietary
treatments (see table below). %BWI had a greater response to selenium
concentration in juvenile tissues than HSI (see table below). Of note is the
relatively high concentration of Se in the whole body and muscle tissues of the
juvenile sturgeon in the control treatment (both species). The reason for the
relatively high Se control concentrations was not due to accumulation of Se from
the artificial diet because the concentration of Se remained relatively constant
over the 8 week exposure.
TRAP analysis (threshold sigmoid nonlinear regression) of the green sturgeon
survival data resulted in a whole body ECio value of 28.93 mg/kg dw. ECio
values were lower for % BWI and HSI using TRAP. For % BWI, the whole body
ECio value for green sturgeon was 16.36 mg/kg dw, and for white sturgeon,
23.94 mg/kg dw. For HSI, the whole body ECio value for green sturgeon was
10.86 mg/kg dw (with a very wide 95% confidence interval, 1.842-64.08 mg/kg
dw), and for white sturgeon there were no discernible effects.
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Selenium in Juvenile Sturgeon Tissues and Endpoints Measured at end of Eight Week Exposure
Green Sturgeon
Dietary [Se] mg/kg dw
whole body
[Se] mg/kg muscle [Se] survival
dw mg/kg dw % %BWI
HIS
2.2 (control)
19.7
40.1
77.7
7.1
22.8
27.8
34.3
8.4
31.1
37
36.8
100
100
92.3
76.9
6.6
2.6
0.8
-1
2
1.3
0.8
0.9
White Sturgeon
Dietary [Se] mg/kg dw
whole body
[Se] mg/kg
dw
muscle [Se]
mg/kg dw
survival
%
%BWI
HIS
2.2 (control)
19.7
40.1
77.7
5.6
20.1
31.8
47.1
9.2
27
41.3
57.9
100
100
100
100
4.2
4.2
2.8
1
2.6
3.6
3
2.2
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OTHER DATA- CHRONIC STUDIES WITH FISH SPECIES
Some chronic studies met the requirements of an acceptable chronic test but were excluded from being
included in the data set used for criterion derivation for a variety of reasons. Summaries of these studies
are provided below.
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Vidal, D., S.M. Bay and D. Schlenk. 2005. Effects of dietary selenomethionine on larval rainbow trout
(Oncorhynchus mykiss). Arch. Environ. Contamin. Toxicol.49:71-75.
Test Organism:
Exposure Route:
Test Duration:
Study Design:
Effects Data:
Chronic Value:
Rainbow trout (Oncorhynchus mykiss)
Dietary only
Selenomethionine was added to dry fish food; the measured dietary
concentrations were 4.6, 12 and 18 (ig Se/g dw. The measured selenium in the
control diet was 0.23 (ig Se/g dw.
90 days
Each of the three dietary treatments and control had 5 replicates, each replicate
contained 12 to 16 larval rainbow trout that were 27 days old at initiation. Each
fish was fed an average of 10 mg/d for 30 days; 25 mg/d on days 30-60; and 40
mg/d thereafter. Fish were sampled on days 30, 60 and 90 for length, weight,
selenium, hepatic GSH and thiobarbituric acid-reactive substances (TEARS)
measurements.
The authors reported significant decreases in weight and length after the 90-day
exposure (Table E-4). There were no significant differences in the hepatic lipid
peroxidation and hepatic GSH to GSSH ratios among the treatments. The authors
found significant differences in weight and length in the 4.6 and 12 (ig Se/g dw
dietary treatments, but not the 18 (ig Se/g dw treatment. Based on larval trout
body burden, the authors reported an LOEC of 1.20 (ig/g ww, the concentration
of Se in fish fed the 12 (ig Se/g dw dietary treatment. The Se concentration in
larval rainbow trout associated with the lowest dietary treatment that showed
significant decreases in larval weight and length was 0.58 (ig Se/g ww or 2.06 (ig
Se/g dw based on 71.8% moisture in whole body rainbow trout (NCBP).
The data from this study was not used to calculate a chronic value for selenium
due to several inconsistencies. The significant decreases in length and weight
observed in the two lowest concentrations were not observed in the highest
dietary treatment. The Se concentrations in the larval rainbow trout were
irregular with the 60-day concentrations being considerably higher than the 90-
day concentrations. The authors explain this observation to rapid growth in the
fish causing dilution of the Se body burden. However, the increase in fish weight
from 30 to 60 days was similar to the 60 to 90 day increase and the 60 day Se
concentrations increased from day 30. Also, the Se concentration in the control
fish went from below detection on day 0 to 0.46 (ig/g ww on day 30; to 1.24 (ig/g
ww on day 60; and to 0.31 (ig/g ww on day 90. The 60-day measured Se in the
control fish (1.24 (ig/g ww) was more than twice the concentration of Se in the
fish with lowest concentration showing effects (0.58 (ig/g ww).
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Table E-4. Mean (SD) rainbow trout growth after four SeMet dietary treatments
test day
0
30
60
90
Treatment,
Mg/g dw
control
control
4.6
12
18
control
4.6
12
18
control
4.6
12
18
weight, g
0.37(0.30)
1.33(0.92)
1.25(0.21)
1.33(0.30)
1.31(0.37)
2.96 (0.92)
2.33 (0.63)
2.52(0.38)
2.59 (0.24)
5.17(1.09)
3.45 (0.35)*
3.45 (0.35)*
3.82(0.62)
fork length,
cm
3.14(0.41)
4.66(0.41)
4.84 (0.29)
5.09(0.46)
4.97 (0.50)
6.91 (0.56)
6.69 (0.67)
6.88(0.35)
6.92 (0.24)
7.70(0.33)
6.93 (0.19)*
6.84(0.68)*
7.37 (0.62)
[Se] whole body,
(lg/g WW
ND
0.46 (0.20)
1.05(0.77)
1.81(1.04)
1.60(0.93)
1.24(0.54)
1.70(0.72)
1.83(0.94)
2.62(1.22)
0.31(0.20)
0.58(0.21)
1.20(0.21)*
1.41 (0.27)*
[Se] whole body,
Hg/g dw**
ND
1.63
3.72
6.42
5.67
4.40
6.03
6.49
9.29
1.09
2.06
4.25
5.00
* Significantly different than the control.
** ww converted to dw using 71.8% moisture for whole body rainbow trout (NCBP).
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Pilgrim, N. 2009. Multigenerational Effects of Selenium in Rainbow Trout, Brook Trout, and Cutthroat
Trout. Master's Thesis. University of Lethbridge.
Test Organisms:
Exposure Route:
Test Duration:
Study Design:
Effects Data:
Rainbow trout (Oncorhynchus mykiss), cutthroat trout (Oncorhynchus clarkii)
and brook trout (Salvelinus fontinalis)
Dietary only
Selenomethionine added to trout chow and gelatin. Two dietary treatment levels,
nominal Se concentrations, 15 (low) and 40 (high) mg/kg.
Rainbow trout were fed the experimental diets from August - December 2009,
brook trout July - November 2010, and cutthroat trout December 2010 - April
2011.
Fish were obtained from a fish hatchery brood stock. Mature females and were
fed the experimental diets in 710 L tanks. Spawning was stimulated by injecting
Ovaprim® into the females. Eggs were fertililzed and incubated at the fish
hatchery until the eye spots were visible. A portion of the eyed stage larvae from
each treatment was shipped to the University of Lethbridge Aquatic Research
Facility for the swim-up stage of the experiment conducted in gravel bed flumes.
Endpoints measured included percent survival in the first (spawned eggs to eyed
eggs) and second (eyed eggs to yolk-absorbed fry) stages of development, swim-
up success, and malformations (spinal, craniofacial and finfold deformities and
edema).
Selenium affected larval survival, swim-up success and the percent of
malformations in larvae in one or more of the three species tested (see table
below). Visual inspection of plots of the replicate data in Pilgrim (2009) showed
considerable variation between the endpoints and selenium in eggs. The
distribution of selenium among the tissues was markedly inconsistent with other
studies that have used these species. For example, the amount of selenium in the
eggs was 8 and 18 times greater than the concentration in the respective muscle
tissues in cutthroat and rainbow trout. Median ratios (egg Se:muscle Se)
calculated for rainbow trout (Casey and Siwik 2000; Holm et al. 2005) and
cutthroat trout (Golder 2005; Kennedy et al. 2000; Rudolph et al. 2007) were 1.9
and 1.8, respectively. Due to the considerable variation in the concentration
response of the replicate data and anomalous selenium distribution, these data
were not included in the data set to derive the criterion.
Do not distribute, quote or cite
E-24
Draft Document
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Table E-5. Mean selenium concentrations in the diet and selected tissues and selected
endpoints measured in rainbow trout (RN), brook trout (BK) and cutthroat trout (CT).
Adapted from Table 3.1 in Pilgrim (2009).
Species
RBT
BK
CT
Diet ww
1.47
12.7
35.2
1.47
12.7
35.2
1.47
12.7
35.2
Tissue, mg/kg ww
Muscle
0.21
0.51
0.74
0.23
1.14
3.41
0.31
0.93
2.05
Liver
3.77
6.53
17.21
0.72
7.23
20.4
1.00
6.00
14.4
Egg
1.17
4.30
13.0
0.81
5.01
8.15
2.02
9.80
18.0
Survival, %
Stage 1
82.36
77.86
54.72
86.3
71.37
71.37
61.41
30.65
21.99
Stage 2
61.56
48.64
30.33
82.68
88.72
44.63
61.87
14.75
0
Swim-up success
57.18
73.83
27.45
84
83.42
50.11
55.3
21.71
0.08
Total
malformations,
%
10
9.86
29.63
21.3
23.93
24.23
6.13
48.06
NA
Do not distribute, quote or cite
E-25
Draft Document
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Deng, X. 2005. Early life stages of Sacramento splittail (Pogonichthys macrolepidotus) and selenium
toxicity to splittail embryos, juveniles and adults. Doctoral dissertation, University of California, Davis.
Test Organism:
Exposure Route:
Test duration:
Study Design:
Effects Data:
Sacramento splittail (Pogonichthys macrolepidotus)
Dietary only
Four concentrations of selenium in the fish diet (0.6, 17.3, 33.0, and 70.1 mg/g)
were created by mixing different proportions of selenized and Torula yeast. A
different batch of selenized yeast was used in the adult exposure.
24 weeks
Fourteen adult fishes were placed in each circular tank (92 cm diameter, 33 cm
height) and fed one of the four diets. Each diet was provided to fishes in three
tanks. The twelve tanks were arranged in three rows. Each row had all four
treatment concentrations with randomly assigned positions. Thus, the experiment
had a randomized block design. Adult splittail fishes were obtained from the
Tracy Pump Station (U.S. Bureau of Reclamation, Tracy, CA). After 12 and 24
weeks of exposure, blood samples were collected, the liver, gonad, kidney and
white muscle were dissected, and liver and gonad were weighed to calculate
hepatosomatic and gonadosomatic indices. Stages of ovarian and testicular
development were determined from histological studies.
No mortality occurred throughout the experiment. Fish in control, 17.3, and 33.0
mg/g treatments exhibited normal behavior. Fish exposed to 70.1 mg/g in did not
consume as much food as fishes exposed to lower selenium concentrations, and
displayed abnormal behaviors. Splittail adults were less sensitive to dietary
selenium than juveniles. Relative to control, no changes in body weight, total
length, GSI, and condition factor were observed in fishes exposed to selenium
concentrations in food up to 33 mg/g. In general, tissue concentrations in fishes
exposed to selenium were higher than in the control, but differences in selenium
concentrations among them were often small and not significant (Table E-6).
Percentages of ovaries with atretic follicles increased with higher concentrations
of selenium in their diet: 30% in control, 45.5% in the 17.3 mg Se/g, and 100% in
the 33.0, and 70.1 mg/g treatments. The average concentration of selenium in
ovaries offish exposed to 17.3 mg/g in their diet was 6.5 mg/g. This low effect
level, though, is disputable because of the very low number of ovaries analyzed,
the occurrence of atresia in 30% of ovaries in control, and the lack of significant
differences in concentrations of selenium in ovaries among treatments exposed to
elevated levels of this element.
Table E-6. Mean concentration of selenium in ovaries (SE).*
[Se] in ovary (mg/g dw)
Diet Concentration (mg Se/g)
0.6
4.4
(0.57)
17.3
6.5
(1.0)
33.0
8.3
(0.14)
70.1
8.9
(0.46)
' Values estimated from Figure 4 in Deng (2005) (pg. Ill)
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E-26
Draft Document
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de Rosemond, K. Liber and A. Rosaasen. 2005. Relationship between embryo selenium concentration
and early life stage development in white sucker. Bull. Environ. Contamin. Toxicol. 74: 1134-1142.
Test Organism:
Exposure Route:
Test duration:
Study Design:
Effects Data:
Effect
Concentration:
White Sucker (Catostomus commersoni)
Field collected.
In June, 2002, eggs were collected from 4 females from Island Lake (exposed
site); milt was obtained from 2 males. Island Lake is downstream from Cluff
Lake uranium mine located in northern Saskatchewan. Selenium concentrations
in Island lake range from 1 to 11 (ig/L and in recent years have been typically 4-5
(ig/L. No fish/eggs were collected from a reference site.
Through the end of yolk absorption by the larvae; 33 days post-fertilization.
Individual batches of eggs were fertilized in the field with milt and water-
hardened. Eggs were air transported to the laboratory in Saskatoon for testing.
200 eggs were randomly selected from each clutch and then separated into
groups of 100 which were placed into individual test chambers (n = 8).
On test day 30 (3 days prior to test termination), all fish larvae that exhibited
macroscopic deformities (e.g., kyphosis, lordosis, scoliosis and edema) were
removed, photographed and preserved. At test termination, (day 33), 40 larvae
from each female whites sucker were evaluated for deformities using a
microscope.
Although all four females were collected from the exposed site, selenium
concentrations in eggs were grouped into two low (Fish 2 and 3 in Table E-7)
and two high (Fish 1 and 4 in Table E-7). Larval mortality and developmental
deformities were not related to selenium concentrations in eggs (Table E-7). The
data suggest that embryo/larval effects are not observed at concentrations in eggs
reaching 40.3 mg/kg dw (geometric mean of the two high selenium
concentrations in eggs). However, because a reference condition with low
selenium exposure was not established, it is not appropriate to estimate an effect
concentration for this study. Note: the average percent moisture for the four
clutches of eggs was 92.6%.
NA
Do not distribute, quote or cite
E-27
Draft Document
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Table E-7. Embryo/larval endpoints for eggs from four female white sucker collected from Island
Lake in June 2002.
Measurement
Successfully hatched larvae3
Deformed larvaeb
Dead larvae0
Macroscopic deformities , %
Embryologicald
Developmental6
Microscopic deformities, %
Developmental
Total developmental deformities, %8
[Se] eggs mg/kg wwh
[Se] eggs mg/kg dwh
Fishl
161
21
6
6.8
6.2
7.5
13.7
2.7
33.6
Fish 2
140
25
14
6.4
11.4
5
16.4
0.7
9.4
Fish 3
176
16
6
5.7
3.4
2.5
5.9
0.6
8.4
Fish 4
141
13
4
1.4
7.8
7.5
15.3
3.2
48.3
a Initial number was 200 per fish
b Total number of deformed larvae throughout study; includes embryological and macroscopic
deformities
0 Total number of larvae that died throughout study.
d Percent of curled deformities that appeared in embryonic fish; deformities were evident immediately
after embryos hatched.
e Percent of deformities that were designated developmental; deformities became evident as larvae grew
and absorbed yolk sac (after experimental day 15).
f Percent of microscopic developmental deformities that were evident in the 40 fish examined per
female white sucker.
8 The estimated percentage of offspring that had microscopic and macroscopic developmental
deformities combined.
h Selenium concentration measured in a subsample of embryos collected on test day 0.
Do not distribute, quote or cite
E-28
Draft Document
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Ogle, R.S. and A.W. Knight. 1989. Effects of elevated foodborne selenium on growth and reproduction
of the fathead minnow (Pimephales promelas). Arch. Environ. Contam. Toxicol. 18:795-803.
Test Organism:
Exposure Route:
Test Treatments:
Test Duration:
Study Design:
Effects Data:
Fathead minnows (Pimephales promelas; juvenile, 59 to 61 d old)
Dietary only
Purified diet mix spiked with inorganic and organic selenium: 25 percent
selenate, 50 percent selenite, and 25 percent seleno-L-methionine, homogenized
in dextrin.
Completely randomized block design (2 blocks); 4 replicates per block (n = 8
replicates total per treatment). Actual mean total selenium levels in each
exposure treatment were: 0.4 (control), 5.2, 10.2, 15.2, 20.3, and 29.5 mg/kg dw.
Fish used in the first randomized block (F2 generation fish) were progeny from Fj
generation originally obtained from the Columbia National Fishery Research
Laboratory, some of which were used in an initial range-finding experiment. Fish
obtained from a commercial supplier were used in the second randomized block.
The prepared diet was extruded into 1.5 mm pellets which were air-blown dried
to 5 percent moisture content and crushed and sieved so that only particles
retained by an 1 1.8 mesh/cm sieve were used in the study. The amount of
selenium in water that leached from the food during the experiment averaged
only 0.8
105 days, F2 generation (block one) and commercial fish (block two);
14 days F3 generation
Ten fish were randomly placed in each cell per block (n = 8x10, or 80 fish total
per treatment). Fish were fed twice daily at 6 percent body weight per day, with
wastes and uneaten food removed 30 min. after each feeding. Test tanks were
flushed with two tank volumes of fresh test water after each feeding (solution
renewal). Growth (as wet weight) was determined every two weeks by bulk
weighing, and one fish from two of the cells per treatment in a given block (n = 4
total per treatment) was removed for selenium (whole-body) analysis. After 105
days of exposure, a single male and female fish from each treatment replicate (n
= 4 breeding pairs per treatment in a given block, or 8 breeding pairs per
treatment total) were placed in 250 ml beakers and inspected for spawning
activity for 30 days following the first spawning event for that pair (each pair
being one replicate). Gonads and muscle tissue were dissected for selenium
analysis from these fish at the end of the 30 days spawning period. The spawning
substrates were inspected daily for eggs to determine fertility and viability.
Samples of not more than 50 eggs from each spawn were incubated in flowing,
aerated water and inspected for percent hatch determination. Ten larvae from
each incubated brood were transferred to separate glass test chambers and
maintained (48 h renewal; fed brine shrimp twice daily) for 14 days to determine
percent larval survival.
There was no effect of selenium on any of the reproductive parameters measured
at the dietary concentrations tested. Percent hatch and percent larval survival
were very high (>87.4 percent) and essentially equal for all of the treatments.
Growth of pre -spawning adults was affected by the selenium exposure (Table E-
8).
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E-29
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Table E-8. Effects on Fathead Minnow Growth after 98 days of Exposure to Dietary Selenium
Measured mean selenium in
diet, mg/kg dw
0.4
5.2
10.2
15.2
20.3
29.5
Whole-body selenium,
mg/kg dw
1.76
2.78
3.42
5.40
6.58
7.46
Mean fish weight,
g ww
1.30
1.24
1.20
1.21
1.09
0.94
Chronic Value:
An EC value could not be calculated for these data because the data did not meet
the minimum requirements for analysis.
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E-30
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GEI Consultants. 2008. Maternal Transfer of Selenium in Fathead Minnows, with Modeling of Ovary
Tissue to Whole Body Concentrations.
Test Organism:
Exposure Route:
Test duration:
Study Design:
Effects Data:
Fathead Minnow (Pimephales promelas)
Field collected.
Gravid adult fathead minnows were collected from creeks with a wide range of
surface water selenium concentrations near the city of Denver, CO during the
2006 summer breeding season.
Sites
Low selenium exposure:
• Sand Creek at Colfax. In 2002, aqueous selenium averaged 0.9 (ig/L.
Moderate to high selenium exposure:
• Sand Creek downstream of refinery
• East Tollgate Creek
• Mainstem Tollgate Creek
Control fish - no field exposure
• Laboratory-reared fish from Aquatic BioSystems
Embryo-larval test was 48 hours post hatch.
Field collected adult fish were either field dissected for selenium measurement in
paired tissues or transported live back to the laboratory in coolers with site water.
Fish were transported to the laboratory where mating pairs were bred in
individual chambers containing spawning substrates. Eggs were removed from
the spawning substrate and reared in a standard Falcon dish with lab water. Eggs
were screened under a dissecting microscope for viability. Dead eggs were
removed and numbers recorded on a datasheet. Three separate breeding
experiments were conducted.
Upon hatching, larvae were moved to standard bioassay cups containing lab
water and maintained in the laboratory incubator at 25°C. Larvae were
maintained via static conditions in exposure cups for 48 hours post-hatch without
food to ensure full absorption of the yolk sac before they were fixed in formalin.
Deformity assessment was performed on fixed embryos using a dissection
microscope. Test endpoints consisted of egg production, fertilization success,
mortality, and deformities (includes edema and skeletal, craniofacial and finfold
malformations). The authors used a graduated severity index (GSI) for
deformities in which larvae were scored 0 (normal), 1 (slight), 2 (moderate), and
3 (severe) based on the level of defect.
All fish successfully spawned except those collected from Sand Creek
downstream from the refinery. These fish had visible parasites and were only
used in the ovary-to-whole body selenium analysis. A suite of metal and
metalloids were measured in fish samples from each location. Fish collected from
East Tollgate Creek had higher concentrations of 9 of the 15 metals that were
Do not distribute, quote or cite
E-31
Draft Document
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measured in fish from at least one site. Aluminum and iron showed the highest
difference with an approximate 10-fold increase in the East Tollgate Creek fish.
Only the first brood of each mating pair was used for the analysis because effects
appeared to be muted in subsequent broods. The lower response in the second
brood was thought to be due to clearing of selenium in the oocytes. There was
poor correlation between egg fertilization (R2 = 0.13) and embryo mortality (R2 =
0.18) data with whole body selenium concentrations in the adult fish (see Table
E-9 for summary data; see Table E-10 for individual brood data). Neither the
fraction of embryos surviving nor fertilization rate as a function of the
concentration of selenium in maternal fathead minnows was suitable for
estimating EC values. Although there were low survival and fertilization rates at
some higher selenium concentrations, these responses were quite varied and did
not follow a defined concentration-response relationship (Figure E-l).
Of the 9 broods from fish collected at the three exposed sites only one brood
(from East Tollgate Creek) had deformities greater than 10%. The fathead
minnow females that produced the brood with the greatest number of deformities
and highest GSI also had the second highest concentration of whole body
selenium, 46.4 mg/kg dw (Table E-l 1; Figures E-2 and E-3). Approximately half
of the larvae from this brood exhibited some sort of malformation. Similar to the
embryo parameters, EC values were not able to be estimated for any of the 4
malformation parameters.
The authors used probit analysis and TRAP to determine effect levels for each of
the embryonic and larval endpoints (Table E-l2). Although there is an indication
of effect due to selenium exposure in both the embryonic and larval endpoints,
there is too much variation in the responses observed with the embryos and
insufficient response observed with the larvae to derive a reasonable estimate of
effect levels. Therefore, no effect level was determined for this study.
Effect
Concentration: Unable to determine due to high variability or insufficient response.
Do not distribute, quote or cite E-32 Draft Document
-------
Table E-9. Mean fathead minnow first brood embryo and larval parameters and adult whole-body
(WB) selenium concentrations (dw) for each site (± 1SE); CON = control, SCC = Sand Creek at
Colfax Avenue bridge, TGC = Tollgate Creek, and ETC = East Tollgate Creek.
Parameter
n (number of breeding pairs)
WB Se concentration (mg/kg dw)
Egg fertilization (%)
Embryo mortality (%)
Mean spawn size (# of eggs per spawn)
Total larva evaluated (total # of broods)
Mean brood GSI score
Larval craniofacial defects (%)
Larval skeletal defects (%)
Larval finfold defects (%)
Larval edema (%)
Larval length (mm)
Site
Con
10
2.86±0.18
84.75 ±3.32
22.03 ±3.34
129 ±23
957
4.85 ± 1.22
2.64 ±0.90
4.74 ±0.89
2.19 ±0.78
3.89± 1.01
4.90 ±0.05
SCC
3
9.17 ±0.46
23.99 ±22.45
89.04 ±9.70
318±63
89
8. 88 ±8.88
4. 65 ±4.65
9.30 ±9.30
4.07 ±4.07
5.23 ±5.23
4.97 ±0.12
TGC
3
35.87 ±3.73
63 .42 ±31. 82
46.40 ±26.86
162 ±61
281
14.88 ±4.63
6.26 ±3.63
6.21 ±1.48
5.71 ±3.08
6.26 ±3.63
4.83 ±0.14
ETC
4
44.53 ±2.41
59.6 ±22.26
50.76 ±23.63
317 ± 158
254
21.75 ±9.53
18.48 ± 13.84
19.62 ± 12.11
17.23 ± 14.48
20.32 ± 12.93
4.90 ±0.07
Table E-10. Fathead minnow first brood embryo parameters and adult whole-body (WB)
selenium concentrations (dw) for each site (± 1SE); for site acronyms see Table E-9
Total eggs (total
Maternal WB dead+total Survival
Se Cone dw hatch+not fraction (total
Brood Code Treatment (mg/kg) hatched) dead/total eggs)
Fert. Rate ((Initial Egg
Count - 1st day
mortalities)/Initial Egg
Count)
T-la-1
T-lf-1
T-lf-1
T-2a-l
T-3a-l
T-3b-l
T-3d-l
T-4d-l
T-5d-l
T-6d-l
T-2b-l
T-4a-l
T-6a-l
T-2a-l
T-3a-l
T-4a-l
CON
CON
CON
CON
CON
CON
CON
CON
CON
CON
SCC
SCC
SCC
TGC
TGC
TGC
2.90
3.24
1.94
2.25
2.71
2.64
3.67
3.43
3.33
2.52
9.92
8.35
9.25
32.29
43.33
31.99
19
238
19
135
154
90
76
199
149
183
395
193
340
132
79
262
0.79
0.77
0.63
0.98
0.68
0.90
0.70
0.85
0.73
0.76
0.00
0.03
0.30
0.83
0.00
0.77
0.96
0.88
0.73
0.98
0.72
0.95
0.71
0.91
0.87
0.78
0.00
0.03
0.69
0.91
0.00
1.00
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E-33
Draft Document
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Total eggs (total
Maternal WB dead+total Survival
Se Cone dw hatch+not fraction (total
Brood Code Treatment (mg/kg) hatched) dead/total eggs)
Fert. Rate ((Initial Egg
Count - 1st day
mortalities)/Initial Egg
Count)
T-lf-1
T-3b-l
T-5a-l
ETC
ETC
ETC
39.76
47.47
46.37
141
208
634
0.52
0.88
0.07
0.70
0.92
0.17
Table E-ll. Fathead minnow first brood larval malformations and adult whole-body (WB)
selenium concentrations (dw) for each site (± 1SE); CON = control, SCC = Sand Creek
at Colfax Avenue bridge, TGC = Tollgate Creek, and ETC = East Tollgate Creek.
Brood
Code
T-lf-1
T-2a-l
T-6d-l
T-3b-l
T-3a-l
T-la-1
T-lf-1
T-5d-l
T-4d-l
T-3d-l
T-4a-l
T-6a-l
T-4a-l
T-2a-l
T-lf-1
T-5a-l
T-3b-l
Treatmen
t
CON
CON
CON
CON
CON
CON
CON
CON
CON
CON
SCC
SCC
TGC
TGC
ETC
ETC
ETC
Maternal
WBSe
Cone dw
(mg/kg)
1.94
2.25
2.52
2.64
2.71
2.90
3.24
3.33
3.43
3.67
8.35
9.25
31.99
32.29
39.76
46.37
47.47
Total
Larvae
11
141
117
81
96
14
189
95
164
49
3
86
190
91
65
39
150
Spinal
Incidence
9
3
2
4
1
7
8
4
3
6
0
19
5
8
5
44
11
%larvae
w/o
spinal
deformity
91
97
98
96
99
93
92
96
97
94
100
81
95
92
95
56
89
%larvae
w/o
craniofacial
deformity
100
99
99
98
100
93
98
97
98
92
100
91
97
90
95
54
95
%larvae
w/o
finfold
deformity
100
98
99
99
100
93
98
99
99
94
100
92
97
91
98
54
96
%larvae
w/o edema
100
96
97
98
100
93
94
98
96
90
100
90
97
90
94
54
91
Total
GSI
Score
1
24
16
12
1
10
53
20
28
29
0
71
41
78
20
152
89
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E-34
Draft Document
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Table E-12. Authors calculation and comparison of fathead minnow larval deformity
estimates using probit analysis and TRAP.
Effect
Edema
Pinfold
Skeletal
Craniofacial
All
abnormalities
All
abnormalities
except edema
Endpoint
EC10
EC10
ECio
ECio
EC10
EC10
Probit Results
WB [Se]
mg/kg,
dw (±SE)
39.48 ± 16.21
68.55 ±27.26
27.80 ±9.53
53.86± 18.77
16.98 ±5.38
21.35 ±6.45
TRAP Results
WB [Se] mg/kg,
dw (95% CL)
45.78
(40.95-51.20)
48.31
(39.41 -59.21)
46.08
(41.94-50.62)
47.41
(38.92-57.76)
45.50
(41.10-50.37)
45.69
(41.10-50.79)
Probit Results
Ovary [Se]
mg/kg,
dw (±SE)
52.99 ± 19.99
87.95 ±32.16
38.67 ± 12.32
70.83 ±22.84
24.23 ±7.06
30.32±8.51
TRAP Results
Ovary [Se] mg/kg,
dw (95% CL)
61.43
(55.04-68.55)
64.81
(53.01-79.24)
61.82
(56.36-67.80)
63.56
(52.37-77.16)
61.06
(55.26-67.48)
61.27
(55.23-67.97)
Figure E-l. The fraction total survival of embryos (top left), fraction of embryos successfully
fertilized (right), survival adjusted for fertilization (bottom) versus maternal whole
body selenium concentration. Bottom figure EC10=35.2 mg/kg Se dw WB.
1 -
1 0.8-
"0.6-
o
2 0.4 -
0.2 -
t
* «
»** *
• *
^
) 0.5 1 1.5 ;
Log (maternal whole body [Se] \iglg dw)
>
1.00 -
H 0.80 -
c
1 0.60 -
N
m 0.40 -
0.20 -
"•*
* * » » »
*
3 0.5 1 1.5
Log (maternal whole body [Se] \iglg dw)
I
1.2
> *;
Q
>- -
'
0.8 -
£ '€
a £ 0.6 -
01 ^
<; » 0.4 -
K B
1 S. 0.2 -j
| £ o-l-
Z £. 0
0.5
1.5
Log (maternal whole body Se |jg/g dw)
Do not distribute, quote or cite
E-35
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Figure E-2. Percent 2-day post-hatch larvae without edema (A), finfold deformity (B), craniofacial
deformity (C), and spinal deformity (D) relative to maternal whole body selenium
concentration. EClOs: 61.4 - 64.8 mg/kg dw WB.
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Log (maternal whole body [Se] \iglg dw)
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E-36
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Figure E-3. Percent 2-day post-hatch larvae Graduated Severity Index (GSI) relative to maternal
whole body selenium concentration
score
CO
CD
160 -,
140 -
120 -
100 -
80 -
60 -
40 -
20 -
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E-3 7
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Evaluation of zebrafish (Danio rerio) and native cyprinid sensitivity to selenium
Overview:
Two new studies on zebrafish (Danio rerio), Thomas and Janz (2014), Thomas (2014), and Penglase et al.
(2014), were made available to EPA by David Janz, one of the external peer reviewers. Thomas (2014)
and Thomas and Janz (2014) were the original dissertation and peer reviewed paper, respectively, of the
same body of work. The apparent sensitivity of the zebrafish to selenium relative to other species in the
EPA selenium criteria document was the subject of several public commenters, as well as Dr. Janz in the
comments received by EPA.
EPA calculated an EC10 of 7.004 mg Se/kg egg dw, or approximately 3.5 mg/kg whole body) from the
Thomas (2014) and Thomas and Janz (2014) study. EPA was not able to calculate an EC10 from
Pengalese et al. (2014). The Thomas (2014) and Thomas and Janz (2014) study is summarized in the
following section (Part I). Penglase et al. (2014) is summarized in section 7.1.5 of the main document.
EPA noted that the concentration-response curves for both deformities and survival are anomalously
shallow, yielding EClOs far below that of any other sensitive species. The shallow slope indicates partial
effects across the range of test doses, with some individuals being very sensitive, and others being less
sensitive than other test species. A typical test signature of the nutritionally essential element selenium is
that above a particular concentration there is a precipitous increase in adverse effects, with most test
organisms affected within a narrow dose range. Additional issues discovered during the analysis of
available information in the literature and supplied by the investigator raised questions of test quality that
introduced uncertainty in the results reported. This uncertainty, and the fact that zebrafish may not
represent the sensitivity range for cyprinids native to the US (discussed in Part II), led to the decision to
include this study qualitatively in the effects characterization.
The paucity and relative insensitivity of the available data for cyprinids (fathead minnow EC 10 = < 23.9
mg/kg dw; based on LOEC in ovary) relative to other fish families like centrarchids (sunfish), and
salmonids (trout and salmon) caused additional concern. This led EPA to investigate the field
significance of the zebrafish EC 10 (7.004 mg/kg egg) compared to what we know about cyprinid
occurrence in selenium impacted waters. The available studies with native cyprinids indicate that a
variety of native cyprinid genera (e.g. chubs, shiners, dace) have stable, diverse populations and are
reproducing successfully (based on length frequency data) in selenium impacted waters at whole body
concentrations far exceeding our proposed whole body criterion element of 8.0 mg/kg dw. Taken
together, the available studies (Hamilton et al. (1998), NAMC (2008), Presser (2013), USGS (2012)),
indicate that native cyprinids as a family are not expected to be overtly sensitive to selenium when
compared with other families of freshwater fish. This is important because zebrafish are non-native, and
have only been recently discovered in U.S. waters due to accidental introduction.
EPA believes there is significant uncertainty regarding the actual sensitivity to zebrafish, and therefore
proposes inclusion of the zebrafish studies in the effects characterization section, as well as inclusion of a
comprehensive analysis of the studies as well as the studies on sensitivity of selenium to native cyprinids
(below) in its own technical appendix, and issuing an FRN soliciting additional studies or information on
zebrafish, as well as native cyprinids.
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Part I. Chronic summary of Thomas (2014) and Thomas and Janz (2014)
Thomas, J.K.. 2014. Effects of Dietary and in ovo Selenomethionine Exposue in Zebrafish (Danio
rerio). Dissertation. University of Saskatchewan, Saskatoon, Canada.
Thomas and Janz, D.M. 2014. In ovo exposure to selenomethionine via maternal transfer increases
developmental toxicities and impairs swim performance in Fl generation zebrafish (Danio rerio).
Aquatic Toxicol. 152:20-29.
Test Organism:
Exposure Route:
Test Treatments:
Test Duration:
Study Design:
Effects Data:
Zebrafish (Danio rerio)
Dietary only
Selenomethionine spiked into Nutrafin® basic flake food
Control diet (1.3 mg/kg Se dw) and three selenium-spiked diets (3.7, 9.6, and
26.6 mg/kg Se dw).
90 days
Adult zebrafish were fed a control diet (1.3 mg/kg Se dw) and three selenium-
spiked diets (3.7, 9.6, and 26.6 mg/kg Se dw) for 60 days, followed by an
additional 30-40 days with equal rations (2.5%) of control or SeMet-spiked diets
and clean chironomids. After 90 days of feeding exposure, adult fish from each
exposure group were bred 3-4 times and embryos were collected and used to
assess a number of different effects including larval survival and deformities.
Eggs from each treatment were pooled from which replicate samples were
collected for selenium measurement, larval survival and deformity assessment
The authors presented mortality and deformities in the Fl generation graphically
for days up to 6 days post fertilization (dpf). The bar graphics were initially
converted to numeric values using a length measuring tool in GIMP (GNU Image
Manipulation Program). EC 10 values for both mortality and deformities were
very low with deformities being slightly lower. Upon request, the authors
provided a table of the number of deformities in observed in 2-6 days post
fertilization (dpf) fish larvae for each replicate pool of eggs (Table E-13) (David
Janz, pers. comm.). TRAP analysis of these data produced a very low EC10 of
7.0 mg/kg egg Se dw. The concentration-response curve in Figure E-4 is
extremely shallow compared to similar tests on other species, such that the
apparent sensitivity of zebrafish relative to other species depends on what level
of effect is considered. A comparison of egg-ovary zebrafish concentration-
response curves for survival and deformities with well-founded concentration-
response curves for other species is presented in Figure E-5. The shallow
survival and deformity slopes for the zebrafish stand out as atypical for a
selenium response. Note the EC50 values for the zebrafish are very similar to the
EC50 values for the majority of other fish species and the zebrafish EC90 is
similar to the EC90 of the least sensitive fish, Dolly Varden.
A GMCV based on this test has not been included in the Sensitivity Distribution
for several reasons. Although the deformity and survival EC50s are within the
range observed for a number of other species, the concentration-response curves
for both deformities and survival are anomalously shallow, yielding EC 10s far
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E-39
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below that of any other sensitive species (Figure E-5). Furthermore, if the
concentration-response curves are log-symmetrical, as generally has been
assumed in estimating EC 10s, the projected EC90s for zebrafish would place it
among the least sensitive known species. The implication of such a shallow
concentration-response curve is that this species has exceptional genetic diversity
with respect to selenium tolerance, such that populations could adapt to very high
or very low selenium concentrations. The field significance of its exceptionally
low EC 10 is thus uncertain. The low EC 10 might or might not have some
relationship to the selenium deficiency reported by Hook (2008) in substantial
portions of its home range in the Ganges and Brahmaputra basins in India and
Bangladesh.
An assessment of the relative sensitivity of cyprinids using both field and
laboratory data is provided in the following section (Part II).
Table E-13. Selenium concentrations in zebrafish eggs and deformities in 2-6 dpf larvae.
Se in eggs, mg.kg dw
1.67
1.27
1.08
5.99
7.45
6.80
12.26
10.46
15.51
38.98
36.44
26.81
Total
35
63
40
44
45
36
37
39
48
30
65
88
Deformed
0
5
2
6
3
4
11
13
18
21
40
41
% Deformity
0.00
7.94
5.00
13.64
6.67
11.11
29.73
33.33
37.50
70.00
61.54
46.59
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E-40
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1.0
03
V)
O
Q.
03
T3
CD
£S
"03
E
o
0.8 -
0.6 -
0.4 -
o
Q.
o
0.2 -
0.0
0.0
0.5 1.0
log (mg Se/kg egg dw)
1.5
Parameter Summary:
Parameter Initial Final Std. Error
LogXSO 1.45 1.4421 0.0408
Standard Deviation 0.44 0.4421 0.0586
YO 0.95 0.9503 0.0184
Effect Concentration Summary:
%Effect ECx 95%LCL 95%UCL
90 65.15 45.28 93.73
50 27.79 23.08 33.47
20 11.12 8.647 14.29
10 7.004 4.884 10.04
5 5.053 3.208 7.958
95%LCL 95%UCL
1.3632 1.5247
0.3514 0.5964
0.9 0.9799
Figure E-4. Tolerance distribution model (triangular distribution model shape) of the proportion
of normal zebrafish larvae (1-fraction with deformities) vs. the logarithm of concentration of
selenium in zebrafish eggs.
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E-41
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0.5 0.7 0.9 1.1 1.3 1.5 1.7
logfmg Se/kg EO dw)
1.9
2.1
2.3
2.5
Figure E-5. Thomas and Janz (2014) zebrafish concentration-response curves for deformities and
survival, ZF-d and ZF-s, compared with well-founded concentration-response curves for other
species. BG-C: bluegill, Coyle et al. (1993); BG-D: bluegill, Doroshov et al. (1992); BG-H: bluegill,
Hermanutz et al. (1992, 1996); BrnT-su: brown trout survival to swim-up (Formation 2011); CTT-N:
cutthroat trout, Nautilus (2011); DV: Dolly Varden, Golder (2009); LMB: largemouth bass, Carolina
Power & Light (1997); RBT-fc: rainbow trout facial-cranial deformities, and RBT-sk: rainbow trout
skeletal deformities, Holm (2002) and Holm et al. (2003, 2005); Sturg: sturgeon deformities, Linville
(2006).
Part II - Evaluating Sensitivity of Cyprinids (Cyprinidae) to selenium from Field and Laboratory
Data
Background:
The draft selenium criteria document is based on reproductive effects (mortality deformities) to larval fish
following maternal exposure. These chronic tests are based primarily on species from the families
salmonidae and centrarchidae. There is a paucity of data for a number offish families used for
development of selenium criteria. This limitation in data is particularly notable for the family cyprinidae
("minnows"), because it is comprised of approximately 180 general and is one of the most diverse
families in North America. A recent toxicity test with zebrafish (Danio rerio), discussed above in Part 1,
indicated that some cyprinids may be markedly more sensitive to the effects of selenium than other fish
families for which toxicity data are available. This study was very different than all previous studies
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E-42
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examining larval effects in that the slope was very shallow, whereas the slopes for all other species were
steep (see Figure E-5).
This analysis considers the results of the zebrafish laboratory survival study and several field collection
studies, which evaluated cyprinid abundance and diversity in watersheds impacted by selenium, to
compare the sensitivity of the zebrafish evaluated by Thomas (2014) and Thomas and Janz (2014) to
native cyprinid populations. Available water and whole body tissue selenium concentrations (> 8.0 mg/kg
dw), were compared to the translated egg-ovary to whole body zebrafish EC 10 values (~ 3.5 mg/kg dw)
to evaluate the relative sensitivity of native cyprinids to the non-native zebrafish test outcome.
Executive Summary:
The occurrence and effect of selenium on native cyprinids were evaluated based on the results of field
studies conducted in four aquatic systems (CO, NC, UT, and WV) having elevated selenium
concentrations. The objective of this evaluation was to compare the sensitivity of native cyprinid
populations with the results of a recent toxicity test with zebrafish (Danio rerio) (Thomas (2014), Thomas
and Janz (2014)) that suggests some cyprinids may be markedly more sensitive to the effects of selenium
than other fish families for which toxicity data are available. The following set of analyses evaluated
studies of widely-distributed native cyprinid species occurring in waters impacted by selenium from
various sources and the relationships between whole body tissue levels, (and water concentrations where
available) and impacts from selenium via toxicity or population metrics.
Cyprinid genera representing many species native to the US were found to be present in waters with
selenium concentrations exceeding the current national criteria value (5(ig/L). Cyprinid species present in
the four studies examined represent 169 of the approximately 180 species present (at the genus level) in
the United States. Abundance and diversity at sites impacted by selenium (water concentrations > 5.0
(ig/L) were found to be no different than at sites in the Arkansas River, Colorado with low selenium
concentrations (3.0-3.5 (ig/L) watershed, with the exception of one location where extremely high
selenium concentrations (Wildhorse Creek, CO; approximately 413 (ig Se/L) were detected.
Whole body tissue concentrations within several widely distributed cyprinid genera exceeded the
proposed whole body tissue element of 8.0 mg/kg dw and had sustainable reproducing populations, as
indicated by length frequency analysis and occurrence data for the four studies. When evaluated by itself,
the influence of selenium whole-body concentration in reducing family Cyprinidae densities was not
statistically significant (R2 = 0.02; p = 0.51). Rather, substrate characteristics of the waterbodies sampled
had the strongest influence. In contrast, when evaluated by itself, the influence of selenium whole-body
concentration in reducing family Centrarchidae densities was significant (R2 = 0.53; p = 0.02).
In spite of the potential for confounding factors, GEI (2008) obtained parallel results at a different
location, Dixon Creek and Canadian River in Texas, affected by refiner effluent selenium. Again,
selenium whole-body selenium had no relationship to cyprinid density (R2 = 0.00) but was a significant
negative factor for centrarchid density (R2 = 0.41, p = 0.003). And in the Sand Creek Drainage, CO, GEI
found no negative association between fathead minnow densities and selenium concentrations of 3-26 mg
Se/kg whole-body dw and 8-45 mg Se/kg ovary dw.
These findings suggest that native cyprinids are less sensitive than centrarchids, and are thus likely to be
protected by a national criterion based heavily on centrarchid and salmonid sensitivity. Based on these
available data, native cyprinids appear to have a tolerance to selenium that is greater than centrarchid and
salmonid species, and much greater than indicated by the non-native zebrafish test outcome. It is therefore
expected that the proposed selenium criterion will be protective of native cyprinids occurring throughout
the United States.
Do not distribute, quote or cite E-43 Draft Document
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Laboratory Exposures:
1. Chronic Toxicity and Hazard Assessment of an Inorganic Mixture Simulating Irrigation
Drainwater to Razorback Sucker and Bonytail. Hamilton et al. (2000). USGS CERC Laboratory
Toxic effects from inorganics associated with irrigation activities, and possibly contributing to the decline
of endangered fish in the middle Green River, Utah were investigated. Two 90-day chronic toxicity
studies were conducted with two endangered fish, razorback sucker (Xyrauchen texanus) and bonytail
chub (Gila elegans). Swim-up larvae were exposed in a reconstituted water simulating the middle Green
River. The inorganic mixtures were tested at IX, 2X, 4X, 8X, and 16X the measured environmental
concentrations of the evaluated inorganic constituents (2 ug/L arsenic, 630 ug/L boron, 10 ug/L copper, 5
ug/L molybdenum, 51 ug/L selenate, 8 ug/L selenite, 33 mg/L uranium, 2 ug/L vanadium, and 20 ug/L
zinc).
Bonytail chub survival was 95% or greater at 30, 60, and 90 days except for the 16X treatment (1232
ug/L Se), whereas growth was reduced after 30, 60, and 90 days at the 8X treatment (532 ug/L Se).
Swimming performance of bonytail chub was reduced after 90 days of exposure at the 8X treatment.
Whole-body residues of copper, selenium, and zinc increased in a concentration-response manner, but did
not increase at 90 days of exposure at the 8X treatment for most species tested, and at lower treatment
concentrations for the bonytail chub. Mean whole body selenium residues at the 8X treatment were 23.3,
16.7, and 9.4 mg/kg Se dw at 30, 60 and 90 days respectively. Hamilton et al. (2000) concluded that
adverse effects in bonytail chub were associated with whole-body concentrations of 9.4 to 10.8 mg/kg Se
dw in this study. One key uncertainty is the effect that the combination of toxic elements, in contrast to
selenium alone, had on outcomes measured in this study. However, basing the selenium toxicity
evaluation on exposure to multiple contaminants is expected to provide a more conservative estimate of
effect on the bonytail chub (Gila elegans) than if selenium is tested alone.
Field Collection Studies
2. Selenium Tissue Thresholds: Tissue Selection Criteria, Threshold Development Endpoints, and
Potential to Predict Population or Community Effects in the Field. Part III: Field Application of
Tissue Thresholds: Potential to Predict Population or Community Effects in the Field. NAMC
Report (2008).
Field studies were conducted by GEI in the Arkansas River ,CO mainstem and selected tributaries
between 2005 and 2006 to examine the relationship between selenium concentrations as well as habitat
characteristics in surface waters and cyprinid abundance and diversity in the Arkansas River. The data
collected for the study included:
1) seasonal fish and macroinvertebrate (not shown) sampling to determine species composition and the
relative abundance of aquatic organisms);
2) whole-body fish tissue, composite macroinvertebrate tissue (not shown), and water and sediment (not
presented) sample collection for the evaluation of Se concentrations in these tissues and the evaluation of
bioaccumulation pathways; and
3) physical habitat measurements (not presented), to determine relationships between the occurrence of
biota and their physical environment. Data were collected from fall 2004 to fall 2006 from the Arkansas
River, Fountain and Wildhorse Creeks, and the St. Charles River.
Do not distribute, quote or cite E-44 Draft Document
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Total selenium (dissolved) was measured at 4 sites mainstem and 6 sites on three tributaries of the
Arkansas River watershed near Pueblo Colorado (Table E-14). Multiple site visits (6 to 17) to collect
water for selenium determination were conducted at the 10 sampling stations between 2005 and 2006.
Table E-14. Selenium Water Column Data: Total Selenium (ug/L, dissolved)
Site
AR (Arkansas River)
AR1 (ARM) Mainstem, in
Pueblo below Whitlock WWTP
AR2 (ARE) Mainstem below
Pueblo WW Reclamation
Center and Fountain Creek
AR3 (ARE) Mainstem,
downstream of Pueblo
AR4 (ARN) Mainstem,
downstream of St. Charles
River
Arkansas River Tributaries
WHC (Wildhorse Creek)
FC (Fountain Creek)
FCP (Upstream)
FC4 (Downstream)
SC (St. Charles River)
SCI (Upstream)
SC2 (Mid-Point)
SC5 (Downstream)
Sampling Duration
2005-06
8 months
12 months
10 months
10 months
6 months
12 months
6 months
6 months
6 months
8 months
Sample
Size
15
9
7
8
17
9
12
6
11
13
Mean [Se]
(ug/L)
7.05
10.6
8.72
8.81
418
3.43 (4.9)*
12.1
3.09(4.8)*
11.7
20.3
Standard
Deviation
3.69
4.06
4.0
2.85
115
1.05
4.34
1.37
6.22
13
* Maximum [Se] in FCP and SCI < 5.0 ug/L, current selenium criterion
Summary of Selenium Concentrations in Water:
1. Total selenium concentrations exceeded the EPA chronic selenium standard of 5 ug/L in surface water
samples collected from most locations, with only the upper reaches of the St. Charles River and Fountain
Creek having mean selenium concentrations below the EPA chronic selenium standard.
2. Selenium concentrations in water samples from Wildhorse Creek were more than 20X greater than in
water samples collected from all other sample locations, with a mean selenium concentration of 418 ±
115 ug/L.
3. The minimum concentration measured in water samples from Wildhorse Creek (315 ug/L) was
approximately 7X greater than the maximum selenium concentration measured at other study sites (43.6
ug/L at St. Charles River, SC5).
Selenium in Fish Tissue:
Selenium concentrations in fish tissue (whole body) were measured for three representative cyprinid
species (central stoneroller, sand shiner, red shiner), one catostomid (white sucker), and three centrarchids
(green sunfish, smallmouth bass, and largemouth bass) (Table E-15).
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Table E-15. Mean Fish Tissue Concentrations [Average whole body nig/kg dw estimated by eye
from graphs in NAMC (2008)].
Sample Site
Mean water
[Se] ug/L
Cyprinids
Sand Shiner
Red Shiner
Central
Stoneroller
Centrarchids
Green Sunfish
Largemouth
Bass
Smallmouth
Bass
Catostomids
White sucker
ARM
7.0
ARN
8.8
ARE
10.6
ARE
8.7
WHC
418
FCP
3.43
FC4
12.1
SCI
3.1
SC2
11.7
SC5
20.3
10
8
10-21
23
10-20
25
42
18-47
10-17
12
15-21
25
14
5
45
30
33
11-15
7
14-36
22
20
26
20
12
30
40
8-11
10-24
16-18
14-21
32-33
6-10
24
6-14
47
Summary of selenium in fish tissue:
1. The mean concentrations in all cyprinids across all sites was 21.06 mg/kg dwt; SE = 1.38).
2. For comparison, the mean concentration in all centrarchids across all sites was 19.73 mg/kg dw; SE =
1.32; and the mean concentration in white sucker (catostomids) across all sites was 17.52 mg/kg dw; SE =
1.52.
3. Most mean whole-body Se concentrations were well above the U.S. EPA (2014) proposed chronic tissue
criterion element for whole body of 8.13 mg/kg dry weight.
Comparison to national draft fish tissue criteria:
Given that these are waters known to be impacted by selenium there were only a few fish samples (Tables
E-16, E-17) that were at or below the proposed whole body criteria element of 8.1:
1. The Arkansas River mainstem (mean water [Se] = 7.05 ug/L), had samples from three species that met
the criteria in 2006, central Stoneroller, smallmouth bass and white sucker.
2. In the tributaries to the Arkansas River that were sampled, white sucker in both Fountain Creek (mean
water [Se] = 3.43 ug/L) and St. Charles River met the whole body criteria in 2004 and 2005, whereas the
only cyprinid to meet the proposed whole body criterion was the central Stoneroller in 2005.
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Cyprinid Abundance and Diversity:
Table E-16. Cyprinid Diversity (native spp. present- excludes carp): NAMC 2008 Study.
Site
Arkansas River Mainstem
ARM
ARE
ARN
ARE
Arkansas River Tributaries
Fountain Creek
FCP
FC4
Whitehorse Creek (WHC)
St. Charles River
SCI
SC21
SC5
[Se] in water ug/L
7.05
8.72
8.81
10.6
3.43
12.1
413
3.09
11.7
20.3
2005
1/6
6/6
5/6
5/6
5/6
4/6
1/6
5/6
4/6
6/6
2006
3/6
5/6
3/6
4/6
4/6
6/6
1/6
5/6
NS
5/6
:SC2 only sampled in 2005
Table E-17. Cyprinid Abundance (native spp. present- excludes carp): NAMC 2008 Study
Site
Arkansas River Mainstem
ARM
ARE
ARE
ARN
Arkansas River Tributaries
Fountain Creek
FCP
FC4
Whitehorse Creek (WHC)1
St. Charles River
SCI
SC22
SC5
[Se] in water ug/L
7.05
8.72
8.81
10.6
3.43
12.1
413
3.09
11.7
20.3
2005
8
643
697
446
746
1978
926
2920
2757
3102
2006
460
950
521
116
2352
1825
81
14583
NS
2568
'Whitehorse Creek comprised 1 species, central stoneroller
2 SC2 not sampled in 2006
Summary of cyprinid abundance and diversity:
1. Diversity as well as abundance of cyprinids in the tributaries vs the Arkansas River mainstem more likely
a function of habitat and/or predator density rather than influence of selenium.
2. Several sites on Wildhorse Creek, Fountain Creek, and the St. Charles River, had substantial changes in
the populations of some fish species between sample years 2005 and 2006, with fish that were present in
one year in high numbers and with a variety of age classes, either absent or present in low numbers the
other year. These changes are likely to be linked to higher stream flows present in 2006 and significant
habitat changes due to beaver activity at some sites. Variable population compositions and numbers of
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cyprinids are not uncommon in plains streams with highly variable flow regimes and habitat conditions
(Schlosser 1987).
3. Based on an evaluation of age class distribution (indicated by length-frequency distribution data), it was
concluded that the following sites had viable and reproducing cyprinid populations (NAMC 2008:
Arkansas River mainstem: The length-frequency data collected for the fish species at these sites
indicates multiple age groups present for most of the species at the sites.
Fountain Creek - Length-frequency analysis of the flathead chubs indicated that the populations
are reproducing, with juvenile and older adult fish present in relatively high numbers at both sites
and years.
St. Charles River - Length-frequency analysis of the fish populations indicated that sites had
reproducing populations of central stonerollers, fathead minnows, and sand shiners, with juvenile
and adult fish collected during both years (GEI 2007a).
Wildhorse Creek - the age class distribution of central stonerollers was similar between years,
indicating a reproducing population that includes both juvenile and adult fish in both years,
despite the extremely high [Se] in water.
Relevance/Surrogacy of Arkansas River Cyprinids to all Cyprinid Species in US
Cyprinids captured from the Arkansas River are representative of cyprinid species occurring throughout
the US. This conclusion is based on the following lines of evidence:
• Six of the seven cyprinid species (central stoneroller, fathead minnow, flathead chub, longnose dace, red
shiner, and sand shiner) captured from the Arkansas River during this investigation are native to the
United States;
• Four of the six cyprinid species found in the Arkansas River basin (central stoneroller, fathead minnow,
sand shiner and red shiner) are widely distributed throughout the United States (see species specific
distribution maps Attachment 1); and,
• Six of the native species present in the Arkansas River Basin are direct surrogates at the genus level for
the 142 native cyprinids in North America (Table E-l 8).
Table E-18. Cyprinid species surrogacy and occurrence in water for native species inhabiting the
Arkansas River and select tributaries.
Species
Campostoma anomalum
Central stoneroller
Pimephales promelas
Fathead minnow
Platygabio gracilis
Flathead chub
Rhynichthys cataractae
Longnoise dace
Cyprinella lutrensis
Red shiner
Notropis stramineus
Sand shiner
Cyprinid group
stonerollers
Blunthead
minnows
Flathead chub
dace
Satinfm shiners
Eastern shiners
# of species
represented
by genus
5 species
4 species
1 species
9 species
32 species
91 species
[Se] in
waterbodies
where species
occurred
3.1-418 ug/L
3.1 -20.3 ug/L
3.1 - 20.3 ug/L
3.1 -20.3 ug/L
3.1 -20.3 ug/L
3.1 -20.3 ug/L
Average tissue
concentration or
range
5-47 mg/kg dw
No tissue
No tissue
No tissue
23-42 mg/kg dw
10-25 mg/kg dw
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Summary cyprinid surrogacy:
Cyprinid species collected from the Arkansas River watershed are representative (at the genus level) of
the 142 cyprinid species native to North America. With the exception of one sample location (Whitehorse
Creek), the abundance and diversity of cyprinid species present and the occurrence of multiple age classes
indicates that cyprinids are successfully surviving and reproducing in the Arkansas River watershed, even
with selenium concentrations exceeding 5ug/L in water and 8 mg/kg bw in whole body fish tissue. North
American species not represented at the genera level comprise 54 species (mostly chubs - 40 species),
many of which are geographically isolated.
3. Observations of cyprinids in NC Reservoirs (Hyco Reservoir and Belews Lake) - (located at end
of NAMC 2008 report).
Crutchfield et al. (2000) evaluated long-term water quality data, selenium chemical concentration data
collected for sediment, invertebrate and fish tissues, and invertebrate and fish population data collected
from the Hyco Reservoir to document the recovery of the aquatic community following the 1990
installation of a dry fly ash pollution abatement system. Since 1973, data have been collected from six
locations in the Hyco Reservoir, with varying fly ash exposure. Gamefish including bluegill sunfish and
largemeouth bass were reproductively extirpated due to high selenium concentrations prior to installation
of the pollution abatement system, the fish community was dominated by green sunfish (Lepomis
cyanellus), eastern mosquitofish (Gambusia holbrooki), gizzard shad (Dorosoma cepedianum), and
satinfin shiner (Cyprinella analostana). Their main observation was that satinfin shiner was a dominant
cyprinid in the Se limited fish community prior to selenium reduction.
Barwick and Harrell (1997) evaluated fish population monitoring and tissue selenium concentration data
to document the recovery offish populations in Belews Lake for the ten years following installation of a
dry fly ash pollution abatement system. Fish diversity and biomass data were collected from 1977 to
1994 (with the exception of 1978-1979 and 1982-1983) at two sites on the lake. In 1980 and 1981,
fathead minnows (Pimephales promelas) dominated the fish community, representing 62 percent and 81
percent of the biomass, respectively (Barwick and Harrell 1997). By 1984, red shiner (Cyprinella
lutrensis), common carp (Cyprinius carpio), and fathead minnows (Pimephales promelas) were the
dominant cyprinids in the selenium limited fish community prior to selenium reduction. The authors
noted that cyprinid abundance started to decrease as green sunfish, a more Se- tolerant sunfish recovered
in 1989-1990, followed by further decreases in 1990-1994, as channel catfish, bluegill, and largemouth
bass populations increased (Barwick and Harrell 1997).
Young et al. (2010), reviewing the studies of Belews Lake, NC, note that during the period of maximal
selenium inputs, egg and ovary concentrations reached 40-159 mg Se/kg dw. Out of as many as 29
resident species prior to contamination, only catfish and the cyprinids common carp and fathead minnows
remained during the period of maximum impact.
4. Presser, T.S., 2013, Selenium in ecosystems within the mountaintop coal mining and valley-fill
region of southern West Virginia—assessment and ecosystem-scale modeling: U.S. Geological
Survey Professional Paper 1803. 86 p. http://dx.doi.org/10.3133/ppl803.
USGS sampled southern West Virginia ecosystems affected by drainage from mountaintop coal mines
and valleys filled with waste rock (valley fills) in the Coal, Gauley, and Lower Guyandotte watersheds
during 2010 and 2011. Sampling data from earlier studies in these watersheds (for example, Upper Mud
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River Reservoir) and other mining-affected watersheds in WV are also are included to assess additional
hydrologic settings and food webs for comparison.
1. Site-specific fish abundance and richness data documented the occurrence of various species of chub,
shiner, dace, minnow, and central stoneroller (Campostoma anomalum) in the sampled watersheds.
2. Model species for streams were limited to creek chub (Semotilus atromaculatus) and central stoneroller.
Creek chub was present at all sites during USGS sampling in 2010-2011. However, both of these species
are considered to have high tolerance for environmental stressors based on results of traditional
comparative fish community assessments. Concentrations of Se in water and whole body tissues of creek
chub, blacknose dace, and stoneroller are shown in Table E-19.
3. The order of abundance for species with greater than 28 individuals was: creek chub, striped shiner,
mottled sculpin, green sunfish, central stoneroller, blacknose dace, bluntnose minnow, and northern hog
sucker. Shiners and darters were prevalent, but bluegill sunfish were absent during the 2010 survey.
Table E-19. Se in Fish Whole Body Tissue Si
(compilations of data from different sources
Stream Segment
Upper Mud River
Upper Mud River
1
Lower Mud River
Upper Mud River
2 (above Upper
Mud River 1)
Berry Branch
Stanley Fork
Lower Kanawha
River Watershed
Little Scary Creek
Connor Run
Upper Kanawha
River Watershed
Jack's Branch
Mining Complex
Bull push fork
Year
2011
2010
2008
2011
2005
2006
2007
2009-
2010
2009-
2010
[Se] in water
Mean (Range)
in ug/L
10.5, 18.2
Not Sampled
7.9
5.2,7
9.8 (4-22) !
Not Sampled
Not Sampled
8.3 (1.7-18)2
6.0 (3.0-7.4)3
unples: Upper Mud River Basin and Tributaries
presented in (Presser et al. 2013).
Creek Chub
Mean (Range) in
mg/kg dw
9.0(6.4-11)
10.3 (9.4-10.9)
10.3 (9.4-15.4)
9(6.4-11)
2.9(
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Stream Segment
w/pond
Bull push fork
downstream
Hughes Fork
Hughes Creek
Big Coal River
Watershed
Beech Creek
Seng Creek
White Oak Creek
Year
2010
2005-
2007
2010-
2011
[Se] in water
Mean (Range)
in ug/L
9.1-10
5.3 (2-10)
2.1-13
Creek Chub
Mean (Range) in
mg/kg dw
8.6(6.2-13)
7.8(4.1-10.9)
2005
7.9 (2.7-12.9)
2007
9.9(3.7-17)
Blacknose
Dace Mean
(Range) in
mg/kg dw
10.7(5.5-14)
Not Sampled
16.9 (6.8-25)
Stoneroller
Mean (Range)
in mg/kg dw
6.9(3.1-17)
12.4 (0.5-34.5)
2005
9.0 (3.6-14)
2005-
2007
2005-
2009
2011
2005-
2007
Not Sampled
27.5 (15-42)
23.3
15.8
(8-27)
(3-18)
8.2(4.8-14.7)
8.1(5.4-10)
5.8(<1-12.8)
Not Sampled
Not Sampled
Not Sampled
Not Sampled
Not Sampled
Not Sampled
Not Sampled
7.1 (2.5-12.8)
1 Water samples collected between 2005 and 2008.
2 Water samples collected in 2009 and 2010.
3 Water samples collected in 2009 and 2010.
Study Summary:
Samples in various environmental media (water, sediment, algae, macroinvertebrates, fish) were collected
by USGS (2010-2011), and others (e.g. WVDEP, Potesta) between 2005 and 2011. The stream segments
presented here represent a subset of the stream segments with available data. Only streams with water
[Se] > 5.0 ug/L are presented to facilitate comparison with other studies with Se-impacted streams.
Overarching observations include:
1. [Se] in water averaged from 5.3 ug/L - 31.4 ug/L with a high of 90 ug/L (Connor Run, 2009).
2. [Se] in fish tissue: creek chub - averaged from 5.8 mg/kg wb to 28 mg/kg wb, with a maximum whole
body concentration of 80 mg/kg wb (Little Scary Creek, 2009).
3. [Se] in fish tissue: blacknose dace - averaged from 10.7 mg/kg wb to 66 mg/kg wb, with a maximum
whole body concentration of 113 mg/kg wb (Bull push fork w/pond, 2010)
4. [Se] in fish tissue: central Stoneroller - averaged from 6.9 mg/kg wb to 12.4 mg/kg wb, with a maximum
whole body concentration of 34.5 mg/kg wb (Hughes Fork, 2005). Note also, that central Stoneroller,
although common through stream segments samples, were not ubiquitous, as was observed in the study
conducted by NAMC in the Arkansas River near Pueblo CO.
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5. Selenium concentrations in fish tissue collected from the Gunnison River
River. http://pubs. usgs.gov/of/2012/1235/ofl2-1235.pdf
Approach: In sampling conducted in summer 2010, muscle tissue plugs were collected from common
carp (Cyprinus Linnaeus), roundtail chub (Gila robusta; listed), and whole body tissue samples were
collected from speckled dace (Rhinichthys osculus) inhabiting critical habitat in the Gunnison River in
Western Colorado. Total selenium in fish muscle plugs (mg/kg dw) for roundtail chub, or in whole body
(speckled dace) was calculated for all tissues. In follow-up sampling conducted in the summer of 2011,
muscle plugs were collected from common carp (Cyprinus Linnaeus), roundtail chub (Gila robusta;
listed), and bonytail chub (Gila elegans, listed) inhabiting critical habitat in the Gunnison River in
Western Colorado.
This study was intended to document any changes in selenium concentration in fish over the last 20 years
based on remediation efforts that have been completed to date.
Table E-20. Fish Tissue Concentrations observed in Cyprinids
Species
Roundtail Chub
Speckled Dace
Year
2010
2011
2010
Mean (Range) [Se]
9. 7 mg/kg dw (5. 2-32.4)
7.33 mg/kg dw (5. 6-1 1.2)
7.46 mg/kg dw (5. 7-9.7)
# > muscle = 11*
2/15
1/15
# > whole body = 8
6/15
* Muscle plugs were collected since this species is aO large enough for non-destructive sampling, and b) a
listed species.
OTHER DATA- CHRONIC STUDIES WITH INVERTEBRATE SPECIES
A limited number of studies have evaluated the effects of selenite on invertebrate species, an important
prey item for fish and birds as summarized by Debruyn and Chapman (2007). The following studies with
a rotifer, and annelid, and an insect (mayfly) were found suitable for establishing species sensitivity.
Dobbs et al. (1996) exposed Brachionus calyciflorus to selenate in natural creek water for 25 days in a
three-trophic level food chain test system. This is one of two laboratory-based experiments (also see
Bennett et al. 1986) that involved exposing algae to selenium (in this case as sodium selenate) in water,
and subsequently feeding the algae to rotifers which were in turn fed to fish (fathead minnows). In this
particular study, the rotifers and fish were exposed to the same concentrations of sodium selenate in the
water as the algae, but received additional selenium from their diet (i.e., the algae fed to rotifers and the
rotifers fed to fish). The overall exposure lasted for 25 days. Rotifers did not grow well at concentrations
exceeding 108.1 (ig Se/L in water, and the population survived only 6 days at selenium concentrations
equal to or greater than 202.4 (ig Se/L in the water (40 (ig Se/g dw in the algae). Regression analysis of
untransformed growth data (dry weight) determined 4 day post-test initiation resulted in a calculated ECi0
of 37.84 (ig Se/g dw tissue.
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Although not intended to be a definitive toxicity study for this invertebrate, Besser et al. (2006) evaluated
the bioaccumulation and toxicity of selenized yeast to the oligochate, Lumbriculus variegatus, which was
intended to be used for dietary exposure in subsequent studies with the endangered desert pupfish,
Cyprinidon macularius. Oligochaetes fed selenized-yeast yeast diets diluted with nutritional yeast (54 to
210 mg Se/kg) had stable or increasing biomass and accumulated Se concentrations as high as 140 mg/kg
dw. The oligochaetes fed the undiluted selenized-yeast (826 ug/g Se dry wt.) showed reduced biomass.
The effect level is considered >140 mg Se/kg dw.
Conley, J.M., D.H. Funk and D.B. Buchwalter. 2009. Selenium bioaccumulation and maternal transfer in
the mayfly Centroptilum triangulifer in a life-cycle, periphyton-biofilm trophic assay. Environ. Sci.
Technol. 43:7952-7957.
Conley, J.M., D.H. Funk, N.J. Cariello and D.B. Buchwalter. 2011. Food rationing affects dietary
selenium bioaccumulation and life cycle performance in the mayfly Centroptilum triangulifer.
Ecotoxicol. 20:1840-1851.
Conley, J.M., D.H. Funk, D.H. Hesterberg, L-C. Hsu, J. Kan, Y-T. Liu and D.B. Buchwalter. 2013.
Bioconcentration and biotransformation of selenite versus selenite exposed to periphyton and subsequent
toxicity to the mayfly Centroptilum triangulifer. Environ. Sci. Technol. 47:7965-7973.
Conley et al. (2009) exposed mayfly larvae (Centroptilum triangulifer) to dietary selenium contained in
natural periphyton biofilms to eclosion. The periphyton fed to the mayfly larvae were exposed to
dissolved selenite (radiolabeled 75Se) in November 2008 (12.6 and 13.9 (ig/L) and in January 2009 (2.4,
2.4, 4.9, 10.3, and 10.7 (ig/L). Periphyton bioconcentrated Se an average of 1113-fold over the different
aqueous Se concentrations (Table E-21). Twenty 4 to 6-day old mayfly larvae were exposed for 4.5 to 6
weeks to each of the periphyton diets until the larvae eclosed to subimagos. The subimagos were allowed
to emerge to the adult imago stage which deposited their egg masses in Petri dishes. Selenium was
measured in postpartum adults along with their dry weights and clutch size.
Table E-21. Selenium Concentrations in Water Exposed to Periphyton, Periphyton and Mayfly
Adults
Treatment
5A
5B
10A
20C
20D
20A
20B
Dissolved [Se] exposed
to periphyton, jig/L
2.4
2.4
4.9
10.3
10.7
12.6
13.9
[Se] in periphyton,
mg/kg dw
2.2
2.0
4.4
8.7
11.3
25.5
17.5
[Se] in mayfly adult,
mg/kg dw
4.2
5.7
9.7
16.2
27.5
56.7
34.8
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Selenium increased in concentration from periphyton to the adult mayflies (trophic transfer factor) an
average of 2.2-fold (Table E-21). The authors observed a decrease in fecundity as maternal postpartum Se
concentrations increased. Fecundity was also related to growth of the mayflies. The authors observed a
reduction in fecundity for this mayfly when they were fed diets containing more than 11 mg Se/kg dw.
This threshold is considered the effect value for this study. Using the trophic transfer factor of 2.2, the
periphyton Se concentration of 11 mg/kg dw translates to an adult mayfly Se concentration of 242 mg/kg
dw.
Conley et al. (2011) exposed larval C. triangulifer similar to Conley et al. (2009) to two different rations
of periphyton (Ix and 2x) to evaluate the effect of feeding ration on the bioaccumulation and life cycle
performance of the mayfly. Periphyton (on plates) was initially exposed to low (1.1 to 3.4 (ig/L), medium
(5.9 - 8.9 (ig/L) and high (19.2 - 23.1 (ig/L) selenite. Fifteen 1-2 day-old mayfly larvae were then fed
either 1 plate (Ix ration) or 2 plates (2x ration) in bottles containing 1.8 L water to eclosion to subimagos
(25-29 days). Subimagos were induced to emerge to adults in petri dishes and their clutch size measured
through digital imaging. Selenium measurements from this study are given in Table E-22.
Table E-22. Selenium concentrations in water, periphyton and mayfly tissues for two feeding
rations (adapted from Table 1 in Conley et al. 2011)
Feeding ration - Se level
Ix - low
Ix - medium
Ix - high
2x - low
2x - medium
2x - high
Mean dissolved Se
exposed to
periphyton, u£/L
1.1
5.9
21.4
2.7/3. 4a
7.1/8.93
19.2/23. la
Mean periphyton, mg
Se/kg dw
4.2 ± 0.6 (4)
11.9 ±2.1 (4)
27.2 ± 4.2 (4)
9.5 ±0.9 (3)
19.9 ±1.6 (3)
40.9 ±1.7 (3)
Mean mayfly tissue,
mg Se/kg dw
12.8 ±3.6 (28)
31.7±7.5(15)
68.4 ±24.0 (9)
14.1 ±3. 8 (19)
21.6 ±2.8 (22)
37.3 ±6.7 (13)
a Two values represent two different loading exposures, September and October. The plates were
combined for mayfly exposure.
Mayflies fed the Ix ration had 54% and 72% reductions in survival relative to controls in the medium and
high Se treatment levels, respectively, both significant (p<0.05). The mayflies fed the Ix ration also had
significant reductions in fecundity in the low (44% reduction), medium (63% reduction) and high (77%
reduction) Se treatment levels. However, for the mayflies fed the 2x ration, there were no significant
differences between the controls and any of the three Se treatment levels for any of the endpoints
measured including survival and fecundity. The 2x ration mayflies had 60% more biomass than the Ix
ration mayflies. This growth difference explains why the Ix ration mayflies had higher concentrations of
Se in their tissues. The two different rations resulted in vastly different effect levels for Se, <12.8 mg/kg
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dw in the Ix ration test and >37.3 mg/kg dw in the 2x ration. It is apparent from this study that if the
mayflies do not obtain sufficient nutrition, they are more sensitive to selenium. Although reduced feeding
levels occur in nature, it is a confounding variable in this study that cannot be used to set a chronic effect
level for selenium.
Conley et al. (2013) evaluated the accumulation of selenite and selenate into periphyton with subsequent
feeding exposure to mayfly larvae. As in his previous studies, C. triangulifer larvae were fed periphyton
previously exposed to different concentrations of selenium. In this study, periphyton plates were first
exposed to low (10 (ig/L) and high (30 (ig/L) concentrations of either selenite or selenate and then fed to
mayfly larvae to ecolsion to subimagos. The selenite and selenate treatment exposures resulted in similar
levels of selenium in the subimagos. Since no differences in selenium accumulation was observed, the
selenite and selenate treatments could be pooled for measuring the endpoints, survival and secondary
production (total mayfly biomass produced). Mean selenium concentrations fed the mayflies were 2.2,
12.8 and 37 mg/kg Se dw in the control, low and high treatments, respectively. Mayfly tissue (subimago)
concentrations (extrapolated from Figure 4a in Conley et al. 2013) were approximately 4-7, 20-35, and
45-75 mg/kg Se dw, in the control, low and high treatments, respectively. The authors reported
significant reductions in survival from the control in the high Se treatment (both pooled data and
individual selenite and selenate treatments) but no significant differences were observed in the low Se
treatments. Secondary production was significantly reduced relative to the control in the high Se
treatment for both selenium species. For the low Se exposure treatment, secondary production was not
significantly different than the control for the selenite treated periphyton exposure, but was for the
selenate and pooled data suggesting an effect level between 20 and 35 mg/kg Se dw. These results as
well as those observed in 2x ration exposures in Conley et al. (2011) where no effects were observed at
37.3 mg/kg Se dw generally support the chronic value determined for Conley et al. (2009) of 24.2 mg/kg
Se dw.
The following invertebrate studies were inconclusive for establishing species sensitivity because of
limitations in the experimental designs, as explained for each.
Malchow et al. (1995) fed fourth instar Chironomus decorus midge larvae a diet of seleniferous algae
under laboratory conditions for 96 hours. For algae cultured with selenite, a larval tissue concentration of
4.05 (ig Se/g dry weight resulted in a 46% reduction in growth relative to the controls. At a larval tissue
concentration of 8.6 (ig Se/g dry weight, larval growth was reduced by only 39%. Since the study only
reported two exposure concentrations, it is unclear if the tissue effect concentration at 4.05 (ig Se/g dry
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weight is real or an anomaly. Additional exposure concentrations and subsequent effect levels are needed
to resolve this issue.
Malchow et al. (1995) also fed fourth instar Chrionomus decorus midge larvae a diet of algae cultured
with selenate, and the midge larvae were exposed under laboratory conditions for 96 hours. A dietary
exposure of 2.11 (ig Se/g dry weight significantly reduced larval growth (15% reduction) at tissue
concentrations of 2.55 (ig Se/g dry weight. At a larval tissue concentration of 6.62 (ig Se/g dry weight,
growth was reduced 20% relative to the controls. The 15-20% reduced growth at larval tissue
concentrations 2.55 (ig Se/g dry weight may be statistically significant, but not biologically meaningful.
In addition, exposure to only two selenium concentrations precludes confirmation of a dose-response.
Alaimo et al. (1994) also exposed 2010 midge larvae to selenite diet, but the selenium source was from
field contaminated widgeongrass (Ruppia maritimd). Ruppia stems and leaves were collected from four
selenium contaminated evaporation ponds located in the San Joaquin Valley of California. Three-day old
larvae were exposed to each of the four treatment diets (Ruppia from each pond) plus a Cerophyll control
for 14 days (egg to pupation), with the moderately hard reconstituted water renewed at day 7 and every
three days thereafter. The growth (weight) of exposed larvae was significantly reduced in all of the
selenium treatments when compared to the controls. The lowest effect level was observed for the
Westlake pond (primarily selenite), where growth was reduced 40 percent relative to the controls at a
larval tissue concentration below the detection level (1.0 ppm dry weight, or 1.0 (ig Se/g dry weight).
These results are suspect because the field collected Ruppia likely contained contaminants other than
selenium, the control organisms were fed a different diet (Cerophyll), and the single concentration
exposure is difficult to defend.
OTHER DATA - FIELD STUDY WEST VIRGINIA IMPOUNDMENTS
In response to the USEPA (2004) draft whole fish tissue criterion for selenium, the West Virginia
Department of Environmental Protection (2010) initiated a study to assess selenium bioaccumulation
among fishes residing in the State's lakes and streams. A focus of the study was the collection and
evaluation of bluegill, Lepomis macrochirus, larvae (ichthyoplankton) from selected waterbodies since
2007, based on concerns regarding fish population health at locations subjected to elevated selenium
inputs, particularly during the more sensitive developmental life stages of fishes (e.g. yolk-sac larvae).
Also, in 2009, WVDEP began acquiring data about selenium concentrations within fish eggs of various
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species within reference and selenium-impacted waters. WVDEP also conducted deformity surveys of
adult fishes in selenium enriched waters as well as at reference locations in 2008-2009.
WVDEP scientists found that larval deformity rates were variable throughout the study duration but were
nonetheless correlated with waterborne selenium exposure. Reference locations produced age-based
larval bluegill subsamples (24-168 hours) with low deformity rates (0 - 1.27%); whereas, locations with
seleniferous inputs exhibited bluegill deformity rates ranging from 0% to 47.56% in developmental stages
up to 312 hours. Maximum deformity rates among staged bluegill subsamples as determined through
these evaluations were 19.28%, representing specimens collected from selenium-enriched waters.
Concentrations of selenium within fish eggs also varied according to study location and ranged from <0.8
mg/kg dry weight among bluegill eggs at the control site to 64.62 mg/kg dry weight among largemouth
bass, Micropterus salmoides, eggs collected from selenium-enriched waters. Searches for more mature,
yet developmentally-deformed fishes revealed increased deformity rates (14%) among largemouth bass
residing in a selenium impacted reservoir as compared to deformity rates among largemouth bass found in
the reference lake (0%). The data on egg selenium concentrations are not adequate for constructing a
concentration-response curve. Nevertheless, the overall deformity rate in the contaminated Upper Mud
River Reservoir was 5% among 10,000 individual fish, average egg selenium concentration 9.8 mg/kg
dw. The overall deformity rate in the reference Plum Orchard Lake was 0.5% among 13,000 individuals,
average egg selenium concentration nondetectable or <0.8 mg/kg dw.
OTHER DATA - NUTRITIONAL DEFICIENCY/SUFFICIENCY STUDIES
CONTAINING MEASURED SELENIUM IN THE DIET AND WHOLE
BODY FISH TISSUE
Ingested dietary dose studies in fish designed to identify nutritionally deficient and/or nutritionally
sufficient selenium doses in fish food or prey primarily describe selenium effects on growth, with survival
reductions and effects on antioxidant enzyme activity also occasionally reported. A number of the dietary
studies have measured a range of dietary doses that maximize fish growth, as opposed to a single dietary
dose associated with nutritional sufficiency for growth. Regardless of whether nutritionally sufficient
dietary doses are reported as a single concentration or as a range of concentrations, reduced growth or
survival is observed at both lower dietary doses (nutritional deficiency) and at higher dietary doses
(toxicity).
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Although dietary doses are normally presented as selenium concentrations in food, expressed in terms of
mg/kg Se in the diet, several studies have also concurrently presented nutritionally deficient/sufficient Se
levels in terms of the whole body Se concentration in the fish. These studies permit a comparison of
nutritionally deficient/sufficient whole body Se residues in fish to the national criterion for Se in whole
bodies offish. When combined with measured whole body fish tissue residues associated with toxicity, a
complete picture of the range of Se residues in whole body fish tissue associated with nutritional
deficiency, nutritional sufficiency and toxicity emerges.
Eight fish species have information on both nutritionally deficient dietary doses and whole body
concentrations of selenium measured in the same study (Table E-23). Six of the eight species are native
to North America. Nutritionally deficient dietary doses of Se range between 0.03 mg/kg dw in Atlantic
salmon (Salmo salar, Poston et al. 1976) associated with reduced survival to 1.4 mg/kg dw in Atlantic
cod (Gadus morhua, Hamre et al. 2008), also associated with reduced survival. Whole body Se residues
identified as nutritionally deficient range between 0.64 mg/kg dw in Malabar grouper (Epinephelus
malabaricus) associated with suboptimal weight gain and feed efficiency (Lin and Shiau 2005) and 4.72
mg/kg dw in North African catfish (Clarias gariepinus), also associated with suboptimal weight gain
(Abdel-Tawwab et al. 2007). The whole body Se residues associated with growth and/or survival
reductions due to nutritional deficiency of the six North American species (Prussian carp, Han et al. 2011;
common carp, Gaber 2007; Atlantic cod, Hamre et al. 2008; Coho salmon, Felton et al. 1990; cobia, Lin
et al. 2010; Atlantic salmon, Poston et al. 1976) all range between 1.0 and 2.7 mg/kg dw.
Ten fish species have information on both nutritionally sufficient dietary doses and whole body
concentrations of selenium measured in the same study (Table D-23). Eight of the 10 species are native
to North America. Nutritionally sufficient dietary doses of Se for the North American resident species, all
but one of which are based on maximum growth offish, range between 0.1 mg/kg dw in hybrid striped
bass (Jaramillo 2006) and 6.6 mg/kg dw in rainbow trout (Hilton and Hodson 1983). Several studies have
identified a range of dietary doses and associated whole body residues that maximize growth and survival
relative to that offish fed lower dietary doses and which subsequently contain lower whole body selenium
residues. Whole body Se residues associated with nutritional sufficiency based on maximal growth
and/or survival of all North American species except for hybrid striped bass (Jaramillo 2006) range
between 0.2 - 3.63 mg/kg dw (Table D-23). For hybrid striped bass, Jaramillo (2006) observed that
maximum weight gain occurred in selenite supplemented diets containing 1.19 mg/kg dw Se, which
resulted in whole body Se residues of 5.13 mg/kg dw. Jaramillo (2006) also exposed hybrid striped bass
to seleno-DL-methionine supplemented diets containing 0.90 mg/kg dw, which resulted in the maximum
Do not distribute, quote or cite E-58 Draft Document
-------
weight gain of all seleno-DL-methionine supplemented diets tested, and a whole body Se residue of 7.2
mg/kg dw.
The nutritional sufficiency study of Rider et al. (2009) with rainbow trout is unique in that it determined
dietary and whole body selenium requirements for both stressed and unstressed fish. Rider et al. (2009)
observed that rainbow trout stressed by a combination of low water levels in holding tanks and twice
daily handling offish by 30 second aerial exposure in dip nets resulted in a higher nutritional requirement
for selenium than was observed in fish not subjected to the stress routine. They concluded that trout
exposed to physical stressors could benefit from an additional 0.3 - 2.0 mg/kg dw additional selenium
supplementation over and above the Se content of nutritionally Se sufficient diets for fish not undergoing
stress.
The fish with the highest known nutritional requirement for selenium is the non-North American resident
North African catfish (Glorias gariepinus). Abdel-Tawwab et al. (2007) determined in a 12 week study
with fingerlings that Se dietary doses of 1.04 mg/kg dw and 3.67 mg/kg dw were associated with
suboptimal and maximum weight gains of the catfish, respectively. Catfish survival was 100% in both
the Se-deficient and Se-sufficient dietary dose exposures during the 12 week study period. The respective
whole body selenium tissue residues at the end of the 12 week study were 4.72 mg/kg dw in the Se-
deficient fish and 15.43 mg/kg dw in the fish fed the nutritionally sufficient Se diet. North African catfish
(Abdel-Tawwab et al. 2007) is the only known fish species with an identified whole body nutritional
requirement for Se higher than the national aquatic life criterion for whole body Se in fish.
Do not distribute, quote or cite E-59 Draft Document
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Table E-23. Studies with both empirically measured selenium dietary doses and whole body residues associated with nutritional
deficiency and sufficiency in fish.
Species
Malabar grouper
(Epinephelus
malabaricus)
Prussian carp
(Carassius gibelio)
Common carp
(Cyprinus carpio)
Atlantic cod (Gadus
morhua)
Cobia (Rachycentron
canadum)
Coho salmon
(Oncorhynchus kisutch)
Atlantic salmon (Salmo
solar)
North African catfish
(Glorias gariepinus)
Rainbow trout
(Oncorhynchus mykiss)
Lifestage /
Size Wet
wt
Juvenile
12.2 g
Juvenile
2.74 g
Juvenile
26.9 g
Larvae 0.16
g
(estimated
from dry wt
of larvae
Juvenile
6.27 g
Smolt 22.7
g
Fry 0.1 g
Fingerling
68.6 g
Juvenile
0.6 g
Exposure
duration
8 weeks
100 days
120 days
23 days
10 weeks
Hatchery
reared
4 weeks
12 weeks
16 weeks
Ingested
dietary dose
Se mg/kg
dry wt.
0.21
0.47
0.04
1.4
0.21-0.62
0.7-0.9
0.03 -0.04
1.04
0.6-6.6
Se chemical
form
Basal diet
Seleno-
methionine
Basal diet
Basal diet
0.21= Basal
diet, 0.62 =
seleno-DL-
methionine
Not given
Basal diet
Organic Se
Selenite
Na2SeO3-5H2O
Whole
body Se
mg/kg
dry wt
0.64
1.0
1.04
1.1
1.13-
2.11
1.974
2.7
4.72
0.2-1.0
Deficiency
or
Sufficiency
Deficiency
Deficiency
Deficiency
Deficiency
Deficiency
Deficiency
Deficiency
Deficiency
Sufficiency
Deficiency symptoms
Basis for sufficiency
determination
Suboptimal weight gain
and feed efficiency
Suboptimal growth,
feeding rate and feed
conversion rate
Reduced growth and
survival
Larval survival 32%
lower compared to larvae
fed selenium-enriched
diet
Statistically significantly
reduced specific growth
rate and survival
Survival of hatchery
reared smolts 1.5 - 2. Ox
lower than wild smolts
Decreased survival
relative to fry fed diet
supplemented with 0. 1
ug/g Se and 0.5 lU/g
vitamin E
Suboptimal weight gain
and specific growth rate
No deficiency or toxicity
signs on growth
Reference
Lin and Shiau
2005
Han et al.
2011
Gaber2007
Harare et al.
2008
Liu et al.
2010
Felton et al.
1990
Poston et al.
1976
Abdel-
Tawwab et al.
2007
Hilton and
Hodson 1983
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E-60
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Species
Atlantic salmon (Salmo
salar)
Rainbow trout
(Oncorhynchus mykiss)
Malabar grouper
(Epinephelus
malabaricus)
Atlantic salmon (Salmo
salar)
Common carp
(Cyprinus carpio)
Rainbow trout
(Oncorhynchus mykiss)
Prussian carp
(Carassius gibelio)
Hybrid striped bass
(wiper, Morone
chrysops x Morone
saxatilis)
Atlantic salmon (Salmo
salar)
Cobia (Rachycentron
canadum)
Lifestage /
Size Wet
wt
Parr 4.5 g
Juvenile
26.3 g
Juvenile
12.2 g
Parr 4.5 g
Juvenile
26.9 g
Juvenile
26.3 g
Juvenile
2.74 g
Juvenile
2.94 g
Parr 4.5 g
Juvenile
6.27 g
Exposure
duration
8 weeks
1 1 weeks
8 weeks
8 weeks
120 days
1 1 weeks
100 days
12 weeks
8 weeks
10 weeks
Ingested
dietary dose
Se mg/kg
dry wt.
1.2
0.77
0.77
3.4
0.24-0.32
2.3-3.9
1.23-2.77
0.10
3.1
0.85 - 1.36
Se chemical
form
Basal diet
Basal diet
Seleno-
methionine
Selenite
Na2SeO3-5H2O
Selenite
Na2SeO3-5H2O
Selenite
Na2SeO3-5H2O
Seleno-
methionine
Basal diet
Seleno-
methionine
Seleno-DL-
methionine
Whole
body Se
mg/kg
dry wt
0.58-
0.70
0.9
0.92
1.13
1.23 -
1.29
1.6-2.8
1.7-3.4
2.01
2.06
2.58-
2.62
Deficiency
or
Sufficiency
Sufficiency
Sufficiency
Sufficiency
Sufficiency
Sufficiency
Sufficiency
Sufficiency
Sufficiency
Sufficiency
Sufficiency
Deficiency symptoms
Basis for sufficiency
determination
No deficiency signs on
growth, survival or
glutathione peroxidase
activity
Optimal growth, survival
and antioxidant status
Maximal weight gain and
feed efficiency
No deficiency signs on
growth, survival or
glutathione peroxidase
activity
Maximal growth and
survival
Optimal growth, survival
and antioxidant status
Maximal growth, no
effect on survival, no
increase in oxidative
stress
Minimum dietary
requirement for
acceptable survival and
growth
No deficiency signs on
growth, survival or
glutathione peroxidase
activity
Maximal and statistically
identical specific growth
rate and survival
Reference
Lorentzen et
al. 1994
Rider et al.
2009
Lin and Shiau
2005
Lorentzen et
al. 1994
Gaber2007
Rider et al.
2009
Han et al.
2011
Jaramillo
2006
Lorentzen et
al. 1994
Liu et al.
2010
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Species
Rainbow trout
(Oncorhynchus mykiss)
Atlantic cod (Gadus
morhua)
Coho salmon
(Oncorhynchus kisutch)
Hybrid striped bass
(wiper, Morone
chrysops x Morone
saxatilis)
Hybrid striped bass
(wiper, Morone
chrysops x Morone
saxatilis)
North African catfish
(Glorias gariepinus)
Lifestage /
Size Wet
wt
Juvenile
26.3 g
Larvae 0.16
g
(estimated
from dry wt
of larvae
Smolt
14.28 g
Juvenile
2.94 g
Juvenile
2.92 g
Fingerling
68.6 g
Exposure
duration
1 1 weeks
23 days
Wild
smolts
12 weeks
12 weeks
12 weeks
Ingested
dietary dose
Se mg/kg
dry wt.
2.4-4.1
4.8
Se in natural
diet unknown
1.19
0.90
3.67
Se chemical
form
Organic Se -
yeast
Selenite
Na2SeO3-5H2O
Unknown
Selenite
Na2SeO3-5H2O
Seleno-DL-
methionine
Organic Se
Whole
body Se
mg/kg
dry wt
2.8-4.8
3.5
3.63
5.13
7.2
15.43
Deficiency
or
Sufficiency
Sufficiency
Sufficiency
Sufficiency
Sufficiency
Sufficiency
Sufficiency
Deficiency symptoms
Basis for sufficiency
determination
Optimal growth, survival
and antioxidant status
Larval survival increased
32%, growth essentially
unchanged relative to
survival of larvae fed
basal diet
Survival of wild smolts
1.5 - 2. Ox higher than
hatchery reared smolts
Highest weight gain of
any selenite diet test,
significantly higher than
basal diet weight gain
Highest survival and
weight gain of any
seleno-DL-methionine
diet tested
Maximal weight gain,
specific growth rate and
survival
Reference
Rider et al.
2009
Harare et al.
2008
Felton et al.
1990
Jaramillo
2006
Jaramillo
2006
Abdel-
Tawwab et al.
2007
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E-62
Draft Document
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APPENDIX F: TOXICITY OF SELENIUM TO AQUATIC
PLANTS
Do not distribute, quote, or cite F-l Draft Document
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SELENITE
Data are available on the toxicity of selenite to 13 species of freshwater algae and plants (Table F-l).
Results ranged from an LC50 of 70,000 ug/L for the green alga, Chlorella ellipsoidea (Shabana and El-
Attar 1995) to 522 ug/L for incipient inhibition of the green alga, Scenedesmus quadricauda (Bringmann
and Kuhn 1977a, 1978a,b, 1979, 1980b). Foe and Knight (Manuscript) found that 75 ug/L decreased the
dry weight of Selenastrum capricornutum (Table F-l). Wehr and Brown (1985) reported that 320 ug/L
increased the growth of the alga Chrysochromulina breviturrita.
The 96-hr EC50 for the saltwater diatom, Skeletonema costatum, is 7,930 ug/L, based on reduction in
chlorophyll a (Table F-l). Growth of Chlorella sp., Platymonas sub cordifor mis, and Fucus spiralis
increased at selenite concentrations from 2.6 to 10,000 ug/L (Table F-l). Other marine algae exposed to
selenite from 14 to 60 days had no observed effect concentrations (NOAEC) that ranged from 1,076 to
107,606 ug/L. These data suggest that saltwater plants will not be adversely affected by concentrations of
selenite that do not affect saltwater animals.
SELENATE
Growth of several species of green algae was affected by concentrations ranging from 100 to 40,000 ug/L
(Table F-l). Blue-green algae appear to be more tolerant to selenate with 1,866 ug/L being the lowest
concentration reported to affect growth (Kiffney and Knight 1990). Kumar (1964) found that a blue-green
alga developed and lost resistance to selenate. The difference in the sensitivities of green and blue-green
algae to selenate might be of ecological significance, particularly in bodies of water susceptible to
nuisance algal blooms. For example, Patrick et al. (1975) reported that a concentration of 1,000 ug/L
caused a natural assemblage of algae to shift to a community dominated by blue-green algae.
The saltwater coccolithophore, Cricosphaera elongata, had reduced growth when exposed to 41,800 ug/L
selenate for 14 days (Boisson et al. 1995). Seven other saltwater algal species investigated by Wong and
Oliveira (1991a) exhibited NOEC growth values that ranged from 1,043 to 104,328 ug/L. At 10,000 ug/L,
selenate is lethal to four species of saltwater phytoplankton and lower concentrations increase or decrease
growth (Table F-l). Wheeler et al. (1982) reported that concentrations as low as 10 ug/L reduced growth
of Porphyridium cruentum (Table F-l).
Although selenite appears to be more acutely toxic than selenate to most aquatic animals, this does not
seem to be true for aquatic plants. Selenite and selenate are about equally toxic to the freshwater algae
Do not distribute, quote, or cite F-2 Draft Document
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Anabaena cylindrica, Anabaena flos-aquae, Anabaena variabilis, Anacystis nidulans, and Scenedesmus
dimorphus (Kiffhey and Knight 1990; Kumar and Prakash 1971; Moede et al. 1980) and the saltwater
algae Agemenellum quadroplicatum, Chaetoceros vixvisibilis and Amphidinium carterae (Wong and
Oliveira 1991a). The two oxidation states equally stimulated growth of Chrysochromulina breviturrita
(Wehr and Brown 1985). On the other hand, selenate is more toxic than selenite to the freshwater
Selenastrum capricornutum (Richter 1982; Ibrahim and Spacie 1990) and the saltwater Chorella sp.,
Platymonas subcordiformis and Nannochloropsis oculata (Wheeler et al. 1982; Wong and Oliveira
1991a). In addition, Fries (1982) found that growth of thalli of the brown macroalga, Fucus spiralis, was
stimulated more by exposure to selenite at 2.605 ug/L than to the same concentration of selenate.
A Final Plant Value, as defined in the Guidelines, cannot be obtained because no test in which the
concentrations of selenite or selenate were measured and the endpoint was biologically relevant has been
conducted with an important aquatic plant species.
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Table F-l. Toxicity of Selenium to Aquatic Plants
Species
Green alga,
Chlorella vulgaris
Green alga,
Chlorella
ellipsoidea
Green alga,
Scenedesmus
dimorphus
Green alga,
Scenedesmus
quadricauda
Green alga,
Scenedesmus
quadricauda
Green alga,
Selenastrum
capricornutum
Green alga,
Selenastrum
capricornutum
Blue-green alga,
Anabaena
constricta
Blue-green alga,
Anabaena
cylindrica
Blue-green alga,
Anabaenaflos-
aquae
Blue-green alga,
Anabaena
variabilis
Blue-green alga,
Anacystis nidulans
Blue-green alga,
Microcystis
aeruginisa
Alga,
Euglena gracilis
Chemical
Sodium
selenite
Sodium
selenite
Sodium
selenite
Sodium
selenite
Sodium
selenite
Sodium
selenite
Sodium
selenite
Sodium
selenite
Sodium
selenite
Sodium
selenite
Sodium
selenite
Sodium
selenite
Sodium
selenite
_
Hardness
(mg/L as Duration
CaCO^ (days) Effect
FRESHWATER SPECIES
Selenium (IV)
90-120 Reduced
growth
7 EC50
14 Reduced
growth
8 Incipient
inhibition
8 Incipient
inhibition
4 EC50
6 EC50
7 EC50
14 Reduced
growth
10 Reduced
chlorophyll a
6-18 LC50
10-18 LC50
8 Incipient
inhibition
1 5 Reduced
growth
Concentration
rug/L)a
5,480
70,000
24,000
522
2,500
2,900
65,000
67,000
24,000
1,866
15,000b
30,000b
9,400
(9,300)
5,920
Reference
De Jong 1965
Shabana and El-
Attar 1995
Moede et al.
1980
Bringmann and
Kuhn 1977a;
1978a,b; 1979;
1980b
Bringmann and
Kuhn 1959a
Richterl982
Ibrahim and
Spacie 1990
Shabana and El-
Attar 1995
Moede et al.
1980
Kiffney and
Knight 1990
Kumar and
Prakash 1971
Kumar and
Prakash 1971
Bringmann and
Kuhn 1976;
1978a,b
Bariaud and
Mestre 1984
Do not distribute, quote, or cite
F-4
Draft Document
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Species
Duckweed,
Lemna minor
Duckweed,
Lemna minor
Duckweed,
Lemna minor
Green alga,
Ankistrodesmus
falcatus
Green alga,
Scenedesmus
dimorphus
Green alga,
Scenedesmus
obliquus
Green alga,
Selenastrum
capricornutum
Green alga,
Selenastrum
capricornutum
Green alga,
Selenastrum
capricornutum
Blue-green alga,
Anabaena
cylindrica
Blue-green alga,
Anabaenaflos-
aquae
Blue-green alga,
Anacystis nidulans
Blue-green alga,
Anabaena
viriabilis
Blue-green alga,
Microcoleus
vaginatus
Duckweed,
Lemna minor
Chemical
_
Sodium
selenite
Sodium
selenite
Sodium
selenate
Sodium
selenate
Sodium
selenate
Sodium
selenate
Sodium
selenate
Sodium
selenate
Sodium
selenate
Sodium
selenate
Sodium
selenate
Sodium
selenate
Sodium
selenate
Sodium
selenate
Hardness
(mg/L as Duration
_CaCO3} (days) Effect
4 EC50
14 EC50
(mult, rate)
14 NOEC
(mult, rate)
Selenium (VI)
14 Did not
reduce
growth
14 Reduced
growth
14 Reduced
growth
14 Reduced
growth
4 EC50
6 EC50
14 Reduced
growth
10 Reduced
chlorophyll a
6-18 EC50
10-18 EC50
14 Reduced
growth
14 EC50
(mult, rate)
Concentration
rug/L)a
2,400
3,500
800
10
22,100
100
300
199
<40,000
22,100
1,866
39,000b
17,000b
10,000
11,500
Reference
Wang 1986
Jenner and
Janssen-
Mommen 1 993
Jenner and
Janssen-
Mommen 1 993
Vocke et al.
1980
Moede et al.
1980
Vocke et al.
1980
Vocke et al.
1980
Richter 1982
Ibrahim and
Spacie 1990
Moede et al.
1980
Kiffney and
Knight 1990
Kumar and
Prakash 1971
Kumar and
Prakash 1971
Vocke et al.
1980
Jenner and
Janssen-
Mommen 1 993
Do not distribute, quote, or cite
F-5
Draft Document
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Hardness
Species
Duckweed,
Lemna minor
Species
Green alga,
Dunaliella
tertiolecta
Cyanophyceae alga,
Agemenellum
quadruplicatum
Diatom,
Chaetoceros
vixvisibilis
Diatom,
Skeletonema
costatum
Coccolithophore,
Cricosphaera
elongata
Dinoflagellate,
Amphidinium
carterae
Dinoflagellate,
Peridinopsis borgei
Eustigmatophyceae
alga,
Nannochloropsis
oculata
Pyrmnesiophyceae
alga,
Isochrysis galbana
Pyrmnesiophyceae
alga,
Pavlova lutheri
Green alga,
Dwwa/ze//a
ferfzo/ecfa
(mg/L as Duration
Chemical CaCO,) (days) Effect
Sodium
selenate
Chemical
Sodium
selenite
Sodium
selenite
Sodium
selenite
Selenious
acid0
Sodium
selenite
Sodium
selenite
Selenium
oxide
Sodium
selenite
Sodium
selenite
Sodiun selenite
Sodium
selenate
14 NOEC
(mult. Rate)
Salinity Duratio
(2/ks) n Effect
(days)
SALTWATER SPECIES
Selenium (IV)
60 NOEC growth
60 NOEC growth
60 NOEC growth
4 EC50 (reduction
in chlorophyll a)
14 Reduced growth
60 NOEC growth
70-75 Maximum
growth
60 NOEC growth
60 NOEC growth
60 NOEC growth
Selenium (VI)
60 NOEC growth
Concentration
rug/L)a
>2,400
Concentration
rug/L)a
1,076
10,761
1,076
7,930
4,570
10,761
0.01-0.05
107,606
1,076
1,076
104,328
Reference
Jenner and
Janssen-
Mommen 1 993
Reference
Wong and
Oliveiral991a
Wong and
Oliveiral991a
Wong and
Oliveiral991a
U.S. EPA 1978
Boisson et al.
1995
Wong and
Oliveiral991a
Lindstrom 1985
Wong and
Oliveiral991a
Wong and
Oliveiral991a
Wong and
Oliveiral991a
Wong and
Oliveiral991a
Do not distribute, quote, or cite
F-6
Draft Document
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Species
Cyanophyceae alga,
Agemenellum
quadruplicatum
Diatom,
Chaetoceros
vixvisibilis
Coccolithophore,
Cricosphaera
elongata
Dinoflagellate,
Amphidinium
carterae
Eustigmatophyceae
alga,
Nannochloropsis
oculata
Pyrmnesiophyceae
alga,
Isochrysis galbana
Pyrmnesiophyceae
alga,
Pavlova lutheri
Chemical
Sodium
selenate
Sodium
selenate
Sodium
selenate
Sodium
selenate
Sodium
selenate
Sodium
selenate
Sodium
selenate
Salinity Duratio Concentration
(2/ks) n Effect (U2/L)a Reference
(days)
60 NOEC growth
60 NOEC growth
14 Reduced growth
60 NOEC growth
60 NOEC growth
60 NOEC growth
60 NOEC growth
10,433 Wong and
Oliveiral991a
1,043 Wong and
Oliveiral991a
41,800 Boissonetal.
1995
10,433 Wong and
Oliveiral991a
10,433 Wong and
Oliveiral991a
10,433 Wong and
Oliveiral991a
104,328 Wong and
Oliveiral991a
' Concentration of selenium, not the chemical.
b Estimated from published graph.
0 Reported by Barrows et al. (1980) in work performed under the same contract.
Do not distribute, quote, or cite
F-7
Draft Document
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APPENDIX G: UNUSED DATA
Do not distribute, quote, or cite G-l Draft Document
-------
Based on the requirements set forth in the guidelines (Stephen et al. 1985) the following studies are not
acceptable for the following reasons and are classified as unused data. Note the acceptance of chronic
toxicity data included diet and field exposures where selenium was the dominant toxicant.
Studies Were Conducted with Species That Are Not Resident in North America
Ahsanullah and Brand (1985) Gotsis (1982) Ringdal and Julshamn (1985)
Ahsanullah and Palmer Hiraika et al. (1985) Rouleau et al. (1992)
(1980) Juhnke and Ludemann (1978) Sastry and Shukla (1994)
Baker and Davies (1997) Kitamura(1990) Savant and Nilkanth (1991)
Barghigiani et al. (1993) Manoharan and Prabakaran Shultz and Ito (1979)
Chidambaram and Sastry (1994) Srivastava and Tyagi (1985)
(1991a,b) Minganti et al. (1994, 1995) Takayanagi (2001)
Congiuetal. (1989) Niimi and LaHam (1975, Tomasik et al. (1995b)
Cuvin and Furness (1988) 1976) Tian and Liu (1993)
Fowler and Benayoun Regoli (1998) Wrench (1978)
(1976a,b) Regoli and Principato (1995)
Gaikwad (1989) Rhodes et al. (1994)
Deelstra et al. (1989), Forsythe and Klaine (1994), Okasako and Siegel (1980) and Petrucci et al. (1995)
conducted tests with brine shrimp species that are too atypical to be used in derving national criteria.
These Studies or Reviews Contain Relevant Data That Have Been Published Elsewhere
Adams and Johnson (1981) Eisler (1985) McKee and Wolf (1963)
Biddinger and Gloss (1984) Hall and Burton (1982) National Research Council
Bowie et al. (1996) Hodson and Hilton (1983) (1976) Neuhold (1987)
Brandao et al. (1992) Hodson et al. (1984) NCDNR&CD (1986)
Brooks (1984) Jenkins (1980) Peterson and Nebeker (1992)
Burton and Stemmer (1988) Kaiser et al. (1997) Phillips and Russo (1978)
Chapman et al. (1986) Kay (1984) Presser (1994)
Davies (1978) LeBlanc (1984) Roux et al. (1996)
Debruyn and Chapman Lemly (1993c, 1996ab, Swift (2002)
(2007) 1997d) Thompson et al. (1972)
Devillers et al. (1988) Lemly and Smith (1987) Versar (1975)
Do not distribute, quote, or cite G-2 Draft Document
-------
Authors Did Not Specify the Oxidation State of Selenium Used in Study
Greenberg and Kopec (1986) Kapu and Schaeffer (1991)
Hutchinson and Stokes Kramer et al. (1989) Rauscher (1988)
(1975) Mahan et al. (1989) Snell et al. (1991b)
Not Useful Because of No Effects Observed at Exposure Concentrations or Insufficient Number of
Treatments
Muscatello and Janz (2009)
Pyle et al. (2005)
Schlenk et al (2003)
Chronic Study with no Dietary Exposure
Hopkins et al. (2002)
Oti (2005)
Rowe (2003)
Teh et al. (2002)
Selenium Was a Component
Apteetal. (1987)
Baeretal. (1995)
Baker etal. (1991)
Berg etal. (1995)
Besseretal. (1989)
Biedlingmaier and Schmidt
(1989)
Bjoernberg(1989)
Bjoernberg etal. (1988)
Bleckmann etal. (1995)
Boisson etal. (1989)
Bondavalli etal. (1996)
Bowmeretal. (1994)
Briegeretal. (1992)
Burton and Pinkney (1984)
Burton etal. (1983, 1987a)
of an Effluent, Fly Ash, Formulation,
Cherry etal. (1987)
Cieminski and Flake (1995)
Clark etal. (1989)
Cooke and Lee (1993)
Cossu etal. (1997)
Coyle etal. (1993)
Crane etal. (1992)
Crock etal. (1992)
Cushman etal. (1977)
Davies and Russell (1988)
dePeysteretal. (1993)
Dickman and Rygiel (1996)
Dierenfeldetal. (1993)
Doebel et al. (2004)
Drndarski etal. (1990)
Mixture, Sediment or Sludge
Eriksson and Forsberg (1992)
Eriksson and Pedros-Alio
(1990)
Fairbrotheretal. (1994)
Favaetal. (1985a,b)
Feroci etal. (1997)
Finger and Bulak (1988)
Finley(1985)
Fisher and Wente (1993)
Fjeld and Rognerud (1993)
Fletcher etal. (1994)
Follett(1991)
Gerhardt(1990)
Gerhardtetal. (1991)
Gibbs and Miskiewicz (1995)
Graham etal. (1992)
Do not distribute, quote, or cite
G-3
Draft Document
-------
Gunderson et al. (1997)
Hall (1988)
Hall etal. (1984, 1987,
1988,1992)
Hamilton et al. (1986, 2000)
Harrison etal. (1990)
Hartwell et al. (1987ab, 1988,
1997)
Hatcher etal. (1992)
Haynes etal. (1997)
Haywardetal. (1996)
Hellouetal. (1996b)
Henebry and Ross (1989)
Henny etal. (1989, 1990,
1995)
Hildebrandetal. (1976)
Hjeltnes and Julshman (1992)
Hockett and Mount (1996)
Hodson(1990)
Hoffman etal. (1988, 1991)
Homziak etal. (1993)
Hopkins et al. (2000)
Hopkins et al. (2004)
Hothem and Welsh (1994a)
Jackson (1988)
Jackson etal. (1990)
Jacquezetal. (1987)
Jay and Muncy (1979)
Jayasekera (1994)
Jayasekera and Rossbach
(1996)
Jenner and Bowmer (1990)
(1992)
Jin etal. (1997)
Jorgensen and Heisinger
(1987)
Karlson and Frankenberger
(1990)
Kemble etal. (1994)
Kennedy (1986)
Kerstenetal. (1991)
King and Cromartie (1986)
King etal. (1991, 1994)
Kluseketal. (1993)
Koh and Harper (1988)
Koike etal. (1993)
Krishnajaetal. (1987)
Kruuk and Conroy (1991)
Kuehl and Haebler( 1995)
Kuehl etal. (1994)
Kuss etal. (1995)
Landau etal. (1985)
Livingston et al. (1991)
Lobel etal. (1990)
Luoma and Phillips (1988)
Lundquist etal. (1994)
Lyle (1986)
MacFarlane etal. (1986)
Mann and Fyfe (1988)
Marcogliese et al. (1987)
Marvin and Howell.
(1997)Maureretal (1999)
McCloskey and Newman
(1995)
McCloskey etal. (1995)
McCrea and Fischer (1986)
McLean etal. (1991)
Mehrle etal. (1987)
Metcalf-Smith (1994)
Micallef and Tyler (1989)
Mikac etal. (1985)
Miles and Tome (1997)
Miller etal. (1996)
Misitano and Schiewe (1990)
Moore (1988)
Munawar and Legner (1993)
Muskettetal. (1985)
Naddy etal. (1995)
Nielsen and Bjerregaard
(1991)
Norman etal. (1992)
Nuutinen & Kukkonen
(1998)
Oberbach and Hartfiel (1987,
1988)
Oberbach etal. (1989)
Ohlendorf etal. (1989, 1990,
1991)
Olson and Welsh (1993)
Peters etal.(1999)
Phillips and Gregory (1980)
Pratt and Bowers (1990)
Presserand Ohlendorf (1987)
Prevot and Sayer-Gobillard
(1986)
Pritchard(1997)
Pyle etal. (2001)
Reash et al. (1988, in press)
Rhodes and Burke (1996)
Do not distribute, quote, or cite
G-4
Draft Document
-------
Ribeyre et al. (1995) Sorenson and Bauer (1983) Weres et al. (1990)
Rice et al. (1995) Specht et al. (1984) White and Geitner (1996)
Riggs and Esch (1987) Steele et al. (1992) Wiemeyer et al. (1986)
Riggs et al. (1987) Stemmer et al. (1990) Wildhaber and Schmitt
Robertson et al. (1991) Summers et al. (1995) (1996)
Roper et al. (1997) Thomas et al. (1980b) Williams et al. (1989)
Rowe et al. (1996) Timothy et al. (2001) Wolfe et al. (1996)
Russell et al. (1994) Trieff et al. (1995) Wolfenberger (1987)
Ryther et al. (1979) Turgeon and O=Conner Wong and Chau (1988)
Saiki and Jenings (1992) (1991) Wong et al. (1982)
Saiki and Ogle (1995) Twerdok et al. (1997) Wu et al. (1997)
Saleh et al. (1988) Unsal (1987) Yamaoka et al. (1994)
Seelye et al. (1982) Van Metre and Gray (1992) Zagatto et al. (1987)
Sevareid and Ichikawa Wahl et al. (1994) Zaidi et al. (1995)
(1983) Wandan and Zabik (1996) Zhang et al. (1996)
Skinner (1985) Wang et al. (1992, 1995)
Somerville et al. (1987) Welsh (1992)
Exposed enzymes, excised tissue or tissue extractor
Tripathi and Pandey (1985) and Heinz (1993b) used test organisms that had been previously exposed to
pollutants in food or water.
Albers et al. (1996) Bell et al. (1984, 1985, Byl et al. (1994)
Al-Sabti (1994, 1995) 1986a,b, 1987ab) Chandy and Patel (1985)
Arvy et al. (1995) Berges and Harrison (1995) Chen et al. (1997)
Augier et al. (1993a, b) Blondin et al. (1988) Cheng et al. (1993)
Avery et al. (1996) Boisson et al. (1996) Christensen and Tucker
Baatrup (1989) Bottino et al. (1984) (1976)
Baatrup and Dansher (1987) Braddon (1982) Dabbert and Powell (1993)
Baatrup et al. (1986) Braddon-Galloway and DeQuiroga et al. (1989)
Babichetal. (1986, 1989) Balthrop (1985) Dierickx (1993)
Barrington et al. (1997) Bradford et al. (1994a,b) Dietrich et al. (1987)
Becker et al. (1995a,b) Brandt et al. (1990) Dillio et al. (1986)
Do not distribute, quote, or cite G-5 Draft Document
-------
Doyotteetal. (1997)
Drotaretal. (1987)
Dubois and Callard (1993)
Ebringeretal. (1996)
Engberg and Borsting (1994)
Engbergetal. (1993)
Eunetal. (1993)
Foltinova and Gajdosova
(1993)
Foltinova etal. (1994)
Freeman and Sanglang
(1977)
Grubor-Lajsic et al. (1995)
Hait and Sinha( 1987)
Hanson (1997)
Heisinger and Scott (1985)
Heisinger and Wail (1989)
Henderson etal. (1987)
Henny and Bennett (1990)
Hoffman and Heinz (1988,
1998)
Hoffman etal. (1989, 1998)
Hoglund(1991)
Hontelaetal. (1995)
Hsu etal. (1995)
Hsu and Goetz (1992)
Ishikawaetal. (1987)
James etal. (1993)
Jovanovic et al. (1995, 1997)
Kai etal. (1995)
Kedziroski etal. (1996)
Kelly etal. (1987)
Kralj and Stunja( 1994)
Lalitha and Rani (1995)
Lanetal. (1995)
Lemaire etal. (1993)
Livingstone et al. (1992)
Low and Sin (1995, 1996)
Micallef and Tyler (1990)
Montagnese et al. (1993)
Murataetal. (1996)
Nakonieczny (1993)
Neuhierl and Boeck (1996)
Nigro(1994)
Nigro etal (1992)
Norheim and Borch-Iohnsen
(1990)
Norheim etal. (1991)
O=Brienetal. (1995)
Olson and Christensen (1980)
Overbaugh and Fall (1985)
Palmisano et al. (1995)
Patel etal. (1990)
Patel and Chandy (1987)
Perez Campo etal. (1990)
Perez-Trigo etal. (1995)
Phadnis etal. (1988)
Price and Harrison (1988)
Rady etal. (1992)
Rani and Lalitha (1996)
Regoli etal. (1997)
Schmidt etal. (1985)
Schmitt etal. (1993)
Segneretal. (1994)
Sen etal. (1995)
Shigeokaetal. (1990, 1991)
Siwicki etal. (1994)
Srivastava and Srivastava
(1995)
Sun etal. (1995)
Takedaetal. (1992a,b,(1993,
1997)
Treuthardt(1992)
Vazquez etal. (1994)
Veenaetal. (1997)
Wise etal. (1993a,b)
Wong and Oliveira (1991b)
Yokotaetal. (1988)
Test procedures test material or results were not adequately described by Botsford (1997), Botsford et al.
(1997, 1998), Bovee (1978), Gissel-Nielsen and Gissel-Nielsen (1973, 1978), Greenberg and Kopec
(1986), Mauk (2001), and Nassos et al. (1980) or when the test media contained an excessive amount
(>200 ug/L) of EDTA (Riedel and Sanders (1996).
Do not distribute, quote, or cite
G-6
Draft Document
-------
Some data obtained from tests conducted with just one exposure concentration to evaluate acute or
chronic toxicity were not used (e.g., Bennett 1988; Heinz and Hoffman 1998; Munawar et al. 1987;
Pagano et al. 1986; Wolfenberger 1986).
Kaiser (1980) calculated the toxicities of selenium(IV) and selenium(VI) to Daphnia magna based on
physiochemical parameters. Kumar (1964) did not include a control treatment in the toxicity tests. The
daphnids were probably stressed by crowding in the tests reported be Schultz et al. (1980). Siebers and
Ehlers (1979) exposed too few test organisms as did Owsley (1984) in some tests.
Selenium Concentrations Reported in Wild Aquatic Organisms Were Insufficient to Calculate BAF
Abdel-Moati and Atta (1991)
Adeloju and Young (1994)
Aguirreetal. (1994)
Akesson and Srikumar
(1994)
Aksnesetal. (1983)
Allen and Wilson (1990)
Ambulkar et al. (1995)
Amiardetal. (1991, 1993)
Andersen and Depledge
(1997)
Andreev and Simeonov
(1992)
Angulo (1996)
Arrulaetal. (1996)
Arway (1988)
Ashton(1991)
Augieretal. (1991, 1993,
1995a,b)
Augspurger et al. (1998)
Averyetal. (1996)
Badsha and Goldspink (1988)
Baines and Fisher (2001)
Baldwin and Maher (1997)
Baldwin etal. (1996)
Barghigiani(1993)
Barghigiani et al. (1991)
Baron etal. (1997)
Batley(1987)
Baumann and Gillespie
(1986)
Baumann and May (1984)
Beal (1974)
Beck etal. (1997)
Belandetal. (1993)
Beliaeffetal. (1997)
Bell and Cowey (1989)
Benemariya et al. (1991)
Berry etal. (1997)
Bertram etal. (1986)
Besseretal. (1994, 1993)
Birkner(1978)
Boisson and Romeo (1996)
Bowerman et al. (1994)
Braune eta. (1991)
Brezina and Arnold (1977)
Brugmann and Hennings
(1994)
Brugmann and Lange (1988)
Brumbaugh and Walther
(1991)
Burger (1992, 1994, 1995,
1996, 1997a,b)
Burger and Gochfeld
(1992a,b, 1993, 1995 ab,
1996, 1997)
Burger etal. (1992a,b,c,1993,
1994a,b)
Byrne and DeLeon (1986)
Byrne etal. (1985)
Cantillo etal. (1997)
Capar and Yess (1996)
Capelli etal. (1987, 1991)
Cappon (1984)
Cappon and Smith (1981)
(1982a,b)
Cardellicchio (1995)
Carell etal. (1987)
Carter and Porter (1997)
Do not distribute, quote, or cite
G-7
Draft Document
-------
Caurantetal. (1994, 1996)
Chau and Riley (1965)
Chiang etal. (1994)
Chou and Uthe (1991)
Chvojka(1988)
Chvojkaetal. (1990)
Clifford and Harrison (1988)
Collins (1992)
Combs etal. (1996)
Cosson etal. (1988)
Courtney etal. (1994)
Cruwys etal. (1994)
Crutchfield (2000)
Cumbie and Van Horn (1978)
Currey etal. (1992)
Custer and Hohman (1994)
Custer and Mitchell (1991,
1993)
Custer etal. (1997)
Dabeka and McKenzie
(1991)
Davoren(1986)
Deaker and Maher (1997)
Demon etal. (1988)
Dietz etal. (1995, 1996)
Doherty etal. (1993)
Elliott and Scheuhammer
(1997)
Eriksson etal. (1989)
Evans etal. (1993)
Feltonrtal. (1990)
Felton etal. (1994)
Fitzsimons et al. (1995)
Focardi etal. (1985, 1988)
Fowler (1986)
Fowler etal. (1975, 1985)
France(1987)
Friberg(1988)
Froslie etal. (1985, 1987)
Gabrashanske and Daskalova
(1985)
Gabrashanska and Nedeva
(1994)
Galgan and Frank (1995)
Garcia - Hermandez et al.
(2000)
Giardinaetal. (1997)
Gillespie and Baumann
(1986)
Gochfeld (1997)
Goede (1985, 1991, 1993a,b)
Goede etal. (1989, 1993)
Goede and DeBruin (1984,
1985)
Goede and Wolterbeek
(1993, 1994a,b)
Gras etal. (1992)
Greig and Jones (1976)
Gutenmann et al. (1988)
Gutierrez-Galindo et al.
(1994)
Guvenetal. (1992)
Halbrooketal. (1996)
Hall and Fisher (1985)
Hamilton and Waddell
(1994)
Hamilton and Wiedmeyer
(1990)
Hansenetal. (1990)
Hardiman and Pearson
(1995)
Hargrave etal. (1992)
Harrison and Klaverkamp
(1990)
Hasunuma et al. (1993)
Haynes etal. (1995)
Hein etal. (1994)
Heiny and Tate (1997)
Heinz (1993a)
Heinz and Fitzgerald
(1993a,b)
Heit(1985)
Heit and Klusek (1985)
Heitetal. (1980, 1989)
Hellouetal. (1992a,b)
(1996a,b)
Henny and Herron (1989)
Hodge etal. (1996)
Hilton etal. (1982)
Honda etal. (1986)
Hothem and Ohlendorf
(1989)
Hothem and Welsh (1994b)
Hothem and Zador( 1995)
Hothem etal. (1995)
Houptetal. (1988)
Hunter etal. (1995, 1997)
Ibrahim and Farrag (1992)
Ibrahim and Mat (1995)
Ishikawaetal. (1993)
Itano etal. (1984, 1985a,b)
Jarman etal. (1996)
Do not distribute, quote, or cite
G-8
Draft Document
-------
Johns etal. (1988)
Johnson (1987)
Jop etal. (1997)
Jorhem etal. (1994)
Julshamn etal. (1987)
Kaietal. (1986a,b, 1988,
1992a,b, 1996)
Kaiser etal. (1979)
Kalas etal. (1995)
Kidwell etal. (1995)
Koemanetal. (1973)
Kovacs etal. (1984)
Krogh and Scanes(1997)
Krushevskaetal. (1996)
Lakshmanan and Stephen
(1994)
Lalithaetal. (1994)
LamLeung etal. (1991)
Lanetal. (1994a,b)
Langlois and Langis (1995)
Larsen and Stuerup (1994)
Larsen etal. (1997)
Lauchli (1993)
Law etal. (1996)
Lee and Fisher (1992a,b,
1993)
Leighton and Wobeser
(1994)
Leland and Scudder (1990)
Lemly (1985a, 1994)
Leonzio etal. (1986, 1989,
1992)
Leskinen etal. (1986)
Li etal. (1996)
Lie etal. (1994)
Liu etal. (1987)
Lizamaetal. (1989)
Lobeletal. (1989, 1991,
1992a,b)
Lonzarich etal. (1992)
Lourdes etal. (1990)
Lowe etal. (1985)
Lucas etal. (1970)
Lytle and Lytle (1982)
Mackey etal. (1996)
Maher(1987)
Maheretal. (1992, 1997)
Mann etal. (1988)
Mason et al. (2000)
Masuzawaetal. (1988)
Matsumoto (1991)
Mavenetal. (1995)
May and McKinney (1981)
Mcdowell etal. (1995)
McKenzie-Parnell et al.
(1988)
Meadoretal. (1993)
Mehrle etal. (1982)
Meltzeretal. (1993)
Metcalfe-Smith et al. (1992,
1996)
Michotetal. (1994)
Mills etal. (1993)
Moharram etal. (1987)
Moller(1996)
Mora and Anderson (1995)
Moreraetal. (1997)
Muiretal. (1988)
Mutanenetal. (1986)
Nadkarni and Primack (1993)
Nakamoto and Hassler
(1992)
Narasaki and Cao (1996)
Navarrete et al. (1990)
Nettletonetal. (1990)
Nicola etal. (1987)
Nielsen and Dietz (1990)
Norheim(1987)
Norheim etal. (1992)
Norrgren et al. (1993)
Norstrom etal. (1986)
O=Conner(1996)
O=Sheaetal. (1984)
Oberetal. (1987)
Oehlenschlager(1997)
Ohlendorf(1986)
Ohlendorf and Harrison
(1986)
Ohlendorf and Marois (1990)
Ohlendorf et al. (1986a,b,
1987, 1988a,b)
Okazaki and Panietz (1981)
Ostapczuketal. (1997)
Pakkalaetal. (1972)
Pal et al. (1997) Palawski et
al. (1991)
Palmer-Locarnini and Presley
(1995)
Paludan-Miller et al. (1993)
Papadopoulou and Andreotis
(1985)
Park and Presley (1997)
Do not distribute, quote, or cite
G-9
Draft Document
-------
Park etal. (1994)
Paveglioetal. (1994)
Payer and Runkel (1978)
Payer etal. (1976)
Pennington etal. (1982)
Presley etal. (1990)
Quevauvilleretal. (1993a,b)
Ramos etal. (1992)
Rao etal. (1996)
Reinfelder and Fisher (1991)
Reinfelderetal. (1993, 1998)
Renzoni etal. (1986)
Rigetetal. (1996)
Risenhoover(1989)
Roditi (2000)
Rouxetal. (1994)
Ruelle and Keenlyne (1993)
Sager and Cofield (1984)
Saiki(1986ab, 1987, 1990)
Saiki and Lowe (1987)
Saiki and May (1988)
Saiki and Palawski (1990)
Saiki etal. (1992, 1993)
Sanders and Gilmour (1994)
Scanes(1997)
Scheuhammer et al. (1988)
Schantzetal. (1997)
Schmitt and Brumbaugh
(1990)
Schramel and Xu( 1991)
Schuleretal. (1990)
Scott and Latshaw (1993)
Secoretal. (1993)
Seelye etal. (1982)
Sharif etal. (1993)
Shenetal. (1997)
Shirasaki etal. (1996)
Shultz and Ito (1979)
Simopoulos (1997)
Skaare etal. (1990, 1994)
Smith and Flegal (1989)
Smith etal. (1992)
Sorensen (1988)
Sorensen and Bauer
(1984a,b) Sorensen and
Bjerregaard(1991)
Sorensen etal. (1982, 1983,
1984)
Southworth et al. (2000)
Sparling and Lowe (1996)
Speyer(1980)
Steimle etal. (1994)
Stoeppleretal. (1988)
Stone etal. (1988)
Stripp etal. (1990)
Sundarrao etal. (1991)
(1992)
Svensson et al. (1992)
Tabakaetal. (1996)
Talbot and Chang (1987)
Tallandini etal. (1996)
Tan and Marshall (1997)
Tang etal. (1997)
Tao etal. (1993)
Teherani (1987)
Teigenetal. (1993)
Thomas etal. (1999)
Tilbury etal. (1997)
Topcuoglu etal. (1990)
TranVan and Teherani (1988)
Trocine and Trefry (1996)
Uthe and Bigh (1971)
Vanderstoep et al. (1990)
Varanasi etal. (1993, 1994)
Vitaliano and Zdanowicz
(1992)
Vlieg (1990)
Vlieg etal. (1993)
Vosetal. (1986)
Waddell and May (1995)
Wagemann(1988)
Wagemann and Stewart
(1994)
Wagemann etal. (1988)
(1996) Walsh etal. (1977)
Wang (1996)
Ward and Flick (1990)
Warren etal. (1990)
Weber (1985)
Welsh and Maughan (1994)
Wen etal. (1997)
Wenzel and Gabrielsen
(1995)
Whyte and Boutillier (1991)
Williams etal. (1994)
Wilson etal. (1992, 1997)
Winger and Andreasen
(1985)
Winger etal. (1984, 1990)
Woock and Summers (1984)
Wren etal. (1987)
Wu and Huang (1991)
Do not distribute, quote, or cite
G-10
Draft Document
-------
Yamaoka et al. (1996) Yoshida and Yasumoto Zeisler et al. (1988, 1993)
Yamazaki et al. (1996) (1987) Zhou and Liu (1997)
Zattaetal. (1985)
Do not distribute, quote, or cite G-ll Draft Document
-------
APPENDIX H: CALCULATION OF EF VALUES
Do not distribute, quote, or cite H-l Draft Document
-------
The EPA calcuated EF values by searching its database of selenium measurements and identifying all the selenium measurements from algae,
detritus, or sediment. The EPA then searched for corresponding water column measurements from samples collected at the same aquatic site
within one year of the particulate sample. If more than one water measurement was available for any given particulate measurement, the median
was used. For each of these matched pairs of particulate and water measurements, the EPA calculated the ratio of particulate concentration to
water concentration. If more than one ratio for any given category of particulate material (algae, detritus, or sediment) was calculated at an aquatic
site, the EPA used the median ratio. The geometric mean of the algae, detritus, and sediment ratios was used as the site EF. Because there were at
most only 3 possible values (one for algae, one for detritus, and one for sediment), the EPA used the geometric mean in order to reduce the
potential for one of the values to have excessive influence on the final site EF value. Sites with insufficient data to fulfill these criteria are left
blank.
The EPA evaluated differences in bioaccumulation between different categories of aquatic systems by analyzing EF values for different categories.
The EPA sequentially consolidated categories and examined differences in the distribution of EF values between categories. See text for a
complete description of this analysis.
Reference
Birkner 1978
Birkner 1978
Birkner 1978
Birkner 1978
Birkner 1978
Birkner 1978
Birkner 1978
Butler et al.
1991
Butler et al.
1993
Butler et al.
1993
Do not distribute,
Site description
Meeboer Lake, Laramie
WY
Sweltzer Lake, Delta CO
Galett Lake, Laramie WY
Twin Butter Reservoir,
Laramie WY
Larimer Highway 9 Pond,
Fort Collins CO
East Allen Reservoir,
Medicine Bow WY
Miller's Lake, Wellington
CO
Uncompahgre River at
Colona
Navajo Reservoir, Piedra
River Arm, near La Boca
Spring Cr. at La Boca
quote, or cite
Site ID
3
27
7
23
30
20
22
4
N2
SP2
Specific
waterbody
type-
original
Lake
Lake
Lake
Reservoir
Pond
Reservoir
Lake
River
Reservoir
Creek
Specific
waterbody
type-
Lentic or
Lotic
Lentic
Lentic
Lentic
Lentic
Lentic
Lentic
Lentic
Lotic
Lentic
Lotic
H-2
Cp P P
algae ^detritus ^sed ^particulate
(mg/kg) (mg/kg) (mg/kg) (mg/kg)
0.1
10.35
0.175
7.8
15.5
3
4.6
0.945
2.65
1.6
0.3
6.5
2.8
10.8
47.3
41
44
0.6
0.5
0.17
8.20
0.70
9.18
27.08
11.09
14.23
0.95
1.26
0.89
water
fag/L)
0.30
9.40
0.80
7.60
15.90
4.80
6.00
1.50
1.00
5.00
Draf
Site
EF
(L/g)
0.577
0.873
0.875
1.208
1.703
2.311
2.371
0.630
1.261
0.179
t Documi
-------
Reference
Butler et al.
1995
Butler et al.
1995
Butler et al.
1995
Butler et al.
1995
Butler et al.
1995
Butler et al.
1995
Butler et al.
1995
Butler et al.
1995
Butler et al.
1995
Butler et al.
1995
Butler et al.
1997
Butler et al.
1997
Butler et al.
1997
Butler et al.
1997
Butler et al.
1997
Site description
McElmo Cr.upstream from
Yellow Jacket Cyn.
McElmo Cr. downstream
from Yellow Jacket Cyn.
Hartman Draw near mouth,
at Cortez
Navajo Wash near Towaoc
San Juan River at Four
Comers
Cahone Canyon at
Highway 666
San Juan River at Mexican
Hat Utah
McElmo Cr. downstream
from Alkali Cyn.
Woods Cyn. Near Yellow
Jacket
McElmo Cr. at Hwy. 160,
near Cortez
Pond on Cahone Canyon,
west of 1 5 Road
West pond at CC Road
Large pond on Dove Creek
Pond on Woods Canyon at
15 Road
Large pond south of G
Road, southern Mancos
Valley
Site ID
ME3
ME4
HD2
NW
SJ1
CH
SJ3
ME2
we
ME1
CHP
PVP1
DCP1
WCP
MNP2
Specific
Specific waterbody
waterbody type -
type - Lentic or
original Lotic
Creek
Creek
Draw
Wash
River
Creek
River
Creek
Creek
Creek
Pond
Pond
Pond
Pond
Pond
Lotic
Lotic
Lotic
Lotic
Lotic
Lotic
Lotic
Lotic
Lotic
Lotic
Lentic
Lentic
Lentic
Lentic
Lentic
CP P P
algae ^detritus ^sed ^participate
(mg/kg) (mg/kg) (mg/kg) (mg/kg)
0.82
1.035
0.445
3.45
0.515
2.5
0.94
1.105
o o
J.J
1.8
4
1.5
1
2.3
5.4
0.4
0.5
0.2
1.6
0.3
4.3
0.2
1.1
1.5
2.1
1.4
2.1
3.2
6.7
0.57
0.72
0.30
2.35
0.39
3.28
0.43
1.10
2.22
1.80
2.90
1.45
1.45
2.71
6.01
water
(Hg/L)
6.00
6.00
2.00
12.00
1.50
12.00
1.50
3.00
5.50
2.00
5.00
2.00
2.00
3.00
3.00
Site
EF
(L/g)
0.095
0.120
0.149
0.196
0.262
0.273
0.289
0.367
0.405
0.900
0.580
0.725
0.725
0.904
2.005
Do not distribute, quote, or cite
H-3
Draft Document
-------
Reference
Butler et al.
1997
Butler et al.
1997
Butler et al.
1997
Butler et al.
1997
Butler et al.
1997
Butler et al.
1997
Butler et al.
1997
Casey 2005
Casey 2005
Formation
2012
Formation
2012
Formation
2012
Formation
2012
Formation
2012
Formation
2012
Formation
2012
Site description
Pond downstream from site
MNP2, southern Mancos
Valley
Mud Creek at Highway 32,
near Cortez
Cahone Canyon at
Highway 666
Tributary of Yellow Jacket
Canyon at Highway 666
Tributary of Cahone
Cany on at 13 Road
Unnamed tributary of
Cross Canyon upstream
from Alkali Canyon
Unnamed tributary of Cow
Canyon at 8 Road
Luscar Creek
Deerlick Creek
Hoopes Spring - HS
Sage Creek - LSV2C
Hoopes Spring - HS3
Sage Creek - LSV4
Crow Creek - 1A
Crow Creek - 3A
Crow Creek - CC150
Site ID
MNP3
MUD2
CHI
YJ1
CH2
CCTR
COW
HS
LSV-2C
HS-3
LSV-4
CC-1A
CC-3A
CC-150
Specific
waterbody
type-
original
Pond
Creek
Creek
Creek
Creek
Creek
Creek
Creek
Creek
Spring
Creek
Spring
Creek
Creek
Creek
Creek
Specific
waterbody
type-
Lentic or
Lotic
Lentic
Lotic
Lotic
Lotic
Lotic
Lotic
Lotic
Lotic
Lotic
Lotic
Lotic
Lotic
Lotic
Lotic
Lotic
Lotic
algae
(mg/kg)
4.5
1.3
2.05
1.85
1.75
1.75
1.45
5.5
12
8.09
12
9.56
3.64
3.1
1.2
Cc c
detritus ^sed ^particulate
(mg/kg) (mg/kg) (mg/kg)
5.9 5.15
1.30
2.05
1.85
1.75
1.75
1.45
3.2 2.4 3.48
1 0.2 0.45
2.3 5.25
4.6 6.10
7 9.17
3.6 5.87
1.2 2.09
0.83 1.60
0.63 0.87
water
1.00
18.50
10.50
7.00
5.50
4.50
3.50
10.70
0.20
20.95
13.80
17.05
8.45
2.45
2.20
0.80
Site
EF
(L/g)
5.153
0.070
0.195
0.264
0.318
0.389
0.414
0.325
2.236
0.244
0.447
0.536
0.694
0.799
0.806
1.041
Do not distribute, quote, or cite
H-4
Draft Document
-------
Reference
Formation
2012
Formation
2012
Formation
2012
Formation
2012
Grasso et al.
1995
Hamilton and
Buhl 2004
Lemly 1985
Lemly 1985
Lemly 1985
McDonald and
Strosher 1998
Presser and
Luoma 2009
Reidel and
Sanders
Saiki and
Lowe 1987
Saiki and
Lowe 1987
Saiki and
Lowe 1987
Saiki and
Lowe 1987
Saiki and
Lowe 1987
Site description Site ID
Crow Creek - CCS 50 CC-350
Crow Creek - CC75 CC-75
South Fork Tincup Cr. SFTC- 1
Deer Creek DC-600
Arapahoe Wetlands Pond 17
lower East Mill Creek LEMC
Belews Lake
High Rock Lake
Badin Lake
Michel Cr. at Highway 3 ER 75 1
UPCW
Upper Peters canyon (dry) dry
Delaware River
Kesterson Pond 2
Kesterson Pond 11
Kesterson Pond 8
Volta Pond 26
Volta Pond 7
Specific
Specific waterbody
waterbody type -
type - Lentic or
original Lotic
Creek
Creek
Creek
Creek
Pond
Creek
Lake
Lake
Lake
Creek
Wash
River
Pond
Pond
Pond
Pond
Pond
Lotic
Lotic
Lotic
Lotic
Lentic
Lotic
Lentic
Lentic
Lentic
Lotic
Lotic
Lotic
Lentic
Lentic
Lentic
Lentic
Lentic
algae
(mg/kg)
1.5
1.01
0.725
4.545
1.87
25.7
44.1
6.2
7.7
1.26
1.2
152.7
18.15
136.5
0.416
CP P
detritus ^sed ^particulate
(mg/kg) (mg/kg) (mg/kg)
0.7
0.54
0.31
1.4
0.4
38.9
8.27
1.8
2.07
2.32
0.6
1
44.65 34.82
47.95 8.56
92 6.045
1.01 0.2895
1.39 0.39
1.02
0.74
0.47
2.52
0.86
31.62
19.10
3.34
3.99
1.71
0.85
1.00
61.92
19.53
42.34
0.50
0.74
water
(Hg/L)
0.86
0.52
0.44
1.62
1.00
24.00
10.91
0.67
0.32
7.10
3.20
0.30
195.85
38.60
70.35
0.53
0.63
Site
EF
(L/g)
1.163
1.187
1.324
1.550
0.865
1.317
1.750
4.986
12.476
0.241
0.265
3.333
0.316
0.506
0.602
0.935
1.169
Do not distribute, quote, or cite
H-5
Draft Document
-------
Reference
Saiki and
Lowe 1987
Saiki and
Lowe 1987
Saiki et al.
1993
Saiki et al.
1993
Saiki et al.
1993
Saiki et al.
1993
Schuler et al.
1990
Schuler et al.
1990
Schuler et al.
1990
Stephens et al.
1988
Stephens et al.
1988
Site description
San Luis Drain
Volta Wasteway
San Joaquin R. above Hills
Ferry Road
Salt Slough at the San Luis
National Wildlife Refuge
San Joaquin R. at Durham
Ferry State Recereation
Area
Mud Slough at Gun Club
Road
Kesterson National
Wildlife Refuge
Kesterson National
Wildlife Refuge
Kesterson National
Wildlife Refuge
Marsh 4720
Drain J3
Site ID
SJR2
GT4
SJR3
GT5
Kesterson
Pond 7
Kesterson
Pond 2
Kesterson
Pond 11
*
*
Specific
waterbody
type-
original
Drain
Wasteway
River
Slough
River
Slough
Pond
Pond
Pond
Marsh
Drain
Specific
waterbody
type-
Lentic or
Lotic
Lotic
Lotic
Lotic
Lotic
Lotic
Lotic
Lentic
Lentic
Lentic
Lentic
Lotic
CP P P
algae ^detritus ^sed ^participate
(mg/kg) (mg/kg) (mg/kg) (mg/kg)
67
0.873
1.25
1.39
0.445
4.5
87.1
52.5
53.7
2.1
24
275 79.9 113.76
2.03 0.244 0.76
5 2.50
8.4 3.42
1.25 0.75
14.95 8.20
5.9 22.67
9.3 22.10
11.5 24.85
4.2 2.97
48 33.94
water
(Hg/L)
316.50
0.74
7.00
8.00
1.00
6.00
100.00
90.00
40.00
31.00
110.00
Site
EF
(L/g)
0.359
1.029
0.357
0.427
0.746
1.367
0.227
0.246
0.621
0.096
0.309
Do not distribute, quote, or cite
H-6
Draft Document
-------
APPENDIX I: OBSERVED VERSUS PREDICTED EGG-
OVARY CONCENTRATIONS
Do not distribute, quote or cite 1-1 Draft Document
-------
The following table includes data for 169 individual fish tissue selenium measurements from the 53 sites where EFs could be
calculated. Observed egg-ovary fish tissue measurements were compared to predicted egg-ovary fish tissue measurements calculated
using equation 19 of the main text, also shown here for convenience.
C,
= Crater X
X EF X CF
(Equation 19)
These data were used to generate the observed to predicted egg-ovary concentration figure 18 of the main text. When the measured
tissue type was either muscle or whole body, it was converted to egg-ovary using taxa specific conversion factors. The predicted and
measured concentrations are highly correlated (r = 0.81, t(i6?) = 17.91, P < 0.001 ).
Study
Birkner 1978
Birkner 1978
Birkner 1978
Birkner 1978
Birkner 1978
Birkner 1978
Birkner 1978
Birkner 1978
Birkner 1978
Birkner 1978
Birkner 1978
Butler etal. 1991
Butler etal. 1991
Butler etal. 1991
Butler etal. 1991
Site
East Allen Reservoir, Medicine Bow
WY
Galett Lake, Laramie WY
Larimer Highway 9 Pond, Fort Collins
CO
Meeboer Lake, Laramie WY
Miller's Lake, Wellington CO
Miller's Lake, Wellington CO
Sweltzer Lake, Delta CO
Sweltzer Lake, Delta CO
Twin Butter Reservoir, Laramie WY
Twin Butter Reservoir, Laramie WY
Twin Butter Reservoir, Laramie WY
Uncompahgre River at Colona
Uncompahgre River at Colona
Uncompahgre River at Colona
Uncompahgre River at Colona
Species
Iowa darter
Iowa darter
northern plains
killifish
northern plains
killifish
fathead minnow
Iowa darter
fathead minnow
northern plains
killifish
fathead minnow
Iowa darter
northern plains
killifish
bluehead sucker
brown trout
brown trout
flannelmouth
sucker
Site
Water
(ns/l)
4.8
0.8
15.9
0.3
6.0
6.0
9.4
9.4
7.6
7.6
7.6
1.5
1.5
1.5
1.5
EF
d/g)
2.31
0.88
1.70
0.58
2.37
2.37
0.87
0.87
1.21
1.21
1.21
0.63
0.63
0.63
0.63
rprpriCOnip
3.08
3.08
2.44
2.44
2.77
3.08
2.77
2.44
2.77
3.08
2.44
1.21
2.49
2.49
1.64
CF
1.45
1.45
1.63
1.63
2.00
1.45
2.00
1.63
2.00
1.45
1.63
1.82
1.45
1.45
1.41
Pred.
E/O
(mg/kg)
49.51
3.13
107.9
0.69
78.79
63.52
45.43
32.70
50.83
40.98
36.59
2.08
3.41
3.41
2.19
Obs.
E/O
(mg/kg)
52.68
3.05
93.83
12.59
21.97
33.38
157.8
52.15
68.90
60.81
37.76
3.27
4.77
5.06
2.40
Obs.
tissue
type
WB
WB
WB
WB
WB
WB
WB
WB
WB
WB
WB
WB
WB
WB
WB
Do not distribute, quote or cite
1-2
Draft Document
-------
Study
Butler etal. 1991
Butler etal. 1991
Butler etal. 1991
Butler etal. 1993
Butler etal. 1993
Butler etal. 1993
Butler etal. 1993
Butler etal. 1993
Butler etal. 1993
Butler etal. 1993
Butler etal. 1993
Butler etal. 1993
Butler etal. 1993
Butler etal. 1993
Butler etal. 1993
Butler etal. 1993
Butler etal. 1995
Butler etal. 1995
Butler etal. 1995
Butler etal. 1995
Butler etal. 1995
Butler etal. 1995
Site
Uncompahgre River at Colona
Uncompahgre River at Colona
Uncompahgre River at Colona
Navajo Reservoir, Piedra River Arm,
near La Boca
Navajo Reservoir, Piedra River Arm,
near La Boca
Navajo Reservoir, Piedra River Arm,
near La Boca
Navajo Reservoir, Piedra River Arm,
near La Boca
Navajo Reservoir, Piedra River Arm,
near La Boca
Navajo Reservoir, Piedra River Arm,
near La Boca
Navajo Reservoir, Piedra River Arm,
near La Boca
Spring Cr. at La Boca
Spring Cr. at La Boca
Spring Cr. at La Boca
Spring Cr. at La Boca
Spring Cr. at La Boca
Spring Cr. at La Boca
Hartman Draw near mouth, at Cortez
Hartman Draw near mouth, at Cortez
Hartman Draw near mouth, at Cortez
Hartman Draw near mouth, at Cortez
Hartman Draw near mouth, at Cortez
Hartman Draw near mouth, at Cortez
Species
mottled sculpin
mottled sculpin
rainbow trout
brown trout
bullhead
bullhead
channel catfish
common carp
common carp
common carp
bluehead sucker
brown trout
brown trout
fathead minnow
fathead minnow
speckled dace
fathead minnow
fathead minnow
flannelmouth
sucker
flannelmouth
sucker
flannelmouth
sucker
flannelmouth
sucker
Site
Water
(ws/1)
1.5
1.5
1.5
1.0
1.0
1.0
1.0
1.0
1.0
1.0
5.5
5.0
5.5
5.0
5.5
5.0
2.0
2.0
2.0
2.0
2.0
2.0
EF
d/s)
0.63
0.63
0.63
1.26
1.26
1.26
1.26
1.26
1.26
1.26
0.18
0.18
0.18
0.18
0.18
0.18
0.15
0.15
0.15
0.15
0.15
0.15
rprpriCOnip
2.65
2.65
2.44
2.49
1.68
1.68
1.35
1.70
1.70
1.70
1.21
2.49
2.49
2.77
2.77
2.78
2.77
2.77
1.64
1.64
1.64
1.64
CF
1.63
1.63
2.44
1.45
1.63
1.63
1.63
1.92
1.92
1.92
1.82
1.45
1.45
2.00
2.00
2.00
2.00
2.00
1.41
1.41
1.41
1.41
Pred.
E/O
(mg/kg)
4.09
4.09
5.63
4.55
3.47
3.47
2.78
4.12
4.12
4.12
2.17
3.23
3.55
4.95
5.45
4.97
1.65
1.65
0.69
0.69
0.69
0.69
Obs.
E/O
(mg/kg)
4.25
7.19
6.88
6.20
2.29
3.43
2.62
6.15
5.19
6.15
12.91
1.74
4.92
16.38
11.98
23.96
3.00
3.20
0.69
0.76
0.87
1.35
Obs.
tissue
type
WB
WB
WB
E-O
WB
WB
WB
WB
WB
WB
WB
WB
WB
WB
WB
WB
WB
WB
WB
WB
WB
WB
Do not distribute, quote or cite
1-3
Draft Document
-------
Study
Butler etal. 1995
Butler etal. 1995
Butler etal. 1995
Butler etal. 1995
Butler etal. 1995
Butler etal. 1995
Butler etal. 1995
Butler etal. 1995
Butler etal. 1995
Butler etal. 1995
Butler etal. 1995
Butler etal. 1995
Butler etal. 1995
Butler etal. 1995
Butler etal. 1995
Butler etal. 1995
Butler etal. 1995
Butler etal. 1995
Site
McElmo Cr. at Hwy. 160, near Cortez
McElmo Cr. at Hwy. 160, near Cortez
McElmo Cr. downstream from Alkali
Cyn.
McElmo Cr. downstream from Alkali
Cyn.
McElmo Cr. downstream from Alkali
Cyn.
McElmo Cr. downstream from Alkali
Cyn.
McElmo Cr. downstream from Alkali
Cyn.
McElmo Cr. downstream from Alkali
Cyn.
McElmo Cr. downstream from Alkali
Cyn.
McElmo Cr. downstream from Alkali
Cyn.
McElmo Cr. downstream from Yellow
Jacket Cyn.
McElmo Cr. downstream from Yellow
Jacket Cyn.
McElmo Cr. downstream from Yellow
Jacket Cyn.
McElmo Cr. downstream from Yellow
Jacket Cyn.
McElmo Cr. downstream from Yellow
Jacket Cyn.
McElmo Cr. downstream from Yellow
Jacket Cyn.
McElmo Cr. downstream from Yellow
Jacket Cyn.
McElmo Cr. downstream from Yellow
Species
fathead minnow
speckled dace
bluehead sucker
bluehead sucker
fathead minnow
flannelmouth
sucker
flannelmouth
sucker
flannelmouth
sucker
flannelmouth
sucker
speckled dace
common carp
common carp
common carp
fathead minnow
fathead minnow
flannelmouth
sucker
flannelmouth
sucker
flannelmouth
Site
Water
(ws/1)
2.0
2.0
3.0
3.0
3.0
3.0
3.0
3.0
3.0
3.0
6.0
6.0
6.0
6.0
6.0
6.0
6.0
6.0
EF
(i/p)
0.90
0.90
0.37
0.37
0.37
0.37
0.37
0.37
0.37
0.37
0.12
0.12
0.12
0.12
0.12
0.12
0.12
0.12
rprpriCOnip
2.77
2.78
1.21
1.21
2.77
1.64
1.64
1.64
1.64
2.78
1.70
1.70
1.70
2.77
2.77
1.64
1.64
1.64
CF
2.00
2.00
1.82
1.82
2.00
1.41
1.41
1.41
1.41
2.00
1.92
1.92
1.92
2.00
2.00
1.41
1.41
1.41
Pred.
E/O
(mg/kg)
9.97
10.01
2.43
2.43
6.11
2.56
2.56
2.56
2.56
6.13
2.35
2.35
2.35
3.98
3.98
1.67
1.67
1.67
Obs.
E/O
(mg/kg)
11.18
12.78
1.51
2.36
9.59
2.25
1.97
2.82
3.10
12.18
7.49
7.11
7.30
2.80
11.78
2.11
1.83
2.68
Obs.
tissue
type
WB
WB
WB
WB
WB
WB
WB
WB
WB
WB
WB
WB
WB
WB
WB
WB
WB
WB
Do not distribute, quote or cite
1-4
Draft Document
-------
Study
Butler etal. 1995
Butler etal. 1995
Butler etal. 1995
Butler etal. 1995
Butler etal. 1995
Butler etal. 1995
Butler etal. 1995
Butler etal. 1995
Butler etal. 1995
Butler etal. 1995
Butler etal. 1995
Butler etal. 1995
Butler etal. 1995
Butler etal. 1995
Butler etal. 1995
Butler etal. 1995
Site
Jacket Cyn.
McElmo Cr. downstream from Yellow
Jacket Cyn.
McElmo Cr. downstream from Yellow
Jacket Cyn.
McElmo Cr. downstream from Yellow
Jacket Cyn.
McElmo Cr .upstream from Yellow
Jacket Cyn.
McElmo Cr .upstream from Yellow
Jacket Cyn.
McElmo Cr .upstream from Yellow
Jacket Cyn.
McElmo Cr .upstream from Yellow
Jacket Cyn.
McElmo Cr .upstream from Yellow
Jacket Cyn.
McElmo Cr .upstream from Yellow
Jacket Cyn.
McElmo Cr .upstream from Yellow
Jacket Cyn.
McElmo Cr .upstream from Yellow
Jacket Cyn.
McElmo Cr .upstream from Yellow
Jacket Cyn.
McElmo Cr .upstream from Yellow
Jacket Cyn.
McElmo Cr .upstream from Yellow
Jacket Cyn.
McElmo Cr .upstream from Yellow
Jacket Cyn.
McElmo Cr .upstream from Yellow
Jacket Cyn.
Species
sucker
flannelmouth
sucker
flannelmouth
sucker
red shiner
bluehead sucker
bluehead sucker
bullhead
common carp
common carp
fathead minnow
fathead minnow
fathead minnow
flannelmouth
sucker
flannelmouth
sucker
flannelmouth
sucker
flannelmouth
sucker
flannelmouth
sucker
Site
Water
(ws/1)
6.0
6.0
6.0
6.0
6.0
6.0
6.0
6.0
6.0
6.0
6.0
6.0
6.0
6.0
6.0
6.0
EF
(i/p)
0.12
0.12
0.12
0.10
0.10
0.10
0.10
0.10
0.10
0.10
0.10
0.10
0.10
0.10
0.10
0.10
rprpriCOnip
1.64
1.64
2.53
1.21
1.21
1.68
1.70
1.70
2.77
2.77
2.77
1.64
1.64
1.64
1.64
1.64
CF
1.41
1.41
2.00
1.82
1.82
1.63
1.92
1.92
2.00
2.00
2.00
1.41
1.41
1.41
1.41
1.41
Pred.
E/O
(mg/kg)
1.67
1.67
3.64
1.26
1.26
1.57
1.87
1.87
3.17
3.17
3.17
1.33
1.33
1.33
1.33
1.33
Obs.
E/O
(mg/kg)
3.38
4.23
10.19
3.27
3.09
4.90
8.45
9.99
8.59
10.58
8.79
2.40
2.40
2.96
3.38
5.07
Obs.
tissue
type
WB
WB
WB
WB
WB
WB
WB
WB
WB
WB
WB
WB
WB
WB
WB
WB
Do not distribute, quote or cite
1-5
Draft Document
-------
Study
Butler etal. 1995
Butler etal. 1995
Butler etal. 1995
Butler etal. 1995
Butler etal. 1995
Butler etal. 1995
Butler etal. 1995
Butler etal. 1995
Butler etal. 1995
Butler etal. 1995
Butler etal. 1995
Butler etal. 1995
Butler etal. 1995
Butler etal. 1995
Butler etal. 1995
Butler etal. 1995
Butler etal. 1995
Butler etal. 1995
Butler etal. 1995
Butler etal. 1995
Butler etal. 1995
Butler etal. 1995
Site
McElmo Cr .upstream from Yellow
Jacket Cyn.
McElmo Cr .upstream from Yellow
Jacket Cyn.
McElmo Cr .upstream from Yellow
Jacket Cyn.
McElmo Cr .upstream from Yellow
Jacket Cyn.
McElmo Cr .upstream from Yellow
Jacket Cyn.
McElmo Cr .upstream from Yellow
Jacket Cyn.
Navajo Wash near Towaoc
Navajo Wash near Towaoc
Navajo Wash near Towaoc
San Juan River at Four Comers
San Juan River at Four Comers
San Juan River at Four Comers
San Juan River at Four Comers
San Juan River at Four Comers
San Juan River at Four Comers
San Juan River at Four Comers
San Juan River at Four Comers
San Juan River at Four Comers
San Juan River at Four Comers
San Juan River at Four Comers
San Juan River at Four Comers
San Juan River at Four Comers
Species
green sunfish
red shiner
red shiner
speckled dace
speckled dace
speckled dace
bluehead sucker
bluehead sucker
speckled dace
bluehead sucker
bluehead sucker
bluehead sucker
channel catfish
channel catfish
common carp
common carp
flannelmouth
sucker
flannelmouth
sucker
flannelmouth
sucker
flannelmouth
sucker
flannelmouth
sucker
flannelmouth
Site
Water
(ws/1)
6.0
6.0
6.0
6.0
6.0
6.0
12.0
12.0
12.0
1.5
1.5
1.5
1.5
1.5
1.5
1.5
1.5
1.5
1.5
1.5
1.5
1.5
EF
d/s)
0.10
0.10
0.10
0.10
0.10
0.10
0.20
0.20
0.20
0.26
0.26
0.26
0.26
0.26
0.26
0.26
0.26
0.26
0.26
0.26
0.26
0.26
rprpriCOnip
2.44
2.53
2.53
2.78
2.78
2.78
1.21
1.21
2.78
1.21
1.21
1.21
1.35
1.35
1.70
1.70
1.64
1.64
1.64
1.64
1.64
1.64
CF
1.45
2.00
2.00
2.00
2.00
2.00
1.82
1.82
2.00
1.82
1.82
1.82
1.63
1.63
1.92
1.92
1.41
1.41
1.41
1.41
1.41
1.41
Pred.
E/O
(mg/kg)
2.03
2.90
2.90
3.18
3.18
3.18
5.18
5.18
13.06
0.87
0.87
0.87
0.87
0.87
1.29
1.29
0.91
0.91
0.91
0.91
0.91
0.91
Obs.
E/O
(mg/kg)
7.26
9.19
8.39
5.59
13.98
10.98
16.91
13.09
17.37
2.18
1.71
2.18
2.85
6.70
10.18
6.53
2.70
2.11
3.10
0.86
1.55
5.92
Obs.
tissue
type
WB
WB
WB
WB
WB
WB
WB
WB
WB
WB
WB
WB
M
WB
WB
WB
M
WB
WB
WB
WB
WB
Do not distribute, quote or cite
1-6
Draft Document
-------
Study
Butler etal. 1995
Butler etal. 1995
Butler etal. 1995
Butler etal. 1995
Butler etal. 1995
Butler etal. 1995
Butler etal. 1995
Butler etal. 1995
Butler etal. 1995
Butler etal. 1995
Butler etal. 1995
Butler etal. 1995
Butler etal. 1995
Butler etal. 1995
Butler etal. 1995
Butler etal. 1995
Butler etal. 1995
Butler etal. 1995
Butler etal. 1997
Butler etal. 1997
Butler etal. 1997
Butler etal. 1997
Butler etal. 1997
Butler etal. 1997
Site
San Juan River at Four Comers
San Juan River at Four Comers
San Juan River at Four Comers
San Juan River at Four Comers
San Juan River at Mexican Hat Utah
San Juan River at Mexican Hat Utah
San Juan River at Mexican Hat Utah
San Juan River at Mexican Hat Utah
San Juan River at Mexican Hat Utah
San Juan River at Mexican Hat Utah
San Juan River at Mexican Hat Utah
San Juan River at Mexican Hat Utah
San Juan River at Mexican Hat Utah
San Juan River at Mexican Hat Utah
San Juan River at Mexican Hat Utah
Woods Cyn. Near Yellow Jacket
Woods Cyn. Near Yellow Jacket
Woods Cyn. Near Yellow Jacket
Cahone Canyon at Highway 666
Large pond south of G Road, southern
Mancos Valley
Mud Creek at Highway 32, near Cortez
Mud Creek at Highway 32, near Cortez
Mud Creek at Highway 32, near Cortez
Mud Creek at Highway 32, near Cortez
Species
sucker
red shiner
speckled dace
speckled dace
speckled dace
bluehead sucker
bluehead sucker
bluehead sucker
channel catfish
common carp
flannelmouth
sucker
flannelmouth
sucker
flannelmouth
sucker
flannelmouth
sucker
flannelmouth
sucker
flannelmouth
sucker
fathead minnow
fathead minnow
fathead minnow
green sunfish
fathead minnow
bluehead sucker
bluehead sucker
bluehead sucker
fathead minnow
Site
Water
(ws/1)
.5
.5
.5
.5
.5
.5
.5
.5
.5
1.5
1.5
1.5
1.5
1.5
1.5
5.5
5.5
5.5
10.5
3.0
18.5
18.5
18.5
18.5
EF
d/s)
0.26
0.26
0.26
0.26
0.29
0.29
0.29
0.29
0.29
0.29
0.29
0.29
0.29
0.29
0.29
0.40
0.40
0.40
0.20
2.00
0.07
0.07
0.07
0.07
rprpriCOnip
2.53
2.78
2.78
2.78
.21
.21
.21
.35
.70
1.64
1.64
1.64
1.64
1.64
1.64
2.77
2.77
2.77
2.44
2.77
1.21
1.21
1.21
2.77
CF
2.00
2.00
2.00
2.00
1.82
1.82
1.82
1.63
1.92
1.41
1.41
1.41
1.41
1.41
1.41
2.00
2.00
2.00
1.45
2.00
1.82
1.82
1.82
2.00
Pred.
E/O
(mg/kg)
1.99
2.19
2.19
2.19
0.96
0.96
0.96
0.96
1.42
1.00
1.00
1.00
1.00
1.00
1.00
12.32
12.32
12.32
7.27
33.31
2.87
2.87
2.87
7.20
Obs.
E/O
(mg/kg)
6.99
8.59
10.19
5.79
4.18
4.36
4.91
12.26
7.49
2.40
2.68
4.23
1.97
2.40
4.23
36.75
45.73
52.72
13.79
21.97
4.55
9.45
10.18
15.38
Obs.
tissue
type
WB
WB
WB
WB
WB
WB
WB
WB
WB
WB
WB
WB
WB
WB
WB
WB
WB
WB
WB
WB
WB
WB
WB
WB
Do not distribute, quote or cite
1-7
Draft Document
-------
Study
Butler etal. 1997
Butler etal. 1997
Butler etal. 1997
Butler etal. 1997
Butler etal. 1997
Butler etal. 1997
Butler etal. 1997
Casey 2005
Casey 2005
Casey 2005
Casey 2005
Grasso et al.
1995
Grasso et al.
1995
Grasso et al.
1995
Hamilton and
Buhl 2004
Lemly 1985
Lemly 1985
Lemly 1985
Lemly 1985
Lemly 1985
Lemly 1985
Lemly 1985
Lemly 1985
Lemly 1985
Lemly 1985
Lemly 1985
Lemly 1985
Site
Mud Creek at Highway 32, near Cortez
Mud Creek at Highway 32, near Cortez
Mud Creek at Highway 32, near Cortez
Mud Creek at Highway 32, near Cortez
Pond downstream from site MNP2,
southern Mancos Valley
Pond on Woods Canyon at 15 Road
Pond on Woods Canyon at 15 Road
Deerlick Creek
Deerlick Creek
Luscar Creek
Luscar Creek
Arapahoe Wetlands Pond
Arapahoe Wetlands Pond
Arapahoe Wetlands Pond
lower East Mill Creek
Badin Lake
Badin Lake
Badin Lake
Badin Lake
Badin Lake
Badin Lake
Belews Lake
Belews Lake
Belews Lake
Belews Lake
Belews Lake
Belews Lake
Species
fathead minnow
fathead minnow
green sunfish
green sunfish
smallmouth bass
fathead minnow
fathead minnow
rainbow trout
rainbow trout
rainbow trout
rainbow trout
fathead minnow
fathead minnow
fathead minnow
cutthroat trout
black bullhead
common carp
fathead minnow
green sunfish
mosquitofish
red shiner
black bullhead
common carp
fathead minnow
green sunfish
mosquitofish
red shiner
Site
Water
(ns/l)
18.5
18.5
18.5
18.5
1.0
3.0
3.0
0.2
0.2
10.7
10.7
1.0
1.0
1.0
24.0
0.3
0.3
0.3
0.3
0.3
0.3
10.9
10.9
10.9
10.9
10.9
10.9
EF
d/g)
0.07
0.07
0.07
0.07
5.15
0.90
0.90
2.24
2.24
0.33
0.33
0.86
0.86
0.86
1.32
12.48
12.48
12.48
12.48
12.48
12.48
1.75
1.75
1.75
1.75
1.75
1.75
rprpriCOnip
2.77
2.77
2.44
2.44
2.35
2.77
2.77
2.44
2.44
2.44
2.44
2.77
2.77
2.77
2.02
1.85
1.70
2.77
2.44
1.69
2.53
1.85
1.70
2.77
2.44
1.69
2.53
CF
2.00
2.00
1.45
1.45
1.42
2.00
2.00
2.44
2.44
2.44
2.44
2.00
2.00
2.00
1.96
1.63
1.92
2.00
1.45
1.63
2.00
1.63
1.92
2.00
1.45
1.63
2.00
Pred.
E/O
(mg/kg)
7.20
7.20
4.61
4.61
17.22
15.02
15.02
2.66
2.66
20.74
20.74
4.79
4.79
4.79
125.2
12.07
13.05
22.11
14.16
11.04
20.21
57.72
62.44
105.8
67.75
52.79
96.65
Obs.
E/O
(mg/kg)
23.96
12.98
11.03
10.16
17.03
19.97
29.96
3.14
8.16
16.79
33.48
13.16
13.18
14.58
102.7
4.26
5.81
3.17
3.25
5.53
4.45
28.62
38.97
28.75
20.84
44.94
38.59
Obs.
tissue
type
WB
WB
WB
WB
WB
WB
WB
M
E-O
M
E-O
WB
WB
WB
WB
M
M
M
M
M
M
M
M
M
M
M
M
Do not distribute, quote or cite
1-8
Draft Document
-------
Study
Lemly 1985
Lemly 1985
Lemly 1985
Lemly 1985
Lemly 1985
Lemly 1985
Saikietal. 1993
Saikietal. 1993
Saikietal. 1993
Saikietal. 1993
Saikietal. 1993
Saikietal. 1993
Saikietal. 1993
Saikietal. 1993
Saikietal. 1993
Saikietal. 1993
Saikietal. 1993
Saikietal. 1993
Saikietal. 1993
Saikietal. 1993
Saikietal. 1993
Saikietal. 1993
Stephens et al.
1988
Site
High Rock Lake
High Rock Lake
High Rock Lake
High Rock Lake
High Rock Lake
High Rock Lake
Mud Slough at Gun Club Road
Mud Slough at Gun Club Road
Mud Slough at Gun Club Road
Mud Slough at Gun Club Road
Salt Slough at the San Luis National
Wildlife Refuge
Salt Slough at the San Luis National
Wildlife Refuge
Salt Slough at the San Luis National
Wildlife Refuge
Salt Slough at the San Luis National
Wildlife Refuge
San Joaquin R. above Hills Ferry Road
San Joaquin R. above Hills Ferry Road
San Joaquin R. above Hills Ferry Road
San Joaquin R. above Hills Ferry Road
San Joaquin R. at Durham Ferry State
Recereation Area
San Joaquin R. at Durham Ferry State
Recereation Area
San Joaquin R. at Durham Ferry State
Recereation Area
San Joaquin R. at Durham Ferry State
Recereation Area
Marsh 4720
Species
black bullhead
common carp
fathead minnow
green sunfish
mosquitofish
red shiner
bluegill
bluegill
largemouth bass
largemouth bass
bluegill
bluegill
largemouth bass
largemouth bass
bluegill
bluegill
largemouth bass
largemouth bass
bluegill
bluegill
largemouth bass
largemouth bass
black bullhead
Site
Water
(ws/1)
0.7
0.7
0.7
0.7
0.7
0.7
6.0
6.0
6.0
6.0
8.0
8.0
8.0
8.0
7.0
7.0
7.0
7.0
1.0
1.0
1.0
1.0
31.0
EF
d/s)
4.99
4.99
4.99
4.99
4.99
4.99
1.37
1.37
1.37
1.37
0.43
0.43
0.43
0.43
0.36
0.36
0.36
0.36
0.75
0.75
0.75
0.75
0.10
rprpriCOnip
1.85
1.70
2.77
2.44
1.69
2.53
2.12
2.12
1.54
1.54
2.12
2.12
1.54
1.54
2.12
2.12
1.54
1.54
2.12
2.12
1.54
1.54
1.85
CF
1.63
1.92
2.00
1.45
1.63
2.00
2.13
2.13
1.42
1.42
2.13
2.13
1.42
1.42
2.13
2.13
1.42
1.42
2.13
2.13
1.42
1.42
1.63
Pred.
E/O
(mg/kg)
10.10
10.92
18.50
11.85
9.23
16.91
37.11
37.11
17.88
17.88
15.46
15.46
7.45
7.45
11.31
11.31
5.45
5.45
3.37
3.37
1.63
1.63
8.98
Obs.
E/O
(mg/kg)
5.35
4.49
4.00
3.13
5.85
4.62
13.65
10.67
9.65
9.79
9.60
9.17
6.67
5.68
7.04
5.76
3.12
3.41
4.27
4.05
2.55
2.41
11.44
Obs.
tissue
type
M
M
M
M
M
M
WB
WB
WB
WB
WB
WB
WB
WB
WB
WB
WB
WB
WB
WB
WB
WB
WB
Do not distribute, quote or cite
1-9
Draft Document
-------
Study
Stephens et al.
1988
Stephens et al.
1988
Site
Marsh 4720
Marsh 4720
Species
common carp
common carp
Site
Water
(ns/l)
31.0
31.0
EF
d/g)
0.10
0.10
rprpriCOnip
1.70
1.70
CF
1.92
1.92
Pred.
E/O
(mg/kg)
9.71
9.71
Obs.
E/O
(mg/kg)
36.49
40.33
Obs.
tissue
type
WB
WB
Do not distribute, quote or cite
1-10
Draft Document
-------
APPENDIX J: SUPPLEMENTARY INFORMATION ON
SELENIUM BIOACCUMULATION IN AQUATIC ANIMALS
Do not distribute, quote, or cite J-l Draft Document
-------
1.0 EFFECTS OF GROWTH RATE ON THE ACCUMULATION OF
SELENIUM IN FISH
EPA analyzed the effect of the growth rate parameter g when estimating selenium bioaccumulation using
the mechanistic bioaccumulation modeling described in Equation 1 of the main text. Because the addition
of tissue associated with growth could have a dilution effect on the chemicals present in tissue, a
parameter representing growth rate is present in the denominator of Equation 1. Indeed, growth can be an
important factor in the bioaccumulation of very hydrophobic chemicals with low excretion rates such as
polychlorinated biphenyls, (Connolly and Pedersen 1988). However, the effect of growth may not be as
important for selenium because of its unique biogeochemical characteristics, route of exposure, and role
as a micronutrient.
EPA tested the effect of the growth rate parameter g on estimates of selenium bioaccumulation using
Equation 1 with different food web scenarios. Increasing growth rates from 0 (no growth) to 0.2/day (a
relatively high rate of growth) reduced selenium concentrations in trophic level 2 and 3 organisms by as
much as a factor of 10 to 20. Thus incorporating growth rate in Equation 1 could result in significant
dilution of selenium and lower estimates of selenium bioaccumulation.
Although increasing the value of the growth parameter g in Equation 1 reduces estimates of selenium
bioaccumulation, this simple analysis neglects an important physiological linkage between growth and
food consumption. Organisms must consume enough food to support growth and meet their energy
requirements for respiration, specific dynamic action, waste loss, and reproduction. These physiological
requirements suggest that higher growth rates are associated with greater rates of food consumption.
Because food consumption is the primary route of selenium exposure in aquatic organisms, increased
selenium exposure associated with higher food consumption could counterbalance the dilution of
selenium in tissue associated with higher growth rates.
EPA tested the effects of growth on estimates of selenium bioaccumulation using Equation 1 when
increased food consumption was associated with higher growth rates. EPA modified Equation 1 to
incorporate a simple relationship for bioenergetics (Thomann et al. 1992) and applied the model to
reexamine the sensitivity of selenium bioaccumulation to growth rates in trophic level 2 and 3 organisms.
The results of this analysis showed that increasing growth rates over two orders of magnitude increased
selenium concentrations in trophic level 2 by a factor of 2, and decreased selenium concentrations in
trophic level 3 by 10%. When growth rates were increased simultaneously in trophic levels 2 and 3, the
selenium concentrations increased by less than a factor of 2. This analysis suggests that when
Do not distribute, quote, or cite J-2 Draft Document
-------
bioenergetics is considered, selenium bioaccumulation is generally insensitive to organism growth rates.
EPA believes that uncertainties in the toxicokinetic parameters of selenium far outweigh the effects on
growth rate on selenium bioaccumulation. Thus, the growth rate parameter g was removed from Equation
1 for the purpose of deriving a translation equation that could be used to implement a tissue-based
selenium water quality criterion.
2.0 ANALYSIS OF THE RELATIVE CONTRIBUTION OF AQUEOUS AND
DIETARY UPTAKE ON THE BIOACCUMULATION OF SELENIUM
EPA analyzed the relative contributions of direct aqueous uptake versus ingestion of selenium in
consideration of removing the uptake rate constant ku from Equation 1. Because an important exposure
route for some chemicals is direct contact with water, an uptake rate constant ku is present in the
numerator of Equation 1. However, fish and invertebrate organisms absorb selenium primarily through
the consumption of food rather than from direct aqueous uptake (Forester 2007; Lemly 1985; Luoma et
al. 1992). Thus, removing the uptake rate constant ku could simplify Equation 1 while maintaining the key
determinants of selenium bioaccumulation.
EPA tested the relative contribution of aqueous versus dietary uptake of selenium using a version of
Equation 1 that incorporates both exposure pathways (Thomann et. al. 1992). For trophic level 2,
selenium bioaccumulation was estimated for a range of uptake rates that varied according to the
respiration rate and aqueous transfer efficiency of selenium relative to dissolved oxygen. For trophic level
3, uptake rates were varied within a range of values reported in Besser et al. (1993) and Bertram and
Brooks (1986).
EPA's analysis showed that diet accounted for 34% - 92% of selenium bioaccumulation at trophic level 2,
with a median of 74%. At trophic level 3, diet accounted for 62% -100% of tissue selenium, with a
median of 95%. Thus, disregarding aqueous uptake of selenium only resulted in a small (-5%) reduction
in estimated selenium bioaccumulation in trophic level 3 organisms. These results are consistent with
previous studies indicating that diet is the primary exposure route of selenium, and suggests that the
uptake rate constant for selenium can be removed from Equation 1 with negligible effect for higher
trophic levels organisms.
Do not distribute, quote, or cite J-3 Draft Document
-------
3.0 KINETICS OF ACCUMULATION AND DEPURATION: AVERAGING
PERIOD
3.1 Background
For setting averaging periods for aquatic life criteria, U.S. EPA (1995b) used the concept that the criterion
averaging period should be less than or equal to the "characteristic time" describing the toxic speed of
action. In the context of the water-borne direct toxicity of metals, characteristic time = 1/k, where k is the
first-order kinetic coefficient in a toxico-kinetic model fitted to the relationship between LC50 and
exposure duration.
In the context of selenium bioaccumulation in a single trophic level, k would the first-order depuration
coefficient, and 1/k would equal the time needed to depurate to a concentration of 1/e times the initial
concentration (where e=2.718). For depuration of multiple trophic levels sequentially, the characteristic
time is likewise the time needed for c/c0 to reach a value of 1/e, as shown in Figure Jla. The accumulation
curve is the inverted depuration curve, as shown in Figure Jib.
0.8 fc-V--
(C
4-1
10 0 6
o
4-*
_>
JS 0.4
u
XX^i i
-1-- -L/e ,• .^ i :
L** — — — TL1 on fixed water cone
.1 y
.• | f — — , TL2 on accum TLl
^^^— TL3 on accum TL2
ill
20 40 60 80 100
Time, days
Figures Jl a & b. Depuration and accumulation behavior for algae-detritus-sediment k=0.2/day,
invertebrate k=0.2/day and fish k=0.02/day, calculated with time step = 0.1 day. Concentration is
expressed as a dimensionless ratio: concentration at time t divided by either starting concentration
(Jla) or plateau concentration (Jib).
In the Figures Jl a & b examples, the characteristic time for algae-detritus-sediment is 5 days, the
characteristic time for invertebrates on an invariant diet is 5 days, the characteristic time for fish on an
invariant diet is 50 days, and the characteristic time for fish on an invertebrate diet that is itself depurating
or accumulating is the approximate sum of the individual characteristic times, or ~60 days.
Do not distribute, quote, or cite
J-4
Draft Document
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In contrast to the model depuration rate, k, the model uptake rate (AE, assimilation efficiency, multiplied
by IR, intake rate) does not affect the characteristic response time. Rather it affects the magnitude of the
accumulation plateau. Uptake rate thus affects the TTF value itself but is not relevant to setting an
averaging period.
Because short averaging periods are more environmentally conservative than long averaging periods,
selecting parameter values for fast kinetics is more environmentally conservative. Figure Jl reflects
environmentally conservative choices for k values.
3.2 Approach for Modeling Effects of Time- Variable Se Concentrations
Expression of concentrations. None of the concentrations in this analysis are expressed in ordinary units
of concentration. All concentrations are modeled as values normalized to their allowable benchmark
concentration - that is, concentration = 1 for a particular medium (water, algae-detritus-sediment,
invertebrates, or fish) means that the medium is at its criterion concentration or corresponding benchmark.
It is assumed that the benchmarks correctly align - water held at its benchmark concentration will
ultimately yield Trophic Levels 1, 2, and 3 at their respective benchmark concentrations. The Trophic
Level 3 benchmark is the reproductive EC10 for the 5th percentile taxon: i.e., the fish tissue criterion.
Formulation of the bioaccumulation model for kinetic analysis. For algae-detritus-sediment, for
invertebrates, and for fish, accumulation at time t equals accumulation at time t-1 plus intake minus
depuration, as follows:
Algae-detritus-sediment:
CTLi[t] = CTLl[t-l] + kuptake C[t-l]water-kTLl CTLl[t-l]
Invertebrates:
CTL2[t] = CTL2[t-l] + AETL2IRTL2 CTL1[t-l] - kTL2 CTL2[t-l]
Fish:
CTL3[t] = CTL3[t-l] + AETL3IRTL3 CTL2[t-l] - kTL3 CTL3[t-l]
For algae-detritus-sediment, the depuration rate k is assigned a value of 0.2/day, similar to the sum of
depuration and growth-dilution rate coefficients used by Brix and DeForest (2008). Because a lentic
system would involve the slower kinetics of sediment exchange, the rapid rate used here implies a lotic
system.
Do not distribute, quote, or cite J-5 Draft Document
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For invertebrates, a value of 0.2/day was assigned, considerably higher than those for Lumbriculus, Asian
clam, zebra mussel, but close to those of mayfly and copepods, which are very small in size. As
previously mentioned, higher k (more rapid kinetics) is an environmentally conservative assumption in
this context.
For fish, the median depuration coefficient measured by Bertram and Brooks (1986) for 6-9 month-old
(early adult) fathead minnows was used, providing a kTL3 value of 0.02/day. Because of the small size of
adults of this species, this represents faster kinetics than would likely be applicable the salmonids and
centrarchids of greatest concern for selenium toxicity. The striped bass k value of Baines et al. (2002) is
inapplicable here because it was measured in the early juvenile life stage, a size that is too small to be
relevant to reproductive impairment stemming from exposure of adult females. The concentration in fish
could be equivalently viewed as either whole body or egg-ovary, relative to their respective benchmarks.
That is, partitioning within body of the fish is assumed not to involve a time delay.
The value of a TTF is given by AE x IR/k (or kuptake/k for algae-detritus-sediment). Concentrations in
TL1, TL2, and TL3 are normalized to their benchmarks, meaning that all benchmark concentrations have
a value of 1.0. In this normalized context, the TTFs must also equal 1.0, since upon reaching steady state,
TL1 at its benchmark will yield TL2 at its benchmark, which in turn will yield TL3 at its benchmark.
Again, the analysis is not intended to reflect actual concentrations, merely portray temporal behavior.
Since 1 = TTF = AE x IR/k, it follows that AE x IR = k within this normalized framework. Although only
the product AE x IR is relevant, they are retained as distinct parameters to maintain parallelism with
remainder of the criterion document. AE was assigned a value of 0.5 for fish and invertebrates, and IR =
k/AE in the normalized framework.
Time step durations of 0.1-1.0 day were considered. Short time steps increase accuracy by decreasing the
numerical dispersion inherent in expressing C[t] = f(C[t-l]). A time step of 0.5 day was found to yield
sufficient accuracy, as measured by predicted values at the characteristic time for depuration or
accumulation (per Figure J-l).
Prediction of Effects. The effect level associated with the tissue concentration at any time t is calculated
via the log probit concentration-response curve, one of the commonly used sigmoid curves. It assumes
that the sensitivities in the underlying population are log-normally distributed such that the concentration
yielding effects on k percentage of the population is given by:
Do not distribute, quote, or cite J-6 Draft Document
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ECk = EC50 exp(o z)
where o is the inverse of the concentration-response curve slope and z is the normal deviate
corresponding to k percent (e.g., for k=10%, z=NORMSINV(0. !)=-!.28155). Among the reproductive
impairment studies presented in Appendix C, an approximate median ratio for EC50/EC10 is 1.5. This
translates to o=0.3164.
Since the fish tissue criterion concentration equals 1.0 in this normalized framework, at any time t, the
fractional level of effect corresponding to any value of CTLS is given by:
Fractional Effect [t] = NORMSDIST(z[t])
where z[t] is given by:
zftj = LN(CTL3[t]/l. 5)/0.3164
Exposure Scenarios. Three exposure scenarios were evaluated under which the water criterion was just
barely attained. The first two are absolute worst case scenarios, in which the 30-day average water
concentration remains continuously at the criterion concentration at all times. The third is a realistic
scemario.
1. Steady concentrations at the criterion: this is worst-case continuous exposure. In the real world
this could not occur because water concentrations vary substantially over time. For the 30-day
average concentration not to exceed more than once in three years, the realistically varying daily
concentrations must remain well below the criterion concentration a large majority of the time.
2. Uniform 1-day spikes at 3OX the water criterion concentration, occurring at uniform 30-day
intervals (i.e., separated by 29 days of zero concentration) such that the 30-day average always
equals the criterion. This is the worst-case intermittent scenario, attaining the criterion through a
time series that continually maximizes the 30-day average exposure at the water criterion
concentration while also imposing the highest variability possible from spikes of 1-day duration.
In the real world intermittent runoff sources do not occur at uniform intervals: merely averaging
30-days between discharges would yield an exceedance each time the discharge occurred with
Do not distribute, quote, or cite J-7 Draft Document
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less than 30-days spacing. Further, the once-per-month peak concentrations could never be
controlled at exactly 3 OX the chronic water criterion per the above discussion of the first scenario.
It is because they lack real-world random variability that the above two scenarios are not realistic.
They are used as absolute worst cases for purposes of comparison. The following third scenario
represents a realistic and indeed typical situation for continuous exposure:
3. Log-normally distributed, smoothly variable concentrations with the 30-day average exceeding
the criterion once in three years when counted using the procedure of EPA (1986). The log
standard deviation of 0.5 applied here represents typical real-world time variability for
continuously flowing waters. The log serial correlation coefficient p = 0.8 represents that typical
of smaller streams.
With respect to maximizing toxic effects while attaining the criterion, Scenarios #1 and #2 are absolute
worst cases. In contrast, Scenario #3 represents typical time variability in ambient waters. This third
scenario requires randomly generated concentrations (having specified target statistical characteristics).
Multiple runs of long series are therefore needed to assure some reasonable degree of accuracy. A
minimum of 20 runs of random series of 3000 days were used. The concentrations at each half-day time
step were generated by the following formula:
Cftjwater = C[t-l]water^(p') * GM*(l-p') * EXP{a * SQRT(l-p 'A2j *NORMSINV(RAND)}
where p' (rho prime) is the desired serial correlation coefficient between half-day time steps: p'=SQRT(p)
[approximation], where p (rho) is the desired serial correlation coefficient between daily values; GM is
the desired geometric mean or median, and o is the desired log standard deviation. The above formula
allows a time series with the desired statistical characteristics to be generated.
3.2.1 Model Results
3.2.1.1 Steady concentrations at the water criterion concentration.
No graphic is needed to explain this scenario. With water steady at its criterion, algae-detritus-sediment
and invertebrates are likewise steady at their benchmark concentrations, and fish tissue is at its criterion
concentration. For the 5th percentile taxon, the effect would thus be 10% since the concentration is steady
attheEClO.
Do not distribute, quote, or cite J-8 Draft Document
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3.2.1.2 Uniformly spaced spikes at maximum concentrations
Figure J2. Scenario 2, uniform 1-day spikes at SOX the water criterion concentration, occurring at uniform
30-day intervals such that the 30-day average always equals the criterion. Read invertebrate and fish
tissue concentrations on left scale, water concentrations on right scale. Time=0 does not represent the
beginning of exposure; prior to Time=0 the same exposure pattern had been going on for a long time
(e.g., 10,000 days).
7
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-------
The key point here is that attaining the 30-day average via 1-day spikes spaced 30 days apart generates a
small oscillation in fish tissue concentrations. Averaged over the 30-days, the fish tissue concentrations
exactly attain their criterion and the predicted effect is 10%.
Do not distribute, quote, or cite J-10 Draft Document
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3.2.1.3 Log-normally distributed, smoothly varying concentrations
This is the most realistic scenarios, corresponding to typical variability observed in streams.
2.5 i [[[
I - fish tissue
c |
2 | water daily
• ...... water 30-davg
™ 2
U
o
+*
B 1.5
0 500 1000 1500 2000 2500 3000
Time, days
Figure J3. A typical example of log-normally distributed, smoothly variable concentrations. The
standard deviation of natural logs is 0.5 and the serial correlation coefficient of logs is 0.8 for daily
values, both typical real-world situations. (The compression of 3000 days into the graph might make it
difficult to recognize that the time series is smoothly varying - it has serial correlation.) At time=0, TL1,
TL2, and TL3 begin at their average concentrations.
In the Figure J3 example run, instantaneous water concentrations exceed the 30-day average criterion 7%
of the time. The 30-day average concentrations exceed the criterion 1.05 times per 3 year period, counted
per the EPA (1986) counting method. Tissue concentrations do not exceed their criterion at any time, and
-------
3.2.2 Summary of Scenario Results
Because Scenario 3 involves generation of random concentrations, the above graphs show just one run
(3000 days) for each. Full results for the 20 runs of that scenario is shown below.
Scenario
1. Steady
2. Uniform
spikes
3. Smooth
variable
Water:
# 30-day avg.
exceedances /
3-yr1
0.00
0.00
1.01
Water:
% of time
exceeding
0.00
3.33
7.8
Tissue:
% of time
exceeding
0.00
56.7
0.00
Mean
effect
for 5th
%ile
Taxon
10.0
10.0
0.18
Comment
Steady at water and tissue
benchmarks
30-d avg water cone, remains
steady at benchmark (Fig. J2)
Median=0.49 x benchmark, log
stdev=0.5, rho(daily)=0.8 (e.g.,
Fig. 5) 2
1. Counting procedure for 30-d avg. exceedances is that of U.S. EPA (1986).
2. Results for Scenario 3 are average of 20 runs of 3000 days, each run with 0.6-1.4 exceedances / 3 yr.
Runs not yielding exceedances within these bounds were not used. Among the 20 runs used, the effect
CV=0.35.
It can be concluded that the kinetics of selenium accumulation and depuration are sufficiently slow that
applying a 30-day averaging period to the water criterion concentration affords protection even under
unrealistic worst case conditions.
3.2.3 Example Responses to Increases in Water Concentrations
The previous Figures J2 and J3 illustrate situations after achievement of a dynamic steady state, where
daily water concentrations change but longer-term mean water concentrations do not change. Given the
same kinetic parameters as used above (i.e., yielding a 60-day characteristic time), this section addresses
the rate at which tissue concentrations respond to increases in mean water concentrations, for example as
would result from a new source. This is similar to the rising curve previously shown in Figure Jib. The
rapid kinetics used here for the water-TLl step imply a small lotic system having little involvement of the
bed sediments.
3.2.3.1 Step-Function Example
This example addresses the question: If water concentrations are increased to a level that is slightly too
high, ultimately (at Time=oo) yielding fish-tissue concentrations at the EC20 instead of the EC 10, how
long would it take for those tissue concentrations to rise to a level that exceeds the (EClO-based)
criterion?
Do not distribute, quote, or cite
J-12
Draft Document
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Prior to Time=0 in this example the concentrations in TL3 had been at a moderate background
concentration of 0.406 times the criterion, corresponding to the median West Virginia reference-site egg
concentrations tabulated by West Virginia Department of Environmental Protection (2010). The
concentrations in TL1 and TL2 are likewise assumed to have been at 0.406 normalized to their
corresponding benchmarks. At Time=0 the water concentrations increase such that ultimately they will
produce an effect 10% higher than the target, thus at the EC20 of the hypothetical 5th percentile sensitive
species. For typical selenium concentration-response slopes, this is 1.15-fold above the EC 10. Figure J4
illustrates this scenario, which shows that 90 days are needed for TL3 concentrations to rise above the
criterion.
2.5
.O
*k
01
_*J
*b
u
o
** -i r
re 1.5
O
U
re
•fish tissue
water daily
water 30-d avg
0.5
tfl
tf)
50 100 150 200 250
Time, days
300
350
400
Figure J4. TL3 concentration responding to a Time=0 step-function increase in water concentration
that remains time-invariant thereafter. Given that the water concentration is too high, ultimately
yielding tissue concentrations at the hypothetical sensitive species EC20, 1.15-fold above the criterion,
and given the previously presented kinetic parameters, it is calculated to take 90 days for TL3
concentrations to rise above the criterion.
Do not distribute, quote, or cite
J-13
Draft Document
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3.2.3.2 Continuously Time-Variable Example for Flowing Waters
To provide more realism, this example considers typical time variability, following up on Figure J3. In
this example, prior to Time=0, TL1, TL2, and TL3 concentrations were at a low background
concentration, 0.1 normalized to their criterion or respective benchmark. At Time=0 begin water
concentrations having median = geometric mean = 0.49 normalized as a dimensionless ratio,
concentration/criterion. Because the water concentrations are log-normally distributed, with log standard
deviation = 0.5, the arithmetic mean is higher than the median and has the normalized value 0.56. If the
simulation went on for a very long time, this time series (designed to have geometric mean 0.49 times the
criterion, log standard deviation 0.5, and log serial correlation coefficient 0.8) would average one
exceedance every three years, when exceedances are counted using the EPA (1986) approach. Figure J5
shows atypical short series of 400 days.
2.5
Q>
_*j
~
U
O
JD 1.5
CH
u
E
O
U 1
(D
•fish tissue
water daily
water 30-d avg
50 100 150 200 250
Time, days
300
350
400
Figure J5. Flowing water example of TL3 concentration starting at a concentration of 0.1
normalized to the criterion, and responding to randomly varying log-normally distributed water
concentrations having median 0.49 (expressed as a dimensionless ratio: concentration/criterion), log
standard deviation 0.5, and log serial correlation coefficient 0.8. Again, all concentrations are as
dimensionless ratios relative to the criteria concentrations.
Do not distribute, quote, or cite
J-14
Draft Document
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Several points are worth noting. Because the water concentrations happen (by chance) to be below
average for the first 50 days, the TL3 concentrations rise somewhat slowly during that period. Were they
to be above average during that period, the TL3 concentrations would more rapidly approach their
dynamically varying plateau. In such a short time series it is not graphically apparent what the long-term
average TL3 concentration will be; however, because the long-term arithmetic mean water concentration
would be 0.56 (normalized the its criterion), the TL3 concentration would likewise end up averaging 0.56
normalized to its criterion, if tracked for many years.
It is also worth noting that most 400-day series of the type shown in Figure J5 would not have
occurrences of 30-day average concentrations above the criterion (as suggested by Figure J3). This
particular random series does have a period of 30-day average exceedances, near Day 300, but it does not
persist long enough to cause the TL3 concentration to approach its criterion.
Lastly, it should be noted that when concentrations are randomly varying as in Figure J5, the water
concentrations that one observes are highly dependent on when the samples are taken. The TL3
concentrations observed are far less dependent on when the samples are taken (after the plateau is
approached), but time variations, although muted, are still present.
The example scenarios depicted here show lotic time to steady state of approximately 3 months to less
than 1 year under different discharge scenarios including both continuous and intermittent discharges.
The scenarios also assume that the new selenium input is from one source; multiple new sources
particularly with varying discharge patterns, might have a different response tme and pattern for various
trophic levels.
The example is likely not appropriate for lentic systems, because they would not be expected to have the
rapidly varying water concentrations of Figure J5. In addition, the water-to-TLl kinetics would likely be
slower in lentic systems with new or time-varying sources because of the role of bottom sediments acting
as a reservoir in recycling selenium. Ultimately this should yield slower rising and smoother TL3
concentrations compared to those in Figure J5.
Do not distribute, quote, or cite J-15 Draft Document
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APPENDIX K: SITE-SPECIFIC CRITERIA
Do not distribute, quote, or cite K-l Draft Document
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1.0 TRANSLATING THE CONCENTRATION OF SELENIUM IN TISSUE TO
A CONCENTRATION IN WATER
The EPA is recommending a selenium criterion to protect aquatic life that is expressed as a concentration
in the eggs or ovaries offish. Although the selenium concentration in eggs or ovaries is the most sensitive
and reliable basis for a criterion, implementation can be challenging because most state and tribal Clean
Water Act programs require the expression of water quality criteria as an ambient concentration in the
water-column. Therefore, the EPA is also recommending two water-column criterion values, one for lotic
or flowing waters, and the other for lentic or still waters.
The EPA derived the water-column criterion values by developing a translation equation based on
selenium bioaccumulation modeling. The EPA worked with the United States Geological Survey to
derive a translation equation that utilizes a mechanistic model of bioaccumulation previously published in
peer-reviewed scientific literature (Luoma et. al., 1992; Wang et. al., 1996; Luoma and Fisher, 1997;
Wang, 2001; Schlekat et al. 2002b; Luoma and Rainbow 2005; Presser and Luoma 2006; Presser and
Luoma 2010; Presser 2013). The selenium egg-ovary FCV is translated to water concentration values at a
set of lentic and lotic aquatic systems, and the distribution of site-specific water concentrations from these
sites is used to derive water-column criterion values protective of aquatic life. This appendix describes
how states and tribes may use this methodology to translate the egg-ovary FCV into site-specific water-
column concentrations to more precisely manage selenium in specific aquatic systems. The use of a
Bioaccumulation Factor (BAF) approach is also briefly discussed. States and tribes may also derive site-
specific water concentration values using other scientifically defensible methods.
The relationship between the concentration of selenium in the eggs or ovaries of fish and the
concentration of selenium in the water-column can vary from one aquatic system to another. The species
offish, the species and proportion of prey, and a variety of site-specific biogeochemical factors can
substantially affect selenium bioaccumulation and thus determine the allowable concentration of selenium
in surface waters that is protective of aquatic life. Because most state and tribal Clean Water Act
programs require the expression of water quality criteria as ambient concentrations in water,
implementation of the selenium criterion expressed as a concentration in the eggs or ovaries offish
requires the ability to translate the egg-ovary FCV into site-specific water-column concentrations. The
EPA considered two different modeling approaches to implement the selenium egg-ovary FCV:
mechanistic model of bioaccumulation, and a site-specific, field-derived bioaccumulation factor (BAF).
Do not distribute, quote, or cite K-2 Draft Document
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The mechanistic modeling approach uses scientific knowledge of the physical and chemical processes
underlying bioaccumulation to establish a relationship between the concentrations of a chemical in the
water-column and the concentration of a metal in the tissue of aquatic organisms. The EPA used this
approach to develop a mathematical equation that allows states and tribes to formulate site-specific
models of trophic transfer of selenium through aquatic food webs and translate the egg-ovary FCV into an
equivalent site-specific water concentration.
The BAF approach is an empirical model of bioaccumulation based on the measured concentration of a
chemical found in the tissue of aquatic organisms and the water from where the aquatic organisms reside
(U.S. EPA 2001c). The concentration of the chemical is measured in both fish tissue and the water-
column, and a BAF is calculated by taking the ratio of the two concentrations. The BAF can then be used
to estimate the concentration of the chemical in one media when only the concentration of the chemical in
the other media is known.
Both the mechanistic modeling approach and the BAF approach have advantages and disadvantages that
states and tribes should consider before deciding which approach to use. On the one hand, the mechanistic
modeling approach has the advantage of not requiring extensive fish tissue sampling and analysis by
using knowledge of aquatic system food webs and relatively simple measurements of selenium from the
aquatic system that are easier and less expensive to obtain. However, uncertainty can be introduced if
inappropriate parameters are chosen to model selenium bioaccumulation in the aquatic ecosystem. On the
other hand, the BAF approach is conceptually and computationally simpler because it is completely
empirical, relying only on field measurements with no need for any knowledge of the physical, chemical,
or biological characteristics of the waterbody. However, the BAF approach requires multiple
measurements of selenium in fish tissue that may be unavailable or expensive to obtain.
The appropriate modeling approach to use when translating the selenium egg-ovary FCV to a site-specific
water-column concentration depends on individual circumstances. Although the mechanistic modeling
approach may be a cost-effective method in situations where there is little or no current information about
selenium bioaccumulation, the BAF approach may be desirable in circumstances where substantial
resources have already been invested in fish tissue sampling and analysis. Because the national egg-ovary
selenium criterion is intended to apply to all waters of the United States, and site-specific BAF values or
the data required to derive site-specific BAF values are not available for the vast majority of those waters,
the EPA developed and utilized a translation methodology based on the mechanistic modeling approach.
Do not distribute, quote, or cite K-3 Draft Document
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Below is a description of how states and tribes may use this methodology to translate the egg-ovary FCV
to a site-specific water-column concentration for site-specific management of selenium.
1.1 Relating the Concentration of Selenium in Tissue and Water using the mechanistic modeling
approach
To relate the concentration of selenium in the eggs or ovaries of fish to the concentration of selenium in
the water-column, the EPA derived the equation:
C
f~i _ ^egg-ovary
water rrrrT^ composite T^T^ s~*T^
TTF p xEFxCF (Equation 18)
Cwater = the concentration of selenium in water (|ig/L),
C egg-ovary = the concentration of selenium in the eggs or ovaries of fish ((ig/g),
TTF°omposlte = the product of the trophic transfer function (TTF) values of the fish species that
is the target of the egg-ovary FCV and the TTF values of all lower trophic
levels in its food web (no units of measurement, see explanation below).
EF = the steady state proportional bioconcentration of dissolved selenium at the base
of the aquatic food web (L/g),
CF = the species-specific proportion of selenium in eggs or ovaries relative to the
average concentration of selenium in all body tissues (no units of
measurement),
The basic principles expressed in Equation 18 are illustrated in the conceptual model shown in Figure K-
1. Selenium dissolved in surface water enters aquatic food webs by becoming associated with trophic
level 1 primary producer organisms (e.g. algae) and other biotic (e.g. detritus) and abiotic (e.g. sediment)
particulate material. An enrichment function (EF) quantifies the bioconcentration of selenium in
particulate material and thus its bioavailability in the aquatic system. The parameter EF in Equation 18 is
a single value that represents the steady state proportional concentration of selenium in particulate
material relative to the concentration of selenium dissolved in water.
Particulate material is consumed by trophic level 2 organisms (usually aquatic invertebrates) resulting in
the accumulation of selenium in the tissues of those organisms. Trophic level 2 invertebrates are
consumed by trophic level 3 fishes resulting in further accumulation of selenium in the tissues offish.
Bioaccumulation of selenium from one trophic level to the next is quantified by a trophic transfer function
(TTF). A TTF is a single value that represents the steady state proportional concentration of selenium in
Do not distribute, quote, or cite K-4 Draft Document
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the tissue of an organism relative to the concentration of selenium in the food it consumes. Different
species of organisms metabolize selenium in different ways. Thus each species is associated with a
specific TTF value. Because the trophic transfer of selenium through all trophic levels is mathematically
equal to the product of the individual TTF values, all consumer-resource interactions in a particular
aquatic ecosystem are simplified in Equation 18 by representing the product of all the individual TTF
values as the single parameter TfF:omPos'te_
Fish accumulate selenium in different tissues of the body in differing amounts. Because the selenium
criterion is expressed as a concentration in the eggs and/or ovaries, a conversion factor (CF) quantifies the
relationship between the concentration of selenium in the eggs and/or ovaries and the average
concentration of selenium in the whole body. The parameter CFin Equation 18 is a single value that
represents the steady state proportional concentration of selenium in the eggs and/or ovaries relative to the
average concentration of selenium in all body tissues. Different species offish accumulate selenium in
their eggs and ovaries to different degrees. Thus each species offish is associated with a specific CF
value.
Do not distribute, quote, or cite K-5 Draft Document
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Egg-Ovary FC V
Species Egg-Ovary to Whole-Body Conversion Factor (CF)
Fish Whole-Body
Concentration
Species Trophic Transfer Function (TTF)
Invertebrate
Concentration
Species Trophic Transfer Function (TTF)
(J"ppcomposite\
Concentration in
Particulate Material
Enrichment Factor (EF)
Water-Column
Concentration
Figure K-l. Conceptual model for translating the egg-ovary FCV to a water-column concentration.
Once the parameters that quantify the transfer of selenium through each step in this pathway are
identified, they can be used with Equation 18 to translate the concentration of selenium in eggs and
ovaries to a concentration of selenium in the water-column.
Because each TTF value is species-specific, it is possible to differentiate bioaccumulation in different
aquatic systems by modeling the food web of the target fish species. For example, where the food web
contains more than 3 trophic levels, TTFcomposlte can be represented as the product of all TTF values for
each trophic level given as the following generalization of Equation 10 from the main text:
COmPOSite =
rr,rr,r,T
TTF'
-------
The consumption of more than one species of organism at the same trophic level can also be modeled by
expressing the TTF value at a particular trophic level as the average TTF values of all species at that
trophic level weighted by the proportion of species consumed given as:
TTFTLx = £ (TTF^ x w, ) (Equation 1 1)
i
where:
x _ ^e ^p^ transfer function of the ith species at a particular trophic level
= the proportion of the ith species consumed.
These concepts can be used to formulate a mathematical expression of ffpcomp°site faai models selenium
bioaccumulation in a variety of aquatic ecosystems. Figure K-2 illustrates four hypothetical food web
scenarios and the formulation of ffpcomp°site for each of them. For each scenario, the value of ffpcomp°site^
the CF value associated with the targeted fish species, and the site-specific EF value can be used with
Equation 9 to translate the egg -ovary FCV to a site-specific water concentration value. The general steps
required to derive a site-specific translation of the egg-ovary FCV are described below.
Do not distribute, quote, or cite K-7 Draft Document
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A) Three trophic levels (simple):
/7~"7~TI7' composite _ rr"T'T~'TL3 rr"T'T~'T
B) Four trophic levels (simple):
rj~"T'~C'C ompos i
'T"T'T~'TZ3
Three trophic levels (mix within trophic levels):
TTFc°mp°site = TTFT" x [(TTF?12 x W[)+ (iTF™ x w2)]
W2
D) Three trophic levels (mix across trophic levels):
rrrriT-'composite I rrirri T^ TL3
TTFTL3
w j
W
E) Four trophic levels (mix across trophic levels):
TTFc°mp°s"e = [(TTFT" x TTFT" x Wl)+(TTFT" x w2)]
777770
77777L2
/' 4
Figure K-2. Example mathematical expressions of rfrfpcomp°s'te representing different food-web scenarios.
jj,-pcompomte quan^a^veiy represents the trophic transfer of selenium through all dietary pathways of a
targeted fish species. The mathematical expression of the food-web model is used to calculate a value for
jj,-pcompomte usmg appropriate species-specific TTF values and the proportions of each species consumed at
each trophic level. See text for further explanation.
Do not distribute, quote, or cite
K-8
Draft Document
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1.2 Steps for deriving a site-specific water concentration value from the egg-ovary FCV
Below are the steps that can be taken to derive a site-specific water concentration value the selenium egg-
ovary FCV by completing the following steps:
1) Identify the appropriate target fish species.
2) Model the food-web of the targeted fish species.
3) Identify appropriate TTF values by either:
a. selecting the appropriate TTF values from a list of EPA-derived values, or
b. deriving TTF values from existing data, or
c. deriving TTF values by conducting additional studies, or
d. extrapolating TTF values from existing values.
4) Determine the appropriate value of EFby either
a. deriving a site-specific EF value from field measurements, or
b. deriving an appropriate EF value from existing data, or
c. extrapolating from EF values of similar waters, or
5) Determine the appropriate CF value by either,
a. selecting the appropriate CF value a list of EPA-derived values, or
b. deriving a CF value from existing data, or
c. deriving a CF value by conducting additional studies, or
d. extrapolating a CF value from existing values.
6) Translate the selenium egg-ovary FCV into a site-specific water concentration value using
Equation 9.
Below are detailed descriptions of each step followed by example calculations using a variety of
hypothetical scenarios.
1.2.1 Identify the appropriate target fish species
1.2.1.1 When fish are present
The EPA's selenium criterion is expressed as a concentration in the eggs and ovaries offish because the
selenium concentration in the eggs and ovaries is the toxicological endpoint of selenium and thus the
most sensitive and consistent indicator of toxicity. Nonetheless, the relationship between selenium
concentration in these tissues and selenium toxicity vary across species due to differences in sensitivity
and bioaccumulation potential. Therefore, states and tribes should choose the fish species that resides in
the aquatic system with the greatest risk of selenium toxicity.
Do not distribute, quote, or cite K-9 Draft Document
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The species most sensitive to selenium are those in the Salmonidae family. Thus, states and tribes should
target nonanadromous species in the Salmonidae family such as trout when they are present. Members of
the genus Lepomis (in the family Centrarchidae) that include bluegill are also sensitive and should be
targeted when no fish in the Salmonidae family are present. Other members of the Centrarchidae family
(such as bass) should be targeted if no fish of the genus Lepomis are present.
States and tribes should target nonanadromous species (species that do not migrate from salt water to
spawn in fresh water), because selenium exposure and subsequent bioaccumulation occurs over a
relatively long period of time through the consumption of locally contaminated aquatic organisms. If
nonanadromous fish species in the Salmonidae family is absent, states and tribes should target the
resident fish species likely to have the highest exposure and sensitivity to selenium. In aquatic systems
with resident fish species of unknown selenium sensitivity and bioaccumulation potential, factors such as
ecological significance can be factors in choosing which species to target. If the state or tribal monitoring
program uses lethal tissue sampling procedures, threatened or endangered species should not be used for
tissue monitoring.
Targeting fish species that consume organisms known or suspected to bioaccumulate selenium can be an
alternative approach to selecting fish species when species-specific information on selenium sensitivity
and bioaccumulation potential is unavailable. Prey organisms that pose a selenium toxicity risk to
predators usually have physiological characteristics that predispose them to selenium bioaccumulation
and/or are in close proximity to relatively high levels of selenium. For example, high levels of selenium
found in San Francisco Bay white sturgeon were linked to consumption of Potamocorbula amurensis, a
bivalve that was known to rapidly accumulate selenium and was in close proximity to selenium-
contaminated sediments (Stewart et al. 2004). In contrast, striped bass from the same aquatic system had
substantially lower levels of selenium due to their zooplankton-based food web with substantially lower
selenium bioaccumulation characteristics (Schlekat et al. 2004; Stewart et al. 2004).
States and tribes can use data from fisheries or biological surveys or other biological assessments to
determine the fish species that reside in specific surface waters. If such information is not available,
general information (often online) on the fish species that are present in state or tribal surface waters can
be found in:
• State Fish and Game agencies.
• U.S. Fish and Wildlife Service (http://www.fws.gov).
Do not distribute, quote, or cite K-10 Draft Document
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• U.S. Geological Survey (http://www.usgs.gov).
• NatureServe.org (http://www.natureserve.org).
• Fishbase (http://www.fishbase.org).
• State or local sources of biological information (e.g. Biota Information System of New Mexico at
http://www.bison-m.org).
Figure K-3 shows a decision tree that states and tribes may use to help select the appropriate fish species
for deriving a site-specific water concentration value from the selenium egg-ovary FCV. EPA
recommends this sequence of choices on the basis of taxonomic hierarchies that begin with taxa having
the highest sensitivity to selenium. The use of taxonomic hierarchies utilizes evolutionary relationships to
infer biological similarities among organisms (Suter 1993).
Do not distribute, quote, or cite K-ll Draft Document
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Yes
^
Are nonanadromous species of the
Salmonidae family present?
'
Target nonanadromous species in the
Salmonidae family (e.g. trout)
Yes
^
'
Target species in genus Lepomis (e.g.,
bluegill)
Yes
^
'
Target species in family Centrarchidae
(e.g. bass)
Yes
^
'
Target resident species with confirmed or
suspected sensitivity or exposure risk to
selenium.
No
1 r
Are species in the genus Lepomis
present?
No
1 r
Is family Centrarchidae present?
No
1 r
Are resident species with confirmed or
suspected sensitivity or exposure risk to
selenium present?
No/do not know
1 r
Target species with highest ecological
significance.
Figure K-3. Decision process for selection of the fish species to use when deriving a water concentration
from the selenium egg-ovary FCV.
1.2.1.2 When fish are absent
Some aquatic systems do not contain resident fish. Fish may be absent from a waterbody because of
intermittent or persistent low flows, physical impediments such as waterfalls or impoundments, lack of
adequate habitat for feeding and/or spawning, or intolerable aquatic conditions related to pH, turbidity,
temperature, salinity, total dissolved solids, chemical contaminants, or pathogens. These conditions could
be due to naturally occurring or anthropogenic causes. Some streams may be naturally intermittent or
ephemeral, or they might exhibit low or intermittent flows because of impoundments or water draw-down
for agricultural irrigation, industrial uses, drinking water supply, or other uses.
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K-12
Draft Document
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When fish are naturally absent from a waterbody, states and tribes should target the most sensitive fish
species inhabiting downstream waters. Although the upper reaches of some aquatic systems may not
support fish communities, the invertebrate organisms that reside there may tolerate high concentrations of
selenium and pose a selenium risk if they are transported downstream. In such cases, states and tribes may
still use the decision tree in Figure K-3 to identify downstream fish species to protect. In addition, states
and tribes may evaluate upstream waters without fish by measuring the selenium concentration in water,
biotic and/or abiotic particulate material, and/or the tissues of invertebrate aquatic organisms that reside
there. Because selenium associated with particulate material and invertebrate organisms can be
transported downstream during intermittent high flows, elevated concentrations of selenium in the tissues
of downstream fish could indicate upstream sources of selenium that require a more detailed evaluation of
upstream conditions. A site-specific selenium criterion protecting a limited aquatic environment may be
appropriate if selenium levels are naturally high and fish were not previously present in the aquatic
system.
1.2.2 Model the food-web of the targeted fish species
After selecting the target fish species, states and tribes should formulate a mathematical expression of the
target species food-web that will be used to calculate the value of TTFcomposlte. As discussed previously,
jj,-pcompomte -g ^ procjuct of me TTF values across trophic levels of the target fish species food-web. The
complexity of the food-web model will depend on the species offish that is targeted, the diversity of prey
species in the aquatic system, and the amount of information that is available. Many of the same
information sources used to identify the targeted fish species in a waterbody might also be used to obtain
information about its food web. The types and proportions of food organisms consumed by the targeted
fish species can be directly determined through studies that examine stomach contents, or from
information gathered through biological assessments. If site-specific, field-derived information is not
available, the food-web characteristics of the target species can be estimated using publicly available
databases such as NatureServe (http://www.natureserve.org). For example, in the HUC watershed
#5040004 in Ohio, the NatureServe database record for fathead minnow indicates under the heading:
"Ecology and Life History - Food Comments," the fathead minnow "feeds opportunistically in soft
bottom mud; eats algae and other plants, insects, small crustaceans, and other invertebrates (Becker 1983,
Sublette et al. 1990)."
Do not distribute, quote, or cite K-13 Draft Document
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Additional sources of information include:
• FishBase (http://www.fishbase.org). FishBase is a relational database developed at the World
Fish Center in collaboration with the Food and Agriculture Organization of the United Nations
(FAO) and many other partners.
• Carlander, K.D. Handbook of Freshwater Fishery Biology, volumes 1, 2 and 3. Iowa state
University Press, Ames, Iowa. 1969-1997.
1.2.3 Identify appropriate TTF values
The food-web model uses appropriately selected species-specific TTF values (and, if appropriate,
proportions within the same trophic level). States and tribes can determine the appropriate TTF values to
calculate ffpcomP°slte by either using one of the following four procedures, or by using other scientifically
defensible methods.
1.2.3.1 Select the appropriate TTF values from a list of EPA-derived values
Species-specific TTF values represent the steady state proportional concentration of selenium in the tissue
of an organism relative to the concentration of selenium in the food it consumes. TTF values for aquatic
invertebrates and fish are provided in Tables K-l and K-2 (see main text for a complete explanation of
how these values were derived).
Table K-l. EPA-derived Trophic Transfer Function (TTF) values for freshwater aquatic
invertebrates.
Common name
Crustaceans
amphipod
copepod
crayfish
water flea
Insects
dragonfly
damselfly
mayfly
midge
water boatman
Mollusks
asian clama
zebra mussel
Scientific name
Hyalella azteca
Copepods
Astacidae
Daphnia magna
Anisoptera
Coenagrionidae
Centroptilum triangulifer
Chironomidae
Corixidae
Corbicula fluminea
Dreissena polymorpha
AE IR ke TTF
1.22
0.520 0.420 0.155 1.41
1.46
0.406 0.210 0.116 0.74
1.97
2.88
2.38
1.90
1.48
0.550 0.050 0.006 4.58
0.260 0.400 0.026 4.00
Do not distribute, quote, or cite K-l4 Draft Document
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Common name Scientific name AE IR ke TTF
Annelids
blackworm Lumbriculus variegatus 0.165 0.067 0.009
Other
zooplankton Zooplankton -
3 Not to be confused with Corbula amurensis
Table K-2. EPA-derived Trophic Transfer Function (TTF) values for freshwater fish
Common name Scientific name AE IR ke
Cypriniformes
bluehead sucker Catostomus discobolus
common carp Cyprinus carpio
creek chub Semotilus atromaculatus
fathead minnow Pimephales promelas
flannelmouth sucker Catostomus latipinnis
longnose sucker Catostomus catostomus
sand shiner Notropis stramineus
white sucker Catostomus commersonii
Cyprinodontiforme s
mosquitofish Gambusia sp. ...
northern plains killifish Fundulus kansae
western mosquitofish Gambusia affinis
Esociformes
northern pike Esox lucius
Gasterosteiformes
brook stickleback Culaea inconstans
Perciformes
black crappie Pomoxis nigromaculatus
bluegill Lepomis macrochirus
green sunfish Lepomis cyanellus
largemouth bass Micropterus salmoides
striped bass Morone saxatilis 0.375 0.335 0.085
walleye Sander vitreus
yellow perch Percaflavescens
Salmoniformes
brook trout Salvelinus fontinalis
1.29
1.89
TTF
1.04
1.34
1.12
1.57
1.06
0.90
1.83
1.18
0.86
1.27
1.25
2.04
1.69
2.67
1.48
1.27
1.27
1.48
1.82
1.42
0.88
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K-15
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Common name Scientific name AE IR ke TTF
brown trout Salmo trutta - - - 1 .44
cutthroat trout Oncorhynchus clarkii • • - 1.07
mountain whitefish Prosopium williamsoni • • • 1.38
rainbow trout Oncorhynchus mykiss • • • 1.19
westslope cutthroat trout Oncorhynchus clarkii lewisi - - - 1.20
Scorpaeniformes
mottled sculpin Coitus bairdi • • • 1.38
sculpin Coitus sp. ... 1 .29
Siluriformes
black bullhead Ameiurus melas • • • 0.91
channel catfish Ictalurus punctatus • • • 0.73
The TTF values from these lists could be used exclusively, or in conjunction with TTF values obtained
from other sources (see below). Note that these tables do not represent an exhaustive list of all TTF values
that may be required to calculate a site-specific water concentration value. If this list does not include a
required TTF value, states and tribes should refer to other approaches to obtain an appropriate value.
1.2.3.2 Deriving TTF values from existing data
If one or more appropriate TTF values cannot be found in Tables K-l and/or K-2, states and tribes could
derive species-specific TTF values using existing data. One approach for deriving species-specific TTF
values is to use the physiological coefficients representing food ingestion rate (IR), selenium efflux rate
(ke), and selenium assimilation efficiency (AE) to calculate a TTF value using Equation 3.
If the TTF value of a particular species in a food web is not available, TTF may be derived in several
different ways. One method is to obtain the physiological coefficients of food ingestion rate (IR),
assimilation efficiency (AE), or efflux rate (ke) and apply those values to Equation 3 given as:
(Equation 3)
Where:
TTF = species-specific trophic transfer function
AE = species-specific assimilation efficiency (%)
IR = species-specific ingestion rate (g/g-d)
ke = species-specific efflux rate constant (/d)
Do not distribute, quote, or cite K-l6 Draft Document
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The physiological coefficients IR, AE and ke may be obtained from published literature or may be derived
from laboratory studies. Another way to derive species-specific TTF values is to empirically assess the
relationship between the selenium concentration in the tissue of organisms and the selenium concentration
in the food they consume using measurements from field studies. Species-specific TTF values can be
derived from such measurements by calculating ratios, using regression techniques, or other scientifically
defensible methods.
The physiological coefficients AE, IR, and ke are species-specific values. Coefficients AE and ke can only
be derived from laboratory experiments, butIR can be derived from either laboratory or field studies.
After the three physiological coefficients are obtained, a TTF value can be calculated using Equation 3
from the main text. Another approach for deriving species-specific TTF values is to use paired selenium
measurements of consumer organisms and their potential resources from field studies that directly
measure the trophic transfer of selenium in those organisms. The TTF for any trophic level can be defined
by the equation:
rTLn
TTFTLn = tz&se. (Equation K-l)
Cfood
Where:
TTpTLn _ jhe trophic transfer function of a given trophic level,
Ctissue = The selenium concentration (mg/kg dw) in the tissues of the consumer
organism,
selenium concentration (mg/kg dw) in the consumer organism's food.
At a given site, the empirical relationship between an organism and its food is first confirmed with linear
regression analysis. If the regression is both statistically significant (P < 0.05) and the slope of the
relationship is positive (i.e., selenium concentrations in the consumer increases with increasing selenium
in food), then the data are considered acceptable and the species-specific TTF is determined as the median
ratio of the paired consumer-food selenium concentration data. Both of the above methods were used to
derive the TTF values provided in Tables K-l and K-2.
1.2.3.3 Deriving TTF values by conducting additional studies
States and Tribes may conduct additional studies to collect the data needed to derive TTF values for
specific needs or to revise existing TTF values. TTF values could be derived from new data using the
methodology described above, or other scientifically defensible methods. If available TTF values do not
apply to the species in a waterbody, there are no site-specific data available, and the collection of
Do not distribute, quote, or cite K-l7 Draft Document
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necessary data by a state is impractical; then the state could require data collection where appropriate
based on Section 308 of the CWA (or comparable State authority).
1.2.3.4 Extrapolating TTF values from existing values
When one or more TTF values are not available for a species, and the information needed to derive a
species-specific TTF value is not available or impractical to obtain, a TTF value can be extrapolated from
known TTF values. One possible method to extrapolate a TTF value is to sequentially consider higher
taxonomic classifications until one or more of the organisms with a known TTF value matches the taxon
being considered. If the lowest matching taxon is common to more than one of the available TTF values,
the average TTF from the matching table entries could be used. The use of taxonomic hierarchies in this
way utilizes evolutionary relationships to infer biological similarities among organisms (Suter 1993).
EPA used this extrapolation procedure to derive the TTF values of some of the organisms at some
representative aquatic sites that were used to derive the recommended water concentration values for lotic
and lentic waters. For example, the TTF value for Gila robusta, the roundtail chub, was not listed in Table
K-2. However, the roundtail chub is in the family Cyprinidae, which includes TTFs for common carp,
creek chub, fathead minnow, and sand shiner. Because Cyprinidae is the lowest taxonomic classification
where the fish species being considered matches a taxon in Table K-2, the median TTF value for the
family Cyprinidiae (1.46) was used for the roundtail chub.
1.2.4 Determine the appropriate EF value
The selenium enrichment function EF represents the bioavailability of selenium at the base of the aquatic
food web. The parameter EF varies more widely across aquatic systems than any other parameter, and is
influenced by the source and form of selenium, water residence time, and the biogeochemical
characteristics of the waterbody. Because EF can vary greatly across waterbodies, this parameter has the
greatest potential to introduce uncertainty in the translation from an egg-ovary concentration of selenium
to a water concentration and should be considered carefully. States and tribes can determine an
appropriate EF value either by using one of the following four procedures, or by using other scientifically
defensible methods.
Do not distribute, quote, or cite K-18 Draft Document
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1.2.4.1 Deriving a site-specific EF value from field measurements
The parameter EF can be expressed as the ratio of the concentration of selenium in particulate material to
the concentration of selenium dissolved in water. Using the equation:
Cr
7^7^ -'particulate . . ,-,
EF = — (Equation 12)
water
Where:
C
particulate = Concentration of selenium in particulate material ((^g/g)
water = Concentration of selenium dissolved in water ((ig/L)
EF = Enrichment Function (L/g)
Deriving a site-specific EF value in this manner is a relatively straightforward and inexpensive procedure.
An EF value for a particular aquatic system can be derived by collecting water samples, separating the
particulate material from the water in each sample, measuring the concentration of selenium in the
separated water and particulate material, computing the ratio of the two measurements from each sample,
and then calculating the mean or median of all the ratios. Alternatively, a state or tribe could derive an EF
value by verifying the statistical relationship between paired particulate and water concentrations through
linear regression, calculating the ratios of paired Cparticuiate to Cwater values, and then taking the median
ratio as the value of EF. This approach statistically evaluates the data representing the relationship
between these two media. However, a sufficient quantity of data is necessary to calculate statistically
significant fits.
Regardless of the method used to derive the value of EF from field measurements, field and analytical
methods should be carefully planned and implemented when developing a site-specific, field-derived EF
value. Selenium bioaccumulation occurs more readily in aquatic systems with longer residence times
(such as wetlands, oxbows, and estuaries) and with fine particulate sediments high in organic carbon.
Thus EPA recommends a sampling plan that prioritizes areas with these characteristics. Analytical
methods to measure selenium in particulate material and in water are discussed in Appendix L.
1.2.4.2 Deriving an appropriate EF value from existing data
If suitable and sufficient site-specific measurements of Cparticuiate and Cwater are available, states and tribes
could derive an EF value using these data. However, states and tribes should ensure that these data
represent current conditions, are based on scientifically acceptable sampling techniques, and are obtained
using proper quality assurance and quality control protocols to minimize uncertainty.
Do not distribute, quote, or cite K-19 Draft Document
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1.2.4.3 Extrapolating from EF values of similar waters
In circumstances where a site-specific, field-derived EF value is not available, an EF value from one or
more aquatic systems with similar hydrological, geochemical, and biological characteristics could be used
to approximate EF. However, states and tribes should carefully consider the possibility of introducing
significant uncertainty into the calculation when using EF values extrapolated from other waterbodies.
States and tribes should not use EF values derived from large-scale sites that encompass multiple water
bodies or ecosystems, or that do not match the characteristics of the waterbody for which the water
column concentration of selenium is being derived.
1.2.5 Determine the appropriate CF value.
1.2.5.1 Selecting the appropriate CF value from the list of values that were used to derive EPA's
recommended water criteria concentration values.
The parameter CF represents the species-specific proportion of selenium in eggs or ovaries relative to the
average concentration of selenium in all body tissues. EPA derived species-specific CF values for 17
species of fish from studies that measured selenium concentrations in both eggs and/or ovaries and in
whole body and/or muscle. These CF values can be found in Appendix B and are reproduced below
(Table K-3).
Table K-3. Whole Body Se to Egg-Ovary Se Conversion Factors (CF)
Common name
Species
Bluegill
Bluehead sucker
Brook trout
Brown trout
Common carp
Cutthroat trout
Dolly varden
Flannelmouth sucker
Green sunfish
Mountain whitefish
Northern pike
Median ratio
y^egg-ovary ^whole-body)
2.13
1.82
1.45
1.92
1.96
1.41
1.45
Median ratio
\\^egg-ovary ^muscle)
1.09
1.26
5.80
1.88
Muscle to
whole-body
correction
factor
1.27
1.27
1.27
1.27
Final CF
values
2.13
1.82
1.38
1.45
1.92
1.96
1.61
1.41
1.45
7.39
2.39
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K-20
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Common name
Rainbow trout
Razorback sucker
Roundtail chub
Smallmouth bass
White sturgeon
White sucker
Median ratio
V ^egg-ovary ^whole-body)
2.07
1.42
1.41
Median ratio
V ^egg-ovary ^muscle)
1.92
1.12
1.33
Muscle to
whole-body
correction
factor
1.27
1.34
1.27
Final CF
values
2.44
1.51
2.07
1.42
1.69
1.41
Genus
Catostomus
Esox
Lepomis
Micropterus
Oncorhynchus
1.41
2.39
1.79
1.42
1.96
Family
Catostomidae
Centrarchidae
Cyprinidae
Salmonidae
1.41
1.45
2.00
1.96
Order
Perciformes
1.45
Class
Actinopterygii
1.63
The data and methods used to derive CF for these species are described in Appendix B.
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K-21
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1.2.5.2 Deriving a CF value from existing data
The parameter CFis mathematically expressed as:
c
CF =
egg-ovary
wMe-body (Equation 16)
where
CF = Whole-body to egg-ovary conversion factor (dimensionless ratio).
Cegg-ovary = Selenium concentration in the eggs or ovaries offish (|ig/g)
Cwhoie-body = Selenium concentration in the whole body offish (mg/kg).
If suitable and sufficient data are available, a state or tribe could derive a species-specific CF value using
the same numerical methods used to calculate the parameter EF. A state or tribe could calculate the ratio
of the two concentrations in each sample tissue as defined in Equation 16, and then calculate the mean or
median of all the ratios. Alternatively, a state or tribe could derive a CF value by verifying the statistical
relationship between paired particulate and water concentrations through linear regression, calculating the
ratios of paired Cem_mary to Cwhole_body, and then taking the median ratio of the paired values as the CF (see
Appendix B). This approach statistically evaluates the data representing the relationship between these
two tissue types. However, a sufficient quantity of data is necessary to calculate statistically significant
fits. Regardless of the method used, care should be taken to ensure that the data used accurately represents
current conditions, were based on scientifically acceptable sampling techniques, and were obtained using
acceptable quality assurance and quality control protocols.
1.2.5.3 Deriving a CF value by conducting additional studies
States and tribes could perform additional studies to obtain the data needed to derive CF values for
specific needs or to revise existing CF values if there is reason to believe doing so may increase the
accuracy of the resulting water concentration. Analytical methods to measure selenium in tissue are
discussed in Appendix L. Where appropriate, additional data could be obtained as part of a NPDES
permit application by invoking authority under CWA section 308 (or comparable State authority) to
reasonably require NPDES-regulated facilities to collect information necessary to develop NPDES permit
limits.
1.2.5.4 Extrapolating the CF value from the list of values that were used to derive EPA's
recommended water criteria concentration values
Because the pattern of selenium concentration in different body tissues varies between species,
extrapolating a species-specific CF value from one or more surrogate species is not recommended.
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However, a CF value that is the average of all known species-specific CF values could be used when the
CF value of the target species is not available, and the data needed to derive a species-specific CF value is
not available or impractical to obtain. Using the average species CF value, however, lowers translation
accuracy and should only be used when other species-specific options are not available.
1.2.6 Translate the selenium egg-ovary FCV into a site-specific water concentration value using
Equation 18.
After determining the appropriate values of CF, TTFcomposlte (derived from the product of the individual
TTF values from each trophic level) and EF, a site-specific water concentration can be derived from the
egg-ovary FCV using Equation 18 of the main text. Note that NPDES permitting regulations at 40 CFR §
122.45(c) requires that a Water Quality-Based Effluent Limit (WQBEL) for metals be expressed as total
recoverable metal, unless an exception is met under 40 CFR § 122.45(c)(l)-(3). Equation 18 assumes
selenium concentrations dissolved in water. While states and tribes may express ambient water quality
criteria in water quality standards as dissolved selenium, an additional step is necessary to convert the
dissolved selenium concentration to a total recoverable selenium concentration for the purpose of NPDES
permitting. Guidance for converting expression of metal concentrations in water from dissolved to total
recoverable can be found in Technical Support Document for Water Quality-based Toxics Control (U.S.
EPA 1991) and The Metals Translator: Guidance for Calculating a Total Recoverable Permit Limit from
a Dissolved Criterion (U.S. EPA 1996).
1.3 Managing uncertainty using the mechanistic modeling approach
Derivation of a water concentration from the egg-ovary FCV using the mechanistic bioaccumulation
modeling approach (Equation 18) is subject to uncertainties from several sources. These include:
• Measurement error when deriving input parameters.
• Unaccounted factors affecting bioaccumulation.
• Inaccurate identification or proportions of trophic level 2 food-web organisms.
• Inaccurate or inappropriate TTF, EF, or CF values.
• Biological variability.
• Other unknown factors.
Though not required, the effectiveness of effluent limits and waste load alsites of selenium that are based
on water concentration values derived from the egg-ovary FCV should be confirmed whenever practical
using appropriate fish tissue assessment methods. In addition, comparing estimated selenium
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concentrations in fish tissue with actual selenium concentrations obtained from small-scale field studies
could also help evaluate the suitability of selected equation parameters.
1.4 Example calculations
Below are six hypothetical examples that demonstrate how to translate the egg-ovary FCV to a site
specific water concentration criterion using Equation 18. These examples encompass a variety of
hypothetical aquatic systems with various fish species and food webs. For these hypothetical examples,
species-specific TTF values were taken from Tables K-l and K-2, and CF values were taken from Table
K-3. To calculate EF in these examples, the EPA used a hypothetical water concentration of 5 (ig/L and
the hypothetical particulate concentrations of 4.25 (ig/g and 8.75 (ig/g in lotic and lentic aquatic systems,
respectively.
1.4.1 Example 1
Bluegill (Lepomis macrochirus) in a river that consume mostly amphipods:
Current water concentration ((ig/L)
Current particulate concentration (mg/kg)
Trophic transfer function for bluegill (TTF113)
Trophic transfer function for amphipods (TTF™)
Egg -ovary to whole-body conversion factor for bluegill (CF)
Selenium egg-ovary FCV (mg/kg)
5.00
4.25
1.48
1.22
2.13
15.8
EF =
EF =
4.25
^particulate
5.00
= 0.85 L/g
^egg-ovary
X EF X CF
Combined rprpj^TLB .. rprpj^TL2
= 1.48 x 1.22
= 1.81
15.8
^water ~
1.81 xO.85 X2.13
= 4.82(ig/L
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1.4.2 Example 2
Fathead minnow (Pimephales promelas) in a river that consume mostly copepods:
Current water concentration ((ig/L)
Current particulate concentration (mg/kg)
Trophic transfer function for fathead minnow (TTF113)
Trophic transfer function for copepods (TTFTL2)
Egg -ovary to whole-body conversion factor for fathead minnow (species-specific value
not available, so median CF for family Cyprinidae is used) (CF)
Selenium egg-ovary FCV (mg/kg)
5.00
4.25
1.57
1.41
2.00
15.8
„„ ^particulate .„ . „,
EF = — (Equation 12)
EF =
4.25
5.00
= 0.85 L/g
^egg-ovary
_ - ,„ „ _,
vater = TTF combined x Ep x Cp (Equation 18)
rrirri-r-COmbined _ 'T-"T-'T^TL3 .. IT"T'T
= 1.57 x 1.41
= 2.21
15.8
water =
= 4.21
2.21x0.85x2.00
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1.4.3 Example 3
Bluegill (Lepomis macrochirus) in a lake that consume mostly aquatic insects:
Current water concentration ((ig/L)
Current particulate concentration (mg/kg)
Trophic transfer function for Bluegill (TTFTL3)
Trophic transfer function for aquatic insects (median of Odonates, Water boatman,
Midges, and Mayflies) (TTFTL2)
Egg-ovary to whole-body conversion factor for Bluegill (CF)
Selenium egg-ovary FCV (mg/kg)
5.0
8.75
1.48
2.17
2.13
15.8
EF =
particulate
EF =
8.75
5.00
= 1.75L/g
,„ . . „.
(Equation 12)
x CF
^water ~
= 1.48x2.17
= 3.21
15.8
3.21x1.75x2.13
= 1.32ng/L
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1.4.4 Example 4
Fathead minnow (Pimephales promelas) in a river that consume approximately % copepods and
aquatic insects:
Current water concentration ((ig/L)
Current particulate concentration (mg/kg)
Trophic transfer function for fathead minnow (TTF113)
Trophic transfer function for copepods and aquatic insects (TTFTL2)
Copepods =1.41
Average of all aquatic insects = 2.17
^(TTF, xw,)
XTFTL2 = i=l
= (1.41 x%) + (2.17x i/3)
= 1.66
Egg -ovary to whole-body conversion factor for fathead minnow (species-specific value
not available, so median CF for family Cyprinidae is used). (CF)
Selenium egg-ovary FCV (mg/kg)
5.0
4.25
1.57
1.66
2.00
15.8
EF =
EF =
^particulate
^water
4.25
(Equation 12)
5.00
= 0.85L/g
^egg-ovary
TTFcombined x Ep x Cp
(Equation 18)
ined = J"pfTL3 x TTF112
= 1.57x1.66
= 2.61
15.8
^water ~
2.61x0.85x2.00
= 3.56fig/L
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1.5.5 Example 5
Fathead chub (Platygobio gracilis) in a river with a diet of approximately 80% aquatic insects and
20% algae:
Current water concentration ((ig/L)
Current particulate concentration (mg/kg)
Trophic transfer function of fathead chub:
Lowest matching taxon is the family Cyprinidae. Therefore, the TTF value of
Cyprinidae is used (TTF113)
Trophic transfer function for insects (TTFTL2)
Average of all aquatic insects = 2.17
Egg -ovary to whole-body conversion factor for fathead chub (species-specific value not
available, so median CF for family Cyprinidae is used). (CF)
Selenium egg-ovary FCV (mg/kg)
5.0
4.25
1.46
2.17
2.00
15.8
„„ ^particulate ,„ . _N
EF = -J—- (Equation 12)
EF =
^water
4.25
5.00
= 0.85L/g
TTF combined = [TTFTL3 x TTpTL2
Where:
[TTF
TL3
wi = Proportion of fathead chub diet from insects; and
W2 = Proportion of fathead chub diet from algae
TTFcomb = [Ii46 x 2.17 x 0.8] + [1.46 X 0.2]
= 2.83
15.8
water ~ 2.83 X0.85 X2.00
= 3.28ng/L
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1.5.6 Example 6
Largemouth bass (Micropterus salmoides) in a large river that consume mostly Western
mosquito fish (Gambusia affinis) that consume approximately % insects and % crustaceans:
Current water concentration ((ig/L)
Current particulate concentration (mg/kg)
Trophic transfer function of largemouth bass (TTFTL4)
Trophic transfer function of Western mosquitofish (TTFTL3)
Trophic transfer function for insects and crustaceans (TTF112)
Median all Insects -2.11
Median all Crustaceans - 1.41
±(TTF^WI)
XTFTL2 = '=1
= (2.17 x3/4) + (1.41 x%)
= 1.98
Egg-ovary to whole-body conversion factor for Largemouth bass (species-specific
value not available, so median for the genus Micropterus used) (CF)
Selenium egg-ovary FCV (mg/kg)
5.0
4.25
1.27
1.25
1.98
1.42
15.8
EF =
^particulate
(Equation 12)
EF =
4.25
5.00
= 0.85L/g
ffpcombined = J"pfTL4 x TTF?13 X TTF1"12
= 1.27x1.25x1.98
= 3.14
15.8
'water
3.14x0.85x1.42
= 4.17(ig/L
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1.5.7 Example 7a
Derivation of a site specific water column criterion for a river impacted by selenium
Available data for a site indicates that the average egg/ovary tissue concentration of selenium for the
bluegill (Lepomis macrochirus) is 22 mg/kg (dw). This concentration exceeds the USEPA proposed
egg/ovary criterion of 15.8 mg/kg (dw). The translated selenium water column criterion for lotic
waterbodies is 4.2 ug/L. The following calculation shows how to derive a water column concentration
that would achieve the 15.8 mg/kg (dw) egg/ovary tissue criterion.
Site specific selenium egg/ovary concentration (bluegill; mg/kg dw)
Selenium egg/ovary criterion (mg/kg, dw)
Selenium Water Column Criterion, Lotic Habitats (ug/L)
Allowable lotic water column concentration (ug/L)
22.0
15.8
4.2
X
1. Set up proportional equation to solve for allowable water column concentration
Lotic Water Column Criterion
Allowable Water concentration (X)
Current egg/ovary FT concentration
Selenium egg/ovary criterion
4.2 ug/L
X
22 mg/kg dw
15.8 mg/kg dw
X
X
4.2x 15.8
22
66.36
22
3.02 ug/L = Target Site specific Lotic Water Column Criterion
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1.5.7 Example 7b
Derivation of a site specific water column criterion for a lake impacted by selenium
Available data for a site indicates that the average egg/ovary tissue concentration of selenium for the
bluegill (Lepomis macrochirus) is 22 mg/kg (dw). This concentration exceeds the USEPA proposed
egg/ovary criterion of 15.8 mg/kg (dw). The translated selenium water column criterion for lentic
waterbodies is 0.9 ug/L. The following calculation shows how to derive a water column concentration
that would achieve the 15.8 mg/kg (dw) egg/ovary tissue criterion.
Current selenium egg/ovary concentration (bluegill; mg/kg dw)
Selenium egg/ovary criterion (mg/kg, dw)
Selenium Water Column Criterion, Lentic Habitats (ug/L)
Allowable lotic water column concentration (ug/L)
22.0
15.8
0.9
X
2. Set up proportional equation to solve for allowable water colun concentration
Lentic Water Column Criterion
Allowable Water concentration (X)
Current egg/ovary FT concentration
Selenium egg/ovary criterion
0.9 ug/L
X
22 mg/kg dw
15.8 mg/kg dw
X
X
0.9x 15.8 = 14.22
22 22
0.65 ug/L = Target Site specific Lentic Water Column Criterion
2.0 TRANSLATING THE CONCENTRATION OF SELENIUM IN TISSUE TO
A CONCENTRATION IN WATER USING BlOACCUMULATION FACTORS
(BAF).
2.1 Summary of the BAF approach
A bioaccumulation factor (BAF) is the ratio (in milligrams/kilogram per milligrams/liter, or liters per
kilogram) of the concentration of a chemical in the tissue of an aquatic organism to the concentration of
the chemical dissolved in ambient water at the site of sampling (U.S. EPA 2001c). BAFs are used to
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relate chemical concentrations in aquatic organisms to concentrations in the ambient media of aquatic
ecosystems where both the organism and its food are exposed and the ratio does not change substantially
overtime. The BAF is expressed mathematically as:
BAF=C^ue_
r
water (Equation J-2)
where
BAF = bioaccumulation factor derived from site-specific field-collected samples of
tissue and water (L/kg)
Cttssue = concentration of chemical in fish tissue (mg/kg)
CWater = ambient concentration of chemical in water (mg/L)
Solving for Cwater:
/"< tissue /T^ r , i
L water ~ TTTTT (Equation J-3)
To translate a fish tissue criterion to a water concentration value, states and tribes could develop a site-
specific, field-measured BAF for the waterbody, and then calculate a water concentration criterion using
Equation J-3. Detailed information about how to derive a site-specific, field-measured BAF is provided in
Methodology for Deriving Ambient Water Quality Criteria for the Protection of Human Health (2000)
Technical Support Document Volume 3: Development of Site-specific Bioaccumulation Factors (U.S.
EPA 2009). Although this guidance was developed for deriving human health criteria, the methodological
approach is also applicable to the derivation of aquatic life criteria.
2.2 Managing uncertainty using the BAF approach
Considerable uncertainty can be introduced when using the BAF approach to derive a water concentration
value from a fish tissue criterion concentration. Inaccurate water concentration values can result when
BAFs are derived from water and fish tissue concentration measurements that are obtained from sources
that do not closely represent site characteristics, or from field data collected from large-scale sites that
encompass multiple water bodies or ecosystems. Most of this uncertainty results from differences in the
bioavailability of selenium between the study sites where measurements are made to derive the BAF, and
the site(s) to which the BAF is used to derive needed water concentration values.
Because of uncertainties associated with the BAF approach, EPA does not recommend developing BAFs
from data extrapolated from different sites or across large spatial scales. EPA's Framework for Metals
Risk Assessment (U.S. EPA 2007) outlines key principles about metals and describes how they should be
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considered in conducting human health and ecological risk assessments. The current science does not
support the use of a single, generic threshold BAF value as an indicator of metal bioaccumulation. The
use of BAFs are appropriate only for site-specific applications where sufficient measurements have been
taken from the site of interest and there is little or no extrapolation of BAF values across differing
exposure conditions and species.
The preferred approach for using a BAF to implement the selenium fish tissue criterion is to calculate a
site-specific, field-measured BAF from data gathered at the site of interest, and to apply that BAF to that
site. A site-specific, field-measured BAF is a direct measure of bioaccumulation in an aquatic system
because the data are collected from the aquatic ecosystem itself and thus reflects real-world exposure
through all relevant exposure routes. A site-specific, field-measured BAF also reflects biotic and abiotic
factors that influence the bioavailability, biomagnification, metabolism, and biogeochemical cycling of
selenium that might affect bioaccumulation in the aquatic organism or its food web. Appropriately
developed site-specific, field-measured BAFs are appropriate for all bioaccumulative chemicals,
regardless of the extent of chemical metabolism in biota from a site (U.S. EPA 2000).
Although a site-specific, field-measured BAF is a direct measure of bioaccumulation, its predictive power
depends on a number of important factors being properly addressed in the design of the field sampling
effort. For example, sampling in areas with relatively long water residence times should be a priority
because selenium bioaccumulation occurs more readily in aquatic systems with longer residence times
(such as wetlands, oxbows, and estuaries) and with fine particulate sediments high in organic carbon. In
addition, migratory species should generally not be used because their exposure to selenium could reflect
selenium concentrations in areas other than where the fish were caught. Fish may also need to be sampled
and BAF values recalculated if selenium levels significantly change overtime because BAFs are known
to be affected by the ambient concentration of the metals in the aquatic environment (McGeer et al. 2003;
Borgman et al. 2004; DeForest et al. 2007). States and tribes should refer to Methodology for Deriving
Ambient Water Quality Criteria for the Protection of Human Health (2000) Technical Support Document
Volume (U.S. EPA 2009) for guidance on appropriate methods for developing a site-specific, field-derive
BAF.
The advantage of using the BAF approach is its relative simplicity, especially when the data necessary to
derive the BAF is already available. Furthermore, the BAF approach is completely empirical and does not
require any specific knowledge about the physical, chemical, or biological characteristics of the
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waterbody. The relationship between the concentration of selenium in fish tissue and water is directly
determined by direct measurements in these media.
Limitations of the BAF approach should be considered before deciding if this method is appropriate for
translating the selenium FCV to a water concentration value. One disadvantage of the BAF approach is
the considerable time and cost necessary to collect sufficient data to establish the relationship between
tissue and water concentrations. Costs increase as the spatial scale and complexity of the aquatic system
increases. Furthermore, the BAF approach does not allow extrapolation across species, space, and large
time scales because the site-specific factors that might influence bioaccumulation are integrated within
the tissue concentration measurements and thus cannot be individually adjusted to extrapolate to other
conditions. Thus, site-specific, field-measured BAFs only provide an accounting of the uptake and
accumulation of selenium for an organism at a specific site and point in time.
As noted previously, NPDES permitting regulations at 40 CFR § 122.45(c) require WQBELs for metals
be expressed as total recoverable metal unless an exception is met under 40 CFR § 122.45(c)(l)-(3).
Guidance for converting expression of metals in water from dissolved to total recoverable can be found in
Technical Support Document for Water Quality-based Toxics Control (U.S. EPA 1991) and TheMetals
Translator: Guidance for Calculating a Total Recoverable Permit Limit from a Dissolved Criterion (U.S.
EPA 1996). Whether or not a water concentration value derived from a site-specific, field-derived BAF
requires conversion from dissolved to total recoverable selenium depends on how the BAF is developed.
Generally, conversion would not be necessary if the BAF is derived from water concentration values that
measure total selenium; however, conversion would be necessary if the BAF was derived from water
concentration values that measured dissolved selenium. Table K-4 compares some of the principle
characteristics of the mechanistic bioaccumulation modeling approach or the BAF approach for
translating the selenium FCV to a water concentration.
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3.0 COMPARISON OF MECHANISTIC BIOACCUMULATION MODELING
AND BAF APPROACHES
Table K-4. Comparison of mechanistic bioaccumulation and BAF approaches.
Mechanistic bioaccumulation modeling
Bioaccumulation Factor (BAF)
Knowledge of the aquatic system needed
Choice of input parameters at discretion of State or
Tribe
Species-specific
Can be applied at different sites
Fish tissue sampling not required for translation
No information on aquatic system needed
No input parameters to choose
Species-specific
Site-specific
Fish tissue sampling required
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APPENDIX L: ANALYTICAL METHODS FOR
MEASURING SELENIUM
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The Clean Water Act (CWA) establishes an EPA approval process for certain analytical methods used in
the National Pollutant Discharge Elimination System (NPDES) program and for section 401
certifications. EPA has several approved methods for measuring selenium in water under 40 CFR § 136.
EPA generally requires the use of EPA-approved methods for the NPDES program and for CWA section
401 certifications issued by states and tribes (40 CFR § 136.1). However, since there are no EPA
approved methods for the analysis of selenium in fish tissue, states and tribes may use analytical methods
not approved by EPA to evaluate the attainment of water quality standards or to develop or implement
Total Maximum Daily Loads provided that these methods are scientifically sound (40 CFR 122.21(g)(7)).
Implementation of a water quality standard for selenium may require the ability to detect and measure the
concentration of selenium in effluent, ambient water, tissue, and other media that is below the detection
limit or limit of quantitation that some analytical methods can provide. States and tribes should choose an
analytical method that is sufficiently sensitive to implement its water quality standard for selenium.
Below are descriptions of some of the methods available for measuring selenium concentrations with
sufficient sensitivity to implement EPA's recommended selenium criterion. Complete descriptions of
analytical methods appropriate for analyzing selenium in different media can be found in the National
Environmental Methods Index at http://www.nemi.gov.
GENERAL CONSIDERATIONS WHEN MEASURING CONCENTRATIONS OF
SELENIUM
The oxidation states of selenium dissolved in surface water are usually selenate (+6), selenite (+4), and
organo-selenium (-2). The presence of selenium in different oxidation states complicates some analytical
methods (Presser and Ohlendorf 1987). EPA recommends using standard reference samples to check for
the percentage recovery of each species of selenium (selenate, selenite and organo-selenium) during
initial testing of selenium methodologies for quality control and assurance.
If water samples are not filtered, particulate species such as elemental selenium and particulate organo-
selenium will also be measured. In addition, federal regulations at 40 CFR §122.45(c) generally requires
considering total recoverable metals when establishing effluent limits and reporting requirements.
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ANALYTICAL METHODS RECOMMENDED FOR MEASURING SELENIUM
IN WATER
EPA has several approved analytical methods under 40 CFR § 136 specifically for measuring total
selenium in water. These regulations state that measurements for NPDES permit applications and
permittee reporting should be made using analytical methods approved by EPA. Because EPA has
approved methods for analyzing selenium in water, these methods must be used for NPDES permits (40
CFR§ 122.21(g)(7), 122.41(j), 136.1, 136.3, and 136.6).
A complete list of EPA-approved analytical methods for selenium can be found at:
http://www.epa.gov/waterscience/methods/method/. Three EPA-approved methods that may be
sufficiently sensitive2 for the purposes of implementing a selenium water quality criterion are listed below
(Table L-l).
Table L-l. Suggested EPA-Approved Methods for Selenium in Water
Method
American Public Health Standard
Method 3 1 14 B (2009) or 3 1 14 C
(2009)
EPA Method 200.8, Rev 5.4
(1998)
EPA Method 200.9, Rev.2.2
(1994)
Technique
Hydride generation atomic absorption
spectrometry (HG-AAS)
Inductively coupled plasma mass
spectrometry (ICP-MS)
Stabilized temperature graphite
furnace atomic absorption (STGF-AA)
Method
detection limit
2^ig/L
7.9 Mg/L
0.6 ng/L
American Public Health Standard Method 3114 B
For measuring selenium in water, American Public Health Standard Method 3114 B uses the HG-AAS
technique. Method 3114 B has a method detection limit (MDL) of 2 (ig/L. Samples for dissolved analytes
should be filtered on-site through 0.45-micron capsule filters certified free of trace-element contamination
or other appropriate filtering equipment (Wilde et al. 1999). Dissolved samples should be preserved with
high purity hydrochloric acid or nitric acid to a pH less than 2.
For more information on choosing a sufficiently sensitive method, see the memorandum Analytical Methods for Mercury in
National Pollutant Discharge Elimination System (NPDES) Permits from James A. Hanlon, Director of the Office of Wastewater
Management, dated August 23, 2007, available at http://www.epa.gov/npdes/pubs/mercurymemo analyticalmethods.pdf
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For measuring total selenium, samples should not be filtered. In addition, all selenium in the sample
should be in the form of selenite (+4). Thus, the following pre-treatment steps to convert all selenium in
the sample to selenite are critical when using the HG-AAS method:
1. Boiling with persulfate to oxidize and digest organic material.
2. Boiling with hydrochloric acid to reduce selenate species to selenite.
3. Reduction by sodium borohydride to hydrogen selenide in the quartz tube of the AAS.
Optimal conversion conditions are essential for accurate results because too mild a reduction could lead to
incomplete reduction of selenate and too rigorous a reduction could lead to plating out of elemental
selenium (Cutter 1987, 1983; Presser and Barnes 1984, 1985).
Method 3114 B has the advantage that it is a fully validated method, is commonly used by many
laboratories, is relatively inexpensive, is less susceptible to background interference (Cutter 1987, 1983;
Presser and Barnes 1984, 1985), and has sufficient sensitivity to accurately measure what can be expected
in many lotic aquatic systems. However, this method may not be sufficiently sensitive for some lentic
aquatic systems where relatively lower selenium concentrations may need to be measured. If no selenium
is detected in a lentic system using this method, EPA recommends using a more sensitive analytical
method.
EPA Method200.8
EPA method 200.8 has a MDL of 7.9 (ig/L using the ICP-MS analytical technique. This method has the
advantage that no pre-treatment steps are necessary. However, this method may not be sufficiently
sensitive in many applications of the selenium criterion (Lamothe et al. 1999). If no selenium is detected
using this method, EPA recommends monitoring with a more sensitive method.
EPA Method200.9
Method 200.9 has a MDL of 0.6 (ig/L using the STGF-AA analytical technique. This method has the
advantage that it can detect selenium at very low concentrations. However, graphite furnace techniques
require careful matrix matching.
Of these three EPA approved methods, Method 3114 B using the HG-AAS technique is the most cost-
effective, with sufficient sensitivity and relatively low risk of interference in most cases. EPA Method
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200.8 may be used where appropriate, such as when selenium concentrations in effluent are known to be
higher than 7.9 (ig/L. EPA Method 200.9 may be used if a very low MDL is needed.
Some additional methods not approved by EPA that states and tribes might consider are:
• Collision/Reaction Cell Inductively Coupled Plasma Mass Spectroscopy (cICP-MS) (Garbarino
et al. 2005) - A relatively new technique that is a sensitive and selective detector for metal
analysis. However, isobaric interference can cause problems for quantitative determination as
well as identification based on the analyte pattern. Collision cells, reaction cells or other
interfaces reducing sample matrix effects that might otherwise interfere in the mass selective
determinative step are allowed in CWA analyses provided the method performance specifications
relevant to ICP-MS measurements are met
• Fluorometric Analysis,- a wet chemistry technique using diaminonapthalene. This method also
achieves acceptable precision and accuracy on standard reference samples (Olson 1969; Olson et
al. 1975; American Public Health Association Standard Method 3500, on-line version).
Methods for measuring different species of selenium dissolved in water are also available. These methods
determine the species of dissolved selenium present in a sample through differential digestion and hydride
generation atomic adsorption spectrophotometry (Cutter 1978, 1983; Presser and Barnes, 1984; 1985;
May et al. 2007). Selenite can be measured in samples with no pre-treatment. Selenate plus selenite can
be measured in samples subjected to boiling with hydrochloric acid. Subtraction of the measured selenite
fraction from the measured combined fraction would yield a measure of the selenate fraction. If a sample
is analyzed to measure total dissolved selenium as described above, then measurements of the combined
fraction can be subtracted to yield measurements of the dissolved organo-selenium fraction.
ANALYTICAL METHODS AVAILABLE FOR MEASURING SELENIUM IN
FISH TISSUE
EPA does not have approved methods under 40 CFR § 136 for measuring selenium in fish tissue.
However, states and tribes are not required to use EPA-approved methods for monitoring and assessment
of criteria attainment or other activities not related to permit applications or reports.
The techniques described above for analyzing selenium in water (HG-AAS, ICP-MS, and STGF-AA) can
be used to measure selenium in fish tissue if the samples are made soluble. Tissue samples are
homogenized and digested prior to analysis using strong acid or dry-ashing digestion as described below.
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A review of sample digestion techniques has been published (Ihnat 1992). Standard reference materials,
analytical duplicates, and matrix spike samples are recommended to determine the applicability of a
selected digestion procedure.
Strong acid digestion
Solid samples can be subjected to strong acid digestion to break down mineral and organic matrices.
Samples are typically dried and homogenized before digestion. Determination of percent moisture may be
part of the drying procedure. Note that some strong acid digestion methods may not be suitable for fish
tissue. Strong acid digestion methods are categorized by the type of material or amount of organic
material present (e.g., solid waste; biological tissue, plants, soil, sediment, rock, coal) and degrees of
tissue solubilization needed (extraction, leachate, or complete destruction). Methods differ in acid mixture
and degree and type of heating (EPA Method 3050B, Revision 2, 1996; EPA Method 200.2, Revision 2.8,
1994; Briggs and Crock, 1986; Taggart, 2002, chapters I, J, and K). High boiling acids (perchloric and
sulfuric) may lead to a loss of selenium if solutions are heated to dryness.
Dry-ashing digestion
Dry-ashing digestion is applicable to biological samples (Brumbaugh and Walther, 1989; May et al.,
2007). Biological samples are normally lyophilized (freeze-dried) and homogenized before digestion.
Determination of percent moisture may be part of the drying procedure. Dried solid samples are:
1. Boiled in nitric acid for solubilization and oxidation
2. Ashed at 500° C with magnesium nitrate to complete oxidation and decompose remaining organic
material
3. Heated with hydrochloric acid to dissolve the ash and reduce selenium to the selenite (+4) state
required for detection by HG-AAS.
Analytical methods available for measuring selenium in paniculate material
There are no 40 CFR § 136 methods for analyzing selenium in particulate material. However, states and
tribes are not required to use EPA-approved methods for monitoring and assessment of criteria attainment
or other activities not related to permit applications or reports.
The techniques described above for analyzing selenium in water (HG-AAS, ICP-MS, and STGF-AA) can
be used to measure selenium in particulate material after the sample has been separated from the water
and pre-treated using the same methods used for fish tissue. In order to obtain a particulate material
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sample, a water column sample should be filtered to separate the particulate material and bed sediment.
Various techniques for collection of suspended particulate material using filtration are available from the
EPA (e.g. Method 1669) and the U.S. Geological Survey (Moulton et al. 2002; USGS, Britton and
Greeson 1987). These techniques include:
• EPA Method 1669 (1996) includes filtration through a 0.45 (im capsule filter at the field site.
• USGS protocols for collection of phytoplankton and seston in rivers and streams as part of their
National Water Quality Assessment Program for watershed and habitat assessment
(http ://water.usgs .gov/nawqa/protocols .html).
• Textbooks such as Limnological Analyses address sampling of lakes using traditional techniques
including phytoplankton nets. (Wetzel and Likens 1991).
• Sampling of suspended material from estuaries where particulates are a substantial part of the
ecosystem is described in Doblin et al. (2005) as part of their work on the San Francisco Bay-
Delta Estuary.
• Separating suspended sediment using high-speed centrifugation and decantation when the
concentration of particulate material is relatively low (Horowitz et al. 1989).
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APPENDIX M: ABBREVIATIONS
Do not distribute, quote, or cite M-l Draft Document
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REFERENCE AND SITE ABBREVIATIONS
Reference
Bi:
Birkner 1978
Bu91:
Butler etal. 1991
Bu93:
Butler etal. 1993
Site
22
27
23
20
7
22
23
30
3
27
23
4
4
4
4
4
4
SP2
N2
SP2
N2
N2
N2
SP2
SP2
Miller's Lake, Wellington CO
Sweltzer Lake, Delta CO
Twin Butter Reservoir, Laramie WY
East Allen Reservoir, Medicine Bow WY
Galett Lake, Laramie WY
Miller's Lake, Wellington CO
Twin Butter Reservoir, Laramie WY
Larimer Highway 9 Pond, Fort Collins CO
Meeboer Lake, Laramie WY
Sweltzer Lake, Delta CO
Twin Butter Reservoir, Laramie WY
Uncompahgre River at Colona
Uncompahgre River at Colona
Uncompahgre River at Colona
Uncompahgre River at Colona
Uncompahgre River at Colona
Uncompahgre River at Colona
Spring Creek at La Boca
Navajo Reservoir, Piedra R. Arm, near La Boca
Spring Creek at La Boca
Navajo Reservoir, Piedra R. Arm, near La Boca
Navajo Reservoir, Piedra R. Arm, near La Boca
Navajo Reservoir, Piedra R. Arm, near La Boca
Spring Creek at La Boca
Spring Creek at La Boca
Species
FM
FM
FM
ID
ID
ID
ID
NPK
NPK
NPK
NPK
BhS
BnT
FS
MS
RT
WS
BhS
BT
BT
BB
ChC
cc
FM
SD
Fathead minnow
Fathead minnow
Fathead minnow
Iowa darter
Iowa darter
Iowa darter
Iowa darter
Northern plains killfish
Northern plains killfish
Northern plains killfish
Northern plains killfish
Bluehead sucker
Brown trout
Flannelmouth sucker
Mottled sculpin
Rainbow trout
White sucker
Bluehead sucker
Brown trout
Brown trout
Black bullhead
Channel catfish
Common carp
Fathead minnow
Speckled dace
Do not distribute, quote, or cite
M-2
Draft Document
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Reference
Bu95:
Butler etal. 1995
Site
SP2
ME2
ME3
NW
SJ1
SJ3
ME3
SJ1
SJ3
ME4
ME3
SJ1
SJ3
HD2
ME1
ME2
ME4
ME3
we
SJ1
HD2
ME2
ME4
ME3
SJ3
ME3
ME4
ME3
SJ1
ME1
Spring Creek at La Boca
McElmo Cr., downstream from Alkali Canyon
McElmo Cr., upstream from Yellow Jacket Canyon
Navajo Wash near Towaoc
San Juan R. at Four Corners
San Juan R. at Mexican Hat Utah
McElmo Cr., upstream from Yellow Jacket Canyon
San Juan R. at Four Corners
San Juan R. at Mexican Hat Utah
McElmo Cr., downstream from Yellow Jacket Canyon
McElmo Cr., upstream from Yellow Jacket Canyon
San Juan R. at Four Corners
San Juan R. at Mexican Hat Utah
Hartman Draw near mouth, at Cortez
McElmo Cr. at Hwy. 160, near Cortez
McElmo Cr., downstream from Alkali Canyon
McElmo Cr., downstream from Yellow Jacket Canyon
McElmo Cr., upstream from Yellow Jacket Canyon
Woods Canyon near Yellow Jacket
San Juan R. at Four Corners
Hartman Draw near mouth, at Cortez
McElmo Cr., downstream from Alkali Canyon
McElmo Cr., downstream from Yellow Jacket Canyon
McElmo Cr., upstream from Yellow Jacket Canyon
San Juan R. at Mexican Hat Utah
McElmo Cr., upstream from Yellow Jacket Canyon
McElmo Cr., downstream from Yellow Jacket Canyon
McElmo Cr., upstream from Yellow Jacket Canyon
San Juan R. at Four Corners
McElmo Cr. at Hwy. 160, near Cortez
Species
WS
BhS
BhS
BhS
BhS
BhS
BB
ChC
ChC
cc
cc
cc
cc
FM
FM
FM
FM
FM
FM
FS
FS
FS
FS
FS
FS
GnS
RSh
RSh
RSh
SD
White sucker
Bluehead sucker
Bluehead sucker
Bluehead sucker
Bluehead sucker
Bluehead sucker
Black bullhead
Channel catfish
Channel catfish
Common carp
Common carp
Common carp
Common carp
Fathead minnow
Fathead minnow
Fathead minnow
Fathead minnow
Fathead minnow
Fathead minnow
Flannelmouth sucker
Flannelmouth sucker
Flannelmouth sucker
Flannelmouth sucker
Flannelmouth sucker
Flannelmouth sucker
Green sunfish
Red sunfish
Red sunfish
Red sunfish
Speckled dace
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M-3
Draft Document
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Reference
Bu97:
Butler etal. 1997
Ca:
Casey and
Fo:
Formation 2012
Site
ME2
ME3
NW
SJ1
HD2
MUD2
MNP2
MUD2
WCP
CHI
MUD2
MNP3
DC
LC
CC-1A
CC-3A
CC-150
CC-350
CC-75
DC
HS
HS-3
LSV-2C
LSV-4
SFTC
CC-1A
CC-3A
CC-150
McElmo Cr., downstream from Alkali Canyon
McElmo Cr., upstream from Yellow Jacket Canyon
Navajo Wash near Towaoc
San Juan R. at Four Corners
Hartman Draw near mouth, at Cortez
Mud Cr. at Hwy. 32, near Cortez
Large pond south of G Road, southern Mancos Valley
Mud Cr. at Hwy. 32, near Cortez
Pond on Woods Canyon at 15 Road
Cahone Canyon at Hwy. 666
Mud Cr. at Hwy. 32, near Cortez
Pond downstream from site MNP2, southern Mancos Valley
Deerlick Creek
Luscar Creek
Crow Creek - 1A
Crow Creek - 3A
Crow Creek- 150
Crow Creek - 350
Crow Creek - 75
Deer Creek
Hoopes Spring
Hoopes Spring - 3
Sage Creek - 2C
Sage Creek - 4
South Fork Tincup Creek
Crow Creek - 1A
Crow Creek - 3A
Crow Creek- 150
Species
SD
SD
SD
SD
Su
BhS
FM
FM
FM
GnS
GnS
SB
RT
RT
BnT
BnT
BnT
BnT
BnT
BnT
BnT
BnT
BnT
BnT
BnT
Sc
Sc
Sc
Speckled dace
Speckled dace
Speckled dace
Speckled dace
Sucker
Bluehead sucker
Fathead minnow
Fathead minnow
Fathead minnow
Green sunfish
Green sunfish
Smallmouth bass
Rainbow trout
Rainbow trout
Brown trout
Brown trout
Brown trout
Brown trout
Brown trout
Brown trout
Brown trout
Brown trout
Brown trout
Brown trout
Brown trout
Sculpin
Sculpin
Sculpin
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M-4
Draft Document
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Reference
Gr:
Grassoetal. 1995
HB:
Hamilton and
Buhl 2004
Le:
Lemly 1985
Site
CC-350
CC-75
DC
HS
HS-3
LSV-2C
LSV-4
SFTC
17
17
LEMC
BA
BE
HR
BA
BE
HR
BA
BE
HR
BA
BE
HR
BA
BE
HR
Crow Creek - 350
Crow Creek - 75
Deer Creek
Hoopes Spring
Hoopes Spring - 3
Sage Creek - 2C
Sage Creek - 4
South Fork Tincup Creek
Arapahoe Wetlands Pond
Arapahoe Wetlands Pond
Lower East Mill Creek
Badin Lake
Belews Lake
High Rock Lake
Badin Lake
Belews Lake
High Rock Lake
Badin Lake
Belews Lake
High Rock Lake
Badin Lake
Belews Lake
High Rock Lake
Badin Lake
Belews Lake
High Rock Lake
Species
Sc
Sc
Sc
Sc
Sc
Sc
Sc
Sc
FM
WS
CT
BB
BB
BB
CC
CC
CC
FM
FM
FM
GnS
GnS
GnS
WM
WM
WM
Sculpin
Sculpin
Sculpin
Sculpin
Sculpin
Sculpin
Sculpin
Sculpin
Fathead minnow
White sucker
Cutthroat trout
Black bullhead
Black bullhead
Black bullhead
Common carp
Common carp
Common carp
Fathead minnow
Fathead minnow
Fathead minnow
Green sunfish
Green sunfish
Green sunfish
Western mosquitofish
Western mosquitofish
Western mosquitofish
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M-5
Draft Document
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Reference
Sa87:
Saiki and
Lowe 1987
Sa93:
Saiki etal. 1993
St:
Stephens et al. 1988
Site
BA
BE
HR
KP11
KP2
KP8
SLD
VP26
VW
GT4
GTS
SJR2
SJR3
GT4
GTS
SJR2
SJR3
GT4
GTS
SJR2
SJR3
M4720
M4720
Badin Lake
Belews Lake
High Rock Lake
Kesterson Pond 1 1
Kesterson Pond 2
Kesterson Pond 8
San Luis Drain
Volta Pond 26
Volta Wasteway
Salt Slough at San Luis Wildlife Refuge
Mud Slough at San Luis Wildlife Refuge
San Joaquin R. above Hills Ferry Rd.
San Joaquin R. at Durham Ferry Recreation Area
Salt Slough at San Luis Wildlife Refuge
Mud Slough at San Luis Wildlife Refuge
San Joaquin R. above Hills Ferry Rd.
San Joaquin R. at Durham Ferry Recreation Area
Salt Slough at San Luis Wildlife Refuge
Mud Slough at San Luis Wildlife Refuge
San Joaquin R. above Hills Ferry Rd.
San Joaquin R. at Durham Ferry Recreation Area
Marsh 4720
Marsh 4720
Species
RSh
RSh
RSh
WM
WM
WM
WM
WM
WM
Bg
Bg
Bg
Bg
LMB
LMB
LMB
LMB
WM
WM
WM
WM
BB
CC
Red shiner
Red shiner
Red shiner
Western mosquitofish
Western mosquitofish
Western mosquitofish
Western mosquitofish
Western mosquitofish
Western mosquitofish
Bluegill
Bluegill
Bluegill
Bluegill
Largemouth bass
Largemouth bass
Largemouth bass
Largemouth bass
Western mosquitofish
Western mosquitofish
Western mosquitofish
Western mosquitofish
Black bullhead
Common carp
Do not distribute, quote, or cite
M-6
Draft Document
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