United States

Environmental Protection

Agency

Office of Water

4304

EPA-822-D-04-001

November 2004

Draft

Aquatic Life

Water Quality Criteria

for Selenium

- 2004
Draft November 12, 2004 

Draft

Aquatic Life Water Quality Criteria for

Selenium

2004

November 2004

U.S. Environmental Protection Agency

Office of Water

Office of Science And Technology

Washington, D.C.
ii Draft November 12, 2004 

NOTICES

This document has been reviewed by the Health and Ecological Effects Criteria Division, Office

of Science and Technology, U.S. Environmental Protection Agency, and approved for

publication.

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
iii Draft November 12, 2004 

ACKNOWLEDGMENTS

Dennis O. McIntyre Larry T. Brooke

Tyler K. Linton University of Wisconsin-Superior

William H. Clement Superior, Wisconsin

Gregory J. Smith

Manoel Pacheco

Great Lakes Environmental Center

Columbus, Ohio

Charles Delos 

(document coordinator)

USEPA 

Health and Ecological Effects Criteria Division

Washington, D.C.
iv Draft November 12, 2004 

Table of Contents

Introduction ........................................................................ 1

Selenium Chemistry ........................................................... 1

Inorganic Selenium ............................................................ 2

Organoselenium .............................................................. 4

Departure from Thermodynamic Equilibrium ....................................... 5

Physical Distribution of Species in Surface Water .................................... 5

Sources of Selenium to Aquatic Systems ........................................... 6

Selenium Biogeochemistry ...................................................... 7

Narrow Margin Between Sufficiency and Toxicity ................................... 8

Selenium Document Information ................................................ 10

Acute Toxicity of Selenite ............................................................ 12

Acute Toxicity of Se(IV) to Freshwater Animals .................................... 12

Hyalella (amphipod) ................................................ 12

Ceriodaphnia (cladoceran) .............................................. 12

Daphnia (cladoceran) .................................................. 13

Hydra ............................................................... 13

Morone (striped bass) .................................................. 13

Pimephales (fathead minnow) ............................................ 13

Gammarus (amphipod) ................................................. 13

Jordanella (flagfish) ................................................... 13

Oncorhynchus (salmonid) ............................................... 14

Lepomis (bluegill) ..................................................... 14

Se(IV) Freshwater Final Acute Value Determination ................................ 14

Acute Toxicity of Se(IV) to Saltwater Animals ..................................... 15

Argopecten (bay scallop) ................................................ 15

Melanogrammus (haddock) .............................................. 16

Cancer (dungeness crab) ................................................ 16

Penaeus (brown shrimp) ................................................ 16

Acartia (copepod) ..................................................... 16

Americamysis (Mysidopsis) (mysid) ....................................... 16

Spisula (surf clam) ..................................................... 16

Morone (striped bass) .................................................. 17

Paralichthys (summer flounder) .......................................... 17

Callinectes (blue crab) .................................................. 17

Crassostrea (Pacific oyster) ............................................. 17

Mytilus (blue mussel) ................................................... 17

Pseudopleuronectes (winter flounder) ...................................... 17

Se(IV) Saltwater Final Acute Value Determination ................................. 18

Acute Toxicity of Selenate ........................................................... 19

Sulfate-dependent Toxicity of Selenate ........................................... 19

Sulfate Correction ............................................................ 19

Acute Toxicity of Se(VI) to Freshwater Animals (Sulfate Adjusted Values) .............. 21
v Draft November 12, 2004 

Ceriodaphnia (cladoceran) .............................................. 21

Hyalella (amphipod) ................................................... 21

Daphnia (cladoceran) .................................................. 21

Gammarus (amphipod) ................................................. 22

Xyrauchen (razorback sucker) ............................................ 22

Gila (bonytail) ........................................................ 22

Pimephales (fathead minnow) ............................................ 23

Ptychocheilus (Colorado squawfish) ....................................... 23

Oncorhynchus (salmonid) ............................................... 23

Lepomis (bluegill) ..................................................... 24

Ictalurus (channel catfish) ............................................... 24

Se(VI) Freshwater Final Acute Value Determination ................................ 24

Acute Toxicity of Se(VI) to Saltwater Animals ..................................... 24

Se(VI) Saltwater Final Acute Value Determination .................................. 24

Comparison of Selenite and Selenate Acute Toxicity ................................ 25

Review and Analysis of Chronic Data ................................................... 55

Selection of Medium for Expressing Chronic Criterion ............................... 55

Calculation of Chronic Values .................................................. 58

Evaluation of Freshwater Chronic Data for Each Species ............................. 60

Brachionus calyciflorus (freshwater rotifer) ................................ 61

Oncorhynchus tshawytscha (chinook salmon) ............................... 61

Oncorhynchus mykiss (rainbow trout) ...................................... 62

Oncorhynchus clarki (cutthroat trout) ...................................... 64

Salvelinus fontinalis (brook trout) ........................................ 64

Pimephales promelas (fathead minnows) ................................... 65

Catostomus latipinnis (flannelmouth sucker) ................................ 68

Xyrauchen texanus (razorback sucker) ..................................... 68

Lepomis macrochirus (bluegill sunfish) .................................... 69

Morone saxitilis (Striped bass) ........................................... 73

Formulation of the Final Chronic Value (FCV) for Selenium .......................... 74

National Criteria ................................................................... 82

Implementation .................................................................... 82

Appendices

A. Information Used in the Sulfate Correction of Selenate Acute Toxicity ................. A-1

B. Toxicity of Selenium to Aquatic Plants .......................................... .B-1

C. Bioconcentration and Bioaccumulation of Selenium ................................ C-1

D. Environmental Factors Affecting Selenium Toxicity and Bioaccumulation .............. D-1

E. Site-specific Considerations ................................................... E-1

F. Other Data .................................................................. F-1

G. Unused Data ............................................................... G-1

H. Data Used in Regression Analysis of Selenium in Whole-body Fish Tissue with Selenium in

Muscle, Ovary and Liver Tissue ................................................ H-1
vi Draft November 12, 2004 

I. Summaries of Chronic Studies Considered for FCV Derivation ........................ I-1

J. Selenium in Fish Samples Collected From 112 Sites as Part of U.S. Fish and Wildlife’s National

Biomonitoring Program, 1979-1981 .............................................. J-1

List of Tables

Table A. Particulate and dissolved selenium .............................................. 7

Table 1a. Acute Toxicity of Selenium to Freshwater Animals ............................... 26

Table 1b. Acute Toxicity of Selenium to Saltwater Animals ................................. 40

Table 2a. Ranked Freshwater Genus Mean Acute Values .................................. 43

Table 2b. Ranked Saltwater Genus Mean Acute Values .................................... 47

Table 3a. Ratios of Freshwater Species Mean Acute Values for Se ........................... 49

Table 3b. Ratios of Saltwater Species Mean Acute Values for Sel ............................ 51

Table 4. Freshwater Chronic Values from Acceptable Tests ............................... 78

List of Figures

Figure 1. Ranked summary of selenite GMAVs (freshwater). ................................ 52

Figure 2. Ranked summary of selenate GMAVs (saltwater). ................................. 53

Figure 3. Ranked summary of selenate GMAVs (freshwater) at a sulfate level of 100 mg/L. ....... 54

Figure 4. The quantile regression curves project median selenium concentrations in the whole 

body of bluegill, largemouth bass, tilapia and carp as a function of selenium 

concentrations in their tissues. ................................................. 57

Figure 5. Reductions in survival, growth or other responses of organisms are often modeled 

as a sigmoid function of increasing concentrations of selenium, or any other toxic 

substance. ................................................................. 59
1 Draft November 12, 2004 

Introduction

This document provides 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 standards to protect designated uses. While this

document constitutes the U.S. Environmental Protection Agency’s (U.S. EPA) scientific

recommendations regarding ambient concentrations of selenium, this document does not substitute for

the CWA or U.S. EPA’s regulations; nor is it a regulation itself. Thus, it cannot impose legally binding

requirements on the U.S. EPA, States, Tribes or the regulated community, and might not apply to a

particular situation based upon the circumstances. Interested parties are free to raise questions and

objections about the substance of this guidance and the appropriateness of the application of this

guidance to a particular situation. State and Tribal decision-makers retain the discretion to adopt

approaches on a case-by-case basis that differ from this guidance when appropriate. The U.S. EPA may

change this guidance in the future.

This document establishes water quality criteria for protection of aquatic life for selenium. Under

Section 304(a) of the CWA, U.S. EPA is to periodically revise water quality criteria to accurately reflect

the latest scientific knowledge. Toward this end, a U.S. EPA-sponsored Peer Consultation Workshop on

Selenium Aquatic Toxicity and Bioaccumulation on May 27-28, 1998 brought together experts in

selenium research to discuss issues related to the chronic criterion for selenium. As a result of findings

from the workshop and the fact that a substantial body of literature on the chronic toxicity of selenium

has accumulated since the 1987 document was published, U.S. EPA has decided to update the acute and

chronic criteria for selenium.

The criteria presented herein supersede all previous national aquatic life water quality criteria for

selenium (U.S. EPA 1976, 1980a, 1987a, 1995). 

Selenium Chemistry

Water quality criteria are being derived for total selenium measured as selenite-Se plus selenate-Se, but a

variety of forms of selenium can occur in water and tissue. Selenium in aquatic ecosystems exists in a

broad range of oxidation states: (+ VI) in selenates (HSeO4

-

, SeO4

2-) and selenic acid (H2SeO4

), (+ IV) in

selenites (HSeO3

-

, SeO3

2-) 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
2 Draft November 12, 2004 

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, SeO3

2-, HSeO3

-

, and SeO4

2- 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 [E

0

(SeO4

2-/H2SeO3

) = 1.15 V;

E

0

(Cr2O7

2-/Cr3+) = 1.33V; E0

(SO4

2-/H2SO3

) = 0.200V (standard potentials in acid solution: Weast 1969)],

whereas selenide is a much stronger reducing agent than sulfide [E

0

(Se/H2Se) = -0.36 V; E

0

[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 (SeO4

2-) exhibits considerable kinetic stability in the presence

of reducing agents (Cotton and Wilkinson 1988). The radius of SeO4

2- is comparable to that of SO4

2-

(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), crustacea

(Olge 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 relationship was shown to

exist between acute selenate toxicity to aquatic organisms and ambient sulfate concentrations (Brix et al.

2001a ). 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 et al. 1989).

Selenous acid species (HSeO3

-

 and SeO3

2-) 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.
3 Draft November 12, 2004 

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 × 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 SeO3

2- 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 Ruppia maritima (Bailey et al. 1995), and

specific uptake systems have been demonstrated in some 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
4 Draft November 12, 2004 

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

substantially at the lower pH values. Differences in speciation are suggested to be the cause of these

differences. Selenate exists as the divalent ion SeO4

2- 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 Macoma balthica has been reported (Luoma et al. 1992). The

bioavailability of elemental selenium to M. 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 preceeded 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 Se0

/H2Se couple

falls even below the H+

/H2

 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
5 Draft November 12, 2004 

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 μg/L) to 53.7% (1.02

μg/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 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. 1992). 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 COH)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 particulate phase (0.45μm 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% (Text Table
6 Draft November 12, 2004 

A). 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.

Up 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

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.

Sources of Selenium to Aquatic Systems

Selenium occurs in many soil types and enters ground and surface waters through natural weathering

process such as erosion, leaching and runoff. The national average concentration of selenium in nonseleniferous surface water ranges from 0.1 to 0.4 μg Se/L (Maier and Knight 1993). Elevated levels of

selenium occur in surface waters when substantial quantities of selenium enter surface waters from both

natural and anthropogenic sources. It is abundant in the alkaline soils of North America from the Great

Plains. 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. 

Ecological impacts have been observed where selenium is concentrated through irrigation practices in

areas with seleniferous soils. Selenium also occurs in sulfide deposits of copper, lead, mercury, silver

and zinc and can be released during the mining and smelting of these ores. In addition, selenium occurs

naturally in coal and fuel oil and is emitted in flue gas and in fly ash during combustion. Some selenium

then enters surface waters in drainage from fly-ash ponds and in runoff from fly-ash deposits on land. 

Notable examples of systems that have been affected by selenium originating from coal ash include
7 Draft November 12, 2004 

Belews Lake, NC, where 16 of the 20 species originally present were eliminated within a few years after

discharge began, and Hyco Reservoir, NC, where selenium toxicity was associated with fish larval

mortality (Gillespie and Baumann 1986).

Text Table A. Particulate and dissolved selenium as a function of total selenium in freshwater and

marine aquatic ecoystems.

Reference Waterbody

Particulate Se

(% of Total)

Fraction

dissolved, fd

Cutter 1989 Carquinezitist, CA 20 - 40 0.6 - 0.8

Cutter 1986 Hyco Reservoir, NC 0 1

Tanizaki et al. 1992 Japanese Rivers 16 0.84

Luoma et al. 1992 San Francisco Bay, CA 22 - 31 0.69-0.78

Cumbie and VanHorn, 1978 Belews Lake, NC 8 0.92

GLEC 1997 Unnamed Stream, Albright, WV 4 0.96

Wang et al. 1995 Finnish Lakes 10 0.9

Lahermo et al. 1998 Finnish Streams 8 0.92

Hamilton et al. 2001ab Adobe Creek, Fruita, CO 18 0.82

Hamilton et al. 2001a,b North Pond, Fruita, CO 0 1

Hamilton et al. 2001a,b Fish Ponds, Fruita, CO 7 0.93

Nakamoto and Hassler 1992 Merced River, CA 0 1

Nakamoto and Hassler 1992 Salt Slough, CA 4 0.96

Welsh and Maughan 1994 Cibola Lake, CA 39 0.62

Welsh and Maughan 1994 Hart Mine Marsh, Blythe, CA 6 0.94

Welsh and Maughan 1994 Colorado River, Blythe, CA 11 0.89

Welsh and Maughan 1994 Palo Verda Oxbow Lake, CA 33 0.67

Welsh and Maughan 1994 Palo Verda Oufall Drain, CA 0 1

Welsh and Maughan 1994 Pretty Water Lake, CA 21 0.79

Selenium Biogeochemistry

The current understanding of the biogeochemistry of selenium has recently been reviewed by Fan et al.

(2002). Their review clearly shows the extreme complexity of selenium biogeochemistry in aquatic

environments. Fan et al. describe the selenium biogeochemical cycle as follows: dissolved selenium

oxyanions are primarily absorbed by aquatic producers, including microphytes and bacteria, and

biotransformed into organoselenium form(s) and selenium element (SeO

). These, together with other

particle-bound selenium sources, constitute the particulate selenium fraction of the water column, and

they are poorly understood (Zawislanski and McGrath, 1998). Once accumulated in the aquatic primary

and secondary producers, selenium can be transferred through various aquatic consumers (e.g.
8 Draft November 12, 2004 

zooplankton, insect larvae, larval fish, bivalves) into the top predators, including aquatic birds and

piscivorous fish. Selenium can be further chemically transformed through the food chain transfer process.

The microscopic planktonic organisms, including microphytes (cyanobacteria and phytoplankton),

bacteria, protozoa, and zooplankton are major components of the particulate matter in the water column.

The particulate matter, in turn, forms the basis for detrital materials which can settle onto the sediment,

and become the food source for sediment organisms. Suspended particulate matter can also be

mineralized in the water column. In addition to this selenium input into the sediment, waterborne selenite

and selenate can be physically adsorbed onto the sediment particles, ingested, absorbed, and transformed

by the sediment organisms. Sediment-bound selenate and selenite can be reduced to insoluble Se0 by

anaerobic microbial activities. This and water column-derived Se0 can be reduced further to inorganic

and organic selenides (-II form), and/or reoxidized to selenite and selenate by microorganisms in the

sediment and/or in the digestive tracts of sediment macroinvertebrates. Selenides can enter the food chain

via absorption and/or ingestion (by chironomids or tubificid worms, for example) into sediment

organisms, or be oxidized to selenite and selenate. Selenium of different oxidation states can be further

biotransformed by sediment organisms and transferred up the food chain. Selenium biotransformation,

bioaccumulation, and transfer through both sediment and water column foodwebs constitute the major

biogeochemical pathways in aquatic ecosystems.

In addition to accumulating selenium into the biomass, the aquatic producers are the main factors

controlling the volatilization of selenium via the production of methylated selenides including,

dimethylselenide (DMSe) and dimethyldiselenide (DMDSe). These methylated selenides can be oxidized

to selenite, or can exit the water column into the atmosphere. Selenium volatilization into the atmosphere

may represent an important process responsible for significant loss of selenium in some aquatic systems.

Methylated selenides can also be generated from dissolved selenonium precursor(s) released by aquatic

producers into the water. Moreover, other organoselenium forms can be released into the water by

aquatic producers, and are reoxidized to selenite and/or reabsorbed by aquatic producers.

Narrow Margin Between Sufficiency and Toxicity

Of all the priority and non-priority pollutants, selenium has the narrowest range of what is beneficial for

biota and what is detrimental. Selenium is an essential element required as a mineral cofactor in the

biosynthesis of glutathione peroxidases. All of the classic glutathione peroxidases contain selenium and
9 Draft November 12, 2004 

are found to be involved in the catalytic reaction of these many enzymes (Allan 1999). The major

function of the glutathione peroxidases was found to involve the reduction of hydrogen peroxide to water

at the expense of the oxidation of glutathione, the enzyme’s cofactor. Aquatic and terrestrial organisms

require 0.5 μg/g dry weight (dw) of selenium in their diet to sustain metabolic processes, whereas

concentrations of selenium that are only an order of magnitude greater than the required level have been

shown to be toxic to fish. Selenium deficiency has been found to affect humans (U.S. EPA 1987a), sheep

and cattle (U.S. EPA 1987a), deer (Oliver et al. 1990) fish (Thorarinsson et al. 1994; Wang and Lovell

1997; Wilson et al. 1997; U.S. EPA 1987a), 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 1987a; 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 1987a; Wehr and Brown 1985). 

Selenium has been shown to mitigate the toxic effects of arsenic, cadmium, copper, inorganic and

organic mercury, silver, ofloxacin, methyl parathion and the herbicide paraquat to biota in both aquatic

and terrestrial environments (Bjerregaard 1988a, b; Cuvin and Furness 1988; Ding et al. 1988; Krizkova

et al. 1996; Malarvizhi and Usharani 1994; Micallef and Tyler 1987; Patel et al. 1988; Paulsson and

Lundbergh 1991; Pelletier 1986b, 1988; Phillips et al. 1987; Ramakrishna et al. 1988; Rouleau et al.

1992; Salte et al. 1988; Siegel et al. 1991; Szilagyi et al. 1993; U.S. EPA 1987a). Selenium pretreatment

resulted in reduced effects in 128-hr old, but not 6-hr old, embryos of Oryzias latipes from cadmium and

mercury, whereas prior exposure to selenium did not affect the sensitivity of white suckers to cadmium

(U.S. EPA 1987a). In contrast, Birge et al. and Huckabee and Griffith reported that selenium and

mercury acted synergistically in producing toxic effects to fish embryos (U.S. EPA 1987a). Selenium is

reported to reduce the uptake of mercury by some aquatic species (Southworth et al. 1994; U.S. EPA

1987a), to have no effect on uptake of mercury by a mussel, and to increase the uptake of mercury by

mammals and some fish (U.S. EPA 1987a). Selenium augmented accumulation of cadmium in some

tissues of the shore crab, Carcinus maenas (U.S. EPA 1987a). The available data do not show whether

the various inorganic and organic compounds and oxidation states of selenium are equally effective

sources of selenium as a trace nutrient, or as reducing the toxic effects of various pollutants.
10 Draft November 12, 2004 

Selenium Document Information

All concentrations reported herein are expressed as selenium, not as the chemical tested. Although

Se(VI) is expected to be the predominant oxidation state at chemical equilibrium in oxygenated alkaline

waters, the rate of conversion of Se(IV) to Se(VI) seems to be slow in most natural waters. Therefore, it

was assumed that when Se(IV) was introduced into stock or test solutions, it would persist as the

predominate state throughout the test, even if no analyses specific for the Se(IV) oxidation state were

performed. Similarly, it was assumed that when Se(VI) was introduced into stock or test solutions, it

would persist as the predominant state throughout the test, even if no analyses specific for Se(VI) were

performed.

An understanding of the "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 the

Guidelines, and the response to public comments (U.S. EPA 1985a) is helpful for understanding the

derivation of the acute criteria for selenium. Briefly, the Guidelines procedure involves the following

steps: (1) Acute toxicity test data is gathered from all suitably conducted studies. Data are to be

available for species in a minimum of eight families representing a diverse assemblage of taxa. (2) The

Final Acute Value (FAV) is derived by extrapolation or interpolation to a hypothetical genus more

sensitive than 95 percent of a diverse assemblage of taxa. The FAV, which represents an LC50 or EC50, is

divided by two in order to obtain an acute criterion protective of nearly all individuals in such a genus. 

(3) Chronic toxicity test data (longer-term survival, growth, or reproduction) are needed for at least three

taxa. Most often the chronic criterion is set by determining an appropriate acute-chronic ratio (the ratio

of acutely toxic concentrations to the chronically toxic concentrations) and applying that ratio to the FAV

from the previous step. (4) When necessary, the acute and/or chronic criterion may be lowered to protect

critically important species. 

The chronic criteria procedure explicitly set forth in the Guidelines (Step 3 above) is not well suited to

bioaccumulative contaminants for which diet is the primary route of aquatic life exposure. 

Consequently, that procedure was not used for deriving the chronic criterion for selenium either in the

original 1987 criteria document or in this update. Rather, to accord with other provisions of the

Guidelines, it was necessary to apply what the Guidelines refer to as “appropriate modifications” of the

procedures in order to obtain a criterion “consistent with sound scientific evidence”, as will be described

in a later section.
11 Draft November 12, 2004 

Results of such intermediate calculations as recalculated LC50 values and Species Mean Acute Values are

given to four significant figures to prevent roundoff error in subsequent calculations, not to reflect the

precision of the value. The latest comprehensive literature search for information for this document was

conducted in August 2001; some more recent information was included.

The body of this document contains only the information on acute and chronic toxicity of selenium that is

relevant to the derivation of the acute and chronic criteria. Supporting information on the toxicity and

bioaccumulation of selenium, and the data that were reviewed and not used in deriving the criteria are

provided in appendices and include: sulfate correction of selenate acute toxicity (Appendix A); toxicity

to aquatic plants (Appendix B); bioconcentration and bioaccumulation (Appendix C); environmental

factors affecting selenium toxicity and bioaccumulation (Appendix D); site-specific considerations

(Appendix E); other data (Appendix F); unused data (Appendix G); tissue relationships (Appendix H);

chronic data summaries (Appendix I); and background Se levels (Appendix J).
12 Draft November 12, 2004 

Acute Toxicity of Selenite

Data that may be used, according to the Guidelines, in the derivation of Final Acute Values for selenite

are presented in Tables 1a and 1b. The following text presents a brief overview of the acceptable data

obtained for selenite, followed by a discussion of the more sensitive, and commercially and

recreationally important species. A ranking of the relative sensitivity of selenite to selenate for each

genera is listed in Tables 2a and 2b.

Acute Toxicity of Se(IV) to Freshwater Animals

Acceptable data on the acute effects of selenite in freshwater are available for 14 species of invertebrates

and 20 species of fish (Table 1a). These 34 species satisfy the eight family provision specified in the

Guidelines. Invertebrates are both the most sensitive and the most tolerant freshwater species to selenite

with Species Mean Acute Values (SMAV) ranging from 440 μg/L for the crustacean, Ceriodaphnia

dubia, to 203,000 μg/L for the leech, Nephelopsis obscura. The selenite SMAVs for fishes range from

1,783 μg/L for the striped bass, Morone saxatilis, to 35,000 μg/L for the common carp, Cyprinus carpio. 

The following text presents a species-by-species discussion of the eight most sensitive genera, plus all

commercially and recreationally important species.

Hyalella (amphipod)

The most sensitive freshwater genus is the amphipod, Hyalella, with a Genus Mean Acute Value

(GMAV) of 461.4 μg Se/L. The GMAV is derived from five 96-hr acute flow-through measured tests

where the LC50 values ranged from 340 to 670 μg Se/L (GLEC 1998; Halter et al. 1980). A sixth test

conducted under non flow-through conditions is also listed in Table 1a (Brasher and Ogle 1993), but the

Guidelines recommend using flow-through measured data in preference to static or renewal data. 

Ceriodaphnia (cladoceran)

The second most sensitive freshwater genus is Ceriodaphnia, with a GMAV of <515.3 μg Se/L that is

derived from the geometric mean of the C. affinis (<603.6 μg Se/L) and C. dubia (440 μg Se/L) SMAVs. 

Four static unmeasured 48-hr studies are available for C. affinis where the LC50 values ranged from <480

to 720 μg Se/L (Owsley 1984; Owsley and McCauley 1986). The one available C. dubia acute study was

conducted by GLEC (1999) that exposed <24-hr old neonates to sodium selenite for 48 hours under flowthrough measured conditions. The resultant 48-hr LC50 value was 440 μg Se/L, which is the most

sensitive SMAV for selenite in the database. 
13 Draft November 12, 2004 

Daphnia (cladoceran)

The eleven available acute values are used to calculate the Daphnia magna SMAV of 905.3 μg Se/L

(acute LC50 values ranged from 215 to 3,020 μg Se/L), but only one flow-through measured acute LC50

test value of 1,987 μg Se/L is used for the for D. pulex SMAV (a second static measured test conducted

by Reading (1979) is listed, but not used to calculate the SMAV) . The resultant GMAV of 1,341 μg

Se/L for Daphnia is the third most sensitive for selenite. 

Hydra

The fourth most sensitive freshwater genus is Hydra, with a GMAV of 1,700 μg Se/L. The GMAV is

derived from the one available static-measured test conducted by Brooke et al. (1985).

Morone (striped bass)

Two 96-hr static unmeasured tests are available for the striped bass, Morone saxatilis, and the LC50

values were 1,325 and 2,400 μg Se/L (Palawski et al. 1985). The geometric mean of the two values yield

the GMAV of 1,783 μg Se/L.

Pimephales (fathead minnow)

A total of 16 fathead minnow acute studies are presented in Table 1a, but only the eight flow-through

measured LC50 values are used to derive the GMAV of 2,209 μg Se/L. The eight flow-through LC50

values ranged from 620 to 5,200 μg Se/L (Cardwell et al. 1976a,b; GLEC 1998; Kimball manuscript).

Gammarus (amphipod)

The seventh most sensitive freshwater genus is Gammarus, with a GMAV of 3,489 μg Se/L that is

derived from the geometric mean of five flow-through measured studies (GLEC 1998, 1999) where the

LC50 values ranged from 1,800 to 10,950 μg Se/L. Two static measured acute studies were conduced by

Brooke et al. (1985) and Brooke (1987), but as recommended by the Guidelines, were not used to

calculate the SMAV for this species.

Jordanella (flagfish)

The eighth most sensitive freshwater genus is Jordanella, with a GMAV of 6,500 μg Se/L. The GMAV

is derived from the one available 96-hr flow-through measured test conducted by Cardwell et al.

(1976a,b) that exposed Jordanella floridae to selenium dioxide.
14 Draft November 12, 2004 

Oncorhynchus (salmonid)

The GMAV of 10,580 μg Se/L for the commercially important salmonid Oncorhynchus is derived from

the geometric mean of the coho salmon (O. kisutch; 7,240 μg Se/L), chinook salmon (O. tshawytscha;

15,596 μg Se/L) and rainbow trout (O. mykiss; 10,488 μg Se/L) SMAVs. Three static unmeasured 96-hr

studies are used to calculate the coho salmon SMAV where the LC50 values ranged from 3,578 to 13,600

μg Se/L (Hamilton and Buhl 1990b; Buhl and Hamilton 1991). A fourth coho salmon LC50 value is

available for an acute test initiated with the tolerant alevin life stage (Buhl and Hamilton 1991), but

based on Guideline recommendations this value is not used when data are available from a more sensitive

life stage. 

Six acute chinook salmon static unmeasured 96-hr acute studies conducted with the more sensitive postalevin life stage of the fish are used to determine the 15,596 μg Se/L SMAV for the species and the LC50

values ranged from 8,150 to 23,400 μg Se/L (Hamilton and Buhl 1990b). The two acute studies

conducted with the tolerant eyed egg and alevin life stages by the same authors are not used in the

SMAV determination as recommended by the Guidelines. Hamilton and Buhl (1990b) noted that

chinook salmon fry were consistently more sensitive than either the embryos or alevin to selenite.

A total of seven rainbow trout acute studies are presented in Table 1a, but only the two flow-through

measured LC50 values are used to derive the SMAV of 10,488 μg Se/L as recommended by the

Guidelines. The two 96-hr flow-through test LC50 values are 8,800 and 12,500 μg Se/L (Goettl and

Davies 1976; Hodson et al. 1980). As with the coho and chinook salmon, the alevin life stage was less

sensitive to selenite.

Lepomis (bluegill)

The GMAV of 28,500 μg Se/L for the recreationally important bluegill sunfish, Lepomis macrochirus, is

derived from the 96-hr flow-through measured test conducted by Cardwell et al. (1976a,b). The static

measured acute study conduced by Brooke et al. (1985) was not used to calculate the SMAV for this

species, as recommended by the Guidelines.

Se(IV) Freshwater Final Acute Value Determination

Freshwater Species Mean Acute Values (Table 1a) were calculated as geometric means of the available

acute values for selenite, and Genus Mean Acute Values (Table 2a) were then calculated as geometric
15 Draft November 12, 2004 

means of the Species Mean Acute Values. Of the 28 genera for which freshwater mean acute values are

available, the most sensitive genus, Hyalella, is 440 times more sensitive than the most tolerant,

Nephelopsis. The range of sensitivities of the four most sensitive genera spans a factor of 3.7. The

freshwater Final Acute Value (FAV), representing the most sensitive 5th percentile genus, is calculated to

be 514.9 μg/L for selenite using the procedure described in the Guidelines and the Genus Mean Acute

Values in Table 2a. The Final Acute Value is higher than the lowest Species Mean Acute Value (Figure

1).

Acute Toxicity of Se(IV) to Saltwater Animals

Acute toxicity data that can be used to derive a saltwater criterion for selenite are available for 10 species

of invertebrates and eight species of fish that are resident in North America (Table 1b). These 18 species

satisfy the eight family provision specified in the Guidelines. The range of SMAVs for saltwater

invertebrates extends from 255 μg Se/L for juveniles of the bay scallop, Argopecten irradians (Nelson et

al. 1988) to greater than 10,000 μg Se/L for embryos of the blue mussel, Mytilus edulis (Martin et al.

1981) and embryos of the Pacific oyster, Crassostrea gigas (Glickstein 1978; Martin et al. 1981). The

range of SMAVs for fish is slightly wider than that for invertebrates, extending from 599 μg Se/L for

larvae of the haddock, Melanogrammus aeglefinus, to 17,350 μg Se/L for adults of the fourspine

stickleback, Apeltes quadracus (Cardin 1986). No consistent relationship was detected between life

stage of invertebrates or fish and their sensitivity to selenite, and few data are available concerning the

influence of temperature or salinity on the toxicity of selenite to saltwater animals. Acute tests with the

copepod, Acartia tonsa, at 5 and 10°C gave similar results (Lussier 1986). The following text presents a

species-by-species discussion of the eight most sensitive genera, plus all commercially and recreationally

important species. The genera sensitivity ranking is listed in Table 2b.

Argopecten (bay scallop)

The most sensitive saltwater genus is Argopecten, with a GMAV of 255 μg Se/L. The GMAV is derived

from the one available bay scallop (Argopecten irradians) static-renewal unmeasured test conducted by

Nelson et al. (1988) at a salinity of 25 g/kg.
16 Draft November 12, 2004 

Melanogrammus (haddock)

The second most sensitive saltwater genus is Melanogrammus, with a GMAV of 599 μg Se/L. The

GMAV is derived from the one available haddock (Melanogrammus aeglefinus) static unmeasured test

conducted by Cardin (1986) at a salinity of 30 g/kg.

Cancer (dungeness crab)

The third most sensitive saltwater genus is Cancer, with a GMAV of 1,040 μg Se/L. The GMAV is

derived from the one available static unmeasured test conducted by Glickstein (1978) that exposed

Cancer magister to selenium oxide at a salinity of 33.8 g/kg.

Penaeus (brown shrimp)

The fourth most sensitive saltwater genus is Penaeus, with a GMAV of 1,200 μg Se/L. The GMAV is

derived from the one available static unmeasured test conducted by Ward et al. (1981) that exposed 

Penaeus aztecus to sodium selenite at a salinity of 30 g/kg.

Acartia (copepod)

The fifth most sensitive saltwater genus is Acartia, with a GMAV of 1,331 μg Se/L that is derived from

the geometric mean of the A. clausi (2,110 μg Se/L) and A. tonsa (839 μg Se/L) SMAVs. Each of the

SMAVs is derived from one static unmeasured acute test conducted by Lussier (1986) that exposed each

species to selenious acid at a salinity of 30 g/kg.

Americamysis (Mysidopsis) (mysid)

The GMAV of 1,500 μg Se/L for the mysid Americamysis (formerly Mysidopsis) is derived from the one

Americamysis bahia 96-hr flow-through measured test conducted by Ward et al. (1981). The static

unmeasured acute study conduced by U.S. EPA (1978) was not used to calculate the SMAV for this

species as recommended by the Guidelines. The flow-through measured test was conducted with

selenious acid at a salinity of 15-20 g/kg.

Spisula (surf clam)

The seventh most sensitive saltwater genus is Spisula, with a GMAV of 1,900 μg Se/L. The GMAV is

derived from the one available static-renewal unmeasured test conducted by Nelson et al. (1988) that

exposed Spisula solidissima to sodium selenite at a salinity of 25 g/kg.
17 Draft November 12, 2004 

Morone (striped bass)

Five 96-hr static unmeasured tests are available for the striped bass, Morone saxatilis, and the LC50

values ranged from 1,550 to 3,900 μg Se/L (Chapman 1992; Palawski et al. 1985). The geometric

mean of the five values yielded the GMAV of 3,036 μg Se/L. All the tests were conducted with sodium

selenite at a salinity of 1-5 g/kg.

Paralichthys (summer flounder)

The GMAV of 3,497 μg Se/L for the commercially important summer flounder, Paralichthys dentatus, is

derived from one 96-hr static unmeasured acute test conducted by Cardin (1986) that exposed embryos to

selenious acid at a salinity of 30.2 g/kg.

Callinectes (blue crab)

The GMAV of 4,600 μg Se/L for the commercially important blue crab, Callinectes sapidus, is derived

from one static unmeasured acute test conducted by Ward et al. (1981) that exposed juveniles to sodium

selenite at a salinity of 30 g/kg.

Crassostrea (Pacific oyster)

Two static unmeasured tests are available for the commercially important Pacific oyster, Crassostrea

gigas, and the LC50 values were both >10,000 μg Se/L (Glickstein 1978; Martin et al. 1981). The

geometric mean of the two values yielded the GMAV of >10,000 μg Se/L. The tests were conducted

with selenium oxide and sodium selenite at a salinity of 33.8 g/kg.

Mytilus (blue mussel)

The GMAV for the commercially important blue mussel, Mytilus edulis, is also >10,000 μg Se/L, and is

derived from the one static unmeasured acute test conducted by Martin et al. (1981) that exposed

embryos to selenium oxide at a salinity of 33.8 g/kg.

Pseudopleuronectes (winter flounder)

The GMAV of 14,649 μg Se/L for the commercially important winter flounder, Pseudopleuronectes

americanus, is derived from two 96-hr static unmeasured acute tests conducted by Cardin (1986) that

exposed larvae to selenious acid at a salinity of 28-30 g/kg.
18 Draft November 12, 2004 

Se(IV) Saltwater Final Acute Value Determination

Of the 17 genera for which saltwater mean acute values are available for selenite (Table 2b), the most

sensitive genus, Argopectin, is 68 times more sensitive than the most tolerant, Apeltes. The sensitivities

of the four most sensitive genera differ by a factor of 4.7, and these four include three invertebrates and

one fish, of which an invertebrate is the most sensitive of the four. The saltwater Final Acute Value,

representing the most sensitive 5th percentile genus, is 253.4 μg/L for selenite, which is slightly lower

than the lowest Species Mean Acute Value (Figure 2).
19 Draft November 12, 2004 

Acute Toxicity of Selenate

Data that may be used, according to the Guidelines, in the derivation of Final Acute Values for selenate 

are presented in Tables 1a and 1b. The following text presents a brief overview of the acceptable data

obtained for selenate, and includes a discussion of the more sensitive and important species. The genera

sensitivity ranking is listed in Tables 2a and 2b.

Sulfate-dependent Toxicity of Selenate

The toxicity of a number of metals (e.g., copper and cadmium) to aquatic organisms is related to the

concentration of hardness in the water. The toxicity of these metals to many different aquatic species has

been shown to decrease as the hardness concentration increases. A similar relationship also has been

recognized between selenate and dissolved sulfate in freshwater (a similar relationship is not evident

between selenite and sulfate or between either form of selenium and hardness). The studies reviewed in

this document indicate that, as the concentration of sulfate increases, the acute toxicity of selenate is

reduced (less toxic). Selenate acute toxicity tests conducted at different levels of dissolved sulfate are

available with C. dubia, D. magna, H. azteca, G. pseudolimnaeus, chinook salmon and fathead minnows

(Table 1a). These data indicate that, in general, selenate is more toxic to these species in low sulfate

water than in higher sulfate water. 

Sulfate Correction

As discussed in the introduction of this document, sulfate has been shown to compete with selenate in

their uptake into aquatic organisms (Olge and Knight 1996; Riedel and Sanders 1996; Bailey et al. 1995;

Hansen et al. 1993) and affect the acute toxicity of selenate (Brix et al. 2001a). Sulfate is used here as a

correction to the toxicity of selenate. However, it should be emphasized that the sulfate adjustment is not

a precise measure, but an estimation. The variability associated with different life stages, clones and test

conditions of the studies used to determine the sulfate slope all contribute to the uncertainty of the sulfate

correction. In selected cases, insensitive life stages were not used in the analysis (e.g., the eyed-egg and

alevin test results were not used for the chinook salmon). 

Following recommendations in the guidelines (Stephan et al. 1985), an analysis of covariance (Sokal and

Rohlf 1981) was implemented in Microsoft Excel to calculate a common slope for regression lines

projecting the natural logarithm of selenate LC50s as a function of the natural logarithm of sulfate

concentrations. The common regression line is the best estimate of the collective relationship between
20 Draft November 12, 2004 

toxicity and sulfate concentration. With analysis of covariance, different species will be weighted

relative to the number of data points they have. In this case, the fathead minnow has 18 data points out

of the total of 57, the next most frequent species, C. dubia, has 13 data points, and the four remaining

species have eight or fewer data points. 

This analysis of covariance model was fit to the selenate data in Table 1a for the six species for which

definite acute values (“less than” or “greater than” values were not used) were available over a range of

sulfate levels, such that the highest sulfate value was at least three times the lowest, and the highest was

also at least 100 mg/L higher than the lowest (other species in Table 1a either did not meet these criteria

or did not show any sulfate-toxicity trend due to differences in exposure methods, species, age, etc.). A

list of the species and acute toxicity-sulfate values used to estimate the acute sulfate slope is provided in

Appendix A. 

Regression analysis revealed significant, positive slopes for five of six species that had acute values

precisely determined. The slopes for all six species ranged from 0.19 to 0.87, and the common slope for

these six species was 0.5812. An F-test was used to test the null hypothesis that slopes of all species

were equal. This test revealed that the null hypothesis could not be rejected (F5,45 = 2.82, P>0.05). 

Individual slopes were not significantly different than the overall pooled slope (Tukey test, all *q* <3.3,

q 0.05,(2), 47 , 7 = 4.39). Analysis of covariance thus confirmed that it is correct to assume that there is no

significant variation in slopes among species, and that the overall slope is a reasonable estimate of the

relationship between sulfate concentration and selenate toxicity. 

The pooled slope of 0.5812 was used to adjust the freshwater selenate acute values in Table 1a to a

sulfate level of 100 mg/L, except where it was not possible because no sulfate value was reported. 

Species Mean Acute Values (SMAV) were calculated as geometric means of the adjusted acute values

(only the underlined EC50/LC50 species values were used to calculate the respective SMAV). As stated

in the Guidelines (Stephen et al. 1985), flow-through measured study data are normally given preference

over non-flow-through data for a particular species. In certain cases flow-through measured results were

available, yet preference was given to the sensitive life stage for certain species in calculating SMAVs. 

Genus Mean Acute Values (GMAV) at a sulfate level of 100 mg/L were then calculated (Table 1a) as

geometric means of the available freshwater Species Mean Acute Values and ranked (Table 2a). 
21 Draft November 12, 2004 

Acute Toxicity of Se(VI) to Freshwater Animals (Sulfate Adjusted Values)

Acceptable data on the acute effects of selenate in freshwater are available for 12 invertebrate species

and 11 species of fish (Table 1a). These 23 species satisfy the eight family provision of the Guidelines. 

Invertebrates are both the most sensitive and the most tolerant freshwater species to selenate with sulfate

adjusted SMAVs ranging from 593 μg/L for the crustacean, Daphnia pulicaria, to 1,515,616 μg/L for the

leech, Nephelopsis obscura. The selenate SMAVs for fishes range from 10,305 μg/L for the razorback

sucker, Xyrauchen texanus, to 226,320 μg/L for channel catfish, Ictalurus punctatus. The following text

presents a species-by-species discussion of the eight most sensitive genera, plus all commercially and

recreationally important species.

Ceriodaphnia (cladoceran)

The most sensitive freshwater genus is the cladeceran, Ceriodaphnia, with a sulfate adjusted GMAV of

842 μg Se/L. The GMAV is derived from one 48-hr acute flow-through measured test (GLEC 1999). 

Twelve additional tests conducted under non flow-through conditions are also listed in Table 1a (Brix et

al. 2001a,b), but the Guidelines recommend using flow-through measured data in preference to static or

renewal data. 

Hyalella (amphipod)

The second most sensitive freshwater genus is the amphipod, Hyalella, with a sulfate adjusted GMAV of

1,397 μg Se/L. The GMAV is derived from four 96-hr acute flow-through measured tests where the LC50

values ranged from 723 to 4,224 μg Se/L (GLEC 1998). Three tests conducted under non flow-through

conditions are also listed in Table 1a (Adams 1976; Brasher and Ogle 1993; Brix et al. 2001a,b), but are

not used to calculate the SMAV as recommended by the Guidelines.

Daphnia (cladoceran)

The third most sensitive freshwater genus is Daphnia, with a sulfate adjusted GMAV of 1,887 μg Se/L

that is derived from the geometric mean of the D. magna (3,314 μg Se/L), D. pulex (3,420 μg Se/L) and

D. pulicaria (593 μg Se/L) SMAVs. Five static and one static-renewal measured 48-hr studies are

available for D. magna where the LC50 values ranged from 1,955 to 5,093 μg Se/L (Boyum 1984; Brooke

et al. 1985; Dunbar et al. 1983; Ingersol et al. 1990; Maier et al. 1993). 
22 Draft November 12, 2004 

The D. pulex SMAV of 3,420 μg Se/L is based on the 48-hr flow-through measured test conducted by

GLEC (1999) that exposed <24-hr old neonates to sodium selenate. Two static measured tests conducted

by Brix et al. (2001a,b), are not used to calculate the SMAV as recommend by the Guidelines. 

The one available D. pulicaria acute study was conducted by Boyum (1984) that exposed neonates to

sodium selenate for 48 hours under static measured conditions. The resultant 48-hr LC50 value was 593

μg Se/L, which is the most sensitive SMAV for selenate in the database. 

Gammarus (amphipod)

The fourth most sensitive freshwater genus is Gammarus, with a sulfate adjusted GMAV of 2,522 μg

Se/L that is derived from the geometric mean of the G. lacustris (2,747 μg Se/L) and G. pseudolimnaeus

(2,315 μg Se/L) SMAVs. The static measured acute test conduced by Brix et al. (2001a,b) is the only

LC50 value available for G. lacustris.

The G. pseudolimnaeus SMAV of 2,315 μg Se/L is based on five 96-hr flow-through measured tests

conducted by GLEC (1998, 1999). Two static measured acute studies were conduced by Brooke et al.

(1985) and Brooke (1987), but as recommended by the Guidelines, were not used to calculate the SMAV

for this species.

Xyrauchen (razorback sucker)

Six 96-hr static unmeasured tests are available for the razorback sucker, Xyrauchen texanus, and the LC50

values ranged from 7,839 to 16,184 μg Se/L (Buhl and Hamilton 1996; Hamilton 1995; Hamilton and

Buhl 1997a). The geometric mean of the six values yield the GMAV of 10,309 μg Se/L.

Gila (bonytail)

The sixth most sensitive freshwater genus is Gila, with a sulfate adjusted GMAV of 10,560 μg Se/L. 

The GMAV is derived from the one static-unmeasured test conducted with the more sensitive larval stage

(Buhl and Hamilton 1996). Four other static-unmeasured tests were conducted with less sensitive life

stages, but as recommended by the Guidelines, the results were not used to calculate the SMAV for this

species.
23 Draft November 12, 2004 

Pimephales (fathead minnow)

A total of nine fathead minnow acute studies are presented in Table 1a, but only the five flow-through

measured LC50 values are used to derive the sulfate adjusted GMAV of 11,346 μg Se/L. The five flowthrough LC50 values ranged from 7,286 to 18,860 μg Se/L (Spehar 1986; GLEC 1998). The four static

tests are not used to calculate the SMAV as recommended by the Guidelines.

Ptychocheilus (Colorado squawfish)

The eighth most sensitive freshwater genus is Ptychocheilus with a sulfate adjusted GMAV of 18,484 μg

Se/L. The GMAV is derived from the three static-unmeasured test conducted with the sensitive life stage

of Ptychocheilus lucius (Buhl and Hamilton 1996; Hamilton 1995). Three other static-unmeasured tests

were conducted with less sensitive life stages, but as recommended by the Guidelines, the results were

not used to calculate the SMAV for this species.

Oncorhynchus (salmonid)

The sulfate adjusted GMAV of 47,164 μg Se/L for the commercially important salmonid Oncorhynchus

is derived from the geometric mean of the coho salmon (O. kisutch; 29,141 μg Se/L), chinook salmon (O.

tshawytscha; 83,353 μg Se/L) and rainbow trout (O. mykiss; 43,192 μg Se/L) SMAVs. Three static

unmeasured 96-hr studies are used to calculate the coho salmon SMAV where the LC50 values ranged

from 20,963 to 51,935 μg Se/L (Buhl and Hamilton 1991; Hamilton and Buhl 1990b). A fourth coho

salmon LC50 value is available for an acute test initiated with the tolerant alevin life stage (Buhl and

Hamilton 1991), but based on Guideline recommendations this value is not used when data are available

from a more sensitive life stage. 

Five acute chinook salmon static unmeasured 96-hr acute studies conducted with the more sensitive life

stage of the fish are used to determine the sulfate adjusted 83,353 μg Se/L SMAV for the species with

LC50 values ranging from 69,939 to 97,550 μg Se/L (Hamilton and Buhl 1990b). The two acute studies

conducted with the tolerant eyed egg and alevin life stages by the same authors are not used in the

SMAV determination as recommended by the Guidelines. 

A total of four rainbow trout acute studies are presented in Table 1a, but only the results from the two

static tests conducted with the sensitive juvenile life stage were used to calculate the SMAV of 43,192

μg Se/L (Brooke et al. 1985; Buhl and Hamilton 1991). The two test results obtained with less sensitive

life stages were not used as recommended by the Guidelines.
24 Draft November 12, 2004 

Lepomis (bluegill)

The sulfate adjusted GMAV of 216,033 μg Se/L for the recreationally important bluegill sunfish,

Lepomis macrochirus, is derived from the 96-hr static measured test conducted by Brooke et al. (1985)

that exposed juvenile bluegill to sodium selenate.

Ictalurus (channel catfish)

The sulfate adjusted GMAV of 226,320 μg Se/L for the commercially important channel catfish,

Ictalurus punctatus, is derived from the 96-hr static measured test conducted by Brooke et al. (1985) that

exposed juvenile catfish to sodium selenate.

Se(VI) Freshwater Final Acute Value Determination

Of the 18 freshwater genera for which mean sulfate adjusted acute values are available for selenate, the

most sensitive, Ceriodaphnia, is 1,800 times more sensitive than the most tolerant, Nephelopsis. The

range of sensitivities of the four most sensitive genera, all invertebrates, spans a factor of 3.0.

At a sulfate level of 100 mg/L, the freshwater Final Acute Value, representing the most sensitive 5th

percentile genus, was calculated to be 834.4 μg/L for selenate. This Final Acute Value is lower than the

acute value of the most sensitive freshwater species (Table 2a and Figure 3). The resultant freshwater

Criterion Maximum Concentration (CMC) for selenate (in μg/L) = e

(0.5812[ln(sulfate)]+3.357). At a sulfate

level of 100 mg/L this yields 417.2 μg/L, or one-half the FAV. 

Acute Toxicity of Se(VI) to Saltwater Animals

The only species with which acute tests have been conducted on selenate in salt water is the striped bass

(Table 1b). Klauda (1985a, b) obtained 96-hr selenate LC50 values of 9,790 and 85,840 μg/L using flowthrough measured methodology with prolarvae and juvenile striped bass, respectively. In static

unmeasured tests, Chapman (1992) determined selenate 96-hr LC50 values that ranged from 23,700 to

29,000 μg/L using 24 to 32 day posthatch striped bass larvae. The more sensitive prolarvae life stage test

conducted under flow-through conditions is used to yield the SMAV and GMAV of 9,790 μg Se/L for

the striped bass.

Se(VI) Saltwater Final Acute Value Determination

The one saltwater species available for selenate does not satisfy the eight family provision specified in

the Guidelines. Therefore, a saltwater Final Acute Value for selenate cannot be determined. 
25 Draft November 12, 2004 

Comparison of Selenite and Selenate Acute Toxicity

Species Mean Acute Values have been determined for both selenite and selenate with 20 freshwater

species (Table 3a) and one saltwater species (Table 3b). Of these 21 species, 20 are more sensitive to

Se(IV). Only the amphipod, Gammarus pseudolimnaeus, is more sensitive to Se (VI), and is in the

sensitive portion of the Table 3a distribution. Consistent with the acute toxicity sensitivity pattern, the

FAV for Se(VI) is higher than the FAV for Se (IV).
26 Draft November 12, 2004 

Table 1a. Acute Toxicity o f Selenium to Freshwater Animals

Species Methoda Chemical

Hardness

(mg/L as

 CaCO3

) 

LC50

or EC50

 (μg/L)b

Species Mean

Acute Value

 (μg/L) Reference

FRESHWATER SPECIES

Selenite

Hydra (adult),

Hydra sp.

S, M Sodium

selenite

- 1,700 1,700 Brooke et al. 1985

Worm,

Tubifex tubifex

R, U Sodium

selenite

245 7,710 7,710 Khangarot 1991

Leech (adult),

Nephelopsis obscura

S, M Sodium

selenite

49.8 203,000 203,000 Brooke et al. 1985

Snail (adult),

Aplexa hypnorum

S, M Sodium

selenite

50.6 53,000 - Brooke et al. 1985

Snail (adult),

Aplexa hypnorum

S, M Sodium

selenite

49.8 23,000 34,914 Brooke et al. 1985

Snail,

Physa sp.

S, U Sodium

selenite

45.7 24,100 24,100 Reading 1979

Cladoceran (<24 hr),

Ceriodaphnia dubia

F, M Sodium

selenite

127

(sulfate=25)

440 440 GLEC 1999

Cladoceran (<24 hr),

Ceriodaphnia affinis

S, U Sodium

selenite

100.8 600 - Owsley 1984;

Owsley and

McCauley 1986

Cladoceran (36-60 hr),

Ceriodaphnia affinis

S, U Sodium

selenite

100.8 720 - Owsley 1984

Cladoceran (84-108 hr),

Ceriodaphnia affinis

S, U Sodium

selenite

100.8 640 - Owsley 1984

Cladoceran (72-120 hr),

Ceriodaphnia affinis

S, U Sodium

selenite

100.8 <480 <603.6 Owsley 1984

Cladoceran,

Daphnia magna

S, U Sodium

selenite

214 2,500 - Bringmann and

Kuhn 1959a

Cladoceran,

Daphnia magna

S, U Selenious 

acidc

72 430 - LeBlanc 1980

Cladoceran,

Daphnia magna

S, M Sodium

selenite

129.5 1,100 - Dunbar et al. 1983

Cladoceran,

Daphnia magna

S, M Sodium

selenite

138 450 - Boyum 1984

Cladoceran (<24 hr),

Daphnia magna

S, U Sodium

selenite

- 215 - Adams and

Heidolph 1985

Cladoceran (<24 hr),

Daphnia magna

S, U Sodium

selenite

40 870 - Mayer and

Ellersieck 1986

Cladoceran (<24 hr),

Daphnia magna

S, U Sodium

selenite

280 2,370 - Mayer and

Ellersieck 1986
Table 1a. Acute Toxicity of Selenium to Freshwater Animals (continued)

Species Methoda Chemical

Hardness

(mg/L as

 CaCO3) 

LC50

or EC50

 (μg/L)b

Species Mean

Acute Value

 (μg/L) Reference

27 Draft November 12, 2004 

Cladoceran,

Daphnia magna

S, M Sodium

selenite

45.5 700 - Ingersoll et al.

1990

Cladoceran,

Daphnia magna

S, M Sodium

selenite

136 3,020 - Ingersoll et al.

1990

Cladoceran (<24 hr),

Daphnia magna

R, M Sodium

selenite

80-100 550 - Maier et al. 1993

Cladoceran,

Daphnia magna

S, M Selenious

 acid

220d

1,220 9,05.3 Kimball,

Manuscript

Cladoceran,

Daphnia pulex

S, M Sodium

selenite

46.4 3,870 - Reading 1979;

Reading and

Buikema 1983

Cladoceran (<24 hr),

Daphnia pulex

F, M Sodium

selenite

128

(sulfate=25)

1,987 1,987 GLEC 1999

Amphipod (adult),

Gammarus

pseudolimnaeus

S, M Sodium

selenite

48.3 4,300 - Brooke et al. 1985

Amphipod (adult),

Gammarus

pseudolimnaeus

S, M Sodium

selenite

53.6 1,700 - Brooke 1987

Amphipod,

Gammarus

pseudolimnaeus

F, M Sodium

selenite

139

(sulfate=24)

2,260 -- GLEC 1998

Amphipod,

Gammarus

pseudolimnaeus

F, M Sodium

selenite

137

(sulfate=138)

3,130 -- GLEC 1998

Amphipod,

Gammarus

pseudolimnaeus

F, M Sodium

selenite

144

(sulfate=326)

1,800 -- GLEC 1998

Amphipod,

Gammarus

pseudolimnaeus

F, M Sodium

selenite

138

(sulfate=758)

3,710 -- GLEC 1998

Amphipod (adult),

Gammarus

pseudolimnaeus

F, M Sodium

selenite

128

(sulfate=25)

10,950 3,489 GLEC 1999

Amphipod 

(2 mm length),

Hyalella azteca

R, M Sodium

selenite

133 420 - Brasher and Ogle

1993

Amphipod,

Hyalella azteca

F, M Sodium

selenite

329 340 - Halter et al. 1980

Amphipod,

Hyalella azteca

F, M Sodium

selenite

132

(sulfate=64)

670 -- GLEC 1998
Table 1a. Acute Toxicity of Selenium to Freshwater Animals (continued)

Species Methoda Chemical

Hardness

(mg/L as

 CaCO3) 

LC50

or EC50

 (μg/L)b

Species Mean

Acute Value

 (μg/L) Reference

28 Draft November 12, 2004 

Amphipod,

Hyalella azteca

F, M Sodium

selenite

132

(sulfate=138)

<350 -- GLEC 1998

Amphipod,

Hyalella azteca

F, M Sodium

selenite

138

(sulfate=359)

<460 -- GLEC 1998

Amphipod,

Hyalella azteca

F, M Sodium

selenite

138

(sulfate=642)

570 461.4 GLEC 1998

Midge (4th instar),

Chironomus decorus

R, M Sodium

selenite

85 48,200 48,200 Maier and Knight

1993

Midge,

Chironomus plumosus

S, U Sodium

selenite

39 24,150 - Mayer and

Ellersieck 1986

Midge,

Chironomus plumosus

S, U Sodium

selenite

280 27,850 25,934 Mayer and

Ellersieck 1986

Midge,

Tanytarsus dissimilis

F, M Selenium

dioxide

48 42,500 42,500 Call et al. 1983

Coho salmon (0.5 g),

Oncorhynchus kisutch

S, U Sodium

selenite

211 7,800 - Hamilton and

Buhl 1990b

Coho salmon (2.6 g),

Oncorhynchus kisutch

S, U Sodium

selenite

333 13,600 - Hamilton and

Buhl 1990b

Coho salmon (alevin),

Oncorhynchus kisutch

S, U Sodium

selenite

41 35,560f

- Buhl and

Hamilton 1991

Coho salmon (juvenile),

Oncorhynchus kisutch

S, U Sodium

selenite

41 3,578 7,240 Buhl and

Hamilton 1991

Chinook salmon (0.7 g),

Oncorhynchus

tshawytscha

S, U Sodium

selenite

211 14,800 - Hamilton and

Buhl 1990b

Chinook salmon (0.5 g),

Oncorhynchus

tshawytscha

S, U Sodium

selenite

211 13,000 - Hamilton and

Buhl 1990b

Chinook salmon (1.6 g),

Oncorhynchus

tshawytscha

S, U Sodium

selenite

333 23,100 - Hamilton and

Buhl 1990b

Chinook salmon (1.6 g),

Oncorhynchus

tshawytscha

S, U Sodium

selenite

333 23,400 - Hamilton and

Buhl 1990b

Chinook salmon 

(eyed egg),

Oncorhynchus

tshawytscha

S, U Sodium

selenite

41.7 >348,320f- Hamilton and

Buhl 1990b

Chinook salmon (alevin),

Oncorhynchus

tshawytscha

S, U Sodium

selenite

41.7 64,690f- Hamilton and

Buhl 1990b
Table 1a. Acute Toxicity of Selenium to Freshwater Animals (continued)

Species Methoda Chemical

Hardness

(mg/L as

 CaCO3) 

LC50

or EC50

 (μg/L)b

Species Mean

Acute Value

 (μg/L) Reference

29 Draft November 12, 2004 

Chinook salmon (0.31 g),

Oncorhynchus

tshawytscha

S, U Sodium

selenite

41.7 16,980 - Hamilton and

Buhl 1990b

Chinook salmon (0.46 g),

Oncorhynchus

tshawytscha

S, U Sodium

selenite

41.7 8,150 15,596 Hamilton and

Buhl 1990b

Rainbow trout,

Oncorhynchus mykiss

S, U Sodium

selenite

330 4,500 - Adams 1976

Rainbow trout,

Oncorhynchus mykiss

S, U Sodium

selenite

330 4,200 - Adams 1976

Rainbow trout,

Oncorhynchus mykiss

S, U Sodium

selenite

272 1,800 - Hunn et al. 1987

Rainbow trout 

(alevin),

Oncorhynchus mykiss

S, U Sodium

selenite

41 118,000 - Buhl and

Hamilton 1991

Rainbow trout 

(juvenile),

Oncorhynchus mykiss

S, U Sodium

selenite

41 9,000 - Buhl and

Hamilton 1991

Rainbow trout,

Oncorhynchus mykiss

F, M Sodium

selenite

30 12,500 - Goettl and Davies

1976

Rainbow trout,

Oncorhynchus mykiss

F, M Sodium

selenite

135 8,800 10,488 Hodson et al. 1980

Brook trout 

(adult),

Salvelinus fontinalis

F, M Selenium

dioxide

157 10,200 10,200 Cardwell et al.

1976a,b

Arctic grayling 

(alevin),

Thymallus arcticus

S, U Sodium

selenite

41 34,732f

- Buhl and

Hamilton 1991

Arctic grayling 

(juvenile),

Thymallus arcticus

S, U Sodium

selenite

41 15,675 15,675 Buhl and

Hamilton 1991

Goldfish,

Carassius auratus

F, M Selenium

dioxide

157 26,100 26,100 Cardwell et al.

1976a,b

Common carp,

Cyprinus carpio

R, U - - 35,000 35,000 Sato et al. 1980

Golden shiner,

Notemigonus crysoleucas

F, M Sodium

selenite

72.2 11,200 11,200 Hartwell et al.

1989

Fathead minnow,

Pimephales promelas

S, U Sodium

selenite

312

 (13°C)

10,500 - Adams 1976

Fathead minnow,

Pimephales promelas

S, U Sodium

selenite

312

(13°C)

11,300 - Adams 1976
Table 1a. Acute Toxicity of Selenium to Freshwater Animals (continued)

Species Methoda Chemical

Hardness

(mg/L as

 CaCO3) 

LC50

or EC50

 (μg/L)b

Species Mean

Acute Value

 (μg/L) Reference

30 Draft November 12, 2004 

Fathead minnow,

Pimephales promelas

S, U Sodium

selenite

303

(20°C)

6,000 - Adams 1976

Fathead minnow,

Pimephales promelas

S, U Sodium

selenite

303

(20°C)

7,400 - Adams 1976

Fathead minnow,

Pimephales promelas

S, U Sodium

selenite

292

(25°C)

3,400 - Adams 1976

Fathead minnow,

Pimephales promelas

S, U Sodium

selenite

292

(25°C)

2,200 - Adams 1976

Fathead minnow 

(30 days),

Pimephales promelas

S, M Sodium

selenite

51.1 1,700 - Brooke et al. 1985

Fathead minnow

(juvenile),

Pimephales promelas

S, U Sodium

selenite

40 7,760 - Mayer and

Ellersieck 1986

Fathead minnow 

(fry),

Pimephales promelas

F, M Selenium

dioxide

157 2,100 - Cardwell et al.

1976a,b

Fathead minnow

(juvenile),

Pimephales promelas

F, M Selenium

dioxide

157 5,200 - Cardwell et al.

1976a,b

Fathead minnow,

Pimephales promelas

F, M Sodium

selenite

131

(sulfate=24)

3,670 -- GLEC 1998

Fathead minnow,

Pimephales promelas

F, M Sodium

selenite

131

(sulfate=160)

2,920 -- GLEC 1998

Fathead minnow,

Pimephales promelas

F, M Sodium

selenite

145

(sulfate=214)

3,390 -- GLEC 1998

Fathead minnow,

Pimephales promelas

F, M Sodium

selenite

140

(sulfate=870)

2,380 - GLEC 1998

Fathead minnow,

Pimephales promelas

F, M Selenious

 acid

220d

620 - Kimball,

Manuscript

Fathead minnow,

Pimephales promelas

F, M Selenious

acid

220d

970 2,209 Kimball,

Manuscript

Colorado squawfish 

(fry),

Ptychocheilus lucius

S, U Sodium

selenite

197 6,398 - Hamilton 1995

Colorado squawfish 

(0.4-1.1 g juvenile),

Ptychocheilus lucius

S, U Sodium

selenite

197 16,452 - Hamilton 1995

Colorado squawfish 

(1.7 g juvenile),

Ptychocheilus lucius

S, U Sodium

selenite

197 14,624 - Hamilton 1995
Table 1a. Acute Toxicity of Selenium to Freshwater Animals (continued)

Species Methoda Chemical

Hardness

(mg/L as

 CaCO3) 

LC50

or EC50

 (μg/L)b

Species Mean

Acute Value

 (μg/L) Reference

31 Draft November 12, 2004 

Colorado squawfish

(larva),

Ptychocheilus lucius

S, U Sodium

selenite

199 7,960 - Buhl and

Hamilton 1996

Colorado squawfish

(juvenile),

Ptychocheilus lucius

S, U Sodium

selenite

199 17,350 - Buhl and

Hamilton 1996

Colorado squawfish 

(0.024-0.047 g),

Ptychocheilus lucius

S, U Sodium

selenite

144 20,700 12,801 Hamilton and

Buhl 1997a

Bonytail (fry),

Gila elegans

S, U Sodium

selenite

197 8,680 - Hamilton 1995

Bonytail (1.1 g juvenile),

Gila elegans

S, U Sodium

selenite

197 7,769 - Hamilton 1995

Bonytail (2.6 g juvenile),

Gila elegans

S, U Sodium

selenite

197 6,855 - Hamilton 1995

Bonytail (larva),

Gila elegans

S, U Sodium

selenite

199 14,490 - Buhl and

Hamilton 1996

Bonytail (juvenile),

Gila elegans

S, U Sodium

selenite

199 12,870 9,708 Buhl and

Hamilton 1996

Razorback sucker 

(fry),

Xyrauchen texanus

S, U Sodium

selenite

197 6,855 - Hamilton 1995

Razorback sucker 

(0.9 g juvenile),

Xyrauchen texanus

S, U Sodium

selenite

197 4,067 - Hamilton 1995

Razorback sucker 

(2.0 g juvenile),

Xyrauchen texanus

S, U Sodium

selenite

197 7,312 - Hamilton 1995

Razorback sucker 

(larva),

Xyrauchen texanus

S, U Sodium

selenite

199 10,450 - Buhl and

Hamilton 1996

Razorback sucker

(juvenile),

Xyrauchen texanus

S, U Sodium

selenite

199 8,520 - Buhl and

Hamilton 1996

Razorback sucker

(0.006-0.042 g),

Xyrauchen texanus

S, U Sodium

selenite

144 11,300 7,679 Hamilton and

Buhl 1997a

White sucker,

Catostomus commersoni

F, M Sodium

selenite

10.2 29,000 - Klaverkamp et al.

1983a

White sucker,

Catostomus commersoni

F, M Sodium

selenite

18 31,400 30,176 Duncan and

Klaverkamp 1983
Table 1a. Acute Toxicity of Selenium to Freshwater Animals (continued)

Species Methoda Chemical

Hardness

(mg/L as

 CaCO3) 

LC50

or EC50

 (μg/L)b

Species Mean

Acute Value

 (μg/L) Reference

32 Draft November 12, 2004 

Flannelmouth sucker 

(12-13 days),

Catostomus latipinnis

S, U Sodium

selenite

144 19,100 19,100 Hamilton and

Buhl 1997b

Striped bass (63 days),

Morone saxatilis

S, U Sodium

selenite

40 1,325 - Palawski et al.

1985

Striped bass (63 days),

Morone saxatilis

S, U Sodium

selenite

285 2,400 1,783 Palawski et al.

1985

Channel catfish (juvenile),

Ictalurus punctatus

S, M Sodium

selenite

49.8 16,000 - Brooke et al. 1985

Channel catfish (juvenile),

Ictalurus punctatus

S, U Sodium

selenite

41 4,110 - Mayer and

Ellersieck 1986

Channel catfish,

Ictalurus punctatus

F, M Selenium

dioxide

157 13,600 13,600 Cardwell et al.

1976a,b

Flagfish,

Jordanella floridae

F, M Selenium

dioxide

157 6,500 6,500 Cardwell et al.

1976a,b

Mosquitofish,

Gambusia affinis

S, U Sodium

selenite

45.7 12,600 12,600 Reading 1979

Bluegill (juvenile),

Lepomis macrochirus

S, M Sodium

selenite

50.5 12,000 - Brooke et al. 1985

Bluegill, 

Lepomis macrochirus

F, M Selenium

dioxide

157 28,500 28,500 Cardwell et al.

1976a,b

Yellow perch, 

Perca flavescens

F, M Sodium

selenite

10.2 11,700 11,700 Klaverkamp et al.

1983a

Species Methoda Chemical

Hardness

(mg/L as

 CaCO3) 

LC50

or EC50

 (μg/L)b

LC50 or EC50

Adj. To 

Sulfate = 100

 (μg/L)

Species Mean

Acute Value at

Sulfate = 100

 (μg/L) Reference

FRESHWATER SPECIES

Selenate

Hydra (adult),

Hydra sp.

S, M Sodium

selenate

53.6

(sulfate=12)

7300 25,032 25,032 Brooke et al.

1985

Leech (adult), 

Nephelopsis

obscura

S, M Sodium

selenate

49.3

(sulfate=12)

442000 1,515,661 1,515,661 Brooke et al.

1985

Snail, 

Aplexa hypnorum

S, M Sodium

selenate

51.0

(sulfate=12)

193000 661,816 661,816 Brooke et al.

1985
Table 1a. Acute Toxicity of Selenium to Freshwater Animals (continued)

Species Methoda Chemical

Hardness

(mg/L as

 CaCO3) 

LC50

or EC50

 (μg/L)b

LC50 or EC50

Adj. To 

Sulfate = 100

 (μg/L)

Species Mean

Acute Value at

Sulfate = 100

 (μg/L) Reference

33 Draft November 12, 2004 

Cladoceran 

(<24 hr),

Ceriodaphnia

dubia

S, M Sodium

selenate

52

(sulfate=52)

1967 2,877 -- Brix et al.

2001a,b

Cladoceran 

(<24 hr),

Ceriodaphnia

dubia

S, M Sodium

selenate

52

(sulfate=55)

1864 2,638 -- Brix et al.

2001a,b

Cladoceran 

(<24 hr),

Ceriodaphnia

dubia

S, M Sodium

selenate

52

(sulfate=31)

1078 2,129 -- Brix et al.

2001a,b

Cladoceran

 (<24 hr),

Ceriodaphnia

dubia

S, M Sodium

selenate

52

(sulfate=38)

580 1,018 -- Brix et al.

2001a,b

Cladoceran 

(<24 hr),

Ceriodaphnia

dubia

S, M Sodium

selenate

52

(sulfate=98)

1822 1,844 -- Brix et al.

2001a,b

Cladoceran

 (<24 hr),

Ceriodaphnia

dubia

S, M Sodium

selenate

52

(sulfate=98)

1728 1,748 -- Brix et al.

2001a,b

Cladoceran

 (<24 hr),

Ceriodaphnia

dubia

S, M Sodium

selenate

52

(sulfate=213)

1453 936 -- Brix et al.

2001a,b

Cladoceran

 (<24 hr),

Ceriodaphnia

dubia

S, M Sodium

selenate

52

(sulfate=217)

2812 1,793 -- Brix et al.

2001a,b

Cladoceran

 (<24 hr),

Ceriodaphnia

dubia

S, M Sodium

selenate

52

(sulfate=378)

5553 2,564 -- Brix et al.

2001a,b

Cladoceran 

(<24 hr),

Ceriodaphnia

dubia

S, M Sodium

selenate

52

(sulfate=378)

5481 2,531 -- Brix et al.

2001a,b

Cladoceran 

(<24 hr),

Ceriodaphnia

dubia

S, M Sodium

selenate

52

(sulfate=926)

9157 2,512 -- Brix et al.

2001a,b
Table 1a. Acute Toxicity of Selenium to Freshwater Animals (continued)

Species Methoda Chemical

Hardness

(mg/L as

 CaCO3) 

LC50

or EC50

 (μg/L)b

LC50 or EC50

Adj. To 

Sulfate = 100

 (μg/L)

Species Mean

Acute Value at

Sulfate = 100

 (μg/L) Reference

34 Draft November 12, 2004 

Cladoceran

(<24 hr),

Ceriodaphnia

dubia

S, M Sodium

selenate

52

(sulfate=1205)

9311 2,191 -- Brix et al.

2001a,b

Cladoceran 

(<24 hr),

Ceriodaphnia

dubia

F, M Sodium

selenate

127

(sulfate=25)

376 842 842 GLEC 1999

Cladoceran, 

Daphnia magna

S, M Sodium

selenate

129.5

(sulfate=163)

5300 3,990 -- Dunbar et al.

1983

Cladoceran, 

Daphnia magna

S, M Sodium

selenate

138

(sulfate=22)

1010 2,435 -- Boyum 1984

Cladoceran, 

Daphnia magna

S, M Sodium

selenate

48.1

(sulfate=12)

570 1,955 -- Brooke et al.

1985

Cladoceran,

Daphnia magna

S, M Sodium

selenate

45.5

(sulfate=41)

2560 4,298 -- Ingersoll et al.

1990

Cladoceran,

Daphnia magna

S, M Sodium

selenate

136

(sulfate=68)

4070 5,093 -- Ingersoll et al.

1990

Cladoceran (<24

hr),

Daphnia magna

R, M Sodium

selenate

80-100

(sulfate=82)

2840 3,187 3,314 Maier et al. 1993

Cladoceran (<24

hr),

Daphnia pulex

S, M Sodium

selenate

52

(sulfate=54)

10123 14,482 -- Brix et al.

2001a,b

Cladoceran (<24

hr),

Daphnia pulex

S, M Sodium

selenate

52

(sulfate=38)

8126 14,233 -- Brix et al.

2001a,b

Cladoceran (<24

hr),

Daphnia pulex

F, M Sodium

selenate

147

(sulfate=25)

1528 3,420 3,420 GLEC 1999

Cladoceran, 

Daphnia pulicaria

S, M Sodium

selenate

138

(sulfate=22)

246 593 593 Boyum 1984

Amphipod (8-12

mm), 

Gammarus

lacustris

S, M Sodium

selenate

116

(sulfate=120)

3054 2,747 2,747 Brix et al.

2001a,b

Amphipod (adult), 

Gammarus

pseudolimnaeus

S, M Sodium

selenate

46.1

(sulfate=12)

75 257 -- Brooke et al.

1985

Amphipod (adult),

Gammarus

pseudolimnaeus

S, M Sodium

selenate

51.0

(sulfate=12)

57 196 -- Brooke 1987
Table 1a. Acute Toxicity of Selenium to Freshwater Animals (continued)

Species Methoda Chemical

Hardness

(mg/L as

 CaCO3) 

LC50

or EC50

 (μg/L)b

LC50 or EC50

Adj. To 

Sulfate = 100

 (μg/L)

Species Mean

Acute Value at

Sulfate = 100

 (μg/L) Reference

35 Draft November 12, 2004 

Amphipod,

Gammarus

pseudolimnaeus

F, M Sodium

selenate

139

(sulfate=25)

1180 2,641 -- GLEC 1998

Amphipod,

Gammarus

pseudolimnaeus

F, M Sodium

selenate

132

(sulfate=125)

2870 2,521 -- GLEC 1998

Amphipod,

Gammarus

pseudolimnaeus

F, M Sodium

selenate

137

(sulfate=367)

3710 1,743 -- GLEC 1998

Amphipod,

Gammarus

pseudolimnaeus

F, M Sodium

selenate

134

(sulfate=635)

3270 1,167 -- GLEC 1998

Amphipod (adult),

Gammarus

pseudolimnaeus

F, M Sodium

selenate

131

(sulfate=25)

2191 4,904 2,315 GLEC 1999

Amphipod,

Hyalella azteca

F, U Sodium

selenate

336.8

(sulfate NA)

760 -- -- Adams 1976

Amphipod 

(2 mm length),

Hyalella azteca

R, M Sodium

selenate

133

(sulfate=13)

1031 3,375 - Brasher and Ogle

1993

Amphipod 

(7-10 days),

Hyalella azteca

S, M Sodium

selenate

52

(sulfate=55)

1424 2,021 -- Brix et al.

2001a,b

Amphipod,

Hyalella azteca

F, M Sodium

selenate

143

(sulfate=40)

2480 4,224 -- GLEC 1998

Amphipod,

Hyalella azteca

F, M Sodium

selenate

132

(sulfate=125)

1350 1,186 -- GLEC 1998

Amphipod,

Hyalella azteca

F, M Sodium

selenate

137

(sulfate=367)

1540 723 -- GLEC 1998

Amphipod,

Hyalella azteca

F, M Sodium

selenate

133

(sulfate=822)

3580 1,052 1,397 GLEC 1998

Midge (4th instar),

Chironomus

decorus

R, M Sodium

selenate

85

(sulfate=27)

23700 50,727 50,727 Maier and Knight

1993

Midge (3rd instar),

Paratanytarsus

parthenogeneticus

S, M Sodium

selenate

49.4

(sulfate=12)

20000 68,582 68,582 Brooke et al.

1985

Coho salmon 

(0.5 g),

Oncorhynchus

kisutch

S, U Sodium

selenate

211

(sulfate=185)

32500 22,730 -- Hamilton and

Buhl 1990b
Table 1a. Acute Toxicity of Selenium to Freshwater Animals (continued)

Species Methoda Chemical

Hardness

(mg/L as

 CaCO3) 

LC50

or EC50

 (μg/L)b

LC50 or EC50

Adj. To 

Sulfate = 100

 (μg/L)

Species Mean

Acute Value at

Sulfate = 100

 (μg/L) Reference

36 Draft November 12, 2004 

Coho salmon 

(1.7 g),

Oncorhynchus

kisutch

S, U Sodium

selenate

333

(sulfate=291)

39000 20,963 -- Hamilton and

Buhl 1990b

Coho salmon 

(alevin),

Oncorhynchus

kisutch

S, U Sodium

selenate

41

(sulfate=41)

158,422f265,990f-- Buhl and

Hamilton 1991

Coho salmon 

(juvenile),

Oncorhynchus

kisutch

S, U Sodium

selenate

41

(sulfate=41)

30932 51,935 29,141 Buhl and

Hamilton 1991

Chinook salmon 

(0.7 g),

Oncorhynchus

tshawytscha

S, U Sodium

selenate

211

(sulfate=185)

121000 84,626 -- Hamilton and

Buhl 1990b

Chinook salmon 

(0.5 g),

Oncorhynchus

tshawytscha

S, U Sodium

selenate

211

(sulfate=185)

100000 69,939 -- Hamilton and

Buhl 1990b

Chinook salmon 

(1.6 g),

Oncorhynchus

tshawytscha

S, U Sodium

selenate

333

(sulfate=291)

180000 96,752 -- Hamilton and

Buhl 1990b

Chinook salmon 

(1.6 g),

Oncorhynchus

tshawytscha

S, U Sodium

selenate

333

(sulfate=291)

134000 72,026 -- Hamilton and

Buhl 1990b

Chinook salmon

(eyed egg),

Oncorhynchus

tshawytscha

S, U Sodium

selenate

41.7

(sulfate=47)

>552,000f >856,083f-- Hamilton and

Buhl 1990b

Chinook salmon 

(alevin),

Oncorhynchus

tshawytscha

S, U Sodium

selenate

41.7

(sulfate=47)

>176,640f >273,947f-- Hamilton and

Buhl 1990b

Chinook salmon 

(0.31 g),

Oncorhynchus

tshawytscha

S, U Sodium

selenate

41.7

(sulfate=47)

62900 97,550 83,353 Hamilton and

Buhl 1990b

Rainbow trout 

(juvenile),

Oncorhynchus

mykiss

S, M Sodium

selenate

51.0

(sulfate=12)

24000 82,298 -- Brooke et al.

1985
Table 1a. Acute Toxicity of Selenium to Freshwater Animals (continued)

Species Methoda Chemical

Hardness

(mg/L as

 CaCO3) 

LC50

or EC50

 (μg/L)b

LC50 or EC50

Adj. To 

Sulfate = 100

 (μg/L)

Species Mean

Acute Value at

Sulfate = 100

 (μg/L) Reference

37 Draft November 12, 2004 

Rainbow trout 

(alevin),

Oncorhynchus

mykiss

S, U Sodium

selenate

41

(sulfate=41)

196460 329,856f

-- Buhl and

Hamilton 1991

Rainbow trout, 

Oncorhynchus

mykiss

F, M Sodium

selenate

45

(sulfate=12)

47000 161,168f-- Spehar 1986

Rainbow trout 

(juvenile),

Oncorhynchus

mykiss

S, U Sodium

selenate

41

(sulfate=41)

13501 22,668 43,192 Buhl and

Hamilton 1991

Arctic grayling 

(alevin),

Thymallus arcticus

S, U Sodium

selenate

41

(sulfate=41)

41800 70,182 -- Buhl and

Hamilton 1991

Arctic grayling 

(juvenile),

Thymallus arcticus

S, U Sodium

selenate

41

(sulfate=41)

75240 126,328 94,159 Buhl and

Hamilton 1991

Fathead minnow,

Pimephales

promeles

S, U Sodium

selenate

323

(sulfate NA)

11800 -- -- Adams 1976

Fathead minnow,

Pimephales

promeles

S, U Sodium

selenate

323

(sulfate NA)

11000 -- -- Adams 1976

Fathead minnow,

Pimephales

promeles

S, U Sodium

selenate

323

(sulfate NA)

12500 -- -- Adams 1976

Fathead minnow

(juvenile),

Pimephales

promelas

S, M Sodium

selenate

47.9

(sulfate =12)

2300 7,887 -- Brooke et al.

1985

Fathead minnow,

Pimephales

promelas

F, M Sodium

selenate

46

(sulfate =12)

5500 18,860 -- Spehar 1986

Fathead minnow,

Pimephales

promelas

F, M Sodium

selenate

136

(sulfate=24)

6210 14,236 -- GLEC 1998

Fathead minnow,

Pimephales

promelas

F, M Sodium

selenate

127

(sulfate=160)

10800 8,218 -- GLEC 1998

Fathead minnow,

Pimephales

promelas

F, M Sodium

selenate

131

(sulfate=474)

18000 7,286 -- GLEC 1998
Table 1a. Acute Toxicity of Selenium to Freshwater Animals (continued)

Species Methoda Chemical

Hardness

(mg/L as

 CaCO3) 

LC50

or EC50

 (μg/L)b

LC50 or EC50

Adj. To 

Sulfate = 100

 (μg/L)

Species Mean

Acute Value at

Sulfate = 100

 (μg/L) Reference

38 Draft November 12, 2004 

Fathead minnow,

Pimephales

promelas

F, M Sodium

selenate

147

(sulfate=906)

42100 11,695 11,346 GLEC 1998

Colorado

squawfish 

(fry),

Ptychocheilus

lucius

S, U Sodium

selenate

196

(sulfate=164)

27588 20,694 -- Hamilton 1995

Colorado

squawfish

(0.4-1.1 g

juvenile),

Ptychocheilus

lucius

S, U Sodium

selenate

196

(sulfate=164)

119548 89,676f

-- Hamilton 1995

Colorado

squawfish 

(1.7 g juvenile),

Ptychocheilus

lucius

S, U Sodium

selenate

196

(sulfate=164)

138358 103,786f

-- Hamilton 1995

Colorado

squawfish (larva),

Ptychocheilus

lucius

S, U Sodium

selenate

199

(sulfate=174)

13580 9,842 -- Buhl and

Hamilton 1996

Colorado

squawfish

(0.024-0.047 g),

Ptychocheilus

lucius

S, U Sodium

selenate

144

(sulfate=97)

88000 89,572f-- Hamilton and

Buhl 1997a

Colorado

squawfish

(juvenile),

Ptychocheilus

lucius

S, U Sodium

selenate

199

(sulfate=174)

42780 31,005 18,484 Buhl and

Hamilton 1996

Bonytail 

(fry),

Gila elegans

S, U Sodium

selenate

196

(sulfate=164)

22990 17,245f

-- Hamilton 1995

Bonytail 

(1.1 g juvenile),

Gila elegans

S, U Sodium

selenate

196

(sulfate=164)

102828 77,134f

-- Hamilton 1995

Bonytail 

(2.6 g juvenile),

Gila elegans

S, U Sodium

selenate

196

(sulfate=164)

90706 68,041f-- Hamilton 1995

Bonytail 

(juvenile),

Gila elegans

S, U Sodium

selenate

199

(sulfate=174)

24010 17,401f

-- Buhl and

Hamilton 1996
Table 1a. Acute Toxicity of Selenium to Freshwater Animals (continued)

Species Methoda Chemical

Hardness

(mg/L as

 CaCO3) 

LC50

or EC50

 (μg/L)b

LC50 or EC50

Adj. To 

Sulfate = 100

 (μg/L)

Species Mean

Acute Value at

Sulfate = 100

 (μg/L) Reference

39 Draft November 12, 2004 

Bonytail 

(larva),

Gila elegans

S, U Sodium

selenate

199

(sulfate=174)

14570 10,560 10,560 Buhl and

Hamilton 1996

Razorback sucker 

(fry),

Xyrauchen texanus

S, U Sodium

selenate

196

(sulfate=164)

20064 15,051 -- Hamilton 1995

Razorback sucker 

(0.9 g juvenile),

Xyrauchen texanus

S, U Sodium

selenate

196

(sulfate=164)

15048 11,288 - - Hamilton 1995

Razorback sucker 

(2.0 g juvenile),

Xyrauchen texanus

S, U Sodium

selenate

196

(sulfate=164)

10450 7,839 -- Hamilton 1995

Razorback sucker 

(larva),

Xyrauchen texanus

S, U Sodium

selenate

199

(sulfate=174)

13910 10,081 -- Buhl and

Hamilton 1996

Razorback sucker

(juvenile),

Xyrauchen texanus

S, U Sodium

selenate

199

(sulfate=174)

7620 5,523 -- Buhl and

Hamilton 1996

Razorback sucker 

(0.006-0.042 g),

Xyrauchen texanus

S, U Sodium

selenate

144

(sulfate=97)

15900 16,184 10,309 Hamilton and

Buhl 1997a

Flannelmouth

sucker

(12-13 days),

Catostomus

latipinnis

S, U Sodium

selenate

144

(sulfate=97)

26900 27,380 27,380 Hamilton and

Buhl 1997b

Channel catfish

(juvenile),

Ictalurus

punctatus

S, M Sodium

selenate

51.0

(sulfate=12)

66000 226,320 226,320 Brooke et al.

1985

Bluegill (juvenile),

Lepomis

macrochirus

S, M Sodium

selenate

50.4

(sulfate=12)

63000 216,033 216,033 Brooke et al.

1985

a

 S = static; R = renewal; F = flow-through; M = measured; U = unmeasured.

b

 Concentration of selenium, not the chemical. Note: The values underlined in this column were used to calculate the SMAV

 for the respective species.

c

 Reported by Barrows et al. (1980) in work performed in the same laboratory under the same contract.

d

 From Smith et al. (1976).

e

 Calculated from regression equation.

f

 Not used in calculation of Species Mean Acute Value because data are available for a more sensitive life stage.
40 Draft November 12, 2004 

Table 1b. Acute Toxicity o f Selenium to Saltw ater An imals

Species Methoda Chemical

Salinity 

 (g/kg) 

LC50

or EC50

 (μg/L)b

Species Mean

Acute Value

 (μg/L) Reference

SALTWATER SPECIES

Selenite

Blue mussel 

(embryo), 

Mytilus edulis

S, U Selenium 

oxide

33.79 >10,000 >10,000 Martin et al. 1981

Bay scallop 

(juvenile),

Argopecten irradians

R, U Sodium

selenite

25 255 255 Nelson et al. 1988

Pacific oyster 

(embryo),

Crassostrea gigas

S, U Selenium

oxide

33.79 >10,000 - Glickstein 1978; 

Martin et al. 1981

Pacific oyster 

(embryo),

Crassostrea gigas

S, U Sodium

selenite

33.79 >10,000 >10,000 Glickstein 1978

Surf clam 

(juvenile),

Spisula solidissima

R, U Sodium

selenite

25 1,900 1,900 Nelson et al. 1988

Copepod 

(adult),

Acartia clausi

S, U Selenious 

acid

30 2,110 2,110 Lussier 1986

Copepod 

(adult),

Acartia tonsa

S, U Selenious 

acid

30 839 839 Lussier 1986

Mysid 

(juvenile),

Americamysis bahia

S, U Selenious

acid

- 600 - U.S. EPA 1978

Mysid 

(juvenile),

Americamysis bahia

F, M Selenious

acid

15-20 1,500 1,500 Ward et al. 1981

Brown shrimp 

(juvenile),

Penaeus aztecus

S, U Sodium

selenite

30 1,200 1,200 Ward et al. 1981

Dungeness crab 

(zoea 1arva),

Cancer magister

S, U Selenium

oxide

33.79 1,040 1,040 Glickstein 1978

Blue crab 

(juvenile),

Callinectes sapidus

S, U Sodium

selenite

30 4,600 4,600 Ward et al. 1981
Table 1b. Acute Toxicity of Selenium to Saltw ater Animals (continued).

Species Methoda Chemical

Salinity 

 (g/kg) 

LC50

or EC50

 (μg/L)b

Species Mean

Acute Value

 (μg/L) Reference

41 Draft November 12, 2004 

Haddock 

(larva),

Melanogrammus

aeglefinus

S, U Selenious 

acid

30 599 599 Cardin 1986

Sheepshead minnow

(juvenile),

Cyrinodon variegatus

S, U Selenious 

acid

- 6,700 - Heitmuller et al.

1981

Sheepshead minnow

(juvenile),

Cyrinodon variegatus

F, M Sodium

selenite

30 7,400 7,400 Ward et al. 1981

Atlantic silverside

(juvenile),

Menidia menidia

S, U Selenious

acid

30 9,725 9,725 Cardin 1986

Fourspine stickleback

(adult),

Apeltes quadracus

S, U Selenious

acid

30 17,350 17,350 Cardin 1986

Striped bass,

Morone saxatilis

S, U Sodium

selenite

1 1,550 - Palawski et al.

1985

Striped bass 

(24 d posthatch),

Morone saxatilis

S, U Sodium

selenite

5 3,400 - Chapman 1992

Striped bass 

(25 d posthatch),

Morone saxatilis

S, U Sodium

selenite

5 3,300 - Chapman 1992

Striped bass 

(31 d posthatch),

Morone saxatilis

S, U Sodium

selenite

5 3,800 - Chapman 1992

Striped bass 

(32 d posthatch),

Morone saxatilis

S, U Sodium

selenite

5 3,900 3,036 Chapman 1992

Pinfish 

(juvenile),

Lagodon rhomboides

S, U Sodium

selenite

30 4,400 4,400 Ward et al. 1981

Summer flounder

(embryo),

Paralichthys dentatus

S, U Selenious

acid

30.2 3,497 3,497 Cardin 1986

Winter flounder 

(larva),

Pseudopleuronectes

americanus

S, U Selenious

acid

30 14,240 - Cardin 1986

Winter flounder 

(larva),

Pseudopleuronectes

americanus

S, U Selenious

acid

28 15,070 14,649 Cardin 1986
Table 1b. Acute Toxicity of Selenium to Saltw ater Animals (continued).

Species Methoda Chemical

Salinity 

 (g/kg) 

LC50

or EC50

 (μg/L)b

Species Mean

Acute Value

 (μg/L) Reference

42 Draft November 12, 2004 

Selenate

Striped bass 

(24 d posthatch),

Morone saxatilis

S, U Sodium

selenate

5 26,300c- Chapman 1992

Striped bass 

(25 d posthatch),

Morone saxatilis

S, U Sodium

selenate

5 23,700c- Chapman 1992

Striped bass 

(31 d posthatch),

Morone saxatilis

S, U Sodium

selenate

5 26,300c- Chapman 1992

Striped bass 

(32 d posthatch),

Morone saxatilis

S, U Sodium

selenate

5 29,000c- Chapman 1992

Striped bass 

(juvenile),

Morone saxatilis

F, M Sodium

selenate

6.0-6.5 85,840c

- Klauda 1985a,b

Striped bass 

(prolarvae),

Morone saxatilis

F, M Sodium

selenate

3.5-4.2 9,790 9,790 Klauda 1985a,b

a

 S = static; R = renewal; F = flow-through; M = measured; U = unmeasured.

b

 Concentration of selenium, not the chemical. Note: The values underlined in this column were used to calculate the SMAV

 for the respective species.

c

 Not used in calculation of Species Mean Acute Value because data are available for a more sensitive life stage.
43 Draft November 12, 2004 

Table 2a. Ranked Freshwater Genus Mean Acute Values

Ranka

Genus Mean

Acute Value

 (μg/L) Species

Species Mean

Acute Value

 (μg/L)b

 

Number of Acute

Values used to

Calculate Species

 Mean Value b

 

FRESHWATER SPECIES

Selenite

28 203,000 Leech,

Nephelopsis obscura

203,000 1

27 42,500 Midge,

Tanytarsus dissimilis

42,500 1

26 35,356 Midge,

Chironomus decorus

48,200 1

Midge,

Chironomus plumosus

25,934 2

25 35,000 Common carp,

Cyprinu s carpio

35,000 1

24 34,914 Snail,

Aplexa hypnorum

34,914 2

23 28,500 Bluegill,

Lepomis macrochirus

28,500 1

22 26,100 Goldfish,

Carassius auratus

26,100 1

21 24,100 Snail,

Physa sp.

24,100 1

20 24,008 White sucker,

Catostomus commersoni

30,176 2

Flannelmouth sucker

Catostom us latipinnis

19,100 1

19 15,675 Arctic grayling

Thymallus arcticus

15,675 1

18 13,600 Channel catfish,

Ictalurus punctatus

13,600 1

17 12,801 Colorado squawfish,

Ptychocheilus lucias

12,801 6

16 12,600 Mosquitofish,

Gambusia affinis

12,600 1

15 11,700 Yellow perch,

Perca flavescens

11,700 1

14 11,200 Golden shiner,

Notemigonus crysoleucas

11,200 1

13 10,580 Chinook salmon,

Oncorhynchus tshawytscha

15,596 6
Table 2a. Ranked Freshwater Genus Mean Acute Values (continued)

Ranka

Genus Mean

Acute Value

 (μg/L) Species

Species Mean

Acute Value

 (μg/L)b

 

Number of Acute

Values used to

Calculate Species

 Mean Value b

 

44 Draft November 12, 2004 

Coho salmon,

Oncorhynchus kisutch

7,240 3

Rainbow trout,

Oncorhynchus mykiss

10,488 2

12 10,200 Brook trout

Salvelinu s fontinalis

10,200 1

11 9,708 Bonytail

Gilas elegans

9,708 5

10 7,710 Worm,

Tubifex tubifex

7,710 1

9 7,679 Razorback sucker,

Xyrauchen texanus

7,679 6

8 6,500 Flagfish,

Jordanella floridae

6,500 1

7 3,489 Amphipod,

Gammarus pseudolimnaeus

3,489 5

6 2,209 Fathead minnow,

Pimephales promelas

2,209 8

5 1,783 Striped bass,

Morone saxatilis

1,783 2

4 1,700 Hydra,

Hydra sp.

1,700 1

3 1,341 Cladoceran,

Daphnia magna

905.3 11

Cladoceran,

Daphnia pulex

1,987 1

2 <515.3 Cladoceran,

Cerioda phnia a ffinis

<603.6 4

Cladoceran,

Ceriodaphnia dubia 

440 1

1 461.4 Amphipod,

Hyalella azteca

461.4 5
Table 2a. Ranked Freshwater Genus Mean Acute Values (continued)

Ranka

Genus Mean

Acute Value

 (μg/L) Species

Species Mean

Acute Value

 (μg/L)b

 

Number of Acute

Values used to

Calculate Species

 Mean Value b

 

45 Draft November 12, 2004 

Selenate

 (at sulfate = 100 mg/L)

18 1,515,661 Leech,

Nephelopsis obscura

1,515,661 1

17 661,816 Snail,

Aplexa hypnorum

661,816 1

16 226,320 Channel catfish,

Ictalurus punctatus

226,320 1

15 216,033 Bluegill,

Lepomis macrochirus

216,033 1

14 94,159 Arctic grayling,

Thymallus arcticus

94,159 2

13 68,582 Midge,

Paratanytarsus

parthenogeneticus

68,582 1

12 50,727 Midge,

Chironomus decorus

50,727 1

11 47,164 Chinook salmon,

Oncorhynchus tshawytscha

83,353 5

Coho salmon,

Oncorhynchus kisutch

29,141 3

Rainbow trout,

Oncorhynchus mykiss

43,192 2

10 27,380 Flannelmouth sucker

Catostom us latipinnis

27,380 1

9 25,032 Hydra,

Hydra sp.

25,032 1

8 18,484 Colorado squawfish,

Ptychocheilus lucius

18,484 3

7 11,346 Fathead minnow,

Pimephales promelas

11,346 5

6 10,560 Bonytail,

Gila elegans

10,560 1

5 10,309 Razorback sucker,

Xyrauchen texanus

10,309 6

4 2,522 Amphipod,

Gammarus la custris

2,747 1

Amphipod,

Gammarus pseudolimnaeus

2,315 5
Table 2a. Ranked Freshwater Genus Mean Acute Values (continued)

Ranka

Genus Mean

Acute Value

 (μg/L) Species

Species Mean

Acute Value

 (μg/L)b

 

Number of Acute

Values used to

Calculate Species

 Mean Value b

 

46 Draft November 12, 2004 

3 1,887 Cladoceran,

Daphnia magna

3,314 6

Cladoceran,

Daphnia pulex

3,420 1

Cladoceran,

Daph nia pulica ria

593 1

2 1,397 Amphipod,

Hyalella azteca

1,397 4

1 842 Cladoceran,

Cerioda phnia d ubia

842 1

a

 Ranked from most resistant to most sensitive based on Genus Mean Acute Value. Inclusion of

 "greater than" and "less than" values does not necessarily imply a true ranking, but does allow

 use of all genera for which data are available so that the Final Acute Value is not unnecessarily

 lowered.

b

 From Table 1a.
Table 2b. Ranked Saltwater Genus Mean Acute Values

Ranka

Genus Mean

Acute Value

 (μg/L) Species

Species Mean

Acute Value

 (μg/L)b

 

Number of Acute

Values used to

Calculate Species

 Mean Value b

 

47 Draft November 12, 2004 

SALTWATER SPECIES

Selenite

17 17,350 Fourspine stickleback,

Apeltes quadracus

17,350 1

16 14,649 Winter flounder,

Pseudopleuronectes

americanus

14,649 2

15 >10,000 Blue mussel,

Mytilus ed ulis

>10,000 1

14 >10,000 Pacific oyster,

Crassostrea gigas

>10,000 2

13 9,725 Atlantic silverside,

Menidia menid ia

9,725 1

12 7,400 Sheepshead minnow,

Cyprinodon variegatus

7,400 1

11 4,600 Blue crab,

Callinectes sapidus

4,600 1

10 4,400 Pinfish,

Lagodon rhomboides

4,400 1

9 3,497 Summer flounder,

Paralichthys dentatus

3,497 1

8 3,036 Striped bass,

Morone saxatilis

3,036 5

7 1,900 Surf clam,

Spisula solidissima

1,900 1

6 1,500 Mysid,

Americamysis bahia

1,500 1

5 1,331 Copepod,

Acartia clausi

2,110 1

Copepod,

Acartia tonsa

839 1

4 1,200 Brown shrimp,

Penaeus aztecus

1,200 1

3 1,040 Dungeness crab,

Cancer magister

1,040 1

2 599 Haddock,

Melanogrammus aeglefinus

599 1
Table 2b. Ranked Saltwater Genus Mean Acute Values

Ranka

Genus Mean

Acute Value

 (μg/L) Species

Species Mean

Acute Value

 (μg/L)b

 

Number of Acute

Values used to

Calculate Species

 Mean Value b

 

48 Draft November 12, 2004 

1 255 Bay scallop,

Argopecten irradians

255 1

Selenate

1 9,790 Striped bass,

Morone saxatilis

9,790 1

a

 Ranked from most resistant to most sensitive based on Genus Mean Acute Value. Inclusion of

 "greater than" and "less than" values does not necessarily imply a true ranking, but does allow

 use of all genera for which data are available so that the Final Acute Value is not unnecessarily

 lowered.

b

 From Table 1b.

 

Selenite

 Fresh Water

Final Acute Value = 5 14.9 μg/L

Criterion M aximum C oncentration = (514.9 μg/L) ÷ 2 = 257 μg/L

 Salt Water

Final Acute Value = 2 53.4 μg/L

Criterion M aximum C oncentration = (253.4 μg/L) ÷ 2 = 127 μg/L

Selenate

 Fresh Water

Final Acute Value = 834.4 μg/L (calculated at a sulfate level of 100 mg/L from GMAVs)

Criterion Maximum Concentration = (834.4 μg/L) ÷ 2 = 417 μg/L ( at a sulfate level of 100 mg/L)

Pooled Slope = 0.5812 (see Appendix A)

ln (Criterion Maximum Intercept) = ln(417.2) - [slope x ln(100)]

 = 6.0335 - (0.5812 x 4.605) = 3.357 

 Criterion Maximum Concentration for Selenate (at a sulfate level of 100 mg/L) = e

(0.5812[ln(sulfate)]+3.357)
49 Draft November 12, 2004 

Table 3a. Ratios of Freshwater Species Mean Acute Values for Selenite and Selenate.

Selenite

Sensitivity

Rank from 

Table 2aa

Species

Selenite

Species Mean

Acute Value

 (μg/L)b

 

Selenate

Species Mean

Acute Value at

Sulfate = 100

 (μg/L)b

 Ratio

FRESHWATER SPECIES

28 Leech,

Nephelopsis obscura

203,000 1,515,661 0.134

27 Midge,

Tanytarsus dissimilis

42,500 NAc NA

26 Midge,

Chironomus decorus

48,200 50,727 0.95

Midge,

Chironomus plumosus

25,934 NA NA

25 Common carp,

Cyprinu s carpio

35,000 NA NA

24 Snail,

Aplexa hypnorum

34,914 616,816 0.057

23 Bluegill,

Lepomis macrochirus

28,500 216,033 0.132

22 Goldfish,

Carassius auratus

26,100 NA NA

21 Snail,

Physa sp.

24,100 NA NA

20 White sucker,

Catostomus commersoni

30,176 NA NA

Flannelmouth sucker

Catostom us latipinnis

19,100 27,380 0.698

19 Arctic grayling

Thymallus articus

15,675 94,159 0.166

18 Channel catfish,

Ictalurus punctatus

13,600 226,320 0.06

17 Colorado squawfish,

Ptychocheilus lucias

12,801 18,484 0.693

16 Mosquitofish,

Gambusia affinis

12,600 NA NA

15 Yellow perch,

Perca flavescens

11,700 NA NA

14 Golden shiner,

Notoemigonus crysoleucas

11,200 NA NA
Table 3a. Ratios of Freshwater Species Mean Acute Values for Selenite and Selenate (continued).

Selenite

Sensitivity

Rank from 

Table 2aa

Species

Selenite

Species Mean

Acute Value

 (μg/L)b 

Selenate

Species Mean

Acute Value at

Sulfate = 100

 (μg/L)b Ratio

50 Draft November 12, 2004 

13 Chinook salmon,

Oncorhynchus tshawytscha

15,596 83,353 0.187

Coho salmon,

Oncorhynchus kisutch

7,240 29,141 0.248

Rainbow trout,

Oncorhynchus mykiss

10,488 43,192 0.243

12 Brook trout

Salvelinu s fontinalis

10,200 NA NA

11 Bonytail

Gilas elegans

9,708 10,560 0.919

10 Worm,

Tubifex tubifex

7,710 NA NA

9 Razorback sucker,

Xyrauchen texanus

7,679 10,309 0.745

8 Flagfish,

Jordanella floridae

6,500 NA NA

7 Amphipod,

Gammarus pseudolimnaeus

3,489 2,315 1.507

6 Fathead minnow,

Pimephales promelas

2,209 11,346 0.195

5 Striped bass,

Morone saxatilis

1,783 NA NA

4 Hydra,

Hydra sp.

1,700 25,032 0.068

3 Cladoceran,

Daphnia magna

905.3 3,314 0.273

Cladoceran,

Daphnia pulex

1,987 3,420 0.581

2 Cladoceran,

Cerioda phnia a ffinis

<603.6 NA NA

Cladoceran,

Ceriodaphnia dubia 

440 842 0.523

1 Amphipod,

Hyalella azteca

461.4 1,397 0.33

a

 Ranked from most resistant to most sensitive based on selenite Genus Mean Acute Value (from Table 2a).

b

 From Table 1a.

c

 NA = N ot Available
51 Draft November 12, 2004 

Table 3b. Ratios of Saltwater Species Mean Acute Values for Selenite and Selenate.

Sensitivity

Rank from 

Table 2ba

Species

Selenite

Species Mean

Acute Value

 (μg/L)b

 

Selenate

Species Mean

Acute Value

 (μg/L)b

 Ratio

SALTWATER SPECIES

8 Striped bass,

Morone saxatilis

3,036 9,790 0.31

a

 Ranked from most resistant to most sensitive based on Genus Mean Acute V alue (from Table 2b).

b

 From Table 1b.
52 Draft November 12, 2004 

Figure 1. Ranked summary of selenite GMAVs (freshwater).
53 Draft November 12, 2004 

Figure 2. Ranked summary of selenate GMAVs (saltwater).
54 Draft November 12, 2004 

Figure 3. Ranked summary of selenate GMAVs (freshwater) at a sulfate level of 100 mg/L.
55 Draft November 12, 2004 

Review and Analysis of Chronic Data

Since the issuance of the 1987 chronic criterion of 5 μg/L, considerable information has come forth

regarding the route of exposure of selenium to aquatic organisms. Studies have shown that diet is the

primary route of exposure that controls chronic toxicity to fish, the group considered to be the most

sensitive to chronic selenium exposure (Coyle et al. 1993; Hamilton et al. 1990; Hermanutz et al. 1996). 

Chronic tests in which test organisms were exposed to selenium only through water and which have

measured selenium in the tissue of the test species have produced questionably low chronic values based

on the tissue concentrations. Some of these water-only exposures have required aqueous concentrations

of selenium of greater than 300 μg/L to attain body burdens sufficient to achieve a chronic response that

would have been reached in the real world at aqueous concentrations approximately 30 times lower

(Cleveland et al. 1993; Gissel-Nielsen and Gissel-Nielsen 1978). 

Because diet controls selenium chronic toxicity in the environment and water-only exposures require

unrealistic aqueous concentrations in order to elicit a chronic response, only studies in which test

organisms were exposed to selenium in their diet alone or in their diet and water were considered in the

derivation of a chronic value. To be able to use the chronic study results, the measurements had to

include selenium in the test species tissue. Both laboratory and field studies were considered in the

review process. Chronic studies reviewed were obtained through a literature search extending back to

the last revision review, from information supplied to U.S. EPA through the Notice of Data Availability,

and using the references cited in previous selenium criteria documents. 

Selection of Medium for Expressing Chronic Criterion

Whole-body tissue concentration of selenium on a dry weight basis, for species eliciting the chronic

response, was selected as the medium from which to base the chronic criterion value. As discussed

above, a water-based criterion is not appropriate for selenium because diet is the most important route of

exposure for chronic toxicity. The option of basing the chronic criterion on the concentration of

selenium in prey species (that is, in the diet of the target species), was considered inappropriate for two

reasons: 1) the concentration of selenium in the diet is an indirect measure of effects observed in the test

species and is dependent on feeding behavior of the target species, and 2) selection of what organism to

sample to assess attainment of a criterion based on diet is problematic in the implementation of such a

criterion. Sediment has also been proposed as a medium upon which to base the selenium chronic

criterion (Canton and Van Derveer 1997; Van Derveer and Canton 1997), but because of the patchiness
56 Draft November 12, 2004 

of selenium in sediment and an insufficient amount of data to support a causal link between

concentrations of selenium in sediment and chronic effects observed in fish (see Hamilton and Lemly

1999, for a review), a sediment-based criterion was rejected. 

Besides being a direct link to chronic endpoints, a tissue-based criterion has the positive attributes of

integrating many site-specific factors, such as chemical speciation and rates of transformation, large

variations in temporal concentrations in water, types of organisms constituting the food chain, and rates

of exchange between water, sediment, and organisms (Hamilton, in preparation; U.S. EPA 1998). 

Whole-body tissue was selected over specific tissue types, such as ovary, liver, kidney or muscle because

of practical reasons of sampling and because a sufficient data base containing chronic effects based on

whole-body tissue is present in the literature. Ovaries may be the best tissue to link selenium to

reproductive effects because of its role in the maternal transfer of selenium to eggs, and embryo-larval

development being one of the most sensitive endpoint for chronic effects. However, ovarian tissue is

also only available seasonally and sometimes difficult to extract in quantities sufficient for analysis,

especially in smaller fish species. Whole-body larval tissue is also not practical due to sampling and

seasonal constraints.

To increase the number of studies in which chronic effects could be compared with selenium

concentrations in whole-body tissue, the relationships between selenium concentrations in whole-body

and selenium concentrations in ovary, liver, and muscle tissues were estimated. Data from 4 dietary

exposure studies that sampled whole-body as well as muscles, ovary, or liver allowed the projection of

whole-body concentrations as a function of concentrations in these individual tissues. It was not possible

to estimate such relationship for kidneys and carcass because of insufficient data. One species (bluegill

sunfish) comprised over 90 percent of the data evaluated for these relationships. 

Median concentrations of selenium in the whole-body were projected as a linear function of selenium

concentrations in ovaries and liver, or as an exponential function of the natural logarithm of selenium

concentrations in muscles (Figure 4; Appendix H). When selenium concentration in more than one organ

or tissue was available, muscle tissue was used preferentially for converting into an equivalent wholebody value. Where appropriate, whole-body selenium concentrations were estimated from selenium

concentrations in muscle, ovary and liver according to the following equations (see Appendix H for

details on statistical analyses):
57 Draft November 12, 2004 

Figure 4. The quantile regression curves project median selenium concentrations in the whole body of

bluegill, largemouth bass, tilapia and carp as a function of selenium concentrations in their

tissues. Most data are from bluegill. Estimates of model parameters minimize the sum of

weighted absolute deviations (see Appendix H for details about statistical analyses). 
58 Draft November 12, 2004 

[Sewhole-body] = exp(0.1331 + (0.8937 × ln[Semuscle])) (I)

[Sewhole-body] = 0.0173 + (0.4634 × [Seovary]) (II)

[Sewhole-body] = -0.2609 + (0.3071 × [Seliver]) (III)

Chronic studies that reported selenium concentrations in tissues based on wet weight were converted to

dry weight using a moisture content of 0.80 (U.S. EPA 1985b), unless specified otherwise. Note that

because conversion from wet to dry weight and from tissue to whole-body selenium concentration can

increase uncertainty in the estimate, site-data analysts should develop their own conversion factors

whenever possible to improve accuracy. The basis for such factors can be obtained from local historical

data or from newly acquired data specific for that site and species.

Calculation of Chronic Values

In aquatic toxicity tests, chronic values have usually been defined as the geometric mean of the highest

concentration of a toxic substance at which no adverse effect is observed (highest no observed adverse

effect concentration, NOAEC) and the lowest concentration of the toxic substance that causes an adverse

effect (lowest observed adverse effect concentration, LOAEC). The significance of observed effects is

determined by statistical tests comparing responses of organisms exposed to natural concentrations of the

toxic substance (control) against responses of organisms exposed to elevated concentrations. Analysis of

variance is the most common test employed for such comparisons. This approach however, has its

limitations. Since neither NOAEC or LOAEC are known in advance and the number of concentrations

that can be tested is constrained by logistic and financial resources, observed effects of elevated

concentrations may not permit accurate estimates of chronic values. For instance, if all elevated

concentrations had high adverse effects or if the difference in concentrations between two significantly

different treatments was large, it would not be possible to define either the NOAEC or LOAEC with

precision. Furthermore, as the concentration of some substances (e.g., selenium) naturally varies among

ecosystems, a concentration that is above the normal range at one site, maybe within the normal range at

a different location. In this approach to calculate chronic values, natural variation in concentrations of a

substance implies that controls are site specific, and thus multiple tests are needed to define the chronic

value at different locations.
59 Draft November 12, 2004 

An alternative approach to calculate chronic values focuses on the use of regression analysis to define the

dose-response relationship. With a regression equation, which defines the level of adverse effects as a

function of increasing concentrations of the toxic substance, it is possible to determine the concentration

that causes a relatively small effect, for example a 5 to 30 percent reduction in response. A reduction of

20 percent in the response observed at control (EC20) was used as the chronic value because it represents

a low level of effect that is generally significantly different from the control (U.S. EPA 1999). Smaller

reductions in growth, survival, or other endpoints only rarely can be detected statistically. Effect

concentrations associated with such small reductions have wide uncertainty bands, making them

unreliable for criteria derivation. Adverse effects are generally modeled as a sigmoid function of

increasing concentrations of the toxic substance (Figure 5).

Figure 5. Reductions in survival, growth or other responses of organisms are often modeled as a sigmoid

function of increasing concentrations of selenium, or any other toxic substance.

A logistic regression was used to model negative effects of increasing concentrations of selenium on

growth, survival, or percent of normal individuals (without deformities) of several aquatic species. The

equations that described such functions were then used to estimate the concentration that promoted a 20

percent reduction in response observed at control levels (EC20). These analyses were performed using

the Toxic Effects Analysis Model software (version 0.02; R. Erickson, U.S. EPA Duluth).
60 Draft November 12, 2004 

Only data sets that met the following conditions were included in the analysis: (1) the experiment had a

control treatment, which made it possible to define response levels at natural concentrations of selenium,

(2) and at least four concentrations of selenium. (3) The highest tested concentration of selenium caused

>50 percent reduction relative to the control treatment, and (4) at least one tested concentration of

selenium caused <20 percent reduction relative to the control treatment to ensure that the EC20 was

bracketed by tested concentrations of selenium. When the response was expressed as percentages (e.g., 

percent survival), transformed values (arcsin of the square root) were used to homogenize the variance.

Logistic regression assumes that a logistic model describes the log dose-response curve. For a visual

display of such model, a logistic curve with three parameters was fitted to each data set using nonlinear

least-squares regression analysis (Draper and Smith 1981). The logistic model was

where x symbolizes the selenium concentration in the organism’s tissues, y is the response of interest

(survival, growth, or reproduction), and y0, a and b are model parameters estimated by the regression

analysis. The y0 parameter represents the response of interest at background levels of selenium. The

graphs also include the 95 percent confidence interval for projections of the logistic model. These tasks

were performed in S-Plus version 6.0 (Insightful 2001).

When the data from an acceptable chronic test met the conditions for of the logistic regression analysis,

the EC20 was the preferred chronic value. When data did not meet the conditions, best scientific

judgment was used to determine the chronic value. In this case the chronic value is the geometric mean

of the NOAEC and LOAEC and termed the maximum allowable toxicant concentration (MATC). But

when no treatment concentration was an NOAEC, the chronic value is less than the lowest tested

concentration. And when no treatment concentration was a LOAEC, the chronic value is greater than the

highest tested concentration.

Evaluation of Freshwater Chronic Data for Each Species

Acceptable freshwater chronic toxicity data are currently available for an aquatic invertebrate

(Brachionus calyciflorous), eight different fish species, and a mix of fish species from the family

Centrarchidae in a total of 21 distinct studies (Table 4). Detailed summaries of each study are included

in Appendix I. Collectively, only these data were considered for the derivation of a final tissue residue

criterion for selenium. Below is a brief synopsis of the experimental design, test duration, relevant test
61 Draft November 12, 2004 

endpoints, and other critical information regarding the derivation of each specific chronic value. The

chronic toxicity values for other chronic selenium toxicity values and endpoints are included in

Appendix I. 

Brachionus calyciflorus (freshwater rotifer)

This study reported by Dobbs et al. (1996) 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 μg Se/L in water, and the population survived only 6 days at selenium

concentrations equal to or greater than 202.4 μg Se/L in the water (40 μg Se/g dw in the algae). 

Regression analysis of untransformed growth data (dry weight) determined 4 day post-test initiation

resulted in a calculated EC20 of 42.36 μg Se/g dw tissue (Table 4).

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 highselenium mosquitofish replacement diet (SLD diet type) were slightly elevated compared to the

replacement diet composed of uncontaminated control mosquitofish that were spiked with organic

selenium (SeMe diet type). 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: boron- 10 μg/g; chromium- 2.8 μg/g, iron- 776 μg/g, and

strontium- 48.9 μg/g.

 
62 Draft November 12, 2004 

During the test, the survival of control chinook salmon larvae and larvae fed the lowest dietary selenium

concentrations in either dietary exposure type (SLD and SeMe, respectively, consuming food at

approximately 3 μg Se/g dw) exceeded $97 percent up to 60 days post-test initiation. Between 60 and 90

days of exposure, however, the control survival declined significantly. Therefore, only data collected up

to 60 days post-test initiation was considered for analysis. Regression analysis of untransformed growth

data after 60 days of exposure resulted in a calculated EC20 of 15.74 μg Se/g dw tissue for fish fed the

SLD diet type, and 10.47 μg Se/g dw tissue for fish fed the SeMe diet type (Table 4). Note: The

mosquitofish from San Luis Drain were not tested for contaminants other than certain key elements

suspected to be present in these fish. The San Luis Drain receives irrigation drainage from the greater

San Joaquin Valley; and therefore, there is the possibility that the mosquitofish used in this study may

have contained elevated levels of pesticides. The use of the SLD diet results assumes that selenium, and

not these other possible contaminants, was the cause of any adverse chronic effects.

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. Fish fed the diets (low carbohydrate and

high carbohydrate) with the highest selenium concentration (11.4 and 11.8 μg Se/g dw food,

respectively) exhibited a 45 to 48 percent reduction in body weight (expressed as kg per 100 fish)

compared to control fish by the end of the exposure, which the authors attributed 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 final concentrations of selenium

in liver tissue of trout reared on the high carbohydrate seleniferous dietary type is the geometric mean

(GM) of 21.0 μg Se/g dw (NOAEC) and 71.7 μg Se/g dw (LOAEC), or 38.80 μg Se/g dw. Using the

equation III to convert the selenium concentration in liver tissue to a concentration of selenium in the

whole-body, the MATC becomes 11.65 μg Se/g dw (Table 4). The calculated MATC for the same group

of experimental fish exposed to selenium in the low carbohydrate diet becomes 13.08 μg Se/g dw 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 I).

Hilton et al. (1980) employed a similar test design as Hilton and Hodson (1983) in a later experiment to

examine the narrow window at which selenium changes from an essential nutrient to a toxicant affecting
63 Draft November 12, 2004 

juvenile rainbow trout. The food consisted of a casein-torula yeast diet supplemented with selenium as

sodium selenite. 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 μg Se/g dw) 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 GM of the NOAEC (40 μg Se/g dw tissue) and the LOAEC (100 μg Se/g dw tissue), or

63.25 μg Se/g dw. Using equation III to convert selenium concentrations in the liver to selenium

concentrations in the whole body, the MATC becomes 19.16 μg/g dw (Table 4).

Eggs and milt were obtained from ripe rainbow trout collected from reference streams and streams

containing elevated selenium from an active coal mine in Alberta, Canada (Holm 2002; Holm et al.

2003). Eggs were fertilized and monitored in the laboratory until swim-up stage for percent fertilization,

deformities (craniofacial, finfold, and spinal malformations), edema, and mortality. Similar

investigations were conducted in 2000 and in 2001. The effort in 2001 added a stream with an

intermediate level of selenium contamination and another reference stream. The only other notable

difference between 2000 and 2001 was the temperature at which the embryos were incubated; 8°C in

2000 and 5°C in 2001. The author stated 5°C more closely approximated actual incubation temperatures

for rainbow trout eggs. No differences were observed for percent fertilization or mortality between the

reference and contaminated sites in both the 2000 and 2001 investigations. The frequencies of

embryonic deformities and edema were significantly greater in the stream affected by coal mining than in

the reference stream in the 2000 study. The average frequencies of embryonic craniofacial, skeletal and

finfold deformities in the contaminated stream were 7.7, 13.8 and 3.2 percent, respectively; the average

frequency of edematous embryos was 30.8 percent. The effect level for selenium was determined to be

the average selenium concentration in rainbow trout muscle tissue, 1.50 μg Se/g ww. Muscle ww was

converted to dw using 75.84 percent moisture derived for rainbow trout and equation 1 was used to

convert selenium muscle dw to selenium in whole body dw. The chronic value determined for

embryonic abnormalities in rainbow trout (2000 study) was 5.79 μg Se/g adult whole body dw. A

comparison of the frequency of embryonic deformities or edema between selenium contaminated and

reference streams with the 2001 data indicated there were no significant differences. An EC20 value,

however, was computed for the relationship between craniofacial deformities and the concentration of
64 Draft November 12, 2004 

selenium in eggs, 10.4 μg Se/g eggs ww. Quantile regression was used to convert selenium in egg ww to

muscle ww using the rainbow data reported by Holm et al. (2003). The remaining conversion to the

whole body dw value of 5.85 μg Se/g was made using 75.84 percent moisture and equation 1. See

Appendix I for details on these studies. 

Oncorhynchus clarki (cutthroat trout)

No significant effects of bioaccumulated selenium on mortalities and deformities in the eggs, larvae, and

fry from wild-caught cutthroat trout from a reference and exposed site (Fording River, British Columbia,

Canada) were observed by Kennedy et al. (2000). 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 muscle tissue from adult fish

was 2.4 μg/g dw tissue for fish collected from the reference site, and 12.5 μg/g dw tissue for fish

collected from the Fording River. Using Equation I to convert the selenium concentration in muscle

tissue to a selenium concentration in the whole-body, the chronic value for this study was estimated to be

>10.92 μg/g dw parental fish tissue (see Table 4). 

Hardy (2002) fed cutthroat trout experimental diets containing a range of selenomethionine (0-10 μg/g

dw) for 124 weeks. No significant growth or survival effects were observed in the adult fish over the 124

weeks which reached a whole body concentration of 12.5 μg/g dw selenium after 44 weeks. Embryolarval observations (percent hatch and percent deformed) were not related to whole body selenium

concentrations in the spawning females (9.37 μg/g dw) fed the selenium-laden diet for 124 weeks. The

chronic value for this study was determined to be >9.37 μg Se/g dw.

Salvelinus fontinalis (brook trout)

Spawning brook trout were collected from streams with elevated selenium contaminated by coal mining

activity and from reference streams in 2000 and again in 2001 (Holm 2002; Holm et al. 2003) . Similar

to that described for rainbow trout above, fertilized eggs were monitored in the laboratory for percent

fertilization, deformities (craniofacial, finfold, and spinal malformations), edema, and mortality. The

only abnormality observed in the embryos spawned from the brook trout collected in 2000 at the

contaminated stream that had a frequency greater than the reference stream was craniofacial deformity

(13.6 percent for the contaminated stream compared to 3.0 percent in the reference stream). The effect

level for craniofacial deformity in brook trout for the 2000 data was determined to be the average

selenium concentration in adult muscle tissue, 3.79 μg Se/g ww or 13.2 μg Se/g whole body dw using
65 Draft November 12, 2004 

conversion factors (75.84 percent moisture and equation 1). The only significant difference observed in

2001 brook trout was a greater frequency of finfold deformities in brook trout collected from Gregg

Creek (intermediate selenium levels) relative to the reference stream (4.1 percent in Gregg Creek

compared to 0.1 percent in the reference stream). The effect level for finfold deformites in the 2001

study was estimated to be the concentration of selenium in brook trout eggs from Gregg Creek, 6.88 μg

Se/g ww. Using the same conversion factors used for rainbow trout in the Holm study described above,

the chronic value in adult whole body dw is 12.4 μg Se/g. See Appendix I for more details. 

Salmonidae summary

Four of the studies with salmonids discussed above evaluated the effects of selenium directly on growth

of juvenile fish (Hamilton et al. 1990; Hilton and Hodson 1983; Hilton et al. 1980; Hicks et al. 1984),

while three of the studies evaluated the effects of selenium on embryo/larval survival and deformity

where exposure was through the parents (Hardy et al. 2002; Holm 2000; Holm et al. 2003; Kennedy et al.

2000). Of the studies based on embryo/larval survival and deformity where exposure was through the

parents, fry from hatchery brood fish were fed a selenium-spiked diet, grown to sexual maturity, and

spawned for the effects determination in the Hardy et al. study, and wild-caught adults from selenium

contaminated streams were spawned for the effects determination in the Holm studies and in the

Kennedy et al. study. Significant effects due to selenium exposure in these field exposed studies were

not observed for cutthroat trout (Hardy et al. 2002; Kennedy et al. 2000). Significant effects were

observed for rainbow trout and brook trout, albeit relatively minor effects in the latter species (Holm

2002; Holm et al. 2003). Although significant effects were not observed in the Hardy et al. and Kennedy

et al. studies, the data are meaningful with respect to the effect levels obtained for embryo-larval

development in Oncorhynchus, and thus retained for GMCV (10.66 μg Se/g dw) calculation (Table 4). 

Pimephales promelas (fathead minnows)

Chronic values for fathead minnows were derived from three laboratory-based studies and one mesocosm

study (Table 4). Two of the laboratory 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 (cultured in chambers without selenium containing algae)

or selenium-contaminated rotifers (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-
66 Draft November 12, 2004 

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 whole-body selenium concentrations ranging from 43.0 to 51.7 μg/g dw tissue in the first two

experiments (see Appendix I for conditions), but growth was not significantly reduced in larvae that had

accumulated 61.1 μg/g dw tissue in the third experiment (Table 4). The geometric mean of these three

values, 51.40 μg/g dw, was considered the chronic value for selenium for this test.

A similar test system was used by Dobbs et al. (1996), in which 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 μg Se/L in water, and they

survived only to 11 days at selenium concentrations equal to or greater than 393.0 μg/L in the water (75

μg Se/g dw in the diet, i.e., rotifers). The LOAEC for retarded growth (larval fish dry weight) in this

study was <73 μg Se/g dw tissue (Table 4).

In contrast to the above laboratory-based food chain studies, Ogle and Knight (1989) examined the

chronic effects of only 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, and 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 μg Se/g 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 μg Se/g dw did exhibit a

significant reduction in growth compared to controls (16 percent reduction), whereas no effect on growth

occurred in the fish fed 15.2 μg Se/g dw. The whole-body chronic value, as determined by the GM of the

NOAEC and the LOAEC measured at 98 days post-test initiation, was 5.961 μg/g dw tissue (Table 4).

The chronic value of 5.961 μg/g dw determined for growth after 98 days of exposure to pre-spawning

fathead minnow adults (Ogle and Knight, 1989) was approximately an order of magnitude lower than the
67 Draft November 12, 2004 

growth effects to fathead minnow observed in Bennett et al. (1986) and Dobbs et al (1996). The length

of exposure in the Ogle and Knight test was more than twice as long as either Bennett et al. or Dobbs et

al., suggesting a longer duration was needed in order to detect any growth effects from selenium. 

However, in addition to the absence of effects observed for the reproductive parameters measured,

survival of larvae hatched from parents exposed to each of the five selenium treatments (including those

in which growth was affected) was not affected. 

Other studies (Bryson et al. 1984; Bryson et al. 1985a; Coyle et al. 1993; Hermanutz et al. 1996) have

found larval deformities and larval survival to be the most sensitive endpoint to fish. This also appears

true for fathead minnows. Schultz and Hermanutz (1990) examined the effects of selenium in fathead

minnow larvae transferred from parental fish (females). The parental fathead minnows were originally

exposed to selenite which was added to artificial streams in a mesocosm study. The selenite entered the

food web which contributed to exposure from the 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 containing 10 μg Se/L. Selenium residues in the ovaries of females

from the treated stream averaged 39.27 μg/g dw. Using equation II to convert the selenium concentration

in the ovaries to a concentration of selenium in the whole-body, the chronic value for this species was

estimated to be <18.21 μg Se/g dw (Table 4).

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. 

This is a common observation also noted by Hilton and Hodson (1983) and Hilton et al. (1980) and

apparent in Coughlan and Velte (1989). Given the no observed effect to larval survival and the apparent

non-toxicological effect on growth in the Ogle and Knight study, the SMCV for fathead minnows does

not include the 5.961 μg/g dw chronic value.

Also excluded from the SMCV calculation for fathead minnows were the chronic value and LOAEC

estimated from the laboratory food-chain experiments of Bennett et al. (1986) and Dobbs et al. (1996). 

In both of these studies, the effect concentrations based on larval growth appear to be less sensitive than

the effect on larval edema and deformity observed in Schultz and Hermanutz (1990). The greater
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sensitivity of larval fathead minnows to selenium as measured by edema and deformity (lordosis) in the

Schultz and Hermanutz (1990) study is consistent with other studies using bluegill (Table 4); and thus,

the SMCV for fathead minnows of <18.21 μg/g dw was based on this endpoint.

Catostomus latipinnis (flannelmouth sucker)

Beyers and Sodergren (2001a) exposed flannelmouth sucker larvae to a range of aqueous selenate

concentrations (<1, 25.4, 50.6, 98.9, and 190.6 μg/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 μg/g dw). There were no survival or

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 μg Se/g dw.

Xyrauchen texanus (razorback sucker)

Two laboratory exposure studies have been done with the endangered razorback sucker. In the first

study, Beyers and Sodergren (2001a) exposed larval razorback suckers to the same aqueous and diet

concentrations as described above for the flannelmouth sucker. Similar to the results found for the

flannelmouth sucker, survival and growth of the razorback sucker larvae were not reduced after the 28

day exposure. The chronic value for this study based on selenium measured in the larvae at the end of

the test is > 12.9 μg Se/g dw. In a second study, Beyers and Sodergren (2001b) exposed larval razorback

suckers to a control water and three different site waters containing varying concentrations of selenium. 

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 in 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) the site water type, identified as De

Beque, showed a significant reduction in the growth of fish exposed to site water/site food relative to site

water/control food, but contained levels of selenium in the water (< 1μg/L) and food (2.10 μg/g dw)

typically lower than those that have been found to elicit effects. The chronic value for this study is > 42

μg Se/g dw based on the whole body concentration of selenium in the larval razorback suckers exposed

to North Pond site water.
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Lepomis macrochirus (bluegill sunfish)

Applicable chronic data for bluegill sunfish can be grouped according to field exposure versus laboratory

exposure. In some field studies, chronic tolerance to selenium appears to be much higher than in

laboratory studies (Bryson et al. 1985a). In the Bryson et al. (1984, 1985a) and Gillespie and Baumann

(1986) studies, the progeny of females collected from a selenium contaminated reservoir, Hyco

Reservoir, Person County, NC and artificially crossed 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 μg/L. The whole-body tissue selenium concentration in the female

parent associated with this high occurrence of mortality of hatched larvae was <43.70 μg/g dw tissue, as

reported by Bryson et al. (1985a), and <21.47 μg/g dw tissue, as reported by Gillespie and Baumann

(1986) (Table 4). 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. These chronic effect tissue values are in line with

the EC20 calculated for the occurrence of deformities among juvenile and adult fishes from the family

Centrarchidae collected from Belews Lake, NC, i.e., 44.57 μg Se/g dw (see Lemly 1993b, Table 4). 

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 fish tissues in these tests was in the

seleno-DL-cysteine-2X treatment (3.74 μg Se/g dw). Bryson et al. (1985b) also 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 μg/g dw or 5.45 μg Se/g dw whole body

using equation III for conversion. The chronic values for the juvenile bluegill test and the embryo-larval

development tests were >3.74 and >5.45 μg Se/g dw whole body, respectively.

In contrast, the chronic effects threshold for larval survival in a combination laboratory waterborne and

dietary selenium exposure (Coyle et al. 1993), or even a long-term mesocosm exposure (Hermanutz et al.

1996), occurs at concentrations approximately 3 times lower than those recorded above (Table 4). In the

Coyle et al. (1993) study, two-year old pond reared bluegill sunfish were exposed in the laboratory to a

nominal 10 μg Se/L in water (measured concentrations in respective dietary treatments ranging from 8.4

to 11 μg/L) and fed (twice daily ad libitum) Oregon moist pellets containing increasing concentrations of

seleno-L-methionine. The fish were grown under these test conditions for 140 days. Spawning
70 Draft November 12, 2004 

frequency, fecundity, and percentage hatch were monitored after 60 days when spawning began to occur. 

There was no effect of the combination of the highest dietary selenium concentration (33.3 μg Se/g dw)

in conjunction with waterborne selenium concentrations averaging 11 μg/L on adult growth, condition

factor, gonadal somatic index, or the various reproductive endpoints (Appendix I). The survival of newly

hatched larvae, however, was markedly reduced; only about 7 percent survived to 5 days post-hatch. 

Regression analysis on arcsin square root transformed fry survival data 5 days post-hatch resulted in a

calculated EC20 of 8.954 μg Se/g dw tissue (Table 4). 

Hermanutz et al. (1996), as corrected by Tao et al. (1999), and peer reviewed in Versar (2000), exposed

bluegill sunfish to sodium selenite spiked into artificial streams (nominal test concentrations: 0, 2.5, 10,

and 30 μg Se/L) which entered the food web, thus providing a simulated field-type exposure (waterborne

and dietary selenium exposure). A series of three studies were conducted over a 3 year period lasting

anywhere from 8 to 11 months. All three studies began exposure to adult bluegill sunfish in the fall and

ended the respective study in the summer of the following year. Winter temperatures averaged 4.1 and

4.5°C and spawning months (June-July) averaged 23.9 and 22.4°C, respectively for Studies II and III.

The Hermanutz et al. (1996) report contains the data presented in the Hermanutz et al. (1992) article

(Study I, 10 and 30 μg/L exposures) as well as Studies II and III (2.5 and 10 μg/L and recovering

mesocosms). 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 in 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 μg/L) survived. Incidence of edema, hemorrhage, and lordosis in

the larvae incubated in egg cups and spawned from fish exposed to 10 μg Se/L were 100, 45 and 15

percent, respectively (see Hermanutz 1996 in Appendix I). Such health problems were not observed in

larvae from fish that were not exposed to elevated concentrations of selenium (control treatment). Rates

of edema, hemorrhage, and lordosis occurrence in larvae (egg cup data) from fish exposed to 2.5 μg Se/L

were 0, 3.6 and 0 percent, respectively. Mean concentrations of selenium in fish tissues (whole body) of

the control, 2.5 and 10 μg Se/L treatments were 1.95, 5.55, and 26.46 μg Se/g dw, respectively. Except

for the 2.5 μg Se/L treatment, each value is a geometric mean of 2 replicates. 

Results of this experiment were not suitable for regression analysis. Exposure of adult fish to 10 μg Se/L

caused a small reduction in larval survival (in their first three days), from 75 to 57 percent. However,

responses lower than half of the values observed in control treatments are needed to adequately

characterize the slope of decline in survival (or growth, reproduction...) with increasing concentrations of
71 Draft November 12, 2004 

selenium. It is not sufficient to have only extremely low and high responses. Intermediate effects are

necessary to properly estimate the shape of the dose-response curve. The percent of larvae with edema

increased from 0 percent at the control and 2.5 μg Se/L treatments to 100 percent in streams that received

10 μg Se/L. With these data, it is not possible to accurately estimate the lowest concentration with

adverse effects (LOAEC) nor the rate at which incidence of edema increases with higher tissue

concentrations of selenium.

The chronic value for this study was estimated from results of analysis of variance (ANOVA) reported by

Tao et al. (1999). ANOVA was utilized to evaluate effects of elevated concentrations of selenium on

percent hatch, percent survival, maximum percent edema, lordosis, and hemorrhage, and minimum

percent healthy (egg cup data). Treatment effects were only significant for maximum percent edema and

minimum percent healthy (see their Table 4-19), and in no instance were differences between the 2.5 μg

Se/L and control treatments significant (Dunnett’s Means test, all probabilities > 0.1, see their Table 4-

20). These results clearly suggest that the 2.5 μg Se/L treatment had no adverse impact on bluegill

larvae. They are further supported by analysis of the field nest data (see Hermanutz 1996 in Appendix I). 

In this experiment, treatment had a significant effect on maximum percent edema (raw data and ranks)

and maximum percent hemorrhage (ranks only). Probabilities of differences between the 2.5 μg Se/L and

control treatments were >0.2 for all response variables except maximum percent hemorrhage, which had

an estimated probability of 0.05 (raw data, P=0.022 for ranks; Dunnett’s means test). Such values,

though, were well above the adjusted experiment-wise error rate for multiple comparisons ("’=0.0085,

"’=1-(1-")

1/k

; "=0.05, k=6 comparisons; Sokal and Rohlf 1981), which takes into account the fact that

selenium effects were tested on six different response variables. Therefore, the chronic value for this

study, 12.12 μg Se/g dry weight, was calculated as the geometric mean of tissue concentrations of

selenium in the 2.5 (NOAEC) and 10 μg Se/L (LOAEC) treatments (5.55 and 26.46 μg/g dw,

respectively).

The importance of diet in the bioaccumulation of selenium was demonstrated in one additional

experiment. Study III consisted of the addition of new adult bluegill to the same streams that received

the 2.5, 10 and 30 μg/L sodium selenite during previous studies, but with all dosing of selenite halted. 

The adult bluegills exposed only to dietary selenium present in the food web accumulated selenium to

levels very near to the levels 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
72 Draft November 12, 2004 

from fish that accumulated 11.7 and 14.5 μg/g dw in the recovering 10 μg/L streams, and 17.35 μg/g dw

in the recovering 30 μg/L stream. The lack of any effect on the Study III larvae suggests that although

dietary exposure would have been the predominant exposure route in both Study II and Study III ,

environmental differences influenced the toxicological significance of the tissue concentrations.

A 90-day diet-only laboratory exposure in which juvenile bluegill sunfish were fed a range of

selenomethionine concentrations added to Oregon moist did not have any significant effects on survival

(Cleveland et al. 1993). The authors did report a significant decrease in the condition factor (K) at the

diet treatment where bluegill whole body tissue concentrations were measured at 7.7 μg Se/g dw. The

condition factor (weight x 105

/length3

) is reflective of the weight of the fish, and as discussed earlier, the

avoidance of food at similar dietary concentrations in other fish studies (Ogle and Knight 1989; Hilton

and Hodson 1983; Hilton et al. 1989; Coughlan and Velte 1989) suggests the reduction in K is possibly a

non-toxicological effect. Given the very slight reduction in K (1.3 to 1.2) and the uncertain relevance of

growth data, the chronic value for this study was estimated at > 13.4 μg Se/g dw.

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 authors exposed

juvenile bluegill sunfish in the laboratory to waterborne (1:1 selenite:selenate; nominal 5 μg Se/L) and

foodborne (seleno-L-methionine in TetraMin; nominal 5 μg Se/g dw food) selenium for 180 days. Tests

with a control and treated fish were run at 4OC and 20OC with biological and selenium measurements

made every 60 days. Survival and whole-body lipid content were unaffected at 20OC (whole-body

selenium concentrations equal to 6 μg/g dw) when compared to control fish. Fish exposed to the

combination low-level waterborne and dietary selenium at 4OC 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 all indicative of an additional stress load. The chronic value for

juvenile bluegill sunfish exposed to waterborne and dietary selenium at 4OC was <7.9 μg/g dw tissue,

whereas the chronic value for juvenile bluegill sunfish exposed to waterborne and dietary selenium based

on survival at 20OC was >6 μg/g dw whole-body tissue.

Five of the studies discussed above evaluated the effects of selenium on fish larvae to which exposure

was through the parents. Three of these studies collected adult fish from Hyco Reservoir to which the

bluegill population had been exposed to elevated selenium concentrations for multiple generations

(Bryson et al. 1984; Bryson et al. 1985a; Gillespie and Baumann 1986), whereas the other two studies
73 Draft November 12, 2004 

exposed bluegill parents obtained from an uncontaminated source (Coyle et al. 1993; Hermanutz et al.

1996). The average of the chronic values reported for the Hyco studies was four times greater than the

value in the latter two studies. This difference may simply be the inability of the field tests to detect a

lower effect concentration than that which was observed at the site. However, Bryson et al. (1985a)

found no effects to larval survival from Hyco Reservoir females collected in an “unaffected area”

containing 19.18 μg Se/g dw suggesting the possibility of tolerance through physiological or genetic

adaptation of the previous exposed bluegill population at Hyco Reservoir. 

Acquisition of tolerance to selenium has also been implied in the literature for other fish species. For

example, Kennedy et al. (2000) suggested tolerance at the cellular level as an explanation for the normal

development of early life stages for cutthroat trout collected from a stream containing 13.3 to 14.5 μg

Se/L in the water column. These authors reported that the overall frequency of larval deformities in the

exposed population was less than 1 percent, and in one fish containing eggs with 81.3 μg Se/g dw, there

were 0.04 percent pre-ponding deformities and 3.3 percent larval mortalities. It should be noted that the

acquisition of tolerance to selenium has been hypothesized (Kennedy et al. 2000), but has not yet been

substantiated. Other than the Kennedy et al. study, tolerance to selenium in one of the endpoints

consistently sensitive to fish (embryo-larval development) has not been reported in the literature and its

reality is uncertain at this time. However, given the need to protect sensitive populations of species, the

chronic values for the studies in which eggs and larvae were obtained from bluegill adults that were

exposed to elevated selenium for multiple generations (i.e., Bryson et al. 1984; Bryson et al. 1985a;

Gillespie and Baumann, 1986) were not included in the SMCV calculation. 

Morone saxitilis (Striped bass)

The only remaining applicable 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 μg Se/g dw tissue) from Belews Lake, NC (treated fish) or

golden shiners with low levels of selenium (1.3 μg/g dw tissue) 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

histological damage, all eventually leading to the death of the animal. The final selenium concentration

in muscle of treated striped bass averaged from 17.50 to 20.00 μg/g dw tissue (assuming 80 percent

moisture content), which was 3.2 to 3.6 times higher than the final selenium concentrations in control
74 Draft November 12, 2004 

striped bass, which averaged 5.500 μg/g dw tissue. Using equation I to convert the selenium

concentration in muscle tissue to a selenium concentration in the whole-body, the chronic value for this

species was determined to be <14.75 μg/g dw (Table 4). 

Formulation of the Final Chronic Value (FCV) for Selenium

The lowest GMCV in Table 4 is for bluegill, 9.500 μg/g dw whole body, which is the geometric mean of

chronic values from the laboratory study of Coyle et al. (1993), the laboratory study of Lemly (1993a)

and the macrocosm exposure study of Hermanutz et al. (1996). Several of the chronic values listed in

Table 4 were not used in the calculation of this GMCV. These values fall under several categories. The

“less than” values tabulated for Bryson et al. (1984) and Gillespie and Baumann (1986) for Hyco

Reservoir bluegill were not used to because they only indicate a chronic value in a range that includes

9.500 μg/g dw. The “greater than” values for Bryson et al (1985b) were not used because similar studies

with bluegill sunfish provided more meaningful information on effect levels. The “greater than” value

for the recovering systems in Study III from Hermanutz et al. (1996) was not used in the mean calculation

because, as previously discussed in the Lepomis section, less tolerance was observed in the freshly

exposed systems of Study II. The Table 4 results for Bryson et al. (1985a) and Lemly (1993b) were also

not used in calculating the bluegill GMCV. Bryson et al. (1985a) indicated a chronic value for Hyco

Reservoir bluegill somewhere between 20.29 and 43.70 μg/g dw. Lemly (1993b), appearing in Table 4

under the category Centrarchidae, the family to which bluegill belong, yielded a chronic EC20 of 44.57

μg/g dw specific for fish from Belews Lake, NC, again substantially above the GMCV of 9.500 μg/g dw. 

It is not known whether historical exposure to elevated selenium concentrations, such as occurred at

Belews Lake and Hyco Reservoir, will dependably lead to this magnitude of increase in the chronic

tolerance of resident fish.

The Lemly (1993a) laboratory results, indicating a chronic value for over-wintering juvenile bluegill

sunfish of <7.91 μg/g dw, are not completely comparable to the other values used to calculate the bluegill

GMCV. This study involved an additional natural stress, exposure to a simulated winter low temperature

of 4°C. In this study, juvenile bluegill sunfish exposed to the over-wintering temperature 4°C appeared

to accumulate more selenium in whole-body tissues (7-8 μg Se/g dw tissue) relative to those exposed at

20°C (5-6 μg Se/g dw tissue), but also exhibited increased signs of chronic toxicity. Because this stress

occurs annually to one degree or another in nearly all the country, the FCV was lowered to 7.91 μg/g dw

to protect sensitive fish species Although the literature contains little information on the temperaturedependence of selenium toxicity, Lemly’s study (further summarized in Appendix I) was judged to be
75 Draft November 12, 2004 

sufficiently definitive to merit lowering the FCV. The study showed a clear effect on juvenile bluegill

survivorship when tissue concentrations reach 7.91 μg Se/g dw under extended cold temperature

conditions.

In the Lemly (1993a) study, the author relates the selenium induced hematological changes to gill

lamellar damage (possible reasons cited were the collection of cell parts in capillaries restricting blood

flow increasing pressure and rupturing or swelling lamellar vessels, and smaller red cells becoming

tightly packed in vessels). The author postulates that an imbalance between respiratory demands (i.e.,

Se-exposed fish used more O2

 at both 4°C and 20°C) and decreased respiratory capacity could have

constituted a stress that resulted in reduced body condition and lipid content of fish in the cold treatment. 

The condition of the combination of selenium-induced elevation in energy demand and reductions in

feeding due to cold temperature and short photoperiod, leading to severe depletion of stored body lipid

was termed, Winter Stress Syndrome. 

The Guidelines indicate that the chronic criterion (in this case the FCV) is intended to be a good estimate

of the threshold for unacceptable effect. The Guidelines point out that the threshold for unacceptable

effect does not equate with a threshold for any adverse effect. For example, some adverse effects,

possibly even a small reduction in survival, growth, or reproduction may occur at this threshold. If overwintering bluegill are as sensitive as indicated by the Lemly (1993a) results, some reduction in survival

(compared to populations accumulating lesser concentrations of selenium or exposed to less severe

winter temperatures) would occur at the FCV. Nevertheless, other studies, those of Lemly (1993b) and

Bryson et al. (1985a), suggest that historically exposed populations may not be as sensitive as the

organisms studied by Lemly (1993a). 

The bluegill exposed to selenium at 4°C in the Lemly (1993a) study accumulated 7.91 μg/g dw, whereas

those exposed to Se at 20°C accumulated only 5.74 μg/g dw. The increase in the concentration of

selenium in whole body tissue at 4°C was apparently due to reductions in lipid and body weight caused

by decreased feeding by the juvenile bluegill resulting in a concentration of selenium in their tissues. If

this concentration of selenium in tissues occurs in sensitive overwintering fish in nature, a criterion of

5.85 μg/g dw (the selenium tissue concentration in the 4°C exposure after 60 days) in for fish collected

during the summer or fall months might be warranted to protect the selenium-sensitive fish during the

winter months. However, it is not understood at this time whether fish in nature do concentrate selenium

during the winter. The Lemly (1993a) study used an artificial diet spiked with seleno-methionine. 
76 Draft November 12, 2004 

Although the 20°C fish did not show signs of food avoidance to the Se-spiked food, as discussed earlier

in this section, other studies did observe decreased feeding and effects on growth. 

If sensitive juvenile fish are indeed adversely affected during winter months, field studies should indicate

an altered age structure relative to selenium whole body tissue levels. May et al. (2001) reported that an

analysis of the size structure of bluegill populations in the Republican River and in 7 reservoirs within

this river’s basin, where mean tissue concentrations ranged from 2.85 to 8.84 mg Se/g dw, revealed large

numbers of small fishes. Similar patterns in the size structure of fish populations were observed for 7

additional species: common carp, green sunfish, channel catfish, largemouth bass, gizzard shad, black

bullhead, and river carpsucker. 

Given the uncertainty of juvenile fish concentrating selenium over the winter, an FCV of 7.91 μg Se/g

dw is recommended. However, if the concentration of selenium in whole body fish tissues approaches

5.85 μg Se/g dw during the summer or fall months, it is recommended fish be sampled during the winter

to determine if they exceed the FCV of 7.91 μg Se/g dw.

The FCV may not necessarily protect fish in a hypothetical environment where they are exposed only via

water and not via diet. If the organisms are provided with an uncontaminated diet, then exceedingly high

water concentrations, possibly above the acute criterion, are needed to elicit effects, but such effects may

occur at tissue concentrations below the FCV (Cleveland et al. 1993; Gissel-Nielsen and Gissel-Nielsen

1978). This is not a practical limitation, however, since water-only exposure of selenium is not

representative of the actual exposure of selenium to aquatic organisms in the environment. 

The FCV of 7.91 μg/g dw was based on a scientific interpretation of the data presented in Table 4. 

Although the FCV is derived from a limited number of species (9 species/7 genera), it is intended to be

protective of aquatic organisms across the United States. There may be aquatic communities whose fish

assemblage may contain species with different sensitivities to selenium compared to those listed in

Table 4. Furthermore, even within the Table 4 bluegill data, there is a range of reported tissue NOAECs

from various sites. Consequently, results from appropriate site-specific studies could be used to modify

the criterion.
77 Draft November 12, 2004 

A comparison of the FCV to tissue values measured in U.S. Fish and Wildlife Service’s National

Contaminant Biomonitoring Program and U.S. Geological Survey’s National Water Quality Assessment

(NAWQA) program is provided in Appendix J.
78 Draft November 12, 2004 

Table 4. Freshwater Chronic Values from Acceptable Tests

Species Reference Exposure route Selenium form Toxicological endpoint

Chronic value, 

μg/g dwa

SMCV

μg/g dw

GMCV

μg/g dw

Brachionus

calyciflorus

rotifer

Dobbs et al. 1996 dietary and

waterborne

(lab)

algae exposed to SeVI

in water, algae then

fed to rotifers

EC20 for rotifer dry

weight after 4 d

42.36 42.36 42.36

Oncorhynchus

tshawytscha

chinook salmon

Hamilton et al.

1990

dietary

(lab)

Se-laden mosquitofish

from San Luis Drain,

CA

EC20 for juvenile gro wth 15.74

(juvenile tissue)

12.84

10.66

Oncorhynchus

tshawytscha

chinook salmon

Hamilton et al.

1990

dietary

(lab)

Mosquitofish spiked

with seleno-DLmethionine

EC20 for juvenile gro wth 10.47

(juvenile tissue)

Oncorhynchus

mykiss

rainbow trout

Hilton and

Hodson 1983:

Hicks et al. 1984

dietary

(lab)

sodium selenite in

food preparation

MAT C for juvenile

growth;

nephrocalcinosis

11.65b

(juvenile tissue)

9.32

Oncorhynchus

mykiss

rainbow trout

Hilton et al. 1980 dietary

(lab)

sodium selenite in

food preparation

MAT C for juvenile

survival and growth

19.16b

(juvenile tissue)

Oncorhynchus

mykiss

rainbow trout

Holm 2000;

Holm et al.

2003

dietary and

waterborne

(field Luscar

River, Alberta)

not determined 2000 study: chronic

value for embryo larval

deformities

2001 study: EC20 for

craniofacial deformities

5.79c

(parent tissue)

5.85c

(parent tissue)

Oncorhynchus

clarki

cutthroat trout

Kenned y et al.

2000

dietary and

waterborne (field

- Fording River,

BC)

not determined NOAEC for

embryo/larval

deformities and

mortality

>10.92c

(parent tissue)

>10.12

Oncorhynchus

clarki

cutthroat trout

Hardy, R.W. 2002 dietary

(lab)

selenomethionine in

food preparation

NOAEC for

embryo/larval

deformities

>9.37

(parent tissue)
Species Reference Exposure route Selenium form Toxicological endpoint

Chronic value, 

μg/g dwa

SMCV

μg/g dw

GMCV

μg/g dw

79 Draft November 12, 2004 

Salvelinus

fontinalis

brook trout

Holm 2002;

Holm et al.

2003

dietary and

waterborne

(field Luscar

River, Alberta)

not determined 2000 study: chronic

value for craniofacial

deformities

2001 study: chronic

value for finfold

deformities

13.2c

(parent tissue)

12.4c

(parent tissue)

12.8 12.8

Pimephales

promelas

fathead minnow

Bennett et al.

1986

dietary

(lab)

algae expo sed to

selenite then fed to

rotifers which were

fed to fish

Chronic value for larval

growth

 51.40d

(larval tissue)

<18.21 <18.21

Pimephales

promelas

fathead minnow

Ogle and Knight 

1989

dietary

(lab)

mix of 25, 50, and 25

percent selenate,

selenite, and seleno-Lmethionine in food

preparation

MATC for pre-spawning

adult growth

5.961d

(pre-spawning adult

tissue)

Pimephales

promelas

fathead minnow

Dobbs et al. 1996 dietary and

waterborne

(lab)

algae expo sed to

selenate in water then

fed to rotifers which

were fed to fish

LOAEC for larval fish

dry weight after 8 d

<73d

(larval tissue)

Pimephales

promelas

fathead minnow

Schultz and

Hermanutz 1990

dietary and

waterborne

(mesocosm -

Monticello)

selenite added to

artificial streams

which entered food

web and provided

dietary exposure

LOAEC for larval

edema and lordosis

<18.21e

(parent tissue)

Catostomus

latipinnis

flannelmouth

sucker

Beyers and

Sodegren 2001a

dietary and

waterborne (lab)

water: selenate; 

diet: algae exp osed to

selenate in water then

fed to rotifers which

were fed to fish

NOAEC for survival and

growth

>10.2

(larval tissue)

>10.2 >10.2

Xyrauchen

texanus razorback

sucker

Beyers and

Sodegren 2001a

dietary and

waterborne (lab)

water: selenate; 

diet: algae exp osed to

selenate in water then

fed to rotifers which

were fed to fish

NOAEC for survival and

growth

>12.9 

(larval tissue)

>23.28 >23.28
Species Reference Exposure route Selenium form Toxicological endpoint

Chronic value, 

μg/g dwa

SMCV

μg/g dw

GMCV

μg/g dw

80 Draft November 12, 2004 

Xyrauchen

texanus razorback

sucker

Beyers and

Sodegren 2001b

dietary and

waterborne (lab)

water: site waters; 

diet: algae exp osed to

site water then fed to

rotifers which were

fed to fish

NOAEC for survival and

growth

>42 

(larval tissue)

Lepom is

macrochirus

bluegill

Bryson et al. 1984 dietary and

waterborne (field

- Hyco

Reservoir, NC)

not determined LOAEC for larval

mortality

<59.92c,d

(parent tissue)

9.50 9.50

Lepom is

macrochirus

bluegill

Bryson et al.

1985a

dietary and

waterborne (field

- Hyco Reservoir,

NC)

not determined Chronic value for swimup larvae<43.70c,d

 >20.29c,d

(parent tissue)

Lepom is

macrochirus

bluegill

Bryson et al.

1985b

dietary and

waterborne (field

- Hyco Reservoir,

NC)

not determined NOAEC for swim-up

larvae

 >5.45c,d

(parent tissue)

Lepom is

macrochirus

bluegill

Gillespie and

Baumann 1986

dietary and

waterborne (field

- Hyco Reservoir,

NC)

not determined Chronic value for larval

survival

<28.20d

(larval tissue); or

<21.47d,e

(parent tissue)

Lepom is

macrochirus

bluegill

Coyle et al. 1993 dietary and

waterborne (lab)

diet: seleno-Lmethionine

water: 6:1

selenate:selenite

EC20 for larval survival 8.954

(parent tissue -

females only)

Lepom is

macrochirus

bluegill

Lemly 1993a dietary and

waterborne (lab)

diet: seleno-Lmethionine

water: 1:1

selenate:selenite

LOAE C for juvenile

mortality at 4oC

<7.91

(juvenile tissue)

Lepom is

macrochirus

bluegill

Lemly 1993a dietary and

waterborne (lab)

diet: seleno-Lmethionine

water: 1:1

selenate:selenite

NOAEC for juvenile

mortality at 20oC

>6.0d

(juvenile tissue)
Species Reference Exposure route Selenium form Toxicological endpoint

Chronic value, 

μg/g dwa

SMCV

μg/g dw

GMCV

μg/g dw

81 Draft November 12, 2004 

Lepom is

macrochirus

bluegill

Hermanutz et al.

1996

dietary and

waterborne

(mesocosm -

Monticello)

selenite added to

artificial streams

which entered food

web and provided

dietary exposure

MATC for larval

survival, edem a, lordosis

and hemorrhaging Study

II

12.12

(parent tissue)

Lepom is

macrochirus

bluegill

Bryson et al.

1985b

dietary seleno-DL-cysteine NOAEC for juvenile

growth

 >3.74d

(juvenile tissue)

Lepom is

macrochirus

bluegill

Cleveland et al.

1993

dietary seleno-L-methionine NOAEC for juvenile

survival 

>13.4 d

(juvenile tissue)

Lepom is

macrochirus

bluegill

Hermanutz et al.

1996

dietary

(mesocosm -

Monticello)

selenite originally

added to artificial

streams which entered

food web and

provided dietary

exposure

NOAEC for larval

survival, edem a, lordosis

and hemorrhaging Study

III

>17.35d

(parent tissue)

Centrarchidae

(9 species)

Lemly 1993b dietary and

waterborne (field

- Belews Lake,

NC)

not determined EC20 for deformities

among juveniles and

adults

44.57

(juvenile and adult

tissue)

NA NA

Morone saxitilis

striped bass

Coughlan and

Velte 1989

dietary

(lab)

Se-laden shiners from

Belews Lake, NC

LOAEC for survival of

yearling bass

<14.75c

(juvenile tissue)

<14.75 <14.75

a All chronic values reported in this table are based o n the measured or estima ted (see footnotes below) concentration of selenium in whole body tissue. 

b Estimated using the equation III.

c Estimated using the equation I.

d Chronic value not used in SMCV calculation (see text).

e Estimated using the equation II.
82 Draft November 12, 2004 

National Criteria

The available data for selenium, evaluated in accordance with EPA’s guidelines for deriving aquatic life

criteria (Stephan et al. 1985) indicate that, except possibly where an unusually sensitive species is

important at a site, freshwater aquatic life should be protected if the following conditions are satisfied.

A. The concentration of selenium in whole-body fish tissue does not exceed 7.91 μg/g dw (dry weight). 

This is the chronic exposure criterion. In addition, if whole-body fish tissue concentrations exceed

5.85 μg/g dw during summer or fall, fish tissue should be monitored during the winter to determine

whether the selenium concentration exceeds 7.91 μg/g dw.

B. The 24-hour average concentration of total recoverable selenium in water seldom (e.g., not more than

once in three years) exceeds 258 μg/L for selenite, and likewise seldom exceeds the numerical value

given by exp(0.5812[ln(sulfate)]+3.357) for selenate. These are the acute exposure criteria. At an

example sulfate concentration of 100 mg/L, the 24-hour average selenate concentration should not

exceed 417 μg/L.

The available data for selenium, evaluated as above, indicate that saltwater aquatic life should likewise

be protected from acute effects of selenium if the 24-hour average concentration of selenite seldom

exceeds 127 μg/L. Because selenium might be as chronically toxic to saltwater fishes as it is to

freshwater fishes, the status of the fish community should be monitored if selenium exceeds 5.85 μg/g dw

in summer or fall or 7.91 μg/g dw during any season in the whole-body tissue of salt water fishes.

Implementation

As discussed in the Water Quality Standards Regulation (U.S. EPA 1983b), a water quality criterion for

aquatic life has regulatory force only after it as been adopted in a State water quality standard. Such a

standard specifies a criterion for a pollutant that is consistent with a particular designated use. With the

concurrence of the U.S. EPA, States designate one or more uses for each body of water or segment

thereof and adopt criteria that are consistent with the uses (U.S. EPA 1983c, 1987b). In each standard, a

State may adopt the national criterion (if one exists), or an adequately justified state-specific or sitespecific criterion.

Criterion concentrations, durations of averaging periods, and frequencies of allowed excursions may be

established on a state-specific or site-specific basis (U.S. EPA 1983c, U.S. EPA 1985c). Because the
83 Draft November 12, 2004 

chronic criterion is tissue-based for selenium, the averaging period only applies to the acute criterion,

which is defined as a 24-hour average, based on the speed at which effects may occur in the toxicity tests

used for its derivation. Implementation guidance on using criteria to derive water quality-based effluent

limits is available in U.S. EPA (1985c and 1987b).
A-1 Draft November 12, 2004

APPENDIX A

INFORMATION USED IN THE SULFATE CORRECTION OF 

SELENATE ACUTE TOXICITY
A-2 Draft November 12, 2004

Table A-1. Degrees of freedom (df), coefficients of determination (r

2

), slopes and respective

confidence intervals (CI) for regressions of the natural logarithm of selenate LC50% on the

natural logarithm of sulfate concentration. The “Common regression” combines regression

lines for individual species into a single model (Zar 1999), its slope is computed as in

analysis of covariance. The “Total regression” estimates a linear function for all points,

irrespective of taxa.

Species df r

2 Slope 95% CI

Fathead Minnow 14 0.83 0.48 [0.35, 0.60]

Chinook Salmon 3 0.87 0.49 [0.14, 0.83]

Gammarus pseudolimnaeus 5 0.61 0.86 [0.07, 1.66]

Hyalella azteca 4 0.39 0.19 [-0.14, 0.51]

Daphnia magna 4 0.92 0.87 [0.52, 1.22]

Ceriodaphnia dubia 11 0.84 0.70 [0.50, 0.91]

“Common regression” 46 0.65 0.58 [0.45, 0.71]

“Total regression” 51 0.36 0.76 [0.48, 1.04]
A-3 Draft November 12, 2004

Table A-2. Data used in the regressions of the natural logarithm of selenate acute values on the

natural logarithm of sulfate concentrations.

Species Age/ Size Data Source [Sulfate] 

(μg/L)

LC50 or

EC50

(μg/L) 

Adjusted

LC50

Hydra 

Hydra sp. adult Brooke et al. 1985 12000 7,300 25031.02

Leech 

Nephelopsis obscura adult Brooke et al. 1985 12000 442,000 1515577

Snail

Aplexa hypnorum Brooke et al. 1985 12000 193,000 661779.1

Cladoceran 

Ceriodaphnia dubia <24 hr Brix et al. 2001a,b 52000 1,969 2879.368

Ceriodaphnia dubia Brix et al. 2001a,b 55000 1,864 2638.398

Ceriodaphnia dubia GLEC 1999 25000 376 841.5682

Cladoceran 

Daphnia magna Dunbar et al. 1983 163000 5,300 3989.863

Daphnia magna Boyum 1984 22000 1,010 2434.939

Daphnia magna Brooke et al. 1985 12000 570 1954.477

Daphnia magna Ingersoll et al. 1990 41000 2,560 4298.133

Daphnia magna Ingersoll et al. 1990 68000 4,070 5092.556

Daphnia magna Maier et al. 1993 82000 2,840 3187.186

Daphnia pulex <24 hr Brix et al. 2001a,b 54000 10,123 14482.21

Daphnia pulex Brix et al. 2001a,b 38000 8,111 14232.89

Daphnia pulex GLEC 1999 25000 1,528 3419.99

Daphnia pulicaria Boyum 1984 22000 246 593.0643

Amphipod 

Gammarus lacustris 8-12 mm Brix et al. 2001a,b 120000 3,054 2746.951

Gammarus pseudolimnaeus adult Brooke et al. 1985 12000 75 257.168

Gammarus pseudolimnaeus Brooke 1987 12000 57 195.4477

Gammarus pseudolimnaeus GLEC 1998 25000 1,180 2641.092
Species Age/ Size Data Source [Sulfate] 

(μg/L)

LC50 or

EC50

(μg/L) 

Adjusted

LC50

A-4 Draft November 12, 2004

Gammarus pseudolimnaeus GLEC 1998 125000 2,870 2520.927

Gammarus pseudolimnaeus GLEC 1998 367000 3,710 1742.628

Gammarus pseudolimnaeus GLEC 1998 635000 3,270 1116.855

Gammarus pseudolimnaeus GLEC 1999 25000 2,191 4903.925

Amphipod 

Hyalella azteca Adams 1976 - 760 -

Hyalella azteca Brasher and Ogle 1993 13000 1,031 3374.516

Hyalella azteca Brix et al. 2001a,b 55000 1,428 2021.262

Hyalella azteca GLEC 1998 40000 2,480 4224.001

Hyalella azteca GLEC 1998 125000 1,350 1185.802

Hyalella azteca GLEC 1998 367000 1,540 723.3552

Hyalella azteca GLEC 1998 822000 3,580 1052.407

Midge

Chironomus decorus 4th instar Maier and Knight 1993 27000 23,700 50725.32

Midge

Paratanytarsus parthenogeneticus 3rd instar Brooke et al. 1985 12000 20,000 68578.14

Coho salmon 

Oncorhynchus kisutch 0.5 g Hamilton and Buhl 1990b 185000 32,500 22730.56

Oncorhynchus kisutch 1.7 g Hamilton and Buhl 1990b 291000 39,000 20963.42

Oncorhynchus kisutch alevin Buhl and Hamilton 1991 41000 158,422 265983.9

Oncorhynchus kisutch juvenile Buhl and Hamilton 1991 41000 30,932 51933.53

Oncorhynchus tshawytscha (0.7 g Hamilton and Buhl 1990b 185000 121,000 84627.63

Oncorhynchus tshawytscha 0.5 g Hamilton and Buhl 1990b 185000 100,000 69940.19

Oncorhynchus tshawytscha 1.6 g Hamilton and Buhl 1990b 291000 180,000 96754.25

Oncorhynchus tshawytscha Hamilton and Buhl 1990b 291000 134,000 72028.17

Oncorhynchus tshawytscha eyed egg Hamilton and Buhl 1990b 47000>552,000 -

Oncorhynchus tshawytscha alevin Hamilton and Buhl 1990b 47000>176,640 -
Species Age/ Size Data Source [Sulfate] 

(μg/L)

LC50 or

EC50

(μg/L) 

Adjusted

LC50

A-5 Draft November 12, 2004

Oncorhynchus tshawytscha 0.31 g Hamilton and Buhl 1990b 47000 62,900 97548.09

Rainbow trout 

Oncorhynchus mykiss juvenile Brooke et al. 1985 12000 24,000 82293.77

Oncorhynchus mykiss alevin Buhl and Hamilton 1991 41000 196,460 329848.1

Oncorhynchus mykiss juvenile Buhl and Hamilton 1991 41000 13,501 22667.61

Oncorhynchus mykiss Spehar 1986 12000 47,000 161158.6

Arctic grayling 

Thymallus arcticus alevin Buhl and Hamilton 1991 41000 41,800 70180.45

Thymallus arcticus juvenile Buhl and Hamilton 1991 41000 75,240 126324.8

Fathead minnow

Pimephales promeles Adams 1976 - 11,800 -

Pimephales promeles Adams 1976 - 11,000 -

Pimephales promeles Adams 1976 - 12,500 -

Pimephales promelas juvenile Brooke et al. 1985 12000 2,300 7886.486

Pimephales promelas Spehar 1986 12000 5,500 18858.99

Pimephales promelas GLEC 1998 24000 6,210 14233

Pimephales promelas GLEC 1998 160000 10,800 8218.538

Pimephales promelas GLEC 1998 474000 18,000 7286.649

Pimephales promelas GLEC 1998 906000 42,100 11695.65

Colorado squawfish 

Ptychocheilus lucius fry Hamilton 1995 164000 27,588 20694.67

Ptychocheilus lucius 0.4-1.1 g

juvenile

Hamilton 1995 164000 119,548 89676.92

Ptychocheilus lucius 1.7 g

juvenile

Hamilton 1995 164000 138,358 103786.9

Ptychocheilus lucius larva Buhl and Hamilton 1996 174000 13,580 9842.351

Ptychocheilus lucius juvenile Buhl and Hamilton 1996 174000 42,780 31005.58

Ptychocheilus lucius 0.024-0.047

g

Hamilton and Buhl 1997a 97000 88,000 89571.65
Species Age/ Size Data Source [Sulfate] 

(μg/L)

LC50 or

EC50

(μg/L) 

Adjusted

LC50

A-6 Draft November 12, 2004

Bonytail 

Gila elegans fry Hamilton 1995 164000 22,990 17245.56

Gila elegans 1.1 g

juvenile

Hamilton 1995 164000 102,828 77134.7

Gila elegans 2.6 g

juvenile

Hamilton 1995 164000 90,706 68041.58

Gila elegans larva Buhl and Hamilton 1996 174000 14,570 10559.87

Gila elegans juvenile Buhl and Hamilton 1996 174000 24,010 17401.68

Razorback sucker 

Xyrauchen texanus fry Hamilton 1995 164000 20,064 15050.67

Xyrauchen texanus 0.9 g

juvenile

Hamilton 1995 164000 15,048 11288.00

Xyrauchen texanus 2.0 g

juvenile

Hamilton 1995 164000 10,450 7838.892

Xyrauchen texanus larva Buhl and Hamilton 1996 174000 13,910 10081.52

Xyrauchen texanus juvenile Buhl and Hamilton 1996 174000 7,620 5522.733

Xyrauchen texanus 0.006-0.042

g

Hamilton and Buhl 1997a 97000 15,900 16183.97

Flannelmouth sucker

Catostomus latipinnis 12-13 days Hamilton and Buhl 1997b 97000 26,900 27380.43

Channel catfish

Ictalurus punctatus juvenile Brooke et al. 1985 12000 66,000 226307.9

Bluegill

Lepomis macrochirus juvenile Brooke et al. 1985 12000 63,000 216021.1
B-1 Draft November 12, 2004

APPENDIX B

TOXICITY OF SELENIUM TO AQUATIC PLANTS
B-2 Draft November 12, 2004

Toxicity to Aquatic Plants

Selenite

Data are available on the toxicity of selenite to 13 species of freshwater algae and plants (Table B-1). 

Results ranged from an LC50 of 70,000 μg/L for the green alga, Chlorella ellipsoidea (Shabana and ElAttar 1995) to 522 μg/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 μg/L decreased the

dry weight of Selenastrum capricornutum (Table F-1). Wehr and Brown (1985) reported that 320 μg/L

increased the growth of the alga Chrysochromulina breviturrita. Thus, the sensitivities of freshwater algae

to selenite cover about the same range as the acute and chronic sensitivities of freshwater animals.

The 96-hr EC50 for the saltwater diatom, Skeletonema costatum, is 7,930 μg/L, based on reduction in

chlorophyll a (Table B-1). Growth of Chlorella sp., Platymonas subcordiformis, and Fucus spiralis

increased at selenite concentrations from 2.6 to 10,000 μg/L (Table F-1). 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 μg/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 were affected by concentrations ranging from 100 to 40,000 μg/L

(Table B-1). Blue-green algae appear to be more tolerant to selenate with 1,866 μg/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 μg/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 μg/L

selenate for 14 days (Boisson et al. 1995). Seven other saltwater algal species investigated by Wong and
B-3 Draft November 12, 2004

Oliveira (1991a) exhibited NOEC growth values that ranged from 1,043 to 104,328 μg/L. At 10,000

μg/L, selenate is lethal to four species of saltwater phytoplankton and lower concentrations increase or

decrease growth (Table F-1). Wheeler et al. (1982) reported that concentrations as low as 10 μg/L reduced

growth of Porphyridium cruentum (Table F-1).

Although selenite appears to be more acutely and chronically 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 Anabaena cylindrica, Anabaena flos-aquae, Anabaena variabilis, Anacystis nidulans,

and Scenedesmus dimorphus (Kiffney 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 μg/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.
B-4 Draft November 12, 2004

Table B-1. Toxicity of Selenium to Aquatic Plants

Species Chemical

Hardness

(mg/L as

 CaCO3) 

Duration

 (days) Effect

Concentration

 (μg/L)a Reference

FRESHWATER SPECIES

Selenium (IV)

Green alga,

Chlorella vulgaris

Sodium

selenite

- 90-120 Reduced 

growth

5,480 De Jong 1965

Green alga,

Chlorella

ellipsoidea

Sodium

selenite

- 7 EC50 70,000 Shabana and ElAttar 1995

Green alga,

Scenedesmus

dimorphus

Sodium

selenite

- 14 Reduced 

growth

24,000 Moede et al.

1980

Green alga,

Scenedesmus

quadricauda

Sodium

selenite

- 8 Incipient

inhibition

522 Bringmann and

Kuhn 1977a;

1978a,b; 1979;

1980b

Green alga,

Scenedesmus

quadricauda

Sodium 

selenite

- 8 Incipient

inhibition

2,500 Bringmann and

Kuhn 1959a

Green alga,

Selenastrum

capricornutum

Sodium

selenite

- 4 EC50 2,900 Richter 1982

Green alga,

Selenastrum

capricornutum

Sodium

selenite

- 6 EC50 65,000 Ibrahim and

Spacie 1990

Blue-green alga,

Anabaena

constricta

Sodium

selenite

- 7 EC50 67,000 Shabana and ElAttar 1995

Blue-green alga,

Anabaena

cylindrica

Sodium

selenite

- 14 Reduced 

growth

24,000 Moede et al.

1980

Blue-green alga,

Anabaena flosaquae

Sodium

selenite

- 10 Reduced

chlorophyll a

1,866 Kiffney and

Knight 1990

Blue-green alga,

Anabaena

variabilis

Sodium

selenite

- 6-18 LC50 15,000b Kumar and

Prakash 1971

Blue-green alga,

Anacystis

nidulans

Sodium

selenite

- 10-18 LC50 30,000b Kumar and

Prakash 1971

Blue-green alga,

Microcystis

aeruginisa

Sodium

selenite

- 8 Incipient

inhibition

9,400

 (9,300)

Bringmann and

Kuhn 1976;

1978a,b
Table B-1. Toxicity of Selenium to Aquatic Plants (cont.)

Species Chemical

Hardness

(mg/L as

 CaCO3) 

Duration

 (days) Effect

Concentration

 (μg/L)a Reference

B-5 Draft November 12, 2004

Alga,

Euglena gracilis

- - 15 Reduced 

growth

5,920 Bariaud and

Mestre 1984

Duckweed,

Lemna minor

- - 4 EC50 2,400 Wang 1986

Duckweed,

Lemna minor

Sodium

selenite

- 14 EC50

(mult. rate)

3,500 Jenner and

JanssenMommen 1993

Duckweed,

Lemna minor

Sodium

selenite

- 14 NOEC

(mult. rate)

800 Jenner and

JanssenMommen 1993

Selenium (VI)

Green alga,

Ankistrodesmus

falcatus

Sodium

selenate

- 14 Did not

reduce

growth

10 Vocke et al.

1980

Green alga,

Scenedesmus

dimorphus

Sodium

selenate

- 14 Reduced 

growth

22,100 Moede et al.

1980

Green alga,

Scenedesmus

obliquus

Sodium

selenate

- 14 Reduced 

growth

100 Vocke et al.

1980

Green alga,

Selenastrum

capricornutum

Sodium 

selenate

- 14 Reduced 

growth

300 Vocke et al.

1980

Green alga,

Selenastrum

capricornutum

Sodium

selenate

- 4 EC50 199 Richter 1982

Green alga,

Selenastrum

capricornutum

Sodium

selenate

- 6 EC50 <40,000 Ibrahim and

Spacie 1990

Blue-green alga,

Anabaena

cylindrica

Sodium

 selenate

- 14 Reduced 

growth

22,100 Moede et al.

1980

Blue-green alga,

Anabaena flosaquae

Sodium

selenate

- 10 Reduced

chlorophyll a

1,866 Kiffney and

Knight 1990

Blue-green alga,

Anacystis

nidulans

Sodium 

selenate

- 6-18 EC50 39,000b Kumar and

Prakash 1971
Table B-1. Toxicity of Selenium to Aquatic Plants (cont.)

Species Chemical

Hardness

(mg/L as

 CaCO3) 

Duration

 (days) Effect

Concentration

 (μg/L)a Reference

B-6 Draft November 12, 2004

Blue-green alga,

Anabaena

viriabilis

Sodium

selenate

- 10-18 EC50 17,000b Kumar and

Prakash 1971

Blue-green alga,

Microcoleus

vaginatus

Sodium

selenate

- 14 Reduced 

growth

10,000 Vocke et al.

1980

Duckweed,

Lemna minor

Sodium

selenate

- 14 EC50

(mult. rate)

11,500 Jenner and

JanssenMommen 1993

Duckweed,

Lemna minor

Sodium

selenate

- 14 NOEC

(mult. Rate)

>2,400 Jenner and

JanssenMommen 1993
Table B-1. Toxicity of Selenium to Aquatic Plants (cont.)

B-7 Draft November 12, 2004

Species Chemical

Salinity

 (g/kg) 

Duration

 (days) Effect

Concentration

 (μg/L)a Reference

SALTWATER SPECIES

Selenium (IV)

Green alga,

Dunaliella

tertiolecta

Sodium

selenite

- 60 NOEC growth 1,076 Wong and

Oliveira 1991a

Cyanophyceae alga,

Agemenellum

quadruplicatum

Sodium

selenite

- 60 NOEC growth 10,761 Wong and

Oliveira 1991a

Diatom,

Chaetoceros

vixvisibilis

Sodium

selenite

- 60 NOEC growth 1,076 Wong and

Oliveira 1991a

Diatom,

Skeletonema

costatum

Selenious

acidc

- 4 EC50

(reduction in

chlorophyll a)

7,930 U.S. EPA 1978

Coccolithophore,

Cricosphaera

elongata

Sodium

selenite

- 14 Reduced growth 4,570 Boisson et al.

1995

Dinoflagellate,

Amphidinium

carterae

Sodium

selenite

- 60 NOEC growth 10,761 Wong and

Oliveira 1991a

Dinoflagellate,

Peridinopsis borgei

Selenium 

oxide

- 70-75 Maximum 

growth

0.01-0.05 Lindstrom 1985

Eustigmatophyceae

alga,

Nannochloropsis

oculata

Sodium

selenite

- 60 NOEC growth 107,606 Wong and

Oliveira 1991a

Pyrmnesiophyceae

alga,

Isochrysis galbana

Sodium

selenite

- 60 NOEC growth 1,076 Wong and

Oliveira 1991a

Pyrmnesiophyceae

alga,

Pavlova lutheri

Sodiun

selenite

- 60 NOEC growth 1,076 Wong and

Oliveira 1991a
Table B-1. Toxicity of Selenium to Aquatic Plants (cont.)

Species Chemical

Salinity

 (g/kg) 

Duration

 (days) Effect

Concentration

 (μg/L)a Reference

B-8 Draft November 12, 2004

Selenium (VI)

Green alga,

Dunaliella

tertiolecta

Sodium

selenate

- 60 NOEC growth 104,328 Wong and

Oliveira 1991a

Cyanophyceae alga,

Agemenellum

quadruplicatum

Sodium

selenate

- 60 NOEC growth 10,433 Wong and

Oliveira 1991a

Diatom,

Chaetoceros

vixvisibilis

Sodium

selenate

- 60 NOEC growth 1,043 Wong and

Oliveira 1991a

Coccolithophore,

Cricosphaera

elongata

Sodium

selenate

- 14 Reduced growth 41,800 Boisson et al.

1995

Dinoflagellate,

Amphidinium

carterae

Sodium

selenate

- 60 NOEC growth 10,433 Wong and

Oliveira 1991a

Eustigmatophyceae

alga,

Nannochloropsis

oculata

Sodium

selenate

- 60 NOEC growth 10,433 Wong and

Oliveira 1991a

Pyrmnesiophyceae

alga,

Isochrysis galbana

Sodium

selenate

- 60 NOEC growth 10,433 Wong and

Oliveira 1991a

Pyrmnesiophyceae

alga,

Pavlova lutheri

Sodium

selenate

- 60 NOEC growth 104,328 Wong and

Oliveira 1991a

a

 Concentration of selenium, not the chemical.

b

 Estimated from published graph.

c

 Reported by Barrows et al. (1980) in work performed under the same contract.
C-1 Draft November 12, 2004

APPENDIX C

BIOCONCENTRATION AND BIOACCUMUALTION OF SELENIUM
C-2 Draft November 12, 2004

Bioconcentration and Bioaccumulation

Laboratory-Derived

Bioconcentration factors (BCFs) for selenium(IV) that have been obtained with freshwater species range

from a low of 2 for the muscle of rainbow trout to 470 for the bluegill sunfish (Table C-1). Adams (1976)

studied both uptake and elimination of selenium 75 by fathead minnows exposed to mean concentration of

12, 24, and 50 μg/L in the water. He found that concentrations in whole fish and in individual tissues

increased at a rapid rate during the first 8 days and then at a slower rate for the next 88 days. Steady-state

was approached, but not reached, after 96 days. The highest concentrations were found in viscera. 

Elimination of selenium was curvilinear and became asymptotic with the time axis after 96 days. 

Elimination was most rapid from the viscera with a half-life of 5.1 days, but the half-life of selenium in

other tissues was greater than 50 days.

Adams (1976) also conducted uptake studies with rainbow trout exposed for 48 days to selenium(IV) at

water concentrations ranging from 310 to 950 μg/L. Some of the trout died, and concentrations were

somewhat higher in dead fish than in survivors. As with the fathead minnow, the viscera contained more

selenium than gill or muscle. Based on his tests with the two fish species, Adams (1976) concluded that

there was an inverse relationship between BCF and the concentration of selenium(IV) in water.

Gissel-Nielsen and Gissel-Nielsen (1978) exposed juvenile rainbow trout ( Oncorhynchus mykiss) to

waterborne selenium(IV) over a four week period. Exposure to selenium at 100 μg/L raised the selenium

concentration in fish to 2.3 ± 0.02 μg/g dw, without increasing mortality, and steady-state conditions were

shown to have been achieved.

Hodson et al. (1980) exposed rainbow trout to selenium(IV) from fertilization until 44 weeks post-hatch. 

At 53 μg/L selenium in the water, the BCF ranged from 8 L/kg for whole-body to 240 L/kg for liver. They

concluded that selenium in tissues did not increase in proportion to selenium(IV) in water.

Hunn et al. (1987) exposed rainbow trout in a flow-through system to waterborne selenium(IV) for 90

days. The selenium concentration in the water where significant effects were not observed was 21 μg/L 

and the corresponding whole-body tissue level was 0.64 μg/g dw, the data yielding a BCF value of 30.5

L/kg.
C-3 Draft November 12, 2004

Barrows et al. (1980) exposed bluegills to selenious acid for 28 days. They reported a maximum BCF in

the whole fish of 20 L/kg and a half-life of between 1 and 7 days. If bluegills bioconcentrate selenium in

the same manner as the rainbow trout used by Adams (1976), the 28-day exposure might not have been

long enough to reach steady-state.

Lemly (1982) exposed bluegills and largemouth bass to 10 μg of selenium/L for 120 days to determine the

effect of hardness and temperature on uptake and elimination. For bluegills, the geometric mean wholebody BCF at 20° and 30°C was 452 L/kg. For largemouth bass in similar tests, the BCF was 295 L/kg. 

For both species, the spleen, liver, kidney, and heart had higher concentrations than the whole-body. 

Neither water temperature nor hardness had a significant effect on the amount of selenium accumulated in

the tissues after 90 days, although earlier values were influenced. After 30 days in clean water, selenium

concentrations remained unchanged in spleen, liver, kidney, and white muscle, but the half-life for selenium

in gills and erythrocytes was less than 15 days. 

Besser et al. (1993) measured the aqueous bioaccumulation of both waterborne selenium(IV) and

selenium(VI) by bluegill over a 30-day period. Selenium concentrations were monitored radiometrically

with 75 Se- labeled compounds. Bluegills concentrated selenium about equally from both inorganic species

and demonstrated similar aqueous selenium uptake rate constants (about 3 per day at 10 μg of selenium/L). 

A kinetic uptake-depuration model was used to estimate BCFs. Estimated BCFs for both selenium(IV)

and selenium(VI) derived from the data were 56 L/kg.

Bertram and Brooks (1986) exposed adult fathead minnows to sodium selenate in water, in food, and in

food and water together. The food was specially prepared by raising algae in a medium containing

selenium(VI), feeding the algae to daphnids, mixing the exposed daphnids with unexposed daphnids,

dewatering to form a "cake", and freezing for storage. Uptake of selenium(VI) from water (without the

additional selenium in food) reached steady-state within 28 days. The whole-body BCFs ranged from 21 to

52 L/kg and decreased as the concentration in water increased (Table C-1). Uptake of selenium(VI) from

food alone or from food and water together did not reach steady-state in 8 and 11 weeks, respectively. The

uptake of selenium from food and water were additive.

Besser et al. (1993) also determined BCF values for algae and Daphnia magna exposed separately to

waterborne selenium(IV) and selenium(VI). At 10 μg of selenium/L, the BCFs calculated for algae were
C-4 Draft November 12, 2004

1440 L/kg for selenium(IV) and 428 L/kg for selenium(VI). In these laboratory simulated food web studies

(waterborne selenium to algae; algae to Daphnia; and Daphnia to bluegills) concentration factors (CFs) for

the transfer of selenium from algae to Daphnia and Daphnia to bluegill (0.61 and 0.51 L/kg, respectively)

were also determined (Table C-2). Using the BCF and CF data, one can calculate an estimated BAF for

bluegill for this laboratory food chain. An estimated BAF value of 550 L/kg was calculated for a

waterborne exposure of 10 μg/L of 1:1 selenite:selenate to the algae- Daphnia - bluegill web.

A three-trophic level food chain experiment consisting of the alga, Chlorella vulgaris, the rotifer,

Brachionus calyciflorus, and the fathead minnow, Pimephales promelas was conducted by Dobbs et al.

(1996). The three species were exposed to selenium(VI) for 25 days in a three-trophic level system

whereby the organisms were linked in a continuous flow-through system in separate vessels, with each

organism feeding on the trophic level below it. These organisms were continuously exposed for 25 days to

either 0, 110.3, 207.7 or 396.1 μg of total recoverable selenium/L from selenium(VI) in natural creek water

supplemented with nutrients to sustain algal growth. Algal population growth, rotifer standing crop, and

fathead minnow growth were reduced at 207.7, 110.3 and 110.3 μg/L, respectively, after the 25-day

exposure. Bioconcentration factors were found to be dependent on the species, treatment level and length of

exposure, and they ranged between 100 and 1,000 L/kg.

Hamilton et al. (2000) exposed, separately, swim-up larvae of razorback sucker ( Xyrauchen texanus) and

bonytail ( Gila elegans) to waterborne selenium in a simulated Green River, Utah water formulation. The

selenium was 6:1 selenate:selenite, and the measured ambient or base level was 76 μg/L in the razorback

exposure and 73 μg/L in the bonytail exposure. A flow-through system was utilized, and a 90-day partial

life-cycle chronic toxicity study monitoring growth, behavior and mortality was conducted. No chronic

effects were observed in tests conducted at base level. Higher than ambient concentrations were studied

also, but were not selected for use in the BCF derivation due to either observed chronic effects or

abnormally high concentrations of selenium and other metals in the test waters. At 90 days, the wholebody tissue levels of selenium were 3.2 μg/g dw in the razorback and 2.2 μg/g dw in the boneytail,

reflecting BCF values of 42 and 30 L/kg, respectively.
C-5 Draft November 12, 2004

Field-Derived 

Hermanutz et al. (1996) exposed bluegills to selenium(IV) over 221 days in outdoor experimental streams

at Monticello, MN which contained a natural food web. At the end of the 221 days in waters maintained at

a nominal selenium concentration of 2.5 μg/L, the average whole-body fish tissue level of selenium was

4.825 μg/g Se dw (based on a factor of 0.8 moisture content in fish tissue). The resulting BAF value was

1,930 L/kg.

Garcia-Hernandez et al. (2000) collected fish samples form October 1996 to March 1997 in a Sonora,

Mexico wetland. Dissolved selenium concentrations in the water ranged from 5 to 19 μg/l (median of 11

μg/l). Median whole-body concentration of selenium was measured in Tilapia (3.0 μg/g dw), carp ( 3.3

μg/g dw), and largemouth bass ( 5.1 μg/g dw). Resulting BAF values were 273, 300, and 464 L/kg,

respectively.

Kennedy et al. (2000) collected spawning age (3-6 years) cutthroat trout from the Fording River, British

Columbia in 1998. The waters of the river had an average selenium level of 13.9 μg/L at the time of

collection. The tissue (muscle) of the trout contained 12.5 ± 7.7 μg of selenium/g dw. Utilization of these

values provides a field derived muscle BAF of 899 L/kg.

Mason et al. (2000) collected biota in two streams (Blacklick Run and Herrington Creek) in western

Maryland in October 1997, April 1998, and July 1998. Water samples were collected for analysis monthly

over the duration of the study. Numerous fish species, among other organisms, were collected during each

of the sampling periods, and whole-body tissue levels of selenium were measured. In Herrington Creek, the

average water concentration of selenium was found to be 0.33 μg/L, and the average tissue levels of

selenium in the fish were: bullhead (1.35 μg/g dw); sucker (1.55 μg/g dw), trout (1.94 μg/g Se dw), and

chub (1.50 μg/g Se dw). The resulting calculated BAF values were 4,091, 4,697, 5,879, and 4,545 L/kg,

respectively. In Blacklick Creek the average water concentration was 0.39 μg/L, and the average tissue

levels of selenium in fish were: dace (1.79 μg/g dw), trout (1.94 μg/g dw), and sculpin (2.55 μg/g dw). 

Resulting BAF values were 4,590, 4,974, and 6,538 L/kg, respectively. Dry weight values were obtained

from the published wet weight data employing a 0.8 factor for fish moisture content.
C-6 Draft November 12, 2004

Table C-1. Bioconcentration and Bioaccumulation of selenium by fish.

Fish Species

Selenium

Species

Concentration

in Water

 (μg/L) a 

Duration

 (days) 

Tissue

(Concentration )

BCFb

(L/kg)

 BAFb

(L/kg) Reference

LABORATORY DERIVED

Rainbow trout,

Oncorhynchus

mykiss

Sodium

selenite

- 48 Muscle 2 Adams 1976

Rainbow trout,

Oncorhynchus

mykiss

Sodium

selenite

- 48 Whole-body 10c Adams 1976

Rainbow trout,

Oncorhynchus

mykiss

Sodium

selenite

100 28 Whole-body 

(2.3 μg/g )

23 Gissel-Nielsen

and GisselNielsen 1978

Rainbow trout

(embryo),

Oncorhynchus

mykiss

Sodium

selenite

- 308

(posthatch)

Whole-body

(estimate)

42 Hodson

et al. 1980

Rainbow trout

Oncorhorynchus

mykiss

Sodium

selenite

21 90 Whole-body 

(0.64 μg/g)

30.5 Hunn 

et al. 1987

Fathead minnow,

Pimephales

promelas

Sodium

selenite

- 96 Muscle 11.6 Adams 1976

Fathead minnow,

Pimephales

promelas

Sodium

selenite

- 96 Whole-body 17.6 Adams 1976

Fathead minnow

(6-9 mo.),

Pimephales

promelas

Sodium

selenate

10.7 56 Whole-body 52d Bertram and

Brooks 1986

Fathead minnow

(6-9 mo.),

Pimephales

promelas

Sodium

selenate

21.5 56 Whole-body 26d Bertram and

Brooks 1986

Fathead minnow

(6-9 mo.),

Pimephales

promelas

Sodium

selenate

43.5 56 Whole-body 21d Bertram and

Brooks 1986

Bluegill,

Lepomis

macrochirus

Selenious

acid

- 28 Whole-body 20 Barrows 

et al. 1980
Table C-1 continued.

Fish Species

Selenium

Species

Concentration

in Water

 (μg/L) a

 

Duration

 (days) 

Tissue

(Concentration )

BCFb

(L/kg)

 BAFb

(L/kg) Reference

C-7 Draft November 12, 2004

Bluegill,

Lepomis

macrochirus

Sodium

selenite

10

10

10

10

120

120

120

120

Whole-body

Whole-body

Whole-body

Whole-body

450

470

430

460

Lemly 1982

Bluegill,

Lepomis

macrochirus

Selenate 10 30 Whole-body 56 Besser 

et al. 1993

Largemouth bass,

Micropterus

salmoides

Sodium

selenite

10

10

10

10

120

120

120

120

Whole-body

Whole-body

Whole-body

Whole-body

310

300

300

270

Lemly 1982

Bluegill, 

Lepomis

macrochirus

Selenite 10 30 Whole-body 56 Besser 

et al. 1993

Bluegill,

Lepomis

macrochirus

selenite:

selenate 1:1

10 30 Whole-body 550e Besser 

et al. 1993

Razorback suker,

Xyrauchen

texanus

selenate/

selenite f

76 90 Whole-body

(3.2 μg/g)

42 Hamilton 

et al. 2000

Bonytail,

Gila elegans

selenate/

selenite f

73 90 Whole-body

(2.2 μg/g)

30 Hamilton 

et al. 2000

FIELD DERIVED

Bluegill

Lepomis

macrochirus

Selenite 2.5 221 Whole-body

(4.825 μg/g)

1,930 Hermanutz 

et al. 1996

Tilapia sp. Natural f 11 Field Whole-body

(3.0 μg/g)

273 GarciaHernandez 

et al. 2000

Carp,

Cyprinus carpio

Natural f 11 Field Whole-body 

( 3.3 μg/g)

300 GarciaHernandez 

et al. 2000

Largemouth bass,

Micropterus

salmoides

Natural

 f 11 Field Whole-body

(5.1 μg/g)

464 GarciaHernandez 

et al. 2000

Cutthroat trout,

Oncorhynchus

clarki

Natural f 13.9 Field Muscle 

(12.5 μg/g)

899 Kennedy 

et al. 2000

Brown bullhead,

Ictalurus

nebulosus

Natural f

(Herrington

Creek, MD)

0.33 N/Ag

 (10

month 

study)

Whole-body

(1.35 μg/g)

4,091 Mason et al.

2000
Table C-1 continued.

Fish Species

Selenium

Species

Concentration

in Water

 (μg/L) a

 

Duration

 (days) 

Tissue

(Concentration )

BCFb

(L/kg)

 BAFb

(L/kg) Reference

C-8 Draft November 12, 2004

White sucker,

Catostomus

commersoni 

Natural f

(Herrington

Creek, MD)

0.33 N/A (10

month

study)

Whole-body

(1.55 μg/g)

4,697 Mason et al. 

2000

Brook Trout,

Salvelinus

fonticnalis

Natural f

(Herrington

Creek, MD)

0.33 N/A (10

months

study)

Whole-body

(1.94 μg/g)

5,879 Mason et al. 

2000

Creek Chub,

Semotilus

arromaculatus

Natural f

(Herrington

Creek, MD)

0.33 N/A (10

months

study)

Whole-body

(1.50 μg/g)

4,545 Mason et al. 

2000

Mottled Sculpin, 

Cottus bairdi

Natural f

(Blacklick

Run, MD)

0.39 N/A (10

months

study)

Whole-body

(2.55 μg/g)

6,538 Mason et al. 

2000

Blacknose Dace

Rhinchthus

atratulus

Natural f

(Blacklick

Run, MD)

0.39 N/A (10

months

study)

Whole-body

(1.79 μg/g)

4,590 Mason et al. 

2000

Brook Trout

Salvelins

fortinalus

Natural f

(Blacklick

Run, MD)

0.39 N/A (10

months

study)

Whole-body

(1.94 μg/g)

4,974 Mason et al. 

2000

a Measured concentration of selenium.

b Bioconcentration factors (BCFs) and bioaccumulation factors (BAFs) are based on measured concentrations of selenium in

water and in tissue (dry weight).

c Estimated from graph.

d Calculated by dividing the reported equilibrium concentration in tissue (steady-state body burden) by the average measured

concentration in water.

e Laboratory food chain: water–> algae –> daphnia –> bluegill.

f Not speciated.

g N/A not applicable.
C-9 Draft November 12, 2004

Table C-2. Bioconcentration and Bioaccumulation of selenium by other aquatic organisms.

Other Species

Selenium

Form

Concentration

in Water

 (μg/L) a 

Duration

 (days) 

Tissue

(Concentration )

BCFb

(L/kg)

 BAFb

(L/kg) Reference

LABORATORY DERIVED

Algae, 

Chlamydomonas

reinhardtii

Selenite

Selenate 

10

10

4

4

1440 

428

Besser et

al. 1993

Cladoceran,

Daphnia magna

Selenate:

Selenite 

1:1

Selenate:

Selenite 

1:1

156

348

21

21

Whole-body 

(14.7 μg/g)

Whole-body 

(31.7 μg/g)

94 

91

Ingersoll et

al. 1990

Cladoceran,

Daphnia magna

Selenite

Selenate 

10

10

4

4

570e

293e

Besser et

at. 1993

FIELD DERIVED

Ephemeroptera 

Heptageniidae

Selenite

Selenite

2.5

10

221

221

Whole-body

(5.05 μg/g)

Whole-body

(17.30 μg/g)

1,957

1,787

Hermanutz

et al. 1996

Ephemeroptera 

Heptageniidae

Natural f

(Herrington

Creek, MD)

Natural f

(Blacklick

Run, MD)

0.33

0.39

N/A

 (10 mo.

study)

N/A 

(10 mo.

study)

Whole-body

(5.05 μg/g)

Whole-body 

(5.8 μg/g) 

17,600

14,900

Mason et

al. 2000

Chironomidae Natural f

Natural f

14.5

1.58

N/A

(3 yr.

study)

N/A

(3 yr.

study)

Wholebody

(24.7 μg/g) 

Wholebody

(10.4 μg/g) 

1703

6582

Zhang and

Moore

1996

Chironomidae Selenite

Selenite

2.5

10

221

221

Wholebody

(3.61 μg/g) 

Wholebody

(13.60 μg/g)

1399

1405

Hermanutz

et al. 1996

Hydropsychidae Natural

(ite/ate 9:1)

32 N/A Wholebody

(3.1 μg/g)

969 Reash et

al. 1999
Table C-2 Continued.

Other Species

Selenium

Form

Concentration

in Water

 (μg/L) a 

Duration

 (days) 

Tissue

(Concentration )

BCFb

(L/kg)

 BAFb

(L/kg) Reference

C-10 Draft November 12, 2004

Hydropsychidae Natural f

(Herrington

Creek, MD)

Natural f

(Blacklick

Run, MD)

0.33

0.39

N/A

 (10 mo.

study)

N/A 

(10 mo.

study)

Wholebody

(10.5 μg/g)

Wholebody 

(4.6 μg/g)

31,800

11,800

Mason et

al. 2000

Astacidae Natural f

(Herrington

Creek, MD)

Natural f

(Blacklick

Run, MD)

0.33

0.39

N/A

 (10 mo.

study)

N/A 

(10 mo.

study)

Wholebody 

(1.275 μg/g)

Wholebody 

(0.405 μg/g)

3864

1038

Mason et

al. 2000

Periphyton Natural f

(Herrington

Creek, MD)

Natural f

(Blacklick

Run, MD)

0.33

0.39

N/A

 (10 mo.

study)

N/A 

(10 mo.

study)

Whole 

(2.860 μg/g)

Whole 

(0.245 μg/g)

8667

628

Mason et

al. 2000

Bryophytes Natural f

(Herrington

Creek, MD)

Natural f

(Blacklick

Run, MD)

0.33

0.39

N/A

 (10 mo.

study)

N/A 

(10 mo.

study)

Whole 

(1.860 μg/g)

Whole 

(0.780 μg/g)

5636

2000

Mason et

al. 2000

a Measured concentration of selenium.

b Bioconcentration factors (BCFs) and bioaccumulation factors (BAFs) are based on measured concentrations of selenium in

water and in tissue (dry weight).

c Estimated from graph.

d Calculated by dividing the reported equilibrium concentration in tissue (steady-state body burden) by the average measured

concentration in water.

e Laboratory food chain: water–> algae –> daphnia –> bluegill.

f Not speciated.

g N/A not applicable.
D-1 Draft November 12, 2004

APPENDIX D

ENVIRONMENTAL FACTORS AFFECTING 

SELENIUM TOXICITY AND BIOACCUMULATION
D-2 Draft November 12, 2004

Environmental Factors Affecting Selenium Toxicity and Bioaccumulation

A variety of environmental factors have been shown to influence the toxicity/bioaccumulation of selenium. 

A brief summary of the influence of sulfate, hardness, heavy metals, pH, temperature and day length on

selenium toxicity/bioaccumulation is presented below.

Sulfate

In acute toxicity tests and uptake experiments with selenium, sulfate has been shown to antagonize Se

toxicity and Se uptake in plants and animals, frequently with a major effect on Se action. Where multiple

Se forms are used in joint action experiments, Se(VI) is antagonized most by SO4 with Se(IV) and Se(II)

affected to a lesser extent. Sulfate has reduced Se mortality responses by 90 percent and Se uptake to 10

percent of controls or less. Thus, sulfate is a major co-factor in a number of Se toxicity and Se uptake

experiments.

In four acute toxicity tests, sulfate antagonized selenate toxicity in three algae species and the cladoceran

Daphnia. The LC50 values of two desmids (Cosmerium spp.) exposed to selenate plus sulfate were 4x and

8x the LC50 values of selenate only (Sarma and Jayaraman 1984). The growth of Selenastrum

capricornutum increased by 50 percent when 11 or 107 μg/L Se(VI) were combined with 3.3or 33 μg/L

sulfate (Williams et al. 1994). The toxicity of 490 μg/L selenate to D. magna was reduced by 90 percent

mortality by combining it with either 10 or 308 mg/L sulfate. Uptake studies, with one exception,

document sulfate as antagonistic to uptake of selenium. In many cases, Se uptake rates are reduced to 40

to 50 percent of controls (Se alone or lowest SO4 concentration), but there are examples of sulfate reducing

uptake to 20 percent of controls. These examples include a rooted plant (six percent of control rate), an

alga (7 percent), Daphnia (20 percent) and a midge (20 percent). 

Of the two algal species investigated, Chlamydomonas reinhardtii responded less to sulfate and Se(VI) coexposure (Williams et al. 1994) than Selenastrum capricornutum (7 percent low SO4

 rate) (Riedel and

Sanders 1996). Widgeon grass (Ruppia moritima) uptake reductions (Se uptake, high or low sulfate)

occurred most for Se(VI) (6 percent), then Se(IV) (44 percent) and Se(II) (56 percent) (Bailey et al. 1995).
D-3 Draft November 12, 2004

Experiments with Daphnia show no interaction of Se and SO4 in a microcosm experiment (Besser et al.

1989). However, other experiments with Se and SO4 show a 43 percent reduction of Se uptake by sulfate

(Hansen et al.1993) and uptake reductions ranging from 20 to 65 percent among three Se(VI) exposures

and two sulfate levels (Ogle and Knight 1996). Se uptake by a midge, Chironomus decorus, was reduced

to 20 to 65 percent of controls in a 48 hour exposure to 6 mg/L Se(VI) and 3 levels of SO4 (Hansen et al.

1993).

Hardness

Acute toxicity tests of selenium forms with hardness as a variable were conducted with an invertebrate and

three fish species. In all cases, water hardness variations did not cause major changes in the acute toxicity

of selenium. LC50 value differences due to hardness were no less than half or more than double the LC50 of

the standard of comparison.

D. magna were exposed to three forms of selenium and one Se mixture in acute toxicity tests (48h LC50) to

determine the effect of soft (46 mg/L CaCO3) and hard (134 mg/L CaCO3) water on selenium toxicity. 

Water hardness did not affect the toxicity of Se(VI) and Se(II), but Se(IV) was slightly more toxic in hard

than soft water (LC50, hard/soft = 0.5), as was the 1:1 mixture of Se(IV) and Se(VI) (LC50, hard/soft = 0.6)

(Ingersoll et al. 1990). Mytilus edulis were exposed to selenite in sea water with salinities of 15, 20, 27

and 30‰ (27‰ was close to the mussel’s natural habitat). Se(IV) influx measured during 2 hours of

exposure demonstrated an effect on uptake as follows: maximum influx at 20‰ ; greatest influx difference

= 0.7 max (34‰ ) (Wang et al. 1996a).

Fry of chinook salmon and coho salmon were exposed for 96 hr to selenate, selenite and a 1:1 mixture in

soft (42 mg/L CaCO3) and hard (211 mg/L CaCO3) water. Advanced fry of chinook salmon were exposed

to Se(II) in brackish water (333 mg/L CaCO3 ). In all cases, variable hardness had no effect on the toxicity

of three forms of selenium or the mixture (Hamilton and Buhl 1990b). 

Young striped bass (Morone saxatilis) exhibited some differential susceptibility to selenite in hard (285

mg/L CaCO3) vs. soft (40 mg/L CaCO3) water (LC50 hard/soft = 1.8) with Se(IV) in soft water being more

toxic. The LC50 of Se(IV) in 1‰ saline (455 mg/L CaCO3) was not significantly different than Se in soft or

hard water (Palawski et al. 1985).
D-4 Draft November 12, 2004

The Se BCF values for young salmon (Onchorhynchus tsawytscha) exposed for 90 days to a Se(VI):Se(IV)

mixture (6:1) were no different in fresh water (371 mg/L CaCO3) or well water (612 mg/L CaCO3). 

Exposure for 60 days to Se in 1‰ saline reduced the Se BCF to approximately 50% of BCFs for well

water and fresh water (Hamilton and Wiedmeyer 1990).

Heavy Metals

Joint action studies with selenium and metals were conducted with cadmium and mercury, which have been

investigated frequently in this regard, and arsenic and molybdenum. The latter two chemicals were

investigated in a chronic test (at least 3 broods) with Ceriodaphnia dubia at exposure concentrations of the

three chemicals that alone caused chronic mortality and reproductive effects. As+5 or Mo+6, combined with

Se(VI) in a chronic test, reduced reproduction and increased cumulative mortality (Naddy et al. 1995).

Mercury uptake experiments with selenite had opposite results in two separate studies with the marine

mussel, Mytilus edulis. In 30 - 50-day tests, Se(IV) uptake was doubled (Se alone = 0.8 ng/g/d) by joint

exposure to Se (30 μg/L) and HgCl2 (5 μg/L) (Pelletier 1986a). Uptake of Se in a 96-hr study (Se(IV), 2

μg/L; HgCl2, 0.1- 1.0 mg/L) decreased as a function of Hg concentration (r2 = - 0.93) (Micallef and Tyler

1987).

The toxicity of Se(IV) and Se(VI) to a fresh water snail (Lymnaea) was reduced by 55 to 66 percent

mortality by 0.1 mg/L cadmium in an 11-day water exposure. Using growth to evaluate toxicity of

selenium-cadmium pairs in two species of marine phytoplankton (Crypthecodinium sp., Procentrum sp.),

Prevot and Sayer-Gobillard (1986) demonstrated in both species that the toxicity of the higher Se doses

was reduced by cadmium. Cadmium slightly elevated Se(IV) uptake in gill tissues of Carcinas maenas

(marine shore crabs) but Se levels in two other tissues and carapace were no different than Se exposure in a

29-day experiment.

In summary, cadmium mortality effects were consistent in antagonizing the toxicity of selenium, although

the level of antagonism was low to moderate in these two cases. Mercury effects on Se uptake by Mytilus

were not in agreement, i.e. in a 96-hour study, selenium uptake decreased as mercury increased, but in 30 -

50-day tests, mercury enhanced selenium uptake. Both metals are generally toxic which complicates Se-
D-5 Draft November 12, 2004

metal investigations. For example, Se interaction with arsenic or molybdenum were conducted with metal

concentrations that were toxic.

pH, Temperature and Day Length

Except for Se(IV) at acidic pH, pH changes in the range associated with natural waters do not have an

appreciable effect on uptake of selenium. Temperature is a major modifying influence on the interaction of

chemicals and aquatic organisms as shown by sediment storage and Paramecium experiments. Interaction

by low temperature and day length dramatically enhanced the toxicity of Se in fish chronically exposed

under laboratory conditions.

As presented in the chronic section, Lemly (1993b) investigated the effect of temperature and day-length

effects with selenium on juvenile bluegills exposed for 180 days. Selenium exposures included 4.8 μg/L in

water (SeVI:SeIV = 1:1) and Se(II) in food (5.1 μg/g) and simulation of summer conditions and winter

conditions. Functions monitored during the study were percent lipid content of fish (energy reserve),

cumulative mortality, body condition factor, QO2 and gill pathology and blood abnormalities. All of these

major functions were significantly affected by winter simulation plus selenium in experiments designed to

chronically expose bluegills to a combination of selenium and environmental factors that would reflect

actual exposure of natural fish populations to selenium during seasonal change.
E-1 Draft November 12, 2004

APPENDIX E

SITE-SPECIFIC CONSIDERATIONS
E-2 Draft November 12, 2004

Site-specific Considerations

Aquatic organism uptake of selenium by both water column exposure and dietary pathways has prompted a

number of researchers to investigate the toxicity of selenium under site-specific conditions. Previous sitespecific studies have addressed the water-based chronic criterion of 5 μg/L through examination of

environmental variables that could potentially influence the availability and/or accumulation of selenium

within the aquatic ecosystem under consideration, thereby either increasing or decreasing the toxic impact

of selenium on the aquatic community (Adams et al. 1998; Canton and VanDerveer 1997; VanDerveer and

Canton 1997).

Now that the recommended chronic criterion is tissue-based, site-specific factors that affect the

bioaccumulation of selenium are not relevant in the modification of the criterion. Recent studies on the

effects of selenium on bluegill in streams receiving wastewater from a coal ash effluent suggest fish

exposed to Se-laden effluents may exhibit tolerance (Lohner et al. 2001a,b,c). The authors found the

bluegill population receiving the coal ash effluent to have an age class structure and condition indices

similar to reference locations despite having selenium concentration in the ovary and whole-body tissues

twice the level of the FCV. Hematological and biochemical assays using samples from exposed bluegill

have shown a reduced response relative to reference fish,but the authors contend that they are not always

related to selenium. The authors hypothesize that selenium speciation, metabolism, bioavailability and

antagonism are possible reasons for the decreased sensitivity of the resident bluegill population in the ash

stream. To date, no experiments on the success on embryo-larval development have been conducted.

In an effort to determine if a proposed multiple-use water development project (Animas La Plata) would

adversely affect aquatic biota in Colorado and New Mexico, Lemly (1997c) conducted a hazard assessment

of selenium using the Protocol Method (Lemly 1995). Using existing environmental monitoring data, the

hazard assessment indicated that selenium poses a siginificant toxic threat to aquatic biota in the Animas

La Plata Project. Incorporating this information into the proposed water development will substantially

reduce the chances of experiencing significant environmental problems similar to those encountered at

Belews Lake and Kesterson National Wildlife Refuge. Once an aquatic system is impacted with selenium,

it could take several to many years before the biological health of the system can be returned to the original

condition prior to perturbation. The Grassland Water District in central California is an example of an
E-3 Draft November 12, 2004

aquatic system that was contaminated with selenium as a result of subsurface agricultural drainwater used

for wetland management since 1954 (Paveglio et al. 1997). Selenium contamination of aquatic bird food

chains prompted the California State Water Resources Board to mandate the Grassland Water District to

reduce selenium concentration starting in 1985 by essentially filling the wetlands with freshwater only. 

Selenium concentrations in a number of aquatic birds have gradually declined since 1985 (1985 to 1994),

but selenium concentrations in some wintering birds still were above concentrations associated with

impaired reproduction in laboratory and field studies. The authors estimated under the current management

strategy, an additional 1 to 13 years from 1994 are needed for selected species to reach background

selenium levels in liver. Thus, approximately 10 to 20 years are needed at this site to reduce the elevated

levels of selenium in avian species and restore normal reproductive success.
F-1 Draft November 12, 2004

APPENDIX F

OTHER DATA
F-2 Draft November 12, 2004

Other Data

Selenite

Additional data on the lethal and sublethal effects of selenium on aquatic species are presented in Table F1. 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 F-1). 

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 ? g/L. 

Selenate

Dunbar et al. (1983) exposed fed D. magna to selenate for seven days and obtained an LC50 of 1,870 ? g/L. 

This value is in the range of the 48-hr EC50s in Table F-1.

Watenpaugh and Beitinger (1985a) found that fathead minnows did not avoid 11,200 ? g/L selenate during

30-minute exposures (Table F-1). These authors also reported (1985b) a 24-hr LC50 of 82,000 ? g/L for

the same species and they found (1985c) that the thermal tolerance of the species was reduced by 22,200

? g/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 ? g/L, but when adults were

exposed to 20,000 ? g/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 ? g/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 ? g/L, but

50% of 72-day-old juveniles died after four days at 87,000 ? g/L. Exposure of juvenile fish for up to 65

days to concentrations of selenate between 39 and 1,360 ? g/L caused developmental anomalies and

pathological lesions.
F-3 Draft November 12, 2004

Table F-1. Other Data on Effects of Selenium on Aquatic Organisms

Species Chemical

Hardness

(mg/L as

 CaCO3) Duration Effect Concentrationa Reference 

FRESHWATER SPECIES

Selenium (IV)

Green alga,

Scenedesmus

quadricauda

Sodium 

selenite

- 96 hr Incipient

inhibition (river

water)

2,500 Bringmann and

Kuhn 1959a,b

Green alga,

Selenastrum

capricornutum

Sodium 

selenite

- 72 hr Decreased dry

weight and

chlorophyll a

75 Foe and Knight,

Manuscript

Green alga,

Selenastrum

capricornutum

Sodium 

selenite

- 72 hr BCF = 12-21b 10-100 Foe and Knight,

Manuscript

Green alga,

Selenastrum

capricornutum

Sodium 

selenite

- 72 hr BCF = 11,164c 150 Foe and Knight,

Manuscript

Alga,

Chrysochromulina

breviturrita

Selenious 

acid

- 30 days Increased growth 320 Wehr and Brown

1985

Red alga,

Cyanidium

caldarium

Selenious

 acid

- 20 days Inhibited growth 3,958 Albertano and

Pinto 1986

Red alga,

Cyanidioschyzon

merolae

Selenious

acid

- 20 days Inhibited growth 3,140 Albertano and

Pinto 1986

Red alga,

Galdieria

sulphuraria

Selenious

acid

- 20 days Inhibited growth 790 Albertano and

Pinto 1986

Algae (diatoms),

Mixed population

Sodium 

selenite

- 18 days Inhibited growth 11,000 Patrick et al.

1975

Bacterium,

Escherichia coli

Sodium 

selenite

- - Incipient

inhibition

90,000 Bringmann and

Kuhn 1959a

Bacterium,

Pseudomonus putida

Sodium 

selenite

- 16 hr Incipient

inhibition

11,400

(11,200)

Bringmann and

Kuhn 1976;

1977a; 1979;

1980b

Protozoan,

Entosiphon sulcatum

Sodium 

selenite

- 72 hr Incipient

inhibition

1.8

(1.9)

Bringmann 1978;

Bringmann and

Kuhn 1979;

1980b; 1981

Protozoan,

Microreqma

heterostoma

Sodium 

selenite

- 28 hr Incipient

inhibition

183,000 Bringmann and

Kuhn 1959b
Table F-1. Other Data on Effects of Selenium on Aquatic Organisms (continued)

Species Chemical

Hardness

(mg/L as

 CaCO3) Duration Effect Concentrationa Reference 

F-4 Draft November 12, 2004

Protozoan,

Chilomonas

paramecium

Sodium 

selenite

- 48 hr Incipient

inhibition

62 Bringmann and

Kuhn 1981;

Bringmann et al.

1980

Protozoan,

Uronema parduezi

Sodium 

selenite

- 20 hr Incipient

inhibition

118 Bringmann and

Kuhn 1980a;

1981

Snail,

Lymnaea stagnalis

Sodium 

selenite

- 7.5 days LT50 3,000 Van Puymbroeck

et al. 1982

Cladoceran,

Daphnia magna

Sodium 

selenite

- 48 hr EC50 (river

water)

2,500 Bringmann and

Kuhn 1959a,b

Cladoceran,

Daphnia magna

Sodium 

selenite

214 24 hr LC50 16,000 Bringmann and

Kuhn 1977a

Cladoceran,

Daphnia magna

Sodium 

selenite

214 24 hr EC50

(swimming)

9.9 Bringmann and

Kuhn 1977b

Cladoceran,

Daphnia magna

Sodium 

selenite

329 48 hr

96 hr

14 days

EC50 (fed) 710

430

430

Halter et al. 1980

Cladoceran (<24 hr),

Daphnia magna

Sodium 

selenite

- 48 hr

21 days

EC50 (fed) 685

160

Adams and

Heidolph 1985

Cladoceran 

(5th instar),

Daphnia magna

Sodium 

selenite

- 48 hr LC50 (fed) 680 Johnston 1987

Cladoceran,

Daphnia magna

Selenious 

acid

220d 48 hr LC50 (fed) 1,200 Kimball,

Manuscript

Cladoceran

(preadult),

Daphnia pulex

Sodium 

selenite

42 24 hr Did not reduce

oxygen

consumption or

filtering rate

>498 Reading and

Buikema 1980

Ostracod,

Cyclocypris sp.

Sodium 

selenite

100.8 48 hr LC50 130,000 Owsley 1984

Amphipod,

Hyalella azteca

Sodium 

selenite

329 14 days LC50 (fed) 70 Halter et al. 1980

Amphipod 

(2 mm length),

Hyalella azteca

Sodium 

selenite

133 48 hr LC50 623 Brasher and Ogle

1993

Amphipod 

(2 mm length),

Hyalella azteca

Sodium

selenite

133 10 days LC50

(fed)

312 Brasher and Ogle

1993
Table F-1. Other Data on Effects of Selenium on Aquatic Organisms (continued)

Species Chemical

Hardness

(mg/L as

 CaCO3) Duration Effect Concentrationa Reference 

F-5 Draft November 12, 2004

Amphipod 

(2 mm length),

Hyalella azteca

Sodium

selenite

133 24 days LOEC

reproduction

(static-renewal)

200 Brasher and Ogle

1993

Midge (first instar),

Chironomus riparius

Sodium

selenite

134 48 h LC50 7,950 Ingersoll et al.

1990

Midge (first instar),

Chironomus riparius

Sodium

selenite

40-48 48 h LC50 14,600 Ingersoll et al.

1990

Coho salmon (fry),

Oncorhynchus

kisutch

Sodium 

selenite

325 43 days LC50 160 Adams 1976

Rainbow trout (fry),

Oncorhynchus

mykiss

Sodium 

selenite

334 21 days LC50 460 Adams 1976

Rainbow trout (fry),

Oncorhynchus

mykiss

Sodium 

selenite

334 21 days Reduced growth 250 Adams 1976

Rainbow trout,

Oncorhynchus

mykiss

Sodium 

selenite

330 5 days LC50 2,700

2,750

Adams 1976

Rainbow trout,

Oncorhynchus

mykiss

Sodium 

selenite

325 48 days LC50 500 Adams 1976

Rainbow trout,

Oncorhynchus

mykiss

Sodium 

selenite

325 96 days LC50 280 Adams 1976

Rainbow trout

(juvenile),

Oncorhynchus

mykiss

Sodium 

selenite

- 4 wk MATC 

survival

200 Gissel-Nielsen

and GisselNielsen 1978

Rainbow trout

(juvenile),

Oncorhynchus

mykiss

Sodium 

selenite

- 4 wk MATC 

survival

4.7

μg/g dw 

(whole-body)

Gissel-Nielsen

and GisselNielsen 1978

Rainbow trout

(juvenile),

Oncorhynchus

mykiss

Sodium 

selenite

- 4 wk BCF = 23 100 Gissel-Nielsen

and GisselNielsen 1978

Rainbow trout

(juvenile),

Oncorhynchus

mykiss

Sodium 

selenite

- 42 wk MATC growth

(dietary only

exposure)

>9.96

μg Se/g dw

(food)

Goettl and

Davies 1978
Table F-1. Other Data on Effects of Selenium on Aquatic Organisms (continued)

Species Chemical

Hardness

(mg/L as

 CaCO3) Duration Effect Concentrationa Reference 

F-6 Draft November 12, 2004

Rainbow trout

(juvenile),

Oncorhynchus

mykiss

Sodium 

selenite

- 42 wk MATC survival

(dietary only

exposure)

5.34

μg Se/g dw

(food)

Goettl and

Davies 1978

Rainbow trout,

Oncorhynchus

mykiss

Sodium 

selenite

135 9 days LC50 7,020 Hodson et al.

1980

Rainbow trout,

Oncorhynchus

mykiss

Sodium 

selenite

135 96 hr

9 days

LC50

(fed)

7,200

5,410

Hodson et al.

1980

Rainbow trout,

Oncorhynchus

mykiss

Sodium 

selenite

135 96 hr

9 days

LC50

(fed)

8,200

6,920

Hodson et al.

1980

Rainbow trout,

Oncorhynchus

mykiss

Sodium 

selenite

135 41 days LOAEC

(Reduced hatch

of eyed embryos)

26 Hodson et al.

1980

Rainbow trout,

Oncorhynchus

mykiss

Sodium 

selenite

135 50 wk Decreased iron

in blood and red

cell volume

53 Hodson et al.

1980

Rainbow trout

(fertilized egg),

Oncorhynchus

mykiss

Sodium 

selenite

135 44 wk BCF = 33.2

BCF = 21.1

53 Hodson et al.

1980

Rainbow trout

(embryo),

Oncorhynchus

mykiss

Sodium 

selenite

- 120 hr Did not reduce

survival or time

to hatch

10,000 Klaverkamp et

al. 1983b

Rainbow trout,

Oncorhynchus

mykiss

Sodium

selenite

- 90 days Chronic value for

survival

14 Mayer et al.

1986

Rainbow trout 

(sac fry),

Oncorhynchus

mykiss

Sodium 

selenite

272 90 days LC50 55.2e Hunn et al. 1987

Rainbow trout 

(sac fry),

Oncorhynchus

mykiss

Sodium 

selenite

272 90 days MATC

survival

31.48 Hunn et al. 1987

Rainbow trout 

(egg ),

Oncorhynchus

mykiss

Sodium 

selenite

- 96 hr BCF = 17.5

BCF = 3.5

0.4

45.6

Hodson et al.

1986
Table F-1. Other Data on Effects of Selenium on Aquatic Organisms (continued)

Species Chemical

Hardness

(mg/L as

 CaCO3) Duration Effect Concentrationa Reference 

F-7 Draft November 12, 2004

Rainbow trout

(embryo),

Oncorhynchus

mykiss

Sodium 

selenite

- 96 hr BCF = 3.1

BCF = 3.0

0.4

45.6

Hodson et al.

1986

Rainbow trout 

(sac-fry),

Oncorhynchus

mykiss

Sodium 

selenite

- 96 hr BCF = 13.1

BCF = 1.6

0.4

45.6

Hodson et al.

1986

Rainbow trout

(swim-up fry)

Oncorhynchus

mykiss

Sodium 

selenite

- 96 hr BCF = 80.3

BCF = 20.2

0.4

45.6

Hodson et al.

1986

Northern pike,

Esox lucius

Sodium 

selenite

10.2 76 hr LC50 11,100 Klaverkamp et

al. 1983a

Goldfish,

Carassius auratus

Selenium

dioxide

157 14 days LC50 6,300 Cardwell et al.

1976a,b

Goldfish,

Carassius auratus

Sodium 

selenite

- 10 days Mortality 5,000 Ellis 1937; Ellis

et al. 1937

Goldfish,

Carassius auratus

Sodium 

selenite

- 46 days Gradual anorexia

and mortality

2,000 Ellis et al. 1937

Goldfish,

Carassius auratus

Selenium

dioxide

- 7 days LC50 12,000 Weir and Hine

1970

Goldfish,

Carassius auratus

Selenium

dioxide

- 48 hr Conditional

avoidance

250 Weir and Hine

1970

Fathead minnow,

Pimephales

promelas

Selenium

dioxide

157 9 days LC50 2,100 Cardwell et al.

1976a,b

Fathead minnow,

Pimephales

promelas

Sodium 

selenite

329 96 hr LC50

(fed)

1,000 Halter et al. 1980

Fathead minnow,

Pimephales

promelas

Sodium 

selenite

329 14 days LC50

(fed)

600 Halter et al. 1980

Fathead minnow,

Pimephales

promelas

Selenious 

acid

220d 8 days LC50

(fed)

420 Kimball,

Manuscript

Creek chub,

Semotilus

atromaculatus

Selenium

dioxide

- 48 hr Mortality ?12,000 Kim et al. 1977
Table F-1. Other Data on Effects of Selenium on Aquatic Organisms (continued)

Species Chemical

Hardness

(mg/L as

 CaCO3) Duration Effect Concentrationa Reference 

F-8 Draft November 12, 2004

Bluegill,

Lepomis

macrochirus

Sodium 

selenite

318 48 days LC50 400 Adams 1976

Bluegill,

Lepomis

macrochirus

Selenium

dioxide

157 14 days LC50 12,500 Cardwell et al.

1976a,b

Bluegill (juvenile),

Lepomis

macrochirus

Sodium 

selenite

16 323 days MATC larval

survival

(dietary only

exposure)

19.75

μg Se/g dw 

(food)

Woock et al.

1987

Bluegill (juvenile),

Lepomis

macrochirus

Sodium 

selenite

25

and

200

120 days No mortality >10 Lemly 1982

Largemouth bass

(juvenile),

Micropterus

salmoides

Sodium 

selenite

25

and

200

120 days No mortality 10 Lemly 1982

Yellow perch,

Perca flavescens

Sodium 

selenite

10.2 10 days LC50 4,800 Klaverkamp et

al. 1983a,b

African clawed frog,

Xenopus laevis

Sodium 

selenite

- 7 days LC50 1,520 Browne and

Dumont 1980

African clawed frog,

Xenopus laevis

Sodium 

selenite

- 1-7 days Cellular damage 2,000 Browne and

Dumont 1980

Selenium (VI)

Alga,

Chrysochromulina

breviturrita

- - 30 days Increased 

growth

50 Wehr and Brown

1985

Snail,

Lymnaea stagnalis

Sodium 

selenate

- 6 days LT50 15,000 Van Puymbroeck

et al. 1982

Cladoceran,

Daphnia magna

Sodium 

selenate

129.5 7 days LC50

(fed)

1,870 Dunbar et al.

1983

Cladoceran

(juvenile),

Daphnia magna

Sodium 

selenate

- 48 hr LC50

(fed)

550 Johnston 1987

Cladoceran 

(5th instar),

Daphnia magna

Sodium 

selenate

- 48 hr LC50

(fed)

750 Johnston 1987
Table F-1. Other Data on Effects of Selenium on Aquatic Organisms (continued)

Species Chemical

Hardness

(mg/L as

 CaCO3) Duration Effect Concentrationa Reference 

F-9 Draft November 12, 2004

Cladoceran 

(5th instar),

Daphnia magna

Sodium 

selenate

- 90 hr 42% of organisms had visible

changes in gut

morphology

250 Johnston 1989

Amphipod 

(2 mm length),

Hyalella azteca

Sodium

selenate

133 48 hr LC50 2378 Brasher and Ogle

1993

Amphipod 

(2 mm length),

Hyalella azteca

Sodium 

selenate

133 10 days LC50

(fed)

627 Brasher and Ogle

1993

Amphipod 

(2 mm length),

Hyalella azteca

Sodium 

selenate

133 24 days LOEC

reproduction 

(static renewal)

>700 Brasher and Ogle

1993

Midge (first instar),

Chironomus riparius

Sodium

selenate

134 48 h LC50 16,200 Ingersoll et al.

1990

Midge (first instar),

Chironomus riparius

Sodium

selenate

40-48 48 h LC50 10,500 Ingersoll et al.

1990

Rainbow trout

(embryo, larva),

Oncorhynchus

mykiss

Sodium 

selenate

104

(92-110)

28 days EC50 (death and

deformity)

5,000

(4,180)

(5,170)

Birge 1978;

Birge and Black

1977; Birge et al.

1980

Goldfish

(embryo, larva),

Carrassius auratus

Sodium 

selenate

195 7 days EC50 (death and

deformity)

8,780 Birge 1978

Goldfish,

Carassius auratus

Sodium 

selenate

- 24 hr BCF = 1.42

BCF = 1.15

BCF = 1.47

BCF = 0.88

BCF = 1.54

0.45

0.9

1.35

2.25

4.5

Sharma and

Davis 1980

Fathead minnow,

Pimephales

promelas

Sodium 

selenate

337.9 48 days LC50 2,000 Adams 1976

Fathead minnow,

Pimephales

promelas

Sodium 

selenate

338 48 days LC50 1,100 Adams 1976

Fathead minnow,

Pimephales

promelas

- 51 30 min No avoidance 11,200 Watenpaugh and

Beitinger 1985a

Fathead minnow,

Pimephales

promelas

- - 24 hr LC50 82,000 Watenpaugh and

Beitinger 1985b
Table F-1. Other Data on Effects of Selenium on Aquatic Organisms (continued)

Species Chemical

Hardness

(mg/L as

 CaCO3) Duration Effect Concentrationa Reference 

F-10 Draft November 12, 2004

Fathead minnow,

Pimephales

promelas

- - 24 hr Reduced thermal

tolerance

22,200 Watenpaugh and

Beitinger 1985c

Fathead minnow,

Pimephales

promelas

Sodium 

selenate

44-49 7 days Chronic value -

growth

Chronic valuegrowth

Chronic valuesurvival

1,739

561

2,000

Norberg-King

1989

Fathead minnow,

Pimephales

promelas

Sodium

selenate

160-180 24 hr No effect on

reproductive

behavior

36,000 Pyron and

Beitinger 1989

Fathead minnow,

Pimephales

promelas

Sodium

selenate

160-180 24 hr Edema in larvae

produced from

adults exposed to

Selenium VI

20,000 Pyron and

Beitinger 1989

Channel catfish

(embryo, fry),

Ictalurus punctatus

Sodium 

selenate

90 8.5-9

days

Induced albinism - Westerman and

Birge 1978

Narrow-mouthed

toad

(embryo, larva),

Gastrophryne

carolinensis

Sodium 

selenate

195 7 days EC50 (death and

deformity)

90 Birge 1978;

Birge and Black

1977; Birge et al.

1979a

Organo-selenium

Bluegill (juvenile),

Lepomis

macrochirus

Seleno-Lmethionine16 323 days MATC larval

survival

(dietary only

exposure)

20.83

μg Se/g dw 

(food)

Woock et al.

1987

Bluegill (juvenile),

Lepomis

macrochirus

Seleno-Lmethionine283 90 days EC20 survival

(dietary only

exposure)

>13.4

μg/g dw 

(food)

Cleveland et al.

1993

Bluegill 

(2 yr and adult),

Lepomis

macrochirus

Selenium - field NOEC

deformities

53.83

μg Se/g dw 

(liver)

Reash et al. 1999

Bluegill 

(2 yr and adult),

Lepomis

macrochirus

Selenium - field NOEC

deformities

23.38

μg Se/g dw 

(ovaries)

Reash et al. 1999
Table F-1. Other Data on Effects of Selenium on Aquatic Organisms (continued)

Species Chemical

Hardness

(mg/L as

 CaCO3) Duration Effect Concentrationa Reference 

F-11 Draft November 12, 2004

Redear sunfish

(adult), 

Lepomis

microlophus

Selenium - field LOEC Adverse

histopathological

alterations

<38.15

μg Se/g dw

Sorensen 1988

Selenium Mixtures

Phytoplankton,

Mixed population

Selenium - field Reduced growth

rates

18 Riedel et al.

1991

Cladoceran 

(<24 hr),

Daphnia magna

SeleniteSelenate

mixture

138 21 days MATC

growth

115.2

μg Se/L

Ingersoll et al.

1990

Cladoceran 

(<24 hr),

Daphnia magna

SeleniteSelenate

mixture

138 21 days MATC

productivity

21.59 ? g/g dw

(whole-body)

Ingersoll et al.

1990

Midge (<24-hr),

Chironomus riparius

SeleniteSelenate

mixture

138 30 days MATC 

emergence

503.6 Ingersoll et al.

1990

Bluegill (juvenile),

Lepomis

macrochirus

SeleniteSelenate

mixture

283 60 days NOEC survival 340 Cleveland et al.

1993

Bluegill (juvenile),

Lepomis

macrochirus

SeleniteSelenate

mixture

283 60 days EC20 survival 4.07

μg/g dw 

(whole body)

Cleveland et al.

1993

Species Chemical

Salinity

 (g/kg) Duration Effect

Concentration 

 (ug/L)a Reference

SALTWATER SPECIES

Selenium (IV)

Anaerobic

bacterium,

Methanococcus

vannielli

Sodium 

selenite

- 110 hr Stimulated growth 79.01 Jones and

Stadtman

1977

Bacterium,

Vibrio fisheri

Sodium 

selenite

- 5 min 50% decrease in

light output

(Microtox®)

68,420 Yu et al.

1997

Green alga,

Chlorella sp.

Sodium 

selenite

32 14 days 5-12% increase in

growth

10-10,000 Wheeler et

al. 1982
Table F-1. Other Data on Effects of Selenium on Aquatic Organisms (continued)

Species Chemical

Salinity

 (g/kg) Duration Effect

Concentration 

 (ug/L)a Reference

F-12 Draft November 12, 2004

Green alga,

Platymonas

subcordiformis

Sodium 

selenite

32 14 days 23% increase in

growth

100-10,000 Wheeler et

al. 1982

Green alga,

Dunaliella

primolecta

Sodium 

selenite

32 20 days Increased growth;

induced glutathione

peroxidase

4,600 Gennity et al.

1985a,b

Diatom, 

Skeletonema

costatum

Selenium

dioxide

- 5 days BCF = 18,000

BCF = 16,000

BCF = 10,000

0.06

0.79

3.6

Zhang et al.

1990

Diatom, 

Chaetoceros

muelleri

Selenium

dioxide

- 6 days BCF = 337,000

BCF = 65,000

BCF = 5,000

0.06

0.79

3.6

Zhang et al.

1990

Diatom, 

Phaeodactylum

tricornutum

Selenium

dioxide

- 8 days BCF = 109,000

BCF = 27,000

BCF = 7,000

0.06

0.79

3.6

Zhang et al.

1990

Diatom,

Thallassiosira

aestivalis

Selenium 

oxide

29-30 72 hr No effect on cell

morphology

78.96 Thomas et al.

1980a

Brown alga,

Fucus spiralis

Sodium 

selenite

- 60 days 1355% increase in

growth of thalli

2.605 Fries 1982

Red alga,

Porphyridium

cruentum

Sodium 

selenite

32 27 days Increase growth;

induced glutathione

peroxidase

4,600 Gennity et al.

1985a,b

Selenium (VI)

Bacterium,

Vibrio fisheri

Sodium 

selenate

- 15 min 50% decrease in

light output

(Microtox®)

3,129,288 Yu et al.

1997

Green alga,

Chlorella sp.

Sodium 

selenate

32 14 days No effect on rate of

cell

10-1,000 Wheeler et

al. 1982

Green alga,

Chlorella sp.

Sodium 

selenate

32 4-5 days 100% mortality 10,000 Wheeler et

al. 1982

Green alga,

Dunaliella

primolecta

Sodium 

selenate

32 14 days No effect on rate of

cell population

growth

10-100 Wheeler et

al. 1982

Green alga,

Dunaliella

primolecta

Sodium 

selenate

32 14 days 71% reduction in

rate of cell

population growth

1,000 Wheeler et

al. 1982

Green alga,

Dunaliella

primolecta

Sodium 

selenate

32 4-5 days 100% mortality 10,000 Wheeler et

al. 1982
Table F-1. Other Data on Effects of Selenium on Aquatic Organisms (continued)

Species Chemical

Salinity

 (g/kg) Duration Effect

Concentration 

 (ug/L)a Reference

F-13 Draft November 12, 2004

Green alga,

Platymonas

subcordiformis

Sodium 

selenate

32 14 days No effect on rate of

cell population

growth

10 Wheeler et

al. 1982

Green alga,

Platymonas

subcordiformis

Sodium 

selenate

32 14 days 16% decrease in

rate of cell

population growth

100 Wheeler et

al. 1982

Green alga,

Platymonas

subcordiformis

Sodium 

selenate

32 14 days 50% decrease in

rate of cell

population growth

1,000 Wheeler et

al. 1982

Green alga,

Platymonas

subcordiformis

Sodium 

selenate

32 4-5 days 100% mortality 10,000 Wheeler et

al. 1982

Brown alga,

Fucus spiralis

Sodium 

selenate

- 60 days 160% increase in

growth rate of thalli

2.605 Fries 1982

Red alga,

Porphridium

cruentum

Sodium 

selenate

32 14 days 23-35% reduction

in rate of cell

population growth

10-1,000 Wheeler et

al. 1982

Red alga,

Porphyridium

cruentum

Sodium 

selenate

32 4-5 days 100% mortality 10,000 Wheeler et

al. 1982

Eastern oyster

(adult),

Crassostrea

virginica

Sodium 

selenate

34 14 days No significant

effect on respiration

rate of gill tissue

400 Fowler et al.

1981

Striped bass

(embryo),

Morone saxatilis

Sodium 

selenate

7.2-7.5 4 days 93% successful

hatch and survive

200,000 Klauda

1985a,b

Striped bass

(larva),

Morone saxatilis

Sodium 

selenate

4.0-5.0 4 days LC50 (control

survival= 77%)

13,020 Klauda

1985a,b

Striped bass

(juvenile),

Morone saxatilis

Sodium 

selenate

3.5-5.5 9-65 days Significant

incidence of

development

anomalies of lower

jaw

39-1,360 Klauda

1985a,b

Striped bass

(juvenile),

Morone saxatilis

Sodium 

selenate

3.5-5.5 45 days Significant

incidence of severe

blood cytopathology

1,290 Klauda

1985a,b

a

 Concentration of selenium, not the chemical. Units are μg selenium/L of water unless noted otherwise.

b

 Converted from dry weight to wet weight basis (see Guidelines)

c

 Growth of algae was inhibited

d

 From Smith et al. (1976).

e

 Calculated from the published data using probit analysis and allowing for 8.9% spontaneous mortality.
F-14 Draft November 12, 2004

Other Data - Endangered Species

Two similar studies were conducted in subsequent years, 1996 and 1997, to determine the effects of site water

and site food contaminated with selenium on the endangered species, razorback sucker, Xyrauchen texanus

(Hamilton et al. 2001a,b). Both studies show marked effects on the 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, it was not considered acceptable

data for use in the derivation of the chronic criterion because of inconsistencies between levels of selenium in

the food and larvae and degree and time to response. 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)

This study was initiated with 5-day old razorback sucker larvae spawned from adults which were previously

held (9 months) in three different location 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), Adobe Creek (low level selenium contamination), and North Pond (high level selenium

contamination). The selenium content in the eggs from three Horsethief females ranged from 5.8 to 6.6 μg

Se/g dw, and the selenium content in adult muscle plugs at spawning was from 3.4 to 5.0 μg Se/g dw. The

selenium content in the eggs from three Adobe Creek females ranged from 38.0 to 54.5 μg Se/g dw, and the

selenium content in adult muscle plugs at spawning was from 11.5 to 12.9 μg Se/g dw. The selenium content

in the eggs from three North Pond females ranged from 34.3 to 37.2 μg Se/g dw, and the selenium content in

adult muscle plugs at spawning was from 14.1 to 17.3 μg Se/g dw. The selenium content in the eggs from a

hatchery brood stock female was 7.1 μg Se/g dw, and the selenium content in adult muscle plugs at spawning

ranged from 2.6 to 13.8 μg Se/g dw. The razorback sucker larvae spawned from fish hatchery brood stock

and held in Colorado River (Horsethief) water were used as an additional reference group of test fish. 

The experimental groups were subdivided into those receiving reference water (hatchery water; 24-Road Fish

Hatchery) or site water. They were further subdivided into those receiving a daily ration of reference food

(brine shrimp) or zooplankton 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,
F-15 Draft November 12, 2004

Brood Stock) were exposed to each treatment (2 replicates x 3 spawns x 10 fish/beaker). The larvae were held

in beakers containing 800 mls of test water. Fifty percent of the test water was renewed daily.

Treatment conditions during the 30-day larval study:

Source of Larvae Treatments

Se in 

food

(μg/g dw)

Se in 

water

(μg/L)

Horsethief Adults Reference food: Reference water 2.7 < 1

Reference food: Site water 2.7 0.9

Site food: Reference water 5.6 < 1

Site food: Site water 5.6 0.9

Adobe Creek Adults Reference food: Reference water 2.7 < 1

Reference food: Site water 2.7 5.5

Site food: Reference water 20 < 1

Site food: Site water 20 5.5

North Pond Adults Reference food: Reference water 2.7 < 1

Reference food: Site water 2.7 10.7

Site food: Reference water 39 <1

Site food: Site water 39 10.7

Hatchery raised Adults Reference food: Reference water 2.7 < 1

Reference food: Site water 2.7 0.9

Site food: Reference water 5.6 < 1

Site food: Site water 5.6 0.9

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 were recorded daily. After the 30-day exposure period, the surviving fish were
F-16 Draft November 12, 2004

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 selenium. Specific treatment

conditions were as those described above.

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 μg Se/g 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 μg

Se/g 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 μg Se/g dw (site food-reference water treatment)

and 6.9 μg Se/g dw (site food-site water treatment). 

Several inconsistencies were observed that suggest selenium may not be solely responsible for the effect on

larval survival. Larval survival in the Adobe Creek treatment group exposed to reference water and reference

food was 84 percent, similar to control survival (86-89 percent). The selenium concentration in the larvae

from this Adobe Creek treatment group after 30 days was higher (7.7 μg/g dw) than brood stock fish (5.4 μg

Se/g dw) which had a lower 30-day survival (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.
F-17 Draft November 12, 2004

Evaluation of Contaminant Impacts on Razorback Sucker held in Flooded Bottomland Sites Near Grand

Junction , Colorado - 1997 (Hamilton et al. 2001b)

In a similar 30-day larval study conducted by the authors the in following year (1997), razorback sucker

larvae from a single hatchery brood stock female (11 μg Se/g dw muscle) were subjected to one of 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 mls of test water as before, fifty percent of the test water was renewed daily. Specific

treatment conditions for the 1997 30-day larval study were as follows:

Treatment conditions during the 30-day larval study:

 Water Treatments

Se in 

food

(μg/g dw)

Se in 

water

(μg/L)

Reference food (brine shrimp):

Reference water (24-Road Hatchery)

3.2 < 1

Reference food: Site water (Horsethief) 6.0 1.6

Reference food: Site water (Adobe Creek) 32.4 3.4

Reference food: Site water (North Pond) 52.5 13.3

Horsethief food: Reference water 3.2 < 1

Horsethief food: Site water (Horsethief) 6.0 1.6

Horsethief food: Site water (Adobe Creek) 32.4 3.4

Horsethief food: Site water (North Pond) 52.5 13.3

Adobe Creek food: Reference water 3.2 < 1

Adobe Creek food: Site water (Horsethief) 6.0 1.6

Adobe Creek food: Site water (Adobe Creek) 32.4 3.4

Adobe Creek food: Site water (North Pond) 52.5 13.3

North Pond food: Reference water 3.2 < 1

North Pond food: Site water (Horsethief) 6.0 1.6

North Pond food: Site water (Adobe Creek) 32.4 3.4

North Pond food: Site water (North Pond) 52.5 13.3
F-18 Draft November 12, 2004

In this year’s study, after 30 days of exposure, 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 (only 52 percent). Corresponding selenium concentrations in larval wholebody tissue after 10 days were 6.3, 6.7, and 11 μg Se/g dw, respectively. The average concentrations of

selenium in the water for the three treatments were <1, 1.6, and 13.3 μg Se/L. After 30 days the mean

selenium concentrations in these larvae were 5.2, 5.2, and 16 μg Se/g dw, respectively. Survival was

markedly reduced (0 to 30 percent survival) in the remainder 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, there are several inconsistencies in the results that suggested selenium may not

be 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. 
G-1 Draft November 12, 2004

APPENDIX G

UNUSED DATA
G-2 Draft November 12, 2004

Unused Data

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.

Studies Were Conducted with Species That Are Not Resident in North America

 Ahsanullah and Brand (1985) 

Ahsanullah and Palmer (1980) 

Baker and Davies (1997)

Barghigiani et al. (1993)

Chidambaram and Sastry (1991a,b) 

Congiu et al. (1989) 

Cuvin and Furness (1988) 

Fowler and Benayoun (1976a,b) 

Gaikwad (1989) 

Gotsis (1982) 

Hiraika et al. (1985) 

Juhnke and Ludemann (1978) 

Kitamura (1990) 

Manoharan and Prabakaran (1994) 

Minganti et al. (1994, 1995) 

Niimi and LaHam (1975, 1976) 

Regoli (1998) 

Regoli and Principato (1995)

Rhodes et al. (1994) 

Ringdal and Julshamn (1985)

Rouleau et al. (1992) 

Sastry and Shukla (1994) 

Savant and Nilkanth (1991) 

Shultz and Ito (1979) 

Srivastava and Tyagi (1985)

Takayanagi (2001) 

Tomasik et al. (1995b) 

Tian and Liu (1993) 

Wrench (1978)

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 Reviews Only Contain Data That Have Been Published Elsewhere

Adams and Johnson (1981)

Biddinger and Gloss (1984) 

Bowie et al. (1996) 

Brandao et al. (1992) 

Brooks (1984) 

Burton and Stemmer (1988)

Chapman et al. (1986)

Davies (1978) 

Devillers et al. (1988) 

Eisler (1985) 

Hall and Burton (1982)

Hodson and Hilton (1983)

Hodson et al. (1984) 

Jenkins (1980) 

Kaiser et al. (1997) 

Kay (1984) 

LeBlanc (1984) 

Lemly (1993c, 1996ab, 1997d) 

Lemly and Smith (1987) 

McKee and Wolf (1963) 

National Research Council (1976)

Neuhold (1987)

NCDNR&CD (1986) 

Peterson and Nebeker (1992)

Phillips and Russo (1978) 

Presser (1994) 

Roux et al. (1996) 

Thompson et al. (1972) 

Versar (1975)

. 
G-3 Draft November 12, 2004

Authors Did Not Specify the Oxidation State of Selenium Used in Study

Greenberg and Kopec (1986)

Hutchinson and Stokes (1975)

Kapu and Schaeffer (1991) 

Kramer et al. (1989) 

Mahan et al. (1989)

Rauscher (1988) 

Snell et al. (1991b) 

Selenium Was a Component of an Effluent, Fly Ash, Formulation, Mixture, Sediment or Sludge

Apte et al. (1987) 

Baer et al. (1995) 

Baker et al. (1991) 

Berg et al. (1995) 

Besser et al. (1989) 

Biedlingmaier and Schmidt (1989) 

Bjoernberg (1989) 

Bjoernberg et al. (1988) 

Blockmann et al. (1995) 

Boisson et al. (1989) 

Bondavalli et al. (1996) 

Bowmer et al. (1994) 

Brieger et al. (1992) 

Burton and Pinkney (1984) 

Burton et al. (1983, 1987) 

Cherry et al. (1987) 

Cieminski and Flake (1995) 

Clark et al. (1989) 

Cooke and Lee (1993) 

Cossu et al. (1997) 

Coyle et al. (1993) 

Crane et al. (1992) 

Crock et al. (1992) 

Cushman et al. (1977) 

Davies and Russell (1988) 

de Peyster et al. (1993) 

Dickman and Rygiel (1996)

Dierenfeld et al. (1993) 

Drndarski et al. (1990) 

Eriksson and Forsberg (1992)

Eriksson and Pedros-Alio (1990) 

Fairbrother et al. (1994) 

Fava et al. (1985a,b) 

Feroci et al. (1997) 

Finger and Bulak (1988)

Finley (1985) 

Fisher and Wente (1993)

Fjeld and Rognerud (1993) 

Fletcher et al. (1994) 

Follett (1991) 

Gerhardt (1990) 

Gerhardt et al. (1991) 

Gibbs and Miskiewicz (1995)

Graham et al. (1992) 

Gunderson et al. (1997) 

Hall (1988) 

Hall et al. (1984, 1987, 1988,1992) 

Hamilton et al. (1986, 2000) 

Harrison et al. (1990) 

Hartwell et al. (1987ab, 1988,

1997) 

Hatcher et al. (1992) 

Haynes et al. (1997) 

Hayward et al. (1996) 

Hellou et al. (1996) 

Henebry and Ross (1989) 

Henry et al. (1989, 1990, 1995) 

Hildebrand et al. (1976) 

Hjeltner and Julshman (1992)

Hockett and Mount (1996) 

Hodson (1990) 

Hoffman et al. (1988, 1991)

Homziak et al. (1993) 

Hopkins et al. (2000)

Hothem and Welsh (1994a) 

Jackson (1988) 

Jackson et al. (1990) 

Jacquez et al. (1987) 

Jay and Muncy (1979) 

Jayasekera (1994) 

Jayasekera and Rossbach (1996) 

Jenner and Bowmer (1990) (1992) 

Jenner and Janssen-Mommen

(1989) 

Jin et al. (1997) 

Jorgensen and Heisinger (1987) 

Karlson and Frankenberger (1990) 

Kemble et al. (1994) 

Kenned (1986) 

Kersten et al. (1991) 

King and Cromartie (1986) 

King et al. (1991, 1994) 

Klusek et al. (1993) 

Koh and Harper (1988) 

Koike et al. (1993) 

Krishnaja et al. (1987) 

Kruuk and Conroy (1991) 

Kuehl and Haebler (1995) 

Kuehl et al. (1994) 

Kuss et al. (1995) 

Landau et al. (1985) 

Livingstone et al. (1991) 

Lobel et al. (1990) 
G-4 Draft November 12, 2004

Luoma and Phillips (1988)

Lundquist et al. (1994) 

Lyle (1986) 

MacFarlane et al. (1986) 

Mann and Fyfe (1988) 

Marcogliese et al. (1987) 

Marvin et al. (1997)

Maurer et al (1999) 

McCloskey and Newman (1995)

McCloskey et al. (1995) 

McCrea and Fischer (1986) 

McLean et al. (1991) 

Mehrle et al. (1987) 

Metcalf-Smith (1994) 

Micallef and Tyler (1989) 

Mikac et al. (1985) 

Miles and Tome (1997) 

Miller et al. (1996) 

Misitano and Schiewe (1990)

Moore (1988) 

Munawar and Legner (1993)

Muskett et al. (1985) 

Naddy et al. (1995) 

Nielsen and Bjerregaard (1991) 

Norman et al. (1992)

Nuutinen & Kukkonen (1998) 

Oberbach and Hartfield (1987,

1988) 

Oberbach et al. (1989) 

Ohlendorf et al. (1989, 1990, 1991) 

Olsen and Welsh (1993)

Peters et al.(1999) 

Phillips and Gregory (1980) 

Pratt and Bowers (1990) 

Presser and Ohlendorf (1987)

Prevot and Sayer-Gobillard (1986) 

Pritchard (1997)

Pyleet al. (2001) 

Reash et al. (1988, in press) 

Rhodes and Burke (1996) 

Ribeyre et al. (1995) 

Rice et al. (1995) 

Riggs and Esch (1987) 

Riggs et al. (1987) 

Robertson et al. (1991) 

Roper et al. (1997) 

Russell et al. (1994) 

Ryther et al. (1979) 

Saiki and Jenings (1992) 

Saiki and Ogle (1995) 

Saleh et al. (1988) 

Seelye et al. (1982) 

Sevareid and Ichikawa (1983) 

Skinner (1985) 

Somerville et al. (1987) 

Sorenson and Bauer (1983) 

Specht et al. (1984) 

Steele et al. (1992) 

Stemmer et al. (1990) 

Summers et al. (1995) 

Thomas et al. (1980b)

Timothy et al. (2001) 

Trieff et al. (1995) 

Turgeon and O’Conner (1991) 

Twerdok et al. (1997) 

Ursal (1987) 

Van Metre and Gray (1992) 

Wahl et al. (1994) 

Wandan and Zabik (1996) 

Wang et al. (1992, 1995) 

Welsh (1992) 

Weres et al. (1990) 

White and Geitner (1996)

Wiemeyer et al. (1986) 

Wildhaber and Schmitt (1996) 

Williams et al. (1989) 

Wolfe et al. (1996) 

Wolfenberger (1987) 

Wong and Chau (1988) 

Wong et al. (1982) 

Wu et al. (1997) 

Yamaoka et al. (1994) 

Zagatto et al. (1987) 

Zaidi et al. (1995) 

Zhang et al. (1996)

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) 

Al-Sabti (1994, 1995) 

Arvy et al. (1995) 

Augier et al. (1993) 

Avery et al. (1996) 

Baatrup (1989) 

Baatrup and Dansher (1987)

Baatrup et al. (1986) 

Babich et al. (1986, 1989)
G-5 Draft November 12, 2004

Barrington et al. (1997) 

Becker et al. (1995a,b) 

Bell et al. (1984, 1985, 1986a,b,

1987ab) 

Berges and Harrison (1995)

Blondin et al. (1988) 

Boisson et al. (1996) 

Bottino et al. (1984) 

Braddon (1982) 

Braddon-Galloway and Balthrop

(1985) 

Bradford et al. (1994a,b) 

Brandt et al. (1990) 

Byl et al. (1994) 

Chandy and Patel (1985) 

Chen et al. (1997) 

Cheng et al. (1993) 

Christensen and Tucker (1976) 

Dabbert and Powell (1993)

DeQuiroga et al. (1989) 

Dierickx (1993) 

Dietrich et al. (1987) 

DiIlio et al. (1986) 

Doyotte et al. (1997) 

Drotar et al. (1987) 

Dubois and Callard (1993) 

Ebringer et al. (1996) 

Engberg and Borsting (1994)

Engberg et al. (1993) 

Eun et al. (1993) 

Foltinova and Gajdosova (1993) 

Foltinova et al. (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 et al. (1987)

 Henny and Bennett (1990) 

Hoffman and Heinz (1988, 1998) 

Hoffman et al. (1989, 1998)

Hontela et al. (1995) 

Hoglund (1991) 

Hsu et al. (1995) 

Hsu and Goetz (1992) 

Ishikawa et al. (1987) 

James et al. (1993) 

Jovanovic et al. (1995, 1997) 

Kai et al. (1995) 

Kedziroski et al. (1996) 

Kelley et al. (1987) 

Kralj and Stunja (1994) 

Lalitha and Rani (1995) 

Lan et al. (1995) 

Lemaire et al. (1993) 

Livingstone et al. (1992) 

Low and Sin (1995, 1996) 

Micallef and Tyler (1990)

Montagnese et al. (1993) 

Murata et al. (1996) 

Nakonieczny (1993) 

Neuhierl and Boeck (1996) 

Nigro et al (1992, 1994) 

Norheim and Borch-Iohnsen (1990) 

Norheim et al. (1991) 

O’Brien et al. (1995) 

Olson and Christensen (1980) 

Overbaugh and Fall (1985)

Palmisano et al. (1995) 

Patel et al. (1990) 

Patel and Chandy (1987) 

Perez et al. (1990) 

Perez-Trigo et al. (1995) 

Phadnis et al. (1988) 

Price and Harrison (1988) 

Rady et al. (1992)

Rani and Lalitha (1996) 

Regoli et al. (1997) 

Schmidt et al. (1985) 

Schmitt et al. (1993) 

Segner et al. (1994) 

Sen et al. (1995) 

Shigeoka et al. (1990, 1991)

Siwicki et al. (1994) 

Srivastava and Srivastava (1995) 

Sun et al. (1995) 

Takeda et al. (1992a,b,(1993,

1997) 

Treuhardt (1992) 

Vazquez et al. (1994) 

Veena et al. (1997) 

Wise et al. (1993a,b) 

Wong and Oliveira (1991) 

Yokota et al. (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 ? g/L) of EDTA (Riedel and Sanders (1996). 
G-6 Draft November 12, 2004

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. 

Data Were Not Used When the Organisms Were Exposed to Selenium by Food or by Gavage or Injection

Frankenberger and Engberg (in

press)

Hamilton (1999)

Hamilton and Lemly (1999)

Heinz and Sanderson (1990) 

Heinz et al. (1990, 1996)

Hilton et al. (1982) 

Hoffman and Heinz (1988) 

Hoffman et al. (1991,

1992a,b,1996)

Huerkamp et al. (1988) 

Julshamn et al. (1990) 

Kleinow (1984) 

Kleinow and Brooks (1986a,b)

Lemly (1996, 1997, 1999) 

Lorentzen et al. (1994) 

Maage and Waagboe (1990) 

Malchow et al. (1995)

Paripatananontand Lovell(1997) 

Sheline and Schmidt-Nielson

(1977) 

Stanley et al. (1994, 1996)

Wiemeyer and Hoffman (1996) 

Wilson et al. (1997)

BCFs and BAFs from laboratory tests were not used when the tests were static or when the concentration

of selenium in the test solution was not adequately measured or varied too much (Nassos et al. 1980; Ornes

et al. 1991; Riedel et al. 1991; Sharma and Davis 1980; Vandermeulen and Foda 1988). 

Selenium Concentrations Reported in Wild Aquatic Organisms Were Insufficient to Calculate BAF

Abdel-Moati and Atta (1991)

Adeloju and Young (1994) 

Aguirre et al. (1994) 

Akesson and Srikumar (1994) 

Aksnes et al. (1983) 

Allen and Wilson (1990) 

Ambulkar et al. (1995) 

Amiard et al. (1991, 1993) 

Andersen and Depledge (1997) 

Andreev and Simeonov (1992) 

Angulo (1996) 

Arrula et al. (1996) 

Arway (1988) 

Ashton (1991)

Augier et al. (1991, 1993, 1995a,b) 

Augspurger et al. (1998) 

Avery et al. (1996) 

Badsha and Goldspink (1988)
G-7 Draft November 12, 2004

Baines and Fisher (2001) 

Baldwin and Maher (1997) 

Baldwin et al. (1996) 

Barghigiani (1993) 

Barghigiani et al. (1991) 

Baron et al. (1997) 

Batley (1987) 

Baumann and Gillespie (1986) 

Baumann and May (1984) 

Beal (1974) 

Beck et al. (1997) 

Beland et al. (1993) 

Beliaeff et al. (1997) 

Bell and Cowey (1989) 

Benemariya et al. (1991) 

Berry et al. (1997)

Bertram et al. (1986) 

Besser et al. (1994, 1993) 

Birkner (1978) 

Boisson and Romeo (1996)

Bowerman et al. (1994) 

Braune et a. (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 et al. (1992a,b,c,1993,

1994a,b) 

Byrne and DeLeon (1986) 

Byrne et al. (1985) 

Cantillo et al. (1997) 

Capar and Yess (1996) 

Capelli et al. (1987, 1991) 

Cappon (1984) 

Cappon and Smith (1981)

(1982a,b)

Cardellicchio (1995) 

Carell et al. (1987) 

Carter and Porter (1997) 

Caurant et al. (1994, 1996) 

Chau and Riley (1965) 

Chiang et al. (1994) 

Chou and Uthe (1991) 

Chvojka (1988) 

Chvojka et al. (1990) 

Clifford and Harrison (1988)

Collins (1992) 

Combs et al. (1996) 

Cosson et al. (1988) 

Courtney et al. (1994) 

Crowys et al. (1994)

Crutchfield (2000) 

Cumbie and Van Horn (1978) 

Currey et al. (1992) 

Custer and Hohman (1994) 

Custer and Mitchell (1991, 1993) 

Custer et al. (1997)

Dabeka and McKenzie (1991) 

Davoren (1986) 

Deaker and Maher (1997) 

Demon et al. (1988) 

Dietz et al. (1995, 1996) 

Doherty et al. (1993) 

Elliott and Scheuhammer (1997) 

Eriksson et al. (1989) 

Evans et al. (1993) 

Felton and Mathews (1990) 

Felton et al. (1994) 

Fitzsimmons et al. (1995) 

Focardi et al. (1985, 1988) 

Fowler (1986) 

Fowler et al. (1975, 1985) 

France (1987) 

Friberg (1988) 

Froslie et al. (1985, 1987)

Gabrashanske and Daskalova

(1985) 

Gabrashanska and Nedeva (1994) 

Galgan and Frank (1995)

Garcia - Hermandez et al. (2000)

Giardina et al. (1997) 

Gillespie and Baumann (1986) 

Gochfeld (1997) 

Goede (1985, 1991, 1993a,b)

Goede et al. (1989, 1993) 

Goede and DeBruin (1984, 1985) 

Goede and Wolterbeek (1993,

1994a,b) 

Gras et al. (1992) 

Greig and Jones (1976) 

Gutenmann et al. (1988) 

Gutierrez-Galindo et al. (1994) 

Guven et al. (1992) 

Halbrook et al. (1996)

Hall and Fisher (1985) 

Hamilton and Waddell (1994) 

Hamilton and Wiedmeyer (1990) 

Hansen et al. (1990) 

Hardiman and Pearson (1995) 

Hargrave et al. (1992) 

Harrison and Klaverkamp (1990) 

Hasunuma et al. (1993) 

Haynes et al. (1995) 

Hein et al. (1994) 

Heiny and Tate (1997) 

Heinz (1993a) 

Heinz and Fitzgerald (1993a,b) 

Heit (1985) 

Heit and Klusek (1985) 

Heit et al. (1980, 1989) 
G-8 Draft November 12, 2004

Hellou et al. (1992a,b) (1996a,b) 

Henny and Herron (1989) 

Hodge et al. (1996)

Hilton et al. (1982) 

Honda et al. (1986) 

Hothem and Ohlendorf (1989) 

Hothem and Welsh (1994b) 

Hothem and Zador (1995) 

Hothem et al. (1995) 

Houpt et al. (1988) 

Hunter et al. (1995, 1997) 

Ibrahim and Farrag (1992) 

Ibrahim and Mat (1995) 

Ishikawa et al. (1993) 

Itano et al. (1984, 1985a,b) 

Jarman et al. (1996) 

Johns et al. (1988) 

Johnson (1987) 

Jop et al. (1997) 

Jorhem et al. (1994)

Julshamn et al. (1987) 

Kai et al. (1986a,b, 1988, 1992a,b,

1996) 

Kaiser et al. (1979) 

Kalas et al. (1995) 

Kidwell et al. (1995) 

Koeman et al. (1973) 

Kovacs et al. (1984) 

Krogh and Scanes (1997)

Krushevska et al. (1996)

Lakshmanan and Stephen (1994) 

Lalitha et al. (1994) 

LamLeung et al. (1991) 

Lan et al. (1994a,b) 

Langlois and Langis (1995) 

Larsen and Stuerup (1994) 

Larsen et al. (1997) 

Lauchli (1993) 

Law et al. (1996) 

Lee and Fisher (1992a,b, 1993) 

Leighton and Wobeser (1994) 

Leland and Scudder (1990) 

Lemly (1985a, 1994) 

Leonzio et al. (1986, 1989, 1992) 

Leskinen et al. (1986) 

Li et al. (1996) 

Lie et al. (1994) 

Liu et al. (1987) 

Lizama et al. (1989) 

Lobel et al. (1989, 1991, 1992a,b) 

Lonzarich et al. (1992) 

Lourdes et al. (1990) 

Lowe et al. (1985) 

Lucas et al. (1970) 

Lytle and Lytle (1982) 

Mackey et al. (1996) 

Maher (1987) 

Maher et al. (1992, 1997) 

Mann et al. (1988)

Mason et al. (2000) 

Masuzawa et al. (1988) 

Matsumoto (1991)

Maven et al. (1995) 

May and McKinney (1981)

Mcdowell et al. (1995) 

McKenzie-Parnell et al. (1988) 

Meador et al. (1993) 

Mehrle et al. (1982) 

Meltzer et al. (1993) 

Metcalfe-Smith et al. (1992, 1996) 

Michot et al. (1994) 

Mills et al. (1993) 

Moharram et al. (1987) 

Moller (1996) 

Mora and Anderson (1995) 

Morera et al. (1997) 

Muir et al. (1988) 

Mutanen et al. (1986) 

Nadkarni and Primack (1993)

Nakamoto and Hassler (1992) 

Narasaki and Cao (1996) 

Navarrete et al. (1990) 

Nettleton et al. (1990) 

Nicola et al. (1987) 

Nielsen and Dietz (1990) 

Norheim (1987) 

Norheim et al. (1992)

Norrgren et al. (1993) 

Norstrom et al. (1986) 

O’Conner (1996) 

O’Shea et al. (1984) 

Ober et al. (1987) 

Oehlenschlager (1997) 

Ohlendorf (1986) 

Ohlendorf and Harrison (1986) 

Ohlendorf and Maron (1990) 

Ohlendorf et al. (1986a,b, 1987,

1988a,b) 

Okazaki and Panietz (1981)

Ostapczuk et al. (1997) 

Pakkala et al. (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) 

Park et al. (1994) 

Paveglio et al. (1994) 

Payer and Runkel (1978) 

Payer et al. (1976) 

Pennington et al. (1982) 
G-9 Draft November 12, 2004

Presley et al. (1990) 

Quevauviller et al. (1993a,b)

Ramos et al. (1992) 

Rao et al. (1996) 

Reinfelder and Fisher (1991)

Reinfelder et al. (1993, 1998) 

Renzoni et al. (1986) 

Riget et al. (1996) 

Risenhoover (1989)

Roditi (2000) 

Roux et al. (1994) 

Ruelle and Keenlyne (1993) 

Sager and Cofield (1984) 

Saiki (1986 ab, 1987, 1990) 

Saiki and Lowe (1987) 

Saiki and May (1988) 

Saiki and Palawski (1990) 

Saiki et al. (1992, 1993) 

Sanders and Gilmour (1994) 

Scanes (1997) 

Scheuhammer et al. (1988) 

Schantz et al. (1997) 

Schmitt and Brumbaugh (1990) 

Schramel and Xu (1991) 

Schuler et al. (1990) 

Scott and Latshaw (1993) 

Secor et al. (1993) 

Seelye et al. (1982) 

Sharif et al. (1993) 

Shen et al. (1997) 

Shirasaki et al. (1996) 

Shultz and Ito (1979) 

Simopoulos (1997) 

Skaare et al. (1990, 1994)

Smith and Flegal (1989) 

Smith et al. (1992) 

Sorensen (1988) 

Sorensen and Bauer (1984a,b)

Sorensen and Bjerregaard (1991) 

Sorensen et al. (1982, 1983, 1984)

Southworth et al. (2000)

Sparling and Lowe (1996) 

Speyer (1980) 

Steimle et al. (1994) 

Stoeppler et al. (1988) 

Stone et al. (1988) 

Stripp et al. (1990) 

Sundarrao et al. (1991) (1992) 

Svensson et al. (1992) 

Tabaka et al. (1996) 

Talbot and Chang (1987) 

Tallandini et al. (1996) 

Tan and Marshall (1997)

Tang et al. (1997) 

Tao et al. (1993) 

Teherani (1987) 

Teigen et al. (1993)

Thomas et al. (1999) 

Tilbury et al. (1997) 

Topcuoglu et al. (1990) 

TranVan and Teherani (1988) 

Trocine and Trefry (1996) 

Uthe and Bigh (1971) 

Vanderstoep et al. (1990) 

Varanasi et al. (1993, 1994)

Vitaliano and Zdanowicz (1992) 

Vlieg (1990)

Vlieg et al. (1993) 

Vos et al. (1986) 

Waddell and May (1995)

Wagemann (1988) 

Wagemann and Stewart (1994) 

Wagemann et al. (1988) (1996)

Walsh et al. (1977) 

Wang (1996) 

Ward and Flick (1990) 

Warren et al. (1990) 

Weber (1985) 

Welsh and Maughan (1994) 

Wen et al. (1997) 

Wenzel and Gabrielsen (1995) 

Whyte and Boutillier (1991)

Williams et al. (1994) 

Wilson et al. (1992, 1997) 

Winger and Andreasen (1985) 

Winger et al. (1984, 1990) 

Woock and Summers (1984) 

Wren et al. (1987) 

Wu and Huang (1991) 

Yamaoka et al. (1996) 

Yamazaki et al. (1996)

Yoshida and Yasumoto (1987) 

Zatta et al. (1985) 

Zeisler et al. (1988, 1993) 

Zhou and Liu (1997)
H-1 Draft November 12, 2004

APPENDIX H

DATA USED IN REGRESSION ANALYSIS OF SELENIUM IN WHOLE-BODY 

FISH TISSUE WITH SELENIUM IN MUSCLE, OVARY AND LIVER TISSUES
H-2 Draft November 12, 2004

DATA USED IN REGRESSION ANALYSIS OF SELENIUM IN WHOLE-BODY 

FISH TISSUE WITH SELENIUM IN MUSCLE, OVARY AND LIVER TISSUES

Quantile regression was used to estimate median concentrations of selenium in the whole body as a

function of selenium concentration in selected tissues (Tables H-2, H-3, H-4). Only data where

organisms were exposed to selenium in water and in diet or in only diet were considered for analysis.

Quantile regression fits a curve to the data such that a selected proportion t (the quantile) of observations

are below and the complementary fraction 1- t is above it (Koenker and Basset 1978). Estimates of

model parameters minimize the sum of weighted absolute deviations. In contrast, ordinary least squares

minimize the sum of squared deviations. Least absolute deviation is less sensitive to outliers than least

squares (Birkes and Dodge 1993). Other desired properties of quantile regression include: it is

equivariant to scale changes, location shift, and monotonic transformations (Koenker and Basset 1978,

Cade et al. 1999). Furthermore, with rank-score statistics it is possible to test hypotheses and build

confidence intervals for parameters of linear models fit to data with heteroscedastic errors (Koenker

1994, Koenker and Machado 1999). The rank-score test does not have to assume homogeneous error

distributions because the statistic is based on signs of residuals and not their size (Koenker and Machado

1999). For introductory presentations of quantile regression see Cade et al. (1999), Koenker and Hallock

(2001), and Cade & Noon (2003). All quantile regressions reported here were performed using the R

software (Ihaka & Gentleman 1996) version 1.8.0.

As the exact form of the relationship between selenium concentrations in the whole body ([Se]WB) and in

tissues ([Se]Tissue) is not known, we considered three candidate models : 

I) [Se]WB = a. 

II) [Se]WB = a + b [Se]Tissue and 

III) [Se]WB = exp(a + b ln([Se]Tissue))

where a and b are the model parameters we wish to estimate. Model (I) implicitly assumes that selenium

concentrations in the whole body are independent of selenium concentrations in liver, muscle, or ovary

tissues. Model (II) projects selenium concentrations in the whole body as a linear function of selenium

concentrations in a tissue. Model (III) estimates selenium concentrations in the whole body as an

exponential function of the logarithm of selenium concentrations in a tissue. This model is derived from

the assumption of a linear relationship between the natural logarithms of [Se]WB and [Se]Tissue.

Selection of the best model(s) considers both the fit and number of parameters. Models with greater

number of parameters generally fit the data better, but such reduction in bias is invariably associated with

an increase in variance of parameter estimates (Burnham and Anderson 2002). Model selection methods

attempt to find a parsimonious model with the proper tradeoff between bias and variance. We apply the

information theoretic approach for model selection (Burnham and Anderson 2002). It is based on the

Kullback-Leibler information, I(f,g), which expresses the information lost when model g is used to

estimate the full reality f. Obviously, the full reality is never known, but an estimate of the relative

distance from reality can be estimated by the Akaike Information Criterion (AIC, Akaike 1973)

AIC = -2 ln(‹(parameters|data)) + 2k
H-3 Draft November 12, 2004

where k is the number of parameters in the model and ‹(parameters|data) is the maximized likelihood of

parameter estimates for the available data.. The AIC is a poor estimator of I(f,g) when n/k < 40 (n is the

sample size). In such instances, a second-order version of AIC, AICc, is recommended (Hurvich and Tsai

1989):

AICc

 = -2 ln(‹(parameters|data)) + 2k

Hurvich and Tsai (1990) demonstrated that the modified version of AICc for least absolute

deviation(L1AICc) provides an unbiased estimator for the Kullback-Leibler information, but the small

sample criterion for normal least squares regression, which is less computationally demanding, performs

equally well 

 

where F

 2 is estimated as the sum of squared residuals divided by n. For the least absolute deviation

regression, F

 2 is estimated as (SWAD/n)

2

, thus AICc is computed by the expression

The AIC and AICc

 are used to rank candidate models. Comparisons among the M ranked

candidates are based on the Akaike weight (w), which represents the likelihood of a model given the data 

where )iis the difference in AIC (AICc) between model i and the model with the lowest AIC (AICc)

value. Weights for all candidate models sum to 1. For each model, we computed the sum of weighted

absolute deviations (SWAD), AICc and the Akaike weight (Table H-1). 

The linear model (II) was selected the best among the three candidate functions for projecting

concentrations of selenium in the whole body as a function of selenium concentrations in the liver (Table

H-1). The exponential model (III) was selected the best for projections based on concentrations of

selenium in muscles and ovaries. However, fits of models II and III to ovary data had similar weights. As

the best model may not explain much of the observed variation in the data, we calculated coefficients of

determination (R1), defined as

R

1

 = 1 - (SAF/SAR)
H-4 Draft November 12, 2004

where SAF and SAR are the sum of weighted absolute deviations for the full and reduced models,

respectively (Cade and Richards 1996). Coefficients of determination for models II and III were also

very similar, suggesting that both models are equally effective in predicting concentrations of selenium in

the whole body as a function of selenium concentrations in ovaries. With such knowledge, we opted to

use the linear model (II) because it is easier to compute. The exponential model for muscle presented the

highest coefficient of determination (0.77), indicating that samples of selenium concentrations from this

tissue are more effective predictors than samples from liver and ovaries. The fitted quantile regression

curves are shown in figure 5 of the selenium document.

TableH- 1. Number of parameters (k), sum of weighted absolute deviations (SWAD), second-order

Akaike Information Criterion (AICc), differences between the model AICc and the lowest AICc of all

candidate models (Delta), weight (w), rank (by weight), and coefficient of determination (R1) for three

candidate models to project selenium concentrations in the whole body as a function of selenium

concentrations in a selected tissue.

Tissue: Muscle (n = 21)

Model k SWAD AICc Delta Weight Rank R1

[Se]WB = a 2 66.00 52.76 59.20 1.27e-13 3

[Se]WB = a + b [Se]Tissue 3 16.84 -1.85 4.59 9.17e-02 2 0.74

[Se]WB = exp(a + b*ln([Se]Tissue)) 3 15.10 -6.43 0.00 9.08e-01 1 0.77

Tissue: Ovary (n = 23)

Model k SWAD AICc Delta Weight Rank R1

[Se]WB = a 2 73.95 58.32 46.89 3.31e-11 3

[Se]WB = a + b [Se]Tissue 3 25.20 11.46 0.03 4.97e-01 2 0.66

[Se]WB = exp(a + b*ln([Se]Tissue)) 3 25.18 11.43 0.00 5.03e-01 1 0.66

Tissue: Liver (n = 26)

Model k SWAD AICc Delta Weight Rank R1

[Se]WB = a 2 41.05 28.27 22.81 1.11e-05 3

[Se]WB = a + b [Se]Tissue 3 25.20 5.46 0.00 9.99e-01 1 0.39

[Se]WB = exp(a + bln([Se]Tissue)) 3 40.83 30.56 25.10 3.54e-06 2 0.01
H-5 Draft November 12, 2004

Table H-1. Whole body vs muscle

Se in tissue, μg/g dw

reference species site/treatment muscle whole body

Hermanutz et al. 1996 bluegill I control-down 2.05 1.95

bluegill I 10 μg/L-down 20.55 22.85

bluegill II control-up 1.9 2.45

bluegill II control-down 2.25 1.95

bluegill II 2.5 μg/L-up 3.5 3.5

bluegill II 2.5 μg/L-down 6.9 6.15

bluegill II 10 μg/L-up 17.55 15.45

bluegill II 10 μg/L-down 44.7 26.45

bluegill II rec 30-up 12.45 11.85

bluegill II rec 30-down 39.6 30.6

bluegill III control-up 3.35 3.35

bluegill III control-down 3.2 2.3

bluegill III rec 2.5-up 5.25 6.3

bluegill III rec 2.5-down 6.1 5.3

bluegill III rec 10-up 12.45 12

bluegill III rec 10-down 18.6 13

bluegill III rec 30-up 7.75 8.35

bluegill III rec 30-down 15.05 17.35

Garcia-Hernandez 2000 tilapia Cienega de Santa Cl 3.5 3

carp Cienega de Santa Cl 4.6 3.3

LM bass Cienega de Santa Cl 5.4 5.1
H-6 Draft November 12, 2004

Table H-2. Whole Body vs Ovary

Se in tissue, μg/g dw

reference species site/treatment ovary whole body

Coyle 1993 bluegill control 2.1 0.9

bluegill control + water Se 2.1 0.9

bluegill 4.6 μg/g diet 8.3 2.9

bluegill 8.4 μg/g diet 12.5 4.9

bluegill 16.8 μg/g diet 25 7.2

bluegill 33.3 μg/g diet 41 16

Hermanutz et al. 1996 bluegill I control-down 0.35 1.95

bluegill I 10 μg/L-down 20.05 22.85

bluegill II control-up 5.25 2.45

bluegill II control-down 3.85 1.95

bluegill II 2.5 μg/L-up 10.1 3.5

bluegill II 2.5 μg/L-down 12.35 6.15

bluegill II 10 μg/L-up 34.8 15.45

bluegill II 10 μg/L-down 50.5 26.45

bluegill II rec 30-up 29.35 11.85

bluegill II rec 30-down 66 30.6

bluegill III control-down 5.3 2.3

bluegill III rec 2.5-up 8.4 6.3

bluegill III rec 2.5-down 9.5 5.3

bluegill III rec 10-up 31.15 12

bluegill III rec 10-down 19.55 13

bluegill III rec 30-up 17.85 8.35

bluegill III rec 30-down 19.1 17.35
H-7 Draft November 12, 2004

Table H-3. Whole body vs liver

Se in tissue, μg/g dw

reference species site/treatment liver whole body

Bryson 1985-84 bluegill control 3.9 0.45

bluegill Se-plankton diet 9.1 2.35

bluegill Selenite diet 11 1.21

bluegill Se-cysteine diet 9.23 2.16

bluegill Se-cysteine 2X diet 16.33 3.74

bluegill Se-methionine diet 10.85 2.46

Hermanutz et al. 1996 bluegill I control-down 5.4 1.95

bluegill I 10 μg/L-down 36.05 22.85

bluegill II control-up 13.2 2.45

bluegill II control-down 7.2 1.95

bluegill II 2.5 μg/L-up 29.2 3.5

bluegill II 2.5 μg/L-down 26.45 6.15

bluegill II 10 μg/L-up 119 15.45

bluegill II 10 μg/L-down 68.5 26.45

bluegill II rec 30-up 64 11.85

bluegill II rec 30-down 100.5 30.6

bluegill III control-up 9.95 3.35

bluegill III control-down 9.4 2.3

bluegill III rec 2.5-up 13.85 6.3

bluegill III rec 2.5-down 16.3 5.3

bluegill III rec 10-up 33.25 12

bluegill III rec 10-down 37.15 13

bluegill III rec 30-up 21 8.35

bluegill III rec 30-down 31.9 17.35

Garcia-Hernandez 2000 carp Cienega de Santa Cl 8.2 3.3

LM bass Cienega de Santa Cl 4.7 5.1
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APPENDIX I

SUMMARIES OF CHRONIC STUDIES CONSIDERED FOR FCV DERIVATION
I-2 Draft November 12, 2004

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: Rotifer (Brachionus calyciflorus), and fathead minnow (Pimephales promelas)

12 to 24 hr-old at start.

Exposure Route: 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 μg 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 μg total Se/L); rotifers were

exposed from the water (1, 108.1, 204.9, 393.0 μg total Se/L) and the green alga

as food (2.5, 33, 40, 50 μg Se/g dry wt.); and the fathead minnow were exposed

from water (1, 108.1, 204.9, 393.0 μg total Se/L) and the rotifer as food (2.5, 47,

53, 60 μg Se/g dry wt.).

Test Duration: 25 days

Study Design: 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 × 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 have 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
I-3 Draft November 12, 2004

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. 

Additional measurements were made in the 200 μg 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.

Effects Data: 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 μg/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, μg/L Se in diet, μg/g dw Se in rotifer tissue,

μg/g dw

rotifer biomass, mg/ml

dw

1 2.5 2.5 0.028

108.1 33 40 0.025

202.4 40 54 0.011

393 50 75 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.

Effect of Dietary and Waterborne Selenium on Larval Fathead Minnows after 8 Days Exposure

Se in water, μg/L Se in diet, μg/g dw Se in fathead minnow

tissue, μg/g dw

average fish weight,

mg dw

1 2.5 2.5 0.8

108.1 47 45 0.7

202.4 53 75 0.4

393 60 73 0.2
I-4 Draft November 12, 2004

Chronic Value:

Rotifers 42.36 μg Se/g dw (EC20)

Fish < 73 μg Se/g 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.
I-5 Draft November 12, 2004

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: Chinook salmon (Oncorhynchus tshawytscha Walbaum; swim-up larvae)

Exposure Route: 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 μg Se/g 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 μg Se/g 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 μg Se/g dw), termed SeMet diet.

Dietary Treatments: Each selenium diet was formulated to contain about 36 μg Se/g 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 μg Se/g dw).

Test Duration: 90 days

Study Design: 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 (dry wt. 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 μg/L) in all dietary selenium exposure

concentrations.
I-6 Draft November 12, 2004

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, μg/g dw Se in chinook salmon,

μg/g dw

mean larval weight, g survival, %

1 0.9 3.35 99

3.2 3.3 2.68 97.3

5.3 4.5 2.76 93

9.6 8.4 2.8 95

18.2 13.3 2.62 92.4

35.4 29.4 1.4 89

Effect of Seleno-methionine Diet on Growth and Survival of Chinook Salmon Larvae after 60 Days

Se in diet, μg/g dw Se in chinook salmon,

μg/g dw

mean larval weight, g survival, %

1 0.9 3.35 99

3.2 2 3.08 100

5.3 3.1 3.22 95

9.6 5.3 3.07 94.1

18.2 10.4 2.61 92.4

35.4 23.4 1.25 62.5
I-7 Draft November 12, 2004

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

EC20 values

Survival

(after 60 d of exposure)

Growth

(after 60 d of exposure)

Tissue Se

(μg/g dw)

Tissue Se 

(μg/g dw)

SLD NAa

15.74

SeMet NAa10.47

a The EC20 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. 
I-8 Draft November 12, 2004
I-9 Draft November 12, 2004
I-10 Draft November 12, 2004

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: Rainbow trout (Oncorhynchus mykiss; juvenile; approx. 0.6 g each)

Exposure Route: 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 cerelose 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.

Test Treatments: The two diets were supplemented with selenium (as sodium selenite) at the rate

of 0, 5, or 10 μg/g 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 μg/g dw, and the measured concentrations of

selenium in the high carbohydrate diet were: 0.7 (control), 6.6, and 11.8 μg/g

dw. The tanks received a continuous flow of water with a flow rate of 3-4 Liters

per minute.

Test Duration: 16 weeks

Study Design: 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. 

Effects Data: 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
I-11 Draft November 12, 2004

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.

Effect of Selenium in Low carbohydrate Diet to Rainbow Trout

Se in diet, μg/g dw Se in trout liver, μg/g dw trout weight, kg/100 fish

0.6 0.8 3.3

6.6 38.3 3.3

11.4 49.3 1.8

Effect of Selenium in High carbohydrate Diet to Rainbow Trout

Se in diet, μg/g dw Se in trout liver, μg/g dw trout weight, kg/100 fish

0.7 0.6 2.7

6.6 21.0 2.3

11.8 71.7 1.4

Chronic Value: 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) μg/g dw, or 43.45 μg/g dw. The MATC

estimated for growth of rainbow trout relative to final concentration of selenium

in liver tissue of trout reared on the high carbohydrate diet is the GM of 21.0

(NOAEC) and 71.7 (LOAEC) μg/g dw, or 38.80 μg/g dw. Using equation III in

the text to convert this selenium concentration in liver tissue to a concentration

of selenium in whole-body, the MATC for rainbow trout exposed to selenium in

food with low carbohydrate content becomes 13.08 μg Se/g dw., whereas the

MATC for rainbow trout exposed to selenium in food with high carbohydrate

content becomes 11.65 μg Se/g dw. The latter value is selected as the chronic

value for the study.

EC20 values could not be determined for this study. Data did not meet minimum

requirements for analysis.
I-12 Draft November 12, 2004

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: Rainbow trout (Oncorhynchus mykiss; juvenile; approx. 0.6 g each)

Exposure Route: 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.

Test Treatments: The test diet was supplemented with selenium (as sodium selenite) at the rate of

0, 5, or 10 μg/g 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 μg/g dw. The tanks received

a continuous flow of water with a flow rate of 3-4 Liters per minute.

Test Duration: 16 to 20 weeks

Study Design: 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. 

Effects Data: 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. 

Chronic Value: 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) μg/g dw, or 43.45 μg/g dw. Using equation

III to convert the selenium concentration in liver tissue to a concentration of

selenium in whole-body, the MATC becomes 13.08 μg/g dw.

EC20 values could not be determined for this study. Data did not meet minimum

requirements for analysis.
I-13 Draft November 12, 2004

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: Rainbow trout (Oncorhynchus mykiss; juvenile; approx. 1.28 g each)

Exposure Route: Dietary only

A casien-torula yeast diet was formulated to contain geometrically increasing

levels of selenium from 0 to 15 μg/g dw. The selenium was supplemented as

sodium selenite which was mixed with cellulose and then added to the diet as a

selenium premix.

Test Duration: 20 weeks

Study Design: 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 μg/g dw. The tanks received a continuous flow of

dechlorinated tap water from the City of Burlinton, Ontario municipal water

supply. The waterborne selenium content of this water was 0.4 μg/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.

Effects Data: 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 μg/g 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. 
I-14 Draft November 12, 2004

Effects on Juvenile Rainbow Trout 

Se in diet, μg/g dw Se in Liver, μg/g dw weight, g/fish mortality*

0.07 0.6 3.2 0

0.15 0.95 3.5 0

0.38 2.4 3.7 0.6

1.25 11 4.1 0.6

3.67 40 4.1 0

13.06 100 1.4 10.7

*expressed as number per 10,000 fish-days

Chronic Value: An MATC was preferred over regression analysis because of the large standard

error associated with the EC20 value. The MATC for the growth and survival of

juvenile trout based on selenium in liver tissue is the GM of the NOAEC (40

μg/g dw) and the LOAEC (100 μg/g dw), or 63.25 μg Se/g dw. Using the

equation III in the text to convert the selenium concentration in liver tissue to a

concentration of selenium in whole-body tissue, the MATC becomes 19.16 μg/g

dw. 
I-15 Draft November 12, 2004

Holm, J. 2002. Sublethal effects of selenium on rainbow trout (Oncorhynchus mykiss) and brook trout

(Salvelinus fontinalis). 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 (Salvelinus fontinalis) 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.

Test Organism: Rainbow trout (Oncorhynchus mykiss; spawning adults) and brook trout

(Salvelinus fontinalis; spawning adults) 

Exposure Route: dietary and waterborne - field exposure

Total selenium concentrations measured at the high selenium site ranged from 6

to 32 μg/L. Selenium was not measured at the reference streams; selenium

concentrations at reference locations in the area ranged from <0.5 to 2.2 μg/L.

Study Design: Spawning fish were collected at low selenium or reference streams (Deerlick

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 both 2000

and 2001. 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..

Effects Data : 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

set with each year (within streams approach). This approach, which results in

an EC20 value if the data meet the model assumptions, is explained in the

Calculations of Chronic Values section of the text. 
I-16 Draft November 12, 2004

Between streams approach: For both rainbow and brook trout embryos, there

were no significant differences in fertilization, time to hatch and mortality

between the streams with elevated selenium and the reference streams in both

2000 and 2001. The frequency of embryonic effects were significantly greater in

the high selenium stream (Luscar Creek) in 2000. Rainbow trout embryos from

Luscar Creek had a greater frequency of craniofacial, skeletal and finfold

deformities and edema; whereas brook trout from Luscar Creek had a greater

frequency of only craniofacial deformities (see Holm Tables 1 and 2 below). In

2001, however, there were no significant differences in embryonic deformities

between Luscar Creek and reference streams for both species of trout. The only

difference observed in 2001 was a greater frequency of finfold deformities in

brook trout collected from Gregg Creek (intermediate selenium levels) relative to

the reference stream (see Holm table 2 below). All other embryonic

measurements in 2001 were not significantly different between streams with

elevated selenium and reference streams. When the data for both years were

pooled, no significant effects were observed in embryos obtained from rainbow

and brook trout collected in Luscar Creek relative to reference streams (see

Holm Table 3). 

Within streams approach: EC20 values could not be calculated for total

deformities or edema for the 2000 rainbow trout data because a logistic curve

could not be fitted to the data (see Holm Figures 1 and 2). For the 2001 data,

EC20 values could not be computed for edema and skeletal and finfold

deformities for rainbow trout data because a logistic curve could not be fitted to

the data (see Holm Figures 3 and 4). Craniofacial deformities in the rainbow

embryo as a function of selenium in egg ww (2001 data) was fitted to a logistic

curve from which an EC20 value was calculated (see Holm Figure 5). The brook

trout data for 2000 and 2001 were not suitable for fitting logistic curves (see

Holm Figure 6).
I-17 Draft November 12, 2004

Holm Table 1 

Mean embryo-larval parameters for rainbow trout 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 two consecutive years (mean ± SE). Values that are

significantly different at " = 0.05 are marked with different letters. (Table modified from Holm 2002)

Measurement

2000 2001

Luscar Deerlick Luscar Gregg Deerlick Wampus

Se, egg, μg/g ww 8.37 ± 1.62 2.05 ± 1.06 6.49 ± 0.89 6.65 ± 1.83 2.77 ± 0.20 2.35 ± 0.31

Se, adult muscle, μg/g ww 1.50 ± 0.28 0.48 ± 0.15 NT NT NT NT

n

a

297 261 2021 720 1342 209

% fertilization 79.8 ± 4.3 51.5 ± 10.9 81.5 ± 5.0 79.4 ± 5.2 88.0 ± 2.1 94.0 ± 4.8

% mortality 3.3 ± 1.0 0.7 ± 0.4 27.8 ± 7.3 38.3 ±13.7 26.5 ± 4.7 4.2 ± 0.8

% CR 7.7 ± 3.7b0.2 ± 0.2c14.7 ± 3.4 11.7 ± 2.7 10.6 ± 1.9 12.0 ± 4.1

% SK 13.8 ± 5.6b0.7 ± 0.5c19.4 ± 8.2 11.1 ± 2.3 15.6 ± 4.7 4.9 ± 4.9

% FF 3.2 ± 2.0b0.2 ± 0.2c6.8 ± 3.0 15.5 ± 6.6 4.0 ± 0.9 1.5 ± 0.2

% ED 30.8 ± 2 7.4b0.2 ± 0.2c19.9 ± 8.5 13.9 ± 5.3 10.8 ± 2.5 7.5 ± 0.4

% TD 38.9 ± 2 5.6b

0.7 ± 0.5c ND ND ND ND

a

 number of fry to reach the swim-up stage

b and c statistically different values

CR = craniofacial defects, SK = skeletal defects, FF = finfold defects, ED = edema, TD = total defects, NT = not tested, ND = not done
I-18 Draft November 12, 2004

Holm Table 2

Mean embryo-larval parameters for brook trout collected from a high Se site (Luscar Creek), an intermediate Se site (Gregg River), and reference

sites (Cold Creek) in northeastern Alberta over two consecutive years (mean ± SE). Values that are significantly different at " = 0.05 are marked

with different letters. (Table modified from Holm 2002)

Site

2000 2001

Luscar Cold Luscar Gregg Cold

Se, egg, μg/g ww 6.37 ± 0.78 1.35 ± 0.24 8.02 ± 0.77 6.88 ± 0.51 1.25 ± 0.15

Se, adult muscle, μg/g ww 3.79 ± 0.51 0.55 ± 0.10 NT NT NT

n

a

4904 1560 3440 1892 1440

% fertilization 97.4 ± 0.8 96.1 ± 1.2 87.2 ± 2.6 85.2 ± 5.4 77.8 ± 14.2

% mortality 12.6 ± 3.8 9.3 ± 2.4 2.9 ± 0.8 2.9 ± 0.9 3.7 ± 1.6

% CR 13.6 ± 3.5b3.0 ± 0.5c5.6 ± 3.2 2.12 ± 1.0 0.7 ± 0.3

% SK 1.9 ± 0.8 1.3 ± 0.8 2.1 ± 1.1 0.81 ± 0.3 0.6 ± 0.4

% FF 1.1 ± 0.6 1.2 ± 0.8 3.7 ± 1.8 4.1 ± 2.4c0.1 ± 0.1b

% ED 0.6 ± 0.4 0.3 ± 0.1 0.4 ± 0.1 0.3 ± 0.2 1.7 ± 1.2

% TD 14.4 ± 3.6b4.0 ± 2.3c ND ND ND

a

 number of fry to reach the swim-up stage

b and c statistically different values

CR = craniofacial defects, SK = skeletal defects, FF = finfold defects, ED = edema, TD = total defects, ND = not done
I-19 Draft November 12, 2004

Holm Table 3

Mean embryo-larval parameters for rainbow trout and brook trout collected from a high Se site (Luscar

Creek) and reference sites (Deerlick Creek and Cold Creek) in northeastern Alberta over two consecutive

years, combined over both years of the study by site (mean ± SE). Values that are significantly different at

" = 0.05 are marked with different letters. (Table modified from Holm 2002)

Measurement

Rainbow Trout Brook Trout

Luscar Deerlick Luscar Cold

Se, egg, μg/g

ww

6.92 ± 0.78 2.56 ± 0.32 7.20 ± 0.56 1.30 ± 0.14

n

a

2318 1603 8344 3000

% fertilization 81.1 ± 3.9 77.6 ± 5.6 92.3 ± 17.7 88.5 ± 6.2

% mortality 22.2 ± 6.3 19.1 ± 4.6 7.7 ± 2.1 6.9 ± 1.7

% CR 13.1 ± 3.2 7.6 ± 7.1 9.9 ± 2.4 2.7 ± 0.6

% SK 18.1 ± 6.3 11.4 ± 3.8 2.0 ± 0.6 1.0 ± 0.4

% FF 6.0 ± 2.4 2.9 ± 0.8 2.6 ± 1.0 1.2 ± 0.5

% ED 22.4 ± 8.5 7.8 ± 2.2 1.3 ± 0.7 0.9 ± 0.5

a

 number of fry to reach the swim-up stage

CR = craniofacial defects, SK = skeletal defects, FF = finfold defects, ED = edema,

Holm Figure 1. Plot of percent normal (100 - percent edematous) against selenium concentration in adult

rainbow trout muscle ww. 
I-20 Draft November 12, 2004

Holm Figure 2. Plot of percent normal (100 - percent total deformities) against selenium concentration in

adult rainbow trout muscle ww, 2000 data.

Holm Figure 3. Plot of percent normal (100 - percent total deformities) against selenium concentration in

rainbow trout eggs ww, 2001 data. 
I-21 Draft November 12, 2004

Holm Figure 4. Plot of percent normal (100 - percent skeletal deformities) against selenium concentration

in rainbow trout eggs ww, 2001 data. 

Holm Figure 5. Plot of percent normal (100 - percent total deformities) against selenium concentration in

rainbow trout eggs ww, 2001data. EC20 value at 10.4 μg Se/g egg ww.
I-22 Draft November 12, 2004

Brook Trout - 2000 and 2001

Holm Figure 6. Plot of percent normal (100 - total abnormalities) for craniofacial, skeletal and finfold

deformities and edema against selenium concentration in brook trout eggs ww, 2000 and 2001 data.
I-23 Draft November 12, 2004

The effect levels determined using the between streams or within streams approach resulted in values

based on ww in eggs or muscle. Several conversions were necessary to transform a selenium

concentration in egg ww to whole body dw. Using data reported by Holm et al., quantile regression was

used to estimate selenium in adult muscle (ww) from selenium in egg (ww) (see Projection of Muscle

Selenium Concentrations below). A percent moisture of 75.84% derived from rainbow trout data was

used to convert ww to dw and equation 1 was used to convert muscle dw to the whole body dw values

listed below (under Chronic Values).

Projection of Muscle Selenium Concentrations

Median concentrations of selenium in rainbow trout muscles were projected from selenium concentrations

in rainbow trout eggs according to an empirical equation:

[Semuscle] = 0.1827 + 0.1287[Seegg] (R = 0.6244, 5 df)

Parameters of the linear model were estimated by quantile regression, which minimizes the sum of

weighted absolute deviations. Such method is less sensitive to outliers than ordinary least squares

(Koenker and Portnoy 1996). This difference is clearly illustrated in Holm Figure 7: projections of

selenium concentrations in muscles of rainbow trout by the least squares regression line are consistently

greater than projections by the quantile regression line ([Semuscle] = 0.2613 + 0.1418[Seegg]) due to the

disproportional influence of one data point (6.6,1.9). 

Holm Figure 7. Regression lines projecting selenium concentrations in muscles of rainbow trout as a

function of selenium concentrations in rainbow trout eggs.
I-24 Draft November 12, 2004

Chronic Values: Between streams approach

Rainbow trout 2000: effects (craniofacial, skeletal and finfold deformities and

edema) at 1.50 μg Se/g muscle ww or 5.79 μg Se/g dw whole body using

conversion factors listed above; chronic value is 5.79 μg Se/g dw whole body

Brook trout 2000: effects (craniofacial deformities) at 3.79 μg Se/g muscle ww or

13.2 μg Se/g dw whole body using conversion factors listed above; chronic value

is 13.2 μg Se/g dw whole body

Rainbow trout 2001: no effects at 6.65 μg Se/g egg ww or 4.14μg Se/g dw whole

body using conversion factors listed above; chronic value is >4.14 μg Se/g dw

whole body

Brook trout 2001: effects (finfold deformities) at 6.88 μg Se/g egg ww or 12.4 μg

Se/g dw whole body using conversion factors listed above; chronic value is 12.4

μg Se/g dw whole body

Within streams approach

Rainbow trout 2000: no value available; EC20 analysis not appropriate for data

sets

Brook trout 2000: no value available; EC20 analysis not appropriate for data sets

Rainbow trout 2001: EC20 value (craniofacial deformities) at 10.4 μg Se/g egg ww

or 5.85 μg Se/g whole body dw; chronic value is 5.85 μg Se/g whole body dw

Brook trout 2001: no value available; EC20 analysis not appropriate for data set
I-25 Draft November 12, 2004

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: Cutthroat trout (Oncorhynchus clarki lewisi; spawning adults, 3-6 years)

Exposure Route: dietary and waterborne - field exposure

Total selenium concentrations measured at the time the eggs were taken were

<0.1 μg/L from the reference site and 13.3 to 14.5 μg/L at the exposed site.

Study Design: 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 were performed

using 40X magnification.

Effects Data : 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

residue in tissues were as follows:

Site Adult fish liver, 

μg Se/g dw

Adult fish muscle, 

μg Se/g dw

eggs, 

μg Se/g dw

reference 8.2; Range: 3.4-14.6 2.4; 1.4-3.8 4.6

exposed 36.6; Range:18.3-114 12.5; Range: 6.7-41 21.2

Effects >12.5 μg Se/g dw in muscle

Chronic Value: >10.92 μg Se/g dw estimated using the equation I to convert the selenium

concentration in muscle tissue (>12.5 μg Se/g dw) of adult fish to a selenium

concentration in whole-body.
I-26 Draft November 12, 2004

Hardy, R.W. 2002. Effects of dietary selenium on cutthroat trout (Oncorhynchus clarki) growth and

reproductive performance. Annual report for Montgomery Watson Harza. April 30, 2002.

Test Organism: Cutthroat trout (Oncorhynchus clarki, 0.9 g)

Exposure Route: 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 ug/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 ug Se/g dw. Fry were fed initially at

a rate of 10 times per day 6 days a week to apparent saturation. Feeding

frequency decreased as fish grew. 

Test Duration: 124 weeks (865 days, 2.5 yrs)

Study Design: Groups of 50 fish were placed into triplicate tanks (145 L) receiving 4-15 L/min

of hatchery water at 14.5°C 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 of fish 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. 

Effects Data : No signs of toxicity (reduced growth or survival relative to controls) were

observed in fish fed the highest dietary selenium treatment (12 ug Se/g 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. Whole body selenium levels

were approximately 6.8, 10, 12 and 12.5 ug Se/g dw in the four highest dietary

treatments. Nine months later, whole body selenium levels at spawning had

decreased somewhat to 5.21, 8.80, 9.37 and 6.66 ug Se/g dw in these four highest

dietary treatment groups, respectively. Percent survival from the eyed stage to

hatching varied among treatment groups, with the control having the highest

survival (97%) and the fifth dietary treatment group the second highest (93%). 

Percent deformed larvae ranged from a low of 3.4% in controls to a high of 30%

in the 9.0 ug Se/g dw dietary treatment group; larvae in the two highest dietary

treatment groups only exhibited 7 and 6.8 %, respectively. 

Chronic Value: The chronic value for this study is a NOAEC of >9.37 ug Se/g dw whole-body

parent tissue based on embryo/larval deformity. 
I-27 Draft November 12, 2004

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: Fathead minnow (Pimephales promelas; 2 to 8 day-old larvae).

Exposure Route: Dietary only

Green alga, Chlorella pyrenoidosa were exposed to Se (H2

75SeO4

) in culture

water for 3 days. Rotifers, Brachionus calyciflorus, were cultured in chambers

with selenium containing green algae at the ratio of 25 μg algae/ml to 50 μg

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.

Test Duration: 9 to 30 days

Study Design: 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 declorinated 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.
I-28 Draft November 12, 2004

Experiment 1 Experiment 2 Experiment 3

Initial feeding of control diet

(days)

3 8 2

Day Se diet first fed 4 9 3

Day Se diet last fed 11 17 9

Observation days on control diet 19 2 0

Age at study termination (days) 30 19 9

Effects Data:

Experiment 1 Experiment 2 Experiment 3

Mean food Se concentration (μg/g) >70 68 55

Food intake (μg rotifers/larva) 50 1330 1190

Initial larvae mean dry wt. at start

of Se-laden food (μg)

90 400 100

Final larvae mean dry wt. (μg) at

end of test

1470 (Control)

800 (Treatment)a

1888 (Control)

1354 (Treatment)a

475 (Control)

416 (Treatment)

Final mean larval Se content (μg

Se/larva)b

0.0062 0.0700 0.0248

Final mean larval Se

concentrations (μg Se/g dw)

43.0 51.7 61.1

a

 Significantly different from the control.

b

 Values when Se-laden feeding was ended.

Selenium was measured in the test water during the feeding exposures, but the

concentrations were insignificant (0.84 μg/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.

Chronic Value: The estimated whole-body chronic value for this study, determined as the

geometric mean of the final mean larval selenium concentrations measured in the

three experiments, i.e., 43.0, 51.7, and 61.1 μg/g dw, respectively, is 51.40 μg

Se/g dw.
I-29 Draft November 12, 2004

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: Fathead minnows (Pimephales promelas; juvenile, 59 to 61 d old)

Exposure Route: 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.

Test Treatments: 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 μg/g dw. 

Fish used in the first randomized block (F2 generation fish) were progeny from F1

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-blow dried to

5 percent moisture content and crushed and sieved so that only particles retained

by an 11.8 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 μg/L.

Test Duration: 105 days, F2

 generation (block one) and commercial fish (block two);

14 days F3 generation

Study Design: 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.

Effects Data: 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. 
I-30 Draft November 12, 2004

Growth of pre-spawning adults was affected by the selenium exposure. Growth

data are given in the following table:

Effects on Fathead Minnow Growth after 98 days of Exposure to Dietary Selenium

Measured mean selenium in

diet, μg/g dw

Whole-body selenium,

 μg/g dw

Mean fish weight,

 g ww

0.4 1.76 1.30

5.2 2.78 1.24

10.2 3.42 1.20

15.2 5.40 1.21

20.3 6.58 1.09

29.5 7.46 0.94

Chronic Value: An EC20 value could not be calculated for these data because the data did not

meet the minimum requirements for analysis. The MATC for growth of prespawning fathead minnows versus levels of selenium found in whole-body tissue

was the GM of 5.40 and 6.58 μg/g dw, or 5.961 μg Se/g dw.
I-31 Draft November 12, 2004

Schultz, R. and R. Hermanutz. 1990. Transfer of toxic concentrations of selenium from parent to

progeny in the fathead minnow (Pimephales promelas). Bull. Environ. Contam. Toxicol. 45:568-573.

Test Organism: Fathead minnow (Pimephales promelas; Adults)

Exposure Route: Dietary and waterborne

Selenite was added to artificial streams which entered the food web; thus, fish

were also exposed to selenium in the diet. 

Study Design: 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 μg/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 μg/L, respectively. The concentrations of selenium measured in the water

from controls streams were all less than the detection limit, i.e., 2 μg/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 of ten embryo samples were reared in

incubation cups receiving the same streamwater dosed with sodium selenite via a

proportional diluter. The treated embryos in egg cups received an average 9.7 ±

2.6 μg of selenium/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. 

Effects Data : Edema and lordosis occurred in approximately 25 percent of the fish spawned and

reared in 10 μg of selenium/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 μg/g ww. Assuming 85 percent moisture content in the ovaries (see

Gillespie and Baumann below), these concentrations equate to 5.133 and 39.27 μg

Se/g dw.

Chronic Value: <18.21 μg Se/g dw estimated using equation II to convert the selenium

concentration in adult female ovaries (39.27 μg Se/g dw) to a selenium

concentration in whole-body.
I-32 Draft November 12, 2004

Beyers, D.W. and Sodergren, C. 2001a. 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: Larval flannelmouth sucker (Catostomus latipinnis) and larval razorback sucker

(Xyrauchen texanus)

Exposure Route: 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 μg/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 μg/g dw, respectively. Rotifers accumulated selenium from algae

(Chlorella vulgaris) exposed to 0, 25, 50, 100, and 200 μg/L selenate.

Study Design: 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.

Effects Data : 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. 

Chronic Value: The chronic values for the flannelmouth sucker and razorback sucker were >10.2

and >12.9 μg Se/g dw, respectively, based on the concentrations of selenium

measured in whole-body tissue of larval fish at the highest water and dietary

selenium concentrations.
I-33 Draft November 12, 2004

Beyers, D.W. and Sodergren, C. 2001b. 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: Larval razorback sucker (Xyrauchen texanus)

Exposure Route: 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). 

Study Design: 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.

Effects Data : No survival effects were observed. There were no significant decreases in growth

of fish 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 of fish

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 of fish (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 of fish to selenium. 

Chronic Value: 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 μg Se/g dw; Orchard Mesa 

>11 μg Se/g dw; North Pond 50% dilution >41.1 μg Se/g dw; North Pond >42 μg

Se/g dw. Because no significant effects were observed in larvae exposed to North

Pond water at >42 μg Se/g dw whole-body tissue, this value was selected as the

chronic value for the study.
I-34 Draft November 12, 2004

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: Bluegill sunfish (Lepomis macrochirus; embryos and larvae)

Exposure Route: 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 μg/L). A mean selenium

for the ash pond effluent from a previous study was 53 μg/L (N=59; range 35-80

μg/L). 

Study Design: 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 success 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. 

Effects Data : 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 μg/g dw, respectively.

Effect level: < 49, <130 and < 84 μg Se/g dw in adult ovaries, liver and muscle,

respectively

Chronic Value: <59.92 μg Se/g dw estimated using the equation I to convert the selenium

concentration in the muscle of Hyco females (84 μg Se/g dw) to a selenium

concentration in whole-body.
I-35 Draft November 12, 2004

Ingestion Study

Test Organism: Bluegill sunfish (Lepomis macrochirus; 30-day old larvae)

Exposure Route: 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

μg/g dw, respectively. Duration was not given.

Study Design: Survival and observations on pathology and morphology were made in the two

diet treatments.

Effects Data: 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 μg/g dw, respectively.

Effect level for larval survival: <24.7 μg Se/g dw in larvae

Chronic Value: None recommended for larval tissue.
I-36 Draft November 12, 2004

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: Bluegill sunfish (Lepomis macrochirus; embryos and larvae)

Exposure Route: 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.

Study Design: 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 Hycounaffected (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).

Effects Data : 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 μg Se/g dw for adult female

bluegill in ovaries, liver and muscle, respectively
I-37 Draft November 12, 2004

Summary of 28-day embryo larval study

% effluent parent

source in

cross

M X F

% hatch % swimup%

survival,

28-days

adult tissue, μg Se/g dw

gonad liver muscle

M F M F M F

0 H X H 92 0 0 33 30 43 33 62 59

20 H X H 98 0 0 33 30 43 33 62 59

20 H X H 92 0 0 33 30 43 33 62 59

50 H X H 97 0 0 33 30 43 33 62 59

0 H X C 89 87 18 33 2.2 43 4.4 62 2.7

20 H X C 96 96 34 33 2.2 43 4.4 62 2.7

50 H X C 60 84 58 33 2.2 43 4.4 62 2.7

0 C X C 79 95 40 nd 2.2 37 4.4 27 2.7

20 C X C 90 96 36 nd 2.2 37 4.4 27 2.7

20 C X C 88 97 25 nd 2.2 37 4.4 27 2.7

50 C X C 72 92 42 nd 2.2 37 4.4 27 2.7

Chronic Value: <43.70 μg Se/g dw estimated using equation I to convert the selenium

concentration in the muscle of Hyco females (59 μg Se/g dw) to a selenium

concentration in whole-body.

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 μg/g dw; <30 μg/g dw

Adult female liver: >26 μg/g dw, <33 μg/g dw

Adult female muscle: >25 μg/g dw, <59 μg/g dw

Larvae: >12.8 μg/g dw; < 165 μg/g dw
I-38 Draft November 12, 2004

Summary of Embyo/Larval Study up to Swim-up - Affected vs Unaffected Hyco

date of

fert.

Parents’

capture

location in

Hyco

percent hatch percent swim-up selenium in tissue, μg/g dw

at % effluent at % effluent adult female 

0 20 50 0 20 50 ovary liver musc larvae

6-24 affected 93 98 94 0 0 0 30 33 59 0: 130

20: 120

6-27 affected 99 88 77 0 0 0 30 33 59 0: 130

20: 120

6-28 affected 29 34 35 25 14 3 30 33 59 0: 130

20: 120

6-28 affected 98 86 91 5 0 0 30 33 59 0: 130

20: 120

6-29 affected 88 93 85 59 42 25 30 33 59 0: 130

20: 120

7-14 unaffected 92 80 84 79 92 89 9.1 26 25 0: 19

20: 11

50: 10

7-26 unaffected 99 94 93 100 98 98 9.1 26 25 0: 19

20: 11

50: 10

7-27 unaffected 76 84 86 100 89 91 9.1 26 25 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 μg Se/g dw in muscle tissue of Hyco females from the

unaffacted area and >59 μg Se/g dw in muscle tissue of Hyco females from the

affected area. Using equation I to convert the selenium concentration in the

muscle of Hyco females to a selenium concentration in whole-body these values

become >20.29 μg Se/g dw and <43.70 μg Se/g dw, respectively.
I-39 Draft November 12, 2004

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: Bluegill sunfish (Lepomis macrochirus; juvenile- hatchery raised) 

Exposure Route: Dietary only

Test Treatments: 5 diets: Se form (nominal selenium concentration in base diet)

seleno-DL-cystine (5 μg/g)

seleno-DL-cystine (10 μg/g)

seleno-DL-methionine (5 μg/g)

sodium selenite (5 μg/g)

Hyco zooplankton (5 μg/g)

Test Duration: 60 days

Study Design: 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 wholebody.

Effects Data: No decreased length or weight in any of the Se-diets relative to the control.

Chronic Value: all values are whole-body

seleno-DL-cysteine: >2.16 μg Se/g dw

seleno-DL-cysteine-2X: >3.74 μg Se/g dw

seleno-DL-methionine: >2.46 μg Se/g dw

sodium selenite : >1.21 μg Se/g dw

Hyco zooplankton: >2.35 μg Se/g 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 μg Se/g dw for the seleno-DL-cysteine-2X formulation.

Source and Exposure Embryo-Larval Study

Test Organism: Bluegill sunfish (Lepomis macrochirus; Adults from Hyco and a control lake) 

Exposure Route: dietary and waterborne - field exposure

Test Treatments: 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.
I-40 Draft November 12, 2004

Control fish exposed to Hyco water in flow through spawning tanks. 

Hyco-collected fish in control water in flow through spawning tanks.

Test Duration: Adult fish were in spawning tanks 4-7 months 

Study Design: Eggs from each treatment were observed for percent hatch and percent swim-up.

Effects Data: 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

Se in parental

liver tissue,

μg/g dw

water type for

eggs and

larvae N percent hatch

percent swimup

Hyco 18.6 Hyco 16 86.6 91.1

Hyco 18.6 well water 10 83.8 95.5

Control 13.8 Hyco a a 83.3

Control 13.8 well water 12 86.0 97.4

a percent hatch unknown.

Chronic Value: The chronic value for this study is >18.6 μg Se/g dw liver tissue, or >5.45 μg of

Se/g dw whole body tissue using equation III.
I-41 Draft November 12, 2004

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: Bluegill sunfish, wild-caught (Lepomis macrochirus; adults; embryos and larvae)

Exposure Route: dietary and waterborne - field exposure

Test Treatments: 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.

Study Design: 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

recirculated Roxboro City Lake water. Percent hatch (approx. 2d at 22 to 25oC)

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 afer 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).

Effects Data: 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 μg/g wet weight; 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 μg/g wet

weight (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 μg/g wet weight (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 μg Se/g dw.
I-42 Draft November 12, 2004

Chronic Value: <21.47 μg Se/g dw estimated using equation II to convert the selenium

concentration in ovaries of Hyco females (46.30 μg Se/g dw; assuming 85 percent

moisture content) to a selenium concentration in whole-body.
I-43 Draft November 12, 2004

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: Bluegill sunfish (Lepomis macrochirus; two-year old pond-reared adult fish and

resultant fry)

Exposure Route: 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 μg Se/L nominal, 6:1 ratio of selenate:selenite, 98 percent

purity, adjusted to pH 2 with HCl to prevent bacterial growth and change in

oxidation states of Se(IV) and Se(VI).

Test Duration: 140 days

Study Design: 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 μg 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:

Test Treatments

1

(test control)

2

(food control)

3 4 5 6

water 

(μg Se/L)

0.56 8.4 10.5 10.5 10.1 11.0

diet 

(μg Se/g dw)

0.76 0.76 4.63 8.45 16.8 33.3

Effects Data: There was no effect of the combination of highest dietary selenium concentration

(33.3 μg/g dw) in conjunction with exposure to a waterborne selenium

concentration of 11.0 μg/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
I-44 Draft November 12, 2004

exposed to this waterborne and dietary selenium combination was 19 μg/g 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,

μg/g dw

Se in water,

μg/L

whole-body

Se (140 d),

μg/g dw

replicate total no.

spawns

eggs/spawn hatchability,

%

0.8 0.5 0.8 A 15 14,099 94.5

B 10 5,961 90.5

0.8 7.9 1.0 A 12 9,267 89.5

B 11 9,255 84.5

4.6 10.5 3.4 A 20 9,782 86.5

B 12 13,032 96.5

8.4 10.5 6.0 A 2 10,614 96.5

B 9 7,995 90

16.8 10.1 10 A 13 10,797 83

B 13 9,147 91.5

33.3 10.1 19 A 14 8,850 80

B 4 8,850 80
I-45 Draft November 12, 2004

Effects on Larvae

Se in diet, μg/g dw Se in water, μg/L adult whole-body 

(60 d), μg/g dw

mean survival, %

0.8 0.5 0.9 92

0.8 7.9 0.9 93

4.6 10.5 2.9 90

8.4 10.5 4.9 95

16.8 10.1 7.2 87

33.3 10.1 16 7

Chronic Value: The EC20 value calculated for survival of fry versus levels of selenium found in

the eggs and whole-body tissue of adults after 60 d of exposure is 8.954 μg Se/g

dw.
I-46 Draft November 12, 2004
I-47 Draft November 12, 2004

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: Bluegill sunfish (Lepomis macrochirus)

Life Stage: juvenile (5 months - waterborne exposure; 3 months - dietary exposure)

Exposure Route: 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 μg Se/g dw)

Study Design: 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

measured at selected intervals. 

Effects Data : The waterborne exposure (see table below) was determined to have an EC20 =

4.07 μg Se/g dw (1.96-8.44 μg/g 95% CL). However, because it was a water-only

exposure, it was not considered in the derivation of the FCV.

A mortality effect level for the dietary exposure could not be calculated because

the highest selenium whole body concentration (13.4 μg Se/g 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 μg/g dw

(see table below). 

Waterborne Exposure Study

measured selenium in

water (μg/L)

60-d measured

selenium in whole body

(μg/g dw)

60-d mortality (%) condition factor (K)

20 (control) 1.1 10 1.5

160 2.8 12.5 1.5

330 4 22.5 1.6

640 5.3 52.5 1.5

1120 9.8 70 1.6

2800 14.7* 97.5 NA

*A 30-d measurement because all fish were dead at 60 days in this concentration.

Dietary Exposure Study

measured selenium in

food (μg/g ww)

60-d measured

selenium in whole body

(μg/g dw)

60-d mortality (%) condition factor (K)
I-48 Draft November 12, 2004

0.68 (control) 1 5 1.3

2.3 2.1 7.5 1.3

3.5 3.3 10 1.3

6.6 4.7 22.5 1.3

12.7 7.7 15 1.2

25 13.4 17.5 1.2

Chronic Value: Given the very slight reduction in K (1.3 to 1.2) and the uncertain relevance of

growth data, the NOAEC is interpreted to be 13.4 μg Se/g dw for this study; and

the chronic value is >13.4 μg Se/g dw.
I-49 Draft November 12, 2004

Lemly, A.D. 1993a. Metabolic stress during winter increases the toxicity of selenium to fish. Aquatic

Toxicol. 27:133-158.

Test Organism: Bluegill sunfish (Lepomis macrochirus; juvenile 50-70 mm)

Exposure Route: Waterborne and dietary 

Water

1:1 selenite:selenate in stock at pH 2; metered in to reach 5 μg/L 

Diet

seleno-L-methionine in TetraMin (5 μg/g dw)

Test Duration: 180 days

Study Design: 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.

Effects Data : In the 20°C test, fish accumulated 6 μg/g 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 μg/g 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 μg/g dw and lipid levels decrease to 6

percent.

Chronic Value: 20°C, > 6 μg/g Se whole-body; 4°C, < 7.91 μg/g dw Se whole body
I-50 Draft November 12, 2004

Mean Concentration of Selenium in Tissues, Cumulative Survival*, Percent Lipid Content and Oxygen Consumption in Juvenile Bluegill

day

cold - Se control cold + Se warm - Se control warm + Se

 Sea Surv.

%

lipid,

%

O2

b Sea Surv.

%

lipid,

%

O2

b Sea Surv.

%

lipid,

%

O2

b Sea Surv.

%

lipid,

%

O2

b

0 1 100 13.2 98 1 100 13.2 98 1 100 13.2 98 1 100 13.2 98

60 1 97.1 12.5 58 5.8 92.9 10 63 1.2 95.7 13.3 98 5.8 100 13.3 103

120 1.1 97.1 11.5 57 7.9 66.4 6 81 1.1 95.7 13.4 100 6 96.7 13.4 120

180 1.4 97.1 10.5 57 7.9 59.6 6 78 1.2 95.7 13.6 100 6 92.6 13.5 120

a whole body Se tissue concentration, μg/g 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 (μg/g dry weight) of selenium in juvenile bluegill*

day 0 day 60 day 120 day 180

replicate 1 2 3 mean 1 2 3 mean 1 2 3 mean 1 2 3 mean

c+Se 0.87 1.21 0.95 1.01 6.30 5.49 5.76 5.85 8.36 7.31 7.85 7.84 7.53 8.01 8.19 7.91

w+Se 1.17 0.96 0.90 1.01 5.61 6.19 5.43 5.74 6.37 5.92 5.50 5.93 5.48 5.72 6.02 5.74

c-Se 0.89 0.89 0.97 0.97 1.01 1.01 1.10 1.10

w-Se 0.99 0.99 1.12 1.12 0.99 0.99 0.96 0.96

* Each value is for a composite sample made from 5 fish.
I-51 Draft November 12, 2004

The Kaplan-Meier estimator was used to calculate survival at time t

where r(ti

) is the number of fish alive just before time ti

, i.e. the number at risk, and di

 is the number of deaths in the interval Ii

 = [ti

, ti+1]. The 95%

confidence interval for such estimate (Venables and Ripley 2002) was computed as

where

and j # i

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 n.risk n.event survival std.err lower 95% CI upper 95% CI

60 210 15 0.929 0.0178 0.884 0.956

120 165 47 0.664 0.0350 0.590 0.728

180 88 9 0.596 0.0381 0.517 0.666
I-52 Draft November 12, 2004

Hematological Measurements in Juvenile Bluegill Sunfish (*indicates significantly different from control)

Warm Exposure day 0 day 60 day 120 day 180

blood parameter warm-Se warm+Se warm-Se warm+Se warm-Se warm+Se warm-Se warm+Se

total erythrocyte, 106

/ml 2.95 2.92 2.96 2.93 2.99 2.95 2.96 2.89

 % mature 85 86 86 93* 86 94* 85 94*

 nuclear shadows, 104

/ml 0.95 0.86 0.97 2.05* 0.83 2.38* 0.91 2.30*

total leucocytes, 104/ml 17.22 17.41 16.90 17.55 16.73 17.62 17.05 17.36

 % lymphocytes 23 25 20 23 19 26 21 22

 % neutrophils 15 13 14 15 17 19 17 16

hematocrit, % 37 36 37 29* 36 29* 38 28*

MCHC (mean corpuscular hemoglobin conc.) 23 25 25 19* 25 18* 25 17*

Cold Exposure day 0 day 60 day 120 day 180

blood parameter cold-Se cold+Se cold-Se cold+Se cold-Se cold+Se cold-Se cold+Se

total erythrocyte, 106/ml 2.91 2.93 2.97 2.90 3.01 2.95 3.00 2.99

 % mature 84 82 87 95* 85 96* 85 97*

 nuclear shadows, 104

/ml 0.86 0.84 0.83 2.30* 0.89 2.49* 0.90 2.36

total leucocytes, 104/ml 16.48 16.88 16.79 16.91 16.80 16.74 16.96 16.63

 % lymphocytes 17 16 16 17 19 15 19 18

 % neutrophils 13 12 15 11 15 12 12 14

hematocrit, % 39 37 40 30* 41 28* 39 27*

MCHC (mean corpuscular hemoglobin conc.) 26 25 25 18* 22 17* 23 17*

MCV (mean corpuscular volume) 182 171 188 146* 180 135* 185 130*
I-53 Draft November 12, 2004

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.

Test Organism: Bluegill (Lepomis macrochirus; 3 to 4-year old adults)

Exposure Route: 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).

Study Design: Eight Monticello artificial streams were used for three separate studies between

1987 and 1990.

Stream Study I Study II Study III

 Dates

BGa

 put in station 0-2

BG transferred to sta. 6

End of study

9-1-87

5-16-88

8-22-88

10-88

5-89

8-89

11-89

5-90

7-90

1 Unused Control Control 

2 Unused 2.5 μg/L Recovering

3 10 μg/L 10 μg/L Recovering

4 30 μg/L Recovering Recovering

5 Control Control Control

6 30 μg/L Recovering Recovering

7 Control 2.5 μg/L Recovering

8 10 μg/L 10 μg/L Recovering

a

 BG = Bluegill.

A schematic diagram of an artificial stream is provided below. 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.
I-54 Draft November 12, 2004

Schematic Design of One of the Artificial Streams in the Monticello Study

Effects Data : Adult survival in Studies II and III was very low and will not be considered in the

effects analysis. The percent hatch, percent larval survival, percent edema,

percent lordosis and percent hemorrhaging in the 2.5 and 10 μg/L streams for

Study II are provided in the table below. The values presented in this table are

corrected values for Study II as reported by Tao et al. (1999). The data from

Study II (both egg cup and field nest) were not amenable for regression analysis. 

As reported by Tao et al. (1999), ANOVA was utilized to evaluate effects of

elevated concentrations of selenium on percent hatch, percent survival, maximum

percent edema, lordosis, and hemorrhage, and minimum percent healthy (egg cup

data). Treatment effects were only significant for maximum percent edema and

minimum percent healthy (see their Table 4-19), and in no instance were
I-55 Draft November 12, 2004

differences between the 2.5 μg Se/L and control treatments significant (Dunnett’s

Means test, all probabilities > 0.1, see their Table 4-20). These results clearly

suggest that the 2.5 μg Se/L treatment had no adverse impact on bluegill larvae. 

They are further supported by analysis of the field nest data (see table below). In

this experiment, treatment had a significant effect on maximum percent edema

(raw data and ranks) and maximum percent hemorrhage (ranks only). 

Probabilities of differences between the 2.5 μg Se/L and control treatments were

>0.2 for all response variables except maximum percent hemorrhage, which had

an estimated probability of 0.05 (raw data, P=0.022 for ranks; Dunnett’s means

test). Such values, though, were well above the adjusted experiment-wise error

rate for multiple comparison ("’=0.0085, "’=1-(1-")

1/k

; "=0.05, k=6 comparisons;

Sokal and Rohlf 1981), which takes into account the fact that selenium effects

were tested on six different response variables. Therefore, the chronic value for

this study, 12.12 μg Se/g dry weight, was calculated as the geometric mean of

tissue concentrations of selenium in the 2.5 (NOAEC) and 10 μg Se/L (LOAEC)

treatments (5.55 and 26.46 μg Se/g dw whole body tissue, respectively).

Chronic Value: 12.12 μg Se/g dw whole-body tissue, calculated as the GM of the NOAEC, 5.55

μg Se/g dw, and LOAEC, 26.46 μg Se/g dw, based on percent larval survival and

percent larvae exhibiting edema in the egg cup exposures. Note: the NOAEC

value of >17.35 μg Se/g dw was selected as the chronic value for Study III based

on percent larval survival in egg cup exposures and percent larvae exhibiting

edema in nest observations.
I-56 Draft November 12, 2004

Effects on Progeny - Study IIa,b

Egg cup observations

treatment stream

number of

trials % hatch

% survival

to 3rd day % edema % lordosis % hemorr

whole-body Se

(μg/g dw)

control 1 6 93.0 75.2 0 0 0 2.05

control 5 5 96.4 71.5 0 0 0 1.85

2.5 μg/L 2 0 NA NA NA NA NA 6.8

2.5 μg/L 7 4 81.4 71.6 0 0 3.6 5.55

10 μg/L 3 3 83.3 57.7 100 11.1 49.3 20.75

10 μg/L 8 2 91.1 57.1 100 18.2 41.1 33.75

rec 30 μg/L 4 0 NA NA NA NA NA NA

rec 30 μg/L 6 6 92.9 73.0 17.4 0 11.5 30.6

Nest Observations

treatment stream

# active

nests

# embryos

collected

% dead

embryos

# larvae

collected

% dead

larvae

#samples

w larvae % edema % lordosis % hemorr

whole-body Se

(μg/g dw)

control 1 6 2458 0.94 3252 0.03 7 0 0 0 2.05

control 5 9 1329 0 3435 1.05 13 0 0 0 1.85

2.5 μg/L 2 1 0 2497 0.20 3 4.1 25 77.6 6.8

2.5 μg/L 7 5 1462 0 4717 0.08 8 0 0 52 5.55

10 μg/L 3 2 672 0 5376 0.50 9 81.4 5.0 55.5 20.75

10 μg/L 8 3 931 0.32 750 0.40 4 50 14.7 26.7 33.75

rec 30 μg/L 4 0 NA NA NA NA NA NA NA NA NA

rec 30 μg/L 6 8 646 0 6782 7.8 16 27.3 0 17.1 30.6

a Values in table were taken from Tao et al. (1999). 

b The chronic value for the study was calculated as the GM of whole-body selenium concentrations in the 2.5 (NOAEC 5.55 μg Se/g dw;

stream 7 only) and 10 μg Se/L (LOAEC of 26.46 μg Se/g dw; GM of streams 3 and 8, respectively) treatments in the egg cup exposures.
I-57 Draft November 12, 2004

Effects on Progeny - Study IIIa

Egg cup observations

treatment Stream

number of

trials % hatch

% survival to

3rd day % edema % lordosis % hemorr

whole- body

Se (μg/g dw)

control 1 2 92 58.6 0 0 0 1.6

control 5 3 76.7 69.2 0 0.9 0.8 3.35

rec 2.5 μg/L 2 3 87.3 66 0 0 0 5.25

rec 2.5 μg/L 7 6 87.2 76.5 0 0 0 5.35

rec 10 μg/L 3 14.5

rec 10 μg/L 8 3 75.3 74.5 0 0 0 11.7

rec 30 μg/L 4 5 92 78 17.35

rec 30 μg/L 6

Nest observations

treatment stream # active nests

# samples with

larvae % edema % lordosis % hemorr

whole-body Se

(μg/g dw)

control 1 2 5 0 0 0 1.6

control 5 2 3 0 0 0 3.35

rec 2.5 μg/L 2 5 5 0 0 0 5.25

rec 2.5 μg/L 7 5 2 0 0 0 5.35

rec 10 μg/L 3 2 4 0 0 0 14.5

rec 10 μg/L 8 4 4 0 0 0 11.7

rec 30 μg/L 4 9 13 0 0 0 17.35

rec 30 μg/L 6

a The chronic value for the study was selected as the NOAEC of >17.35 μg Se/g dw from the recovering 30 μg Se/L treatment.
I-58 Draft November 12, 2004

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: Striped bass (Morone saxitilis; adults from Lake Norman, NC, approximately 250

g each)

Exposure Route: dietary only

Treated fish were fed selenium contaminated red shiners (1 g) from Belews Lake,

NC (9.6 μg Se/g ww or 38.6 μg Se/g 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 μg Se/g ww or 1.3 μg Se/g dw based on a mean reported moisture

content of 76.3 percent).

Test Treatments: Test treatments were as described above. Two tanks contained treated fish (n =

20 fish total), and one tank of fish 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.

Test Duration: 80 days

Study Design: 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. 

Effects Data: 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 μg 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) μg/g

ww, or 17.5 and 20.0 μg/g dw, respectively, assuming 80 percent moisture

content in muscle tissue. The final selenium concentration in muscle of control

striped bass fed uncontaminated golden shiners averaged 1.1 μg/g ww, or 5.50

μg/g dw (assuming 80 percent moisture content in muscle tissue).
I-59 Draft November 12, 2004

Chronic Value: The chronic value for percent survival of striped bass relative to final selenium in

muscle tissue after being fed Se-laden red shiners is <17.50 μg/g dw, or 14.75

μg/g dw whole body tissue converted using equation I.

An EC20 value could not be calculated for this data set because the data did not

meet the assumptions required for analysis.
I-60 Draft November 12, 2004

Lemly, A.D. 1993b. Teratogenic effects of selenium in natural populations of freshwater fish. 

Ecotoxicol. Environ. Safety. 26: 181-204.

Test Organism: All possible fish species collected from Belews Lake and a reference site.

Exposure Route: dietary and waterborne - field exposed

Study Design: Surveys of external abnormalities in fish collected from Belews Lake and two

reference lakes were done in 1975, 1978, 1982, and 1992. Five classifications of

abnormalities were reported: (1) spinal deformities (lordosis, scoliosis, kyphosis);

(2) accumulation of body fluid (edema, expothalmus or popeye); (3) missing or

abnormal fins; (4) abnormally shaped head or mouth; and (5) cloudy eye lens or

cornea (cataracts). Whole-body selenium was measured in each fish. The

relationship between whole-body selenium and malformations was examined.

Effects Data : The relationship between whole-body selenium and the frequency of

malformations in all the fish species collected at Belews (n=22) did not follow a

clear trend. When evaluating only fish from the family Centrarchidae using a

polynomial regression (cubic model) an R2

 value of 0.881 was obtained. Lemly

reported that the inflection point where a rapid rise in deformities occurred was

between 40 and 50 μg Se/g dw in whole-body tissue. The EC20 value determined

by regression analysis of percent normal fish versus whole-body tissue selenium

concentration for the family Centrarchidae (most sensitive family or group of

families) was 44.57 μg Se/g dw. Centrarchidae was the most sensitive family or

group of families of those collected during the survey.

Chronic Value: The EC20 value determined by regression analysis of percent normal fish versus

whole-body tissue selenium concentration for the family Centrarchidae was 44.57

μg Se/g dw.
I-61 Draft November 12, 2004
J-1 Draft November 12, 2004

APPENDIX J

SELENIUM (μg/g dw WHOLE-BODY) IN FISH SAMPLES COLLECTED FROM 112

SITES AS PART OF U.S. FISH AND WILDLIFE’S NATIONAL BIOMONITORING

PROGRAM, 1978-1981 (LOWE ET AL. 1985).

AND

SELENIUM (μg/g dw WHOLE-BODY) IN 322 AQUATIC LIFE TISSUE SAMPLES

COLLECTED FROM 264 SITES AS PART OF USGS NATIONAL WATER QUALITY

ASSESSMENT (NAWQA) PROGRAM

(http://water.usgs.gov/nawqa/ as of May 11, 2004).
J-2 Draft November 12, 2004

FCV Relative to Natural Background Levels of Selenium in Fish

As an essential element, selenium naturally occurs in all living things. Since selenium is found in all fish,

two questions arise. 1) How close is the FCV of 7.91 μg/g dw to natural background levels in fish, and 2)

how frequently do natural selenium tissue concentrations exceed the FCV. The latter situation would pose

problems in the implementation of the FCV as an ambient water quality criterion.

As part of the National Contaminant Biomonitoring Program, the U.S. Fish and Wildlife Service collected

fish from 112 sites distributed evenly across the U.S. during 1979 through 1981, and measured several

contaminants including selenium (Lowe et al. 1985). Selenium, measured in 591 fish samples representing

60 different species, ranged from 0.3 to 10.5 μg/g dw and had an overall average and standard deviation of

1.9 ± 1.4 μg/g dw. 

A separate data set of selenium levels in 231 macroinvertebrate samples, 90 fish samples, and one plant

sample collected from 25 different states across the United States was generated by USGS’s National

Water Quality Assessment (NAWQA) program. NAWQA is intended to measure water quality in a

sampling of smaller watersheds having known land use. Among these sites, whole body tissue

concentrations ranged from 0.3 to 22.37 μg/g dw and had an overall average and standard deviation of 3.22 

± 2.29 μg/g dw. The distribution of both these data sets indicates that the FCV would not be exceeded by

over 97 percent of aquatic tissue samples collected across the United States (Figure J-1). The FCV thus

appears to be sufficiently greater than natural selenium levels that unavoidable exceedances of the criterion

are unlikely.
J-3 Draft November 12, 2004

Distribution of selenium concentrations in whole body 

0.0

0.2

0.4

0.6

0.8

1.0

0 5 10 15 20 25

Se in whole body [μg/g dw]

Cumulative proportion

National Contaminant Biomonit. Pgm (fish

samples) n = 591

USGS NAWQA Sites (fish and invertebrate

samples) n = 322, 64 nondetects (19.9%)

Chronic criterion 

7.9 μg/g dw

0.986

0.967

Figure J-1. Cumulative distribution of selenium concentrations in aquatic organisms (whole-body, μg/g

dw) collected by the National Contaminant Biomonitoring Program (NCBP) and the U.S.

Geological Service National Water-Quality Assessment (NAWQA) Program. NCBP and

NAWQA data from Lowe et al. (1985) and query results from NAWQA’s database on

contaminant concentrations in animal tissues (http://water.usgs.gov/nawqa/), respectively.
J-4 Draft November 12, 2004

Table J-1. Selenium (μg/g dw whole-body) in fish samples collected from 112 sites as part of U.S. Fish and

Wildlife’s National Biomonitoring Program, 1978-1981. From Lowe et al. 1985

Year Species Mean total length, cm Mean total weight, Kg Se, ug/g dry wt.

Station 1, Penobscot River at Old Town, MA

78 Smallmouth bass 12.9 1.2 0.8562

78 White sucker 13.7 1.1 1.2227

78 White sucker 14.4 1.3 0.9292

80 Smallmouth bass 12.8 1.1 0.6513

80 White sucker 15.2 1.3 0.8261

80 White sucker 15.3 1.4 0.7634

Station 2, Connecticut River at Windsor Locks, Conn.

78 White catfish 16.6 2.3 0.4651

78 White catfish 16.5 2.3 0.6818

78 Yellow perch 8 0.3 0.9934

80 White catfish 14.5 0.9 0.6007

80 White catfish 13.3 0.9 0.9738

80 Yellow perch 9.5 0.4 0.9811

Station 3, Hudson River at Poughkeepsie, NY

78 Goldfish 11 1 0.9353

78 Goldfish 11.4 1.1 0.6545

78 Largemouth bass 11.1 0.8 1.0676

80 Goldfish 10.9 1 1.2333

80 Largemouth bass 14.8 2.2 1.0701

Station 4, Delaware River at Trenton, NY- Yardley, Pa.

79 White perch 7.3 0.2 4.6429

79 White sucker 12.8 0.8 1.1438

79 White sucker 14.3 1.2 0.8389

81 Largemouth bass 9.5 0.4 2.4206

81 White sucker 15 1.3 1.1864

81 White sucker 14.4 1.1 1.4423

Station 5, Susquehanna River at Conowingo Dam, Md.

79 Common carp 12.9 2 2.0690

79 Common carp 16.9 2.3 2.2381

79 White perch 7.6 0.3 5.5401

81 Common carp 14.4 1.6 2.5431

81 Common carp 14.1 1.7 1.5358

81 White perch 7.9 0.3 3.4951

Station 6, Potomac River at Little Falls Md.- McLean Va

79 Common carp 18.7 3.1 1.5248

79 Common carp 17 2.5 1.1628

79 Smallmouth bass 10 0.5 2.6587

81 Largemouth bass 11.5 8 1.8474

81 Redhorse 17.2 2 1.2963

81 Redhorse 17.5 2.1 1.3208

Station 7, Roanoke river at Roanoke Rapids, N.C.
Year Species Mean total length, cm Mean total weight, Kg Se, ug/g dry wt.

J-5 Draft November 12, 2004

78 White catfish 12.7 0.7 1.2134

80 Striped bass 14.5 1.4 1.3665

80 White catfish 11.7 0.6 1.0473

80 White catfish 10.1 0.4 1.0164

Station 8, Cape Fear River at Elizabethtown, NC

78 Spotted sucker 16.3 2 2.5177

78 Spotted sucker 16 2 2.5263

80 Flathead catfish 19 2 1.0656

80 Quillback 15 1.7 1.6719

80 Quillback 14.9 1.1 1.6558

Station 9, Cooper River at Lake Moultrie, Monck;s Coner, S.C

78 Channel catfish 16.3 1.3 1.6078

78 Channel catfish 14.6 1 1.4563

80 Channel catfish 14.5 1 1.4497

80 Channel catfish 13.6 0.6 1.4917

80 Striped bass 20.6 3.3 1.4894

Station 10, Savannah River at Savannah, Ga

78 Channel catfish 11 0.4 3.2444

78 White catfish 12.7 1 2.0248

80 White catfish 11.3 0.7 1.4592

80 White catfish 7.9 0.2 1.2319

80 Bowfin 21 3.6 2.2568

Station 12, St. Lucie Canal at Indiantowm, Fla

78 Largemouth bass 1.0954

78 White catfish 1.0580

78 White catfish 0.7931

80 Largemouth bass 1.1837

80 White catfish 1.3208

80 White catfish 0.9690

Satation 13, Appalachicola River at J. Woodruff Dam, Fla.

79 Largemouth bass 0.8803

79 Spotted sucker 1.8219

79 Spotted sucker 1.0980

81 Largemouth bass 0.9402

81 Spotted sucker 1.3060

81 Spotted sucker 1.5600

Station 14, Tombigbee Tiver at McIntosh, Ala.

79 Smallmouth buffalo 0.7325

79 Smallmouth buffalo 1.1513

81 Black crappie 1.2545

81 Blue catfish 0.8765

81 Blue catfish 0.7782

Station 15, Mississippi Tiver at Luling, La.
Year Species Mean total length, cm Mean total weight, Kg Se, ug/g dry wt.

J-6 Draft November 12, 2004

79 Common carp 1.6667

79 Common carp 1.7162

79 Largemouth bass 1.7200

81 Channel catfish 0.6599

81 Channel catfish 0.7561

81 Largemouth bass 1.8147

Station 16, Rio Grande at Mission, Tex

80 Gizzard shad 2.4638

80 Gizzard shad 2.4719

80 Largemouth bass 2.2800

81 Common carp 2.1858

81 Gizzard shad 2.6190

81 Gizzard shad 2.8125

Station 17, Genessee River at Scottsville, NY

80 Pumpkinseed 0.9901

80 Redhorse 0.7692

80 Redhorse 0.7328

81 Pumpkinseed 2.1186

81 Redhorse 1.2450

81 Redhorse 1.3853

Station 18,, Lake Ontario at Prot Ontario, NY

78 Rock bass 1.1355

79 Yellow perch 1.3306

79 Yellow perch 1.1719

81 Rock bass 1.4886

81 Yellow perch 1.7293

81 Yellow perch 1.3383

Station 19, Lake Erie at Erie, Pa

80 Redhorse 1.7625

80 Redhorse 1.7241

80 Yellow perch 2.4576

Station 20, Lake Huron (Saginaw Bay) at Bay port, Mich.

79 Common carp 1.8237

79 Common carp 1.9113

79 Yellow perch 1.9196

81 Common carp 2.3355

81 Common carp 2.5776

81 Yellow perch 2.1723

Station 21, Lake Michigan at Sheboygan, Wis.

79 Bloater 0.8060

79 Bloater 0.6897

79 Lake trout 1.1730

81 Bloater 0.7104
Year Species Mean total length, cm Mean total weight, Kg Se, ug/g dry wt.

J-7 Draft November 12, 2004

81 Bloater 0.9687

81 Lake trout 1.2828

Station 22, Lake Superior at Bayfield, Wis.

79 Lake trout 0.8911

79 Lake whitefish 1.3278

79 Lake whitefish 1.5058

81 Bloater 1.1304

81 Bloater 1.3419

81 Lake trout 1.4741

Station 23, Kanawha River at Winfield, W.VA

78 Channel catfish 0.9091

78 Channel catfish 0.8841

78 Sauger 1.4334

80 Channel catfish 0.9508

80 Channel catfish 1.2635

80 Sauger 2.3651

Station 24, Ohio River at marietta, Ohio- Williamstown, W VA

78 Channel catfish 1.1871

78 Sauger 1.4716

80 Common carp 2.2819

80 Common carp 1.7687

80 Sauger 2.2511

Station 25, Cumberland River at Clarksville, Tenn.

78 Common carp 1.3793

78 common carp 1.8077

78 White catfish 1.2203

80 Common carp 1.3514

80 Common carp 1.5909

80 Largemouth bass 1.7669

Station 26, Illinois River at Beardstown, Ill.

78 Black crappie 0.8638

78 Common carp 2.0438

78 Common carp 2.6766

80 Black crappie 1.5751

80 Common carp 1.8051

80 Common carp 2.1687

Station 27, Mississippi River at Gutenburg, Iowa- Glen Haven, Wis.

78 Common carp 1.7628

78 Common carp 1.3907

78 Largemouth bass 2.2742

80 Common carp 1.3231

80 Common carp 0.9064

80 Largemouth bass 1.1885
Year Species Mean total length, cm Mean total weight, Kg Se, ug/g dry wt.

J-8 Draft November 12, 2004

Station 28, Arkansas River at Pine Bluff, Ark.

79 Bluegill 0.8936

79 Common carp 1.2453

79 Common carp 1.0357

81 Common carp 1.7931

81 Common carp 1.3937

81 Largemouth bass 1.3011

Station 29, Arkansas River at Keystone Reservoir, Okla.

79 Common carp 1.3974

79 Common carp 1.6509

79 White bass 2.2167

81 Common carp 2.2394

81 Common carp 1.3410

81 White crappie 1.0738

Station 30, White River at De Valls Bluff, Ark.

79 Freshwater drum 0.8874

79 Freshwater drum 0.9091

79 Largemouth bass 0.8696

81 Common carp 2.2857

81 Common carp 1.7472

Station 31, Missouri River at Nebraska City, Nebr.- Hamburg, Iowa

79 Common carp 1.8774

79 Common carp 2.8163

79 Goldeye 1.2712

81 Common carp 3.0189

81 Common carp 3.2051

81 Goldeye 3.1803

Station 32, Missouri River at Garrison Dam, N. Dak.

79 Northern pike 1.4884

79 Redhorse 0.9600

81 Walleye 1.6041

81 White sucker 2.4883

81 White sucker 3.9252

Station 33, Missouri River at Great Falls Mont.

79 Brown trout 2.3432

79 White sucker 1.3333

79 White sucker 1.3158

81 Brown trout 2.1591

81 White sucker 1.9617

Station 34, Red River of the Norh at Noyes, Minn. _ Pembina, N. Dak.

78 Common carp 2.3166

78 Common carp 2.0629

78 Sauger 0.4682

80 Mooneye 3.3754
Year Species Mean total length, cm Mean total weight, Kg Se, ug/g dry wt.

J-9 Draft November 12, 2004

80 Sauger 0.9328

80 Sauger 0.8117

Station 35, Green River at Vernal, Utah

78 Common carp 3.7410

78 Common carp 3.9286

78 Smallmouth bass 3.6076

80 Common carp 3.2537

80 Common carp 2.7811

80 Smallmouth bass 3.1500

Station 36, Colorado River at Imperial Reservoir, Ariz.- Calif.

78 Common carp 6.5552

78 Common carp 8.0364

78 Largemouth bass 10.5204

80 Common carp 7.5210

80 Common carp 6.4783

80 Largemouth bass 8.6531

Station 37, Truckee River at Fernley, Nev.

80 Green sunfish 1.0794

80 Tahoe sucker 0.9211

80 Tahoe sucker 1.1401

81 Green sunfish 0.8835

Station 38, Utah lake at Provo, Utah

78 Common carp 2.9333

78 Common carp 3.1741

78 White bass 3.4799

80 Common carp 9.6863

80 Common carp 2.1633

80 White bass 3.5246

Station 39, Sacramento River at Sacramento, Calif.

79 Brown bullhead 0.7035

79 Largemouth bass 1.2644

79 largescale sucker 1.0811

81 largescale sucker 1.2454

81 Largemouth bass 1.4286

Station 40, San Joaquin River at Los Banos, Calif.

79 Black bullhead 3.3333

79 Black bullhead 3.3871

79 Green sunfish 6.0748

81 Sacramento blackfish 5.3425

81 Sacramento blackfish 5.7407

Station 41, Snake River at Hagerman, Idaho

78 Largescale sucker 1.2431

78 Largescale sucker 1.4126
Year Species Mean total length, cm Mean total weight, Kg Se, ug/g dry wt.

J-10 Draft November 12, 2004

78 Rainbow trout 2.3630

80 Largescale sucker 1.3913

80 Largescale sucker 1.5447

80 Rainbow trout 1.9495

Station 42, Snake River at Lewiston, Idaho- Clarkston, Wash.

78 Largescale sucker 0.9325

78 Largescale sucker 1.3636

78 Smallmouth bass 1.4765

80 Largescale sucker 0.8861

80 Largescale sucker 0.9746

80 White crappie 1.0870

Station 43, Salmon River at Riggins, Idaho

78 Bridgelip sucker 1.5719

78 Bridgelip sucker 0.8494

78 Northern squawfish 1.1930

80 Bridgelip sucker 0.9016

80 Bridgelip sucker 0.8475

80 Northern squawfish 2.9897

Station 44, Yakima River at Granger, Wash.

78 Common carp 2.3026

78 Common carp 1.4047

80 Black crappie 1.6716

80 Largescale sucker 1.7742

80 Largescale sucker 1.6508

Station 45, Willamette River at Oregon City, Oreg

78 Northern squawfish 0.5078

78 Chiselmouth 0.6615

78 Chiselmouth 0.4082

80 Largescale sucker 0.5479

80 Largescale sucker 0.6907

80 Northern squawfish 1.4286

Station 46, Columbia River at Cascade Locks, Wash. -Oreg.

78 Largescale sucker 1.2684

78 Largescale sucker 1.3712

78 Northern squawfish 1.7818

80 Largescale sucker 0.9236

80 Largescale sucker 0.6765

80 Northern squawfish 0.7025

Station 47, Klamath River at br k, Calif. 

79 Klamath largescale sucker 0.3409

79 Klamath largescale sucker 0.3774

79 Yellow perch 0.6693

81 Klamath largescale sucker 1.0121
Year Species Mean total length, cm Mean total weight, Kg Se, ug/g dry wt.

J-11 Draft November 12, 2004

81 Yellow perch 0.9016

Station 48, Rogue River at G )Id, y Da Oreg. 

79 Black crappie 0.3836

79 Redside shine 0.4887

81 Black crappie 0.3158

81 Brown bullhead 0.7805

81 Brown bullhead 0.7692

Station 49, (,hena River at rks, aska 

79 Burbot 2.3005

79 Longnose sucker 1.2903

81 Longnose sucker 1.7757

81 Longnose sucker 1.8519

81 Northern pike 1.8026

Station 50, Kenai River at SDidatna, laska

78 Rainbow trout 2.0391

78 Round whitefish 2.9538

78 Dolly Varden 1.6992

80 Rainbow trout 1.8910

80 Round whitefish 1.8954

80 Dolly Varden 1.6910

Station 51, Kennebec Rive at iiic y, Maine 

78 White sucker 1.1060

78 White sucker 0.9692

78 Yellow perch 1.2549

80 White sucker 1.0046

80 White sucker 0.9459

80 Yellow perch 0.7011

Station 52 Lake Champlain Burlic gton, Vt.

78 Northern pike 0.7451

78 White sucker 0.8400

78 White sucker 0.8676

80 Northern pike 1.1163

Station 53, Menimack River t Low@ll, Mass. 

78 Largemouth bass 0.8070

78 White sucker 1.0357

78 White sucker 1.0676

80 Smallmouth bass 0.7343

80 White sucker 0.8230

80 White sucker 1.2389

Station 54, Raritan River at Highland Park, N.J.

78 Largemouth bass 1.8060

78 White sucker 1.9454
Year Species Mean total length, cm Mean total weight, Kg Se, ug/g dry wt.

J-12 Draft November 12, 2004

78 White sucker 1.8301

80 White sucker 1.5126

80 White sucker 2.3348

80 Redfin pickerel 1.6450

Station 55, James River at Richmond, Va. 

79 Redhorse 1.2877

79 Redhorse 1.4194

79 Smallnouth bass 1.8657

81 Redhorse 1.9243

81 Redhorse 1.0658

81 Smallnouth bass 1.0359

Station 56, Pee Dee River at Johnsonville, S.C. 

80 Gizzard shad 1.1170

80 Gizzard shad 1.0000

80 Largemouth bass 1.5235

Station 57, Altamaha River at Doctortown, Ga. 

78 Black crappie 1.2857

78 Carpsucker 1.2342

80 Largemouth bass 1.7094

80 Spotted sucker 2.0408

80 Spotted sucker 1.5574

Station 59, Alabama Rive'r at Chrysler, Ala.

79 Smallmouth buffalo 1.0035

79 Smallmouth buffalo 1.0175

79 Bowfin 1.2203

81 Largemouth bass 1.1538

81 Blue catfish 0.6716

81 Blue catfish 0.7326

Station 60, Brazos River a@ Richmond, Tex.

79 Longnose gar 1.2681

79 Smallmouth buffalo 1.0320

79 Smallmouth buffalo 1.3693

Station 61, Colorado River at Wharton, Tex. 

79 Channel catfish 0.9662

79 Freshwater drum 1.7844

79 Freshwater drum 1.4943

Station 63, Rio Grande at Elephant Butte, N. Mex.

78 Common carp 2.1514

78 Common carp 1.9028

78 Largemouth bass 1.9310

80 Common carp 1.7597

80 Common carp 1.5830
Year Species Mean total length, cm Mean total weight, Kg Se, ug/g dry wt.

J-13 Draft November 12, 2004

80 Largemouth bass 1.5709

Station 64, Rio Grande at Alamosa, Colo.

78 White sucker 0.7442

78 White sucker 0.9231

80 Brown trout 1.2775

80 White sucker 0.6911

80 White sucker 0.8036

Station 65, Pecos River at Red Bltiff Lake, Tex. 

78 Gizzard shad 4.2715

78 White bass 9.5016

80 Gizzard shad 3.8559

80 Gizzard shad 5.0673

80 White bass 6.0681

Station 66, St. Lawrence River at Massena, N.Y.

79 Smallmouth bass 1.1765

79 White sucker 1.0280

79 White sucker 1.4414

81 Northern pike 1.3592

Station 67, Allegheny River at Natrona, Pa.

78 Redhorse 2.0155

78 Redhorse 1.4232

78 Smallmouth bass 2.2794

79 Largemouth bass 1.3693

79 Redhorse 1.9005

79 Redhorse 1.5789

80 Redhorse 2.7511

80 Redhorse 2.8139

80 Smallmouth bass 2.2656

Station 68, Wabash River at New Harmony, -Crossville, III

78 Common carp 2.0505

78 Common carp 2.2302

78 Largemouth bass 2.3413

80 Common carp 1.3043

80 Common carp 1.4873

80 Largemouth bass 1.5175

Station 69, Ohio River at Cincinnati, Ohio

78 Common carp 2.5890

78 Common carp 4.0333

78 Sauger 1.5031

80 Common carp 2.1071

80 Common carp 2.6070

80 Sauger 1.5113
Year Species Mean total length, cm Mean total weight, Kg Se, ug/g dry wt.

J-14 Draft November 12, 2004

Station 70, Ohio River at Metropolis Ill.-Paducah, Ky.

78 Common carp 2.2222

78 Common carp 1.4067

78 Largemouth bass 1.3514

80 Common carp 2.2118

80 Common carp 2.0599

80 Largemouth bass 1.7770

Station 71, Tennessee iver at Savannah, Tenn.

78 White bass 1.4610

79 Common carp 2.3790

79 Carpsucker 1.3333

80 Common carp 2.1973

80 Common carp 1.8103

80 Largemouth bass 1.9178

Station 72, Wisconsin River at Woodman, Wis.

80 Common carp 1.4107

80 Common carp 1.1688

80 Largemouth bass 1.1538

Station 73, Des Moines River at Keosauqua, Iowa

78 Common carp 3.5986

78 Common carp 4.0956

78 Sauger 2.4706

80 Common carp 3.8462

80 Common carp 2.3221

80 Channel catfish 1.7883

Station 74, Mississippi River at Little Falls, Minn.

78 White sucker 1.4340

78 Yellow perch 1.5825

80 rock bass 1.6207

80 Yellow bullhead 2.0155

80 Yellow bullhead 1.9149

Station 75, Mississippi River at Cape Girardeau, Mo.-Ill

78 Common carp 2.3511

78 Common carp 2.1019

78 White crappie 1.2360

80 Common carp 1.4449

80 Common carp 1.8077

80 White bass 2.8839

Station 76, Mississippi River 

79 Bluegill 1.8000

79 Smallmouth buffalo 0.6818

79 Smallmouth buffalo 0.6140

81 Smallmouth buffalo 1.0979

81 Smallmouth buffalo 0.9884
Year Species Mean total length, cm Mean total weight, Kg Se, ug/g dry wt.

J-15 Draft November 12, 2004

81 White crappie 0.9343

Station 78, Verdigris River at Oologah Okla.

79 Common carp 1.0268

79 Common carp 1.4218

79 White bass 1.9907

81 Bluegill 1.7063

81 Common carp 2.1223

81 Common carp 2.1456

Station 79, Canadian River at Eufaula, Okl.

79 Common carp 2.4324

79 Common carp 1.4113

79 Largemouth bass 1.6318

81 Common carp 1.8382

81 Common carp 1.9403

81 Largemouth bass 2.3789

Station 80, Yazoo River at Redwood, Miss

79 Common carp 1.1620

79 Common carp 2.7356

81 Smallmouth buffalo 1.1875

81 Smallmouth buffalo 1.3559

81 White crappie 1.4444

Station 81, Red River at Alexandria.

79 Smallmouth buffalo 0.8929

79 Smallmouth buffalo 0.9667

79 White bass 1.3043

81 Freshwater drum 1.4103

81 Freshwater drum 1.2500

81 Spotted gar 0.7353

Station 82, Red River at Lake Texoma, Okla.-Tex.

79 Black crappie 1.2955

79 River carpsucker 1.3366

79 River carpsucker 1.7617

81 Common carp 1.8280

81 Common carp 2.4627

81 Largemouth bass 2.3043

Station 83, Missoun River at Hermann, Mo.

79 River carpsucker 0.9121

79 River carpsucker 1.1350

79 Smallmouth buffalo 1.4706

Station 84, Bighorn River at Hardin, Mont. 

79 Common carp 5.6522

79 Goldeye 9.4118

79 White sucker 6.9466
Year Species Mean total length, cm Mean total weight, Kg Se, ug/g dry wt.

J-16 Draft November 12, 2004

81 Brown trout 5.0896

81 Longnose sucker 3.0717

Station 85, Yellowstone River at Sidney,Mont. 

79 Common carp 1.4773

79 Common carp 1.9277

79 Sauger 1.7257

81 Redhorse 1.8919

81 Redhorse 2.4832

81 Sauger 1.6832

Station 86, James River at Olivet, S. Dak vet, S. ak. 

79 Carpsucker 1.7188

79 Carpsucker 1.9184

79 Goldeye 1.7302

81 Carpsucker 1.0154

81 Carpsucker 1.2805

81 Goldeye 2.5185

Station 87, North Platte River at Lak McConaughy, Nebr. 

79 Common carp 3.7288

79 Common carp 4.8918

79 Walleye 1.4907

81 Common carp 3.0488

81 Common carp 2.6601

81 Walleye 2.0077

79 Black crappie 2.7881

79 Common carp 4.3902

79 Common carp 4.3590

Station 88, South Platte River at Brule, Nebr. 

81 White sucker 4.6538

81 Orangespotted sunfish 8.6786

Station 89, Platte River at LduisviIle Nebr.

79 Carpsucker 2.2549

79 Carpsucker 1.6514

79 Goldeye 3.2335

81 Carpsucker 2.5207

81 Carpsucker 2.8270

81 Goldeye 4.0972

Station 90, Kansas River at onner prings, Kans. 

79 Common carp 2.4615

79 River carpsucker 1.0676

81 Common carp 1.5858

81 Channel catfish 2.1635

81 River carpsucker 0.9859

Station 91 Colorado River at Lake Havasu, Ariz.-Calif.
Year Species Mean total length, cm Mean total weight, Kg Se, ug/g dry wt.

J-17 Draft November 12, 2004

78 Common carp 7.6490

78 Largemouth bass 5.1449

80 Common carp 3.6494

80 Common carp 5.6944

80 Largemouth bass 2.7666

Station 92, Colorado River at Lake Mead Ariz.-Nev. 

79 Channel catfish 4.6441

79 Channel catfish 3.0169

79 Striped bass 0.8182

81 Common carp 3.3735

81 Channel catfish 3.2707

81 Striped bass 3.7109

Station 93, Colorado v" t Lake 11, Ariz. 

78 Common carp 9.4218

78 Largemouth bass 9.8990

80 Common carp 4.3922

80 Common carp 3.9574

80 Largemouth bass 2.3759

Station 94, Gfla River at Sa "IIDI servoir, Ariz. 

78 Common carp 1.9231

78 Common carp 1.5918

78 Largemouth bass 1.7466

80 Common carp 1.9588

80 Common carp 1.4079

80 Largemouth bass 1.1524

Station 96, Snake River at I H @b, 'D.@'oWash

78 Largescale sucker 1.0000

78 Largescale sucker 1.2625

78 Northern squawfish 0.8970

80 Largescale sucker 0.8765

80 Largescale sucker 0.8456

80 Northern squawfish 1.7316

Station 97, Columbia River Pa,,O, sh.

78 Yellow perch 3.8667

78 Chiselmouth 1.6242

78 Chiselmouth 1.2375

80 Common carp 4.0244

80 Common carp 2.6923

80 Yellow perch 3.5662

Station 98, Columbia i,er G @,n 'C,uloee, Wash.

78 Largescale sucker 0.8300

78 Largescale sucker 0.9266

78 Yellow perch 1.1847

80 Largescale sucker 0.7692
Year Species Mean total length, cm Mean total weight, Kg Se, ug/g dry wt.

J-18 Draft November 12, 2004

80 Largescale sucker 0.7519

80 Walleye 0.8108

tation 99, Waikele Stea, Waipah Hawaii 

79 Cuban limia 4.0755

Station 100, Manoa Stream at Honol Hawai

79 Cuban limia 3.5577

79 Mazambique tilapia 1.6502

Station 101, Androscoggin iver at wiston, Main

78 White sucker 0.9426

78 White sucker 0.7059

78 Yellow perch 0.7042

80 White sucker 0.6299

80 White sucker 0.5691

80 Yellow perch 0.8961

Station 102, Laice Superior at@Keewe@naw Point, Mich. 

79 Bloater 1.2798

79 Bloater 0.9385

79 Lake trout 0.7339

81 Bloater 1.3354

81 Bloater 1.6961

81 Lake trout 1.4286

Station 103, Lake Superior at Whitefish Point, Mich.

79 Lake trout 0.7427

79 Lake whitefish 1.1379

79 Lake whitefish 1.8051

81 Lake trout 1.0617

81 Lake whitefish 1.4947

104, Lake Michigan at Beavei,lsland,Mich.

79 Bloater 0.8537

79 Bloater 0.4963

79 Lake trout 1.3483

Station 105. Lake Michigan at Saugat@ck, Mich

79 Bloater 0.4948

79 Bloater 0.6651

79 Lake trout 0.8310

81 Bloater 0.8696

81 Bloater 0.8939

81 Lake trout 1.1480

Station 106, Lake Huron at Alpena Mich. 

79 White sucker 1.6596

79 White sucker 1.3793

79 Yellow perch 2.5556
Year Species Mean total length, cm Mean total weight, Kg Se, ug/g dry wt.

J-19 Draft November 12, 2004

81 Lake trout 1.2808

81 White sucker 2.3293

81 White sucker 2.2403

Station 107, Lake St. Clair at Mount Clements. Mich.

79 Common carp 1.2030

79 Common carp 1.6418

79 Walleye 1.7731

81 Common carp 1.8182

81 Common carp 2.0861

81 Walleye 1.9101

Station 108, Lake Erie at Port Clinton. Ohio

79 Common carp 1.4696

79 Common carp 1.4483

79 Walleye 0.6329

81 Common carp 2.1951

81 Common carp 1.6992

81 Walleye 1.0811

Station 109, Lake Ontario at Roosevelt Beach, N.Y.

79 Brown trout 1.1184

79 Rock bass 1.7626

79 Rock bass 1.4179

81 Lake trout 1.1310

81 Rock bass 1.6988

81 Rock bass 2.0949

 Station I I 1. Mississippi River at Lake City, Minn.-Pepin. Wis.

78 White sucker 1.6867

78 White sucker 1.5203

80 Walleye 1.2162

80 White sucker 1.4231

80 White sucker 1.1020

Station 112, Mississippi River at Dubuque, Iowa

78 Common carp 12.9 1.1 1.8868

78 Common carp 14.4 1.5 1.4462

78 Largemouth bass 11.2 1.1 1.5901

80 Black crappic 10.2 0.7 1.6000

80 Common carp 17 2.6 2.2018

80 Common carp 17.8 3 1.4789

Station 113, San Antonio River at McFadden, Tex.

79 Longnose gar 28.5 2.3 0.7767

Station 114, Bear River at Brigham City, Utah

78 Common carp 11.7 0.8 1.5625

78 Common carp 10.2 0.6 0.9957

78 Channel catfish 17.9 2.3 1.3109
Year Species Mean total length, cm Mean total weight, Kg Se, ug/g dry wt.

J-20 Draft November 12, 2004

80 Common carp 17 2.3 1.6667

80 Common carp 12 0.9 1.4583

80 Channel catfish 22.7 4.7 1.2551

Station 115, Colorado River at Yuma, Ariz.- Winterhaven, Calif.

78 Common carp 16 2.7 6.4815

78 Striped mullet 20.3 3.4 3.5958

Station 116, Souris River at Upham, N. Dak.

78 White sucker 16.5 2.3 1.0526

80 Northem pike 19.6 1.9 0.9709

80 White sucker 15.3 1.7 1.1161

80 White sucker 13.8 1.3 1.0331

Station 117, Flathead River at Creston, Mont.

78 Northem squawfish 19.1 2.7 0.7925

80 Largescale sucker 15.4 1.3 0.7589

80 Largescale sucker 15.5 1.3 0.9237

80 Northem squawfish 15.8 1.3 0.9091

Average 1.8836

Std 1.4373

max 10.5204

min 0.3158

count 591
J-21 Draft November 12, 2004

Table J-2. Selenium concentration (μg/g dw whole-body) in fish and invertebrate samples collected at sites

of the USGS National Water Quality Assessment (NAWQA) Program, 1992-1997.

[Se] Scientific Name Common Name Place Name

0.30 Odonata - GOOSE LAKE WMA

0.30 Hydropsyche - SF PALOUSE R. AT ARMSTRONG RD NR PULLMAN, WA

0.30 Hydropsyche - PALOUSE R. AT ENDICOTT-ST. JOHN RD NR COLFAX, WA

0.40 Odonata - JOHNSON WPA

0.40 Odonata - WOOD DUCK WMA

0.40 Corbicula - SALUDA RIVER NEAR COLUMBIA, SC

0.50 Pacifastacus leniusculus signal crayfish TRUCKEE R AT FARAD, CA

0.50 Odonata - DEPARTMENT OF ROADS - ONEILL

0.50 Odonata - TODD VALLEY - MEDUNA SITE

0.50 Micropterus salmoides largemouth bass TRINITY RV BL DALLAS, TX

0.60 Hydropsyche - ROCK CREEK BLW US HWY 30/93 AT TWIN FALLS ID

0.60 Pacifastacus leniusculus signal crayfish TRUCKEE R AT CLARK, NV

0.60 Hydropsyche - WOLF RIVER AT TURTLE LAKE ROAD AT POST LAKE, WI

0.60 Potamogeton pectinatus sago pondweed NORTH BRANCH MILWAUKEE RIVER NR RANDOM LAKE, WI

0.69 Tilapia melanotheron blackchin tilapia ALA WAI CANAL AT HONOLULU, HI

0.70 Hydropsyche - CRAB CREEK AT MORGAN LAKE ROAD NEAR OTHELLO, WA

0.70 Cottus sp. freshwater sculpins MILLER CREEK NEAR DES MOINES, WA

0.70 Cheumatopsyche - WOLF RIVER NEAR POST LAKE, WI

0.70 Hydropsyche - WOLF RIVER NEAR POST LAKE, WI

0.80 Hydropsyche - PESHEKEE RIVER NEAR MARTINS LANDING, MI

0.90 Catostomus clarki desert sucker PINTO CREEK NEAR MIAMI, AZ.

0.90 Odonata - SABATKA SALINE WETLAND

0.90 Corbicula - SYCAMORE CK AT SYCAMORE PK, FT WORTH, TX

0.90 Hydropsychidae net-spinning caddisflies WOLF RIVER AT HIGHWAY M NEAR LANGLADE, WI

1.00 Hydropsyche - SNAKE RIVER AT KING HILL ID

1.00 Cottus sp. freshwater sculpins BIG SOOS CREEK ABOVE HATCHERY NEAR AUBURN, WA

1.00 Orconectes - EAST RIVER AT MIDWAY ROAD NEAR DE PERE, WI

1.00 Hydropsyche - PENSAUKEE RIVER NEAR KRAKOW, WI

1.08 Poecilia sphenops black molly KANEOHE STR BLW KAMEHAMEHA HWY, OAHU, HI

1.10 Acroneuria - WEST BRANCH WHITEFISH RIVER NEAR DIFFIN, MI

1.10 Pacifastacus leniusculus signal crayfish EAST FORK CARSON RIVER NEAR GARDNERVILLE, NV

1.10 Pacifastacus leniusculus signal crayfish EAST FORK CARSON RIVER NEAR DRESSLERVILLE, NV

1.10 Cottus sp. freshwater sculpins SANDY RIVER NEAR TROUTDALE, OR

1.10 Cottus sp. freshwater sculpins GALES CREEK NEAR GLENWOOD, OR

1.10 Cottus sp. freshwater sculpins GALES CREEK NEAR GLENWOOD, OR

1.10 Hydropsyche - PALOUSE RIVER AT HOOPER, WA

1.10 Hydropsyche - SNAKE RIVER AB JACKSON LAKE AT FLAGG RANCH WY

1.10 Hydropsyche - SNAKE RIVER AB JACKSON LAKE AT FLAGG RANCH WY

1.15 Cheumatopsyche - DUCK CREEK AT SEMINARY ROAD NEAR ONEIDA, WI

1.20 Cyprinella lutrensis red shiner GRANITE CREEK AT PRESCOTT, AZ.

1.20 Orconectes causeyi - GRANITE CREEK AT PRESCOTT, AZ.

1.20 Catostomus occidentalis Sacramento sucker COTTONWOOD C NR COTTONWOOD CA

1.20 Xiphophorus helleri green swordtail WAIHEE STR NR KAHALUU, OAHU, HI

1.20 Hydropsyche - HENRYS FORK NR REXBURG ID

1.20 Hydropsyche - ROCK CREEK AB HWY 30/93 XING AT TWIN FALLS ID

1.20 Cottus sp. freshwater sculpins TUALATIN RIVER AT WEST LINN, OR
J-22 Draft November 12, 2004

[Se] Scientific Name Common Name Place Name

1.20 Cottus sp. freshwater sculpins DENNIS C BL BLACK BUTTE MINE, NR COTTAGE GROVE LK

1.20 Cottus sp. freshwater sculpins WEST BRANCH KELSEY CREEK AT BELLEVUE, WA

1.20 Cottus sp. freshwater sculpins BERTRAND CREEK NEAR LYNDEN, WA

1.20 Cheumatopsyche - TOMORROW RIVER NEAR NELSONVILLE, WI

1.20 Hydropsyche - SNAKE RIVER AB JACKSON LAKE AT FLAGG RANCH WY

1.30 Pacifastacus leniusculus signal crayfish TRUCKEE R AT FARAD, CA

1.30 Cottidae sculpins JOHNSON CREEK AT MILWAUKIE, OR

1.30 Micropterus salmoides largemouth bass WHITE ROCK LK IN DALLAS, TX

1.30 Cottus sp. freshwater sculpins DUWAMISH RIVER AT GOLF COURSE AT TUKWILA, WA

1.30 Hydropsyche - DUCK CREEK AT SEMINARY ROAD NEAR ONEIDA, WI

1.39 Cottus cognatus slimy sculpin NINILCHIK R AT NINILCHIK AK

1.40 Hydropsyche - PORTNEUF RIVER AT POCATELLO ID

1.40 Odonata - TRUST - WILD ROSE SLOUGH

1.40 Pacifastacus leniusculus signal crayfish TRUCKEE R AT HWY 447 AT NIXON, NV

1.40 Cottus sp. freshwater sculpins MARYS RIVER AT CORVALLIS, OR

1.40 Cottus sp. freshwater sculpins FIR CREEK NEAR BRIGHTWOOD, OR

1.40 Cottus sp. freshwater sculpins FANNO CREEK AT DURHAM, OR

1.40 Cottus sp. freshwater sculpins FANNO CREEK AT DURHAM, OR

1.40 Cheumatopsyche - DUCK CREEK AT SEMINARY ROAD NEAR ONEIDA, WI

1.40 Hydropsyche - NORTH BRANCH MILWAUKEE RIVER NR RANDOM LAKE, WI

1.50 Catostomus clarki desert sucker SAN PEDRO RIVER AT CHARLESTON, AZ.

1.50 Hydropsyche - ROCK CREEK AB DAYDREAM RANCH NR TWIN FALLS ID

1.50 Brachycentrus - ROCK CREEK AB HWY 30/93 XING AT TWIN FALLS ID

1.50 Odonata - TRUST - MORMON ISLAND CRANE MEADOW, EAST SLOUGH

1.50 Pacifastacus leniusculus signal crayfish CARSON RIVER AT DEER RUN ROAD NEAR CARSON CITY, NV

1.50 Pacifastacus leniusculus signal crayfish CARSON RIVER NEAR FORT CHURCHILL, NV

1.50 Cottidae sculpins MUDDY CREEK NEAR PEORIA, OR

1.50 Hydropsyche - PALOUSE RIVER NEAR COLFAX, WA

1.50 Cottus sp. freshwater sculpins THORNTON CREEK NEAR SEATTLE, WA

1.50 Hydropsyche - TOMORROW RIVER NEAR NELSONVILLE, WI

1.50 Hydropsyche - SALT RIVER AB RESERVOIR NR ETNA WY

1.50 Hydropsyche - SALT RIVER AB RESERVOIR NR ETNA WY

1.53 Cyprinus carpio common carp BEAR RIVER NEAR CORINNE, UT

1.60 Catostomus clarki desert sucker GILA RIVER AT KELVIN, AZ.

1.60 Hydropsyche - BARK RIVER NEAR BARK RIVER, MI

1.60 Cottidae sculpins FANNO CREEK AT DURHAM, OR

1.60 Elliptio - CEDAR CREEK BELOW MYERS CREEK NR HOPKINS, SC

1.60 Hydropsyche - PINE CREEK AT PINE CITY ROAD AT PINE CITY, WA

1.62 Corbicula - TRUCKEE R AT CLARK, NV

1.70 Hydropsyche - PORTNEUF RIVER AT TOPAZ ID

1.70 Hydropsyche - SNAKE RIVER AT KING HILL ID

1.70 Corbicula - TRENT RIVER NEAR TRENTON, NC

1.70 Elliptio - MCTIER CREEK (RD 209) NEAR MONETTA, SC

1.70 Corbicula - GILLS CREEK NEAR HOPKINS,SC

1.70 Hydropsyche - SHEBOYGAN RIVER AT DOTYVILLE, WI

1.79 Cottidae sculpins LITTLE ABIQUA CREEK NEAR SCOTTS MILLS, OR

1.80 Hydropsychidae net-spinning caddisflies TRUCKEE R AT FARAD, CA

1.80 Hydropsyche - TETON RIVER NR ST ANTHONY ID

1.80 Hydropsyche - SNAKE R NR MINIDOKA ID (AT HOWELLS FERRY)

1.90 Corbicula manilensis asian clam CHATTAHOOCHEE R AT SR 253 NEAR CHATTAHOOCHEE, FL

1.90 Hydropsyche - PALOUSE RIVER AT LAIRD PARK NR HARVARD, ID

1.90 Brachycentrus - ROCK CREEK AB HWY 30/93 XING AT TWIN FALLS ID

1.90 Hydropsyche - DUCK CREEK AT SEMINARY ROAD NEAR ONEIDA, WI

1.95 Poecilia sphenops black molly NUUANU STR ABV WAOLANI ST. AT HONOLULU, OAHU, HI

2.00 Hydropsyche - SPRING CREEK AT SHEEPSKIN RD NR FORT HALL ID

2.00 Acroneuria - PESHEKEE RIVER NEAR MARTINS LANDING, MI

2.00 Cheumatopsyche - DUCK CREEK AT SEMINARY ROAD NEAR ONEIDA, WI
J-23 Draft November 12, 2004

[Se] Scientific Name Common Name Place Name

2.03 Ameiurus natalis yellow bullhead SANTA ANA R A HAMNER RD NR NORCO CA

2.05 Richardsonius balteatus redside shiner BEAR RIVER ABOVE RESERVOIR, NEAR WOODRUFF, UT

2.10 Catostomus clarki desert sucker VERDE RIVER ABV W. CLEAR CREEK, NR CAMP VERDE, AZ

2.10 Hydropsyche - SNAKE R NR MINIDOKA ID (AT HOWELLS FERRY)

2.10 Hydropsyche - ROCK CK AT USFS FOOTBRIDGE, NR ROCK CREEK

2.10 Hydropsyche - ROCK CREEK AB HWY 30/93 XING AT TWIN FALLS ID

2.10 Pacifastacus leniusculus signal crayfish TRUCKEE R AT LOCKWOOD, NV

2.10 Hydropsyche - TUALATIN RIVER AT WEST LINN, OR

2.10 Hydropsyche - ESQUATZEL COULEE AT MESA, WA

2.10 Cottus sp. freshwater sculpins FISHTRAP CREEK AT FLYNN ROAD AT LYNDEN, WA

2.17 Corbicula fluminea Asian clam CONTENTNEA CREEK AT HOOKERTON, NC

2.18 Corbicula - TENNESSEE RIVER AT CHATTANOOGA, TN

2.20 Brachycentrus - BITCH CREEK NR LAMONT ID

2.20 Hydropsyche - SOUTH BRANCH PAINT RIVER NEAR ELMWOOD, MI

2.20 Cottus sp. freshwater sculpins LITTLE ABIQUA CREEK NEAR SCOTTS MILLS, OR

2.20 Ceratopsyche - EAST RIVER @ CTH PP IN BROWN COUNTY NR DE PERE, WI

2.20 Brachycentrus - SALT RIVERV NR FISH CK ABOVE SMOOT

2.30 Agosia chrysogaster longfin dace SANTA CRUZ RIVER AT TUBAC, AZ.

2.30 Catostomus clarki desert sucker WEST CLEAR CREEK NEAR CAMP VERDE, AZ.

2.30 Hydropsyche - PORTNEUF RIVER AT TOPAZ ID

2.37 Ameiurus natalis yellow bullhead SANTA ANA R A MWD CROSSING CA

2.40 Xiphophorus helleri green swordtail WAIKELE STR AT WAIPAHU, OAHU, HI

2.40 Hydropsyche - MALAD RIVER NR GOODING ID

2.40 Corbicula - CRABTREE CREEK AT US 1 AT RALEIGH, NC

2.40 Corbicula - BLACKWATER RIVER NEAR FRANKLIN, VA

2.40 Cottus sp. freshwater sculpins ROCK CREEK AT CEDAR FALLS ROAD NEAR LANDSBURG, WA

2.40 Cottus sp. freshwater sculpins JUANITA CREEK AT JUANITA, WA

2.50 Agosia chrysogaster longfin dace SALT RIVER NEAR ROOSEVELT, AZ.

2.50 Hydropsyche - SNAKE RIVER NR BLACKFOOT ID

2.50 Hydropsyche - SNAKE RIVER NR BLACKFOOT ID

2.50 Anaspidacea - EAST FORK CARSON RIVER NEAR GARDNERVILLE, NV

2.50 Elliptio - COOSAWHATCHIE RIVER NR EARLY BRANCH, SC

2.50 Corbicula - TAYLOR FLAT CREEK ABV BIRCH RD NR PASCO, WA

2.52 Xiphophorus helleri green swordtail MANOA STR AT KANEWAI FIELD, HONOLULU, OAHU, HI

2.60 Cottus sp. freshwater sculpins PALMER C AT DAYTON, OR

2.60 Elliptio - SHAWS CREEK NR TRENTON, SC ON CNTY RD 149

2.60 Corbicula - PIGEON RIVER AT NEWPORT, TN

2.60 Corbicula - RUSH CK AT WOODLAND PARK BLVD, ARLINGTON, TX

2.60 Cottus sp. freshwater sculpins NEWAUKUM CREEK NEAR BLACK DIAMOND, WA

2.64 Cyprinus carpio common carp SAN JACINTO R NR ELSINORE CA

2.70 Corbicula - EMORY RIVER AT OAKDALE, TN

2.70 Corbicula - GUADALUPE RV AT GONZALES, TX

2.70 Corbicula - NORTH MEHERRIN RIVER NEAR LUNENBURG, VA

2.70 Cottus sp. freshwater sculpins GREEN RIVER ABOVE TWIN CAMP CREEK NEAR LESTER, WA

2.70 Cottus sp. freshwater sculpins LEACH CREEK NEAR STEILACOOM, WA

2.70 Cottus sp. freshwater sculpins NORTH CREEK BELOW PENNY CREEK NEAR BOTHELL, WA

2.77 Gambusia affinis western mosquitofish MANOA STR AT KANEWAI FIELD, HONOLULU, OAHU, HI

2.79 Cottus sp. freshwater sculpins WEBER RIVER NEAR COALVILLE, UT

2.80 Corbicula manilensis asian clam MUCKALEE CREEK AT GA 195, NEAR LEESBURG, GA

2.80 Corbicula - TAR RIVER NEAR TAR RIVER, NC

2.80 Decapoda crabs PLATTE RIVER AT BRADY, NE (TOTFLO)

2.80 Corbicula - COPPER CREEK NEAR GATE CITY, VA

2.80 Brachycentrus - SECOND SOUTH BRANCH OCONTO RIVER NR MOUNTAIN, WI

2.80 Hydropsyche - TOMORROW RIVER NEAR NELSONVILLE, WI

2.80 Hydropsyche - NORTH BRANCH MILWAUKEE RIVER NR RANDOM LAKE, WI

2.80 Hydropsyche - SNAKE RIVER AB JACKSON LAKE AT FLAGG RANCH WY

2.81 Salvelinus fontinalis brook trout WOOD RIVER ABOVE MIDDLE FORK NEAR MEETEETSE, WY
J-24 Draft November 12, 2004

[Se] Scientific Name Common Name Place Name

2.85 Cottus cognatus slimy sculpin KENAI R AT JIMS LANDING NR COOPER LANDING AK

2.88 Salvelinus fontinalis brook trout CROW CREEK AT MOUTH, AT PAHASKA, WY

2.90 Corbicula manilensis asian clam APALACHICOLA RIVER AT CHATTAHOOCHEE FLA

2.90 Corbicula manilensis asian clam FLINT RIVER AT NEWTON, GA

2.90 Corbicula manilensis asian clam ICHAWAYNOCHAWAY CREEK BELOW NEWTON, GA

2.90 Corbicula manilensis asian clam PEACHTREE CREEK AT ATLANTA, GA

2.90 Corbicula manilensis asian clam FLINT RIVER AT LAKE BLACKSHEAR NEAR WARWICK, GA.

2.90 Corbicula manilensis asian clam CHATTAHOOCHEE RIVER AT COLUMBUS, GA

2.90 Hydropsyche - SNAKE RIVER AT KING HILL ID

2.90 Corbicula - TRUCKEE R AT CLARK, NV

2.96 Cottus cognatus slimy sculpin CHESTER C AT ARCTIC BOULEVARD AT ANCHORAGE AK

2.96 Gambusia affinis western mosquitofish KANEOHE STR BLW KAMEHAMEHA HWY, OAHU, HI

3.00 Cottus cognatus slimy sculpin KENAI R BL RUSSIAN R NR COOPER LANDING AK

3.00 Hydropsyche - PORTNEUF RIVER AT TOPAZ ID

3.00 Brachycentrus - TETON RIVER AB SOUTH LEIGH CREEK NR DRIGGS ID

3.00 Corbicula fluminea Asian clam BIG BLUE RIVER AT SHELBYVILLE, IN

3.00 Corbicula - TAR RIVER AT TARBORO, NC

3.00 Corbicula - NORTH FLAT RIVER AT TIMBERLAKE, NC

3.00 Corbicula - GILLS CREEK AT COLUMBIA, SC

3.10 Perlidae common stoneflies BIG WOOD RIVER BLW BOULDER CK NR KETCHUM

3.10 Corbicula - NEUSE RIVER NEAR COX MILL, NC

3.10 Corbicula - SOUTH FORK CATAWBA RIVER AT MCADENVILLE, NC

3.10 Corbicula - INDIAN CREEK NEAR LABORATORY, NC

3.18 Corbicula manilensis asian clam SNAKE CREEK NEAR WHITESBURG, GA

3.20 Corbicula manilensis asian clam SPRING CREEK NEAR IRON CITY, GA.

3.20 Corbicula - ROANOKE RIVER AT ROANOKE RAPIDS, NC

3.20 Corbicula - NOTTOWAY RIVER NEAR SEBRELL, VA

3.30 Corbicula manilensis asian clam BULL CREEK AT US 27 AT COLUMBUS, GEORGIA

3.30 Corbicula - CONOCOCHEAGUE CREEK AT FAIRVIEW, MD

3.30 Decapoda crabs WOOD RIVER NEAR GRAND ISLAND NEBR

3.30 Corbicula - NOLICHUCKY RIVER NEAR LOWLAND

3.40 Corbicula fluminea Asian clam SUGAR CREEK AT CO RD 400 S AT NEW PALESTINE, IN

3.40 Ceratopsyche - WEST BRANCH WHITEFISH RIVER NEAR DIFFIN, MI

3.40 Cheumatopsyche - JOHNSON CREEK AT MILWAUKIE, OR

3.40 Corbicula - POWELL RIVER NEAR ARTHUR, TN

3.40 Hydropsyche - PARADISE CREEK AT PULLMAN, WA

3.40 Hydropsyche - SALT RIVER AB RESERVOIR NR ETNA WY

3.50 Corbicula manilensis asian clam FLINT R @ 10-MI STILL LANDING NR CHATTAHOOCHEE, FL

3.50 Corbicula manilensis asian clam PEACHTREE CREEK AT ATLANTA, GA

3.50 Corbicula - NEUSE RIVER AT KINSTON, NC

3.50 Corbicula fluminea Asian clam NEUSE RIVER AT KINSTON, NC

3.50 Corbicula - SANTEE R AT TREZESVANTS LANDING NR FT MOTTE, SC

3.50 Corbicula - NOLICHUCKY RIVER AT EMBREEVILLE, TN

3.50 Corbicula - SAN MARCOS RV ABV BLANCO RV BL SAN MARCOS, TX

3.60 Cyprinella lutrensis red shiner SALT RIVER NEAR ROOSEVELT, AZ.

3.60 Agosia chrysogaster longfin dace AGUA FRIA RIVER NEAR ROCK SPRINGS, AZ.

3.60 Carpiodes carpio river carpsucker BUCKEYE CANAL NR HASSAYAMPA

3.60 Corbicula - 48TH STREET DRAIN NR INTERSTATE 10

3.60 Corbicula manilensis asian clam APALACHICOLA RIVER NR BLOUNTSTOWN,FLORIDA

3.60 Corbicula manilensis asian clam CHATTAHOOCHEE R AT SR 369 NR FLOWERY BRANCH, GA.

3.60 Corbicula - SWIFT CREEK AT HILLIARDSTON, NC

3.60 Corbicula - CHICOD CR AT SR1760 NEAR SIMPSON, NC

3.60 Corbicula - LITTLE RIVER NEAR MARYVILLE, TN

3.60 Hydropsyche - CRAB CREEK AT ROCKY FORD ROAD NEAR RITZVILLE, WA

3.60 Cottus sp. freshwater sculpins ROCK CREEK NEAR MAPLE VALLEY, WA

3.60 Cottus sp. freshwater sculpins NOOKSACK RIVER AT BRENNAN, WA
J-25 Draft November 12, 2004

[Se] Scientific Name Common Name Place Name

3.66 Corbicula - CONGAREE RIVER AT COLUMBIA, SC

3.70 Corbicula manilensis asian clam KINCHAFOONEE CREEK NEAR DAWSON, GA

3.70 Hydropsyche - BLACKFOOT RIVER AB RESERVOIR NR HENRY ID

3.70 Corbicula fluminea Asian clam WHITE RIVER AT RAYMOND STREET AT INDIANAPOLIS, IN

3.70 Corbicula manilensis asian clam CURRENT RIVER AT VAN BUREN, MO

3.77 Corbicula - HOLSTON RIVER AT SURGOINSVILLE, TN

3.80 Corbicula manilensis asian clam APALACHICOLA RIVER NR SUMATRA,FLA.

3.80 Corbicula - SWIFT CREEK NEAR APEX, NC

3.80 Cottus sp. freshwater sculpins LUCKIAMUTE RIVER NEAR SUVER, OR

3.80 Corbicula - CATOCTIN CREEK AT TAYLORSTOWN, VA

3.85 Corbicula manilensis asian clam SOPE CREEK NEAR MARIETTA, GA

3.90 Corbicula manilensis asian clam NICKAJACK CR AT COOPER LAKE DR NR MABLETON, GA.

3.90 Corbicula manilensis asian clam CHATTAHOOCHEE RIVER NEAR COLUMBIA, ALA.

4.00 Elliptio - GEORGES CREEK NEAR OLAR, SC ON SC 64

4.00 Corbicula - WATEREE RIVER NR. CAMDEN, SC

4.01 Cyprinus carpio common carp LEON CK AT IH 35 AT SAN ANTONIO, TX

4.10 Corbicula manilensis asian clam WILLEO CREEK AT ST RT 120 NEAR ROSWELL, GA.

4.10 Hydropsyche - MALAD RIVER NR GOODING ID

4.10 Corbicula - CARSON RIVER AT TARZYN ROAD NR FALLON, NV

4.10 Corbicula - FRENCH BROAD RIVER NEAR NEWPORT, TN

4.10 Corbicula - MIDDLE FORK HOLSTON RIVER AT SEVEN MILE FORD, VA

4.16 Corbicula - OBED RIVER NEAR LANCING, TN

4.20 Corbicula manilensis asian clam SEWELL MILL CR AT SEWELL MILL RD NEAR MARIETTA

4.20 Arctopsyche - BIG LOST RIVER AT HOWELL RANCH NR CHILLY ID

4.20 Corbicula - CONGAREE RIVER AT U.S. HWY 601 NR. FORT MOTTE, SC

4.20 Corbicula - GUADALUPE RV NR SPRING BRANCH, TX

4.30 Agosia chrysogaster longfin dace GILA RIVER AT KELVIN, AZ.

4.30 Corbicula manilensis asian clam SOPE CREEK NEAR MARIETTA, GA

4.30 Corbicula manilensis asian clam AYCOCKS CREEK NEAR BOYKIN, GA.

4.30 Elliptio - COW CASTLE CREEK NEAR BOWMAN, SC

4.30 Corbicula - NORTH FORK HOLSTON RIVER NEAR CLOUD FORD, TN

4.40 Cottus cognatus slimy sculpin TALKEETNA R NR TALKEETNA AK

4.40 Corbicula manilensis asian clam COOLEEWAHEE CREEK NEAR NEWTON, GA.

4.40 Corbicula manilensis asian clam SOPE CREEK NEAR MARIETTA, GA

4.40 Corbicula manilensis asian clam FLINT RIVER NEAR CULLODEN, GA

4.40 Corbicula fluminea Asian clam CLIFTY CREEK AT HARTSVILLE, IN

4.40 Pacifastacus leniusculus signal crayfish CARSON RIVER NEAR FORT CHURCHILL, NV

4.47 Corbicula manilensis asian clam CHATTAHOOCHEE RIVER NEAR WHITESBURG, GA

4.50 Agosia chrysogaster longfin dace SAN PEDRO RIVER AT CHARLESTON, AZ.

4.50 Corbicula manilensis asian clam CHATTAHOOCHEE RIVER NEAR NORCROSS, GA

4.50 Arctopsyche - BIG LOST RIVER AT HOWELL RANCH NR CHILLY ID

4.50 Corbicula fluminea Asian clam LOST RIVER NEAR LEIPSIC, IN

4.50 Corbicula - CLINCH RIVER ABOVE TAZEWELL, TN

4.50 Corbicula - ESQUATZEL COULEE AT SAGEMOOR RD NEAR PASCO, WA

4.57 Corbicula - CHAMBERS CK NR RICE, TX

4.60 Corbicula manilensis asian clam FLAT SHOAL CREEK AT STOVALL RD NEAR STOVALL, GA

4.60 Corbicula - BIG LIMESTONE CREEK NEAR LIMESTONE, TN

4.60 Corbicula - SALADO CK AT LOOP 13 AT SAN ANTONIO, TX

4.64 Corbicula - CONGAREE RIVER AT COLUMBIA, SC

4.76 Catostomus commersoni white sucker SADDLE RIVER AT RIDGEWOOD NJ

4.80 Agosia chrysogaster longfin dace PINTO CREEK NEAR MIAMI, AZ.

4.80 Corbicula manilensis asian clam SNAKE CREEK NEAR WHITESBURG, GA

4.80 Corbicula - BEAVER CREEK BELOW LIBERTY HILL, SC

4.80 Cottus sp. freshwater sculpins NF SKOKOMISH R BL STAIRCASE RPDS NR HOODSPORT, WA

4.80 Cottus sp. freshwater sculpins NOOKSACK RIVER AT NORTH CEDARVILLE, WA

4.81 Corbicula - NORTH FORK HOLSTON RIVER NEAR HAYTER GAP, VA

4.86 Cottus cognatus slimy sculpin CHESTER C AT ARCTIC BOULEVARD AT ANCHORAGE AK

5.09 Corbicula - MENARD CK NR FUQUA, TX
J-26 Draft November 12, 2004

5.10 Corbicula manilensis asian clam LIME CREEK NEAR COBB, GA

[Se] Scientific Name Common Name Place Name

5.10 Corbicula fluminea Asian clam KESSINGER DITCH NEAR MONROE CITY, IN

5.10 Corbicula fluminea Asian clam SALT CREEK AT HOOSIER AVENUE AT OOLITIC, IN

5.10 Cottus sp. freshwater sculpins SKOKOMISH RIVER NEAR POTLATCH, WA

5.13 Cottus sp. freshwater sculpins BEAR RIVER BELOW SMITHS FORK, NEAR COKEVILLE, WY

5.19 Cottus cognatus slimy sculpin KAMISHAK R NR KAMISHAK AK

5.20 Cyprinella lutrensis red shiner AGUA FRIA RIVER NEAR ROCK SPRINGS, AZ.

5.20 Corbicula fluminea Asian clam SUGAR CREEK AT CO RD 400 S AT NEW PALESTINE, IN

5.20 Corbicula - SOUTH BRANCH POTOMAC RIVER NEAR SPRINGFIELD, WV

5.30 Corbicula fluminea Asian clam EAST FORK WHITE RIVER AT SHOALS, IN

5.30 Corbicula - EDISTO RIVER NEAR COTTAGEVILLE,SC

5.40 Corbicula manilensis asian clam CHATTAHOOCHEE RIVER NEAR CORNELIA, GA

5.40 Corbicula manilensis asian clam WEST FORK LITTLE RIVER NEAR GAINESVILLE, GA.

5.40 Corbicula - TRUCKEE R AT LOCKWOOD, NV

5.40 Corbicula - BRUSHY CREEK NEAR PELHAM, SC

5.40 Corbicula - BIG CREEK ABOVE SALUDA, SC

5.70 Corbicula - AHOSKIE CR NEAR POORTOWN, NC

5.70 Corbicula - CLINCH RIVER AT SPEERS FERRY, VA

5.78 Hemichromis jewelfishes POAMOHO STREAM NR WAIALUA, OAHU, HI

5.79 Corbicula - KNOB CREEK AT AUSTIN SPRINGS

5.80 Corbicula manilensis asian clam FLINT RIVER NEAR LOVEJOY, GA

5.80 Corbicula - SABINAL RV NR SABINAL, TX

5.81 Cottus cognatus slimy sculpin MOOSE C NR PALMER AK

6.00 Corbicula fluminea Asian clam MUSCATATUCK RIVER NEAR DEPUTY, IN

6.00 Corbicula - LICK CREEK NEAR HOLLAND MILL, TN

6.00 Hydropsyche - CHAFFEE CREEK AT NESHKORO, WI

6.20 Corbicula - MEDINA RV AT LA COSTE, TX

6.30 Corbicula - INDIAN CREEK ABOVE NEWBERRY, SC

6.35 Cottus cognatus slimy sculpin SF CAMPBELL C NR ANCHORAGE AK

6.68 Cottus cognatus slimy sculpin COSTELLO C AB CAMP C NR COLORADO AK

6.70 Agosia chrysogaster longfin dace AGUA FRIA RIVER NEAR MAYER, AZ.

6.70 Agosia chrysogaster longfin dace AGUA FRIA RIVER AT BLOODY BASIN ROAD

6.70 Corbicula manilensis asian clam CHATTAHOOCHEE RIVER NEAR WHITESBURG, GA

6.70 Corbicula - BLANCO RV AT WIMBERLEY, TX

7.00 Corbicula - LONG CREEK ON SPENCER MTN RD NR SPENCER MTN, NC

7.30 Corbicula - GUEST RIVER AT COEBURN, VA

7.70 Corbicula - FRIO RV AT CONCAN, TX

8.10 Corbicula - VERDE R BLW TANGLE CREEK, ABV HORSESHOE DAM, AZ.

8.40 Corbicula - COMAL RV AT NEW BRAUNFELS, TX

8.47 Cottus cognatus slimy sculpin COSTELLO C NR COLORADO AK

9.10 Corbicula - NUECES RV BL UVALDE, TX

9.40 Cyprinella lutrensis red shiner VERDE R BLW TANGLE CREEK, ABV HORSESHOE DAM, AZ.

9.56 Cottus cognatus slimy sculpin CAMP C AT MOUTH NR COLORADO AK

9.83 Ictalurus punctatus channel catfish SABINAL RV NR SABINAL, TX

10.47 Cottus cognatus slimy sculpin COSTELLO C BL CAMP C NR COLORADO AK

12.83 Corbicula - GERONIMO CK AT HWY 90A NR SEGUIN, TX

14.40 Hydropsyche - GREEN CREEK NEAR PALMER, MI

22.37 Salmo trutta brown trout TONGUE RIVER NEAR DAYTON, WY
Ref-1 Draft November 12, 2004

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