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

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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

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## 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 (H2 

### SeO 4 

### ), (+ IV) in selenites (HSeO 3 - 

### , SeO 3 2- 

### ) and selenous acid (H 2 

### SeO 3 

### ), 0 in elemental selenium, and (-II) in selenides (Se 2- 

### , HSe
- 

### ), hydrogen selenide (H 2 

### Se), and organic selenides (R2 

### Se). 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, SeO 3 2- 

### , HSeO 3 - 

### , 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 

### (SeO 4 2-

### /H 2 

### SeO 3 

### ) = 1.15 V; E 0 

### (Cr2 

### O 7 2-

### /Cr 3+ 

### ) = 1.33V; E 0 

### (SO 4 2-

### /H 2 

### SO 3 

### ) = 0.200V (standard potentials in acid solution: Weast 1969)], whereas selenide is a much stronger reducing agent than sulfide [ E 0 

### (Se/H2 

### Se) = -0.36 V; E 0 

### [S/H2 

### S ]= 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 SeO 4 2-

### is comparable to that of SO 4 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 SeO 3 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.

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Most selenite salts are less soluble than the corresponding selenates. The extremely low solubility of ferric selenite Fe2 

### (SeO 3 

### ) 3 

### (Ks 

### = 2.0 ± 1.7 × 10-31 

### ), and of the basic ferric selenite Fe 2 

### (OH) 4 

### SeO 3 

### (K s

### = 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 HSeO 3 -

### or SeO 3 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

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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 75 

### Se 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 SeO 4 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 balthicahas been reported (Luoma et al. 1992). The bioavailability of elemental selenium to M. balthica was assessed by feeding the organisms 75 

### Se-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, H2 

### Se, or other selenides at relatively low redox potentials. Hydrogen selenide by itself is not expected to exist in the aquatic environment since the Se0

### /H 2 

### Se couple falls even below the H +

### /H 2

### couple. Aqueous solutions of H2 

### Se 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 K s

### values of some heavy metal selenides of environmental interest: -11.5 (Mn2+ 

### ), -26.0 (Fe 2+ 

### ), -60.8 (Cu + 

### ), -48.1 (Cu 2+ 

### ), -29.4 (Zn 2+ 

### ), -35.2 (Cd2+ 

### ), and -64.5 (Hg 2+ 

### ). 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

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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 C 

### OH) n

### and humic

substances (dissolved organic matter, DOM). Both sorption to SPM and complexation with colloidal matter reduces the bioavailability of the selenium species. The average fraction of selenium associated with the 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

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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 weatheringprocess 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

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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 (Se O 

### ). 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 Se 0 

### by anaerobic microbial activities. This and water column-derived Se 0

### 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 latipesfrom 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 EC 50 

### , 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 LC 50

### 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 wasconducted 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 LC 50

### values ranged from 215 to 3,020 μg Se/L), but only one flow-through measured acute LC 50 

### 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 LC 50 

### 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 floridaeto 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 LC 50

### 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 LC 50 

### 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 5 th

### 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 LC 50 

### s 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 (F 5,45 

### = 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 LC 50 

### 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).

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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 LC 50

### 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 LC 50

### 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 Ptychocheiluswith 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 LC 50

### 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 LC 50

### 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 LC 50

### 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 LC 50

### 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 CaCO 3 ) 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 1986Table 1a. Acute Toxicity of Selenium to Freshwater Animals (continued)

Species Methoda Chemical Hardness (mg/L as CaCO 3 ) 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 220 d 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 1998Table 1a. Acute Toxicity of Selenium to Freshwater Animals (continued)

Species Methoda Chemical Hardness (mg/L as CaCO 3 ) 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 35,560 f

- 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 >348,320 f

- Hamilton and Buhl 1990b

Chinook salmon (alevin), Oncorhynchus tshawytscha S, U Sodium selenite 41.7 64,690 f

- Hamilton and Buhl 1990bTable 1a. Acute Toxicity of Selenium to Freshwater Animals (continued)

Species Methoda Chemical Hardness (mg/L as CaCO 3 ) 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 34,732 f

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

Species Methoda Chemical Hardness (mg/L as CaCO 3 ) 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 220 d 620

- Kimball, Manuscript Fathead minnow, Pimephales promelas F, M Selenious acid 220 d 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 1995Table 1a. Acute Toxicity of Selenium to Freshwater Animals (continued)

Species Methoda Chemical Hardness (mg/L as CaCO 3 ) 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 1983Table 1a. Acute Toxicity of Selenium to Freshwater Animals (continued)

Species Methoda Chemical Hardness (mg/L as CaCO 3 ) 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 CaCO 3 ) 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. 1985Table 1a. Acute Toxicity of Selenium to Freshwater Animals (continued)

Species Methoda Chemical Hardness (mg/L as CaCO 3 ) 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,bTable 1a. Acute Toxicity of Selenium to Freshwater Animals (continued)

Species Methoda Chemical Hardness (mg/L as CaCO 3 ) 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 1987Table 1a. Acute Toxicity of Selenium to Freshwater Animals (continued)

Species Methoda Chemical Hardness (mg/L as CaCO 3 ) 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 1990bTable 1a. Acute Toxicity of Selenium to Freshwater Animals (continued)

Species Methoda Chemical Hardness (mg/L as CaCO 3 ) 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) 158,422 f 265,990 f

-- 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) >552,000 f >856,083 f

-- Hamilton and Buhl 1990b

Chinook salmon (alevin), Oncorhynchus tshawytscha S, U Sodium selenate 41.7

(sulfate=47) >176,640 f >273,947 f

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

Species Methoda Chemical Hardness (mg/L as CaCO 3 ) 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 329,856 f

-- Buhl and Hamilton 1991

Rainbow trout, Oncorhynchus mykiss F, M Sodium selenate 45

(sulfate=12) 47000 161,168 f

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

Species Methoda Chemical Hardness (mg/L as CaCO 3 ) 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 89,676 f

-- Hamilton 1995

Colorado squawfish (1.7 g juvenile), Ptychocheilus lucius S, U Sodium selenate 196

(sulfate=164) 138358 103,786 f

-- 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 89,572 f

-- 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 17,245 f

-- Hamilton 1995

Bonytail (1.1 g juvenile), Gila elegans S, U Sodium selenate 196

(sulfate=164) 102828 77,134 f

-- Hamilton 1995

Bonytail (2.6 g juvenile), Gila elegans S, U Sodium selenate 196

(sulfate=164) 90706 68,041 f

-- Hamilton 1995

Bonytail (juvenile), Gila elegans S, U Sodium selenate 199

(sulfate=174) 24010 17,401 f

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

Species Methoda Chemical Hardness (mg/L as CaCO 3 ) 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

aS = static; R = renewal; F = flow-through; M = measured; U = unmeasured. bConcentration of selenium, not the chemical. Note: The values underlined in this column were used to calculate the SMAV for the respective species.

cReported by Barrows et al. (1980) in work performed in the same laboratory under the same contract. dFrom Smith et al. (1976).

eCalculated 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. 1981Table 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 1986Table 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 26,300 c

- Chapman 1992

Striped bass (25 d posthatch), Morone saxatilis S, U Sodium selenate 5 23,700 c

- Chapman 1992

Striped bass (31 d posthatch), Morone saxatilis S, U Sodium selenate 5 26,300 c

- Chapman 1992

Striped bass (32 d posthatch), Morone saxatilis S, U Sodium selenate 5 29,000 c

- Chapman 1992

Striped bass (juvenile), Morone saxatilis F, M Sodium selenate 6.0-6.5 85,840 c

- Klauda 1985a,b

Striped bass (prolarvae), Morone saxatilis F, M Sodium selenate 3.5-4.2 9,790 9,790 Klauda 1985a,b

aS = static; R = renewal; F = flow-through; M = measured; U = unmeasured. bConcentration of selenium, not the chemical. Note: The values underlined in this column were used to calculate the SMAV for the respective species.

cNot 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

Rank a 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 6Table 2a. Ranked Freshwater Genus Mean Acute Values (continued)

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

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

Rank a 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

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

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

Rank a 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. bFrom 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 NA c 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 NATable 3a. Ratios of Freshwater Species Mean Acute Values for Selenite and Selenate (continued)

.

Selenite Sensitivity Rank from Table 2a a 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). bFrom Table 1a.
cNA = 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 2b a 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). bFrom 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 organor 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

[Se whole-body

### ] = exp(0.1331 + (0.8937 × ln[Se muscle 

### ])) (I)
[Se whole-body

### ] = 0.0173 + (0.4634 × [Se ovary 

### ]) (II)
[Se whole-body

### ] = -0.2609 + (0.3071 × [Se liver

### ]) (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.

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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 (EC 20 

### ) 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).

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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 EC 20

### 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 xsymbolizes the selenium concentration in the organism’s tissues, yis the response of interest (survival, growth, or reproduction), andy 0 

### , aand bare model parameters estimated by the regression analysis. The

y 0 

### 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 EC 20

### 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

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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 themost 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

.

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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/gdw) 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 EC 20

### 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

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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 EC 20

### value, however, was computed for the relationship between craniofacial deformities and the concentration of

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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 124weeks 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

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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-

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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

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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 EC 20

### 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

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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 EC 20

### 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

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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 10 5 

### /length

3 

### ) 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 4 O 

### C and 20O 

### C with biological and selenium measurements made every 60 days. Survival and whole-body lipid content were unaffected at 20 O 

### C(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 4 O 

### C exhibited significantly elevated mortality (40.4 percent) relative to controls(2.9 percent), and exhibited significantly greater oxygen consumption and reduced lipid content, which are all indicative of an additional stress load. The chronic value for juvenile bluegill sunfish exposed to waterborne and dietary selenium at 4 O 

### C 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 20 O 

### C 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/gdw (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 EC 20

### of 44.57 μg/g dw specific for fish from Belews Lake, NC, again substantially above the GMCV of 9.500 μg/gdw. 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 dwto 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 O 2

### 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 to8.84 m g 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 dw a 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 EC 20for 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 EC 20for 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 EC 20for 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 11.65 b (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 19.16 b (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: EC 20for craniofacial deformities 5.79 c (parent tissue) 5.85 c (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 >10.92 c (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 dw a 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 13.2 c (parent tissue)
12.4 c (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 51.40 d (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 5.961 d (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 <73 d (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 <18.21 e (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 texanusrazorback 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 >23.28Species Reference Exposure route Selenium form Toxicological endpoint Chronic value, μg/g dw a SMCV

μg/g dw GMCV

μg/g dw



## 80 Draft November 12, 2004 Xyrauchen texanusrazorback 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 <59.92 c,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 <43.70 c,d >20.29 c,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 >5.45 c,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 <28.20 d (larval tissue); or <21.47 d,e (parent tissue) Lepom is macrochirus bluegill Coyle et al. 1993 dietary and waterborne (lab) diet: seleno-Lmethionine water: 6:1 selenate:selenite EC 20for 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 4 o C <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 20

o C >6.0 d

(juvenile tissue)Species Reference Exposure route Selenium form Toxicological endpoint Chronic value, μg/g dw a 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 >3.74 d (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 >17.35 d (parent tissue)

Centrarchidae (9 species) Lemly 1993b dietary and waterborne (field - Belews Lake, NC) not determined EC 20for 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 <14.75 c (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, aState 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 LC 50

### 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 EC 50

### for the saltwater diatom, Skeletonema costatum , is 7,930 μg/L, based on reduction in chlorophyll a (Table B-1). Growth of Chlorellasp., 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 ofChrysochromulina 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 Chorellasp., 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 CaCO 3 ) 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 15,000 b Kumar and Prakash 1971
Blue-green alga, Anacystis nidulans Sodium

selenite - 10-18 LC50 30,000 b Kumar and Prakash 1971
Blue-green alga, Microcystis aeruginisa Sodium

selenite - 8 Incipient inhibition 9,400

(9,300) Bringmann and

Kuhn 1976; 1978a,bTable B-1. Toxicity of Selenium to Aquatic Plants (cont.)

Species Chemical Hardness (mg/L as CaCO 3 ) 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 andJanssenMommen 1993 Duckweed, Lemna minor Sodium

selenite - 14 NOEC (mult. rate) 800 Jenner andJanssenMommen 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 39,000 b Kumar and Prakash 1971Table B-1. Toxicity of Selenium to Aquatic Plants (cont.)

Species Chemical Hardness (mg/L as CaCO 3 ) Duration

(days) Effect Concentration (μg/L) a Reference



## B-6 Draft November 12, 2004 Blue-green alga, Anabaena viriabilis Sodium

selenate - 10-18 EC50 17,000 b 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 andJanssenMommen 1993 Duckweed, Lemna minor Sodium

selenate - 14 NOEC (mult. Rate) >2,400 Jenner andJanssenMommen 1993Table 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 acid c - 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 1991aTable 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. bEstimated from published graph.
cReported 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 theeffect 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 magnaexposed 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 Daphniato bluegills) concentration factors (CFs) for the transfer of selenium from algae to Daphniaand 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 orabnormally 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 ) BCF b

(L/kg) BAF b (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 10 c Adams 1976

Rainbow trout, Oncorhynchus mykiss Sodium

selenite 100 28 Whole-body (2.3 μg/g ) 23 Gissel-Nielsenand 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 52 d Bertram and Brooks 1986

Fathead minnow (6-9 mo.), Pimephales promelas Sodium

selenate 21.5 56 Whole-body 26 d Bertram and Brooks 1986

Fathead minnow (6-9 mo.), Pimephales promelas Sodium

selenate 43.5 56 Whole-body 21 d 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 ) BCF b

(L/kg) BAF b (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 550 e 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 Natura l 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/A g(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 ) BCF b

(L/kg) BAF b (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 ) BCF b

(L/kg) BAF b

(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

4 570 e 293 e 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. 1999Table C-2 Continued.

Other Species Selenium Form Concentration

in Water (μg/L) a Duration

(days) Tissue

(Concentration ) BCF b

(L/kg) BAF b

(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 SO 4

### 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 LC 50

### values of two desmids ( Cosmeriumspp.) exposed to selenate plus sulfate were 4x and 8x the LC 50

### values of selenate only (Sarma and Jayaraman 1984). The growth of Selenastrum capricornutumincreased by 50 percent when 11 or 107 μg/L Se(VI) were combined with 3.3 or 33 μg/L sulfate (Williams et al. 1994). The toxicity of 490 μg/L selenate to D. magnawas 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 SO 4

### 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 reinhardtiiresponded less to sulfate and Se(VI) coexposure (Williams et al. 1994) than Selenastrum capricornutum(7 percent low SO 4

### 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 Daphniashow no interaction of Se and SO4

### in a microcosm experiment (Besser et al. 1989). However, other experiments with Se and SO 4

### 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 SO 4

### (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. LC 50

### value differences due to hardness were no less than half or more than double the LC 50

### of the standard of comparison.
D. magnawere exposed to three forms of selenium and one Se mixture in acute toxicity tests (48h LC 50

### ) to determine the effect of soft (46 mg/L CaCO 3 

### ) 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 (LC 50 

### , hard/soft = 0.5), as was the 1:1 mixture of Se(IV) and Se(VI) (LC 50 

### , hard/soft = 0.6) (Ingersoll et al. 1990). Mytilus eduliswere 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 CaCO 3 

### ) 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 CaCO 3 

### ) water (LC 50

### hard/soft = 1.8) with Se(IV) in soft water being more toxic. The LC50

### of Se(IV) in 1 ‰ saline (455 mg/L CaCO 3 

### ) 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 CaCO 3 

### ) 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 dubiaat 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 HgCl 2

### (5 μg/L) (Pelletier 1986a). Uptake of Se in a 96-hr study (Se(IV), 2 μg/L; HgCl

2 

### , 0.1- 1.0 mg/L) decreased as a function of Hg concentration (r 2

### = - 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 ( Crypthecodiniumsp., 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 byMytilus 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, Q O2

### 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 anumber 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.

### Draft November 12, 2004 F-1 APPENDIX F OTHER DATA

### Draft November 12, 2004 F-2 Other Data

SeleniteAdditional 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. magnato selenate for seven days and obtained an LC 50

### of 1,870 ?



### g/L. This value is in the range of the 48-hr EC 50 

### s 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 LC 50

### 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.

### Draft November 12, 2004 F-3 Table F-1. Other Data on Effects of Selenium on Aquatic Organisms

Species Chemical Hardness (mg/L as CaCO 3 ) Duration Effect Concentration a 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 = 12-21 b 10-100 Foe and Knight, Manuscript
Green alga, Selenastrum capricornutum Sodium selenite - 72 hr BCF = 11,164 c 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 CaCO 3 ) Duration Effect Concentration a Reference



### Draft November 12, 2004 F-4 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 220 d 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, Cyclocyprissp. 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 CaCO 3 ) Duration Effect Concentration a Reference



### Draft November 12, 2004 F-5 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-Nielsenand 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-Nielsenand 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 CaCO 3 ) Duration Effect Concentration a Reference



### Draft November 12, 2004 F-6 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 55.2 e 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 CaCO 3 ) Duration Effect Concentration a Reference



### Draft November 12, 2004 F-7 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 220 d 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 CaCO 3 ) Duration Effect Concentration a Reference



### Draft November 12, 2004 F-8 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 CaCO 3 ) Duration Effect Concentration a Reference



### Draft November 12, 2004 F-9 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 CaCO 3 ) Duration Effect Concentration a Reference



### Draft November 12, 2004 F-10 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 - growthChronic valuegrowthChronic 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-Lmethionine 16 323 days MATC larval survival (dietary only exposure) 20.83

μg Se/g dw (food) Woock et al.

1987

Bluegill (juvenile), Lepomis macrochirus Seleno-Lmethionine 283 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 CaCO 3 ) Duration Effect Concentration a Reference



### Draft November 12, 2004 F-11 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, Chlorellasp. 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



### Draft November 12, 2004 F-12 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, Chlorellasp. Sodium

selenate 32 14 days No effect on rate of cell 10-1,000 Wheeler et al. 1982 Green alga, Chlorellasp. 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



### Draft November 12, 2004 F-13 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. bConverted from dry weight to wet weight basis (see Guidelines)
cGrowth of algae was inhibited dFrom Smith et al. (1976).
eCalculated from the published data using probit analysis and allowing for 8.9% spontaneous mortality.

### Draft November 12, 2004 F-14 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,

### Draft November 12, 2004 F-15 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

### Draft November 12, 2004 F-16 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.

### Draft November 12, 2004 F-17 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

### Draft November 12, 2004 F-18 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 shrimpand 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 magnabased 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+ bln([Se] Tissue

### )) where aand bare 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 gis 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)) + 2

k

### 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 (nis the sample size). In such instances, a second-order version of AIC, AIC c 

### , is recommended (Hurvich and Tsai 1989):

AIC c

### = -2 ln(‹ 

### (parameters|data)) + 2 k
Hurvich and Tsai (1990) demonstrated that the modified version of AIC c

### for least absolute deviation(L1AIC c 

### ) 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 AIC c

### 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 ) i 

### is the difference in AIC (AIC c 

### ) between model iand the model with the lowest AIC (AIC c 

### ) value. Weights for all candidate models sum to 1. For each model, we computed the sum of weighted absolute deviations (SWAD), AIC c

### 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 (R 1 

### ), 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 (AIC c 

### ), differences between the model AIC c

### and the lowest AIC c

### of all candidate models (Delta), weight (w), rank (by weight), and coefficient of determination (R 1 

### ) 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 R 1 

### [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 R 1 

### [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 R 1 

### [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+ b ln([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

## I-1 Draft November 12, 2004 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 (Na 2 

### SeO 4 

### ) 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 × 10 6

### 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. calyciflorushave 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 (EC 20 

### ) 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 EC 20

### values Survival (after 60 d of exposure) Growth (after 60 d of exposure) Tissue Se (μg/g dw) Tissue Se (μg/g dw) SLD NA a 

### 15.74 SeMet NA a 

### 10.47

a 

### The EC 20

### 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. EC 20

### 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 13.08μg/g dw. EC 20

### 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 19.16μg/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 26 th

### 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 EC 20

### 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: EC 20

### 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, EC 20

### 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 EC 20

### 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 ± 3.7 b 

### 0.2 ± 0.2c 

### 14.7 ± 3.4 11.7 ± 2.7 10.6 ± 1.9 12.0 ± 4.1 % SK 13.8 ± 5.6b 

### 0.7 ± 0.5 c 

### 19.4 ± 8.2 11.1 ± 2.3 15.6 ± 4.7 4.9 ± 4.9 % FF 3.2 ± 2.0b 

### 0.2 ± 0.2c 

### 6.8 ± 3.0 15.5 ± 6.6 4.0 ± 0.9 1.5 ± 0.2 % ED 30.8 ± 2 7.4 b 

### 0.2 ± 0.2c 

### 19.9 ± 8.5 13.9 ± 5.3 10.8 ± 2.5 7.5 ± 0.4 % TD 38.9 ± 2 5.6 b 

### 0.7 ± 0.5 c 

### 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 ± 3.5 b 

### 3.0 ± 0.5 c 

### 5.6 ± 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 ± 2.4c 

### 0.1 ± 0.1 b 

### % ED 0.6 ± 0.4 0.3 ± 0.1 0.4 ± 0.1 0.3 ± 0.2 1.7 ± 1.2 % TD 14.4 ± 3.6b 

### 4.0 ± 2.3 c 

### 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. 

## EC 20

## 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: [Se muscle 

### ] = 0.1827 + 0.1287[Se egg 

### ] (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 ([Se muscle 

### ] = 0.2613 + 0.1418[Se egg 

### ]) 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 ; EC 20

### analysis not appropriate for data sets Brook trout 2000: no value available ; EC 20

### analysis not appropriate for data sets Rainbow trout 2001: EC 20

### 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 ; EC 20

### 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 (H 2 75 

### SeO 4 

### ) 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 75 

### Se 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 75 

### Se 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

aSignificantly different from the control. bValues 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 F 1 

### 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, F 2

### 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 EC 20

### 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 (1 st

### factor - selenium; 2 nd

### 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 (1 st

### factor - test water type; 2 nd

### 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 werereplicated 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 weremeasured 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 25 o 

### C) was based on the number of yolk-sac larvae. In 1982, about 200 embryos from 8 crosses were observed and preserved at intervals up to 40 h 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 EC 20

### 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 10 5 

### /length 3 

### ), were measured at selected intervals. Effects Data : The waterborne exposure (see table below) was determined to have an EC 20

### = 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 O 2 

### 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 Se a 

### Surv. % lipid, % O 2 b 

### Se a 

### Surv. % lipid, % O 2 b 

### Se a 

### Surv. % lipid, % O 2 b 

### Se a 

### Surv. % lipid, % O 2 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 I i

### = [ 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, 10 6/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, 10 4/ml 0.95 0.86 0.97 2.05* 0.83 2.38* 0.91 2.30* total leucocytes, 10 4/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, 10 6/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, 10 4/ml 0.86 0.84 0.83 2.30* 0.89 2.49* 0.90 2.36 total leucocytes, 10 4/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 BG a

### 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 II a,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 III a 

### 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 CaCO 3 

### ) 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 EC 20

### 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 EC 20

### 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 EC 20

### 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 seleniu m 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 bod y [μg/g dw]Cumulative proportion 

### National Contaminant Biomonit. Pgm (fish

samples) n = 591 USGS NAWQA Sites (fish and in vertebrate 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 0.7104Year 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 1.1885Year 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 3.3754Year 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 1.4126Year 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 1.0121Year 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 1.9454Year 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 1.5830Year 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 1.5113Year 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 0.9884Year 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 6.9466Year 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 0.7692Year 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 2.5556Year 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 1.3109Year 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

### Draft November 12, 2004 Ref-1 REFERENCES Abdel-Moati, A.R. and M.M. Atta. 1991. Patella vulgata , Mytilus minimusand Hyal prevostias bioindicators for lead and selenium enrichment in Alexandria coastal waters. Mar. Pollut. Bull. 22(3):148-150. Adams, W.J. 1976. The toxicity and residue dynamics of selenium in fish and aquatic invertebrates. Ph.D. thesis. Michigan State University, East Lansing, MI. Available from University Microfilms, Ann Arbor, MI. Order No. 76-27056. Adams, W.J. and B.B. Heidolph. 1985. Short-cut chronic toxicity estimates using Daphnia magna . In: Aquatic toxicology and hazard assessment: Seventh symposium. Cardwell, R.D., R. Purdy and R.C. Bahner (Eds.). ASTM STP 854. American Society for Testing and Materials. Philadelphia, PA. pp. 87- 103. Adams, W.J. and H.E. Johnson. 1981. Selenium: A hazard assessment and a water quality criterion calculation. In: Aquatic toxicology and hazard assessment: Fourth syposium. Branson, D.R. and K.L. Dickson (Eds.). ASTM STP 737. American Society for Testing and Materials, Philadelphia, PA. pp. 124- 137. Adams, W.J., K.V. Brix, K.A. Cothern, L.M. Tear, R.D. Cardwell, A. Fairbrother and J.F. Toll. 1998. Assessment of selenium food chain transfer and critical exposure factors for avian wildlife species: Need for site-specific data. In: Environmental Toxicology and Risk Assessment: Seventh vol., E.E. Little, A.J. DeLanny and B.M. Greenberg (Eds.). ASTM STP 1333. American Society for Testing and Materials, Philadelphia, PA. Adeloju, S.B. and T.M. Young. 1994. Cathodic stripping potentiometric determination of selenium in biological and environmental materials. Anal. Chim. Acta 296(1):69-76. Aguirre, A.A., G.H. Balazs, B. Zimmer man and F.D. Galey. 1994. Organic contaminants and trace metals in the tissues of green turtles ( Chelonia mydas ) afflicted with fibropapillomas in the Hawaiian islands. Mar. Pollut. Bull. 28(2):109-114. Ahsanullah, M. and G.W. Brand. 1985. Effect of selenite and seleniferous fly-ash leachate on growth and viability of the marine amphipod Allorchestes compressa . Mar. Biol. 89:245-248. Ahsanullah, M. and D.H. Palmer. 1980. Acute toxicity of selenium to three species of marine invertebrates, with notes on a continuous-flow test system. Aust. J. Mar. Freshwater Res. 31:795-802. 

## Akaike, H. 1973. Information theory as an extension of the maximum likelihood principle. In B. N. Petrov and F. Csaki, (Eds). Second international symposium on information theory. Akademiai Kiado, Budapest. pp. 267-281

. Akesson, B. and T.S. Srikumar. 1994. Occurrence of low-molecular-weight and high-molecular-weight selenium compounds in fish. Food Chem. 51(1):45-49.

### Draft November 12, 2004 Ref-2 Aksnes, A., K.E. Gulbrandsen and K. Julshamn. 1983. Contents and biological availability of selenium in oxidized and protected fish meals from mackerel. Fiskeridir. Skr., Ser. Ernaer. 2(4):117-24. Al-Sabti, K. 1994. Micronuclei induced by selenium, mercury, methylmercury and their mixtures in binucleated blocked fish erythrocyte cells. Mutat. Res. 320(1-2):157-163. Al-Sabti, K. 1995. An in vitro binucleated blocked hepatic cell technique for genotoxicity testing in fish. Mutat. Res. 335(2):109-120. Alaimo, J., R.S. Ogle and A.W. Knight. 1994. Selenium uptake by larval Chironomus decorusfrom a Ruppia maritima -based benthic/detrital substrate. Arch. Environ. Contam. & Toxicol. 27(4):441-448. Albers, P.H., A.A. Belisle, D.M. Swineford, and R.J. Hall. 1985. Environmental contamination in the oil fields of western Pennsylvania. Oil Petrochem. Pollut. 2(4):265-80. Albers, P.H., D.E. Green, and C.J. Sanderson. 1996. Diagnostic criteria for selenium toxicosis in aquatic birds: Dietary exposure, tissue concentrations, and macroscopic effects. J. Wild. Dis. 32(3):468-485. Albertano, P. And G. Pinto. 1986. The action of heavy metals on the growth of three acidophilic algae. Bull. Soc. Nat. Napoli, 95:319-28. Allaway, W.H., E.E. Cary and C.F. Ehlig. 1967. The cycling of low levels of selenium in soils, plants, and animals. In: Selenium in biomedicine. Muth, O.H. (Ed.). Avi Publishing Company, Westport, CT. pp. 273- 296. Allen, G.T. and R.M. Wilson. 1990. Selenium in the Aquatic Environment of Quivira National Wildlife Refuge Kansas USA. Prairie Naturalist 22(2):129-135. Allen, K.N. 1991. Seasonal Variation of Selenium in Outdoor Experimental Stream-wetland Systems. J. Environ. Qual. 20(4):865-868. Amaratunga, W. and J.B Milne. 1994. Studies on the interaction of selenite and selenium with sulphur donors: Part 2. A Kinetic Study of the Reaction with 2-Mercaptoethanol. Canadian Journal of Chemistry. 72: 2506- . Ambulkar, M. N., V.V.S. Ramakrishna and A.N. Garg. 1995. An environmental pollution study of toxic elements in marine fish from east coast by neutron activation analysis. NUCAR 95: Proc. Nucl. Radiochem. Symp., 417-418. Amiard J.C., D. Pain and H.T. Delves. 1991. Exposure to Trace Elements of Flamingos Living in a Biosphere Reserve the Camargue France. Environ. Pollut. 69(2-3):193-202. Amiard, J.C., B. Berthet and S. Boutaghou. 1993. Seasonal selenium variations in mussels and oysters from a French marine farm. J. Food Comp. Anal. 6(4):370-380. Andersen, J. L. and M.H. Depledge. 1997. A survey of total mercury and methylmercury in edible fish and invertebrates from Azorean waters. Mar. Environ. Res., 44(3):331-350.

### Draft November 12, 2004 Ref-3 Andreev, G. And V. Simeonov. 1992. Interphase distribution and accumulation of elements in the marine environment of the Black Sea. Toxicol. Environ. Chem. 36(1-2):99-104. Angulo, E. 1996. The Tomlinson Pollution Load Index applied to heavy metal, 'Mussel-Watch' data: a useful index to assess coastal pollution. Sci. Total Environ. 187(1):19-56. Apte, S.C., A.G. Howard, R.J. Morris and M.J. McCartney. 1987. Arsenic, antimony and selenium speciation during a spring phytoplankton bloom in a closed experimental ecosystem. Mar. Chem. 20(2):119-130. Arruda, M.A.Z., M. Gallego and M. Valcarcel. 1996. Semi-on-line microwave-assisted digestion of shellfish tissue for the determination of selenium by electrothermal atomic absorption spectrometry. J. Anal. Atomic Spectrom. 11(2):169-173. Arvy, M.P., M. Thiersault and P. Doireau. 1995. Relationships between selenium, micronutrients, carbohydrates, and alkaloid accumulation inCatharanthus roseuscells. J. Plant Nutr. 18(8):1535-1546. Arway, J.A. 1988. The environmental significance of toxic metals found in fish collected throughout the coalfields of Pennsylvania. Inf. Circ. - U. S., Bur. Mines, IC 9184, Mine Drain. Surf. Mine Reclam., Vol. 2, 154-60. Ashton, A. 1991. 'Oyster-water' for monitoring coal ash lagoons in an environmentally sensitive area of Hong Kong. Mar. Pollut. Bull. 22(7):334-339. Audas, A., G.R. Hogan and H. Razniak. 1995. Incubation temperature as a modifying factor on survival of Tenebrio molitor reared in selenium-containing media. J. of Toxicol. & Environ. Health 44(1):115-122. Augier, H., C. Ronneau, P. Roucoux, R. Lion and O. Charlent. 1991. Neutron-activation Analysis of the Elementary Composition of the Marine Phanerogam posidonia-oceania from a Reference Area in Port Cros National Park French Mediterranean. Mar. Biol. (Berlin) 109(2):345-353. Augier, H., L. Benkoel, J. Brisse, A. Chamlian and W.K. Park. 1993a. Microscopic localization of mercury - selenium interaction products in liver, kidney, lung and brain of mediterranean striped dolphins (Stenella coeruleoalba) by silver enhancement kit. Cell. Mol. Biol. (Paris), 39(7):765-772. Augier, H., L. Benkoel, A. Chamlian, W.K. Park and C. Ronneau. 1993b. Mercury, Zinc And Selenium Bioaccumulation In Tissues And Organs Of Mediterranean Striped Dolphins Stenella coeruleoalba Meyen, Toxicological Result Of Their Interaction. Cell. Mol. Biol. (Noisy-Le-Grand) 39(6):621-634. Augier, H., W.K. Park and C. Ronneau. 1995a. Neutron activation study of the elementary composition of edible sea urchins (

Paracentrotus lividusLamarck) in marine creek area polluted by city sewages of Marseille (France). Cell. Mol. Biol. (Paris) 41(4):533-544. Augier, H., J.M Harmand, and C. Ronneau. 1995b. Neutron activation study of the natural elementary composition of edible sea urchins ( Paracentrotus lividusLamarck) in the National Park of Port-Cros (Mediterranean, France). Cell. Mol. Biol. (Noisy-Le-Grand) 41(4):525-531.

### Draft November 12, 2004 Ref-4 Augspurger, T., J.C. Franson, K.A. Converse, P.R. Spitzer, and E.A. Miller. 1998. An epizoatic of common loons in coastal waters of North Carolina: Concentrations of elemental contaminants and results of necropsies. Environ. Toxicol. Chem. 17(2):205-209. Avery, E.L., R.H. Dunstan and J.A. Nell. 1996. The detection of pollutant impact in marine environments: condition index, oxidative DNA damage, and their associations with metal bioaccumulation in the Sydney rock oyster Saccostrea commercialis . Arch. Environ. Contam. Toxicol. 31(2):192-198. Azaizeh, H.A., S. Gowthaman, and N. Terry. 1997. Microbial selenium volatilization in rhizosphere and bulk soils from a constructed wetland. J. Environ. Quality 26(3):666-672. Baatrup, E. 1989. Selenium-induced autometallographic demonstration of endogenous zinc in organs of the rainbow trout, Salmo gairdneri . Histochem. 90(6):417-426. Baatrup, E. and G. Danscher. 1987. Cytochemical demonstration of mercury deposits in trout liver and kidney following methylmercury intoxication: differentiation of two mercury pools by selenium. Ecotoxicol. Environ. Safety 14(2):129-41. Baatrup, E., M. G. Nielsen and G. Danscher. 1986. Histochemical demonstration of two mercury pools in trout tissues: Mercury in kidney and liver after mercuric chloride exposure. Ecotoxicol. Environ. Safety 12(3):267-282. Babich, H., J.A. Puerner and E. Borenfreund. 1986. In vitro cytotoxicity of metals to bluegill (BF-2) cells. Arch. Environ. Contam. Toxicol. 15(1):31-7. Babich, H., N. Martin-Alguacil, and E. Borenfreund. 1989. Arsenic-selenium interactions determined with cultured fish cells. Toxicol. Letters (Amsterdam) 45(2-3):157-164. Bacon, M. and W.J. Ingledew. 1989. The Reductive Reactions of Thiobacillus Ferrooxidans on Sulfur and Selenium. FEMS Microbiology Letters. 58: 189- . Badsha, K. S. and C.R. Goldspink. 1988. Heavy metal levels in three species of fish in Tjeukemeer, a Dutch polder lake. Chemosphere 17(2):459-63. Baer, K.N., D.G. Hutton, R.L. Boeri, T.J. Ward and R.G. Stahl Jr. 1995. Toxicity evaluation of trap and skeet shooting targets to aquatic test species. Ecotoxicol. 4(6):385-392. Bailey, F.C., A.W. Knight, R.S. Ogle and S.J. Klaine. 1995. Effect of sulfate level on selenium uptake by Ruppia maritima . Chemosphere 30(3):579-591. Baines, S.B. and N.S. Fisher. 2001. Interspecific Differences in the Bioconcentration of Selenite by Phytoplankton and Their Ecological Implications. Marine Ecology: Progress Series. 213, 1- . Baker, R.T.M. and S.J. Davies. 1997. The quantitative requirement for alpha.-tocopherol by juvenile African catfish, Clarias gariepinus Burchell. Anim. Sci. 65(1):135-142. Baker, W.B., Jr., S.M. Ray and A.M. Landry, Jr. 1991. Investigation of coal combustion byproduct

### Draft November 12, 2004 Ref-5 utilization for oyster reef development in Texas Bay waters. Proc. - Int. Ash Use Symp., 9th, Volume GS-7162, Vol. 2, 48/1-48/14. Electric Power Research Institute, Palo Alto, CA Baldwin, S. and W. Maher. 1997. Spatial and temporal variation of selenium concentration in five species of intertidal molluscs from Jervis Bay, Australia. Mar. Environ. Res. 44(3):243-262. Baldwin, S., W. Maher, E. Kleber, and F. Krikowa. 1996. Selenium in marine organisms of seagrass habitats (Posidonia australis ) of Jervis Bay, Australia. Mar. Pollut. Bull. 32(3):310-316. Barghigiani, G., D. Pellegrini, A. Dulivo and S. DeRanieri. 1991. Mercury Assessment and its Relation to Selenium Levels in Edible Species of the Northern Tyrrhenian Sea. Mar. Pollut. Bull. 22(8):406-409. Barghigiani, G., A. Dulivo, R. Zamboni and L. Lampugnani. 1993. Interaction Between Selenium and Cadmium in Eledone cirrhosaof the Northern Tyrrhenian Sea. Mar. Pollut. Bull. 26(4):212-216. Bariaud, A. and J.C. Mestre. 1984. Heavy metal tolerance in a cadium-resistant population of Euglena gracilis . Bull. Environ. Contam. Toxicol. 32:597-601. Baron, L.A., T.L. Ashwood, B.E. Sample and C. Welsh. 1997. Monitoring bioaccumulation of contaminants in the belted kingfisher ( Ceryle alcyon). Environ. Monitor. Assess. 47:153-165. Barrington, J. W., A. Jones, D. James, S. Smith and T.P. Stephenson. 1997. Antioxidant deficiency following clam enterocystoplasty. Br. J. Urol. 80(2): 238-242. Barrows, M.E., S.R. Petrocelli, K.J. Macek and J.J. Carroll. 1980. Bioconcentration and elimination of selected water pollutants by bluegill sunfish ( Lepomis macrochirus). In: Dynamics, exposure and hazard assessment of toxic chemicals. Hague, R. (Ed.). Ann Arbor Science Publishers, Ann Arbor, MI. pp. 379- 392. Batley, G.E. 1987. Heavy metal speciation in waters, sediments and biota from Lake Macquarie, New South Wales. Aust. J. Mar. Freshwater Res. 38(5):591-606. Baumann, P.C. and R.B. Gillespie. 1986. Selenium bioaccumulation in gonads of largemouth bass (Micropterus salmoides) and bluegill ( Lepomis macrochirus) from three power plant cooling reservoirs. Environ. Toxicol. Chem. 5(7):695-702. Baumann, P.C. and T.W. May. 1984. Selenium residues in fish from inland waters of the United States. In: Workshop proceedings: The effects of trace elements on aquatic ecosystems. Electric Power Research Institute, Palo Alto, CA. pp. 7-1 to 7-16. Beauchamp, J. J., and J. S. Olson. 1973. Corrections for bias in regression estimates after logarithmic transformation. Ecology. 54:1403-1407. Beal, A.R. 1974. A study of selenium levels in freshwater fishes of Canada's central region. Technical Report Series No. CEN/T-74-6. Environment Canada. Beck, K.M., P. Fair, W. Mcfee, and D. Wolf. 1997. Heavy metals in livers of bottlenose dolphins stranded

### Draft November 12, 2004 Ref-6 along the South Carolina Coast. Mar. Pollut. Bull. 34(9):734-739. Becker, K.B., M.J. Schneider, J.C. Davey, V.A. Galton. 1995a. The type III 5-deiodinase in Rana catesbeiana tadpoles is encoded by a thyroid hormone-responsive gene. Endocrinol. 136(10):4424-4431. Becker, P.R., E.A. Mackey, R. Demiralp, R. Suydam, G. Early, B.J. Koster and S.A. Wise. 1995b. Relationship of silver with selenium and mercury in the liver of two species of toothed whales (Odontocetes). Mar. Pollut. Bull. 30(4):262-271. Beijer, K. and A. Jernelov. 1978. Ecological aspects of mercury-selenium interactions in the marine environment. Environ. Health Perspect. 25:43-45. Beland, P., S. DeGuise, C. Girard, A. Lagace, D. Martineau, R. Michaud, D.C.G. Muir, R.J. Norstrom, E. Pelletier, et al. 1993. Toxic compounds and health and reproductive effects in St. Lawrence beluga whales. J. Great Lakes Res. 19(4):766-775. Beliaeff, B., T.P. O'Connor, D.K. Daskalakis and P.J. Smith. 1997. U.S. mussel watch data from 1986 to 1994: temporal trend detection at large spatial scales. Environ. Sci. Technol. 31(5):1411-1415. Bell, J.G., C.B. Cowey and A. Youngson. 1984. Rainbow trout ( Salmo gairdneri ) liver microsomal lipid peroxidation: The effect of purified glutathione peroxidase (EC 1.11.1.9), glutathione S-transferase (EC 2.5.1.18) and other factors. Biochim. Biophys. Acta 795(1):91-99. Bell, J.G., C.B. Cowey, J.W. Adron and A.M. Shanks. 1985. Some effects of vitamin E and selenium deprivation on tissue enzyme levels and indices of tissue peroxidation in rainbow trout ( Salmo gairdneri ). Br. J. Nutr. 53:149-157. Bell, J.G., B.J.S. Pirie, J.W. Adron and C.B. Cowey. 1986a. Some effects of selenium deficiency on glutathione peroxidase (EC 1.11.1.9) activity and tissue pathology in rainbow trout ( Salmo gairdneri ). Br. J. Nutr. 55:305-311. Bell, J.G., J.W. Adron and C.B. Cowey. 1986b. Effect of selenium deficiency on hydroperoxide-stimulated release of glutathione from isolated perfused liver of rainbow trout ( Salmo gairdneri ). Br. J. Nutr. 56(2):421-428. Bell, J.G., C.B. Cowey, J.W. Adron and B.J.S. Pirie. 1987a. Some effects of selenium deficiency on enzyme activities and indices of tissue peroxidation in Atlantic salmon parr ( Salmo salar). Aquaculture 65(1):43-54. Bell, J.G., A.H. McVicar and C.B. Cowey. 1987b. Pyruvate kinase isozymes in farmed Atlantic salmon (Salmo salar): Pyruvate kinase and antioxidant parameters in pancreas disease. Aquaculture 66(1):33-42. Benemariya, H., H. Robberecht and H. Deelstra. 1991. Atomic absorption spectrometric determination of zinc, copper, and selenium in fish from Lake Tanganyika, Burundi, Africa. Sci. Total Environ. 105:73-85. Bennett, W.N. 1988. Assessment of selenium toxicity in algae using turbidostat culture. Water Res. 22(7):939-942.

### Draft November 12, 2004 Ref-7 Bennett, W.N., A.S. Brooks and M.E. Boraas. 1986. Selenium uptake and transfer in an aquatic food chain and its effects on fathead minnow ( Pimephales promelas) larvae. Arch. Environ. Contam. Toxicol. 15(5):513-517. Berg, H., M. Kibus, and N. Kautsky. 1995. Heavy metals in tropical Lake Kariba, Zimbabwe. Water Air Soil Pollut. 83(3-4):237-252. Berges, J.A. and P.J. Harrison. 1995. Relationships between nitrate reductase activity and rates of growth and nitrate incorporation under steady-state light or nitrate limitation in the marine diatom Thalassiosira pseudonana(Bacillariophyceae). J. Phycol. 31(1):85-95. Berry, M.R., L.S. Johnson, J.W. Jones, J.I. Rader, D.C. Kendall and L.S. Sheldon. 1997. Dietary characterizations in a study of human exposures in the Lower Rio Grande Valley: Part I. Foods and beverages. Environ. Int. 23(5): 675-692. Bertram, P.E. and A.S. Brooks. 1986. Kinetics of accumulation of selenium from food and water by fathead minnows (

Pimephales promelas). Water Res. 20(7):877-884. Besser, J.M., J.N. Huckins, E.E. Little and T.W. La Point. 1989. Distribution and bioaccumulation of selenium in aquatic microcosms. Environ. Pollut. 62(1):1-12. Besser, J.M., T.J. Canfield and T.W. La Point. 1993. Bioaccumulation of organic and inorganic selenium in a laboratory food chain. Environ. Toxicol. Chem. 12(1):57-72. Besser, J.M., J.N. Huckins and R.C. Clark. 1994. Separation of selenium species released from Se-exposed algae. Chemosphere 29(4):771-780. Besser, J.M., J.P. Giesy, R.W. Brown, J.M. Buell and G.A. Dawson. 1996. Selenium bioaccumulation and hazards in a fish community affected by coal fly ash effluent. Ecotoxicol. Environ. Saf. 35(1):7-15. Beyers, D.W. and C. Sodergren. 2001a. Evaluation of interspecific sensitivity to selenium exposure: larval razorback sucker versus flannelmouth sucker. Final Report to Recovery Implementation Program Project CAP-6 SE-NF. Dept. Fishery and Wildlife Biology, Colorado State Univ., Fort Collins, CO 80523. Beyers, D.W. and C. Sodergren. 2001b. Assessment of exposure of larval razorback sucker to selenium in natural waters and evaluation of laboratory-based predictions. Final Report to Recovery Implementation Program Project CAP-6 SE. Dept. Fishery and Wildlife Biology, Colorado State Univ., Fort Collins, CO 80523. Biddinger, G.R. and S.P. Gloss. 1984. The importance of trophic transfer in the bioaccumulation of chemical contaminants in aquatic ecosystems. Residue Rev. 91:103-145. Biedlingmaier, S. and A. Schmidt. 1989. Sulfate transport in normal and sulfur-deprived Chlorella fusca

. Zeitschrift Fuer Naturforschung Section C Biosciences 44(5-6):495-503. Birge, W.J. 1978. Aquatic toxicology of trace elements of coal and fly ash. In: Energy and environmental

### Draft November 12, 2004 Ref-8 stress in aquatic systems. Thorp, J.H. and J.W. Gibbons (Eds.). CONF-771114. National Technical Information Service, Springfield, VA. pp. 291-240. Birge, W.J. and J.A. Black. 1977. A continuous-flow system using fish and amphibian eggs for bioassay determinations on embryonic mortality and teratogenesis. EPA-560/5-77-002 or PB-285191. National Technical Information Service, Springfield, VA. Birge, W.J., J.E. Hudson, J.A. Black and A.G. Westerman. 1978. Embryo-larval bioassays on inorganic coal elements and in situ biomonitoring of coal-waste effluents. In: Surface mining and fish/wildlife needsin the eastern United States. Samuel, D.E., J.R. Stauffer, C.H. Hocutt and W.T. Mason (Eds.). PB298353. National Technical Information Service, Springfield, VA. pp. 97-104. Birge, W.J., J.A. Black and A.G. Westerman. 1979a. Evaluation of aquatic pollutants using fish and amphibian eggs as bioassay organisms. In: Animals as mointors of environmental pollutants. Nielson, S.W., G. Migaki and D.G. Scarrelli (Eds.). National Academy of Sciences, Washington, DC. pp. 108-118. Birge, W.J., J.A. Black, A.G. Westerman and J.E. Hudson. 1979b. The effects of mercury on reproduction of fish and amphibians. In: the biogeochemistry of mercury in the environment. Nriagu, J.O. (Ed.). Elsevier, New York, NY. pp. 629-655. Birge, W.J., J.A. Black, A.G. Westerman and J.E. Hudson. 1980. Aquatic toxicity tests on inorganic elements occurring in oil shale. In: Oil shale symposium: Sampling, analysis and quality assurance. Gale, C. (Ed.). EPA-600/9-80-022. National Technical Information Service, Springfield, VA. pp. 519-534. Birge, W.J., J.A. Black and B.A. Ramey. 1981. The reporductive toxicology of aquatic contaminants. In: Hazard assessment of chemicals: Current developments. Vol. 1. Saxena, J. and F. Fisher (Eds.). Academic Press, New York, NY. pp. 59-115. 

## Birkes, D., and Y. Dodge. 1993. Alternative methods of regression. John Wiley & Sons, New York, NY. Birkner, J.H. 1978. Selenium in aquatic organisms from seleniferous habitats. Ph.D. thesis. Colorado State University, Fort Collins, CO. Available from: University Microfilms, Ann Arbor, MI. Order No. 78- 20841. Bjerregaard, P. 1982. Accumulation of cadmium and selenium and their mutual interaction in the shore crab Carcinus maenas(L.). Aquat. Toxicol. 2:113-125. Bjerregaard, P. 1985. Effect of selenium on cadium uptake in the shore crab Carcinus maenas(L.). Aquat. Toxicol. 7:177-189. Bjerregaard, P. 1988a. Interaction between selenium and cadmium in the hemolymph of the shore crab Carcinus maenas(L.). Aquat. Toxicol. (Amsterdam) 13(1):1-12. Bjerregaard, P. 1988b. Effect of selenium on cadmium uptake in selected benthic invertebrates. Mar. Ecol. Prog. Ser. 48(1):17-28.

### Draft November 12, 2004 Ref-9 Bjerregaard, P. and L. Christensen. 1993. Accumulation of organic and inorganic mercury from food in the tissues of Carcinus maenas : Effect of waterborne selenium. Mar. Ecol. Prog. Ser. 99(3):271-281. Bjoernberg, A.A. 1989. Decontamination of mercury from Swedish "black-listed" lakes by addition of selenium. Proc. Int. Symp. Uses Selenium Tellurium, 4th. Carapella, S. C., Jr. (Ed). Selenium-Tellurium Dev. Assoc.: Darien, Conn. pp.357-360. Bjornberg, A., L. Hakanson, and K. Lundbergh. 1988. A theory on the mechanisms regulating the bioavailability of mercury in natural waters. Environ. Pollut. 49(1):53-62.Bleckmann, C.A., B. Rabe, S.J. Edgmon and D. Fillingame. 1995. Aquatic toxicity variability for freshand saltwater species in refinery wastewater effluent. Environ. Toxicol. Chem. 14(7):1219-1223. Blondin, G.A., L.M.Knobeloch, H.W. Read, and J.M. Harkin. 1988. An in vitro submitochondrial bioassay for predicting acute toxicity in fish. ASTM Spec. Tech. Publ., 1007. Aquat. Toxicol. Environ. Fate: 11th Vol., 551-63. Boisson, F. and M. Romeo. 1996. Selenium in plankton from the northwestern Mediterranean sea. Water Res. 30(11):2593-2600. Boisson, F., M. Gnassia-Barelli, J. Chiaverini and M. Romeo. 1989. Effect of selenium on the uptake of cadmium by the marine microalga Hymenomonas

(Cricosphaera) elongata . Mar. Environ. Res. 28(1-4):465-469. Boisson, F., M. Gnassia-Barelli and M.Romero. 1995. Toxicity and accumulation of selenite and selenate in the unicellular marine alga Cricosphaera elongata . Arch. Environ. Contam. Toxic. 28(4):487-493. Boisson, F., C.S. Karez, M. Henry, M. Romeo and M. Gnassia-Barelli. 1996. Ultrastructural observations on the marine coccolithophorid Cricosphaera elongatacultured in the presence of selenium or cadmium. Bull. Inst. Oceanogr. (Monaco) 0(SPEC. ISSUE 14 PART 4):239-247. Bondavalli, C., E. Croce, S. Meloni, M. Oddone and C. Triulzi. 1996. Chemical characterization of a lagoon ecosystem. The Sacca di Goro (Po River delta, Italy). Chem. Ecol. 12(4):279-286. Botsford, J.L. 1997. A simple, rapid, inexpensive assay for toxic chemicals using a bacterial indicator. Stud. Environ. Sci. 66:429-443. Botsford, J.L. J. Rivera, J. Navarez, R. Riley, T. Wright, and R. Baker. 1997. Assay for toxic chemicals using bacteria. Bull. Environ. Contam. Toxicol., 59(6):1000-1009.Bottino, N.R., C.H. Banks, K.J. Irgolick, P. Micks, A.E. Wheeler ad R.A. Zingaro. 1984. Seleniumcontaining amino acids and proteins in marine algae. Phytochem. 23:2445-2452. Bovee, E.C. 1978. Effects of heavy metals especially selenium, vanadium and zirconium on movement, growth and survival of certain aquatic life. PB-292563/4SL. National Technical Information Service, Springfield, VA.

### Draft November 12, 2004 Ref-10 Bowerman, W. W. IV, E.D. Evans, J.P Giesy, and S. Postupalsky. 1994. Using feathers to assess risk of mercury and selenium to bald eagle reproduction in the Great Lakes region. Arch. Environ. Contam. Toxicol. 27(3):294-298. Bowie, G.L., J.G. Sanders, G.F. Riedel, C.C. Gilmour, D.L. Breitburg, G.A. Cutter and D.B. Porcella. 1996. Assessing selenium cycling and accumulation in aquatic ecosystems. Water Air Soil Pollut. 90(1/2, Clean Water: Factors that Influence Its Availability, Quality and Its Use):93-104. Bowmer, T., H.A. Jenner, E. Foekema and M. van der Meer. 1994. The detection of chronic biological effects in the marine intertidal bivalve Cerastoderma edule , in model ecosystem studies with pulverized fuel ash: reproduction and histopathology. Environ. Pollut. 85(2):191-204. Boyum, K.W. 1984. The toxic effect of selenium on the zooplankton, Daphnia magnaand Daphnia pulicaria , in water and the food source ( Chamydomonas reinhardii). Ph.D. thesis. University of Wisconsin-Milwaukee, Milwaukee, WI. Available from: University Microfilms, Ann Arbor, MI. Order No. 85-09248. Boyum, K.W. and A.S. Brooks. 1983. Differential toxicity of selenium on lab cultures of Daphnia magna and field isolates ofDaphnia pulicaria . In: Proc. 26th Conf. Int. Assoc. Great Lakes Res., May 23-27, 1983, State Univ. N.Y. Oswego, Ann Arbor, MI:8. Boyum, K.W. and A.S. Brooks. 1988. The effect of selenium in water and food on Daphnia populations. Arch. Environ. Contam. Toxicol. 17(5):555-560. Braddon, S.A. 1982. Investigations into the mechanism of action of Se on Hg toxicity using a sea bass model. Abstract No. 5585. Fed. Proc. 41:1227. Braddon-Galloway, S. and J. E. Balthrop. 1985. Selenium-dependent glutathione-peroxidase isolated from black sea bass ( Centropristis striata ). Comp. Biochem. Physiol. C Comp. Pharmacol. Toxicol. 82(2):297-300. Braddon-Galloway, S. and C.R. Sumpter. 1986. A unique selenoprotein isolated from yellowfin tuna ( Thunnus albacares ) liver. Comp. Biochem. Physiol. 83C:13-17. Bradford, C.S., L. Sun and D.W. Barnes. 1994a. Basic fibroblast growth factor stimulates proliferation and suppresses melanogenesis in cell cultures derived from early zebrafish embryos. Molec. Marine Biol. Biotechnol. 3(2):78-86. Bradford, C.S., L. Sun, P. Collodi and D.W. Barnes. 1994b. Cell cultures from zebrafish embryos and adult tissues. J. Tissue Culture Methods 16(2):99-107. Brandao, J.C., H.H.L. Bohets, I.E. Van de Vyver and P.J. Dierickx. 1992. Correlation between the in vitro cytotoxicity to cultured fathead minnow fish cells and fish lethality data for 50 chemicals. Chemosphere 25(4):553-562. Brandt, A., C. Wolstrup and T.K. Nielsen. 1990. The effect of dietary dl-alpha tocopheryl acetate sodium selenite and polyunsaturated fatty acids in mink Mustela vison L. clinical chemistry and hematology. J.

### Draft November 12, 2004 Ref-11 Animal Physiol. Animal Nutr. 64(5):280-288. Brasher. A.M. and R.S. Ogle. 1993. Comparative toxicity of selenite and selenate to the amphipod Hyalella azteca . Arch. Environ. Contam. Toxicol. 24(2):182-186. Braune, B. M., R.J. Norstrom, M.P. Wong, B.T. Collins and J. Lee. 1991. Geographical Distribution of Metals in Livers of Polar Bears from the Northwest Territories Canada. Sci. Total Environ. 100:283-300. Brezina, E.R. and M.V. Arnold. 1977. Levels of heavy metals in fishes from selected Pennsylvania waters. Publication 50. Bureau of Water Quality Management, Department of Environmental Resources, Harrisburg, PA. Brieger, G., J.R. Wells and R.D. Hunter. 1992. Plant and Animal Species Composition and Heavy Metal Content in Fly Ash Ecosystems. Water Air Soil Pollut. 63(1-2):87-103. Bringmann, G. 1978. Determination of the biological toxicity of waterbound substances towards protozoa. I. Bacteriovorous flagellates (model organism: Entosiphon sulcatumStein). Z. Wasser Abwasser Forsch. 11:210-215. Bringmann, G. and R. Kuhn. 1959a. The toxic effects of waste water on aquatic bacteria, algae, and small crustaceans. Gesundh.-Ing. 80:115-120. Bringmann, G. and R. Kuhn. 1959b. Water toxicology studies with protozoans as test organisms. Gesundh.-Ing. 80:239-242. Bringmann, G. and R. Kuhn. 1976. Comparative results of the harmful effects of water pollutants on bacteria (Pseudomonas putida) and blue algae ( Microcystis aeruginosa). Gas-Wasserfach, WasserAbwasser 117:410-413. Bringmann, G. and R. Kuhn. 1977a. Limiting values for the damaging action of water pollutants to bacteria (Pseudomonas putida) and green algae ( Scenedesmus quadricauda) in the cell multiplication inhibition tests. Z. Wasser Abwasser Forsch. 10:87-98. Bringmann, G. and R. Kuhn. 1977b. Results of the damaging effect of water pollutants on Daphnia magna . Z. Wasser Abwasser Forsch. 10:161-166. Bringmann, G. and R. Kuhn. 1978a. Limiting values for the noxious effects of water pollutant material to blue algae (

Microcystis aeruginosa) and green algae ( Scenedesmus quadricauda) in cell propogation inhibition tests. Vom Wasser 50:45-60. Bringmann, G. and R. Kuhn. 1978b. Testing of substances for their toxicity threshold: Model organisms Microcystis(Diplocystis) aeruginosaand Scenedesmus quadricauda . Mitt. Int. Ver. Theor. Angew. Limnol. 21:275-284. Bringmann, G. and R. Kuhn. 1979. Comparison of toxic limiting concentrations of water contamination toward bacteria, algae and protozoa in the cell-growth inhibition test. Haustech. Bauphys. Umwelttech. 100:249-252.

### Draft November 12, 2004 Ref-12 Bringmann, G. and R. Kuhn. 1980a. Determination of the harmful biological effect of water pollutants on protozoa. II. Bacteriovorous ciliates. Z. Wasser Abwasser Forsch. 13:26-31. Bringmann, G. and R. Kuhn. 1980b. Comparison of the toxicity thresholds of water pollutants to bacteria, algae, and protozoa in the cell multiplication inhibition test. Water Res. 14:231-241. Bringmann, G. and R. Kuhn. 1981. Comparison of the effects of harmful substances on flagellates as well as ciliates and on halozoic bacteriophagous and saprozoic protozoa. Gas-Wasserfach, Wasser-Abwasser 122:308-313. Bringmann, G., R. Kuhn and A. Winter. 1980. Determination of biological damage from water pollutants to protozoa. III. Saprozoic flagellates. Z. Wasser Abwasser Forsch. 13:170-173. Brix, K.V., J.S. Volosin, W.J. Adams, R.J. Reash, R.C. Carlton and D.O. McIntyre. 2001a. Effects of sulfate on the acute toxicity of selenate to freshwater organisms. Environ. Toxicol. Chem. 5: 1037-1045. Brix, K.V., W.J. Adams, R.J. Reash, R.C. Carlton and D.O. McIntyre. 2001b. Acute toxicity of selenate on two daphnids and three gammarid amphipods. Environ. Toxicol. Chem. 16: 142-150. Brooke, L. 1987. University of Wisconsin-Superior, Superior, WI. (Memorandum to C. Stephan, U.S. EPA, Duluth, MN. July 20.) Brooke, L.T., D.J. Call, S.L. Harting, C.A. Lindberg, T.P. Markee, D.J. McCauley and S.H. Poirier. 1985. Acute toxicity of selenium(IV) and selenium(VI) to freshwater organisms. Center for Lake Superior Environmental Studies, University of Wisconsin-Superior, Superior, WI. Brooks, A.S. 1984. Selenium in the environment: An old problem with new concerns. In: Workshop proceedings: The effects of trace elements on aquatic ecosystems. EA-3329. Electric Power Research Institute, Palo Alto, CA. pp. 2-1 to 2-17. Brooks, A.S., P.E. Bertram, D.C. Szmania, D.B. Seale and M.E. Boraas. 1984. The effect of selenium on the reproductive potential of the fathead minnow. Final report on research project 1631-1. Center for Great Lakes Studies, University of Wisconsin-Milwaukee, Milwaukee, WI. Brown, T.A. and A. Shrift. 1982. Selenium: Toxicity and tolerance in higher plants. Biol. Rev. 57:59-84. Browne, C.L. and J.N. Dumont. 1979. Toxicity of selenium to developing Xenopus laevisembryos. J. Toxicol. Environ. Health 5:699-709. Browne, C. and J.N. Dumont. 1980. Cytotoxic effects of sodium selenite on tadpoles ( Xenopus laevis). Arch. Environ. Contam. Toxicol. 9:181-191. Brugmann, L. and U. Hennings. 1994. Metals in zooplankton from the Baltic Sea, 1980-84. Chem. Ecol. 9(2):87-103. Brugmann, L. and D. Lange. 1988. Trace metal studies on the starfish Asterias rubensL. from the western Baltic Sea. Chem. Ecol. 3(4):295-311.

### Draft November 12, 2004 Ref-13 Brumbaugh, W. G. and M.J. Walther. 1991. Improved Selenium Recovery from Tissue with Modified Sample Decomposition. J. Assoc. Official Anal Chemists 74(3):570-571. 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. 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 Sevices Section. Carolina Power & Light Company. September 1985. 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 Buhl, K.J. and S. J. Hamilton. 1991. Relative sensitivity of early life stages of arctic grayling coho salmon and rainbow trout to nine inorganics. Ecotoxicol. Environ. Safety 22(2):184-197. Buhl, K.J. and S.J. Hamilton. 1996. Toxicity of inorganic contaminants, individually and in environmental mixtures, to three endangered fishes (Colorado squawfish, bonytail, and razorback sucker). Arch. Environ. Contam. Toxicol. 30(1):84-92. Burger, J. 1992. Trace Element Levels In Pine Snake Hatchlings Tissue And Temporal Differences. Arch. Environ. Contam. Toxicol. 22(2):209-213. Burger, J. 1994. Heavy metals in avian eggshells: Another excretion method. J. Toxicol. Environ. Health 41(2):207-220. Burger, J. 1995. Heavy metal and selenium levels in feathers of herring gulls ( Larus argentatus ): Differences due to year, gender, and age at Captree, Long Island. Environ. Monit. Assess. 38(1):37-50. Burger, J. 1996. Heavy metal and selenium levels in feathers of Franklin's gulls in interior North America. Auk 113(2):399-407. Burger, J. 1997a. Heavy metals and selenium in herring gulls ( Larus argentatus ) nesting in colonies from eastern Long Island to Virginia. Environ. Monit. Assess. 48(3):285-296. Burger, J. 1997b. Heavy metals in the eggs and muscle of horseshoe crabs ( Limulus polyphemus ) from Delaware Bay. Environ. Monit. Assess. 46(3): 279-287. Burger, J. and M. Gochfeld. 1992a. Heavy Metal And Selenium Concentrations In Black Sk immers Rynchops niger

Gender Differences. Arch. Environ. Contam. Toxicol. 23(4):431-434. Burger, J. and M. Gochfeld. 1992b. Trace Element Distribution In Growing Feathers Additional Excretion In Feather Sheaths. Arch. of Environ. Contamin. & Toxicol. 23(1):105-108. Burger, J. and M. Gochfeld. 1993. Heavy Metal And Selenium Levels In Feathers Of Young Egrets And

### Draft November 12, 2004 Ref-14 Herons From Hong Kong And Szechuan China. Arch. Environ. Contam. Toxicol. 25(3):322-327. Burger, J. and M. Gochfeld. 1995a. Biomonitoring of heavy metals in the Pacific basin using avian feathers. Environ. Toxicol. Chem. 14(7):1233-1239. Burger, J. and M. Gochfeld. 1995b. Heavy metal and selenium concentrations in eggs of herring gulls (Larus argentatus ): Temporal differences from 1989 to 1994. Arch. Environ. Contam. Toxicol. 29(2):192-197. Burger, J. and M. Gochfeld. 1996. Heavy metal and selenium levels in Franklin's gull ( Larus pipixcan ) parents and their eggs. Arch. Environ. Contam. Toxicol. 30(4):487-491. Burger, J. and M. Gochfeld. 1997. Age differences in metals in the blood of herring ( Larus argentatus ) and Franklin's (

Larus pipixcan ) gulls. Arch. Environ. Contam. Toxicol. 33(4):436-440. Burger, J., E.A.E. Schreiber and M. Gochfeld. 1992a. Lead Cadmium Selenium And Mercury In Seabird Feathers From The Tropical Mid-Pacific. Environmental Toxicology & Chemistry 11(6):815-822. Burger, J., I.C.T. Nisbet and M. Gochfeld. 1992b. Metal Levels In Regrown Feathers Assessment Of Contamination On The Wintering And Breeding Grounds In The Same Individuals. J. Toxicol. Environ. Health 37(3):363-374. Burger, J., K. Parsons, T. Benson, T. Shukla, D. Rothstein and M. Gochfeld. 1992c. Heavy Metal And Selenium Levels In Young Cattle Egrets From Nesting Colonies In The Northeastern United States Puerto Rico And Egypt. Arch. Environ. Contam. Toxicol. 23(4):435-439. Burger, J., J.A. Rodgers, Jr. and M. Gochfeld. 1993. Heavy Metal And Selenium Levels In Endangered Wood Storks Mycteria americana From Nesting Colonies In Florida And Costa Rica. Arch. Environ. Contam. Toxicol. 24(4):417-420. Burger, J., I.C.T. Nisbet and M. Gochfeld. 1994a. Heavy metal and selenium levels in feathers of known-aged common terns ( Sterna hirundo ). Arch. Environ. Contam. Toxicol. 26(3):351-355. Burger, J., M. Pokras, R. Chafel and M. Gochfeld. 1994b. Heavy metal concentrations in feathers of common loons (

Gavia immer

) in the Northeastern United States and age differences in mercury levels. Environ. Monitor. Assess. 30(1):1-7.Burnham, K. P., and D. R. Anderson. 2002. Model selection and inference: a practical informationtheoretic approach. Springer, New York, NY. Burton, D.T., L.W. Hall, Jr., R.J. Klauda and S.L. Margrey. 1983. Effects of treated bleached kraft mill effluent on eggs and prolarvae of striped bass ( Morone saxatilis ). Water Resour. Bull. 19:869-879. Burton, G.A., Jr. and B.L. Stemmer. 1988. Evaluation of surrogate tests in toxicant impact assessments. Toxic. Assess. 3(3):255-69. Burton, G.A., Jr., A. Drotar, J.M. Lazorchak, L.L. Bahls. 1987a. Relationship of microbial activity and

### Draft November 12, 2004 Ref-15 Cerio daphnia responses to mining impacts on the Clark Fork River, Montana. Arch. Environ. Contam. Toxicol. 16(5):523-30. Burton, G.A. Jr., T.H. Giddings, P. Debrine and R. Fall. 1987b. High incidence of selenite-resistant bacteria from a site polluted with selenium. Appl. Environ. Microbiol. 53(1):185-188. Burton, G.A., Jr., D. Nimmo, D. Murphey and F. Payne. 1987c. Stream profile determinations using microbial activity assays and Ceriodaphnia . Environ. Toxicol. Chem. 6(7):505-13. Burton, J.D., W.A. Maher, C.I. Measures and P.J. Statham. 1980. Aspects of the distribution and chemical form of selenium and arsenic in ocean waters and marine organisms. Thalassia Jugosl. 16:155-164. Burton, W.H. and A.E. Pinkney. 1994. Yellow perch larval survival in the Zekiah Swamp watershed (Wicomico River, Maryland) relative to the potential effects of a coal ash storage facility. Water Air Soil Pollut.72(1-4):235-249. Butler, G.W. and P.J. Peterson. 1967. Uptake and metabolism of inorganic Se-75 by Spirodela olingorrhiza . Aust. J. Biol. Sci. 20:77-86. Byl, T.D., H.D. Sutton and S. J. Klaine. 1994. Evaluation of peroxidase as a biochemical indicator of toxic chemical exposure in the aquatic plant Hydrilla verticillata , Royle. Environ. Toxicol. Chem. 13(3):509-515. Byrne, C.J. and L.R. DeLeon. 1986. Trace metal residues in biota and sediments from Lake Pontchartrain, Louisiana. Bull. Environ. Contam. Toxicol. 37(1):151-8 Byrne, C., R. Balasubramanian, E.B. Overton and T.F. Albert. 1985. Concentrations of trace metals in the bowhead whale. Mar. Pollut. Bull. 16(12):497-8. Cade, B., and J. D. Richards. 1996. Permutation tests for least absolute deviation regression. Biometrics. 52:886-902. Cade, B. S., J. W. Terrel, and R. L. Schroeder. 1999. Estimating effects of limiting factors with regression quantiles. Ecology. 80:311-323. Cade BS, Noon BR. 2003. A gentle introduction to quantile regression for ecologists. Front Ecol Env 1:412-420. Caffrey, P.B. 1989. The effects of zinc deprivation on selenium requirements in daphnids (Crustacea). 169 pp. Available from Univ. Microfilms Int., Order No. DA9008884 From: Diss. Abstr. Int. B 1990, 50(11):4959. Call, D.J., L.T. Brooke, N. Ahmad and J.E. Richter. 1983. Toxicity and metabolism studies with EPA (Environmental Protection Agency) priority pollutants and related chemicals in freshwater organisms. PB83-263665 or EPA-600/3-83-095. National Technical Information Service, Springfield, VA. Callahan, M.A., M.W. Slimak, N.W. Gabel, I.P. May, C.F. Fowler, J.R. Freed, P. Jennings, R.L. Durfee,

### Draft November 12, 2004 Ref-16 F.C. Whitmore, B. Maestri, W.R. Mabey, B.R. Holt and C. Gould. 1979. Water-related environmental fate of 129 priority pollutants. Vol. I. EPA-440/4-79-029a. National Technical Information Service, Springfield, VA. pp. 16-1 to 16-13. Cantillo, A.Y., G.C. Lauenstein, and T.P. O'connor. 1997. Mollusc and sediment contaminant levels and trends in south Florida coastal waters. Mar. Pollut. Bull. 34(7):511-521. Canton, S.P. 1999. Acute aquatic life criteria for selenium. Environ. Toxicol. Chem. 18:1425-1432.Canton, S.P. and W.D. VanDerveer. 1997. Selenium toxicity to aquatic life: an argument for sedimentbased water quality criteria. Environ. Toxicol. Chem. 16(6):1255-1259. Capar, S.G. and N.J. Yess. 1996. U.S. food and drug administration survey of cadmium, lead and other elements in clams and oysters. Food Addit. Contam. 13(5):553-560. Capelli, R., V. Minganti and M. Bernhard. 1987. Total mercury, organic mercury, copper, manganese, selenium, and zinc inSarda sarda from the Gulf of Genoa. Sci. Total Environ. 63(0):83-100. Capelli, R., V. Minganti, F. Fiorention and R. DePellegrini. 1991. Mercury and Selenium in Adamussium colbecki and Pagothenia bernacchii from the Ross Sea Antarctica Collected During Italian Expedition 1988-89. Ann. Chim. 81(7-8):357-370. Cappon, C.J. 1984. Content and chemical form of mercury and selenium in Lake Ontario USA salmon and trout. J. Great Lakes Res. 10:429-434. Cappon, C.J. and J.C. Smith. 1981. Mercury and selenium content and chemical form in fish muscle. Arch. Environ. Contam. Toxicol. 10:305-319. Cappon, C.J. and J.C. Smith. 1982a. Chemical form and distribution of mercury and selenium in edible seafood. J. Anal. Toxicol. 6:10-21. Cappon, C.J. and J.C. Smith. 1982b. Chemical form and distribution of mercury and selenium in canned tuna. J. Appl. Toxicol. 2:181-189. Cardellicchio, N. 1995. Persistent contaminants in dolphins: An indication of chemical pollution in the Mediterranean Sea. Water Sci. Technol. 32(9-10):331-340. Cardin, J.A. 1986. U.S. EPA, Narragansett, RI. (Memorandum to D.J. Hansen, U.S. EPA, Narrangansett, RI.) Cardwell, R.D., D.G. Foreman, T.R. Payne and D.J. Wilbur. 1976a. Acute toxicity of selenium dioxide to freshwater fishes. Arch. Environ. Contam. Toxicol. 4:129-144. Cardwell, R.D., D.G. Foreman, T.R. Payne and D.J. Wilbur. 1976b. Acute toxicity of selected toxicants to six species of fish. PB-252488 or EPA-600/3-76-008. National Technical Information Service, Springfield, VA.

### Draft November 12, 2004 Ref-17 Carell, B., S. Forberg, E. Grundelius, L. Henrikson, A. Johnels, U. Lindh, H. Mutvei, M. Olsson, K. Svaerdstroem and T. Westermark. 1987. Can mussel shells reveal environmental history?. Ambio 16(1):2-10. Carter, L.F. and S.D. Porter. 1997. Trace-element accumulation by Hygrohypnum ochraceum in the upper Rio Grande Basin, Colorado and New Mexico, USA. Environ. Toxicol. Chem. 16(12):2521-2528. Caurant, F., J.C. Amiard, C. Amiard-Triquet and P.G. Sauriau. 1994. Ecological and biological factors controlling the concentrations of trace elements (As, Cd, Cu, Hg, Se, Zn) in delphinids Globicephala melas from the North Atlantic Ocean. Mar. Ecol.: Prog. Ser. 103(3):207-219. Caurant, F., M. Navarro, and J.C. Amiard. 1996. Mercury in pilot whales: Possible limits to the detoxification process. Sci. Total Environ. 186(1-2):95-104. Chandy, J. P. and B. Patel. 1985. Do selenium and glutathione detoxify mercury in marine invertebrates? Effects on lysosomal response in the tropical blood clam Anadara granosa . Dis. Aquat. Organisms 1(1):39-48. Chapman, D.C. 1992. Failure of gas bladder inflation in striped bass: effect on selenium toxicity. Arch. Environ. Contam. Toxicol. 22(3):296-299. Chapman, W.H., H.L. Fisher and M.W. Pratt. 1986. Concentration factors of chemical elements in edible aquatic organisms. UCRL-50564. National Technical Information Service, Springfield, VA. Chau, Y.K. and J.P. Riley. 1965. The determination of selenium in sea water, silicates, and marine organisms. Anal. Chim. Acta 33:36-49. Chau, Y.K., P.T.S. Wong, B.A. Silverberg, P.L. Luxon and G.A. Bengert. 1976. Methylation of selenium in the aquatic environment. Science 192:1130-1131. Chen, C., Y. Liu, J. Zhou, H. Xu and S. Qu. 1997. Microcalorimetric study of the toxic effect of selenium on the mitochondrial metabolism of Cyprinus carpoliver. Biol. Trace Element Rev. 60:115-122. Cheng, L. L., P.R. Bowser and J.M. Spitsbergen. 1993. Development Of cell cultures derived from lake trout liver and kidney in a hormone-supplemented serum-rediced medium. J. Aquat. Animal Health 5(2):119-126. Cherry, D.S. and R.K. Guthrie. 1978. Mode of elemental dissipation from ash basin effluent. Water Air Soil Pollut. 9:403-412. Cherry, D.S., R.K. Guthrie, J.H. Rodgers, Jr., J. Cairns, Jr. and K.L. Dickson. 1976. Responses of mosquitofish (

Gambusia affinis ) to ash effluent and thermal stress. Trans. Am. Fish. Soc. 105:686-694. Cherry, D.S., R.K. Guthrie, F.F. Sherberger and S.R. Larrick. 1979a. The influence of coal ash and thermal discharges upon the distribution and bioaccumulation of aquatic invertebrates. Hydrobiol. 62:257- 267.

### Draft November 12, 2004 Ref-18 Cherry, D.S., S.R. Larrick, R.K. Guthrie, E.M. Davis and F.F. Sherberger. 1979b. Recovery of invertebrate and vertebrate populations in a coal ash stressed drainage system. J. Fish. Res. Board Can. 36:1089-1096. Cherry, D.S., R.K. Guthrie, E.M. Davis and R.S. Harvey. 1984. Coal ash basin effects (particulates, metals, acidic pH) upon aquatic biota: An eight year evaluation. Water Resour. Bull. 20:535-544. Cherry, D.S., J.H. Van Hassel, P.H. Ribbe and J. Cairns, Jr. 1987. Factors influencing acute toxicity of coal ash to rainbow trout ( Salmo gairdneri ) and bluegill sunfish ( Lepomis macrochirus). Water Resour. Bull. 23(2):293-306. Chiang, L., B.D. James, R.J. Magee. 1994. Determination of selenium in biological and environmental samples by adsorptive stripping voltammetry. Malays. J. Sci. Ser. B, 15(1 & 2):31-34. Chidambaram, N. and C.A. Sastry. 1991a. Toxicity and bioaccumulation of selenate in the teleost fish, Oreochromis mossambicus(Peters). Indian J. Environ. Prot. 11(7):496-501. Chidambaram, N. and C.A. Sastry. 1991b. Some aspects of selenium accumulation in a freshwater teleost fish, Oreochromis mossambicus(Peters). Indian J. Environ. Prot. 11(10):761-770. Chou, C.L. and J.F. Uthe. 1991. Effect of starvation on trace metal levels in blue mussels ( Mytilus edulis ). Bull. Environ. Contam. Toxicol. 46(3):473-478. Christensen, G.M. and J.H. Tucker. 1976. Effects of selected water toxicants on the in vitro activity of fish carbonic anhydrase. Chem.-Biol. Interact. 13:181-192. Chvojka, R. 1988. Mercury and selenium in axial white muscle of yellowtail kingfish from Sydney, Australia. Mar. Pollut. Bull. 19(5):210-213. Chvojka, R., R.J. Williams and S. Frederickson. 1990. Methyl Mercury Total Mercury and Selenium in Snapper from Two Areas of the New South Wales Coast Australia. Mar. Pollut. Bull. 21(12):570-573. Cieminski, K. L. and L.D. Flake. 1995. Invertebrate fauna of wastewater ponds in southeastern Idaho. Great Basin Naturalist 55(2):105-116. Clark, D.R. Jr., P.A. Ogasawara, G.J. Smith and H.M. Ohlendorf. 1989. Selenium accumulation by raccoons exposed to irrigation drainwater at Kesterson National Wildlife Refuge California USA 1986. Arch. Environ. Contam. Toxicol. 18(6):787-794. Cleveland, L., E.E. Little, D.R. Buckler and R.H. Wiedmeyer. 1993. Toxicity and bioaccumulation of waterborne and dietary selenium in juvenile bluegill ( Lepomis macrochirus). Aquat. Toxicol. (Amsterdam) 27(3-4):265-279. Clifford, P.J. and P.J. Harrison. 1988. Use of carbon-14 uptake rates to evaluate the selenium nutrition of a marine phytoplankter, Thalassiosira pseudonana (Hustedt) Hasle et Heimdal. J. Exp. Mar. Biol. Ecol. 124(2):87-96.

### Draft November 12, 2004 Ref-19 Clifford, D., S. Subrammian and T.J. Sorg. 1986. Removing dissolved inorganic contaminants from water. Environmental Science of Technology . 20:1072- . Collins, C. T. 1992. Metals in eggs of the California least tern in Southern California. Bull. South. Calif. Acad. Sci. 91(2):49-54. Combs, G.F. Jr., C. Garbisu, B.C. Yee, A. Yee, D.E. Carlson, N.R. Smith, A.C. Magyarosy, T. Leighton and B.B. Buchanan. 1996. Bioavailability of selenium accumulated by selenite-reducing bacteria. Biol. Trace Element Res. 52(3):209-225. Congiu, A.M., S. Casu and G. Ugazio. 1989. Toxicity of selenium and mercury on the planarian Dugesia gonocephala . Res. Comm. Chem. Pathol. Pharmacol. 66(1):87-96. Cooke, T.D., and K.W. Bruland. 1987. Aquatic Chemistry of Selenium: Evidence of Biomethylation. Environmental Science of Technology . 21:1214- . Cooke, T.D. and C.-C. Lee. 1993. Toxicity Identification Evaluations (TIE) in San Francisco Bay area urban storm water runoff. Proc. - Water Environ. Fed. Annu. Conf. Expo., 66th, Volume 7. Water Environ. Fed.: Alexandria, Va. pp. 369-378. Cooney, J. D., G.M. DeGraeve, E.L. Moore, W.D. Palmer and T.L. Pollock. 1992. Effects of Food and Water Quality on Culturing of CerioDaphnia dubia . Environ. Toxicol. & Chem. 11(6):823-837. Cooper, W.C., K.G. Bennett and F.C. Croxton. 1974. The history, occurrence, and properties of selenium. In: Selenium. Zingaro, R.A. and W.C. Cooper (Eds.). Van Nostrand Reinhold Company, New York, NY. pp. 1-30. Cosson, R.P., J.C. Amiard, and C. Triquet-Amiard. 1988. Trace elements in little egrets and flamingos of Camargue, France. Ecotoxicol. Environ. Safety 15(1):107-116 Cossu, C., A. Doyette, M.C. Jacquin, M. Babut, A. Exinger and P. Vasseur. 1997. Glutathione reductase, selenium-dependent glutathione peroxidase, glutahione levels, and lipid peroxidation in freshwater bivalves, Unio tumidus , as biomarkers of aquatic contamination in field studies. Ecotox. Environ. Saf. 38:122-131. Cotton, F.A., and G. Wilkinson. Advanced Inorganic Chemistry, 5th Ed. Wiley, New York, 1988. Coughlan, D.J. and J.S. Velte. 1989. Dietary toxicity of selenium-contaminated red shiners to striped bass. Trans. Am. Fish. Soc. 118(4):400-408. Courtney, A.J., D. J. Die and M.J. Holmes. 1994. Discriminating populations of the eastern king prawn, Penaeus plebejus , from different estuaries using ICP-MS trace element analysis. At. Spectrosc. 15(1):1-6. Cowgill, U.M. 1987. Critical analysis of factors affecting the sensitivity of zooplankton and the reproducibility of toxicity test results. Water Res. 21(12):1453-1462. Cowgill, U.M. and D.P. Milazzo. 1989. The culturing and testing of two species of duckweed. ASTM Spec. Tech. Publ. 1027 (Aquat. Toxicol. Hazard Assess.: 12th Vol.):379-391.

### Draft November 12, 2004 Ref-20 Cowgill, U.M., H.W. Emmel and I.T. Takahashi. 1985. Inorganic chemical composition of trout food pellets and alfalfa used to sustain Daphnia magnaStraus. Bull. Environ. Contam. Toxicol. 34:890-896. Cowgill, U.M., H.W. Emmel, D.L. Hopkin, S.L. Applegath and I.T. Takahashi. 1986. The influence of water on reproductive success and chemical composition of laboratory reared populations of Daphnia magna . Water Res. 20:317-323. Coyle, J.J., D.R. Buckler, C.G. Ingesoll, J.F. Fairchild and T.W. May. 1993. Effect of dietary selenium on the reproductive success of bluegills Lepomis macrochirus . Environ. Toxicol. Chem. 12(3):551-565. Crane, M., T. Flower, D. Holmes and S. Watson. 1992. The toxicity of selenium in experimental freshwater ponds. Arch. Environ. Contam. Toxicol. 23(4):440-452. Crock, J. G., R.C. Severson and L.P. Gough. 1992. Determining Baselines and Variability of Elements in Plants and Soils near the Kenai National Wildlife Refuge Alaska. Water Air Soil Pollut. 63(3-4):253-271. Cruwys, E., K. Robinson and N.R. Davis. 1994. Microprobe analysis of trace metals in seal teeth from Svalbard, Greenland, and South Georgia. Polar Rec. 30(172):49-52. Cumbie, P.M. and S.L. Van Horn. 1978. Selenium accumulation associated with fish mortality and reproductive failure. Proc. Annu. Conf. Southeast. Assoc. Fish Wildl. Agencies 32:612-624. Currey, N.A., W.I. Benko, B.T. Yaru and R. Kabi. 1992. Determination of heavy metals, arsenic and selenium in barramundi (

Lates calcarifer) from Lake Murray, Papua New Guinea. Sci. Total Environ. 125:305-320. Cushman, R.M., S.G. Hildebrand, R.H. Strand and R.M. Anderson. 1977. The toxicity of 35 trace elements in coal to freshwater biota: A data base with automated retrieval capabilities. ORNL/TM-5793. National Technical Information Service, Springfield, VA. Custer, T.W. and W.L. Hohman. 1994. Trace elements in canvasbacks ( Aythya valisineria) wintering in Louisiana, USA, 1987-1988. Environ. Pollut. 84(3):253-259. Custer, T.W. and C.A. Mitchell. 1991. Contaminant Exposure of Willets Feeding in Agricultural Drainages Of the Lower Rio Grande Valley of South Texas USA. Environ. Monitor. Assess. 16(2):189-200. Custer, T.W. and C.A. Mitchell. 1993. Trace Elements and Organochlorines in the shoalgrass community of the lower Laguna Madre Texas. Environ. Monitor. Assess. 25(3):235-246. Custer, T.W., R.K. Hines, M.J. Melancon, D.J. Hoffman, J.K. Wickliffe, J.W. Bickham, J.W. Martin and D.S. Henshel. 1997. Contaminant concentrations and biomarker response in great blue heron eggs from 10 colonies on the Upper Mississippi River, USA. Environ. Toxicol. Chem. 16(2):260-271. Cutter, G.A. 1985. Determination of selenium speciation in biogenic particles and sediments. Anal. Chem. 57(14):2951-2955.

### Draft November 12, 2004 Ref-21 Cutter, G.A. 1989. The Estuarine Behavior of Selenium in San Francisco Bay. Estuarine, Coastal and Shelf Science. 28:13- . Cutter, G.A. 1986. Speciation of Selenium and Arsenic in Natural Waters and Sediments, Volume 1. Report Ea-4641. Electric Power Research Institute, Palo Alto, CA Cutter, G.A. and K.W. Bruland. 1984. The marine biogeochemisty of selenium: A re-evaluation. Limnol. Oceanogr. 29:1179-1192. Cutter, G.A., and L.S. Cutter. 1995. Behavior of Dissolved Antimonly. Arsenic, and Selenium in the Atlantic Ocean. Marine Chemistry . 49:295- . Cuvin, M.L.A. and R.W. Furness. 1988. Uptake and elimination of inorganic mercury and selenium by minnows Phoxinus phoxinus . Aquat. Toxicol. (Amsterdam) 13(3):205-216. Dabbert, C.B. and K.C. Powell. 1993. Serum enzymes as indicators of capture myopathy in mallards Anas platryhynchos . J. Wildl. Dis. 29(2):304-309. Dabeka, R.W. and A.D. McKenzie. 1991. Graphite-furnace atomic absorption spectrometric determination of selenium in foods after sequential wet digestion with nitric acid, dry ashing and coprecipitation with palladium. Can. J. Appl. Spectrosc. 36(5):123-126. Davies, A.G. 1978. Pollution studies with marine plankton. Part II. Heavy metals. Adv. Mar. Biol. 15:381- 508. Davies, I.M. and R. Russell. 1988. The influence of dissolved selenium compounds on the accumulation of inorganic and methylated mercury compounds from solution by the mussel Mytilus edulisand the plaice Pleuronectes platessa. Sci. Total Environ. 168(0):197-206. Davoren, W.T. 1986. Selenium and San Francisco Bay. In: Selenium and agricultural drainage: Implications for San Francisco Bay and the California environment. The Bay Institute of San Francisco, Tiburon, CA. pp. 150-162. Deaker, M. and W. Maher. 1997. Low-volume microwave digestion for the determination of selenium in marine biological tissues by graphite furnace atomic absorption spectroscopy. Anal. Chim. Acta, 350(3):287-294. De Jong, L.E.D.D. 1965. Tolerance of Chlorella vulgarisfor metallic and non-metallic ions. Antonie Leeuwenhoek J. Microbiol. Serol. 31:301-313. de Peyster, A., R. Donohoe, D.J. Slymen, J.R. Froines, A.W. Olivieri and D.M. Eisenberg. 1993. Aquatic biomonitoring of reclaimed water for potable use: the San Diego health effects study. J. Toxicol. Environ. Health, 39(1):121-142. Deelstra, H., P. Van Dael, R. Van Cauwenbergh and H. Robberecht. 1989. Interaction of heavy metals on the availability of selenium compounds to Artemia salina . Spec. Publ. - R. Soc. Chem. 72(Nutr. Availability: Chem. Biol. Aspects):284-286.

### Draft November 12, 2004 Ref-22 Demon, A., M. DeBruin and H.T. Wolterbeek. 1988. The influence of pH on trace element uptake by an alga (Scenedesmus pannonicus ssp. Berlin) and fungus (

Aureobasidium pullulans ). Environ. Monitor. Assess. 10(2):165-174. DeQuiroga, G.B., M. Lopez-Torres, and P. Gil. 1989. Hyperoxia decreases lung size of amphibian tadpoles without changing GSH-peroxidases or tissue peroxidation. Comp. Biochem. Physiol. A Comp. Physiol. 92(4):581-588. Devillers, J., A. Elmouaffek, D. Zakarya, and M. Chastrette. 1988. Comparison of ecotoxicological data by means of an approach combining cluster and correspondence factor analyses. Chemosphere 17(4):633-46. Dickman, M. and G. Rygiel. 1996. Chironomid larval deformity frequencies, mortality, and diversity in heavy-metal contaminated sediments of a Canadian riverine wetland. Environ. Int. 22(6):693-703. Dierenfeld, E.S., C.D. Sheppard, J. Langenberg, C. Mirande, J. Spratt and F.J. Dein. 1993. Vitamin E In Cranes Reference Ranges And Nutrient Interactions. J. Wildl. Dis. 29(1):98-102. Dierickx, P.J. 1993. Correlation between the reduction of protein content in cultured FHM fish cells and fish lethality data. Toxicol. in Vitro 7(4):527-530. Dietrich, C.P., H.B. Nader, L. Toma, P. DeAzambuya and E.S. Garcia. 1987. A relationship between the inhibition of heparan sulfate and chondroitin sulfate synthesis and the inhibition of molting by selenate in the hemipteran Rhodnius prolixus . Biochem. Biophys. Res. Comm. 146(2):652-658. Dietz, R., E.W. Born, C.T. Agger and C.O. Nielsen. 1995. Zinc, cadmium, mercury and selenium in polar bears ( Ursus maritimus ) from Central East Greenland. Polar Biol. 15(3):175-185. Dietz, R., F. Riget and P. Johansen. 1996. Lead, cadmium, mercury and selenium in Greenland marine animals. Sci. Total Environ. 186(1-2):67-93. DiIlio, C., G. DelBoccio, M. Miranda, A. Manilla, O. Zarivi and G. Federici. 1986. Glutathione peroxidases and glutathione reductase activities during Bufo bufo development. Comp. Biochem. Physiol. C Comp. Pharmacol. Toxicol. 83(1):9-12. Ding, L., et al. 1988. Study of inhibitive effect of sodium selenite on hepatocarcinogenesis in ducks. Acta Academiae Medicinae Sinicae 10(2):100-103. Dobbs, M.G., D.S. Cherry, J. Cairns Jr. 1996. Toxicity and bioaccumulation of selenium to a three-trophic level food chain. Environ. Toxicol. Chem. 15(3):340-347. Doherty, F.G., D.W. Evans, E.F. Neuhauser. 1993. An assessment of total and leachable contaminants in zebra mussels (

Dreissena polymorpha) from Lake Erie. Ecotoxicol. Environ. Saf. 25(3):328-340. Doran, J.W. 1982. Microorganisms and the biological cycling of selenium. Adv. Microb. Ecol. 6:1-32. Doucette, G.J., N.M. Price and P.J. Harrison. 1987. Effects of selenium deficiency on the morphology and

### Draft November 12, 2004 Ref-23 ultrastructure of the coastal marine diatom Thalassiosira pseudonana (Bacillariophyceae). J. Phycol. 23(1):9-17. Doyotte, A., C. Cossu, M.C. Jacquin, M. Babut, and P. Vasseur. 1997. Antioxidant enzymes, glutathione and lipid peroxidation as relevant biomarkers of experimental or field exposure in the gills and the digestive gland of the freshwater bivalve Unio tumidus . Aquat. Toxicol. 39(2):93-110. Draper, N.R. and H. Smith. 1981. Applied Regression Analysis. Second Edition.Wiley, New York. 709. Drndarski, N., D. Stojic, M. Zupancic and S. Cupic. 1990. Determination of partition coefficients of metals in the Sava River environment. J. Radioanal. Nucl. Chem. 140(2):341-348. Drotar, A., L.R. Fall, E.A. Mischalanie, J.E. Tavernier, and R. Fall. 1987. Enzymatic methylation of sulfide, selenide, and organic thiols by Tetrahymena thermophila . Appl. Environ. Microbiol. 53(9):2111-2118. Dubois, W. and G.V. Callard. 1993. Culture of intact sertoli-germ cell units and isolated sertoli cells from squalus testis II: Stimulatory effects of insulin and IGF-I on DNA synthesis in premeiotic stages. J. Exp. Zool. 267(2):233-244. Dubowy, P.J. 1989. Effects of diet on selenium bioaccumulation in marsh birds. J. Wildl. Manag. 53(3):776-781. Dunbar, A.M., J.M. Lazorchak and W.T. Waller. 1983. Acute and chronic toxicity of sodium selenate to Daphnia magnaStraus. Environ. Toxicol. Chem. 2:239-244. Duncan, D.A. and J.F. Klaverkamp. 1983. Tolerance and resistance to cadmium in white suckers ( Catastomus commersoni) previously exposed to cadmium, mercury, zinc, or selenium. Can. J. Fish. Aquat. Sci. 40:128-138. Ebringer, L., J. Dobias, J. Krajcovic, J. Polonyi, L. Krizkova and N. Lahitova. 1996. Antimutagens reduce ofloxacin-induced bleaching in Euglena gracilis . Mutat. Res. 359(2):85-93. Eisler, R. 1985. Selenium hazards to fish, wildlife, and invertebrates: A synoptic review. Contaminant Hazard Reviews. Report No. 5. Biological Report 85(1.5). U.S. Fish and Wildlife Service, Laurel, MD. Elendt, B. P. 1990. Selenium Deficiency in Crustacea, an Ultrastructural Approach to Antennal Damage in Daphnia magnaStraus. Protoplasma 154(1):25-33. Elendt, B. P. and W.R. Bias. 1990. Trace Nutrient Deficiency inDaphnia magnaCultured in Standard Medium for Toxicity Testing Effects of the Optimization of Culture Conditions on Life History Parameters of Daphnia magna . Water Res. 24(9):1157-1168. Elliott, J.E. and A.M. Scheuhammer. 1997. Heavy metal and metallothionein concentrations in seabirds from the Pacific Coast of Canada. Mar. Pollut. Bull. 34(10):794-801. Ellis, M.M. 1937. Detection and measurement of stream pollution. Bulletin No. 22. Bureau of Fisheries,

### Draft November 12, 2004 Ref-24 U.S. Department of Commerce, Washington, DC. Ellis, M.M., H.L. Motley, M.D. Ellis and R.O. Jones. 1937. Selenium poisoning in fishes. Proc. Soc. Exp. Biol. Med. 36:519-522. Enberg, A. and L. Wu. 1995. Selenium assimilation and differential response to elevated sulfate and chloride salt concentrations in two saltgrass ecotypes. Ecotoxicol. Environ. Saf. 32(2):171-178. Engberg, R. M. and C.F. Borsting. 1994. Inclusion of oxidized fish oil in mink diets. 2. The influence on performance and health considering histopathological, clinical-chemical, and haematological indices. J. Animal Physiol. Animal Nutr. 72(2-3):146-157. Engberg, R. M., K. Jakobsen, C.F. Borsting and H. Gjern. 1993. On The Utilization Retention And Status Of Vitamin E In Mink Mustela vison Under Dietary Oxidative Stress. J. Animal Physiol. Animal Nutr. 69(2-3):66-78. Eriksson, C. and C. Forsberg. 1992. Nutrient interactions and phytoplankton growth during the spring bloom period in Lake Erken, Sweden. Int. Rev. Gesamten Hydrobiol. 77(4):517-551. Eriksson, C. and C. Pedros-Alio. 1990. Selenium as a nutrient for freshwater bacterioplankton and its interactions with phosphorus. Can. J. Microbiol. 36(7):475-483. Eriksson, M.O.G., L. Henrikson and H.G. Oscarson. 1989. Metal contents in liver tissues of non-fledged goldeneye,Bucephala clangula , ducklings: a comparison between samples from acidic, circumneutral, and limed lakes in South Sweden. Arch. Environ. Contam. Toxicol. 18(1-2):255-60. Eun, J.B., J.O. Hearnsberger and J.M. Kim. 1993. Antioxidants, activators, and inhibitors affect the enzymic lipid peroxidation system of catfish muscle microsomes. J. Food Sci. 58(1):71-74. Evans, D.W., D.K. Dodoo and P.J. Hanson. 1993. Trace element concentrations in fish livers: implications of variations with fish size in pollution monitoring. Mar. Pollut. Bull. 26(6):329-334. Ewan, R.C. 1979. Toxicology and adverse effects of mineral imbalance with emphasis on selenium and other minerals. In: Toxicity of heavy metals in the environment. Part 1. Oehme, F.W. (Ed.). Marcel Dekker, New York, NY. pp. 445-489. Fairbrother, A., M. Fix, T. O ’ Hara and C.A. Ribic. 1994. Impairment of growth and immune function of avocet chicks from sites with elevated selenium arsenic and boron. J. of Wildl. Dis. 30(2):222-233. Fan, T.W.M., A.N. Lane, and R.M. Higashi. 1997. Selenium biotransformations by a euryhaline microalga isolated from a saline evaporation pond. Environ. Sci. Tech. 31(2):569-576. Fan, T.W.M., S.J. Teh, D.E. Hinton, and R.M. Higashi. 2002. Selenium Biotransformations into Proteinaceous Forms by Foodweb Organisms of Selenium-Laden Drainage Waters in California. Aquatic Toxicology . 57:65- . Fava, J.A., J.J. Gift, A.F. Maciorowski, W.L. McCulloch and H.J. Reisinger II. 1985a. Comparative

### Draft November 12, 2004 Ref-25 toxicity of whole and liquid phase sewage sludges to marine organisms. In: Aquatic toxicology and hazard assessment: Seventh symposium. Cardwell, R.D., R. Purdy and R.C. Bahner (Eds.). ASTM STP 854. American Society for Testing and Materials, Philadelphia, PA. pp. 229-252. Fava, J.A., W.L. McCulloch, J.J. Gift, H.J. Reisinger, S.E. Storms, A.F. Maciorowski, J.E. Edinger, and E. Buchak. 1985b. A multidisciplinary approach to the assessment of ocean sewage sludge disposal. Environ. Toxicol. Chem. 4(6):831-840. Faust, S.D., and O.M. Aly. 1981. Chemistry of Natural Waters . Ann Arbor Science, Ann Arbor, MI. pp. 359-371. Felton, S.P., W. Ji and S.B Mathews. 1990. Selenium Concentrations in Coho Salmon Outmigrant Smolts and Returning Adults a Comparison of Wild Versus Hatchery-reared Fish. Dis. Aquat. Organ. 9(2):157-161. Felton, S.P., R. Grace and M. Landolt. 1994. Significantly higher levels of zinc and copper found in wild compared to hatchery-reared coho salmon smolts Oncorhynchus kisutch . Dis. Aquat. Organ. 18(3):233-236. Felton S.P., M.L. Landolt, R. Grace, and A. Palmisano. 1996. Effects of selenium dietary enhancement on hatchery-reared coho salmon, Oncorhynchus kisutch (Walbaum), when compared with wild coho: Hepatic enzymes and seawater adaptation evaluated. Aquac. Res. 27(2):135-142. Feroci, G., A. Fini, R. Badiello, and A. Breccia. 1997. Interaction between selenium derivatives and heavy metal ions: Cu-2+ and Pb-2+. Microchem. J. 57(3):379-388. Finger, S.E. and J.S. Bulak. 1988. Toxicity of water from three South Carolina rivers to larval striped bass. Trans. Am. Fish. Soc. 117(6):521-8. Finley, K.A. 1985. Observations of bluegills fed selenium-contaminated Hexagenia

nymphs collected from Belews Lake, North Carolina. Bull. Environ. Contam. Toxicol. 35:816-825. Fishbein, L. 1984. Overview of analysis of carcinogenic and/or mutagenic metals in biological and environmental samples. I. Arsenic, beryllium, cadmium, chromium and selenium. Int. J. Environ. Anal. Chem. 17:113-170. Fisher, N.S. and J.R. Reinfelder. 1991. Assimilation of selenium in the marine copepod Acartia tonsa studied with a radiotracer ratio method. Mar. Ecol. Prog. Ser. 70(2):157-164. Fisher, N.S. and M. Wente. 1993. The release of trace elements by dying marine phytoplankton. Deep-Sea Res., Part I, 40(4):671-694. Fitzsimons, J.D., S. Huestis and B. Williston. 1995. Occurrence of a swim-up syndrome in Lake Ontario lake trout in relation to contaminants and cultural practices. J. Great Lakes Res. 21(Suppl. 1, International Conference on Restoration of Lake Trout in the Laurentian Great Lakes, 1994):277-285. Fjeld, E. and S. Rognerud. 1993. Use of path analysis to investigate mercury accumulation in brown trout

### Draft November 12, 2004 Ref-26 (Salmo trutta ) in Norway and the influence of environmental factors. Can. J. Fish. Aquat. Sci. 50(6):1158-1167. Fjolstad, M. and A. L. Heyeraas. 1985. Muscular and myocardial degeneration in cultured Atlantic salmon, Salmo salar , suffering from "Hitra disease". J. Fish Dis. 8(4):367-372. Fletcher, C.A., J.M. Bubb and J.N. Lester. 1994. Magnitude and distribution of anthropogenic contaminants in salt marsh sediments of the Essex coast, UK. II. Selected metals and metalloids. Sci. Total Environ. 155(1):47-59. Flury, M., W.T. Frakenberger and W.A. Jury. 1997. Long-term depletion of selenium from Kesterson dewatered sediments. Sci. Total Environ. 198:259-270. Focardi, S., C. Fossi, M. Lambertini, C. Leonzio and A. Massi. 1988. Long term monitoring of pollutants in eggs of yellow-legged herring gull from Capraia Island (Tuscan Archipelago). Environ. Monitor. Assess. 10(1):43-50. Foe, C. and A.W. Knight. Manuscript. Selenium bioaccumulation, regulation, and toxicity in the green alga, Selenastrum capricornutum , and dietary toxicity of the contaminated alga to Daphnia magna . Department of Land, Air and Water Resources, University of California, Davis, CA. Follett, R.H. 1991. Extension's Response to Reports of Toxic Levels of Selenium in Colorado Soil Plant and Water Samples. J. Agron. Ed.20(2):151-152. Foltinova, P. and J. Gajdosova. 1993. Effect of ascorbic acid and selenium on bleaching activity of furadantin and furazolidone inEuglena gracilis . Biologia (Bratislava) 48(3):291-293. Foltinova, P., N. Lahitova and L. Ebringer. 1994. Antimutagenicity in Euglena gracilis . Mutat. Res. 323(4):167-171. Forsythe, B.L. II and S.J. Klaine. 1994. The interaction of sulfate and selenate (Se+6) effects on brine shrimp, Artemia spp . Chemosphere 29(4):789-800. Fowler, B.A., R.C. Fay, R.L. Walter, R.D. Willis and W.F. Gutknecht. 1975. Levels of toxic metals in marine organisms collected from southern California coastal waters. Environ. Health Perspect. 12:71-76. Fowler, B.A., N.G. Carmichael, K.S. Squibb and D.W. Engel. 1981. Factors affecting trace metal uptake and toxicity to estuarine organisms. II. Cellular mechanisms. In: Biological monitoring of marine pollutants. Vernberg, J., A. Calabrese, F.P. Thurberg and W.B. Vernburg (Eds.). Academic Press, New York, NY. pp. 145-163. Fowler, S.W. 1986. Trace metal monitoring of pelagic organisms from the open Mediterranean Sea. Environ. Monit. Assess. 7(1):59-78. Fowler, S.W. and G. Benayoun. 1976a. Influence of environmental factors on selenium flux in two marine invertebrates. Mar. Biol. (Berl.) 37:59-68.

### Draft November 12, 2004 Ref-27 Fowler, S.W. and G. Benayoun. 1976b. Accumulation and distribution of selenium in mussels and shrimp tissue. Bull. Environ. Contam. Toxicol. 16:339-346. Fowler, S.W. and G. Benayoun. 1976c. Selenium kinetics in marine zooplankton. Mar. Sci. Commun. 2:43-67. Fowler, S.W., C. Papadopoulou and D. Zafiropoulos. 1985. Trace elements in selected species of zooplankton and nekton from the open Mediterranean Sea. Heavy Met. Environ., Int. Conf., 5th, Volume 1, 670-2. Editor(s): Lekkas, Themistokles D. CEP Consult.: Edinburgh, UK. France, R.L. 1987. Calcium and trace metal composition of crayfish ( Orconectes virilis) in relation to experimental lake acidification. Can. J. Fish. Aquat. Sci. 44(Suppl. 1):107-113. Franson, J. C., N.J. Thomas, M.R. Smith, A.H. Robbins, S. Newman, and P.C. Mccartin. 1996. A retrospective study of postmortem findings in red-tailed hawks. J. Raptor Res. 30(1):7-14. Frausto da Silva, J.J.R., and R.J. P. Williams. 1991. The Biological Chemistry of the Elements. Clarendon Press, Oxford, England. Freeman, H.C. and G.B. Sangalang. 1977. A study of the effects of methyl mercury, cadmium, arsenic, selenium, and a PCB (Aroclor 1254) on adrenal and testicular steroidogeneses in vitro, by the gray seal Halichoerus gyrpus . Arch. Environ. Contam. Toxicol. 5:369-383. Friberg, L. 1988. The GESAMP evaluation of potentially harmful substances in fish and other seafood with special reference to carcinogenic substances. Aquat. Toxicol., 11(3-4):379-93. Fries, L. 1982. Selenium stimulates growth of marine macroalgae in axenic culture. J. Phycol. 18:328-331. Froslie, A., G. Norheim and O.T. Sandlund. 1985. Levels of selenium in relation to levels of mercury in fish from Mjosa, a freshwater lake in southeastern Norway. Bull. Environ. Contam. Toxicol. 34:572-577. Froslie, A., G. Holt, R. Hoie and A Haugen. 1987. Levels of copper, selenium and zinc in liver of Norwegian moose (

Alces alces), reindeer (

Rangifer tarandus ), roe deer (Capreolus capreolus ) and hare (Lepus timidus ). Norsk Landbruksforsking 1(4): 243-250. Frost, D.V. and D. Ingvoldstad. 1975. Ecological aspects of selenium and tellurium in human and animal health. Chem. Scr. 8A:96-107. Fujita, M., M. Ike, S. Nishimoto, K. Takahashi and M. Kashiwa. 1997. Isolation and characterization of a novel selenate-reducing bacterium, Bacillussp. SF-1. J. Fermentation Bioeng. 83(6):517-522. Furr, A.K., T.F. Parkinson, W.D. Youngs, C.O. Berg, W.H. Gutenmann, I.S. Pakkala and D.J. Lisk. 1979. Elemental content of aquatic organisms inhabiting a pond comtaminated with coal fly ash. N. Y. Fish Game J. 26:154-161. Gabrashanske, M. P. and A. P. Daskalova. 1985. On the microelement composition of tissues of young geese experimentally invaded with Ascaridia galli . Helminthologia (Bratislava) 22(4):267-275.

### Draft November 12, 2004 Ref-28 Gabrashanske, M. and I. Nedeva. 1994. Microelement concentration of the host-parasite system Cyprinus carpio -cestode. Biotechnol. Equip.(4):54-57. Gaikwad, S.A. 1989. Acute toxicity of mercury, copper and selenium to the fish Etroplus maculatus

. Environ. Ecol. 7(3):694-696. Galgan, V. and A. Frank. 1995. Survey of bioavailable selenium in Sweden with the moose ( Alces alces L.) as monitoring animal. Sci. Total Environ. 172(1):37-45. Ganther, H.E. 1980. Interactions of vitamin E and selenium with mercury and silver. Ann. N. Y. Acad. Sci. 355:212-226. Gao, S. and K.K. Tanji. 1995. Model for biomethylation and volatilization of selenium from agricultural evaporation ponds. J. Environ. Qual. 24(1):191-197. Garcia-Hernandez, J., E.P. Glenn, J. Artiola, and D.J. Baumgartner. 2000. Bioaccumulation of selenium (Se) in the Cienega de Santa Clara Wetland, Sonora, Mexico. Ecotoxicol. Environ. Safety. 46:298-304. Garrett, G.P. and C.R. Inman. 1984. Selenium-induced changes in fish populations in a heated reservoir. Proc. Annu. Conf. Southeast. Assoc. Fish. Wildl. Agencies 38:291-301. Gatlin, D.M. 1983. Dietary magnesium, zinc, selenium and manganese requirements of fingerling channel catfish. Ph.D. thesis. Mississippi State University, Mississippi State, MS. Available from: University Microfilms, Ann Arbor, MI. Order No. 84-13943. Gatlin, D.M. and R.P. Wilson. 1984. Dietary selenium requirement of fingerling channel catfish. J. Nutr. 114:627-633. Gatlin, D.M. III, W.E. Poe and R.P. Wilson. 1986. Effects of singular and combined dietary deficiencies of selenium and vitamin E on fingerling channel catfish ( Ictalurus punctatus ). J. Nutr. 116(6):1061-1067. Gennity, J.M., N.R. Bottino, R.A. Zingaro, A.E. Wheeler and K.J. Irgolic. 1984. The binding of selenium to the lipids of two unicellular marine algae. Biochem. Biophys. Res. Commun. 118:176-182. Gennity, J.M., N.R. Bottino, R.A. Zingaro, A.E. Wheeler and K.J. Irgolic. 1985a. A selenium-induced peroxidation of glutathione in algae. Phytochem. 24:2817-2821. Gennity, J.M., N.R. Bottino, R.A. Zingaro. A.E. Wheeler and K.J. Irgolic. 1985b. A selenite-induced decrease in the lipid content of a red alga. Phytochem. 24:2823-2830. Gerhardt, M.B. 1990. Chemical transformations in an algal-bacterial selenium removal system. 337 pp. Avail. Univ. Microfilms Int., Order No. DA9103694 From: Diss. Abstr. Int. B 1991, 51(9):4494. Gerhardt, M.B., F.B. Green, R.D. Newman, T.J. Lundquist, R.B. Tresan and W.J. Oswald. 1991. Removal of selenium using a novel algal-bacterial process. Res. J. Water Pollut. Control Fed. 63(5):799-805.

### Draft November 12, 2004 Ref-29 Gharieb, M.M., S.C. Wilkinson, and G.M. Gadd. 1995. Reduction of Selenium Oxyanions by Unicellular Polymorphic and Filamentous Fungi: Cellular Location of Reduced Selenium and Implications for Tolerance. Journal of Industrial Microbiology . 14:300- . Giardina, B., M.L. Gozzo, B. Zappacosta, L. Colacicco, C. Calla, A. Mordente, and S. Lippa. 1997. Coenzyme Q homologs and trace elements content of Antarctic fishes Chionodraco hamatusand Pagothenia bernacchii

compared with the Mediterranean fish Mugil cephalus . Comp. Biochem. Physiol. 118A(4): 977-980. Gibbs, P.J. and A.G. Miskiewicz. 1995. Heavy metals in fish near a major primary treatment sewage plant outfall. Mar. Pollut. Bull. 30(10):667-674. Gillespie, R.B. and P. C. Baumann. 1986. Effects of high tissue concentrations of selenium on reproduction by bluegills (

Lepomis macrochirus). Trans. Am. Fish. Soc. 115(2):208-213. Gillespie, R.B., P.C. Baumann, and C.T. Singley. 1988. Dietary exposure of bluegills ( Lepomis macrochirus) to (75) Se: uptake and distribution in organs and tissues. Bull. Environ. Contam. Toxicol. 40:771-778. Gissel-Nielsen, G. and M. Gissel-Nielsen. 1973. Ecological effects of selenium application to field crops. Ambio 2:114-117. Gissel-Nielsen, G. and M. Gissel-Nielsen. 1978. Sensitivity of trout to chronic and acute exposure to selenium. Agric. Environ. 4:85-91. GLEC. 1998. Effect of sulfate concentratin on acute toxicity of selenite and selenate to invertebrates and fish. Final Report TR-111878 to the Electric Power Research Institute. GLEC. 1999. Toxicity testing and chemical analysis of selenium from acute toxicity tests. Final Report to the U.S. Environmental Protection Agency. 40pp. Glickstein, N. 1978. Acute toxicity of mercury and selenium to Crassostrea gigasembryos and Cancer magister larvae. Mar. Biol. (Berl.) 49:113-117. Gochfeld, M. 1997. Spatial patterns in a bioindicator: Heavy metal and selenium concentration in eggs of herring gulls (

Larus argentatus ) in the New York Bight. Arch. Environ. Contam. Toxicol. 33(1):63-70. Goede, A.A. 1985. Mercury, selenium, arsenic and zinc in waders from the Dutch Wadden Sea. Environ. Pollut. Ser. A Ecol. Biol. 37(4):287-310. Goede, A.A. 1991. The Variability and Significance of Selenium Concentrations in Shorebird Feathers. Environ. Monitor. Assess. 18(3):203-210. Goede, A.A. 1993a. Selenium In Eggs And Parental Blood Of a Dutch Marine Wader. Arch. Environ. Contam. Toxicol. 25(1):79-84. Goede, A.A. 1993b. Selenium status in Charadriiformes: Tissue distribution and seasonal, geographical,

### Draft November 12, 2004 Ref-30 and species variation. Biol. Trace Element Res. 39(2-3):177-190. Goede, A.A. and M. DeBruin. 1984. The use of bird feather parts as a monitor for metal pollution. Environ. Pollut. Ser. B Chem. Phys. 8(4):281-298. Goede, A.A. and M. DeBruin. 1985. Selenium in a shore bird, the dunlin ( Calidris alpina ), from the Dutch Wadden zee. Mar. Pollut. Bull. 16(3):115-117. Goede, A.A. and H.T. Wolterbeek. 1993. The bioavailability of various selenium compounds to a marine wading bird. Biol. Trace Element Res. 39(2-3):191-201. Goede, A.A. and H.T. Wolterbeek. 1994a. Have high selenium concentrations in wading birds their origin in mercury? Sci. Total Environ. 144:247-253. Goede, A.A. and H.T. Wolterbeek. 1994b. The possible role of selenium in antioxidation in marine waders: A preliminary study. Sci. Total Environ. 144:241-246. Goede, A.A., T. Nygard, M. DeBruin, and E. Steinnes. 1989. Selenium, mercury, arsenic and cadmium in the life cycle of the dunlin, Calidris alpina , a migrant wader. Sci. Total Environ. 78(0):205-218. Goede, A.A., H.T. Wolterbeek and M.J. Koese. 1993. Selenium Concentrations in the Marine Invertebrates Macoma balthica , Mytilus edulisand Nereis diversicolor. Arch. Environ. Contam. Toxicol. 25(1):85-89. Goettl, J.P., Jr., and P.H. Davies. 1976. Water pollution studies. Job Progress Report, Federal Aid Project F-33-R-11. Colorado Division of Wildlife, Fort Collins, CO. pp. 31-34. Goettl, J.P., Jr. and P.H. Davies. 1977. Water pollution studies. Job Progress Report, Federal Aid Project F-33-R-12. Colorado Division of Wildlife, Fort Collins, CO. pp. 39-42. Goettl, J.P., Jr. and P.H. Davies. 1978. Water pollution studies. Job Progress Report, Federal Aid Project F-33-R-13. Colorado Division of Wildlife, Fort Collins, CO. pp.12-13. Gotsis, O. 1982. Combined effects of selenium/mercury and selenium/copper on the cell population of the alga Dunaliella minuta . Mar. Biol. (Berl.) 71:217-222. Graham, R.V., B.G. Blaylock, F.O. Hoffman and M.L. Frank. 1992. Comparison of selenomethionine and selenite cycling in freshwater experimental ponds. Water Air Soil Pollut. 62:25-42. Gras, N., M. Thieck, L. Munoz, S. Hurtado. 1992. Seasonal and geographical variability in some trace elements of Pacific oysters ( Crassostrea gigas ) cultured in two different bays of Northern Chile. J. Radioanal. Nucl. Chem. 161(1):135-146. Great Lakes Environmental Center. 1997.Development of Site-Specific Criteria at the Albright Power Station Ash Disposal Site. Final Report. GLEC, Columbus, OH. Green, D.E. and P.H. Albers. 1997. Diagnostic criteria for selenium toxicosis in aquatic birds: Histologic lesions. J. Wild. Diseases 33(3):385-404.

### Draft November 12, 2004 Ref-31 Greenberg, A.J. and D. Kopec. 1986. Decline of Bay-Delta fisheries and increased selenium loading: Possible correlation? In: Selenium and agricultural drainage: Implications for San Francisco Bay and the California environment. The Bay Institute of San Francisco, Tiburon, CA. pp. 69-81. Greig, R.A. and J. Jones. 1976. Nondestructive neutron activation analysis of marine organisms collected from ocean dump sites of the middle eastern United States. Arch. Environ. Contam. Toxicol. 4:420-434. Grubor-Lajsic, G., A. Jovanovic, S. Maletin, M. Matavulj and A. Matic. 1995. Effects of dietary selenium on levels of plasma thyroid hormones (T3 and T4) in carp ( Cyprinus carpioL.). Naucni Skupovi - Srp. Akad. Nauka Umet. Od. Prir.-Mat. Nauka, 6(Conference on Selenium, 1993):115-118. Gunderson, C.A., J.M. Kostuk, M.H. Gibbs, G.E. Napolitano, L.F. Wicker, J.E. Richmond, and A.J. Stewart. 1997. Multispecies toxicity assessment of compost produced in bioremediation of an explosives-contaminated sediment. Environ. Toxicol. Chem. 16(12): 2529-2537. Gutenmann, W.H., W.D. Youngs and D.J. Lisk. 1976. Selenium in fly ash. Science 191:966-967. Gutenmann, W.H., C.A. Bache, J.B. McCahan and D.J. Lisk. 1988. Heavy metals and chlorinated hydrocarbons in marine fish products. Nutr. Rep. Int. 38(6):1157-61. Guthrie, R.K. and D.S. Cherry. 1976. Pollutant removal from coal-ash basin effluent. Water Resour. Bull. 12:889-902. Guthrie, R.K. and D.S. Cherry. 1979. Trophic level accumulation of heavy metals in a coal ash basin drainage system. Water Resour. Bull. 15:244-248. Gutierrez-Galindo, E.A., G. Flores Munoz, J.A. Villaescusa and A. Arreola Chimal. 1994. Spatial and Temporal Variations of Arsenic and Selenium in a Biomonitor ( Modiolus capax) from the Gulf of California, Mexico. Mar. Pollut. Bull. 28(5):330-333. Guven, K.C., S. Topcuoglu, D. Kut, N. Esen, N. Erenturk, N. Saygi, E. Cevher, B. Guvener and B. Ozturk. 1992. Metal Uptake by Black Sea Algae. Bot. Mar. 35(4):337-340. Hait, G.N. and A.K. Sinha. 1987. Biochemical changes associated with induction of resistance in rice seedlings to Helminthosporium oryzae by seed treatment with chemicals. Zeitschrift fuer Pflanzenkrankheiten und Pflanzenschutz 94(4):360-368. Halbrook, R.S., A. Woolf, G.F. Hubert Jr., S. Ross and W.E. Braselton. 1996. Contaminant concentrations in Illinois mink and otter. Ecotoxicol. 5(2):103-114. Hall, L.W, Jr. 1988. Studies of striped bass in three Chesapeake Bay spawning habitats. Mar. Pollut. Bull. 19(9):478-87. Hall, L.W., Jr., and D.T. Burton. 1982. Effects of power plant coal pile and coal waste runoff and leachate on aquatic biota: An overview with research recommendations. Crit. Rev. Toxicol. 10:287-301. Hall, L.W., Jr., L.O. Horseman and S. Zeger. 1984. Effects of organic and inroganic chemical

### Draft November 12, 2004 Ref-32 contaminants on fertilization, hatching success, and prolarval survival of striped bass. Arch. Environ. Contam. Toxicol. 13:723-729. Hall, L.W., Jr., A.E. Pinkney, R.L. Herman and S.E. Finger. 1987. Survival of striped bass larvae and yearlings in relation to contaminants and water quality in the upper Chesapeake Bay. Arch. Environ. Contam. Toxicol. 16(4):391-400. Hall, L.W., Jr, S.J. Bushong, M.C. Ziegenfuss, W.S. Hall and R.L. Herman. 1988. Concurrent mobile on-site and in situ striped bass contaminant and water quality studies in the Choptank River and Upper Chesapeake Bay. Environ. Toxicol. Chem. 7(10):815-30. Hall, L.W., Jr., M.C. Ziegenfuss and S.A. Fischer. 1992. Ambient toxicity testing in the Chesapeake Bay watershed using freshwater and estuarine water column tests. Environ. Toxicol. Chem. 11(10):1409-1425. Hall, S.L. and F.M. Fisher, Jr. 1985. Heavy metal concentrations of duck tissues in relation to ingestion of spent shot. Bull. Environ. Contam. Toxicol. 35(2):163-72 Halter, M.T., W.J. Adams and H.E. Johnson. 1980. Selenium toxicity to Daphnia magna , Hyallela azteca , and the fathead minnow in hard water. Bull. Environ. Contam. Toxicol. 24:102-107. Halverson, A.W. and K.J. Monty. 1960. An effect of dietary sulfate on selenium poisoning in the rat. J. Nutr. 70:100-102. Hamilton, S.J. 1995. Hazard assessment of inorganics to three endangered fish in the Green River, Utah. Ecotoxicol. Environ. Safety 30(2):134-142. Hamilton, S.J. (In preparation). Rationale for a tissue-based selenium criterion for aquatic life. Aquatic Toxicol. Hamilton, S.J. and K.J. Buhl. 1990a. Safety assessment of selected inorganic elements to fry of chinook salmon Oncorhynchus tshawytscha . Ecotoxicol. Environ. Safety 20(3):307-324. Hamilton, S.J. and K.J. Buhl. 1990b. Acute toxicity of boron, molybdenum and selenium to fry of chinook salmon and coho salmon. Arch. Environ. Contam. Toxicol. 19(3):366-373. Hamilton, S.J. and K.J. Buhl. 1997a. Hazard assessment of inorganics, individually and in mixtures, to two endangered fish in the San Juan, New Mexicio. Environ. Toxicol. Water Qual. 12:195-209. Hamilton, S.J. and K.J. Buhl. 1997b. Hazard evaluation of inorganics, singly and in mixtures to flannelmouth sucker Catastomos latipinnis in the San Juan, New Mexico. Ecotox. Environ. Safety 38:296- 308. Hamilton, S.J. and A.D. Lemly. 1999. Water-sediment controversy in setting environmental standards for selenium. Ecotoxicol. Environ. Safety. 44:227-235. Hamilton, S.J. and B. Waddell. 1994. Selenium in eggs and milt of razorback sucker ( Xyrauchen texanus) in the middle Green River, Utah. Arch. Environ. Contam. Toxicol. 27(2):195-201.

### Draft November 12, 2004 Ref-33 Hamilton, S.J. and R.H. Wiedmeyer. 1990. Concentrations of boron, molybdenum and selenium in chinook salmon. Trans. Am. Fisheries Soc. 119(3):500-510. Hamilton, S.J., A.N. Palmisano, G.A. Wedemeyer and W.T. Yasutake. 1986. Impacts of selenium on early life stages and smoltification of fall chinook salmon. In: Transactions of the fifty-first North American wildlife and natural resources conference. McCabe, R.E. (Ed.). Wildlife Management Institute, Washington DC. pp. 343-356. Hamilton, S.J., K.J. Buhl, N.L. Faerber, R.H. Wiedmeyer and F.A. Bullard. 1990. Toxicity of Organic Selenium in the Diet to Chinook Salmon. Environ. Toxicol. Chem. 9(3):347-358. Hamilton, S.J. K.J. Buhl, F.A. Bullard and E.E. Little. 2000. Chronic toxicity and hazard assessment of an inorganic mixture simulating irrigation drainwater to razorback sucker and bonytail. Environ. Toxicol. Chem. 15:48-64. Hamilton, S.J., K.M. Holley, K.J. Buhl., F.A. Bullard, L.K. Weston, and S.F. McDonald 2001a. The evaluation of contaminant impacts on razorback sucker held in flooded bottomland sites near Grand Junction, Colorado - 1996. Final Report. U.S. Geological Survey, Yankton, South Dakota. 302 pages. Hamilton, S.J., K.M. Holley, K.J. Buhl., F.A. Bullard, L.K. Weston, and S.F. McDonald 2001b. The evaluation of contaminant impacts on razorback sucker held in flooded bottomland sites near Grand Junction, Colorado - 1997. Final Report. U.S. Geological Survey, Yankton, South Dakota. 229 pages. Hansen, C.T., C.O. Nielsen, R. Dietz, and M.M. Hansen. 1990. Zinc, cadmium, mercury and selenium in Minke Whales, Belugas and Narwhals from West Greenland Arctic Ocean. Polar Biol. 10(7):529-540. Hansen, L.D., K.J. Maier and A.W. Knight. 1993. The effect of sulfate on the bioconcentration of selenate by Chironomus decorusand Daphnia magna . Arch. Environ. Contam. Toxicol. 25(1):72-78. Hanson, P.J. 1997. Response of hepatic trace element concentrations in fish exposed to elemental and organic contaminants. Estuaries 20(4):659-676. Hanson, P.J. and D.W. Evans. 1991. Metal contaminant assessment for the Southeast Atlantic and Gulf of Mexico coasts: results of the National Benthic Surveillance Project over the first four years 1984-87. Report, NOAA-TM-NMFS-SEFSC-284; Order No. PB92-137835, 218 pp. Avail. NTIS From: Gov. Rep. Announce. Index (U. S.) 1992, 92(8), Abstr. No. 220,226. Hardiman, S. and B. Pearson. 1995. Heavy Metals, TBT and DDT in the Sydney rock oyster ( Saccostrea commercialis) sampled from the Hawkesbury River Estuary, NSW, Australia. Mar. Pollut. Bull. 30(8):563-567. Hardy, R.W. 2002. Effects of dietary selenium on cutthroat trout ( Oncorhychus clarki) growth and reproductive performance. Report to Montgomery Watson Harza, 2375 130 th

### Ave. NE, Bellevue, WA 98005. Hargrave, B.T., P. Germain, J.C. Philippot, G. Hemon and J.N. Smith. 1992. Stable elements and polonium-210 in the deep-sea amphipod Eurythenes gryllus . Deep-Sea Res. Part A, 39(1A):37-44.

### Draft November 12, 2004 Ref-34 Harrison, P.J, P.W. Yu, P.A. Thompson, N.M. Price and D.J. Phillips. 1988. Survey of selenium requirements in marine phytoplankton. Mar. Ecol. Prog. Ser. 47(1):89-96. Harrison, S.E. and J.F. Klaverkamp. 1990. Metal contamination in liver and muscle of northern pike ( Esox lucius ) and white sucker ( Catostomus commersoni) and in sediments from lakes near the smelter at Flin Flon, Manitoba. Environ. Toxicol. Chem. 9(7):941-956. Harrison, S.E., J.F. Klaverkamp, and R.H. Hesslein. 1990. Fates of Metal Radiotracers Added to a Whole Lake Accumulation In Fathead Minnow Pimephales promelas

and Lake Trout Salvelinus namaycush

. Water Air Soil Pollut. 52(3-4):277-294. Hartwell, S.I., D.S. Cherry and J. Cairns Jr. 1987a. Avoidance responses of schooling fathead minnows (Pimephales promelas) to a blend of metals during a 9-month exposure. Environ. Toxicol. Chem. 6(3):177-188. Hartwell, S.I., D.S. Cherry and J. Cairns Jr. 1987b. Field validation of avoidance of elevated metals by fathead minnows (

Pimephales promelas) following in situ acclimation. Environ. Toxicol. Chem. 6(3):189-200. Hartwell, S.I, D. Cherry and J. Cairns Jr. 1988. Fish behavioral assessment of pollutants. ASTM Spec. Tech. Publ., 988. Funct. Test. Aquat. Biota Estim. Hazards Chem. 138-65. Hartwell, S.I., J.H. Jin, D.S. Cherry and J. Cairns, Jr. 1989. Toxicity versus avoidance response of golden shiner, Notemigonus crysoleucas , to five metals. J. Fish Biol. 35(3):447-456. Hartwell, S.I., C.E. Dawson, E.Q. Durell, R.W. Alden, P.C. Adolphson, D.A. Wright, G.M. Coelho, J.A. Magee, S. Ailstock, and M. Novman. 1997. Correlation of measures of ambient toxicity and fish community diversity in Chesapeake Bay, USA, tributaries - urbanizing watersheds. Environ. Toxicol. Chem. 16(12):2556-2567. Hasunuma, R., T. Ogawa, Y. Fujise and Y. Kawanishi. 1993. Analysis of selenium metabolites in urine samples of Minke WhaleBalaenoptera acutorostrata using ion exchange chromatography. Comp. Biochem. Physiol. - C:Comparative Pharmacol. & Toxicol. 104(1):87-89. Hatcher, C.O., R.E. Ogawa, T.P. Poe and J.R.P. French III. 1992. Trace elements in lake sediment macrozoobenthos and fish near a coal ash disposal basin. J. Freshwater Ecol. 7(3):257-269. Haynes, D., J. Leeder and P. Rayment. 1995. Temporal and spatial variation in heavy metal concentrations in the bivalve Donax deltoidesfrom the Ninety Mile Beach, Victoria, Australia. Mar. Pollut. Bull. 30(6):419-424. Hayward, D.G., M.X. Petreas, J.J. Winkler, P. Visita, M. McKinney, R.D. Stephens. 1996. Investigation of a wood treatment facility: impact on an aquatic ecosystem in the San Joaquin River, Stockton, California. Arch. Environ. Contam. Toxicol. 30(1):30-39. Heider, J., and A. Boeck. 1993. Selenium Metabolism in Micro-Organisms. Advances in Microbial Physiology . 35:71- .

### Draft November 12, 2004 Ref-35 Hein, R.G., P.A. Talcott, J.L. Smith and W.L. Myers. 1994. Blood selenium values of selected wildlife populations in Washington. Northwest Science 68(3):185-188. Heiny, J.S. and C.M. Tate. 1997. Concentration, distribution, and comparison of selected trace elements in bed sediment and fish tissue in the South Platte River Basin, USA, 1992-1993. Arch. Environ. Contam. Toxicol. 32(3):246-259. Heinz, G.H. 1993a. Selenium accumulation and loss in mallard eggs. Environ. Toxicol. Chem. 12(4):775-778. Heinz, G.H. 1993b. Re-exposure of mallards to selenium after chronic exposure. Environ. Toxicol. Chem. 12(9):1691-1694. Heinz, G.H. and M.A. Fitzgerald. 1993a. Reproduction of mallards following overwinter exposure to selenium. Environ. Pollut. 81(2):117-122. Heinz, G.H. and M.A. Fitzgerald. 1993b. Overwinter survival of mallards fed selenium. Arch. Environ. Contam. Toxicol. 25(1):90-94. Heinz, G.H. and L.G. Gold. 1987. Behavior of mallard ducklings from adults exposed to selenium. Environ. Toxicol. Chem. 6(11):863-866. Heinz, G.H. and D.J. Hoffman. 1996. Comparison of the effects of seleno-L-methionine, seleno-DL-methionine, and selenized yeast on reproduction of mallards. Environ. Pollut. 91(2):169-175. Hienz, G.H. and D.J. Hoffman. 1998. Methylmercury chloride and selenomethionine interaction on health and reproduction in mallards. Environ. Toxicol. Chem. 17(2): 139-145. Heinz, G.H. and C.J. Sanderson. 1990. Avoidance of selenium-treated food by mallards. Environ. Toxicol. Chem. 9(9):1155-1158. Heinz, G.H., D.J. Hoffman, A.J. Krynitsky and D.M.G. Weller. 1987. Reproduction in mallards fed selenium. Environ. Toxicol. Chem. 6:423-433. Heinz, G.H., D.J. Hoffman, and L.G Gold. 1988. Toxicity of organic and inorganic selenium to mallard ducklings. Arch. Environ. Contam. Toxicol. 17(5):561-8. Heinz, G.H., D.J. Hoffman, and L.G. Gold. 1989. Impaired reproduction of mallards fed an organic form of selenium. J. Wildlife Manag. 53(2):418-428. Heinz, G.H., G.W. Pendleton, A.J. Krynitsky and L.G. Gold. 1990. Selenium accumulation and elimination in mallards. Arch. Environ. Contam. Toxicol. 19(3):374-379. Heinz, G. H., D.J. Hoffman, and L.J. Lecaptain. 1996. Toxicity of seleno-L-methionine, seleno-DL-methionine, high selenium wheat, and selenized yeast to mallard ducklings. Arch. Environ. Contam. Toxicol. 30(1):93-99.

### Draft November 12, 2004 Ref-36 Heisinger, J.F. 1981. Antagonism of selenium and cadmium pretreatments to subsequent embryotoxic doses of mercury and cadmium in fish embryos. PB82-256645. National Technical Information Service, Springfield, VA. Heisinger, J.F. and S.M. Dawson. 1983. Effect of selenium deficiency on liver and blood glutathione peroxidase activity in the black bullhead. J. Exp. Zool. 225:325-327. Heisinger, J.F. and L. Scott. 1985. Selenium prevents mercuric chloride-induced acute osmoregulatory failure without glutathione peroxidase involvement in the black bullhead ( Ictalurus melas). Comp. Biochem. Physiol. C Comparative. Toxicol. 80(2):295-297. Heisinger, J.F. and E. Wait. 1989. The effects of mercuric chloride and sodium selenite on glutathione and total nonprotein sulfhydryls in the kidney of the black bullhead Ictalurus melas . Comp. Biochem. Physiol. C: Comparative Pharmacol. Toxicol. 94(1):139-142. Heisinger, J.F., C.D. Hansen and J.H. Kim. 1979. Effect of selenium dioxide on the accumulation and acute toxicty of mercuric chloride in goldfish. Arch. Environ. Contam. Toxicol. 8:279-283. Heit, M. 1985. Concentrations of potentially toxic trace elements in the muscle tissue of fish from acidic and circumneutral Adirondack lakes. Heavy Met. Environ., Int. Conf., 5th, Volume 1, 655-7. Editor(s): Lekkas, Themistokles D. CEP Consult.: Edinburgh, UK. Heit, M. and C.S. Klusek. 1985. Trace element concentrations in the dorsal muscle of white suckers and brown bullheads from two acidic Adirondack lakes. Water Air Soil Pollut. 25:87-96. Heit, M., C.S. Klusek and K.M. Miller. 1980. Trace element, radionuclide, and polynuclear aromatic hydrocarbon concentrations in Unionidae mussels from northern Lake George. Environ. Sci. Technol. 14:465-468. Heit, M., C. Schofield, C.T. Driscoll, and S.S. Hodgkiss. 1989. Trace element concentrations in fish from three Adirondack Lakes (New York, USA) with different pH values. Water Air Soil Pollut. 44(1-2):9-30. Heitmuller, P.T., T.A. Hollister and P.R. Parrish. 1981. Acute toxicity of 54 industrial chemicals to sheepshead minnows (

Cyprinodon variegatus ). Bull. Environ. Contam. Toxicol. 27:596-604. Hellou, J., W.G. Warren, J.F. Payne, S. Belkhode and P. Lobel. 1992a. Heavy Metals and Other Elements in Three Tissues of Cod Gadus morhuafrom the Northwest Atlantic. Mar. Pollut. Bullet. 24(9):452-458. Hellou, J., L.L. Fancey and J.F. Payne. 1992b. Concentrations of twenty-four elements in bluefin tuna, Thunnus thynnusfrom the Northwest Atlantic. Chemosphere 24(2):211-218. Hellou, J., V. Zitko, J. Friel and T. Alkanani. 1996a. Distribution of elements in tissues of yellowtail flounder Pleuronectes ferruginea. Sci. Total Environ. 181(2):137-146. Hellou, J., J. Banoub, C. Andrews, D. Gentil, V. O'Malley, T. Biger, C. Barno, and D. House. 1996b. Crankcase oil, hydrocarbons, the environment and rainbow trout. Can. Tech. Rep. Fish. Aquat. Sci., 2093, Proceedings of the 22nd Annual Aquatic Toxicity Workshop, 1995, 47-52.

### Draft November 12, 2004 Ref-37 Henderson, G.B., A.H. Fairlamb and A. Cerami. 1987. Trypanothione dependent peroxide metabolism in Crithidia fasciculata and Trypanosoma brucei. Molec. Biochem. Parasitol. 24(1):39-46. Henebry, M.S. and P.E. Ross. 1989. Use of protozoan communities to assess the ecotoxicological hazard of contaminated sediments. Toxic. Assess. 4(2):209-27. Henny, C.J. and J.K. Bennett. 1990. comparison of breaking strength and shell thickness as evaluators of white-faced ibis eggshell quality. Environ. Toxicol. Chem. 9(6):797-806. Henny, C.J. and G.B. Herron. 1989. DDE, selenium, mercury and White-faced Ibis Reproduction at Carson Lake Nevada USA. J. Wildlife Manag. 53(4):1032-1045. Henny, C.J., L.J. Blus, S.P. Thompson and U.W. Wilson. 1989. Environmental Contaminants Human Disturbance and Nesting of Double-crested Cormorants in Northwestern Washington USA. Colonial Waterbirds 12(2):198-206. Henny, C.J., L.J. Blus and R.A. Grove. 1990. Western GrebeAechmophorus occidentalis Wintering Biology and Contaminant Accumulation in Commencement Bay Puget Sound Washington USA. Can. Field-Naturalist 104(3):460-472. Henny, C.J., D.D. Rudis, T.J. Roffe, and E. Robinson Wilson. 1995. Contaminants and Sea Ducks in Alaska and the Circumpolar Region. Environ. Health Perspectives 103(SUPPL. 4):41-49. Hermanutz, R.O. 1992. Malformation of the fathead minnow ( Pimephales promelas) in an ecosystem with elevated selenium concentrations. Bull. Environ. Contam. Toxicol. 49(2):290-294. Hermanutz, R.O., K.N. Allen, T.H. Roush and S.F. Hedtke. 1992. Effects of elevated selenium concentrations on bluegills Lepomis macrochirusin outdoor experimental streams. Environ. Toxicol. Chem. 11(2):217-224. Hermanutz, R.O., K.N. Allen, N.E. Detenbeck, and C.E. Stephan. 1996. Exposure to bluegill ( Lepomis macrochirus) to selenium in outdoor experimental streams. U.S. EPA Report. Mid-Continent Ecology Division. Duluth, MN.
Hicks, B.D., J.W. Hilton and H.E. Ferguson. 1984. Influence of dietary selenium on the occurrence of nephrocalcinosis in the rainbow trout, Salmo gairdneriRichardson. J. Fish Dis. 7:379-389. Hildebrand, S.G., R.M. Cushman and J.A. Carter. 1976. The potential toxicity and bioaccumulation in aquatic systems of trace elements present in aqueous coal conversion effluents. In: Trace substances in environmental health - X. Hemphill, D.D. (Ed.). University of Missouri, Columbia, MO. pp. 305-312. 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. Hilton, J.W., P.V. Hodson and S.J. Slinger. 1980. The requirement and toxicity of selenium in rainbow trout (Salmo gairdneri ). J. Nutr. 110:2527-2535.

### Draft November 12, 2004 Ref-38 Hilton, J.W., P.V. Hodson and S.J. Slinger. 1982. Absorption, distribution, half-life and possible routes of elimination of dietary selenium in juvenile rainbow trout ( Salmo gairdneri ). Comp. Biochem. Physiol. 71C:49-55. Hiraika, Y., S. Ishizawa and T. Kamada. 1985. Acute toxicity of 14 different kinds of metals affecting medaka ( Oryzias latipes ) fry. Hiroshima J. Med. Sci. 34(3):327-330. Hjeltnes, B. and K. Julshman. 1992. Concentrations of iron, copper, zinc and selenium in liver of atlantic salmon Salmo salar

infected with Vibrio salmonicida . Dis. of Aquat. Organ. 12(2):147-149. Hockett, J.R. and D.R. Mount. 1996. Use of metal chelating agents to differentiate among sources of acute aquatic toxicity. Environ. Toxicol. Chem. 15(10):1687-1693. Hodge, V., K. Stetzenbach and K. Johannesson. 1996. Initial results for the inductively coupled plasma-mass spectrometric determination of trace elements in organs of striped bass from Lake Mead, U.S.A.. ACS Symp. Ser., 643:180-190. Hodson, P.V. 1990. Indicators of ecosystem health at the species level and the example of selenium effects on fish. Environ. Monit. Assess. 15(3):241-254. Hodson, P.V. and J.W. Hilton. 1983. The nutritional requirements and toxicity to fish of dietary and waterborne selenium. Ecol. Bull. 35:335-340. Hodson, P.V., D.J. Spry and B.R. Blunt. 1980. Effects on rainbow trout ( Salmo gairdneri ) of a chronic exposure to waterborne selenium. Can. J. Fish. Aquat. Sci. 37:233-240. Hodson, P.V., D.M. Whittle and D.J. Hallett. 1984. Selenium contamination of the Great Lakes and its potential effects on aquatic biota. In: Toxic contaminants in the Great Lakes. Nriagu, J.O. and M.S. Simmons (Eds.). Wiley, New York, NY. pp. 371-391. Hodson, P.V., J.W. Hilton and S.J. Slinger. 1986. Accumulation of waterborne selenium by rainbow trout (Salmo gairdneri ), eggs, fry and juveniles. Fish Physiol. Biochem. 1(4):187-96. Hoffman, D. and G.H. Heinz. 1987. Developmental toxicity of excess selenium in mallard ( Anas platyrhynchos) ducks. Fed. Proc. 46:1154. Hoffman, D.J. and G.H. Heinz. 1988. Embryotoxic and teratogenic effects of selenium in the diet of mallards. J. Toxicol. Environ. Health 24(4):477-490. Hoffman, D. and G.H. Heinz. 1998. Effects of mercury and selenium on glutathione metabolisms and oxidative stress in mallard ducks. Environ. Toxicol. Chem. 17(2):161-166. Hoffman, D.J., Ohlendorf, H.M. and T.W. Aldrich. 1988. Selenium teratogenesis in natural populations of aquatic birds in central California (USA). Arch. Environ. Contam. Toxicol. 17(4):519-526. Hoffman, D.J., G.H. Heinz, and A.J. Krynitsky. 1989. Hepatic glutathione metabolism and lipid peroxidation in response to excess dietary selenomethionine and selenite in mallard ducklings. J. Toxicol.

### Draft November 12, 2004 Ref-39 Environ. Health 27(2):263-272. Hoffman, D.J., C.J. Sanderson, L.J. Lecaptain, E. Cromartie and G.W. Pendelton. 1991a. Interactive Effects of Boron, Selenium and Dietary Protein on Survival Growth and Physiology in Mallard Ducklings. Arch. Environ. Contam. Toxicol. 20(2):288-294. Hoffman, D.J., G.H. Heinz, L.J. LeCaptain, C.M. Bunck and D.E. Green. 1991b. Subchronic hepatotoxicity of selenomethionine ingestion in mallard ducks. J. Toxicol. Environ. Health 32(4):449-464. Hoffman, D.J., C.J. Sanderson, L.J. Lecaptain, E. Cromartie and G.W. Pendleton. 1992a. Interactive Effects Of Selenium Methionine And Dietary Protein On Survival Growth And Physiology In Mallard Ducklings. Arch. Environ. Contam. Toxicol. 23(2):162-171. Hoffman, D. J., C.J. Sanderson, L.J. Lecaptain, E. Cromartie and G.W. Pendleton. 1992b. Interactive Effects Of Arsenate Selenium And Dietary Protein On Survival Growth And Physiology In Mallard Ducklings. Arch. Environ. Contam. Toxicol. 22(1):55-62. Hoffman, D.J., G.H. Heinz, L.J. Lecaptain, J.D. Eisemann and G.W. Pendleton. 1996. Toxicity and oxidative stress of different forms of organic selenium and dietary protein in mallard ducklings. Arch. Environ. Contam. Toxicol. 31(1):120-127. Hoffman, D.J., H.M. Ohlendorf, C.M. Marn and G.W. Pendelton. 1998. Association of mercury and selenium with altered glutathione metabolism and oxidative stress in diving ducks from San Francisco Bay region, USA. Environ. Toxicol. Chem. 17(2):167-172. Hoglund, J. 1991. Ultrastructural observations and radiometric assay on cercarial penetration and migration of the digenean Diplostomum spathaceumin the rainbow trout Oncorhynchus mykiss . Parasitol. Res. 77(4):283-289. Hollibaugh, J.T., D.L.R. Seibert and W.H.Thomas. 1980. A comparison of the acute toxicities of ten heavy metals to phytoplankton from Saanich Inlet, B.C., Canada. Estuarine Coastal Mar. Sci. 10(1):93-105. 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 26 th

### Annual Larval Fish Conference 2003, Bergen, Norway. ISBN 82-7461-059-B. Homziak, J., L. Bennett, P. Simm and R. Herring. 1993. Metal leaching from experimental coal fly-ash oyster cultch. Bull. Environ. Contam. Toxicol. 51(2):317-324. Honda, K., Y. Fujise, R. Tatsukawa, K. Itano and N. Miyazaki. 1986. Age-related accumulation of heavy metals in bone of the striped dolphin, Stenella coeruleoalba . Mar. Environ. Res. 20(3):143-60. Hontela, A., P. Dumont, D. Duclos and R. Fortin. 1995. Endocrine and metabolic dysfunction in yellow

### Draft November 12, 2004 Ref-40 perch, Perca flavescens, exposed to organic contaminants and heavy metals in the St. Lawrence river. Environ. Toxicol. Chem. 14(4):725-731. Horne, A.J. 1991. Selenium detoxification in wetlands by permanent flooding: I. Effects on a macroalga, an epiphytic herbivore, and an invertebrate predator in the long-term mesocosm experiment at Kesterson Reservoir, California. Water Air Soil Pollut. 57-58:43-52. Hothem, R.L. and H.M. Ohlendorf. 1989. Contaminants in Foods of Aquatic Birds at Kesterson Reservoir California USA 1985. Arch. Environ. Contam. Toxicol. 18(6):773-786. Hothem, R.L. and D. Welsh. 1994a. Duck and shorebird reproduction in the grasslands of central California. Calif. Fish Game 80(2):68-79. Hothem, R.L. and D. Welsh. 1994b. Contaminants in eggs of aquatic birds from the grasslands of Central California. Arch. Environ. Contam. Toxicol. 27(2):180-185. Hothem, R.L. and S.G. Zador. 1995. Environmental contaminants in eggs of California least terns ( Sterna antillarum ). Bull. Environ. Contam. Toxicol. 55(5):658-665. Hothem, R.L., D.L. Roster, K.A. King, T.K. Keldsen, K.C. Marois, and S.E. Wainwright. 1995. Spatial and Temporal Trends Of Contaminants In Eggs Of Wading Birds From San Francisco Bay, California. Environ. Toxicol. Chem. 14(8):1319-1331. Houpt, K.A., L.A. Essick, E.B. Shaw, D.K. Alo, J.E. Gilmartin, W.H. Gutenmann, C.B. Littman, and D.J. Lisk. 1988. A tuna fish diet influences cat behavior. J. Toxicol. Environ. Health 24(2):161-172. Hsu, H.H. 1986. The uptake and distribution of radioselenium in the larvae of Fasciola hepatica

and its snail host Lymnaea columella . Vet. Parasitol. 21(4):233-45. Hsu, S.Y. and F.W. Goetz. 1992. Oxoanions stimulate in vitro ovulation and signal transduction pathways in goldfish (

Carassius auratus ) follicles. Am. J. Physiol. 263(5, Pt. 1):E943-E949. Hsu, Y. L., Y.H. Yang, Y.C. Chen, M.C. Tung, J.L. Wu, M.H. Engleking and J.C. Leong. 1995. Development of an in vitro subculture system for the oka organ (lymphoid tissue) of Penaeus monodon. Aquaculture 136(1-2):43-55. Hu, M. Y. Yang, J.M. Martin, Ko Yin, and P.J. Harrison. 1996. Preferential uptake of Se (IV) over Se (VI) and the production of dissolved organic Se by marine phytoplankton. Mar. Environ. Res. 42(2): 225- 231. Huckabee, J.W. and N.A. Griffith. 1974. Toxicity of mercury and selenium to the eggs of carp ( Cyprinus carpio ). Trans. Am. Fish. Soc. 103:822-825. Huerkamp, M.J., D.H. Ringler, and C.E. Chrisp. 1988. Vitamin E deficiency in goldfish fed a shellfish derived diet. Lab. Animal Sci. 38(2):178-182. Hunn, J.B., S.J. Hamilton and D.R. Buckler. 1987. Toxicity of sodium selenite to rainbow trout fry. Water

### Draft November 12, 2004 Ref-41 Res. 21:233-238. Hunter, C.L., M.D. Stephenson, R.S. Tjeerdema, D.G. Crosby, G.S. Ichikawa, J.D. Goetzl, K.S. Paulson, D.B. Crane, M. Martin and J.W. Newman. 1995. Contaminants in oysters in Kaneohe Bay, Hawaii. Mar. Pollut. Bull. 30(10):646-654. Hunter, D. B., P.M. Bertsch, K.M. Kemner, and S.B. Clark. 1997. Distribution and chemical speciation of metals and metalloids in biota collected from contaminated environments by spatially resolved XRF, XANES, and EXAFS. J. Phys. IV, 7(2):767-771. Hurvich, C. M., and C. Tsai. 1989. Regression and time series model selection in small samples. Biometrika . 76:297-307. Hurvich, C.M. and C. Tsai. 1990. Model selection for least absolute deviations regression in small samples. Stat. Prob. Lett. 9:259-265. Hutchinson, T.C. 1973. Comparative studies of the toxicity of heavy metals to phytoplankton and their synergistic interactions. Water Pollut. Res. Can. 8:68-90. Hutchinson, T.C. and P.M. Stokes. 1975. Heavy metal toxicity and algal bioassays. In: Water quality parameters. Barabas, S. (Ed.). ASTM STP 573. American Society for Testing and Materials, Philadelphia, PA. pp. 320-343. Hyne, R.V., A. Padovan, D.L. Parry and S.M. Renaud. 1993. Increased fecundity of the cladoceran MoinoDaphnia macleayi

on a diet supplemented with a green alga, and its use in uranium toxicity tests. Aust. J. Mar. Freshwater Res. 44(3):389-399. Ibrahim, A.M. and A. Spacie. 1990. Toxicity of inorganic selenium to the green alga Selenastrum capricornutumPrintz. Environ. Exp. Bot. 30(3):265-269. Ibrahim, H. and E. Farrag. 1992. Determination of Selenium in Biomphalaria alexandrinaSnails by Direct Current Plasma-atomic Emission Spectrometry. Microchem. J. 45(3):356-360 Ibrahim, N. and I. Mat. 1995. Trace element content in relation to the body weight of the marine bivalve, Anadara granosawith special reference to the application of INAA and ICP-AES as analytical techniques. J. Radioanal. Nucl. Chem. 195(1):203-208. Ihaka, R., and R. Gentleman. 1996. R: A Language for Data Analysis and Graphics. J. Comput. Graph. Stat 5:299-314. Ingersoll, C.G., F.J. Dwyer and T.W. May. 1990. Toxicity of inorganic and organic selenium to Daphnia magnacladocera and Chironomus riparius diptera. Environ. Toxicol. Chem. 9(9):1171-1182. Insightful Corporation. 2001. S-Plus Professional. Release 2. Seattle, Washington. Ishikawa, M., T. Ishii, S. Uchida and K. Kitao. 1987. A proton microprobe scanning across the vertebra of a flatfish, Paralichthys olivaceus. Biol. Trace Elem. Res. 13:143-57.

### Draft November 12, 2004 Ref-42 Ishikawa, M., K. Nakamura, T. Ishii, A. Bassari, K. Okoshi and K. Kitao. 1993. Elements in tissues and organs of an Antarctic fish, Champsocephalus gunnari . Nucl. Instrum. Methods Phys. Res. Sect. B, B75(1-4):204-208. Itano, K., S. Kawai, N. Miyazaki, R. Tatsukawa and T. Fujiyama. 1984. Mercury and selenium levels at the fetal and suckling stages of striped dolphin, Stenella coeruleoalba . Agric. Biol. Chem. 48(7):1691-1698. Itano, K., S. Kawai and R. Tatsukawa. 1985a. Distribution of mercury and selenium in muscle of striped dolphins. Agric. Biol. Chem. 49(2):515-17. Itano, K., S. Kawai and R. Tatsukawa. 1985b. Properties of mercury and selenium in salt-insoluble fraction of muscles in striped dolphin ( Stenella coeruleoalba). Bull. Jap. Soc. Sci. Fish. 51(7):1129-1132. Jackson, M B. 1988. The dominant attached filamentous algae of Georgian Bay, the North Channel and Eastern Lake Huron: field ecology and biomonitoring potential during 1980. Hydrobiologia 163:149-71. Jackson, M.B., E.M. Vandermeer, and L.S. Heintsch. 1990. Attached filamentous algae of northern Lake Superior: field ecology and biomonitoring potential during 1983. J. Great Lakes Res. 16(1):158-168. Jacquez, R.B., P.R. Turner, H. El-Reyes, and C.M. Lou. 1987. Characterization and treatment of wastewater generated from saline aquaculture of channel catfish. Proc. Ind. Waste Conf., Volume Date 1986, 41st, 530-8. Jakubczak, E., C. Delmaere and H. Leclerc. 1981. Sensitivity of bacteria in an aquatic environment to some toxic substances. INSERM (Inst. Nat. Santa Rech. Med.) Colloq. 106:93-104. James, G.D., S.D. Mills and G. Pattenden. 1993. Total synthesis of pukeleimide A, a 5-ylidenepyrrol-2(5H)-one from blue green algae. J. Chem. Soc., Perkin Trans. 1(21):2581-2584. Jarman, W.M., K.A. Hobson, W.J. Sydeman, C.E. Bacon and E.B. Mclaren. 1996. Influence of trophic position and feeding location on contaminant levels in the Gulf of the Farallones food web revealed by stable isotope analysis. Environ. Sci. Technol. 30(2):654-660. Jay, F.B. and R.J. Muncy. 1979. Toxicity to channel catfish of wastewater from an Iowa coal beneficiation plant. Iowa State J. Res. 54:45-50. Jayasekera, R. 1994. Pattern of distribution of selected trace elements in the marine brown alga, Sargassum filipendulaAg. from Sri Lanka. Environ. Geochem. Health 16(2):70-75. Jayasekera, R. and M. Rossbach. 1996. Use of seaweeds for monitoring trace elements in coastal waters. Environ. Geochem. Health 18(2):63-68. Jenkins, D.W. 1980. Biological monitoring of toxic trace metals. Vol. 2. Toxic trace metals in plants and animals of the world. Part III. EPA-600/3-80-092 or PB81-103509. National Technical Information Service, Springfield, VA.

### Draft November 12, 2004 Ref-43 Jenner, H. A. and T. Bowmer. 1990. The accumulation of metals and their toxicity in the marine intertidal invertebrates Cerastoderma edule, Macoma balthica and Arenicola marinaexposed to pulverized fuel ash in mesocosms. Environ. Pollut. 66(2):139-156. Jenner, H.A. and T. Bowmer. 1992. The accumulation of metals and toxic effects in Nereis virensexposed to pulverized fuel ash. Environ. Monit. Assess. 21(2):85-98. Jenner, H.A. and J.P.M. Janssen-Mommen. 1989. Phytomonitoring of pulverized fuel ash leachates by the duckweed Lemna minor . Hydrobiologia 188-189:361-366. Jenner, H.A. and J.P.M. Janssen-Mommen. 1993. Duckweed Lemna minor

as a tool for testing toxicity of coal residues and polluted sediments. Arch. Environ. Contam. Toxicol. 25(1):3-11. Jin, L.J., P. Guo, and X.Q. Xu. 1997. Effect of selenium on mercury methylation in anaerobic lake sediments. Bull. Environ. Contam. Toxicol. 59(6):994-999. Johns, C., S.N. Luoma, and V. Elrod. 1988. Selenium accumulation in benthic bivalves and fine sediments of San Francisco Bay, the Sacramento-San Joaquin Delta (USA), and selected tributaries. Estuarine Coastal Shelf Sci. 27(4):381-396. Johnson, M.G. 1987. Trace element loadings to sediments of fourteen Ontario lakes and correlations with concentrations in fish. Can. J. Fish. Aquat. Sci. 44:3-13. Johnston, P.A. 1987. Acute toxicity of inorganic selenium to Daphnia magna(Straus) and the effect of sub-acute exposure upon growth and reproduction. Aquat. Toxicol. (Amsterdam) 17(3):335-352. Johnston, P.A. 1989. Morphological changes in Daphnia magna(Straus) exposed to inorganic selenium as sodium selenate. Aquat. Toxicol. (Amsterdam) 14(2):95-108. Jones, J.B. and T.C. Stadtman. 1977. Methanococcus vanielii : Culture and effects of selenium and tungsten on growth. J. Bacteriol. 1977:1404-1406. Jonnalagadda, S.B. and P.V.V.P. Rao. 1993. Toxicity, bioavailability and metal speciation. Comp. Biochem. Physiol. 106C(3):585-595. Jop, K. M., R.C. Biever, J.R. Hoberg and S.P. Shepherd. 1997. Analysis of metals in blue crabs, Callinectes sapidus , from two Connecticut estuaries. Bull. Environ. Contam. Toxicol. 58(2):311-317. Jorgensen, D. and J. F. Heisinger. 1987. The effects of selenium on the distribution of mercury in the organs of the black bullhead ( Ictalurus melas). Comp. Biochem. Physiol. C Comp. Pharmacol. Toxicol. 87(1):181-186. Jorhem, L., J. Engman, B. Sundstrom and A.M. Thim. 1994. Trace elements in crayfish: Regional differences and changes induced by cooking. Arch. Environ. Contam. Toxicol. 26(2):137-142. Jovanovic, A., G. Grubor-Lajsic, N. Djukic, M. Telesmanic, G. Gardinovacki and M.B. Spasic. 1995. Effect of selenium supplementation on GSH-Px activity in tissues of carp ( Cyprinus carpioL.). Naucni

### Draft November 12, 2004 Ref-44 Skupovi - Srp. Akad. Nauka Umet., Od. Prir.-Mat. Nauka, 6(Conference on Selenium, 1993):99-103. Jovanovic, A., G. Grubor-Lajsic, N. Djukic, G. Gardinovacki, A. Matic and M. Spasic. 1997. The effect of selenium on antioxidant system in erythocytes and liver of the carp ( Cyprinus carpoL.). Critical Rev. Food Sci. Nutr. 37(5):443-448. Juhnke, I. and D. Ludemann. 1978. Results of the investigation of 200 chemical compounds for acute toxicity with the golden orfe test. Z. Wasser Abwasser Forsch. 11:161-164. Julshamn, K., A. Andersen, O. Ringdal and J. Morkore. 1987. Trace elements intake in the Faroe Islands (Denmark): I. Element levels in edible parts of pilot whales ( Globicephalus meleanus). Sci. Total Environ. 65(0):53-62. Julshamn, K., K. Sandnes, O. Lie and R. Waagboe. 1990. Effects of dietary selenium supplementation on growth, blood chemistry and trace element levels in serum and liver of adult Atlantic salmon ( Salmo salar). Fiskeridir. Skr., Ser. Ernaer. 3(2):47-58. Kai, N., T. Ueda, M. Takeda and A. Kataoka. 1986a. The levels of mercury and selenium in gonad of marlins from the Pacific Ocean. Bull. Jap. Soc. Scient. Fish. 52(3):553-556. Kai, N., T. Ueda, M. Takeda, Y. Takeda and A. Kataoka. 1986b. The levels of mercury and selenium in gonad of yellow fin (

Thunnus albacares ) and albacore (Thunnus alalunga ). Bull. Jap. Soc. Scient. Fish. 52(6):1049-1054. Kai, N., T. Ueda, Y. Takeda and A. Kataoka. 1988. The levels of mercury and selenium in blood of tunas. Nippon Suisan Gakkaishi 54(11):1981-5. Kai, N., T. Ueda, Y Takeda and A. Kataoka. 1992a. The levels of mercury and selenium in gonad of Big-Eyed Tuna. J. of Shimonoseki Univer. Fish. 40(4):177-181. Kai, N., T. Ueda and Y. Takeda. 1992b. The state of oxidation and its distribution of selenium in the blood of tuna and marlins. Nippon Suisan Gakkaishi 58(10):1883-1886. Kai, N., T. Tsuda, T. Sakai, H. Murata, M. Hamada, Y. Tanoue, and T. Nagai. 1995. Glutathione peroxidase activity in the blood of tunas and marlins. Fish. Sci. 61(5):867-870. Kai, N., T. Tsuda, T. Sakai, H. Murata, M. Hamada, Y. Tanoue and T. Nagai. 1996. The oxidation state and its distribution of selenium in the blood of cultured yellow tail Seriola quinqueradiata . Fish. Sci. 62(3):444-446. Kaiser, I.I., P.A. Young and J.D. Johnson. 1979. Chronic exposure of trout to waters with naturally high selenium levels: Effects on transfer RNA methylation. J. Fish. Res. Board. Can. 36:689-694. Kaiser, K.L.E. 1980. Correlation and predicition of metal toxicity to aquatic biota. Can. J. Fish. Aquat. Sci. 37:211-218. Kaiser, K.L.E., S.P. Niculescu and G. Schuurmann. 1997. Feed forward backpropagation neural networks

### Draft November 12, 2004 Ref-45 and their use in predicting the acute toxicity of chemicals to the fathead minnow. Water Qual. Res. J. Can. 32(3):637-657. Kalas, J.A., T.H. Ringsby, and S. Lierhagen. 1995. Metals and selenium in wild animals from Norwegian areas close to Russian nickel smelters. Environ. Monitor. Assess. 36(3):251-270. Kapu, M.M. and D.J. Schaeffer. 1991. Planarians in toxicology. Responses of asexualDugesia dorotocephalato selected metals. Bull. Environ. Contam. Toxicol. 47(2):302-307. Karlson, U. and W.T. Frankenberger Jr. 1990. Volatilization of selenium from agricultural evaporation pond sediments. Sci. Total Environ. 92:41-54. Kay, S.H. 1984. Potential for biomagnification of contaminants within marine and freshwater food webs. ADA150747. National Technical Information Service, Springfield, VA. Keating, K.I. and P.B. Caffrey. 1989. Selenium deficiency induced by zinc deprivation in a crustacean. Proceedings Of The National Academy Of Sciences Of The United States Of America 86(16):6436-6440. Keating, K.I. and B.C. Dagbusan. 1984. Effect of selenium deficiency on cuticle integrity in the Cladocera (Crustacea). Proc. Natl. Acad. Sci. U. S. A. 81:3433-3437. Kedziroski, A., M. Nakonieczny, E. Swierczek and E. Szulinska. 1996. Cadmium-selenium antagonism and detoxifying enzymes in insects. Fresenius J. Anal. Chem. 354(5-6):571-575. Keller, M.D., R.C. Selvin, W. Claus, and R.R.L. Guillard. 1987. Media for the culture of oceanic ultraphytoplankton. J. Phycol. 23(4):633-638. Kelly, R.K., J.F. Klaverkamp, R.V. Hunt, and O. Nielsen. 1987. Chemical analysis of muscle from walleye (Stizostedion vitreum ) with myofibrogranuloma, a chronic myopathy. Can. J. Fish. Aquat. Sci. 44(8):1425-1431. Kemble, N.E., W.G. Brumbaugh, E.L. Brunson, F.J. Dwyer, C.G. Ingersoll, D.P. Monda and D.F. Woodward. 1994. Toxicity of metal-contaminated sediments from the Upper Clark Fork River, Montana, to aquatic invertebrates and fish in laboratory exposures. Environ. Toxicol. Chem. 13(12):1985-1997. Kennedy, P.C. 1986. The use of mollusks for monitoring trace elements in the marine environment in New Zealand 1. The contribution of ingested sediment to the trace element concentrations in New Zealand mollusks. N. Z. J. Mar. Freshwater Res. 20(4):627-40. 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. Kersten, M., M. Kriews and U. Foerstner. 1991. Partitioning of trace metals released from polluted marine aerosols in coastal seawater. Mar. Chem. 36(1-4):165-182. Khan, A.H., M. Ali, S.K. Biaswas and D.A. Hadi. 1987. Trace elements in marine fish from the Bay of

### Draft November 12, 2004 Ref-46 Bengal (Bangladesh). Sci. Total Environ. 61(0):121-130. Khangarot, B.S. 1991. Toxicity of metals to a freshwater tubificid worm,Tubifex tubifex (Muller). Bull. Environ. Contam. Toxicol. 46(6):906-912. Kidwell, J.M., L.J. Phillips and G.F. Birchard. 1995. Comparative analyses of contaminant levels in bottom feeding and predatory fish using the national contaminant biomonitoring program data. Bull. Environ. Contam. Toxicol. 54(6):919-923. Kiffney, P. and A. Knight. 1990. The toxicity and bioaccumulation of selenate, selenite and seleno-l-methionine in the cyanobacterium Anabaena flos-aquae . Arch. Environ. Contam. Toxicol. 19(4):488-494. Kim, J.H., E. Birks and J.F. Heisinger. 1977. Protective action of selenium against mercury in northern creek chubs. Bull. Environ. Contam. Toxicol. 17:132-136. Kimball, G. Manuscript. The effects of lesser known metals and one organic to fathead minnows (Pimephales promelas) and Daphnia magna . (Available from C.E. Stephan, U.S. EPA, Duluth, MN.) King, K.A. and E. Cromartie. 1986. Mercury, cadmium, lead, and selenium in three waterbird species nesting in Galveston Bay, Texas, USA. Colonial Waterbirds 9(1):90-94. King, K.A., T.W. Custer and J.S. Quinn. 1991. Effects of Mercury, Selenium and Organochlorine Contaminants on Reproduction of Forster's Terns and Black Sk immers Nesting in a Contaminated Texas USA Bay. Arch. Environ. Contam. Toxicol. 20(1):32-40. King, K.A., T.W. Custer and D.A. Weaver. 1994. Reproductive success of barn swallows nesting near a selenium-contaminated lake in east Texas, USA. Environ. Pollut. 84(1):53-58. Kitamura, H. 1990. Relation between the toxicity of some toxicants to the aquatic animals ( Tanichthys albonubesand Neocaridina denticulata ) and the hardness of the test. Bull. Fac. Fish. Nagasaki Univ. Chodai Sui Kempo 67:13-19. Klauda, R.J. 1985a. Influence of delayed initial feeding on mortality of striped bass larvae exposed to arsenic and selenium. Am. Fish. Soc. Annu. Meeting No. 115:92-93. Klauda, R.J. 1985b. Acute and chronic effects of arsenic and selenium on the early life stages of striped bass. Report to Maryland Power Plant Siting Program, Maryland Department of Natural Resources. Publication JHU/APL PRPR-98. The Johns Hopkins University, Applied Physics Laboratory, Laurel, MD. Klaverkamp, J.F., D.A. Hodgins and A. Lutz. 1983a. Selenite toxicity and mercury-selenium interactions in juvenile fish. Arch. Environ. Contam. Toxicol. 12:405-413. Klaverkamp, J.F., W.A. Macdonald, W.R. Lillie and A. Lutz. 1983b. Joint toxicity of mercury and selenium in salmonid eggs. Arch. Environ. Contam. Toxicol. 12:415-419. Kleinow, K.M. 1984. The uptake, disposition, and elimination of selenate, selenite and selenomethionine in

### Draft November 12, 2004 Ref-47 the fathead minnow (

Pimephales promelas). Ph.D. thesis. University of Wisoconsin-Milwaukee, Milwaukee, WI. Available from: University Microfilms, Ann Arbor, MI. Order NO. 85-09260. Kleinow, K.M. and A.S. Brooks. 1986a. Selenium compounds in the fathead minnow ( Pimephales promelas) - I. Uptake, distribution, and elimination of orally administered selenate, selenite, and 1- selenomethionine. Comp. Biochem. Physiol. 83C:61-69. Kleinow, K.M. and A.S. Brooks. 1986b. Selenium compounds in the fathead minnow ( Pimephales promelas) - II. Quantitative approach to gastrointestinal absorption, routes of elimination and influence of dietary pretreatment. Comp. Biochem. Physiol. 83C:71-76. Klusek, C.S., M. Heit and S. Hodgkiss. 1993. Trace element concentrations in the soft tissue of transplanted freshwater mussels near a coal-fired power plant. In: Trace Elem. Coal Coal Combust. Residues. Keefer, R.F. and K.S. Sajwan (Eds). Lewis: Boca Raton, Fla. pp. 59-95.Koeman, J.H., W.H.M. Peeters, C.H.M. Koudstaal-Hol, P.S. Tijoe and J.J.M. de Goeij. 1973. Mercuryselenium correlations in marine mammals. Nature 245:385-386. Koenker R. 1994. Confidence intervals for regression quantiles. In Mandl P, Hu š ková M, eds, Asymptotic statistics: Proceedings of the Fifth Prague Symposium. Physica-Verlag, Heidelberg, Germany, pp 349-359. Koenker, R., and K. F. Hallock. 2001. Quantile regression: An introduction. J. Econom. Perspect. 15:143- 156. Koenker, R., and J. Gilbert Bassett. 1978. Regression Quantiles. Econometrica. 46:33-50. Koenker, R., and J. A. F. Machado. 1999. Goodness of fit and related inference processes for quantile regression. J. Am. Stat. Assoc. 94:1296-1310. Koenker R, Portnoy S. 1996. Quantile regression. Working Paper 97-0100. University of Illinois at Urbana-Champaign, College of Commerce and Business Administration, Office of Research, Urbana-Champaign. Koh, T.S. and M.J. Harper. 1988. Lead-poisoning in black swans, Cygnus atratus , exposed to spent lead shot at Bool Lagoon Game Reserve, South Australia. Aust. Wild. Res. 15(4):395-404. Koike, Y., Y. Nakaguchi, K. Hiraki, T. Takeuchi, T. Kokubo, and T. Ishimaru. 1993. Species and concentrations of selenium and nutrients in Tanabe Bay during red tide due to Gymnodinium nagasakiense

. J. Oceanog. 49(6):641-656. Kovacs, M., I. Nyary, L. Toth. 1984. The microelement content of some submerged and floating aquatic plants. Acta Botancia Hungarica 30(1-2):173-186. Kralj, N. and A. Stunja. 1994. Effects of selenium, lead and magnesium on the activity of hydrolytic enzymes in kidneys of the carp ( Cyprinus carpioL.). Period. Biol. 96(4):496-498. Kramer, K.J.M., H.A. Jenner and D. DeZwart. 1989. The valve movement response of mussels: a tool in

### Draft November 12, 2004 Ref-48 biological monitoring. Hydrobiol. 188-189:433-443. Krishnaja, A.P., M.S. Rege and A.G. Joshi. 1987. Toxic effects of certain heavy metals (mercury, cadmium, lead, arsenic and selenium) on the intertidal crab Scylla serrata . Mar. Environ. Res. 21(2):109-120. Krizkova, L., L. Horniak, S. Slavikova and L. Ebringer. 1996. Protective effect of sodium selenite on ofloxacin-induced loss of chloroplast DNA in Euglena gracilis . Fol. Microbiol. 41(4):329-332. Krogh, M. and P. Scanes. 1997. Organochlorine compound and trace metal contaminants in fish near Sydney's ocean outfalls. Mar. Pollut. Bull. 33(7-12):213-225. Krushevska, A., A. Lasztity, M. Kotrebai and R.M. Barnes. 1996. Addition of tertiary amines in the semiquantitative, multi-element inductively coupled plasma mass spectrometric analysis of biological materials. J. Anal. At. Spectrom. 11(5):343-352. Kruuk, H. and J.W.H. Conroy. 1991. Mortality of Otters Lutra lutra in Shetland Scotland UK. J. Appl. Ecol. 28(1):83-94. Kuehl, D.W. and R. Haebler. 1995. Organochlorine, organobromine, metal, and selenium residues in bottlenose dolphins (

Tursiops truncatus ) collected during an unusual mortality event in the Gulf of Mexico, 1990. Arch. Environ. Contam. Toxicol. 28(4):494-499. Kuehl, D.W., R. Haebler and C. Potter. 1994. Coplanar PCB and metal residues in dolphins from the U.S. Atlantic Coast including Atlantic bottlenose obtained during the 1987-88 mass mortality. Chemosphere 28(6):1245-1253. Kuliev, G.K. 1984. Effect of selenium on nutria growth and reproductive capacity. Sel'Skokhozyaistvennaya Biologiya 0(10):121-122. Kumar, H.D. 1964. Adaption of a blue-green alga to sodium selenate and chloramphenicol. Cell Physiol. 5:465. Kumar, H.D., and G. Prakash. 1971. Toxicity of selenium to the blue-green algae, Anacystis nidulansand Anabaena variabilis . Ann. Bot. (Lond.) 35:697-705. Kuss, S., S. Thakral and J. Behjan. 1995. Arroyo Simi characterization: A multi-purpose stream study to facilitate site specific permit requirements. Proc. Water Environ. Fed. Annu. Conf. Expo., 68th, Volume 4. Water Environment Federation: Alexandria, Va. pp. 307-317. Lahermo, G., Alfthan, and Wang. 1998. Selenium and Arsenic in the Environment in Finland. Journal of Environmental Pathology, Toxicology and Oncology. 17:205- . Lakshmanan, P.T. and J. Stephen. 1994. Trace metals in cephalopod mollusks - a unique phenomenon in metal accumulation. Nutr. Bioact. Subst. Aquat. Org., Pap. Symp., Meeting Date 1993. Devadasan, K. (Ed). Soc. Fish. Technol. (India): Cochin, India. pp. 254-265.

### Draft November 12, 2004 Ref-49 Lalitha, K. and P. Rani. 1995. Mitochondrial selenium-75 uptake and regulation revealed by kinetic analysis. Biol. Trace Element Res. 49(1):21-42. Lalitha, K., P. Rani and V. Narayanaswami. 1994. Metabolic relevance of selenium in the insect Corcyra cephalonica : Uptake of 75Se and subcellular distribution. Biol. Trace Element Res. 41(3):217-233. LamLeung, S.Y., V.K.W. Cheng and Y.W. Lam. 1991. Application of a microwave oven for drying and nitric acid extraction of mercury and selenium from fish tissue. Analyst (London) 116(9):957-959. Lan, W.G., M.K. Wong and Y.M. Sin. 1994a. Microwave digestion of fish tissue for selenium determination by differential pulsed polarography. Talanta 41(1):53-58. Lan, W.G., M.K. Wong, Y.M. Sin. 1994b. Comparison of four microwave digestion methods for the determination of selenium in fish tissue by using hydride generation atomic absorption spectrometry. Talanta 41(2):195-200. Lan, W.G., M.K. Wong, N. Chen, and Y.M. Sin. 1995. Effect of combined copper, zinc, chromium and selenium by orthogonal array design on alkaline phosphatase activity in liver of the red sea bream, Chrysophrys major . Aquaculture 131(3-4):219-230. Landau, M., R.H. Pierce, L.D. Williams and D.R. Norris. 1985. Contamination and growth of the shrimp, Penaeus stylirostrisStimpson, cultured in a seawater/wastewater aquaculture system. Bull. Environ. Contam. Toxicol. 35(4):537-45. Langlois, C. and R. Langis. 1995. Presence of airborne contaminants in the wildlife of northern Quebec. Sci. Total Environ. 160-161(0):391-402. Larsen, E.H. and S. Stuerup. 1994. Carbon-enhanced inductively coupled plasma mass spectrometric detection of arsenic and selenium and its application to arsenic speciation. J. Anal. At. Spectrom. 9(10):1099-1105. Larsen, E.H., G.A. Pedersen, and J.W. McLaren. 1997. Characterization of National Food Agency shrimp and plaice reference materials for trace elements and arsenic species by atomic and mass spectrometric techniques. J. Anal. At. Spectrom. 12(9):963-968. Larsen, L.F. and P. Bjerregaard. 1995. The effect of selenium on the handling of mercury in the shore crab Carcinus maenas . Mar. Pollut. Bull.31(1-3):78-83. Lauchli, A. 1993. Selenium in plants: Uptake, functions, and environmental toxicity. Botanica Acta 106(6):455-468. Law, R.J., R.L. Stringer, C.R. Allchin and B.R. Jones. 1996. Metals and organochlorines in sperm whales (Physeter macrocephalus ) stranded around the North Sea during the 1994/1995 winter. Mar. Pollut. Bull. 32(1):72-77. LeBlanc, G.A. 1980. Acute toxicity of priority pollutants to water flea ( Daphnia magna). Bull. Environ. Contam. Toxicol. 24:684-691.

### Draft November 12, 2004 Ref-50 LeBlanc, G.A. 1984. Interspecies relationships in acute toxicity of chemicals to aquatic organisms. Environ. Toxicol. Chem. 3:47-60. Lee, B.G. and N.S. Fisher. 1992a. Decomposition and release of elements from zooplankton debris. Mar. Ecol. Prog. Ser. 88(2-3):117-128. Lee, B.G. and N.S. Fisher. 1992b. Degradation and elemental release rates from phytoplankton debris and their geochemical implications. Limnol. Oceanogr. 37(7):1345-1360. Lee, B.G. and N.S. Fisher. 1993. Release rates of trace elements and protein from decomposing planktonic debris. 1. Phytoplankton debris. J. Mar. Res. 51(2):391-421. Leighton, F.A. and G. Wobeser. 1994. Salinity and selenium content in western Canadian wetlands. Wildl. Soc. Bull. 22(1):111-116. Leland, H.V. and B.C. Scudder. 1990. Trace elements in Corbicula flumineafrom the San Joaquin River, California. Sci. Total Environ. 97-98:641-672. Lemaire, P., A. Viarengo, L. Canesi, and D.R. Livingstone. 1993. Pro-oxidant and antioxidant processes in gas gland and other tissues of cod ( Gadus morhua ). J. Comp. Physiol. -B, Biochem., System., Environ. Physiol. 163(6):477-486. Lemly, A.D. 1982. Response of juvenile centrarchids to sublethal concentrations of waterbourne selenium. I. Uptake, tissue distribution, and retention. Aquat. Toxicol. 2:235-252. Lemly, A.D. 1983. A simple activity quotient for detecting pollution-induced stress in fishes. Environ. Technol. Lett. 4(4):173-178. Lemly, A.D. 1985a. Toxicology of selenium in a freshwater reservoir: Implications for environmental hazard evalutaion and safety. Ecotoxicol. Environ. Safety. 10:314-338. Lemly, A.D. 1985b. Ecological basis for regulating aquatic emissions from the power industry: The case with selenium. Regul. Toxicol. Pharmacol. 5:465-486. Lemly, A.D. 1993a. Metabolic stress during winter increases the toxicity of selenium to fish. Aquat. Toxicol. (Amsterdam) 27(1-2):133-158. Lemly, A.D. 1993b. Teratogenic effects of selenium in natural populations of freshwater fish. Ecotoxicol. Environ. Safety 26(2):181-204. Lemly, A.D. 1993c. Guidelines for evaluating selenium data from aquatic monitoring and assessment studies. Environ. Monitor. Assess. 28(1):83-100. Lemly, A.D. 1994. Agriculture and wildlife: Ecological implications of subsurface irrigation drainage. J. Arid Environ. 28(2):85-94. Lemly, A.D. 1995. A protocol for aquatic hazard assessment of selenium. Ecotoxicol. Environ. Safety

### Draft November 12, 2004 Ref-51 32(3):280-288. Lemly, A.D. 1996a. Assessing the toxic threat of selenium to fish and aquatic birds. Environ. Monitor. Assess. 43(1):19-35. Lemly, A.D. 1996b. Winter stress syndrome: an important consideration for hazard assessment of aquatic pollutants. Ecotoxicol. Environ. Safety. 34(3):223-227. Lemly, A.D. 1997a. Ecosystem recovery following selenium contamination in a freshwater reservoir. Ecotoxicol. Environ. Safety. 36(3):275-281. Lemly, A.D. 1997b. A teratogenic deformity index for evaluating impacts of selenium on fish populations. Ecotoxicol. Environ. Safety. 37:259-266. Lemly, A.D. 1997c. Environmental hazard of selenium in the Animas La Plata water development project. Ecotoxicol. Environ. Safety. 37:92-96. Lemly, A.D. 1997d. Environmental implications of excessive selenium: A review. Biomed. Environ. Sci. 10(4):415-435. Lemly, A.D. 1998. A position paper on selenium in ecotoxicology: a procedure for deriving site-specific water quality criteria. Ecotoxicol. Environ. Safety. 39:1-9. Lemly, A.D. and G.J. Smith. 1987. Aquatic cycling of selenium: Implications for fish and wildlife. U.S. Dept. of the Interior, U.S. Fish and Wildlife Service, Fish and Wildlife Leaflet 12. 10 pp. Leonzio, C., C. Fossi and S. Focardi. 1986. Lead, mercury, cadmium and selenium in two species of gull feeding on inland dumps, and in marine areas. Sci. Total Environ. 57(0):121-128. Leonzio, C., M. Lambertini, A. Massi, S. Focardi, and C. Fossi. 1989. An assessment of pollutants in eggs of Audouin's gull (

Larus audouinii ), a rare species of the Mediterranean Sea. Sci. Total Environ. 78(0):13-22. Leonzio, C., S. Focardi and C. Fossi. 1992. Heavy Metals and Selenium in Stranded Dolphins of the Northern Tyrrhenian Northwest Mediterranean. Sci. Total Environ. 119:77-84. Leskinen, J., O.V. Lindqvist, J. Lehto, Jari and P. Koivistoinen. 1986. Selenium and mercury contents in Northern pike (

Esox lucius , L.) of Finnish man-made and natural lakes. Vesientutkimuslaitoksen Julk. 65:72-9. Levander, O.A. 1977. Metabolic interrelationships between arsenic and selenium. Environ. Health Perspect. 19:159-164. Li, H., H. Nagasawa and K. Matsumoto. 1996. Graphite-furnace atomic absorption spectrometry of organomercury and organoselenium in extracts of biological samples with an organopalladium matrix modifier. Anal. Sci. 12(2):215-218.

### Draft November 12, 2004 Ref-52 Lie, O., E. Lied, A. Maage, L.R. Njaa and K. Sandnes. 1994. Nutrient content of fish and shellfish. Fiskeridir. Skr., Ser. Ernaer. 6(2):83-105. Lim, C. and D.M. Akiyama. 1995. Nutrient requirements of penaeid shrimp. In: Nutr. Util. Technol. Aquacult. 60-73. Lim, C.E. and D.J. Sessa (Eds). AOCS Press: Champaign, Ill. pp. 60-73. Lindstrom, K. 1984. Selenium and algal growth. In: Proceedings of the third international symposium on industrial uses of selenium and tellurium. Selenium-Tellurium Development Association, Darien, CT. pp. 441-469. Lindstrom, K. 1985. Selenium requirement of the dinoflagellate Peridinopsis borgei(Lemm). Int. Rev. Gesamten Hydrobiol. 70:77-85. Lindstrom, K. 1991. Nutrient Requirements of the Dinoflagellate Peridinium gatunense . J. Phycol. 27(2):207-219. Lipinski, N.G., P.M. Huang, W.K. Liaw and U.T. Hammer. 1986. The effects of chemical treatments on the retention and redox reactions of selenium by selected freshwater sediments. In: Proceedings of the twelfth annual aquatic toxicity workshop. Ozburn, G.W. (Ed.). Canadian Technical Report of Fisheries and Aquatic Sciences No. 1462. Department of Fisheries and Oceans, Ottawa, Ontario, Canada. pp. 166- 184. Liu, D.L., Y. P. Yang, M. H. Hu, P. J. Harrison and N. M. Price. 1987. Selenium content of marine food chain organisms from the coast of China. Mar. Environ. Res. 22(2): 151-165. Liu, Y.F., R.H. Tang, Q.X. Zhang, J.Y. Shi, X.M. Li, Z.Q. Liu and W. Zhao. 1986. Stimulation of cell growth of Tetrahymena pyriformisand Chlamydomonas reinhardtiiby trace elements. Biol. Trace Elem. Res. 9(2):89-99. Livingston, R.J., G.F. Brendel and D.A. Bruzek. 1991. Coal ash artificial reef demonstration. Proc. - Int. Ash Use Symp., 9th, Volume GS-7162, Vol. 2, 50/1-50/9. Electr. Power Res. Inst.: Palo Alto, Calif. Livingstone, D.R., F. Lips, P. Garcia Martinez and R.K. Pipe. 1992. Antioxidant Enzymes in the Digestive Gland of the Common Mussel Mytilus edulis . Mar. Biol. (Berlin) 112(2):265-276. Lizama, L.C., L.R. McDowell, and J.E. Marion. 1989. Utilization of aquatic plantsElodea canadensisand Hydrilla verticillatain laying hen diets: II. Macrominerals and microminerals. Nutr. Rep. Int. 39(3):521-536. Lobel, P.B., S.P. Belkhode, S.E. Jackson and H.P. Longerich. 1989. A universal method for quantifying and comparing the residual variability of element concentrations in biological tissues using elements in the mussel Mytilus edulisas a model. Mar. Biol. (Berlin) 102(4):513-518. Lobel, P.B., S.P. Belkhode, S.E. Jackson and H.P. Longerich. 1990. Recent taxonomic discoveries concerning the mussel

Mytilus : implications for biomonitoring. Arch. Environ. Contam. Toxicol. 19(4):508-512. Lobel, P.B., H.P. Longerich, S.E. Jackson, S.P. Belkhode. 1991. A major factor contributing to the high

### Draft November 12, 2004 Ref-53 degree of unexplained variability of some elements concentrations in biological tissue: 27 elements in 5 organs of the mussel Mytilusas a model. Arch. Environ. Contam. Toxicol. 21(1):118-125. Lobel, P.B., C.D. Bajdik, S.P. Belkhode, S.E. Jackson and H.P. Longerich. 1992a. Improved protocol for collecting mussel watch specimens taking into account sex size condition shell shape and chronological age. Arch. Environ. Contam. Toxicol. 21(3):409-414. Lobel, P.B., S.P. Belkhode, C. Bajdik, W.E. Jackson and H.P. Longerich. 1992b. General characteristics of the frequency distributions of element concentrations and of interelemental correlations in aquatic organisms. Mar. Environ. Res. 33(2):111-126. Lohner, T.W., R.J. Reash, V.E. Willet and L.A. Rose. 2001. Assessment of tolerant sunfish populations (Lepomissp.) Inhabiting selenium-laden coal ash effluents, 1. hematological and population level assessment. Ecotoxicol. Environ. Safety 50:203-216. Lohner, T.W., R.J. Reash, V.E. Willet and L.A. Rose. 2001. Assessment of tolerant sunfish populations (Lepomissp.) Inhabiting selenium-laden coal ash effluents, tissue biochemistry evaluation. Ecotoxicol. Environ. Safety 50:217-224. Lohner, T.W., R.J. Reash, V.E. Willet and L.A. Rose. 2001. Assessment of tolerant sunfish populations (Lepomis sp.) Inhabiting selenium-laden coal ash effluents, 3.serum chemistry and fish health indicators. Ecotoxicol. Environ. Safety 50:225-232. Lonzarich, D. G., T.E. Harvey and J.E. Takekawa. 1992. Trace element and organochlorine concentrations in California Clapper Rail Rallus longirostris obsoletus Eggs. Arch. Environ. Contam. Toxicol. 23(2):147-153. Lorentzen, M., A. Maage and K. Julshamn. 1994. Effects of dietary selenite or selenomethionine on tissue selenium levels of Atlantic salmon ( Salmo salar). Aquaculture 121(4):359-367. Losi, M.E. and W.T. Frankenberger, Jr. 1997. Reduction of selenium oxyanions by Enterobacter cloacae strain SLD1a-1: Reduction of selenate to selenite. Environ. Toxicol. Chem. 16(9):1851-1858. Lourdes, M., A. Cuvin-Aralar and R.W. Furness. 1990. Tissue distribution of mercury and selenium in minnows, Phoxinus phoxinus . Bull. Environ. Contam. Toxicol. 45(5):775-782. Low, K.W. and Y.M. Sin. 1995. In vitro effect of mercuric chloride and sodium selenite on chemiluminescent response of pronephros cells isolated from tilapia, Oreochromis aureus . Bull. Environ. Contam. Toxicol. 55(6):909-915. Low, K.W. and Y.M. Sin. 1996. In vivo and in vitro effects of mercuric chloride and sodium selenite on some non-specific immune responses of blue gourami, Trichogaster trichopterus(Pallus). Fish Shellfish Immunol. 6(5):351-362. Lowe, T.P., T.W. May, W.G. Brumbaugh and D.A. Kane. 1985. National contaminant biomonitoring program: Concentrations of seven elements in freshwater fish, 1978-1981. Arch. Environ. Contam. Toxicol. 14:363-388.

### Draft November 12, 2004 Ref-54 Lucas, H.F., Jr., D.N. Edgington and P.J. Colby. 1970. Concentrations of trace elements in Great Lakes fishes. J. Fish. Res. Board Can. 27:677-684. Lunde, G. 1973. The presence of lipid-soluble selenium compounds in marine oils. Biochem. Biophys. Acta 304:76-80. Lundquist, T.J., F.B. Green, R.B. Tresan, R.D. Newman, W. J. Oswald and M.B. Gerhardt. 1994. The algal-bacterial selenium removal system: mechanisms and field study. In: Selenium Environ. Frankenberger, W.T., Jr. and S. Benson (Eds). Dekker: New York, N. J. pp. 251-278. Luoma, S.N. and D.J.H. Phillips. 1988. Distribution, variability, and impacts of trace elements in San Francisco Bay. Mar. Pollut. Bull. 19(9):413-25. Luoma, S.N., C. Johns, N.S. Fisher, N.A. Steinberg, R.S. Oremland and J.R. Reinfelder. 1992. Determination of selenium bioavailability to a benthic bivalve from particulate and solute pathways. Environ. Sci. Technol. 26(3):485-491. Lussier, S.M. 1986. U.S. EPA, Narraganesett, RI. (Memorandum to D.J. Hansen, U.S. EPA, Narragansett, RI.) Luten, J.B., et al. 1980. Mercury and selenium in marine and freshwater fish. J. Food Sci. 45:416. Lyle, J.M. 1986. Mercury and selenium concentrations in sharks from Northern Australian waters. Aust. J. Mar. Freshwater Res. 37(3):309-322. Lytle, T.F. and J.S. Lytle. 1982. Heavy metals in oysters and clams of St. Louis Bay, Mississippi. Bull. Environ. Contam. Toxicol. 29:50-57. Maage, A. and R. Waagboe. 1990. Zinc and selenium in tissues of young Atlantic salmon Salmo salar

fed diets containing different lipid sources at two levels of vitamin E. Fiskeridir. Skr., Ser. Ernaer. 3(2):21-29. Maage, A., H. Sveier and K. Julshamn. 1989. A comparison of growth rate and trace element accumulation in Atlantic salmon (

Salmo salar) fry fed four different commercial diets. Aquaculture 79(1-4):267-273. MacFarlane, R.D., G.L. Bullock and J.J.A. McLaughlin. 1986. Effects of five metals on susceptibility of striped bass to Flexibacter columnaris . Trans. Am. Fish Soc. 115:227-231. MacKay, N.J., M.N. Kazacos, R.J. Williams and M.I. Leedow. 1975. Selenium and heavy metals in black marlin. Mar. Pollut. Bull. 6:57-61. Mackey, E.A., P.R. Becker, R. Demiralp, R.R. Greenberg, B.J Koster, and S.A. Wise. 1996. Bioaccumulation of vanadium and other trace metals in livers of Alaskan cetaceans and pinnipeds. Arch. Environ. Contam. Toxicol. 30(4):503-512. MacPhee, C. and R. Ruelle. 1969. Lethal effects of 1888 chemicals upon four species of fish from Western North America. Univ. of Idaho Forest, Wildl. Range Exp. Station Bull. No. 3, Moscow, ID, 112 p.

### Draft November 12, 2004 Ref-55 Magos, L. and M. Webb. 1980. The interactions of selenium with cadmium and mercury. Crit. Rev. Toxicol. 8:1-42.
Mahan, C.A., V. Majidi and J.A. Holcombe. 1989. Evaluation of the metal uptake of several algae strains in a multicomponent matrix utilizing inductively coupled plasma emission spectrometry. Anal. Chem. 61(6):624-7. Maher, W.A. 1985a. Characteristics of selenium in marine animals. Mar. Pollut. Bull. 16:33-34. Maher, W.A. 1985b. Selenium in macroalgae. Bot. Mar. 28(7):269-73. Maher, W.A. 1987. Distribution of selenium in marine animals: relationship to diet. Comp. Biochem. Physiol. C: Comp. Pharmacol. Toxicol. 86C(1):131-3. Maher, W., S. Baldwin, M. Deaker and M. Irving. 1992. Characteristics of selenium in Australian marine biota. Appl. Organomet. Chem. 6(2):103-112. Maher, W., M. Deaker, D. Jolley, F. Krikowa and B. Roberts. 1997. Selenium occurrence, distribution and speciation in the cockle

Anadara trapeziaand the mullet Mugil cephalus . Appl. Organomet. Chem. 11(4):313-326. Maier, K.J. 1990. The toxicity and bioaccumulation of selenium and boron to Daphnia magnaand Chironomus decorus . 191 pp. Avail. Univ. Microfilms Int., Order No. DA9102095 From: Diss. Abstr. Int. B 1991, 51(8):3656. Maier, K.J. and A.W. Knight. 1993. Comparative acute toxicity and bioconcentration of selenium by the midge Chironomus decorusexposed to selenate, selenite and seleno-dl-methionine. Arch. Environ. Contam. Toxicol. 25(3):365-370. Maier, K.J., C.G. Foe and A.W. Knight. 1993. Comparative toxicity of selenate, selenite, seleno-dl-methionine and seleno-dl-cystine to Daphnia magna . Environ. Toxicol. Chem. 12(4):755-763. Malarvizhi, K. and M.V. Usharani. 1994. Effect of sodium selenite on the cytological effects of methyl parathion in the root meristems of Allium cepa . J. Environ. Biol. 15(3):193-198. Malchow, D.E., A.W. Knight and K.J. Maier. 1995. Bioaccumulation and toxicity of selenium in Chironomus decoruslarvae fed a diet of seleniferous Selenastrum capricornutum . Arch. Environ. Contam. Toxicol. 29(1):104-109. Mancini, J.L. 1983. A method for calculating effects, on aquatic organisms, of time varying concentrations. Water Res. 17:1355-1362. Mann, H. and W.S. Fyfe. 1988. Biogeochemical cycling of the elements in some fresh water algae from gold and uranium mining districts. Biorecovery, 1(1):3-26. Mann, H., W.S. Fyfe and R. Kerrich. 1988. The chemical content of algae and waters: bioconcentration.

### Draft November 12, 2004 Ref-56 Toxic. Assess. 3(1):1-16. Manoharan, A. and V. Prabakaran. 1994. Acute toxicity and genotoxic effect of chromium and selenium on the common loach,

Lepidocephalichthyes thermalis(Bleeker). Geobios (Jodhpur) 21(1):44-46. Marcogliese, D.J., G.W. Esch and R.V. Dimock Jr. 1989. Long-term comparison of zooplankton communities between thermally-altered and ambient areas of a North Carolina Cooling Reservoir USA. J. Elisha Mitchell Sci. Soc. 105(1):1-13. Marcogliese, D.J., G.W. Esch and R.V. Dimock, Jr. 1992. Alterations in zooplankton community structure after selenium-induced replacement of a fish community a natural whole-lake experiment. Hydrobiol. 242(1):19-32. Martin, J.L. 1973. Selenium assimilation in animals. In: Organic selenium compounds: Their chemistry and biology. Klayman, D.L. and W.H.H. Gunther (Eds.). Wiley-Interscience, New York, NY. pp. 663-691. Martin, M., K.E. Osborn, P. Billig and N. Glickstein. 1981. Toxicities of ten metals to Crassostrea gigas and Mytilus edulisembryos and Cancer magister larvae. Mar. Pollut. Bull. 12:305-308. Marvin, C.H. and E.T. Howell. 1997. Contaminant burdens in sediments colonized by Dreissena at two nearshore sites in the lower Great Lakes. Zebra Mussels Aquat. Nuisance Species, [Proc. Int. Zebra Mussel Other Aquat. Nuisance Species Conf.], 6th, Meeting Date 1996, 209-224. Editor(s): D'Itri, Frank M. Mason, R.P., J.-M.Laporte, and S. Andres. 2000. Factors controlling the bioaccumulation of mercury, arsenic, selenium, and cadmium by freshwater invertebrates and fish. Archives Environ. Contamin. Toxicol. 38:283-297. Masuzawa, T., M. Koyama and M. Terazaki. 1988. A regularity in trace element contents of marine zooplankton species. Mar. Biol. (Berlin) 97(4):587-91 Matsumoto, K. 1991. Speciation and determination of selenium and mercury accumulated in a dolphin liver. ACS Symp. Ser., 445:278-289. Mattice, J.S., D.B. Povella and R.W. Brocksen. 1997. Sediment-water interactions affect assessments of metals discharges at electric utilities. Water Air Soil Pollut. 99:187-199. Mauk, R.J. and M.L. Brown. 1999. Acute toxicity of sodium selenite to juvenile walleye. Bull. Environ. Contam. Toxicol. 63: 188-194. Maven, H., K.S. Rao, W. Benko, K. Alam, M.E. Huber, T. Rali and I. Burrows. 1995. Fatty acid and mineral composition of

Papua New Guinea echinoderms. Fish. Technol. 32(1):50-52. May, T.W. and D.A. Kane. 1984. Matrix-dependent instability of selenium(IV) stored in teflon containers. Anal. Chim. Acta 161:387-391. May, T.W. and G.L. McKinney. 1981. Cadmium, lead, mercury, arsenic, and selenium concentrations in

### Draft November 12, 2004 Ref-57 freshwater fish, 1976-77 - National Pesticide Monitoring Program. Pestic. Monit. J. 15:14-38. May, T.W., M.J. Walthier, J.D. Petty, J.F. Fairchild, J. Lucero, M. Delvaux, J. Manring, M. Arbruster and D. Hartman. 2001. An evaluation of selenium concentration in water, sediment, invertebrates, and fish from the Republican River Basin: 1997-1999. Environ. Monit. Assess. 72: 179-206. Mayer, F.L., Jr. and M.R. Ellersieck. 1986. Manual of acute toxicity: Interpretation and data base for 410 chemicals and 66 species of freshwater animals. Resource Publication No. 160. U.S. Fish and Wildlife Service, Washington, DC. p. 438. Mayer, F.L., Jr., K.S. Mayer and M.R. Ellersieck. 1986. Relation of survival to other endpoints in chronic toxicity tests with fish. Environ. Toxicol. Chem. 5(8):737-48. McCloskey, J.T. and M.C. Newman. 1995. Sediment preference in the asiatic clam ( Corbicula fluminea ) and viviparid snail (

Campeloma decisum) as a response to low-level metal and metalloid contamination. Arch. Environ. Contam. Toxicol. 28(2):195-202. McCloskey, J.T., M.C. Newman and P.M. Dixon. 1995. Effect of metal and metalloid contaminated sediment on the spatial distribution of asiatic clams ( Corbicula fluminea ). Arch. Environ. Contam. Toxicol. 28(2):203-208. McCrea, R.C. and J.D. Fischer. 1986. Heavy metal and organochlorine contaminants in the five major Ontario rivers of the Hudson Bay Lowland. Water Pollut. Res. J. Can. 21(2):225-34. Mcdowell, L. R., D.J. Forrester, S.B. Linda, S.D. Wright, and N.S. Wilkinson. 1995. Selenium status of white-tailed deer in southern Florida. J. Wild. Dis. 31(2):205-211. McKee, J.E. and H.W. Wolf. 1963. Water quality criteria. 2nd ed. Publication No. 3-A. State Water Quality Control Board, Sacramento, CA. pp. 253-254. McKenzie-Parnell, J.M., T.E. Kjellstrom, R.P. Sharma and M.F. Robinson. 1988. Unusually high intake and fecal output of cadmium, and fecal output of other trace elements in New Zealand adults consuming dredge oysters. Environ. Res. 46(1):1-14. Mclean, C., A.G. Miskiewicz and E.A. Roberts. 1991. Effect of three primary treatment sewage outfalls on metal concentrations in the fishCheilodactylus fuscuscollected along the Coast of Sydney Australia. Mar. Pollut. Bull. 22(3):134-140. Meador, J.P., U. Varanasi, P.A. Robisch and S.-L. Chan. 1993. Toxic metals in pilot whales ( Globicephala melaena) from strandings in 1986 and 1990 on Cape Cod, Massachusetts. Can. J. Fish. Aquat. Sci. 50(12):2698-2706. Measures, C.I. and J.D. Burton. 1978. Behavior and speciation of dissolved selenium in estuarine waters. Nature 273:293-295. Mehrle, P.M., T.A. Haines, S. Hamilton, J.L. Ludke, F.L. Mayer and M.A. Ribick. 1982. Relationship between body contaminants and bone development in east-coast striped bass. Trans. Am. Fish. Soc.

### Draft November 12, 2004 Ref-58 111:231-241. Mehrle, P.M., L. Cleveland and D.R. Buckler. 1987. Chronic toxicity of an environmental contaminant mixture to young (or larval) striped bass. Water, Air, Soil Pollut. 35(1-2):107-18. Meltzer, H.M., K. Bibow, I.T. Paulsen, H.H. Mundal, G. Norheim and H. Holm. 1993. Different Bioavailability in Humans of Wheat and Fish Selenium as Measured by Blood Platelet Response to Increased Dietary Selenium. Biol. Trace Element Res. 36(3):229-241. Metcalfe-Smith, J.L. 1994. Influence of species and sex on metal residues in freshwater mussels (family Unionidae) from the St. Lawrence River, with implications for biomonitoring programs. Environ. Toxicol. Chem. 13(9):1433-1443. Metcalfe-Smith, J.L., J.C. Merriman and S.P. Batchelor. 1992. Relationships Between Concentrations of Metals in Sediment and Two Species of Freshwater Mussels in the Ottawa River. Water Pollut. Res. J. Can. 27(4):845-869. Metcalfe-Smith, J.L., R.H. Green and L.C. Grapentine. 1996. Influence of biological factors on concentrations of metals in the tissues of freshwater mussels ( Elliptio complanataand Lampsilis radiata ) from the St. Lawrence River. Can. J. Fish. Aquat. Sci. 53(1):205-219. Micallef, S. and P.A. Tyler. 1987. Preliminary observations of the interactions of mercury and selenium in Mytilus edulis . Mar. Pollut. Bull. 18(4):180-185. Micallef, S. and P.A.Tyler. 1989. Levels and interactions of selenium with group IIB metals in mussels from Swansea Bay, South Wales, U.K. Bull. Environ. Contam. Toxicol. 42(3):344-51. Micallef, S. and P.A. Tyler. 1990. Effect of Mercury and Selenium on the Gill Function of Mytilus edulis

. Mar. Pollut. Bull. 21(6):288-292. Michot, T.C., T.W. Custer, A.J. Nault, and C.A. Mitchell. 1994. Environmental contaminants in redheads wintering in coastal Louisiana and Texas. Arch. Environ. Contam. Toxicol. 26(4):425-434. Mikac, N., M. Picer, P. Stegnar, M. Tusek-Znidaric. 1985. Mercury distribution in a polluted marine area, ratio of total mercury, methylmercury and selenium in sediments, mussels and fish. Water Res. 19(11):1387-1392. Miles, A.K. and M.W. Tome. 1997. Spatial and temporal heterogeneity in metallic elements in industrialized aquatic bird habitats. Environ. Pollut. 95(1):75-84. Miller, J.J., B.J. Read, D.J.Wentz, and D.J. Heaney. 1996. Major and trace element content of shallow groundwater associated with dryland saline soils in southern Alberta. Water Qual. Res. J. Can. 31(1):101-117. Mills, E.L., E.F. Roseman, M. Rutzke, W.H. Gutenmann and D.J. Lisk. 1993. Contaminant and nutrient element levels in soft tissues of zebra and quagga mussels from waters of southern Lake Ontario. Chemosphere 27(8):1465-1473.

### Draft November 12, 2004 Ref-59 Milne, J.B. 1998. The Uptake and Metabolism of Inorganic Selenium Species. In: W.T. Frankenberger, Jr. and R.A. Engberg (eds.), Environmental Chemistry of Selenium. Marcel Dekker, New York. pp. 459-478. Minganti, V., F. Fiorentino, R. De Pellegrini and R.Capelli. 1994. Bioaccumulation of mercury in the Antarctic bony fish Pagothenia bernacchii. Int. J. Environ. Anal. Chem. 55(1-4):197-202. Minganti, V., R. Capelli, F. Fiorentino, R. De Pellegrini and M. Vacchi. 1995. Variations of mercury and selenium concentrations in Adamussium colbecki

and Pagothenia bernacchii from Terra Nova Bay (Antarctica) during a five year period. Int. J. Environ. Anal. Chem. 61(3):239-248. Misitano, D.A. and M.H. Schiewe. 1990. Effect of chemically contaminated marine sediment on naupliar production of the marine harpacticoid copepod, Tigriopus californicus . Bull. Environ. Contam. Toxicol. 44(4):636-642. Moede, A., R.W. Greene and D.F. Spencer. 1980. Effects of selenium on the growth and phosphorus uptake of Scenedesmus dimorphus and Anabaena cylindrica . Environ. Exp. Bot. 20:207-212. Moharram, Y.G., S.A. El-Sharnouby, E.K. Moustaffa and A. El-Soukkary. 1987. Mercury and selenium content in bouri (

Mugil cephalus ). Water Air Soil Pollut. 32:455-459. Moller, G. 1996. Biogeochemical interactions affecting hepatic trace element levels in aquatic birds. Environ. Toxicol. Chem. 15(7):1025-1033. Montagnese, C.M., F.A. Geneser and J.R. Krebs. 1993. Histochemical distribution of zinc in the brain of the Zebra Finch Taenopygia guttata . Anat. Embryol. 188(2):173-187. Moore, J.F. 1988. Selenium toxicosis in wild aquatic birds. J. Toxicol. Environ. Health 24(1):67-92. Mora, M.A. and D.W. Anderson. 1995. Selenium, boron, and heavy metals in birds from the Mexicali Valley, Baja California, Mexico. Bull. Environ. Contam. Toxicol. 54(2):198-206. Morera, M., C. Sanpera, S. Crespo, L. Jover, and X. Ruiz. 1997. Inter- and intraclutch variability in heavy metals and selenium levels in Audouin's gull eggs from the Ebro Delta, Spain. Arch. Environ. Contam. Toxicol. 33(1):71-75. Muir, D.C.G., R. Wagemann, N.P. Grift, R.J. Norstrom, M. Simon and J. Lien. 1988. Organochlorine chemical and heavy metal contaminants in white-beaked dolphins ( Lagenorhynchus albirostris ) and pilot whales ( Globicephala melaena) from the coast of Newfoundland, Canada. Arch. Environ. Contam. Toxicol. 17(5):613-630. Munawar, M. and M. Legner. 1993. Detection of metal toxicity using natural phytoplankton as test organisms in the Great Lakes. Water Pollut. Res. J. Can. 28(1):155-176. Munawar, M., I.F. Munawar, P.E. Ross and C.I. Mayfield. 1987. Differential sensitivity of natural phytoplankton size assemblages to metal mixture toxicity. Ergeb. Limnol. 25:123-39. Murata, H., T. Sakai, K. Yamauchi, T. Ito, T. Tsuda, T. Yoshida and M. Fukudome. 1996. In vivo lipid

### Draft November 12, 2004 Ref-60 peroxidation levels and antioxidant activities of cultured and wild yellowtail. Fish. Sci. 62(1):64-68. Muskett, C.J, R. Chan, J. Towner and R. Singleton. 1985. Assessment of the impact of heavy metals released from a fly ash lagoon on a commercial oyster bed. Heavy Met. Environ. Int. Conf. 5th, Volume 1, 661-3. Editor(s): Lekkas, Themistokles D. CEP Consult.: Edinburgh, UK. Mutanen, M., P. Koivistoinen, V.C. Morris and O.A. Levander. 1986. Nutritional availability to rats of selenium in four seafoods: Crab ( Callinectes sapidus ), oyster ( Crassostrea virginica), shrimp ( Penaeus duorarum) and Baltic herring ( Clupea harengus ). Br. J. Nutr. 55(2):219-226. Naddy, R.B., T.W. LaPoint, and S.J. Klaine. 1995. Toxicity of arsenic, molybdenum and selenium combinations to CerioDaphnia dubia . Environ. Toxicol. Chem. 14(2):329-336. Nadkarni, N.M. and R.B. Primack. 1989. The use of gamma spectrometry to measure within-plant nutrient allocation of a tank bromeliad Guzmania lingulata . Selbyana 11:22-25. Naftz, D.L., R.B. See and P. Ramirez. 1993. Selenium source identification and biogeochemical processes controlling selenium in surface water and biota, Kendrick Reclamation Project, Wyoming, U.S.A. Appl. Geochem. 8(2):115-126. Nakamoto, R.J. and T.J. Hassler. 1992. Selenium and other trace elements in bluegills from agricultural return flows in the San Joaquin Valley California. Arch. Environ. Contam. Toxicol. 22(1):88-98. Nakonieczny, M. 1993. Functional aspects of cadmium and selenium interactions in insect digestive tract: Enzyme studies. Sci. Total Environ. (Suppl. Part 1):573-583. Narasaki, H. 1985. Determination of arsenic and selenium in fat materials and petroleum products by oxygen bomb combustion and automated atomic absorption spectrometry with hydride generation. Anal. Chem. 57(13):2481-6. Narasaki, H. and J.Y. Cao. 1996. Determination of arsenic and selenium by hydride generation atomic absorption spectrometry using a gas-liquid separator and a dehydration trap. Microchem. J. 53(1):18-25. Nassos, P.A., J.R. Coats, R.L. Metcalf, D.D. Brown and L.G. Hansen. 1980. Model ecosystem, toxicity, and uptake evaluation of 75 

### Se-selenium. Bull. Environ. Contam. Toxicol. 24:752-758. National Academy of Sciences. 1976. Selenium. In: Medical and Biological Effects of Environmental Pollutants . Washington, DC. p.23. National Research Council. 1976. Selenium. PB-251318 or EPA-600/1-76-014. National Technical Information Service, Springfield, VA. Navarrete, M., L. Cabrera, N. Deschamps, N. Boscher, G. Revel, J.P. Meyer and A. Stampfler. 1990. Activation analysis of selenium in biological samples through selenium-75 and selenium-77m. J. Radioanal. Nuclear Chem. 145(6):445-452. Nelson, D.A., J.E. Miller and A. Calabrese. 1988. Effect of heavy metals on bay scallops, surf clams, and

### Draft November 12, 2004 Ref-61 blue mussels in acute and long-term exposures. Arch. Environ. Contam. Toxicol. 17(5):595-600. Neter, J., W. Wasserman, and M. H. Kutner. 1985. Applied linear statistical models. Irwin, Homewood, Illinois. Nettleton, J.A., W.H. Allen, Jr., L.V. Klatt, W.M.N. Ratnayake and R.G. Ackman. 1990. Nutrients and chemical residues in one- to two-pound Mississippi farm-raised channel catfish ( Ictalurus punctatus ). J. Food Sci. 55(4):954-958. Neuhierl, B. and A. Boeck. 1996. On the mechanism of selenium tolerance in selenium-accumulating plants: Purification and characterization of a specific selenocysteine methyltransferase from cultured cells of Astragalus bisulcatus . European J. Biochem. 239(1):235-238. Neuhold, J.M. 1987. The relationship of life history attributes to toxicant tolerance in fishes. Environ. Toxicol. Chem. 6(9):709-16. Nicola, R.M., R. Branchflower and D. Pierce. 1987. Chemical contaminants in bottomfish. J. Environ. Health 49(6):342-7. Nielsen, C.O. and R. Dietz. 1990. Distributional pattern of zinc, cadmium, mercury and selenium in livers of hooded seal Cystophora cristata . Biol. Trace Element Res. 24(1):61-72. Nielsen, G. and P. Bjerregaard. 1991. Interaction between accumulation of cadmium and selenium in the tissues of turbot Scophthalmus maximus . Aquat. Toxicol. (Amsterdam) 20(4):253-266. Nigro, M. 1994. Mercury and selenium localization in macrophages of the striped dolphin, Stenella coeruleoalba . J. Marine Biol. Assoc. U.K. 74(4):975-978. Nigro, M., E. Orlando and F. Regoli. 1992. Ultrastructural localization of metal binding sites in the kidney of the Antarctic Scallop Adamussium colbecki . Mar. Biol. (Berlin) 113(4):637-643. Niimi, A.J. and Q.N. LaHam. 1975. Selenium toxicity on the early life stages of zebrafish ( Brachydanio rerio ). J. Fish. Res. Board Can. 32:803-806. Niimi, A.J. and Q.N. LaHam. 1976. Relative toxicity of organic and inorganic compounds of selenium to newly hatched zebrafish ( Brachydanio rerio ). Can. J. Zool. 54:501-509. Norberg-King, T.J. 1989. An evaluation of the fathead minnow seven-day subchronic test for estimating chronic toxicity. Environ. Toxicol. Chem. 8(11):1075-1089. Norheim, G. 1987. Levels and interactions of heavy metals in sea birds from Svalbard and the Antarctic. Environ. Pollut. 47(2):83-94. Norheim, G. and B. Borch. Iohnsen. 1990. Chemical and Morphological Studies of Liver from Eider Somateria mollissimain Svalbard Arctic Ocean with Special Reference to the Distribution of Copper. J. Comp. Pathol. 102(4):457-466.

### Draft November 12, 2004 Ref-62 Norheim, G., F. Mehlum, C. Bech and M.T. Moksnes. 1991. Distribution of Selenium Binding Proteins in Liver from Two Species of Penguins from Bouvetoya South Atlantic Ocean. Polar Research 9(1):109-111. Norheim, G., J.U. Skaare and O. Wiig. 1992. Some Heavy Metals Essential Elements And Chlorinated Hydrocarbons In Polar Bear Ursus maritimusAt Svalbard. Environ. Pollut. 77(1):51-57. Norman, B., G. Nader, M. Oliver, R. Delmas, D. Drake and H. George. 1992. Effects of selenium supplementation in cattle on aquatic ecosystems in Northern California. J. Am. Vet. Med. Assoc. 201(6):869-872. Norrgren, L., T. Andersson, P.A. Bergqvist and I. Bjoerklund. 1993. Chemical, physiological and morphological studies of feral Baltic salmon ( Salmo salar) suffering from abnormal fry mortality. Environ. Toxicol. Chem. 12(11):2065-2075. Norstrom, R.J., R.E. Schweinsberg and B.T. Collins. 1986. Heavy metals and essential elements in livers of the polar bear ( Ursus maritimus ) in the Canadian Arctic. Sci. Total Environ. 48(3):195-212. North Carolina Department of Natural Resources and Community Development. 1986. North Carolina water quality standards documentation: The freshwater chemistry and toxicity of selenium with an emphasis on its effects in North Carolina. Report No. 86-02. Raleigh, NC. O'Brien, D.J., R.H. Poppenga and C.W. Ramm. 1995. An exploratory analysis of liver element relationships in a case series of common loons ( Gavia immer

). Prev. Vet. Med. 25(1):37-49. O'Connor, T.P. 1996. Trends in chemical concentrations in mussels and oysters collected along the US coast from 1986 to 1993. Mar. Environ. Res. 41(2):183-200. O'Shea, T.J., J.F. Moore and H.I. Kochman. 1984. Contaminant concentrations in manatees ( Trichechus manatus ) in Florida (USA). J. Wildl. Manage. 48(3):741-748. Ober, A.G., M. Gonzalez and I. Santa Maria. 1987. Heavy metals in molluscan, crustacean, and other commercially important Chilean marine coastal water species. Bull. Environ. Contam. Toxicol. 38(3):534-9. Oberbach, H. and W. Hartfiel. 1987. Effects of different alpha-tocopherol and selenium additions in ratios with high contents of polyene acids on rainbow trouts ( Salmo gairdneri , R.). Fett Wissenschaft Technologie 89(5):195-199. Oberbach, H. and W. Hartfiel. 1988. Investigations of the alpha-tocopherol and selenium requirement of rainbow trout (

Salmo gairdneri , R.) and pathological deficiency symptoms in case of rations which are rich in polyene acids. Fett Wissenschaft Technologie 90(3):97-101. Oberbach, H., V. Totovic, and W. Hartfiel. 1989. Effects of differently high oxidized fats in feed of rainbow trouts (

Salmo gairdneri , R.) on the need for vitamin E and selenium. Fett Wissenschaft Technologie 91(4):148-153. Oehlenschlager, J. 1997. Marine fish - a source for essential elements?!. Dev. Food Sci. 38:641-652.

### Draft November 12, 2004 Ref-63 Ogle, R.S. and A.W. Knight. 1989. Effects of elevated foodborne selenium on growth and reproduction of the fathead minnowPimephales promelas . Arch. of Environ. Contam. Toxicol. 18(6):795-803. Ogle, R.S. and A.W. Knight. 1996. Selenium bioaccumulation in aquatic ecosystems: 1. Effects of sulfate on the uptake and toxicity of selenate in Daphnia magna . Arch. Environ. Contam. Toxicol. 30(2):274-279. Ohlendorf, H.M. 1986. Aquatic birds and selenium in the San Joaquin Valley. In: Selenium and agricultural drainage: Implications for San Francisco Bay and the California environment. The Bay Institute of San Francisco, Tiburon, CA. pp. 15-24. Ohlendorf, H.M. and C.S. Harrison. 1986. Mercury, selenium, cadmium and organochlorines in eggs of three Hawaiian (USA) seabird species. Environ. Pollut. Series B Chemical and Physical 11(3):169-192. Ohlendorf, H.M. and K.C. Marois. 1990. Organochlorines and Selenium in California USA Night heron and Egret Eggs. Environ. Monitor. Assess. 15(1):91-104. Ohlendorf, H.M., D.J. Hoffman, M.K. Saiki and T.W. Aldrich. 1986a. Embryonic mortality and abnormalities of aquatic birds: Apparent impacts of selenium from irrigation drainwater. Sci. Total Environ. 52:49-63. Ohlendorf, H.M., R.W. Lowe, P.R. Kelly and T.E. Harvey. 1986b. Selenium and heavy metals in San Francisco Bay diving ducks. J. Wildl. Manage. 50:64-71. Ohlendorf, H.M., R.L. Hothem, T.W. Aldrich, and A.J. Krynitsky. 1987. Selenium contamination of the Grasslands, a major California (USA) waterfowl area. Sci. Total Environ. 66(0):169-184 Ohlendorf, H.M., R.L. Hothem and T.W. Aldrich. 1988a. Bioaccumulation of selenium by snakes and frogs in the San Joaquin Valley, California (USA). Copeia 1988(3):704-710. Ohlendorf, H.M., A.W. Kilness, J.L. Simmons, R.K. Stroud, D.J. Hoffman, and J.F. Moore. 1988b. Selenium toxicosis in wild aquatic birds. J. Toxicol. Environ. Health 24(1):67-92. Ohlendorf, H.M., R.L. Hothem and D. Welsh. 1989. Nest Success Cause-specific Nest Failure and Hatchability of Aquatic Birds at Selenium-contaminated Kesterson Reservoir and a Reference Site. Condor 91(4):787-796. Ohlendorf, H.M., R.L. Hothem, C.M. Bunck and K.C. Marois. 1990. Bioaccumulation of Selenium in Birds at Kesterson Reservoir California USA. Arch. Environ. Contam. Toxicol. 19(4):495-507. Ohlendorf, H.M., K.C. Marois, R.W. Lowe, T.E. Harvey and P.R. Kelly. 1991. Trace Elements and Organochlorines in Surf Scoters from San Francisco Bay 1985 California USA. Environ. Monitor. Assess. 18(2):105-122. Okasako, J. and S. Siegel. 1980. Mercury antagonists: Effects of sodium chloride and sulfur group (VIa) compounds on encystment of the brine shrimp Artemia . Water Air Soil Pollut. 14:235-240. Okazaki, R.K. and M.H. Panietz. 1981. Depuration of twelve trace metals in tissues of the oysters

### Draft November 12, 2004 Ref-64 Crassostrea gigasand C. virginica . Mar. Biol. (Berl.) 63:113-120. Oliver, M.N., G. Ros-McGauran, D.A. Jessup, B.B. Norman, and C.E. Franti. 1990. Selenium concentrations in blood of free-ranging mule deer in California. Trans. West Sec. Wildl. Soc. 26:80-90. Olson, D.L. and G.M. Christensen. 1980. Effects of water pollutants and other chemicals on fish acetylcholinesterase (in vitro). Environ. Res. 21:327-335. Olson, M.M. and D. Welsh. 1993. Selenium In Eared Grebe Embryos From Stewart Lake National Wildlife Refuge North Dakota. Prairie Naturalist 25(2):119-126. Olson, O.E., E.J. Novacek, E.I. Whitehead and I.S. Palmer. 1970. Investigations on selenium in wheat. Phytochem. 9:1181-1188. Oppenheimer, J.A., A.D. Eaton and P.H. Kreft. 1984. Speciation of selenium in groundwater. EPA-600/2- 84-190 or PB85-125979. National Technical Information Service, Springfield, VA. Opresko, D.M., B.E. Sample, and G.W. Suter. 1995. Toxicological Benchmarks for Wildlife: 1995 Revision. U.S. Dept. Energy Oak Ridge National Laboratory. ES/ER/TM-86/R2. Oremland, R.S., J.T. Hollibaugh, A.S. Maest, T.S. Presser, L.G. Miller, and C.W. Cullbertson. 1989. Selenate Reduction to Elemental Selenium by Anaerobic Bacteria in Sediments and Culture: Biogeochemical Significance of a Novel Sulfate-Independent Respiration, ” Applied and Environmental Microbiology . 55:2333- . Ornes, W.H., K.S. Sajwan, M.G. Dosskey and D.C. Adriano. 1991. Bioaccumulation of selenium by floating aquatic plants. Water, Air, Soil Pollut., 57-58:53-57. Ostapczuk, P., M. Burow, K. May, C. Mohl, M. Froning, B. Suessenbach, E. Waidmann and H. Emons. 1997. Mussels and algae as bioindicators for long-term tendencies of element pollution in marine ecosystems. Chemosphere 34(9/10):2049-2058. Ostroymora, I.N., ZH.I. Abramora and V.M. Amelyutin. 1985. Effect of phosphatides and selenium on the effectiveness of replacement of fish meal with protein products of microbiosynthesis in the food of carp, Cyprinus carpio , in warm waters. Voprosy Ikhtiologii 25(6):1017-1023. Ostroymora, I.N., Z.I. Abramora and V.M. Amelyutin. 1986. The influence of phosphatides and selenium on the efficacy of the substitution of protein products from microbiosynthesis for fish meal in feed for carp, Cyprinus carpioin warm waters. J. Ichthol. (Engl. Transl. Vopr. Ikhtiol.) 26(1):98-105. Overbaugh, J.M. and R. Fall. 1985. Characterization of a selenium-independent glutathione peroxidase from Euglena gracilisvar. bacillaris. Plant Physiol. (Bethesda) 77(2):437-442. Owsley, J.A. 1984. Acute and chronic effects of selenium-selenium on CerioDaphnia affinis . M.S. thesis. Vanderbilt University, Nashville, TN. Owsley, J.A. and D.E. McCauley. 1986. Effects of extended sublethal exposure to sodium selenite on

### Draft November 12, 2004 Ref-65 CerioDaphnia affinis . Bull. Environ. Contam. Toxicol. 36:876-880. Oyamada, N., G. Takahashi and M. Ishizaki. 1991. Methylation of inorganic selenium compounds by freshwater green algaeAnkistrodesmus sp. , Chlorella vulgarisand Selenastrum sp. Eisei Kagaku 37(2):83-88. Pagano, G, M. Cipollaro, G. Corsale, A. Esposito, E. Ragucci, G.G.Giordano and N.M. Trieff. 1986. The sea urchin: bioassay for the assessment of damage from environmental contaminants. ASTM Spec. Tech. Publ., 920(Community Toxic. Test.):66-92. Pakkala, I.S., W.H. Gutenmann, D.J. Lisk, G.E. Burdick and E.J. Harris. 1972. A survey of the selenium content of fish from 49 New York state waters. Pestic. Monit. J. 6:107-114. Pal, B.K., M.J.U. Ahmed, A.K. Chakrabarti and D. Chakraborty. 1997. Spectrofluorometric determinations of chromium, selenium and manganese in their mixtures and their application to environmental and biological samples. Indian J. Chem. Technol. 4(4):191-195. Palawski, D., J.B. Hunn and F.J. Dwyer. 1985. Sensitivity of young striped bass to organic and inorganic contaminants in fresh and saline watrs. Trans. Am. Fish. Soc. 114:748-753. Palawski, D.U., W.E. Jones, K. DuBois and J.C. Malloy. 1991. Contaminant biomonitoring at the Benton Lake National Wildlife Refuge in 1988. Report, Order No. PB92-105923, 43 pp. Avail. NTIS From: Gov. Rep. Announce. Index (U.S.) 1992, 92(4), Abstr. No. 208,515. Palmer Locarnini, S.J. and B.J. Presley. 1995. Trace element concentrations in Antarctic krill, Euphausia superba . Polar Biol. 15(4):283-288. Palmisano, F., N. Cardellicchio and P.G. Zambonin. 1995. Speciation of mercury in dolphin liver: A two-stage mechanism for the demethylation accumulation process and role of selenium. Mar. Environ. Res. 40(2):109-121. Paludan Miller, P., C.T. Agger, R. Dietz and C.C. Kinze. 1993. Mercury, Cadmium, Zinc, Copper And Selenium In Harbour Porpoise Phocoena phocoenaFrom West Greenland. Polar Biol. 13(5):311-320. Papadopoulou, C. and J. Andreotis. 1985. Mercury and selenium concentration in edible fish and plankton from the Aegean Sea. Heavy Met. Environ., Int. Conf., 5th, Volume 1, 733-5. Editor(s): Lekkas, Themistokles D. CEP Consult.: Edinburgh, UK. Paripatananont, T. and R.T. Lovell. 1997. Comparative net absorption of chelated and inorganic trace minerals in channel catfish Ictalurus punctatusdiets. J. World Aquacul. Soc. 28(1):62-67. Park, J. and B.J. Presley. 1997. Trace metal contamination of sediments and organisms from the Swan Lake area of Galveston Bay. Environ. Pollut. 98(2):209-221. Park, K.S., N.B. Kim, Y.S. Kim, K.Y. Lee, S.K. Chun and Y.Y. Yoon. 1994. A Survey of Trace Elements in Fresh-Water Fish and Rice Along the Han River by Neutron Activation Analysis. Biol. Trace Element Res. 43-45(0):229-237.

### Draft November 12, 2004 Ref-66 Patel, B. and J.P. Chandy. 1987. Do selenium and glutathione (GSH) detoxify mercury in marine invertebrates?: II. Effects on gill ATPase and related blood factors in an arcid clam Anadara granosa . Dis. Aquat. Organ. 3(2):127-136. Patel, B., J.P. Chandy, and S. Patel. 1988. Do selenium and glutathione inhibit the toxic effects of mercury in marine lamellibranchs? Sci. Total Environ. 76(2-3):147-166. Patel, B., J.P. Chandy and S. Patel. 1990. Effect of mercury, selenium, and glutathione on sulfhydryl levels and glutathione reductase in blood clam Anadara granosa(L.). Indian J. Mar. Sci. 19(3):187-190. Patrick, R., T. Bott and R. Larson. 1975. The role of trace elements in management of nuisance growths. PB-241985. National Technical Information Service, Springfield, VA. Paulsson, K. and K. Lundbergh. 1991. Treatment of Mercury Contaminated Fish by Selenium Addition. Water Air Soil Pollut. 56:833-841. Paveglio, F.L., C.M. Bunck and G.H. Heinz. 1994. Selenium and boron in aquatic birds from central California. J. Wildl. Manage. 56(1):31-42. Paveglio, F.L., K.M. Kilbride and C.M. Bunck. 1997. Selenium in aquatic birds from central California. J. Wildl. Manage. 61(3):832-839. Payer, H.D. and K.H. Runkel. 1978. Environmental pollutants in freshwater alga from open-air mass cultures. Arch. Hydrobiol. Beih. Ergebn. Limnol. 11:184-198. Payer, H.D., K.H. Rundel, P. Schramel, E. Stengel, A. Bhumiratana and C.J. Soeder. 1976. Environmental influences on the accumulation of lead, cadmium, mercury, antimony, arsenic, selenium, bromine and tin in unicellular algae cultivated in Thialand and in Germany. Chemosphere 6:413-418. Peck, R.K. 1986. The trace element selenium induces trichocyst formation in the ciliated protozoan Pseudomicrothorax dubius . European J. Cell Biol. 41(2):174-181. Pelletier, E. 1986a. Modification of the bioaccumulation of selenium by Mytilus edulisin the presence of organic and inorganic mercury. Can. J. Fish. Aquat. Sci. 43:203-210. Pelletier, E. 1986b. Mercury-selenium interactions in aquatic organisms: A review. Mar. Environ. Res. 18:111-132. Pelletier, E. 1988. Acute toxicity of some methylmercury complexes to Mytilus edulisand lack of selenium protection. Mar. Pollut. Bull. 19(5):213-219. Pennington, C.H., J.A. Baker and M.E. Potter. 1982. Contaminant levels in fishes from Browns Lake, Mississippi. J. Miss. Acad. Sci. 27:139-147. Perez Campo, R., M. Lopez Torres and G. Barja De Quiroga. 1990. Thermal acclimation hydroperoxide detoxifying enzymes and oxidative stress in the lung and liver of Rana perezi . J. Thermal Biol. 15(3-4):193-200.

### Draft November 12, 2004 Ref-67 Perez-Trigo, E., P. Garcia-Martinez, J.L. Catoira and G. Mosquera. 1995. Subcellular distribution of antioxidant enzymes in the gonads of the sea urchin, Paracentrotus lividusLmk, from the Ria Ares-Betanzos, NW Spain. In: Echinoderm Res.: Proc. Eur. Echinoderms Colloq., 4th. 51-55. Emson, R., A. Smith and A. Campbell (Eds). Balkema: Rotterdam, Neth. pp.51-55. Peterson, J.A. and A.V. Nebeker. 1992. Estimation of waterborne selenium concentrations that are toxicity thresholds for wildlife. Arch. Environ. Contam. Toxicol. 23(2):154-162. Petrucci, F., S. Caimi, G. Mura and S. Caroli. 1995. Artemiaas a bioindicator of environmental contamination by trace elements. Microchem. J. 51(1-2):181-186. Phadnis, A.P., B. Nanda, S.A. Patwardhan, P. Powar, and R.N. Sharma. 1988. Products active on mosquitoes: Part III. Synthesis of biologically active 3,7-dimethyl-6-octene-1,8-diol diethers. Indian J. Chem. Section B: Organic Chemistry Including Medicinal Chemistry 27(9):867-870. Phillips, G.R. and R.W. Gregory. 1980. Accumulation of selected elements (As, Cu, Hg, Pb, Se, Zn) by northern pike (

Esox lucius ) reared in surface coal mine decant water. Proc. Mont. Acad. Sci. 39:44-50. Phillips, G.R. and R.C. Russo. 1978. Metal bioaccumulation in fishes and aquatic invertebrates: A literature review. EPA-600/3-78-103. National Technical Information Service, Springfield, VA. Phillips, G.R., P.A. Medvick, D.R. Skaar and D.E. Knight. 1987. Factors affecting the mobilization, transport, and bioavailability of mercury in reservoirs of the Upper Missouri River Basin (USA). U S Fish and Wildlife Service Fish and Wildlife Technical Report 0(10):1-64. Pinto, G. and R.Taddei. 1988. Evaluation of toxic effects of heavy metals on unicellular algae. V - Analysis of the inhibition manifesting itself with an increased lag phase. Boll. Soc. Nat. Napoli, Volume Date 1986, 95:303-17. Poppe, T.T., T. Hastein, A. Froslie, N. Koppang and G. Norheim. 1986. Nutritional aspects of hemorrhagic syndrome ("Hitra disease") in farmed Atlantic salmon Salmo salar . Dis. Aquat. Organ. 1(3):155-162. Poston, H.A., G.G. Combs, Jr. and L. Leibovitz. 1976. Vitamin E and selenium interrelations in the diet of Atlantic salmon (

Salmo salar). Gross histological and biochemical deficiency signs. J. Nutr. 106:892-904. Pratt, J.R. and N.J. Bowers. 1990. Effect of selenium on microbial communities in laboratory microcosms and outdoor streams. Toxicity Assess. 5(3):293-308. Presley, B.J. 1997. Trace metal contamination of sediments and organisms from the Swan Lake area of Galveston Bay. Environ. Pollut. 98(2):209-221. Presley, B.J., R.J. Taylor and P.N. Boothe. 1990. Trace metals in Gulf of Mexico oysters. Sci. Total Environ. 97-98:551-593. Presser, T.S. 1994. "The Kesterson effect". Environ. Manage. 18(3):437-454.

### Draft November 12, 2004 Ref-68 Presser, T.S. and H.M. Ohlendorf. 1987. Biogeochemical cycling of selenium in the San Joaquin Valley, California, USA. Environ. ManagE. 11(6):804-822. Prevot, P. and M.O. Soyer-Gobillard. 1986. Combined action of cadmium and selenium on two marine dinoflagellates in culture,Prorocentrum micans and Crypthecodinium cohnii . J. Protozool. 33(1):42-47. Price, N.M. 1987. Urea and selenium nutrition of marine phytoplankton: a physiological and biochemical study. Avail. NLC From: Diss. Abstr. Int. B 1988, 49(5):1498-9. Price, N.M. and P.J. Harrison. 1988. Specific selenium-containing macromolecules in the marine diatom Thalassiosira pseudonana . Plant Physiol. (Bethesda) 86(1):192-199. Price, N.M., P.A. Thompson and P.J. Harrison. 1987. Selenium: An essential element for growth of the coastal marine diatom Thalassiosira pseudonana (Bacillariophyceae). J. Phycol. 23(1):1-9. Pritchard, T. 1997. Environmental performance of Sydney's deepwater outfalls. Water (Artarmon, Aust.), 24(2):29-34. Pyron, M and T.L. Beitinger. 1989. Effect of selenium on reproductive behavior and fry of fathead minnows. Bull. Environ. Contam. Toxicol. 42(4):609-13. Quevauviller, P., K. Vercoutere, H. Muntau and B. Griepink. 1993a. The certification of the contents (mass fractions) of arsenic, cadmium, chromium, copper, mercury, manganese, nickel, lead, selenium, vanadium and zinc in plankton. Comm. Eur. Communities, [Rep.] EUR, EUR 14558, 71 pp. Quevauviller, P., E.A. Maier and B. Griepink. 1993b. Projects for the improvement of the quality of chemical speciation analyses in environmental matrixes. Fresenius' J. Anal. Chem. 345(2-4):282-286. Rady, A.A., N. Saber, H.M. Kotkat, B. Matkovics and A.M. Nour. 1992. Metals effect on fish tissues I: Effects of chronic mercury and selenium treatment on young tilapia tissue enzymes and lipid peroxidation. Acta Universitatis Szegediensis Acta Biologica 38(1-4):3-9. Ramakrishna, T., K.A. Naidu, S. Vatsala, O. Sreekumar, V.N. Kumar, and K.K. Soudamini. 1988. Selenite neutralizes the toxic effect of cadmium. Indian J. Environ. Health 30(4):355-359. Ramos, F., M.D.C. Castilho, M.I. Noronha da Silveira. 1992. Determination of selenium level in fish. In: Mol. Biol. Atheroscler., [Ed. Proc. Eur. Atheroscler. Soc. Meet.], Meeting Date 1991. Halpern, M.J. (Ed). Libbey: London, UK. pp. 539-540. Rani, P. and K. Lalitha. 1996. Evidence for altered structure and impaired mitochondrial electron transport function in selenium deficiency. Biol. Trace Element Res. 51(3):225-234. Rao, V.R., S.V. Mitz, C.T. Hadden and B.W. Cornaby. 1996. Distribution of contaminants in aquatic organisms from East Fork Poplar Creek. Ecotoxicol. Environ. Saf. 33(1):44-54. Raptis, S.E., G. Kaiser and G. Tolg. 1983. A survey of selenium in the environment and a critical review of its determination at trace levels. Z. Anal. Chem. 316:105-123.

### Draft November 12, 2004 Ref-69 Rauscher, J.D. 1988. Toxic effects of selenium and copper on the planarian, Dugesia dorotocephala . 184 pp. Avail. Univ. Microfilms Int., Order No. DA8827844 From: Diss. Abstr. Int. B 1989, 49(10):4191-2. Reading, J.T. 1979. Acute and chronic effects of selenium on Daphnia pulex . M.S. thesis. Virginia Polytechnic Institute and State University, Blacksburg, VA. Reading, J.T. and A.L. Buikema, Jr. 1980. Effects of sublethal concentrations of selenium on metabolism and filtering rate of Daphnia pulex . Bull. Environ. Contam. Toxicol. 24(6):929-935. Reading, J.T. and A.L. Buikema, Jr. 1983. Chronic effects of selenium on Daphnia pulex . Arch. Environ. Contam. Toxicol. 12:399-404. Reamer, D.C. and W.H. Zoller. 1980. Selenium biomethylation products from soil and sewage sludge. Science 208:500-502. Reash, R.J, J.H. Van Hassel and K.V. Wood. 1988. Ecology of a southern Ohio stream receiving fly ash pond discharge: changes from acid mine drainage conditions. Arch. Environ. Contam. Toxicol. 17(4):543-54. Reash, R.J., T.W. Lohner, K.V. Wood, and V.E. Willet. 1999. Ecotoxicological assessment of bluegill sunfish inhabiting a selenium-enriched fly ash stream. In Environmental Toxicology and Risk Assessment: Standardization of Biomarkers for Endocrine Disruption and Environmental Assessment: Eighth Volume, ASTM STP 1364 (D.S. Henshel, M.C. Black and M.C. Harris Eds. Regoli, F. 1998. Trace metals and antioxidant enzymes in gills and digestive gland of Mediterranean mussel Mytilus galloprovincialis . Arch. Environ. Contam. Toxicol. 34(1):48-63. Regoli, F. and G. Principato. 1995. Glutathione, glutathione-dependent and antioxidant enzymes in mussel, Mytilus galloprovincialis , exposed to metals under field and laboratory conditions: Implications for the use of biochemical biomarkers. Aquat. Toxicol. (Amsterdam) 31(2):143-164. Regoli, F., G.B. Principato, E. Bertoli, M. Nigro, and E. Orlando. 1997. Biochemical characterization of the antioxidant system in the scallop Adamussium colbecki, a sentinel organism for monitoring the Antarctic environment. Polar Biology 17(3):251-258. Reinfelder, J.R. and N.S. Fisher. 1991. The Assimilation of Elements Ingested by Marine Copepods. Science (Washington DC) 251(4995):794-796. Reinfelder, J.R. and N.S. Fisher. 1994a. Retention of elements absorbed by juvenile fish ( Menidia Menidia , Menidia beryllina) from zooplankton prey. Limnol. Oceanogr. 39(8):1783-1789. Reinfelder, J.R. and N.S. Fisher. 1994b. The assimilation of elements ingested by marine planktonic bivalve larvae. Limnol. Oceanogr. 39(1):12-20. Reinfelder, J.R., N.S. Fisher, S.W. Fowler and J.L. Teyssie. 1993. Release rates of trace elements and protein from decomposing planktonic debris 2. Copepod carcasses and sediment trap particulate matter. J. Mar. Res. 51(2):423-442.

### Draft November 12, 2004 Ref-70 Reinfelder, J.R., W.X. Wang, S.N. Luoma, and N.S. Fisher. 1997. Assimilation efficiencies and turnover rates of trace elements in marine bivalves: A comparison of oysters, clams and mussels. Mar. Biol.(Berlin) 129(3):443-452. Renzoni A., S. Focardi, C. Fossi, C. Leonzio and J. Mayol. 1986. Comparison between concentrations of mercury and other contaminants in eggs and tissues of Cory's shearwater Calonectris diomedeacollected on Atlantic and Mediterranean islands. Environ. Pollut. Series A Ecol. Biol. 40(1):17-36. Rhodes, L. and B. Burke. 1996. Morphology and growth characteristics of Chrysochromulinaspecies (Haptophyceae equals Prymnesiophyceae) isolated from New Zealand coastal waters. New Zealand J. Mar. Freshwater Res. 30(1):91-103. Rhodes, L.L., C.J. O'Kelly and J.A. Hall. 1994. Comparison of growth characteristics of New Zealand isolates of the prymnesiophytesChrysochromulina quadrikontaand C. camellawith those of the ichthyotoxic species C. polylepis . J. Plankton Res. 16(1):69-82. Ribeyre, F., C. Amiard Triquet, A. Boudou, and J.C. Amiard. 1995. Experimental study of interactions between five trace elements-Cu, Ag, Se, Zn, and Hg-toward their bioaccumulation by fish ( Brachydanio rerio ) from the direct route. Ecotoxicol. Environ. Safety. 32(1):1-11. Rice, C.A., P.D. Plesha, E. Casillas, D.A. Misitano and J.P. Meador. 1995. Growth and survival of three marine invertebrate species in sediments from the Hudson-Raritan estuary, New York. Environ. Toxicol. Chem. 14(11):1931-1940. Richter, J.E. 1982. Center for Lake Superior Environmental Studies, University of Wisconsin-Superior, Superior, WI. (Memorandum to C.E. Stephan, U.S. EPA, Duluth, MN. June 30.) Richter, D., and H. Bergmann 1993. Selenium Uptake by Wheat Plants. In: M. Anke (ed.),

MengenSpurenelem., 13th Arbeitstag, 1993 . Verlag MTV Hammerschmidt, Gersdorf, Germany. Riedel, G.F. and J.G. Sanders. 1996. The influence of pH and media composition on the uptake of inorganic selenium by Chlamydomonas reinhardtii . Environ. Toxicol. Chem. 15(9):1577-1583. Riedel, G.F., D.P. Ferrier and J.G. Sanders. 1991. Uptake of selenium by freshwater phytoplankton. Water Air Soil Pollut. 57-58:23-30. Riget, F., P. Johansen and G. Asmund. 1996. Influence of length on element concentrations in blue mussels (Mytilus edulis ). Mar. Pollut. Bull. 32(10):745-751. Riggs, M.R. and G.W. Esch. 1987. The suprapopulation dynamics of Bothriocephalus acheilognathiin a North Carolina (USA) reservoir: Abundance, dispersion, and prevalence. J. Parasitol. 73(5):877-892. Riggs, M.R., A.D. Lemly and G.W. Esch. 1987. The growth, biomass, and fecundity of Bothriocephalus acheilognathiin a North Carolina (USA) cooling reservoir. J. Parasitol. 73(5):893-900. Ringdal, D. and K. Julshamn. 1985. Effect of selenite on the uptake of methylmercury in cod ( Gadus morhua ). Bull. Environ. Contam. Toxicol. 35:335-344.

### Draft November 12, 2004 Ref-71 Risenhoover, K.L. 1989. Composition and quality of moose winter diets in interior Alaska (USA). J. Wildl. Manage. 53(3):568-577. Robberecht, H. and R. Van Grieken. 1982. Selenium in environmental waters: Determination, speciation and concentration levels. Talanta 29:823-844. Robertson, A., B.W. Gottholm, D.D. Turgeon and D.A. Wolfe. 1991. A Comparative Study of Contaminant Levels in Long Island Sound USA. Estuaries 14(3):290-298. Roederer, G. 1986. On the toxic effects of tetraethyllead and its derivatives on the chrysophyte Poterioochromonas malhamensis . VII. Protective action of thiol compounds, vitamins, trace elements and other agents. Ecotoxicol. Environ. Saf. 11(3):277-94. Ronald, K., R. J. Frank, J. Dougan, R. Frank and H. E. Braun. 1984. Pollutants in harp seals (Phoca groenlandica): 2. Heavy metals and selenium. Sci. Total Environ. 38(0):153-166. Roper, J.M., D.S. Cherry, J.W. S immers and H.E. Tatem. 1997. Bioaccumulation of toxicants in the zebra mussel, Dreissena polymorpha , at the Times Beach Confined Disposal Facility, Buffalo, New York. Environ. Pollut., Volume Date 1996, 94(2):117-129. Rose, C.D., T.J. Ward and V.E. De Pass. 1985. Ecological assessment for coal ash dumped at deepwater dumpsite-106. Wastes Ocean 4:389-422. Ed(s): Duedall, Iver W. Wiley: New York, N. Y. Rosetta, T.N. and A.W. Knight. 1995. Bioaccumulation of selenate, selenite, and seleno-DL-methionine by the brine fly larvae Ephydra cinereaJones. Arch. Environ. Contam. Toxicol. 29(3):351-357. Rossell, I.M., D.J. Raynal and D.J. Leopold. 1994. The effects of watershed liming on the tissue chemistry of three co-occurring poor fen species. Can. J. Bot. 72(12):1825-1834. Rouleau, C., E. Pelletier and J. Pellerin Massicotte. 1992. Uptake of Organic Mercury and Selenium from Food by Nordic Shrimp Pandalus borealis . Chem. Spec. Bioavail. 4(2):75-81. Roux, D.J., J.E. Badenhorst, H.H. DuPrez and G.J. Steyn. 1994. Note on the occurrence of selected trace metals and organic compounds in water, sediment and biota of the Crocodile River, Eastern Transvaal, South Africa. Water S.A. (Pretoria) 20(4):333-340. Roux, D.J., S.H.J. Jooste and H.M. Mackay. 1996. Substance-specific water quality criteria for the protection of South African freshwater ecosystems: Methods for derivation and initial results for some inorganic toxic substances. S.A. J. Sci. 92(4):198-206. Rudd, J.W.M. and M.A. Turner. 1983a. The English-Wabigoon River system: II. Suppression of mercury and selenium bioaccumulation by suspended and bottom sediments. Can. J. Fish. Aquat. Sci. 40:2218- 2227. Rudd, J.W.M. and M.A. Turner. 1983b. The English-Wabigoon River system: V. Mercury and selenium bioaccumulation as a function of aquatic primary productivity. Can. J. Fish. Aquat. Sci. 40:2251-2259.

### Draft November 12, 2004 Ref-72 Rudd, J.W.M., M.A. Turner, B.E. Townsend, A. Swick and A. Furutani. 1980. Dynamics of selenium in mercury-contaminated experimental freshwater ecosystems. Can. J. Fish. Aquat. Sci. 37:848-857. Ruelle, R., K.D. Keenlyne. 1993. Contaminants in Missouri River pallid sturgeon. Bull. Environ. Contam. Toxicol. 50(6):898-906. Ryther, J., T.M. Losordo, A.K. Furr, T.F. Parkinson, W.H. Gutenman, I.S. Pakkala and D.J. Lisk. 1979. Concentration of elements in marine organisms cultured in seawater flowing through coal-fly ash. Bull. Environ. Contam. Toxicol. 23:207-210. Sager, D.R. and C.R. Cofield. 1984. Differential accumulation of selenium among axial muscle, reproductive and liver tissues of four warmwater fish species. Water Resour. Bull. 20:359-363. Saiki, M.K. 1986a. A field example of selenium contamination in an aquatic food chain. In: Proceedings of the first annual environmental symposium: Selenium in the environment. Calif. Agri. Tech. Inst., Fresno, CA. pp. 67-76. Saiki, M.K. 1986b. Concentrations of selenium in aquatic food-chain organisms and fish exposed to agricultural tile drainage water. In: Selenium and agriculture drainage: Implications for San Francisco Bay and the California environment. The Bay Institute of San Francisco, Tiburon, CA. pp. 25-33. Saiki, M.K. 1987. Relation of length and sex to selenium concentrations in mosquitofish. Environ. Pollut. 47(3): 171-186. Saiki, M.K. 1990. Elemental concentrations in fishes from the Salton Sea, southeastern California. Water, Air, Soil Pollut. 52(1-2):41-56. Saiki, M.K. and M.R. Jenings. 1992. Toxicity of Agricultural Subsurface Drainwater from the San Joaquin Valley California to Juvenile Chinook Salmon and Striped Bass. Trans. Am. Fish. Soc. 121(1):78-93. Saiki, M.K. and T.P. Lowe. 1987. Selenium in aquatic organisms from subsurface agricultural drainage water, San Joaquin Valley, California (USA). Arch. Environ. Contam. Toxicol. 16(6):657-670. Saiki, M. and T.W. May. 1988. Trace element residues in bluegills and common carp from the lower San Joaquin River, California (USA) and its tributaries. Sci. Total Environ. 74(0):199-218. Saiki, M.K. and R.S. Ogle. 1995. Evidence of impaired reproduction by western mosquitofish inhabiting seleniferous agricultural drainwater. Trans. Am. Fish. Soc. 124(4):578-587. Saiki, M.K. and D.U. Palawski. 1990. Selenium and Other Elements in Juvenile Striped Bass from the San Joaquin Valley and San Francisco Estuary California USA. Arch. Environ. Contam. Toxicol. 19(5):717-730. Saiki M.K., M.R. Jennings and T.W. May. 1992. Selenium and Other Elements in Freshwater Fishes from the Irrigated San Joaquin Valley California. Sci. Total Environ. 126(1-2):109-137.

### Draft November 12, 2004 Ref-73 Saiki, M.K., M.R. Jennings and W.G. Brumbaugh. 1993. Boron, Molybdenum and Selenium in Aquatic Food Chains from the Lower San Joaquin River and its Tributaries California. Arch. Environ. Contam. Toxicol. 24(3):307-319. Saleh, M.A.S., A. Mostafa, M.M. Fouda, M.A. Saleh, M.S. Abdel Lattif and B.L. Wilson. 1988. Inorganic pollution of the man-made lakes of Wadi El-Raiyan and its impact on aquaculture and wildlife of the surrounding Egyptian desert. Arch. Environ. Contam. Toxicol. 17(3):391-403.Salki, A., M. Turner, K. Patalas, J. Rudd and D. Findlay. 1985. The influence of fish-zooplanktonphytoplankton interactions on the results of selenium toxicity experiments within large enclosures. Can. J. Fish. Aquat. Sci. 42:1132-1143. Salte, R., T. Asgard, and K. Liestol. 1988. Vitamin E and selenium prophylaxis against "Hitra disease" in farmed Atlantic Salmon: A survival study. Aquaculture 75(1-2):45-56. Sanders, R.W. and C.C. Gilmour. 1994. Accumulation of selenium in a model freshwater microbial food web. Appl. Environ. Microbiol. 60(8):2677-2683. Sandholm, M., H.E. Oksanen and L. Pesonen. 1973. Uptake of selenium by aquatic organisms. Limnol. Oceanogr. 18:496-499. Sarathchandra, S.U. and J.H. Watkinson. 1981. Oxidation of elemental selenium to selenite by Bacillus megaterium . Science 211:600-601. Sarma, Y.S.R.K. and S. Jayaraman. 1984. Observations on sulphur-selenium antagonism on the growth of two desmids. Acta Bot. Indica 12:57-60. Sastry, K.V. and V. Shukla. 1994. Influence of protective agents in the toxicity of cadmium to a freshwater fish ( Channa punctatus ). Bull. Environ. Contam. Toxicol. 53(5):711-717. Sato, T., Y. Ose and T. Sakai. 1980. Toxicological effect of selenium on fish. Environ. Pollut. 21A:217- 224. Savant, K.B. and G.V. Nilkanth. 1991. On comparative studies of acute toxicity of hexavalent chromium and selenium to Scylla seratta(Forskal). Pollut. Res. 10(4):239-243. Scanes, P. 1997. "Oyster watch": monitoring trace metal and organochlorine concentrations in Sydney's coastal waters. Mar. Pollut. Bull., Volume Date 1996, 33(7-12):226-238. Schantz, M.M., R. Demiralp, R.R. Greenberg, M.J. Hays, R.M. Parris, B.J. Porter, D.L. Poster, L.C. Sander, K.S. Sharpless, S.A. Wise, and S.B. Schiller. 1997. Certification of a frozen mussel tissue standard reference material (SRM 1974a) for trace organic constituents. Fresenius' J. Anal. Chem., 358(3):431-440. Scheuhammer, A.M., A.H.K. Wong, and D. Bond. 1998. Mercury and selenium accumulation in common loons ( Gavia immer

) and common mergansers (Mergus merganser ) from eastern Canada. Environ. Toxicol. Chem. 17(2):197-201.

### Draft November 12, 2004 Ref-74 Schmitt, C.J. and W.G. Brumbaugh. 1990. National Contaminant Biomonitoring Program Concentrations of Arsenic, Cadmium, Copper, Lead, Mercury, Selenium and Zinc in USA Freshwater Fish 1976-1984. Arch. Environ. Contam. Toxicol. 19(5):731-747. Schmitt, C.J., M.L. Wildhaber, J.B. Hunn, T. Nash, M.N. Tieger and B.L. Steadman. 1993. Biomonitoring of lead-contaminated Missouri streams with an assay for erythrocyte .delta.-aminolevulinic acid dehydratase activity in fish blood. Arch. Environ. Contam. Toxicol. 25(4):464-475. Schramel, P. and L. Xu. 1991. Determination of arsenic, antimony, bismuth, selenium and tin in biological and environmental samples by continuous flow hydride generation inductively coupled plasma-atomic emission spectrometry (ICP-AES) without gas-liquid separator. Fresenius. J. Anal. Chem. 340(1):41-47. Schuler, C.A., R.G. Anthony and H.M. Ohlendorf. 1990. Selenium in wetlands and waterfowl foods at Kesterson Reservoir, California, 1984. Arch. Environ. Contam. Toxicol. 19(6):845-853. 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. Schultz, T.W., S.R. Freeman and J.N. Dumont. 1980. Uptake, depuration, and distribution of selenium in Daphnia and its effects on survival and ultrastructure. Arch. Environ. Contam. Toxicol. 9:23-40. Scott, K.C. and J.D. Latshaw. 1993. Macro and micro mineral levels in the tissues of menhaden fish. J. Aquat. Food Prod. Technol. 2(2):51-61. Secor, C.L., E.L. Mills, J. Harshbarger, H.T. Kuntz, W.H. Gutenmann and D.J. Lisk. 1993. Bioaccumulation of Toxicants, Element and Nutrient Composition and Soft Tissue Histology of Zebra Mussels Dreissena polymorphafrom New York State Waters. Chemosphere 26(8):1559-1575. Seelye, J.G., R.J. Hesselberg and M.J. Mac. 1982. Accumulation by fish of contaminants released from dredged sediments. Environ. Sci. Technol. 16:459-464. Segner, H., D. Lenz, W. Hanke and G. Schueuermann. 1994. Cytotoxicity of metals toward rainbow trout R1 cell line. Environ. Toxicol. Water Qual. 9(4):273-279. Sen, S., S. Mondal, J. Adhikari, D. Sarkar, S. Bose, B. Mukhopadhyay and S. Bhattacharya. 1995. Inhibition of fish brain acetylcholinesterase by cadmium and mercury. Interaction with selenium. In: Enzymes Cholinesterase Fam., [Proc. Int. Meet. Cholinesterases], 5th, Meeting Date 1994. Quinn, D.M. (Ed). Plenum: New York, N.Y. pp.369-374. Sevareid, R. And G. Ichikawa. 1983. Physiological stress (scope for growth) of mussels in San Francisco bay. Waste Disposal Oceans: Minimizing Impact, Maximizing Benefits, [Pap. - South. Calif. Acad. Sci. Symp., "Ocean Disposal 1980s"], Meeting Date 1982, 152-70. Ed(s): Soule, Dorothy F.; Walsh, Don. Westview: Boulder, Colo. (English) 1983. Shabana, E.F. and S.A. El-Attar. 1995. Influence of clay minerals on selenium toxicity to algae. Egypt. J. Microbiol., Volume Date 1995, 30(2):275-286.

### Draft November 12, 2004 Ref-75 Shamberger, R.J. 1983. Biochemistry of selenium. Plenum Press, New York, NY. Sharif, A.K.M., M. Alamgir, K.R. Krishnamoorthy and A.I. Mustafa. 1993. Determination of arsenic, chromium, mercury, selenium and zinc in tropical marine fish by neutron activation. J. Radioanal. Nucl. Chem. 170(2):299-307. Sharma, D.C. and P.S. Davis. 1980. Behavior of some radioactive compounds of mercury and selenium in aquarium water and their direct uptake by the goldfish Carassius auratus . Ind. J. Exp. Biol. 18:69-71. Sheline, J. and B. Schmidt-Nielsen. 1977. Methylmercury-selenium: Interaction in the killfish, Fundulus heteroclitus . In: Physiological responses of marine biota to pollutants. Vernberg, F.J., A. Calabrese, F.P. Thurberg, and W.B. Vernberg (Eds.). Academic Press, New York, NY. pp. 119-130. Shen, L.H., M.H.V. Nieuwenhuizen, and J.B. Luten. 1997. Speciation and in vitro bioavailability of selenium in fishery products. Dev. Food Sci. 38: 653-663. Shigeoka, S., T. Takeda, T. Hanaoka, A. Yokota, S. Kitaoka and Y. Iizuka. 1990. Properties of selenium-induced glutathione peroxidase in low-carbon dioxide-grown Chlamydomonas reinhardtii . Curr. Res. Photosynth., Proc. Int. Conf. Photosynth., 8th, Meeting Date 1989, Volume 4. Baltscheffsky, M. (Ed). Kluwer: Dordrecht, Neth. pp. 615-618. Shigeoka, S., T. Takeda and T. Hanaoka. 1991. Characterization and immunological properties of selenium-containing glutathione peroxidase induced by selenite in Chlamydomonas reinhardtii . Biochem. J. 275(3):623-628. Shinohara, K., Y. Okura, T. Koyano, H. Murakami, E. H. Kim and H. Omura. 1986. Growth-promoting effects of an extract of a thermophilic blue-green alga, Synechococcus elongatus var. on human cell lines. Agric. Biol. Chem. 50(9):2225-2230. Shirasaki, T., J. Yoshinaga, M. Morita, T. Okumoto and K. Oishi. 1996. An application of nitrogen microwave-induced plasma mass spectrometry to isotope dilution analysis of selenium in marine organisms. Tohoku J Exp. Med. 178(1):81-90. Short, T.M. and C.G. Wilber. 1980. Effects of chronic exposure to sodium selenite on calcium balance in the crayfish Orconectes immunis . Am. Zool. 20:801. Shrift, A. 1954a. Sulfur-selenium antagonism. I. Antimetabolic action of selenate on the growth of Chlorella vulgaris . Am. J. Bot. 41:223-230. Shrift, A. 1954b. Sulfur-selenium antagonism. II. Antimetabolic action of seleno-methionine on the growth of Chlorella vulgaris . Am. J. Bot. 41:345-352. Shrift, A. 1961. Biochemical interrelations between selenium and sulfur in plants and microorganisms. Fed. Proc. 20:695-702. Shrift, A. 1964. A selenium cycle in nature. Nature 201:1304-1305.

### Draft November 12, 2004 Ref-76 Shrift, A. 1973. Metabolism of selenium by plants and microorganisms. In: Organic selenium compounds: Their chemistry and biology. Klayman, D.L. and W.H.H. Gunther (Eds.). Wiley-Interscience, New York, NY. pp. 763-814. Shultz, C.D. and B.M. Ito. 1979. Mercury and selenium in blue marlin, Mahaira nigricans , from Hawaiian Islands. Fish. Bull. 76:872-879. Siebers, D. and U. Ehlers. 1979. Heavy metal action in transintegumentary absorption of glycine in two annelid species. Mar. Biol. (Berl.) 50:175-179. Siegel, B.Z., S.M. Siegel, T. Correa, C. Dagan, G. Galvez, L. Leeloy, A. Padua and E. Yaeger. 1991. The Protection of Invertebrates Fish and Vascular Plants Against Inorganic Mercury Poisoning by Sulfur and Selenium Derivatives. Arch. Environ. Contam. Toxicol. 20(2):241-246. Simopoulos, A.P. 1997. Nutritional aspects of fish. Dev. Food Sci. 38: 589-607. Siwicki, A.K., D.P. Anderson and G.L. Rumsey. 1994. Dietary intake of immunostimulants by rainbow trout affects non-specific immunity and protection against furunculosis. Vet. Immunol. Immunopathol. 41(1-2):125-139. Skaare, J.U., N.H. Markussen, G. Norheim, S. Haugen and G. Holt. 1990. Levels of Polychlorinated Biphenyls, Organochlorine Pesticides, Mercury, Cadmium, Copper, Selenium, Arsenic and Zinc in the Harbor Seal Phoca vitulinain Norwegian Waters. Environ. Pollut. 66(4):309-324. Skaare, J.U., E. Degre, P.E. Aspholm and K.I. Ugland. 1994. Mercury and selenium in Arctic and coastal seals off the coast of Norway. Environ. Pollut. 85(2):153-160. Skerfving, S. 1978. Interaction between selenium and methylmercury. Environ. Health Perspect. 25:57-65. Skinner, W. F. 1985. Trace element concentrations in wastewater treatment basin-reared fishes: results of a pilot study. Proc. Pa. Acad. Sci. 59(2):155-61. Skorupa, J.P., S.P. Morman and J.S. Sefchick. 1996. Guidelines for interprreting selenium exposures of biota associated with non-marine aquatic habitats. U.S. Department of Interior, Fish and Wildlife Service, Sacramento, Field Office, Sacromento, CA., 74 pp. Smith, D.R. and A.R. Flegal. 1989. Elemental concentrations of hydrothermal vent organisms from the Galapagos Rift (Ecuador). Mar. Biol. (Berlin) 102(1):127-134. Smith, I.R., A.F. Johnson, D. MacLennan, H. Manson. 1992. Chemical contaminants, lymphocystis, and dermal sarcoma in walleyes spawning in the Thames River, Ontario. Trans. Am. Fish. Soc. 121(5):608-616. Smith, L.L., Jr., D.M. Oseid, G.L. Kimball and S.M. El-Kandelgy. 1976. Toxicity of hydrogen sulfide to various life history stages of bluegill ( Lepomis macrochirus). Trans. Am. Fish. Soc. 105:442-449.

### Draft November 12, 2004 Ref-77 Snell, T.W., B.D. Moffat, C. Janssen and G. Persoone. 1991a. Acute toxicity tests using rotifers. III. Effects of temperature, strain, and exposure time on the sensitivity of Brachionus plicatilis . Environ. Toxicol. Water Qual. 6(1):63-75. Snell, T.W., B.D. Moffat, C. Janssen and G. Persoone. 1991b. Acute toxicity tests using rotifers. IV. Effects of cyst age, temperature, and salinity on the sensitivity of Brachionus calyciflorus . Ecotoxicol. Environ. Safety. 21(3):308-317. Sokal, R.R. and F.J. Rohlf. 1981. Biometry. W.H. Freeman and Company, New York, NY. Somerville, H.J., D. Bennett, J.N. Davenport, M.S. Holt, A. Lynes, A. Mahieu, B. McCourt, J.G. Parker and R.R. Stephenson. 1987. Environmental effect of produced water from North Sea oil operations. Mar. Pollut. Bull. 18(10):549-58. Sorensen, E.M.B. 1988. Selenium accumulation, reproductive status, and histopathological changes in environmentally exposed redear sunfish. Arch. Toxicol. 61(4):324-329. Sorensen, E.M.B. and T.L. Bauer. 1983. Hematological dyscrasia in teleosts chronically exposed to selenium-laden effluent. Arch. Environ. Contam. Toxicol. 12:135-141. Sorensen, E.M.B. and T.L. Bauer. 1984a. Planimetric analysis of redear sunfish ( Lepomis microlophus ) hepatopancreas following selenium exposure. Environ. Toxicol. Chem. 3:159-165. Sorensen, E.M.B. and T.L. Bauer. 1984b. A correlation between selenium accumulation in sunfish and changes in condition factor and organ weight. Environ. Pollut. 34A:357-366. Sorensen, E.M.B. and P. Bjerregaard. 1991. Interactive accumulation of mercury and selenium in the sea star Asterias rubens . Mar. Biol. (Berlin) 108(2):269-276. Sorensen, E.M.B., T.L. Bauer, J.S. Bell and C.W. Harlan. 1982. Selenium accumulation and cytotoxicity in teleosts following chronic, environmental exposure. Bull. Environ. Contam. Toxicol. 29:688-696. Sorensen, E.M.B., C.W. Harlan, J.S. Bell, T.L. Bauer and A.H. Prodzynski. 1983. Hepatocyte changes following selenium accumulation in a freshwater teleost. Am. J. Forensic. Med. Pathol. 4:25-32. Sorensen, E.M.B., P.M. Cumbie, T.L. Bauer, J.S. Bell and C.W. Harlan. 1984. Histopathological, hemotological, condition-factor, and organ weight changes associated with selenium accumulation in fish from Belews Lake, North Carolina. Arch. Environ. Contam. Toxicol. 13:153-162. Southworth, G. R., M.J. Peterson and R.R. Turner. 1994. Changes in concentrations of selenium and mercury in largemouth bass following elimination of fly ash discharge to a quarry. Chemosphere 29(1):71-79. Sparling, D.W. and T.P. Lowe. 1996. Metal concentrations of tadpoles in experimental ponds. Environ. Pollut. 91(2):149-159.

### Draft November 12, 2004 Ref-78 Specht, W.L., D.S. Cherry, R.A. Lechleitner and J. Cairns, Jr. 1984. Structural, functional, and recovery responses of stream invertebrates to fly ash effluent. Can. J. Fish. Aquat. Sci. 41:884-896. Spehar, R.L. 1986. U.S. EPA, Duluth, MN. (Memorandum to D.J. Call, Center for Lake Superior Environmental Studies, University of Wisconsin-Superior, Superior, WI. September 16.) Speyer, M.R. 1980. Mercury and selenium concentrations in fish, sediments, and water of two northwestern Quebec lakes. Bull. Environ. Contam. Toxicol. 24:427-432. Srivastava, A.K. and A.K. Srivastava. 1995. Histopathological changes in the liver associated with selenium exposure in the freshwater Indian catfish Heteropneustes fossilis . J. Adv. Zool. 16(1):30-33. Srivastava, D.K. and R.K. Tyagi. 1985. Toxicity of selenium and vanadium to the striped gourami, Colisa fasciatus . Acta Hydrobiol. 25-26(3-4):481-486. Stadtman, T.C. 1974. Selenium biochemistry. Science 183:915-922. Stanley, T.R. Jr, J.W. Spann, G.J. Smith and R. Rosscoe. 1994. Main and interactive effects of arsenic and selenium on mallard reproduction and duckling growth and survival. Arch. Environ. Contam. Toxicol. 26(4):444-451. Stanley, T.R. Jr., G.J. Smith, D.J. Hoffman, G.H. Heinz and R. Rosscoe. 1996. Effects of boron and selenium on mallard reproduction and duckling growth and survival. Environ. Toxicol. Chem. 15(7):1124-1132. Steele, C.W., S. Strickler Shaw and D.H. Taylor. 1992. Attraction of Crayfishes Procambarus clarkii, Orconectes rusticusand Cambarus bartoni to a Feeding Stimulant and its Suppression by a Blend of Metals. Environ. Toxicol. Chem. 11(9):1323-1329. Steimle, F.W., V.S. Zdanowicz, S.L. Cunneff and R. Terranova. 1994. Trace metal concentrations in common benthic macrofaunal prey from the New York Bight apex. Mar. Pollut. Bull. 28(12):760-765. Stemmer, B.L., G.A. Burton Jr. and S. Leibfritz-Frederick. 1990. Effect of sediment test variables on selenium toxicity toDaphnia magna . Environ. Toxicol. Chem. 9(3):381-390. Stephan, C.E., D.I. Mount, D.J. Hansen, J.H. Gentile, G.A. Chapman and W.A. Brungs. 1985. Guidelines for deriving numerical national water quality criteria for the protection of aquatic organisms and their uses. PB85-227049. National Technical Information Service, Springfield, VA. Stoeppler, M., F. Backhaus, M. Burow, K. May and C. Mohl. 1988. Comparative investigations on trace metal levels in brown algae and common (blue) mussels at the same location in the Baltic Sea and the North Sea. NBS Spec. Publ. (U. S.), 740 (Prog. Environ. Specimen Banking), 53-6. Stone, S.T., D.A. Becker, B.J. Koster, P.A. Pella, G. Sleater, M. P.M. Tillekeratne, R. Zeisler and R.W. Sanders. 1988. Inorganic analytic methods and results for marine bivalves and sediments. NBS Spec. Publ. (U.S.), 740 (Prog. Environ. Specimen Banking):62-73.

### Draft November 12, 2004 Ref-79 Stripp, R.A., M. Heit, D.C. Bogen, J. Bidanset and L. Trombetta. 1990. Trace element accumulation in the tissues of fish from lakes with different pH values. Water Air Soil Pollut. 51(1-2):75-87. Stumm, W. and J.J. Morgan. 1982. Aquatic chemistry. Wiley, New York, NY. pp.176-177. Summers, J.K., J.F. Paul and A. Robertson. 1995. Monitoring the ecological condition of estuaries in the United States. Toxicol. Environ. Chem. 49(1+2): 93-108. Sun, L., C.S. Bradford, C. Ghosh, P. Collodi and D.W. Barnes. 1995. ES-like cell cultures derived from early zebrafish embryos. Molec. Mar. Biol. Biotechnol. 4(3):193-199. Sundarrao, K., J. Tinkerame, C. Kaluwin, K. Singh and T. Matsuoka. 1991. Lipid content, fatty acid, and mineral composition of mud crabs ( Scylla serrata ) from Papua New Guinea. J. Food Compos. Anal. 4(3):276-280. Sundarrao, K., J. Tinkerame, C. Kaluwin, K. Singh and T. Matsuoka. 1992. Fatty Acid and Mineral Composition of ShellfishGeloina papua . Fish, Technol. 29(2):144-146. Svensson, B.G., A. Schutz, A. Nilsson, I. Aakesson, B. Aakesson and S. Skerfving. 1992. Fish as a source of exposure to mercury and selenium. Sci. Total Environ. 126(1-2):61-74. Szilagyi, M., J. Nemcsok, S. Sankari, A. Suri and E. Szabo. 1993. Effects of selenium supplementation on serum biochemical parameters in paraquat poisoned carp. Mengen Spurenelem. Arbeitstag. 13th. Anke, M. (Ed). Verlag MTV Hammerschmidt: Gersdorf, Germany. pp. 155-162. Tabaka, C.S., D.E. Ullrey, J.G. Sikarskie, S.R. Debar, and P.K. Ku. 1996. Diet, cast composition, and energy and nutrient intake of red-tailed hawks ( Buteo jamaicensis), great horned owls ( Bubo virginianus ), and turkey vultures (

Cathartes aura ). J. Zoo Wildl. Med. 27(2):187-196. Tafro, A and M. Kiskaroly. 1986. The importance of some vitamins in the nutrition of cyprinid fish. Veterinarski Glasnik 40(6):463-469. Takayangi, K. and D. Cossa. 1985. Speciation of dissolved selenium in the upper St. Lawrence estuary. In: Marine and estuarine geochemistry. Sigleo, A.C. and A. Hattori (Eds.). Lewis Publishers, Chelsea, MI. pp. 275-284. Takayangi, K. and G.T.F. Wong. 1984. Total selenium and selenium(IV) in the James River estuary and southern Chesapeake Bay. Estuarine Coastal Shelf Sci. 18:113-119. Takeda, T., S. Shigeoka, O. Hirayama and T. Mitsunaga. 1992a. The presence of enzymes related to glutathione metabolism and oxygen metabolism in Chlamydomonas reinhardtii . Biosci. Biotechnol. Biochem. 56(10):1662-1663. Takeda, T., S. Shigeoka and T. Mitsunaga. 1992b. Induction of glutathione peroxidase by selenite and its physiological function inChlamydomonas reinhardtii . Phosphorus, Sulfur Silicon Relat. Elem. 67(1-4):439-444.

### Draft November 12, 2004 Ref-80 Takeda, T., Y. Nakano and S. Shigeoka. 1993. Effects of selenite, CO-2 and illumination on the induction of selenium-dependent glutathione peroxidase in Chlamydomonas reinhardtii . Plant Sci. (Limerick) 94(1-2):81-88. Takeda, T., T. Ishikawa and S. Shigeoka. 1997. Metabolism of hydrogen peroxide by the scavenging system in Chlamydomonas reinhardtii . Physiol. Plant. 99(1):49-55. Talbot, V. and W.J. Chang. 1987. Rapid multielement analysis of oyster and cockle tissue using x-ray fluorescence spectrometry, with application to reconnaissance marine pollution investigations. Sci. Total Environ. 66:213-23. Tallandini, L., R. Cecchi, S. De Boni, S. Galassini, G. Ghermandi, G. Gialanella, N. Liu, R. Moro, M. Turchetto and Y. Zhang. 1996. Toxic levels of selenium in enzymes and selenium uptake in tissues of a marine fish. Biol. Trace Element Res. 51(1):97-106. Tan, Y. and W.D. Marshall. 1997. Enzymic digestion-high-pressure homogenization prior to slurry introduction electrothermal atomic absorption spectrometry for the determination of selenium in plant and animal tissues. Analyst (Cambridge, U. K.) 122(1):13-18. Tanizaki, Y., T. Shimokawa, and M. Nakamura. 1992. Physicochemical Speciation of Trace Elements in River Waters by Size Fractionation, ” Environmental Science of Technology . 26:1433-1443. Tang, S.M., I. Orlic, K.N. Yu, J.L. Sanchez, P.S.P. Thong, F. Watt, and H.W. Khoo. 1997. Nuclear microscopy study of fish scales. Nucl. Instrum. Meth. Phys. Res., Sect. B, 130(1-4):396-401. Tao, H., J.W.H. Lam and J.W. McLaren. 1993. Determination of selenium in marine certified reference materials by hydride generation inductively coupled plasma mass spectrometry. J. Anal. At. Spectrom. 8(8):1067-1073. Tao, J., P. Kellar, and W. Warren-Hicks. 1999. Statistical analysis of selenium toxicity data. Report submitted for US EPA, Health and Ecological Criteria Division. The Cadmus Group, Inc., Durnham, NC. Teherani, D.K. 1987. Trace elements analysis in rice. J. Radioanal. Nucl. Chem. 117(3):133-144. Teigen, S.W., J.U. Skaare, A. Bjorge, E. Degre and G. Sand. 1993. Mercury And Selenium In Harbor Porpoise Phocoena phocoenaIn Norwegian Waters. Environ. Toxicol. Chem. 12(7):1251-1259. Thomas, W.H., J.T. Hollibaugh and D.L.R. Siebert. 1980a. Effects of heavy metals on the morphology of some marine phytoplankton. Phycologia 19:202-209. Thomas, W.H., J.T. Hollibaugh, D.L.R. Siebert and G.T. Wallace, Jr. 1980b. Toxicity of a mixture of ten metals to phytoplankton. Mar. Ecol. Prog. Ser. 2:213-220. Thompson, P.A. and W. Hosja. 1996. Nutrient limitation of phytoplankton in the Upper Swan River Estuary, Western Australia. Mar. Freshwater Res. 47(4):659-667.

### Draft November 12, 2004 Ref-81 Thompson, S.E., C.A. Burton, D.J. Quinn and Y.C. Ng. 1972. Concentration factors of chemical elements in edible aquatic organisms. UCRL-50564. Rev. 1. National Technical Information Service, Springfield, VA. Thompson-Eagle, E.T. and W.T. Frankenberger Jr. 1990. Volatilization of Selenium from Agricultural Evaporation Pond Water. J. Environ. Qual. 19(1):125-131. Thorarinsson, R., M.L. Landolt, D.G. Elliott, R.J. Pascho, and R.W. Hardy. 1994. Effect of dietary vitamin E and selenium on growth, survival and the prevalence of Renibacterium salmoninaruminfection in chinook salmon (

Oncorhynchus tshawytscha). Aquaculture 121(4):343-358. Tian, Y. and F. Liu. 1993. Selenium requirement of shrimp Penaeus chinensis . Chinese J. Oceanol. Limnol. 11(3):249-253. Tilbury, K.L., J.E. Stein, J.P. Meador, C.A. Krone and S.L. Chan. 1997. Chemical contaminants in harbor porpoise ( Phocoena phocoena) from the north Atlantic coast: tissue concentrations and intra- and inter-organ distribution. Chemosphere 34(9/10):2195-2181. Tokunaga, T.K., G.E. Brown, Jr., I.J. Pickering, S.R. Sutton and S. Bajt. 1997. Selenium redox reaction and transport between pondal waters and sediments. Environ. Sci. Technol. 31:1419-1425. Tomasik, P., C.H.D. Magadza, S. Mhizha and A. Chirume. 1995a. The metal-metal interactions in biological systems. Part III. Daphnia magna . Water Air Soil Pollut. 82(3-4):695-711. Tomasik, P., C.M. Magadza, S. Mhizha, A. Chirume, M.F. Zaranyika and S. Muchiriri. 1995b. Metal-metal interactions in biological systems. Part IV. Freshwater snail Bulinus globosus . Water Air Soil Pollut. 83(1-2):123-145. Topcuoglu, S., N. Erenturk, N. Saygi, D. Kut, N. Esen, A. Bassari and E. Seddigh. 1990. Trace metal levels of fish from the Marmara and Black Sea. Toxicol. Environ. Chem. 29(2):95-99. TranVan, L. and D.K. Teherani. 1988. Accumulation and distribution of elements in rice (seed, bran layer, husk) by neutron activation analysis. J. Radioanal. Nucl. Chem. 128(1):35-42. Traversy, W.J., P.D. Goulden, Y.M. Sheikh and J.R. Leacock. 1975. Levels of arsenic and selenium in the Great Lakes region. Scientific Series No. 58. Environment Canada, Burlington, Ontario, Canada. Treuthardt, J. 1992. Hematology Antioxidative Trace Elements The Related Enzyme Actuvities And Vitamin E In Growing Mink On Normal And Anemiogenic Fish Feeding. Acta Acad. Aboensis Ser B Math. Phys. Matematik Natur. Tek. 52(4):1-138. Trieff, N.M., L.A. Romana, A. Esposito, R. Oral, F. Quiniou, M. Iaccarino, N. Alcock, V.M.S. Ramanujam and G. Pagano. 1995. Effluent from bauxite factory induces developmental and reproductive damage in sea urchins. Arch. Environ. Contam. Toxicol. 28(2):173-177. Tripathi, A.K. and S.N. Pandey. 1985. Toxicity of selenium to Chlorella vulgarisand Phormidium foveolarum . Natl. Acad. Sci. Lett. (India) 8(10):307-9.

### Draft November 12, 2004 Ref-82 Trocine, R. P. and J.H. Trefry. 1996. Metal concentrations in sediment, water and clams from the Indian River Lagoon, Florida. Mar. Pollut. Bull. 32(10):754-759. Tsuji, S., Y. Tonogai, Y. Ito and S. Kanoh. 1986. The influence of rearing temperatures on the toxicity of various environmental pollutants for killifish ( Oryzias latipes ). J. Hyg. Chem. Eisei Kagaku 32(1):46-53. Turgeon, D.D. and T.P. O'Connor. 1991. Long Island Sound: distributions, trends, and effects of chemical contamination. Estuaries 14(3):279-289. Turner, M.A. and J.W.M. Rudd. 1983. The English-Wabigoon River system: III. Selenium in lake enclosures: Its geochemistry, bioaccumulation, and ability to reduce mercury bioaccumulation. Can. J. Fish. Aquat. Sci. 40:2228-2240. Turner, M.A. and A.L. Swick. 1983. The English-Wabigoon River system: IV. Interaction between mercury and selenium accumulated from waterborne and dietary sources by northern pike ( Esox lucius ). Can. J. Fish. Aquat. Sci. 40:2241-2250. Twerdok, L.E., D.T. Burton, H.S. Gardner, T.R. Shedd and M.J. Wolfe. 1997. The use of nontraditional assays in an integrated environmental assessment of contaminated ground water. Environ. Toxicol. Chem. 16(9):1816-1820. U.S. EPA. 1976. Quality criteria for water. PB-263943 or EPA-440/9-76-023. National Technical Information Service, Springfield, VA. U.S. EPA. 1978. In-depth studies on health and environmental impacts of selected water pollutants. (Table of data available from C.E. Stephan, U.S. EPA, Duluth, MN.) U.S. EPA. 1980a. Ambient water quality criteria for selenium. EPA-440/5-80-070. National Technical Information Service, Springfield, VA. U.S. EPA. 1980b. Water quality criteria documents. Federal Regist. 45:79318-79379. November 28. U.S. EPA. 1983a. Methods for chemical analysis of water and wastes. EPA-600/4-79-020 (Revised March 1983). National Technical Information Service, Springfield, VA. U.S. EPA. 1983b. Water quality standards regulation. Federal Regist. 48:51400-51413. November 8. U.S. EPA. 1983c. Water quality standards handbook. Office of Water Regulations and Standards, Washington, DC. U.S. EPA. 1985a. Appendix B - Response to public comments on "Guidelines for deriving numerical national water quality criteria for the protection of aquatic organisms and their uses." Federal Regist. 50:30793-30796. July 29. U.S. EPA. 1985b. Water quality criteria. Federal Regist. 50:30784-30792. July 29.

### Draft November 12, 2004 Ref-83 U.S. EPA. 1985c. Technical support document for water quality-based toxics control. EPA-440/4-85-032 or PB86-150067. National Technical Information Service, Springfield, VA. U.S. EPA. 1986. Chapter I - Stream design flow for steady-state modeling. In: Book VI - Design conditions. In: Technical guidance manual for performing waste load allocation. Office of Water, Washington, DC. August. U.S. EPA. 1987a. Ambient water quality criteria for selenium. EPA-440/5-87-006. National Technical Information Service. U.S. EPA. 1987b. Permit writer's guide to water quality-based permitting for toxic pollutants. EPA-440/4- 87-005. Office of Water, Washington, DC. U.S. EPA. 1994. Water quality standards handbook: 2nd ed. EPA-823-B-94-005a,b. Washington, DC. U.S. EPA. 1995. Ambient Water Quality Criteria for selenium. U.S. EPA. 1998. Report on the Peer Consultation Workshop on Selenium Aquatic Toxicity and Bioaccumulation. Office of Water. EPA-822-R-98-007. U.S. EPA. 1999. 1999 Update of Ambient Water Quality Criterion for Ammonia. Uchida, H., Y. Shimoishi and K. Toei. 1980. Gas chromatographic determination of selenium(-II,O), -(IV), and -(VI) in natural waters. Environ. Sci. Technol. 14:541-544. Unsal, M. 1987. Evaluation of the synergistic effect of selenium on the acute toxicity of mercury in fish larvae. Rev. Int. Oceanogr. Med. 87-88(0):125-136. Uthe, J.F. and E.G. Bigh. 1971. Preliminary survey of heavy metal contamination of Canadian freshwater fish. J. Fish. Res. Board Can. 28:786-788. Van Derveer, W.D. and S.P. Canton. 1997. Selenium sediment toxicity thresholds and derivation of water quality criteria for freshwater biota of western streams. Environ. Toxicol. Chem. 16(6):1260-1268. Vandermeulen, J.H. and A. Foda. 1988. Cycling of selenite and selenate in marine phytoplankton. Mar. Biol. (Berlin) 98(1):115-23. Vanderstoep, J., S. Weintraub and K. Barber. 1990. Nutritional Composition of British Columbia Canada Canned Salmon. Can. Inst. Food Sci. Technol. J. 23(2-3):121-124. Van Metre, P.C. and J.R. Gray. 1992. Effects of Uranium Mining Discharges on Water Quality in the Puerco River Basin Arizona and New Mexico. Hydro. Sci. Sci. Hydrol. 37(5):463-480. Van Puymbroeck, S.L.C., W.J.J. Stips and O.L.J. Vanderborght. 1982. The antagonism between selenium and cadmium in a freshwater mollusc. Arch. Environ Contam. Toxicol. 11:103-106. Varanasi, U., J.E. Stein, K.L. Tilbury, J.P. Meador and C.A. Sloan. 1993. Chemical contaminants in gray whales ( Eschrichtius robustus ) stranded in Alaska, Washington, and California, USA. Report,

### Draft November 12, 2004 Ref-84 NOAA-TM-NMFS-NWFSC-11; Order No. PB94-106945, 114 pp. Avail. NTIS From: Gov. Rep. Announce. Index (U. S.) 1994, 94(2), Abstr. No. 405,388. Varanasi, U., J.E. Stein, K.L. Tilbury, J.P. Meador, C.A. Sloan, R.C. Clark and S.L. Chan. 1994. Chemical contaminants in gray whales ( Eschrichtius robustus ) stranded along the west coast of North America. Sci. Total Environ. 145(1-2):29-53. Vazquez, M.S., A.M. Gutierrez, M.M. Gomez and M.A. Palacios. 1994. In vitro study of selenium, copper and zinc absorptible fractions and selenium speciation in mussels. Quim. Anal. (Barcelona), 13(3):144-147. Veena, K.B., C.K. Radhakrishnan and J. Chacko. 1997. Heavy metal induced biochemical effects in an estuarine teleost. Indian J. Mar. Sci. 26(1):74-78. Venables WN, Ripley BD. 2002. Modern applied statistics with S. Springer, New York, NY. Versar. 1975. Preliminary investigation of effects on the environment of boron, indium, nickel, selenium, tin, vanadium and their compounds. Volume IV. Selenium. PB-245987 or EPA-560/2-75-005D. National Technical Information Service, Springfield, VA. Versar. 2000. Peer review of statistical analysis of selenium toxicity data. Report submitted to U.S. EPA, Office of Water, Washington, DC. Viso, A.C., F. Boisson, M. Romeo and M. Gnassia-Barelli. 1989. Combined effects of sulfate, selenium (selenate or selenite) and duration of experiment on a coenocytic alga Bryopsissp. Mar. Environ. Res. 28(1-4):515-519. Vitaliano, J.J. and V.S. Zdanowicz. 1992. Trace metals in eggs of winter flounder from Boston Harbor, a contaminated North American estuary. Mar. Pollut. Bull. 24(7):364-367. Vlieg, P. 1990. Selenium concentration of the edible part of 74 New Zealand fish species. J. Food Compos. Anal. 3(1):67-72. Vlieg, P., T. Murray and D.R. Body. 1993. Nutritional data on six oceanic pelagic fish species from New Zealand waters. J. Food Compos. Anal. 6(1):45-54. Vocke, R.W., K.L. Sears, J.J. O'Toole and R.B. Wildman. 1980. Growth responses of selected freshwater algae to trace elements and scrubber ash slurry generated by coal-fired power plants. Water Res. 14:141- 150. Vos, G., J.P.C. Hovens and P. Hagel. 1986. Chromium, nickel, copper, zinc, arsenic, selenium, cadmium, mercury and lead in Dutch fishery products 1977-1984. Sci. Total Environ. 52(1-2):25-40. Waddell, B. and T. May. 1995. Selenium concentrations in the razorback sucker ( Xyrauchen texanus): Substitution of non-lethal muscle plugs for muscle tissue in contaminant assessment. Arch. Environ. Contam. Toxicol. 28(3):321-326.

### Draft November 12, 2004 Ref-85 Wagemann, R. and F.A.J. Armstrong. 1988. Trace metal determination in animal tissues: An interlaboratory comparison. Talanta 35(7):545-552. Wagemann, R. and R.E.A. Stewart. 1994. Concentrations of heavy metals and selenium in tissues and some foods of walrus (

Odobenus rosmarus ) from the eastern Canadian Arctic and sub-Arctic, and associations between metals, age, and gender. Can. J. Fish. Aquat. Sci. 51(2):426-436. Wagemann, R., R.E.A. Stewart, W.L. Lockhart, B.E. Stewart, and M. Povoledo. 1988. Trace metals and methyl mercury: Associations and transfer in harp seal ( Phoca groenlandica) mothers and their pups. Mar. Mam. Sci. 4(4):339-355. Wagemann, R., S. Innes, and P.R. Richard. 1996. Overview and regional and temporal differences of heavy metals in Arctic whales and ringed seals in the Canadian Arctic. Sci. Total Environ. 186(1-2):41-66. Wahl, C., S. Benson and G. Santolo. 1994. Temporal and spatial monitoring of soil selenium at Kesterson Reservoir, California. Water Air Soil Pollut. 74(3-4):345-361. Walsh, D.F., B.L. Berger and J.R. Bean. 1977. Mercury, arsenic, lead, cadmium, and selenium residues in fish, 1971-1977 - National Pesticide Monitoring Program. Pestic. Monit. J. 11:5-34. Wandan, E.N. and M.J. Zabik. 1996. Assessment of the contamination of surface water and fish from Cote d'Ivoire. J. Environ. Sci. Health, Part B, B31(2):225-240. Wang, C. and R.T. Lovell. 1997. Organic selenium sources, selenomethionine and selenoyeast, have higher bioavailability than an inorganic selenium source, sodium selenite, in diets for channel catfish ( Ictalurus punctatus ). Aquaculture 152(1-4):223-234. Wang, C., R.T. Lovell, and P.H. Klesius. 1997. Response to Edwardsiella ictaloricchallange by channel catfish fed organic and inorganic sources of selenium. J. Aquatic Animal Health 9:172-179. Wang, D., G. Alfthan, A. Aro, A. Maekela, S. Knuuttila and T. Hammar. 1995. The impact of selenium supplemented fertilization on selenium in lake ecosystems in Finland. Agric. Ecosyst. Environ. 54(1-2):137-148. Wang, S., M. Misra, R.G. Reddy and J.C. Milbourne. 1992. Selenium removal from solutions using iota chips. In: Residues Effluents: Process. Environ. Consid., Proc. Int. Symp. Reddy, R.G., W.P. Imrie and P.B. Queneau (Eds). Miner. Met. Mater. Soc.: Warrendale, Pa. pp.757-773. Wang, W. 1986. Toxicity tests of aquatic pollutants by using common duckweed ( Lemna minor). Environ. Pollut. 11B:1-14. Wang, W.X. 1996. Accumulation and retention of trace elements in the mussel, Mytilus edulis(silver, americium, cadmium, cobalt, chromium, selenium, zinc, San Francisco Bay, California, Long Island Sound, New York). 324 pp. Avail. Univ. Microfilms Int., Order No. DA9713847 From: Diss. Abstr. Int., B 1997, 57(11):6834.

### Draft November 12, 2004 Ref-86 Wang, W.X. and N.S. Fisher. 1996a. Assimilation of trace elements and carbon by the mussel Mytilus edulis : Effects of food composition. Limnol. Oceanogr. 41(2):197-207. Wang, W.X. and N.S. Fisher. 1996b. Assimilation of trace elements by the mussel Mytilus edulis : effects of diatom chemical composition. Mar. Biol. (Berlin) 125(4):715-724. Wang, W.X., N.S. Fisher and S.N. Luoma. 1995. Assimilation of trace elements ingested by the mussel Mytilus edulis : Effects of algal food abundance. Mar. Ecol.Prog. Ser. 129(1-3):165-176. Wang, W.X., J.R. Reinfelder, B.-G. Lee and N.S. Fisher. 1996a. Assimilation and regeneration of trace elements by marine copepods. Limnol. Oceanogr. 41(1):70-81. Wang, W.X., N.S. Fisher and S.N. Luoma. 1996b. Kinetic determinations of trace element bioaccumulation in the mussel Mytilus edulis . Mar. Ecol. Prog. Ser. 140(1 to 3):91-113. Ward, D.R. and G.J. Flick. 1990. The effects of salinity and temperature on selected elements in oysters ( Crassostrea virginica). J. Food Compos. Anal. 3(1):96-98. Ward, G.S., T.A. Hollister, P.T. Heitmuller and P.R. Parrish. 1981. Acute and chronic toxicity of selenium to estuarine organisms. Northeast Gulf Sci. 4:73-78. Warren, R.J., B.M. Wallace and P.B Bush. 1990. Trace Elements in Migrating Blue-winged Teal Seasonal Sex and Age-class Variations. Environ. Toxicol. Chem. 9(4):521-528. Watenpaugh, D.E. and T.L. Beitinger. 1985a. Absence of selenate avoidance by fathead minnows (Pimephales promelas). Water Res. 19:923-926. Watenpaugh, D.E. and T.L. Beitinger. 1985b. Oxygen consumption in fathead minnows ( Pimephales promelas) following acute exposure to water-borne selenium. Comp. Biochem. Physiol. 80C:253-256. Watenpaugh, D.E. and T.L. Beitinger. 1985c. Selenium exposure and temperature tolerance of fathead minnows, Pimephales promelas . J. Therm. Biol. 10:83-86. Weast, R.C

. 1969. Handbook of Chemistry and Physics, 50th Ed . Chemical Rubber Company Cleveland, OH. Weber, O. 1985. Concentrations of metals in fish from the River Rednitz. Z. Lebensm. Unters. Forsch. 180:463-466. Wehr, J.D. and L.M. Brown. 1985. Selenium requirement of a bloom-forming planktonic alga from softwater and acidified lakes. Can. J. Fish. Aquat. Sci. 42:1783-1788. Weir, P.A. and C.H. Hine. 1970. Effects of various metals on behavior of conditioned goldfish. Arch. Environ. Health 20:45-51. Welsh, D. 1992. Selenium in aquatic habitats at Cibola National Wildlife Refuge. Avail. Univ. Microfilms Int., Order No. DA9309028 From: Diss. Abstr. Int. B 1993, 53(8):5626.

### Draft November 12, 2004 Ref-87 Welsh, D. and O.E. Maughan. 1994. Concentrations of selenium in biota, sediments, and water at Cibola National Wildlife Refuge. Arch. Environ. Contam. Toxicol. 26(4):452-458. Wen, H.Y., R.L. Davis, B. Shi, J.J. Chen, L. Chen, M. Boylan, and J.E. Spallholz. 1997. Bioavailability of selenium from veal, chicken, beef, pork, lamb, flounder, tuna, selenomethionine, and sodium selenite assessed in selenium-deficient rats. Biol. Trace Elem. Res. 58:43-53. Wenzel, C. and G.W. Gabrielsen. 1995. Trace element accumulation in three seabird species from Hornoya, Norway. Arch. Environ. Contam. Toxicol. 29(2):198-206. Weres, O., H.R. Bowman, A. Goldstein, E.C. Smith and L. Tsao. 1990. The Effect of Nitrate and Organic Matter upon Mobility of Selenium in Groundwater and in a Water Treatment Process. Water Air Soil Pollut. 49(3-4):251-272. Westerman, A.G. and W.J. Birge. 1978. Accelerated rate of albinism in channel catfish exposed to metals. Prog. Fish-Cult. 40:143-146. Wheeler, A.E., R.A. Zingaro, K. Irgolic and N.R. Bottino. 1982. The effect of selenate, selenite, and sulfate on the growth of six unicellular marine algae. J. Exp. Mar. Biol. Ecol. 57:181-194. White, D.H. and J.G.H. Geitner. 1996. Environmental contaminants and productivity in an extinct heronry at Charleston Harbor, South Carolina, U.S.A. 1984. Environ. Monitor. Assess. 40(2):137-141. Whyte, J.N.C. and J.A. Boutillier. 1991. Concentrations of inorganic elements and fatty acids in geographic populations of the spot prawn Pandalus platyceros

. Can. J. Fish. Aquat. Sci. 48(3):382-390. Wiemeyer, S.N. and D.J. Hoffman. 1996. Reproduction in eastern screech-owls fed selenium. J. Wildl. Manage. 60(2):332-341. Wiemeyer, S.N., R.M. Jurek and J.F. Moore. 1986. Environmental contaminants in surrogates, foods and feathers of California condors ( Gymnogyps californianus ). Environ. Monitor. Assess. 6(1):91-111. Wilber, C.G. 1980. Toxicology of selenium: A review. Clin. Toxicol. 17:171-230. Wilber, C.G. 1983. Selenium: A potential environmental poison and a necessary food constituent. Charles C. Thomas Publishing Company, Springfield, IL. Wildhaber, M.L. and C.J. Schmitt. 1996. Hazard ranking of contaminated sediments based on chemical analysis, laboratory toxicity tests, and benthic community composition: prioritizing sites for remedial action. J. Great Lakes Res. 22(3):639-652. Williams, M.J., R.S. Ogle, A.W. Knight and R.G. Burau. 1994. Effects of sulfate on selenate uptake and toxicity in the green algaSelenastrum capricornutum . Arch. Environ. Contam. Toxicol. 27(4):449-453. Williams, M.L., R.L. Hothem and H.M Ohlendorf. 1989. Recruitment Failure in American Avocets and Black-necked Stilts Nesting at Kesterson Reservoir California USA 1984-1985. Condor 91(4):797-802.

### Draft November 12, 2004 Ref-88 Williams, T.P., J.M. Bubb and J.N. Lester. 1994. The occurrence and distribution of trace metals in halophytes. Chemosphere 28(6):1189-1199. Wilson, D.S., P. Zhang, R. He, R. Ota and S.T. Omaye. 1996. Comparative distribution of selenoproteins in tissues of female rats and Mallard ducks injected with selenium-75. Toxic Subst. Mech. 15(4):343-354. Wilson, D.S., P. Zhang, R. He, R. Ota, and S.T. Omaye. 1997. Kinetics of selenium incorporation into tissues of female mallard ducks. Toxicol. 122:51-60. Wilson, E.A., E.N. Powell, T.L. Wade, R.J. Taylor, B.J. Presley and J.M. Brooks. 1992. Spatial and Temporal Distributions of Contaminant Body Burden and Disease in Gulf of Mexico Oyster Populations the Role of Local and Large-scale Climatic Controls. Helgol. Wiss. Meeresunters. 46(2):201-235. Wilson, R.D., P.R. Bowser and W.E. Poe. 1984. Dietary vitamin E requirement of fingerling channel catfish. J. Nutr. 114:2053-2058. Winger, P.V. and J.K. Andreasen. 1985. Contaminant residues in fish and sediments from lakes in the Atchafalaya River basin (Louisiana). Arch. Environ. Contam. Toxicol. 14:579-586. Winger, P.V., C. Sieckman, T.W. May and W.W. Johnson. 1984. Residues of organochlorine insecticides, polychlorinated biphenyls, and heavy metals in biota from Apalachicola River, Florida. 1978. J. Assoc. Off. Anal. Chem. 67:325-333. Winger, P.V., D.P. Schultz and W.W. Johnson. 1990. Environmental Contaminant Concentrations in Biota from the Lower Savannah River Georgia and South Carolina USA. Arch. Environ. Contam. Toxicol. 19 (1):101-117. Winner, R.W. 1984. Selenium effects on antennal integrity and chronic copper toxicity in Daphnia pulex (deGeer). Bull. Environ. Contam. Toxicol. 33:605-611. Winner, R.W. 1989. Multigeneration life-span tests of the nutritional adequacy of several diets and culture waters for Cerio daphnia dubia . Environ. Toxicol. Chem. 8(6):513-520. Winner, R.W. and T.C. Whitford. 1987. The interactive effects of a cadmium stress, a selenium deficiency and water temperature on the survival and reproduction of Daphnia magnaStraus. Aquat. Toxicol. (Amsterdam) 10(4):217-224. Wise, D.J., J.R. Tomasso, D.M. Gatlin III, S.C. Bai and V.S. Blazer. 1993a. Effects Of Dietary Selenium And Vitamin E On Red Blood Cell Peroxidation Glutathione Peroxidase Activity And Macrophage Superoxide Anion Production In Channel Catfish. J. Aquat. Animal Health 5(3):177-182. Wise, S.A., M.M. Schantz, B.J. Koster, R. Demiralp, E.A. Mackey, R.R. Greenberg, M. Burow, P. Ostapczuk and T.I. Lillestolen. 1993b. Development of frozen whale blubber and liver reference materials for the measurement of organic and inorganic contaminants. Fresenius. Anal. Chem. 345(2-4):270-277. Wolfe, D.A., E.R. Long and G.B. Thursby. 1996. Sediment toxicity in the Hudson-Raritan estuary: distribution and correlations with chemical contamination. Estuaries 19(4):901-912.

### Draft November 12, 2004 Ref-89 Wolfenberger, V. 1986. Survival of the hermit crab, Clibanarius vittatus , exposed to selenium and other environmental factors. Bull. Environ. Contam. Toxicol. 37(3):369-74. Wolfenberger, V.A. 1987. Influence of environmental factors on oxygen consumption of Clibanarius vittatus(striped hermit crab). Texas J. Sci. 39(1):37-48. Wong, D. and L. Oliveira. 1991a. Effects of selenite and selenate on the growth and motility of seven species of marine microalgae. Can. J. Fish. Aquat. Sci. 48(7):1193-1200. Wong, D. and L. Oliveira. 1991b. Effects of selenite and selenate toxicity on the ultrastructure and physiology of three species of marine microalgae. Can. J. Fish. Aquat. Sci. 48(7):1201-1211. Wong, P.T.S. and Y.K. Chau. 1988. Toxicity of metal mixtures to phytoplankton. In: Heavy Met. Hydrol. Cycle. Astruc, M., J.N. Lester (Eds). Selper Ltd.: London, UK. pp. 231-236. Wong, P.T.S., Y.K. Chau and D. Patel. 1982. Physiological and biochemical responses of several freshwater algae to a mixture of metals. Chemosphere 11:367-376. Woock, S.E. and P.B. Summers, Jr. 1984. Selenium monitoring in Hyco Reservoir (NC) waters (1977- 1981) and biota (1977-1980). In: Workshop proceedings: The effects of trace elements on aquatic ecosystems. EA-3329. Electric Power Research Institute, Palo Alto, CA. pp. 6-1 to 6-27. Woock, S.E., W.R. Garrett, W.E. Partin, and W.T. Bryson. 1987. Decreased survival and teratogenesis during laboratory selenium exposures to bluegill, Lepomis macrochirus . Bull. Environ. Contam. Toxicol. 39(6):998-1005. Wren, C.D. and H.R. Maccrimmon. 1986. Comparative bioaccumulation of mercury in two adjacent freshwater ecosystems. Water Res. 20(6):763-770. Wren, C.D., P.M. Stokes and K.L. Fischer. 1987. Mercury levels in Ontario (Canada) mink and otter relative to food levels and environmental acidification. Can. J. Zool. 64(12):2854-2859. Wrench, J.J. 1978. Selenium metabolism in the marine phytoplankters Tetraselmis tetratheleand Dunaliella minuta . Mar. Biol. (Berl.) 49:231-236. Wrench, J.J. 1979. Uptake and metabolism of selenium by oysters. Mar. Sci. Commun. 5:47-59. Wrench, J.J. and C.I. Measures. 1982. Temporal variations in dissolved selenium in a coastal ecosystem. Nature 299:431-433. Wu, L. and Z.Z. Huang. 1991. Selenium Accumulation and Selenium Tolerance of Salt Grass from Soils with Elevated Concentrations of Selenium and Salinity. Ecotoxicol. Environ. Safety 22(3):267-282. Wu, L., A.W. Enberg and X. Guo. 1997. Effects of elevated selenium and salinity concentrations in root zone on selenium and salt secretion in saltgrass ( Distichlis spicataL.). Ecotoxicol. Environ. Safety 37(3):251-258.

### Draft November 12, 2004 Ref-90 Yamamoto, T., Y. Otsuka, K. Aoyama, H. Tabata,and K. Okamoto. 1984. The distribution of chemical elements in selected marine organisms: comparative biogeochemical data. Mar. Estauarine Geochem., [Proc. Symp.], Meeting Date 1984, 315-27. Editor(s): Sigleo, Anne C.; Hattori, Akihiko. Lewis Publ., Inc.: Chelsea, Mich. Yamaoka, Y., O. Takimura and H. Fuse. 1994. Effects of various elements on arsenic accumulation of the Alga Dunaliella salina . Appl. Organomet. Chem. 8(3):229-235. Yamaoka, Y., O. Takimura, H. Fuse, K. Kamimura and K. Murakami. 1996. Accumulation of arsenic by Rhaphidophyceae Chattonella antiqua (Hada) Ono. Appl. Organom, et. Chem. 10(9):721-726. Yamazaki, M., Y. Tanizaki and T. Shimokawa. 1996. Silver and other trace elements in a freshwater fish, Carassius auratus , from the Asakawa River in Tokyo, Japan. Environ. Pollut. 94(1):83-90. Yan, L., and G.D. Frenkel. Effect of Selenite on Cell Surface Fibronectin Receptor. Biological Trace Element Research. 46:79- . Yokota, A., S. Shigeoka, T. Onishi, and S. Kitaoka. 1988. Selenium as inducer of glutathione peroxidase in low carbon dioxide grownChlamydomonas reinhardtii . Plant Physiol. (Bethesda) 86(3):649-651. Yoshida, M. and K. Yasumoto. 1987. Selenium contents of rice grown at various sites in Japan. J. Food Comp. Anal. 1(1):71-75. Yoshii, O., K. Hiraki, Y. Nishikawa, and T. Shigematsu. 1977. Fluorometric determination of selenium(IV) and selenium(VI) in sea water and river water (in Japanese). Bunseki Kagaku. 26:91- . Yu, R., J.P. Coffman, V. Van Fleet-Stalder and T.G. Chasteen. 1997. Toxicity of oxyanions of selenium and of a proposed bioremediation intermediate, dimethyl selenone. Environ. Toxicol. Chem. 16(2):140-145. Yurkowski, M. 1986. Suitability of two rainbow trout ( Salmo gairdneri ) reference diets for Arctic charr (Salvelinus alpinus ). Can. Tech. Report Fish. Aquat. Sci. 0(1464): I-IV, 1-10. Zagatto, P.A., E. Gherardi-Goldstein, E. Bertoletti, C.C. Lombardi, M.H.R.B. Martins and M.L.L.C.Ramos. 1987. Bioassays with aquatic organisms: toxicity of water and sediment from Cubatao River basin. Water Sci. Technol. 19(11, Eff. Contam. Ecol. Syst.):95-106. Zaidi, J.H., I.H. Qureshi, M. Arif and I. Fatima. 1995. Trace elements determination in some species of fish commonly consumed in Pakistan. Int. J. Environ. Anal. Chem. 60(1):15-22. Zar, J.H. 1984. Biostatiscal Analysis.Prentice-Hall, Englewood-Cliffs, New Jersey, USA. Zar, J. H. 1999. Biostatistical analysis, Fourth edition. Prentice-Hall, Upper Saddle River, NJ. Zatta, P., P. Buso and G. Moschini. 1985. Selenium distribution in the tissues of Carcinus maenas . Comp. Biochem. Physiol. 81C:469-470.

### Draft November 12, 2004 Ref-91 Zawislanski, P.T., and A.E. McGrath. 1998. Selenium Cycling in Estuarine Wetlands: Overview and New Results from the San Francisco Bay. In: W.T. Frankeberger and R.A. Engberg (eds.), Environmental Chemistry of Selenium . Marcel Dekker, New York. pp. 223-242. Zeisler, R., S.F. Stone and R.W. Sanders. 1988. Sequential determination of biological and pollutant elements in marine bivalves. Anal. Chem. 60(24):2760-5. Zeisler, R., R. Demiralp, B.J. Koster, P.R. Becker, M. Burow, P. Ostapczuk and S.A. Wise. 1993. Determination of inorganic constitutents in marine mammal tissues. Sci. Total Environ. 139-140:365-386. Zhang, Y. and J.N. Moore. 1996. Selenium fractionation and speciation in a wetland system. Environ. Sci. Technol. 30:2613-2619. Zhang, G.H., M.H. Hu, Y.P. Huang and P.J. Harrison. 1990. Selenium uptake and accumulation in marine phytoplankton and transfer of selenium to the clam Puditapes philippnarum . Mar. Environ. Res. 30(3):179-190. Zhang, Y. and J.N. Moore. 1997. Environmental conditions controlling selenium volatilization from a wetland system. Environ. Sci. Tech. 31(2):511-517. Zhang, Yuanxun, R. Moro, and G. Gialanella. 1996. Toxic effects of selenium on marine fish. J. Environ. Sci. (China) 8(2):151-156. Zhou, H. and J. Liu. 1997. The simultaneous determination of 15 toxic elements in foods by ICP-MS. At. Spectrosc. 18(4):115-118.