DRAFT

ALBERTA WATER QUALITY GUIDELINE FOR THE PROTECTION OF

FRESHWATER AQUATIC LIFE

COPPER

August 1996

Standards and Guidelines BranchEnvironmental Assessment Division

Environmental Regulatory Service

The water quality guideline derived in this document protects freshwater aquatic life againstcopper toxicity. This guideline is based on up-to-date, high quality scientific information and isderived according to the Protocol to Develop Alberta Water Quality Guidelines for Protection ofFreshwater Aquatic Life (Alberta Environmental Protection 1996). Using the Protocol documentwith this guideline document will place the copper guideline in context.

This document has been reviewed within Alberta Environmental Protection. However, werecognize that other information may exist that could have a bearing on this document. For thisreason alone, this document is called "Draft". Comments from the public are invited untilSeptember 30, 1997. After this period, the comments will be addressed in a "Final" document.

Any comments, questions, or suggestions regarding the content of this document may be directedto:

Standards and Guidelines BranchAlberta Environmental Protection6th Floor, 9820-106 StreetEdmonton, Alberta T5K 2J6Phone: (403) 422-6102

Additional copies of the Alberta Water Quality Guideline for the Protection of FreshwaterAquatic Life - Copper may be obtained by contacting :

Regulatory Approvals CentreAlberta Environmental ProtectionMain Floor, 9820-106 StreetEdmonton, Alberta T5K 2J6Phone: (403) 427-6311

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EXECUTIVE SUMMARY

This document contains the relevant toxicological information and other literature used todevelop a water quality guideline that protects freshwater aquatic life against copper toxicity. Itsderivation follows the Protocol to Develop Alberta Water Quality Guidelines for Protection ofFreshwater Aquatic Life (Alberta Environmental Protection 1996).

Copper is an essential compound for plants and animals in small quantities. However, copperbecomes toxic when biological requirements are exceeded. Copper is widely distributed inenvironmental media because it is a naturally occurring element. Compared to natural emissions,emissions of copper from human activities are substantial. Most copper released from humanactivities comes from disposal of coal ash residue and spreading of municipal and industrialwastes on land.

Cupric ion is the main toxic form of copper. Cupric ion in water is bound (complexed) withinorganic and organic compounds, which reduces cupric ion concentrations (and its toxicity)substantially. The effects of copper on freshwater organisms are numerous. This documentdescribes physiological effects, behavioral effects, acute toxicity (lethal effects), chronic toxicity(long-term effects), microcosm and ecosystem effects, and genotoxic effects. This documentalso presents a water quality guideline based on the acute toxic effects, chronic toxic effects andgenotoxic effects of copper.

High quality (primary) information on short-term effects (acute toxicity) of copper was availablefor 29 fish species (five different orders), 24 invertebrate species (11 different orders) and oneamphibian species. According to the Protocol to Develop Alberta Water Quality Guidelines forProtection of Freshwater Aquatic Life (Alberta Environmental Protection 1996), the minimumrequirements for developing an acute guideline were amply met. The acute toxicity informationon three species and one invertebrate species clearly indicated that acute copper toxicitydiminished in water with increasing hardness. This relationship was included in deriving anacute copper guideline equation.

Primary data on long-term effects (chronic toxicity) of copper were available for 11 fish species(from four different orders), eight invertebrate species (form five different orders), and five plantspecies. According to the Protocol to Develop Alberta Water Quality Guidelines for Protectionof Freshwater Aquatic Life (Alberta Environmental Protection 1996), the minimum requirementsfor developing a chronic guideline were amply met. The chronic toxicity information did notindicate that chronic toxicity of copper was affected by water hardness.

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The resulting Alberta water quality guideline that protects freshwater aquatic life against coppertoxicity is:

Copper Guideline (mg/L)

Acute e0.979123*ln(hardness)-8.64497

Chronic 0.0071

1 The chronic toxicity of copper in soft water was inconclusive: the chronic guideline cantherefore only be applied at water hardness equal to or greater than 50 mg/L CaCO3.

Both acute and chronic copper guidelines apply to acid-extractable copper concentrations. Insurface water, many substances are present that bind copper: the amount of copper that could betoxic is thereby greatly diminished. Specific guidance in this document should be used toaccurately apply the copper guideline. Additional guidance is provided in the Protocol toDevelop Alberta Water Quality Guidelines for Protection of Freshwater Aquatic Life (AlbertaEnvironmental Protection 1996).

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TABLE OF CONTENTS

EXECUTIVE SUMMARY . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . i

TABLE OF CONTENTS . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . iii

1 BACKGROUND INFORMATION ON COPPER . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 11.1 Physical and Chemical Properties . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 11.2 Method of Analysis & Current Detection Limits . . . . . . . . . . . . . . . . . . . . . . . . . . 11.3 Production and Uses . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 31.4 Sources to the Environment . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 41.5 Environmental Concentrations . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 6

2 ENVIRONMENTAL FATE AND BEHAVIOUR . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 82.1 Oxidation states . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 82.2 Inorganic Complexation . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 82.3 Organic Complexation . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 92.4 Adsorption . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 11

3 ENVIRONMENTAL EFFECTS OF COPPER . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 123.1 Effects of Copper on Fish . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 123.2 Effects of Copper on Invertebrates . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 173.3 Effects of Copper on Amphibians . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 213.4 Effects of Copper on Plants . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 213.5 Effects of Copper on Other Microorganisms . . . . . . . . . . . . . . . . . . . . . . . . . . . . 233.6 Effects of Copper on Microcosms and Ecosystems . . . . . . . . . . . . . . . . . . . . . . . 243.7 Genotoxicity of Copper . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 273.8 Metal-binding Proteins (Biomarker) . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 28

4 DERIVATION OF ACUTE AND CHRONIC GUIDELINES . . . . . . . . . . . . . . . . . . . . 294.1 Final Acute Value and Alberta Acute Guideline . . . . . . . . . . . . . . . . . . . . . . . . . 294.2 Alberta Chronic Guideline . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 334.3 Application of the Alberta Copper Guideline . . . . . . . . . . . . . . . . . . . . . . . . . . . 38

5 DATA GAPS . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 41

6 COPPER GUIDELINES FROM OTHER JURISDICTIONS . . . . . . . . . . . . . . . . . . . . . 42

7 REFERENCES . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 44

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APPENDIX 1. Acute and chronic copper toxicity data on amphibians . . . . . . . . . . . . . . 76APPENDIX 2. Copper toxicity data on freshwater algae and plants . . . . . . . . . . . . . . . . 78APPENDIX 3. Acute copper toxicity data on freshwater fish . . . . . . . . . . . . . . . . . . . . . 84APPENDIX 4. Acute copper toxicity data on freshwater invertebrates . . . . . . . . . . . . . . 95APPENDIX 5. Computations for the acute Alberta copper guideline . . . . . . . . . . . . . . 102APPENDIX 6. Chronic copper toxicity data on freshwater fish . . . . . . . . . . . . . . . . . . 114APPENDIX 7. Chronic copper toxicity data on freshwater invertebrates . . . . . . . . . . . 118APPENDIX 8. Computations for the chronic Alberta copper guideline . . . . . . . . . . . . 122

List of Tables

Table 1. Physical and chemical properties of metallic copper . . . . . . . . . . . . . . . . . . . . . . . 2Table 2. Analytical methods for determining copper in water and wastewater . . . . . . . . . . 3Table 3. Worldwide emissions of copper (103 t/yr) into the environment at the beginning of

the 1980s . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 5Table 4. Statistical information on regression and ancova analyses (Sokal and Rohlf 1981)

for derivation of an acute copper guideline. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 30Table 5. Statistical information on regression analyses for a chronic guideline . . . . . . . . 33Table 6. Primary acute-to-chronic ratios (ACR) from copper toxicity studies. . . . . . . . . . 34Table 7. Copper guidelines for the protection of freshwater aquatic life in other

jurisdictions. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 42Table 8. Comparison of freshwater aquatic life guidelines for copper (in :g/L) from

various jurisdictions. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 43

List of Figures

Figure 1. Primary copper GMAVs (Genus Mean Acute Values) . . . . . . . . . . . . . . . . . . . . 32Figure 2. Correlation between chronic copper toxicity and water hardness for fathead

minnow . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 34Figure 3. Primary copper toxicity studies: acute toxicity versus acute-to-chronic ratio

(ACR) . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 35Figure 4. Primary copper GMCVs (Genus Mean Chronic Values) . . . . . . . . . . . . . . . . . . 36

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1 BACKGROUND INFORMATION ON COPPER

Copper is considered to be an essential trace element for plants and animals; it is a component ofmany metalloenzymes and respiratory pigments (Demayo and Taylor 1981). It is required in thesynthesis of chlorophyll (a photosynthetic pigment in plants) and haemoglobin (a respiratoryblood pigment). Copper also serves as the oxygen coupling site in haemocyanin, the respiratoryblood pigment in many molluscs and certain other invertebrates (Birge and Black 1979). Copperis important for maintaining optimum plant metabolism. Copper deficiency results in reducedsynthesis of the copper-containing electron carriers plastocyanin and cytochrome oxidase. Thereduction of these electron carriers reduces photosynthesis and respiration (Barón et al. 1995). However, copper becomes toxic to aquatic biota when biological requirements are exceeded.

1.1 Physical and Chemical Properties

Copper (Chemical Abstracts Service CAS Registry Number 7440-50-8) is classified as a noblemetal (as are silver and gold) and can be found in nature in the elemental form. Its chemical andphysical properties include high thermal conductivity, high electrical conductivity, malleability,low corrosion, alloying ability, and aesthetically pleasing appearance. These properties make itone of the most important metals (U.S. Department of Health & Human Services 1990). Thephysical and chemical properties of copper are presented in Table 1.

1.2 Method of Analysis & Current Detection Limits

Metals may be determined satisfactorily by atomic absorption spectrometry (AAS), inductivelycoupled plasma atomic emission spectrometry (ICP-AES), inductively coupled plasma massspectrometry (ICP-MS), or, with somewhat less precision and sensitivity, by colorimetricmethods (Standard Methods 1992; Long and Martin 1992). The various methods are listed inTable 2 with information on typical instrumental detection limits and analytical ranges. Thesemethods do not permit distinction between different chemical forms or species of copper, unlessthey are combined with various separation techniques such as ion chromatography, highperformance liquid chromatography, gas chromatography, flow injection technique,electrochemical techniques, etc. Among electrochemical techniques, the use of copper ionelectrode potentiometry permits determination of the hydrated cupric Cu(II) ion; however, it isnot accurate at concentrations < 5 :g/L. Both anodic stripping voltammetry (ASV) anddifferential pulse ASV determine copper complexes of relatively low stability (Spear and Pierce1979).

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Table 1. Physical and chemical properties of metallic copper (from U.S. Department ofHealth & Human Services 1990).

Property Copper

Atomic/Molecular Weight 63.546

Color reddish

Physical State solid

Melting Point oC 1083.4

Boiling Point oC 2567

Density g/cm3 8.92

Odour Odour Threshhold: Water/Air

None No data

Taste Taste Threshhold

No data No data

Solubility Water (g/100mL) Organic Solvents

insoluble (CuSO4 14.3 @ 0oC)" (CuSO4 soluble in methanol, slightlysoluble in ethanol)

Vapour pressure, mm Hg 10 (1870 oC)

Autoignition temperature No data

Different copper fractions in water may be reported:(a) dissolved copper, copper in an unacidified sample that passes through a 0.45 :m

membrane filter;(b) total copper, copper in an unfiltered sample after vigorous digestion with a concentrated

acid, or the sum of dissolved and suspended copper;(c) acid-extractable copper, copper in an unfiltered sample after treatment with a hot dilute

mineral acid or after the addition of a dilute mineral acid; and,(d) suspended/residue copper, total copper in an unacidified sample retained by a 0.45 :m

membrane filter.

In Alberta, dissolved, total and acid-extractable copper is measured by graphite furnace AAS(detection limit of 0.001 mg/L), by ICP-AES (detection limit 0.001 mg/L with 5-fold pre-concentration), and by ICP-MS (detection limit 0.00005 mg/L).

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Table 2. Analytical methods for determining copper in water and wastewater (from Mancyand Allen 1977; Standard Methods 1992; Long and Martin 1992).

Instrument Detection Limit (mg/L)

Optimum Range/Upper Limit (mg/L)

Atomic absorption

Flame AAS 0.01 0.2-10

Electrothermal AAS 0.001* 0.005-0.1

Plasma emission

ICP-AES 0.006 upper limit 50

ICP-MS 0.00003 0.0005-0.5

Electrochemical

Anodic strippingvoltammetry

0.00006-0.006

Colorimetric

Neocuproine 0.0006

Bathocuproine 0.020* combined with flow injection

1.3 Production and Uses

In Canada, 35 mines produced copper as the main product or one of the products and threeadditional mines produced copper as a by-product during 1990 (Environment Canada 1992). In1990 and 1991, three of these mines closed and one suspended operations. The copper mines arelocated in British Columbia, Manitoba, Ontario, Quebec and New Brunswick. Rated millingcapacity of mines with copper as a main product varied from 330 tonnes of ore per day (tpd) to140,000 tpd in 1990. These data indicated that most of the production is in British Columbia(61.8%) and Ontario (24.4%). Total production in 1990 was similar to the production in 1975(Wood 1976, as quoted in Demayo and Taylor 1981). Operations of these non-ferrouscompanies concentrate on smelting and refining, and are largely export oriented (IndustryCanada 1994).

The metal copper and its compounds have been used by man since prehistoric times because ofits properties such as malleability, ductility, conductivity, corrosion resistance, alloying qualities,and pleasing appearance. The main uses today are in the electrical, construction, plumbing, andautomotive industries. The electrical industry is the largest single user (>50% of total

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production) and covers power transmission, electronics and electrical equipment. Constructionand plumbing is the second largest user of copper (piping and the manufacture of alloys [e.g.,bronze, brass]). Copper sulfate and cuprous oxude have been used as fungicides for manydecades. However, the use of copper as a biocide has decreased considerably (Demayo andTaylor 1981; Moore and Ramamoorthy 1984).

1.4 Sources to the Environment

Copper and its compounds are naturally present in the earth's crust. Natural discharges to air andwater may therefore be significant. The latest worldwide emission figures for various metals(including copper) are for the beginning of the 1980s (summarized by Pacyna et al. 1995; Table3). Most of the copper released to the environment is released to soil and least is released to air(Table 3).

Releases to Air

On a global scale, natural and anthropogenic copper emissions to air are similar in magnitude. Principal natural sources are wind-borne soil particles and volcanoes. The main anthropogenicsource is emissions from the primary non-ferrous metal industry (Pacyna et al. 1995). In theUnited States, copper emissions to air are estimated to be only 0.4% of copper released to theenvironment with windblown dust as the primary natural source (U.S. Department of Health &Human Services 1990).

Releases to Water

Both natural and anthropogenic sources contribute copper to water: natural weathering of soil,atmospheric deposition, and discharges from industry and wastewater treatment plants. A 1976evaluation in the United States determined that 2.4% of the identified copper releases to theenvironment enters waterways. The major source is from land runoff through natural weathering(68%; U.S. Department of Health & Human Services 1990). Copper sulfate use represented 13%of the releases to water; urban runoff contributed 2% (Perwack et al. 1980 as quoted in U.S.Department of Health & Human Services 1990).

Releases to Soil

On a global scale, the two principal sources of copper to soils are disposal of ash residues fromcoal combustion and municipal and industrial wastes on land. Mine tailing and slags and wastesfrom smelters can also be a major source of copper to soils: 657-1577 103 t/year (Nriagu andPacyna 1988).

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Table 3. Worldwide emissions of copper (103 t/yr) into the environment at the beginning ofthe 1980s (after Pacyna et al. 1995).

Sources 103 t/yr Subtotal Total

Air Natural windborn soil particles 0.9-15.0

2.2-53.8

21.9-104.6

seasalt spray 0.2-6.9

volcanoes 0.9-18.0

wild forest fires 0.1-7.5

biogenic processes 0.1-6.4

Anthropogenic non-ferrous metalindustry

15.2-35.4

19.7-50.8fuel combustion 2.8-11.4

other industry and use 0.6-1.9

waste incineration 1.1-2.1

Water Anthropogenic waste disposal 11.6-70.0

34.7-190.5

steam electric 3.6-23.0

mining,smelting,refining

2.5-26.0

manufacturingprocesses

11.0-56.5

atmospheric deposition 6.0-15.0

Soil Anthropogenic disposal municipal andindustrial waste

39.2-239.2

541.5-1402.8

wastage of commercialproducts

395.0-790.0

coal fly ash and bottomfly ash

93.0-335.0

fertilizer 0.1-0.6

peat 0-0.2

atmospheric deposition 14.0-36.0

In the United States, an estimated 97% of copper released into the environment is to land. Tailings and overburdens from copper mines and tailings from mills are the primary source. Agricultural products constitute 2% of the copper released to soil (Perwack et al. 1980 as quotedin U.S. Department of Health & Human Services 1990).

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Canadian Copper Releases

A 1993 summary of copper (and its compounds) releases in Canada is provided by NPRI(National Pollutant Release Inventory). This inventory includes all facilities that manufacture,process or otherwise use any of the NPRI substances in quantities of 10 tonnes or more per year,employ 10 or more people or manufacture, and process or otherwise used the substance in greaterthan 1% weight. For those facilities reporting to NPRI, 32 facilities released more than 1.0 tonneper year in 1993. In total, these facilities released 13,998 tonnes of copper per year. Of this total,85% was released to water from a copper mine in British Columbia (11,890 tonnes per year). The copper releases were 38.9% to land (landfarm and landfill), 29.4% to water and 31.4% to air(stack, storage and fugitive). Only one of the 32 facilities is located in Alberta (Sherritt Inc., FortSaskatchewan): their total release of copper in 1993 was 30.1 tonnes, most (99%) of it waslandfarmed.

Although facilities engaged in mining are not subject to reporting their releases to NPRI, coppermines are subject to the Metal Mining Liquid Effluent Regulations or associated guidelines. Theauthorized numerical levels for copper in the regulations and guidelines are 0.3 mg/L maximummonthly arithmetic mean, 0.45 mg/L maximum concentration in a composite sample, and 0.6mg/L maximum concentration in a grab sample. One of the 35 copper mines was not subject tothe regulations or guidelines. Of the remaining 34 mines, 7 occasionally exceeded theregulations or guidelines (Environment Canada 1992). Alberta has no mines that produce copperas a main product or a by-product.

1.5 Environmental Concentrations

Soil

The crustal average concentration of copper has been estimated at 55 mg/kg but varies with thetype of rock (10 mg/kg for granite and 100 mg/kg for basalt). Copper ranks 25th in abundanceamong the elements present in the earth's crust. Copper concentrations in soils range between 2and 100 mg/kg dry weigth with a mean value of 20 mg/kg (Bowen 1966 as quoted in Demayoand Taylor 1981).

Sediment

Mean copper concentrations in freshwater sediments ranged from 12 to 57 mg/kg with individualvalues as high as 4000 mg/kg (Spear and Pierce 1979). Copper concentrations in pristineenvironments are generally less than 50 mg/kg dry weigth, while concentrations in pollutedenvironments can be several thousand mg/kg (Harrison and Bishop 1989).

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Copper concentrations in sediments of the Battle River (Alberta) varied from 1.16 to 35.5 mg/kg(0.5N HCl-extractable). The highest copper concentrations in sediments were found in reservoirsand sites with high levels of organic matter (Anderson et al. 1994).

Freshwater

Copper is widely distributed in water since it is a naturally occurring element. Copper levels inriver waters range from 0.6 to 400 :g/L, with a median of 10 :g/L. Dissolved copper levels inuncontaminated freshwaters usually range from 0.5 to 1.0 :g/L, increasing to > 2 :g/L in urbanareas (Moore and Ramamoorthy 1984). Dissolved copper levels in Canadian surface watersrarely exceed 5 :g/L (Spear and Pierce 1979). At 11 interprovincial border sites in the prairies,total copper concentrations varied from below detection to 85 :g/L; most median values werearound 3 :g/L with some values as high as 8 :g/L (PPWB 1993). Similar median values fortotal copper are reported for Federal-Provincial monitoring sites in British Columbia (Singleton1987).

In Alberta rivers, copper concentrations are generally near detection limit. In the Peace River,mean total concentrations increased from 0.002 mg/L at the B.C.-Alberta border to 0.013 mg/Lbefore the Peace River enters Lake Athabasca. Dissolved copper was on average 25% of totalcopper. High copper levels were largely a result of increased suspended solid concentrations(Shaw et al. 1990). In the Athabasca River, no spatial trends in total copper were observed. Most total copper concentrations were around the detection limit (0.001 mg/L); the highestobserved value was 0.032 mg/L (Noton and Shaw 1989; Noton and Saffran 1995). High coppervalues were generally observed in spring, associated with high concentrations of suspendedsolids (Noton and Saffran 1995). In the North Saskatchewan River, copper concentrations weregenerally higher downstream of the City of Edmonton than upstream. The increase was due tonatural and anthropogenic copper loads (Shaw et al. 1994). In the Red Deer River, medianconcentrations of total and extractable copper varied from 0.001 to 0.004 mg/L and 0.001 to0.003 mg/L, respectively (Shaw and Anderson 1994). In the Bow River, copper concentrationsoccasionally exceeded 0.003 mg/L (Bow River Water Quality Task Force 1991).

Comprehensive studies determined the effect of stormsewers, combined sewers, municipal andindustrial effluents on water quality in the North Saskatchewan River (Alberta). Stormsewersand combined sewers did not seem to be a significant source of copper during storms: flow-weighted concentrations of total copper were mostly at or below detection limit (Mitchell 1994). Median total copper concentrations of the two municipal wastewater treatment facilities were0.010 and 0.005 mg/L. Median total copper concentrations in the industrial effluents rangedfrom 0.006 to 0.093 mg/L; median concentrations in most effluents were less than 0.020 mg/L(Golder Associates 1994). The highest median concentration was in Sherritt Inc's effluent: theonly company located in Alberta that reported copper releases to NPRI (Section 1.4).

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2 ENVIRONMENTAL FATE AND BEHAVIOUR

2.1 Oxidation states

Copper is the first element of Group 1B of the periodic table and displays four oxidation states:Cu(0), Cu(I), Cu(II), and Cu(III).

Cupric ion (Cu(II) or Cu2+) is the most important oxidation state and is the oxidation stategenerally encountered in water. Cu(II) ions coordinate with six water molecules: thearrangement is distorted in that four molecules closely bound to copper in a planar array and theother more loosely bound in the polar position. Addition of ligands will successively displaceonly the four planar water molecules (Spear and Pierce 1979; U.S. Department of Health &Human Services 1990). Irradiation with UV or reaction with hydrogen peroxide leads to theformation of Cu(I) from Cu(II) (Balzani and Carassiti 1970; Moffett and Zika 1987). However,Cu(I) disappears in milliseconds in oxic environments (Glazewski and Morrison 1995). Inseawater, Cu(II) reduction increases with an increase in chloride concentrations and is likelycontrolled by poorly characterized organic chelators (Moffett and Zika 1987). The proportion ofCu(I) relative to Cu(II) increases with an increase in bicarbonate concentration and an increase inpH. However, Cu(I) is still less than 0.5% of Cu(II) (Stiff 1971).

Cuprous ion or Cu(I) is difficult to study in aquaeous solution because of low solubility (Leckieand Davis 1979) and strong tendency to disproportionate according to the following reaction, inwhich L is a ligand:

(Balzani and Carassiti 1970).

Cu(III) is strongly oxidizing and only occurs in a few compounds. None of these compounds areindustrially important and environmentally significant (U.S. Department of Health & HumanServices 1990).

2.2 Inorganic Complexation

In freshwater free of organic complexing agents, the solubility of Cu(II) is controlled bymalachite (Cu2(OH)2CO3) below a pH of 7 and by tenorite (CuO) above a pH of 7 (Stumm andMorgan 1981). Azurite (Cu3(OH)2(CO3)2) may form instead of malachite depending on thecarbonate and copper concentrations (Mancy and Allen 1977). The main inorganic cupric ion

9

species present in freshwater vary with pH: with increasing pH, the dominant species changefrom Cu2+, CuCO3, Cu(CO3)2

2-, Cu(OH)3- to ultimately Cu(OH)4

2- (Stumm and Morgan 1981).The relative proportion of Cu2+, copperhydroxy and carbonate complexes also depend onalkalinity and the magnitude of stability constants for the formation of complexes. Although pHand alkalinity vary substantially in freshwater systems, Cu2+, Cu(OH)+, Cu(OH)2, and Cu(CO3)2

2-

make up 98% of dissolved inorganic copper. Other chemical species complexed withhydroxides, carbonates, chlorides, sulphate, ammonium, and phosphates make up less than 2% ofdissolved inorganic copper (Nelson et al. 1986). Insoluble coppersulfide forms in the presence ofsulfides (Spear and Pierce 1979; Stumm and Morgan 1981).

Cupric ion is the main toxic species. However, copper species other than Cu2+ are toxic to thewaterflea Daphnia (Borgmann and Charlton 1984). The inorganic species most toxic to cutthroattrout are Cu2+, Cu(OH)+ and Cu(OH)2 (Chakoumakis et al. 1979). However, the magnitude ofthe stability constant of Cu(OH)2 formation is very large, indicating that this copper species israre (Nelson et al. 1986). CuCO3 is generally not considered to be toxic (Chakoumakis et al.1979; Nelson et al. 1986).

2.3 Organic Complexation

Many studies indicate that copper in water is held in solution mainly by complexation withnaturally occurring organic ligands (Mancy and Allen 1977; Spear and Pierce 1979). Infreshwater, organic ligands are more important in binding copper than inorganic ligandscontaining sulfur, phosphorus, chloride, nitrogen. These latter complexes are more important inseawater because of higher concentrations of these ligands (Flemming and Trevors 1989). Dissolved organic-copper include complexes with amino acids, carboxylic acids, humic acids,and various copper chelators (Nelson et al. 1986). Substantial organic-copper complexation alsomay occur in water with relatively low organic content (Nelson et al. 1986). The complexationreaction with fulvic acids and dissolved organic matter is fast (Gardner and Ravenscroft 1991;Lin et al. 1994). The affinity of copper varies among organic compounds: the affinity of copperfor humic acids is greater than for fulvic acids (Huang and Yang 1995). In addition, coppercomplexation capacity increased with decreasing pH and increasing salinity (Gardner andRavenscroft 1991).

The effect on copper toxicity was tested on various organisms with several organic compounds(NTA, EDTA, fulvic acid, amino acids, extracellular polypeptides, algal exudates, humic acid,sewage, glycine, chelators in pulp mill effluent, and TRIS). Chelation complexes are expected tobe relatively non-toxic (Spear and Pierce 1979). However, some of the copper-organiccomplexes were still toxic (Daly et al. 1990b; Borgmann and Ralph 1983). Their toxicity variedwith alkalinity, pH and hardness (Straus and Tucker 1993).

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The presence of organic chelators generally decreased the toxicity of copper to aquaticorganisms. McCrady and Chapman (1979) suggested that EDTA reduced the activity(concentration) of the toxic Cu2+ species. Only two studies indicated that organic chelators didnot ameliorate toxicity: NTA did not affect copper toxicity to algae likely because chelators werepresent in the growth medium (Laube et al. 1980), and fulvic acid did not reduce copper toxicityto the green alga Chlamydomonas whereas humic acid did (Garvey et al. 1991). A more recentstudy on the clawed toad (Xenopus laevis) indicated that fulvic acid only reduced toxicity ifcopper concentrations were less than its pH-dependent solubility limit. At copper concentrationsgreater than the pH-dependent solubility limit, copper toxicity increased with greater fulvic acidconcentrations (Buchwalter et al. 1996).

Copper toxicity was reduced by organic chelators for:

bacteria Pseudomonas Menkissoglu and Lindow 1991; Azenha et al. 1995 algae Anabaena Fogg and Westlake 1953

photosynthesis Steeman-Nielsen and Bruun-Laursen 1976 Chlorella Morrison and Florence 1989; Gächter et al. 1973filamentous algae Shuttleworth and Unz 1991desmids Ivorra 1995Nostoc Mishra et al. 1993Chlamydomonas Garvey et al. 1991; Xue and Sigg 1990Anacystis Lee et al. 1993

plants Elodea Brown and Rattigan 1979invertebrates Simocephalus Giesy et al. 1983

Brachionus Porta and Ronco 1993 copepods Borgmann and Ralph 1984 Daphnia Rao 1985Daphnia pulex Winner 1985Daphnia magna Khangarot et al. 1987; Oikari et al. 1992; Winner

1984 Paratya Daly et al. 1990a

fish guppies Chynoweth et al. 1976 Atlantic salmon Grande 1967; Wilson 1972; Carson and Carson

1972; Zitko et al. 1973 channel catfish Straus and Tucker 1993 fathead minnow Nelson et al. 1986; Erickson et al. 1996rainbow trout Brown et al. 1974

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2.4 Adsorption

Adsorption of copper to colloids and particulates occurs and can constitute a major proportion ofcopper. Compounds such as hydrous metal oxides, organic compounds of high molecularweight, particulates such as clays on organic materials may adsorb copper efficiently (Spear andPierce 1979). Most copper in water was adsorbed and the amount adsorbed increased with anincrease in suspended matter (Mouvet and Bourg 1983). At pH levels above the isoelectric point(i.e., the pH at which the surface area has a zero charge, usually pH>6), the adsorptive surfacebecomes negatively charged, and copper may be adsorbed (Spear and Pierce 1979; Huang andYang 1995). Adsorption of copper to suspended clays and oxides increased with increasing pH(Al-Sabri et al. 1993; Nelson et al. 1986; Davis and Leckie 1978). At pH below 6, desorption ofcopper can occur (O'Connor and Kester 1975 referenced by Spear and Pierce 1979) possibly dueto protons competing with copper for adsorption on the binding sites (Huang and Yang 1995). The presence of organic ligands may increase or decrease adsorption of copper according toDavis and Leckie (1978). Increases in organic content however increased adsorption of copperonto clays and oxides (Nelson et al. 1986).

Adsorbed copper is considered to be of low potency (Spear and Pierce 1979). The addition ofclay reduced copper toxicity (increased the concentration lethal to 50% of the test organisms orLC50) to fathead minnow expressed as total copper (Nelson et al. 1986). The addition of soildecreased copper toxicity to Eurasian milfoil (Myriophyllum spicatum) (Stanley 1974). Ironoxyhydroxides adsorb copper and reduce uptake of copper by molluscs (Tessier et al. 1984).

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3 ENVIRONMENTAL EFFECTS OF COPPER

3.1 Effects of Copper on Fish

Physiological Effects

Copper affected blood characteristics in fish. Rainbow trout exposed to 2 mg/L copper hadhigher blood sugar and plasma enzymes (Nemcsok and Hughes 1988). Erythrocyte counts andhaemoglobin generally increased whereas leucocytes and lymphocytes decreased in brook trout,rainbow trout, and carp (McKim et al. 1970; Dick and Dixon 1985; Svobodova et al. 1994). Mozabique tilapia exposed to copper however showed signs of haemodilution: lowerhaemoglobin levels, similar haematocrit levels, and erythrocytes were swollen compared tounexposed fish (Cyriac et al. 1989). Copper exposure of 0.05 mg/L significantly decreasedhaemoglobin and haemocrit values in Asian catfish (Saccobranchus fossilis) (Khangarot et al.1988). Lower red blood counts, white blood cells, packed cell volume and a gradual decline inoxygen carrying capacity resulted from a sublethal copper exposure of Asian catfish. This mayculminate in respiratory failure (Khangarot and Tripathi 1991). Copper exposure damaged thesurface of erythrocytes (Ahmad and Munshi 1992). The plasma volume in striped bass exposedto copper increased (Courtois and Meyerhoff 1975). Acute copper exposure of carp resulted inan increase in plasma glucose (Svobodova et al. 1994). Brook trout exposed to copper had morePGOT enzyme and total protein content, but had less plasma chloride and osmolarity decreased. After 9 month exposure, only PGOT was decreased (McKim et al. 1970).

Hemorrhaging in cardiac region and base of fins, extruding scales, and darkening of bodycoloration was observed in zebrafish exposed to copper (Weinstein 1978). Copper exposure ofcarp resulted in: production of skin and gill mucus, gills turned grey-red with heavy bleeding, andliver blood vessels were prominent in the body cavity (Svobodova et al. 1994). However, copperexposure of rainbow trout did not result in excess mucus or colour loss (Sellers et al. 1975).

Copper exposure of rainbow trout resulted in respiratory toxicity: heart rate, arterial bloodpressure and lactate concentrations increased, and oxygen tension decreased (Wilson and Taylor1993). Oxygen consumption in bluegills increased initially when exposed to copper, fell belowbase rate and recovered to the base rate after 7 days (O'Hara 1971). Sublethal copper exposure ofbluegills however did not change whole body oxygen consumption after a 9-day exposure, butdecreased oxygen consumption after a 32-day exposure (Felts and Heath 1984). When bluegillswere exposed to increased temperature, whole body oxygen consumption increased; a partialrecovery of oxygen consumption in control fish did not occur in copper-exposed fish (Felts andHeath 1984). Copper affected short-term oxygen consumption in common carp and decreasedammonia excretion. Critical dissolved oxygen levels for the common carp increased from 1.4

13

mg/L to 3.9 mg/L in the presence of 0.022 mg/L copper; no recovery occurred at 0.053 mg/L copper (De Boeck et al. 1995). Fathead minnows exposed to higher copper concentrations wereless tolerant of low dissolved oxygen levels. However, the effect on low dissolved oxygentolerance was not persistent over longer exposure periods (Bennett et al. 1995). No significantchange in oxidative capacity in the gills of rainbow trout was observed (Bilinski and Jones 1973). Also copper did not appreciably alter arterial oxygen tension in rainbow trout (Sellers et al.1975).

Copper exposure of rainbow trout resulted in ionoregulation interference (Wilson and Taylor1993). Copper had little effect on calcium transport across the gills of rainbow trout, butinhibited sodium influx and stimulated sodium efflux (Lauren and McDonald 1986; Reid andMcDonald 1988). High alkalinity reduced the effects of copper on sodium fluxes and on thepermeability of gill membranes. High hardness however did not reduce sodium fluxes or thepermeability of gill membranes (Lauren and McDonald 1986). Exposure of tilapia to copper hadlittle effect on whole body sodium efflux and calcium transport across gill membranes, butinhibited whole body sodium influx (Pelgrom et al. 1995). Copper reduced calcium, sodium andpotassium uptake by brown trout (Sayer et al. 1989).

Copper exposure of rainbow trout changed the histology of the olfactory organ: increase in gobletcells, more mucous cells, vacuoles in sensory epithelium, lesions in sensory and non-sensoryepithelia compared to unexposed fish. Some restoration occurred after fish were removed fromthe copper solution (Saucier et al. 1991).

Cortisol and corticosteroid levels were higher in copper-exposed coho salmon and sockeyesalmon, respectively (Schreck and Lorz 1978; Donaldson and Dye 1975).

Copper affected the immune response of blue gourami to viral and bacterial antigens (Roales andPerlmutter 1977). The immune response of rainbow trout to fetal calf serum (measured asnumber of antibody producing cells) was inversely correlated with copper concentrations(Anderson et al. 1989). The antibody levels against Vibrio were lower in copper-exposed cohosalmon than in controls (Stevens 1977). Asian catfish immunized with sheep red blood cells andexposed to copper had lower antibody titers, reduced number of splenic and kidney plaque-forming cells, phagocytic acitivity in kidney and spleen cells and delayed eye-allograft rejectioncompared to control fish (Khangarot and Tripathi 1991). However, copper did not affect theimmune response of rainbow trout to human red blood cell antigen (Viale and Calamari 1984).

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Behaviour

Copper reduced feeding in several fish species: rainbow trout (Lett et al. 1976; Waiwood andBeamish 1978), Atlantic salmon fingerlings (Grande 1967), and bluegills (Sandheinrich andAtchison 1989). Copper also increased prey handling time of bluegills (Sandheinrich andAtchison 1989) and reduced swimming performance of brown trout (Beamont et al. 1995). Thislack of feeding could have resulted in poor growth.

Copper exposure also resulted in respiratory stress, which affected behaviour of several fishspecies. Guppies exposed to copper showed increased rate and depth of opercular movement,gulping behaviour, and increased swimming activity (Anderson and Weber 1975). Rapid anderratic ventilation was observed in zebrafish exposed to copper. In contrast to guppies, zebrafishwere generally sluggish, with slow or no response to visual or tactile stimuli, and lost equilibrium(Weinstein 1978). Indian catfish were also lethargic, but surfaced frequently to gulp air (Singhand Reddy 1990). Rainbow trout exposed to copper did not appreciably increase coughing(Sellers et al. 1975); signs of stress however were deeper (but not faster) breathing, cramps andataxia (Liepolt and Weber 1958). Copper exposure resulted in signs of restlessness and erraticswimming of the Indian warmwater fish Mystus vittatus and Colisa fasciatus (Pande and Shukla1992).

Avoidance behaviour of copper by fish was summarized by Giattina and Garton (1983). Fishavoided copper concentrations varying from 0.0001 to 0.07 mg/L. Preference or avoidance wasinfluenced by temperature, steepness of the copper gradient. Fish appeared to avoid lowconcentrations but preferred higher concentrations. Two hypotheses were proposed to explainthis ambivalent behaviour: (1) the attraction to high concentrations is a pseudo-attractionresponse caused by chemically induced narcosis, and/or (2) changes in sensitivity of thechemoreceptors at different concentrations may cause initial avoidance with subsequentattraction (Giattina and Garton 1983). Rainbow trout were initially attracted to coppercontaminated water; the attraction was greatest in test with highest copper concentrations. Despite avoidance of copper contamination after the initial attraction, rainbow trout experiencedsignificant mortality (Pedder and Maly 1985). Brown trout avoided simulated Clark Fork Riverwater with a mixture of copper, cadmium, lead and zinc at concentrations half, equal and greaterthan those present in the Clark Fork River (Woodward et al. 1995).

Bioconcentration

Uptake of copper from water by fish is rapid. In rainbow trout, copper was quickly transferred toblood plasma (Zia and McDonald 1994). Liver and gills were the organs accumulating the mostcopper in rainbow trout (Handy 1993), brown bullhead (Brungs et al. 1973) and carp (Svobodovaet al. 1994). Increased copper was also observed in muscles of rainbow trout (Handy 1993).

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Uptake of copper by pumpkinseed sunfish and rainbow trout varied with fish size: copper uptakewas less in bigger fish than in smaller fish (Anderson and Spear 1980a,b).

Depuration

Clearance of copper from pumpkinseed sunfish was fast: halftime was 1.6 to 4.8 hour, and wasfaster in bigger fish (Anderson and Spear 1980a). Copper was cleared from gills to less than50% within 10 hour; clearance of copper from liver took longer than 16 hours (Handy 1992). Asimilar trend was observed in stone loaches: copper loss from gills was fast whereas the liverappeared to retain copper (Solbe and Cooper 1976). Elimination of copper from carp was fastafter fish were removed from copper solution to a water/EDTA mixture (Muramoto 1983).

Acute Toxicity

Early lifestages of brook trout and chinook salmon were more susceptible to copper than olderlifestages (McKim and Benoit 1971; Chapman 1978). Fish size also seemed to affect coppertoxicity. Smaller fish were more sensitive to copper than larger fish for the following species:cutthroat trout (Chakoumakis et al. 1979), rainbow trout (Howarth and Sprague 1978),pumpkinseeds (Anderson and Spear 1980b), and guppies (Spear and Anderson 1975; Andersonand Weber 1975). However, the order of death in young Atlantic salmon was not related to size(Sprague 1964b) nor did size influence copper toxicity to rainbow trout (Anderson and Spear1980b).

Pre-exposure to copper reduced toxicity of copper to rainbow trout. However, this coppertolerance was lost within 7 days (Dixon and Sprague 1981). Intermittent exposure of copper wasmore toxic to steelhead trout than continuous exposure (Seim et al. 1984). Copper toxicity alsoincreased with an increase in temperature (goldfish, channel catfish and trout; Smith and Heath1979) and a decrease in dissolved oxygen (rainbow trout; Lloyd 1961a). Copper acted faster onrainbow trout exposed to higher temperature (Liepolt and Weber 1958). Copper was more toxicto fathead minnow at 12 and 22 oC than at 5 and 32 oC (Richards and Beitinger 1995). Coppertoxicity decreased with an increase in suspended solids (rainbow trout; Brown et al. 1974) andH2S levels (Oseid and Smith 1972).

Copper in water with higher hardness was generally less toxic to fish than water with lowerhardness. Hardness reduced copper toxicity to rainbow trout (Liepolt and Weber 1958; Lloyd1961b), fathead minnow (Nelson et al. 1986; Pickering and Lazorchak 1995; Erickson et al.1996), carp (Peres and Pihan 1991a), channel catfish (Straus and Tucker 1993), and chinooksalmon (Chapman and McCrady 1977). Increased calcium levels had the same effect in reducingcopper toxicity to steelhead trout (Cusimano et al. 1986) and fathead minnows (Nelson et al.

16

1986). However, hardness did not significantly affect survival of juvenile catfish (Wurts andPerschbacher 1994).

Increased alkalinity also reduced copper toxicity to channel catfish (Straus and Tucker 1993) andchinook salmon (Chapman and McCrady 1977). However, increases in alkalinity did notsignificantly affect copper toxicity to fathead minnows (Nelson et al. 1986; Erickson et al. 1996).

The effect of pH on copper toxicity is not simple. If a test species is sensitive to pH, highertoxicity at low pH would result. If species is not sensitive to pH, toxicity may be greater athigher pH due to diminished competition of Cu2+ and H+ ions at receptor sites compared tocompetition at lower pH (Cusimano et al. 1986; Hutchinson and Sprague 1989). In addition, thespeciation of copper changes at different pH (see Section 2.2), which could affect the resultingtoxicity. Copper is more toxic to fathead minnows at lower pH (Nelson et al. 1986; Welsh et al.1993; Erickson et al. 1996). Copper also acted faster in rainbow trout exposed to lower pH (Liepolt and Weber 1958). The toxicity of chelated copper to channel catfish increased at lowerpH (Straus and Tucker 1993). However, mucus secretion increased at low pH which maychange copper species to less toxic forms.

Chronic Toxicity

The chronic toxicity of copper varied among fish species. Hatchability of bluntnose minnoweggs was not affected by copper (Horning and Neiheisel 1979). Survival of common carphatchlings of common carp was affected at copper concentrations greater than 1.0 mg/L copper(Kaur and Virk 1980). Copper delayed hatching of pink salmon but not of sockeye salmon. Initially, the length of sockeye salmon and pink salmon alevins was less at 0.018 and 0.006 mg/Lcopper compared to controls, respectively. However, the difference in body length becameinsignificant when yolk was absorbed (Servizi and Martens 1978). Copper retarded growth anddevelopment of early life stages of brown trout (Sayer et al. 1991). Exposure to copper reducedgrowth of rainbow trout (Lett et al. 1976), Asian catfish (Khangarot and Tripathi 1991), and theIndian warmwater fish species Mystus vittatus and Colisa fasciatus (Pande and Shukla 1992).

Copper exposure in freshwater adversely affected seawater survival. Coho salmon adaptation toseawater was affected at 0.018 mg/L (Stevens 1977). Survival of coho salmon in seawater wasreduced (Lorz and McPherson 1976; Schreck and Lorz 1978). Migration of salmon was alsoreduced when coho salmon exposed to copper in freshwater in the laboratory was released in anearby creek (Lorz and McPherson 1976).

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Mixtures

The joint action in mixtures of two toxicants can be described as:

similar or dissimilar depending on whether the sites of primary action of the twotoxicants are the same or different; and

interactive or non-interactive depending on whether one toxicant does or does notinfluence the biological action of the other toxicant(Alabaster et al. 1994).

Joint action is generally descibed in terms of additive, more than additive (supra-addition orsynergistic) and less than additive. When joint action of two toxicants is additive, each chemicalin the mixture contributes its proportional toxicity. Proportional toxicity is the concentrationpresent in a mixture divided by its toxicity as a single toxicant (e.g., concentration lethal to 50%of the test organisms or LC50 ): [Cu]/LC50. Toxicity in a mixture is more than additive if thetoxicity in the mixture is greater than the sum of the proportional toxicity of the individualchemicals. Toxicity in a mixture is less than additive if the toxicity in the mixture is less than thesum of the proportional toxicity of the individual chemicals.

Mixtures of copper and zinc had different effects on different fish. Acute toxicity of copper andzinc was additive to rainbow trout (Brown and Dalton 1970) and bluegill sunfish (Thompson etal. 1980), was more than additive to young Atlantic salmon (Sprague 1964a; Sprague andRamsay 1965), and was less than additive to juvenile chinook salmon (Finlayson and Verrue1982). Acute toxicity of copper and phenol, copper with zinc and nickel, and copper with zincand phenol mixtures was additive to rainbow trout (Brown and Dalton 1970). Acute toxicity inmixtures of copper and cadmium was additive to juvenile chinook salmon (Finlayson and Verrue1982). Chronic toxicity of copper-cadmium-zinc mixtures was not additive to fathead minnow;however, acute toxicity indicated some interaction (Eaton 1973).

3.2 Effects of Copper on Invertebrates

Physiological Effects

Copper modified the effect of neurotransmittors and ionic current in the simple nerve systems ofthe pond snail Lymnaea stagnalis (S.-Rozsa and Salanki 1990). Exposure of the snailBiomphalaria glabrata to copper likely affected the osmoregulatory physiology: either bysecretion of mucus resulting in suffocation, disruption of membrane permeability, or disruptionof normal cytoplasm function (Sullivan and Cheng 1975). The osmotic and ionic regulation inthe crayfish Orconectes rusticus was also affected: the antennal glands degenerated to varyingdegrees when exposed to low concentrations of copper. Respiratory enzymes were inhibited at

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copper concentrations greater than 1 mg/L (Hubschman 1967a). Filtration by the zebra musselDreissenia polymorpha was inhibited at 0.05 mg/L copper (Mersch et al. 1993). Sublethal levelsof copper (0.25-1 mg/L) resulted in increased respiration rates in the freshwater musselLamellidens marginalis compared to controls (Raj and Hameed 1991).

Bioconcentration

In crayfish, copper concentrations were highest in the hepatopancreas and gills (Cambarusbartoni: Zia and Alikhan 1989, and Alikhan et al. 1990; Procambarus clarkii and P. acutusacutus: Finerty et al. 1990). Highest copper concentrations were in gills and mantle of thefreshwater mollusc Elliptio complanata (Tessier et al. 1984).

Uptake of copper by the clam Anadonta cygnea was linear: depuration of copper from gills didnot occur whereas copper concentrations in foot and mantle decreased in a linear fashion(Salanki and V.-Balogh 1989). Bioconcentration of copper by daphnids was reduced in water ofgreater hardness and higher concentration of humic acids, the latter only in older daphnids(Winner 1985). Uptake of copper by the isopod Asellus aquaticus increased at greatertemperatures, but pH over a range from 5 to 8 did not significantly affect copper concentrationsin the isopod. Elimination of copper from the isopod was not generally observed; copper wasbound in a stable body compartment (Van Hattum et al. 1993).

Copper concentrations in chironomids were positively related to easily reducible copper sedimentconcentrations and negatively with easily reducible iron sediment concentration (Young andHarvey 1991). Copper concentrations in the freshwater mollusc E. complanata were less whenmore amorphous iron oxyhydroxides were present (Tessier et al. 1984). Conversely, copperconcentrations in crayfish (P. clarkii and P. acutus acutus) were unrelated to sediment copperconcentrations (Finerty et al. 1990). Survival of oligochaete worm Lumbriculus was not affectedby copper in sediments (West et al. 1993).

The effect of animal size on the copper concentration in an animal is not clear. Copperconcentrations in the Pacific oyster (Crassostrea gigas) increased with size (Ayling 1974). Copper concentrations were independent of the size of the amphipod Hyalella azteca. Uptake ofcopper was rapid but these amphipods were able to control body burdens gradually and after longexposure periods (Borgmann and Norwood 1995). Metal concentrations (including copper) infour species of mayfly larvae decreased with an increase in size (Jop 1991).

Trichoptera larvae (Plectrocnema conspersa) from a contaminated site (copper concentration inwater 0.84 mg/L) had high concentrations of copper in granules in malpighian tubule cells and

19

subcuticular regions. Darlington and Gower (1990) suggested that these pigment granulesprevent toxic effects.

Behaviour

Copper reduced the activity of many mollusks. Feeding of the snail Campeloma decisum onclam meat ceased at copper concentration of 0.015 and 0.028 mg/L; the opercululum closed athigh copper concentrations (Arthur and Leonard 1970). Exposure of A. cygnea to 0.01 and 0.1mg/L copper resulted in less active clams (Salanki and V.-Balogh 1989). At high copper dosesthe snail Biomphalaria glabrata retreated in its shell (Sullivan and Cheng 1975). Thefreshwater snail Thiara tuberculata reduced locomotion when exposed to 5 mg/L copper bywithdrawing inside its shell. Exposure to 1 mg/L copper resulted in normal movement for thefirst 10 days and a gradual reduction up to 20 days; oxygen consumption was severely reduced(Mule and Lomte 1994).

Exposure of tubificid worms (Tubifex tubifex) to copper resulted in sharp twisting motions,segmentation and disintegration (Brkovic-Popovic and Popovic 1977). Copper concentration of0.125 to 0.5 mg/L resulted in clumping of the oligochaete worm Lumbriculus variegatus, higherexposures caused the worms to break up in relatively large pieces (Baily and Liu 1980).

Acute Toxicity

No single life stage was consistently most sensitive to copper. Eggs of the snail Amnicola wereless sensitive than adults (Rehwoldt et al. 1973), whereas crayfish eggs (Orconectes rusticus)were most sensitive (Hubschman 1967b). The first instars of the midge larvae Chironomustentans and Polypedilum nubifer were the life stage most sensitive to copper (Nebeker et al.1984b; Hatekayama 1988). Toxicity of copper to the amphipod Hyalella azteca was consistentacross all age classes (Collyard et al. 1994). Size of the freshwater shrimp Gammarus pulex didnot affect toxicity to copper (Stephenson 1983).

A number of environmental factors affected copper sensitivity of invertebrates: water hardness,alkalinity, ionic strength, and pH.

An increase in hardness or alkalinity decreased the toxicity of copper to tubificid worms(Brkovic-Popovic and Popovic 1977). An increase in water hardness also reduced the acutetoxicity to the cladoceran Ceriodaphnia dubia (Belanger et al. 1989; Belanger and Cherry 1990);the rotifer Philodina acutiformis (Buikema et al. 1974); snails Stagnicola and Physa sp. (Howardet al. 1964); freshwater shrimp Gammarus pulex (Stephenson 1983). Copper was more toxic to

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the Australian freshwater shrimp Paratya australiensis at lower ionic strength either due toincreased uptake (gill/respiration) or due to combined copper and ionic stress (Daly et al. 1990b).

Alkaline stress (pH 10.5) decreased toxicity of copper to the snail Goniobasis livescens (Paulsonet al. 1983). An increase in pH reduced copper toxicity to the waterflea Moina irrasa. Anincrease in water temperature resulted in faster death of the fleas when exposed to copper (Zouand Bu 1994). An increase in pH reduced acute toxicity of copper to the cladoceranCeriodaphnia dubia, but did not affect chronic toxicity (Belanger and Cherry 1990).

Survival of the waterflea Daphnia magna was greater when food was added to the copper testsolutions (Biesinger and Christensen 1972).

Chronic toxicity

Copper interrupted the lifecycle of a caddisfly and a clam. Copper concentrations equal to orgreater than 0.017 mg/L prevented completion of the lifecycle of the caddisfly Clistoroniamagnifica. At 0.017 mg/L copper, wings were either crumpled, incompletely inflated orflattened, legs were malformed, detachment of pupal skin was incomplete and mating did notoccur (Nebeker et al. 1984a). The early life stage of the clam Anadonta cygnea was moresensitive to copper than adults. Glochidia could not reopen and were therefore unable to attachto fish and continue their development (Huebner and Pynnonen 1992).

Copper also affected growth. Sublethal levels of copper (0.25-1 mg/L) resulted in greater weightloss of the freshwater mussel Lamellidens marginalis compared to controls (Raj and Hameed1991).

Mixtures

The waterflea Daphnia was exposed to an equitoxic mixture of As, Cd, Cr, Cu, Hg, Ni, Pb, Zn:chronic effects were nearly additive (Enserink et al. 1991). Zebra mussels (D. polymorpha) wereexposed to metal mixtures. The toxic effect on feeding (filtration rate) of copper and zinc wereless than additive, the effect of copper and cadmium more than additive, and the effect of thecopper-cadmium-zinc mixture additive (Kraak et al. 1994). During prolonged exposure of zebramussels, the effect of copper, zinc and cadmium mixtures on filtration rates was no longeradditive (Kraak et al. 1993).

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3.3 Effects of Copper on Amphibians

The toxic effects of copper on amphibians are summarized in Appendix 1. In a study of severalamphibian species, embryos were generally more sensitive to copper than larvae (Birge andBlack 1979). Larval LC50s for 2 toad (Fowler's toad Bufo fowleri and narrow-mouthed toadGastrophryne carolinensis) and two frog (northern leopard frog Rana pipiens and southern graytreefrog Hyla chrysoscelis) species were not significantly changed from LC50s for egg hatching(Birge and Black 1979). However, larvae of marbled salamanders (Ambystoma opacum) weremore sensitive to copper than embryos: the LC50 was lower when the toxicity study was extendedbeyond hatching. Metamorphosis of boreal toad (Bufo boreas) larvae was not affected by copper(Porter and Hakanson 1976). Copper reduced growth of frog (Rana pipiens) tadpoles (Lande andGuttman 1973).

Behaviour was also affected by copper. At higher copper concentration, toad tadpoles (Bufomelanostictus) surfaced more, had erratic body movements, and loss of equilibrium (Khangarotand Ray 1987a). In frogs (Rana pipiens) exposed to copper above 0.0015%, mucus outputincreased, heart rate initially increased than decreased, neuromuscular coordination wasdisturbed, and general activity was lower (Kaplan and Yoh 1961). Similar effects were observedwith clawed toad (Xenopis laevis) larvae (Fingal and Kaplan 1963). Frog tadpoles (Microhylaornata) also showed signs of irritability when exposed to copper, followed by loss of equilibriumand death (Rao and Madhyastha 1987). American toad (Bufo americanus) tadpoles avoidedconcentrations of 0.1 mg/L copper but were attracted to 0.93 mg/L (Birge et al. 1993).

3.4 Effects of Copper on Plants

Exposure to copper reduced growth of plants, photosynthesis, respiration, and nitrogen fixation,and resulted in physiological damage and deflagellation.

Growth of blue-green and green algae was delayed when algae were exposed to copper (Bartlettet al. 1974; Foster 1977; Rosko and Rachlin 1977; Laube et al. 1980). Copper exposure delayedgrowth of the green alga Scenedesmus, reduced oxygen devolution, destroyed chlorophyll-a, andresulted in severe membrane decomposition, peroxidation of lipids (Sandmann and Boger 1980). Copper exposure resulted in a reduction in the green alga Chlorella vulgaris growth and anincrease in cell size (Rosko and Rachlin 1977), up to 20 times the normal volume (Foster 1977). The cell size of Scenedesmus quadricauda was also enlarged after copper exposure (Khobot'yevet al. 1975). However, copper had little effect on average cell volume of the green algaeSelenastrum capricornutum or Chlorella stigmatophora (Christensen and Sherfig 1979). Deflagellation of the green alga Chlamydomonas was a more sensitive indicator of coppertoxicity than population growth or encystment (Garvey et al. 1991).

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The blue-green alga Anabaena took up more copper than the green alga Scenedesmus and wasmore sensitive to copper. Anabaena did not recover after re-inoculation at copper concentrationsof 0.25 mg/L, whereas Scenedesmus recovered at concentrations less than 0.8 mg/L (Gibson1972). Growth rate was inversely proportional to copper levels taken up by two strains of thegreen alga Chlorella vulgaris. Exclusion of copper was indicated as a mechanism to explain thedifference in copper tolerance between the two strains of Chlorella vulgaris (Foster 1977).

Effects of copper on photosynthesis of the green alga Chlorella were fast (<24 hr). Copper wasmore toxic at higher light intensity (Steemann Nielsen et al. 1969). Cell division by Chlorelladuring dark periods was inhibited at lower copper concentrations than cell division during lightperiods (Kanazawa and Kanazawa 1969). Copper reduced photoreduction of NADP+

(photosystem I - PSI) and DCIP (Photosystem II - PSII) in the green alga Ankistrodesmus falcatus(Shioi et al. 1978) and inhibited electron transport from watersplitting system to primary acceptor(Samson and Popovic 1988). In the bluegreen alga Anacystis nidulans, copper reduced PSIIactivity by causing structural alterations in the chlorophyll-proteins of the PSII complex. Inaddition, copper effected the light harvesting complex (Gupta and Singhal 1995). Copper alsoreduced PSII activity of the green alga Chlorella pyrenoidosa. The PSII inhibition was morepronounced at higher light levels, indicating that copper decreased the tolerance tophotoinhibition (Vavilin et al. 1995).

Copper exposure significantly reduced chlorophyll-a concentrations, photosynthesis and nitrogenfixation. Bluegreen algae were most affected (Elder and Horne 1978). The copper concentrationinhibiting growth of the bluegreen alga Cylindrospermum by 50% (IC50) was 0.025 mg/L;toxicity was reduced as pH increased (Khare and Bisen 1991). Copper sulphate caused areduction of nitrogen fixation and physiological damage (with a concomitant release of dissolvedorganic carbon and geosmin) of Aphanizomenon flos-aquae (Peterson et al. 1995). Darkrespiration increased of the waterweed Elodea increased with copper exposure compared tocontrols. However, chelators could overcome this effect (Brown and Rattigan 1979).

Toxicity

Toxicity tests with plants are performed in special growth media. These growth media generallycontain buffers and organic compounds (like EDTA). As explained in Chapter 2, these organiccompounds may change the speciation of copper and result in less toxic forms. Coppercomplexes were less toxic than CuCl2 (Khobot'yev et al. 1975; Morrison and Florence 1989;Stauber and Florence 1989; Lee et al. 1993). Copper may also bind to algal surfaces or to algalexudates resulting in lower toxicity. In a test with the green alga Chlamydomonas, the speciationof copper was altered more by binding to algal exudates than by binding to algal surfaces (Xueand Sigg 1990).

23

The toxic effects of copper on plants are summarized in Appendix 2. Toxicity endpoint forplants ranged from 0.006 mg/L (lowest observed effect concentration affecting photosynthesis-Gächter et al. 1973) to over 25 mg/L copper (growth Brady et al. 1994).

Mixtures

Several mixtures of metals including copper were tested on plants. More than additive effectswere observed with copper and manganese (Christensen and Sherfig 1979), copper and cadmium(Lasheen et al. 1990; Rachlin and Grosso 1993), copper and zinc (French and Evans 1988),cadmium and cobalt (Rachlin and Grosso 1993), whereas less than additive effects wereobserved with copper and lead (Christensen and Sherfig 1979), copper and iron (Jain et al. 1992).

3.5 Effects of Copper on Other Microorganisms

Copper concentration of 1.1 mg/L inhibited respiration 40% by Escherichia coli bacteria(Dorward and Barisas 1984). Respiration by E. coli was reduced at 5 mg/L (Jardim et al. 1990). Addition of copper greater than 1 mg/L to indoor ponds reduced E. coli numbers (Jana andBhattacharya 1988). Oxygen depletion by a bacterial community was reduced at a copperconcentration of 0.5 mg/L (Bauer et al. 1981).

Results from several microbial bioassays were compared: Microtox assay was the most sensitivetest with EC50s (effect concentration that causes an effect in 50% of the test organisms) of 3.8mg/L, 0.1-0.2 mg/L, and 1.2 mg/L copper (Dutka and Kwan 1981; Reteuna et al. 1989; Codina etal. 1993, respectively). However, Microtox test results were variable between differentlaboratories: EC50 values varied from 8 to 20 mg/L copper (Dutka and Kwan 1981).

Treatment of 6 mg/L copper resulted in high mortality of Pseudomonas syringae bacteria(Azenha et al. 1995). Maximum tolerated copper concentration by 17 strains of this species were0.12 to 0.03 mg/L (Menkissoglu and Lindow 1991). Copper exposure of 13 different protozoanspecies resulted in lethal levels from 0.1 to greater than 100 mg/L with a median toxicity limit of1.6 mg/L after 24 hours. The LC50 value for the protozoan Vorticella after a 3-hour exposurewas 1.8 mg/L copper (Ruthven and Cairns 1973). The toxic threshold level for cellmultiplication was 0.03 mg/L for the bacterium Pseudomonas putida and 0.11 mg/L for theprotozoan Entosiphon sulcatum (Bringmann and Kuhn 1980). The ciliate protozoanSpirostomum ambiguum was very sensitive to copper; the LC50 was 0.004 mg/L copper at lowhardness (2.8 mg/L CaCO3) and 0.005 at high hardness (250 mg/L CaCO3) (Nalecz-Jawecki et al.1993). Toxicity effects of copper on the protozoan Paramecium tetraurelia were temperaturedependent; toxicity increased at higher temperature (Szeto and Nyberg 1979). Threshold toxicityof the planarian Polycelis nigra was 0.47 mg/L copper (Jones 1940).

24

Exposure to 24 mg/L copper reduced the number of protozoan species from an initial 46 speciesto 7 species after 24 hour exposure. Partial recovery was observed after 144 hours when 14species were present (Cairns and Dickson 1970). Colonization of artificial "islands" around anepicenter by protozoans was affected by 0.42 mg/L copper (Cairns et al. 1980). Exposure ofmicrobial communities to 0.01 to 0.02 mg/L copper reduced species richness and biomass (Prattet al. 1993).

3.6 Effects of Copper on Microcosms and Ecosystems

Field experiments/studies

Separate sections of Convict Creek, an oligotrophic Sierra Nevada creek in California, weredosed continuously with different CuSO4 concentrations during two 1-year experiments (startingin 1978 and 1979). The number of species of ciliates and rotifers were affected at 0.007 mg/L(LOEC) and 0.005 mg/L during 1978 and 1979, respectively. No observed effect was observedat 0.0025 mg/L for both years. Rapid recolonization by ciliates and rotifers occurred after thecopper additions were halted (Leland and Kent 1981). Copper additions of 0.005 to 0.01 mg/Lduring 1979/80 almost totally inhibited the filamentous green alga Spirogyra, reduced the greenalgae Cladophora and Mougeotia (at 0.01 mg/L), the bluegreen alga Lyngbia, and the diatomAmplipleura. However, periphyton biomass did not decline due to an increase in the diatomAchnanthes (Leland and Carter 1984).

Benthic invertebrate communities in the Clinch River, Virginia, were affected by the effluent of acoal-fired generating plant. Concentrations of copper and zinc were elevated in water below theplant discharge. Fewer taxa and individuals were observed below the plant discharge. Ephemeroptera were most affected: they did not recover at sites further downstream. Tanytarsinichironomids were also highly sensitive. However, Orthocladiini and Hydropsychidae were moretolerant: abundance was greater at downstream recovery stations than at upstream control sites. Benthic commmunities upstream of the plant discharge were established on trays and transportedto experimental streams supplied continuously with New River water. Copper and zinc wasadded (control, 0.012 mg/L copper and zinc, and 0.05 mg/L copper and zinc). Benthicinvertebrate abundance and number of taxa decreased significantly in the experimental streamsexposed to copper and zinc. The resulting benthic invertebrate community was very similar tothe community observed in the Clinch River below the plant discharge (Clements et al. 1988).

Shayler Run (Ohio) was dosed continuously with 0.12 mg/L copper during 75% of the studyperiod (February 1970 to October 1972). Information from December 1967 to 1970 was used asbackground information. CuSO4 additions affected fish species except for orange throat darter. The effects were death, avoidance of high copper concentrations and restricted spawning.

25

Bluntnose minnows and stonerollers moved out of the copper-dosed area. Four of the fiveabundant macroinvertebrate groups were essentially eliminated (scuds, sowbugs, mayflies andriffle beetles). The fifth abundant macroinvertebrate group (chironomids) flourished in thecopper-dosed area (Geckler et al. 1976). The macroinvertebrate community changed withdistance downstream. The number of individuals and number of species were were lower at siteswith higher copper concentrations (Winner et al. 1975). The periphyton community alsochanged: the dominant diatom (Cocconeis placentula) and filamentous alga (Cladophoreglomerata) were replaced by three other species of diatoms (Nitzschia palea, Navicula nigrii andN. seminulum). A blue-green alga and two desmid species were more abundant in the copper-treated reach than in the control reach (Weber and McFarland 1981).

A copper-lead-zinc mine used to discharge metal mining waste into Butte Lake, BritishColumbia. After termination of this discharge, lakewater copper concentrations dropped from0.17 to 0.04 mg/L. The phytoplankton species composition changed, zooplankton communitywas more diverse, and muscle metallothionein levels in rainbow trout returned to backgroundlevels. Hepatic metallothionein levels were reduced in rainbow trout after discharge to the lakewas eliminated. However, metallothionein levels in cutthroat trout and Dolly Varden did notchange after discharge was stopped (Deniseger et al. 1990).

Copper was added to Cazenovia Lake in New York for algacidal purposes. CuSO4 was addedthree times, which would have resulted in a completely mixed copper concentration of 0.051mg/L. The result was a minor temporary decrease in chlorophyll-a levels, subtle changes in algalcomposition, and bacteria were affected but recovered fast. Plants and zooplankton were notaffected by the copper addition (Effler et al. 1980).

Coppersulfate addition to ponds resulting in 3 mg/L copper partially controlled waterweed Elodianutallii and completely controlled the pondweed Potamogeton crispus, green alga Oedogoniumsp and stonewort (Chara). A smaller dose of 1 mg/L copper completely controlled thefilamentous green alga Spirogyra, but none of the other species. Removal of these species wasonly temporary; green and bluegreen mats and scum developed by day 14 to 17. Ostracods andcopepods were not affected, whereas number of cladocera and rotifers were depressed but laterrecovered. Decomposition of these plants decreased dissolved oxygen levels and increasedhydrogensulfide (H2S) concentrations (McIntosh and Kevern 1974; McIntosh 1974).

Copper was added once to several enclosures in Lake Bure, Denmark (Gustavson and Wängberg1995). Copper concentrations were constant after this initial dose. Phytoplankton in theenclosures with copper were affected by copper: 14C dioxide fixation was lower in thoseenclosures with higher copper concentrations. However, the phytoplankton communitiesexposed to copper developed tolerance over time. After 12 days, short-term EC50 values were

26

higher for the copper exposed phytoplankton communities than for the control and lakecommunities. In addition, algal biomass recovered in the copper-exposed enclosures after aninitial decrease.

Laboratory Microcosms

Continuous CuSO4 additions of 0.0093 mg/L copper to a flow-through microcosm changed thestructure of the microcosm from an autotrophic to a heterotrophic system after 32 weeks (Hedtke1984).

An aquatic microcosm was set up with primary producers, protozoa, rotifers, oligochaetes andbacteria. An addition of 0.7 mg/L to a static version of the microcosm eliminated the rotifers,oligochaetes, the green alga Scenedesmus and blue-green algae. Addition up to 1.2 mg/L copperdid not cause severe elimination in a continuous flow system; bacteria and the green algaChlorella increased and blue-green algae, rotifers, oligochaetes disappeared (Sugiura et al. 1982).

Aquatic microcosms with 10 algal species and 5 invertebrate species were subjected to 0, 0.5 and2 mg/L copper. At the 2 mg/L copper, algae and grazers were severely affected. At 0.5 mg/Lcopper, the grazer Daphnia was unable to survive; this allowed accumulation of algal biomassand a rotifer bloom. The algal dominance changed from the bluegreen algae Nitzschia andLyngbia to the green algae Ankistrodesmus and Scenedesmus (Harass and Taub 1985).

A naturally colonized microcosm with protozoans, bacteria, fungi, diatoms, algae, rotifers,annelids, insects and crustaceans was exposed to CuSO4 in a flow-through experiment. After 21days, taxonomic richness was not affected at 0.0066 mg/L (NOEC), but was affected at 0.0127mg/L (LOEC). Other measurements EC5 for chlorophyll-a and ATP were more sensitive thantaxonomic richness; however these measures may not be sensitive to other toxicants and are alsosubject to natural changes to individual species (Pratt et al. 1987).

Predator-prey interaction were evaluated with copper concentrations up to 0.3 mg/L. Copper wasmore acutely toxic to the protozoan predator Didinium nasutum than to its protozoan preyParamecium caudatum. The intrinsic rate of growth of both predator and prey species wasgreater when copper concentrations were between 0.03 and 0.18 mg/L compared to controls. At0.3 mg/L copper, the intrinsic rate of increase of the predator D. nasutum became negative. Theequilibrium density of the prey P. caudatum tended to decrease as copper concentrationsincreased, whereas the equilibrium concentration of the predator D. nasutum did not change. Thisresulted in a destabilization of the predator-prey interaction. At 0.3 mg/L copper, the equilibriumdensity of both predator and prey increased: predator efficiency decreased allowing the prey

27

equilibrium density to increase (Doucet and Maly 1990).

Biomagnification

Trace metal levels in the littoral foodweb of the Maarsseveen Lakes were studied. Copper levelsin predators were low and equal to those in filter feeders, but higher than in sediments. Depositfeeders contained higher copper levels. Organisms containing haemocyanin (crustaceans,gastropods and bivalves contained higher amount of copper. There was no evidence ofbiomagnification as predators contained less copper than their prey (Timmermans et al. 1989). Inthe Butte Lake (British Columbia) system, copper content decreased from zooplankton tostickleback and to cutthroat trout, indicating the lack of biomagnification (Roch et al. 1985). Copper concentrations in water, sediment, and various aquatic biota at several sites in theSacramento River basin indicated no sign of magnification of copper along the foodchain (Saikiet al. 1995).

Copper was studied in the various compartments of the Wanapitei River, southwest of Sudbury(Ontario). Concentration factors (CF) ranged from 149 and 156 for bullhead and pickerel(omnivorous and carniverous fish, respectively) up to 17667 for periphyton. There was noevidence of biomagnification: the CF for predators was lower than the CF for plankton(zooplankton 3444; Hutchinson et al. 1976). Biomagnification was also not evident in theseveral lakes. Concentrations of copper in biota decreased with increasing trophic level(Prahalad and Seenayya 1986; Radwan et al. 1990).

Biomagnification in a microcosm containing algae, protozoa and metazoa was not observed:algae contained 10 to 100 times more copper than the waterflea Daphnia (Jin et al. 1991).

3.7 Genotoxicity of Copper

Copper is not known to cause cancer (U.S. Department of Health and Human Services 1990). No data are available to suggest that copper is a reproductive or teratogenic toxin (MacLarenPlansearch and FDC Consultants 1985). Human data suggest that copper does not produceteratogenic effects (MacLaren Plansearch and FDC Consultants 1985).

Some data on fish suggest that copper is a a chemical that causes structural defects that affect thedevelopment of an organism (teratogen). Treatment of fish eggs with copper however resulted insignificant numbers of teratic larvae. Larval survival of species with a high copper tolerance(bass, goldfish and catfish) was not affected by gross teratogenic impairment. However, Birgeand Black (1979) indicated that the sensitive developmental stages of rainbow trout may be

28

affected as much by teratogenic effects as by egg mortality. Teratogenic effects on rainbow troutincreased at copper concentrations greater than 0.01 mg/L. Skeletal defects (spinal column andjaw) were the most sensitive indicators of teratogenic effects (Birge and Black 1979). Thedevelopment of common carp was also affected by copper. Fifty percent of the hatchlings wereabnormal after exposing eggs to copper concentrations of 3.5 mg/L. Deformed vertebral column,downward curvature of the caudal region and undeveloped body behind the yolk sac were observed in these hatchlings (Kaur and Virk 1980). In a different study, teratogenic effects werealso observed in common carp at much lower copper concentrations: 0.051 mg/L at pH 7.6 and0.019 mg/L at pH 6.3. The effects included a deformed head, spinal column, upper jaw, and asmall or absent swim bladder (Stouthart et al. 1996).

3.8 Metal-binding Proteins (Biomarker)

Metallothioneins (MTs) are low molecular weight proteins with a high content of certain tracemetals. Their characteristic amino acid composition (high cysteine content and poor in aromaticamino acids), sequence and spectroscopic properties make these proteins unique. The highcysteine content gives MT a very high binding capacity for metals. The first compound with allthese features was first detected in equine kidney cortex. Since this original discovery, severalmetalloproteins with similar biochemical proteins have been identified (Hodson 1988; Carpene1993).

The role of MTs in aquatic species is unknown (Hodson 1988; Petering et al. 1990). Potentialroles include: protection against toxicity (through chelation of reaction with MTthioles) ornormal cellular function (regulation of zinc metabolism and a role in copper metabolism). According to Cosson et al. (1991), it is probable that MTs participation in detoxificationmechanisms are due to fortuitous interactions of foreign cations with the normal homeostasismechanism for zinc and perhaps copper. In addition to heavy metal exposure, MT can beinduced by a large number of diverse agents, and by handling or not feeding fish. Seasonalvariations of MT levels also occur. MT measurements include protein with and without boundmetal; the latter may not provide information on the stressor. Freshwater fish contain a largebasal level of MT-like zinc and copper binding proteins. Extensive knowledge from direct studyof non-mammalian organisms is needed to determine whether MT can be used for environmentalmonitoring purposes (Petering et al. 1990). Not only should the identity and role of other metalbinding proteins be understood (Hodson 1988), also the consequence of biochemical changes atthe organism level to higher levels of organization (population and ecosystem level) need to beunderstood (Thomas 1990).

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4 DERIVATION OF ACUTE AND CHRONIC GUIDELINES

4.1 Final Acute Value and Alberta Acute Guideline

Primary acute toxicity data were available for 29 fish species from five different orders(Salmoniformes, Cypriniformes, Perciformes, Siluriformes, Atherinoformes) and 10 differentfamilies (Salmonidae, Cyprinidae, Plotosidae, Percidae, Centrarchidae, Centroponidae,Ictaluridae, Melanotaeniidae, Cyprinidontidae, and Poeciliidae) (Appendix 3). According to theProtocol to Develop Alberta Water Quality Guidelines for Protection of Freshwater Aquatic Life(Alberta Environmental Protection 1996), the minimum fish data requirements for deriving a fullacute copper guideline are therefore amply met.

Primary acute toxicity data were available for 24 invertebrate species from 11 different orders(Isopoda, Decapoda, Diptera, Cladocera, Ephemeroptera, Amphipoda, Trichoptera, Rotatoria,Annelida, Gastropoda, and Plecoptera) (Appendix 4). The minimum "non-fish" datarequirements for deriving a full acute guideline (Alberta Environmental Protection 1996) aretherefore amply met. Only one study provided primary acute data for amphibians: in a 48-hr testof the two-lined salamander (Eurycea bislineata) 1.12 mg/L copper resulted in an 50% mortality(Dobbs et al. 1994, Appendix 1).

Because hardness was found to modify copper toxicity, relationships between acute coppertoxicity and water hardness were investigated. Acute copper toxicity values were available atdifferent water hardness for five fish species: bluegill, chinook salmon, fathead minnow, guppies,and rainbow trout. However, the range of water hardness for the bluegill and guppy data wassmall (35 to 110 mg/L CaCO3 and 67 to 144 mg/L CaCO3, respectively), and the number of datapoints was small (five and four, respectively). These data were therefore not considered inderiving a relationship between water hardness and acute copper toxicity. Acute copper toxicityvalues were also available at different water hardness for three invertebrate species:Ceriodaphnia dubia, Dapnia magna, and Daphnia pulex. However, the range of water hardnessfor the Daphnia magna was small (85 to 143 mg/L CaCO3), and the number of data points wassmall (three). The range of the Daphnia pulex data was large (45 to 230 mg/L CaCO3). However, the number of the acute toxicity values at low hardness varied almost 10-fold and onlyone acute toxicity value was available at high hardness. Therefore, the Daphnia data were notused in deriving a relationship between water hardness and acute copper toxicity.

Regression relationships for the four remaining species (chinook salmon, fathead minnow,rainbow trout, and Ceriodaphnia dubia) were derived. A natural logarithmic transformationimproved the relationships: guidance as outlined in Alberta Environmental Protection (1996) wastherefore followed. Some of the basic information is provided in Table 4: the detailed

30

calculations are provided in Appendix 5. According to the ancova analysis, the pooled slope issignificant and the slopes for the four different species are similar.

Table 4. Statistical information on regression and ancova analyses (Sokal and Rohlf 1981)for derivation of an acute copper guideline.

n r2 P Slope

Chinook salmon 7 0.901 <0.002 0.688565

Fathead minnow 26 0.363 <0.002 0.91816

Rainbow trout 30 0.759 <0.001 1.03995

Ceriodaphnia dubia 17 0.591 <0.01 1.021908

pooled slope (0.979123) is significant: F1,2=138.4879, P<0.01

slopes for species are equal: F3,72=0.616991, P>0.5

With the pooled slope, the available primary acute data were adjusted to a common waterhardness (100 mg/L CaCO3), according to

in which Y is the ln-transformed toxicity value adjusted to the water hardness of 100 mg/LCaCO3 (Z), W is the original acute toxicity value at the original water hardness (X), and V is thepooled slope (0.979123).

The following adjustments in the data from Appendix 2 and 3 were made:• 96-hr LC50 for Hydropsyche betteni was set at 64 mg/L (as opposed to >64)• the total copper LC50s for Paratya australensis and fathead minnow were used• only the LC50s for sensitive lifestages were used: swim-up chinook salmon and steelhead

trout (Chapman 1978), alevin pink salmon (Servizi and Martens 1978), 1.2 gpumpkinseed (Spear and Anderson 1975), fry sockeye salmon (Servizi and Martens 1978)

• the LC50s for adult and juvenile steelhead trout (Chapman and Stevens 1978; Seim et al.1984) were not used as these were not sensitive lifestages

After adjusting the acute toxicity values to a common water hardness of 100 mg/L CaCO3, thespecies mean acute values (SMAV) were calculated as outlined in Alberta Environmental

31

Protection (1996). The SMAVs varied: for fish species from 0.066 mg/L (chinook salmon) to2.294 mg/L (bluegill sunfish); for invertebrate species from 0.011 mg/L (Caridina) to 139.871mg/L (Hydropsyche); for amphibians 1.02 mg/L (2-lined salamander; Appendix 5). The genusmean acute values (GMAV) were calculated as outlined in Alberta Environmental Protection(1996).

The GMAV for Caridina sp. (Williams et al. 1991) was deleted from the calculation of the finalacute value. This datapoint was eliminated because it was from an Australian species that wasmore sensitive than all the North American species. The resulting final acute value (FAV, at 100mg/L CaCO3 water hardness) was 0.0320 mg/L. No commercially, recreationally andecologically important species was more sensitive than this calculated FAV. The Alberta acuteguideline (at 100 mg/L CaCO3 water hardness) is therefore 0.0160 mg/L. Following the AlbertaEnvironmental Protection (1996) guidance for deriving an acute equation, the resulting acutecopper guideline (in mg/L) is:

Figure 1 presents all primary genus mean acute values (adjusted to 100 mg/L CaCO3 waterhardness) in relation to the Alberta acute guideline.

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33

Alberta Chronic Guideline

Final Chronic Value

Primary chronic toxicity data useful for deriving a chronic guideline were available for 11 fishspecies from three different families (Salmonidae, Cyprinidae, Esocidae; Appendix 6) and for 8invertebrate species from 5 different orders (Diptera, Cladocera, Amphipoda, Trichoptera,Pelycopoda; Appendix 7). According to the Protocol to Develop Alberta Water QualityGuidelines for Protection of Freshwater Aquatic Life (Alberta Environmental Protection 1996),the minimum data requirements for deriving a full chronic copper guideline are therefore amplymet. The chronic toxicity data consisted of a no-observed-effect-concentration (NOEC) and alowest-observed-effect-concentration (LOEC). The geometric mean of the NOEC and LOECvalues (MATC) was calculated and used in further analyses.

Again, the relationship between copper toxicity and water hardness was investigated. Chroniccopper toxicity values were available at different water hardness for two fish species and oneinvertebrate species: brook trout, fathead minnow, and Ceriodaphnia dubia. The results of theregression analyses presented in Table 5.

Table 5. Statistical information on regression analyses for a chronic guideline (Sokal andRohlf 1981).

n untransformed data ln-transformed data

r2 P r2 P

Brook trout 4 0.127 >0.64 0.05 >0.77

Fathead minnow 6 0.827 <0.01 0.70 <0.04

Ceriodaphnia dubia 4 0.056 >0.76 0.08 >0.72

The range in water hardness for all threespecies was large. However, the fatheadminnow data resulted in the onlysignificant relationship between waterhardness and chronic copper toxicity(P<0.05). As illustrated in Figure 2,chronic toxicity data are available only forlow and high water hardness and not forany intermediate water hardness. The

34

chronic toxicity data did not indicate a relaionship between copper toxicity and water hardness. Therefore, a water hardness relationship for chronic toxicity data was not derived.

A chronic guideline equation relating chronic toxicity to water hardness can be calculated fromthe acute guideline by applying an acute-to-chronic ratio (ACR) (Alberta EnvironmentalProtection 1996). Many primary studies provide ACR values (Table 6).

Table 6. Primary acute-to-chronic ratios (ACR) from copper toxicity studies.

Species ACR Water Hardness (mg/L CaCO3)

Reference

Rainbow trout 8.182 25 Marr et al. 1995/1996

Sockeye salmon 3.514 83 Servizi and Martens 1978

Bluegill 52.381 45 Benoit 1971

Pink salmon 3.48 83 Servizi and Martens 1978

Bluntnose minnow 5.349 200 Horning and Neiheisel 1979

Fathead minnow 1.9177.075

18.333

20231

204

Pickering et al. 1977Mount and Stephan 1969Geckler et al. 1976

Ceriodaphnia dubia 1.5835.9334.6049.073

3610094

179

Carlson et al. 1986Spehar and Fiandt 1986Belanger et al. 1989Belanger et al. 1989

Chironomus tentans 22.735 36 Nebeker 1984b

Daphnia magna 13 85 Blaylock et al. 1985

The minimum ACR requirement (primary ACR for fish, invertebrate and the acutely sensitiverainbow trout: Alberta Environmental Protection 1996) is therefore met. However, the ACR dataare not conclusive:

• the ACR seems to increase with water hardness for Ceriodaphnia, but not for fatheadminnow,

• ACR varies widely within a species (almost an order of magnitude for fathead minnowwith similar water hardness; Table 6),

• ACR does not seem to increase or decrease with the acute toxicity of a species (Figure 3),• ACR between species varies over a factor of 10 (Table 6).

35

Therefore, an ACR is not used toadjust the acute guideline equationagainst water hardness to a chronicguideline equation.

Because the chronic toxicity data forbrook trout and Ceriodaphnia dubiaindicated no relationship betweenwater hardness and chronic toxicityand an ACR adjustment to the acuteguidelines could not be used, norelationship with water hardness wasdeveloped for a chronic guideline.

The species mean chronic values(SMCV) were therefore not adjusted for hardness. SMCVs were calculated according to AlbertaEnvironmental Protection (1996). SMCVs varied for fish species from 0.00755 mg/L (rainbowtrout) to 0.060 mg/L (northern pike) and for invertebrate species from 0.006 mg/L (Daphniapulex) to 0.053 mg/L (Chironomus; Appendix 8).

The genus mean chronic values (GMCV) were calculated as outlined in Alberta EnvironmentalProtection (1996). The calculations are presented in Appendix 8 and result in a final chronicvalue (FCV) of 0.0095 mg/L. Two species had an SMCV lower than the final chronic value:Daphnia pulex with an SMCV of 0.006 mg/L and rainbow trout with an SMCV of 0.00755 mg/Lcopper. Because rainbow trout is a commercially, recreationally and ecologically important inAlberta, the final chronic value was adjusted to 0.007 mg/L copper to protect rainbow trout. Themost sensitive teratogenic effects were observed on rainbow trout embryos at copper valuesgreater than 0.01 mg/L (Birge and Black 1979). The FCV would therefore protect trout embryosfrom teratogenic effects and no adjustments to the FCV were therefore made. Figure 4 presents all primary chronic GMCVs in relation to the Alberta final chronic value.

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Final Plant Value

Many of the plant studies are unsuitable for derivation of the final plant value. Many differentgrowth media were used in the plant tests. Some of these media contained chelators, buffers andmetals that could potentially compete with copper for uptake by algae. These factors couldreduce toxicity to copper (Steemann-Nielsen et al. 1969; Ivorra et al. 1995). Only the primarystudies were rated: the studies without chelators, flow-through tests with measuredconcentrations, static tests without a significant decline in copper concentrations during the test,or static tests of short duration.

The only primary copper toxicity studies are with Staurastrum species (desmids; Ivorra et al.1995), Lemna minor (duckweed; Bishop and Perry 1981), Chlamydomonas reinhardtii (greenalga; Garvey et al. 1991), and Chlorella pyrenoidosa (green alga; Vavilin et al. 1995). In themedium without EDTA, the copper concentrations reducing 50% of growth (EC50) were 0.019

37

mg/L and less than 0.015 mg/L for Staurastrum chaetoceras and S. manfeldtii, respectively(Ivorra et al. 1995). Root length of Lemna minor was most affected by copper addition: EC50was 0.6 mg/L. Frond growth and growth rates were reduced 50% at 0.8 mg/L and 1.2 mg/L,respectively (Bishop and Perry 1981). Population growth of Chlamydomonas was not affected atcopper concentrations up to 0.064 mg/L. However, encystment and deflagellation were affectedat 0.048 mg/L and 0.009 mg/L, respectively (Garvey et al. 1991). A decrease of photosystem II(PSII) activity and tolerance to photoinhibition was observed in Chlorella pyrenoidosa at acopper concentration of 0.01 mg/L (Vavilin et al. 1995).

The final plant value is the lowest concentration with an important aquatic plant species resultingin a biologically significant effect. The lowest result was an LOEC value of 0.009 mg/L fordeflagellation of Chlamydomonas. Deflagellation however was not considered a biologicallysignificant endpoint. The next most sensitive value was the 0.01 mg/L LOEC value fordecreased PSII activity and decreased tolerance to photoinhibition. Although this test is not a"standard" toxicity endpoint, it is a measurement of a biologically significant process. The valueof 0.01 mg/L was therefore adopted as the final plant value.

Alberta Chronic Guideline

The Alberta chronic guideline is the lower of the final chronic value (0.007 mg/L) and the finalplant value (0.010 mg/L). The lowest value is the final chronic value; the Alberta chronicguideline is therefore 0.007 mg/L. This chronic guideline however should only be applied tosurface waters where hardness is equal to or greater than 50 mg/L CaCO3.

The chronic toxicity data span a water hardness range from 26 mg/L CaCO3 to over 200 mg /LCaCO3. However, most primary chronic toxicity values are performed in water with a hardnessbetween 50 and 200 mg/L CaCO3 (Appendix 8). The data in softer water (lower hardness) varyand are not conclusive. The restriction for using the copper guideline is added because:

• few data are available at water hardness below 50 mg/L CaCO3,• chronic toxicity should not be less stringent than acute toxicity (acute guideline would be

more stringent than the chronic guideline at water hardness less than 43 mg/L CaCO3).

This restriction will not greatly affect the use of the guideline in Alberta because surface water inAlberta is generally hard. Only a few lakes in specific regions of Jasper National Park, theCanadian Shield area (extreme northeast region of Alberta) and certain northern upland regions(Caribou and Birch Mountains) have low alkalinity and water hardness.

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4.3 Application of the Alberta Copper Guideline

Most of the toxicity studies cited in this document are carried out in laboratory water with lowcomplexing capacity. In contrast, surface water samples generally have much highercomplexation capacities than laboratory water. Total copper measurements in surface watertherefore consist largely of non-toxic forms (Sections 2.2 to 2.4). Little dissolved copper(containing the toxic copper species) is present and therefore only a small fraction of total copperin surface water would be actually toxic. Therefore total copper concentrations in surface waterbelow the copper guideline indicate a lack of copper toxicity. The toxic copper fraction would bea small fraction of total copper and would therefore be well below the copper guideline. Whentotal copper concentrations in surface water exceed the copper guideline the reverse argumenthowever cannot be made. A measurement of total copper provides no information on theconcentration of the toxic copper forms. In these cases, the concentration of the toxic copperforms could be either less or greater than the copper guideline. The following guidance istherefore provided to use and apply the copper guideline in an accurate and meaningful manner.

Preferred Approach

To properly asses the potential presence of copper toxicity, it is important to monitor the toxicform of copper in surface water. The best method to determine toxic forms is to analysedissolved copper in water (i.e., copper in a sample filtered through a 0.45 :m membrane filter). Other methods (copper ion electrode potentiometry and electrochemical methods) may determinethe more toxic copper forms compared to dissolved copper analyses. However, these methods donot include all the toxic copper species and are not routinely used in either toxicity tests (used toderive the copper guideline) or in ambient monitoring programs.

Although most copper in toxicity tests is added as a toxic, ionic form and laboratory water haslow complexing capacity, some of the copper changes during the test to a non-toxic form. Mosttoxicity endpoints used in the copper guideline are based on total copper or total acid-extractablecopper. A comparison of paired toxicity results expressed as total copper and dissolved copperindicated that in acute and chronic tests 52 to 95% of total copper was dissolved (Appendix J,USEPA 1994). A conversion factor of 0.96 is therefore recommended to adjust the total copperguideline to a dissolved copper guideline (USEPA. 1995. Interim Final Rule. Water QualityStandards. Establishment of Numeric Criteria for Priority Toxic Pollutants. State's Compliance -Review of Metals Criteria. Federal Register May 4, 1995. 40CFR Part 131). This conversionfactor is used to adjust the copper guidelines (expressed as total copper or acid-extractable)derived in Sections 4.1 and 4.2 to dissolved copper. Dissolved copper measurements in surfacewater determined by clean techniques should be compared to the following dissolved copperguidelines:

39

Dissolved Acute Guideline(mg/L)

Dissolved Chronic Guideline(mg/L)

at 100 mg/L CaCO3 0.015988*0.96=0.0153(0.015)

0.007544*0.96=0.0072(0.007)

at other hardness e0.979123ln(hardness)-8.68579 0.007

The use of clean techniques in determining dissolved copper concentrations is essential(Appendix L, USEPA 1994). These clean techniques prevent contamination of the dissolvedsample through proper sample collection, quality assurance and quality contol procedures, andlaboratory techniques. Clean techniques also prevent contamination of the filtered sample fromcopper contained in the membrane filters (filters should be soaked in acid until they are metal-free).

Alternative Approach

If clean techniques cannot be used to determine reliable and accurate dissolved copperconcentrations, acid-extractable copper measurement should be determined. This fraction ismore similar to copper measurements in the laboratory tests used to determine the copperguideline than total copper. These acid-extractable measurements should be compared to theguidelines calculated in Sections 4.1 and 4.2:

Acute Guideline (mg/L) e0.979123*ln(hardness)-8.64497

Chronic Guideline (mg/L) 0.007

As outlined above, environmental concentrations measured as acid-extractable copper exceedingthe Alberta guidelines may not indicate the presence of copper toxicity. For those situationswhere environmental concentrations exceed the Alberta guidelines, the possibility for negativeenvironmental effects could be determined further in two ways:

40

1 Use of clean techniques to determine the "true" dissolved copper concentrations. If thedissolved copper concentrations exceed the dissolved copper guideline, potential negativeeffects on instream biota may be present.

2 Determine a site-specific guideline as outline in the protocol for deriving Albertaguidelines for the protection of freshwater aquatic life (Alberta Environmental Protection1996). If the environmental concentrations exceed the site-specific guideline, negativeeffects on instream biota may be possible.

Soft Water

The chronic copper guideline does not apply to water with a hardness less than 50 mg/L CaCO3. Additional research is required to determine conclusively chronic toxicity of copper at lowhardness.

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5 DATA GAPS

Extensive information is available regarding the acute toxic effects of copper on aquatic biota. However, the information on chronic toxicity in soft water (water hardness less than 50 mg/LCaCO3) is limited and inconclusive. Additional research is required to improve the database andto allow development of a chronic guideline for soft water.

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6 COPPER GUIDELINES FROM OTHER JURISDICTIONS

Copper guidelines for the protection of freshwater aquatic life from other jurisdictions aresummarized in Table 7.

Table 7. Copper guidelines for the protection of freshwater aquatic life in otherjurisdictions.

Jurisdiction Type Guideline Hardness (H in mg/L CaCO3)

Alberta (1996) chronic 7 :g/L >50 mg/L

acute e0.979123*lnH-8.64497 mg/L

CCREM (1987) 2 :g/L0.2*e0.8545*ln(H)-1.465 :g/L

<60 mg/L>60 mg/L

Ontario (1994) chronic 5 :g/L

Saskatchewan (1988) 10 :g/L

British Columbia(Singleton 1987)

chronic 2 :g/L0.04*H :g/L

<50 mg/L >50 mg/L

acute 0.094*H+2 :g/L

USEPA (1984)/Quebec (1990)/Manitoba(Williamson 1988)

chronic e0.9422*ln(H)-1.464 :g/L

acute e0.8545*ln(H)-1.465 :g/L

FAO (Alabaster and Lloyd1982)

tentative 1 and 5 :g/L1

6 and 22 :g/L1

10 and 40 :g/L1

28 and 112 :g/L1

10 mg/L50 mg/L

100 mg/L300 mg/L

1 50th percentile and 95th percentile, respectively; adjustments can be made for thepresence of organic matter, low temperature, harmful substances and other species.

Because most guidelines vary with water hardness, a comparison of the various guidelineswith the Alberta guidelines at specific hardness values is presented in Table 8. The Albertaacute and chronic guidelines are similar to copper guidelines in other jurisdictions. However,the Alberta guidelines derived with the Alberta protocol are environmentally protective andpractical (Alberta Environmental Protection 1996). In addition, the Alberta guideline isbased on an up-to-date literature review and therefore is more current than other guidelines.

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Table 8. Comparison of freshwater aquatic life guidelines for copper (in :g/L) fromvarious jurisdictions.

GuidelineType

Jurisdiction Water Hardness (mg/L CaCO3)

50 100 200 300

Acute Alberta 8.1 16.0 31.5 46.8

USEPA/Quebec/Manitoba

9.2 17.7 34.0 49.9

British Columbia 7 11 20.8 30.2

Chronic Alberta 7 7 7 7

CCME 2 2.3 4.2 6.0

Ontario 5 5 5 5

USEPA/Quebec/Manitoba/

6.5 11.8 21.3 30.2

Saskatchewan 10 10 10 10

British Columbia 2 4 8 12

FAO 1/5 6/22 10/40 28/112

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76

Appendix 1. Acute and chronic copper toxicity data on amphibians.

Species Lifestage Test Type Toxicity Endpoint (mgCu/L)

Environmental Conditions Hardness Classification

Reference

Primary Studies

Two-linedsalamander

40 mm S 48-hr LC50 1.12 pH: measured; T: 20; Alk: 50-60;DO: 80% Sat

100-120 M, P Dobbs et al. 1994

Secondary Studies

Bufomelanosticus

tadpole, 1.8-2.2cm, 100 mg

S, CuSO4 24-hr LC50 0.84348-hr LC50 0.44696-hr LC50 0.32

pH: 7.4 (7.1-7.6); T: 29-34; Alk: 135(120-160); DO: 6.5 (5.8-7.8)

185 (165-215)

Nl, S Khangarot and Ray 1987a

Fowlers toad hatch4-d post-hatch

SR (2/d),CuSO4

3-d LC50 35.997-d LC50 26.96

pH: 7.2-7.8; T: 20-24; DO: near sat 100 (93-105) M-D, S Birge and Black 1979

Gray tree frog hatch4-d post-hatch

SR (2/d),CuSO4

3-d LC50 0.067-d LC50 0.04

pH: 7.2-7.8; T: 20-24; DO: near sat 100 (93-105) M-D, S Birge and Black 1979

Jeffersonsalamander

embryo S 96-hr LC50 0.315 pH: 4.5; T: 10; Alk: artificial softwater

M(NR), S Horne and Dunson 1994

Leopard frog hatch4-d post-hatch

SR (2/d),CuSO4

4-d LC50 0.068-d LC50 0.05

pH: 7.2-7.8; T: 20-24; DO: near sat 100 (93-105) M-D, S Birge and Black 1979

Marbledsalamander

hatch4-d post-hatch

SR (2/d),CuSO4

4-d LC50 3.598-d LC50 0.77

pH: 7.2-7.8; T: 20-24; DO: near sat 100 (93-105) M-D, S Birge and Black 1979

Microhylaornata

4-wk SR (1/d 24-hr LC50 6.0448-hr LC50 5.7496-hr LC50 5.38

pH: 6.86-6.94; T: 25.5-26; Alk: 97-98; DO: 8.2-8.4

142-145.5 N, S Rao and Madhyastha 1987

Microhylaornata

1-wk SR (1/d) 24-hr LC50 5.6148-hr LC50 5.3196-hr LC50 5.04

pH: 6.86-6.94; T: 25.5-26; Alk: 97-98; DO: 8.2-8.4

142-145.5 N, S Rao and Madhyastha 1987

Narrow-mouthed toad

hatch4-d post-hatch

SR (2/d),CuSO4

3-d LC50 0.057-d LC50 0.04 28-d LC50 0.09

pH 7.2-7.8; T: 20-24; DO: moderateaeration

197 M-D, S Birge and Black 1979

Rana tigrina 0.09 g S, CuSO4 96-hr LC50 0.389 pH 7.5 (7.4-7.8); T: 26.5 (24-27.5);Alk: 165 (140-190); DO: 6.6 (5.8-7.5)

240 (210-280)

N, S Khangarot et al. 1981

Xenopus laevis 3-4wk old larvae S 48-hr LC50 1.7 pH: 8+0.2; T: 20+1;Alk: 180; DO:9=0.5

N, S de Zwart and Slooff 1987

Species Lifestage Test Type Toxicity Endpoint (mgCu/L)

Environmental Conditions Hardness Classification

Reference

77

Unsuitable studies

Bufo boreas SR (1/2d),CuSO4

61-d NOEL 0.02 pH: 5-6; T: 19-21 NR, U Porter and Hakanson 1976

Rana pipiens adults SR (1/d) ,CuSO4

30-d LC50 0.0016% pH: 5.6; T: 20; Alk: tapwater not 100%aq, U

Kaplan and Yoh 1961

Ranahexadactyla

20 mm SR (1/d),CuSO4

96-hr LC50 0.039 pH: 6.2-6.7; T: 13-16; Alk: 24-40;DO: 6.2-7.0

13-80 N, U Khangarot et al. 1985

Rana pipiens embryo-larval S 72-hr LC50 0.15 pH: 7.73+0.5; T: 19.4+1.2;Alk:290+32; DO: 6.9+1.2

M-D, U Lande and Guttman 1973

Xenopus laevis female SR (1/d) 30-d NOEC 0.001%30-d LOEC 0.0015%

pH: 5.6; T: 22 dechlorinatedtapwater

N, U Fingal and Kaplan 1963

Legend: Test Type: S=Static, SR=Static Replacement, FT=Flow-Through;Environmental Conditions: T= Water Temperature oC, Alk=Alkalinity in mg/L CaCO3, DO=Dissolved Oxygen in mg/L unless otherwise

stated;Hardness: Water Hardness in mg/L CaCO3;Classification: M=Measured Concentrations, M-D= Measured Concentration, but significant decrease over test duration,

N=Nominal, NR=Not Reported; C=CalculatedP=Primary, S=Secondary, U=Unsuitable.

78

Appendix 2. Copper toxicity data on freshwater algae and plants.

Species Lifestage/Light

Test Type Toxicity Endpoint (mg Cu/L) Environmental Conditions Medium Classification Reference

Primary studies

Staurastrumchaetoceras

150 uE/m2/s 4-d EC50 0.0344-d EC50 0.7744-d EC50 0.019

pH: 7.8-8.4 T: 20pH: 7.0-7.1 T: 20pH: 7.0-7.1 T: 20

lakewatercomplete med.compl-EDTA

M,P Ivorra et al. 1995

Staurastrummanfeldtii

150 uE/m2/s 4-d EC50 0.0324-d EC50 0.4294-d EC50 <0.015

T: 20 lakewatercomplete med.compl-EDTA

M, P Ivorra et al. 1995

Chlamydomonasreinhardtii

28 Lux S, CuSO4 pop growth 72-hr LOEL>0.064encystment 72-hrNOEL 0.03472-hrLOEC 0.048deflagellation 72-hrNOEC 0.00572-hrLOEL 0.009

pH: 8;T: 20+1; Alk:59 ; Hardn: 76 no EDTA M, P Garvey et al. 1991

Chlorellapyrenoidosa

120 umol/m2/s CuSO4 PSII act. 12-hr LOEC 0.010 pH: 6.8; T: 35 Tanuya med.-EDTA

N, P Vavilin et al. 1995

Lemna minor FT,CuSO4

7-d EC50 root length 0.6 pH: 7.2-7.6; Hard: 120-130 8.5 M, P Bishop and Perry 1981

Non-primary studies

Ankistrodesmusfalcatus

140 ft candle S 14-d LOEL growth 0.25 pH: 8-8.9; T: 22.2; Alk:45-50;Hard: 45-50 Gerloff's Chu 2RP, N Maloney and Palmer 1956

Ankistrodesmusfalcatus, acicularis

140 ft candle S 14-d LOEL growth 0.51 pH: 8-8.9; T: 22.2; Alk:45-50; Hard: 45-50 Gerloff's Chu 2RP, N Maloney and Palmer 1956

Ankistrodesmusbraumii

8x104 cells/mL CuNO3 20-d growth NOEL 0.00620-d growth LOEL 0.063

T: 20 Ohad (1967) N Laube et al. 1980

Chlamydomonascommunis

140 ft candle S 14-d LOEL growth 0.51 pH: 8-8.9; T: 22.2; Alk:45-50; Hard: 45-50 Gerloff's Chu 2RP, N Maloney and Palmer 1956

Chlamydomonasparadoxa

140 ft candle S 14-dLOEL growth 0.25 pH: 8-8.9; T: 22.2; Alk:45-50; Hard: 45-50 Gerloff's Chu 2RP, N Maloney and Palmer 1956

Chlorella variegata 140 ft candle S 14-d LOEL growth 0.25 pH: 8-8.9; T: 22.2; Alk:45-50; Hard: 45-50 Gerloff's Chu 2RP, N Maloney and Palmer 1956

Chlorella vulgaris 1x105 cells/L S ? photosynthesis EC50 2.3 pH: 6.1-6.3; T: 30 N Hassall 1963

Species Lifestage/Light

Test Type Toxicity Endpoint (mg Cu/L) Environmental Conditions Medium Classification Reference

79

Chlorella vulgaris 8000 Lux S, CuSO4 growth to stat. phase LOEC 0.05 pH: 6.2; T: 24 Art. -EDTA 3RP, NR Foster 1977

Chlorella ellipsoida

division NOEL 0.02 pH: 5.6 start; T: 23 Arnon A5 N Kanazawa and Kanazawa 1969

Chlorella fusca S survival 0.095 mg/L 10% Bristol NR Wong 1989

Chlorella fusca 104 cells/mL CuSO4 20-d growth LOEL 0.2 mg/L T: 18-20 Bristol N Wong and Beaver 1980

Chlorellapyrenoidosa

1.8x107 cells/L S 20-hr photosynthesis EC49 0.05 pH: 8; T: 20 Osterlind N Steeman-Nielsen et al. 1969

Chlorellapyrenoidosa

4x104 cells/mL S 72-hr cell div. IC50 1672-hr cell div. IC50 2472-hr cell div. IC50 >200

T: 21; enr. soft waterT: 21; EPA mediumT: 21; MBL medium

N Stauber and Florence 1989

Chlorella sp. 20kLux,8.3x106

cells/mL

S, CuNO3 4-hr LOEL 0.006 pH: 7 Nutrient N Gachter et al. 1973

Chlorella sp. 6/7 Lux 100-150 hr 0.005 T: 20 Osterlind B-EDTA Steeman-Nielsen and Kamp-Nielsen 1970

Chlorella sp. 165.4 uE/m2/s CuSO4 96-hr NOEL 2.596-hr LOEL 25

pH: 22 BG11 N Brady et al. 1994

Chlorococcumbotryoides

140 ft candle S 14-d LOEL growth 1.02 pH: 8-8.9; T: 22.2; Alk:45-50; Hard: 45-50 Gerloff's Chu 2RP, N Maloney and Palmer 1956

Chlorococcumhumicola

140 ft candle S 14-d LOEL growth 0.127 pH: 8-8.9; T: 22.2; Alk:45-50; Hard: 45-50 Gerloff's Chu 2RP, N Maloney and Palmer 1956

Coccomyxa simplex

140 ft candle S 14-d LOEL growth 0.06 pH: 8-8.9; T: 22.2; Alk:45-50; Hard: 45-50 Gerloff's 2RP, N Maloney and Palmer 1956

Coelastrumprobscideum

140 ft candle S 14-d LOEL growth 0.51 pH: 8-8.9; T: 22.2; Alk:45-50; Hard: 45-50 Gerloff's Chu 2RP, N Maloney and Palmer 1956

Gloeocystisgrevillei

140 ft candle S 14-d LOEL growth 0.127 pH: 8-8.9; T: 22.2; Alk:45-50; Hard: 45-50 Gerloff's Chu 2RP, N Maloney and Palmer 1956

Mesotaeniumcaldariorum

140 ft candle S 14-d LOEL growth 0.51 pH: 8-8.9; T: 22.2; Alk:45-50; Hard: 45-50 Gerloff's Chu 2RP, N Maloney and Palmer 1956

Oocystis lacustris 140 ft candle S 14-d LOEL growth 0.51 pH: 8-8.9; T: 22.2; Alk:45-50; Hard: 45-50 Gerloff's Chumedium

2RP, N Maloney and Palmer 1956

Species Lifestage/Light

Test Type Toxicity Endpoint (mg Cu/L) Environmental Conditions Medium Classification Reference

80

Oocystis marsonii 140 ft candle S 14-d LOEL growth 0.51 pH: 8-8.9; T: 22.2; Alk:45-50; Hard: 45-50 Gerloff's Chu 2RP, N Maloney and Palmer 1956

Scenedesmusquadricauda

2.6x106 cells/L S, CuSO4 14-d no irreversible damage 0.8-1.0 T: 20 Chu No.10 N Gibson 1972

Scenedesmusquadricauda

S, CuCl2 10-d EC33 1.0 T: 20-22 N Khobot'yev et al. 1975

Scenedesmusquadricauda

S 300-hr 2 ng/L pH: 7.55; T: 23 Art+EDTA M,C Verwey et al. 1992

Scenedesmusquadricauda

S survival 0.09 mg/L 10% Bristol ? Wong 1989

Scenedesmusquadricauda

5x104 cells/mL CuCl2 21-d growth LOEL 0.05 mg/L T: 18-20 Bristol N Wong and Beaver 1981

Scenedesmusbasilensis

140 ft candle S 14-d LOEL growth 0.51 pH: 8-8.9; T: 22.2; Alk:45-50; Hard: 45-50 Gerloff's Chu 2RP, N Maloney and Palmer 1956

Scenedesmisobliquus

140 ft candle S 14-d LOEL growth 4.1 pH: 8-8.9; T: 22.2; Alk:45-50; Hard: 45-50 Gerloff's Chu 2RP, N Maloney and Palmer 1956

Scenedesmuscapricornutum

1.2x106

cells/mLS,CuSO4 1-3 hr NOEL 0.05 mg/L

1-3 hr LOEL 0.15 mg/LpH: 6.8 min, 15 mg/L

NaHCO33RP,N Arsenault et al. 1993

Scenedesmus sp. 165.4 uE/m2/s CuSO4 96-hr NOEL 2596-hr LOEL 100

T: 22 BG11 N Brady et al. 1994

Selenastrumcapricornutum

21-d EC50 cell vol. 0.085 SAAM N Christensen and Scherfig 1979

Selenastrumcapricornutum

104 cells/mL EC50 0.0476 pH: increased to 8.5 shaken 100rpm NR Christensen and Nyholm 1984

Selenastrumcapricornutum

CuCl2 4-7 d incipient inh. 0.05 pH: 7.1-7.2; T: 24; Alk:8.2; Hard: 14.9 agitated N Bartlett et al. 1974

Selenastrumcapricornutum

S/SR,CuSO4

72-hr grwoth EC50 0.054/0.048 T: 24+2 LC/ISO 3RP, N Radetski et al. 1995

Selenastrum sp. S survival 0.09 mg/L 10% Bristol NR Wong 1989

Selenastrum sp. 165.4 uE/m2/s CuSO4 96-hr NOEL 25 mg/L96-hr LOEL 100

T: 22 BG11 N Brady et al. 1994

Species Lifestage/Light

Test Type Toxicity Endpoint (mg Cu/L) Environmental Conditions Medium Classification Reference

81

Sphaerella lacustris

140 ft candle S 14-d LOEL growth 0.25 pH: 8-8.9; T: 22.2; Alk:45-50; Hard: 45-50 Gerloff's Chu 2RP, N Maloney and Palmer 1956

Stigeocloniumnanum

140 ft candle S 14-d LOEL growth 0.127 pH: 8-8.9; T: 22.2; Alk:45-50; Hard: 45-50 Gerloff's Chumedium

2RP, N Maloney and Palmer 1956

Anabaena flos-aquae

S 10-d no irreversible damage 0.1-0.25

T: 20 T: 20 N Gibson 1972

Anabaena flos-aquae

OD 0.01OD0.2

CuSO4 nitrogen fixation16-hr IC90 0.0051-hr IC50 0.32

pH: 8 inorg. N Peterson et al. 1995

Anabaena variabilis 10-d EC90 1.0 T: 20-22 N Kobot'yev et al. 1975

Anabaenacylindrica

rate of movement EC50 0.5E100 4 mg/L

NR Fogg and Westlake 1953

Anabaena 7120 8x104 cells/mL CuNO3 growth20-d NOEL 0.00620-d LOEL 0.063

T: 20 BGII medium N Laube et al. 1980

Calothrix braunii 140 ft candle S 14-d LOEL growth 4.07 pH: 8-8.9; T: 22.2; Alk:45-50; Hard: 45-50 Gerloff's Chumedium

2RP, N Maloney and Palmer 1956

Cylindrospermumlicheniforme

140 ft candle S 14-d LOEL growth 0.127 pH: 8-8.9; T: 22.2; Alk:45-50; Hard: 45-50 Gerloff's Chumedium

2RP, N Maloney and Palmer 1956

Microcystisaeruginosa

140 ft candle S 14-d LOEL growth 0.127 pH: 8-8.9; T: 22.2; Alk:45-50; Hard: 45-50 Gerloff's Chumedium

2RP, N Maloney and Palmer 1956

Nostoc mescorum 140 ft candle S 14-dLOEL growth 0.51 pH: 8-8.9; T: 22.2; Alk:45-50; Hard: 45-50 Gerloff's Chumedium

2RP, N Maloney and Palmer 1956

Phormidium tenue 140 ft candle S 14-d LOEL growth 0.127 pH: 8-8.9; T: 22.2; Alk:45-50; Hard: 45-50 Gerloff's Chumedium

2RP, N Maloney and Palmer 1956

Plectonemanostocorum

140 ft candle S 14-d LOEL growth 0.127 pH: 8-8.9; T: 22.2; Alk:45-50; Hard: 45-50 Gerloff's Chumedium

2RP, N Maloney and Palmer 1956

Symploca erecta 140 ft candle S 14-d LOEL growth 1.02 pH: 8-8.9; T: 22.2; Alk:45-50; Hard: 45-50 Gerloff's Chumedium

2RP, N Maloney and Palmer 1956

Achanthes linearis#1

140 ft candle S 14-d LOEL growth 0.51 pH: 8-8.9; T: 22.2; Alk:45-50; Hard: 45-50 Gerloff's Chumedium

2RP, N Maloney and Palmer 1956

Species Lifestage/Light

Test Type Toxicity Endpoint (mg Cu/L) Environmental Conditions Medium Classification Reference

82

Achanthes linearis#2

140 ft candle S 14-d LOEL growth 0.25 pH: 8-8.9; T: 22.2; Alk:45-50; Hard: 45-50 Gerloff's Chumedium

2RP, N Maloney and Palmer 1956

Gomphonemaparvulum

140 ft candle S 14-d LOEL growth 0.06 pH: 8-8.9; T: 22.2; Alk:45-50; Hard: 45-50 Gerloff's Chumedium

2RP, N Maloney and Palmer 1956

Nitzschia palea #1 140 ft candle S 14-d LOEL growth 0.127 pH: 8-8.9; T: 22.2; Alk:45-50; Hard: 45-50 Gerloff's Chumedium

2RP, N Maloney and Palmer 1956

Nitzschia palea #2 140 ft candle S 14-d LOEL growth 0.51 pH: 8-8.9; T: 22.2; Alk:45-50; Hard: 45-50 Gerloff's Chumedium

2RP, N Maloney and Palmer 1956

Nitzschia palea #3 140 ft candle S 14-d LOEL growth 0.127 pH: 8-8.9; T: 22.2; Alk:45-50; Hard: 45-50 Gerloff's Chu medium 2RP, N Maloney and Palmer 1956

Nitzschia linearis S, CuSO4 growth 120-hr IC50 0.795-0.815 synthetic medium N Patrick et al. 1968

Elodea canadensis 16,00 lux S O2 evolution 48-hr IC50 0.15 T: 24 HEPES buffer N Brown and Rattigan 1979

Lemna minor S 14-d plant damage 0.13 HEPES buffer N Brown and Rattigan 1979

Hydrilla verticillata 7-10 cm shootlength

CuSO4 growth LOEL 1growth NOEL 0.1POD activity 5-d LOEL 0.015-d NOEL 0.001

pH: 6; T: 25 Hoagland +EDTA N Byl et al. 1994

Anabaena flos-aque S CuSO4

growth90-d LOEL 0.290-d NOEL 0.1

T: 20 Medium D N Young and Lisk 1972

Scenedesmusquadricauda

7-d TT 1.1 T: 27 Bringmann and Kuhn 1980

Chlorella vulgaris CuCl2 33d growth EC50 0.18 pH: 7+0.1; T: 21+1 Bristol N Rosko and Rachlin 1977

Nostoc muscorum 100uE/m2/s 5min PSII inhibit. EC50 0.635growth 0.19

T: 24+1 Pbuffer,sucrose,NaCl N Mishra et al. 1993

Lemna minor 80 umol/m2/s laminar flowsystem

surf.cov. 14-d EC50 0.14multiplic. 14-d EC50 0.32

pH: 5+0.1; T: 23+1 Gorham+2.5uM EDTA

M Jenner and Janssen-Mommen 1993

Cylindrospermum 10 W/m2 multipl. NOEL 0.06 T: 24+1 modified Chu N Khare and Bisen 1991

Myriophyllum spicatum 300 fc S root weight 32-d IC50 0.25 T: 20 synthetic N Stanley 1974

Amphora coffaeformis

130 umol/m2/s SCu2O/CuCl2

96-hr EC50 6.4-7.6MIC4.2-2.9

pH: 8.1; T: 20 Guillard f/2 (chelators) M-D French and Evans 1988

Species Lifestage/Light

Test Type Toxicity Endpoint (mg Cu/L) Environmental Conditions Medium Classification Reference

83

Amphora hyalina 130 umol/m2/s SCu2O/CuCl2

96-hr EC50 5.8-3.5MIC 5.0-5.3

pH: 8.1; T: 20 Guillard f/2 (chelators) M-D French and Evans 1988

Selenastrumcapricornutum

EC50 0.042 Porcella EPA method N Miller et al. 1985

Chlorella vulgaris growth 3-4 mo NOEL 0.023-4 mo LOEL 0.04

pH: 7; T: room temp inorg. minimum med N Den Dooren 1965

Chlorella vulgaris S 96-hr growth EC50 0.2 T: 20+1 OECD M-D Blaylock et al. 1985

Selenastrumcapricornutum

S 96-hr growth EC50 0.4 T: 20+1 OECD M-D Blaylock et al. 1985

Anacystis nidulans NOEC 0.2/2LOEC 2/20

+/- EDTA N Lee et al. 1993

Anacystis nidulans S, CuCl2 6-d PSIIact. LOEC 0.318 pH: 7.25; T: 25 Hepes+pBQ N Gupta and Singhal 1995

Chroococcis paris 10-d growth LOEC 0.1 T: 26 BG11 N Les and Walker 1984

Lemna paucicostata 6000 Lux CuSO4 12-d growth EC30 0.3 pH: 5.1; T: 25+1 Hoagland N Nasu and Kugimoto 1981

Legend: Test Type: S=Static, SR=Static Replacement, FT=Flow-Through;Environmental Conditions: T= Water Temperature oC, Alk=Alkalinity in mg/L CaCO3, DO=Dissolved Oxygen in mg/L unless otherwise stated;

Hard=Water Hardness in mg/L CaCO3;Classification: M=Measured Concentrations, M-D= Measured Concentration, but significant decrease over test duration,

N=Nominal, NR=Not Reported; C=Calculated; RP=ReplicatedP=Primary, S=Secondary, U=Unsuitable.

84

Appendix 3. Acute copper toxicity data on freshwater fish.

Species Lifestage Test Type Toxicity Endpoint (mg Cu/L) Environmental Conditions Hardness Classification Reference

Primary Studies

Atlantic salmon 8.8-9.8 cm FT,CuSO4

96-hr ILL 0.025 T: 3.8-4.8; DO: light aeration 14 M, P Carson and Carson 1972

Atlantic salmon 9.2 (7.2-10.9)cm

FT 48-hr ILL 0.032 pH: 7.3 (7-7.4); T: 17 14 M, P Sprague and Ramsay1965

Atlantic salmon 8.5 (6.4-11.7)cm

FT 200 hr ILL 0.048 pH: 7.1-7.5; T: 15; Alk: 12; DO: diffused air 20 M, P Sprague 1964b

Blacknose dace 47 mm FT, CuSO4 96-hr LC50 0.32 pH: 7.9-8.1; T: 23-25; Alk: 148-161 196-205 M, P Geckler et al. 1976

Bluegill 35.8+1 mm FT, CuCl2 96-hr LC50 1.3 Alk: 82 85 M, P Blaylock et al. 1985

Bluegill 49mm FT, CuCl2 96-hr LC50 0.9-1.1 pH: 6.8-7.5; T: 22+1; Alk: 23.2-32.8; DO: 6.6-9.5

21.2-59.2 M, P Thompson et al. 1980

Bluegill 35 g, 12 cm FT, CuSO4 96-hr LC50 1.1 mg/L pH: 7-8; T: 20+1; Alk: 43+1.1; DO: 7+1.2 45+0.9 M, P Benoit 1975

Bluegill 35mm S 48-hr LC50 4.3 pH: 7.5-7.8; T: 20; Alk: 50-60; DO: 80% Sat 100-120 M, P Dobbs et al. 1994

Bluegill 1-9 g S,CuSO4/CuCl2

96-hr LC50 0.74 mg/L pH: 4.5-6.5 (pH control); T: 20; Alk: 3-6; DO:>4.5 mg/L

46 M, P Trama 1954

Bluntnose minnow FlT,CuSO4

96-hr LC50 0.23 pH: 7.88-8.31; T: 25; DO: >5.9 mg/L 200 M, P Horning and Neiheisel1979

Bluntnose minnow 84 mm FT, CuSO4 96-hr LC50 0.34 pH: 7.9-8.1; T: 23-25; Alk: 148-161 196-205 M, P Geckler et al. 1976

Brook trout 14 mo old FT 96-hr LC50 0.100 pH: 7.5; T: 12; Alk: 42 45 M, P McKim and Benoit 1971

Brown bullhead 7 mo2 yr

FT, CuSO4 96-hr LC50 0.17-0.1996-hr LC50 0.186

pH: 7.6; T: 23; Alk: 156; DO: 6.6+1.9pH: 7.6; T: 15-20; Alk: 156; DO: 6.6+1.9

202 M, P Brungs et al. 1973

Brown bullhead 39 mm FT, CuSO4 96-hr LC50 0.54 pH: 7.9-8.1; T: 23-25; Alk: 148-161 196-205 M, P Geckler et al. 1976

Checkeredrainbowfish

juveniles FT 96-hr LC50 0.168-0.19 pH: 5.85-6.15; T: 27+0.5; Alk: 5 25-30 M, P Williams et al. 1991

Chinook salmon 3.9-6.8 cm FT, CuSO4 96-hr LC50 0.032 pH: 7.3-7.0; T: 11-13; DO: 90% Sat 20-22 M, P Finlayson and Verrue1982

Species Lifestage Test Type Toxicity Endpoint (mg Cu/L) Environmental Conditions Hardness Classification Reference

85

Chinook salmon 1.35 g FT 96-hr LC50 0.0196-hr LC50 0.02596-hr LC50 0.0996-hr LC50 0.125

pH: 7.2;T: 12; Alk: 12; DO: measuredpH: 7.6; T: 12; Alk: 35 ; DO: measuredpH: 8.1; T: 12; Alk: 125; DO: measuredpH: 8.5; T: 12; Alk: 243; DO: measured

13 46182359

M, P Chapman and McCrady1977

Chinook salmon Alevin 0.05gSwim-p 0.23gParr 11.58gSmolt 32.46g

FT 96-hr LC50 0.02696-hr LC50 0.01996-hr LC50 0.03896-hr LC50 0.026

pH: 7.1; T: 12.2+0.4; Alk: 22+2; DO: 10.2+0.3 23+1 M, P Chapman 1978

Chiselmouth 1.25 g FT, CuCl2 96-hr LC50 0.143 pH: 7.1-7.5; T: 10.5; Alk: 20-30; DO: 8.6-11.3 20-30 M, P Andros and Garton 1980

Coho salmon yearling SR, CuCl2 96-hr LC50 0.06-0.074 pH: 6.8-7.9; T: 10-12; Alk: 68-78; DO: aerated(6-10.8 mg/L)

95 (89-99) M, P Lorz and McPherson1976

Creek chub 64 mm FT, CuSO4 96-hr LC50 0.31 pH: 7.9-8.1; T: 23-25; Alk: 148-161 196-205 M, P Geckler et al. 1976

Cutthroat trout 5.7 g FT, CuCl2 96-hr LC50 0.0157 pH: 7.64; T: 9.8; Alk: 20.1 (19-24); DO: 7.7(6.7-8.9)

26.4 (23-30) M, P Chakoumakos et al. 1979

Cutthroat trout 4.2 g FT, CuCl2 96-hr LC50 0.367 pH: 7.73; T: 9.8; Alk: 178 (173-188); DO: 7.7(6.7-8.9)

205 (202-208) M, P Chakoumakos et al. 1979

Eel-tailed catfish juveniles FT 72-hr LC50 0.085 pH: 5.85-6.15; T: 27+0.5; Alk: 5 25-30 M, P Williams et al. 1991

Fathead minnow 1-d FT,CuSO4

96-hr LC50 (T/D/CU2+)0.008/0.007/0.00095

pH: 6.58; T: 22; Alk: 48.9 49.2 M, P Nelson et al. 1986

Fathead minnow 1-d FT,CuSO4

96-hr LC50 (T/D/CU2+)0.075/0.070/0.0037

pH: 8.12; T: 22; Alk: 43.3 47.2 (NaCl2) M, P Nelson et al. 1986

Fathead minnow 6-mo old 6-wk old

FT 96-hr LC50 0.4696-hr LC50 0.49

pH: 7.5-8.2; T: avg 23-24; DO: 7.8+0.74 202 M, P Pickering et al. 1977

Fathead minnow 10-20mm FT 96-hr LC50 0.075 pH: 6.9-7.2; T: 25; Alk: 30-31; DO: 7.2-7.9 31 M, P Mount and Stephan 1969

Fathead minnow 56 mm47 mm

FT, CuSO4 96-hr LC50 0.4496-hr LC50 0.49

pH: 7.9-8.1; T: 23-25; Alk: 148-161 196-205 M, P Geckler et al. 1976

Fathead minnow 3 wk old S 48-hr LC50 0.284 pH: 7.5-7.8; T: 20; Alk: 50-60; DO: 80% Sat 100-120 M, P Dobbs et al. 1994

Fathead minnow 1-d FT,CuSO4

96-hr LC50 (T/D/CU2+)0.079/0.056/0.0013

pH: 7.94; T: 22; Alk: 148 45 M, P Nelson et al. 1986

Species Lifestage Test Type Toxicity Endpoint (mg Cu/L) Environmental Conditions Hardness Classification Reference

86

Fathead minnow 1-d FT,CuSO4

96-hr LC50 (T/D/CU2+)0.048/0.040/0.0016

pH: 8.06; T: 22; Alk: 43.3 255 (MgCl2) M, P Nelson et al. 1986

Fathead minnow 1-d FT,CuSO4

96-hr LC50 (T/D/CU2+)0.119/0.109/0.0054

pH: 8.05; T: 22; Alk: 43.1 243 (CaCl2) M, P Nelson et al. 1986

Fathead minnow 1-d FT,CuSO4

96-hr LC50 (T/D/CU2+)0.060/0.050/0.0020

pH: 8.06; T: 22; Alk: 42.8 45.1 M, P Nelson et al. 1986

Fathead minnow 1-d FT,CuSO4

96-hr LC50 (T/D/CU2+)0.157/0.136/0.00079

pH: 8.57; T: 22; Alk: 150 45 M, P Nelson et al. 1986

Fathead minnow 1-d FT,CuSO4

96-hr LC50 (T/D/CU2+)0.022/0.021/0.0043

pH: 7.16; T: 22; Alk: 26.4 45 M, P Nelson et al. 1986

Fathead minnow 1-d FT,CuSO4

96-hr LC50 (T/D/CU2+)0.068/0.046/0.0053

pH: 7.84; T: 22; Alk: 17 44 M, P Nelson et al. 1986

Fathead minnow 1-d FT,CuSO4

96-hr LC50 (T/D/CU2+)0.095/0.079/0.004

pH: 7.93; T: 22; Alk: 42 44 M, P Nelson et al. 1986

Fathead minnow 1-d FT,CuSO4

96-hr LC50 (T/D/CU2+)0.022/0.019/0.0021

pH: 7.15; T: 22; Alk: 155 46.2 M, P Nelson et al. 1986

Fathead minnow 1-d FT,CuSO4

96-hr LC50 (T/D/CU2+)0.032/0.028/0.00057

pH: 8.13; T: 22; Alk: 42.6 47 M, P Nelson et al. 1986

Fathead minnow S, CuCl2 96-hr LC50 0.055 pH: 7.7; T: <22-28; Alk: 55; DO: 7.4-8.3 52 M, P Carlson et al. 1986

Fathead minnow 30 d, 0.19 g FT,CuNO3

96-hr LC50 0.096 pH: 7.4; T: 25+3;Alk: 42.4+1.9; DO: >70% sat. 43.9+1 M,P Spehar and Fiandt 1986

Fathead minnow 1-d FT,CuSO4

96-hr LC50 (T/D/CU2+)0.051/0.039/0.00067

pH: 7.98; T: 22; Alk: 161 44 M, P Nelson et al. 1986

Fathead minnow 1-d FT,CuSO4

96-hr LC50 (T/D/CU2+)0.023/0.019/0.0019

pH: 7.35; T: 22; Alk: 44.5 46.2 M, P Nelson et al. 1986

Fathead minnow 1-d FT,CuSO4

96-hr LC50 (T/D/CU2+)0.082/0.071/0.00056

pH: 8.73; T: 22; Alk: 47.4 45.2 M, P Nelson et al. 1986

Fathead minnow 1-d FT,CuSO4

96-hr LC50 (T/D/CU2+)0.066/0.046/0.0004

pH: 8.00; T: 22; Alk: 318 45 M, P Nelson et al. 1986

Fathead minnow 25-35 d 48-hr LC50 0.18 pH: 7.5-8.5; T: 20+2; Alk: 30-45 45-60 M, P Jop et al. 1993

Fathead minnow <24 hr FT,CuSO4

96-hr LC50 0.074 (0.042-0.0107)

pH:8.1; T: 22; DO >90% Sat. 45 M, P Erickson et al. 1996

Species Lifestage Test Type Toxicity Endpoint (mg Cu/L) Environmental Conditions Hardness Classification Reference

87

Flagfish 0.1-0.3 g FT 96-hr LC50 1.27 pH: 8.0-8.3; T: 25; Alk: 200-255; DO: avg 75%sat

350-375 M, P Fogels and Sprague 1977

Goldfish 3.1-6 cm FT, CuSO4 96-hr LC50 0.3 pH: 7.33 (7.1-9.3); T: 21 (20-22.5); DO: 7.47(5.05-8.4)

52 (45-96) M, P Tsai and McKee 1980

Guppies 10-21 d FT 96-hr LC50 0.11296-hr LC50 0.138

pH: 7.6; T: 25; Alk: 31pH: 7.4; T: 25.9; Alk: 36

67

88

M, P Chynoweth et al. 1976

Guppies 0.12-0.29 g LC50 0.16-0.31 mg/L pH: 7.0; T: 25; Alk: 124 144 M, P Anderson and Weber1975

Guppy 0.1 g FT, CuCl2 96-hr LC50 0.139 pH: 7+0.1; T: 25+0.5; Alk: 144; DO: 99% sat 124 M, P Spear and Anderson1975

King salmon 0.3 g FT 10-d after hatch LC50 0.021-0.04

pH: 6.8-7.2; T: 13-14; Alk: 21; DO: nearsaturation

44 M, P Hazel and Meith 1970

Northern squawfish 1.33 g FT, CuCl2 96-hr LC50 0.02396-hr LC50 0.018

pH: 7.1-7.5; T: 7.8; Alk: 20-30; DO: 8.6-11.3pH: 7.1-7.5; T: 11.5; Alk: 20-30; DO: 8.6-11.3

20-30 M, P Andros and Garton 1980

Orangethroat darter 44 mm FT, CuSO4 96-hr LC50 0.85 pH: 7.9-8.1; T: 23-25; Alk: 148-161 196-205 M, P Geckler et al. 1976

Pennyfish juveniles FT 96-hr LC50 0.077 pH: 5.85-6.15; T: 27+0.5; Alk: 5 25-30 M, P Williams et al. 1991

Pink salmon newly hatchedalevinfry

96-hr LC50 0.14396-hr LC50 0.08796-hr LC50 0.199

pH: 7.6 (7.55-7.8); T: 5.5-8.9; Alk: 62.5 (60.8-68.2); DO: Saturated

83.1 (82.9-84.4)

M, P Servizi and Martens1978

Pumpkinseeds 2.2 g FT, CuCl2 96-hr LC50 1.30 pH: 7.2+0.2; T: 20+1.0; Alk: 85+2; DO: 79-89% sat

125+5 M, P Spear and Anderson1975

Pumpkinseeds 1.2 g FT, CuCl2 96-hr LC50 1.24 pH: 7.2+0.2; T: 20+1.0; Alk: 85+2; DO: 79-89% sat

125+5 M, P Spear and Anderson1975

Pumpkinseeds 7.6 g FT, CuCl2 96-hr LC50 1.94 pH: 7.2+0.2; T: 20+1.0; Alk: 85+2; DO: 79-89% sat

125+5 M, P Spear and Anderson1975

Pumpkinseeds 4.5 g FT, CuCl2 96-hr LC50 1.67 pH: 7.2+0.2; T: 20+1.0; Alk: 85+2; DO: 79-89% sat

125+5 M, P Spear and Anderson1975

Rainbow trout alevin SR (1/2d) 144-hr LC50 0.018144-hr LC50 0.019

pH: 5.8; T: 8.6; Alk: 3.6pH: 4.9; T: 8.6; Alk: 3.6

10.3 M, P Hickie et al. 1993

Rainbow darter 41 mm FT, CuSO4 96-hr LC50 0.32 pH: 7.9-8.1; T: 23-25; Alk: 148-161 196-205 M, P Geckler et al. 1976

Species Lifestage Test Type Toxicity Endpoint (mg Cu/L) Environmental Conditions Hardness Classification Reference

88

Rainbow trout embryo 12mm SR (1/d) 96-hr LC50 0.4 pH: 7.3-7.7; T: 10; Alk: softwater Stephan 197546-48; DO: 9.2-9.5

M, P Giles and Klaverkamp1982

Rainbow trout 176 g FT, CuCl2 96-hr LC50 0.21 pH: 7.8+0.1; T: 10+1.0; Alk: 85+2; DO: 74-95% sat

125+5 M, P Spear and Anderson1975

Rainbow trout 28.5 g SR (6-hr) 48-hr LC50 0.75 pH: 7.3-7.5; T: 15.3-18.4 (+0.5); DO: >90% sat 240 M, P Brown and Dalton 1970

Rainbow trout 1.2-7.9 g FT 96-hr LC50 0.102 pH: 8.0-8.3; T: 15; Alk: 200-255; DO: avg 75%sat.

350-375 M, P Fogels and Sprague 1977

Rainbow trout 13-15 cm FT 72-hr LC50 0.58 pH: 7.35-7.6; T: 12-15; DO: 80-90% sat. 250 M, P Brown et al 1974

Rainbow trout 12-16 cm FT 96-hr LC50 0.8914-d LC50 0.87

pH: 7.3-7.4; T: 15-15.6; Alk: 200-210; DO:>70% sat

290-310 M, P Calamari and Marchetti1973

Rainbow trout 8-15 g FT 15-d LC50 0.048 pH: 7.3;T: 13+1; Alk: 28 (24-31); DO: wellaerated

49 (46-54) M, P Miller and Mackay 1980

Rainbow trout 3.9 g FT, CuCl2 96-hr LC50 0.200 pH: 7.8+0.1; T: 10+1.0; Alk: 85+2; DO: 74-95% sat

125+5 M, P Spear and Anderson1975

Rainbow trout 29.1 g FT, CuCl2 96-hr LC50 0.19 pH: 7.8+0.1; T: 10+1.0; Alk: 85+2; DO: 74-95% sat

125+5 M, P Spear and Anderson1975

Rainbow trout juvenile FT,CuSO4

6-d LC50 0.274-0.381 pH: 7.7-7.9; T: 14.9-15.0; Alk: 212-236; DO:7.8-9.2

346-386 M, P Dixon 1980

Rainbow trout norm to 10 g fish FT, CuSO4 96-hr LC50 0.39396-hr LC50 0.15496-hr LC50 0.34496-hr LC50 0.82596-hr LC50 0.167

pH: 5; T: 15; Alk: 170; DO: >90% sat.pH: 6; T: 15; Alk: 170; DO: >90% sat.pH: 7; T: 15; Alk: 170; DO: >90% sat.pH: 8; T: 15; Alk: 170; DO: >90% sat.pH: 9; T: 15; Alk: 170; DO: >90% sat.

370369361366364

M, P Howarth and Sprague1978

Rainbow trout norm to 10 g fish FT, CuSO4 96-hr LC50 0.34996-hr LC50 0.05996-hr LC50 0.08596-hr LC50 0.191

pH: 4.9; T: 15; Alk: 84; DO: >90% sat.pH: 6; T: 15; Alk: 84; DO: >90% sat.pH: 7; T: 15; Alk: 84; DO: >90% sat.pH: 9; T: 15; ; DO: >90% sat.

10110110098

M, P Howarth and Sprague1978

Rainbow trout norm to 10 g fish FT, CuSO4 96-hr LC50 0.03096-hr LC50 0.04996-hr LC50 0.05996-hr LC50 0.047

pH: 5; T: 15; Alk: 36; DO: >90% sat.PH: 6; T: 15; Alk: 36; DO: >90% sat.pH: 8; T: 15; Alk: 36; DO: >90% sat.pH: 9; T: 15; Alk: 36; DO: >90% sat.

3031.53130

M, P Howarth and Sprague1978

Rainbow trout < 1 g FT, CuCl2 96-hr LC50 0.018 pH: 7.5; T: 9.8; Alk: 25; DO8.2 25 M, P Marr et al. 1995

Species Lifestage Test Type Toxicity Endpoint (mg Cu/L) Environmental Conditions Hardness Classification Reference

89

Sockeye salmon newly hatchedalevin frysmolt 4.8-5.5 g

96-hr LC50 0.1996-hr LC50 0.13 (0.1-0.2)96-hr LC50 0.1596-hr LC50 0.17-0.24

pH: 7.6 (7.55-7.8); T: 5.5-8.9; Alk: 62.5 (60.8-68.2); DO: Saturated

83.1 (82.9-84.4)

M, P Servizi and Martens1978

Steelhead AlevinSwim-upParrSmolt

FT 96-hr LC50 0.02896-hr LC50 0.01796-hr LC50 0.01896-hr LC50 0.029

pH: 7.1; T: 12.2+0.4; Alk: 22+2; DO: 10.2+0.3 23+1 M, P Chapman 1978

Steelhead Adult FT 96-hr LC50 0.057 pH: 7.57+0.06; T: 9.2 (7-11); Alk: 34+8;DO:11.4 (10.6-12)

42+12 M, P Chapman and Stevens1978

Steelhead trout juveniles FT 96-hr LC50 0.08 pH: 7.4-7.9; T: 12; Alk: 126; DO: 9.3-10 120 M, P Seim et al. 1984

Steelhead trout fry 2.36-3.01 g FT, CuCl2 96-hr LC50 0.002896-hr LC50 0.004296-hr LC50 0.0066

pH: 7; T: 15-16; Alk: 11;DO: >90% SatpH: 5.7; T: 15-16; Alk: 1.7; DO: >90% SatpH: 4.7; T: 15-16; Alk: -0.2; DO: >90% Sat

9.2 M, P Cusimano et al. 1986

Stone roller 60 mm FT, CuSO4 96-hr LC50 0.29 pH: 7.9-8.1; T: 23-25; Alk: 148-161 196-205 M, P Geckler et al. 1976

Stone loach 8.7-12.1 cm FT 4-d LC50 0.7 pH: 8.26+0.1; T: 11.9+0.2; DO: >96% sat 249+17 P Solbe and Cooper 1976

Striped shiner 55 mm55 mm

FT, CuSO4 96-hr LC50 0.7996-hr LC50 1.9

pH: 7.9-8.1; T: 23-25; Alk: 148-161 196-205 M, P Geckler et al. 1976

Zebrafish 0.2-0.6 g FT 96-hr LC50 0.149 pH: 8.0-8.3; T: 25; Alk: 200-255; DO: avg 75%sat

350-375 M, P Fogels and Sprague 1977

Zebrafish 0.29-0.69 mg FT,CuSO4

96-hr LC50 0.24 pH: 7.7; T: 24; Alk: 81; DO: 87% Sat 128 M, P Weinstein 1978

Secondary Studies

American eel S, CuNO3 96-hr LC50 6.096-hr LC50 6.4

pH: 8; T: 28; DO: 6.9pH: 7.8; T: 17; DO: 6.5

5553

N, S Rehwoldt et al. 1972/71

Arctic grayling fryalevin

juvenile

S, CuSO4 96-hr LC50 0.01096-hr LC50 0.024-0.13196-hr LC50 0.003-0.068

pH: 7.1-8; T: 12+1; Alk: 30.9 41.3 N, S Buhl and Hamilton 1990

Atlantic salmon 8.9-9.8 cm FT >96-hr ILL 0.025 T: 3.8-4.8 14 M, S Zitko et al. 1973

Atlantic salmon 2-3 yr old parr S 96-hr LC50 0.125 pH: 6.7-6.5; T: 18-21; Alk: 4 8-10 M, S Wilson 1972

Species Lifestage Test Type Toxicity Endpoint (mg Cu/L) Environmental Conditions Hardness Classification Reference

90

Atlantic salmon fry4-6cm

SR (1/d),CuSO4

21-d LC50 0.0421-d LC50 0.04

pH: 6.4; T: 10pH: 6.4; T: 14-16

14 N, S Grande 1967

Banded killifish <20 cm S, CuNO3 96-hr LC50 0.8496-hr LC50 0.86

pH: 8; T: 28; DO: 6.9pH: 7.8;T: 17; DO: 6.5

5553

N, S Rehwoldt et al. 1972/71

Bluegill 7 (5-11) cm S, CuSO4 48-hr LC50 348-hr LC50 748-hr LC50 44

pH: 8; T: 20; Alk: 43; DO: aeratedpH: 8.3; T: 20; Alk: 72; DO: aeratedpH: 8.6; T: 20; Alk: 144; DO: aerated

46101190

N, S Thurnbull et al. 1954

Bluegill 0.6 g S, CuSO4 96-hr LC50 0.4 pH: 8; T: 24; Alk: 48; DO: > 3 mg/L 52 N, S Inglis and Davis 1972

Bluegill 96-hr LC50 0.296-hr LC50 10

pH: 7.4pH: 8.3

20400

NR, S Tarzwell and Henderson1960

Bluegill 0.6 g S 96-hr LC50 1.02 pH: 8; T: 24; Alk: 48; DO: > 3 mg/L 365 N, S Inglis and Davis 1972

Bluegill 0.6 g S 96-hr LC50 0.68 pH: 8; T: 24; Alk: 48; DO: > 3 mg/L 209 N, S Inglis and Davis 1972

Bluegill 3.5-3.9 g S, CuCl2 96-hr LC50 1.25 T: 18+2; synth. dil. medium; DO: 5-9 N, S Cairns and Scheier 1968

Bluegill 1-2 g S 96-hr LC50 0.66 mg/L pH: 7.5 (7.1-7.4); T: 25; Alk: 18; DO: >4 mg/L,start 7.8 mg/L, not aerated

20 N, S Pickering and Henderson1966

Brook trout 5.2-11.2 cm FT,CuSO4

10-d LC50 0.05 T: 15 14 NR, S Sprague 1968

Brown trout yolk-sac fry SR (1/d), CuSO4

21-d LC50 0.04 pH: 6.4; T: 10; 14 N, S Grande 1967

Carp eggs SR (1/d) 3-d hatch NOEL 0.093-d hatch LOEL 0.5

pH: 7.3; T: 26 50 M-NR, S Hildebrand and Cushman1978

Carp 3.5-5.5 cm/g S, CuNO3 48-hr LC50 0.11548-hr LC50 0.27548-hr LC50 0.75

pH: 7.9-8.2; T: 20+1 50100300

N, S Peres and Pihan 1991b

Carp S, CuNO3 96-hr C50 0.896-hr LC50 0.81

pH: 8; T: 28; DO: 6.9pH: 7.8; T: 17; DO: 6.5

5553

N, S Rehwoldt et al. 1972/71

Carp 3.5-5.5 cm/g S, CuCl2 48-hr C50 0.11848r LC50 0.2948-hr LC50 0.751

pH: 7.5-8.2; T: 20+1; DO: >70% Sat 50100300

N, S Peres and Pihan 1991a

Catla catla 2.73+0.27 g S, CuSO4 96-hr C50 0.30 pH: 7.3-8; T: 29-30; DO: 7.08-8.0 460-465 N, S Ahmad and DattaMunshi 1987

Species Lifestage Test Type Toxicity Endpoint (mg Cu/L) Environmental Conditions Hardness Classification Reference

91

Channel catfish 3.9 g3.0 g2.3 g3.2 g

S, CuSO4 96-hr C50 0.05596-hr LC50 0.73196-hr LC50 0.95496-hr LC50 0.983

pH: 7.3;T: 17+0.5; Alk: 16; DO: >90% SatpH: 8.2; T: 17+0.5; Alk: 76; DO: >90% SatpH: 8.4; T: 17+0.5; Alk: 127; DO: >90% SatpH: 8.7;T: 17+0.5; Alk: 239; DO: >90% Sat

1683161287

N, S Straus and Tucker 1993

Channel catfish eggs4-d post-hatch

SR (2/d) 96-hr C50 7.5696-hr LC50 6.62

pH: 7.2-7.8; T: 20-24; DO: mod aerated, sat 100 (93-105) M-D, S Birge and Black 1979

Chinook salmon 0.66/0.87 g S, CuSO4 96-hr C50 0.058-0.054 pH: 7.0-8.3; T: 12+1; Alk: 88 211 N, S Hamilton and Buhl 1990

Clarias batraches 50 g SR 48-hr C50 41.5 pH: 7.5; T: 27; Alk: 90 170 N, S Mukherjee andBhattacharya 1977

Coho salmon 6 g SR (1/d) 96-hr C50 0.017 Cu2+ pH: 7.04-7.53; T: 13.5+1; Alk: 29+1; DO:8.7+0.7

33+4 M, S Buckley 1983

Coho salmon adult 2.7 kg FT 96-hr C50 0.046 pH: 7.29+0.09; T: 9.4 (8.7-11.1); Alk: 22+1;DO: 9.9+0.2

20+1 N, S Chapman and Stevens1978

Coho salmon alevinjuvenile

S, CuSO4 96-hr C50 0.019-0.02196-hr LC50 0.015-0.032

pH: 7.1-8; T: 12+1; Alk: 30.9 41.3 N, S Buhl and Hamilton 1990

Colisa fasciatus SR (1/d) 96-hr C50 0.9 pH: 7; T: 21; Alk: 118; DO: 7.3 mg/L 50 NR, S Pande and Shukla 1992

Common carp 1.4-2.6 g SR (1/d), CuSO4

96-hr C50 0.063 pH: 6.3 (6.25-6.35); T: 15 (13-16); Alk: 29 (25-33); DO: 6.8

19 (14-26) N, S Khangarot et al. 1983

Cyprinus carpio 78-195 mg2880-3630 mg

SR (1/d) 96-hr C50 0.11896-hr LC50 0.53

pH: 7.8-8.1; T: 17-22; Alk: 95-122; DO: 8-8.4 144-188 N, S Deshmukh and Marathe1980

Fathead minnow LC50 1.4 pH: 8.2; Alk: 360 400 NR, S Tarzwell and Henderson1960

Fathead minnow 2-6.9 cm S 96-hr TL50 0.6-0.98 (diss.Cu)

pH: 7.0-8.5; T: 4-27; Alk: 90-230 120-336 diss Cu M, S Brungs et al. 1976

Fathead minnow 1-2 g S 96-hr LC50 0.023-0.035mg/L96-hr LC50 1.14/1.76

pH: 7.5 (7.1-7.4) start 8.2; T: 25; Alk: 18; DO:>4 mg/L, start 7.8mg/L, not aeratedpH: 7.5 (7.1-7.4) start 8.2; T: 25; Alk:300; DO:>4 mg/L, start 7.8mg/L, not aerated

20360

N, S Pickering and Henderson 1966

Fathead Minnow adult FT 96-hr LC50 0.47 pH: 7.9; T: 18-22; Alk: 160 200 M?, S Mount 1968

Fathead minnow LC50 0.05 mg/L pH: 7.4; Alk: 18 20 NR, S Tarzwell and Henderson1960

Goldfish eggs/larvae SR (2/d), CuSO4

7-d LC50 5.2 pH: 7.2-7.8; T: 20-24; DO: moderate aeration 197 M-D, S Birge and Black 1979

Species Lifestage Test Type Toxicity Endpoint (mg Cu/L) Environmental Conditions Hardness Classification Reference

92

Goldfish 1-2 g S 96-hr LC50 0.036 pH: start 7.5 (7.1-7.4); T: 25; Alk: 18; DO: >4mg/L, start 7.8 mg/L, not aerated

20 N, S Pickering and Henderson1966

Guppies 0.186 g S, CuSO4 96-hr LC50 0.764 pH: 7.5 (7.4-7.8); T: 26.5 (24-27.5); Alk: 165(140-190); DO: 6.6 (5.8-7.5)

240 (210-280) N, S Khangarot et al. 1981

Guppies 0.1-0.2 g S 96-hr LC50 0.036 pH: start 7.5 (7.1-7.4); T: 25; Alk: 18; DO: >4mg/L, start 7.8 mg/L, not aerated

20 N, S Pickering and Henderson1966

Labeo rohita 2.46+0.27 g S, CuSO4 96-hr LC50 0.70 pH: 7.3-8; T: 29-30; DO: 7.08-8.0 460-465 N, S Ahmad and DattaMunshi 1987

Labeo rohita 40-50 mg3700-5600 mg

SR (1/d) 96-hr LC50 0.08796-hr LC50 0.295

pH: 7.8-8.1; T: 17-22; Alk: 95-122; DO: 8-8.4 144-188 N, S Deshmukh and Marathe1980

Largemouth bass eggs4-d post-hatch

SR (2/d) 96-hr LC50 6.9796-hr LC50 6.56

pH: 7.2-7.8; T: 20-24; DO: mod aerated, sat 100 (93-105) M-D, S Birge and Black 1979

Lebistes reticulatus 5.5-6.7 g43.7-74 g324-463 g

SR (1/d) 96-hr LC50 0.1696-hr LC50 0.27596-hr LC50 0.48

pH: 7.8-8.1; T: 17-22; Alk: 95-122; DO: 8-8.4 144-188 N, S Deshmukh and Marathe1980

Mosquitofish 0.68-0.81 g ?, CuNO3

CuSO4

Cu3OCl

96-hr LC50 0.04796-hr LC50 0.08096-hr LC50 0.046

pH: 7.52 (7.3-7.9); T: 27.1 (25.8-29); Alk: 52.3(45-79); DO: 5.5 (4.3-7.9)

31.5 (27-41) N, S Joshi and Rege 1980

Mystus vittatus SR (1/d) 96-hr LC50 0.7 pH: 7; T: 21; Alk: 118; DO: 7.3 mg/L 50 NR, S Pande and Shukla 1992

Ophicephaluspunctatus

60 g SR 48-hr LC50 70 pH: 7.5; T: 27; Alk: 90 170 N, S Mukherjee andBhattacharya 1977

Phoxinus laevis SR 48 hr,CuSO4

7-d ILL 0.5 mg/L pH: 7.6; T: 10; DO: aerated,sat 253 N, S Liepolt and Weber 1958

Pumpkinseed S, CuNO3 96-hr LC50 2.796-hr LC50 2.4

pH: 8; T: 28; DO: 6.9pH: 7.8;T: 17; DO: 6.5

5553

N, S Rehwoldt et al. 1972/71

Pumpkinseeds 1.2-2.2 g2.8-4.5 g5.3 g7.6 g

FT 96-hr LC50 1.24-1.396-hr LC50 1.66-1.6796-hr LC50 1.7496-hr LC50 1.94

pH: 7.2+0.2; T: 20+1; Alk: 85+2; DO: 79-89%sat.

125+5 M-NR, S Anderson and Spear1980a

Puntius conchorius 5+0.4 cm S 96-hr LC50 0.571 pH: 8.1+0.2; T: 13-22; Alk: 228+1.2; DO:8.6+0.1

310+0.112 N, S Pant et al. 1980

Rainbow trout 3.9-176 g FT 96-hr LC50 0.19-0.21 pH: 7.8+0.1; T: 10+1; Alk: 85+2; DO: 74-95%sat.

125+5 M-NR, S Anderson and Spear1980a

Species Lifestage Test Type Toxicity Endpoint (mg Cu/L) Environmental Conditions Hardness Classification Reference

93

Rainbow trout 0.36 g SR (1/d) CuSO4

48-hr LC50 0.1248-hr LC50 0.11

pH: 6.5; T: 10-11; Alk: 100pH: 7.5; T: 10-11; Alk: 100

250 NR, S Shaw and Brown 1974

Rainbow trout 7.6 cm SR 72-hr LC50 1.107-d LC50 0.044

T: 16-17; DO: aeratedT: 17-18; DO: aerated

32015-20

N, S Lloyd 1961b

Rainbow trout 12.5 cm SR, CuSO4 48-hr LC50 0.27 pH: 7.8; T: 17; Alk: 250; DO: air saturation 320 N, S Herbert and Van Dyke1964

Rainbow trout 51-76 mm FT 96-hr LC50 0.253 pH: 6.4-8.3; Alk: 82-132; DO: 4.8-9 field study, N,S

Hale 1977

Rainbow trout ILL 0.037ILL 0.050ILL 0.09ILL 0.50

142040320

NR, S Lloyd and Herbert 1962

Rainbow trout yolk-sac fry SR (1/d), CuSO4

21-d LC50 0.04 pH: 6.4; T: 10 14 N, S Grande 1967

Rainbow trout 10 cm SR (1/d), CuSO4

7-d inc. lethal 0.50ILL 0.07-0.13

pH: 7.6-7.0; T: 10+0.2; DO: aerated, sat 25320

N, S Liepolt and Weber 1958

Rainbow trout alevinjuvenile

S, CuSO4 96-hr LC50 0.03696-hr LC50 0.014

pH: 7.1-8; T: 12+1; Alk: 30.9 41.3 N, S Buhl and Hamilton 1990

Rainbow trout embryo/alevin 4-d post hatch

SR (2/d), CuSO4

96-hr LC50 0.11 pH: 7.2-7.8; T: 12-13; DO: moderate aeration, near sat.

100 (93-105) M-D, S Birge and Black 1979

Rainbow trout yolk-sac fry SR (1/d), CuSO4

21-d LC50 0.04 pH: 6.4; T: 10 14 N, S Grande 1967

Rasboradeniconiusneilger

3.8 g S, CuSO4 96-hr LC50 0.203 pH: 7.5 (7.4-7.8); T: 26.5 (24-27.5); Alk: 165(140-190); DO: 6.6 (5.8-7.5)

240 (210-280) N, S Khangarot et al. 1981

Steelhead trout 8.7+1.4cm FT 96-hr LC50 0.020-0.025 T: 12; DO: 10 30-66 M, S Knittel 1980

Striped bass 63 d old S, CuSO4 96-hr LC50 0.196-hr LC50 0.27

pH: 8.1;T: 20+2; Alk: 30pH: 7.9; T: 20+2; Alk: 262

40285

N, S Palawski et al. 1985

Striped bass S, CuNO3 96-hr LC50 4.096-hr LC50 4.3

pH: 8; T: 28; DO: 6.9pH: 7.8; T:17; DO: 6.5

5553

N, S Rehwoldt et al. 1972/71

Striped bass 2.7 g S, CuSO4 96-hr LC50 0.62 pH: 8.2; T: 21; Alk: 35; DO: 7.8 64 N, S Wellborn 1969

White perch S, CuNO3 96-hr LC50 6.496-hr LC50 6.2

pH: 8; T: 28; DO: 6.9pH: 7.8; T:17; DO: 6.5

5553

N, S Rehwoldt et al. 1972/71

Species Lifestage Test Type Toxicity Endpoint (mg Cu/L) Environmental Conditions Hardness Classification Reference

Unsuitable/Unclassified studies

Blue gourami adult S 96-hr LC50 0.091 pH: 7.4; T: 26-28; DO: 10 N, U Roales and Perlmutter1974

Bluegill 7 (5-11) cm S, CuSO4 48-hr LC50 49 pH: 6.9-7.5; T: 20; Alk: 33-81; DO: aerated 84-163 N, U Thurnbull et al. 1954

Coho salmon 56-d 96-hr LC50 0.06 Hedtke et al. 1978

Coho salmon adult 96-hr LC50 0.046 pH: 7.2-7.4; T: 8.7-11.4 20 NR,U Chapman 1973

Coho salmon 130-d 96-hr LC50 0.166 Hedtke et al. 1978

Coho salmon 151-d 96-hr LC50 0.212 Hedtke et al. 1978

Rainbow trout 96-hr LC50 0.85 250 Ministry of Technology1973

Rainbow trout 0.3 NR, U Wong 1989

Steelhead trout juvenile 96-hr LC50 0.02 pH: 7.4; T: 12 20-25 NR, U Chapman 1973

Tench 0.08-0.15 pH: 7.7; T: 16 100 Haider 1966

Zebrafish LC50 0.24 pH: 7.7; T: 25; Alk: 81 130 Anderson et al. 1978

Zebrafish LC50 0.67 pH: 7.8T: 25Alk: 81 300 Anderson et al. 1978

Legend: Test Type: S=Static, SR=Static Replacement, FT=Flow-Through;Toxicity Endpoint: T=expressed as total copper, diss= expressed as dissolved copper, Cu2+= expressed as cupric ionEnvironmental Conditions: T= Water Temperature oC, Alk=Alkalinity in mg/L CaCO3, DO=Dissolved Oxygen in mg/L unless otherwise

stated; Hard=Water Hardness in mg/L CaCO3;Classification: M=Measured Concentrations, M-D= Measured Concentration, but significant decrease over test duration,

N=Nominal, NR=Not Reported; C=Calculated; RP=ReplicatedP=Primary, S=Secondary, U=Unsuitable.

95

Appendix 4. Acute copper toxicity data on freshwater invertebrates.

Species Lifestage Test Type Toxicity Endpoint (mg Cu/L) Environmental Conditions Hardness Classification Reference

Primary Studies

Asellus meridianus 4-6 mm SR 48-hr LC50 1.2-2.5 T:20+3; HMSO med. 25 M, P Brown 1976

Cambarus robustus intermoult adult S 24-96-hr LC50 0.83-3.48 pH: 7.0-6.1; T: 25+1; Alk: 10-12; DO: 7.5-8.3 10-12 M, P Taylor et al. 1995

Campelomadecisum

11-27 mm FT, CuSO4 96-hr LC50 1.7 mg/L pH: 7.7; T: 15; Alk: 42-43; DO: 9.5-9.7 44-45 M, P Arthur and Leonard 1970

Caridina sp. juv. FT 96-hr LC50 0.003-0.004 pH: 5.85-6.15; T: 27+0.5; Alk: 5 25-30 M, PAustralia

Williams et al. 1991

Ceriodaphnia dubia <12 hr

adult 5-6d

S 48-hr LC50 0.019-0.02748-hr LC50 0.024-0.05348-hr LC50 0.079-0.09948-hr LC50 0.079-0.12748-hr LC50 0.063-0.071

pH: 7.72+0.2; T: 25+1;Alk: 39.7 pH: 8.15+0.06; T: 25+1; Alk: 69.6pH: 8.31+0.03; T: 25+1;Alk: 140.1 pH: 8.31+0.03; T: 25+1; Alk: 140.1 pH: 8.15+0.06; T: 25+1; Alk: 69.6

459417917994.1

M, P Belanger et al. 1989

Ceriodaphnia dubia <24 hr S, CuNO3 48-hr LC50 0.066 pH: 8.2 (8-8.5); T: 25+2; Alk: 97+9.3; DO:>70%

100+7.9 M, P Spehar and Fiandt 1986

Ceriodaphnia dubia <4 hr S, CuCl2 48-hr LC50 0.01948-hr LC50 0.020

pH: 7.7;T: <22-28; Alk: 55; DO: 7.6pH: 7.5; T: <22-28; Alk: 38; DO: 7.6

5236

M, 2RP, P Carlson et al. 1986

Ceriodaphnia dubia neonate SR (1/d) 48-hr LC50 0.05648-hr LC50 0.08448-hr LC50 0.093

pH: 6; T: 25; Alk: 144.3+8.4pH: 8; T: 25; Alk: 144.3+8.4pH: 9; T: 25; Alk: 144.3+8.4

182.0+10.1 M, P Belanger and Cherry 1990

Ceriodaphnia dubia neonate SR (1/d) 48-hr LC50 0.01448-hr LC50 0.02848-hr LC50 0.031

pH: 6; T: 25; Alk: 74.2+6.1pH: 8; T: 25; Alk: 74.2+6.1pH: 9; T: 25; Alk: 74.2+6.1

97.6+3.9 M, P Belanger and Cherry 1990

Ceriodaphnia dubia neonate SR (1/d) 48-hr LC50 0.05248-hr LC50 0.07648-hr LC50 0.091

pH: 6; T: 25; Alk: 121.9+8.2pH: 8; T: 25; Alk: 121.9+8.2pH: 9; T: 25; Alk: 121.9+8.2

113.6+4.2 M, P Belanger and Cherry 1990

Chironomusriparius

2nd instar SR (1/d) 48-hr LC50 0.7 pH: 6.8-7.2; T: 20; DO: >80% Sat 151+9 M, P Taylor et al. 1991

Chironomusriparius

2nd instar S 48-hr LC50 1.17 pH: M; T: 20; Alk: 50-60; DO: 80% Sat 100-120 M, P Dobbs et al. 1994

Chironomus tentans 2nd instar FT, CuCl2 96-hr LC50 0.773 pH: 7.4; T: 19.5; Alk: 60-67 84 M, P Nebeker et al. 1984b

Species Lifestage Test Type Toxicity Endpoint (mg Cu/L) Environmental Conditions Hardness Classification Reference

96

Chironomus tentans

3rd instar FT, CuCl2 96-hr LC50 1.446 pH: 7.4; T: 19.5; Alk: 60-67 84 M, P Nebeker et al. 1984b

Chironomus tentans

4th instar FT, CuCl2 96-hr LC50 1.69 pH: 7.4; T: 19.5; Alk: 60-67 84 M, P Nebeker et al. 1984b

Daphnia magna FT 48-hr LC50 0.02 pH: 7.2-7.6; DO: 8.5 120-130 M, P Bishop and Perry 1981

Daphnia magna <24 hr SR (1/2d),CuCl2

96-hr LC50 0.13 Alk: 82 85 M, P Blaylock et al. 1985

Daphnia magna <24 hr S, CuO 48-hr LC50 0.026 pH: 7.1-7.9; T: 22+1; DO: 7.9+0.3 143+9.8 M, P Lewis 1983

Daphnia pulex 24 hr + 10 hr CuNO3 48-hr LC50 0.003 pH: 7.5-8.5; T: 20+2; Alk: 30-45 45-60 M, P Jop et al. 1993

Daphnia pulex <24 hr S 48-hr LC50 0.037 pH: M; T: 20; Alk: 50-60; DO: 80% Sat 100-120 M, P Dobbs et al. 1994

Daphnia pulex <24 hr S 72-hr LC50 0.025972-hr LC50 0.02872-hr LC50 0.0136

pH: 8.4-8.7; T: 20+1; Alk: 115 57.5115230

M, P Winner 1985

Ephemerellagrandis

FT, CuSO4 14-d LC50 0.18-0.2 pH: 7.0-7.2 (one exp. 6.3); T: 3-9 (<0.5); Alk:30-70 (<10); DO: 7-12 (<0.5)

30-70 (<10) M, P Nehring 1976

Gammarus pulex 3-5 mm SR (1/d) 48-hr LC50 0.037 pH: 6.8-7.2; T: 12+1; DO: >80% Sat 151+9 M, P Taylor et al. 1991

Gammaruspseudolimneaus

adult FT, CuSO4 96-hr LC50 0.02 pH: 7.7; T: 15; Alk: 42-43; DO: 9.5-9.7 44-45 M, P Arthur and Leonard 1970

Hyalella azteca 7-14d FT, CuSO4 10-d LC50 0.031 pH: 7.5-8.3; T: 22.1-22.3; Alk: 45-46; DO: 6.9-7.9 44-47 M, P West et al. 1993

Hydropsyche betteni S 96-hr LC50 >6414-d LC50 32

pH: 7.25; T: 18.5; Alk: 40; DO: aeration, 8mg/L 44-46 M, P Warnick and Bell 1969

Isonychia bicolor 6-8 instar S 48-hr LC50 0.223 pH: M; T: 20; Alk: 50-60; DO: 80% Sat 100-120 M, P Dobbs et al. 1994

Keratella cochlearis CuCl2 LC50 0.101 pH: 8.3; T: 20; Alk: 103; 180 M, P Borgmann and Ralph 1984

Lumbriculus variegatus 7 mg FT, CuSO4 10-d LC50 0.035 pH: 7.5-8.3; T: 22.1-22.3; Alk: 45-46; DO: 6.9-7.9 44-47 M, P West et al. 1993

Orconectes sp. 30-40 mm S 48-hr LC50 2.37 pH: M; T: 20; Alk: 50-60; DO: 80% Sat 100-120 M, P Dobbs et al. 1994

Paratya australiensis adult FT, CuSO4 96-hr LC50 0.034(tot)/0.016(Cu2+)

pH: 6.96 (6.49-7.26); T: 15; Alk: 9.1 (2.6-; 12.2);DO: aerated

17.0 (11.5-23.1) M, P (2RP) Daly et al. 1990a

Physa integra 4-7 mm FT, CuSO4 96-hr LC50 0.039 pH: 7.7; T: 15; Alk: 42-43; DO: 9.5-9.7 44-45 M, P Arthur and Leonard 1970

Physella sp. <10mm t S 48-hr LC50 0.109 pH: M; T: 20; Alk: 50-60; DO: 80% Sat 100-120 M, P Dobbs et al. 1994

Species Lifestage Test Type Toxicity Endpoint (mg Cu/L) Environmental Conditions Hardness Classification Reference

97

Pteronarcys californica

FT, CuSO4 14-d LC50 10.1-13.9 pH: 7.0-7.2 (one exp. 6.3); T: 3-9 (<0.5); Alk: 30-70(<10); DO: 7-12 (<0.5)

30-70 (<10) M, P Nehring 1976

Scapholeberis sp. adult S, CuCl2 48-hr LC50 0.018 pH: 7.7; T: <22-28; Alk: 55; DO: 7.9-8.1 52 M, P Carlson et al. 1986

Stenonema sp. 6-8 instar S 48-hr LC50 0.453 pH: M; T: 20; Alk: 50-60; DO: 80% Sat 100-120 M, P Dobbs et al. 1994

Secondary Studies

Acroneuria lycorias

S 96-hr LC50 8.3 pH: 7.25; T: 18.5; Alk: 40-54; DO: aeration 8 mg/L 44-40 M-D,S Warnick and Bell 1969

Amnicola sp. adultseggs

S, CuNO3 96-hr LC50 0.996-hr LC50 9.3

pH: 7.6; T: 17; DO: 6.2 50 N, S Rehwoldt et al. 1973

Biomphalaria glabrata

adult FT,SSW3 24-hr LC50 0.41/0.28 (tot/Cu2+)24-hr LC50 0.11/0.052 (tot/Cu2+)24-hr LC50 0.56/0.022 (tot/Cu2+)

pH: 6; T: 25pH: 7; T: 25pH: 8; T: 25

M, S O'Sullivan et al. 1989

Brachionus calyciflorus

<2 hr S, CuSO4 24-hr LC50 0.037 pH: 7.5; T: 25; synthetic freshwater N, S Porta and Ronco 1993

Brachionus rubens neonate S, CuSO4 24-hr LC50 0.019 pH: 7.4-7.8: T: 25; Alk: 60-70 80-100 NR, S Snell and Persoone 1989

Brachionus calyciflorus

neonates CuSO4 24-hr LC50 0.76 pH: 7.4-7.8; T: 25; Alk: 60-70 80-100 N, S Ferrando et al. 1992

Caddis fly S, CuNO3 96-hr LC50 6.2 pH: 7.6; T: 17; DO: 6.2 50 N, S Rehwoldt et al. 1973

Ceriodaphnia reticulata

<4 hr S 48.hr LC50 0.017 pH: 7.2-7.4; T: 25; Alk: 43-45; DO: >5 45 N, S Mount and Norberg 1984

Ceriodaphnia reticulata

<24 hr S, CuCl2 48-hr EC50 0.023 pH: 8.0+0.3; T: 23+1; Alk: 230+10 240+10 N, S, 2RP Elnabarawy et al. 1986

Chironomus tentans S, CuSO4 48-hr EC50 0.327 pH: 6.3 (6.1-6.6); T: 14 (13-17); Alk: 25 (20-33);DO: 6.5 (5.5-8.0)

25 (18-35) N, S Khangarot and Ray 1989

Chironomus decorus 4th instar S, CuSO4 48-hr LC50 0.74 pH: 7.3-7.8; T: 20; Alk: 30-35; DO: aerated 40-48 M, S Kosalwat and Knight 1987

Chironomus tentans 2nd instar (12 d) FT, CuSO4 10-d LC50 0.059 pH: 7.5-8.3; T: 22.1-22.3; Alk: 45-46; DO: 6.9-7.9 44-47 control?, S West et al. 1993

Chironomus tentans 1st instar (12 hr) S, CuSO4 immobility96-hr EC50 0.016796-hr EC50 0.036596-hr EC50 0.0982

DO: pre-aeratedpH: 7.6+0.1; T: 21+1; Alk: 32.4+1.1; pH: 7.8+0.1; T: 21+1; Alk: 70.9+1; pH: 8.1+0.1; T: 21+1; Alk: 111.3+1.5;

42.7+0.7109.6+1.2172.3+1.9

N, S Gauss et al. 1985

Species Lifestage Test Type Toxicity Endpoint (mg Cu/L) Environmental Conditions Hardness Classification Reference

98

Chironomus thummi SR?, CuSO4 7-d lethal threshold 0.5 pH: 7.6; T: 10+0.2; DO: aerated 253 N, S Liepolt and Weber 1958

Chironomus sp. S, CuNO3 96-hr LC50 0.03 pH: 7.6; T: 17; DO: 6.2 50 N, S Rehwoldt et al. 1973

Chironomus tentans 1st instar SR (2/d), CuCl2 96-hr LC50 0.298 pH: 7.4; T: 20; Alk: 60-67; DO: 4 71 M, fed, S Nebeker et al. 1984b

Crayfish intermoult adult 30-35 mm

FT, CuSO4 96-hr LC50 3 mg/L pH: 7.8-8.1; T: 20 100-125 N, S Hubschman 1967

Cyclops abyssorum 0.62 mm S 48-hr LC50 2.5 pH: 7.2; T: 10+0.5; Alk: 0.58 meq/L; DO: no sign.change

N, S Baudouin and Scoppa 1974

Damsel fly S, CuNO3 96-hr LC50 4.6 pH: 7.6; T: 17; DO: 6.2 50 N, S Rehwoldt et al. 1973

Daphnia pulex CuCl2 EC50 0.064 Porcella EPA med. N, S Miller et al. 1985

Daphnia magna 12-hr S 72-hr LC50 0.086 pH: 8.2-9.5; T: 20; Alk: 100-119; DO: 8.7-11.4 130-160 N, S Winner and Farrell 1976

Daphnia magna 0-24 hr S Immob 48-hr EC50 0.032 pH: 8.4; T: 20; artificial med. M,S Borgmann and Ralph 1983

Daphnia similis CuSO4 96-hr LC50 0.041 T: 28-30 N, S Soundrapandian and Venkataraman1990

Daphnia magna <8 hr S, CuCl2 48-hr Threshold 0.027 T: 25 N, S Anderson 1948

Daphnia magna 6-24 hr S, CuSO4 48-hr LC50 0.022 pH: 7.2; T: 20; Alk: 21.7; DO: aerated 10 N, S Hickey and Vickers 1992

Daphnia magna 6-24 hr CuSO4 48-hr LC50 0.0065 pH: 8-8.1; T: 20.5-21; DO: Sat 250 N, S Dave 1984

Daphnia magna 12 hr S, CuSO4 72-hr LC50 0.081-0.089 mg/L pH: 7.2-9.5; T: 20+1; Alk: 100-118; DO: 7.3-11.4 N, fed, S Winner et al. 1977

Daphnia magna <24 hr S 48-hr LC50 0.036 T: 20; artificial medium M, S Borgmann and Charlton 1984

Daphnia ambigua 12 hr S 72-hr LC50 0.0677 pH: 8.2-9.5; T: 20; Alk: 100-119; DO: 8.7-11.4 130-160 N, S Winner and Farrell 1976

Daphnia parvula 12 hr S 72-hr LC50 0.072 pH: 8.2-9.5; T: 20; Alk: 100-119; DO: 8.7-11.4 130-160 N, S Winner and Farrell 1976

Daphnia magna <24 hr S, CuCl2 48-hr EC50 0.041 pH: 8.0+0.3; T: 23+1; Alk: 230+10 240+10 N, S, 2RP Elnabarawy et al. 1986

Daphnia magna <24 hr S 48-hr EC50 0.007 pH: 6.5; T: 20+0.5; synthetic medium N, S Oikari et al. 1992

Daphnia magna S, CuSO4 48-hr EC50 0.093 pH: 7.6 (7.4-7.8); T: 13 (11.5-14.5); Alk: 400 (390-415); DO: 5.6 (5.2-6.5)

240 (235-260) N, S Khangarot and Ray 1987b

Daphnia magna S, CuSO4 24-hr LC50 0.53648-hr LC50 0.093

pH: 7.6 (7.4-7.7); T: 13; Alk: 400; DO: 5.6 (5.2-6.5) 240 N, S Khangarot et al. 1987

Daphnia magna 6-24 hr SR (3/wk) 24-hr LC50 0.3448-hr LC50 0.37

pH: 7.4-7.8; T: 25; Alk: 60-70 80-100 N, S Ferrando et al. 1992

Species Lifestage Test Type Toxicity Endpoint (mg Cu/L) Environmental Conditions Hardness Classification Reference

99

Daphnia magna 12 hr SR (7d), CuCl2 3-wk LC50 0.044 48-hr LC50 with food 0.06without food 0.0098

pH: 7.4-8.2; T: 18; Alk: 42; DO: near saturation 45 N, S Biesinger and Christensen 1972

Daphnia magna <24-hr SR 48-hr LC50 0.073 T: 20 S (info testsolution)

Arambasic et al. 1995

Daphnia pulex <24 hr S, CuCl2 48-hr EC50 0.031 pH: 8.0+0.3; T: 23+1; Alk: 230+10 240+10 N, S, 2RP Elnabarawy et al. 1986

Daphnia pulex 12 hr S 72-hr LC50 0.086 pH: 8.2-9.5; T: 20; Alk: 100-119: DO: 8.7-11.4 130-160 N, S Winner and Farrell 1976

Daphnia hyalina 1.27 mm S 48-hr LC50 0.005 pH: 7.2; T: 10+0.5; Alk: 0.58 meq/L; DO: nosignificant change

N, S Baudouin and Scoppa 1974

Deleatidium sp. 3-6mm S, CuSO4 96-hr LC50 0.039 pH: 7.2; T: 15; Alk: 21.7; DO: aerated 10 N, S Hickey and Vickers 1992

Ephemerella subvaria S 48-hr LC50 0.32 pH: 7.25; T: 18.5; Alk: 40-42; DO: aeration 8 mg/L 44-40 M-D, S Warnick and Bell 1969

Eudiaptomus padanus 0.43 mm S 48-hr LC50 0.50 pH: 7.2; T: 10+0.5; Alk: 0.58 meq/L; DO: no sign.change

N, S Baudouin and Scoppa 1974

Gammarus sp. S, CuNO3 96-hr LC50 0.91 pH: 7.6; T: 17; DO: 6.2 50 N, S Rehwoldt et al. 1973

Gammarus fasciatus S, CuSO4 48-hr LC50 0.19 pH: 7.75; T: 11.8; DO: aerated, 12 mg/L 206 mg/L N, S Judy 1979

Gammarus pulex SR (1/d), CuCl2 96-hr LC50 0.10996-hr LC50 0.021

pH: 8.33; T: 7; DO: 10.9 pH: 8.33; T: 15; DO: 10.9

249104

N, S Stephenson 1983

Gammarus pulex SRCuSO4

7-d lethal threshold 0.25-0.5 pH: 7.6; T: 10+0.2; DO: aerated 253 N, S Liepolt and Weber 1958

Goniobasis livescens S, CuSO4 96-hr LC50 0.39 pH: 8.5; T: 15; DO: 5-6 154 M-D, S Paulson et al. 1983

Goniobasis livescens CuSO4 48-hr LC50 0.86 pH: 8-8.6 (dropped 0.3-1); T: 23.5+2.5; Alk: 115;DO: 8-9 dropped to 6-8 mg/L

137-171 NR, S Cairns et al. 1976

Gyraulus circumstriatus

S 96-hr LC50 0.108 pH: 7.85; T: 21.1 100 N, S Wurtz and Bridges 1961

Heptagenia lateralis SR?, CuSO4 7-d lethal threshold 0.5 pH: 7.6; T: 10+0.2; DO: aerated 253 N, S Liepolt and Weber 1958

Hydrobia jenkinsi 3-4.5mm S 72-hr NOEL 072-hr LC100 0.01

pH: 7.4 N, S Brown 1980

Limnodrillus hoffmeisteri

S 96-hr LC50 0.102 pH: 7.85; T: 22 100 N, S Wurtz and Bridges 1961

Species Lifestage Test Type Toxicity Endpoint (mg Cu/L) Environmental Conditions Hardness Classification Reference

100

Lophopodella carteri

2-3 d S 96-hr LC50 0.51 pH: 7.4-8.0; T: 24-25; DO: 7.5-8.3 190-220 N, S Pardue and Wood 1980

Lumbriculus variegatus

CuSO4 96-hr LC50 0.15 pH: 7.8; T: 20+1; Alk: 30; DO: >40% Sat 30 N, S Baily and Liu 1980

Lymnaea accuminata 0.55 g S, CuSO4 96-hr LC50 0.034 pH: 7.5 (7.4-7.8); T: 26.5 (24-27.5); Alk: 165(140-190); DO: 6.6 (5.8-7.5)

240 (210-280) N, S Khangarot et al. 1981

Lymnaea emarginata CuSO4 48-hr LC50 0.3 pH: 8-8.6 (dropped 0.3-1); T: 23.5+2.5; Alk: 115;DO: 8-9 dropped to 6-8 mg/L

137-171 NR, S Cairns et al. 1976

Nais sp. S, CuNO3 96-hr LC50 0.09 pH: 7.6; T: 17; DO: 6.2 50 N, S Rehwoldt et al. 1973

Paratya australiensis SR (1/2d) 96-hr LC50 0.034 pH: 6.96; T: 15; Alk: 9.1; DO: aerated 17 N, S Daly et al. 1990a

Pectinatella magnifica 2-3 d S 96-hr LC50 0.14 pH: 7.4-8.0; T: 24-25; DO: 7.5-8.3 190-220 N, S Pardue and Wood 1980

Philodina acuticornis S, CuSO4 96-hr LC50 0.7 96-hr LC50 1.1

pH: 7.4-7.9; T: 20+2; Alk: 24pH: 7.4-7.8; T: 20+2; Alk: 54-67

25 mg/L81 mg/L

N, S Buikema et al. 1974

Physa heterostropha S, CuSO4 96-hr LC50 0.069 pH: 7.85; T: 21.1 100 N, S Wurtz and Bridges 1961

Physa & Stagnicola sp.

S, CuSO4 96-hr LC50 0.083 mg/L pH: 7.8; T: 22 131 N, S Howard et al. 1964

Plumatella emarginata 2-3 d S 96-hr LC50 0.14 pH: 7.4-8.0; T: 24-25; DO: 7.5-8.3 190-220 N, S Pardue and Wood 1980

Plumatella casmiana CuSO4 96-hr LC50 0.50-1.00 T: 23-24 N, S Bushnell 1974

Polypedilum nubifer 1st instar2-3rd instar4th instar

S, CuSO4 48-hr LC50 0.0548-hr LC50 0.6348-hr LC50 4.3

T: 22+1 soft water N, S Hatakeyama 1988

Tubifex tubifex - SR (24-hr), CuSO4 48-hr LC50 0.2148-hr LC50 0.89

pH: 7.2&6.85; T: 20; Alk: 7.2&22.5pH: 7.32; T: 20; Alk: 234

34.2 mg/L261 mg/L

N, S Brkovic-Popovic and Popovic 1977

Tubifex rivulorum SR, CuSO4 7-d lethal threshold 0.25-0.5 pH: 7.6; T: 10+0.2; DO: aerated 253 N, S Liepolt and Weber 1958

Viviparous bengalensis

1.916 g S, CuSO4 96-hr LC50 0.088 pH: 7.5 (7.4-7.8); T: 26.5 (24-27.5); Alk: 165 (140-190); DO: 6.6 (5.8-7.5)

240 (210-280) N, S Khangarot et al. 1981

Unsuitable Studies

Daphnia magna <8 hr old S 32-hr LC50 0.038 T: 25 U Anderson 1944

Daphnia 0.16 no info, U Wong 1989

Species Lifestage Test Type Toxicity Endpoint (mg Cu/L) Environmental Conditions Hardness Classification Reference

101

Daphnia pulex LC50 0.05-0.1 T: 20 215 Ivekovic 1932

Lamellidens marginalis

5-6 cm SR (1/d) 96-hr LC50 5.0 U Raj and Hameed 1991

Moina irrasa <24 hr S 48-96-hr LC50 6.03-3.348-96-hr LC50 8.09-6.0648-96-hr LC50 12.61-7.43

pH: 5; T: 20; distilledpH: 6.5; T: 20; distilledpH: 8; T: 20; distilled

N, U Zou and Bu 1994

Physa heterostropha 3-6mm3-6mm12-15mm

CuSO4 48-hr LC50 0.01996-hr LC50 0.01648/96-hr LC50 0.013 48/96-hr LC50 0.069

pH: 7.3; T: 22pH: 7.3; T: 22pH: 7.8; T: 22pH: 7.3; T: 22

2020100100

NR, UNR, UNR, UNR, U

Wurtz 1962

Physa & Stagnicola sp. S, CuSO4 96-hr LC50 0.035 pH: 7.4; T: 22 10 N, U Howard et al. 1964

Viviparus bengalensis 2.8-3.5 g S, CuSO4 96-hr LC50 2.4 pH: 7.45; T: 30+1; Alk: 18; DO: 6.76 52 N, U Seth et al. 1990

Legend: Test Type: S=Static, SR=Static Replacement, FT=Flow-Through;Toxicity Endpoint: T=expressed as total copper, diss= expressed as dissolved copper, Cu2+= expressed as cupric ionEnvironmental Conditions: T= Water Temperature oC, Alk=Alkalinity in mg/L CaCO3, DO=Dissolved Oxygen in mg/L unless otherwise stated;

Hard=Water Hardness in mg/L CaCO3;Classification: M=Measured Concentrations, M-D= Measured Concentration, but significant decrease over test duration,

N=Nominal, NR=Not Reported; C=Calculated; RP=ReplicatedP=Primary, S=Secondary, U=Unsuitable.

102

Appendix 5. Computations for the acute Alberta copper guideline.

A. Computational layout for ANCOVA: notations as used in Box 14.10 (Sokal and Rohlf1981).

Ceriodaphnia Chinook salmon Fathead minnow Rainbow trout

df 16 6 25 29

SSY 6.877687 4.601996 27.11695 72.34533

SSX 3.894139 8.748168 11.6754 50.75116

SSXY 3.97945 6.023679 10.71988 52.77864

bY?X 1.021908 0.688565 0.91816 1.03995

explained SS 4.06663 4.147693 9.842564 54.88713

df 15 5 24 28

SSY?X 2.811057 0.454304 17.27438 17.4582

MSY?X 0.187404 0.090861 0.719766 0.623507

Pooled regression slope bwithin = 73.50165/75.06886 = 0.979123Explained variance SS (within) = (73.50165)2/75.06886 = 71.96716Unexplained variance Ed2

Y?Xwithin = 110.942-7196716 = 38.9748s2

Y?Xwithin = 38.9748/75 = 0.519664Fs = 71.96716/0.519664 = 138.4879; highly significant

EEd2Y?X = 37.99795

s2Y?X = 37.99795/72 = 0.527749

SSamong b's = 0.976849MSamong b's = 0.976849/3 = 0.325616Fs = 0.325616/0.527749 = 0.616991; not significant

103

B Computations Species Mean Acute Values (SMAV) and Genus Mean Acute Values(GMAV). LC50 values are in mg/L and water hardness in mg/L CaCO3.

LC50 hardness set Z=100Y

adj LC50 SMAV GMAV

2-linedsalamander

Dobbs et al.1994

1.12 110 0.020008 1.02021 1.02021 1.02021

asellus Brown 1976 1.73 25 1.905474 6.722594 6.722594 6.722594cambarus Taylor et al.

19951.699529 11 2.691545 14.75445 14.75445 14.75445

campeloma Arthur andLeonard1970

1.7 44 1.334469 3.797979 3.797979 3.797979

caridina Williams etal. 1991

0.003 27 -4.52714 0.010812 0.010812 0.010812

ceriodaphnia Belanger etal. 1989

0.021354 45 -3.06467 0.046669

ceriodaphnia Belanger etal. 1989

0.035665 94 -3.273 0.037893

ceriodaphnia Belanger etal. 1989

0.088436 179 -2.99553 0.05001

ceriodaphnia Belanger etal. 1989

0.100165 179 -2.871 0.056642

ceriodaphnia Belanger etal. 1989

0.06688 94.1 -2.64531 0.070984

ceriodaphnia Belangerand Cherry1990

0.056 182 -3.46874 0.031156

ceriodaphnia Belangerand Cherry1990

0.084 182 -3.06327 0.046734

ceriodaphnia Belangerand Cherry1990

0.093 182 -2.96149 0.051742

ceriodaphnia Belangerand Cherry1990

0.014 97.6 -4.24491 0.014337

ceriodaphnia Belangerand Cherry1990

0.028 97.6 -3.55177 0.028674

ceriodaphnia Belangerand Cherry1990

0.031 97.6 -3.44998 0.031746

ceriodaphnia Belangerand Cherry1990

0.052 113.6 -3.08136 0.045897

LC50 hardness set Z=100Y

adj LC50 SMAV GMAV

104

ceriodaphnia Belangerand Cherry1990

0.076 113.6 -2.70187 0.067080.04466 0.04466

ceriodaphnia Belangerand Cherry1990

0.091 113.6 -2.52175 0.080319

ceriodaphnia Spehar andFiandt 1986

0.066 100 -2.7181 0.066

ceriodaphnia Carlson etal. 1986

0.019 52 -3.32304 0.036043

ceriodaphnia Carlson etal. 1986

0.02 36 -2.9117 0.054383

chironomus rip. Taylor et al.1995

0.7 151 -0.76018 0.4675820.705923 0.804523

chironomus rip. Dobbs et al.1994

1.17 110 0.063683 1.065755

chironomus ten. Nebeker etal. 1984a

0.773 84 -0.08676 0.916895 0.916895

daphnia magna Bishop andPerry 1981

0.02 125 -4.13051 0.0160750.035538 0.024277

daphnia magna Blaylock etal. 1985

0.13 85 -1.88109 0.152423

daphnia magna Lewis1983 0.026 143 -3.99987 0.018318Daphnia pulex Jop et al.

19930.003 52 -5.16887 0.005691

0.016585Daphnia pulex Dobbs et al.

19940.037 110 -3.39016 0.033703

Daphnia pulex Winner1985

0.0259 57.5 -3.11168 0.044526

Daphnia pulex Winner1985

0.028 115 -3.71239 0.024419

Daphnia pulex Winner1985

0.0136 230 -5.11321 0.006017

Ephemerella gr. Nehring1976

0.19 46 -0.90041 0.406401 0.406401 0.406401

G a m m a r u spulex

Taylor et al.1991

0.037 151 -3.70034 0.024715 0.0247150.033231

G. pseudolimn. Arthur andLeonard1970

0.02 44 -3.10818 0.044682 0.044682

Hyallella azteca West et al.1993

0.031 45 -2.69193 0.06775 0.06775 0.06775

Hydropsyche b. Warnick andBell 1969

64 45 4.94072 139.871 139.871 139.871

Isonychia b. Dobbs et al.1994

0.223 110 -1.5939 0.203131 0.203131 0.203131

LC50 hardness set Z=100Y

adj LC50 SMAV GMAV

105

Keratella c. B o rgma nnand Ralph1984

0.101 180 -2.86815 0.056804 0.056804 0.056804

Lumbriculus v. West et al.1993

0.035 45 -2.57057 0.076492 0.076492 0.076492

Orconectes Dobbs et al.1994

2.37 110 0.76957 2.158837 2.158837 2.158837

Paratya Daly et al.1990a

0.034 17 -1.64643 0.192737 0.192737 0.192737

Physa integra Arthur andL e o n a r d1970

0.039 44 -2.44035 0.08713 0.08713 0.08713

Physella Dobbs et al.1994

0.109 110 -2.30973 0.099288 0.099288 0.099288

Pteronarcys N e h r i n g1976

11.8 46 3.228417 25.23966 25.23966 25.23966

Scapholeberis Carlson etal. 1986

0.018 52 -3.37711 0.034146 0.034146 0.034146

Stenonema Dobbs et al.1994

0.453 110 -0.88518 0.412638 0.412638 0.412638

Atlantic salmon Carson andCarson 1972

0.025 14 -1.76381 0.171390.205873

Salmo sp.0.145282

Atlantic salmon Sprague andR a m s a y1965

0.032 14 -1.51695 0.219379

Atlantic salmon S p r a g u e1964a

0.048 20 -1.46072 0.23207

cutthroat trout Chakoumakis et al. 1979

0.0157 26.4 -2.85009 0.0578390.102523

cutthroat trout Chakoumakis et al. 1979

0.367 205 -1.70525 0.181728

Blacknose dace Geckler etal. 1976

0.32 200 -1.81811 0.162332 0.162332 0.162332

bluegill Blaylock etal. 1985

1.3 85 0.42149 1.524231

bluegill Tho mpsonet al. 1980

1 35 1.027905 2.795204

bluegill Benoit 1975

1.1 45 0.877147 2.404032

bluegill Dobbs et al.1994

4.3 110 1.365295 3.9168772.2938

sunfish1.511977

bluegill Trama 1954 0.74 46 0.459212 1.582826

LC50 hardness set Z=100Y

adj LC50 SMAV GMAV

106

pumpkinseeds Spear andA n d e r s o n1975

1.24 125 -0.00337 0.996632 0.996632

Brook trout McKim andBenoit 1971

0.1 45 -1.52075 0.218548 0.218548 0.218548

brown bullhead Brungs et al.1973

0.182 202 -2.39217 0.0914310.15826 0.15826

brown bullhead Geckler etal. 1976

0.54 200 -1.29486 0.273936

Ch. rainbowfish Williams etal. 1991

0.179 27 -0.43837 0.645086 0.645086 0.645086

chiselmouth Andros andGarton 1980

0.143 24 -0.54759 0.578343 0.578343 0.578343

creek chub Geckler etal. 1976

0.31 200 -1.84986 0.157259 0.157259 0.157259

eel-t catfish Williams etal. 1991

0.085 27 -1.18311 0.306326 0.306326 0.306326

F a t h e a dminnow

Nelson et al.1986

0.008 49.2 -4.13384 0.016021

F a t h e a dminnow

Nelson et al.1986

0.075 47.2 -1.85516 0.156427

F a t h e a dminnow

Pickering etal. 1977

0.469 202 -1.44557 0.235611

F a t h e a dminnow

Pickering etal. 1977

0.47 202 -1.44344 0.236114

F a t h e a dminnow

Mount andS t e p h a n1969

0.075 31 -1.44353 0.236092

F a t h e a dminnow

Geckler etal. 1976

0.44 200 -1.49966 0.223207

F a t h e a dminnow

Geckler etal. 1976

0.49 200 -1.39203 0.248571

F a t h e a dminnow

Dobbs et al.1994

0.284 110 -1.3521 0.258696

F a t h e a dminnow

Nelson et al.1986

0.079 45 -1.75647 0.172653

F a t h e a dminnow

Nelson et al.1986

0.048 255 -3.9531 0.019195

F a t h e a dminnow

Nelson et al.1986

0.119 243 -2.99799 0.049887

F a t h e a dminnow

Nelson et al.1986

0.06 45.1 -2.03375 0.130844

0.12459minnow

0.132944F a t h e a dminnow

Nelson et al.1986

0.157 45 -1.06967 0.343121

F a t h e a dminnow

Nelson et al.1986

0.022 45 -3.03488 0.048081

LC50 hardness set Z=100Y

adj LC50 SMAV GMAV

107

F a t h e a dminnow

Nelson et al.1986

0.068 44 -1.88441 0.151919

F a t h e a dminnow

Nelson et al.1986

0.095 44 -1.55004 0.21224

F a t h e a dminnow

Nelson et al.1986

0.022 46.2 -3.06064 0.046858

F a t h e a dminnow

Nelson et al.1986

0.032 47 -2.70276 0.06702

F a t h e a dminnow

Carlson etal. 1986

0.055 52 -2.26015 0.104335

F a t h e a dminnow

Spehar andFiandt 1986

0.096 43.9 -1.53734 0.214952

F a t h e a dminnow

Nelson et al.1986

0.051 44 -2.17209 0.113939

F a t h e a dminnow

Nelson et al.1986

0.023 46.2 -3.01619 0.048987

F a t h e a dminnow

Nelson et al.1986

0.082 45.2 -1.72354 0.178433

F a t h e a dminnow

Nelson et al.1986

0.066 45 -1.93626 0.144242

F a t h e a dminnow

Erickson etal. 1996

0.074 45 -1.82185 0.161726

F a t h e a dminnow

Jop et al.1993

0.18 52 -1.07452 0.34146

B l u n t n o s eminnow

Horning a n dN e i h e i s e l1979

0.23 200 -2.14835 0.1166760.141859

B l u n t n o s eminnow

Geckler etal. 1976

0.34 200 -1.75749 0.172478

Flagfish Fogels andS p r a g u e1977

1.27 362 -1.0206 0.360379 0.360379 0.360379

goldfish T sa i andMcKee 1980

0.3 52 -0.5637 0.5691 0.5691 0.5691

guppies Chynowethet al. 1976

0.112 67 -1.79714 0.165772

guppies Chynowethet al. 1976

0.138 88 -1.85534 0.1564 0.146095 0.146095

guppies A n d e r s o nand Weber1975

0.223 144 -1.85761 0.156045

guppies Spear andA n d e r s o n1975

0.139 124 -2.1839 0.112601

LC50 hardness set Z=100Y

adj LC50 SMAV GMAV

108

n o r t h e r nsquawfish

Andros andGarton 1980

0.02 24 -2.5147 0.080887 0.080887 0.080887

o r a n g e t h r .darter

Geckler etal. 1976

0.85 200 -0.8412 0.431195 0.431195 darter0.264569

rainbow darter Geckler etal. 1976

0.32 200 -1.81811 0.162332 0.162332

pennyfish Williams etal. 1991

0.077 27 -1.28195 0.277495 0.277495 0.277495

chinook salmon F i n l a y s o nand Verrue1982

0.032 21 -1.91395 0.1474960.065911

Oncorhynchus0.097892

chinook salmon C h a p m a na n dM c C r a d y1977

0.01 13 -2.60754 0.073715

chinook salmon C h a p m a na n dM c C r a d y1977

0.025 46 -2.92856 0.053474

chinook salmon C h a p m a na n dM c C r a d y1977

0.09 182 -2.99428 0.050073

chinook salmon C h a p m a na n dM c C r a d y1977

0.125 359 -3.33091 0.035761

chinook salmon C h a p m a n1978

0.019 23 -2.52432 0.080113

king salmon H a z e l andMeith 1970

0.029 44 -2.73662 0.064789

coho salmon L o r z a n dMcPherson1976

0.067 95 -2.65284 0.070451 0.070451

pink salmon Servizi andM a r t e n s1978

0.087 83.1 -2.26059 0.104289 0.104289

rainbow trout Hickie et al.1993

0.018 10.3 -1.79181 0.166658

rainbow trout Hickie et al.1993

0.019 10.3 -1.73774 0.175917

rainbow trout G i l e s a n dKlaverkamp1982

0.4 44 -0.11245 0.893642

rainbow trout S p e a r a n dA n d e r s o n1975

0.21 125 -1.77913 0.168784

LC50 hardness set Z=100Y

adj LC50 SMAV GMAV

109

rainbow trout Brown andDalton 1970

0.75 240 -1.14487 0.318264

rainbow trout Fogels andS p r a g u e1977

0.102 362 -3.5424 0.028944

rainbow trout Brown et al.1974

0.58 250 -1.44189 0.236481

rainbow trout C a l a m a r ia n dM a r c h e t t i1973

0.89 300 -1.19221 0.30355

rainbow trout Miller andM a c k a y1980

0.048 49 -2.3381 0.096511

rainbow trout S p e a r a n dA n d e r s o n1975

0.2 125 -1.82792 0.160747

rainbow trout S p e a r a n dA n d e r s o n1975

0.19 125 -1.87922 0.15271

rainbow trout Dixon 1980 0.323 365 -2.3978 0.090918rainbow trout Howarth

and S p r a g u e1978

0.393 370 -2.21496 0.109157

rainbow trout Howarth and S p r a g u e1978

0.154 369 -3.14917 0.042888

rainbow trout Howarth and S p r a g u e1978

0.344 361 -2.32402 0.097879

rainbow trout Howarth and S p r a g u e1978

0.825 366 -1.46275 0.231599 0.119123

rainbow trout Howarth and S p r a g u e1978

0.167 364 -3.05477 0.047133 0.119123

rainbow trout Howarth and S p r a g u e1978

0.349 101 -1.06243 0.345616 0.119123

LC50 hardness set Z=100Y

adj LC50 SMAV GMAV

110

rainbow trout Howarthand S p r a g u e1978

0.059 101 -2.83996 0.058428

rainbow trout Howarthand S p r a g u e1978

0.085 100 -2.4651 0.085

rainbow trout Howarth and S p r a g u e1978

0.191 98 -1.6357 0.194816

rainbow trout Howarth andS p r a g u e1987

0.03 30 -2.32772 0.097518

rainbow trout Howarth and S p r a g u e1978

0.049 31.5 -1.88487 0.151849

rainbow trout Howarthand S p r a g u e1978

0.059 31 -1.68349 0.185725

rainbow trout Howarth and S p r a g u e1978

0.047 30 -1.87877 0.152778 0.119123

rainbow trout Marr et al.1995

0.018 25 -2.66003 0.069946

steelhead C h a p m a n1978

0.017 23 -2.63555 0.07168

steelhead Cusimano etal. 1986

0.0028 9.2 -3.54198 0.028956

steelhead Cusimano etal. 1986

0.0042 9.2 -3.13652 0.043434

steelhead Cusimano etal. 1986

0.0066 9.2 -2.68453 0.068253

Sockeye salmon Servizi andM a r t e n s1978

0.13 83.1 -1.85896 0.155835 0.155835

stone roller Geckler e tal. 1976

0.29 200 -1.91655 0.147114 0.147114 0.147114

stone loach S o l b e a n dCooper 1976

0.7 249 -1.24991 0.28653 0.28653 0.28653

striped shiner Geckler etal. 1976

1.225 200 -0.47574 0.621428 0.621428 0.621428

LC50 hardness set Z=100Y

adj LC50 SMAV GMAV

111

zebrafish Fogels andS p r a g u e1977

0.149 362 -3.16343 0.0422810.089267 0.089267

zebrafish W e i ns t e in1978

0.24 128 -1.66882 0.188469

112

C Computation Final Acute Value and intercept of the Alberta acute guideline.

Rank GMAV ln GMAV lnGMAV^2 P sqrtP1 0.024277 -3.71823 13.8252 0.023256 0.1524992 0.033231 -3.40427 11.58907 0.046512 0.2156663 0.034146 -3.37711 11.40487 0.069767 0.2641354 0.04466 -3.10868 9.663873 0.093023 0.3049975 0.056804 -13.6083 46.48302 0.232558 0.9372976 0.067757 0.0764928 0.080887 s^2 14.439759 0.08713 s 3.799967

10 0.089267 L -4.292511 0.097892 A -3.442812 0.099288 VAF 0.03197513 0.132944 CMC 0.01598814 0.145282 -8.6449715 0.14609516 0.14711417 0.15725918 0.1582619 0.16233220 0.19273721 0.20313122 0.21854823 0.26456924 0.27749525 0.2865326 0.30632627 0.36037928 0.40640129 0.41263830 0.569131 0.57834332 0.62142833 0.645086

Rank GMAV ln GMAV lnGMAV^2 P sqrtP

113

34 0.80452335 1.0202136 1.51197737 2.15883738 3.79797939 6.72259440 14.7544541 25.2396642 139.871

114

Appendix 6. Chronic copper toxicity data on freshwater fish.

Species Lifestage Test Type Toxicity Endpoint (mg Cu/L) Environmental Conditions Hardness Classification Reference

Primary Studies

Bluegill sunfish 12 cm, 30 g

larvae

FT 22mo survival,growth,spawn:NOEL 0.077; LOEL 0.16290 d survival:NOEL 0.021; LOEL 0.040growth NOEL 0.077

pH: 7-8; T: 13-28; Alk: 43+1.1; DO: 7+1.2

pH: 7-8; T: 23-28; Alk: 43+1.1; DO: 7+1.2

45+0.9 M, P Benoit 1975

Bluntnoseminnow

FT, CuSO4 60-d growth, 2nd gen survivalNOEL 0.072; LOEL 0.119egg prodLOEL 0.018

pH: 7.88-8.31; T: 16-27; Alk: 150-286; DO: >5.9 mg/L 172-230 M, P Horning andNeiheisel 1979

Brook trout eggs, 0-1 d FT 60-d growth NOEL 0.003c; LOEL 0.005hatchLOEL 0.013; NOEL 0.007mortalityLOEL 0.027; NOEL 0.013

pH: 6.6-7.1; T: 10+1; Alk: 27.8+3.8; DO: 10+0.7 37.5+7.3 M, P Sauter et al. 1976

Brook trout FT, CuSO4 60-d standing cropLOEL 0.044NOEL 0.022

pH: 7.3-7.9; T: 5.6+0.09; Alk: 42.4+1; DO: 11.4+1.2 45.4+0.8 M, P McKim et al. 1978

Brook trout eggs, 0-1 d FT 60-d growth NOEL 0.005; LOEL 0.008hatchLOEL 0.074; NOEL 0.049mortalityLOEL 0.049; NOEL 0.027

pH: 6.7-7.1; T: 10+1; Alk: 177.6+30.4; DO: 11+1.2 187+22 M, P Sauter et al. 1976

Brook trout 15 cm, 27 g FT 22 mo survival alevins/juvNOEL 0.010; LOEL 0.0178 mosurvival,growth, spawning,hatchability eggsNOEL 0.0174; LOEL 0.0325

pH: 7.5 (6.9-8); T: 11 (4.1-21); Alk: 41.6 (38.5-44);DO: aerated

45.4 (40-48) M, P McKim and Benoit1971

Channel catfish eggs, 2-3 d FT 60 d mortality, growthNOEL 0.012; LOEL 0.018

pH: 7.4-7.6; T: 22+1; Alk: 34.1+1.8; DO: 7.3+2.4 36+1.1 M, P Sauter et al. 1976

Channel catfish eggs, 2-3 d FT 60 d mortality, growthNOEL 0.013; LOEL 0.019

pH: 7.5-7.8; T: 22+1; Alk: 172.9+34.6; DO: 8.7+0.9 186.3+38.7 M, P Sauter et al. 1976

Species Lifestage Test Type Toxicity Endpoint (mg Cu/L) Environmental Conditions Hardness Classification Reference

115

Fathead minnow 1-4, 7 d SR (1/d),CuSO4

7-d growthNOEL 0.025; LOEL 0.05survivalNOEL 0.2-0.1; LOEL 0.4-0.2

pH: 8.3; Alk: 130 218 M, P Pickering andLazorchak 1995

Fathead minnow FT 1 yr egg prod. LOEL 0.037; NOEL 0.024; EC500.042spawningLOEL 0.061; NOEL 0.038

pH: 7.9 (7.5-8.2); T: avg 23-24; DO: 7.8+0.74 202 M, P Pickering et al. 1977

Fathead minnow 1-1.5 cm(start)

FT 11 mo. growth , mort.NOEL 0.033; LOEL 0.095spawning,egg prodNOEL 0.015; LOEL 0.033

pH: 7.9 (7.5-8.5); T: 16-25.5; Alk: 161 (150-170); DO:6.4

198 (182-216)

M, P Mount 1968

Fathead minnow 30 d FT, CuNO3 32 d growthMATC 0.0062

pH: 7.7 (7.2-7.9); T: 25+3; Alk: 42.7+1.3; DO: >70% 45.3+0.8 M, P Spehar and Fiandt1986

Fathead minnow Start 10-20mm, 6 wkold

FT 11 mo. growth,hatch NOEL 0.0106; LOEL 0.0184spawning,egg prod.,mort.NOEL 0.0106; LOEL 0.0184

pH: 6.9-7.2; T: 19-25; Alk: 30-31; DO: avg 7.2-7.9 31 M, P Mount and Stephan1969

Fathead minnow 4-wk old FT 1-yr ReprodNOEL 0.024; LOEL 0.037

pH: 7.8-7.9; T: 20-24.5; Alk: 96-99; DO: 7.7+0.72 204 M, P Geckler et al. 1976

Lake trout FT, CuSO4 60-d standing cropLOEL 0.042; NOEL 0.022

pH: 7.3-7.9; T: 5.5+0.1; Alk: 42.4+1; DO: 11.4+1.2 45.4+0.8 M, P McKim et al. 1978

Northern pike FT, CuSO4 30-d standing cropLOEL 0.104 mg/L

pH: 7.3-7.9; T: 15.6+0.08; Alk: 42.4+1; DO: 11.4+1.2 45.4+0.8 M, P McKim et al. 1978

Pink salmon eggs FT, CuSO4 hatchLOEL 0.055; NOEL 0.025

pH: 7.6 (7.55-7.8); T: 5.5-8.9; Alk: 62.5 (60.8-68.2);DO: Sat

83.1 (82.9-84.4)

M, P Servizi and Martens1978

Rainbow trout FT, CuSO4 30-d standing cropLOEL 0.032; NOEL 0.010

pH: 7.3-7.9; T: 10.8+0.12; Alk: 42.4+1; DO: 11.4+1.2 45.4+0.8 M, P McKim et al. 1978

Sockeye salmon eggs FT, CuSO4 hatchLOEL 0.078NOEL 0.037

pH: 7.6 (7.55-7.8); T: 5.5-8.9; Alk: 62.5 (60.8-68.2);DO: Sat

83.1 (82.9-84.4)

M, P Servizi and Martens1978

Steelhead trout eggs FT 78-d growthEC50 0.046

pH: 7.4-7.9; T: 12; Alk: 126; DO: 9.3-10 120 M, P Seim et al. 1984

Stone Loach 8.7-12.1 cm FT 63-d LC50 0.25 pH: 8.26+0.1; T: 11.9+0.2; DO: >96% sat 249 M, P Solbe and Cooper1976

Species Lifestage Test Type Toxicity Endpoint (mg Cu/L) Environmental Conditions Hardness Classification Reference

116

White sucker FT, CuSO4 30-d standing cropLOEL 0.034; NOEL 0.013

pH: 7.3-7.9; T: 14.9+0.59; Alk: 42.4+1; DO: 11.4+1.2 45.4+0.8 M, P McKim et al. 1978

Secondary Studies

Atlantic salmon 38 d eggs

0-27 d eggs

SR (1/2d), CuSO4

35 d hatchLOEL 0.04survivalLOEL 0.02

pH: 6.4; T: 10 14 N, S Grande 1967

Coho salmon 45-50mm FT, CuCl2 30-d survivalLOEC 0.0181; NOEC 0.0139

pH: 7.3 (7.1-7.5); T: 12.9 (12.1-13.8); Alk: 24.8 (19-53); DO: 9.6 (8.7-10.8)

31.9 (20-79.5)

M, S Stevens 1977

Fathead minnow 3.2-3.8 cm FT, Naturalstream water

9 mo. egg prod.NOEL 0.036; LOEL 0.18

pH: 7.5-8.5; T: 0-30; Alk: 56-248; DO: 5-13 88-352 M S Brungs et al. 1976

King salmon FT, CuSO4 27 d hatchNOEL 0.08survival NOEL 0.02; LOEL 0.04growth LOEL 0.02

pH: 6.8-7.2; T: 13-14; Alk: 21; DO: near sat 44 M,S Hazel and Meith 1970

Rainbow trout eggs, 0 d SR (1/2d),CuSO4

35d hatchLOEL 0.02

pH: 6.4; T: 10 14 N, S Grande 1967

Rainbow trout eggs 6 & 24 mo. hatch LOEL 0.019; NOEL 0.012survivalLOEL 0.019; NOEL 0.012

100 NR, S Goettl et al. 1976

Zebrafish 0.29-0.69 g FT, CuSO4 10-d egg prod/viabilityNOEL 0.01; LOEL 0.02

pH: 7.7; T: 27; Alk: 81; DO: 87% Sat 128 N/M erratic,S Weinstein 1978

Unsuitable studies

Common carp eggs CuSO4 hatchNOEL 0.1; LOEL 0.5

T: 25+1 U Kaur and Virk 1980

Goldfish 20,30-d LC50 0.12 T: 15.5 220 ?, U Calamari andMarchetti 1970

Walleye eggs, 2 d FT incubation 9-12 d, hatchNOEL 0.021; LOEL 0.047

pH: 6.8-7.3; T: 15+1; Alk: 34+1.9; DO: 9.9+0.7 35+1.8 M, U Sauter et al. 1976

Legend: Test Type: S=Static, SR=Static Replacement, FT=Flow-Through;

117

Toxicity Endpoint: T=expressed as total copper, diss= expressed as dissolved copper, Cu2+= expressed as cupric ionEnvironmental Conditions: T= Water Temperature oC, Alk=Alkalinity in mg/L CaCO3, DO=Dissolved Oxygen in mg/L unless otherwise

stated; Hard=Water Hardness in mg/L CaCO3;Classification: M=Measured Concentrations, M-D= Measured Concentration, but significant decrease over test duration,

N=Nominal, NR=Not Reported; C=Calculated; RP=ReplicatedP=Primary, S=Secondary, U=Unsuitable.

118

Appendix 7. Chronic copper toxicity data on freshwater invertebrates.

Species Lifestage Test Type Toxicity Endpoint (mg Cu/L) Environmental Conditions Hardness Classification

Reference

Primary Studies

Asellus meridianus 1-1.5 mm SR 12-20 d growth red. LOEL 0.1 HMSO sol; T: 20 25 M, P Brown 1976

Ceriodaphnia dubia <4hr SR, CuCl2 7 d survival/reprodNOEL 0.012; LOEL 0.032

pH: 7.5; T: 22-28; Alk: 38; DO: M 36 M, P Carlson et al. 1986

Ceriodaphnia dubia <24 hr SR, CuNO3 7-d reprod MATC 0.045 pH: 8.2 (8.0-8.5); T: 25+2; Alk: 97+9.3; DO: >70% sat 100+7.9 M, P Spehar and Fiandt1986

Ceriodaphnia dubia 2-8 hr SR 7-d reproductionNOEL 0.006; LOEL 0.01NOEL 0.005; LOEL 0.019

pH: 8.15+0.6; T: 25+1; Alk: 69.6+6.3pH: 8.31+0.03; T: 25+1; Alk: 140.1+3.9

94.1+3.9179+9.3

M, P Belanger et al.1989

Chironomus tentans 4th instar FT, CuCl2 20 d LC50 0.0775 development/emergenceNOEL 0.034; LOEL 0.084

pH: 7.4; T: 15; Alk: 19 36 M, P Nebeker et al.1984b

Clistoroniamagnifica

5th instarlarvae

FT, CuCl2 8 mo. adult emergenceNOEL 0.0083; LOEL 0.013

pH: 7.2-7.4; T: 15+2; Alk: 26+1 26+1 M, P Nebeker et al.1984a

Copepods(Acanthocyclops &Diacyclops)

CuCl2 growth EC20 0.042 pH: 8.3; T: 20; Alk: 103 180 M, P Borgmann andRalph 1984

Corbicula fluminea ad. 14-21mmad. 13-17mm

juv 7.2-9.3mm

FT, CuSO4 30-d LC50 0.019230-d LC60 0.017730-d growth NOEL 0.0084; LOEL 0.013930-d LC50 0.013430-d growth LOEL 0.0084

pH: 8.31-8.44; T: 23.9-24.7; Alk: 45.3-46.9pH: 8.18-8.36; T: 23.9-24.9; Alk: 59.6-60.8

70.3-71.777.3-78

M, P Belanger et al.1990

Daphnia magna <24 hr SR, CuCl2 14-d survivalEC50 0.038reprodLOEC 0.03; NOEC 0.01

Alk: 82 85 M, P Blaylock et al.1985

Daphnia magna SR, CuCl2 21-d LC50 0.069surv/reprNOEL 0.037; LOEL 0.11carapace grNOEL 0.013; LOEL 0.037

pH: 8.1; T: 20+0.5 225 M, P Van Leeuwen et al.1988

Species Lifestage Test Type Toxicity Endpoint (mg Cu/L) Environmental Conditions Hardness Classification

Reference

119

Daphnia pulex <24 hr SR 42 dNOEC 0.004; LOEC 0.006NOEC 0.005; LOEC 0.010

pH: 8.6; T: 20; Alk: 115pH: 8.7; T: 20; Alk: 115

57.5115

M, P Winner 1985

Dreissenapolymorpha

1.6-2 cm SR(1/d),CuCl2

48-hr filtration rateEC50 0.041NOEL 0.016

pH: 7.9; T: 15; DO: aerated, SAT 150 M, P Kraak et al. 1994

Hyalella azteca 0-1 wk SR (1/wk) 10-wk survivalLOEL 0.025; NOEL 0.017

pH: 7.9-8.6; T: 25; Alk: 90 130 M, P Borgmann et al.1993

Polypedilum nubifer eggs FT, CuSO4 35-d emergence/survivalNOEL 0.01; LOEL 0.02

pH: 7.9-8; T: 24+1 68 M, P Hatakeyama 1988

SecondaryStudies

Bosminalongirostris

neonate SR (1/d), CuSO4

13-17 d growth 4th repr. instarLOEL 0.018intr. growth rate LOEL 0.010

pH: 6.9-7.1; T: 20 N, S Koivisto andKetola 1995

Brachionuscalyciflorus

0-2 hr S 2-hr swimming LOEL 0.0065-hr ingestion NOEL 0.012; LOEL 0.025-hr growthNOEL 0.0025; LOEL 0.005

pH: 7.8; T: 25; EPA (1985) water; fed N, S Janssen et al. 1993

Campelomadecisum

FT 6 wk survival NOEL 0.008; LOEL 0.0145

M-D, S Arthur andLeonard 1970

Ceriodaphnia dubia neonate 16-22hr

SR (1/d) 7-d reprod.LOEL 0.1LOEL 0.2LOEL 0.4

pH: 6; T: 25; Alk: 74pH: 8; T: 25; Alk: 144pH: 9; T: 25; Alk: 122

98182114

3RP, M,few conc, S

Belanger andCherry 1990

Crayfish eggs

newlyhatched

newlyhatched

FT, CuSO4 13-14 d hatchNOEL 0.06; LOEL 0.12517-d survivalNOEL 0.06; LOEL 0.12530-d growth retardation at0.015,0.03,0.06 mg/L was 28, 91.4and 98.6 %

pH: 7.8-8.1; T: 20 100-125 N,S Hubschman 1967

Daphnia magna 12+12 hr SR, CuSO4 pop. growthNOEL 0.06; LOEL 0.08

pH: 8.2-9.5; T: 20; Alk: 100-119; DO: 8.7-11.4 130-160 N,S Winner and Farrell1976

Species Lifestage Test Type Toxicity Endpoint (mg Cu/L) Environmental Conditions Hardness Classification

Reference

120

Daphnia magna SR (7-d), CuCl2

3 wk LC50 0.044EC50 (reproductive impairment)0.035 mg/LNOEL 0.022

pH: 7.7; T: 18; Alk: 42; DO: near sat 45 N,S Biesinger andChristensen 1972

Daphnia magna 6-24 hr CuSO4 21-d survivalNOEL 0.0004; LOEL 0.0008reproductionLOEL 0.0004

pH: 8-8.1; T: 20.5-21; DO: sat 250 N,S Dave 1984

Daphnia similis 24-hr SR (1/d),CuSO4

28-d growthLOEL 0.001reproductionNOEL 0.01; LOEL 0.005

- N,S Soundrapandia andVenkataraman1990

Daphnia magna <24 hr SR (1/3d), CuSO4

>70 d survivalNOEL 0.04-c; LOEL 0.06-0.02reproductionNOEL 0.02-c; LOEL 0.04pop. growthNOEL 0.06-0.04; LOEL 0.08-0.06

pH: 7.2-9.5; T: 20+1; Alk: 100-118; DO: 7.3-11.4 N,S Winner et al. 1977

Daphnia ambigua 12+12 hr SR, CuSO4 pop. growthNOEL 0.04; LOEL 0.06

pH: 8.2-9.5; T: 20; Alk: 100-119; DO: 8.7-11.4 130-160 N,S Winner and Farrell1976

Daphnia pulex 12+12 hr SR, CuSO4 pop. growthNOEL 0.04; LOEL 0.06

pH: 8.2-9.5; T: 20; Alk: 100-119; DO: 8.7-11.4 130-160 N,S Winner and Farrell1976

Daphnia parvula 12+12 hr SR, CuSO4 pop. growthNOEL 0.04; LOEL 0.06

pH: 8.2-9.5; T: 20; Alk: 100-119; DO: 8.7-11.4 130-160 N,S Winner and Farrell1976

Gammaruspseudolimnaeus

FT 9 wk 2nd generation growthNOEL 0.0046; LOEL 0.0086 wk growth and survivalNOEL 0.008; LOEL 0.0148

M-D, S Arthur andLeonard 1970

Paratanytarsusparthenogeniticus

7-d S, CuSO4 18-d growthNOEL 0.32; LOEL 0.6418-d egg prod.NOEL 0.08; LOEL 0.16

pH: 6.9-7.1; T: 23+1 25 M-D, S Hatakeyama andYasuno 1981

Physa integra FT 6-wk growth and survivalNOEL 0.008; LOEL 0.0148

M-D, S Arthur andLeonard 1970

121

Legend: Test Type: S=Static, SR=Static Replacement, FT=Flow-Through;Toxicity Endpoint: T=expressed as total copper, diss= expressed as dissolved copper, Cu2+= expressed as cupric ionEnvironmental Conditions: T= Water Temperature oC, Alk=Alkalinity in mg/L CaCO3, DO=Dissolved Oxygen in mg/L unless otherwise stated;

Hard=Water Hardness in mg/L CaCO3;Classification: M=Measured Concentrations, M-D= Measured Concentration, but significant decrease over test duration, N=Nominal,

NR=Not Reported; C=Calculated; RP=ReplicatedP=Primary, S=Secondary, U=Unsuitable.

122

Appendix 8 Computations for the chronic Alberta copper guideline.

A Computations Species Mean Chronic Values (SMCV) and Genus Mean Chronic Values(GMCV). MATC values are the geometric mean of the NOEC and LOEC values in mg/Land water hardness in mg/L CaCO3.

Species Reference ToxicityEndpoint

waterhardness

NOEL LOEL MATC SMCV GMCV

bluegill sunfish Benoit 1975 survival 45 0.021 0.04 0.028983 0.028983 0.028983brook trout Sauter et al. 1976 growth 37.5 0.003 0.005 0.003873

0.009984Salvelinus0.017421brook trout McKim et al. 1978 standing crop 45.4 0.022 0.044 0.031113

brook trout Sauter et al. 1976 growth 187 0.005 0.008 0.006325brook trout M c K i m a n d

Benoit 1971survival 45.4 0.01 0.017 0.013038

lake trout McKim et al. 1978 standing crop 45.4 0.022 0.042 0.030397 0.030397channel catfish Sauter et al. 1976 growth/surv. 36 0.012 0.018 0.014697 0.015198 0.015198channel catfish Sauter et al. 1976 growth/surv. 186 0.013 0.019 0.015716fathead minnow P i c k e r i n g a n d

Lazorchak 1995growth 218 0.025 0.05 0.035355 0.019812 Minnow

0.013202fathead minnow Pickering et al.

1977egg prod. 202 0.024 0.037 0.029799

fathead minnow Mount 1968 egg prod. 198 0.015 0.033 0.022249fathead minnow Spehar and Fiandt

1986growth 45.3 0.0062

fathead minnow M o u n t a n dStephan 1969

growth etc. 31 0.0106 0.0184 0.013966

fathead minnow Geckler et al. 1976 reprod. 204 0.024 0.037 0.029799bluntnose minnow H o r n i n g a n d

Neiheisel 1979egg prod. 200 0.0043 0.018 0.008798 0.008798

northern pike McKim et al. 1978 standing crop 45.4 0.0349 0.104 0.060246 0.060246 0.060246pink salmon S e r v i z i a n d

Martens 1978growth 83.1 0.025 0.055 0.037081 0.037081 Oncorhynchus

0.024677sockeye salmon Servizi and Martens

1978growth 83.1 0.037 0.078 0.053722 0.053722

rainbow trout McKim et al. 1978 standing crop 45.4 0.01 0.032 0.017889 0.007544rainbow trout Marr et al. in press growth 24.8 0.0022 0.0046 0.003181white sucker McKim et al. 1978 standing crop 45.4 0.013 0.034 0.021024 0.021024 0.021024Ceriodaphnia Carlson et al. 1986 surv/reprod 36 0.012 0.032 0.019596 0.016063 0.016063Ceriodaphnia Spehar and Fiandt

1986reprod 100 0.045

Ceriodaphnia Belanger et al. 1989

reprod. 179 0.005 0.019 0.009747

Ceriodaphnia Belanger et al. 1989

reprod. 94.1 0.006 0.01 0.007746

Chironomus Nebeker et al.1984b

emergence 36 0.034 0.084 0.053442 0.053442 0.053442

Species Reference ToxicityEndpoint

waterhardness

NOEL LOEL MATC SMCV GMCV

123

Clistoronia Nebeker et al.1984a

emergence 26 0.0083 0.013 0.010387 0.010387 0.010387

Corbicula Belanger et al. 1990growth 77.7 0.0084 0.0139 0.010806 0.010806 0.010806Daphnia magna Blaylock et al. 1985reprod 85 0.01 0.03 0.017321 0.033241 Daphnia

0.013987Daphnia magna Van Leeuwen et al.1988

surv/reprod 225 0.037 0.11 0.063797

Daphnia pulex Winner 1985 surv. 57.5 0.004 0.006 0.004899 0.005886Daphnia pulex Winner 1985 surv. 115 0.005 0.01 0.007071Hyalella Borgmann et al.

1993surv. 130 0.017 0.025 0.020616 0.020616 0.020616

Polypedilum Hatakeyama 1988 emerg./surv 68 0.01 0.02 0.014142 0.014142 0.014142

B Computation Final Chronic Value and the Alberta chronic guideline.

Rank GMCV ln ln^2 P sqrtP1 0.010387 -4.56715 20.85889 0.066667 0.2581992 0.010806 -4.5277 20.50002 0.133333 0.3651483 0.013202 -4.32735 18.726 0.2 0.4472144 0.013987 -4.2696 18.22947 0.266667 0.5163985 0.014142 -17.6918 78.31437 0.666667 1.5869596 0.0151987 0.016063 s^2 1.7384288 0.017421 s 1.3184949 0.020616 L -4.94605

10 0.021024 A -4.6512211 0.024677 FCV 0.0095512 0.02898313 0.05344214 0.060246