United States Office of Water EPA 822

- R

- 13

- 001

Environmental Protection 4304T April 201 3

Agency



# AQUATIC LIFE AMBIENT WATER

QUALITY CRITERIA FOR

AMMONIA

– FRESHWATER

201 3

## ii EPA

- 822

- R

- 1 3

- 001



# Aquatic Life

Ambient Water Quality Criteria For

Ammonia

– Freshwater

201 3

April 2013

U.S. Environmental Protection Agency

Office of Water

Office of Science and Technology

Washington, DC

## iii 

# TABLE OF CONTENTS

TABLE OF CONTENTS

................................ ................................ ................................ ............... iii

LIST OF TABLES

................................ ................................ ................................ ........................... v

LIST OF FIGURE S

................................ ................................ ................................ ....................... vi

LIST OF APPENDICES

................................ ................................ ................................ ............... vii

FOREWORD

................................ ................................ ................................ ............................... viii

ACKNOWLEDGMENT

................................ ................................ ................................ ................ ix

EXECUTIVE SUMM ARY

................................ ................................ ................................ ............. x

ACRONYMS

................................ ................................ ................................ ............................... xiii

INTRODUCTION AND BACKGROUND

................................ ................................ .................... 1

PROBLEM FORMULATION

................................ ................................ ................................ ......... 4

Overview of Stressor Sources and Occurrence

................................ ................................ ........... 5

Environmental Fate and Transport of Ammonia in the Aquatic Environment

........................... 6

Mode of Action and Toxicity

................................ ................................ ................................ ...... 8

Assessment Endpoints

................................ ................................ ................................ ................ 9

Measures of Effect

................................ ................................ ................................ .................... 10

Acute measures of effect

................................ ................................ ................................ ....... 11

Chronic measures of effect

................................ ................................ ................................ ... 12

Chronic averaging peri od of 30 days

................................ ................................ .................... 13

Ammonia toxicity data fulfilling minimum data requirements

................................ ............ 14

Conceptual Model

................................ ................................ ................................ ..................... 16

Conceptual diagram

................................ ................................ ................................ .............. 17

Analysis P lan

................................ ................................ ................................ ............................ 19

EFFECTS ANALYSES FOR FRESHWATER AQUATIC ORGANISMS

................................ . 21

Acute Toxicity to Aquatic Animals

................................ ................................ .......................... 21

Summaries of studies used in acute criterion determination

................................ ................. 24

Chronic Toxicity to Freshwater Aquatic Animals

................................ ................................ .... 31

Summaries of studies used in chronic criterion det ermination

................................ ............. 34

The National Criteria for Ammonia in Fresh Water

................................ ................................ . 40

Acute criterion calculations

................................ ................................ ................................ .. 40

Chronic criterion calculations

................................ ................................ ............................... 46

Additional explanation and justification supporting the 2013 temperature and pH

- dependent

calcul ations and criteria magnitudes

................................ ................................ ..................... 50

Protection of downstream waters

................................ ................................ .......................... 51

Considerations for site

- specific criteria derivation

................................ ............................... 52

EFFECTS CHARACTERIZATION

................................ ................................ ............................. 52

Fres hwater Acute Toxicity Data

................................ ................................ ............................... 53

Acute toxicity data for freshwater mussels and non

- pulmonate (gill

- bearing) snails

........... 53

Freshwater Chronic Toxicity Data

................................ ................................ ............................ 56

Use of 28

- day juvenile unionid mussel data

................................ ................................ ......... 56

28

- day toxicity data for freshwater snails

................................ ................................ ............. 56

## iv

28

- day toxicity data for Hyalella azteca : Minimum Data Requirement Num ber 5

.............. 57

Reconsideration of the chronic toxicity data available for aquatic insects: Minimum Data

Requirement Number 6

................................ ................................ ................................ ......... 59

New chronic toxicity data for salmonid species and derivation of a GMCV for

Oncorhynchus: Minimum Data Requirement Number 1

................................ ...................... 59

Another order of insect or a phylum not already represented: Minimum Data Requirement

Number 8

................................ ................................ ................................ .............................. 61

Protection of Endangered Species

................................ ................................ ............................ 62

Key acute toxicity data for li sted species

................................ ................................ .............. 62

Key chronic toxicity data for listed species

................................ ................................ .......... 64

Comparison of 1999, 2009, and 2013 Criteria Values

................................ ............................. 65

Comparison of statistical approaches to develop the chronic criterion: EC20 vs. MATC

... 68

UNUSED DATA

................................ ................................ ................................ ........................... 68

REFERENCES

................................ ................................ ................................ .............................. 70

## v 

# LI ST OF TABLES

Table 1. Summary of Assessment Endpoints and Measures of Effect Used in Criteria

Derivation for Ammonia.

................................ ................................ ................................ ... 13

Table 2. 1985 Guidelines Minimum Data Requirements Summary Table Reflecting the

Number of Species and Genus Level Mean Values Represe nted in the Acute and

Chronic Toxicity Datasets for Ammonia in Freshwater.

................................ ................... 15

Table 3. Ranked Genus Mean Acute Values.

................................ ................................ ............... 27

Table 4. Ranked Genus Mean Chronic Values.

................................ ................................ ............ 39

Table 5a. Temperature and pH

- Dependent Values of the CMC (Acute Criterion Magnitude)

– Oncorhynchus spp. Present.

................................ ................................ ............................ 44

Table 5b. Temperature and pH

- Dependent Values of the CMC (Acute Criterion Magnitude)

– Oncorhynchus spp. Absent.

................................ ................................ ............................ 45

Table 6. Temperature and pH

- Dependent Values of the CCC (Chronic Criterion

Magnitude).

................................ ................................ ................................ ........................ 49

Table 7. Comparison of the Four Taxa Used to Calculate the FAV and CMC in the 1999,

2009 Draft and 2013 AWQC.

................................ ................................ ............................ 55

Table 8. Comparison of the Four Taxa Used to Calculate the FCV and CCC in the 1999

Update, 2009 Draft and the 2013 AWQC.

................................ ................................ ......... 67

Table F.1. Species, Genus and Taxon

- Specific ACRs for Freshwater Aquatic Animals

Exposed to Ammonia

................................ ................................ ................................ ....... 145

Table M.1. Results of Regression Analysis of logLC 50 

## (mg/L total ammonia nitrogen)

Versus Temperature (°C) for Individual Data Sets on the Temperature Dependence

of Acute Ammonia Toxicity.

................................ ................................ ........................... 215

Table M.2. Results of Regression Analysis of log LC 50 

## (mg/L total ammonia nitrogen)

Versus Temperature (°C) for Pooled Data Sets on the Temperature Dependence of

Acute Ammonia Toxicity to Fish.

................................ ................................ ................... 216

Table N.1. Temperature and pH

- Dependent Values of the CMC (Acute Criterion

Magnitude)

– Unionid Mussels Present, Oncorhynchus Absent.

................................ .... 233

Table N.2. Acute Data Without Mussels: Comparison of the Four Taxa Used to Calculate

the FAV and CMC in the 1999 AWQC and this Updated 2013 AWQC Excluding

Data for Freshwater Unionid Mussels.

................................ ................................ ............ 234

Table N.3. Temperature and pH

- Dependent Values of the CMC (Acute Criterion

Magnitude)

– Unionid Mussels Absent and Oncorhynchus Present.

.............................. 23 5

Table N.4. Temperature and pH

- Dependent Values of the CMC (Acute Criterion

Magnitude)

– Unionid Mussels Absent and Oncorhynch us Absent.

............................... 236

Table N.5. 2013 Acute Criterion Recalculations for Site

- specific Criteria.

............................... 237

Table N.6. Chronic Dataset Without Mussels: Comparison of the Four Taxa used to

Calculate the CCC in the 1999 AWQC and this Updated 2013 AWQC Excluding

Data for Freshwater Unionid Mussels.

................................ ................................ ............ 237

Table N.7. Chronic Criterion Recalculations for Site

- Specific Criteria.

................................ .... 238

Table N.8. Temperature and pH

- Dependent Values of the CCC (Chronic Criterion

Magnitude)

– Mussels Absent and Early Life Stage (ELS) Protection Necessary.

......... 239

Table N.9. Temperature and pH

- Dependent Values of the CCC (Chronic Criterion

Magnitude)

– Mussels Absent and Early Life Stage ( ELS) Protection not Necessary.

... 240

## vi 

# LIST OF FIGURES

Figure 1. Fraction of Chemical Species of Ammonia Present with Change in pH (at 25°C).

........ 7

Figure 2: Conceptual Model for Ammonia Effects on Aquatic Animals.

................................ .... 18

Figure 3. Ranked Freshwater Genus Mean Acute Values (GMAVs) with Criterion

Maximum Concentrations (CMCs).

................................ ................................ ................... 24

Figure 4. Ranked Freshwater Genus Mean Chronic Values (GMCVs) with Criterion

Continuous Concentrations (CCCs).

................................ ................................ .................. 33

Figure 5a. 2013 Acute Criterion Magnitudes Extrapolated Across a Temperature Gradient

at pH 7.

................................ ................................ ................................ ............................... 43

Figure 5b. 2013 Chronic Criterion Magnitudes Extrapolated Across a Temperature Gradient

at pH 7.

................................ ................................ ................................ ............................... 48

Figure F.1. SMACRs by SMAV Rank.

................................ ................................ ...................... 148

Figure M.1. The Effect of Temperature on Ammonia Toxicity in Terms of Unionized

Ammonia (DeGraeve et al. 1987).

................................ ................................ ................... 203

Figure M.2. The Effect of Temperature on Acute Ammonia Toxicity in Terms of Total

Ammonia Nitrogen.

................................ ................................ ................................ ......... 205

Figure M.3. The Effect of Temperature on pH

- Adjusted Acute Ammonia Toxicity in Terms

of Total Ammonia Nitrogen.

................................ ................................ ............................ 206

Figure M.4. The Effect of Temperature on Normalized Acute Ammonia Toxicity in Terms

of Total Ammonia Nitrogen.

................................ ................................ ............................ 210

Figure M.5. The Effect of Temperature on Chronic Ammonia Lethality to Fathead

Minnows in Terms of Total Ammonia Nitrogen (DeGraeve et al. 1987).

...................... 211

Figure M.6. Temperature Dependence of Acute Ammonia Toxicity to Invertebrate

Organisms from Arthur et al. (1987).

................................ ................................ .............. 221

Figure M.7. Temperature

- Dependence of Ammonia ACRs for Fathead Minnows.

.................. 223

Figure N.1. Comparison of the 2013 CMC Extrapolated Across a Temperature Gradient at

pH 7 Accounting for the Presence or Absence of Unionid Mussels and the Presence

or Absence of Oncorhynchus

. ................................ ................................ .......................... 241

Figure N.2. Comparison of the 2013 CCC Extrapolated Across a Temperature Gradient at

pH 7 Accounting for the Presence or Abse nce of Mussels and/or the Need for Early

Life Stage (ELS) Protection of Fish Species.

................................ ................................ .. 242

## vii 

# LIST OF APPENDICES

Appendix A. Acute Toxicity of Ammonia to Aquatic Animals.

................................ .................. 86

Appendix B. Chronic Toxicity of Ammonia to Aquatic Animals.

................................ ............. 131

Appendix C. Other Chronic Ammonia Toxicity Data.

................................ ............................... 134

Appendix D. Conversion of Acute Results of Toxicity Tests.

................................ ................... 138

Appendix E. Conversion of Chronic Results o f Toxicity Tests.

................................ ................. 141

Appendix F. Acute

- Chronic Ratios (ACRs).

................................ ................................ .............. 143

Appendix G. Results of the Regression Analyses of New Chronic Data for Unionid

Mussels.

................................ ................................ ................................ ........................... 149

Appendix H. Detailed Descriptions of Select New Acute and Chronic Toxicity Test Data

Used for Criteria Derivation.

................................ ................................ ........................... 154

Appendix I. Qualitative Weight

- of

- Evidence Test Data.

................................ ............................ 163

Appendix J. Unused Acute Studies Potentially Influential for Freshwater Ammonia Criteria

Development.

................................ ................................ ................................ ................... 171

Appendix K. Unused Chronic Studies Potentially Influential for Freshwater Ammonia

Criteria Development.

................................ ................................ ................................ ...... 177

Appendix L. Unused (Non

- Influential) Acute and Chronic Studies for Freshwater Ammonia

Criteria Development

– Screened Out Studies with Code List.

................................ ....... 179

Appendix M. 1999 Re

- examination o f Temperature Dependence of Ammonia Toxicity.

........ 199

Appendix N. Site

- Specific Criteria for Ammonia.

................................ ................................ ..... 225

## viii



# FOREWORD

This water quality criteria update provides scientific recommendations to states and tribes

authorized to establish water quality standards under the Clean Water Act (CWA), to protect

aquatic life from acute and chronic effects of ammonia in freshwater ecosystems . Under the

CWA, states and tribes are to establish water quality criteria to protect designated uses. State

and tribal decision makers retain the discretion to adopt approaches on a case

- by

- case basis that

differ from those used in th ese criteria when appropriate. While this update constitutes United

States Environmental Protection Agency (EPA) scientific recommendations regarding ambient

concentrations of ammonia that protect freshwater aquatic life, this update does not substitute for

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

binding requirements on EPA, states, tribes, or the regulated community, and might not apply to

a particular situation based upon the circumstances. EPA may change th ese crite r ia in the future

,

as new scientific information becomes available

. This document has been approved for

publication by the O ffice of Science and Technology, Office of Water, U.S. Environmental

Protection Agency. Mention of trade names or commercial products does not constitute

endorsement or recommendation for use.

## ix



# ACKNOWLEDGMENT

This 201 3 criteria document is an update of t he 1999 Update of Ambient Water Quality

Criteria for Ammonia. The updates described herein were prepared by Lisa Huff (EPA Team

Leader ), Charles Delos, Kathryn Gallagher, and Joe Beaman with written and technical support

provided b y EPA contractor: Great Lakes Environmental Center, Inc. EPA received substantial

input from Dave Mount, James (Russ) Hockett , Russell Erickson, and Charles Stephan of the

EPA’s Office of Research and Development (ORD) National Health and Environmental Effects

Research Laboratory ( NHEERL ) Mid

- Continent Ecology Division, Duluth, MN, and Cindy

Roberts, ORD Office of Science Policy . Please submit comments or questions to: Lisa Huff,

U.S. EPA, Mail Code 4304, Washington, DC 20460 (e

- mail: huff.lisa@epa.gov).

## x



# EXECUTIVE SUMM ARY

EPA has updat ed the freshwater a mm onia aquatic life ambient water quality criteria in

accord with the provisions of Section 304(a) of the Clean Water Act to revise Ambient Water

Quality Criteria (A W QC ) from time to time in order to reflect the latest s cientific knowledge

.

L iterature searches for laboratory toxicity tests of ammonia on freshwater aquatic life

, published

from 1985 to 2012

, identified new studies contain in g a cute and chronic toxicity data acceptable

for criteria derivation

. The acute criterion dataset includes 1 2 species of aquatic animals

Federally

- listed as threatened, endangered or species of concern. In the chronic dataset for

ammonia, Federally

- listed species are represented by three salmonid fish species in the genus

Oncorhynchu s , including sockeye salmon , rainbow trout/steelhead, and the subspecies Lahontan

cutthroat trout. Data were assessed from the perspective of EPA’s “ Guidelines for Deriving

Numerical National Water Quality Criteria for the Protection of Aquatic Organisms and Their

Uses ” (Stephan et al. 1985).

The 1999 recommended aquatic life criteria for ammonia were based on the most

sensitive endpoints known at the time: the acute criterion was based primarily on effects on

salmonids (where present) or other fish , and t he chronic criterion was based primarily on

reproductive effects on the benthic invertebrate Hyale l la or on survival and growth of fish early

life stages (when present), depending on temperature and season

.

The 2013 recommended criteria of this document ta ke into account data for several

sensitive f

reshwater m ussel species in the Family Unionidae that had not previously been tested

.

As noted in the 2009 draft ammonia criteria document, available data indicate d that another

freshwater mollusk tax on

, non

- pulmonate (gill

- bearing) snails, are also sensitive to the effects of

ammonia (EPA

- 822

- D

- 09

- 001)

. The 2013 criteria include additional data confirming the

sensitivity of freshwater non

- pulmonate snails. M any states in the continental United States ha ve

freshwater unionid mussel fauna in at least some of their waters (Abell et al. 2000, Williams et

al. 1993, Williams and Neves 1995). Moreover, approximately one

- quarter of approximately

300 freshwater unionid mussel taxa in the United States are Federa lly

- listed as endangered or

threatened species

. F reshwater mussels are broadly distributed across the U.S., as are freshwater

non

- pulmonate snails, another sensitive invertebrate taxon, and both of these groups are now

included in the ammonia dataset . Th us, th e 201 3 freshwater acute and chronic aquatic life

criteria for ammonia will more full y protect the aquatic community than previous criteria , and

## xi

are represented by a single (non

- bifurcated) value each for acute and chronic criteria

.

The criteria magnitude is affected by pH and temperature. After analysis of the new data,

EPA determined that the pH and temperature relationships established in the 1999 ammonia

criteri on document still hold. W hen expressed as total ammonia nitrogen (TAN), the effect

concentrations for fish are normalized only for pH

, reflecting the minimal influence of

temperature on TAN toxicity to fish. For invertebrates, TAN effect concentrations are

normalized for both pH and temperature

. A t water temperatures greater than 15.7 ° C

, the 2013

acute criteri on magnitude is determined primarily by effects on freshwater unionid mussels. At

lower temperatures the acute criterion magnitude i s based primarily on effects on salmonid s and

other fish

. T hroughout the temperature range, t he 2013 chronic criterion magnitude is

determined primarily by the effects on freshwater mollusks, particularly unionid mussels.

At an example pH of 7 and temperature of 20 ° C

, the 2013 acute criterion magnitude is 1 7

mg TA N/L and the chronic criterion magnitude is 1. 9 mg TA N/L

. A t pH 7 and 20 ° C the 201 3

acute criteri on magnitude is 1. 4

- fold lower than the 1999 acute criteri on magnitude

. At this pH

and temperature, the 201 3 chronic criterion magnitude is 2. 4

- fold lower than the 1999 chronic

criteri on magnitude

. See the Criterion Statement s (pages 40

- 49 ) for the criterion concentrations

at other pH and temperature conditions. The decrease s in acute and chronic criteria magnitudes

below those of 1999 re flect the inclusion of the new data discussed above

.

T he acute criterion duration represents a one

- hour average

. The chronic criterion

duration represents a 30

- day rolling averag e with the additional restriction that the highest 4

- day

average within the 3 0 days be no greater than 2.5 times the chronic criterion magnitude

. These

values are not to be exceeded more than once in 3 years on average

.

## xii

Criterion

Duration 1999 A WQ C

Update Criteria

Magnitude 2009 Draft A WQ C

Update Criteria c



## Magnitude 2013 AWQC

Update Criteria

Magnitude

pH 8.0,

(mg TAN/L) 

## pH 7.0,

T=20 ° C

(mg TAN/L) 

## pH 8.0,

T=25 ° C

(mg TAN/L) 

## pH 7.0,

T=20 ° C

(mg TAN/L) 

## pH 7.0,

T=20 ° C

(mg TAN/L)



## Acute

(1

- hr average) 5.6 a 

## 24 a 

## 2.9 19 17 a



## Chronic

(30

- d rolling

average) 1.2 4.5 b 

## 0.26 0.91 1.9 *

*Not to exceed 2.5 times CCC or 4.8 mg TAN/L (at pH 7, 20 ° C) as a 4

- day average within the 30

- days

, more than once in three years on average

.

Criteria frequency: Not to be exceeded more than once in three years on average.

a 

### Salmonids present

b 

### Based on renormalization of data to pH 7 and 20 ° C

c 

### Mussels present

## xiii



# ACRONYMS

ACR Acute

- Chronic Ratio

ASTM American Society of Testing and Materials

AWQC Ambient Water Quality Criteria

CCC Criterion Continuous Concentration

CMC Criterion Maximum Concentration

CV Chronic Value (expressed in this document as an EC20 or MATC)

CWA Clean Water Act

ECx Effect Concentration at X Percent Effect Level

LCx Lethal Concentration at X Percent Survival Level

EPA Environmental Protection Agency

ESA Endangered Species Act

FACR Final Acute

- Chronic Ratio

FAV Final Acute Value

FCV Final Chronic Value

GMACR Genus Mean Acute

- Chronic Ratio

GMAV Genus Mean Acute Value

GMCV Genus Mean Chronic Value

LOEC Lowest Observed Effect Concentration

MATC Maximum Acceptable Toxicant Concentration (expressed mathematically as the

geometric mean of the NOEC and LOEC)

NOEC No Observed Effect Concentration

SD Sensitivity Distribution

SMACR Species Mean Acute

- Chronic Ratio

SMAV Spec ies Mean Acute Value

SMCV Species Mean Chronic Value

TAN Total Ammonia Nitrogen

TRAP EPA’s Statistical Program: Toxicity Relationship Analysis Program (Version

1. 21 )

WER Water Effect Ratio

## 1



# INTRODUCTION AND BACKGROUND

National Ambient Water Quality Criteria (AWQC) are established by the United States

Environmental Protection Agency (EPA) under the Clean Water Act (CWA). EPA will review

and from time to time revise 304(a) A W QC as necessary to ensure the criteria are con sistent with

the latest scien tific information

. S ection 304(a) aquatic life criteria serve as recommendations to

states and tribes in defining ambient water concentrations that will protect against adverse

ecological effects to aquatic life resulting from exposure to a pollutant found in water from direct

contact or ingestion of contaminated water and/or food . Aquatic life criteria address the CWA

goals of providing for the protection and propagation of fish and shellfish

. When adopted into

state standar ds, these criteria can become a basis for establishing permit limits and Total

Maximum Daily Loads (TMDLs)

.

EPA first published aquatic life criteria recommendations for ammonia in 1976, followed

nine years later by the 1985 criteria revision , which used updated procedures and additional

information. The 1985 acute ammonia criterion was calculated from acute values expressed as

unionized ammonia and normalized for pH (8.0) for all freshwater aquatic animals, and

temperature (20 ° C) for freshwater fish only. Because the fraction of total ammonia that is

unionized varies with pH and temperature, the 1985 toxicity data normalizations with unionized

ammonia were necessarily structured differently than the current document’s normalizations with

total ammoni a nitrogen

. B ecause t he 1985 chronic toxicity dataset was more limited than is

available now , the 1985 chronic criterion was calculated by dividing the Final Acute Value by an

acute

- chronic ratio (ACR)

. T he 1985 acute and chronic criteria concentrations were 19 and 1.2

mg/L expressed as total ammonia nitrogen at pH 7 and temperature 20 ° 

## C for salmonids or other

coldwater species present (e.g.

, rainbow trout)

. The duration s for these criteria were one

- hour

(acute) and four

- day (chronic) averaging periods. T he 1985 freshwater acute criterion dataset

was composed of acute values from tests involving 41 species (29 fish and 12 invertebrate)

representing 34 genera (18 fish and 16 invertebrate). T he data available for invertebrates at the

time indicated they w ere not among the more acutely

- sensitive organisms to ammonia.

In 1999 EPA revised the 1985 freshwater ammonia criteria to incorporate newer data,

better models, and improved statistical methods. For its acute criterion

, t he revision included a

re

- examin ation of the temperature and pH relationships underlying the 1985 acute criterion

,

reworked from the perspective of total ammonia nitrogen rather than unionized ammonia. For its

## 2

chronic criterion

, EPA developed relationships for formulating a seasonal, pH

- and temperature

dependent relationship , in part because the chronic criteri on was based on endpoints that might

not be of concern during cold

- season conditions (e.g., fish early life stages). EPA analyzed all of

the freshwater chronic data used in the 1985 criteria document as well as newer chronic data and

was able to directly calculate a chronic criterion instead of calculating it from the acute criterion

with an ACR. EPA did not conduct a comprehensive lite rature search for and critical review of

all of the acute toxicity data published after 1985

, but focused on the chronic criteria, in response

to scientific issues raised by the public. Thus, the 1999 acute criterion relied on acute tests

reported in Tabl e 1 in the 1985 criteria document, supplemented by a limited number of newer

studies relevant to the revised pH relationship.

The 1999 criteria were based on the most sensitive endpoints known at the time: the acute

criterion was based primarily on effects on fish throughout the temperature range , and the

chronic criterion was based primarily on effects on benthic macroinvertebrates or fish early life

stages (when present), depending on temperature and season . For the 1999 acute criterion the

effect concen trations for fish were normalized for pH only, reflecting the minimal influence of

temperature on total ammonia toxicity to fish. The 1999 acute criterion was not adjusted for

temperature because invertebrates that were included in the dataset , mollusks i ncluded, were not

among the species highly sensitive to ammonia, thus, the invertebrate temperature slope did not

affect the formulation of the 1999 acute criterion. The 1999 chronic criterion was adjusted for

pH for fish and for pH and temperature for in vertebrates. The chronic averaging period was

increased from a 4

- to a 30

- day average in the 1999 update ; the rationale for this change was

based on analysis of chronic data from fathead minnow laboratory tests of different exposure

durations and exposure concentrations with “limited variability” (see detailed discussion in the

Problem Formulation of this document under Chronic Measures of Effect ) . For chronic toxicity,

the 1999 updated dataset consisted of nine values representing four invertebrate and f ive fish

genera. Two of the four most chronically sensitive species were invertebrates (the benthic

amphipod Hyalella azteca and the bivalve mollusk, Musculium transversum ). Missing were

representative chronic values for the genus Oncorhynchus (salmonid) and an insect genus,

although in both of these cases the calculation of the fifth percentile directly from the GMCVs in

Table 5 of the 1999 update was deemed to adequately protect the freshwater aquatic community.

## 3

In 2004 EPA published a Feder al Register Notice indicating its intent to re

- evaluate the

freshwater a mm onia criteria and request ing new in f or m ation on ammonia toxicity to freshwater

mussel species in the Family Unionidae

. This action was taken in response to concerns from

U.S. Fish a nd Wildlife Service ( USFWS ) and mussel researchers about the sensitivity of unionid

mussels to ammonia (summarized by Augspurger et al. 2003)

. The current document takes into

account all such data

, new toxicity data obtained by a search of the literature for all other species,

and updated analyses of tests previously included in the 1999 document.

In 2009, EPA published a draft ammonia criteria document that included all available

new data on the toxicity of ammonia to freshwater mussels (EPA

- 822

- D

- 09

- 001). The draft

2009 document incorporate d new toxicity data in the acute and chronic dataset while retaining

the relationships describing the influence of pH and temperature on ammonia toxicity es tablished

in the 1999 criteria. The 2009 ac ute dat a set represented 67 genera

, including 12 species of

freshwater mussels

, compared to o nly 34 genera in the 1999 AWQC. Freshwater bivalve

mollusk s and snails were the predominant group s of genera ranked in the lowest (most sensitive)

quartile, and the four most acutely sensitive genera were all bivalves. The 2009 chronic dataset

incorporated two new fish species and new data for three freshwater mussel species, which

represented t w o of the four most sensitive genera. The draft 2009 criteria recomm endation s

w ere bifurcated

, with a set of acute and chronic criteria values for waters with mussels present

that reflect s the ir greater sensitivity to ammonia, and a different set of criteria values for w aters

where mussels are absent. Including the new acceptable data for freshwater unionid mussels, the

draft 2009 acute and chronic criteri a magnitudes , respectively, w ere 1 9 and 0. 91 mg TAN/L

adjusted to pH 7 .0 and 2 0 ° C.

For this 2013 update, EPA conducted a new literature search for both acute and chronic

toxicity data and reanalyzed data consider ed in the 1999 criteria and the 2009 draft . EPA

reviewed results from this literature search and reanalysis of previously considered data to

identify data from laboratory toxicity tests that quantify the a dverse effects of ammonia on

freshwater aquatic life (amphibians, fishes, and macroinvertebrates ), with particular attention

given to tests conducted with freshwater unionid mussels and non

- pulmonate snails

, since such

data were not available for many of t hese species previously

. While unionid mussel species are

not prevalent in some waters

, such as in the arid west, non

- pulmonate snails are broadly

distributed across the U.S

. Thus, considering that freshwater unionid mussels are among the

## 4

most sensitive genera in the dataset, and that all states have at least one freshwater unionid

mussel or bivalve mollusk, or non

- pulmonate snail species, another relatively sensitive mollusk

group, native or present in at least some of their waters, EPA is recommending a single national

acute and a single national chronic criterion be applied to all waters rather than different criteria

based on the presence or absence of mussels

.

EPA also conducted a separate search and analysis of any relevant new data sp ecific for

freshwater mussels to evaluate whether the existing pH

- acute TAN toxicity relationship

established in the 1999 update document similarly applies to this group of invertebrates. Based

on the results of the literature review

, EPA concludes that t he same pH and temperature

relationships used to account for the influence of these two abiotic factors on ammonia toxicity

in the 1999 AWQC document are still applicable (e.g., see Additional Explanation and

Justification Supporting the 2013 Temperature a nd pH

- Dependent Calculations and Criteria

Magnitudes section for additional details , pg. 50 )

.



# PROBLEM FORMULATION

Problem formulation provides a strategic frame work for water quality criteria

development by focusing the effects assessment on the most rele vant chemical properties and

endpoints. The structure of this effects assessment is consistent with EPA’s Guid elines for

Ecological Risk Assessment (U.S. EPA 1998)

This ecological effects assessment defines scientifically

- defensible water quality criteri a

values for ammonia under section 304(a)(1) of the Clean Water Act. The goal of the Clean

Water A ct is to protect and restore the biological, chemical and physical integrity of waters of

the U.S. Clean Water Act Section 304(a)(1) requires EPA to develop criteria for water quality

that accurately reflect the latest scientific knowledge. These criteria are based solely on data and

best professional scientific judgments on toxicological effects. Criteria are developed following

the guidance outlined in th e Agency’s Guidelines for Deriving Numerical National Water

Quality Criteria for the Protection of Aquatic Organisms and Their Uses (Stephan et al. 1985).

Once S ection 304(a) water quality criteria are finalize d, states and authorized tribes may

adopt the criteria into their water quality standards to protect designated uses of water bodies.

States and tribes may also modify the criteria to reflect site

- specific conditions or use other

## 5

scientifically

- defensible methods to develop standards. Water quality standards are subsequently

approved by EPA.



# Overview of Stressor Sources and Occurrence

Ammonia is considered one of the most important pollutants in the aquatic environment

not only because of its highly toxic nature, but also its ubiquity in surface water systems (Russo

1985)

. Ammonia is produced for commercial fertilizers and other industrial applications using a

reaction that converts atmospheric nitrogen to ammonia using hydrogen obtained from methane

( natural gas ) under high heat and p ressure; the ammonia gas is then compressed under low

temperature and stored in an anhydrous liquid form (Appl 1999). In agriculture, ammonia is

used both directly (in anhydrous form), as well as a precursor for other nitrogen

- based fertilizers

such as am monium nitrate, ammonium pho sphate, urea, and ammonium sulf ate ( Environment

Canada 20 1 0 ) . The agricultural industry uses approximately 90% of the U.S. annual domestic

ammonia production (USGS 2004). Ammonia also has numerous industrial applications,

incl uding use as a protective atmosphere and as a source of hydrogen in metal finishing and

treating applications (e.g., nitriding; Appl 1999), as well as many other uses in the chemical

industry including the production of pharmaceuticals (Karolyi 1968) and d yes (Appl 1999). The

petroleum industry utilizes ammonia for processing of crude oil and in corrosion protection ( U.S.

EPA 2004). Ammonia is also used in the mining industry for metals extraction ( U.S. EPA

2004). Natural sources of ammonia include the d ecomposition or breakdown of organic waste

matter, gas exchange with the atmosphere, forest fires, animal waste, the discharge of ammonia

by biota, and nitrogen fixation processes (Environment Canada 1997; Environment Canada

2010; Geadah 1985).

Ammonia can enter the aquatic environment via anthropogenic sources or discharges

such as municipal effluent discharges , agricultural runoff , and natural sources such as nitrogen

fixation and the excretion of nitrogenous wastes from animals. While much of the early

information regarding lethal concentrations of ammonia was driven by the consequences of

ammonia buildup in aquaculture systems (i.e., fish culture po nds, hatchery raceways, and fish

holding and transporting tanks), the introduction of ammonia into surface water systems from

industrial processes, agricultural runoff, and sewage effluents has received considerable attention

since the 1980s (Alabaster and Lloyd 1980 ; U.S. EPA 1985). Many effluents have to be treated

## 6

extensively in order to keep the concentrations of ammonia in surface waters from being

unacceptably high. In 2011, there w ere approximately 4

. 7 million pounds ( lbs. ) of ammonia

documented as discharged from all reporting industries to surface waters ( U.S. EPA 2011). In

2010, industrial releases of ammonia to ten l arge a quatic e cosystem s (e.g., Chesapeake Bay,

Puget S ound, Great Lakes ) were r eported to total approximately 1.3 million lbs. ( U.S. EPA

2010).



# Environmental Fate and Transport of Ammonia in the Aquatic Environment

Ammonia (NH 3 

## ) is formed in the natural environment by the fixation of atmospheric

nitrogen and hydrogen by diazotrophic microbes, such as cyanobacteria (Latysheva et al. 2012)

.

T race amounts are also produced by lightning (Noxon 1976 ) . Decomposition of manure, dead

plants and animals by bacteria in the aquatic and terrestrial environments produce ammonia and

other ammonium compounds through conversion of nitrogen dur ing decomposition of tissues in

a process called ammonification (ATSDR 2004; Sylvia 2005). In the aquatic environment,

ammonia is also produced and excreted by fish

. The chemical form of ammonia in water

consists of two species, the more abundant of whic h is the ammonium ion (NH 4 + 

## ) and the less

abundant of which is the non

- dissociated or unionized ammonia (NH 3 

## ) molecule; the ratio of

these species in a given aqueous solution is dependent upon both pH and temperature (Emerson

et al. 1975 ; Erickson 1985 ; Th urston 1988 ; Whitfield 1974; Wood 1993). Chemically, ammonia

in an aqueous medium behaves as a moderately strong base with p K

a 

## values ranging from

approximately 9 to slightly above 10 as a function of temperature and ionic strength (Emerson et

al. 1975 ; Whitfield 1974). In general, the ratio of unionized ammonia to ammonium ion in fresh

water increases by 10

- fold for each rise of a single pH unit, and by approximately two

- fold for

each 10 ° C rise in temperature from 0

- 30 ° C (Erickson 1985). B asically, as values of pH and

temperature tend to increase, the concentration of NH 3 

## increases and the concentration of NH 4 +



## decreases

.

The ionized ammoni um ion ( NH 4 + 

## ) and unionized ammonia molecule ( NH 3 

## ) are

interrelated through the chemical equilibrium NH 4 +



## - OH

- 

## ↔ NH 3 

## ·H 2 

## O ↔ NH 3 

## +H 2 

## O (Emerson et

al. 1975; Russo 1985). The concentration of total ammonia (often expressed on the basis of

nitrogen as total ammonia nitrogen or TAN) is the sum of NH 4 + 

## and NH 3 

## concentrations

. I t is

total ammonia that is analytically measure d in water samples. To estimate the relative

## 7

concentrations of NH 4 + 

## and NH 3 

## from total ammonia, Emerson et al.'s (1975) formulas are

recommended ( Adams and Bealing 1994 ; Alabaster and Lloyd 198 0 ; Richardson 1997 ; Russo

1985 ). Figure 1 ( below ) shows the chemical speciation of ammonia over a range of pH levels in

ambient waters at 25 ° C

. It depicts t he 10

- fold increase in the ratio of unionized ammonia to

ammonium ion in fresh water for each rise of a single pH unit as described above. This incr ease

in unionized ammonia with increased pH is one hypothesis explaining why toxicity of total

ammonia increases as pH increases.

Figure 1 . Fraction of C hemical S pecies of A mmonia P resent with C hange in pH (at 25 ° C )

.

Each separate fraction of total ammonia can be calculated in freshwater from the

Henderson

- Hasselbach equation if the pH and p Ka are known:

NH 4 + 

## = Total ammonia/(1+ antilog (pH

- p Ka )) = Total ammonia

– NH 3 

## (Wood 1993)

and,

p Ka = 0.09018 + (2729.92/(273.2 + T)) (Emerson et al. 19 75)

where T is temperature in ° C.

## 8



# Mode of Action and Toxicity

Ammonia is unique among regulated pollutants because it is an endogenously produced

toxicant that organisms have developed various strategies to excrete, which is in large part by

passive diffusion of unionized ammonia from internal organs, such as the gills in fish. High

external unionized ammonia concentrations reduce or reverse diffusive gradients and cause the

buildup of ammonia in internal tissues and blood. U nionized ammonia may cau se toxicity to

Nitrosomonas spp. and Nitrobacter spp

. bacteria, inhibiting the nitrification process (Russo

1985). Bacterial inhibition can result in the increased accumulation of ammonia in the aquatic

environment, thereby intensifying the toxicity to be neficial bacteria and aquatic animals (Russo

1985).

The toxic action of unionized ammonia on aquatic animals, particularly in sensitive fish,

may be due to one or more of the following causes: (1) proliferation in gill tissues, increased

ventilation rates and damage to the gill epithelium (Lang et al

. 1987); (2) reduction in blood

oxygen

- carrying capacity due to progressive acidosis (Russo 1985) ; (3) uncoupling oxidative

phosphorylation causing inhibition of production and depletion of adenosine triphosphate (ATP)

in the brain (Camargo and Alonso 2006) ; (4) and the disruption of osmoregulatory and

circulatory activity disrupting normal metabolic functioning of the liver and kidneys (Arillo et.

al.1981 ; Tomasso et al. 1980).

Among invertebrates, st udies testing ammonia toxicity to bivalves, and particularly

studies with fr eshwater mussels in the family U nionidae

, have demonstrated their sensitivity to

ammonia (Augs pu rger et al. 200 3 ; Wang et al. 2007 a

, b ; Wang et al. 2008). T oxic effects of

unioniz ed ammonia to both freshwater and marine bivalves include reduc ed opening of valves

for respiration and feeding (Epifan i o and Srna 1975) ; impaired secretion of the byssus , or

anchoring thread s in bivalves (Reddy and Menon 1979) ; reduced ciliary action in b ivalves (U

. S

.

EPA 198 5 ) ; deplet ion of lipid and carbohydrate stores leading to metabolic alteration (Chetty

and Indira 1995) as well as mortality (Goudreau et al. 1993). These negative physiological

effects may lead to reductions in feeding, fecundity

, and survivorship, resulting in decreas ed

bivalve populations (Alonso and Camargo 2004 ; Constable et al. 2003 ).

## 9



# Assessment Endpoints

Assessment endpoints are defined as “explicit expressions of the actual environmenta l value

that is to be protected” and are defined by an ecological entity (species, community, or other

entity) and its attribute or characteristics ( U.S. EPA 1998). Assessment endpoints may be

identified at any level of organization (e.g.

, individual, population, community). In the context

of the Clean Water Act

, aquatic life criteria for toxics are typically determined based on the

results of toxicity tests with aquatic organisms in which unacceptable effects on growth,

reproduction, or survival occurred . This information is aggregated into a species sensitivity

analys i s that evaluates the impact on the aquatic community. Criteria are d esigned to be

protective of the vast majority of aquatic animal species in an aquatic community (i.e.,

approximately 9 5 th 

## percentile of tested aquatic an imals representing the aquatic community ) . As

a result, health of the aquatic ecosystem may be considered as an assessment endpoint i ndicated

by survival, growth, and reproduction. To assess potential effects on the aquatic ecosystem by a

particular stre ssor, and develop 304(a) aquatic life criteria under the CWA, EPA typically

requires the following:

 Acute toxicity test data (mortality, immobility, loss of equilibrium) for aquatic animals

from a minimum of eight diverse taxonomic groups. The diversity of tested species is

intended to ensure protection of various components of an aquatic ecosystem. The acute

freshwater toxicity testing requirement is fulfilled with the following eight minimum data

requirements :

 the family Salmonidae in the class Osteic hthyes

 a second family in the class Osteichthyes, preferably a commercially or

recreationally important warmwater species (e.g., b luegill, channel catfish,

etc.)

 a third family in the phylum Chordata (may be in the class Osteichthye s or

may be an amphibia n, etc.)

 a planktonic crustacean (e .g., cladoceran, copepod, etc.)

 a benthic crustacean (e.g., ostracod, is opod, amphipod, crayfish, etc.)

 an insect (e.g., mayfly, dragonfly, damselfly, stonefly, ca ddisfly,

mosquito, midge, etc.)

## 10  a family in a phylum other than Arthropoda or Chordata (e.g., Rotifera,

Annelida, Mollusca, etc.)

 a family in any order of insect or any phylum not already represented

 Chronic toxicity test data (longer

- term survival, growth, or reproduction) are required for

a minimum of three taxa in order to use acute to chronic ratios to estimate a chronic

value

, which involves having acceptable chronic toxicity data for the following:

 at least one fish

 at least one invertebrate

 at least one chronic test being from an a cutely

- sensitive species

However, since acceptable chronic values were available for ammonia for all eight

minimum data requirements, the chronic criteri on was derived following the same genus

level sensitivity distribution (SD) approach used to calculate the acute criterion (see 1985

Guidelines for additional detail)

.

 The Guidel ines also require a t least one acceptable test with a freshwater alga or vascular

plant . If plants are among the aquatic organisms most sensitive to the materia l, results of

a plant in another phylum should also be available. The data available on the toxicity of

ammonia to freshwater plants indicate that plants are approximately two orders of

magnitude less sensitive than the aquatic animals tested. Therefore, plant endpoints were

not used in criteria derivation.



# Measures of Effect

Each assessment endpoint requires one or more “measures of ecological effect

, ” which

are defined as changes in the attributes of an assessment endpoint itself or changes in a surrogate

entity or attribute in response to chemical exposure. Ecological effect data are used as measures

of direct and indirect e ffects to biological rece ptors. The measures of effect selected represent

the growth, reproduction, and survival of the organisms

.

The amount of toxicity testing data available for any given pollutant varies significantly,

depending primarily on whether any major environmental issues are raised due to interpretation

## 11

of those data

. A n in

- depth evaluation of available data is performed by EPA t o determine test

acceptability.

Acute measures of effect

Acute measures of effect used for organisms in this document are the LC 50 

## and EC 50



## . LC

stands for “Lethal Concentration” and the LC 50 

## is the concentration of a chemical that is

estimated to kill 50% of the test organisms. EC stands for “Effective Concentration” and the

EC 50 

## is the concentration of a chemical that is estimated to produce a specific effect in 50% of

the test organisms.

As part of the evaluation of new acute data for ammonia, studies submitted using

glochidia, the larval life stage of freshwater mussels in the family Unionidae, were reviewed for

acceptability f

or use in the ammonia criteria development. In 2006 a new ASTM method was

published for toxicity tests with glochidia . H owever, at the time of the 2009 draft revised criteria

for ammonia, EPA and external peer reviewers were concerned that information wa s unavailable

to determine whether the tests with glochidia were ecologically relevant. Specifically, the

appropriate duration of the tests (24, 48, or 96 hrs) was uncertain because it was unclear how the

tests of various durations related to the viabilit y of this short parasitic life stage and its ability to

successfully infect a fish host upon encountering the appropriate fish species. Since that time,

studies by Bringolf et al. (2013) have result ed in the recommendation of a maximum test

duration of 24 hours for glochidia corresponding with the ecologically relevant endpoint of

infectivity for this parasitic life stage. EPA agreed with this recommendation and decided to

include glochidia test s in the criterion dataset for test data with duration s of up to 24 h ou rs with

survival of glochidia at the end of 24 h ou rs of at least 90% in the control treatment

. In addition

,

t o account for species of mussels whose glochidia might not be expected to be viable at 24 hours

(i.e.

, potentially mantle lure strategis ts), EPA examined available test s with glochidia that were

conducted for 24 hours that included testing for viability at 6, 12, and 18 hours. I f the viability

was less than 90% at 24 hours in the control animals , then the next longest duration less than 24

h ours that had at least 90% survival in the control, was considered acceptable for use in deriving

the ammonia criteria

.

## 12

Chronic measures of effect

C hronic measures of effect are EC 20

, 

## N OEC, LOEC , and MATC

. EC 20 

## values we re used

to estimate a low lev el of effect observed in chronic datasets that are available for ammonia (see

U.S. EPA 1999). EC 20 

## is the concentration of a chem ical that is estimated to result in a 2 0

percent effect in a chronic endpoint (e.g., growth, reproduction, and survival ) of th e test

organisms

.

The NOEC ( i.e.

, “No

- Observed

- Effect

- Concentration”) is the highest test concentration

at which none of the observed effects a re statistically different from the control. The LOEC ( i.e.

,

“Lowest

- Observed

- Effect

- Concentration”) is the lowest test concentration at which observed

effects a re found to be statistically different from the control. The MATC is the calculated

geometric mean of the NOEC and LOEC.

For life

- cycle (LC) and partial life

- cycle (PLC) tests, the toxicological variables used in

regression analyses were survival, embryo production, and embryo hatchability. For early life

stage (ELS) tests with fishes, the endpoints used were embryo hatchability, fry/larval survival,

and fry/larval growth . I f ammonia reduced both survival and growth, the product of these

variables (biomass) was analyzed (when possible), rather than analyzing them separately. For

other acceptable chronic and related (e.g., 28

- day juvenile or adult) tests, the toxicological

endpoints analyzed w ere survival, reproduction, hatchability, or growth as appropriate

.

R egression analysis was used, both to demonstrate that a concentration

- effect relationship

was present, and to estimate chronic value s at a consisten t level of effect. E stimates of effect

concentrations can generally be made with precision for a 50 percent reduction in response

(EC 50 

## ) , but at low percent reductions such precision is decreased. A major reduction , such as 50

percent, is not consistent with the intent of establishing chronic criteria to protect the population

f r om long

- term effects . In contrast, a concentration that cause s a low level of reduction in

response, such as an EC 5 

## or EC 10 

## , is rarely statistically significantly different from the control

treatment. EPA selected EC 20 

## values to be used to estimate a low level of effect that would be

statistically different from control effects, yet not so severe as to be expected to cause chronic

impacts at the popul ation level (see U.S. EPA 1999)

. For calculation of the chronic criterion

, the

EC 20 

## point estimate was s e lected for use over a NOEC or LOEC as the measure of effect to use

,

as NOECs and LOECs are highly dependent on test concentrations selected . Furtherm ore, point

estimates provide additional information that is difficult to determine using NOEC and LOEC

## 13

effect measures, such as a measure of effect level across the range of tested concentration s , and

the confidence intervals around those measures of effec t

.

T he typical assessment endpoints for aquatic life criteria are based on unacceptable

effects on growth, reproduction, or survival of the assessed taxa

. The se measures of effect on

toxicological endpoints of consequence to populations are provided by results from the acute and

chronic toxicity tests with aquatic plants and animals

. The toxicity values ( i.e., measures of

effect expressed as genus means) are used in the genus sensitivity distribution of the aquatic

community to derive the aquatic life c riteria. Endpoints used in this assessment are listed in

Table 1

.

Table 1 . Summary of Assessment Endpoints and Measures of Effect Used in Criteria

Derivation for Ammonia

.

Assessment Endpoint s for the Aquatic

Community Measures of Effect

Survival

, growth, and reproduction of

freshwater fish , other freshwater vertebrates,

and invertebrates For acute effects: LC 50 

## or EC 50



## For chronic effects: EC 20 

## , NOEC and LOEC,

calculated MATC

Maintenance and growth of aquatic plants

from standing crop or biomass Not relevant for ammonia because plants are

substantially less sensitive than animals

MATC = maximum ac ceptable toxicant concentration ( geometric mean of NOEC and LOEC)

NOEC = No observed effect concentration

LOEC = Lowest observed effect concentration

LC 50 

### = Lethal concentration to 50% of the test population

EC 50 

### /EC 20 

### = Effect concentration to 50/20% of the test population



## Chronic a veraging p eriod of 30 d ays

T he 30

- day averaging period for chronic effects has been retained from the 1999 chronic

criterion, as is the restriction that the highest 4

- day average within the 30 days may be no greater

than 2.5 times the chronic concentration ( CCC ) more than once every three years on average

.

This is based on analysis of chronic data fr om fathead minnow laboratory tests of different

exposure durations and starting with different age test organisms as summarize d below and

described in greater detail in the 1999 ammonia criteria update.

The 1985 ammonia criteria document specified a CCC averaging period of 4 days as

recommended in the 1985 Guidelines ( Stephan et al. 1985 ), except that an averaging period of 30

days could be used when exposure concentrations were shown to have "limited variability". F or

## 14

ammonia

, the toxicity data on the fathead minnow demonstrate how long the averaging period

should be when concentrations have limited variability , and what restriction applies in terms of

the maximum concentration that can be reached and for how long within that averaging period

.

Based on 7

- day tests, EC 20 

## s of 29.34 and 24.88 mg TAN/L were calculated from the data of

Willingham (1987) , adjusted to pH 7. C hronic values of 20.32 mg TAN/L at pH 7 and 20.99 mg

TAN/L similarly adjusted to pH 7 were reported by Camp Dresser and McKee (1997). T he

geometric m ean of the four values is 2 3.62 mg TAN/L

. This is approximately 2.5 times the

geometric mean EC 20 

## (i.e., 9.396 mg TAN/L at pH 7) for the 30

- day early life

- stage tests

conducted on the same species by Swigert and Spacie (1983) and Mayes et al. (1986)

, [ see also

Appendix B ]

.

Thus, in the 1999 crite ri a document, EPA determined that b ecause the mean chronic

value from the shorter 7

- day toxicity tests with slightly older (< 1 day old) fish is substantially

higher than the mean chronic value from the longer 30

- day ELS tests initiated with newly

fertilized embryos, the CCC averaging period under this “limited variability” can be 30 days, as

long as excursions above the CCC are restricted sufficiently to not exceed the mean chronic

value from the 7

- day tests

. As indicated in the 1999 AWQC document, a more rigorous

definition of this excursion restriction is not possible with the data available, especially because

the information is not available concerning the effects to fish or other animals of variations in

ammonia concentration within a 7

- day test period. It is useful , however, to base the excursion

restriction on a 4

- day period, because this period is the default that already has to be considered

in calculations of water quality

- based effluent limits, a nd because it provides a substantial

limitation of variability relative to the 7

- day chronic values

. While it may be uncertain how

much higher than the CCC the 4

- day average can be, based on the fathead minnow test results

summarized above, 2.5

- fold high er concentrations should be acceptable. O ther data and

justification support ing the use of a longer averaging period for ammonia and the excursion

restriction is provided in the 1999 AWQC document under Chronic Averaging Period (page 81)

.

Ammonia t oxicit y d ata f ulfilling m inimum d ata r equirements

Table 2 provides a summary of the number of toxicity data currently available for gen era

and species that fulfill the 1985 Guidelines minimum requirements for calculation of acute and

chronic criteria for freshwater species exposed to ammonia

.

## 15

T able 2

. 1985 Guidelines Minimum Data Requirements Summary Table Reflecting the

Number of S pecies and G enus L evel Mean Values Represented in the Acute and Chronic

Toxicity Datasets for A mmonia in F reshwater

. Genu s Mean

Acute Value

( GMAV) Species Mean

Acute Value

( 

## SMAV) Genus Mean

Chronic Value

( 

## GMCV) Species Mean

Chronic Value

( 

## SMCV)

Freshwater

F amily Salmonidae in the

class Osteichthyes 4 11 1 3

S econd family in the class

Osteichthyes, preferably a

commercially or

recreationally important

warmwater species 2 2 3 3 6 7

T hird family in the phylum

Chordata (may be in the class

Osteichthye s or may be an

amphibian, etc.) 3 4 1 1

Planktonic Crustacean 4 6 2 3

Benthic Crustacean 6 8 1 1

Insect 9 11 1 1

F amily in a phylum other than

Arthropoda or Chordata ( e.g

., Rotifera, Annelida, or

Mollusca) 1 7 2 3 4 5

F amily in any order of insect

or any phylum not already

represented 4 4 1 a 

## 1 a



## Total 6 9 100 1 7 2 2

a 

### In the absence of other chronic data to fulfill this MDR for another phylum not already represented in the chronic

dataset

, the acute data for species within the phylum Annelida were used to calculate a surrogate chronic value, by

applying a geometric mean ACR from the available invertebrate ACRs.



## Since the data available regarding the toxicity of ammonia to freshwater phytoplankton

and vascular plants reported in the 1985 AWQC document indicate that aquatic plants appear to

be two orders of magnitude less sensitive than the aquatic animals tested, it is assumed that any

ammonia criterion appropriate for the protection of freshwater aquatic animals will also be

protective of aquatic vegetation ( U.S. EPA 1985, 1999, 2009). The greater tolerance of these

taxa to ammonia is due in part to the fact that ammonia is a readily available and energy

- efficient

source of nitrogen for plants; although ammonia can be toxic when present at high

concentrations. For example, the experimental data concerning the toxicity of ammonia to

## 16

freshwater phytoplankton show n egative effects occurring in the green alga, Scenedesmus

obliquus , ranging from approximately 26.88 to 70.14 mg TAN/L with regards to oxygen

evolution and reduction in carbon dioxide photoassimilation (Abeliovich and Azov 1976).

Additionally , ammonia caus ed growth inhibition and cell death of the green alga, Chlorella

vulgaris , at concentrations ranging from 326 to 1,330 mg TAN/L (Przytocka

- Jusiak 1976) ; and

for another algal species, Ochromonas sociabilis , a concentration of 256 mg TAN/L was

algicidal whi le a concentration of approximately half that (128 mg TAN/L ) reduced population

development (assuming pH 6.5 and 30  ̊ C; see Bretthauer 1978). Furthermore, Champ et al.

(1973) investigated the effects of treating a Texas pond with a mean ammonia concentrati on of

25.6 mg/L NH 3 

## (unionized ammonia) for two weeks. A diverse population of dinoflagellates,

diatoms, desmids, and blue

- green algae had been reduced by 95% at the end of the experiment.

At the same time, the pond was virtually eradicated of all rooted aquatic vegetation. Compared

to the 2013 chronic criterion magnitude of 1.9 mg TAN/L, the results from these plant tests,

which are considered as chronic effects according to the 1985 Guidelines, indicate that the 2013

CCC for ammonia will be protecti ve of aquatic plants.

M uch of the early work concerning the response of freshwater vegetation to high

ammonia concentrations is not quantitative or the result of research exploring the possible use of

ammonia as an aquatic herbicide ( U.S. EPA 1985) . T here is no new evidence to suggest that

freshwater phytoplankton and vascular species are more sensitive to ammonia than invertebrates

or fish. Until such a time as those data are produced, EPA will continue to assume that any

ammonia criterion appropriate for t he protection of freshwater aquatic animals will also be

protective of aquatic vegetation.



# Conceptual Model

A c onceptual model consist s of a written description and diagram (U.S. EPA 1998) that

illustrate s the relationships between human activities, stressors, and ecological effects on

assessment endpoints. The conceptual model links exposure characteristics with the ecological

endpoints important for management goals. Under the CWA, these management goal s are

established by states and tribes as designated uses of waters of the United States (for example,

aquatic life support). In deriving aquatic life criteria, EPA is developing acceptable thresholds

## 17

for pollutants that, if not exceeded, are expected to protect designated uses. A state and/or tribe

may implement these criteria by adopting them into their respective water quality standards.

Conceptual d iagram

Environmental exposure to ammonia, while ultimately determined by various site

specific conditio ns and processes, occurs from human activities related to agricultur al practices,

urbanization and industrial processes

, or from natural sources . Point and non

- point sources

contribute to elevated concentrations in ambient surface water. The environmenta l fate

properties of ammonia indicate that direct discharge, runoff, groundwater transport , and

atmospheric deposition represent the pathways of greatest transport to the ambient surface waters

which serve as habitat for aquatic organisms. These sources a nd transport mechanisms are

depicted in the conceptual model below (Figure 2 ). The model also depicts exposure pathways

for biological receptors of concern (e.g., aquatic animals) and the potential attribute changes (i.e.,

effects such as reduced survival, growth and reproduction) in the ecological receptors due to

ammonia exposure.

The conceptual model provides a broad overview of how aquatic organisms can

potentially be exposed to ammonia. Transport mechanisms and exposure pathways are not

quantitatively considered in the derivation of aquatic life criteria

, which are effects assessments

,

not risk assessments . Derivation of criteria focuses on effects on survival, growth and

reproduction of aquatic organisms. However, the pathways, receptors, and attrib ute changes

depicted in Figure 2 may be helpful for states and tribes as they adopt criteria into standards and

need to evaluate potential exposure pathways affecting designated uses.

## 18

Figure 2 : Conceptual M odel for A mmonia E ffects on A quatic A nimals

. ( Available at: http://www.epa.gov/caddis/ssr_amm_int.html )

## 19



# Analysis Plan

During development of CWA sec tion 304(a) criteria , EPA assembles all available

toxicity test data and considers which data are relevant that also meet data quality accepta nce

standards for a ll genera

. Where data allow

, two to four criteri on values are developed (acute and

chronic freshwater , acute and chronic saltwater ). If plants are the most sensitive relative to

vertebrates and invertebrates, plant criteria are developed. T his criteria update document is

specific to ammonia in fresh water, and thus, only two criterion values ( freshwater acute and

chronic) are derived in this document. Available data indicate f reshwater plants are not more

sensitive to ammonia than freshwater animals , thus, plant criteria are not developed

. Fi nally,

ammonia does not bioaccumulate in aquatic animals, thus, final tissue values are not d eveloped.

The se criteria are based on a se nsitivity distribution (SD) comprised of ranked genus

mean acute values (GMAVs), calculated from combined species mean acute values (SMAVs

within each genus ) for acceptable data . SMAVs are calculated using the geo metric mean for all

acceptable measures of effect based on the results of toxicity tests within a given species (e.g.

, all

EC 50 

## s from acceptable acute tests for Daphnia magna ). GMAVs are then calculated using the

geometric means of all SMAVs within a give n genus (e.g.

, all SMAVs for genus Daphnia , such

as Daphnia pulex, Daphnia magna ). I f only one SMAV is available for a genus, then the

GMAV is represented by that value. GMAVs are then rank

- ordered by sensitivity from most

sensitive to least sensitive. The final acute value (FAV) is determined by regression analysis

using a log

- tria ng ular fit based on the four most sensitive genera (reflected as GMAVs) in the

data set to interpolate or extrapolate (as appropriate) to the 5 th 

## percentile of the distribution

represented by the tested genera

. If there are 59 or more GMAVs

, as is the case with ammonia

,

the four GMAVs closest to the 5 th 

## percentile of the distribution are used to calculate the FAV

.

The acute criterion magnitude is the FAV divided by two, in order to provide an acute criterion

magnitude protective of nearly all individuals in 95% of all genera, since the effect endpoint is a

50 th 

## percentile effect (e.g., LC 50 

## or EC 50 

## ) (see 1985 Guidelines , Section XI. Criterion

, B

. )

.

Although the aquatic life criteria derivation process relies on selected toxicity endpoints

from the sensitive species tested, it does not necessarily mean that the selected toxicity endpoints

reflect the sensitivity of the most sensitive species existing in a given environment. The intent of

the eight minimum data requirements is to serve as a sample representative of the aquatic

community. These minimum data requirements represent different ecological, trophic,

## 20

taxonomic and functional differences observed in the natural aquatic ecosystem. The use of the

four most sensitive genera to determine the final criterion value is a censored statistical approach

than improves estimation of the lower tail (most s ensitive) of the distribution when the shape of

the overall distribution, particularly in the less sensitive part of the distribution, is uncertain.

The chronic criterion may be determined by one of two methods. If all eight minimum data

requirements are met with acceptable chronic test data (as is the case with ammonia) , then the

chronic criterion is derived using the same method used for the acute criterion. Genus Mean

Chronic Values (GMCVs) are derived from available Species Mean Chronic Values (SMCVs)

and are then rank

- ordered from least to most sensitive, and the Final Chronic Value (FCV) is

calculated based on regression analysis of a c ensored distribution using the four most sensitive

GMCVs, similar to calculation of the FAV. Unlike the FAV, howeve r, the FCV directly serves

as the basis for the chronic criterion without further adjustment because the endpoint measured

represents a low level (e.g.

, EC 20 

## or NOEC ) of effect (see 1985 Guidelines)

.

In addition, whenever adequately justified, a s tate can develop a site

- specific criterion in

lieu of the use of a national recommended criterion (U.S. EPA 1983)

. The site

- specific criterion

may include not only site

- specific criterion concentrations, but also site

- specific durations o r

averaging periods

, site

- specific frequencies of allowed excursions

, and representative species

present at a given site, where supported by sound science (U.S. EPA 1991). T he Revised

Deletion Process for the Site

- Specific Recalculation Procedure for Aquatic Life Criteria ( U.S

.

EPA 2013 ) provides guidance on revising the taxonomic composition of the toxicity data set

used for the sensitivity distribution upon which a site

- specific criterion is based, in order to better

reflect the assemblage of organisms that resides at the site

. For more information on criteria

derivation, see:

http://water.epa.gov/scitech/swguidance/waterquality/standards/upload/2009_01_13 _criteria_8

5guidelines.pdf

.

The criteria presented are the A gency’s best estimate of maximum ambient

concentrations of ammonia to protect most freshwater aquatic organisms from unacceptable

short

- or long

- term effects. Results of intermediate calculations such as Species Mean Acute

Values ( see in Appendix A ) and chronic values ( see in Appendix B ) are specified to four

significant figures to prevent round ing error in subsequent calculations

, not to reflect the

precision of the value. All of the ammonia acu te values (LC 50 

## s and EC 50 

## s) in Appendix A of this

## 21

document were converted to TAN acute values using the reported temperatures and pHs as

describe d using an example in Appendix D (Conversion of Acute Results of Toxicity Tests)

.

Similarly, all of the ammoni a chronic values (EC 20 

## s) in Appendix B were converted to TAN

chronic v alues as described in Appendix E (Conversion of Chronic Results of Toxicity Tests).



# EFFECTS ANALYSES FOR FRESHWATER AQUATIC ORGANISMS

The acute and chronic ammonia toxicity data used here to update the acute and chronic

criteria for ammonia (freshwater) were collected via literature search es of EPA’s ECOTOX

database, EPA’s Ambient Aquatic Life Water Quality Criteria for Ammonia (U.S. EPA 1985,

1998, 1999), data provided by the U.S. Fish and Wildlife Service and the National Oceanic and

Atmospheric Administration (NOAA) National Marine Fisheries Service (collectively known as

the Services), and EPA regional and field offices. Relevant pap ers were identified, by title and

abstract, and their data s creened according to data quality criteria described in the 1985

Guidelines. All available, reliable acute and chronic toxicity values published since 1985 were

incorporated into the appropriate ammonia AWQC tables and used to recalculate the CMC and

the CCC, as outlined in detail in the 1985 Guidelines. The most recent literature search covered

the period from 1985 through October 2012

.



# Acute Toxicity to Aquatic Animals

All available data relat ing to the acute effects of ammonia on aquatic animals were

considered i n deriving the ammonia criteria and were subjected to a data quality review per the

1985 Guidelines

. The acute effects concentrations are all normalized to pH 7.0 (for all

organisms) and temperature 20 ° C (for invertebrates) as indicated via the equations provided in

Appendix D

. The pH and temperature conditions to which these data are normalized we re

deemed to be generally representative of ambient surface water

. Data that were suitable for the

derivation of a freshwater FAV are presented in Appendix A

.

The GMAVs ranked according to sensitivity, as well as the new (2013) and previous

(1999) acute criterion value s (CMCs), are shown in Figure 3 . The GMAVs represent LC 50 

## s or

EC 50 

## s, whereas the CMC (the FAV/2) values represent concentrations that are expected to be

## 22

lethal to less than 50% of the individuals in either the fifth percentile genus

, or

, a sensitive

commercially or recreationally important species (e.g., adult rainbow trou t).

For this 2013 AWQC document, results from acute toxicity tests that met test

acceptability and quality (according to the 1985 Guidelines) were available for 44 species of fish,

52 species of invertebrates and four species of amphibians . This data includes ammonia toxicity

test data on 52 new species of aquatic animals not previously included in the 1999 acute criterion

dataset

. There are now 6 9 genera represented in the freshwater acute toxicity dataset for

ammonia

, and o f the 6 9 genera ( represent ed in Appendix A and listed according to sensitivity in

Table 3 ) , approximately half are invertebrates. The acute dataset more than fulfills the eight

minimum data requirements outlined in the 1985 Guidelines with between three and 22 genera

represented f

or each taxa category specified (see Table 2 above). The acute criterion dataset now

includes 12 species of aquatic animals Federally

- listed as threatened, endangered or species of

concern. Freshwater invertebrates in the Phylum Mollusca, particularly fr eshwater mussels in

the family Unionidae , freshwater clams

, and some non

- pulmonate snails, are the predominant

group of aquatic organisms ranked in the lowest quartile . T he four most acutely sensitive genera

are all freshwater bivalve mussels (Table 3)

. GMAVs for freshwater mollusk s in general

, are

now among the most influential in th e 2013 acute criterion dataset

.

D ata for glochidia and juvenile life stages of freshwater unionid mussels were evaluated

for acceptability based on the 1985 Guidelines, the approved ASTM protocol for toxicity testing

with these life stages of unionid mussels (ASTM 2006), and recent studies on the most

ecological ly relevant toxicological endpoint (s) and exposure duration (s) for glochidia tests by

Bringolf et al . (201 3 ). The acute unionid mussel dataset for ammonia now includes acceptable

data for 11 genera, totaling 16 species of freshwater mussels, as well as two sensitive species of

non

- pulmonate snails. Of these

, four of the 18 mollusk species included in 2 013 acute dataset

are Federally

- listed as threatened or endangered (as identified in Table 3 ).

Nearly all states in the continental United States have freshwater unionid mussel fauna in

at least some of their waters (Abell et al. 2000 ; Williams et al. 19 93 ; Williams and Neves 1995).

While the number of freshwater unionid mussel species is less and the distribution is sparse in

the dry western states, even New Mexico and Arizona have at least one native mussel species

(Williams et al. 1993). Moreover, ap proximately one

- quarter of nearl y 300 freshwater unionid

mussel taxa in the USA are Federally

- listed as endangered, threatened or of special concern. In

## 23

addition, non

- pulmonate snails are relatively ubiquitous compared to mussels and of the 650

freshwater snail species

, 25 species are Federally

- listed. Every state in the continental U.S. has

at least one family of non

- pulmonate snail in at least some of their waters. Thus, considering

that f reshwater unionid mussels are among some of the most sensitive g enera in the dataset , and

that all states have at least one freshwater unionid mussel or bivalve mollusk

, or non

- pulmonate

snail species, another relatively sensitive mollusk group, native or present in at least some of

their waters, EPA is recommend ing a single national acute criterion to be applied to all waters

rather than different criteria based on the presence or absence of mussels

.

The most sensitive fish SMAV is for mountain whitefish, Prosopium williamsoni (SMAV

of 51.93 mg TAN/L ), representing o ne of the four genera of salmonids in the acute dataset,

followed by the second most sensitive fish

, the Lost River sucker (SMAV of 56.62 mg TAN/L )

,

which is an endangered species (Table 3)

. The mountain whitefish GMAV is ranked eighth most

sensitive afte r seven more sensitive GMAVs for freshwater mussel species, thus

, salmonids

should be adequately protected by the new acute criteri on

. The next most sensitive salmonid

genus is On corhynchus , represented by data for six different species, three of which are

threatened or endangered, with SMAVs ranging from 78.92 mg TAN/L for Cutthroat trout, O.

clarkii

, to 180.7 mg TAN/L for pink salmon, O. gorbuscha

. T he GMAV for Oncorhynchus

(99.15 mg TAN/L ) is ranked #25 in acute sensitivity rank at pH 7 and temperature 20 ° 

## C (Table

3)

.

T he four lowest GMAVs in this 201 3 ammonia AWQC update are for invertebrate

species (specifically, freshwater bivalve mollus ks dominated by mussels in the f amily

Unionidae). Because the most sensitive GMAVs are all represented by invertebrate species, the

CMC is both pH

- dependent, in accordance with the acute pH

- toxicity relationship for all aquatic

organisms , and temperature

- dependent, due to the invertebrate acute

- temperature relationship.

## 24

Figure 3

. Ranked Freshwater Genus Mean Acute Values (GMAVs) with Criterion

Maximum Concentrations (CMCs).

Summaries of studies used in acute crite r ion determination

Presented in this section are brief summaries of the results of acute toxicity tests that

meet the data quality acceptability criteria and that are used directly for deriving the FAV (i.e.,

serve as the basis for the SMAV or GMAV of one of the most sensitive genera). A s per the 1985

Guidelines, whenever there are 59 or more GMAVs in the acute criteria dataset, the FAV is

calculated using the four GMAVs closest to the 5th percentile of the distribution

.

The four species and associated endpoints (SMAV or GMAV) used in calculating the

acute criteri on (sensitivity rank 2

- 5) are ranked below from most to least sensitive:

## 25

2. Lasmigona subviridis

, Green Floater ( G MAV = 23.41 mg TAN/L )

3. Epioblasma capsaeformis

, Oyster mussel ( G MAV = 31.14 mg TAN/L )

4. Villosa iris , Rainbow Mussel (GMAV= 34.23 mg TAN/L )

5. Lampsilis sp. (GMAV

= 46.63 mg TAN /L )

T he most sensitive species Venustaconcha ellipsiformis (SMAV=23.12 mg TAN/L ) is not

included in the criteria numeric calculation , because it falls below the 5 th 

## percentile in sensitivity

in the distribution of 69 genera included in the dataset

.

Summar ies are provided on the basis of individual species or genera (in cases where more

than one species is included in the calculation of the GMAV). All values are provided in terms

of total ammonia nitrogen (TAN) , either as reported by the authors or as converted from the

reported values for unionized ammonia, pH, and temperature (using the speciation relationship)

applied in the 1999 AWQC document (i.e., Emerson et al. 1975). I n the special cases where the

resul t of a test is considered an upper limit on an acute value, the value is ascribed a greater than

( “>” ) sign indicating as much.

Lasmigona subviridis (green floater)

The GMAV /SMAV for the green floater , a freshwater bivalve mollusk, of 23.41 mg

TAN/L is b ased on the geometric mean of three 96

- hr EC 50 

## s from tests using less than two

month old juveniles as reported in Black (2001). Test solutions were renewed after 48 hours.

The mean pH and test temperature for two of the tests was 7.73 and 24 ° C, and for t he third, 7.92

and 24.8 ° C. Control survival exceeded 90 percent in all three tests. The reported EC 50 

## s at test

temperature and pH expressed on the basis of TAN were 6.613, 6.613 and 3.969 mg TAN/L

,

respectively. Adjusted to pH 7 and 20 ° C , the EC 50 

## s are 24.24, 24.24 and 21.84 mg TAN/L

,

respectively ( Appendix A ). The GMAV for juvenile green floaters of 23.41 mg TAN/L

represents the second lowest in the acute dataset, and the lowest of the four GMAVs used to

calculate the FAV (Table 3)

.

Epioblasma capsaef

ormis (oyster mussel)

The GMAV /SMAV for the endangered oyster mussel (31.14 mg TAN/L ) is the third

lowest in the acute dataset (Table 3 ), and is based on the geometric mean of a 96

- hr EC 50 

## from a

## 26

renewal test using less than five

- day old juveniles, and two 6

- hr EC 50 

## s from static tests

conducted with two

- hour old glochidia (Wang et al. 2007b). The mean pH and test temperature

for all three tests was 8.5 and 20 ° C. Control survival exceeded 90 percent in all tests. The

estimated measured EC 50 

## for juvenile oyster mussels at test temperature and pH was 4.760 mg

TAN/L , after adjusting the reported nominal E C 50 

## by m ultiplying by a factor of 0.835 (i.e.,

measured total ammonia concentrations were 83.5 percent of nominal concentrations for 96 hour

j uvenile exposures) . T he reported EC 50 

## s for glochidia were 3.4 and 5.0 mg TAN/L , respectively

(no further adjustment necessary). These EC 50 

## s normalized to pH 7 and 20 ° C are 53.63, 17.81

and 31.61 mg TAN/L

, for the two glochidia and juveniles respectively ( Appendix A ).

Villosa iris (rainbow mussel)

Ten EC 50 

## s from several studies (Goudreau et al. 1993; Scheller 1997; Mummert et al.

2003 ; Wang et al. 2007b ) using two different life stages (glochidia and juvenile) and range of

ages within each life

- stage were used to calculate the GMAV/SMAV for rainbow mussel

(Appendix A). All tests were either static or static renewal where concentrations were measured.

The GMAV of 34.23 mg TAN/L is the fourth lowest in the acute dataset (Table 3), and is

composed of indivi dual EC 50 

## values (expressed as TAN and normalized to pH 7 and 20 ° C)

ranging from 12.62 to 99.28 mg TAN/L (Appendix A). The difference in pH and test

temperature among the 10 different tests ranged from 7.29 to 8.40 and 12.6 to 25.0 ° C,

respectively. Contr ol survival exceeded 90 percent in all tests regardless of life

- stage tested.

The glochidia were not substantially more sensitive than the juveniles (less than a factor of 2

difference)

.

Mussels in Genus Lampsilis

Freshwater unionid mussels within the Genus Lampsilis represent the most widely tested

genus to date. The GMAV of 4 6.63 mg TAN/L reflects the geometric mean of SMAVs for six

species, two ( Lampsilis abrupta and L. higginsii ) which are endangered and a thir d ( L.

rafinesqueana ) that is a Federal species of concern ( Table 3 ). The SMAVs for this genus range

from 26.03 mg TAN/L ( L. abrupta ) to 69.97 mg TAN/L ( L. rafinesqueana ), and are composed

of anywhere from one ( L. abrupta ) to four teen ( L. siliquoidea ) indi vidual EC 50 

## s ( Appendix A ).

T he range of EC 50 

## s used to calculate the FAV, normalized to pH 7 and 20 ° C across all species of

## 27

Lampsilis is from 24.30 to 160

. 5 mg TAN/L ( see Appendix A ). The GMAV for Lampsilis is the

fifth most sensitive in the acute dataset , and the highest of the four GMAVs used to calculate the

FAV (Table 3)

. Both glochidia and juvenile data were available for three of the six Lampsilis

species , showing an inconsistent pattern of relative sensitivity.

Table 3. Ranked Genus Mean Acute Values

.

Table 3. Ranked Genus Mean Acute Values

Rank GMAV

( mg TAN/L ) Species SMAV

( mg TAN/L )

69 2515 Insect,

Erythromma najas 2515

68 994.5 Caddisfly,

Philarctus quaeris 994.5

67 735.9 Beetle,

Stenelmis sexlineata 735.9

66 686.2 Crayfish,

Orconectes immunis 1550

Crayfish,

Orconectes nais 303.8

65 681.8 Midge,

Chironomus riparius 1029

Midge,

Chironomus tentans 451.8

64 442.4 Mayfly,

Drunella grandis 442.4

63 387.0 Aquatic sowbug,

Caecidotea racovitzai 387.0

62 378.2 Isopod,

Asellus aquaticus 378.2

61 281.5 Threespine stickleback,

Gasterosteus aculeatus 281.5

60 246.5 Mayfly,

Callibaetis skokianus 364.6

Mayfly,

Callibaetis sp. 166.7

59 233.0 Dragonfly,

Pachydiplax longipennis 233 .0

58 222.2 Mottled sculpin,

Cottus bairdii 222.2

57 219.3 Western mosquitofish,

Gambusia affinis 219.3

56 218.7 Oligochaete worm,

Lumbriculus variegatus 218.7

55 216.5 Tubificid worm,

Tubifex tubifex 216.5

54 211.6 Marsh ramshorn snail,

Planorbella trivolvis 211.6

## 28 Table 3. Ranked Genus Mean Acute Values

Rank GMAV

( mg TAN/L ) Species SMAV

( mg TAN/L )

53 192.6 Scud,

Hyalella azteca 192.6

52 192.4 Stonefly,

Skwala a mericana 192.4

51 185.2 Mozambique tilapia,

Oreochromis mossambicus 185.2

50 181.8 Amphipod,

Crangonyx pseudogracilis 270.5

Amphipod,

Crangonyx sp. 122.2

49 170.2 Tubificid worm,

Limnodrilus hoffmeisteri 170.2

48 164.5 Pouch snail,

Physa gyrina 164.5

47 164.0 Damselfly,

Enallagma sp. 164.0

46 162.6 Water flea,

Chydorus sphaericus 162.6

45 159.2 Fathead minnow,

Pimephales promelas 159.2

44 157.8 Brook trout,

Salvelinus fontinalis 156.3

Lake trout,

Salvelinus namaycush 159.3

43 156.7 Shortnose sturgeon,

Acipenser brevirostrum (LS) 156.7

42 146.5 White sucker,

Catostomus commersonii 157.5

Mountain sucker,

Catostomus platyrhynchus 136.2

41 143.9 Water flea,

Ceriodaphnia acanthine 154.3

Water flea,

Ceriodaphnia dubia 134.2

40 142.9 Water flea,

Simocephalus vetulus 142.9

39 142.4 Channel catfish,

Ictalurus punctatus 142.4

38 138.0 Red swamp crayfish,

Procambarus clarkii 138.0

37 136.7 Atlantic salmon,

Salmo salar (LS) 183.3

Brown trout,

Salmo trutta 102.0

36 134.8 White perch,

Morone americana 132.7

White bass,

Morone chrysops 144.0

## 29 Table 3. Ranked Genus Mean Acute Values

Rank GMAV

( mg TAN/L ) Species SMAV

( mg TAN/L )

Striped bass,

Morone saxatilis 246.2

Sunshine bass,

Morone saxatilis x chrysops 70.22

35 125.0 Water flea,

Daphnia magna 157.7

Water flea,

Daphnia pulicaria 99.03

34 122.5 Clawed toad,

Xenopus laevis 122.5

33 119.5 Flatworm,

Dendrocoelum lacteum 119.5

32 117.1 Walleye,

Sander vitreus 117.1

31 115.9 Central stoneroller,

Campostoma anomalum 115.9

30 110.0 Rainbow dace,

Cyprinella lutrensis 196.1

Spotfin shiner,

Cyprinella spiloptera 83.8 0

Steelcolor shiner,

Cyprinella whipplei 80.94

29 109.0 Dwarf wedgemussel,

Alasmidonta heterodon (LS) 109.0

28 109.0 Pink papershell,

Potamilus ohiensis 109.0

27 106.9 Green sunfish,

Lepomis cyanellus 150.8

Pumpkinseed,

Lepomis gibbosus 77.53

Bluegill,

Lepomis macrochirus 104.5

26 106.3 Common carp,

Cyprinus carpio 106.3

25 99.15 Golden trout,

Oncorhynchus aguabonita 112.1

Cutthroat trout,

Oncorhynchus clarkii 78.92

Pink salmon,

Oncorhynchus gorbuscha 180.7

Coho salmon,

Oncorhynchus kisutch (LS) 87.05

Rainbow trout,

Oncorhynchus mykiss (LS) 82.88

Chinook salmon,

Oncorhynchus tshawytscha (LS) 82.39

24 96.72 Topeka shiner,

Notropis topeka (LS) 96.72

## 30 Table 3. Ranked Genus Mean Acute Values

Rank GMAV

( mg TAN/L ) Species SMAV

( mg TAN/L )

23 96.38 Leopard frog,

Rana pipiens 96.38

22 89.36 Long fingernailclam,

Musculium transversum 89.36

21 89.06 Smallmouth bass,

Micropterus dolomieu 150.6

Largemouth bass,

Micropterus salmoides 86.02

Guadalupe bass,

Micropterus treculii 54.52

20 88.62 Great pond snail,

Lymnaea stagnalis 88.62

19 74.66 Guppy,

Poecilia reticulata 74.66

18 74.25 Johnny darter,

Etheostoma nigrum 71.45

Orangethroat darter,

Etheostoma spectabile 77.17

17 72.55 Rio Grande silvery minnow,

Hybognathus amarus 72.55

16 71.56 Spring peeper,

Pseudacris crucifer 61.18

Pacific tree frog,

Pseudacris regilla 83.71

15 71.25 Mucket,

Actinonaias ligamentina 63.89

Pheasantshell,

Actinonaias pectorosa 79.46

14 70.73 Giant floater mussel,

Pyganodon grandis 70.73

13 69.36 Shortnose sucker,

Chasmistes brevirostris 69.36

12 68.54 Pagoda hornsnail,

Pleurocera uncialis 68.54

11 63.02 Golden shiner,

Notemigonus crysoleucas 63.02

10 62.15 Pebblesnail,

Fluminicola sp. 62.15

9 56.62 Lost River sucker,

Deltistes luxatus (LS) 56.62

8 51.93 Mountain whitefish,

Prosopium williamsoni 51.93

7 47.40 Atlantic pigtoe,

Fusconaia masoni 47.40

6 46.93 Pondshell mussel,

Utterbackia imbecillis 46.93

5 46.63 Pink mucket,

Lampsilis abrupta (LS) 26.03

## 31 Table 3. Ranked Genus Mean Acute Values

Rank GMAV

( mg TAN/L ) Species SMAV

( mg TAN/L )

Plain pocketbook,

Lampsilis cardium 50.51

Wavy

- rayed lampmussel,

Lampsilis fasciola 48.11

Higgin's eye,

Lampsilis higginsii (LS) 41.90

Neosho mucket,

Lampsilis rafinesqueana (LS) 69.97

Fatmucket,

Lampsilis siliquoidea 55.42

4 34.23 Rainbow mussel,

Villosa iris 34.23

3 31.14 Oyster mussel,

Epioblasma capsaeformis (LS) 31.14

2 23.41 Green floater,

Lasmigona subviridis 23.41

1 23.12 Ellipse,

Venustaconcha ellipsiformis 23.12

FAV = 33.52

CMC = 1 7

LS = Federally

- listed as threatened or endangered species



# Chronic Toxicity to Freshwater Aquatic Animals

Freshwater c hronic toxicity data that meet the test acceptability and quality

assurance/control criteria are presented in Appendix B

. All tests were conducted with measured

concentrations of ammonia. Ammonia chronic toxicity data are available for 21 species of

fres hwater organisms: ten invertebrate species ( mussels, clam, snail, cladoc erans , daphnid, and

insect) and 11 fish species , including three Federally

- listed salmonid species

. T he chronic

dataset includes data for three freshwater unionid mussel species, one freshwater non

- pulmonate

snail species, and two fish species not included in the 1999 criteria (see Appendix B ). It also

includes an estimate of chronic effects for the Phylum Annelida, to meet the data requirement of

a species in “ a f

amily in any order of insect or any phylum not already represented ,” as described

below

.

Each chronic test was reviewed to determine acceptability based on the dilution water,

control mortality, experimental design, organism loading, etc.

, as consistent with ASTM

standards, including for freshwater mussels via E2455

- 06 (ASTM 2006). The concentration of

## 32

dissolved oxygen was also reviewed to determine acceptability based on the general limits

specified in the 1999 AWQC document. The mean measured dissolved oxygen concentratio n

and the lower limit for dissolved oxygen concentration required to be protective varies based on

taxa group . T he mean dissolved oxygen concentration for toxicity tests should be at least 6.5

mg/L for salmonids

, 6.0 mg/L for invertebrates

, and the lower limit of dissolved oxygen should

be 5.0 mg/L to be protective of both of these group s o f organisms (U.S. EPA 1999).

Based on the determination that the test methodology used was acceptable, the studies

w ere evaluated to determine whether the ammonia cause d a reduction in (a) survival (if over a

period of at least seven days), (b) growth , or (c) reproduction . If the test demonstrated reduction

in any of these toxicological endpoints , the test could be accepted for use in calculating the

chronic value ( CV )

.

A cceptable 28

- day survival tests using juvenile freshwater mussels and juvenile

freshwater snails and growth tests using juvenile freshwater snails we re evaluated for inclusion

in the derivation of the chronic aquatic life criteri on when the test concent ration cause d a

reduction in survival or growth of 20 percent or more of these types of organisms at those life

stages

. Based on evaluation of the individual studies (Wang et al. 2007a ; Wang et al. 2011),

growth data for juvenile mussels was not used in t he derivation of the chronic criterion due to

uncertainty in method of measurement for the growth endpoint (see Effects Characterization for

further discussion).

All chronic data in individual studies w ere analyzed using r egression analysis to

demonstrat e th e presence of a concentration

- effect relationship within the test . For those studies

that demonstrated a concentration

- effect relationship, EPA used regression analysis to estimate

the EC 20



## .

Sixteen GMCVs are presented in Appendix B and ranked according to sensitivity in

Table 4

. The four lowest values were used to calculate the FCV, because values for fewer than

59 genera exist. EPA calculat ed the chronic criterion based on fifth percentile of the GMCVs in

Table 4. The GMCVs for the four most sensitive species are ranked below fro m most to least

sensitive :

1. Lampsilis spp

, Wavy

- rayed lamp mussel and Fatmucket (GMCV= 2.126 mg TAN/L )

2. Villos a iris, Rainbow m ussel (GMCV= 3.501 mg TAN/L )

## 33

3. Lepomis spp., Bluegill and Green s unfish (GMCV= 6.920 mg TAN/L )

4. Musculium transversum, Long fingernail clam ( GMCV= 7.547 mg TAN/L )

T he chronic criterion magnitude is 1. 9 mg TAN/L at 2 0 ° C and pH 7

. The four most

sensitive species are predominantly mollusk s although Lepomis species (bluegill and green

sunfish) compris e the third most sensitive GMCV. Figure 4 shows t he GMCVs ranked

according to sensitivity and shows the 2013 chronic criteria magnitude as well as the 1999

criterion value ( based on fish early life stages) for comparative purposes.

Figure 4 . Ranked Freshwater Genus Mean Chronic Values (GMCVs) with Criterion

Continuous Concentrations (CCCs).

## 34

Summaries of studies used in chronic criterion determination

The following presents a species

- by

- species discussion of freshwater chronic data used in

deriving the chronic crite r ion magnitude for ammonia. All analyses were conducted in terms of

TAN , either as reported by the authors or as converted from the reported values for unionized

ammonia, pH, and temperature ( using the speciation relationship in Emerson et al. 1975). EC 20



## values were adjusted to pH 7 , and for invertebrates, also adjusted to a temperature of 2 0 ° C

.

SMCVs were used when data were available for only one species. When data for more than one

species in a tax on we re available, GMCVs were calculated from the SMCVs. All of the CVs

(EC 20 

## values), SMCVs, and GMCVs derived are tabulated and included in Appendix B

. F or

some of the new chronic data, authors reported EC 20 

## values on the basis of TAN . In such cases

these reported CVs were normalized to pH 7 and 2 0 ° C (temperature normalization for

invertebrates only), and utilized for the analysis. The results of all intermediate calculations such

as ECs, SMCVs and GMCVs are given to four significant figures to prevent round

- off error in

subsequent calculations, not to reflect the precision of the value.

Lampsilis species

Lampsilis fasciola (wavy

- rayed lampmussel)

Wang et al. (2007a) published results of the effect of ammonia on survival and growth of

2

- month old juvenile freshwater unionid mussels. The 28

- day juvenile test was part of a series

of studies designed to refine the methods for conducting acute and chro nic toxicity tests with

early life stages of freshwater mussels. Dissolved oxygen was maintained above 7.0 mg/L

during the 28

- day test. Survival in the control treatment and lowest ammonia test concentration

(0.13 mg TAN/L ) were 100 and 83 percent, respe ctively. Survival decreased to 30 percent at

1.02 mg TAN/L , and zero at 1.98 mg TAN/L

. There was no concentration

- response relationship

for either length at 28 days or change in length after 28 days. Using EPA’s TRAP model (see

Appendix G ), the survival EC 20 

## for this freshwater unionid mussel species is 0. 4272 mg TAN/L

at test temperature (20 ° C) and pH (8.2), or 1. 408 mg TAN/L when adjusted to pH 7 and 2 0 ° C

( Appendix B ).

## 35

Lampsilis siliquoidea (fatmucket)

In a recent study, Wang et al. (2011) evaluated t he influence of substrate on the

sensitivity of two

- month

- old juvenile mussels to ammonia in 28

- day water

- only exposure and

substrate exposure. The methods used were similar to th ose in an earlier study ( Wang et al.

2007a ) except for how the organisms wer e exposed. In this study, the organisms were housed in

a glass tube with a screen bottom that was suspended in a beaker. The authors conducted two

exposure conditions simultaneously for comparison of the water

- only and substrate exposure.

The organisms used in the w ater

- only exposure were simply placed on the screen at the bottom

of the tube . The substrate treatment involved substrate that was screened, eliminating both large

and small particles, with only particles between 300

- 500 microns retained, which is essentially

the grain size of medium sand. A layer of substrate was placed on the s creen and the organisms

were placed on top of the relatively inert substrate. In the substrate treatment, the water actively

flowed past the organisms and through the substrate. Water chemistry was characterized before

and after passing through the subst rate and found not to be substantially altered . Furthermore,

the pH was maintained consistently at approximately pH 8. 25 in overlying water and pore water.

The survival response between the water

- only and substrate treatments was similar with a

reported LO EC of 0.53 mg TAN/L in the water

- only and 0.88 mg TAN/L for the substrate

treatment at the test pH 8. 25 and temperature 2 0 ° C. Mean control survival in both the water

only and substrate treatments was 95% at the end of the 28

- day exposures , which me t

acce ptability requirements

. Dry weight measurements of the mussels increased by 165% in the

water

- only exposure compared to 590% increase in the substrate exposure suggesting that the

presence of the substrate increased food availability, as noted by the auth ors.

Using TRAP threshold sigmoid regression of the survival response results in an EC 20 

## of

0.5957 mg TAN/L for the water

- only and EC 20 

## 0.8988 mg TAN/L for the substrate exposure at

the test pH and temperature, or adjusted to pH 7 and 20 ° 

## C

, chronic value s equivalent to 2.128

and 3.211 mg TAN/L , respectively (Appendix B)

. Based on the apparent improved health of the

test organisms in the substrate exposures

, and the lack of a ny significant a lter ation of water

chemistry in the exposure , the SMCV 3.211 mg T AN/L

, based on survival of juvenile fatmucket

from the substrate exposures is used to calculate the CCC rather than the water

- only exposure.

## 36

The geometric mean of SMCV s for fatmucket and wavy

- rayed lamp mussel of 3.211 and

1.408 mg TAN/L , respectively, results in a GMCV of 2.126 mg TAN/L for the genus Lampsilis

(Table 4).

Villosa iris (rainbow mussel)

The effect of ammonia on survival and growth of th is freshwater unionid mussel species

was also reported in the study by Wang et al. (2007a). Juvenile ( 2

- month

- old) rainbow mussels

were tested via a 28

- day test under similar conditions as described above. Survival was ≥ 98

percent up to the 0.81 mg TAN/L exposure, but fell to 15 percent at 1.67 mg TAN/L and zero

percent at 3.45 and 7.56 mg TAN/L . EPA’s TRAP was used to generate a chronic value for this

species based on survival resulting in E C 20 

## of 1.063 mg TAN/L at test temperature (20 ° C) and

pH (8.2)

– ( Appendix G )

, or 3.501 mg TAN/L a djusted to pH 7.0 . Wang et al. (2007a) elected to

exclude length estimates for concentrations above those where significant survival effects were

measured (or in this case, 1.67 mg TAN/L ). As a result, growth data are available for only three

effect concentrations, even though there was 15% survival at the 1.67 mg TAN/L treatment

level

. Due to the uncertainties in the limited growth data for this test the growth data was not

used in the calculation of the GMCV.

The SMCV and GMCV for this freshwater unionid mussel spe ci es is 3.501 mg TAN/L

when adjusted to pH 7 and 2 0 ° C (Appendix B)

.

Lepomis sp ecies

Lepomis cyanellus (green sunfish)

Reinbold and Pescitelli (1982a) conducted a 31

- day early life

- stage ( ELS ) test that started

with <24

- hour

- old embryos. No information was reported concerning the DO concentration

, but

it averaged 70 to 76 percent of saturation (5.7 to 6.2 mg/L) in a similar test in the same report

with another fish species at about the same temperature. T he weight data were not used in the

calculation of an EC 20 

## because of the greater weight of the fish in test chambers containing fewer

fish, which indicated that weight was density

- dependent. Although overflows resulted in loss of

fish from some chambers, survival was 96 percent in one of the chambers affec ted by overflow,

indicating that the survival data were either adjusted or not affected by the overflows. Survival

by the end of the test was reduced at test concentrations of 6.3 mg TAN/L and above. TRAP

## 37

analysis of the survival data resulted in an EC 20 

## of 5.840 mg TAN/L at pH 8.16 and 25.4 ° 

## C (U.S.

EPA 1999). Adjusted to pH 7, the EC 20 

## is 18.06 mg TAN/L (Appendix B).

McCormick et al. (1984) conducted a 44

- day ELS test starting with <24

- hour

- old

embryos. During this test, no effect was found on percent h atch, but survival and growth were

both reduced at measured test concentrations of 14 mg TAN/L and above. TRAP analysis using

biomass resulted in an EC 20 

## of 5.61 mg TAN/L at pH 7.9 and 22.0 ° 

## C for the test (U.S. EPA

1999). Adjusted to pH 7, the EC 20 

## calcu lated using the data as previously reported in U.S. EPA

(1999) is 11.85 mg TAN/L (Appendix B).

The pH

- adjusted EC 20 

## s of 18.06 mg TAN/L from Reinbold and Pescitelli (1982a) and

11.85 mg TAN/L from McCormick et al. (1984) agree well with one another. I t is possible that

the second value is lower because it was based on survival and growth, whereas the first value

was based only on survival. T he results of the tests were deemed acceptable for use in

calculating a SMCV for the species, which is 14.63 mg TAN/L (Table 4) at pH 7

.

Lepomis macrochirus (bluegill)

Similar to the studies summarized above for L. cyanellus , Smith et al. (1984) conducted a

30

- day ELS test starting with <28

- hour old embryos of L. macrochirus . No information was

reported concerning the DO concentration, but the flow

- rate was kept high during the test. In

this study, the authors found no significant reduction in percent hatch up to a test concentration

of 37 mg TAN/L , but hatched larvae were deformed at this concentration and died within six

days. By the end of the test, both survival and growth were greatly reduced at measured test

concentrations ranging from 3.75 to 18 mg TAN/L . TRAP analysis of biomass resulted in

calculation of an EC 20 

## o f 1.85 mg TAN/L at pH 7.76 and 22.5 ° 

## C ( U.S. EPA 1999). The EC 20



## adjusted to pH 7 is 3.273 mg TAN/L (Appendix B).

The SMCV for the bluegill is 3.273 mg TAN/L , which, when calculated as a geometric

mean with the SMCV of 14.63 mg TAN/L for green sunfish, res ults in a GMCV of 6.920 for the

genus Lepomis (Table 4).

Musculium transversum

Anderson et al. (1978) conducted two 42

- day tests of the effect of ammonia on survival

of field

- collected juvenile clams whose length averaged 2.2 mm. The results of the two t ests

## 38

were similar so the data were pooled for analysis. Survival in the control treatment and low

ammonia concentrations (<5.1 mg TAN/L ) ranged from 79 to 90%, but decreased to zero at 18

mg TAN/L . TRAP analysis of the survival data resulted in a calcula ted EC 20 

## of 5.820 mg

TAN/L at 23.5 ° 

## C and pH 8.15. The EC 20 

## is 22.21 mg TAN/L when adjusted to pH 7 and 20 ° 

## C

(Appendix B).

Sparks and Sandusky (1981) conducted a test similar to Anderson et al. (1978) with field

collected juvenile clams whose average lengt h was 2.1 mm. The test was conducted in the same

laboratory and used test organisms from the same location in the Mississippi River as Anderson

et al. (1978), but employed a feeding regime and food for the test that was deemed by the authors

to be better suited to maintaining the health of fingernail clams during chronic toxicity testing.

Survival in the control treatment was 92% and decreased with increasing concentration of

ammonia to 17% at 18 mg TAN/L . Effects on survival were evident at lower concen trations,

resulting in an EC 20 

## of only 1.23 mg TAN/L at 21.8 ° 

## C and pH 7.80 when calculated using

TRAP. The EC 20 

## adjusted to pH 7 and 20 ° 

## C is 2.565 mg TAN/L (Appendix B).

Although this latter EC 20 

## determined for the test reported by Sparks and Sandusky (1981)

is substantially lower than that obtained by Anderson et al. (1978), the difference is less than a

factor of 10, and thus, the SMCV for this species (at pH 7 and 20 ° 

## C) is the geometric mean of the

two values, or 7.547 mg TAN/L (Table 4).

## 39

Table 4

. Ranked Genus Mean Chronic Values.

Rank GMCV

( mg TAN/L ) Species SMCV

( mg TAN/L )

16 73.74 Stonefly,

Pteronarcella badia 73.74

15 53.75 Water flea,

Ceriodaphnia acanthina 64.1 0

Water flea,

Ceriodaphnia dubia 45.08

14 41.46 Water flea,

Daphnia magna 41.46

13 29.17 Amphipod,

Hyalella azteca 29.17

12 21.36 Channel catfish,

Ictalurus punctatus 21.36

11 20.38 Northern pike,

Esox lucius 20.38

10 16.53 Common carp,

Cyprinus carpio 16.53

9 12.02 Lahontan cutthroat trout,

Oncorhynchus clarkii henshawi

(LS)* 25.83

Rainbow trout,

Oncorhynchus mykiss (LS) 6.663

Sockeye salmon,

Oncorhynchus nerka (LS) 10.09

8 11.62 White sucker,

Catostomus commersonii 11.62

7 11.07 Smallmouth bass,

Micropterus dolomieu 11.07

6 9.187 Fathead minnow,

Pimephales promelas 9.187

5 7.828 Pebblesnail,

Fluminicola sp. 7.828

4 7.547 Long fingernailclam,

Musculium transversum 7.547

3 6.920 Green sunfish,

Lepomis cyanellus 14.63

Bluegill,

Lepomis macrochirus 3.273

2 3.501 Rainbow mussel,

Villosa iris 3.501

1 2.126 Fatmucket,

Lampsilis siliquoidea 3.211

Wavy

- rayed lamp mussel,

Lampsilis fasciola 1.408

FCV = 1.887 mg TA N/L

CCC = 1.9 mg TAN/L

LS = Federally

- listed species as threatened or endangered

LS * = Listed at the subspecies only for specific population s

## 40



# T he National Criteria for Ammonia in Fresh Water

This ammonia criteria update document recommends an acute criterion magnitude of 17

mg TAN/L and a chronic criterion magnitude of 1.9 m g TAN/L at pH 7 and 20 ° C , with the

stipulation that the chronic criterion cannot exceed 4.8 mg TAN/L as a 4

- day average . All

criteria magnitudes are recommended not to be exceeded more than once in three years on

average

.

2013 Final A quatic L ife Criteria for Ammonia

(Magnitude, Frequency, and Duration)

(mg TAN/L)

pH 7.0, T=20 ° C

Acute

(1

- hour average) 17

Chronic

(30

- day rolling average) 1.9*

*Not to exceed 2.5 times the CCC as a 4

- day average within

the 30

- days, i.e. 4.8 mg TAN/L at pH 7 and 20 ° C , more than

once in three years on average.

Criteria frequency: Not to be exceeded more than once in

three years on average.

T he available data for ammonia indicate that, except possibly where an unusually

sensitive species is important at a site, freshwater aquatic life will be protected if these criteria

are met. Tables 5 a and 5b below provide the temperature and pH

- dependent values of the CMC

(acute criterion magnitude ) and Table 6 provides the temperature and pH

- dependent values of

the CCC (chronic criterion magnitude ) based on the following recommended criterion

calcula tions derived for this update.

Acute c riterion c alculat ion s

The one

- hour average concentration of total ammonia nitrogen (in mg TAN/L ) is not to

exceed, more than once every three years on the average, the CMC (acute criterion magnitude )

calculated using the following equation:

## 41 ( ( )

( ( ) ( ( ) 

### ) ) )



## The 2013 CMC equation is predicated on the following:

1. T he lowest GMAV in this criterion update is for an invertebrate species; thus, the CMC is

both pH

- and temperature

- dependent and varies with temperature according to the invertebrate

acute temperature relationship

. The lowest GMAV is 23.12 mg TAN/L for Venustaconcha

(Table 3). The updated CMC ( rounded to 4 significant figures) of 16.76 mg TAN/L at pH 7 and

2 0 ° C is 27 .5 percent lower than this value. The CMC divided by the lowest GMAV is 0.7249

.

2. Where salmonids in the Genus Oncorhynchus are present, EPA ’s recommended acute

criteri on magnitude is protect ive of the commercially and recreationally im portant adult rainbow

trout , which becomes the most sensitive endpoint at lower temperatures (see footnotes pertaining

to the 1999 FAV in Table 7 and Appendix A)

. Vertebrate sensitivity to ammonia is independent

of temperature, while invertebrate sensitiv ity to ammonia decreases as temperature decreases.

Therefore, across all temperatures the CMC equals the lower of: a) 0. 7249 times the temperature

adjusted lowest invertebrate GMAV ( for Ellipse 23.12 mg TAN/L times 0.7249

, or 1 6. 7 6 mg

TAN/L at pH 7.0 and 20oC)

, or (b) the F AV protective of adult rainbow trout (48.21 mg TAN/L )

divided by two

, or 24.10 mg TAN/L at pH 7.0 and across all temperatures , according to the

following temperature relationship :

( ) ( ( ( ( ) 

## ) ) )

Thus, the CMC increases with decreasing temperature as a result of increased invertebrate

insensitivity until it reaches a plateau of 24.10 mg TAN/L at 15.7 °C and below , where the most

sensitive taxa is the temperature invariant rainbow trout ( Table 5a ; see also Oncor h yn c hus

present line in Figure 5a ).

## 42

3

. Where Oncorhynchus sp ecies are absent, EPA ret ains all tested species in the o rder

Salmoniformes as tested surrogate species representing untested freshwater fish res ident in the

U.S. from another o rder , but does not lower the criterion to protect them as commercially and

recreationally important species

. The lowest GMAV for a freshwater fish is 51.93 mg TAN/L

for mountain whitefish ( Prosopium williamsoni) (Table 3). Therefore, in this case, the CMC

equals the lower of: a) 0.7249 times the temperature adjusted lowest invertebrate GMAV (for

Ellipse 23.12 mg TAN/L times 0.7249

, or 16.76 mg TAN/L at pH 7.0 and 20oC) , or (b) 0.7249

times the lowest freshwater fish GMAV ( 51.93 mg TAN/L at pH 7.0 and all t emperatures ),

according to the following temperature relationship :

( ) ( ( ) 

## )

Thus, the CMC increases with decreasing temperature as a result of increased invertebrate

insensitivity until it reaches a plateau of 37.65 mg TAN/L at 10 .2 °C and below (51.93 mg

TAN/L x 0.7249), where the most sensitive taxa switches to the temperature invariant fish gen us

Prosopium ( Table 5b; see also Oncor h yn c hus absent line in Figure 5a ). Note: while the

mountain white fis h ( Prosopium williamsoni ) is a species in the same family as Oncorhynchus

sp ecies (i.e., Family: Salmonidae), it is also an appropriately sensitive surrogate species amongst

all freshwater fish in the Class Actinopterygii.

The CMC

, where Oncorhynchus sp e cies are absent

, extrapolated across both temperature and pH

is as follows:

( ( ) 

## )

4

. When a threatened or endangered speci e s occurs at a site and sufficient data indicate that it is

sensitive at 1

- hour average concentrations below the CMC, it is appropriate to consider deriving

a site

- specific criterion magnitude

. It should be noted that the dataset used to derive the 2013

## 43

am monia criteria magnitudes included some threatened or endangered species, none of which

were the most sensitive of the species tested.

In summary, at pH 7 and 20 °C the CMC is 17 mg TAN/L, as primarily determined by the

sensitivity of invertebrates. As te mperature decreases to 15.7 °C and below , invertebrates no

longer are the most sensitive taxa, and thus in this range the CMC is 24 mg TAN/L

. W here

recreationally and/or commercially important Oncorhynchus species are not present , t he CMC is

determined acco rding to statement three above

. Below 15.7 °C

, if Oncorhynchus species are not

present the criteri on continue s to increase with decreasing temperature to 10.2 °C and below

,

where the CMC is 38 mg TAN/L.

Figure 5a. 2013 Acute Criterion Magnitudes Extrapolated Across a Temperature Gradient

at pH 7.

## 44

T able 5 a . Temperature and pH

- Dependent Values of the CMC (Acute Criterion Magnitude )

– Oncorhynchus s p p. Present

.

Temperature ( ° C)

pH 0

- 14 15 16 17 18 19 20 21 22 23 24 25 26 27 28 29 30

6.5 33 33 32 29 27 25 23 21 19 18 16 15 14 13 12 11 9.9



## 6.6 31 31 30 28 26 24 22 20 18 17 16 14 13 12 11 10 9.5



## 6.7 30 30 29 27 24 22 21 19 18 16 15 14 13 12 11 9.8 9.0



## 6.8 28 28 27 25 23 21 20 18 17 15 14 13 12 11 10 9.2 8.5



## 6.9 26 26 25 23 21 20 18 17 15 14 13 12 11 10 9.4 8.6 7.9



## 7 .0 24 24 23 21 20 18 17 15 14 13 12 11 10 9.4 8.6 8.0 7.3



## 7.1 22 22 21 20 18 17 15 14 13 12 11 10 9.3 8.5 7.9 7.2 6.7



## 7.2 20 20 19 18 16 15 14 13 12 11 9.8 9.1 8.3 7.7 7.1 6.5 6.0



## 7.3 18 18 17 16 14 13 12 11 10 9.5 8.7 8.0 7.4 6.8 6.3 5.8 5.3



## 7.4 15 15 15 14 13 12 11 9.8 9.0 8.3 7.7 7.0 6.5 6.0 5.5 5.1 4.7



## 7.5 13 13 13 12 11 10 9.2 8.5 7.8 7.2 6.6 6.1 5.6 5.2 4.8 4.4 4.0



## 7.6 11 11 11 10 9.3 8.6 7.9 7.3 6.7 6.2 5.7 5.2 4.8 4.4 4.1 3.8 3.5



## 7.7 9.6 9.6 9.3 8.6 7.9 7.3 6.7 6.2 5.7 5.2 4.8 4.4 4.1 3.8 3.5 3.2 3.0



## 7.8 8.1 8.1 7.9 7.2 6.7 6.1 5.6 5.2 4.8 4.4 4.0 3.7 3.4 3.2 2.9 2.7 2.5



## 7.9 6.8 6.8 6.6 6.0 5.6 5.1 4.7 4.3 4.0 3.7 3.4 3.1 2.9 2.6 2.4 2.2 2.1



## 8 .0 5.6 5.6 5.4 5.0 4.6 4.2 3.9 3.6 3.3 3.0 2.8 2.6 2.4 2.2 2.0 1.9 1.7



## 8.1 4.6 4.6 4.5 4.1 3.8 3.5 3.2 3.0 2. 7 2.5 2.3 2.1 2.0 1.8 1.7 1.5 1.4



## 8.2 3.8 3.8 3.7 3.5 3.1 2.9 2.7 2.4 2.3 2.1 1.9 1.8 1.6 1.5 1.4 1.3 1.2



## 8.3 3.1 3.1 3.1 2.8 2.6 2.4 2.2 2.0 1.9 1.7 1.6 1.4 1.3 1.2 1.1 1.0 0.96



## 8.4 2.6 2.6 2.5 2.3 2.1 2.0 1.8 1.7 1.5 1.4 1.3 1.2 1.1 1.0 0.93 0.86 0.79



## 8.5 2.1 2.1 2.1 1.9 1.8 1.6 1.5 1.4 1.3 1.2 1.1 0.98 0.90 0.83 0.77 0.71 0.65



## 8.6 1.8 1.8 1.7 1.6 1.5 1.3 1.2 1.1 1.0 0.96 0.88 0.81 0.75 0.69 0.63 0.59 0.54



## 8.7 1.5 1.5 1.4 1.3 1.2 1.1 1.0 0.94 0.87 0.80 0.74 0.68 0.62 0.57 0.53 0.49 0.45



## 8.8 1.2 1.2 1.2 1.1 1.0 0.93 0.86 0.79 0.73 0.67 0.62 0.57 0.52 0.48 0.44 0.41 0.37



## 8.9 1.0 1.0 1.0 0.93 0.85 0.79 0.72 0.67 0.61 0.56 0.52 0.48 0.44 0.40 0.37 0.34 0.32



## 9 .0 0.88 0.88 0.86 0.79 0.73 0.67 0.62 0.57 0.52 0.48 0.44 0.41 0.37 0.34 0.32 0.29 0.27

## 45

T able 5b . Temperature and pH

- Dependent Values of the CMC (Acute Criterion Magnitude )

– Oncorhynchus s p p. Absent

.

Temperature ( ° C)

pH 0

- 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25 26 27 28 29 30



### 6.5 51 48 44 41 37 34 32 29 27 25 23 21 19 18 16 15 14 13 12 11 9.9



### 6.6 49 46 42 39 36 33 30 28 26 24 22 20 18 17 16 14 13 12 11 10 9.5



### 6.7 46 44 40 37 34 31 29 27 24 22 21 19 18 16 15 14 13 12 11 9.8 9.0



### 6.8 44 41 38 35 32 30 27 25 23 21 20 18 17 15 14 13 12 11 10 9.2 8.5



### 6.9 41 38 35 32 30 28 25 23 21 20 18 17 15 14 13 12 11 10 9.4 8.6 7.9



### 7.0 38 35 33 30 28 25 23 21 20 18 17 15 14 13 12 11 10 9.4 8.6 7.9 7.3



### 7.1 34 32 30 27 25 23 21 20 18 17 15 14 13 12 11 10 9.3 8.5 7.9 7.2 6.7



### 7.2 31 29 27 25 23 21 19 18 16 15 14 13 12 11 9.8 9.1 8.3 7.7 7.1 6.5 6.0



### 7.3 27 26 24 22 20 18 17 16 14 13 12 11 10 9.5 8.7 8.0 7.4 6.8 6.3 5.8 5.3



### 7.4 24 22 21 19 18 16 15 14 13 12 11 9.8 9.0 8.3 7.7 7.0 6.5 6.0 5.5 5.1 4.7



### 7.5 21 19 18 17 15 14 13 12 11 10 9.2 8.5 7.8 7.2 6.6 6.1 5.6 5.2 4.8 4.4 4.0



### 7.6 18 17 15 14 13 12 11 10 9.3 8.6 7.9 7.3 6.7 6.2 5.7 5.2 4.8 4.4 4.1 3.8 3.5



### 7.7 15 14 13 12 11 10 9.3 8.6 7.9 7.3 6.7 6.2 5.7 5.2 4.8 4.4 4.1 3.8 3.5 3.2 2.9



### 7.8 13 12 11 10 9.3 8.5 7.9 7.2 6.7 6.1 5.6 5.2 4.8 4.4 4.0 3.7 3.4 3.2 2.9 2.7 2.5



### 7.9 11 9.9 9.1 8.4 7.7 7.1 6.6 3.0 5.6 5.1 4.7 4.3 4.0 3.7 3.4 3.1 2.9 2.6 2.4 2.2 2.1



### 8.0 8.8 8.2 7.6 7.0 6.4 5.9 5.4 5.0 4.6 4.2 3.9 3.6 3.3 3.0 2.8 2.6 2.4 2.2 2.0 1.9 1.7



### 8.1 7.2 6.8 6.3 5.8 5.3 4.9 4.5 4.1 3.8 3.5 3.2 3.0 2.7 2.5 2.3 2.1 2.0 1.8 1.7 1.5 1.4



### 8.2 6.0 5.6 5.2 4.8 4.4 4.0 3.7 3.4 3.1 2.9 2.7 2.4 2.3 2.1 1.9 1.8 1.6 1.5 1.4 1.3 1.2



### 8.3 4.9 4.6 4.3 3.9 3.6 3.3 3.1 2.8 2.6 2.4 2.2 2.0 1.9 1.7 1.6 1.4 1.3 1.2 1.1 1.0 0.96



### 8.4 4.1 3.8 3.5 3.2 3.0 2.7 2.5 2.3 2.1 2.0 1.8 1.7 1.5 1.4 1.3 1.2 1.1 1.0 0.93 0.86 0.79



### 8.5 3.3 3.1 2.9 2.7 2.4 2.3 2.1 1.9 1.8 1.6 1.5 1.4 1.3 1.2 1.1 0.98 0.90 0.83 0.77 0.71 0.65



### 8.6 2.8 2.6 2.4 2.2 2.0 1.9 1.7 1.6 1.5 1.3 1.2 1.1 1.0 0.96 0.88 0.81 0.75 0.69 0.63 0.58 0.54



### 8.7 2.3 2.2 2.0 1.8 1.7 1.6 1.4 1.3 1.2 1.1 1.0 0.94 0.87 0.80 0.74 0.68 0.62 0.57 0.53 0.49 0.45



### 8.8 1.9 1.8 1.7 1.5 1.4 1.3 1.2 1.1 1.0 0.93 0.86 0.79 0.73 0.67 0.62 0.57 0.52 0.48 0.44 0.41 0.37



### 8.9 1.6 1.5 1.4 1.3 1.2 1.1 1.0 0.93 0.85 0.79 0.72 0.67 0.61 0.56 0.52 0.48 0.44 0.40 0.37 0.34 0.32



### 9.0 1.4 1.3 1.2 1.1 1.0 0.93 0.86 0.79 0.73 0.67 0.62 0.57 0.52 0.48 0.44 0.41 0.37 0.34 0.32 0.29 0.27

## 46

Chronic c riterion c alculations

The thirty

- day rolling average concentration of total ammonia nitrogen (in mg TAN/L ) is

not to exceed, more than once every three years on the average, the chronic criterion magnitude

(CCC ) calculated using the following equation:

( ) ( ( ( ) ) 

## )

In addition, the highest four

- day average within the 30

- day averaging period should not

be more than 2.5 times the CCC (e.g., 2.5 x 1.9 mg TAN/L at pH 7 and 2 0 ° C or 4.8 mg TAN/L )

more than once in three years on ave rage

.

The 2013 CCC equation is predicated on the following:

1. The lowest GMC V in this criteria update is for an invertebrate species; thus, the CC C is both

pH

- and temperature

- dependent (based on the invertebrate chronic temperature relationship )

.

The lowest GMCV is 2.126 mg TAN/L for Villosa iris ( Table 4 ). The updated C CC ( rounded to

4 significant figures) of 1.887 mg TAN/L at pH 7and 20 ° C is 11.2 percent lower than the lowest

GMCV. The CCC to lowest GMCV ratio is 0. 8876

.

2. The most sens itive freshwater fish to chronic ammonia exposure are early life stages of

Lepomis with a GMCV of 6.920 mg TAN/L (Table 4 ). At a pH of 7 and temperature of 7 ° C and

below, the CCC plateaus (see Figure 5b) at 4.363 mg TAN/L , which is lower than the GMCV for

Lepomis

, the most sensitive fish

, multiplied by the CCC to lowest GMCV ratio ( or 6.920 mg

TAN/L x 0.8876 = 6.142 mg TAN/L); thus, a t pH 7 , the CCC is expressed as:

( ( ( ) ) 

## )

This function increases steadily with decreasing temperature (T), until it reaches a maximum at

7°C, below which it remains constant ( see Table 6; also shown graphically in Figure 5b ). The

rationale for the 7°C plateau in extrapolated invertebrate sensitivities is described in detail in

## 47

Appen dix M

. T he assumption of invertebrate insensitivity to temperatures of 7°C and below is

based on an interpretation of the empirical relationship between acute ammonia toxicity of

invertebrates and temperature, first described by Arthur et al. (1987), and in Appendix M ).

3

. All new chronic fish data added to this update of the freshwater AWQC for ammonia are

from early life

- stage tests of the species (see new data for Oncorhynchus clark i i h enshawi

,

Oncorhynchus mykiss

, Esox lucius , and Cyprinus carpio in Appendix B ), and since the new

chronic criterion magnitude lies far below all chronic values for these fishes (as well as for

Lepomis sp p. ) , the early life stage of fish no longer warrants special consideration.

4

. Where a threatened or endangered species occurs at a site and sufficient data indicate that it is

sensitive at concentrations below the CCC, it is appropriate to consider deriving a site

- specific

criterion magnitude

.

In summary, at pH 7 and 20 °C the CCC of 1.9 mg TAN/L is determined by the

sensitivity of invertebrates. As temperature decreases, invertebrate sensitiv ity to ammonia

decreases until the CCC reaches a maximum of 4.4 mg TAN/L a t pH 7 and temperature of 7 ° C

and below

.

## 48

Figure 5b. 2013 Chronic Criterion Magnitude s Extr a polated Across a Temperature

Gradient at pH 7.

## 49

Table 6 . Temperature and pH

- Dependent Values of the CCC (Chronic Criterion Magnitude ).

Temperature ( ° C)

pH 0

- 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25 26 27 28 29 30

6.5 4.9 4.6 4.3 4.1 3.8 3.6 3.3 3.1 2.9 2.8 2.6 2.4 2.3 2.1 2.0 1.9 1.8 1.6 1.5 1.5 1.4 1.3 1.2 1.1



### 6.6 4.8 4.5 4.3 4.0 3.8 3.5 3.3 3.1 2.9 2.7 2.5 2.4 2.2 2.1 2.0 1.8 1.7 1.6 1.5 1.4 1.3 1.3 1.2 1.1



### 6.7 4.8 4.5 4.2 3.9 3.7 3.5 3.2 3.0 2.8 2.7 2.5 2.3 2.2 2.1 1.9 1.8 1.7 1.6 1.5 1.4 1.3 1.2 1.2 1.1



### 6.8 4.6 4.4 4.1 3.8 3.6 3.4 3.2 3.0 2.8 2.6 2.4 2.3 2.1 2.0 1.9 1.8 1.7 1.6 1.5 1.4 1.3 1.2 1.1 1.1



### 6.9 4.5 4.2 4.0 3.7 3.5 3.3 3.1 2.9 2.7 2.5 2.4 2.2 2.1 2.0 1.8 1.7 1.6 1.5 1.4 1.3 1.2 1.2 1.1 1.0



### 7 .0 4.4 4.1 3.8 3.6 3.4 3.2 3.0 2.8 2.6 2.4 2.3 2.2 2.0 1.9 1.8 1.7 1.6 1.5 1.4 1.3 1.2 1.1 1.1 0.99



### 7.1 4.2 3.9 3.7 3.5 3.2 3.0 2.8 2.7 2.5 2.3 2.2 2.1 1.9 1.8 1.7 1.6 1.5 1.4 1.3 1.2 1.2 1.1 1.0 0.95



### 7.2 4.0 3.7 3.5 3.3 3.1 2.9 2.7 2.5 2.4 2.2 2.1 2.0 1.8 1.7 1.6 1.5 1.4 1.3 1.3 1.2 1.1 1.0 0.96 0.90



### 7.3 3.8 3.5 3.3 3.1 2.9 2.7 2.6 2.4 2.2 2.1 2.0 1.8 1.7 1.6 1.5 1.4 1.3 1.3 1.2 1.1 1.0 0.97 0.91 0.85



### 7.4 3.5 3.3 3.1 2.9 2.7 2.5 2.4 2.2 2.1 2.0 1.8 1.7 1.6 1.5 1.4 1.3 1.3 1.2 1.1 1.0 0.96 0.90 0.85 0.79



### 7.5 3.2 3.0 2.8 2.7 2.5 2.3 2.2 2.1 1.9 1.8 1.7 1.6 1.5 1.4 1.3 1.2 1.2 1.1 1.0 0.95 0.89 0.83 0.78 0.73



### 7.6 2.9 2.8 2.6 2.4 2.3 2.1 2.0 1.9 1.8 1.6 1.5 1.4 1.4 1.3 1.2 1.1 1.1 0.98 0.92 0.86 0.81 0.76 0.71 0.67



### 7.7 2.6 2.4 2.3 2.2 2.0 1.9 1.8 1.7 1.6 1.5 1.4 1.3 1.2 1.1 1.1 1.0 0.94 0.88 0.83 0.78 0.73 0.68 0.64 0.60



### 7.8 2.3 2.2 2.1 1.9 1.8 1.7 1.6 1.5 1.4 1.3 1.2 1.2 1.1 1.0 0.95 0.89 0.84 0.79 0.74 0.69 0.65 0.61 0.57 0.53



### 7.9 2.1 1.9 1.8 1.7 1.6 1.5 1.4 1.3 1.2 1.2 1.1 1.0 0.95 0.89 0.84 0.79 0.74 0.69 0.65 0.61 0.57 0.53 0.50 0.47



### 8 .0 1.8 1.7 1.6 1.5 1.4 1.3 1.2 1.1 1.1 1.0 0.94 0.88 0.83 0.78 0.73 0.68 0.64 0.60 0.56 0.53 0.50 0.44 0.44 0.41



### 8.1 1.5 1.5 1.4 1.3 1.2 1.1 1.1 0.99 0.92 0.87 0.81 0.76 0.71 0.67 0.63 0.59 0.55 0.52 0.49 0.46 0.43 0.40 0.38 0.35



### 8.2 1.3 1.2 1.2 1.1 1.0 0.96 0.90 0.84 0.79 0.74 0.70 0.65 0.61 0.57 0.54 0.50 0.47 0.44 0.42 0.39 0.37 0.34 0.32 0.30



### 8.3 1.1 1.1 0.99 0.93 0.87 0.82 0.7 6 0.72 0.67 0.63 0.59 0.55 0.52 0.49 0.46 0.43 0.40 0.38 0.35 0.33 0.31 0.29 0.27 0.26



### 8.4 0.95 0.89 0.84 0.79 0.74 0.69 0.65 0.61 0.57 0.53 0.50 0.47 0.44 0.41 0.39 0.36 0.34 0.32 0.30 0.28 0.26 0.25 0.23 0.22



### 8.5 0.80 0.75 0.71 0.67 0.62 0.58 0.55 0.51 0.48 0.45 0.42 0.40 0.37 0.35 0.33 0.31 0.29 0.27 0.25 0.24 0.22 0.21 0.20 0.18



### 8.6 0.68 0.64 0.60 0.56 0.53 0.49 0.46 0.43 0.41 0.38 0.36 0.33 0.31 0.29 0.28 0.26 0.24 0.23 0.21 0.20 0.19 0.18 0.16 0.15



### 8.7 0.57 0.54 0.51 0.47 0.44 0.42 0.39 0.37 0.34 0.32 0.30 0.28 0.27 0.25 0.23 0.22 0.21 0.19 0.18 0.17 0.16 0.15 0.14 0.13



### 8.8 0.49 0.46 0.43 0.40 0.38 0.35 0.33 0.31 0.29 0.27 0.26 0.24 0.23 0.21 0.20 0.19 0.17 0.16 0.15 0.14 0.13 0.13 0.12 0.11



### 8.9 0.42 0.39 0.37 0.34 0.32 0.30 0.28 0.27 0.25 0.23 0.22 0.21 0.19 0.18 0.17 0.16 0.15 0.14 0.13 0.12 0.12 0.11 0.10 0.09



### 9 .0 0.36 0.34 0.32 0.30 0.28 0.26 0.24 0.23 0.21 0.20 0.19 0.18 0.17 0.16 0.15 0.14 0.13 0.12 0.11 0.11 0.10 0.09 0.09 0.08

## 50

Additional e xplanation and j ustification s upporting the 2013 t emperature and pH

d ependent c alculations and c riteria m agnitudes

P art of a criterion magnitude derivation is the estimation of the CMC or CCC based on

the set of toxicity values available for different genera. The CMC or CCC estimate is intended to

be equivalent to what would be obtained by simple inspection if many genera had been tested.

Generally, the CMC or CCC is below the lowest value. For small datasets (<19) it is assumed

that the fifth percentile is lower than the lowest toxicity value. Because the CMC is one half of

the fifth percentile (i.e., FAV /2), it is frequently lower than the lowest GMAV even in large

datasets. Because the extrapolation procedure used to calculate the FAV or FCV/ CCC is based

on the slope of the four most sensitive genera , when there are data for less than 59 genera , if the

genera vary widely in sensitivity, the extrapolated criterion value is further below the lowest

value than if the criteria were tightly grouped. This is statistically appropriate because when

variance is high (i.e., values are widely spaced), the fifth pe rcentile of the distribution would be

expected to lie further below the lowest value of a small dataset than if the variance was low.

T his extrapolation procedure, while appropriate for criteria derivations across chemicals

with different variances for gen us sensitivities, is not necessarily appropriate when the genera are

following different temperature or life stage dependencies. Sensitivities can change with

temperature or life stage, and as a result, the spread of the four lowest GMAVs or GMCVs, and

th e resulting degree of extrapolation to the fifth percentile of sensitivity, can also change. Rather

than develop separate sets of GMAVs and GMCVs for each temperature and re

- computing

iteratively the CMC or CCC from the four most sensitive GMAVs or GMCVs at each

temperature

- pH combination, the extrapolation approach described below was used.

This issue of extrapolation to different temperatures and pH values with regard to chronic

toxicity was addressed in the 1999 AWQC document for ammonia by first calcul ating the ratio

of the CCC to the lowest GMCV, and then applying that ratio to subsequent criteria calculations

for all possible pH and temperature combinations. The rationale of this approach was that it

offered a degree of extrapolation that was modest and reasonable given the relatively low

number of tested genera, and that it was a preferable approach to the alternative procedure of

calculating CCCs directly from new sets of GMCVs for each pH

- temperature combination, as

each combination could result in different degrees of extrapolation, some of which could be

more than 50 percent below the lowest GMCV.

## 51

In the 1999 AWQC document, the temperature extrapolations for the CCC determination

described above were conducted separately for adult fish, fish early life stages, and invertebrates.

This was because fish GMCVs are not affected by temperature, and because the most sensitive

fish species was an early life stage of Lepomis . As a consequence, even though the lowest

GMCV at 2 0 °C was for an invertebrate, a s temperature decreases, invertebrates, but not fish,

become less sensitive to ammonia, and below 14.6 ° C , fish genera become the most sensitive.

However, the above scenario is not applicable now because at the new recommended CCC (1.9

mg TAN/L ), invertebr ate genera are the most sensitive across the entire temperature range.

I n the 1999 AWQC document, the most sensitive GMAVs were for fish, and because the

sensitivities of fish to ammonia did not vary with temperature, no temperature extrapolation was

perfo rmed. In contrast, the lowest GMAVs in this document are for invertebrates, and as a

consequence, the temperature extrapolation procedure is similarly applied to the CMC as well as

the CCC.

For the reassessment of the pH

- TAN acute toxicity relationship, E PA has determined that

the current pH

- TAN acute toxicity relationship equation effectively represents the pH

- TAN

toxicity relationship for L. siliquoidea (as determined by Wang et al. 2008), as well as for other

invertebrates, P otamopyrgus antipodarum (sna il)

, M acrobrachium rosenbergii (freshwater

shrimp) , and H. azteca (amphipod), as tested by Hickey and Vickers (1994), Straus et al. (1991),

and Borgmann and Borgmann (1997), respectively. Also, for the reassessment of the

temperature

- TAN acute toxicity rel ationship, EPA similarly determined that the current

temperature

- TAN acute toxicity relationship equation effectively represents the temperature

TAN toxicity relationship for other invertebrates, P. antipodarum (snail)

, B ranchiura sowerbyi

(oligochaete) , a nd V

iviparus bengalensis (snail) as tested by Hickey and Vickers (1994) and

Sarkar (1997), respectively.

Protection of d ownstream w aters

EPA regulations at 40 CFR 131.10(b) provide that “[i]n designating uses of a water body

and the appropriate criteria for those uses, the state shall take into consideration the water quality

standards of downstream waters and ensure that its water quality standards provide for the

attainment and maintenance of the water quality standards of downstream waters.” In cases

where downstream waters are characterized by higher pH and/or temperature, or harbor more

## 52

sensitive species, ammonia criteria more stringen t than those required to protect in

- stream uses

may be necessary in order to ensure that water quality standards provide for the attainment and

maintenance of the water quality standards of downstream waters.

Considerations for site

- specific criteria deri vation

At water temperatures above 15.7 ° C, the 2013 acute criteria magnitude is based on

effects to freshwater unionid mussels. However, when the temperature is below 15.7 ° C, and

salmonids are present (even when mussels or other sensitive mollusk species are present)

,

salmonid sensitivity determines the acute criterion (regardless of pH), similar to the 1999 acute

criterion, which was based on effects on salmonids ( i.e

. , adult rainbow trout). Where unionid

mussels and other sensitive related mollusk speci es are absent, the commercially and

recreationally important adult rainbow trout is the most sensitive species

. Site

- specific criteria

derivation must take into account the temperature at the site

. As an example, the acute criterion

magnitude at pH 7 and temperature 20 ° C cannot exceed 24 mg TAN/L (the rainbow trout SMAV

of 48.21 mg TAN/L, divided by two , used in this 2013 update as being representative of the most

sensitive fish in genera l )

.

The 1999 chronic criterion (CCC) magnitude was based on the effects on fish early life

stages, whereas based on the new data, the 2013 CCC magnitude is based on the effects on

sensitive invertebrate genera , including unionid mussels

. When mussels are present, the 2013

CCC magnitude is protective of fish early life stages regardless of temperature. See Appendix N

for additional information on site

- specific criteria for ammonia.



# EFFECTS CHARACTERIZATION

The purpose of this section is to characterize the potential effects of ammonia on aquatic

life considering available test data and to describe additional lines of evidence not used directly

in the criteria calculations

, but which support the 2013 aquatic life criteria values. This section

will also provide a summa ry of the uncertainties and assumptions, as well as provide

explanations for decisions regarding data acceptability and usage in the effects assessment. In

addition, this section will describe substantive differences between the 1999 ammonia AWQC

and the 2013 update resulting from the incorporation of the latest scientific knowledge.

## 53

All acceptable acute and chronic values for freshwater aquatic animal species, including

those from the 1999 AWQC document (re

- normalized to pH 7 and 20 ° C in the case of

inver tebrates) , are presented in Appendices A (acute) and B (chronic). These tables include new

acute and chronic ammonia toxicity data for freshwater mussels in the Family Unionidae and

reflect the latest science informing the determinations regarding accepta ble test conditions and

associated data for glochidia and juvenile mussels.



# Freshwater Acute Toxicity Data

Acute t oxicity d ata for f reshwater m ussels and n on

- pulmonate ( g ill

- bearing) s nails

Prior to publishing the 2009 draft ammonia AWQC, c oncerns ha d been raised about the

appropriateness of using data obtained from tests conducted with the parasitic glochidia life

stage of freshwater unionid mussels

. G lochidia of different species have different life history

strategies for finding an appropriate fish host ; glochidia may be free living in the water column

(and potentially exposed to pollutants) for a duration ranging from seconds to days, depending

on the particular species. EPA concluded it was useful to consider potential adverse effects on

glochidi a, because effects of chemicals on this early life stage of mussels could potentially have

broad impacts on mussel populations. In order for the toxicity test results with glochidia to be

ecologically relevant, the duration of the acute toxicity test must be comparable to the duration of

the free

- living stage of the glochidia prior to attach ing to a host

. Supported by research

conducted by Bringolf et al. (2013) demonstrating the appropriate duration of exposure for this

life stage, acceptable acute toxic ity data f or glochidia with an exposure duration of 24 hours or

less have been included in this 201 3 AWQC Update , with the stipulation that control survival for

the time period used is at least 90%

.

In addition to the four sensitive bivalve mollusk species in Table 7, there are three other

unionid mussel species among the seven genera found to be most acutely sensitive to ammonia

as well as two non

- pulmonate snail species are ranked tenth and twelfth in sensitivity . These

GMAVs represent mollusk toxicity data that w ere not in the 1999 acute criteria dataset.

New test data for the ellipse ( Venustaconcha ellipsiformis ) , the most sensitive species

tested, was not directly used in the acute criterion calculation because the methodology used

calculated the acut e value using the second through fifth most sensitive species to approximate

effects for a 5 th 

## percentile of species as noted in the effects assessment . The GMAV for ellipse

## 54

(23.12 mg TAN/L at pH 7 and 20 ° C ) is greater than the acute criterion of 17 mg TA N /L and thus

provides additional evidence supporting the protectiveness of the calculated acute criterion

(Table 3).

A vailable data on non

- pulmonate snails show that they are ano ther taxon within the

Phylum Mollusca sensitive to ammonia in freshwater ecos ystems

. The calculated GMAV of

62.15 mg TAN/L for Fluminicola sp. (pebblesnail) is the tenth most sensitive in the acute dataset

(Table 3). Another non

- pulmonate snail species Pleurocera uncialis (pagoda hornsnail) was

ranked 12 th 

## in acute sensitivity. The LC 50 

## for P. uncialis ( reported in Goudreau et al. 1993 )

,



## normalized to pH 7 and 20 ° C , is 68.54 mg TAN/L (Appendix A). To date, few studies have

been attempted with this group of species; additional testing would improve understanding of

their relative sensitivity to ammonia compared to other aquatic animals.

The draft 2009 acute criteri on magnitude recommended for waters with mussels present

was slightly higher than the 2013 acute criteria (19 vs. 17 mg TAN/L at pH 7, T 20 ° C) due to a

number of differe nces in the data used in the CMC derivations. In response to comments

received on the 2009 draft criteria , EPA removed the six invasive species from the acute dataset

for ammonia; one of the invasive species removed from the dataset was Asiatic clam, whic h had

been ranked as the fourth most sensitive GMAVs in the 2009 draft AWQC. Because the acute

dataset for ammonia is extensive and contains toxicity data for other bivalves that are native

North American species, the Asiatic clam was not needed as a biva lve surrogate. Also in the

2013 CMC, the most sensitive GMAV used to derive the CMC is for Lasmigon a subviridis

(green floater mussel) (GMAV=23.41 mg TAN/L ) which is lower than the lowest GMAV (32.73

mg TAN/L ) in the draft 2009 CMC used in the derivation of the CMC. Based on new scientific

information regarding determination of test acceptability, EPA included acceptable data for

glochidia (mussel larvae) and Hyalella azteca , which added five GMAVs for derivation of the

CMC. Since the number of GMAVs is a factor in the equation used to derive the CMC, a change

to the number of GMAVs results in a change in the resulting FAV and CMC.

## 55

Table 7

. Comparison of the F our T axa U sed to C alculate the FAV and CMC in the 1999, 2009 Draft and 201 3 AWQC

.

1999 Update CM C Magnitude 2009 Draft Update CM C Magnitude 2013 Final CM C Magnitude

Species pH 8.0,

T=25 ° C

(mg TA N/L) 

## pH 7.0,

T=20 ° C

(mg TAN /L) 

## Species pH 8.0,

T=25 ° C

( mg TAN/L ) 

## pH 7.0,

T=20 ° C

( mg TAN/L ) 

## Species pH 7.0,

T=20 ° C

( mg TAN/L )



## Oncorhynchus sp.

(salmonids), includes:

O. aquabonita, O. clarkii,

O. gorbuscha, O. kisutch,

O. mykiss , and

O. tshawytscha 21.95 99.15 Oyster mussel

, Epioblasma capsaeformis 6.037 39.24 Lampsilis sp.

(Unionidae), includes:

L. abrupta, L. cardium,

L. fasciola, L. higginsii, L.

rafinesqueana , and

L. siliquoidea 46.63

Orangethroat darter

, Etheostoma spectabile 17.96 74.25 Asiatic clam,

Corbicula fluminea 6.018 39.12 Rainbow mussel,

Villosa iris 34.23

Golden shiner,

Notemigonus crysoleucas 14.67 63.02 Lampsilis sp.

(Unionidae), includes:

L. abrupta, L. cardium,

L. fasciola, L. higginsii,

L. rafinesqueana , and

L. siliquoidea 5.919 38.48 Oyster mussel

, Epioblasma capsaeformis 31.14

Mountain whitefish,

Prosopium williamsoni 12.11 51.93 Rainbow mussel,

Villosa iris 5.036 32.73 Green floater,

Lasmigona subviridis 23.41

FAV 1 

## 11.23 48.21 FAV 5.734 37.27 FAV 33.52

CM C 5.6 24 CM C 2.9 19 CM C 17

1 

### The FAV in the 1999 AWQC document of 11.23 mg TAN/L at pH 8 was lowered to the geometric mean of these seven LC50 values at t he time in order to

protect large rainbow trout which were shown in Thurston and Russo (1983) to be measurably more sensitive than other life stages. The FAV prior to adjusting

it to protect the commercially and recreationally important adult rainbow trout was calculated to be 14.32 mg TAN/L (CMC = 7. 2 mg TAN/L) at pH 8. This

FAV based on protection of adult rainbow trout at pH 7 is 48.21 mg TAN/L (see also Appendix A in this document ).

## 56



# Freshwater Chronic Toxicity Data

Use of 28

- day j uvenile u nionid m ussel d ata

EPA decided that g rowth data from 28

- day tests with juvenile unionid mussels presented

in the Wang et al. studies from 2007 and 2011 would not be used in calculating the 2013 chronic

criterion. The decision not to use the growth data was based on the uncertainty in the tes t

method s for assess ing the growth endpoint and the need, as stated by the authors

, for additional

research “to optimize feeding conditions, to conduct longer

- term exposures (e.g., 90 d), and to

compare growth effect to potential reproductive effect in par tial li f e

- cyc l e exposure” (Wang et al.

2011). The growth endpoint showed a high degree of variability

, and the test methods for

assess ing growth, based on substrate or water

- only exposures, are currently being evaluated . For

these reasons, the survival data for 28

- day juvenile mussels w ere used in the calculation of the

CCC and not the growth data. Appendix G provides the TRAP EC 20 

## s for survival for rainbow

mussel and both Lampsilis sp ecies

, and a comparison to the growth of fatmucket mussel from

28

- day tests reported by Wang et al. (2007a , 2011 )

, which shows the additional uncertainty in the

concentration

- response relationship based on growth.

28

- day t oxic ity d ata for f reshwater s nails

As noted in the 2009 draft ammonia criteria document, n on

- pulmonate snails have been

demonstrated to be sensitive to ammonia in freshwater ecosystems, in addition to other taxa

within the Phylum Mollusca

. Besser et al. (2010) data f rom a repeat test with pebblesnail

(referred to in this document as Besser 2011) support the conclusion that non

- pulmonate snails

may be slightly less sensitive to ammonia than freshwater mussels. Additional toxicity tests are

recommended for non

- pulmonate snails and other freshwater mollusk s to further substantiate the

findings from the 28

- d ay tests summarized in Appendi ces H and I

. The calculated EC 20 

## values

using TRAP for P. idahoensis

, F. aldrichi , and T. serptenticola , and the recommended 28

- day

ammonia survival effects concentration of < 7.667 mg TAN/L for P. canaliculata , are considered

representative of non

- pulmonate snail sensitivity in general and are included in Appendix C for

the purpose of comparison.

Based on the 28

- day toxicity test results for the gill

- bearing, non

- pulmonate snail species

( Appendices B and C), EPA concludes that t he overall sensitivity of this particular group of snail

species (Sub

- class Prosobranchia, Order Neotaeni oglossa) appears high

. Furthermore, t he

## 57

sensitivity of juvenile and adult mixed

- age non

- pulmonate snails to ammonia may be greater

than that of thei r air

- breathing, pulmonate counterparts such as L. stagnalis

.

Although the GMCV for Fluminicola sp ecies is not ranked as one of the four most

sensitive used to calculate the FCV, the value is ranked the 5 th 

## most sensitive in the chronic

criterion dataset

. T he 28

- day growth E C 20 

## for this freshwater non

- pulmonate snail species

,

calculated using EPA’s TRAP

, is 2. 281 mg TAN/L at test pH (8.22) and temperature (20.1 ° C),

or 7.828 mg TAN/L after adjustment to pH 7 and 2 0 ° C (see Appendix B ). Appendix H includes

a summary of the 28

- day toxicity test results for Fluminicola sp ecies which are acceptable for

use quantitatively in the chronic dataset. The TRAP output for this test is provided in Appendix

H

.

28

- day t oxicity d ata for Hyalella azteca : M i nimum D ata R equirement N umber 5

Literature data indicate that the response of Hyalella azteca is influenced not only by pH,

but also by sodium concentration in the dilution water. Borgmann and Borgmann (1997)

demonstrated that increasing sodium decreased the toxicity of ammonia to Hyalella , and applied

these findings to explain differences in toxicity observed by Ankley et al. (1995), which were

originally attributed to water hardness. Further unpublished experiments by EPA’s Office of

Research and Development confir m Borgmann’s assertion that sodium concentration plays a key

role in determining the acute response of Hyalella to ammonia (personal communication, D.R.

Mount , EPA, ORD ). Because sodium is not known to affect ammonia toxicity to other species,

this criter ion does not consider sodium concentration, and this variation is not explicitly

addressed. For purposes of deriving a GMAV for Hyalella , tests were selected that had a

moderate sodium concentration (e.g., “moderately hard” water tests from Ankley et al. 1995, see

Appendix A), and tests with extremely low sodium concentrations were excluded (e.g., “soft”

water tests from Ankley et al. 1995; data from Whiteman et al. 1996, see Appendix J). The

available acute data for ammonia did not include tests conducte d in natural waters with a sodium

concentration below about 3 mg/L; at that sodium concentration, the acute values for Hyalella

were near the FAV reported in this document. Whether acute toxicity of ammonia to Hyalella

would occur below the FAV in waters with less than 3 mg/L sodium is unknown ( Appendix H).

For the 2013 chronic criterion , EPA re

- evaluated the available data for Hyalella azteca

based on recent research regarding the appropriate test conditions

, including water chemistry

## 58

(e.g., appropriate concentration s for specific ions such as chloride) and feeding regimes. The

concentrations of sodium are important to H. azteca health as discussed previously and the

sodium concentrations in the chronic test used in the CCC represent approximately the m id

range of U.S. waters. Based on this re

- evaluation, EPA determined that certain tests met the new

recommended conditions that would support healthy test organisms

, and accepted those data for

use in the calculation of the CCC. The specific tests used were from Borgmann (1994); details

on these tests are included in Appendix H under Chronic Toxicity Tests with Juvenile Hyalella

azteca

. As a result of inclusion of acceptable H. azteca data

, the minimum data requirement

( MDR ) for a benthic crustacean is fulfilled for the chronic criterion provided in this document

.

The GMCV of 29.17 mg TAN/L ranks Hyalella azteca as the thirteenth (of 16) most sensitive

GMCV

.

## 59

Reconsideration of the c hronic t oxicity d ata a vailable for a quatic i nsects : Minimum D ata

R equirement N umber 6

EPA chose not to include a chronic value for the stonefly, Pteronarcella badia , from

Thurston et al. (1984a) in the 1999 AWQC update document because this aquatic insect species

is relatively insensitive . Upon further consideration, EC 20 

## s for 30

- day nymph mortality were

calculated for field collected Pteronarcella badia for two separate partial life cycle tests in

consecutive years , in order to develop a GMCV for insects to fulfill the six th minimum data

requirement (MDR) , and to clearl y specify the expected lack of sensitivity of insects to ammonia

based on available data. EC 20 

## s were calculated for each test

, and, a s the authors themselves

noted, the results were variable between the two tests. The normalized EC 20 

## for the test

conduct ed with P. badia collected from the Gallatin R. was 207.0 mg TAN/L, and was 26.27 mg

TAN/L for the test conducted with P. badia collected from the Rocky River (Appendix B). The

geometric mean for these two tests is 73.74 mg TAN/L

. I t is not known if thes e tests were

conducted using the most sensitive life stage ; however, t he authors did note that the length of

individuals u sed in both tests was similar. EPA used the two EC 20 

## s based on 30

- day nymph

mortality to calculate a GMCV of 73.74 mg TAN /L for this species. T he stonefly is listed as

sixteenth GMCV in chronic sensitivity , fulfilling the sixth MDR. Additional data on insect

sensitivity to ammonia would be useful in confirming the conclusion that insects are relative ly

insensitiv e to ammonia in freshwater environments, given the limited data available.

New c hronic t oxicity d ata for s almonid s pecies and d erivation of a GMCV for

Oncorhynchus : Minimum D ata R equirement N umber 1

Chronic values f

rom two additional studies with Oncorhynchus specie s (salmonids) are

included in this AWQC document that we re not included in the 1999 document (see Appendix H

for more detailed description s of the results fr om these studies ). Koch et al. (1980) exposed

Lahontan cutthroat trout ( Oncorhynchus clark i i henshawi ) , an endangered species, for 103 days

in an ELS test. There were no successful hatches at exposure levels of 148 mg TAN/L or higher

and no significant mortality at exposure levels below 32.9 mg TAN/L . Regression analysis of

the survival data res ulted in a calculated EC 20 

## value of 17.89 mg TAN/L at 13.7 ° C and pH 7.57

.

The EC 20 

## value is 25.83 mg TAN/L when adjusted to pH 7 ( Appendix B ).

## 60

The results of a more recent 90

- day ELS test using a wild strain of rainbow trout exposed

to ammonia were reported by Brinkman et al. (2009) , a nd are included in this criteria document.

The test was initiated with newly fertilized embryos exposed under flow

- through conditions

through hatch, swim

- up, and early fry development. Survival, growth and biomass of swim

- up

fry were significantly reduced at 16.8 mg TAN/L compared to controls, but unaffected at 7.44

mg TAN/L . The EC 20 

## calculated for biomass using TRAP and normalized to pH 7 is 15.60 mg

TAN/L (Appendix B).

I n the 1999 AWQC document, the results of six chronic tests conducted with ammonia

for Oncorhynchus mykiss and Oncorhynchus nerka were i n cluded in Table 5 of that document as

“acceptable” chronic tests for criteria development. A GMCV was not derived for

Oncorhynchus at that time

, however, because o f the degree of variability among test results, as

well as a preponderance of “greater than” or “less than” values, preventing the calculation of

definitive SMCVs within the genus. The results of these chronic tests were only used at that

time to assess t he appropriateness of the CCC.

For this 2013 document

, these six studies and the data from the two additional studies

summarized above, have been re

- evaluated and re

- considered for inclusion in deriving a new

chronic criterion for ammonia in order to con sider and include all available, reliable toxicity test

informa tion for this recreationally, commercially and ecologically important taxon. The data

w ere re

- examined with specific consideration of whether unbounded (greater or less than) values

add relevant information to determination of final SMCVs for Oncorhynchus clarkii, O. mykiss,

and O. nerka

. A decision rule was developed for evaluating chronic values (EC 20 

## s) for potential

use in deriving an SMCV for a salmon species . In developi ng the decision rule, it was noted that

“greater than” values for concentrations of low magnitude, and “less than” values for

concentrations of high magnitude do not add significant information to the analysis. That is, if a

researcher only tested very lo w concentrations and found no chronic effects or very high

concentrations and found 100% response for a chronic endpoint, those data do not significantly

enhance understanding of the toxicity of ammonia. Conversely, if a researcher only tested very

low con centrations and found significant chronic effects, indicating the test material was highly

toxic, or relatively high concentrations and found incomplete response for a chronic endpoint,

indicating low toxicity of the materials, those data do significantly enhance understanding of the

toxicity of ammonia. Thus, the decision rule was applied as follows: “ greater than ” ( > ) low CV s

## 61

and “ less than ” ( < ) high CV s were not used in the calculation of the SMCV; but “less than” ( < )

low CV s and a “greater than” ( > ) hi gh CV s were included in the SMCV.

Following this decision rule, the SMCV for O. clarkii is the normalized EC 20 

## of 25.83 mg

TAN/L from Koch et al. (1980). The SMCV for O. mykiss is 6.663 mg TAN/L at pH 7, which is

the geometric mean of the <3.246 mg TAN/L value from Calamari et al. (1977, 1981), the

<3.515 mg TAN/L value from Solbe and Shurben (1989), the >11.08 mg TAN/L value from

Thurston et al. (1984b), and the 15.60 mg TAN/L value from Brinkman et al. (2009). W ith

respect to the SMCV for O. mykiss

, bo th the Calamari et al. (1977, 1981) and the Solbe and

Shurben (1989) values are low “less than” values, indicating demonstrated toxicity at low

concentrations of ammonia, while the Thurston et al. ( 1984b) value is a relatively high “ greater

than” value, in dicating lower toxicity to O. mykiss in this test, compared to the Calamari (1977,

1981) and Solbe and Shur b en (1989) value s with respect to the SMCV for O. mykiss. Finally,

the SMCV for O. nerka is <10.09 mg TAN/L (Rankin 1979), and has been included as a

relatively low “ less than ” value, indicating relative sensitivity to the effects of ammonia in this

test. The < 48 mg TAN/L value (at pH 7) from Thurston et al. (1978) re

- calculated for O. clarkii

(see Appendix C; value represents the geometric mean of four values) and the <45.50 mg TAN/L

value from Burkhalter and Kaya (1977) for O. mykiss (retained in Appendix B) are not included

in the SMCV calculation s because they are high “less than” value s

, and do not add imp ortant

information to the analyses

. The new GMCV for Oncor h ynchus in this 2013 Update document

is 12.02 mg TAN/L, which is the geometric mean of the three SMCVs for Oncorhynchus clarkii

( 25.83 mg TAN/L) , O. mykiss (6.663 mg TAN/L) , and O. nerka (<10.09 mg TAN/L) (Appendix

B), resulting in Oncor

h ynchus as being seventh most sensitive GMCV (Table 4).

Another o rder of i nsect or a p hylum n ot a lready r epresented : Minimum D ata R equirement

N umber 8

For the MDR identified earlier as “# 8

, ” “another order of insect or a phylum not already

represented,” there are no additional chronic toxicity data for any freshwater animal that would

fulfill this MDR in the chronic dataset (the acute dataset fulfills all eight MDRs). Therefore,

EPA developed a surrogate ammonia CV for the Phylum Annelida by using the geometric mean

acute value from the four available genera ( Dendrocoelus, Limnodrilus, Lumbriculus, and

Tubifex ) and applying an ACR from other invertebrate groups

. There is less than a factor of two

## 62

difference between the GMAVs for the most ( Dendrocoelus , 119.5 mg TAN/L ) and least

( Lumbriculus, 218.7 mg TAN/L ) sensitive genus (see Table 3) . A surrogate chronic value was

derived by dividing the GMAV for all four annelids (176.2 mg TAN/L ) by a geometric mean

species leve l invertebrate acute to chronic ratio (6.320), represented by pelagic crustaceans

(cladocerans), a benthic crustacean (amphipod) and prosobranch snail (see Appendix F Acute

Chronic Ratios). The resulting surrogate CV for Phylum Annelida is 27.88 mg TAN/L (at pH 7

and 20 ° C).



# Protection of Endangered Species

The dataset for ammonia is particularly extensive and includes data representing species

that are Federally

- listed as threatened or endangered by the U.S. Fish and Wildlife Service and/or

NOAA National Marine Fisheries Service . Summaries are provided here describing the data for

the listed species and demonstrating that the 2013 ammonia criteria update is protective of these

species, based on best available scientific data.

Key a cute t oxicity d ata for l isted s pecies

The acute criterion dataset for ammonia now includes 1 2 aquatic species that are

Federally

- listed as threatened, endangered or of concern.

For unionid mussels

, the 2013 criterion acute dataset includes acceptable data for 16

freshwater sp ecies across 11 genera

. Of these

, f ive of the mussel species are Federally

- listed as

threatened or endangered (as identified in Table 3 ). The oyster mussel ( Epioblasma

capsaeformis ) is a Federally

- listed species and is the third most sensitive in the acute dataset

with a GMAV , based on a single SMAV

, of 31.14 mg TAN/L

. The SMAV/2 for the oyster

mussel is approximately 16 mg TAN/L

, similar to the 2013 acute criterion value of 17 mg

TAN/L

. The S MAV/2 is a value considered statistically indistinguishabl e from control mortality

or effect based o n analysis of 219 acute toxicity tests on a range of chemicals, as described in the

Federal Register on May 18, 1978 (43 FR 21506

- 18). Thus, the magnitude of acute effects to

this species at the SMAV/2 are not exp ected to significantly impact the species, because it is

expected to be statistically indistinguishable from effects to control (unexposed) organisms.

Furthermore, the acute criterion specifies that this concentration should not be exceeded for more

than one hour once every three years on average, providing further protection through the

## 63

limita tion of any excursions above the criterion. Thus, t he 2013 recommended CMC for

ammonia of 17 mg TAN/L should be protective of oyster mussels . In waters where this listed

species is present, a site

- specific criterion could be considered using the SMAV for that species

as the FAV from which to derive the CMC.

The Lampsilis GMAV of 46.63 mg TAN/L reflects the geometric mean of SMAVs for

six mussel species, two ( L. abru pta and L. higginsii ) of which are endangered and a third ( L.

rafinesqueana ) that is a Federal species of concern (Table 3). The SMAVs for this genus range

from 26.03 mg TAN/L ( L. abrupta ) to 69.97 mg TAN/L ( L. rafinesqueana ) (Appendix A).

Given the range of sensitivity within this genus with listed species at both the most and le ast

sensitive end s of the range, the CMC of 17 mg TAN/L should be protective of the genus as a

whole , with SMCVs/2 ranging from 13 to 34 mg TAN/L

. Again, the acute criterion specifies

that a concentration of 17 mg TAN/L should not be exceeded for more than one hour once every

three years on average, providing further protection of species, through the limitation of any

excursions above the criterion

. In waters where the listed species are present, a site

- specific

criterion could be considered using the SMAV for that species as the FAV from which to derive

the CMC.

Also among the ten most sensitive GMAVs in the acute dataset is the GMAV for the

endan gered Lost River sucker ( Deltistes luxatis ) endemic to the Klamath Basin of northern

California and southern Oregon (Appendix A). The reported LC 50 

## s at test temperature 20 ° C and

pH 8.0 were 10.35 and 16.81 mg TAN/L for larval and juvenile fish, expressed as total ammonia

(Appendix A). The LC 50 

## s normalized to pH 7 and 20 ° C are 44.42 and 72.18 mg TAN/L

,

respectively (Appendix A). The GMAV for Lost River sucker is calculated as the geometric

mean of the two normalized LC 50 

## s, or 56.62 mg TAN/L (Table 3) , wit h an SMAV/2, or expected

low mortality level, of approximately 28 mg TAN/L , significantly above the CMC

. Lost River

sucker represents the ninth most sensitive genus in the acute dataset, and second most sensitive

fish species (following mountain whitefish which is the most sensitive fish GMAV)

, and thus is

expected to be protected by the CMC of 17 mg TAN/L

.

The second most acutely sensitive salmonid genus (after Prosopium

, represented by th e

mountain whitefish, Prosopium williamsoni

, acute sensitivity rank 8) is Oncorhynchus

,

represented by data for six different species, three of which are threatened or endangered, with

SMAVs ranging from 78.92 mg TAN/L for Cutthroat trout, O. clarkii

, to 180.7 mg TAN/L for

## 64

pink salmon, O. gorbuscha

. The G MAV for Oncorhynchus (99.15 mg TAN/L ) is ranked #25 in

acute sensitivity (Table 3). All SMAV/2 values for the threatened or endangered species tested

in this genus are significantly above the acute criterion magnitude . Thus, the acute criterion is

expect ed to be protective of threatened and endangered salmonid species.

Key c hronic t oxicity d ata for l isted s pecies

In the chronic dataset for ammonia, the Federally

- listed species are represented by three

salmonid species in the genus Oncorhynchus , including sockeye salmon, rainbow trout, and the

subspecies Lahontan cutthroat trout. The GMCV for Oncorhynchus of 12.02 mg TAN/L

includes the three SMCVs ranging from 6.663 (rainbow trout) to 25.83 mg TAN/L (Lahontan

cutthroat trout) (Table 4). The CC C for ammonia of 1.9 mg TAN/L is expected to be protective

of this genus as a whole

. At pH 7, the CCC is 3.5 times lower than the chronic value for the

most sensitive tested listed salmonid species, O. mykiss

, which includes populations of rainbow

trout a nd steelhead trout.

In addition , three other studies provide useful information with which to assess the

protectiveness of the CCC for threatened and endangered fish species (data included in Appendix

C ). All three studies indicate that the chronic crite rion is expected to be protective of the

endangered or listed species tested by the researchers, as described below.

Meyer and Hansen (2002) conducted a 30

- day toxicity test with late

- stage larvae (0.059

g) of Lost River suckers ( Deltistes luxatus ) at pH 9.5. The exposure duration and pH were

chosen to represent the period of combined elevated unionized ammonia concentrations and

elevated pH that occur during cyanobacterial blooms in surface waters of Upper Klamath Lake,

which have been shown to last for several weeks to a month. Survival decreased significantly at

1.23 and 2.27 mg TAN/L , whereas the highest NOEC for all endpoints (survival, growth, body

ions, and swimming performance) was 0.64 mg TAN/L . Control survival was > 90 percent. The

calculated LOEC of 1.23 mg TAN/L at test pH and temperature when normalized to pH 7

corresponds to a value of 25.31 mg TAN/L, again, substantially higher than the 2013 chronic

criterion value ( Appendix C ).

In order to determine if whole effluent toxicity testing is protective of threatened and

endangered fish species, Dwyer et al. (2005) conducted 7

- day chronic toxicity tests with

Ceriodaphnia dubia (neonates, <24 h old) and fathead minnow larvae ( Pimephales promelas,

## 65

<24 h) in addition to the following six threatene d a nd endangered fish species: bon ytail chub

( Gila elegans ), spotfin chub ( Erimonax , formerly Cyprinella, mon a ch us ), Cape Fear shiner

( Notropis mekistocholas ), gila topminnow ( Poeciliopsis occidentalis ), Colorado pikeminnow

( Ptychocheilus lucius ), and razo rback sucker ( Xyrauchen texanus ). The age of the six threatened

and endangered fish species used during the 7

- day ammonia exposures ranged from <1 to 7 days.

The combined effect on test species survival and growth were determined as E C 25 

## values

. The

six endangered species, presented in the same order as they are listed above, have reported E C 25



## values of: 11.0, 15.8, 8.80, 24.1, 8.90 and 13.4 mg TAN/L ; or 23.24

, 33.37

, 18.59

, 50.91

, 18.80

and 28.30 mg TAN/L when adjusted to a pH of 7 .0 ( Appendix C ). These values are all

substantially higher than the 2013 chronic criterion concentration value of 1.9 mg TAN/L

.

Based on the results, the two species typically used for whole effluent toxicity testing ( C. dubia

and P. promelas) were more sensitive to amm onia and are protective of the six listed fish species

when used as surrogate test species.

Fairchild et al. (2005) conducted 28

- day toxicity tests with early life stages of Colorado

pikeminnow ( Ptychocheilus lucius ) and razorback sucker ( Xyrauchen texanu s ), and compared

the results of those tests with a test using a surrogate fish species, the fathead minnow

( Pimephales promelas ). Effect concentrations based on the survival and growth endpoints of the

fathead minnow and razorback sucker tests were not di fferent; however, growth was the more

sensitive endpoint for the Colorado pikeminnow test. The 28

- day growth LOEC for the

Colorado pikeminnow was 8.60 mg TAN/L , or 29.75 mg TAN/L at pH 7, substantially greater

than the 2013 chronic criterion. The 28

- day survival LOEC for the razorback s ucker was 13.25

mg TAN/L , or 46.58 mg TAN/L at pH 7 . Both endangered fish species exhibited similar

sensitivity to ammonia as t he fathead minnow (LOEC of 32.71 mg TAN/L at pH 7 ; see

Appendix C )

. The same can be said for t he Lost River sucker, which indicates that these

particular endangered fish species are expected to be protected by the CCC value calculated in

this 201 3 AWQC Update.



# Comparison of 1999, 2009, and 2013 Criteria Values

Table 8 provides a comparison of the four most sensitive taxa used to calculate the CCC

in this 201 3 AWQC Update document compared to the four most sensitive taxa used to calculate

the CCC in the 1999 AWQC document.

## 66

The 2013 CCC is about twice the magnitude of the draft 2009 CCC recommended f or

waters with mussels present (1.9 vs. 0.91 mg TAN/L , respectively, at pH 7, T=20 ° C) as a result

of differences in the data used in the CCC derivations. Based on a new study by Wang et al.

(2011) described in the Effects Analysis section under Summaries of Studies Used in Chronic

Criterion Determination , pg. 34, above , the lowest GMCV for the mussel genus Lampsilis

increased from 1.154 mg TAN/L in the 2009 draft AWQC to 2.216 mg TAN/L in the 2013

AWQC. As a result, compared to the four lowest GMCVs in th e 2009 draft CCC, the four

lowest GMCVs in the 2013 CCC have a smaller range of variation in values (2.216 to 7.547 mg

TAN/L ) which decreases the uncertainty of the 5 th 

## percentile GMCV estimation. Also in the

2009 draft CCC, there were only 13 GMCVs in th e datas et used to derive the CCC while in the

2013 CCC

, there are 16 GMCVs used to derive the CCC, because of the addition of the GMCVs

for Hyalella azteca

, the insect Pteronarcella badia , and salmonids ( Oncorhynchus s p p. ). The

new GMCVs a ffect the chroni c species sensitivity distribution. The cumulative probability (P)

decreases as a function of the increased number of GMCVs and result s in an increase in the

FCV.

## 67

Table 8

. Comparison of the F our T axa U sed to C alculate the FCV and CCC in th e 1999 Update, 2009 Draft and the 201 3

AWQC

.

1999 Update CCC Magnitude 2009 Draft Update CCC Magnitude 2013 Final CCC Magnitude

Species pH 8.0,

T=25 ° C

( mg

TAN/L ) pH 7.0,

T=20 ° C

( mg

TAN/L ) Species pH 8.0,

T=25 ° C

( mg

TAN/L ) pH 7.0,

T=20 ° C

( mg

TAN/L ) Species pH 7.0,

T=20 ° C

( mg

TAN/L )

Fathead minnow,

Pimephales promelas 3.09 7.503 Long fingernail clam,

Musculium transversum <2.260 7.552 Long fingernail clam,

Musculium transversum 7.547

Lepomis sp.

(Centrarchidae), includes:

Bluegill sunfish,

L. macrochirus , and

Green sunfish,

L. cyanellus 2.85 6.92 Lepomis sp.

(Centrarchidae), includes:

Bluegill sunfish,

L. macrochirus , and

Green sunfish,

L. cyanellus 2.852 6.924 Lepomis sp.

(Centrarchidae), includes:

Bluegill sunfish,

L. macrochirus , and

Green sunfish

, L. cyanellus 6.92

Long fingernail clam,

Musculium transversum <2.26 7.547 Rainbow mussel,

Villosa iris <0.9805 3.286 Rainbow mussel,

Villosa iris 3.501

Amphipod,

Hyalella azteca <1.45 4.865 Lampsilis sp.

(Unionidae), includes:

Wavy

- rayed lamp mussel,

L. fasciola and

Fatmucket, L. siliquoidea <0.3443 1.154 Lampsilis sp.

(Unionidae), includes:

Wavy

- rayed lamp mussel,

L. fasciola and

F atmucket, L. siliquoidea 2.216

CCC 1.2 4.5 CCC 0.26 0.91 CCC 1.9

## 68

Comparison of s tatistical a pproaches to d evelop the c hronic c riterion : EC20 vs. MATC

In th is 2013 ammonia criteria update, the CCC is based on a 20 percent reduction in

survival, growth, or reproduction, which is a risk management decision made by EPA in 1999

and also retained for this document . When an EC 20 

## was not provided in a study, the EPA’s

TRAP program was used to estimate the EC 20 

## as the basis for the GMCV and included the

resultant CCC derivation of 1.9 mg TAN/L

. An alternative chronic measu re of effect that is

commonly used is the MATC, which is the geometric mean of the NOEC and LOEC . In the case

of the current ammonia dataset, using MATCs to derive the chronic criteria would result in an

FCV of 1.972 and CCC of 2.0 mg TAN/L . This compari son demonstrates that

, for the current

ammonia chronic dataset

, the use of TRAP to estimate EC 20 

## values does not result in a

significant difference from the MATC, an other statistical approach frequently used to develop

chronic effects assessments and crite ria.

T he concentrations of TAN affecting freshwater animals in this 201 3 AWQC update are

normalized to pH 7.0 for all aquatic organisms and 20 ° C for invertebrates

. I n contrast

, the

concentrations of TAN affecting freshwater animals in the 1999 AWQC were normalized to pH

8.0 for all organisms and temperature 25 ° C for invertebrates

. Th e current pH (7) and

temperature (20 ° C) used are considered to more closely reflect ambient pH and temperatures

found generally in surface waters in the U.S. The acute and chronic criteri on value s can be

adjusted to reflect local pH and temperature using the values in Tables 5a, 5b, and 6 derived

from the equations presented in The National Criteria for Ammonia in Fresh Water section

(pages 4 0

- 4 9 ).



# UNUSED DATA

For this 201 3 criteria update document, EPA considered and evaluated all available data

that could possibly be used to derive the new acute and chronic criteria for ammonia in fresh

water. A substantial amount of th ose data w ere associated with studies that did not meet the

basic QA/QC requirements described in the 1985 Guidelines (see Stephan et al. 1985) . In such

cases, EPA further scrutinized those studies where either: (1) the study included tests with a

species associated with one of the four most sensi tive GMAVs or GMCVs used to derive the

2013 criterion; or (2) the study provided results of tests where the normalized acute or chronic

## 69

value for the test was within a factor of approximately two of the fourth ranked most sensitive

GMAV or GMCV , and thus m ight be considered potentially influential to the acute or chronic

criterion . For each study that was potentially influential, but did not meet the additional data

quality requ i r e ments for its use in deriving criteria for ammonia , the study and its result s w ere

included in Appendix J (acute studies) and K (chronic studies)

, and a rationale i s provided for its

exclusion. A list of all other studies considered but removed from consideration f or use in

deriving the criteria is provided in Appendix L with a code (and in some cases comments)

indicating the reason(s) for exclusion.

## 70



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Abeliovich, A. and Y. Azov. 1976. Toxicity of ammonia to algae in sewage oxidation ponds.

Appl. Environ. Microbiol. 31(6): 801

- 806.

Abell, R.A., D.M. Olson, E. Dinerstein, P.T. Hurley, J. Diggs, W. Eichbaum, S. Walters, W.

Wettengel, T. Allnutt, C. Loucks and P. Hedao. 2000. Freshwater ecoregions of North America.

A conservation assessment. Island Press (WWF), Washington, DC.

Adams, N. and D. Bealing. 1994. Orga nic pollution: Biochemical oxygen demand and ammonia.

In: Handbook of ecotoxicology, vol. 2. Calow, P. (Ed.). Blackwell Scientific Publications,

Oxford. pp. 264

- 285.

Adelman, I.R., L.I. Kusilek, J. Koehle and J. Hess. 2009. Acute and chronic toxicity of a mmonia,

nitrite and nitrate to the endangered Topeka shiner ( Notropis topeka ) and fathead minnows

( Pimephales promelas ). Environ. Toxicol. Chem. 28(10): 2216

- 2223.

Agency for Toxic Substances and Disease Registry. 2004. A toxicological profile for ammonia.

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## 86

Appendix A. Acute Toxicity of Ammonia to Aquatic A nimals.

Appendix A. Acute Toxicity of Ammonia to Aquatic Animals

Species Chemical Name Duration Methods a pH Temp.

( ̊C) Total

Ammonia

(mg

TA N/L) Total Ammonia b

(mg TA N/L)

a djusted to pH 7

(all organisms )

and 20 ° C

( invertebrates ) SMAV

(mg

TA N/L) GMAV

(mg

TA N/L) Reference

Insect (8th

- 10th instar),

Erythromma najas Ammonium

chloride 4 d R,U 7.5 25 589 1618 Beketov 2002

Insect (8th

- 10th instar),

Erythromma najas Ammonium

chloride 4 d R,U 8.7 25 168 4163 Beketov 2002

Insect (8th

- 10th instar),

Erythromma najas Ammonium

chloride 4 d R,U 9.1 25 49.2 2361 2515 2515 Beketov 2002

Caddisfly,

Philarctus quaeris Ammonium

chloride 4 d F,M 7.8 21.9 296.5 1032 Arthur et al. 1987

Caddisfly,

Philarctus quaeris Ammonium

chloride 4 d F,M 7.8 13.3 561.7 958.4 994.5 994.5 West 1985; Arthur et al.

1987

Beetle,

Stenelmis sexlineata Ammonium

chloride 4 d F,M 8.7 25 29.7 735.9 735.9 735.9 Hazel et al. 1979

Crayfish,

Orconectes immunis Ammonium

chloride 4 d F,M 7.9 17.1 488.1 1367 Arthur et al. 1987

Crayfish (adult),

Orconectes immunis Ammonium

chloride 4 d F,M 8.2 4.6 999.4 1757 1550 West 1985; Arthur et al.

1987

Crayfish (2.78 cm),

Orconectes nais Ammonium

chloride 4 d F,M 8.3 26.5 23.15 303.8 303.8 686.2 Evans 1979

Midge (10 d old, 2

- 3 instar),

Chironomus riparius Ammonium

chloride 4 d R,M 7.7 21.7 357.7 1029 1029 Monda et al. 1995

Midge,

Chironomus tentans Ammonium

chloride 4 d S,M 6.69 23 430 443.0 Besser et al. 1998

Midge,

Chironomus tentans Ammonium

chloride 4 d S,M 7.56 23 564 1439 Besser et al. 1998

Midge (2nd instar),

Chironomus tentans Ammonium

chloride 4 d F,M 6.5 25 371 415.1 Schubauer

- Berigan et al.

1995

## 87

Appendix A. Acute Toxicity of Ammonia to Aquatic Animals

Species Chemical Name Duration Methods a pH Temp.

( ̊C) Total

Ammonia

(mg

TA N/L) Total Ammonia b

(mg TA N/L)

a djusted to pH 7

(all organisms )

and 20 ° C

( invertebrates ) SMAV

(mg

TA N/L) GMAV

(mg

TA N/L) Reference

Midge (2nd instar),

Chironomus tentans Ammonium

chloride 4 d F,M 8.1 25 78.1 614.0 Schubauer

- Berigan et al.

1995

Midge (2nd instar),

Chironomus tentans Ammonium

chloride 4 d F,M 6.5 25 368 411.7 Schubauer

- Berigan et al.

1995

Midge (2nd instar),

Chironomus tentans Ammonium

chloride 4 d F,M 8.1 25 50.5 397.0 451.8 681.8 Schubauer

- Berigan et al.

1995

Mayfly (middle to late instar),

Drunella grandis Ammonium

chloride 4 d F,M 7.84 12.8 259.1 455.5 Thurston et al. 1984b

Mayfly (middle to late instar),

Drunella grandis Ammonium

chloride 4 d F,M 7.84 13.2 195.6 355.6 Thurston et al. 1984b

Mayfly (middle to late instar),

Drunella grandis Ammonium

chloride 4 d F,M 7.85 12 319 534.5 442.4 442.4 Thurston et al. 1984b

Aquatic sowbug,

Caecidotea racovitzai

(previously Asellus racovitzai ) Ammonium

chloride 4 d F,M 7.8 22 148.8 522.3 Arthur et al. 1987

Aquatic sowbug (adult),

Caecidotea racovitzai Ammonium

chloride 4 d F,M 8 4 357.8 407.7 West 1985; Arthur et al.

1987

Aquatic sowbug,

Caecidotea racovitzai Ammonium

chloride 4 d F,M 7.81 11.9 176 272.2 387.0 387.0 Thurston et al. 1983

Isopod (adult),

Asellus aquaticus Ammonium

chloride 4 d F,M 7.05 12 2.60 575.2 Dehedin et al. 2012

Isopod (adult),

Asellus aquaticus Ammonium

chloride 4 d F,M 7.05 12 1.25 276.6 Dehedin et al. 2012

Isopod (adult),

Asellus aquaticus Ammonium

chloride 4 d F,M 7.05 12 1.70 376.1 Dehedin et al. 2012

Isopod (adult),

Asellus aquaticus Ammonium

chloride 4 d F,M 7.05 18 2.61 603.8 Dehedin et al. 2012

Isopod (adult),

Asellus aquaticus Ammonium

chloride 4 d F,M 7.05 18 1.40 323.9 Dehedin et al. 2012

Isopod (adult),

Asellus aquaticus Ammonium

chloride 4 d F,M 7.05 18 1.95 451.1 Dehedin et al. 2012

Isopod (adult),

Asellus aquaticus Ammonium

chloride 4 d F,M 7.05 24 1.00 246.6 Dehedin et al. 2012

## 88

Appendix A. Acute Toxicity of Ammonia to Aquatic Animals

Species Chemical Name Duration Methods a pH Temp.

( ̊C) Total

Ammonia

(mg

TA N/L) Total Ammonia b

(mg TA N/L)

a djusted to pH 7

(all organisms )

and 20 ° C

( invertebrates ) SMAV

(mg

TA N/L) GMAV

(mg

TA N/L) Reference

Isopod (adult),

Asellus aquaticus Ammonium

chloride 4 d F,M 7.05 24 1.00 246.6 Dehedin et al. 2012

Isopod (adult),

Asellus aquaticus Ammonium

chloride 4 d F,M 7.05 24 2.00 493.1 378.2 378.2 Dehedin et al. 2012

Threespine stickleback

(juvenile

- adult, 32

- 60 mm),

Gasterosteus aculeatus Ammonium

chloride 4 d S,M 7.1 23.3 198.1 216.5 Hazel et al. 1971

Threespine stickleback

(juvenile

- adult, 32

- 60 mm),

Gasterosteus aculeatus Ammonium

chloride 4 d S,M 7.15 15 577 667.4 Hazel et al. 1971

Threespine stickleback

(juvenile

- adult, 32

- 60 mm),

Gasterosteus aculeatus Ammonium

chloride 4 d S,M 7.25 23.3 203.8 264.0 Hazel et al. 1971

Threespine stickleback

(juvenile

- adult, 32

- 60 mm),

Gasterosteus aculeatus Ammonium

chloride 4 d S,M 7.5 15 143.9 261.1 Hazel et al. 1971

Threespine stickleback

(juvenile

- adult, 32

- 60 mm),

Gasterosteus aculeatus Ammonium

chloride 4 d S,M 7.5 23.3 78.7 142.8 Hazel et al. 1971

Threespine stickleback

(juvenile

- adult, 32

- 60 mm),

Gasterosteus aculeatus Ammonium

chloride 4 d S,M 7.5 23.3 115.4 209.5 Hazel et al. 1971

Threespine stickleback

(juvenile

- adult, 32

- 60 mm),

Gasterosteus aculeatus Ammonium

chloride 4 d S,M 7.5 15 259 470.0 281.5 281.5 Hazel et al. 1971

Mayfly,

Callibaetis skokianus Ammonium

chloride 4 d F,M 7.7 10.8 263.5 307.2 Arthur et al. 1987

Mayfly,

Callibaetis skokianus Ammonium

chloride 4 d F,M 7.9 13.3 211.7 432.7 364.6 West 1985; Arthur et al.

1987

Mayfly (middle to late instar),

Callibaetis sp. Ammonium

chloride 4 d F,M 7.81 11.9 107.8 166.7 166.7 246.5 Thurston et al. 1984b

## 89

Appendix A. Acute Toxicity of Ammonia to Aquatic Animals

Species Chemical Name Duration Methods a pH Temp.

( ̊C) Total

Ammonia

(mg

TA N/L) Total Ammonia b

(mg TA N/L)

a djusted to pH 7

(all organisms )

and 20 ° C

( invertebrates ) SMAV

(mg

TA N/L) GMAV

(mg

TA N/L) Reference

Dragonfly ( <233 d old),

Pachydiplax longipennis Ammonium

chloride 4 d F,M 8 12 76.92 170.1 Diamond et al. 1993

Dragonfly (<140 d old),

Pachydiplax longipennis Ammonium

chloride 4 d F,M 8 20 74.37 319.2 233.0 233.0 Diamond et al. 1993

Mottled sculpin (1.8 g, 5.4 cm),

Cottus bairdii Ammonium

chloride 4 d F,M 8.02 12.4 49.83 222.2 222.2 222.2 Thurston and Russo 1981

Western mosquitofish,

Gambusia affinis - 4 d S,U 7.75 19 129.6 352.9 Wallen et al. 1957

Western mosquitofish,

Gambusia affinis - 4 d S,U 8.2 19.5 34.54 217.7 Wallen et al. 1957

Western mosquitofish,

Gambusia affinis - 4 d S,U 8.5 23 14.64 165.0 Wallen et al. 1957

Western mosquitofish,

Gambusia affinis - 4 d S,U 8 24 42.53 182.6 219.3 219.3 Wallen et al. 1957

Oligochaete worm,

Lumbriculus variegatus Ammonium

chloride 4 d S,M 7.56 23 286 729.5 Besser et al. 1998

Oligochaete worm,

Lumbriculus variegatus Ammonium

chloride 4 d S,M 6.69 23 302 311.1 Besser et al. 1998

Oligochaete worm (10

- 25 mm),

Lumbriculus variegatus Ammonium

chloride 4 d R,M 8.2 15 13.66 56.88 Hickey and Vickers 1994

Oligochaete worm (adult),

Lumbriculus variegatus - 4 d F,M 6.5 25 100 111.9 Schubauer

- Berigan et al.

1995

Oligochaete worm (adult),

Lumbriculus variegatus - 4 d F,M 6.5 25 200 223.8 Schubauer

- Berigan et al.

1995

Oligochaete worm (adult),

Lumbriculus variegatus - 4 d F,M 8.1 25 34 267.3 Schubauer

- Berigan et al.

1995

Oligochaete worm (adult),

Lumbriculus variegatus - 4 d F,M 8.1 25 43.5 342.0 218.7 218.7 Schubauer

- Berigan et al.

1995

Tubificid worm,

Tubifex tubifex Ammonium

chloride 4 d S,U 8.2 12 66.67 216.5 216.5 216.5 Stammer 1953

## 90

Appendix A. Acute Toxicity of Ammonia to Aquatic Animals

Species Chemical Name Duration Methods a pH Temp.

( ̊C) Total

Ammonia

(mg

TA N/L) Total Ammonia b

(mg TA N/L)

a djusted to pH 7

(all organisms )

and 20 ° C

( invertebrates ) SMAV

(mg

TA N/L) GMAV

(mg

TA N/L) Reference

Marsh ramshorn snail,

Planorbella trivolvis

(previously Helisoma trivolvis ) Ammonium

chloride 4 d F,M 7.9 22 47.73 200.7 Arthur et al. 1987

Marsh ramshorn snail,

Planorbella trivolvis Ammonium

chloride 4 d F,M 8.2 12.9 63.73 223.0 211.6 211.6 Arthur et al. 1987

Scud (7

- 14 d old),

Hyalella azteca Ammonium

chloride 4 d R,M 8.3 25 39.8 461.2 Ankley et al. 1995

Scud (7

- 14 d old),

Hyalella azteca Ammonium

chloride 4 d R,M 7.31 25 64 135.1 Ankley et al. 1995

Scud (7

- 14 d old),

Hyalella azteca Ammonium

chloride 4 d R,M 6.43 25 105 114.6 192.6 192.6 Ankley et al. 1995

Stonefly, Little golden stonefly

(middle to late instar),

Skwala americana Ammonium

chloride 4 d F,M 7.81 13.1 109.3 186.7 Thurston et al. 1984b

Stonefly, Little golden stonefly

(middle to late instar),

Skwala americana Ammonium

chloride 4 d F,M 7.76 13.8 119.6 198.3 192.4 192.4 Thurston et al. 1984b

Mozambique tilapia (juvenile),

Oreochromis mossambicus Ammonium

chloride 4 d R,U 7.2 28 151.5 185.2 185.2 185.2 Rani et al. 1998

Amphipod (4

- 6 mm),

Crangonyx pseudogracilis Ammonium

chloride 4 d S,U 7.5 12 43.36 40.54 Prenter et al. 2004

Amphipod,

Crangonyx pseudogracilis Ammonium

chloride 4 d F,M 8 4 199.5 227.3 West 1985; Arthur et al.

1987

Amphipod,

Crangonyx pseudogracilis Ammonium

chloride 4 d F,M 8 12.1 216 481.7 West 1985; Arthur et al.

1987

Amphipod,

Crangonyx pseudogracilis Ammonium

chloride 4 d F,M 8 13.3 115.3 284.1 West 1985; Arthur et al.

1987

Amphipod,

Crangonyx pseudogracilis Ammonium

chloride 4 d F,M 8 24.9 25.1 161.7 West 1985; Arthur et al.

1987

Amphipod,

Crangonyx pseudogracilis Ammonium

chloride 4 d F,M 8.2 13 81.6 287.9 270.5 West 1985; Arthur et al.

1987

## 91

Appendix A. Acute Toxicity of Ammonia to Aquatic Animals

Species Chemical Name Duration Methods a pH Temp.

( ̊C) Total

Ammonia

(mg

TA N/L) Total Ammonia b

(mg TA N/L)

a djusted to pH 7

(all organisms )

and 20 ° C

( invertebrates ) SMAV

(mg

TA N/L) GMAV

(mg

TA N/L) Reference

Amphipod (13 d),

Crangonyx sp. Ammonium

chloride 4 d F,M 8 12 79.23 175.3 Diamond et al. 1993

Amphipod (8

- 42 d),

Crangonyx sp. Ammonium

chloride 4 d F,M 8 20 19.83 85.13 122.2 181.8 Diamond et al. 1993

Tubificid worm (30

- 40 mm),

Limnodrilus hoffmeisteri - 4 d F,M 7.9 11.5 96.62 170.2 170.2 170.2 Williams et al. 1986

Pouch snail,

Physa gyrina Ammonium

chloride 4 d F,M 8 4 114.9 131.0 West 1985; Arthur et al.

1987

Pouch snail,

Physa gyrina Ammonium

chloride 4 d F,M 8.2 5.5 85.13 161.3 West 1985; Arthur et al.

1987

Pouch snail,

Physa gyrina Ammonium

chloride 4 d F,M 8.1 12.1 76.29 205.9 West 1985; Arthur et al.

1987

Pouch snail,

Physa gyrina Ammonium

chloride 4 d F,M 8.2 12.8 50.25 174.4 West 1985; Arthur et al.

1987

Pouch snail,

Physa gyrina Ammonium

chloride 4 d F,M 8 13.3 62.39 153.7 West 1985; Arthur et al.

1987

Pouch snail,

Physa gyrina Ammonium

chloride 4 d F,M 8 24.9 26.33 169.7 164.5 164.5 West 1985; Arthur et al.

1987

Damselfly (8

- 10 mm),

Enallagma sp. - 4 d F,M 7.9 11.5 93.1 164.0 164.0 164.0 Williams et al. 1986

Water flea (<24 hr),

Chydorus sphaericus Ammonium

chloride 4 d S,M 8 20 37.88 162.6 162.6 162.6 Dekker et al. 2006

Fathead minnow (larva, 14 d),

Pimephales promelas - 4 d S,U 7.6 20 37.56 79.59 Markle et al. 2000

Fathead minnow,

Pimephales promelas Ammonium

chloride 4 d S,M 7.52 20.25 36.73 68.17 EA Engineering 1985

Fathead minnow,

Pimephales promelas Ammonium

chloride 4 d S,M 7.48 19.85 40.93 72.10 EA Engineering 1985

Fathead minnow,

Pimephales promelas Ammonium

chloride 4 d S,M 7.52 20.25 37.49 69.59 EA Engineering 1985

## 92

Appendix A. Acute Toxicity of Ammonia to Aquatic Animals

Species Chemical Name Duration Methods a pH Temp.

( ̊C) Total

Ammonia

(mg

TA N/L) Total Ammonia b

(mg TA N/L)

a djusted to pH 7

(all organisms )

and 20 ° C

( invertebrates ) SMAV

(mg

TA N/L) GMAV

(mg

TA N/L) Reference

Fathead minnow,

Pimephales promelas Ammonium

chloride 4 d S,M 7.48 19.85 41.79 73.61 EA Engineering 1985

Fathead minnow,

Pimephales promelas Ammonium

chloride 4 d S,M 7.48 19.85 43.49 76.61 EA Engineering 1985

Fathead minnow (4

- 6 d old),

Pimephales promelas Ammonium

chloride 4 d R,M 8.01 25 14.4 63.00 Buhl 2002

Fathead minnow,

Pimephales promelas Ammonium

chloride 4 d R,M 8 20 5.389 23.13 Diamond et al. 1993

Fathead minnow,

Pimephales promelas Ammonium

chloride 4 d R,M 8 20 6.1 26.19 Diamond et al. 1993

Fathead minnow (1.9 g),

Pimephales promelas Ammonium

chloride 4 d F,M 7.9 3.4 229.7 818.4 West 1985; Arthur et al.

1987

Fathead minnow (1.8 g),

Pimephales promelas Ammonium

chloride 4 d F,M 8.1 12.1 56.07 291.3 West 1985; Arthur et al.

1987

Fathead minnow (1.6 g),

Pimephales promelas Ammonium

chloride 4 d F,M 8 17.1 52.22 224.2 West 1985; Arthur et al.

1987

Fathead minnow (1.7 g),

Pimephales promelas Ammonium

chloride 4 d F,M 8.1 26.1 29.23 151.8 West 1985; Arthur et al.

1987

Fathead minnow,

Pimephales promelas Ammonium

chloride 4 d F,M 8.05 14 47.29 223.2 DeGraeve et al. 1980

Fathead minnow (4

- 5 mo),

Pimephales promelas Ammonium

chloride 4 d F,M 7.46 6 97.27 166.4 DeGraeve et al. 1987

Fathead minnow (4

- 5 mo),

Pimephales promelas Ammonium

chloride 4 d F,M 7.46 10 101.7 174.0 DeGraeve et al. 1987

Fathead minnow (4

- 5 mo),

Pimephales promelas Ammonium

chloride 4 d F,M 7.41 15 76.58 122.0 DeGraeve et al. 1987

Fathead minnow (4

- 5 mo),

Pimephales promelas Ammonium

chloride 4 d F,M 7.41 20 78.22 124.6 DeGraeve et al. 1987

Fathead minnow (4

- 5 mo),

Pimephales promelas Ammonium

chloride 4 d F,M 7.45 20 66.94 112.9 DeGraeve et al. 1987

Fathead minnow (4

- 5 mo),

Pimephales promelas Ammonium

chloride 4 d F,M 7.4 25 81.81 128.5 DeGraeve et al. 1987

Fathead minnow (4

- 5 mo),

Pimephales promelas Ammonium

chloride 4 d F,M 7.41 25 91.4 145.6 DeGraeve et al. 1987

Fathead minnow (4

- 5 mo),

Pimephales promelas Ammonium

chloride 4 d F,M 7.44 30 64.12 106.6 DeGraeve et al. 1987

## 93

Appendix A. Acute Toxicity of Ammonia to Aquatic Animals

Species Chemical Name Duration Methods a pH Temp.

( ̊C) Total

Ammonia

(mg

TA N/L) Total Ammonia b

(mg TA N/L)

a djusted to pH 7

(all organisms )

and 20 ° C

( invertebrates ) SMAV

(mg

TA N/L) GMAV

(mg

TA N/L) Reference

Fathead minnow

(0.28 g, 26.6 mm),

Pimephales promelas Ammonium

chloride 4 d F,M 8.14 22 25.16 141.2 Mayes et al. 1986

Fathead minnow

(10 mm length),

Pimephales promelas Ammonium

chloride 4 d F,M 7.9 20.6 28.9 103.0 Nimmo et al. 1989

Fathead minnow (10 mm),

Pimephales promelas Ammonium

chloride 4 d F,M 8.2 6.2 7.322 46.15 Nimmo et al. 1989

Fathead minnow (10 mm),

Pimephales promelas Ammonium

chloride 4 d F,M 7.8 20.1 18.73 55.68 Nimmo et al. 1989

Fathead minnow (10 mm),

Pimephales promelas Ammonium

chloride 4 d F,M 7.8 19.8 32.12 95.49 Nimmo et al. 1989

Fathead minnow (25 mm),

Pimephales promelas Ammonium

chloride 4 d F,M 8.1 19.6 24.89 129.3 Nimmo et al. 1989

Fathead minnow (25 mm),

Pimephales promelas Ammonium

chloride 4 d F,M 8.2 6.2 11.56 72.86 Nimmo et al. 1989

Fathead minnow (25 mm),

Pimephales promelas Ammonium

chloride 4 d F,M 8.1 5.8 19.94 103.6 Nimmo et al. 1989

Fathead minnow (25 mm),

Pimephales promelas Ammonium

chloride 4 d F,M 8.1 5.8 21.44 111.4 Nimmo et al. 1989

Fathead minnow (25 mm),

Pimephales promelas Ammonium

chloride 4 d F,M 7.7 20.1 32.25 80.61 Nimmo et al. 1989

Fathead minnow

(15 mm, 0.0301 g),

Pimephales promelas Ammonium

chloride 4 d F,M 8.46 4.1 18.54 193.5 Reinbold and Pescitelli

1982b

Fathead minnow

(16 mm, 0.0315 g),

Pimephales promelas Ammonium

chloride 4 d F,M 8.02 23.9 19.55 87.16 Reinbold and Pescitelli

1982b

Fathead minnow

(19 mm, 0.0629 g),

Pimephales promelas Ammonium

chloride 4 d F,M 8.26 4.6 30.57 216.5 Reinbold and Pescitelli

1982b

Fathead minnow

(21 mm, 0.0662 g),

Pimephales promelas Ammonium

chloride 4 d F,M 8.16 25.2 17.65 102.9 Reinbold and Pescitelli

1982b

## 94

Appendix A. Acute Toxicity of Ammonia to Aquatic Animals

Species Chemical Name Duration Methods a pH Temp.

( ̊C) Total

Ammonia

(mg

TA N/L) Total Ammonia b

(mg TA N/L)

a djusted to pH 7

(all organisms )

and 20 ° C

( invertebrates ) SMAV

(mg

TA N/L) GMAV

(mg

TA N/L) Reference

Fathead minnow

(5.2 cm, 1.1 g),

Pimephales promelas Ammonium

chloride 4 d F,M 7.7 21.65 63.02 157.5 Sparks 1975

Fathead minnow (0.2 g),

Pimephales promelas Ammonium

chloride 4 d F,M 7.78 25.9 40.85 117.3 Swigert and Spacie 1983

Fathead minnow (0.5 g),

Pimephales promelas Ammonium

chloride 4 d F,M 7.8 25.6 42.65 126.8 Swigert and Spacie 1983

Fathead minnow

(1.9 g, 5.2 cm),

Pimephales promelas Ammonium

chloride 4 d F,M 7.83 11.8 45.71 143.4 Thurston et al. 1981c

Fathead minnow

(1.9 g, 5.2 cm),

Pimephales promelas Ammonium

chloride 4 d F,M 7.82 12 62.72 193.3 Thurston et al. 1981c

Fathead minnow

(1.9 g, 5.2 cm),

Pimephales promelas Ammonium

chloride 4 d F,M 6.51 13 260 192.9 Thurston et al. 1981c

Fathead minnow

(1.9 g, 5.2 cm),

Pimephales promelas Ammonium

chloride 4 d F,M 9.03 13.2 5.94 169.6 Thurston et al. 1981c

Fathead minnow

(1.9 g, 5.2 cm),

Pimephales promelas Ammonium

chloride 4 d F,M 8.51 13.5 18.88 216.9 Thurston et al. 1981c

Fathead minnow

(1.9 g, 5.2 cm),

Pimephales promelas Ammonium

chloride 4 d F,M 7.01 13.8 145.9 147.2 Thurston et al. 1981c

Fathead minnow

(0.09 g, 2.0 cm),

Pimephales promelas Ammonium

chloride 4 d F,M 7.91 16.3 51.55 187.1 Thurston et al. 1983

Fathead minnow

(0.09 g, 2.1 cm),

Pimephales promelas Ammonium

chloride 4 d F,M 7.89 13.1 50.2 175.6 Thurston et al. 1983

Fathead minnow

(0.13 g, 2.3 cm),

Pimephales promelas Ammonium

chloride 4 d F,M 7.64 13.6 58.4 132.1 Thurston et al. 1983

## 95

Appendix A. Acute Toxicity of Ammonia to Aquatic Animals

Species Chemical Name Duration Methods a pH Temp.

( ̊C) Total

Ammonia

(mg

TA N/L) Total Ammonia b

(mg TA N/L)

a djusted to pH 7

(all organisms )

and 20 ° C

( invertebrates ) SMAV

(mg

TA N/L) GMAV

(mg

TA N/L) Reference

Fathead minnow

(0.19 g, 2.6 cm),

Pimephales promelas Ammonium

chloride 4 d F,M 7.68 13.5 64.7 156.3 Thurston et al. 1983

Fathead minnow

(0.22 g, 2.7 cm),

Pimephales promelas Ammonium

chloride 4 d F,M 8.03 22.1 47.6 216.3 Thurston et al. 1983

Fathead minnow

(0.22 g, 2.9 cm),

Pimephales promelas Ammonium

chloride 4 d F,M 8.06 22 42.6 205.0 Thurston et al. 1983

Fathead minnow

(0.26 g, 3.0 cm),

Pimephales promelas Ammonium

chloride 4 d F,M 7.67 13.9 58.8 139.7 Thurston et al. 1983

Fathead minnow

(0.31 g, 3.0 cm),

Pimephales promelas Ammonium

chloride 4 d F,M 8.05 13 74.65 352.4 Thurston et al. 1983

Fathead minnow

(0.31 g, 3.1 cm),

Pimephales promelas Ammonium

chloride 4 d F,M 8.05 13.6 66.48 313.8 Thurston et al. 1983

Fathead minnow

(0.35 g, 3.1 cm),

Pimephales promelas Ammonium

chloride 4 d F,M 7.94 19.1 42.3 162.3 Thurston et al. 1983

Fathead minnow

(0.42 g, 3.0 cm),

Pimephales promelas Ammonium

chloride 4 d F,M 7.76 19 50.28 139.3 Thurston et al. 1983

Fathead minnow

(0.42 g, 3.6 cm),

Pimephales promelas Ammonium

chloride 4 d F,M 7.66 13.4 58.2 136.0 Thurston et al. 1983

Fathead minnow

(0.47 g, 3.2 cm),

Pimephales promelas Ammonium

chloride 4 d F,M 7.87 15.8 58.91 198.7 Thurston et al. 1983

Fathead minnow

(0.47 g, 3.2 cm),

Pimephales promelas Ammonium

chloride 4 d F,M 7.83 22 50.6 158.7 Thurston et al. 1983

Fathead minnow

(0.5 g, 3.8 cm),

Pimephales promelas Ammonium

chloride 4 d F,M 7.91 18.9 49.3 178.9 Thurston et al. 1983

## 96

Appendix A. Acute Toxicity of Ammonia to Aquatic Animals

Species Chemical Name Duration Methods a pH Temp.

( ̊C) Total

Ammonia

(mg

TA N/L) Total Ammonia b

(mg TA N/L)

a djusted to pH 7

(all organisms )

and 20 ° C

( invertebrates ) SMAV

(mg

TA N/L) GMAV

(mg

TA N/L) Reference

Fathead minnow

(0.8 g, 4.2 cm),

Pimephales promelas Ammonium

chloride 4 d F,M 7.77 14.3 66.7 188.1 Thurston et al. 1983

Fathead minnow

(1.0 g, 4.6 cm),

Pimephales promelas Ammonium

chloride 4 d F,M 7.77 14.1 72.71 205.1 Thurston et al. 1983

Fathead minnow

(1.4 g, 4.9 cm),

Pimephales promelas Ammonium

chloride 4 d F,M 8.04 22.4 36.59 169.5 Thurston et al. 1983

Fathead minnow

(1.4 g, 5.0 cm),

Pimephales promelas Ammonium

chloride 4 d F,M 8.08 21.4 44.8 224.0 Thurston et al. 1983

Fathead minnow

(1.4 g, 5.0 cm),

Pimephales promelas Ammonium

chloride 4 d F,M 8.16 21.4 47.39 276.4 Thurston et al. 1983

Fathead minnow

(1.4 g, 5.1 cm),

Pimephales promelas Ammonium

chloride 4 d F,M 7.88 21.7 50.9 174.8 Thurston et al. 1983

Fathead minnow

(1.4 g, 5.4 cm),

Pimephales promelas Ammonium

chloride 4 d F,M 7.68 12.9 91.8 221.8 Thurston et al. 1983

Fathead minnow

(1.4 g, 5.5 cm),

Pimephales promelas Ammonium

chloride 4 d F,M 7.63 13.2 89.85 199.9 Thurston et al. 1983

Fathead minnow

(1.5 g, 5.6 cm),

Pimephales promelas Ammonium

chloride 4 d F,M 7.76 12.9 107.5 298.0 Thurston et al. 1983

Fathead minnow

(1.7 g, 5.2 cm),

Pimephales promelas Ammonium

chloride 4 d F,M 7.84 21.7 55.43 177.0 Thurston et al. 1983

Fathead minnow

(2.1 g, 6.1 cm),

Pimephales promelas Ammonium

chloride 4 d F,M 7.76 13.1 66.73 184.9 Thurston et al. 1983

Fathead minnow

(2.2 g, 6.2 cm),

Pimephales promelas Ammonium

chloride 4 d F,M 7.74 12.8 52.2 139.7 Thurston et al. 1983

## 97

Appendix A. Acute Toxicity of Ammonia to Aquatic Animals

Species Chemical Name Duration Methods a pH Temp.

( ̊C) Total

Ammonia

(mg

TA N/L) Total Ammonia b

(mg TA N/L)

a djusted to pH 7

(all organisms )

and 20 ° C

( invertebrates ) SMAV

(mg

TA N/L) GMAV

(mg

TA N/L) Reference

Fathead minnow

(2.3 g, 6.3 cm),

Pimephales promelas Ammonium

chloride 4 d F,M 7.91 15.9 47.43 172.1 159.2 159.2 Thurston et al. 1983

Brook trout (3.12 g, 7.2 cm),

Salvelinus fontinalis Ammonium

chloride 4 d F,U 7.86 13.6 45.21 149.7 Thurston and Meyn 1984

Brook trout (3.40 g, 7.4 cm),

Salvelinus fontinalis Ammonium

chloride 4 d F,U 7.83 13.8 52.03 163.2 156.3 Thurston and Meyn 1984

Lake trout, siscowet (0.9 g),

Salvelinus namaycush Ammonium

chloride 4 d S,M 7.45 8.5 90.43 152.5 Soderberg and Meade 1992

Lake trout, siscowet (0.9 g),

Salvelinus namaycush Ammonium

chloride 4 d S,M 7.45 8.5 110.2 185.9 Soderberg and Meade 1992

Lake trout, siscowet (8 g),

Salvelinus namaycush Ammonium

chloride 4 d S,M 7.45 8.5 96.25 162.3 Soderberg and Meade 1992

Lake trout, siscowet (8 g),

Salvelinus namaycush Ammonium

chloride 4 d S,M 7.45 8.5 83.11 140.1 159.3 157.8 Soderberg and Meade 1992

Shortnose sturgeon (fingerling),

Acipenser brevirostrum Ammonium

chloride 4 d S,M 7.05 18 149.8 156.7 156.7 156.7 Fontenot et al. 1998

White sucker (5.6 g),

Catostomus commersonii Ammonium

chloride 4 d F,M 7.8 3.6 89.57 266.3 West 1985; Arthur et al.

1987

White sucker (5.2 g),

Catostomus commersonii Ammonium

chloride 4 d F,M 8.1 11.3 60.86 316.1 West 1985; Arthur et al.

1987

White sucker (12.6 g),

Catostomus commersonii Ammonium

chloride 4 d F,M 8.2 12.6 40.85 257.4 West 1985; Arthur et al.

1987

White sucker (9.6 g),

Catostomus commersonii Ammonium

chloride 4 d F,M 8.2 15.3 43.01 271.0 West 1985; Arthur et al.

1987

White sucker (110 mm),

Catostomus commersonii Ammonium

chloride 4 d F,M 7.8 20.2 31.21 92.80 Nimmo et al. 1989

White sucker (110 mm),

Catostomus commersonii Ammonium

chloride 4 d F,M 7.8 20.2 18.93 56.28 Nimmo et al. 1989

White sucker (92 mm, 6.3 g),

Catostomus commersonii Ammonium

chloride 4 d F,M 8.16 15 30.28 176.6 Reinbold and Pescitelli

1982c

## 98

Appendix A. Acute Toxicity of Ammonia to Aquatic Animals

Species Chemical Name Duration Methods a pH Temp.

( ̊C) Total

Ammonia

(mg

TA N/L) Total Ammonia b

(mg TA N/L)

a djusted to pH 7

(all organisms )

and 20 ° C

( invertebrates ) SMAV

(mg

TA N/L) GMAV

(mg

TA N/L) Reference

White sucker (92 mm, 6.3 g),

Catostomus commersonii Ammonium

chloride 4 d F,M 8.14 15.4 29.65 166.3 Reinbold and Pescitelli

1982c

White sucker (11.4 g),

Catostomus commersonii Ammonium

chloride 4 d F,M 7.8 22.5 22.3 66.32 157.5 Swigert and Spacie 1983

Mountain sucker

(63.3 g, 18.2 cm),

Catostomus platyrhynchus Ammonium

chloride 4 d F,U 7.67 12 66.91 159.0 Thurston and Meyn 1984

Mountain sucker

(45.3 g, 16.2 cm),

Catostomus platyrhynchus Ammonium

chloride 4 d F,U 7.69 13.2 47.59 117.0 Thurston and Meyn 1984

Mountain sucker

(47.8 g, 15.9 cm),

Catostomus platyrhynchus Ammonium

chloride 4 d F,U 7.73 11.7 51.62 135.8 136.2 146.5 Thurston and Meyn 1984

Water flea,

Ceriodaphnia acanthin a Ammonium

chloride 2 d F,M 7.06 24 104.8 154.3 154.3 Mount 1982

Water flea (<24 hr),

Ceriodaphnia dubia Ammonium

chloride 2 d S,M 8.02 24.8 21.26 141.1 Andersen and Buckley

1998

Water flea (<24 hr),

Ceriodaphnia dubia Ammonium

chloride 2 d S,M 7.5 25 47.05 129.2 Bailey et al. 2001

Water flea (<24 hr),

Ceriodaphnia dubia Ammonium

chloride 2 d S,M 7.5 25 56.84 156.1 Bailey et al. 2001

Water flea (<24 hr),

Ceriodaphnia dubia Ammonium

chloride 2 d S,M 8.16 22 24.77 170.5 Black 2001

Water flea (<24 hr),

Ceriodaphnia dubia Ammonium

chloride 2 d S,M 8.4 23 28.06 334.5 Black 2001

Water flea (<24 hr),

Ceriodaphnia dubia Ammonium

chloride 2 d S,M 8.4 23 32.63 389.0 Black 2001

Water flea (<24 hr),

Ceriodaphnia dubia Ammonium

chloride 2 d S,M 8 25 14.52 94.35 Scheller 1997

Water flea,

Ceriodaphnia dubia Ammonium

chloride 2 d S,M 8.08 24.75 15.6 114.5 Andersen and Buckley

1998

Water flea,

Ceriodaphnia dubia Ammonium

hydroxide 2 d R,M 8.4 26.4 7.412 117.1 Cowgill and Milazzo 1991

## 99

Appendix A. Acute Toxicity of Ammonia to Aquatic Animals

Species Chemical Name Duration Methods a pH Temp.

( ̊C) Total

Ammonia

(mg

TA N/L) Total Ammonia b

(mg TA N/L)

a djusted to pH 7

(all organisms )

and 20 ° C

( invertebrates ) SMAV

(mg

TA N/L) GMAV

(mg

TA N/L) Reference

Water flea,

Ceriodaphnia dubia Ammonium

sulfate 2 d R,NR 7.4 23 48.59 97.89 Manning et al. 1996

Water flea,

Ceriodaphnia dubia Ammonium

chloride 2 d R,M 7.8 25 33.98 152.9 Nimmo et al. 1989

Water flea,

Ceriodaphnia dubia Ammonium

chloride 2 d R,M 8.2 7 16.65 35.72 Nimmo et al. 1989

Water flea,

Ceriodaphnia dubia Ammonium

chloride 2 d S,M 7.85 23 28.65 119.5 Sarda 1994

Water flea,

Ceriodaphnia dubia Ammonium

chloride 2 d S,M 7.85 23 28.77 120.0 134.2 143.9 Sarda 1994

Water flea (adult),

Simocephalus vetulus Ammonium

chloride 2 d F,M 8.3 17 31.58 188.5 West 1985; Arthur et al.

1987

Water flea (adult),

Simocephalus vetulus Ammonium

chloride 2 d F,M 8.1 20.4 21.36 114.7 Arthur et al. 1987

Water flea,

Simocephalus vetulus Ammonium

chloride 2 d F,M 7.25 24.5 83.51 157.0 Mount 1982

Water flea,

Simocephalus vetulus Ammonium

chloride 2 d F,M 7.06 24 83.51 122.9 142.9 142.9 Mount 1982

Channel catfish,

Ictalurus punctatus Ammonium

chloride 4 d S,U 8.7 22 10.56 172.9 Colt and Tchobanoglous

1976

Channel catfish,

Ictalurus punctatus Ammonium

chloride 4 d S,U 8.7 26 10.19 166.9 Colt and Tchobanoglous

1976

Channel catfish,

Ictalurus punctatus Ammonium

chloride 4 d S,U 8.7 30 10.88 178.1 Colt and Tchobanoglous

1976

Channel catfish,

Ictalurus punctatus Ammonium

chloride 4 d S,M 7.49 19.7 131.5 235.0 EA Engineering 1985

Channel catfish,

Ictalurus punctatus Ammonium

chloride 4 d S,M 7.53 19.75 99.67 189.3 EA Engineering 1985

Channel catfish (larvae, 1 d),

Ictalurus punctatus Ammonium

chloride 4 d R,M 8.2 23.8 13.03 82.10 Bader and Grizzle 1992

Channel catfish (juvenile, 7 d),

Ictalurus punctatus Ammonium

chloride 4 d R,M 8.2 23.9 17.22 108.5 Bader and Grizzle 1992

Channel catfish (3.5 g),

Ictalurus punctatus Ammonium

chloride 4 d F,M 7.8 19.6 44.71 132.9 West 1985; Arthur et al.

1987

## 100

Appendix A. Acute Toxicity of Ammonia to Aquatic Animals

Species Chemical Name Duration Methods a pH Temp.

( ̊C) Total

Ammonia

(mg

TA N/L) Total Ammonia b

(mg TA N/L)

a djusted to pH 7

(all organisms )

and 20 ° C

( invertebrates ) SMAV

(mg

TA N/L) GMAV

(mg

TA N/L) Reference

Channel catfish (5.8 g),

Ictalurus punctatus Ammonium

chloride 4 d F,M 8 3.5 37.64 161.6 West 1985; Arthur et al.

1987

Channel catfish (6.4 g),

Ictalurus punctatus Ammonium

chloride 4 d F,M 8.1 14.6 24.94 129.5 West 1985; Arthur et al.

1987

Channel catfish,

Ictalurus punctatus Ammonium

chloride 4 d F,M 8.4 28 10.71 99.59 Colt and Tchobanoglous

1978

Channel catfish (3

- 11 mo),

Ictalurus punctatus Ammonium

chloride 4 d F,M 7.46 10 124.8 213.5 DeGraeve et al. 1987

Channel catfish (3

- 11 mo),

Ictalurus punctatus Ammonium

chloride 4 d F,M 7.41 15 113.1 180.2 DeGraeve et al. 1987

Channel catfish (3

- 11 mo),

Ictalurus punctatus Ammonium

chloride 4 d F,M 7.41 20 89.63 142.8 DeGraeve et al. 1987

Channel catfish (3

- 11 mo),

Ictalurus punctatus Ammonium

chloride 4 d F,M 7.45 20 72.15 121.7 DeGraeve et al. 1987

Channel catfish (3

- 11 mo),

Ictalurus punctatus Ammonium

chloride 4 d F,M 7.4 25 89.41 140.5 DeGraeve et al. 1987

Channel catfish (3

- 11 mo),

Ictalurus punctatus Ammonium

chloride 4 d F,M 7.41 25 85.69 136.5 DeGraeve et al. 1987

Channel catfish (3

- 11 mo),

Ictalurus punctatus Ammonium

chloride 4 d F,M 7.44 30 65.25 108.5 DeGraeve et al. 1987

Channel catfish (<110 d),

Ictalurus punctatus Ammonium

chloride 4 d F,M 8 20 15.09 64.77 Diamond et al. 1993

Channel catfish,

Ictalurus punctatus Ammonium

chloride 4 d F,M 7.94 23.8 33.1 127.0 Reinbold and Pescitelli

1982d

Channel catfish,

Ictalurus punctatus Ammonium

chloride 4 d F,M 7.98 23.8 30.49 126.1 Reinbold and Pescitelli

1982d

Channel catfish (4.5

- 10.8 g),

Ictalurus punctatus Ammonium

chloride 4 d F,M 8.08 28 44.44 222.2 Roseboom and Richey 1977

Channel catfish (7.1

- 12.7 g),

Ictalurus punctatus Ammonium

chloride 4 d F,M 8.09 22 32.33 164.8 Roseboom and Richey 1977

Channel catfish

(14.3 mm, 19.0 g),

Ictalurus punctatus Ammonium

chloride 4 d F,M 7.93 20 74.35 277.4 Sparks 1975

Channel catfish (0.5 g),

Ictalurus punctatus Ammonium

chloride 4 d F,M 7.8 25.7 32.85 97.67 Swigert and Spacie 1983

## 101

Appendix A. Acute Toxicity of Ammonia to Aquatic Animals

Species Chemical Name Duration Methods a pH Temp.

( ̊C) Total

Ammonia

(mg

TA N/L) Total Ammonia b

(mg TA N/L)

a djusted to pH 7

(all organisms )

and 20 ° C

( invertebrates ) SMAV

(mg

TA N/L) GMAV

(mg

TA N/L) Reference

Channel catfish,

Ictalurus punctatus Ammonium

chloride 4 d F,M 8 26 32.34 138.8 West 1985

Channel catfish,

Ictalurus punctatus Ammonium

chloride 4 d F,M 8.1 17 40.83 212.1 142.4 142.4 West 1985

Red swamp crayfish (2.1 cm),

Procambarus clarkii Ammonium

chloride 4 d F,M 8 20 26.08 112.0 Diamond et al. 1993

Red swamp crayfish (<2.5 cm),

Procambarus clarkii Ammonium

chloride 4 d F,M 8 12 76.92 170.1 138.0 138.0 Diamond et al. 1993

Atlantic salmon (4.8

- 9.2 cm),

Salmo salar Ammonium

sulfate 4 d S,M 6.4 1.8 123 87.86 Knoph 1992

Atlantic salmon (4.8

- 9.2 cm),

Salmo salar Ammonium

sulfate 4 d S,M 6.4 1.8 133.9 95.64 Knoph 1992

Atlantic salmon (4.8

- 9.2 cm),

Salmo salar Ammonium

sulfate 4 d S,M 6 2.1 297.2 195.1 Knoph 1992

Atlantic salmon (4.8

- 9.2 cm),

Salmo salar Ammonium

sulfate 4 d S,M 6 2.1 341.1 223.9 Knoph 1992

Atlantic salmon (4.8

- 9.2 cm),

Salmo salar Ammonium

sulfate 4 d S,M 6.05 2.5 400 264.4 Knoph 1992

Atlantic salmon (4.8

- 9.2 cm),

Salmo salar Ammonium

sulfate 4 d S,M 6.05 2.5 491.7 325.0 Knoph 1992

Atlantic salmon (4.8

- 9.2 cm),

Salmo salar Ammonium

sulfate 4 d S,M 6 7.3 581.5 381.7 Knoph 1992

Atlantic salmon (4.8

- 9.2 cm),

Salmo salar Ammonium

sulfate 4 d S,M 6 7.3 587.6 385.7 Knoph 1992

Atlantic salmon (4.8

- 9.2 cm),

Salmo salar Ammonium

sulfate 4 d S,M 6.45 7.4 171.3 124.4 Knoph 1992

Atlantic salmon (4.8

- 9.2 cm),

Salmo salar Ammonium

sulfate 4 d S,M 6.45 7.4 214.4 155.7 Knoph 1992

Atlantic salmon (4.8

- 9.2 cm),

Salmo salar Ammonium

sulfate 4 d S,M 6.45 12.5 230.6 167.4 Knoph 1992

Atlantic salmon (4.8

- 9.2 cm),

Salmo salar Ammonium

sulfate 4 d S,M 6.45 12.5 248.3 180.3 Knoph 1992

Atlantic salmon (4.8

- 9.2 cm),

Salmo salar Ammonium

sulfate 4 d S,M 6.05 12.5 403.5 266.7 Knoph 1992

## 102

Appendix A. Acute Toxicity of Ammonia to Aquatic Animals

Species Chemical Name Duration Methods a pH Temp.

( ̊C) Total

Ammonia

(mg

TA N/L) Total Ammonia b

(mg TA N/L)

a djusted to pH 7

(all organisms )

and 20 ° C

( invertebrates ) SMAV

(mg

TA N/L) GMAV

(mg

TA N/L) Reference

Atlantic salmon (4.8

- 9.2 cm),

Salmo salar Ammonium

sulfate 4 d S,M 6.05 12.5 451.5 298.5 Knoph 1992

Atlantic salmon (4.8

- 9.2 cm),

Salmo salar Ammonium

sulfate 4 d S,M 6.05 17.1 356.1 235.4 Knoph 1992

Atlantic salmon (4.8

- 9.2 cm),

Salmo salar Ammonium

sulfate 4 d S,M 6.05 17.1 373 246.6 Knoph 1992

Atlantic salmon (1.5 g),

Salmo salar Ammonium

chloride 4 d S,M 7.45 8.5 60.29 101.7 Soderberg and Meade 1992

Atlantic salmon (1.5 g),

Salmo salar Ammonium

chloride 4 d S,M 7.45 8.5 35.74 60.26 Soderberg and Meade 1992

Atlantic salmon (36 g),

Salmo salar Ammonium

chloride 4 d S,M 7.45 8.5 118.2 199.3 Soderberg and Meade 1992

Atlantic salmon (36 g),

Salmo salar Ammonium

chloride 4 d S,M 7.45 8.5 70.62 119.1 183.3 Soderberg and Meade 1992

Brown trout (1.20 g, 5.4 cm),

Salmo trutta Ammonium

chloride 4 d F,U 7.85 13.2 29.58 96.20 Thurston and Meyn 1984

Brown trout (1.17 g, 5.3 cm),

Salmo trutta Ammonium

chloride 4 d. F,U 7.86 13.8 32.46 107.5 Thurston and Meyn 1984

Brown trout (0.91 g, 4.9 cm),

Salmo trutta Ammonium

chloride 4 d F,U 7.82 14.2 33.3 102.6 102.0 136.7 Thurston and Meyn 1984

White perch (76 mm),

Morone americana Ammonium

chloride 4 d S,M 8 16 14.93 64.09 Stevenson 1977

White perch (76 mm),

Morone americana Ammonium

chloride 4 d S,M 6 16 418.4 274.7 132.7 Stevenson 1977

White bass (4.4 g),

Morone chrysops Ammonium

chloride 4 d S,M 7.09 19.7 132.4 144.0 144.0 Ashe et al. 1996

Striped bass (20

- 93 mm),

Morone saxatilis Ammonium

chloride 4 d S,M 7.4 23.3 92.17 144.8 Hazel et al. 1971

Striped bass (20

- 93 mm),

Morone saxatilis Ammonium

chloride 4 d S,M 7.5 23.3 73.45 133.3 Hazel et al. 1971

Striped bass (20

- 93 mm),

Morone saxatilis Ammonium

chloride 4 d S,M 7.35 15 259.7 378.9 Hazel et al. 1971

## 103

Appendix A. Acute Toxicity of Ammonia to Aquatic Animals

Species Chemical Name Duration Methods a pH Temp.

( ̊C) Total

Ammonia

(mg

TA N/L) Total Ammonia b

(mg TA N/L)

a djusted to pH 7

(all organisms )

and 20 ° C

( invertebrates ) SMAV

(mg

TA N/L) GMAV

(mg

TA N/L) Reference

Striped bass (20

- 93 mm),

Morone saxatilis Ammonium

chloride 4 d S,M 7.5 15 182.3 330.7 Hazel et al. 1971

Striped bass (20

- 93 mm),

Morone saxatilis Ammonium

chloride 4 d S,M 7.93 23.3 48.03 180.8 Hazel et al. 1971

Striped bass (20

- 93 mm),

Morone saxatilis Ammonium

chloride 4 d S,M 7.5 23.3 125.9 228.5 Hazel et al. 1971

Striped bass (20

- 93 mm),

Morone saxatilis Ammonium

chloride 4 d S,M 7.84 15 165.7 524.6 Hazel et al. 1971

Striped bass (20

- 93 mm),

Morone saxatilis Ammonium

chloride 4 d S,M 7.5 15 354.9 644.0 Hazel et al. 1971

Striped bass (126.6 g),

Morone saxatilis Ammonium

chloride 4 d S,M 8.3 21 12.86 98.43 246.2 Oppenborn and Goudie

1993

Sunshine bass (larvae, 12 h),

Morone saxatilis x chrysops Ammonium

chloride 4 d S,M 8.5 18.7 3.903 43.99 Harcke and Daniels 1999

Sunshine bass (367.2 g),

Morone saxatilis x chrysops Ammonium

chloride 4 d S,M 8.3 21 8.147 62.37 Oppenborn and Goudie

1993

Sunshine bass (42.7 g),

Morone saxatilis x chrysops Ammonium

chloride 4 d S,M 7 25 63.62 63.62 Weirich et al. 1993

Sunshine bass (42.7 g),

Morone saxatilis x chrysops Ammonium

chloride 4 d S,M 7 25 83.06 83.06 Weirich et al. 1993

Sunshine bass (42.7 g),

Morone saxatilis x chrysops Ammonium

chloride 4 d S,M 7 25 56.55 56.55 Weirich et al. 1993

Sunshine bass (42.7 g),

Morone saxatilis x chrysops Ammonium

chloride 4 d S,M 7 25 65.39 65.39 Weirich et al. 1993

Sunshine bass (42.7 g),

Morone saxatilis x chrysops Ammonium

chloride 4 d S,M 7 25 60.09 60.09 Weirich et al. 1993

Sunshine bass (42.7 g),

Morone saxatilis x chrysops Ammonium

chloride 4 d S,M 7 25 64.51 64.51 Weirich et al. 1993

Sunshine bass (42.7 g),

Morone saxatilis x chrysops Ammonium

chloride 4 d S,M 7 25 79.53 79.53 Weirich et al. 1993

Sunshine bass (42.7 g),

Morone saxatilis x chrysops Ammonium

chloride 4 d S,M 7 25 86.6 86.60 Weirich et al. 1993

Sunshine bass (42.7 g),

Morone saxatilis x chrysops Ammonium

chloride 4 d S,M 7 25 95.43 95.43 Weirich et al. 1993

## 104

Appendix A. Acute Toxicity of Ammonia to Aquatic Animals

Species Chemical Name Duration Methods a pH Temp.

( ̊C) Total

Ammonia

(mg

TA N/L) Total Ammonia b

(mg TA N/L)

a djusted to pH 7

(all organisms )

and 20 ° C

( invertebrates ) SMAV

(mg

TA N/L) GMAV

(mg

TA N/L) Reference

Sunshine bass (42.7 g),

Morone saxatilis x chrysops Ammonium

chloride 4 d S,M 7 25 105.2 105.2 70.22 134.8 Weirich et al. 1993

Water flea,

Daphnia magna Ammonium

chloride 2 d S,M 8.5 20 26.34 296.9 Gersich and Hopkins 1986

Water flea,

Daphnia magna Ammonium

chloride 2 d S,M 7.92 21 9.463 37.66 Gulyas and Fleit 1990

Water flea,

Daphnia magna Ammonium

chloride 2 d S,M 8.2 25 20.71 197.5 Parkhurst et al. 1979, 1981

Water flea,

Daphnia magna Ammonium

chloride 2 d R,U 8.34 19.7 51.92 419.1 Reinbold and Pescitelli

1982a

Water flea,

Daphnia magna Ammonium

chloride 2 d S,M 8.07 19.6 51.09 242.4 Russo et al. 1985

Water flea,

Daphnia magna Ammonium

chloride 2 d S,M 7.51 20.1 48.32 89.74 Russo et al. 1985

Water flea,

Daphnia magna Ammonium

chloride 2 d S,M 7.53 20.1 55.41 106.1 Russo et al. 1985

Water flea,

Daphnia magna Ammonium

chloride 2 d S,M 7.5 20.3 43.52 80.98 Russo et al. 1985

Water flea,

Daphnia magna Ammonium

chloride 2 d S,M 7.4 20.6 42.31 69.88 Russo et al. 1985

Water flea,

Daphnia magna Ammonium

chloride 2 d S,M 8.09 20.9 41.51 227.9 Russo et al. 1985

Water flea,

Daphnia magna Ammonium

chloride 2 d S,M 7.95 22 51.3 236.7 Russo et al. 1985

Water flea,

Daphnia magna Ammonium

chloride 2 d S,M 8.15 22 37.44 252.8 Russo et al. 1985

Water flea,

Daphnia magna Ammonium

chloride 2 d S,M 8.04 22.8 38.7 226.1 157.7 Russo et al. 1985

Water flea,

Daphnia pulicaria Ammonium

chloride 2 d F,M 8.05 14 34.5 99.03 99.03 125.0 DeGraeve et al. 1980

Clawed toad (embryo),

Xenopus laevis Ammonium

sulfate 4 d R,M 7.2 22 38.59 40.91 Schuytema and Nebeker

1999a

## 105

Appendix A. Acute Toxicity of Ammonia to Aquatic Animals

Species Chemical Name Duration Methods a pH Temp.

( ̊C) Total

Ammonia

(mg

TA N/L) Total Ammonia b

(mg TA N/L)

a djusted to pH 7

(all organisms )

and 20 ° C

( invertebrates ) SMAV

(mg

TA N/L) GMAV

(mg

TA N/L) Reference

Clawed toad (embryo),

Xenopus laevis Ammonium

sulfate 4 d R,M 7.2 22 119.6 126.8 Schuytema and Nebeker

1999a

Clawed toad (embryo),

Xenopus laevis Ammonium

nitrate 4 d R,M 7.2 24 32.37 39.55 Schuytema and Nebeker

1999a

Clawed toad (embryo),

Xenopus laevis Ammonium

sulfate 4 d R,M 7.2 24 60.71 74.17 Schuytema and Nebeker

1999a

Clawed toad

(17 mg, Gosner Stage 26

- 27),

Xenopus laevis Nitric acid

ammonium salt 4 d R,M 7.15 22 101.4 117.2 Schuytema and Nebeker

1999b

Clawed toad

(17 mg, Gosner Stage 26

- 27),

Xenopus laevis Ammonium

sulfate 4 d R,M 7.15 22 135.9 157.2 Schuytema and Nebeker

1999b

Clawed toad

(21 mg, Gosner Stage 26

- 27),

Xenopus laevis Ammonium

chloride 4 d R,M 7.15 22 128.3 148.4 Schuytema and Nebeker

1999b

Clawed toad (embryo),

Xenopus laevis Ammonium

phosphate 4 d R,M 8.43 25 37.3 367.4 Tietge et al. 2000

Clawed toad (embryo),

Xenopus laevis Ammonium

phosphate 4 d R,M 8.62 25 28.7 405.6 122.5 122.5 Tietge et al. 2000

Flatworm,

Dendrocoelum lacteum Ammonium

chloride 4 d S,U 8.2 18 22.37 119.5 119.5 119.5 Stammer 1953

Walleye,

Sander vitreus Ammonium

chloride 4 d F,U 8.08 18.2 17.43 87.13 Reinbold and Pescitelli

1982a

Walleye (22.6 g),

Sander vitreus Ammonium

chloride 4 d F,M 7.9 3.7 48.37 172.3 West 1985; Arthur et al.

1987

Walleye (19.4 g),

Sander vitreus Ammonium

chloride 4 d F,M 7.7 11.1 89.93 224.8 West 1985; Arthur et al.

1987

Walleye (13.4 g),

Sander vitreus Ammonium

chloride 4 d F,M 8.3 19 6.123 46.87 West 1985; Arthur et al.

1987

Walleye (3.0 g, 65.6 mm),

Sander vitreus Ammonium

chloride 4 d F,M 8.06 21.5 21.49 103.4 117.1 117.1 Mayes et al. 1986

Central stoneroller (2.1 g),

Campostoma anomalum Ammonium

chloride 4 d F,M 7.8 25.7 38.97 115.9 115.9 115.9 Swigert and Spacie 1983

## 106

Appendix A. Acute Toxicity of Ammonia to Aquatic Animals

Species Chemical Name Duration Methods a pH Temp.

( ̊C) Total

Ammonia

(mg

TA N/L) Total Ammonia b

(mg TA N/L)

a djusted to pH 7

(all organisms )

and 20 ° C

( invertebrates ) SMAV

(mg

TA N/L) GMAV

(mg

TA N/L) Reference

Rainbow dace,

Cyprinella lutrensis Ammonium

chloride 4 d F,M 8.3 24 24.37 186.5 Hazel et al. 1979

Rainbow dace,

Cyprinella lutrensis Ammonium

chloride 4 d F,M 9.1 24 6.502 206.1 196.1 Hazel et al. 1979

Spotfin shiner (31

- 85 mm),

Cyprinella spiloptera Ammonium

chloride 4 d F,M 7.95 26.5 18.52 72.39 Rosage et al. 1979

Spotfin shiner (41

- 78 mm),

Cyprinella spiloptera Ammonium

chloride 4 d F,M 8.15 26.5 16.27 93.07 Rosage et al. 1979

Spotfin shiner (0.5 g),

Cyprinella spiloptera Ammonium

chloride 4 d F,M 7.9 25.7 24.52 87.36 83.8 0 Swigert and Spacie 1983

Steelcolor shiner (0.5 g),

Cyprinella whipplei Ammonium

chloride 4 d F,M 7.9 25.7 22.72 80.94 80.94 110.0 Swigert and Spacie 1983

Dwarf wedgemussel

(glochidia),

Alasmidonta heterodon Ammonium

chloride 1 d S,M 8.3 20 >14.24 c >109.0 >109.0 >109.0 Wang et al. 2007b

Pink papershell (glochidia),

Potamilus ohiensis Ammonium

chloride 1 d S,M 8.3 20 >14.24 c >109.0 >109.0 >109.0 Wang et al. 2007b

Green sunfish

(larvae, 9 d swim up fry),

Lepomis cyanellus Ammonium

chloride 4 d F,U 8.28 26.2 8.43 62.07 Reinbold and Pescitelli

1982a

Green sunfish,

Lepomis cyanellus Ammonium

chloride 4 d F,M 7.84 12.3 33.09 105.7 Jude 1973

Green sunfish (62.5 mg),

Lepomis cyanellus Ammonium

chloride 4 d F,M 7.2 22.4 142.9 174.5 McCormick et al. 1984

Green sunfish (62.5 mg),

Lepomis cyanellus Ammonium

chloride 4 d F,M 6.61 22.4 254.5 197.0 McCormick et al. 1984

Green sunfish (62.5 mg),

Lepomis cyanellus Ammonium

chloride 4 d F,M 7.72 22.4 55.79 144.3 McCormick et al. 1984

Green sunfish (62.5 mg),

Lepomis cyanellus Ammonium

chloride 4 d F,M 8.69 22.4 9.24 148.6 150.8 McCormick et al. 1984

## 107

Appendix A. Acute Toxicity of Ammonia to Aquatic Animals

Species Chemical Name Duration Methods a pH Temp.

( ̊C) Total

Ammonia

(mg

TA N/L) Total Ammonia b

(mg TA N/L)

a djusted to pH 7

(all organisms )

and 20 ° C

( invertebrates ) SMAV

(mg

TA N/L) GMAV

(mg

TA N/L) Reference

Pumpkinseed (4.13

- 9.22 g),

Lepomis gibbosus Ammonium

chloride 4 d F,M 7.77 12 9.11 25.69 Jude 1973

Pumpkinseed,

Lepomis gibbosus Ammonium

chloride 4 d F,M 7.77 14 48.09 135.6 Thurston 1981

Pumpkinseed,

Lepomis gibbosus Ammonium

chloride 4 d F,M 7.77 14.5 42.02 118.5 Thurston 1981

Pumpkinseed,

Lepomis gibbosus Ammonium

chloride 4 d F,M 7.71 15.7 48.54 87.54 77.53 Thurston 1981

Bluegill,

Lepomis macrochirus Ammonium

chloride 4 d S,M 7.51 20.35 40.41 73.88 EA Engineering 1985

Bluegill,

Lepomis macrochirus Ammonium

chloride 4 d S,M 7.51 20.35 41.96 76.72 EA Engineering 1985

Bluegill,

Lepomis macrochirus Ammonium

chloride 4 d S,M 7.52 20.65 41.9 78.36 EA Engineering 1985

Bluegill,

Lepomis macrochirus Ammonium

chloride 4 d S,M 7.51 20.35 44.3 80.98 EA Engineering 1985

Bluegill,

Lepomis macrochirus Ammonium

chloride 4 d S,M 7.52 20.65 42.63 79.73 EA Engineering 1985

Bluegill,

Lepomis macrochirus Ammonium

chloride 4 d S,M 7.52 20.65 44.1 82.48 EA Engineering 1985

Bluegill (1.7 cm),

Lepomis macrochirus Ammonium

chloride 4 d F,M 8 20 21.56 92.54 Diamond et al. 1993

Bluegill,

Lepomis macrochirus Ammonium

chloride 4 d F,M 8 12 25.12 107.9 Diamond et al. 1993

Bluegill

(20.0

- 70.0 mm, young of year),

Lepomis macrochirus Ammonium

chloride 4 d F,M 8.11 18.5 16.73 88.57 Emery and Welch 1969

Bluegill

(20.0

- 70.0 mm, young of year),

Lepomis macrochirus Ammonium

chloride 4 d F,M 8.24 18.5 42.01 286.1 Emery and Welch 1969

Bluegill

(20.0

- 70.0 mm, young of year),

Lepomis macrochirus Ammonium

chloride 4 d F,M 8.75 18.5 12.7 227.4 Emery and Welch 1969

## 108

Appendix A. Acute Toxicity of Ammonia to Aquatic Animals

Species Chemical Name Duration Methods a pH Temp.

( ̊C) Total

Ammonia

(mg

TA N/L) Total Ammonia b

(mg TA N/L)

a djusted to pH 7

(all organisms )

and 20 ° C

( invertebrates ) SMAV

(mg

TA N/L) GMAV

(mg

TA N/L) Reference

Bluegill

(20.0

- 70.0 mm, young of year),

Lepomis macrochirus Ammonium

chloride 4 d F,M 9.05 18.5 6.581 193.8 Emery and Welch 1969

Bluegill

(20.0

- 70.0 mm, young of year),

Lepomis macrochirus Ammonium

chloride 4 d F,M 9.19 18.5 3.755 135.0 Emery and Welch 1969

Bluegill

(20.0

- 70.0 mm, young of year),

Lepomis macrochirus Ammonium

chloride 4 d F,M 9.62 18.5 0.7859 44.84 Emery and Welch 1969

Bluegill

(20.0

- 70.0 mm, young of year),

Lepomis macrochirus Ammonium

chloride 4 d F,M 9.85 18.5 1.346 89.70 Emery and Welch 1969

Bluegill,

Lepomis macrochirus Ammonium

chloride 4 d F,M 8.6 24 5.509 75.01 Hazel et al. 1979

Bluegill (5.2 cm),

Lepomis macrochirus Ammonium

chloride 4 d F,M 7.9 24.25 33.06 117.8 Lubinski et al. 1974

Bluegill (0.38 g, 26.3 mm),

Lepomis macrochirus Ammonium

chloride 4 d F,M 8.1 22 19.39 100.7 Mayes et al. 1986

Bluegill (19 mm, 0.0781 g),

Lepomis macrochirus Ammonium

chloride 4 d F,M 8.4 4 14.64 136.1 Reinbold and Pescitelli

1982b

Bluegill (22 mm, 0.1106 g),

Lepomis macrochirus Ammonium

chloride 4 d F,M 8.12 25 23.37 126.2 Reinbold and Pescitelli

1982b

Bluegill (28 mm, 0.250 g),

Lepomis macrochirus Ammonium

chloride 4 d F,M 8.16 4.5 12.55 73.19 Reinbold and Pescitelli

1982b

Bluegill (30 mm, 0.267 g),

Lepomis macrochirus Ammonium

chloride 4 d F,M 8.09 24.8 17.22 87.75 Reinbold and Pescitelli

1982b

Bluegill (217 mg),

Lepomis macrochirus Ammonium

chloride 4 d F,M 8 22 12.75 54.74 Roseboom and Richey 1977

Bluegill (342 mg),

Lepomis macrochirus Ammonium

chloride 4 d F,M 8.2 28 14.81 93.31 Roseboom and Richey 1977

Bluegill (646 mg),

Lepomis macrochirus Ammonium

chloride 4 d F,M 7.93 22 24.08 90.66 Roseboom and Richey 1977

Bluegill (72 mg),

Lepomis macrochirus Ammonium

chloride 4 d F,M 8.07 22 8.846 43.38 Roseboom and Richey 1977

Bluegill,

Lepomis macrochirus Ammonium

chloride 4 d F,M 7.6 21.7 44.03 93.31 Smith et al. 1984

## 109

Appendix A. Acute Toxicity of Ammonia to Aquatic Animals

Species Chemical Name Duration Methods a pH Temp.

( ̊C) Total

Ammonia

(mg

TA N/L) Total Ammonia b

(mg TA N/L)

a djusted to pH 7

(all organisms )

and 20 ° C

( invertebrates ) SMAV

(mg

TA N/L) GMAV

(mg

TA N/L) Reference

Bluegill (4.8 cm, 1.1 g),

Lepomis macrochirus Ammonium

chloride 4 d F,M 7.85 22.05 59.93 194.9 Sparks 1975

Bluegill (0.9 g),

Lepomis macrochirus Ammonium

chloride 4 d F,M 7.8 24.2 33.88 100.7 Swigert and Spacie 1983

Bluegill (0.9 g),

Lepomis macrochirus Ammonium

chloride 4 d F,M 7.6 26.5 58.69 124.4 Swigert and Spacie 1983

Bluegill (1.2 g),

Lepomis macrochirus Ammonium

chloride 4 d F,M 7.8 26.6 37.52 111.6 104.5 106.9 Swigert and Spacie 1983

Common carp (206 mg),

Cyprinus carpio Ammonium

chloride 4 d R,M 7.72 28 51.78 133.9 Hasan and MacIntosh 1986

Common carp (299 mg),

Cyprinus carpio Ammonium

chloride 4 d R,M 7.72 28 48.97 126.6 Hasan and MacIntosh 1986

Common carp (4

- 5 cm),

Cyprinus carpio Ammonium

chloride 4 d R,M 7.4 28 45.05 70.78 106.3 106.3 Rao et al. 1975

Golden trout (0.09 g, 24 cm),

Oncorhynchus aguabonita Ammonium

chloride 4 d F,M 8.06 13.2 23.3 112.1 112.1 Thurston and Russo 1981

Cutthroat trout (3.6 g),

Oncorhynchus clarkii Ammonium

chloride 4 d F,M 7.7 10 17.3 43.24 Thurston et al. 1981a

Cutthroat trout (3.6 g),

Oncorhynchus clarkii Ammonium

chloride 4 d F,M 7.7 10 29.1 72.73 Thurston et al. 1981a

Cutthroat trout (4.1 g),

Oncorhynchus clarkii Ammonium

chloride 4 d F,M 7.7 10 19.3 48.24 Thurston et al. 1981a

Cutthroat trout (4.1 g),

Oncorhynchus clarkii Ammonium

chloride 4 d F,M 7.7 10 26.3 65.73 Thurston et al. 1981a

Cutthroat trout (3.4 g),

Oncorhynchus clarkii Ammonium

chloride 4 d F,M 7.78 12.2 32.57 93.49 Thurston et al. 1978

Cutthroat trout (3.3 g),

Oncorhynchus clarkii Ammonium

chloride 4 d F,M 7.8 12.4 36.55 108.7 Thurston et al. 1978

Cutthroat trout (1.0 g),

Oncorhynchus clarkii Ammonium

chloride 4 d F,M 7.8 12.8 37.75 112.2 Thurston et al. 1978

Cutthroat trout (1.0 g),

Oncorhynchus clarkii Ammonium

chloride 4 d F,M 7.81 13.1 43.72 132.3 78.92 Thurston et al. 1978

## 110

Appendix A. Acute Toxicity of Ammonia to Aquatic Animals

Species Chemical Name Duration Methods a pH Temp.

( ̊C) Total

Ammonia

(mg

TA N/L) Total Ammonia b

(mg TA N/L)

a djusted to pH 7

(all organisms )

and 20 ° C

( invertebrates ) SMAV

(mg

TA N/L) GMAV

(mg

TA N/L) Reference

Pink salmon (late alevins),

Oncorhynchus gorbuscha Ammonium

sulfate 4 d S,M 6.4 4.3 230.5 164.6 Rice and Bailey 1980

Pink salmon (fry),

Oncorhynchus gorbuscha Ammonium

sulfate 4 d S,M 6.4 4.3 277.7 198.3 180.7 Rice and Bailey 1980

Coho salmon (6 g),

Oncorhynchus kisutch Ammonium

chloride 4 d F,M 8.1 17.2 11.59 60.20 Buckley 1978

Coho salmon,

Oncorhynchus kisutch Ammonium

chloride 4 d F,M 7 15 82.02 82.02 Wilson 1974; Robinson

Wilson and Seim 1975

Coho salmon,

Oncorhynchus kisutch Ammonium

chloride 4 d F,M 7 15 84.43 84.43 Wilson 1974; Robinson

Wilson and Seim 1975

Coho salmon,

Oncorhynchus kisutch Ammonium

chloride 4 d F,M 7.5 15 50.65 91.90 Wilson 1974; Robinson

Wilson and Seim 1975

Coho salmon,

Oncorhynchus kisutch Ammonium

chloride 4 d F,M 7.5 15 52.76 95.73 Wilson 1974; Robinson

Wilson and Seim 1975

Coho salmon,

Oncorhynchus kisutch Ammonium

chloride 4 d F,M 8 15 21.63 92.84 Wilson 1974; Robinson

Wilson and Seim 1975

Coho salmon,

Oncorhynchus kisutch Ammonium

chloride 4 d F,M 8 15 22 94.44 Wilson 1974; Robinson

Wilson and Seim 1975

Coho salmon,

Oncorhynchus kisutch Ammonium

chloride 4 d F,M 8.5 15 9.093 102.5 87.05 Wilson 1974; Robinson

Wilson and Seim 1975

Rainbow trout (0.5

- 3.0 g),

Oncorhynchus mykiss Ammonium

sulfate 4 d S,U 7.95 15 51.06 199.6 Qureshi et al. 1982

Rainbow trout

(McConaughy strain, 251 mg),

Oncorhynchus mykiss Ammonium

chloride 4 d S,M 6.84 12 112 98.86 Buhl and Hamilton 2000

Rainbow trout,

Oncorhynchus mykiss Ammonium

chloride 4 d S,M 7.55 15 34.23 67.04 Craig and Beggs 1979

Rainbow trout (0.80 g),

Oncorhynchus mykiss - 4 d S,M 6.95 14.7 163.6 156.9 Environment Canada 2004

Rainbow trout (0.60 g),

Oncorhynchus mykiss - 4 d S,M 6.97 14.5 144 140.3 Environment Canada 2004

Rainbow trout (0.63 g),

Oncorhynchus mykiss - 4 d S,M 7.02 15.4 146.7 149.4 Environment Canada 2004

## 111

Appendix A. Acute Toxicity of Ammonia to Aquatic Animals

Species Chemical Name Duration Methods a pH Temp.

( ̊C) Total

Ammonia

(mg

TA N/L) Total Ammonia b

(mg TA N/L)

a djusted to pH 7

(all organisms )

and 20 ° C

( invertebrates ) SMAV

(mg

TA N/L) GMAV

(mg

TA N/L) Reference

Rainbow trout (0.80 g),

Oncorhynchus mykiss - 4 d S,M 7.02 14.6 159 161.8 Environment Canada 2004

Rainbow trout (0.80 g),

Oncorhynchus mykiss - 4 d S,M 7.03 15.1 156.6 160.9 Environment Canada 2004

Rainbow trout (0.90 g),

Oncorhynchus mykiss - 4 d S,M 7.18 15.1 141.6 169.2 Environment Canada 2004

Rainbow trout (2.01 g),

Oncorhynchus mykiss - 4 d S,M 7.45 15.1 104.4 176.0 Environment Canada 2004

Rainbow trout (1.30 g),

Oncorhynchus mykiss - 4 d S,M 7.47 14.7 72.65 126.1 Environment Canada 2004

Rainbow trout (0.78 g),

Oncorhynchus mykiss - 4 d S,M 7.47 14.5 79.67 138.3 Environment Canada 2004

Rainbow trout (0.40 g),

Oncorhynchus mykiss - 4 d S,M 7.51 14.2 73.71 135.8 Environment Canada 2004

Rainbow trout (1.64 g),

Oncorhynchus mykiss - 4 d S,M 7.54 14.6 75.3 145.2 Environment Canada 2004

Rainbow trout (1.13 g),

Oncorhynchus mykiss - 4 d S,M 7.59 13.9 59.4 123.9 Environment Canada 2004

Rainbow trout (1.50 g),

Oncorhynchus mykiss - 4 d S,M 7.87 15.1 42.9 144.7 Environment Canada 2004

Rainbow trout (1.38 g),

Oncorhynchus mykiss - 4 d S,M 7.93 15.2 41.15 155.0 Environment Canada 2004

Rainbow trout (0.90 g),

Oncorhynchus mykiss - 4 d S,M 7.97 15.2 36.17 145.4 Environment Canada 2004

Rainbow trout (1.00 g),

Oncorhynchus mykiss - 4 d S,M 7.98 15.1 35.29 145.9 Environment Canada 2004

Rainbow trout (1.30 g),

Oncorhynchus mykiss - 4 d S,M 8.03 14.9 23.03 104.6 Environment Canada 2004

Rainbow trout (1.26 g),

Oncorhynchus mykiss - 4 d S,M 8.04 14.3 25.84 119.7 Environment Canada 2004

Rainbow trout (1.60 g),

Oncorhynchus mykiss - 4 d S,M 8.34 15.3 19.15 158.5 Environment Canada 2004

Rainbow trout (1.30 g),

Oncorhynchus mykiss - 4 d S,M 8.39 15.3 12.05 109.9 Environment Canada 2004

Rainbow trout (1.11 g),

Oncorhynchus mykiss - 4 d S,M 8.4 14.9 12.84 119.4 Environment Canada 2004

## 112

Appendix A. Acute Toxicity of Ammonia to Aquatic Animals

Species Chemical Name Duration Methods a pH Temp.

( ̊C) Total

Ammonia

(mg

TA N/L) Total Ammonia b

(mg TA N/L)

a djusted to pH 7

(all organisms )

and 20 ° C

( invertebrates ) SMAV

(mg

TA N/L) GMAV

(mg

TA N/L) Reference

Rainbow trout (1.40 g),

Oncorhynchus mykiss - 4 d S,M 8.44 14.7 14.41 144.7 Environment Canada 2004

Rainbow trout (0.90 g),

Oncorhynchus mykiss - 4 d S,M 8.46 14.5 11.82 123.4 Environment Canada 2004

Rainbow trout (1.26 g),

Oncorhynchus mykiss - 4 d S,M 8.47 14.3 17.2 183.0 Environment Canada 2004

Rainbow trout (1.01 g),

Oncorhynchus mykiss - 4 d S,M 8.93 14.2 4.8 117.0 Environment Canada 2004

Rainbow trout (1.44 g),

Oncorhynchus mykiss - 4 d S,M 8.93 15 5.4 131.6 Environment Canada 2004

Rainbow trout (1.42 g),

Oncorhynchus mykiss - 4 d S,M 9.46 14.6 1.6 79.03 Environment Canada 2004

Rainbow trout,

Oncorhynchus mykiss - 4 d S,M 7.5 15 38.37 69.63 Holt and Malcolm 1979

Rainbow trout (129 mm),

Oncorhynchus mykiss Phosphoric acid,

Diammonium

salt 4 d F,U 7 15 207.5 207.5 Blahm 1978

Rainbow trout (1.7

- 1.9 cm),

Oncorhynchus mykiss Ammonium

chloride 4 d F,U 7.4 14.5 20.03 31.47 Calamari et al. 1981

Rainbow trout,

Oncorhynchus mykiss Ammonium

chloride 4 d F,U 7.4 14.5 46.31 72.77 Calamari et al. 1981

Rainbow trout (8

- 10 cm),

Oncorhynchus mykiss Ammonium

chloride 4 d F,U 7.4 14.5 55.07 86.53 Calamari et al. 1981

Rainbow trout (129 mm),

Oncorhynchus mykiss Phosphoric acid,

Diammonium

salt 4 d F,U 8 15 70 300.5 Blahm 1978

Rainbow trout (10.9 g),

Oncorhynchus mykiss Ammonium

chloride 4 d F,M 7.7 3.6 38.52 96.27 West 1985; Arthur et al.

1987

Rainbow trout (14.0 g),

Oncorhynchus mykiss Ammonium

chloride 4 d F,M 7.7 9.8 55.15 137.8 West 1985; Arthur et al.

1987

Rainbow trout (22.4 g),

Oncorhynchus mykiss Ammonium

chloride 4 d F,M 7.9 16.2 15.23 54.24 West 1985; Arthur et al.

1987

Rainbow trout (10.3 g),

Oncorhynchus mykiss Ammonium

chloride 4 d F,M 7.9 11.3 30.15 107.4 West 1985; Arthur et al.

1987

Rainbow trout (3.3 g),

Oncorhynchus mykiss Ammonium

chloride 4 d F,M 8.3 18.7 12.75 97.57 West 1985; Arthur et al.

1987

## 113

Appendix A. Acute Toxicity of Ammonia to Aquatic Animals

Species Chemical Name Duration Methods a pH Temp.

( ̊C) Total

Ammonia

(mg

TA N/L) Total Ammonia b

(mg TA N/L)

a djusted to pH 7

(all organisms )

and 20 ° C

( invertebrates ) SMAV

(mg

TA N/L) GMAV

(mg

TA N/L) Reference

Rainbow trout (53 mm, 1.48 g),

Oncorhynchus mykiss Ammonium

chloride 4 d F,M 7.95 10 35.14 137.3 Broderius and Smith Jr.

1979

Rainbow trout (stage 8),

Oncorhynchus mykiss Ammonium

chloride 4 d F,M 7.4 14.4 40.99 64.40 Calamari et al. 1977

Rainbow trout,

Oncorhynchus mykiss Ammonium

chloride 4 d F,M 8.05 14 22.9 108.1 DeGraeve et al. 1980

Rainbow trout (45 mm, 0.86 g),

Oncorhynchus mykiss Ammonium

chloride 4 d F,M 8.16 14.2 23.39 136.4 Reinbold and Pescitelli

1982b

Rainbow trout

(119 mm, 20.6 g),

Oncorhynchus mykiss Ammonium

chloride 4 d F,M 8.28 12.8 15.4 113.4 Reinbold and Pescitelli

1982b

Rainbow trout

(115 mm, 18.1 g),

Oncorhynchus mykiss Ammonium

chloride 4 d F,M 8.34 5 17.32 143.3 Reinbold and Pescitelli

1982b

Rainbow trout (42 mm, 0.61 g),

Oncorhynchus mykiss Ammonium

chloride 4 d F,M 8.43 3 11.86 116.8 Reinbold and Pescitelli

1982b

Rainbow trout (52 mm, 1.47 g),

Oncorhynchus mykiss Ammonium

chloride 4 d F,M 8.5 14.9 10.09 113.7 Reinbold and Pescitelli

1982b

Rainbow trout (44 mm, 0.76 g),

Oncorhynchus mykiss Ammonium

chloride 4 d F,M 8.6 3.3 15.27 207.9 Reinbold and Pescitelli

1982b

Rainbow trout (6.3 g, 8.1 cm),

Oncorhynchus mykiss Ammonium

chloride 4 d F,M 7.44 12.8 32.49 54.00 Thurston and Russo 1983

Rainbow trout (8.0 g, 8.9 cm),

Oncorhynchus mykiss Ammonium

chloride 4 d F,M 7.5 14.5 24.2 43.91 Thurston and Russo 1983

Rainbow trout

(29.8 g, 13.1 cm),

Oncorhynchus mykiss Ammonium

chloride 4 d F,M 7.59 12.7 32.62 68.03 Thurston and Russo 1983

Rainbow trout

(28.0 g, 13.1 cm),

Oncorhynchus mykiss Ammonium

chloride 4 d F,M 7.6 13 23.8 50.43 Thurston and Russo 1983

Rainbow trout

(24.5 g, 12.7 cm),

Oncorhynchus mykiss Ammonium

chloride 4 d F,M 7.6 12.9 25.14 53.27 Thurston and Russo 1983

Rainbow trout

(2596 g, 57.0 cm),

Oncorhynchus mykiss Ammonium

chloride 4 d F,M 7.62 7.9 20.53 44.93 f Thurston and Russo 1983

## 114

Appendix A. Acute Toxicity of Ammonia to Aquatic Animals

Species Chemical Name Duration Methods a pH Temp.

( ̊C) Total

Ammonia

(mg

TA N/L) Total Ammonia b

(mg TA N/L)

a djusted to pH 7

(all organisms )

and 20 ° C

( invertebrates ) SMAV

(mg

TA N/L) GMAV

(mg

TA N/L) Reference

Rainbow trout

(15.1 g, 10.7 cm),

Oncorhynchus mykiss Ammonium

chloride 4 d F,M 7.62 14.4 28.62 62.64 Thurston and Russo 1983

Rainbow trout

(29.6 g, 13.3 cm),

Oncorhynchus mykiss Ammonium

chloride 4 d F,M 7.63 12.9 25.65 57.06 Thurston and Russo 1983

Rainbow trout

(1496 g, 48.5 cm),

Oncorhynchus mykiss Ammonium

chloride 4 d F,M 7.64 9.8 25.82 58.38 f Thurston and Russo 1983

Rainbow trout

(18.9 g, 11.6 cm),

Oncorhynchus mykiss Ammonium

chloride 4 d F,M 7.64 13.1 29.28 66.21 Thurston and Russo 1983

Rainbow trout

(558 g, 37.0 cm),

Oncorhynchus mykiss Ammonium

chloride 4 d F,M 7.64 10 31.85 72.02 Thurston and Russo 1983

Rainbow trout

(1698 g, 50.9 cm),

Oncorhynchus mykiss Ammonium

chloride 4 d F,M 7.65 9.8 19.46 44.73 f Thurston and Russo 1983

Rainbow trout

(22.8 g, 12.3 cm),

Oncorhynchus mykiss Ammonium

chloride 4 d F,M 7.65 13.2 28.64 65.84 Thurston and Russo 1983

Rainbow trout

(12.3 g, 10.2 cm),

Oncorhynchus mykiss Ammonium

chloride 4 d F,M 7.65 14.3 29.02 66.71 Thurston and Russo 1983

Rainbow trout

(513 g, 35.9 cm),

Oncorhynchus mykiss Ammonium

chloride 4 d F,M 7.66 9.8 25.95 60.65 Thurston and Russo 1983

Rainbow trout

(22.6 g, 12.3 cm),

Oncorhynchus mykiss Ammonium

chloride 4 d F,M 7.66 13.6 28.27 66.07 Thurston and Russo 1983

Rainbow trout

(26.0 g, 13.0 cm),

Oncorhynchus mykiss Ammonium

chloride 4 d F,M 7.66 12.8 33.97 79.39 Thurston and Russo 1983

Rainbow trout

(14.8 g, 10.5 cm),

Oncorhynchus mykiss Ammonium

chloride 4 d F,M 7.67 14 27.3 64.87 Thurston and Russo 1983

## 115

Appendix A. Acute Toxicity of Ammonia to Aquatic Animals

Species Chemical Name Duration Methods a pH Temp.

( ̊C) Total

Ammonia

(mg

TA N/L) Total Ammonia b

(mg TA N/L)

a djusted to pH 7

(all organisms )

and 20 ° C

( invertebrates ) SMAV

(mg

TA N/L) GMAV

(mg

TA N/L) Reference

Rainbow trout

(38.0 g, 14.4 cm),

Oncorhynchus mykiss Ammonium

chloride 4 d F,M 7.68 13 33.15 80.11 Thurston and Russo 1983

Rainbow trout

(1122 g, 45.6 cm),

Oncorhynchus mykiss Ammonium

chloride 4 d F,M 7.69 10.4 17.75 43.62 f Thurston and Russo 1983

Rainbow trout

(1140 g, 46.4 cm),

Oncorhynchus mykiss Ammonium

chloride 4 d F,M 7.69 10.7 20.18 49.59 f Thurston and Russo 1983

Rainbow trout

(152 g, 23.4 cm),

Oncorhynchus mykiss Ammonium

chloride 4 d F,M 7.69 10.7 25.62 62.96 Thurston and Russo 1983

Rainbow trout

(23.6 g, 13.2 cm),

Oncorhynchus mykiss Ammonium

chloride 4 d F,M 7.69 13.4 27.51 67.60 Thurston and Russo 1983

Rainbow trout (9.5 g, 9.4 cm),

Oncorhynchus mykiss Ammonium

chloride 4 d F,M 7.9 12.7 20.03 71.36 Thurston and Russo 1983

Rainbow trout (4.3 g, 7.1 cm),

Oncorhynchus mykiss Ammonium

chloride 4 d F,M 7.71 11.5 30.22 76.83 Thurston and Russo 1983

Rainbow trout (4.0 g, 7.0 cm),

Oncorhynchus mykiss Ammonium

chloride 4 d F,M 7.71 11.4 32.02 81.40 Thurston and Russo 1983

Rainbow trout

(248 g, 25.2 cm),

Oncorhynchus mykiss Ammonium

chloride 4 d F,M 7.74 10.4 25.76 68.95 Thurston and Russo 1983

Rainbow trout

(25.8 g, 13.6 cm),

Oncorhynchus mykiss Ammonium

chloride 4 d F,M 7.75 11.8 31.53 85.87 Thurston and Russo 1983

Rainbow trout (8.1 g, 9.3 cm),

Oncorhynchus mykiss Ammonium

chloride 4 d F,M 7.75 12.3 33.94 92.43 Thurston and Russo 1983

Rainbow trout

(380 g, 32.4 cm),

Oncorhynchus mykiss Ammonium

chloride 4 d F,M 7.76 10 22.44 62.19 Thurston and Russo 1983

Rainbow trout

(42.0 g, 16.0 cm),

Oncorhynchus mykiss Ammonium

chloride 4 d F,M 7.77 13.6 31.81 89.71 Thurston and Russo 1983

## 116

Appendix A. Acute Toxicity of Ammonia to Aquatic Animals

Species Chemical Name Duration Methods a pH Temp.

( ̊C) Total

Ammonia

(mg

TA N/L) Total Ammonia b

(mg TA N/L)

a djusted to pH 7

(all organisms )

and 20 ° C

( invertebrates ) SMAV

(mg

TA N/L) GMAV

(mg

TA N/L) Reference

Rainbow trout (1.7 g, 5.7 cm),

Oncorhynchus mykiss Ammonium

chloride 4 d F,M 7.79 12.4 41.97 122.6 Thurston and Russo 1983

Rainbow trout

(11.2 g, 10.0 cm),

Oncorhynchus mykiss Ammonium

chloride 4 d F,M 7.8 9.7 23.65 70.32 Thurston and Russo 1983

Rainbow trout (5.7 g, 8.4 cm),

Oncorhynchus mykiss Ammonium

chloride 4 d F,M 7.8 13.3 42.02 124.9 Thurston and Russo 1983

Rainbow trout (2.3 g, 6.1 cm),

Oncorhynchus mykiss Ammonium

chloride 4 d F,M 7.8 12.4 47.87 142.3 Thurston and Russo 1983

Rainbow trout (8.0 g, 9.5 cm),

Oncorhynchus mykiss Ammonium

chloride 4 d F,M 7.82 13.2 33.67 103.7 Thurston and Russo 1983

Rainbow trout (4.6 g, 7.4 cm),

Oncorhynchus mykiss Ammonium

chloride 4 d F,M 7.83 13.5 33.55 105.2 Thurston and Russo 1983

Rainbow trout (6.7 g, 8.6 cm),

Oncorhynchus mykiss Ammonium

chloride 4 d F,M 7.84 12.2 24.54 78.38 Thurston and Russo 1983

Rainbow trout (9.0 g, 9.3 cm),

Oncorhynchus mykiss Ammonium

chloride 4 d F,M 7.84 12.9 32.3 103.2 Thurston and Russo 1983

Rainbow trout (1.8 g, 5.7 cm),

Oncorhynchus mykiss Ammonium

chloride 4 d F,M 7.84 13.8 33.09 105.7 Thurston and Russo 1983

Rainbow trout (4.3 g, 7.1 cm),

Oncorhynchus mykiss Ammonium

chloride 4 d F,M 7.84 13 38.69 123.6 Thurston and Russo 1983

Rainbow trout (0.47 g, 4.0 cm),

Oncorhynchus mykiss Ammonium

chloride 4 d F,M 7.85 12.5 29.77 96.81 Thurston and Russo 1983

Rainbow trout (2.5 g, 6.0 cm),

Oncorhynchus mykiss Ammonium

chloride 4 d F,M 7.85 13.1 31.55 102.6 Thurston and Russo 1983

Rainbow trout (0.61 g, 4.3 cm),

Oncorhynchus mykiss Ammonium

chloride 4 d F,M 7.85 13.1 33.59 109.2 Thurston and Russo 1983

Rainbow trout (1.02 g, 4.9 cm),

Oncorhynchus mykiss Ammonium

chloride 4 d F,M 7.85 12.3 33.99 110.5 Thurston and Russo 1983

Rainbow trout (9.4 g, 9.6 cm),

Oncorhynchus mykiss Ammonium

chloride 4 d F,M 7.85 16.1 34.17 111.1 Thurston and Russo 1983

Rainbow trout (0.33 g, 3.6 cm),

Oncorhynchus mykiss Ammonium

chloride 4 d F,M 7.86 13 20.7 68.55 Thurston and Russo 1983

Rainbow trout (0.33 g, 3.6 cm),

Oncorhynchus mykiss Ammonium

chloride 4 d F,M 7.86 13.4 23.71 78.52 Thurston and Russo 1983

## 117

Appendix A. Acute Toxicity of Ammonia to Aquatic Animals

Species Chemical Name Duration Methods a pH Temp.

( ̊C) Total

Ammonia

(mg

TA N/L) Total Ammonia b

(mg TA N/L)

a djusted to pH 7

(all organisms )

and 20 ° C

( invertebrates ) SMAV

(mg

TA N/L) GMAV

(mg

TA N/L) Reference

Rainbow trout (0.47 g, 4.0 cm),

Oncorhynchus mykiss Ammonium

chloride 4 d F,M 7.86 12.7 28.77 95.27 Thurston and Russo 1983

Rainbow trout (1.7 g, 5.8 cm),

Oncorhynchus mykiss Ammonium

chloride 4 d F,M 7.86 14.1 34.95 115.7 Thurston and Russo 1983

Rainbow trout

(48.6 g, 15.2 cm),

Oncorhynchus mykiss Ammonium

chloride 4 d F,M 7.86 10.2 35.31 116.9 Thurston and Russo 1983

Rainbow trout (0.15 g, 2.7 cm),

Oncorhynchus mykiss Ammonium

chloride 4 d F,M 7.87 12.9 16.81 56.69 Thurston and Russo 1983

Rainbow trout (0.18 g, 2.9 cm),

Oncorhynchus mykiss Ammonium

chloride 4 d F,M 7.87 12.9 18.99 64.04 Thurston and Russo 1983

Rainbow trout (0.23 g, 3.2 cm),

Oncorhynchus mykiss Ammonium

chloride 4 d F,M 7.87 13.1 19.08 64.34 Thurston and Russo 1983

Rainbow trout (7.0 g, 8.8 cm),

Oncorhynchus mykiss Ammonium

chloride 4 d F,M 7.87 12.2 20.02 67.51 Thurston and Russo 1983

Rainbow trout (0.18 g, 2.9 cm),

Oncorhynchus mykiss Ammonium

chloride 4 d F,M 7.87 13 21.15 71.32 Thurston and Russo 1983

Rainbow trout (2.6 g, 6.2 cm),

Oncorhynchus mykiss Ammonium

chloride 4 d F,M 7.87 12.1 31.8 107.2 Thurston and Russo 1983

Rainbow trout (11.1 g, 9.9 cm),

Oncorhynchus mykiss Ammonium

chloride 4 d F,M 7.87 13 34.32 115.7 Thurston and Russo 1983

Rainbow trout (0.12 g, 2.4 cm),

Oncorhynchus mykiss Ammonium

chloride 4 d F,M 7.88 12.8 11.07 38.02 Thurston and Russo 1983

Rainbow trout (0.14 g, 2.6 cm),

Oncorhynchus mykiss Ammonium

chloride 4 d F,M 7.88 12.9 15.91 54.64 Thurston and Russo 1983

Rainbow trout (0.23 g, 3.2 cm),

Oncorhynchus mykiss Ammonium

chloride 4 d F,M 7.88 13.4 19.43 66.73 Thurston and Russo 1983

Rainbow trout

(52.1 g, 15.5 cm),

Oncorhynchus mykiss Ammonium

chloride 4 d F,M 7.88 10 28.6 98.22 Thurston and Russo 1983

Rainbow trout (1.8 g, 5.9 cm),

Oncorhynchus mykiss Ammonium

sulfate 4 d F,M 7.89 12.4 36.73 128.5 Thurston and Russo 1983

Rainbow trout (0.06 g, 1.7 cm),

Oncorhynchus mykiss Ammonium

chloride 4 d F,M 7.9 13.4 19.44 69.26 Thurston and Russo 1983

Rainbow trout (0.06 g, 1.7 cm),

Oncorhynchus mykiss Ammonium

chloride 4 d F,M 7.7 13.9 28.54 71.33 Thurston and Russo 1983

## 118

Appendix A. Acute Toxicity of Ammonia to Aquatic Animals

Species Chemical Name Duration Methods a pH Temp.

( ̊C) Total

Ammonia

(mg

TA N/L) Total Ammonia b

(mg TA N/L)

a djusted to pH 7

(all organisms )

and 20 ° C

( invertebrates ) SMAV

(mg

TA N/L) GMAV

(mg

TA N/L) Reference

Rainbow trout (7.9 g, 9.2 cm),

Oncorhynchus mykiss Ammonium

chloride 4 d F,M 7.9 11.9 22.65 80.69 Thurston and Russo 1983

Rainbow trout (9.7 g, 9.7 cm),

Oncorhynchus mykiss Ammonium

chloride 4 d F,M 7.9 13 35.75 127.4 Thurston and Russo 1983

Rainbow trout (9.3 g, 9.0 cm),

Oncorhynchus mykiss Ammonium

chloride 4 d F,M 7.9 13 37.41 133.3 Thurston and Russo 1983

Rainbow trout (0.08 g, 2.0 cm),

Oncorhynchus mykiss Ammonium

chloride 4 d F,M 7.91 13.1 12.68 46.01 Thurston and Russo 1983

Rainbow trout (0.06 g, 1.7 cm),

Oncorhynchus mykiss Ammonium

chloride 4 d F,M 7.91 13 20.99 76.17 Thurston and Russo 1983

Rainbow trout (7.1 g, 8.4 cm),

Oncorhynchus mykiss Ammonium

chloride 4 d F,M 7.91 19 25.36 92.03 Thurston and Russo 1983

Rainbow trout (10.1 g, 9.8 cm),

Oncorhynchus mykiss Ammonium

chloride 4 d F,M 7.91 19.1 26.44 95.95 Thurston and Russo 1983

Rainbow trout (1.7 g, 5.8 cm),

Oncorhynchus mykiss Phosphoric acid,

Diammonium

salt 4 d F,M 7.94 12.8 26.49 101.6 Thurston and Russo 1983

Rainbow trout (2.1 g, 6.2 cm),

Oncorhynchus mykiss Ammonium

sulfate 4 d F,M 7.94 12.5 39.25 150.6 Thurston and Russo 1983

Rainbow trout (0.15 g, 2.7 cm),

Oncorhynchus mykiss Ammonium

chloride 4 d F,M 7.95 12.5 19.75 77.19 Thurston and Russo 1983

Rainbow trout (8.6 g, 8.9 cm),

Oncorhynchus mykiss Ammonium

chloride 4 d F,M 7.96 19.2 23.21 92.42 Thurston and Russo 1983

Rainbow trout (2.1 g, 6.2 cm),

Oncorhynchus mykiss Phosphoric acid,

Diammonium

salt 4 d F,M 7.98 12.5 27.02 111.7 Thurston and Russo 1983

Rainbow trout (1.01 g, 4.6 cm),

Oncorhynchus mykiss Ammonium

chloride 4 d F,M 8.06 13.2 33.64 161.8 Thurston and Russo 1983

Rainbow trout (0.36 g, 3.4 cm),

Oncorhynchus mykiss Ammonium

chloride 4 d F,M 8.08 12.8 23.05 115.2 Thurston and Russo 1983

Rainbow trout (1.7 g, 5.9 cm),

Oncorhynchus mykiss Ammonium

bicarbonate 4 d F,M 8.1 13.9 18.14 94.23 Thurston and Russo 1983

Rainbow trout (1.8 g, 5.8 cm),

Oncorhynchus mykiss Ammonium

bicarbonate 4 d F,M 8.12 13.6 17.34 93.61 Thurston and Russo 1983

Rainbow trout (2596 g),

Oncorhynchus mykiss Ammonium

chloride 4 d F,M 7.62 7.9 21.6 47.27 Thurston et al. 1981a

## 119

Appendix A. Acute Toxicity of Ammonia to Aquatic Animals

Species Chemical Name Duration Methods a pH Temp.

( ̊C) Total

Ammonia

(mg

TA N/L) Total Ammonia b

(mg TA N/L)

a djusted to pH 7

(all organisms )

and 20 ° C

( invertebrates ) SMAV

(mg

TA N/L) GMAV

(mg

TA N/L) Reference

Rainbow trout (2080 g),

Oncorhynchus mykiss Ammonium

chloride 4 d F,M 7.67 7.7 17 40.40 Thurston et al. 1981a

Rainbow trout (293 g),

Oncorhynchus mykiss Ammonium

chloride 4 d F,M 7.71 8.5 20.7 52.62 Thurston et al. 1981a

Rainbow trout (230 g),

Oncorhynchus mykiss Ammonium

chloride 4 d F,M 7.72 8.2 10.5 27.15 Thurston et al. 1981a

Rainbow trout (244 g),

Oncorhynchus mykiss Ammonium

chloride 4 d F,M 7.72 8.1 19.8 51.20 Thurston et al. 1981a

Rainbow trout (230 g),

Oncorhynchus mykiss Ammonium

chloride 4 d F,M 7.74 8.3 22.3 59.69 Thurston et al. 1981a

Rainbow trout (247 g),

Oncorhynchus mykiss Ammonium

chloride 4 d F,M 7.74 8.1 28 74.94 Thurston et al. 1981a

Rainbow trout (18 g),

Oncorhynchus mykiss Ammonium

chloride 4 d F,M 7.86 9.6 19.3 63.91 Thurston et al. 1981a

Rainbow trout (21 g),

Oncorhynchus mykiss Ammonium

chloride 4 d F,M 7.86 9.7 31.6 104.6 Thurston et al. 1981a

Rainbow trout (4.6 g, 7.3 cm),

Oncorhynchus mykiss Ammonium

chloride 4 d F,M 7.75 12.7 32.09 87.39 Thurston et al. 1981b

Rainbow trout (5.7 g, 8.0 cm),

Oncorhynchus mykiss Ammonium

chloride 4 d F,M 7.75 12.5 36.97 100.7 Thurston et al. 1981b

Rainbow trout (5.0 g, 7.6 cm),

Oncorhynchus mykiss Ammonium

chloride 4 d F,M 7.76 12.5 39.08 108.3 Thurston et al. 1981b

Rainbow trout (5.7 g, 8.0 cm),

Oncorhynchus mykiss Ammonium

chloride 4 d F,M 7.79 12.9 40.88 119.4 Thurston et al. 1981b

Rainbow trout (4.0 g, 7.2 cm),

Oncorhynchus mykiss Ammonium

chloride 4 d F,M 7.83 12.8 36.49 114.5 Thurston et al. 1981b

Rainbow trout (9.5 g, 9.4 cm),

Oncorhynchus mykiss Ammonium

chloride 4 d F,M 6.51 14.1 157.4 116.8 Thurston et al. 1981c

Rainbow trout (9.5 g, 9.4 cm),

Oncorhynchus mykiss Ammonium

chloride 4 d F,M 6.8 14.1 94.05 80.83 Thurston et al. 1981c

Rainbow trout (9.5 g, 9.4 cm),

Oncorhynchus mykiss Ammonium

chloride 4 d F,M 7.3 14 74.2 102.2 Thurston et al. 1981c

Rainbow trout (9.5 g, 9.4 cm),

Oncorhynchus mykiss Ammonium

chloride 4 d F,M 8.29 14.1 13.85 104.0 Thurston et al. 1981c

Rainbow trout (9.5 g, 9.4 cm),

Oncorhynchus mykiss Ammonium

chloride 4 d F,M 8.82 13.9 3.95 80.02 Thurston et al. 1981c

## 120

Appendix A. Acute Toxicity of Ammonia to Aquatic Animals

Species Chemical Name Duration Methods a pH Temp.

( ̊C) Total

Ammonia

(mg

TA N/L) Total Ammonia b

(mg TA N/L)

a djusted to pH 7

(all organisms )

and 20 ° C

( invertebrates ) SMAV

(mg

TA N/L) GMAV

(mg

TA N/L) Reference

Rainbow trout (9.5 g, 9.4 cm),

Oncorhynchus mykiss Ammonium

chloride 4 d F,M 9.01 14.5 2.51 69.50 Thurston et al. 1981c

Rainbow trout (juvenile),

Oncorhynchus mykiss Ammonium

chloride 4 d F,M 7.2 10 174 212.6 Wicks and Randall 2002

Rainbow trout

(40.0 g; swimming fish),

Oncorhynchus mykiss Ammonium

chloride 4 d F,M 6.97 16.6 32.38 31.56 Wicks et al. 2002

Rainbow trout

(40.0 g; resting fish),

Oncorhynchus mykiss Ammonium

chloride 4 d F,M 6.97 16.6 207 201.7 82.88 Wicks et al. 2002

Chinook salmon (1.0

- 7 g),

Oncorhynchus tshawytscha Ammonia 4 d S,M 7.96 7 28.03 111.6 Servizi and Gordon 1990

Chinook salmon

(14.4 g, 11.9 cm),

Oncorhynchus tshawytscha Ammonium

chloride 4 d F,U 7.87 13.5 18.47 62.29 Thurston and Meyn 1984

Chinook salmon

(15.3 g, 12.1 cm),

Oncorhynchus tshawytscha Ammonium

chloride 4 d F,U 7.82 12.2 27.23 83.90 Thurston and Meyn 1984

Chinook salmon

(18.1 g, 12.7 cm),

Oncorhynchus tshawytscha Ammonium

chloride 4 d F,U 7.84 12.3 24.74 79.02 82.39 99.15 Thurston and Meyn 1984

Topeka shiner (adult, 29 mo),

Notropis topeka Ammonium

chloride 4 d F,M 7.85 24.6 21.40 69.59 Adelman et al. 2009

Topeka shiner

(juvenile, 16 mo),

Notropis topeka Ammonium

chloride 4 d F,M 8.05 25.0 18.70 88.27 Adelman et al. 2009

Topeka shiner

(juvenile, 15 mo),

Notropis topeka Ammonium

chloride 4 d F,M 8.09 13.2 28.90 147.3 96.72 96.72 Adelman et al. 2009

Leopard frog (embryo),

Rana pipiens Ammonium

chloride 4 d F,M 8 20 31.04 133.3 Diamond et al. 1993

Leopard frog (8 d),

Rana pipiens Ammonium

chloride 4 d F,M 8 12 16.23 69.69 96.38 96.38 Diamond et al. 1993

## 121

Appendix A. Acute Toxicity of Ammonia to Aquatic Animals

Species Chemical Name Duration Methods a pH Temp.

( ̊C) Total

Ammonia

(mg

TA N/L) Total Ammonia b

(mg TA N/L)

a djusted to pH 7

(all organisms )

and 20 ° C

( invertebrates ) SMAV

(mg

TA N/L) GMAV

(mg

TA N/L) Reference

Long fingernailclam,

Musculium transversum Ammonium

chloride 4 d F,M 8.1 14.6 32.83 109.0 West 1985; Arthur et al.

1987

Long fingernailclam,

Musculium transversum Ammonium

chloride 4 d F,M 8.2 5.4 38.18 71.74 West 1985; Arthur et al.

1987

Long fingernailclam,

Musculium transversum Ammonium

chloride 4 d F,M 8.6 20.5 6.429 91.24 89.36 89.36 West 1985; Arthur et al.

1987

Smallmouth bass

(26

- 29 mm, 264

- 267 mg),

Micropterus dolomieu Ammonium

chloride 4 d F,M 7.16 22.3 123.4 144.3 Broderius et al. 1985

Smallmouth bass

(26

- 29 mm, 264

- 267 mg),

Micropterus dolomieu Ammonium

chloride 4 d F,M 6.53 22.3 359.9 269.2 Broderius et al. 1985

Smallmouth bass

(26

- 29 mm, 264

- 267 mg),

Micropterus dolomieu Ammonium

chloride 4 d F,M 7.74 22.3 39.3 105.2 Broderius et al. 1985

Smallmouth bass

(26

- 29 mm, 264

- 267 mg),

Micropterus dolomieu Ammonium

chloride 4 d F,M 8.71 22.3 7.56 126.0 150.6 Broderius et al. 1985

Largemouth bass

(0.086

- 0.322 g),

Micropterus salmoides Ammonium

chloride 4 d F,M 8.04 28 19.59 90.72 Roseboom and Richey 1977

Largemouth bass

(2.018

- 6.286 g),

Micropterus salmoides Ammonium

chloride 4 d F,M 7.96 22 20.48 81.56 86.02 Roseboom and Richey 1977

Guadalupe bass (6.5 g),

Micropterus treculii Ammonium

chloride 4 d S,M/ 8 22 12.7 54.52 54.52 89.06 Tomasso and Carmichael

1986

Great pond snail (25

- 30 mm),

Lymnaea stagnalis - 4 d F,M 7.9 11.5 50.33 88.62 88.62 88.62 Williams et al. 1986

Guppy (0.13 g, 2.03 cm),

Poecilia reticulata Ammonium

chloride 4 d S,U 7.5 27.55 5.929 10.76 Kumar and Krishnamoorthi

1983

## 122

Appendix A. Acute Toxicity of Ammonia to Aquatic Animals

Species Chemical Name Duration Methods a pH Temp.

( ̊C) Total

Ammonia

(mg

TA N/L) Total Ammonia b

(mg TA N/L)

a djusted to pH 7

(all organisms )

and 20 ° C

( invertebrates ) SMAV

(mg

TA N/L) GMAV

(mg

TA N/L) Reference

Guppy (8.0 mm),

Poecilia reticulata - 4 d S,U 7.22 25 129.4 161.8 Rubin and Elmaraghy 1976

Guppy (8.25(6.3

- 11.0) mm),

Poecilia reticulata - 4 d S,U 7.45 25 75.65 127.6 Rubin and Elmaraghy 1976

Guppy (8.70(6.8

- 10.6) mm),

Poecilia reticulata - 4 d S,U 7.45 25 82.95 139.9 74.66 74.66 Rubin and Elmaraghy 1976

Johnny darter (38 mm),

Etheostoma nigrum Ammonium

chloride 4 d F,M 7.9 20.6 28.9 103.0 Nimmo et al. 1989

Johnny darter (38 mm),

Etheostoma nigrum Ammonium

chloride 4 d F,M 8 20.1 24.61 105.7 Nimmo et al. 1989

Johnny darter (38 mm),

Etheostoma nigrum Ammonium

chloride 4 d F,M 8.2 6.2 6.937 43.72 Nimmo et al. 1989

Johnny darter (38 mm),

Etheostoma nigrum Ammonium

chloride 4 d F,M 8.1 5.8 11.47 59.57 Nimmo et al. 1989

Johnny darter (38 mm),

Etheostoma nigrum Ammonium

chloride 4 d F,M 8.1 5.8 13.46 69.93 Nimmo et al. 1989

Johnny darter (38 mm),

Etheostoma nigrum Ammonium

chloride 4 d F,M 8 20.1 15.63 67.08 71.45 Nimmo et al. 1989

Orangethroat darter,

Etheostoma spectabile Ammonium

chloride 4 d F,M 8.1 22 16.12 83.74 Hazel et al. 1979

Orangethroat darter,

Etheostoma spectabile Ammonium

chloride 4 d F,M 8.4 21 7.65 71.12 77.17 74.25 Hazel et al. 1979

Rio Grande silvery minnow

(3

- 5 d old),

Hybognathus amarus Ammonium

chloride 4 d R,M 8 25 16.9 72.55 72.55 72.55 Buhl 2002

Spring peeper (embryo),

Pseudacris crucifer Ammonium

chloride 4 d F,U 8 12 17.78 76.33 Diamond et al. 1993

Spring peeper,

Pseudacris crucifer Ammonium

chloride 4 d F,U 8 20 11.42 49.04 61.18 Diamond et al. 1993

Pacific tree frog (embryo),

Pseudacris regilla Ammonium

nitrate 4 d R,M 6.7 22 41.19 33.36 Schuytema and Nebeker

1999a

## 123

Appendix A. Acute Toxicity of Ammonia to Aquatic Animals

Species Chemical Name Duration Methods a pH Temp.

( ̊C) Total

Ammonia

(mg

TA N/L) Total Ammonia b

(mg TA N/L)

a djusted to pH 7

(all organisms )

and 20 ° C

( invertebrates ) SMAV

(mg

TA N/L) GMAV

(mg

TA N/L) Reference

Pacific tree frog (embryo),

Pseudacris regilla Ammonium

chloride 4 d R,M 6.7 22 60.44 48.95 Schuytema and Nebeker

1999a

Pacific tree frog (embryo),

Pseudacris regilla Ammonium

sulfate 4 d R,M 6.7 22 103.1 83.53 Schuytema and Nebeker

1999a

Pacific tree frog

(90 mg, Gosner Stage 26

- 27),

Pseudacris regilla Nitric acid

ammonium salt 4 d R,M 7.3 22 136.6 188.1 Schuytema and Nebeker

1999b

Pacific tree frog

(60 mg, Gosner Stage 26

- 27),

Pseudacris regilla Ammonium

sulfate 4 d R,M 7.3 22 116.4 160.2 83.71 71.56 Schuytema and Nebeker

1999b

Mucket (glochidia),

Actinonaias ligamentina Ammonium

chloride 1 d S,M 8.6 20 6.141 c 83.61 Wang et al. 2007b

Mucket (glochidia),

Actinonaias ligamentina Ammonium

chloride 1 d S,M 8.4 20 8.099 c 75.29 Wang et al. 2007b

Mucket (glochidia),

Actinonaias ligamentina Ammonium

chloride 1 d S,M 8.3 20 5.073 c 38.84 Wang et al. 2007b

Mucket (glochidia),

Actinonaias ligamentina Ammonium

chloride 1 d S,M 8.3 20 8.900 c 68.13 63.89 Wang et al. 2007b

Pheasantshell (juvenile),

Actinonaias pectorosa Ammonium

chloride 4 d S,M 7.9 25 14.06 75.80 Keller 2000

Pheasantshell (juvenile),

Actinonaias pectorosa Ammonium

chloride 4 d S,M 7.95 25 14.08 83.30 79.46 71.25 Keller 2000

Giant floater mussel (adult),

Pyganodon grandis Ammonium

chloride 4 d S,M 7.71 25 18.84 72.49 Scheller 1997

Giant floater mussel (adult),

Pyganodon grandis Ammonium

chloride 4 d S,M 7.5 25 25.13 69.02 70.73 70.73 Scheller 1997

Shortnose sucker (0.53

- 2.00 g),

Chasmistes brevirostris Ammonium

chloride 4 d F,M 8 20 11.42 49.04 Saiki et al. 1999

Shortnose sucker,

Chasmistes brevirostris Ammonium

chloride 4 d F,M 8 20 22.85 98.09 69.36 69.36 Saiki et al. 1999

## 124

Appendix A. Acute Toxicity of Ammonia to Aquatic Animals

Species Chemical Name Duration Methods a pH Temp.

( ̊C) Total

Ammonia

(mg

TA N/L) Total Ammonia b

(mg TA N/L)

a djusted to pH 7

(all organisms )

and 20 ° C

( invertebrates ) SMAV

(mg

TA N/L) GMAV

(mg

TA N/L) Reference

Pagoda hornsnail (adult),

Pleurocera uncialis Ammonium

chloride 4 d R,M 8.1 22 11.18 68.54 68.54 68.54 Goudreau et al. 1993

Golden shiner,

Notemigonus crysoleucas Ammonium

chloride 4 d S,M 7.5 19.6 89.61 162.6 EA Engineering 1985

Golden shiner,

Notemigonus crysoleucas Ammonium

chloride 4 d S,M 7.55 19.5 73.85 144.6 EA Engineering 1985

Golden shiner (8.7 g),

Notemigonus crysoleucas Ammonium

chloride 4 d F,M 7.5 24.5 34.73 63.02 63.02 63.02 Swigert and Spacie 1983

Pebblesnail (1.8 mm),

Fluminicola sp. Ammonium

chloride 4 d F,M 8.25 20.2 >8.801 >62.15 >62.15 >62.15 Besser 2011

Lost River sucker (0.49

- 0.80 g),

Deltistes luxatus Ammonium

chloride 4 d F,M 8 20 16.81 72.18 Saiki et al. 1999

Lost River sucker (larvae),

Deltistes luxatus Ammonium

chloride 4 d F,M 8 20 10.35 44.42 56.62 56.62 Saiki et al. 1999

Mountain whitefish

(177 g, 27.0 cm),

Prosopium williamsoni Ammonium

chloride 4 d F,U 7.68 12.1 11.3 27.31 Thurston and Meyn 1984

Mountain whitefish

(56.9 g, 19.1 cm),

Prosopium williamsoni Ammonium

chloride 4 d F,U 7.84 12.4 25.47 81.35 Thurston and Meyn 1984

Mountain whitefish

(63.0 g, 20.4 cm),

Prosopium williamsoni Ammonium

chloride 4 d F,U 7.8 12.3 21.2 63.04 51.93 51.93 Thurston and Meyn 1984

Atlantic pigtoe (glochidia),

Fusconaia masoni Ammonium

chloride 6 h S,M 7.6 24.9 15.9 47.40 47.40 47.40 Black 2001

Pondshell mussel (juvenile),

Utterbackia imbecillis Ammonium

chloride 4 d S,M 7.9 24 8.235 40.87 Keller 2000

Pondshell mussel (juvenile),

Utterbackia imbecillis Ammonium

chloride 4 d S,M 8.35 25 3.269 41.75 Keller 2000

## 125

Appendix A. Acute Toxicity of Ammonia to Aquatic Animals

Species Chemical Name Duration Methods a pH Temp.

( ̊C) Total

Ammonia

(mg

TA N/L) Total Ammonia b

(mg TA N/L)

a djusted to pH 7

(all organisms )

and 20 ° C

( invertebrates ) SMAV

(mg

TA N/L) GMAV

(mg

TA N/L) Reference

Pondshell mussel (juvenile),

Utterbackia imbecillis Ammonium

chloride 4 d S,M 7.9 25 9.355 50.45 Keller 2000

Pondshell mussel

(8 d old juvenile),

Utterbackia imbecillis Ammonium

chloride 4 d R,M 7.8 24 14.29 59.19 Wade et al. 1992

Pondshell mussel

(<2 d old juvenile),

Utterbackia imbecillis Ammonium

chloride 4 d R,M 8.16 25 5.254 46.38 Black 2001

Pondshell mussel

(<2 d old juvenile),

Utterbackia imbecillis Ammonium

chloride 4 d R,M 8.17 25 5.781 52.03 Black 2001

Pondshell mussel

(<2 d old juvenile),

Utterbackia imbecillis Ammonium

chloride 4 d R,M 8.29 25 8.845 100.5 Black 2001

Pondshell mussel

(<2 d old juvenile),

Utterbackia imbecillis Ammonium

chloride 4 d R,M 8 25.1 2.734 17.91 Black 2001

Pondshell mussel (glochidia),

Utterbackia imbecillis Ammonium

chloride 1 d S,M 8.02 25 7.395 49.90 46.93 46.93 Black 2001

Pink mucket

(2 mo old juvenile),

Lampsilis abrupta Ammonium

chloride 4 d R,M 8.3 20 1.921 d 14.71 Wang et al. 2007b

Pink mucket

(2 mo old juvenile),

Lampsilis abrupta Ammonium

chloride 4 d F,M 8.4 20 2.8 26.03 26.03 Wang et al. 2007a

Plain pocketbook

(3

- 5 d old juvenile),

Lampsilis cardium Ammonium

chloride 4 d S,M 8.2 20.5 23.50 e 154.4 Newton et al. 2003

Plain pocketbook

(3

- 5 d old juvenile),

Lampsilis cardium Ammonium

chloride 4 d S,M 8.2 21.2 23.70 e 165.0 Newton et al. 2003

Plain pocketbook

(1

- 2 d old juvenile),

Lampsilis cardium Ammonium

chloride 4 d F,M 7.6 21.2 23.1 54.07 Newton and Bartsch 2007

## 126

Appendix A. Acute Toxicity of Ammonia to Aquatic Animals

Species Chemical Name Duration Methods a pH Temp.

( ̊C) Total

Ammonia

(mg

TA N/L) Total Ammonia b

(mg TA N/L)

a djusted to pH 7

(all organisms )

and 20 ° C

( invertebrates ) SMAV

(mg

TA N/L) GMAV

(mg

TA N/L) Reference

Plain pocketbook

(1

- 2 d old juvenile),

Lampsilis cardium Ammonium

chloride 4 d F,M 7.1 21.2 38.9 47.19 50.51 Newton and Bartsch 2007

Wavy

- rayed lampmussel

(2

- 5 d old juvenile),

Lampsilis fasciola Ammonium

chloride 4 d R,M 7.83 12.6 14.9 25.31 Mummert et al. 2003

Wavy

- rayed lampmussel

(<5 d old juvenile),

Lampsilis fasciola Ammonium

chloride 4 d R,M 8.5 20 6.179 d 69.63 Wang et al. 2007b

Wavy

- rayed lampmussel

(glochidia),

Lampsilis fasciola Ammonium

chloride 1 d S,M 8.3 20 7.743 c 59.28 Wang et al. 2007b

Wavy

- rayed lampmussel

(glochidia),

Lampsilis fasciola Ammonium

chloride 1 d S,M 8.4 20 5.518 c 51.30 48.11 Wang et al. 2007b

Higgin's eye

(1

- 2 d old juvenile),

Lampsilis higginsii Ammonium

chloride 4 d F,M 7.6 21.2 19.5 45.64 Newton and Bartsch 2007

Higgin's eye

(1

- 2 d old juvenile),

Lampsilis higginsii Ammonium

chloride 4 d F,M 7.1 21.2 31.7 38.46 41.90 Newton and Bartsch 2007

Neosho mucket

(<5 d old juvenile),

Lampsilis rafinesqueana Ammonium

chloride 4 d R,M 8.3 20 9.185 d 70.31 Wang et al. 2007b

Neosho mucket

(<5 d old juvenile),

Lampsilis rafinesqueana Ammonium

chloride 4 d R,M 8.4 20 9.269 d 86.17 Wang et al. 2007b

Neosho mucket (glochidia),

Lampsilis rafinesqueana Ammonium

chloride 1 d S,M 8.3 20 7.387 c 56.55 69.97 Wang et al. 2007b

Fatmucket (juvenile),

Lampsilis siliquoidea Ammonium

chloride 4 d S,M 8.3 24 1.275 13.60 Myers

- Kinzie 1998

## 127

Appendix A. Acute Toxicity of Ammonia to Aquatic Animals

Species Chemical Name Duration Methods a pH Temp.

( ̊C) Total

Ammonia

(mg

TA N/L) Total Ammonia b

(mg TA N/L)

a djusted to pH 7

(all organisms )

and 20 ° C

( invertebrates ) SMAV

(mg

TA N/L) GMAV

(mg

TA N/L) Reference

Fatmucket (3 mo old juvenile),

Lampsilis siliquoidea Ammonium

chloride,

ammonium

hydroxide 4 d F,M 8.35 20 8.80 74.25 Miao et al. 2010

Fatmucket (2 mo old juvenile),

Lampsilis siliquoidea Ammonium

chloride 4 d R,M 8.1 20 4.092 d 21.26 Wang et al. 2007b

Fatmucket (2 mo old juvenile),

Lampsilis siliquoidea Ammonium

chloride 4 d F,M 8.2 20 4.6 28.99 Wang et al. 2007a

Fatmucket (7 d old juvenile),

Lampsilis siliquoidea Ammonium

chloride,

ammonium

hydroxide 4 d F,M 7.6 20.5 11 24.30 Wang et al. 2008

Fatmucket (7 d old juvenile),

Lampsilis siliquoidea Ammonium

chloride,

ammonium

hydroxide 4 d F,M 8.1 20.6 5.2 28.39 Wang et al. 2008

Fatmucket (7 d old juvenile),

Lampsilis siliquoidea Ammonium

chloride,

ammonium

hydroxide 4 d F,M 8.5 20.6 3.4 40.27 Wang et al. 2008

Fatmucket (7 d old juvenile),

Lampsilis siliquoidea Ammonium

chloride,

ammonium

hydroxide 4 d F,M 9 20.6 0.96 27.51 Wang et al. 2008

Fatmucket (7 d old juvenile),

Lampsilis siliquoidea Ammonium

chloride,

ammonium

hydroxide 4 d F,M 6.6 19.6 88 65.59 Wang et al. 2008

Fatmucket (7 d old juvenile),

Lampsilis siliquoidea Ammonium

chloride,

ammonium

hydroxide 4 d F,M 8.1 19.4 11 54.37 Wang et al. 2008

Fatmucket (<5 d old juvenile),

Lampsilis siliquoidea Ammonium

chloride 4 d R,M 8.5 20 8.350 d 94.09 Wang et al. 2007b

Fatmucket (glochidia),

Lampsilis siliquoidea Ammonium

chloride 1 d S,M 8.4 20 9.790 c 91.01 Wang et al. 2007b

## 128

Appendix A. Acute Toxicity of Ammonia to Aquatic Animals

Species Chemical Name Duration Methods a pH Temp.

( ̊C) Total

Ammonia

(mg

TA N/L) Total Ammonia b

(mg TA N/L)

a djusted to pH 7

(all organisms )

and 20 ° C

( invertebrates ) SMAV

(mg

TA N/L) GMAV

(mg

TA N/L) Reference

Fatmucket (glochidia),

Lampsilis siliquoidea Ammonium

chloride 1 d S,M 8.2 20 13.35 c 84.14 Wang et al. 2007b

Fatmucket (glochidia),

Lampsilis siliquoidea Ammonium

chloride 1 d S,M 8.4 20 11.57 c 107.6 Wang et al. 2007b

Fatmucket (glochidia),

Lampsilis siliquoidea Ammonium

chloride 1 d S,M 8.5 20 >14.24 c 160.5 Wang et al. 2007b

Fatmucket (glochidia),

Lampsilis siliquoidea Ammonium

chloride 1 d S,M 8.3 20 6.497 c 49.74 Wang et al. 2007b

Fatmucket (glochidia),

Lampsilis siliquoidea Ammonium

chloride 1 d S,M 8.3 20 8.772 c 66.77 55.42 46

. 63 Wang et al. 2007b

Rainbow mussel

(2 mo old juvenile),

Villosa iris Ammonium

chloride 4 d R,M 8.4 20 2.505 d 23.29 Wang et al. 2007b

Rainbow mussel

(2 mo old juvenile),

Villosa iris Ammonium

chloride 4 d R,M 8.3 20 8.935 d 68.40 Wang et al. 2007b

Rainbow mussel

(5 d old juvenile),

Villosa iris Ammonium

chloride 4 d S,M 8.18 25 7.81 71.66 Scheller 1997

Rainbow mussel

(<5 d old juvenile),

Villosa iris Ammonium

chloride 4 d R,M 8.1 20 5.261 d 27.33 Wang et al. 2007b

Rainbow mussel

(2

- 5 d old juvenile),

Villosa iris Ammonium

chloride 4 d R,M 7.29 12.6 20.6 15.17 Mummert et al. 2003

Rainbow mussel

(<3 d old juvenile),

Villosa iris Ammonium

chloride 4 d S,M 8.18 25 7.07 64.87 Scheller 1997

Rainbow mussel

( < 24 h old glochidia),

Villosa iris Ammonium

chloride 1 d S,M 7.94 20.0 3.290 12.62 Scheller 1997

Rainbow mussel (glochidia),

Villosa iris Ammonium

chloride 1 d S,M 8.4 20 10.68 c 99.28 Wang et al. 2007b

## 129

Appendix A. Acute Toxicity of Ammonia to Aquatic Animals

Species Chemical Name Duration Methods a pH Temp.

( ̊C) Total

Ammonia

(mg

TA N/L) Total Ammonia b

(mg TA N/L)

a djusted to pH 7

(all organisms )

and 20 ° C

( invertebrates ) SMAV

(mg

TA N/L) GMAV

(mg

TA N/L) Reference

Rainbow mussel

(<1 h old glochidia),

Villosa iris Ammonium

chloride 1 d R,M 8.1 22 3.570 21.89 Goudreau et al. 1993

Rainbow mussel

(<1 h old glochidia),

Villosa iris Ammonium

chloride 1 d R,M 8.1 22 4.278 26.23 34.23 34.23 Goudreau et al. 1993

Oyster mussel

(<5 d old juvenile),

Epioblasma capsaeformis Ammonium

chloride 4 d R,M 8.5 20 4.760 d 53.63 Wang et al. 2007b

Oyster mussel (glochidia),

Epioblasma capsaeformis Ammonium

chloride 6 h R,M 8.5 20 5.0 c 17.81 Wang et al. 2007b

Oyster mussel (glochidia),

Epioblasma capsaeformis Ammonium

chloride 6 h R,M 8.5 20 3.4 c 31.61 31.14 31.14 Wang et al. 2007b

Green floater (<2 d old

juvenile),

Lasmigona subviridis Ammonium

chloride 4 d R,M 7.73 24 6.613 24.24 Black 2001

Green floater (<2 d old

juvenile),

Lasmigona subviridis Ammonium

chloride 4 d R,M 7.73 24 6.613 24.24 Black 2001

Green floater (<2 d old

juvenile),

Lasmigona subviridis Ammonium

chloride 4 d R,M 7.92 24.8 3.969 21.84 23.41 23.41 Black 2001

Ellipse (glochidia),

Venustaconcha ellipsiformis Ammonium

chloride 1 d S,M 8.1 20 4.550 c 23.12 23.12 23.12 Wang et al. 2007b

a S = static, R = renewal, F = flow

- through, and NR= not reported (uncertain) exposure types; M = measured and U = unmeasured tests.

b Acute values are normalized to pH 7 (all organisms) and temperature 20 ° C (invertebrates) as per the equations provided in this document (see also 1999 AWQC document for the basis of

the pH

- and temperature

- dependence of ammonia toxicity and Appendix D for an example calculation).

c The EC 50 s reported in this study were based on nominal concentrations. Percent nominal concentrations of measured ammonia concent rations on exposure days 0 and 2 declined from 104

to 44 . EC 50 s based on measured concentrations were estimated from the reported EC 50 s based on nominal concen trations by multiplying by 0.890 for the 24 hr test; this factor is the average

of the percent nominal concentrations of measured concentrations from ammonia measuremen ts made on exposure day 0 (i.e., 104) and estimated for day 1 (i.e, 74) of the study.

## 130

d The EC 50 s re ported in this study were based on nominal concentrations. Percent nominal concentrations of measured ammonia concentrations on exposure days 0 and 4 declined from 104

to 63. EC 50 s based on measured concentrations were estimated from the reported EC 50 s b ased on nominal concentrations by multiplying by 0.835 or the average of the percent nominal

concentrations of measured concentrations from ammonia measurements made on exposure days 0 and 4 in the study.

e EC 50 values based on sediment porewater concentrations. Note : these EC 50 s were not used to calculate the SMAV for the species.

f This small subset of LC 50 s for adult rainbow trout from Thurston and Russo (1983) was used as the basis for the FAV calculated in the 1999 AWQC document. The FAV in the 1999

AWQC docume nt of 11.23 mg TAN/L at pH 8 was lowered to the geometric mean of these five LC 50 values at the time in order to protect large rainbow trout

, which were shown to be

measurably more sensitive than other life stages. The FAV prior to adjusting it to protect the commercially and recreationally important adult rainbow trout was calculated to be 14.32 mg

TA N/L (CMC = 7.2 mg TA N/L ) at pH 8. Th is FAV based on protection of adult rainbow trout at pH 7 is 48.2 1 mg TA N/L (see Table 7 in this document)

. Because several equivalent LC 50 s

representing different ages and life

- stages have been added to the current (updated) acute criteria dataset, it no longer seems appropriate to lower the SMAV for rainbow trout base d on only

these five LC 50 s considering the several other additional acute values which now exist.

Note : Each SMAV was calculated from the associated bold

- face number(s) in the preceding column.

## 131

Appendix B. Chronic Toxicity of Ammonia to Aquatic Animals.

Appendix B. Chronic Toxicity of Ammonia to Aquatic Animals

Species Test and Effect pH Temp.

( ̊C) Total

Ammonia

(mg

TA N/L) Chronic value a

Adjusted to pH 7

(all organisms )

and 20 ° C

( invertebrates )

(mg TA N/L) SMCV

(mg

TA N/L) GMCV

(mg

TA N/L) Reference

Stonefly,

Pteronarcella badia 30

- d Juv Survival 8.04 12.1 133.8 207.0 Thurston et al. 1984b

Stonefly,

Pteronarcella badia 24

- d Juv Survival 7.81 13.2 21.66 26.27 73.74 73.74 Thurston et al. 1984b

Water flea,

Ceriodaphnia acanthina 7

- d LC Reproduction 7.15 24.5 44.90 64.10 64.10 Mount 1982

Water flea,

Ceriodaphnia dubia 7

- d LC Reproduction 7.80 25.0 15.20 38.96 Nimmo et al. 1989

Water flea,

Ceriodaphnia dubia 7

- d LC Reproduction 8.57 26.0 5.800 52.15 45.08 53.75 Willingham 1987

Water flea,

Daphnia magna 21

- d LC Reproduction 8.45 19.8 7.370 36.27 Gersi ch et al. 1985

Water flea,

Daphnia magna 21

- d LC Reproduction 7.92 20.1 21.70 47.40 41.46 41.46 Reinbold and Pescitelli

1982a

Amphipod,

Hyalella azteca 28

- d PLC Biomass 8.04 25.0 8.207 29.17 29.17 29.17 Borgmann 1994

Channel catfish,

Ictalurus punctatus 30

- d ELS Weight 7.80 25.8 12.20 22.66 Reinbold and Pescitelli

1982a

Channel catfish,

Ictalurus punctatus 30

- d Juv Survival 8.35 27.9 5.020 21.15 Colt and Tchobanoglous

1978

Channel catfish,

Ictalurus punctatus 30

- d ELS Biomass 7.76 26.9 11.50 20.35 21.36 21.36 Swigert and Spacie 1983

Northern pike (fertilized),

Esox lucius 52

- d ELS Biomass 7.62 8.70 13.44 20.38 20.38 20.38 Harrahy et al. 2004

Common carp (fertilized),

Cyprinus carpio 28

- d ELS Weight 7.85 23.0 8.360 16.53 16.53 16.53 Mallet and Sims 1994

## 132

Appendix B. Chronic Toxicity of Ammonia to Aquatic Animals

Species Test and Effect pH Temp.

( ̊C) Total

Ammonia

(mg

TA N/L) Chronic value a

Adjusted to pH 7

(all organisms )

and 20 ° C

( invertebrates )

(mg TA N/L) SMCV

(mg

TA N/L) GMCV

(mg

TA N/L) Reference

Lahontan cutthroat trout (fertilized),

Oncorhynchus clarkii henshawi 103

- d ELS Survival 7.57 13.7 17.89 25.83 25.83 Koch et al. 1980

Rainbow trout (fertilized),

Oncorhynchus mykiss 42

- d ELS Survival 7.50 10.0 <33.6 <45.5 Burkhalter and Kaya 1977

Rainbow trout,

Oncorhynchus mykiss 72

- d ELS Survival 7.40 14.5 2.600 3.246 Calamari et al. 1977, 1981

Rainbow trout (fertilized),

Oncorhynchus mykiss 73

- d ELS Survival 7.52 14.9 <2.55 <3.515 Solbe and Shurben 1989

Rainbow trout,

Oncorhynchus mykiss 5

- year LC 7.70 7.5

-

10.5 >6.71 >11.08 Thurston et al. 1984 a

Rainbow trout,

Oncorhynchus mykiss 90

- d ELS Survival 7.75 11.4 8.919 15.60 6.663 Brinkman et al. 2009

Sockeye salmon,

Oncorhynchus nerka 62

- d Embryos

Hatchability 8.42 10.0 <2.13 <10.09 10.09 12.02 Rankin 1979

White sucker (3 d old embryo),

Catostomus commersonii 30

- d ELS Biomass 8.32 18.6 2.900 >11.62 11.62 11.62 Reinbold and Pescitelli

1982a

Smallmouth bass,

Micropterus dolomieu 32

- d ELS Biomass 6.60 22.3 9.610 8.650 Broderius et al. 1985

Smallmouth bass,

Micropterus dolomieu 32

- d ELS Biomass 7.25 22.3 8.620 9.726 Broderius et al. 1985

Smallmouth bass,

Micropterus dolomieu 32

- d ELS Biomass 7.83 22.3 8.180 15.77 Broderius et al. 1985

Smallmouth bass,

Micropterus dolomieu 32

- d ELS Biomass 8.68 22.3 1.540 11.31 11.07 11.07 Broderius et al. 1985

Fathead minnow (embryo

- larvae),

Pimephales promelas 28

- d ELS Survival 8.00 24.8 5.120 12.43 Mayes et al. 1986

Fathead minnow (embryo

- larvae),

Pimephales promelas 32

- d ELS Biomass 7.95 25.5 7.457 16.87 Adelman et al. 2009

Fathead minnow,

Pimephales promelas 30

- d ELS Biomass 7.82 25.1 3.730 7.101 Swigert and Spacie 1983

## 133

Appendix B. Chronic Toxicity of Ammonia to Aquatic Animals

Species Test and Effect pH Temp.

( ̊C) Total

Ammonia

(mg

TA N/L) Chronic value a

Adjusted to pH 7

(all organisms )

and 20 ° C

( invertebrates )

(mg TA N/L) SMCV

(mg

TA N/L) GMCV

(mg

TA N/L) Reference

Fathead minnow,

Pimephales promelas LC Hatchability 8.00 24.2 1.970 4.784 9.187 9.187 Thurston et al. 1986

Pebblesnail (1.81 mm

, juvenile),

Fluminicola sp. 28

- d Juv Change in

Length 8.22 20.1 2.281 7.828 7.828 7.828 Besser 2011

Long fingernailclam,

Musculium transversum 42

- d Juv Survival 8.15 23.5 5.820 22.21 Anderson et al. 1978

Long fingernailclam,

Musculium transversum 42

- d Juv Survival 7.80 21.8 1.230 2.565 7.547 7.547 Sparks and Sandusky 1981

Green sunfish,

Lepomis cyanellus 30

- d ELS Biomass 7.90 22.0 5.610 11.85 McCormick et al. 1984

Green sunfish,

Lepomis cyanellus 30

- d ELS Survival 8.16 25.4 5.840 18.06 14.63 Reinbold and Pescitelli

1982a

Bluegill,

Lepomis macrochirus 30

- d ELS Biomass 7.76 22.5 1.850 3.273 3.273 6.920 Smith et al. 1984

Rainbow mussel (2 mo old juvenile),

Villosa iris 28

- d Juv Survival 8.20 20.0 1.063 3.501 3.501 3.501 Wang et al. 2007a

Fatmucket (2 mo old juvenile),

Lampsilis siliquoidea 28

- d Juv Survival 8.2 5 20.0 0. 8988 3.211 3.211 Wang et al. 200 11

Wavy

- rayed lamp mussel

(2 mo old juvenile),

Lampsilis fasciola 28

- d Juv Survival 8.20 20.0 0. 4272 1.408 1. 408 2.126 Wang et al. 2007a

a The chronic value is an EC 20 value calculated using EPA’s TRAP (Versions 1.0 or 1.2 1 a ). Note: all chronic values were normalized to pH 7 (all organisms) and 2 0 ° C

(invertebrates) as per the equations provided in this document (see also 1999 AWQC document for the basis of the pH

- and temperature

- dependence of ammonia toxicity and

Appendix E for an example calculation).

Note: Each SMC V was calculated from the associated bold

- face number(s) in the preceding column.

## 134

Appendix C. Other Chronic Ammonia Toxicity Data.

Appendix C. Other Chronic Ammonia Toxicity Data

Species Test and Effect Method a pH Temp.

( ̊C) Total

Ammonia

(mg

TAN/L) Chronic value

Adjusted to pH 7

(all organisms ) and

20°C ( invertebrates )

(mg TAN/L) Reference

FRESHWATER INVERTEBRATES

Pulmonate pondsnail

(<1 wk post hatch),

Lymnaea stagnalis 28

- d NOEC

-

Growth F,M 8.25 20.1 >8.00 >28.76 Besser et al. 2009

Pulmonate pondsnail

(<1 wk post

- hatch),

Lymnaea stagnalis 28

- d NOEC

-

Survival F,M 8.25 20.1 >8.00 >28.76 Besser et al. 2009

Idaho springsnail

(7

- 9 and 11

- 13 wk post hatch juvenile),

Pyrgulopsis idahoensis 28

- d NOEC

-

Growth F,M 8.25 20.1 >8.00 >28.76 Besser et al. 2009

Idaho springsnail

(7

- 9 and 11

- 13 wk post hatch juvenile),

Pyrgulopsis idahoensis 28

- d EC 20 - Survival F,M 8.25 20.1 0.480 1.726 Besser et al. 2009

Idaho springsnail

(mixed

- aged, adults),

Pyrgulopsis idahoensis 28

- d EC 20 - Survival F,M 8.26 20.8 3.24 12.39 b Besser et al. 2009

Pebblesnail (mixed

- aged, field collected),

Fluminicola sp. 28

- d EC 20 - Survival F,M 8.26 20.8 1.02 3.900 c Besser et al. 2009

Pebblesnail (small, field collected),

Fluminicola sp. 28

- d MATC

-

Survival F,M 8.19 20.1 2.75 8.977 d Besser 2011

Ozark springsnail

(mixed age, field collected),

Fontigens aldrichi 28

- d EC 20 - Survival F,M 8.26 20.8 0.61 2.332 b Besser et al. 2009

Bliss Rapids snail

(mixed age, field collected),

Taylorconcha serpenticola 28

- d EC 20 - Survival F,M 8.26 20.8 3.42 13.08 b Besser et al. 2009

## 135

Appendix C. Other Chronic Ammonia Toxicity Data

Species Test and Effect Method a pH Temp.

( ̊C) Total

Ammonia

(mg

TAN/L) Chronic value

Adjusted to pH 7

(all organisms ) and

20°C ( invertebrates )

(mg TAN/L) Reference

Silty hornsnail

(mixed age, mature and field collected),

Pleurocera canaliculata 28

- d EC 20 - Survival;

(Alt Effect Conc.) F,M 8.15 24.7 0.45

( ≤ 1.86) 1.845

(≤7.667) b

, e GLEC 2011

Wavy

- rayed lamp mussel

(2 mo old juvenile),

Lampsilis fasciola 28

- d IC 25 - Growth F,M 8.20 20.0 0.5700 1.878 Wang et al. 2007a

Fatmucket (2 mo old juvenile),

Lampsilis siliquoidea 28

- d IC 25 - Growth F,M 8.20 20.0 0.4400 1.450 Wang et al. 2007a

Rainbow mussel (2 mo old juvenile),

Villosa iris 28

- d IC 25 - Growth F,M 8.20 20.0 0.7300 2.406 Wang et al. 2007a

Water flea, (<24 hr),

Ceriodaphnia dubia 7

- d;

3 broods in control

IC 25 Reproduction R,U 7.90 25.0 1.300 3.790 Dwyer et al. 2005

FRESHWATER VERTEBRATES

Cutthroat trout (3.3 g),

Oncorhynchus clarkii 29

- d LC 50 F,M 7.80 12.4 21.60 40.11 Thurston et al. 1978

Cutthroat trout (3.4 g),

Oncorhynchus clarkii 29

- d LC 50 F,M 7.78 12.2 21.40 38.78 Thurston et al. 1978

Cutthroat trout (1.0 g),

Oncorhynchus clarkii 36

- d LC 50 F,M 7.81 13.1 30.80 57.91 Thurston et al. 1978

Cutthroat trout (1.0 g),

Oncorhynchus clarkii 36

- d LC 50 F,M 7.80 12.8 32.20 59.79 Thurston et al. 1978

Atlantic salmon,

Salmo salar 105

- d Juv NOEC

-

Survival F,M 6.84 12.1 >32.29 >30.64 Kolarevic et al. 2012

Lake trout, siscowet,

Salvelinus namaycush 60

- d LOEC

Weight gain F,M 8.02 11.6 6.440 16.10 Beamish and Tandler 1990

Brook trout (juvenile),

Salvelinus fontinalis 4

- d Juv LOEC

- Swimming Perf F,M 9.10 15.0 0.7765 10.86 Tudorache et al. 201 0

Bonytail chub

(2 and 7 d post hatch),

Gila elegans 7

- d IC 25 - Growth R,U 7.90 25.0 11.00 23.24 Dwyer et al. 2005

## 136

Appendix C. Other Chronic Ammonia Toxicity Data

Species Test and Effect Method a pH Temp.

( ̊C) Total

Ammonia

(mg

TAN/L) Chronic value

Adjusted to pH 7

(all organisms ) and

20°C ( invertebrates )

(mg TAN/L) Reference

Spotfin chub (<24 hr),

Erimonax monachus 7

- d IC 25 - Growth R,U 7.90 25.0 15.80 33.37 Dwyer et al. 2005

Cape Fear shiner (<24 hr),

Notropis mekistocholas 7

- d IC 25 - Growth R,U 7.90 25.0 8.800 18.59 Dwyer et al. 2005

Topeka shiner (adult),

Notropis topeka 30

- d EC 20 - Survival F,M 7.94 23.9 10.85 24.21 Adelman et al. 2009

Topeka shiner (juvenile, 11 mo),

Notropis topeka 30

- d EC 20 - SGR F,M 8.07 12.4 6.483 17.45 Adelman et al. 2009

Gila topminnow (<24, 48 and 72 hr),

Poeciliopsis occidentalis 7

- d IC 25 - Growth R,U 7.90 25.0 24.10 50.91 Dwyer et al. 2005

Fathead minnow (<24 hr),

Pimephales promelas 7

- d IC 25 - Growth R,U 7.90 25.0 7.200 15.21 Dwyer et al. 2005

Fathead minnow (4 d post hatch),

Pimephales promelas 28

- d LOEC

Survival R,M 8.25 19.9 9.160 32.71 Fairchild et al. 2005

Colorado pikeminnow

(5 and 6

- d post hatch),

Ptychocheilus lucius 7

- d IC 25 - Growth R,U 7.90 25.0 8.900 18.80 Dwyer et al. 2005

Colorado pikeminnow (juvenile, 8 d),

Ptychocheilus lucius 28

- d LOEC

- Growth R,M 8.23 19.9 8.600 29.75 Fairchild et al. 2005

Razorback sucker (7 d post hatch),

Xyrauchen texanus 7

- d IC 25 - Growth R,U 7.90 25.0 13.40 28.30 Dwyer et al. 2005

Razorback sucker (9 d),

Xyrauchen texanus 28

- d LOEC

Survival R,M 8.24 19.9 13.25 46.58 Fairchild et al. 2005

Lost River sucker (late

- stage larva),

Deltistes luxatus 30

- d LOEC

Survival F,M 9.43 22.3 1.230 25.31 Meyer and Hansen 2002

Green frog (Stage 24

- 26),

Rana clamitans 103

- d NOEC

Growth R,M 8.70 24.0 >2.20 >16.74 Jofre and Karasov 1999

a R = renewal and F = flow

- through exposure types; M = measured and U = unmeasured tests.

b Not used in the calculation of the SMCV because of the uncertainty of the chronic value, but included here as weight of evide nce supporting the sensitivity of non

- pulmonate

snail species in general as determined by 28

- day toxicity tests (see Additional 28

- day Toxicity test Data for Freshwater Snails in Appendix I for more detail ).

c Not used in the calculation of the SMCV because of the uncertainty of the chronic value, but included here as weight of evide nce supporting the sensitivity of non

- pulmonate

snai l species in general as determined by 28

- day toxicity tests (see Chronic Toxicity T est Data : 28

- day Tests with Juvenile and Adult Pebblesnails in Appendix H for more detail ).

## 137 d Not used in the calculation of the SMCV because of low control survival (75 per cent) for this size class.

e Value represents a 28

- day ammonia survival effects concentration used in place of the EC 20 due to the high degree of temporal variability in measured total ammonia

concentrations in the test, as well as the unequal response amo ng test replicates near this concentration (see Additional 28

- day Toxicity test Data for Freshwater Snails in Appendix

I for more detail ).

## 138

Appendix D. Conversion of Acute Results of Toxicity Tests

.

All of the ammonia acute values (LC 50 

## s and EC 50 

## s) in Appendix A of this document were

converted to TAN acute values using the reported temperatures and pHs, and using the pKa

relationship from Emerson et al. (1975). Conversions were dependent on the form of ammonia

the acute values were expre ssed, e.g., unionized ammonia (UIA), unionized ammonia expressed

as nitrogen (UIA

- N), total ammonia (TA) and total ammonia nitrogen (TAN). After acute values

were converted to TAN they were then normalized to pH 7 using the pH relationship developed

in th e 1999 AWQC document. Following the adjustment to pH 7 , the TAN acute values were

further normalized to a temperature of 2 0 ° 

## C for invertebrates only, following recommendations

in the 1999 AWQC document. It is worth noting here that while the relationship between pH

and ammonia toxicity was first addressed in the 1985 criteria document, it was not fully

developed until the 1999 AWQC update document. Detailed information regarding the

development and parameterization of the pH

- ammonia toxicity equations (a cute and chronic) can

be found in the 1999 AWQC document (pH

- Dependence of Ammonia Toxicity

– U.S. EPA

1999). In contrast to the pH

- toxicity relationship

, which applies to both vertebrates and

invertebrates, the temperature

- ammonia toxicity relationship o nly applies to invertebrates. Based

on the results of the 1999 reanalysis of this relationship, it was determined that ammonia toxicity

for invertebrates decreases with decreasing temperature to a temperature of approximately 7°C,

below which the relations hip ends (U.S. EPA 1999).

The conversion procedure for acute toxicity values is illustrated here using the data for

the flatworm, Dendrocoelum lacteum , which is the first species listed in Table 1 in the

1984/1985 criteria document and was the species chosen to illustrate the conversion procedure in

Appendix 3 of the 1999 AWQC document:

Acute value (AV) = 1.40 mg unionized ammonia (UIA) or NH 3 

## /L

Test pH = 8.20

Test Temperature = 18.0 ° C

## 139

Step 1.

Equation 3 in the 1999 criterion document , and the Emerson et al. (1975) equation from

page 7 of this document, is used to calculate the pK a at 18 ° C:

pKa = 9.464905

Step 2

.

The AV in terms of total ammonia (TA) is calculated as:

[NH 3 

## ]/[NH +

4 

## ] = 10 (pH

- pK) 

## = 0.0543369

Step 3

.

The Wood (1993) equation from page 7 ( Equation 2 in the 1999 AWQC document ) is

rearranged to obtain the acute value for TA :

TA = [NH 3 

## ] + [NH +

4 

## ] = [NH 3 

## ] + [NH 3 

## ]/ (10 (pH

- p Ka ) 

## )

TA = [NH 3 

## ] + [NH +

4 

## ] = [NH 3 

## ] + [NH 3 

## ]/0.0543369

= 27.1652 mg TA /L

Step 4.

The AV for TA is converted to the AV for TAN (AV

t 

## ) as follows:

AV

t 

## /AV = (14 mg TAN /mmol) / (17 mg TA /mmol) = 14/17

AV

t 

## = (27.1652 mg TA /L) x (14 mg TAN / 17 mg TA )

= 22.3713 mg TA N/L

## 140

Step 5.

The AV in terms of TAN , or AV

t 

## , is converted from test pH 8.2 to pH 7 using the

equation for describing the pH

- dependence of acute values ( modified from Equation 11

in the 1999 AWQC document for normalization to pH 7 ) 2 

## :

( )

AV t, 7 

## = (AV

t 

## )/( 0.158673 ) = 140.990 mg N/L

Step 6. (temperature adjustment for invertebrates only)

The AV in terms of TAN at pH 7 , or AV t, 7 

## , is converted from this concentration at test

temperature to a standard test temperature of 2 0 ° 

## C using the equation shown below

( Equation 5 in the 1999 AWQC document ) 3 

## :

log(AV t, 7 ,2 0 

## ) = log(AV t, 7 

## )

- [

- 0.036(18 ° 

## C

- 2 0 ° 

## C)]

= 119.451 mg N/L

Because this is the only species in this genus for which data are in Table 1 in the 1984/1985

criteria document, 119.5 mg TA N/L is th e GMAV for the genus Dendrocoelum in Table 3 of this

update document.

2 

### The equation provided here should be applicable f rom pH 6 to 9, although uncertainty might exist at the lower end

of this range for certain species. Extrapolation below pH 6 is not advisable becaus e of the increasing scatter of the

data from the common regression line at lower pH, and extrapolation above pH 9 is not advisable because of

inadequate knowledge about the effect of the inhibition of ammonia excretion at high pH on results of toxicity tes ts

(Russo et al. 1988).

3 

### Note: Based on the 1999 reanalysis of the relationship between tem perature and ammonia toxicity, w hen test

temperature is less than 7°C, T should be set equal to 7, to reflect the plateau of the temperature

- toxicity relationship

a t these temperatures.

## 141

Appendix E. Conversion of Chronic Results of Toxicity Tests

.

As in the previous appendix with the acute results of toxicity tests, all of the ammonia

chronic values (EC 20 

## s) in Appendix B of this document were first converted to TAN at test

temperature and pH using the p K a relationship from Emerson et al. (1975). Once all the chronic

values were converted to total ammonia nitrogen, these values were then adjusted to pH 7 using

t he pH relationship developed in the 1999 AWQC document. After the adjustment to pH 7 , the

TAN chronic values were further normalized to a temperature of 2 0 ° 

## C for invertebrates only, as

per the recommendations in the 1999 AWQC document. The conversion pro cedure is illustrated

here using the data for the amphipod species Hyalella azteca.

Chronic value (CV) = EC 20 

## of 8.207 mg TAN/L

Test pH = 8. 04

Test Temperature = 25 .0 ° 

## C

Step s 1 through 4

.

( N ot required in this case as CV is already expressed in terms o f TAN

. F or more details

regarding these steps, see Appendix D ).

Step 5.

The CV in terms of TAN , or CV

t 

## , is converted from test pH 8. 04 to pH 7 using the

equation for describing the pH dependence of chronic values ( modified from Equation 12

in the 1999 AWQC document for normalization to pH 7 ) 4 

## :

( )

4 

### See footnote 3 in Appendix D.

## 142 CV t, 7 

## = (CV

t 

## )/( 0.38855 ) = 21.13 mg TA N/L

Step 6. ( T emperature adjustment for invertebrates only)

The CV in terms of TAN at pH 7 , or CV t, 7 

## , is conver ted from this concentration at test

temperature to a standard test temperature of 2 0 ° C using the equation shown below ( Equation 5

in the 1999 AWQC document ) 5 

## :

log(CV t, 7 ,2 0 

## ) = log(CV t, 7 

## )

- [

- 0.028( 25 ° 

## C

- 2 0 ° 

## C)]

= 29.17 mg TA N/L

Because this is the only species in this genus for which data in appendix B are available

,

29.17 mg TAN/L is the GMCV for the genus Hyalella reported in Table 4 of this update

document.

5 

### See footnote 4 in Appendix D.

## 143

Appendix F. Acute

- C hronic Ratios (ACRs)

.

The CCC was calculated directly from chronic value s (EC 20 

## s ) in Appendix B using the

standard fifth percentile procedure provided in the 1985 Guidelines (Stephan et al. 1985). As a

result, acute

- chronic ratios (ACRs) are not necessary for the derivation of the new chronic

criterion presented in this docu ment. It is still worthwhile, however, for EPA to provide

recommended ACRs for predicting chronic sensitivity of untested species using measured or

estimated acute values for other related efforts ( e.g. , developing Biological Evaluations in

support of Nat ional Endangered Species Act Consultations on EPA 304(a) criteria

recommendations, or when an ACR(s) is allowed to derive site

- specific criteria for ammonia in

fresh water). Table F.1 below presents ACRs for all species with chronic values that were used

in the derivation of a GMCV and for which comparable acute values were found, as well as for a

few additional species of special interest, such as threatened and endangered species. All acute

and chronic values were adjusted to pH 7 and to 20 ° 

## C (in the cas e of invertebrates). For each

species or genera where more than a single ACR was calculated, Species and Genus Mean

Acute

- Chronic Ratios (SMACRs and GMACRs, respectively) were also calculated as the

geometric mean value of individual ACRs and SMACRs. (No te: in the case of a single ACR

within a Genus, the ACR is the SMACR.) Additionally, taxon

- specific ACRs (TSACRs) were

calculated where practical and for purpose of comparison at the taxonomic level of Family and

Class.

The ACRs for freshwater aquatic i nvertebrates range from 2.406 to 49.45 (a factor of 21;

see Table F.1). Likewise, the ACRs for fish range from 3.437 to 36.53 (factor of 1 1 ). The broad

range in values can probably be explained because of the different kinds of chronic tests (life

cycle, ELS, 28

- d juvenile mussel or snail) and toxicological endpoints (survival, growth, or

reproduction) upon which they are based. The ACR of 36.53 for fathead minnow, for example,

was based on hatchability from the life

- cycle test of Thurston et al. (1986)

, whereas the early

life

- stage tests with fathead minnow of Mayes et al. (1986) and Swigert and Spacie (1983) gave

ACRs of 11.35 and 17.17. The range of ACRs based on chronic values from the two early life

stage tests is small, and it is perhaps not surpri sing that a life

- cycle test gave a higher ACR than

the early life

- stage tests. As another example illustrating the variability among ACRs from

different kinds of tests and using different toxicological endpoints, but this time comparing

## 144

amongst different species of invertebrates, the ACR of 49.45 for Lampsilis fasciola was based on

survival from a 28

- day test involving two month

- old juveniles (Wang et al. 2007a,b), whereas

the life

- cycle tests with the two species of cladocerans ( Ceriodaphnia acanthina and C. dubia )

are based on adverse effects on reproduction with ACRs of 2.406 (Mount 1982) and 3.924

(Nimmo et al. 1989), respectively (Table F.1).

The ACRs for bivalve mollusks in general are larger compared to other freshwater

aquatic animal taxa and range from 9.028 to 49.45. The ACRs for other freshwater invertebrates

range from 2.406 to 15.81. The ACRs for fishes, in contrast, are quite varied even within species

or genera. For example, the ACRs for Lepom is sp. range from 3.437 to 28.51 despite having

been based on ELS tests and using biomass or survival as the toxicological endpoint.

Figure F.1 depicts SMACRs in relation to SMAVs to determine whether there is a trend.

Only the weak trend of decreasing SM ACR with increasing SMAV is apparent; primarily due to

the comparatively large SMACRs for freshwater bivalve mollusks.

In general TSACRs for most freshwater aquatic animals (excluding bivalve mollusks) are

within the relatively small range of 5.113 to 15 .81 at the Class level, and may be acceptable for

use when certain taxon

- specific chronic toxicity data are not available. Perhaps not surprisingly,

the CCC (2.1 mg TAN/L) calculated as the quotient of the FAV of 32.99 mg TAN/L (at pH 7

and 20 ° C) and geom etric mean ACR for the Family Unionidae (15.52) agrees well with the CCC

calculated directly from available chronic data (see Appendix B and Figure 4).

## 145

Table F.1. Species, Genus and Taxon

- S pecific ACRs for Freshwater Aquatic Animals Exposed to Ammonia

.

Table F.1. Species, Genus and Taxon

- specific ACRs for Freshwater Aquatic Animals Exposed to Ammonia

Species Scientific Name Acute and Chronic

Test Endpoint pH Temp Normalized

Values Reference ACR SMACR GMACR TSACR

(Family) TSACR

(Class)

Class Gastropoda (Family: Lithoglyphidae )

Fluminicola sp. LC50 8.25 20.2 > 62.15

Besser 2011 7.940 7.940 7.940 7.940 7.940

EC20

- Change in Length 8.22 20.1 7.828

Class Bivalvia (Families Unionidae and Pasidiidae)

Lampsilis fasciola EC50 8.50 20.0 69.63 Wang et al. 2007b 49.45 49.45

21.13

15.52

25.68 E C20

- Survival 8.20 20.0 1. 408 Wang et al. 2007a

Lampsilis siliquoidea EC50 8.20 20.0 28.99 Wang et al. 2007a

9.028 9.028

E C20

- Su rvival 8.2 5 20.0 3.211 Wang et al. 2011

Vil losa iris EC50 8.40 20.0 23.29 Wang et al. 2007b

11.40 11.40 11.40 EC50 8.30 20.0 68.40 Wang et al. 2007b

E C20

- Survival 8.20 20.0 3.501 Wang et al. 2007a

Musculium transversum EC50 8.10 14.6 109.0 West 1985; Arthur et al. 1987

42.50 42.50 42.50 42.50

EC 20

- Survival 7.80 21.8 2.565 Sparks and Sandusky 1981

Class Branchiopda (Family: Daphniidae)

Ceriodaphnia acanthina EC50 7.06 24.0 154.3

Mount 1982 2.406 2.406

3.073

5.113 5.113 EC20

- Reproduction 7.15 24.5 64.10

Ceriodaphnia dubia EC50 7.80 25.0 152.9 Nimmo et al. 1989 3.924 3.924

EC20

- Reproduction 7.80 25.0 38.96

Daphnia magna EC50 8.50 20.0 296.9 Gersich and Hopkins 1986

8.186

8.507 8.507 EC20

- Reproduction 8.45 19.8 36.27 Gersich et al. 1985

EC50 8.34 19.7 419.1 Reinbold and Pescitelli 1982a 8.841

EC20

- Reproduction 7.92 20.1 47.40

Class Malacostraca ( Family: Dogielinotidae )

Hyalella azteca EC50 8.30 25.0 461.2 Ankley et al. 1995

15.81 15.81 15.81 15.81 15.81

EC20

- Biomass 8.04 25.0 29.17 Borgmann 1994

## 146

Table F.1. Species, Genus and Taxon

- specific ACRs for Freshwater Aquatic Animals Exposed to Ammonia

Species Scientific Name Acute and Chronic

Test Endpoint pH Temp Normalized

Values Reference ACR SMACR GMACR TSACR

(Family) TSACR

(Class)

Class Actinopterygii (Families Salmonidae, Catostomidae, Cyprinidae, Ictaluridae and Centrarchidae)

Oncorhynchus clarkii LC50 7.81 13.1 132.3 Thurston et al. 1978

5.122 5.122

5.518 5.518

8.973 O. clarkii henshawi EC20

- Survival 7.57 13.7 25.83 Koch et al. 1980

Oncorhynchus mykiss LC50 7.40 14.5 31.47 Calamari et al. 1981

9.696

5.945 EC20

- Survival 7.40 14.5 3.246 Calamari et al. 1977, 1981

LC50 7.67 7.7 40.40 Thurston et al. 1981a

3.646

EC20

- 5 yr Life Cycle 7.70 7.5

-

10.5 > 11.08 Thurston et al. 1984 a

Catostomus commersoni LC50 8.16 15.0 176.6 Reinbold and Pescitelli 1982c

14.75 14.75 14.75 14.75 LC50 8.1 4 15.4 166.3

EC20

- Biomass 8.32 18.6 11.62 Reinbold and Pescitelli 1982a

Notropis topeka LC50 8.09 13.2 147.3 Adelman et al. 2009

(EC 20 from Appendix C) 8.437 8.4 37 8.437

10.96 EC20

- Growth Rate 8.07 12.4 17.45

Pimephales promelas LC50 7.76 19.0 139.3

Thurston et al. 1983 , 1986 36.53

19.24 19.24 LC50 7. 83 22.0 158.7

LC50 7.91 18.9 178.9

LC50 7.94 19.1 162.3

LC50 8.06 22.0 205.0

LC50 8.03 22.1 216.3

EC20

- LC Hatchability 8.00 24.2 4.784

LC50 8.14 22.0 141.2 Mayes et al. 1986 11.35

EC20

- Survival 8.00 24.8 12.43

LC50 7.78 25.9 117.3

Swigert and Spacie 1983 17.17 LC50 7.80 25.6 126.8

EC20

- Biomass 7.82 25.1 7.101

Cyprinus carpio LC50 7.72 28.0 133.9 Hasan and MacIntosh 1986

8.100 8.100 8.100

EC20

- Gr owth: Weight 7.85 23.0 16.53 Mallet and Sims 1994

Ictalurus punctatus LC50 7.80 25.7 97.67 Swigert and Spacie 1983 4.800 4.800 4.800 4.800

EC20

- Biomass 7.76 26.9 20.35

Lepomis cyanellus LC50 7.72 22.4 144.3 McCormick et al. 1984 12.18

6.468 13.58 13.59 EC20

- Biomass 7.90 22.0 11.85

LC50 8.28 26.2 62.07 Reinbold and Pescitelli 1982a 3.437

## 147

Table F.1. Species, Genus and Taxon

- specific ACRs for Freshwater Aquatic Animals Exposed to Ammonia

Species Scientific Name Acute and Chronic

Test Endpoint pH Temp Normalized

Values Reference ACR SMACR GMACR TSACR

(Family) TSACR

(Class)

EC20

- Survival 8.16 25.4 18.06

Lepomis macrochirus LC50 7.60 21.7 93.31 Smith et al. 1984 28.51 28.51

EC20

- Biomass 7.76 22.5 3.273

Micropterus dolomieui LC50 (pH 6.5) 6.53 22.3 269.2

Broderius et al. 1985 31.12

13.61 13.61 EC20 (pH 6.5)

- Biomass 6.60 22.3 8.650

LC50 (pH 7.0) 7.16 22.3 144.3

14.84 EC20 (pH 7.0)

- Biomass 7.25 22.3 9.726

LC50 (pH 7.5) 7.74 22.3 105.2

6.670 EC20 (pH 7.5)

- Biomass 7.83 22.3 15.77

LC50 (pH 8.5) 8.71 22.3 126.0

11.14 EC20 (pH 8.5)

- Biomass 8.68 22.3 11.31

## 148

Figure F.1. SMACRs by SMAV Rank.

## 149

Appendix G. Results of the Regression Analyses of New Chronic Data for Unionid

Mussels

.

This appendix provides the figures generated using EPA’s TRAP program that was used

to c alculate EC 20 

## s for the new chronic ammonia toxicity studies conducted with unionid mussels

.

In the figures that follow, circles denote measured responses and solid lines denote estimated

regr ession lines. The model

- estimated EC 2 0 

## values and corresponding 95% confidence limits are

provided with each figure, as well as the pH and water temperature at which the test was

conducted. Per the text on page 3 2 in Chronic Toxicity to Fresh w ater Aquatic Animals and as

discussed in greater detail on page 56 in Effects Characterization , EPA decided that while 28

-

day survival EC 20 

## s from these tests using juvenile freshwater mussels are acceptable for

derivation of a chronic aquatic life criterion for ammonia

, EC 20 

## s based on growth response s from

these tests are not. The decision not to use the growth data from these tests was based on the

uncertainty in the test methods for assessing th e growth endpoint and the need for additional

research “to optimize feeding conditions, to conduct longer

- term exposures (e.g., 90 d), and to

compare growth effect to potential reproductive effect in partial life

- cycle exposure” (Wang et al.

2011). A dditionally, the growth response during these tests show a high degree of variability,

and the test methods for assessing growth, based on substrate or water

- only exposures, are

currently being evaluated

– see Figure below depicting the growth response of juvenile

fatmucket in the 28

- day tests reported in Wang et al. (2011).

## 150

## 151

## 152

## 153

## 154

Appendix H. Detailed Descriptions of Select New Acute and Chronic Toxicity Test Data

Used for Criteria Derivation

.

Acute Toxicity Test Data

Venustaconcha ellipsiformis (ellipse)

As noted above, the ellipse test data was not directly used in the acute criterion

calc ulation, but the data is described here as additional evidence supporting the determined acute

criterion value. The GMAV for the ellipse is based on the 24

- hr EC 50 

## reported for an acute

toxicity test initiated with 2

- hr old glochidia of the species (Wang et al. 2007b). Glochidia were

tested under static conditions at pH 8.1 and 20 ° C. Survival of control animals after 24 hours was

90 percent. The estimated measure d EC 50 

## at test temperature and pH was 4.450 mg N/L, after

adjusting the reported nominal E C 50 

## by m ultiplying by a factor of 0.89 (i.e., measured total

ammonia concentrations were 89 percent of nominal concentrations for 24 hour glochidia

exposures). The G MAV for this spe cies is 23.12 mg TAN/L when adjusted to pH 7 and 20 ° C

(Appendix A), and represents the lowest in the acute dataset (Table 3). The acute criterion of 17

mg TAN /L is considered protective of this species because the GMAV/2, a value used to

e stimate an effect level un

- differentiable from controls ( Federal Register on May 18, 1978 (43

FR 21506

- 18 ) , i s approximately 12 mg TAN/L for the ellipse, which is close to the current

criterion value

, given the variability and uncertainty in such toxicity tests.

Utterbackia imbecillis (pondshell mussel)

The GMAV for pondshell mussel of 46.93 mg TAN/L is the sixth lowest in the acute

dataset (Table 3). Although this GMAV is not one of the four used in calculating the FAV, the

value is composed of individua l EC 50 

## values ranging from a comparatively low acute value of

17.91 to 100.5 mg TAN/L (expressed as TAN and normalized to pH 7 and 20 ° C, Appendix A).

This GMAV is based on several EC 50 

## s (numbering nine in total) from three different studies

(Wade et al. 1 992; Keller 2000; Black 2001). This particular GMAV is based on tests with

predominantly juvenile mussels of various ages, but also including a single test which employed

glochidia (Appendix A). The pH and test temperature for all nine tests was relative ly uniform

and ranged from 7.80 to 8.35 and 24.0 to 25.1 ° C, respectively. Control survival exceeded 90

percent in all tests regardless of life

- stage tested.

## 155

Fusconaia masoni (Atlantic pigtoe)

The GMAV for the Atlantic pigtoe represents the seventh lowes t in the acute dataset, and

lies just below the lowest GMAV for the most sensitive fish species, the mountain whitefish

(Table 3). This GMAV is based on the 6

- hr EC 50 

## reported for an acute toxicity test initiated with

2

- hr old glochidia of the species (Bla ck 2001). Glochidia were tested under static conditions at

pH 7.6 and 24.9 ° C. Survival of control animals after 6 hours was 93 percent, falling to 87

percent after 12 hours. The EC 50 

## at test temperature and pH was 15.90 mg TAN/L, or 47.40 mg

TAN/L when adjusted to pH 7 and 20 ° C (Appendix A).

Fluminicola sp. (pebblesnail)

The GMAV of 62.15 mg TAN/L for Fluminicola is the tenth most sensitive in the acute

dataset (Table 3). As part of the study to evaluate the chronic sensitivity of pebblesnails

(Gastropoda: Hydrobiidae) to ammonia via 28

- day water only toxicity tests (see additional

details below under Chronic Toxicity Test Data: 28

- day Tests with Juvenile and Adult

Pebblesnails ( F luminicola sp ecies ) , Besser (2011) reported survival of ‘large’ snails (i.e., mean

starting shell length of 1.81 mm) after 96 hours of exposure. No mortality was observed in

controls through the highest test concentration of 8.801 mg TAN/L where 32 of 40 snails (80

percent) survived. The mean pH and test temperature at this highest ammonia treatment level

were 8.25 and 20.2 ° C, respectively. Because only 20 percent mortality occurred at this test

concentration, the EC 50 

## at test temperature and pH is recor ded in this document as > 8.801 mg

TAN/L, or >62.15 mg TAN/L when adjusted to pH 7 and 20 ° C (Appendix A).

Pleurocera uncialis (pagoda hornsnail)

Another non

- pulmonate snail species (pagoda hornsnail) was determined to be nearly as

sensitive to ammonia as pebblesnail, the pagoda hornsnail, which was ranked 12 th 

## in acute

sensitivity. Goudreau et al. (1993) collected and acclimated (for six days) adult snails from

Clinch River, Virginia prior to conducting a static renewal bioassay to determine a 96

- hr LC 50



## for this species. The test was conducted in a walk

- in experimental chamber set to a temperature

of 22 ° C and using chlorine free laboratory dilution water at pH 8. 1 . Survival of adult snails in

the control treatment was 100 percent. The reported LC 50 

## at test temperature and pH was 11.18

## 156

mg TAN/L when expressed as total ammonia. The LC 50 

## normalized to pH=7 and 20 ° C is 68.54

mg TAN/L (Appendix A).

Deltistes luxatis (Lost River sucker)

The endangered Lost River sucker is a freshwater fish species endemic to the Klamath

Basin of northern California and southern Oregon (Appendix A). The acute toxicity of ammonia

was determined for larval and juvenile Lost River sucker as reported in Saiki et al. (1999).

Larval tests were initiated when fish reared from sp awned eggs were 35 days old, whereas the

juvenile tests were initiated after the fish reached 3

- 7 months old. All fish were exposed for 96

hours under flow

- through conditions at pH 8.0 and 20 ° C. The reported LC 50 

## s at test temperature

and pH were 10.35 an d 16.81 mg/L for larval and juvenile fish, expressed as total ammonia

nitrogen (Appendix A). The LC 50 

## s normalized to pH 7 and 20 ° C are 44.42 and 72.18 mg

TAN/L, respectively (Appendix A). The GMAV for Lost River sucker is calculated as the

geometric mean of the two normalized LC 50 

## s, or 56.62 mg TAN/L (Table 3). Lost River sucker

represents the ninth most sensitive genus in the acut e dataset, and second most sensitive fish

species (following mountain whitefish which was the most sensitive GMAV) and is expected to

be protected by the CMC of 17 mg TAN/L.

Chronic Toxicity Test Data

28

- day Tests with Juvenile and Adult Pebblesnails (Flu minicola sp ecies )

The summary for 28

- day tests recently conducted with Fluminicola sp. includes the

results from repeat test s performed by Besser et al. in 2009 and 2010, the details of the latter of

which are summarized in a memorandum to EPA in 2011 ( thi s study referred to in this document

as Besser 2011).

Test organisms used in the Besser et al. (2009) 28

- day survival tests with wild

- caught

(Snake River, Idaho) Fluminicola sp

. included mixed

- aged adult and young

- adult organisms

(from 6 to 12 months). Mixed

- age classes were used because the acclimation cultures produced

only approximately 200 neonates for testing that were collected over a period of about four

months. Despite the fact that snails in the control treatment exhibited 100 percent survival, while

snails exposed to the highest ammonia concentration (7.9 mg TA N/L) exhibited 0 percent

survival

, extreme variation between replicates at the highest test concentrations was observed

## 157

during the test, i.e., snails in replicates were either all alive o r all dead in the 1.7 and 3.6 mg

TA N/L treatments. Based on the mean survivals for Fluminicola sp. , the reported survival EC 20



## for the species was estimated to be 1.02 mg TA N/L at test temperature (20.8 ° C) and pH (8.26)

,

or 3.90 0 mg TA N/L when adjusted to pH 7 and 2 0 ° C (see Appendix C ). The EC 20 

## reported for

the test is not considered reliable, however, due to the variability in survival among replicates in

the 1.7 and 3.6 mg TA N/L test concentrations; therefore, this data was not used in the derivation

of the final ammonia CCC (i.e., the all

- or

- none response in the replicates of these two treatments,

which, when averaged and used as means instead of analyzing the replicates separately in the

regression, allows estimation of an EC 20 

## tha t would otherwise be incalculable because of the

variability between treatment replicates). Thus, the upper limit CV for the test is uncertain. The

value clearly is a concentration below 7.9 mg TA N/L (at test temperature and pH) , but the exact

concentrat ion could not be determined at the time.

In an attempt to further define the 28

- day ammonia survival effects threshold for

Fluminicola sp., pebblesnails cultured in the laboratory at the USGS Columbia Environmental

Research Center were tested in April 2010 via a similar 28

- day test protocol (see Besser 2011).

This 2010 test was conducted with two size classes of juvenile pebblesnails: small (mean shell

length of 1.34 mm at the start of the test) and large (mean starting shell length of 1.81 mm).

Both size groups were exposed in the same flow

- through exposure system consisting of five

ammonia concentrations (ranging from a nominal concentration of 0.5 to 8 mg N/L in 50 percent

dilution series), plus a control, with four replicates of ten snails per replicat e (or 40 small and 40

large snails per treatment). Mean measured TAN concentrations, pH, and temperature were

maintained very close to target values throughout the test (i.e., mean measured ammonia

concentrations were within 14 percent of nominal, mean tr eatment pH ranged from 8.18 to 8.26,

and mean treatment water temperature ranged from 20.1 to 20.2 ° C). Survival was measured

after 4 and 28 days. Survival of snails after 28 days in the small size group was lower overall

(75 percent in the control and 60

- 68 percent in the nominal 0.5 to 2 mg TA N/L test concentration

range) in relation to that of the large size group (93

- 100 percent in both the control and low

ammonia test concentration). For both size groups, snail survival differed among test

concentrat ions and was s ubstantially lower than controls in the two highest ammonia

concentrations (4.0 and 8.0 mg TA N/L nominal), however, due to the lower control survival of

the small size group (<80 percent), the data for this group is not used quantitatively in the

## 158

derivation of the final ammonia CCC and is instead presented in Appendix C as other chronic

data.

Because the survival of the large size group of snails was acceptable in controls and snail

length different among concentrations according to concentrat ion

- response

, change in length for

the large size group was analyzed further for inclusion in the derivation of the CCC. (Note:

a ttempts to model concentration

- response curves for survival in the large size group using TRAP

software were not as informativ e because partial mortality was limited to only one treatment

(i.e., 28

- day survival ranged from 98 to 100 percent in the nominal 0.5, 1 and 2 mg TA N/L test

concentrations, only 10 percent in the 4 mg TA N/L nominal test concentration, and zero percent

at t he highest nominal test concentration of 8 mg TA N/L). T he growth E C 20 

## for this freshwater

non

- pulmonate snail species calculated using EPA’s TRAP (threshold sigmoid model with full

convergence) is 2. 281 mg TA N/L at test pH (8.22) and temperature (20.1 ° C), or ; 7.828 mg

TA N/L after adjustment to pH 7 and 2 0 ° C (see Appendix B ). The TRAP output for this test is

provided below to support the use of the growth

- based EC 20 

## for this particular species and test

.

## 159

Chronic Toxicity Tests with Juvenile Hyalella azteca

Borgmann (1994) conducted four sets of experiments on H. azteca using different

dilution water types and life

- stages of test organisms. One set of experiments consisted of tests

that began with <1

- week

- old organisms, all of which utilized weekly r enewals and dechlorinated

tap water originating from Lake Ontario. O f the three tests , one lasted four week s and the other

two lasted 10 weeks, the latter of which produced data on both surviva l and reproduction, as

described in detail in the 1999 AWQC document (U.S. EPA 1999). At the time, the results of

the two 10

- week tests were deemed sufficiently similar such that the results were analyzed

together and subsequently used as the basis for the pH and temperature adjusted EC 20 

## of <1.45

mg TAN/L (at pH 8 and temperature 2 5 ° C ) reported in Table 5 of the 1999 AWQC document

(U.S. EPA 1999). Since then, however, EPA has re

- evaluated the results of the three tests in

light of the recent extensive research that has been undertaken to elucidate the specific w ater

## 160

ionic composition and feeding requirements necessary to ensure the health of this particular

freshwater aquatic test organism for use in long

- term toxicity testing. During the EPA’s re

evaluation of these tests, it was concluded that while the ionic composition of the water used for

tes ting (dechlorinated c ity tap water originating from Lake Ontario) was acceptable, the results

of the two 10

- week chronic test s should not be used for deriving AWQ C for the following

reasons:

 Low control survival observe d after 10 weeks of exposure (only 66.3%), possibly linked

to inadequate food a nd feeding level that was employed, particularly after the first four

weeks of testing;

 P oor control reproduction observed after 10 weeks of exposure; and

 The fact that the amm onia concentrations increased substantially in critical test

treatments (e.g., the 0.1 mM ammonia treatment) during the final 3 weeks of testing

(weeks 7

– 10).

However, four week data for these two tests, in combination with data from the third

four

- week test with the same life stage, were not affected by these limitations. The measured

total ammonia concentrations and mean pH (8.04) reported for the “Tap water (young)” tests in

Table 1 of Borgmann (1994) reflect the analytical measurements combined from all three tests

conducted with this life stage (i.e., <1 wk old H. azteca ) . Likewise, the pooled results for

survival (from Figure 1a) and wet weight (from Tabl e 4) reflect the observations (weekly for

survival and after four weeks for wet weight) from the three respective tests, and thus, represent

observations stemming from six test replicates per treatment when combined. Using these data

up through the first four weeks of exposure, as well as the water temperature of 25 ° C

(maintained via an incubator) at which all sets of experiments in the study were run, a 28

- day

EC 20 

## of 29.17 mg N/L (based on biomass and normalized to pH 7 and 20 ° C) was calculated for

H. az teca for the study (Appendix B). These data were deemed sufficient to derive an SMCV for

the species (as an upper limit), which is subsequently used here for chronic criterion

development. This decision was largely predicated on the fact that:

 The ion co mposition of the water used in this test was acceptable;

 The control survival for the test s up through the first four weeks was good (88.4%); and

## 161

 The feeding l evel during the first four weeks of t est ing was acceptable (as judged via the

growth performance of the test orga nisms during this timeframe

.

New Chronic Data for Non

- salmonid Fish Species

Cyprinus carpio (common carp)

Mallet and Sims (1994) conducted a 28

- day early life

- stage test starting with e ggs

approximately 6 hours post

- fertilization. Mean pH and temperature for the test were 7.85 and

23 ° C, respectively. The measured DO concentrations reported for the test ranged from 79 to 94

percent of saturation. Ammonia had no effect on hatching succe ss at the highest concentration

tested (19.6 mg TA N/L); although survival of the post

- hatch stages was significantly reduced at

this level compared to controls (average fry survival in the control treatment was 86 percent).

Growth of fry was the most sens itive endpoint, and mean fry wet weights were inhibited at

concentrations ≥10.4 mg TA N/L. Even though the number of larvae in each replicate vessel was

not made uniform on hatching, at least one vessel per concentration contained an equivalent

stocking de nsity (23 to 29 carp), so the mean wet weight of carp in the one selected replicate per

concentration was analyzed using regression analysis. The resulting EC 20 

## value was 8.360 mg

TA N/L at 23 ° C and pH 7.85, which i s calculated to be 16.53 mg TA N/L at pH 7 , with a GMCV

sensitivity rank of ten ( see Appendix B and Table 4 ).

Esox lucius (northern pike)

Harrahy et al. (2004) conducted a 52

- day early life

- stage test starting with newly

fertilized northern pike embryos. The mean dissolved oxygen concentration in test water ranged

from 8.7 to 9.1 mg/L during the test. There was no effect of ammonia on hatching success up to

62.7 mg TA N/L, and larval survival of control fish was 100 percent. A significant reduction in

larval survival and growth was observed at concentrations of total ammonia ≥30.4 and 15.1 mg

TA N/L, respectively, at pH 7.6 2 and 8.7 ° C. The estimated EC 20 

## value reported for biomass was

13.44 mg TA N/L, which , normalized to pH 7 to support criteria development in this document,

is 20.38 mg TA N/L (Appendix B). T he GM CV of 20.38 mg TA N/L for northern pike is

included in Table 4 as the GMCV ranked 11 th 

## in sensitivity

.

## 162

New Chronic Toxicity Data for Salmonid Species

Chronic values for two additional studies with Oncorhynchus species are included in this

AWQC document. Koch et al. (1980) exposed Lahontan cutthroat trout ( Oncorhynchus clark i i

henshawi ) for 103 days in an ELS test. The measured dissolved oxygen concentrations for the

entire study ranged from 7.0 to 8.9 mg/L, with an overall average of 7. 9 mg/L. Survival of

embryos in the control treatment was 80 percent, with approximately 95 percent surviving

through the fry stage, and 80 percent surviving as fingerlings up to day 94 of the test. There

were no successful hatches at exposure levels o f 148 mg TAN/L or higher and no significant

mortality at exposure levels below 32.9 mg TA N/L. Regression analysis of the survival data

using an arcsine transformation resulted in a calculated EC 20 

## value of 17.89 mg TA N/L at 13.7 ° C

and pH 7.57. The EC 20 

## value is 25.83 mg N/L when adjusted to pH 7 ( Appendix B ).

The recent results of a 90

- day ELS test using a wild strain of rainbow trout exposed to

ammonia were reported by Brinkman et al. (2009). The test was initiated with newly fertilized

embryos (<24 h ) exposed under flow

- through conditions through hatch (28 days), swim

- up (15

days) and early fry development (52 days) to five concentrations of total ammonia with a control.

Each treatment consisted for four replicates containing 20 embryos each (N = 100 embryos per

treatment). Mean pH and temperature of test water measured among treatments was 7.75 and

11.4 ° C, respectively. Hatch success and survival of sac fry were similar to controls for all

ammonia concentrations, resulting in an unadjusted NOEC of > 16.8 mg TAN/L. Survival,

growth and biomass of swim

- up fry were significantly reduced at 16.8 mg TAN/L compared to

controls, but unaffected at 7.44 mg N/L, resulting in a chronic value (MATC) of 11.2 mg

TAN/L. The EC 20 

## calculated for biomass using TRAP a nd normalized to pH 7 is 15.60 mg

TAN/L (Appendix B).

## 163

Appendix I. Qualitative Weight

- of

- Evidence Test Data

.

Additional 28

- day Toxicity Test data for Freshwater Mussels

As part of the same study summarized above in the Effects Analyses to Freshwater

Aquatic Organisms under Summaries of Studies Used in Chronic Criteri on Determination (page

3 4 ) , Wang et al. (2007a) also attempted to determine the effect of ammonia on growth of 2

-

month old juvenile rainbow mussel, fatmucket , and wavy

- rayed lampmussel. The 28

- day test s

w ere conducted following the same methods (see ASTM 2006). The mean length of juvenile

rainbow mussel and fatmucket exposed to the lowest ammonia concentration s tested w as reduced

by 13 and 12 percent compared to mean length of control an imals, respectively, but increased by

7 percent for the wavy

- rayed lampmussel

. There was no consistent effect of ammonia, however,

on either length at 28 days or change in length after 28 days for fatmucket and wavy

- rayed

lampmussel at test concentrations where survival was unaffected; only the 28

- day test with

rainbow mussel exhibited such a concentration

- response for length and change in length. For

the reasons explained above under the section referenced , the growth endpoint was not used from

these te sts to derive the chronic criterion, and instead, the reported I C 25 

## (inhibition concentration)

estimated for these tests are included in Appendix C. The reported growth I C 25 

## for juvenile

rainbow mussel, fatmucket, and wavy

- rayed lampmussel from their resp ective 28

- day tests were

0.73, 0.44, and 0.57 mg TAN/L at test pH of 8.2 and temperature 20 ° C. These values, when

adjusted to pH 7 and 2 0 ° C

, are 2.406, 1.450 and 1.878 mg TAN/L, respectively ( see Appendix

C ).

Additional 28

- day Toxicity test Data for Fres hwater Snails

Besser et al. (2009), in a USGS study report completed for EPA, conducted 28

- day flow

through survival and growth tests with five species of snails, including four gill

- bearing (non

pulmonate) species and an air

- breathing (pulmonate) species. All tests were conducted in ASTM

hard water (mean hardness and alkalinity of approximately 170 and 120 mg/L as CaCO 3 

## ,

respectively) with a pH range of 8.20

- 8.29 and a temperature range of 19

- 21 ° C during testing.

Total ammonia nitrogen (mg TA N/L) concentrations in tests were measured weekly with the

percent of nominal concentrations ranging from 83 to 101 percent. Test results were based upon

## 164

the mean of the measured concentrations. For all snail exposures, the effect of ammonia on

growth wa s not determined for test species that were of mixed ages at test initiation (as explained

further below); growth, however, was not as sensitive of an endpoint as survival for at least one

( Pyrgulopsis idahoensis ) of the two snail species ( Lymnaea stagnali s and P

. idahoensis ) where

both growth and survival were measured (see Appendix C ).

Fontigens aldrichi (Ozark springsnail)

As part of the original study described above, Besser et al. (2009) also determined the

effect of ammonia on survival of the non

- pul monate snail F. aldrichi . Because F. aldrichi did

not reproduce during culturing and acclimation, field

- collected organisms of “older” (adult)

mixed

- ages were used for ammonia exposures. F. aldrichi exposed to ammonia in the 28

- day

test exhibited approxi mately 94 percent survival at 0.45 mg TA N/L , but only 50 percent at 0.83

mg TA N/L. Similar to the 2009 adult pebblesnail study, the replicates associated with the latter

0.83 mg TA N/L treatment in particular were characterized by high variability

, and the refore

,

these data were not used quantitatively in the derivation of the final ammonia CCC. In addition,

field

- collected F. aldrichi did not reproduce in captivity and animals in the control group did not

grow during testing. The reported EC 20 

## for F. ald richi was 0.61 mg TA N/L, or 2.332 mg TA N/L

when adjusted to pH 7 .0 and 2 0 ° C

, and is presented as other chronic data in Appendix C

.

Pyrgulopsis idahoensis (Idaho springsnail)

Two separate 28

- day tests with the de

- listed (from the Federal threatened and end angered

species list) non

- pulmonate snail species, P. idahoensis, were conducted which included

exposing juvenile organisms that were 7

- 9 and 11

- 13 weeks post

- hatch (organisms in each

cohort tested as separate replicates in the same test; test identified as test #3 in the 2009 Besser et

al. report ) , as well as a cohort of mixed

- age adults for all subsequent tests (test identified as test

#5 in Besser et al. 2009 ). The older life stages were chosen for testing because of the high

control mortality demonstr ated in preliminary tests using 2

- 3 week post

- hatch P. idahoensis.

In the 28

- day test with juveniles

, snails in four of the five test concentrations exhibited

≤44.4 percent survival , whereas control survival was 100 percent ; the single except ion being the

snails in the middle test concentration of 1.8 mg TA N/L, which demonstrated only 62.5 percent

survival

. The survival EC 20 

## reported for the test was 0.48 mg TA N/L at 20.1 ° C and pH 8.25 , or

## 165

1.726 mg TAN/L when adjusted to pH 7 and 2 0 ° C , however, due to the poor concentration

response relationship exhibited in this test, this EC 20 

## is highly uncertain , and therefore, the data

are included in Appendix C as “other data” and a re not used in the derivation of the CCC

.

The 28

- day chronic test initiated with mixe d

- aged adult P. idahoensis (4 to 8 months of

age) , on the other hand, resulted in an EC 20 

## reported for the test of 3.24 mg TA N/L , or 12.39 mg

TAN/L when adjusted to pH 7 and 2 0 ° C ( Appendix C ). Comparison of the juvenile and adult P.

idahoensis survival results indicates that juveniles are possibly the more sensitive of the two life

stages; however, due to the unreliability of the juvenile data, specifically the irregular survival

concentration

- response relationship, such an assertion is uncerta in at this time and the CVs are

not used quantitatively in the derivation of the CCC.

Taylorconcha serpenticola (Bliss Rapids snail)

A non

- pulmonate snail species listed under the Endangered Species Act, Taylorconcha

serpenticola, was exposed to ammonia i n 28

- day flow

- through toxicity tests as described above.

Because T. serpenticola did not reproduce or grow well during culturing and acclimation, field

collected organisms of “older” (adult) mixed

- ages were used. Survival of snails in the control

treatme nt was 100 percent, whereas survival of snails exposed to concentrations up to 3.6 mg

TAN/L exceeded 80 percent. Survival of snails exposed to the highest concentration tested (7.9

mg TAN/L) was reduced to only 30 percent. The survival EC 20 

## reported for T. serpenticola in

the test was 3.42 mg TA N/L at 20.8 ° C and pH 8.26 , or 13.08 mg TA N/L at pH 7 .0 and 2 0 ° C , but

because these snails did not grow well preceding the test, the data are also considered “ other

data ” and placed in Appendix C.

Pleurocera canaliculata (silty hornsnail)

EPA sponsored a study (GLEC 2011) to independently confirm the results of the 28

- day

juvenile and adult tests performed by the USGS, Columbia, MO laboratory (i.e., Besser et al.

2009 and Besser 2011) with non

- pulmonate snails. The USGS test results indica ted that

specialized and Federally

- listed non

- pulmonate gill

- bearing snails, such as the Idaho springsnail,

Bliss Rapids snail and pebblesnail, are potentially: 1) sensitive to prolonged, 28

- day ammonia

exposure, and 2) as sensitive as ammonia

- sensitive fr eshwater unionid mussel species to such

exposure. The EPA

- sponsored study involved a 28

- day flow

- through toxicity test using a more

## 166

widely

- distributed non

- pulmonate snail species, P. canaliculata. Two other non

- pulmonate snail

species were also selected based on distribution and generalized habitat preference; however, P.

canaliculata was the only one of the three wild

- caught snail species that were successfully held

and maintained in the laboratory for subsequent testing. Following a protocol similar to that

used in the USGS studies, a 28

- day toxicity test of mature, mixed

- age P. canaliculata was

conducted . The test design consisted of five ammonia test concentrations (0.9, 1.9, 3.8, 7.5, and

15 mg TAN/L, nominal) and one control, with four replicate ch ambers containing six snails each

per test concentration (N=24 snails per treatment). Test concentrations were based on the results

of a 96

- hr range finding test with the species, which provided a 96

- hr EC 50 

## of 9.66 mg TA N/L,

or approximately 88 mg N/L at 2 0 ° C and pH 7 .0. The endpoint for the 28

- day toxicity test was

mortality or immobilization, measured daily, the results of which were used to calculate an EC 20



## (at pH 7 and 2 0 ° C) of 1.845 mg TA N/L ( Appendix C ). However, due to the high degree of

tempora l variability in the measured total ammonia concentrations, as well as the unequal

response amongst replicates at the 1.9 mg TA N/L nominal test concentration, these data were not

used quantitatively in the derivation of the final ammonia CCC; a 28

- day ammo nia survival

effect concentration of < 7.667 mg TA N/L was recommended as the CV for the species which

supports the recent findings for the pebblesnails (1.8 mm) which were re

- tested and reported to

EPA via Besser ( 2011 )

.

( Note : The calculated EC 20 

## values using TRAP for P. idahoensis

, F. aldrichi , and T.

serptenticola , and the recommended 28

- day ammonia survival effects concentration of < 7.667

mg TA N/L for P. canaliculata , are deemed representative of non

- pulmonate snail sensitivity in

general and a re included in Appendix C for the purpose of comparison. )

Lymnaea stagnalis (pulmonate pondsnail)

The effect of ammonia in a 28

- day test on survival and growth of a third freshwater snail

species, the air

- breathing L. stagnalis, was also reported in Bes ser et al. (2009)

. The tests with L.

stagnalis utilized organisms that were <1 week post

- hatch due to the abundance of young

produced during culturing. L. stagnalis exposed to ammonia in a 28

- day flow

- through test

exhibited a pproximately 98 percent survi val at the highest concentration tested ( 8.0 mg TA N/L )

.

Because of the apparent negligible effect of ammonia on growth (i.e., the magnitude of the

## 167

growth reduction was so small, 6 percent at 1.8 mg TA N/L and only 16 percent at 8 mg TA N/L),

only the CV of > 8.0 mg TA N/L (for survival and growth) is reported in this document for the

test, or > 28.76 mg TA N/L when adjusted to pH 7 and 2 0 ° C. Note: For the purposes of this

document, t he CV for this test species is included in Appendix C and was not used in the

d erivation of the CCC because of the uncertainty of this value (> 28.76 mg TA N/L) as an upper

limit SMCV for the species.

Chronic Toxicity Data for Other Salmonids

A few other chronic toxicity tests produced applicable data for salmonid species that

were excluded from Appendix B and subsequent SMCV and GMCV calculation because either

the exposure did not include the appropriate life stage for the species , or the tests did not meet

other general 1985 Guidelines requirements for use in calculating the CCC. These tests are

summarized below and shown in Appendix C

.

The effects of water temperature and ammonia on the swimming characteristics of brook

charr ( Salvelinus fontinalis ) were investigated by Tudorache et al. (2010). Juvenile brook charr

wer e exposed to four ammonia concentrations in de

- chlorinated tap water for 96 hours at pH

9.10 and 15°C. The following swimming characteristics were measured in a 4.5 m long raceway

following this exposure: gait transition speed, maximum swimming speed, tai l

- beat amplitude,

tail

- beat frequency, maximum acceleration of bursts, number of bursts, distance of bursts, and

total swimming distance. The most sensitive swimming parameters (maximum swimming speed

and maximum acceleration) had a reported LOEC of 0.776 5 mg TAN/L, or 10.86 mg TAN/L

when normalized to pH 7.

The effects of long

- term exposure of ammonia on the molecular response of Atlantic

salmon ( Salmo salar ) parr were investigated by Kolarevic et al. (2012). The juvenile fish were

exposed for 105 days t o three concentrations of total ammonia nitrogen (TAN) in a flow

- through

apparatus with two different feeding regimes: full and restricted. Average water temperature

during the exposure was 12.1°C with a pH of 6.84. There was no effect of ammonia exposur e on

survival, resulting in a NOEC of 32.29 mg N/L (highest concentration tested) in the full feeding

regime. When normalized to pH 7, the CV for this test is >30.64 mg TAN/L.

Beamish and Tandler (1990) exposed juvenile lake trout ( Salvelinus namaycush ) f or 60

days on two different diets and observed a significant reduction in rate of weight gain when total

## 168

ammonia was 6.44 mg TA N/L at pH 8.02 and temperature was 11.6 ° C. Food intake by fish was

initially decreased at this concentration of total ammonia, b ut was no different from controls by

the end of the test. The growth LOEC for the study, when adjusted to pH 7 , was calculated to be

16.10 mg TA N/L. Note: this test was not included in the calculation of the CCC because it was

not a true ELS having been initiated with juvenile fish.

Chronic Toxicity Data for Threatened and Endangered Fish Species

Meyer and Hansen (2002) conducted a 30

- day toxicity test with late

- stage larvae (0.059

g) of Lost River suckers ( Deltistes luxatus ) at pH 9. 43 . The exposure d uration and pH were

chosen to represent the period of combined elevated unionized ammonia concentrations and

elevated pH that occur during cyanobacterial blooms in surface waters of Upper Klamath Lake,

which have been shown to last for several weeks to a m onth. Survival decreased significantly at

1.23 and 2.27 mg TA N/L, whereas the highest NOEC for all endpoints (survival, growth, body

ions, and swimming performance) was 0.64 mg TA N/L. Most deaths in the 2.27 mg TA N/L

exposure occurred during the first th ree days of the test, while mortality of larvae in the 1.23 0

mg TA N/L treatment occurred gradually from days 2 to 24. The 29 percent average mortality in

the 0.64 mg TA N/L treatment was all due to an unexplained complete loss of one replicate

between days 5 and 7 of the exposure. Control survival was > 90 percent. The calculated LOEC

of 1.23 0 mg TA N/L total ammonia normalized to pH 7 corresponds to a value of 25.31 mg

TA N/L , substantially higher than the 2013 chronic criterion value ( Appendix C ).

Fairchild et al. (2005) conducted 28

- day toxicity tests with early life stages of Colorado

pikeminnow ( Ptychocheilus lucius ) and razorback sucker ( Xyrauchen texanus ), and compared

the results of those tests with a test using a surrogate fish species, the f athead minnow

( Pimephales promelas ). Tests were initiated 2 days after swim

- up when the larvae were feeding

exogenously (or at 8

- day post hatch for Colorado pikeminnow, 9

- day post hatch for razorback

sucker, and 4

- day post

- hatch for fathead minnow). Temp erature, pH and dissolved oxygen over

the 28

- day test period averaged 19.9 ° C, 8.24, and 7.4 mg/L (80 percent saturation) over the

course of the three studies. Control mortality was 7 percent (fathead minnows and Colorado

pikeminnow) or less (3 percent, ra zorback sucker) on day 28. Effect concentrations based on the

survival and growth endpoints of the fathead minnow and razorback sucker tests were not

different; however, growth was the more sensitive endpoint for the Colorado pikeminnow test.

## 169

The 28

- day growth LOEC for the Colorado pikeminnow was 8.60 mg N/L, or 29.75 mg TAN/L

at pH 7, substantially greater than the 2013 chronic criterion. The 28

- day survival LOEC for the

razorback s ucker was 13.25 mg TAN/L, or 46.58 mg TAN/L at pH 7 . Both endangered fi sh

species exhibited similar sensitivity to ammonia as t he fathead minnow (LOEC of 32.71 mg

TAN/L at pH=7 ; see Appendix C )

. The same can be said for the Lost River sucker, which

indicates that these particular endangered fish species will be protected by the CCC value

calculated in this 201 3 AWQC Update.

Finally, Adelman et al. (2009) conducted both acute and chronic toxicity tests with

ammonia on the endangered Topeka shiner ( Notropis topeka ) and compared those values to

chronic studies with fathead minno ws. All tests used a flow

- through dosing apparatus and deep

well water with a total hardness and alkalinity of 210

- 230 mg/L CaCO 3 

## , and chloride

concentration of 0.64

- 1.04 mg/L. Acute survival studies with Topeka shiner lasted 96 hours and

were conducted o n two different life

- stages (juvenile and adult) and at two test temperatures,

warm, 25°C (adult and juvenile), and cold, 13°C (juvenile only). LC 50 

## s for total ammonia

ranged from 18.7

- 21.4 mg TAN/L at 25°C and 28.9 mg TAN/L at 13°C; all acute studies wer e

conducted at approximately pH 8. Normalized to pH 7, the 96

- hr LC 50 

## s were 69.59

– 88.27 mg

TAN/L at 25°C and 147.3 mg TAN/L at 13°C, both substantially greater than the acute criterion

value of 17 mg TAN/L, respectively (see Appendix A).

Chronic studies with Topeka shiners started with both adults and juveniles, since

embryos were not available, and lasted 30 days. The results of the survival and growth studies

with juvenile Topeka shiners were compared to a 30

- day juvenile survival study and 32

- day

embryo

- larval study conducted with fathead minnows in the same dilution water. The authors

interpreted the results of the relationship between the comparative studies using Topeka shiners

versus fathead minnows to infer what an expected result for an embryo

- larval study with Topeka

shiner would be. Reported MATC values (normalized to pH 8, according to USEPA 1999) were

16.95 mg TAN/L for the 30

- day juvenile fathead growth test and 8.62 mg TAN/L for the 32

- day

embryo

- larval survival and growth test. Using the relationship from the results obtained between

juvenile Topeka shiners and juvenile (growth) and embryo

- larval test using fathead minnows

(growth and survival), a 32

- day embryo

- larval study with Topeka shiner might be expected to

result in a chronic value that is approximately 51% more sensitive than the 30

- day juvenile

growth test with that species, or a chronic value of approximately 5.63 mg TAN/L (i.e., the

## 170

reported 30

- day MATC of 11.10 mg TAN/L at pH 8 based on growth of juvenile Topeka shiners

multiplied by a factor of 0.507). Using EPA’s TRAP (version 1.21a) the 32

- day biomass EC 20



## for embryo

- larval fathead minnow (measured from days 7

- 32), 30

- day adult survival EC 20 

## for

Topeka shiner, and 30

- day juvenile specific growth rate EC 20 

## for Topeka shiner were 7.457,

10.85, and 6.483 mg TAN/L at test temperatures (25.5, 23.9, and 12.4°C) and pH (7.95, 7.94,

and 8.07), respectively. When adjusted to pH 7, the EC 20 

## s for the respective tests are 16.87 mg

TAN/L for the fathead minnow (Appendix B ), and 24.21 and 17.45 mg TAN/L for the Topeka

shiner (Appendix C), much higher than the 2013 chronic criterion.

Chronic Toxicity Data for Amphibians

In a long term chronic study by Jofre and Karasov (1999), pre

- metamorphic (Gosner

stage 24

- 26) green frog ( Rana clamitans ) tadpoles were exposed to ammonia for 103 days under

renewal conditions. Tadpoles were evaluated in two different experiments conducted in

successive years. In the 1997 (repeat) experiment, survival and growth were not statistically

diff erent from controls at the highest concentration tested, or 2.2 mg TA N/L at pH 8.7 and 24 ° C,

although only approximately 50 percent of the frogs survived at this concentration compared to

the controls (98 percent survival). S urvival was reduced to approxi mately 78 percent at 0.94 16

mg TA N/L at test temperature and pH (or 7.149 mg TAN/L at pH 7) . Growth, measured as total

length, was no different between treatments. The frogs grew from an average total length of

approximately 7.5 mm at test initiation to approximately 50 mm in all treatments. The NOEC

for growth of green frog tadpoles in the study (which does not re flect an ELS or partial life cycle

test ) is > 16.74 mg TA N/L at pH 7

.

## 171

Appendix J

. Unused Acute Studies Potentially Influential for Freshwater Ammonia Criteria Development.

Appendix J. Unused Acute Studies Potentially Influential for Freshwater Ammonia Criteria Development

Reference: Organism: Reported or Normalized Acute

Value Expressed as Total

Ammonia (mg TA N/L) at pH= 7

and 2 0 °C, Where Applicable Rationale for Omission:

Abdalla, A.A.F. and C.D. McNabb. 1999. Acute and

sublethal growth effects of unionized ammonia to Nile

tilapia Oreochromis niloticus . In: Nitrogen production

and excretion in fish. Randall D.J. and D.D. Mackinlay

(Eds.), Department of Fisheries and Oceans, Vancouver,

BC, Canada and Towson University, Baltimore, MD.

pp. 35

- 48. Oreochromis niloticus Normalized LC 50 = 87.0 Species is a resident, non

- North American

"invasive" species known to cause or likely to cause

economic or enviro nmental harm (see ISAC 2006).

Because the species is in the Family Centrarchidae

which is well represented in the current acut e

criteria dataset, it has been intentionally excluded

from further consideration and calculation of a n

acute criterion.

Alonso, A. and J.A. Camargo. 2011. The freshwater

planarian Polycelis felina as a sensitive species to assess

the long

- term toxicity of ammonia. Chemosphere 84:

533

- 537. Polycelis felina Normalized 96 h LC 50 = 25.72 Species not resident in North America.

Ankley, G.T., M.K. Schubauer

- Berigan and P.D.

Monson. 1995. Influence of pH and hardness on toxicity

of ammonia to the amphipod Hyalella azteca . Can. J.

Fish. Aquat. Sci. 52(10): 2078

- 2083. Hyalella azteca Normalized 96 h LC 50 s:

Softwater (Lake Superior)

- 25.51 (pH 6.50)

47.35 (pH 7.49)

233.4 (pH 8.21)

Hardwater (Reconstituted

- ASTM)

- 232.8 (pH 6.55)

337.6 (pH 7.41)

545.5 (pH 8.45) Ankley et al. conducted several static

- r enewal acute

tests with H. azteca t o determine the effect o f pH

and hardness on the toxicity of ammoni a. For the

hardness evaluation, Ankley chose three waters for

testing, soft water (SW; unaltered lake Superior

water), moderately hard water (MW; hardened Lake

Superior water), and hard water (HW; hard

reconstitu ted water). At the time, Ankley et al.

focused only o n hardness in the test waters, but the

ion ratios in these three waters were not consistent.

Of the three water types, only t he moderately hard

water (MW) that Ankley used is suitable for testing

and c ulturing amphipods (see Appendix A for

results)

. The SW was not suitable for testing this

species because the sodium concentration was too

low. Similarly, the reconstituted HW was not

suitable because the bromide was too low. Bold

values indicate LC 50 s b elow the cutoff of 93 mg

TAN/L for unused, potentially influential acute

values.

## 172

Appendix J. Unused Acute Studies Potentially Influential for Freshwater Ammonia Criteria Development

Reference: Organism: Reported or Normalized Acute

Value Expressed as Total

Ammonia (mg TA N/L) at pH= 7

and 2 0 °C, Where Applicable Rationale for Omission:

Augspurger, T., A.E. Keller, M.C. Black, W.G. Cope

and F.J. Dwyer . 2003 . Water quality guidance for

protection of freshwater mussels (Unionidae) from

ammonia exposure. Envir on. Toxicol. Chem. 22(11):

2569

- 2575. Medionidus conradicus Normalized 48 h LC 50 = 27.56 48

- hr glochidia test. Secondary data from Keller

2000

Babu, T.R., P. Surendranath and K.V. Ramana Rao.

1987. Comparative evaluation of DDT and fenvalerate

toxicity on Penaeus indicus (H. Milne Edwards).

Mahasagar 20(4): 249

- 253. Daphnia magna Reported LC 50 s:

60 (25 h),

32 (50 h),

20 (100 h) pH not reported

– LC 50 s could not be normalized.

Belanger, S.E., D.S. Cherry, J.L. Farris, K.G.

Sappington and J.J. Cairns. 1991. Sensitivity of the

Asiatic clam to various biocidal control agents. J. Am.

Water Works Assoc. 83(10): 79

- 87. Corbicula fluminea Normalized LC 50 s:

23.55 (4.1 d)

64.99 (4.2 d) Species is a resident, non

- North American

"invasive" species known to cause or likely to cause

economic or environmental harm (see ISAC 2006).

This species is the target of current eradication and

control programs in va rious states, and because this

Phylum (Mollusca) is well represented in the current

acute criteria dataset, this species has been

intentionally excluded from further consideration

and calculation of an acute criterion.

Dehedin, A. , C. Piscart and P. Marmo nier. 2012

. Seasonal v ariations of the e ffect of t emperature on l ethal

and s ublethal t oxicities of a mmonia for three c ommon

f reshwater s hredders

. Chemopshere In press

. Gammarus pulex Normalized 96h LC 50 s:

36.98

49.31

49.31

69.40 Species not resident in North America. Control

mortality less than 15%

.

Gammarus roeselii Normalized 96 h LC 50 s:

2. 466

2 4 .66

36.98

46.27

46.27

55.31

55.31

57.84 Species not resident in North America. Control

mortality less than 15%

.

## 173

Appendix J. Unused Acute Studies Potentially Influential for Freshwater Ammonia Criteria Development

Reference: Organism: Reported or Normalized Acute

Value Expressed as Total

Ammonia (mg TA N/L) at pH= 7

and 2 0 °C, Where Applicable Rationale for Omission:

Dowden, B.F. and H.J. Bennett. 1965. Toxicity of

selected chemicals to certain animals. J. Water Pollut.

Control Fed. 37(9): 1308

- 1316. Daphnia magna Reported LC 50 s:

202 (24 h),

423 (25 h),

161 (48 h),

433 (50 h),

67 (72 h),

50 (96 h),

202, 139 (100 h) pH not reported

– LC 50 s could not be normalized.

Lymnaea sp. Reported LC 50 s:

241 (24 h),

173 (48 h),

73 (72 h),

70 (96 h) pH not reported

– LC 50 s could not be normalized.

Ewell, W.S., J.W. Gorsuch, R.O. Kringle, K.A.

Robillard and R.C. Spiegel. 1986. Simultaneous

evaluation of the acute effects of chemicals on seven

aquatic species. Environ. Toxicol. Chem. 5(9): 831

- 840. Daphnia magna Reported LC 50 in paper = >100;

Reported LC 50 in ECOTOX = >20

Normalized LC 50 = 36.29 Insufficient controls; pH that varied from 6.5

- 8.5

during t he exp osure. LC 50 based on a 96 h (non

standard) test duration.

Fairchild, J.F., A. Allert

, J. Mizzi, R. Reisenburg and B.

Waddell. 1999 . Determination of a safe level of

ammonia that is protective of juvenile Colorado

pikeminnow in the upper Colorado River, Utah. Final

Report.1998 Quick Response Program. U.S. Fish and

Wildlife Service, Region 2 (Salt Lake City Office). Pimephales promelas Normalized LC 50 = 60.12 72

- hour test in well water

Hazel, R.H., C.E. Burkhead and D.G. Huggins. 1982.

Development of water quality criteria for ammonia and

total residual chlorine for the protection of aquatic life in

two Johnson County, Kansas Streams. In: J.G. Pearson,

R.B. Foster, and W.E. Bishop (Eds.), Proc. Annu. Symp.

Aq. Tox., ASTM STP 766, Philadelphia, PA: 381

- 388. Etheostoma spectabile Normalized 96 h LC 50 s = 83.74,

71.12 Same data as in Hazel (1979)

– see E. spectabile in

Appendix A

.

Hecnar, S.J. 1995. Acute and chronic toxicity of

ammonium nitrate fertilizer to amphibians from

Southern Ontario. Environ. Toxicol. Chem. 14(12):

2131

- 2137. Pseudacris triseriata Reported values:

4

- d LC 50 = 17

4

- d NOEC = 5,

4

- d LOEC = 45 Formulation

- ammonium nitrate fertilizer

## 174

Appendix J. Unused Acute Studies Potentially Influential for Freshwater Ammonia Criteria Development

Reference: Organism: Reported or Normalized Acute

Value Expressed as Total

Ammonia (mg TA N/L) at pH= 7

and 2 0 °C, Where Applicable Rationale for Omission:

Hickey, C.W. and M.L. Vickers. 1994. Toxicity of

ammonia to nine native New Zealand freshwater

invertebrate species. Arch. Environ. Contam. Toxicol.

26(3): 292

- 298. Potamopyrgus

antipodarum Normalized 96 h LC 50 s:

33.14

29.79

38.93

36.27 Species is a resident, non

- North American

"invasive" species known to cause or likely to cause

economic or environmental harm (see ISAC 2006).

This species is the target of current eradication and

control programs in various states, and because this

Phylum (Mollusca) is well represented in the current

acute criteria dataset, this species has been

intentionally excluded from further consideration

and calculation of an acute criterion.

Horne, F.R. and S. McIntosh. 1979. Factors influencing

distribution of mussels in the Blanco River of Central

Texas. Nautilus 94(4): 119

- 133. Cyrtonaias tampicoensis Normalized LC 50 = 26.75 LC 50 based on a 7

- d (non

- standard) test duration.

Toxolasma texasensis Normalized LC 50 = 26.75 LC 50 based on a 7

- d (non

- standard) test duration.

Corbicula manilensis Normalized LC 50 = 26.75 LC 50 based on a 7

- d (non

- standard) test duration.

Jofre, M.B., and W.H. Karasov . 1999. Direct e ffect of

a mmonia on t hree s pecies of North American a nuran

a mphibians

. Environ.Toxicol.Chem. 18(8): 1806

- 1812

. Bufo americanus Normalized 96 h LC 5 0 = 62.85 Non

- standard acute endpoint based on hatch

success/ deformity .UIA calculated using Thurston et

al. (1979) EPA

- 600/3

- 79

- 091 from measured values

Rana clamitans Normalized 96 h LC 50 = 40.80 Non

- standard acute endpoint based on hatch

success/ deformity .UIA calculated using Thurston et

al. (1979) EPA

- 600/3

- 79

- 091 from measured values

Jofre, M.B., M.L. Rosenshield and W.H. Karasov . 2000.

Effects of PCB 126 and ammonia, alone and in

combination, on gree n frog ( Rana clamitans ) and

leopard frog ( R. pipiens ) hatching success, development,

and metamorphosis. J. Iowa Acad. Sci. 107(3 ): 113

- 122. Rana clamitans Normalized 96 h LC 50 = 49.56 Non

- standard acute endpoint based on hatch

success/ deformity

. pH not reported; assume same as

Jofre and Karasov 1999.

Kaniewska

- Prus, M. 1982. The Effect of ammonia,

chlorine, and chloramine toxicity on the mortality of

Daphnia magna Straus. Pol. Arch. Hydrobiol. 29(3/4):

607

- 624. Daphnia magna Normalized LC 50 = 1.980 LC 50 based on a 24

- h (non

- standard) test duration.

Me yer, J.S. and J.A. Hansen. 2002. Subchronic toxicity

of low dissolved oxygen concentrations, elevated pH,

and elevated ammonia concentrations to Lost River

suckers. Trans. Amer. Fish. Soc. 131: 656

- 666. Deltistes luxatus Normalized 48 h LC 50 :

78.23 The pH for this test was reported as 9.5, which is

outside of the acceptable pH range of (6.0

- 9.0) these

criteria were meant to apply.

Morgan, W.S.G. 1979. Fish l ocomotor b ehavior p atterns

as a m onitoring t ool. J. Water Pollut. Control. Fed.

51(3): 580

- 589. Micropterus salmoides Normalized EC 50 = 5.010 Acute toxicity evaluated electronically based on

activity. Ex posure was only 24

- h (non

- standard) in

test duration. Concentrations were nominal.

Morgan, W.S.G. 1976. Fishing for toxicity: Biological

automonit or for continuous water quality control. Effl.

Water Treat. J. 16(9): 471

- 475. Micropterus salmoides Normalized EC 50 = 5.010 Added nominal concentrations equivalent to 48

- h

LC 50 from previous literature values, then monitored

opercular rhythm activity for 24 h.

## 175

Appendix J. Unused Acute Studies Potentially Influential for Freshwater Ammonia Criteria Development

Reference: Organism: Reported or Normalized Acute

Value Expressed as Total

Ammonia (mg TA N/L) at pH= 7

and 2 0 °C, Where Applicable Rationale for Omission:

Morgan, W.S.G. and P.C. Kuhn. 1974. A method to

monitor the effects of toxicants upon breathing rate of

largemouth bass ( Micropterus salmoides Lacepede).

Water Res. 8(1): 67

- 77 Micropterus salmoides

Lacepede Normalized EC 50 s:

110.3 (11 h),

31.32 (22 h),

110.3 (23 h),

1.556 (44 h) Similar to Morgan (1976). This is not an actual

toxicity test. Rather, it is a test of a monitoring

system that relates nominal LC 50 concentrations

(based on literature values), to breathing rate

monitored over 24 h.

Morgan, W.S.G. 1978. The use of fish as a biol ogical

sensor for tox ic comp arison in potable water. Prog.

Water Tech. 10: 395

- 398. Micropterus salmoides Normalized LC 50 = 9.091 Similar to other Morgan studies listed in this table

where n ominal ammonia concentrations based on

literature LC 50 concentrations are added to tanks and

breathing rate and activity level are monitored

electronically for 24 h.

Passell, H.D., C.N. Da hm and E.J. Bedrick. 2007.

Ammonia modeling for assessing potential toxicity to

fish species in the Rio Grande, 1989

- 2002. Ecol. Appl.

17(7): 2087

- 2099. Hybognathus amarus Secondary data; reported LC 50 from

Buhl 2002 = 1.01 mg/L unionized

ammonia

- N In this study the fr equency of acute ammonia

exceeda nces were modeled by relating discharge,

pH, temperature, and stream ammonia

concentrations to literature LC 50 values.

Scheller, J.L. 1997. The effect of dieoffs of Asian clams

( Corbicula fluminea ) on native fr eshwater mussels

( U nionidae). Virginia Polytechnic Institute and State

University, Blacksburg, VA. Corbicula fluminea Normalized LC 50 s:

6.498 (96 h)

11.57 (96 h)

14.62 (96 h) Species is a resident, non

- North American

"invasive" species known to cause or likely to cause

economic or environmental harm (see ISAC 2006).

This species is the target of current eradication and

control programs in various states, and because this

Phylu m (Mollusca) is well represented in the current

acute criteria dataset, this species has been

intentionally excluded from further consideration

and calculation of an acute criterion.

Pimephales promelas Normalized LC 50 = 38.46 Non

- standard (48 h) test du ration.

Watton, A.J. and H.A. Hawkes. 1984. The acute toxicity

of ammonia and copper to the gastropod Potamopyrgus

jenkinsi (Smith). Environ. Pollut. Ser. A 36: 17

- 29. Potamopyrgus jenkinsi Normalized EC 50 s:

40.31 and 42.06 (48 h),

27.60 and 27.17 (96 h) Species not resident in North America.

## 176

Appendix J. Unused Acute Studies Potentially Influential for Freshwater Ammonia Criteria Development

Reference: Organism: Reported or Normalized Acute

Value Expressed as Total

Ammonia (mg TA N/L) at pH= 7

and 2 0 °C, Where Applicable Rationale for Omission:

Whiteman, F.W., G.T. Ankley, M.D. Kahl, D.M. Rau

and M.D. Balcer . 1996. Evaluation of interstitial w ater

as a route of exposure for ammonia in sediment tests

with benthic m acroinvertebrates

. Environ. Toxicol.

Chem. 15(5): 794

- 801

. Hyalella azteca Normalized 96 h LC 50 s:

10.27 (Lake Superior water)

11.06 (sediment test)

72.67 (sediment test) Tests were fed. The results from the two s ediment

test s were not used because sediment toxicity tests

using pore water measurements likely underestimate

the toxicity of ammonia in a water

- only exposure

, i.e., test animals could have been exposed to the

higher interstitial ammonia concentrations during

the exposure a . The 96 h LC 50 for H. azteca from

water

- only exposure to L ake Superior water was not

used from this study because the sodium

concentration in this dilution water is too low for

maintaining adequate animal health

– see also the

results in this appendix from Ankley et al. (1995)

above.

a For the same reason the sediment tests reported by Whiteman et al. (1996) for H. azteca were unused for criteria derivation, results from the sediment tests from Besser et al.

(1998) were also not used. The normalized 96 h LC 50 s for H. azteca from the Besser et al. (1998) sedime nt tests were 120.5 and 321.4 mg TAN/L at pH 6.69 and 7.56,

respectively. Two other LC 50 s generated for H. azteca which are also not used for criteria derivation (due to the insufficient amount of detail provided) include values of 251.5 and

262.7 mg TAN/ L from Sarda (1994). Because these latter values exceed 93 mg TAN/L, they are considered non

- influential data for the purpose of criteria derivation.

## 1 77

Appendix K

. Unused Chronic Studies Potentially Influential for Freshwater Ammonia Criteria Development.

Appendix K. Unused Chronic Studies Potentially Influential for Freshwater Ammonia Criteria Development

Reference: Organism: Reported or Normalized Chronic Value

Expressed as Total Ammonia (mg

TA N/L) at pH= 7 and 2 0 °C, Where

Applicable Rationale for Omission:

El

- Shafai, S.A., F.A. El

- Gohary, F.A. Nasr, N.P.

Vander Steen and H.J. Gijzen. 2004. Chronic

ammonia toxicity to duckweed

- fed tilapia

( Oreochromis niloticus ). Aquacult. 232(1

- 4): 117

-

127. Oreochromis

niloticus Normalized Chronic value = 6.881 (7 5 d) Test was a 35

- day juvenile test; not a true fish ELS

test. Species is also a resident, non

- North American

"invasive" species known to cause or likely to cause

economic or environmental harm (see ISAC 2006).

DeGraeve, G.M., W.D. Palmer, E.L. Moore, J.J.

Coyle and P.L. Markham. 1987. The effect of

temperature on the acute and chronic toxicity of

unionized ammonia to fathead minnows and channel

catfish. Battelle, Columbus, OH. Ictalurus punctatus Normalized 30

- day NOEC = 0.5628 Per the 1999 update, this 30

- day test with juvenile

catfish encountered some problems that precluded

effective use of the se data. For example, s ome of

the test organisms were treated with acriflavine up

to two days prior to the beginni ng of the test. In

addition, the mean measured DO concentration was

below 5.5 mg/L and below 6 0 percent of saturation

in some of the treatments.

Hecnar, S.J. 1995. Acute and chronic toxicity of

ammonium nitrate fertilizer to amphibians from

Southern Ontar io. Environ. Toxicol. Chem. 14(12):

2131

- 2137. Pseudacris

triseriata Reported values:

100

- d NOEC = 2.5,

100

- d LOEC = 10 Formulation

- ammonium nitrate fertilizer.

Hermanutz, R.O., S.F. Hedtke, J.W. Arthur, R.W.

Andrew and K.N. Allen. 1987. Ammonia effect s on

macroinvertebrates and fish in outdoor experimental

streams. Environ. Pollut. 47: 249

- 283. Ictalurus punctatus Normalized NOEC = 4.369 S urvival and growth of juvenile channel catfish

were evaluated via exposure to ammonia in

experime ntal streams. Three s eparate tests lasted

from 36 to 177 days and were started with

individuals whos e average weights ranged from 6

to 19 g. Average temperatures i n the three tests

were 17 to 21 ° C. Both of th e longer tests showed

monotonic, substantial reductions in biomass; these

results are in reasonable agreement with the results

of the laboratory tests. However, juveniles might

not be as sensitive to ammonia toxicity as early life

stages are. These results are not included because

they are from a field study where ammonia

concentrations were highly variable.

Sander vitreus Normalized NOEC = 4.182 Omitted for the same reasons as was Ictalurus

punctatus

.

## 178

Appendix K. Unused Chronic Studies Potentially Influential for Freshwater Ammonia Criteria Development

Reference: Organism: Reported or Normalized Chronic Value

Expressed as Total Ammonia (mg

TA N/L) at pH= 7 and 2 0 °C, Where

Applicable Rationale for Omission:

Hickey, C.W., L.A. Golding, M.I. Martin and G.F.

Croker. 1999. Chronic toxicity of ammonia to New

Zealand freshwater invertebrates: A meso co sm

study. Arch. Environ. Contam. Toxicol. 37:338

- 351. Deleatidium sp.

(Ephemeroptera) Normalized 29

- day EC 25 (survival) =

3.844 Species not resident in North America. These

results are not included because they are from a

field study where ammonia concentrations were

highly variable.

Rice, S.D. and J.E. Bailey. 1980. Survival, size, and

emergence of pink s almon, Oncorhynchus

gorbuscha , alevins after short

- and long

- term

exposures to ammonia. Fish. Bull. 78(3):641

- 64 8. Oncorhynchus

gorbuscha Normalized 61

- d NOEC = 5.859 Per the 1999 update, t he only chronic test began

sometime after hatch and ended when the alevins

emerged (i.e., at the beginning of swim

- up);

therefore the test did not include effects of

ammonia on the g rowth and survival of fry after

feeding started. In addition, no information was

given concerning survival to the end of the test in

the control or any other treatment. This test did not

provide data concerning survival and is not an ELS

te st because it be gan after hatch

. Schulter, M. and J. Groeneweg . 1985. The inhibition

by ammonia of population growth of the rotifer,

Brachionus rubens , in continuous culture.

Aquaculture 46: 215

- 220

. Brachionus rubens Normalized 7

- d NOEC = 3.000 Species is not resident i n North America . Generally

a marine Rotifera. Undescribed culture medium.

NOEC based on population growth of cultures.

Smith, C.E. 1972. Effects of metabolic p roducts on

the quality of rainbow trout. Am. Fish. Trout News

17:7

- 8. Oncorhynchus

mykiss Normalized 84

- d NOEC = 2.304 This test did not provide data concerning survival

and is not an ELS te st because it began after hatch

. The authors reported that as long as the DO

concentration was maintained at 5 mg/L or greater,

growth of young rainbow tro ut was not

significantly redu ced until average total ammonia

concentrations reached 1.6 mg TAN /L at test pH

and temperature (7.75 and 10 ° C, respectively).

Zischke, J.A. and J.W. Arthur. 1987. Effects of

elevated ammonia levels on the fingernail clam,

Musculium transversum , in outdoor experimental

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transversum Normalized LOEC = 6.933 (survival) This was a flow

- through, measured mesocosm

experiment performed in the field. The test

concentrations varied during the length of the

experiment.

## 179

Appendix L. Unused (Non

- I nfluential) Acute and Chronic Studies for Freshwater

Ammonia Criteria Development

– Screened Out Studies with Code List

. (appears separately at end of appen dix)

Appendix L. Unused (Non

- influential) Acute and Chronic Studies for Freshwater Ammonia Criteria Development

Citation ECOTOX or

Other

Ref. No Rejection

Code(s) Comment(s)

Academy of Natural Sciences. 1960. The s ensitivity of

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Other

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Appendix L. Unused (Non

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Citation ECOTOX or

Other

Ref. No Rejection

Code(s) Comment(s)

Buikema, A.L.

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## 182

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## 191

Appendix L. Unused (Non

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Citation ECOTOX or

Other

Ref. No Rejection

Code(s) Comment(s)

Redner, B.D. and R.R. Stickney. 1979. Acclimation to

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Appendix L. Unused (Non

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Citation ECOTOX or

Other

Ref. No Rejection

Code(s) Comment(s)

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## 193

Appendix L. Unused (Non

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Citation ECOTOX or

Other

Ref. No Rejection

Code(s) Comment(s)

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## 194

Appendix L. Unused (Non

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Citation ECOTOX or

Other

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Code(s) Comment(s)

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## 195

Appendix L. Unused (Non

- influential) Acute and Chronic Studies for Freshwater Ammonia Criteria Development

Citation ECOTOX or

Other

Ref. No Rejection

Code(s) Comment(s)

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Other

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Code(s) Comment(s)

Woltering, D.M., J.L. Hedtke and L.J. Weber. 1978.

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## 197

Corresponding Code List

ABIOTIC FACTOR

(AF) Studies where one or both of the two abiotic factor s ( pH and temperature ) important for ammonia criteria

derivation are not reported

.

ACELLULAR

(Ace) Studies of acellular organisms (protozoa) and yeast.

BACTERIA

(Bact) Studies describing only the results on bacteria.

BIOMARKER

(Biom) Studies reporting results for a biomarker having no reported association with a biologically significant

adverse effect (survival, growth, or reproduction of an individual o r population) and an exposure dose (or

concentration).

CONTROL

(Con) Studies where control mortality is insufficient or unsatisfactory, i.e., where survival is less than 90% in

acute tests or 80% in chronic tests; or where no control is used.

DETAIL

(Det ) Insufficient detail regarding test methodology or statistical analysis.

DURATION

(Dur) Laboratory and field studies where duration of exposure is inappropriate (e.g., too short) for the type of

test (i.e., acute or chronic), or was not reported or could not be easily estimated.

EFFLUENT

(Efflu) Studies reporting only effects of effluent, sewage, or polluted runoff where individual pollutants are not

measured.

EFFECT

(Eff) Studies where the biologically significant adverse effect was not survival, grow th, or reproduction of an

individual or population.

ENDPOINT

(UEndp) Studies reported in ECOTOX where an endpoint (LC50, EC50, NOEC, LOEC, MATC, EC20, etc.) was

not provided, where none of the concentrations tested in a chronic test were deleterious (no LOEC); or

where all concentrations tested in a chronic test caused a statistically significant adverse effect (no

NOEC).

FIELD

(Field) Chronic, long

- term studies conducted in a field setting (stream segment, pond, etc.) where source/dilution

water is not characterized for other possible contaminants.

FORMULATION

(Form) Studies where the chemical is a primary ingredient in a commercial formulation, e.g., biocide, fertilizer,

etc.

INVASIVE [Harmful]

(INV) Defined in this document as a species that is non

- native to the ecosystem under consideration and

whose introduction causes or is likely to cause economic or environmental harm or harm to human health

(see ISAC 2006).

IN VITRO

(In Vit) In vitro studies, including only exposure of the chemical to cell cultures and excised tissues and not

related to whole organism toxicity.

LETHAL TIME

(LT) Laboratory studies reporting only lethal time to mortality, except under special conditions (no other

applicable information is available for species pivotal in mak ing a finding).

NO DOSE or CONC

(No Dose or Conc) Studies with too few concentrations to establish a dose

- response, or no usable dose or concentration

reported in either primary or sister article(s), except under special conditions (no other applicable

information is available for species pivotal in making a finding).

NOMINAL

(Nom) Chronic studies where test concentrations were not measured.

NON

- RESIDENT

(NonRes) Species that are not resident to North America, or where there is no reported evidence of their

reproducing naturally in North America.

NO ORGANISM

(No Org) Laboratory and field studies where no one organism is studied (e.g., periphyton community) or where no

scientific/common name is given in either a primary or sister article(s).

PURITY

(Pur) Studies where the chemical purity of the toxicant was less than 80% pure (active ingredient).

ROUTE OF EXPOSURE

(RouExp) Dietary or un

- natural exposure routes for aquatic chemicals, e.g., injection, spray, inhalation.

Secondary

(Sec) Non

- original data first reported elsewhere.

Sediment Exposure

(SedExp) Sediment

- based toxicity test and method.

TOXICANT

(Tox) Inappropriate form of toxicant used or none identified in a laboratory or field study. Note: Inappropriate

form includes mixtures.

UNACCEPTABLE CHRONIC

(UChron) Chronic studies which were not based on flow

- through exposures (exception for cladocerans and other

small, planktonic organisms where test water is continuously renewed)

, where test concentrations were

not measured , or when th e chronic test did not include the appropriate test duration for the organism and

life

- stage tested

.

## 198

UNUSUAL DILUTION

WATER

(Dilut) Laboratory or field studies where the dilution water contained unusual amounts or ratios of inorganic ions

or was without addition of appropriate salts (i.e., distilled or de

- ionized water).

VARIABLE EXPOSURE

(VarExp) Excessive variability in contaminant concentrations during the exposure period.

WATER QUALITY

(WatQual) Studies where the measured test pH is below 6 or greater than 9, where dissolved oxygen was less than

40% saturation for any length of time, or where total or dissolved organic carbon is greater than 5 mg/L.

## 199

Appendix M

. 1999 Re

- examination of Temperature Dependence of Ammonia Toxicity

.

This section presents the temperature analysis published in the 1998 Update, followed by

the re

- analysis performed for th e 1999 Update and reproduced here as background information

.

Figure and table numbers are preceded by an ‘M’ in this appendix, in order to distinguish them

from tables and figures in the main document.

1998 Analysis of Temperature

- Dependence

The 1984/1985 ammonia criteria document identified temperature as an important factor

affecting the toxicity of ammonia. When expressed in terms of un ionized ammonia, the acute

toxicity of ammonia was reported in the criteria document to be inversely related to temperature

for several species of fish, whereas limited data on acute ammonia toxicity to invertebrates

showed no significant temperature depen dence. No direct data were available concerning the

temperature dependence of chronic toxicity. It was noted, however, that the differences between

chronic values for salmonid fish species tested at low temperatures and chronic values for

warmwater fish species tested at higher temperatures paralleled differences in acute toxicity

known to be caused by temperature.

In the 1984/1985 criteria document, an average temperature relationship observed for fish

was used to adjust fish acute toxicity data to a c ommon temperature (20°C) for derivation of the

CMC for unionized ammonia; this same relationship was used to extrapolate this CMC to other

temperatures. (Invertebrate toxicity data were not adjusted, but invertebrates were sufficiently

resistant to ammoni a that adjustment of invertebrate data was not important in the derivation of

the CMC.) This temperature relationship for fish resulted in the unionized ammonia CMC being

higher at warm temperatures than at cold temperatures. Additionally, because of con cerns about

the validity of extrapolating the temperature relationship to high temperatures, the unionized

ammonia CMC was "capped" to be no higher than its value at a temperature, called TCAP, near

the upper end of the temperature range of the acute toxic ity data available for warmwater and

## 200

coldwater fishes. Similarly, the CCC was capped at a temperature near the upper end of the

temperature range of the available chronic toxicity data.

Although the unionized ammonia criterion is lower at low temperature s, this does not

result in more restrictive permit limits for ammonia because the ratio of ammonium ion to

unionized ammonia increases at low temperatures, resulting in the total ammonia criterion being

essentially constant at temperatures below TCAP. In practice, however, the criterion at low

temperatures can be more limiting for dischargers than the criterion at high temperatures because

biological treatment of ammonia is more difficult at low temperatures. Above TCAP, the

constant unionized ammonia cri terion results in the total ammonia criterion becoming

progressively lower with increasing temperature, which can also result in restrictive discharge

limitations.

Because more data are available at moderate temperatures than at lower and higher

temperatu res, the ammonia criterion is most uncertain for circumstances when compliance can

be most difficult, either because of the low total ammonia criterion at high temperatures or

because of treatment difficulties at low temperatures. This section examines th e data used in the

1984/1985 criteria document and newer data to determine (1) whether the use of TCAPs should

be continued and (2) whether a lower unionized criterion at low temperature is warranted. Data

used include those analyzed by Erickson (1985), w hich are shown in Figure 2 of the 1984/1985

document, and more recent data reported by Arthur et al. (1987), DeGraeve et al. (1987), Nimmo

et al. (1989), and Knoph (1992).

Data not used include those reported by the following:

1. Bianchini et al. (1996) cond ucted acute tests at 12 and 25°C, but one test was in fresh

water, whereas the other was in salt water.

2. Diamond et al. (1993) conducted acute and chronic toxicity tests on ammonia at 12 and

20°C using several vertebrate and invertebrate species. When expr essed in terms of

unionized ammonia, they reported that vertebrates (i.e., fishes and amphibians) were

more sensitive to ammonia at 12°C than at 20°C, whereas invertebrates were either less

## 201

sensitive or no more sensitive at 12°C, compatible with the relati onships used in the

1984/1985 criteria document. However, such factors as dilution water and test duration

varied between tests at different temperatures and possibly confounded the results (see

Appendix 1 of the 1999 update ), raising doubts about the tem perature comparisons for

the vertebrates and invertebrates.

Arthur et al. (1987) measured the acute toxicity of ammonia to several fish and

invertebrate species at ambient temperature during different seasons of the year. For three of the

five fish spec ies (rainbow trout, channel catfish, and white sucker), the relationship of toxicity to

temperature was similar to that used in the 1984/1985 criteria document. When expressed in

terms of unionized ammonia, no clear relationship existed between temperatur e and toxicity for

the other fish species (fathead minnow and walleye). This result for the fathead minnow is

different from those of three other studies ( Reinbold and Pescitelli 1982 b ; Thurston et al. 1983;

DeGraeve et al. 1987) reporting a significant e ffect of temperature on the acute toxicity of

unionized ammonia to the fathead minnow. For five invertebrate species, each tested over a

temperature range of at least 10°C, there was no consistent relationship between temperature and

unionized ammonia tox icity. An initial report of these results (West 1985) was the basis for no

temperature adjustment being used for invertebrate data in the 1984/1985 criteria document.

Further analysis of the Arthur (1987) data is discussed later.

DeGraeve et al. (1987) studied the effect of temperature (from 6 to 30°C) on the toxicity

of ammonia to juvenile fathead minnows and channel catfish using acute (4

- day) and chronic

(30

- day) ammonia exposures. As shown for both fish species in Figure M. 1, log(96

- hr unionized

amm onia LC 50 

## ) versus temperature was linear within the reported uncertainty in the LC 50 

## s; the

slopes were similar to those reported in the 1984/1985 criteria document. Problems with the

channel catfish chronic tests precluded effective use of those data and the highest tested

ammonia concentrations in the fathead minnow chronic tests at 15 and 20°C did not cause

sufficient mortality to be useful. However, sufficient mortality did occur in the fathead minnow

chronic tests at 6, 10, 25, and 30°C. Based on reg ression analysis of survival versus log

## 202

concentration (discussed in more detail in the section concerning the CCC below), 30

- day LC20s

for unionized ammonia were 0.11, 0.18, 0.48, and 0.44 mg N/L at 6, 10, 25, and 30°C,

respectively. This temperature depe ndence (Figure M

. 1) is similar to that for acute toxicity and

that used in the 1984/1985 criteria document. The actual effect of temperature on these 30

- day

LC20s is probably somewhat greater, because test pH decreased with increasing temperature.

Nimmo et al. (1989) conducted acute toxicity tests on ammonia at 6 and 20°C in a well

water using Johnny darters and in a river water using both Johnny darters and juvenile fathead

minnows. In all three sets of tests, LC 50 

## s expressed in terms of unionized ammon ia were

significantly higher at the warmer temperature, by factors ranging from 3.5 to 6.2.

Knoph (1992) conducted acute toxicity tests at temperatures ranging from 2 to 17°C

using Atlantic salmon parr, one series of tests at pH≈6.0 and the other at pH≈6. 4. In both series

of tests, LC 50 

## s expressed in terms of unionized ammonia increased substantially with

temperature.

Even with these additional data, the shape of the temperature relationship is not

completely resolved, especially for chronic toxi city. Nevertheless, the acute data for fishes

overwhelmingly indicate that ammonia toxicity, expressed in terms of unionized ammonia,

decreases with increasing temperature.

## 203

Figure M

. 1. The E ffect of T emperature on A mmonia T oxicity in T erms of U nionized A mmonia (DeGraeve et al. 1987)

. Symbols denote LC 50 

## s or LC 20 

## s and 95% confidence limits and lines denote linear regressions of log LC versus temperature.

## 204

Most importantly, the data of DeGraeve et al. (1987) show (Figure M

. 1) that (a) a linear

relationship of log unionized ammonia LC 50 

## versus temperature applies within the reported

uncertainty in the LC 50 

## s over the range of 6 to 30°C and (b) temperature effe cts on long

- term

mortality are similar to those on acute mortality. For invertebrates, acute toxicity data suggest

that ammonia toxicity, when expressed in terms of unionized ammonia, does not decrease, and

possibly even increases, with increasing tempera ture. Quantifying and adjusting data for this

relationship is not necessary because even at warm temperatures invertebrates are generally more

resistant to acute ammonia toxicity than fishes and thus their precise sensitivities are of limited

importance t o the criterion. At low temperatures, they are even more resistant relative to fishes

and thus their precise sensitivity is even less important to the criterion.

Based on this information, the two issues raised above were resolved as follows:

1. TCAPs will not be used in the ammonia criterion. This does not mean that the notion of

high temperature exacerbating ammonia toxicity is wrong; rather, it reflects the fact that

such an effect is not evident in the available data, which cover a wide temperature rang e.

2. A CMC, if it were expressed as unionized ammonia (rather than total ammonia, used in

this document) would continue to be lower at lower temperatures, consistent with the

observed temperature dependence of ammonia toxicity to the most sensitive species, i.e.,

fishes. Although it is possible that the temperature relationship differs among fish species

and that using the same relationship for all fish species introduces some uncertainty,

specifying a relationship for each fish species is not possible with c urrent data and would

also introduce considerable uncertainty.

Therefore, for a criterion expressed in terms of unionized ammonia, available data

support the continued use of a generic temperature relationship similar to that in the 1984/1985

ammonia criteria document, but without TCAPs.

## 205

Figure M

. 2. The E ffect of T emperature on A cute A mmonia T oxicity in T erms of T otal A mmonia Nitrogen

. Symbols denote LC 50 

## s, solid lines denote regressions for individual datasets, and dotted lines denote pooled regressions over all

datasets.

## 206

Figure M

. 3. The E ffect of T emperature on pH

- A djusted A cute A mmonia T oxicity in T erms of T otal A mmonia Nitrogen

. LC 50 

## s are adjusted to the mean pH of the dataset based on the pooled relationship of acute toxicity to pH. Symbols denote LC 50 

## s, solid

lines denote regressions for individual datasets, and dotted lines denote pooled regression over all datasets

## 207

This raised a new issue, ho wever, because the criterion expressed in terms of total

ammonia is nearly constant over all tested temperatures, and the small effect of temperature on

the total ammonia criterion in the 1984/1985 criteria document is largely an artifact of

conducting reg ression analyses in terms of unionized ammonia and is not indicative of any

established, significant trend. It was thought that the expression and implementation of the

ammonia criterion might have been simplified if temperature were dropped as a modifyin g

factor, which might have been possible if ammonia toxicity is expressed in terms of total

ammonia. Furthermore, permit limits and compliance are usually expressed in terms of total

ammonia nitrogen, and so expressing the criterion in terms of total ammo nia nitrogen would

simplify its implementation by eliminating conversions to and from unionized ammonia.

Because of such benefits and because there are no compelling scientific or practical reasons for

expressing the criterion in terms of unionized ammoni a, the freshwater toxicity data concerning

temperature dependence were reanalyzed in terms of total ammonia nitrogen.

The data analyzed are from the studies included in the 1984/1985 ammonia criteria

document and the studies of DeGraeve et al. (1987), Nim mo et al. (1989), and Knoph (1992).

All analyses were conducted in terms of total ammonia nitrogen, either as reported by the authors

or as converted by us from reported values for unionized ammonia, pH, and temperature using

the speciation relationship o f Emerson et al. (1975). The data are presented in Figure M

. 2 and

show considerable diversity, with some datasets showing decreasing toxicity with increasing

temperature, some showing increasing toxicity, and some showing virtually no change. There

are e ven differences among studies using the same test species. However, in no case is the effect

of temperature particularly large, being no more than a factor of 1.5 over the range of any

dataset, except for the Johnny darter data of Nimmo et al. (1989). In some studies, test pH was

correlated with test temperature. To reduce the confounding effect of pH, the total ammonia

LC 50 

## was adjusted to the mean pH of the data for the study using the pH relationship discussed in

the next section of this appendix . Th ese adjusted data are shown in Figure M

. 3 and also show

neither large effects nor any clear consistency among or within species or studies.

## 208

For each dataset containing at least three data points, a linear regression of log LC 50



## versus temperature was conducted (Draper and Smith 1981) and the resulting regression lines are

plotted as solid lines in Figures M

. 2 and M

. 3. These regressions are significant at the 0.05 level

for only one dataset (the unadjusted fathead minnow data of Thurston et al. 1983); for this

dataset, however, the regression is not significant when the data are adjusted for the fact that pH

value s were lower in the low

- temperature tests than in the high

- temperature tests. Slopes from

regression analyses of datasets in Figure M

. 3 range from

- 0.015 to 0.013, compared to a range

from 0.015 to 0.054 when expressed in terms of unionized ammonia (Erick son 1985). This

narrower range of slopes in terms of total ammonia nitrogen also argues for use of total

ammonia, rather than unionized ammonia, because there is less uncertainty associated with the

generic relationship. For datasets with just two points , Figures M

. 2 and M

. 3 also show the

slopes for comparative purposes. Based on the typical uncertainty of LC 50 

## s, these slopes also

would not be expected to be significant, except perhaps for the Johnny darter data of Nimmo et

al. (1989).

A multiple least

- squares linear regression (Draper and Smith 1981) using all datasets

(with a common slope for all datasets and separate intercept for each dataset) was conducted,

both with and without pH adjustment. The results of these pooled analyses are plotted as dot ted

lines in Figures M

. 2 and M

. 3 to show that the residual errors for the common regression line

compared to the individual regression lines are not large relative to the typical uncertainty of

LC 50 

## s. To better show the overall fit of the common regressio n line, the data are also plotted

together in Figure M

. 4 by dividing each point by the regression estimate of the LC 50 

## at 20°C for

its dataset. This normalization is done strictly for data display purposes because it allows all of

the datasets to be overl aid without changing their temperature dependence, so that the overall

scatter around the common regression line can be better examined. The data show no obvious

trend, with the best

- fit slope explaining only 1% of the sum of squares around the means for the

pH

- adjusted data and 0% for the unadjusted data. The one available chronic dataset (DeGraeve

et al. 1987) also shows no significant temperature effect when expressed in terms of total

## 209

ammonia nitrogen (Figure M

. 5) and adjusted for pH differences among the tests. (These tests

and the calculation of the LC20s are discussed in detail later.)

Based on the small magnitude and the variability of the effect of temperature on total

ammonia acute and chronic toxicity values for fish, the 1998 Update did not i nclude temperature

as a modifying factor for a total ammonia criterion. For invertebrates, it should be noted that the

1998 Update’s assumption that temperature had no effect on the toxicity of total ammonia differs

from the 1984/1985 criteria document’s assumption that temperature has no effect on the toxicity

of unionized ammonia. This inconsistency is resolved during the 1999 re

- examination of data, to

be discussed shortly, by incorporating a relationship between temperature and total ammonia

toxicity to invertebrates. That relationship, however, does not affect the (1999 update) CMC

because invertebrates are not among the acutely sensitive taxa.

## 210

Figure M

. 4. The E ffect of T emperature on N ormalized A cute A mmonia T oxicity in T erms of T otal A mmo nia Nitrogen

. Data were normalized by dividing measured LC 50 

## s by regression estimates of LC 50 

## s at 20°C for individual datasets for Figure M

. 2

(top plot) and Figure M

. 3 (bottom plot).

## 211

Figure M

. 5. The E ffect of T emperature on C hronic A mmonia L ethality to F athead M innows in T erms of T otal A mmonia

Nitrogen (DeGraeve et al. 1987).

Symbols denote LC20s and 95% confidence limits and lines denote linear regressions of log LC versus temperature. Figure on left is

for estimated LC 50 

## s at test pH and figure on right is for LC 50 

## s adjusted to pH=7.5 based on pooled relationship of chronic toxicity to

pH

.

## 212

The amount of uncertainty in this approach to the CMC can be demonstrated to be small

by considering how the criterion would differ if total ammonia toxicity was adjusted based on

the slopes in various datasets. Because the bulk of the toxicity data used in the derivation of the

criterion is within a few degrees of 20°C, the te mperature relationship used has very little effect

on the criterion near this temperature, but rather has the greatest effect on the criterion at much

higher or lower temperatures. If the average slope for the pH

- adjusted acute data from Figure

M

. 4 is use d, the total ammonia CMC at 5°C would be only about 6% higher than at 20°C. The

smallest and largest slopes from the acute regressions for individual species in Figure M. 3 would

produce a range from 40% lower to 68% higher at 5°C than at 20°C, but this gr eatly overstates

the uncertainty because effects on a CMC derived from many datasets should not be near these

extremes.

1999 Re

- examination of Temperature Dependence

– Acute Toxicity

The previous section, reproduced with relatively few changes from the 1 998 Update,

included an analysis of available data on the temperature dependence of acute ammonia toxicity

to fish. These data (in Figures M. 2, M. 3, and M. 4) consisted of 20 different data sets drawn from

11 different studies and included nine different s pecies, four of these species being in more than

one study. Data from Arthur et al. (1987) were not used in the 1998 analysis because those

authors reported concerns about factors confounding temperature in their data set. Linear

regression analysis of l og LC 50 

## (total ammonia basis) versus temperature was conducted on each

data set, both with and without correcting for pH as a confounding factor. No consistent trend

with temperature was observed and only one data set showed a slope different than zero at the

0.05 level of statistical significance. Therefore, a pooled linear regression analysis was

conducted across all data to derive an average slope, which was very close to zero and also not

statistically significant. On the basis of this analysis, the 1998 Update did not include any

temperature dependence for criteria to protect fish from acute ammonia toxicity.

In response to public comment (U.S. EPA, 1999), the 1998 analysis was re

- examined.

This re

- examination indicated that it is appropriate to h andle the temperature dependencies of

fish and invertebrates separately. For invertebrates, the inclusion of the Arthur et al. (1987) data

in the regression analysis yields a change in the temperature dependency that is ultimately

reflected in the differe nce between the 1999 CCC and the 1998 CCC.

## 213

In the 1998 Update, EPA did not use the Arthur et al. (1987) data because of those

authors concerns that other variable factors in their tests, conducted during different seasons,

might have had a potential to confound their results. In re

- examining their data in response to

comments, however, EPA found that most of the fish data from Arthur et al. showed behavior

similar to that from numerous other investigators: that is, little relationship with temperature

w hen expressed as total ammonia. Consequently, it was concluded that the other variable factors

were unlikely to be confounding the results.

For fish, although the temperature dependency is unchanged from 1998, additional

documentation is provided here, p rimarily because the apparent difference between fish acute

and chronic temperature dependencies is now used in the projection of the invertebrate chronic

temperature dependency.

First presented here will be more details on the regression analyses of th e individual data

sets conducted for the Update, plus similar analyses of the data of Arthur et al. A linear

regression was conducted on ea ch data set using the equation:

log (LC50 T 

## ) = log(LC50 20 

## ) + S (T

- 2 0)

where LC50 T 

## is the total ammonia LC50 at temperature T, S is the slope of log LC50 versus

temperature, and LC50 20 

## is the estimated total ammonia LC50 at 20°C. For completeness, this

effort included data sets with just two points, although the regression analysis the n provides a

perfect fit and has no residual error, so that confidence limits, significance levels, etc. cannot be

evaluated using normal methods. In such cases, the mean squared error (MSE) of data around

the regression was assumed to be equal to the wei ghted mean residual MSE for the larger data

sets, so that approximate significance levels could be determined.

Fish acute data:

Table M

. 1 presents the results of the regression analysis for each data set, with data

adjusted to pH=8 based on the average pH relationship used in the 1998 and 1999 Updates.

Plots of these relationships (except for Arthur et al. 1987) are in Figure M

. 3 in the p revious

section.

## 214

Of the 24 entries in Table M

. 1, nearly half (11) have very small slopes of between

- 0.006

and +0.006, a range which corresponds to a factor of 1.3 change or less in LC 50 

## for a 20°C

temperature change and is less than normal data variabili ty. Of these 11, five have positive

slopes and six have negative slopes. Of the 13 entries with steeper slopes, five have positive

slopes and eight have negative slopes. Among the data sets used in the Update, only two of the

regressions are statistical ly significant at the 0.05 level, one with a negative slope and one with a

positive slope, although two other sets (for fathead minnows from DeGraeve et al. 1987 and

Reinbold and Pescitelli 1982 b ) are close to being significant. (The level of significance for the

Johnny darter data set differs from what was reported in the previous section because it consists

of two different sets which were analyzed separately in the 1998 analysis, but combined here

because they were not significantly different.) Of the five data sets from Arthur et al. (1987),

only one is significant at the 0.05 level. For species with more than one entry, slopes vary

considerably. This general lack of statistical significance and consistency precludes any reliable

assessments based on these individual analyses.

The 1998 Update therefore conducted a pooled regression analysis to determine whether

the combined acute toxicity data sets indicated any significant average trends with temperature.

Table M

. 2 summarizes the mean trends dete rmined in various pooled analyses. The first entry is

the pooled analysis conducted for the 1998 Update, which included all the data in Table M

. 1

above except the fish data of Arthur et al. (1987). The slope from this pooled analysis was very

small (

- 0.0 023), and was not statistically significant despite the large number of data. The

second entry adds the fish data of Arthur et al.; it does result in a statistically significant trend.

The mean slope (

- 0.0058) is still small, but does amount to a 23% dec rease in LC 50 

## per 20°C

increase in temperature. However, this slope is heavily influenced by two points with high

residual (>3σ) deviations. One of these points is a test at 3.4°C by Arthur et al. (1987) with

fathead minnows, which showed much greater ef fects of low temperature than other studies with

the same species. The other point is for a test at 22.6°C by Arthur et al. (1987) with walleye,

which showed very high sensitivity and was part of a set of three tests which used fish from

different sources , potentially confounding the temperature effects. Without these two data, the

regression has an even lower slope and is not significant at the 0.05 level (third entry in Table

M. 2). Overall, these analyses of the fish acute data suggest a weak overall t rend of higher LC 50 

## s

## 215

at low temperatures, with a logLC 50 

## versus temperature slope in the

- 0.002 to

- 0.006 range, but

of questionable statistical significance.

Table M.1. Results of R egression A nalysis of logLC 50 

## (mg/L total ammonia nitrogen )

V ersus T emperature (°C) for I ndividual D ata S ets on the T emperature D ependence of

A cute A mmonia T oxicity.

Table M.1

Reference/

Species Slope

(95% CL ) logLC50 20 (95% CL ) Residual SD

(r 2 ) F REGR

(α)

Thurston et al. (1983)

Fathead Minnow - 0.0014

(

- 0.013,+0.013) 1.641

(1.582,1.700) 0.112

(<1%) 0.06

(0.81)

Thurston and Russo (1983)

Rainbow Trout +0.0059

(

- 0.017,+0.029) 1.350

(1.204,1.495) 0.121

(2%) 0.30

(0.59)

Cary (1976)

Channel Catfish +0.0028

(

- 0.008,+0.013) 1.676

(1.593,1.758) 0.093

(2%) 0.32

(0.58)

Colt and Tchobanoglous (1976)

Channel Catfish +0.0004

(

- 0.037,+0.038) 1.604

(1.350,1.858) 0.016

(2%) 0.02

(0.91)

Ministry of Technology (1967)

Rainbow Trout +0.0008

(

- 0.018,+0.019) 1.231

(1.010,1.452) 0.051

(1%) 0.03

(0.88)

Roseboom and Richey (1977)

Bluegill Sunfish +0.024

(

- 0.025,+0.073) 1.089

(0.803,1.375) - 0.95

(0.33)

Roseboom and Richey (1977)

Channel Catfish +0.020

(

- 0.029,+0.069) 1.482

(1.196,1.768) - 0.68

(0.41)

Roseboom and Richey (1977)

Largemouth Bass - 0.0029

(

- 0.040,+0.034) 1.237

(0.972,1.502) - 0.02

(0.88)

Reinbold and Pescitelli (1982 b )

Rainbow Trout - 0.0088

(

- 0.028,+0.010) 1.396

(1.159,1.632) 0.088

(29%) 1.63

(0.27)

Reinbold and Pescitelli (1982 b )

Bluegill Sunfish - 0.0004

(

- 0.027,+0.026) 1.370

(1.059,1.681) 0.128

(0%) 0.00

(0.96)

Reinbold and Pescitelli (1982 b )

Fathead Minnow - 0.0153

(

- 0.031,+0.009) 1.429

(1.243,1.615) 0.076

(89%) 16.6

(0.06)

Hazel et al. (1971)

Striped Bass - 0.0163

(

- 0.057,+0.025) 1.274

(1.105,1.443) 0.076

(60%) 2.93

(0.23)

Hazel et al. (1971)

Three

- Spined Stickleback - 0.0106

(

- 0.053,+0.032) 1.390

(1.214,1.567) 0.081

(36%) 1.14

(0.40)

DeGraeve et al. (1987)

Fathead Minnow - 0.0052

(

- 0.012,+0.002) 1.617

(1.563,1.670) 0.061

(36%) 3.33

(0.12)

DeGraeve et al. (1987)

Channel Catfish - 0.0088

(

- 0.016,

- 0.002) 1.648

(1.595,1.701) 0.061

(62%) 9.76

(0.02)

Knopf (1992)

Atlantic Salmon - 0.0035

(

- 0.027,+0.020) 1.715

(1.406,2.025) 0.097

(7%) 0.22

(0.067)

Knopf (1992)

Atlantic Salmon +0.0163

(

- 0.075,+0.108) 1.636

(0.405,2.866) 0.054

(84%) 5.18

(0.26)

Nimmo et al. (1989)

Johnny Darter +0.021

(+0.000,+0.043) 1.463

(1.248,1.678) 0.072

(90%) 18.1

(0.05)

Nimmo et al. (1989)

Fathead Minnow +0.0070

(

- 0.014,+0.028) 1.568

(1.353,1.782) - 0.42

(0.52)

Arthur et al. (1987)

Fathead Minnow - 0.032

(

- 0.059,

- 0.004) 1.762

(1.493,2.030) 0.105

(92%) 24.8

(0.04)

## 216

Table M.1

Reference/

Species Slope

(95% CL ) logLC50 20 (95% CL ) Residual SD

(r 2 ) F REGR

(α)

Arthur et al. (1987)

Rainbow Trout - 0.0100

(

- 0.053,+0.033) 1.348

(0.937,1.758) 0.158

(16%) 0.56

(0.51)

Arthur et al. (1987)

Channel Catfish - 0.0058

(

- 0.038,+0.027) 1.558

(1.230,1.886) 0.030

(84%) 5.15

(0.26)

Arthur et al. (1987)

White Sucker +0.0007

(

- 0.23,+0.25) 1.902

(1.657,2.147) 0.048

(1%) 0.01

(0.92)

Arthur et al. (1987)

Walleye - 0.038

(

- 0.327,+0.250) 1.216

(

- 1.911,4.343) 0.306

(74%) 2.84

(0.34)



## Table M

. 2. Results of R egression A nalysis of log LC 50 

## (mg/L total ammonia nitrogen )

V ersus T emperature (°C) for P ooled D ata S ets on the T emperature D ependence of A cute

A mmonia T oxicity to F ish.

Data Sets Pooled Slope

(95% CL ) Residual SD

(r 2 ) F REGR (α)

All Data excluding Arthur et al.

- 0.0023

(

- 0.0057,+0.0011) 0.105

(2%) 1.79

(0.18)

All Data including Arthur et al.

- 0.0058

(

- 0.0094,

- 0.0022) 0.122

(8%) 10.3

(<0.01)

All Data including Arthur et al. except "Outliers"

- 0.0030

(

- 0.0063,+0.0002)) 0.105

(3%) 3.52

(0.06)

Fathead Minnow excluding Arthur et al

- 0.0063

(

- 0.0122,

- 0.0005) 0.106

(11%) 4.76

(0.04)

Fathead Minnow including Arthur et al.

- 0.0105

(

- 0.0169,

- 0.0049) 0.120

(25%) 13.4

(<0.01)

Fathead Minnow including Arthur et al. exc l "Outlier"

- 0.0073

(

- 0.0129,

- 0.0017) 0.106

(15%) 6.85

(0.01)

Rainbow Trout excluding Arthur et al.

- 0.0013

(0.0122,+0.0096) 0.109

(<1%) 0.06

(0.80)

Rainbow Trout including Arthur et al.

- 0.0034

(

- 0.0133,+0.0064) 0.115

(2%) 0.51

(0.48)

Channel Catfish excluding Arthur et al.

- 0.0030

(

- 0.0091,+0.0031) 0.088

(4%) 1.05

(0.32)

Channel Catfish including Arthur et al.

- 0.0034

(

- 0.088,+0.021) 0.085

(6%) 1.64

0.21)

Bluegill Sunfish +0.0006

(

- 0.0172,+0.0184) 0.120

(<1%) 0.01

(0.92)

## 217

It is also useful to consider separately the overall trends for different fish species. Table

M

. 1 includes multiple studies with fathead minnows, rainbow trout, channel catfish, and bluegill

sunfish. Table M

. 2 includes the results of pooled analyses for each of these species, both with

and without data from Arthur et al. (1987). For rainbow trout , bluegill, and channel catfish, the

regressions were not statistically significant. The bluegill data indicated virtually no temperature

effect, whereas weak trends similar to the pooled analyses over all data sets were suggested in

the channel catfish d ata (slope =

- 0.0030 without and

- 0.0034 with Arthur et al. data) and

rainbow trout data (slope =

- 0.0014 without and

- 0.0034 with Arthur et al. data). For fathead

minnow, the pooled analyses were statistically significant and stronger, with slopes rangin g from

- 0.0063 to

- 0.0105 depending on the treatment of data from Arthur et al. Such slopes for fathead

minnow would result in moderate effects over a broad temperature range: a 20°C decrease in

temperature would result in a 33% to 62% increase in LC 50



## . However, this species is not

sensitive enough that this would affect the acute criterion values. For the species used in the

acute criterion calculations, no temperature correction for acute toxicity is appropriate due to the

lack of any significant tren d over all data sets.

Invertebrate acute data:

Unlike fish, available acute toxicity data for invertebrates indicates that their acute

sensitivity to ammonia decreases substantially with decreasing temperature. The 1998 Update

noted this temperature dep endence, but did not present any analysis of it because tested

invertebrates were sufficiently tolerant to acute ammonia exposures that this dependence would

not affect the acute ammonia criterion. The 1998 Update also noted that this temperature

dependen ce should be a consideration in setting low temperature chronic criterion, but did not

provide any specific analysis regarding this. This section will provide an analysis of available

information on the temperature

- dependence of invertebrate acute ammonia toxicity, to be used

later for estimating the temperature

- dependence of chronic ammonia toxicity.

Arthur et al. (1987) provide the only available data on the temperature dependence of

acute ammonia toxicity to invertebrates. As noted earlier, these toxi city tests did not specifically

test temperature effects, but rather were seasonal tests in which various chemical characteristics

of the tests water varied as well as temperature. Test organisms were whatever were available in

outdoor experimental stream s at the time of the test, so the size, life stage, and condition of the

## 218

organisms also varied. The authors of this study expressed some doubt as to how much of the

effects they observed were actually due to temperature. However, for invertebrates, they did

observe strong correlations of total ammonia toxicity with temperature. Confounding factors

might contribute somewhat to this correlation, but temperature is still likely the primary

underlying cause. If other factors were largely responsible for the apparent effects of

temperature, it would be expected that strong correlations with temperature would also be

evident in their fish data. However, as discussed above, the fish data usually showed much

weaker effects of temperature, similar to other studi es with fewer confounding factors.

These invertebrate acute data were analyzed using the same regression model and

techniques as discussed above for fish. The study of Arthur et al. (1987) included data sets for

nine invertebrate species, but two of thes e sets were not included in the analysis because they

consisted of two tests at temperatures only 3°C apart. For the other species, the number of tests

ranged from 2 to 6, with temperature ranges of from 9°C to 21°C. Table M

. 1 summarizes the

regression r esults for the data sets of each species and for pooled analyses conducted on (a) all

seven species, and (b) three species that had more than two tests and a temperature range of at

least 15°C. All data were corrected to pH 8 based on the average acute pH relationship

(described later). All species show substantially greater tolerance to ammonia at lower

temperatures, and in most cases the significance level of the regression is better than 0.05. (As

for the analysis of the fish data, when there were jus t two tests for a species, the significance

level for the individual analysis is based on the MSE from the pooled analysis.) The slope of log

LC 50 

## versus temperature does not vary widely, ranging from

- 0.028 to

- 0.046 and being

- 0.036

for both pooled anal yses. Figure M. 6 provides plots of this data and the regression lines

comparable to those for fish previously shown in Figures M

. 3 and M

. 4.

Again, because invertebrates are not among the species acutely sensitive to ammonia (in

the 1999 update) , the inve rtebrate acute temperature slope does not affect the formulation of the

acute criterion. It will be used subsequently, however, in formulating the invertebrate chronic

temperature slope, which ultimately will affect the formulation of the chronic criterio n.

## 219

1999 Re

- examination of Temperature Dependence

– Chronic Toxicity

Fish chronic data

As in the 1998 Update, the only available data on the temperature dependence of chronic

ammonia toxicity are from the study by DeGraeve et al. (1987) on survival of juvenile fathead

minnows during 30

- day exposures to ammonia at temperatures ranging from 6° C to 30°C. In

contrast to acute toxicity, which for fathead minnows showed sensitivity to be slightly reduced at

low temperatures, this data on chronic toxicity suggested greater sensitivity at low temperatures.

However, this trend was small, at least on ce the confounding effect of pH was corrected for, and

not statistically significant. Based on this analysis, the 1998 Update treated effect concentrations

for chronic ammonia toxicity to fish as it did for acute toxicity: as being invariant with

tempera ture. However, the 1998 Update also noted that, if seasonal variations in temperature

cause a shift in what endpoints the criterion should be based on, the chronic criterion could have

a seasonal temperature dependence even if effect concentrations for sp ecific chronic endpoints

do not vary with temperature (This is discussed in the 1999 AQWC document under the section

named S easonality of Chronic Toxicity Endpoints )

.

This section will provide more details regarding the analysis of the chronic toxicity da ta

from DeGraeve et al. (1987), and a comparison of its temperature dependence to that of acute

toxicity in the same study. Figure M

. 5 showed the temperature dependence of acute and chronic

effect concentrations from this study.

An important issue in thi s analysis is the confounding effect of pH on the apparent effect

of temperature, because pH increased with decreasing temperature in these chronic exposures.

To examine what the effect of temperature is, the effect concentrations should be adjusted to a

common pH using an equation that accounts for the effect of pH. A critical question then is what

pH equation to use, because no study exists for the effect of pH on this particular chronic

endpoint (juvenile 30

- day survival), or on the interaction of pH a nd temperature effects. The

1998 Update used the pH relationship derived for the chronic criterion. Of the pH relationships

available, that one is probably most appropriate, but entails some uncertainty. To evaluate how

conclusions about temperature eff ects will vary if the true pH relationship is different, this

analysis will also use the pH relationship for acute toxicity to fathead minnows from Thurston et

al. (198 3 ). This relationship likely represents an extreme possibility; i.e., it assumes that c hronic

## 220

toxicity pH relationships are the same acute ones, contrary to what is indicated by available

chronic studies.

## 221

Figure M

. 6. Temperature Dependence of Acute Ammonia Toxicity to Invertebrate

Organisms from Arthur et al. (1987)

.

## 222

Using the pooled chronic pH relationship (presented later in this document), slope=0.010,

significance=0.13, and r 2 

## =0.76. Using the fathead minnow acute pH relationship, slope=0.0053,

significance=0.32, and r 2 

## =0.45. In neither case is the regression statistically significant at the 5

percent level, due to the amount and variability of the data. Nevertheless, it should be noted that,

in both cases, the chronic data show an upward trend with temperature, in con trast to that

observed for acute toxicity. Even under the extreme assumption that these data have a pH

relationship similar to acute toxicity, the slope is 0.005, and is twice this under the assumption

that these data follow the chronic pH relationship. Thus, even if fathead minnows show

increased acute tolerance to ammonia at low temperatures, a similar assumption for chronic

toxicity is contraindicated.

The difference between acute and chronic temperature relationships can be better

assessed by looking at acute

- chronic ratios. Figure M

. 7 shows the temperature dependence of

the ratio of the acute LC 50 

## to the chronic LC 20 

## . The chronic LC 20 

## s used for the ACRs were

normalized via the above two alternative pH relationships, while the acute LC 50 

## s were

norma lized only using the acute pH relationship. The results show that for either pH

normalization alternative, the ACRs are substantially higher at lower temperatures than at higher

temperatures. If the chronic data are pH

- normalized using the chronic pH rel ationship, the

regression is significant at the 0.05 level, with a slope of

- 0.0155. If normalized using the acute

pH relationship, the slope is less (

- 0.0110), but even with this extreme assumption, there is only a

13 percent probability that the regress ion slope arose by chance.

It is not surprising that acute

- chronic ratios are higher at low temperature. Temperature

can affect toxicity in a variety of ways, one of which is simply to slow down responses. This is

evident in some reports on the effect o f temperature on ammonia toxicity. For example, for the

rainbow trout data from Ministry of Technology (1967), there was little effect of temperature on

total ammonia LC 50 

## s at 96 hours, but at shorter durations LC 50 

## s increased with decreasing

temperature. The overall impact on the temperature dependence of LC 50 

## s and ACRs will depend

on the duration of the acute toxicity test and on the speed of action of acute ammonia toxicity in

the species of concern. However, temperature is likely to affect ammonia to xicity in multiple

ways, some of which would alter acute and chronic toxicity similarly. Nonetheless, to some

degree the ratio of effect concentrations at different durations is expected to increase at lower

## 223

temperatures. This expectation, as well as the empirical evidence, argues against the direct

application of acute temperature relationships to chronic toxicity.

Figure M

. 7. Temperature

- D ependence of A mmonia ACRs for F athead M innows.

(The choice of reference condition, pH =8 here versus pH=7.5 in Figu re M

. 5, has no effect on

slope or significance.)

Invertebrate chronic projections

No data are available on the effect of temperature on chronic ammonia toxicity to

invertebrates. Because invertebrates are much more acutely tolerant at low temperatures than at

high temperatures, it is likely that their chronic toxicity would also show some temperature

dependence. However, as discussed above, there is reason to expect acute and chronic toxicity to

vary somewhat differ ently with temperature, with acute

- chronic ratios increasing at low

temperature, especially for organisms for which acute ammonia toxicity is not especially fast,

which is the case for invertebrates (Thurston et al. 1984 b ). The observed trend in the fathe ad

minnow ACRs provides support for this expectation.

The critical question then becomes, how much of the acute temperature slope for

invertebrates should be assumed to apply to chronic toxicity? If this slope is predominantly due

## 224

to temperature

- induced delays of acute toxicity, chronic toxicity might have very little slope. If

this slope is not at all due to such delays, then all the slope should be applied to chronic toxicity.

One option for an objective mathematical prediction of the invertebrate chr onic slope is

to assume that the difference between acute and chronic slopes will be the same for fish and

invertebrates, potentially implying that the effect of temperature on the kinetics of toxicity is

roughly the same for fish and invertebrates. In th is case the invertebrate chronic slope would be

the difference between

- 0.036, the average invertebrate acute slope, and

- 0.016, the observed

slope for fish acute

- chronic ratios. This would yield an invertebrate chronic slope of

- 0.020.

This correction s till applies most of the acute slope to chronic toxicity, but recognizes that the

chronic slope should probably be less steep.

It is recognized that few data are available to define the Figure M

. 7 fish ACR slope, and

that the assumption the invertebrate A CR slope would equal the fish ACR slope is quite

uncertain despite having some theoretical underpinning in the kinetics of toxicity. Consequently

a second option is to equate the invertebrate chronic slope to the invertebrate acute slope (

- 0.036)

minus on e

- half the fish ACR slope (

- 0.016/2). This splits the difference between no correction

and full correction for the fish ACR slope, resulting in an invertebrate chronic slope of

- 0.028.

A third, related option is suggested from the appearance of data in t he last two plots in

Figure M

. 6, plots of “All Species” and “Physa, Crangonyx, Musculium”. These plots suggest a

steeper invertebrate acute slope at higher temperatures than at very low temperature. At greater

than 10°C, these data also comfortably fit a slope of

- 0.044. If such a slope were used to fit those

data, however, a concentration plateau would need to be imposed between 5 and 10°C to avoid

over

- estimating the acute effect concentrations measured near 5°C. If the invertebrate chronic

slope is o btained by subtracting the full value of the fish ACR slope (

- 0.016), this would yield

the same invertebrate chronic slope,

- 0.028, as the option in the previous paragraph. In this case,

however, concentrations would be capped between 5 and 10°C in order to reflect the implied

attenuation of slope at low temperature relative to higher temperatures.

EPA selected this third option, a compromise between the first two options, for defining

the invertebrate chronic temperature slope in formulating the CCC, dis cussed later. This

provides a good fit to the available information.

## 225

Appendix N. Site

- Specific Criteria for Ammonia

.

Recalculation Procedure for Site

- specific Criteria Derivation

The water quality standards (WQS) regulation at 40 CFR § 131.11(b)( 1)(ii) provide s

states with the opportunity to adopt water quality criteria that are “...modified to reflect site

specific conditions.” As with any criteria, site

- specific criteria must be based on a sound

scientific rationale in order to protect the design ated use and are subject to review and approval

or disapproval by EPA.

The recalculation procedure for site

- specific criteria derivation is intended to allow site

specific criteria that differ from national criteria recommendations (i.e., concentrations th at are

higher or lower than national recommendations) where there are demonstrated differences in

sensitivity between the aquatic species that occur at the site and those that were used to derive

the national criteria recommendations. The national dataset may contain aquatic species that are

sensitive to a particular pollutant, but these or comparably sensitive species might not occur at

the site (e.g., freshwater mussels are included in the national ammonia dataset but may not be

present at a particular s ite). On the other hand, a species that is critical at the site might be

sensitive to the pollutant and require site

- specific criteria that are lower than the national

recommended criteria.

In the case of ammonia, where a state demonstrate s that mussels are not present on a site

specific basis, the recalculation procedure may be used to remove the mussel species from the

national criteria dataset to better represent the species present at the site. For example, many of

the commonly occurring freshwater b ivalves (e.g., pea clam) are more closely related to the non

unionid fingernail clam Musculium (which is the fourth most sensitive genus in the national

dataset for the chronic criterion) than to the unionid mussels Lampsilis and Villosa ( which are the

two most sensitive genera in the national dataset for the chronic criterion) . At sites where all

bivalves present are more closely related to Musculium than to Lampsilis and Villosa (i.e., where

unionid mussels are not present at the site), the recalculat ion procedure may be used to remove

Lampsilis and Villosa from the dataset because they would not be representative of the species

present at the site. With removal of Lampsilis and Villosa from the national dataset , the

recalculation procedure could resu lt in criteria (and associated water quality

- based effluent limits

(WQBELs) based on such criteria) with higher concentrations than EPA’s recommendations but

## 226

that are still protective of the designated use

. The retention of Musculium in the dataset would

represent the other non

- unionid bivalves present at the site, so the non

- unionid bivalves would

still be protected if Lampsilis and Villosa were removed from the chronic dataset. However, at

sites where both unionid and non

- unionid bivalves are present, a ll three bivalves in the national

chronic dataset (i.e., Lampsilis

, Villosa , and Musculium ) would be retained because they would

represent the species present at the site. The recalculation procedure describes how to compare

the taxonomy of species presen t at the site with the taxonomy of species in the national dataset.

The number of tested genera (N) in the criteria calculations must be updated where

genera such as Lampsilis and Villosa are removed from the datas et

. For example, if only the two

unionid mussels are removed from the dataset for the national chronic ammonia criterion, N

would be reduced from 16 genera in the national dataset to 14 genera in the site

- specific dataset

,

and this would affect the site

- specific criteria values

.

Freshwater snail s represent another sensitive freshwater species group for which acute

and chronic toxicity data exist and are used in criteria derivation. Because freshwater snails tend

to be more ubiquitous in the environment, however, the existing data for these anima ls are not

likely to be deleted from the datasets in a criteria recalculation.

As with any criteria, states choosing to utilize the recalculation procedure should ensure

that their site

- specific criteria “...provide for the attainment and maintenance of the water quality

standards of downstream waters.” 40 CFR § 131.10(b). In addition, states should consider how

they will demonstrate that mussels are not present at the site before selecting this approach. For

additional information on the recalculation pro cedure, see EPA’s Water Quality Standards

Handbook at http://www.epa.gov/wqshandbook

.

Acute Criterion Magnitude Recalculation for Ammonia

Unionid Mussels Present and Oncorhynchus species Absent

Where Oncorhyn chus sp ecies are absent, EPA does not lower its acute criteria for

ammonia below the 5 th 

## percentile in order to protect the commercially and recreationally

important adult rainbow trout , but instead, retains all tested species in the Order Salmoniformes

as tested surrogate species representing untested freshwater fish resident in the U.S. from another

Order

. The lowest GMAV for a freshwater fish ( vertebrate species ) is 51.93 mg TAN/L for

Prosopium (Table 3). Therefore, in this case, the CMC equals the lower of: a) 0.7249 times the

## 227

temperature adjusted lowest invertebrate GMAV (e.g., 17 mg TAN/L at pH 7.0 and 20o C) , or (b)

0.7249 times the lowest freshwater fish GMAV (e.g., 38 mg TAN/L at pH 7.0), according to the

following temperatur e relationship:

( ( ) 

## )

Thus, the CMC increases with decreasing temperature as a result of increased invertebrate

insensitivity until it reaches a plateau of 37.65 mg TAN/L at 10.2 °C and below (51.93 mg

TAN/L x 0.72 49), where the most sensitive taxa switches to the temperature invariant fish genus

Prosopium ( Table s 5b and N

. 1 ; see also Oncor h yn c hus absent line in Figure 5a ). Note: while the

mountain white fish ( Prosopium williamsoni ) is a species in the same family a s Oncorhynchus sp.

(i.e., Family: Salmonidae), it is also an appropriately sensitive surrogate species amongst all

freshwater fish in the Class Actinopterygii.

The CMC where Oncorhynchus sp. are absent extrapolated across both temperature and

pH is as fo llows:

( ) ( ( ) 

## )

When a threatened or endangered specie s occurs at a site and sufficient data indicate that it is

sensitive at 1

- hour average concentrations below the CMC, it is appropriate to consider deriving

a site

- specific criterion. It should be noted that the dataset used to derive these new ammonia

criteria included some threatened or endangered species, none of which were the most sensitive

of the species tested. E xtrapolated values across a range of temperatures and pH values are

presented in Table N

. 1

.

Unionid Mussels Absent and Oncorhynchus spp. Present

If a state can demonstrate that unionid mussels are not present at a site, a site

- specific

criteria can be cal culated for waters with mussels absent. It is important to recognize that for

site

- specific criteria derived where unionid mussels are absent, the commercially and

## 228

recreationally important adult rainbow trout ( Oncorhynchus mykiss ) is the most acutely sens itive

species. Thus, when Oncorhynchus spp. are present, the acute criterion cannot exceed 24 mg

TAN/L (the SMAV for adult rainbow trout 48.21 mg TAN/L divided by two

– see also Acute

Criterion Calculation section in this document).

At pH 7, the tempera ture relationship is expressed as follows:

( ( ( ) 

## ) )

Where 24.10 mg TAN/L is one half the SMAV of 48.21 mg TAN/L for adult rainbow trout, and

45.05 is 0.7249 (the CMC divided by the lowest GMAV in the complete acute dataset)

multiplied by 62.15 mg TAN/L, the GMAV of the temperature dependent pebblesnail

( Fluminicola sp.), the most sensitive non

- mussel invertebrate (Table N

. 2 ).

At temperatures 0

- 27.5°C, the CMC with mussels absent and Oncorhynchus sp p

.

present is 24.10 mg TAN /L, because adult rainbow trout remain the most sensitive species group

in this temperature range. At temperatures greater than 27.5°C, however, the GMAV for

Fluminicola sp ecies (62.15 mg TAN/L) becomes the most sensitive GMAV because

invertebrates are in creasingly more acutely

- sensitive to ammonia as temperature increases, and

thus, the CMC equals that of the mussels absent, Oncorhynchus sp. absent temperature

relationship (Figure N

. 1). Consistent with the approach followed with the unionid mussels

prese nt, Oncorhynchus sp ecies absent CMC calculation in the Acute Criterion Calculations

section of this document, the site

- specific criteria should 1) retains all tested species in the Order

Salmoniformes as tested surrogate species representing untested fresh water fish resident in the

U.S. from another Order; and 2) maintains the SSD relationship from the complete acute dataset

(i.e., CMC is equal to the lowest GMAV times 0.7249).

The CMC, where mussels are absent and Oncorhynchus spp. are present, extrapolat ed

across both temperature and pH is as follows (extrapolated values provided Table N

. 3 ):

( ( ) ( ( )

( ) 

### ) )

## 229

Unionid Mussels Absent and Oncorhynchus spp. Absent

If both unionid mussels and Oncorhynchus spp. are absent, the CMC calculated using the

Guidelines algorithm is 30.25 mg TAN/L at pH 7 and 20 ° C and is based on the four most

sensitive GMAVs in the following rank order: mountain whitefish, Lost River sucker,

pebblesnail, and golden shiner (see Table N.2 ). The ratio of the mussels absent and

Oncorhynchus spp. absent CMC to the most sensitive GMAV ( i.e., mountain whitefish;

Prosopium sp.) is 0.5825, or 30.25 mg TAN/L divided by 51.93 mg TAN/L. However, this

would result in a more protective criterion than when Oncorhynchus spp. are absent but mussels

are present (see Acute Criterion Calculations ). Because the unionid mussels absent and

Oncorhynchus spp. absent CMC cannot be more protective than the uni onid mussels present and

Oncorhynchus spp. absent CMC, the CMC to lowest GMAV ratio of 0.7249 from the complete

acute dataset is multiplied by 51.93 mg TAN/L for Prosopium sp., the lowest GMAV in the

unionid mussels absent dataset, resulting in a calculate d CMC of 37.65 mg TAN/L at pH 7 and

20 ° C . This is equivalent to the maximum plateau CMC when mussels are present and

Oncorhynchus spp. are absent at temperatures of 10.2°C and below (compare in Figures 5a and

N.1).

At pH 7, the temperature relationship i s expressed as follows:

( ( ( ) 

## ) )

At temperatures between 0

- 22.1°C the CMC with unionid mussels and Oncorhynchus

spp. absent is 37.65 mg TAN/L. At temperatures greater than 22.1°C, the temperature dependent

pebblesnail ( Fluminicola sp.) becomes the most sensitive GMAV, and the CMC decreases with

increasing temperature (Figure N.1).

The CMC, where both unionid mussels an d Oncorhynchus spp. are absent, extrapolated

across both temperature and pH is as follows (extrapolated values provided Table N

. 4 ):

( ) ( ( ( ) 

### ) )

## 230

A summary of the acute crite rion recalculations for all four mussel and Salmonid present and

absent combinations at pH 7 and 20°C is included in Table N

. 5.

Chronic Criterion Magnitude Recalculation for Ammonia

Unionid Mussels Absent, Early Life Stage (ELS) Protection Necessary

When unionid mussels are present, the CCC is the same regardless of whether early life

stages (ELS) of fish genera require protection. This is because unionid mussels represent the two

most sensitive genera in the chronic dataset, and at pH 7, the CCC at the invertebrate temperature

plateau of 7°C is 4.363 mg TAN/L, which is lower than the GMCV for Lepomis

, the most

sensitive fish genera, multiplied by the CCC to lowest GMCV ratio ( or 6.920 mg TAN/L x

0.8876 = 6.142 mg TAN/L

– see Chronic Criterion Calcula tions for additional details).

When unionid mussels are absent and fish ELS require protection, however, t he CCC is

6.508 mg TAN/L at pH 7 and 20 ° C (Table s N

. 6, N

. 7 ). The lowest GMCV is 6.920 mg TAN/L

for the temperature invariant vertebrate genus Lepomis , and the most sensitive invertebrate

GMCV is 7.547 mg TAN/L for Musculium (Table N

. 6 ). The ratio of the CCC to the most

sensitive GMCV ( Lepomis sp.) when unionid mussels are absent is 0.9405, or 6.508 mg TAN/L

divided by 6.920 mg TAN/L. At pH 7 and 20 ° C , the CCC when mussels are absent and ELS

protection is required is expressed as follows:

( ( ( ) 

## ) )

This function remains constant at a CCC equal to 6.508 mg TAN/L at 0

- 21.3 ° C because

the most sensitive GMCV is for the temperature invariant genera Lepomis (Figure N.2; Table

N

. 6 ). At temperatures greater than 21.3 ° C , the GMCV for the invertebrate Musculium (i.e.,

7.547 mg TAN/L) becomes the most sensitive, and the CCC decreases with increasing

temperature (Figure N.2 ).

When unionid mussels are absent and ELS protection is required, the thirty

- day average

concentration of ammonia nitrogen (in mg TAN/L) does not exceed, more than once every three

years on the average, the CCC calculated using the following equation:

## 231 ( ) ( ( ( ) 

### ) )



## Recalculated chronic criterion concentrations for the mussels absent, fish ELS protection

necessary scenario across a range of temperatures and pH values are provided i n Table N

. 8.

Unionid Mussels Absent, Early Life Stage (ELS) Protection Not Necessary

One approach for setting a chronic criterion for mussels absent and fish ELS absent is to

modify the criterion for mussels absent and fish ELS present . The four most sensitive genera for

the criterion to be modified are Lepomis (ELS), Musculium

, Fluminicola , and Pimephales (ELS),

which had yielded a criterion of 6.508 mg TAN/L at pH 7.0 and 20o C, or 0.9405 x the lowest

GMCV ( Lepomis ). Since the Lepom is GMCV, 6.920 mg TAN/L, is based on ELS sensitivity

,

consider that with ELS absent this value would increase to its juvenile and adult GMCV of 21.3

mg TAN/L (from U.S. EPA 1999, page 75 GMCVs, translated from pH 8 to pH 7). In this case,

Musculium , with GMCV 7.547 mg TAN/L, would now be the most sensitive genus in the

dataset, such that at pH 7 and 20 o C the criterion could be calculated as 0.9405 x 7.547 = 7.098

mg TAN/L. Because Musculium remains the most sensitive genus throughout the full range of

tem peratures, the criterion follows the invertebrate temperature relationship, increasing with

decreasing temperature until it reaches its maximum at the built

- in 7 oC plateau, which is 16.41

mg TAN/L at pH 7, fully protective of the lowest juvenile

- adult fish GMCV, that for Lepomis

,

21.3 mg TAN/L shown above.

Mussels absent ELS protection not required at pH 7

( ( ( ) ) 

## )

Overall

( ) ( ( ( ) ) 

## )

Recalcula ted c h ronic criterion concentrations for the mussels absent, fish ELS protection not

required scenario across a range of temperatures and pH values are provided in Table N

. 9. A

## 232

summary of the chronic criterion recalculations for all four mussel and fish EL S present and

absent combinations at pH 7 and 20°C is included in Table N

. 7.

## 233

T able N.1 . Temperature and pH

- Dependent Values of the CMC (Acute Criterion Magnitude )

– Unionid Mussels P resent,

Oncorhynchus A bsent

. Temperature ( ° C)

pH 0

- 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25 26 27 28 29 30



### 6.5 51 48 44 41 37 34 32 29 27 25 23 21 19 18 16 15 14 13 12 11 9.9



### 6.6 49 46 42 39 36 33 30 28 26 24 22 20 18 17 16 14 13 12 11 10 9.5



### 6.7 46 44 40 37 34 31 29 27 24 22 21 19 18 16 15 14 13 12 11 9.8 9.0



### 6.8 44 41 38 35 32 30 27 25 23 21 20 18 17 15 14 13 12 11 10 9.2 8.5



### 6.9 41 38 35 32 30 28 25 23 21 20 18 17 15 14 13 12 11 10 9.4 8.6 7.9



### 7.0 38 35 33 30 28 25 23 21 20 18 17 15 14 13 12 11 10 9.4 8.6 7.9 7.3



### 7.1 34 32 30 27 25 23 21 20 18 17 15 14 13 12 11 10 9.3 8.5 7.9 7.2 6.7



### 7.2 31 29 27 25 23 21 19 18 16 15 14 13 12 11 9.8 9.1 8.3 7.7 7.1 6.5 6.0



### 7.3 27 26 24 22 20 18 17 16 14 13 12 11 10 9.5 8.7 8.0 7.4 6.8 6.3 5.8 5.3



### 7.4 24 22 21 19 18 16 15 14 13 12 11 9.8 9.0 8.3 7.7 7.0 6.5 6.0 5.5 5.1 4.7



### 7.5 21 19 18 17 15 14 13 12 11 10 9.2 8.5 7.8 7.2 6.6 6.1 5.6 5.2 4.8 4.4 4.0



### 7.6 18 17 15 14 13 12 11 10 9.3 8.6 7.9 7.3 6.7 6.2 5.7 5.2 4.8 4.4 4.1 3.8 3.5



### 7.7 15 14 13 12 11 10 9.3 8.6 7.9 7.3 6.7 6.2 5.7 5.2 4.8 4.4 4.1 3.8 3.5 3.2 2.9



### 7.8 13 12 11 10 9.3 8.5 7.9 7.2 6.7 6.1 5.6 5.2 4.8 4.4 4.0 3.7 3.4 3.2 2.9 2.7 2.5



### 7.9 11 9.9 9.1 8.4 7.7 7.1 6.6 3.0 5.6 5.1 4.7 4.3 4.0 3.7 3.4 3.1 2.9 2.6 2.4 2.2 2.1



### 8.0 8.8 8.2 7.6 7.0 6.4 5.9 5.4 5.0 4.6 4.2 3.9 3.6 3.3 3.0 2.8 2.6 2.4 2.2 2.0 1.9 1.7



### 8.1 7.2 6.8 6.3 5.8 5.3 4.9 4.5 4.1 3.8 3.5 3.2 3.0 2.7 2.5 2.3 2.1 2.0 1.8 1.7 1.5 1.4



### 8.2 6.0 5.6 5.2 4.8 4.4 4.0 3.7 3.4 3.1 2.9 2.7 2.4 2.3 2.1 1.9 1.8 1.6 1.5 1.4 1.3 1.2



### 8.3 4.9 4.6 4.3 3.9 3.6 3.3 3.1 2.8 2.6 2.4 2.2 2.0 1.9 1.7 1.6 1.4 1.3 1.2 1.1 1.0 0.96



### 8.4 4.1 3.8 3.5 3.2 3.0 2.7 2.5 2.3 2.1 2.0 1.8 1.7 1.5 1.4 1.3 1.2 1.1 1.0 0.93 0.86 0.79



### 8.5 3.3 3.1 2.9 2.7 2.4 2.3 2.1 1.9 1.8 1.6 1.5 1.4 1.3 1.2 1.1 0.98 0.90 0.83 0.77 0.71 0.65



### 8.6 2.8 2.6 2.4 2.2 2.0 1.9 1.7 1.6 1.5 1.3 1.2 1.1 1.0 0.96 0.88 0.81 0.75 0.69 0.63 0.58 0.54



### 8.7 2.3 2.2 2.0 1.8 1.7 1.6 1.4 1.3 1.2 1.1 1.0 0.94 0.87 0.80 0.74 0.68 0.62 0.57 0.53 0.49 0.45



### 8.8 1.9 1.8 1.7 1.5 1.4 1.3 1.2 1.1 1.0 0.93 0.86 0.79 0.73 0.67 0.62 0.57 0.52 0.48 0.44 0.41 0.37



### 8.9 1.6 1.5 1.4 1.3 1.2 1.1 1.0 0.93 0.85 0.79 0.72 0.67 0.61 0.56 0.52 0.48 0.44 0.40 0.37 0.34 0.32



### 9.0 1.4 1.3 1.2 1.1 1.0 0.93 0.86 0.79 0.73 0.67 0.62 0.57 0.52 0.48 0.44 0.41 0.37 0.34 0.32 0.29 0.27

## 234

Table N.2. Acute Data W ithout Mussels : Comparison of the F o ur T axa U sed to C alculate the FAV and CM C in the 1999

AWQC and this U pdated 2013 AWQC E xcluding D ata for F reshwater U nionid M ussels

. 1999 Draft Update Acute Criterion (CMC) Magnitude (Salmonids

[ Oncorhync h us spp.] present) 2013 Final Acute Criterion (CMC) Magnitude excluding

Mussels (Salmonids [ Oncorhynchus spp.] absent)

Species GMAV

pH 8.0,

T=25 ° C

(mg TAN/L) 

## GMAV

pH 7.0,

T=20 ° C

(mg TAN/L) 

## Species GMAV

pH 7.0,

T=20 ° C

(mg TAN/L)



## Oncorhynchus sp. (salmonids), includes: O.

aquabonita, O. clarkii, O. gorbuscha, O.

kisutch * , O. mykiss * , and O. tshawytscha * 21.95 99.15 Golden shiner,

Notemigonus crysoleucas 63.02

Orangethroat darter

, Etheostoma spectabile 17.96 74.25 Pebblesnail,

Fluminicola sp. 62.15

Golden shiner,

Notemigonus crysoleucas 14.67 63.02 Lost River sucker,

Deltistes luxatus * 56.62

Mountain whitefish,

Prosopium williamsoni 12.11 51.93 Mountain whitefish,

Prosopium williamsoni 51.93

FAV 11.23 48.21 FAV 76

CMC 5.6 24 CMC 3 8**

* Federally

- listed as endangered or threatened species

** CMC Excluding mussels, with Oncorhynchus present is 24 mg TAN/L to protect the recreationally and commercially important spec ies Rainbow Trout. When

Oncorhynchus is absent, the CMC is based on the mo untain whitefish and is calculated by the ratio of the CMC to the lowest GMAV in the complete acute dataset (0.7249)

times the lowest GMAV in the dataset excluding mussels (51.93 mg TAN/L for mountain whitefish) which results in a CMC of 37.6 5 mg TAN/L at pH 7 and 20 ° C

.

## 235

Table N.3. Temperature an d pH

- Dependent Values of the CMC (Acute Criterion Magnitude )

– Unionid Mussels A bsent and

Oncorhynchus P resent

. Temperature ( ° C)

pH 0

- 14 15 16 17 18 19 20 21 22 23 24 25 26 27 28 29 30

6.5 33 33 33 33 33 33 33 33 33 33 33 33 33 33 31 29 27

6.6 31 31 31 31 31 31 31 31 31 31 31 31 31 31 30 28 26

6.7 30 30 30 30 30 30 30 30 30 30 30 30 30 30 29 26 24

6.8 28 28 28 28 28 28 28 28 28 28 28 28 28 28 27 25 23

6.9 26 26 26 26 26 26 26 26 26 26 26 26 26 26 25 23 21

7.0 24 24 24 24 24 24 24 24 24 24 24 24 24 24 23 21 20

7.1 22 22 22 22 22 22 22 22 22 22 22 22 22 22 21 19 18

7.2 20 20 20 20 20 20 20 20 20 20 20 20 20 20 19 17 16

7.3 18 18 18 18 18 18 18 18 18 18 18 18 18 18 17 16 14

7.4 15 15 15 15 15 15 15 15 15 15 15 15 15 15 15 14 13

7.5 13 13 13 13 13 13 13 13 13 13 13 13 13 13 13 12 11

7.6 11 11 11 11 11 11 11 11 11 11 11 11 11 11 11 10 9.3

7.7 9.6 9.6 9.6 9.6 9.6 9.6 9.6 9.6 9.6 9.6 9.6 9.6 9.6 9.6 9.3 8.6 7.9

7.8 8.1 8.1 8.1 8.1 8.1 8.1 8.1 8.1 8.1 8.1 8.1 8.1 8.1 8.1 7.8 7.2 6.6

7.9 6.8 6.8 6.8 6.8 6.8 6.8 6.8 6.8 6.8 6.8 6.8 6.8 6.8 6.8 6.5 6.0 5.5

8.0 5.6 5.6 5.6 5.6 5.6 5.6 5.6 5.6 5.6 5.6 5.6 5.6 5.6 5.6 5.4 5.0 4.6

8.1 4.6 4.6 4.6 4.6 4.6 4.6 4.6 4.6 4.6 4.6 4.6 4.6 4.6 4.6 4.5 4.1 3.8

8.2 3.8 3.8 3.8 3.8 3.8 3.8 3.8 3.8 3.8 3.8 3.8 3.8 3.8 3.8 3.7 3.4 3.1

8.3 3.2 3.2 3.2 3.2 3.2 3.2 3.2 3.2 3.2 3.2 3.2 3.2 3.2 3.2 3.0 2.8 2.6

8.4 2.6 2.6 2.6 2.6 2.6 2.6 2.6 2.6 2.6 2.6 2.6 2.6 2.6 2.6 2.5 2.3 2.1

8.5 2.1 2.1 2.1 2.1 2.1 2.1 2.1 2.1 2.1 2.1 2.1 2.1 2.1 2.1 2.1 1.9 1.8

8.6 1.8 1.8 1.8 1.8 1.8 1.8 1.8 1.8 1.8 1.8 1.8 1.8 1.8 1.8 1.7 1.6 1.4

8.7 1.5 1.5 1.5 1.5 1.5 1.5 1.5 1.5 1.5 1.5 1.5 1.5 1.5 1.5 1.4 1.3 1.2

8.8 1.2 1.2 1.2 1.2 1.2 1.2 1.2 1.2 1.2 1.2 1.2 1.2 1.2 1.2 1.2 1.1 1.0

8.9 1.0 1.0 1.0 1.0 1.0 1.0 1.0 1.0 1.0 1.0 1.0 1.0 1.0 1.0 1.0 0.92 0.85

9.0 0.88 0.88 0.88 0.88 0.88 0.88 0.88 0.88 0.88 0.88 0.88 0.88 0.88 0.88 0.85 0.78 0.72

## 236

Table N.4. Temperature an d pH

- Dependent Values of the CMC (Acute Criterion Magnitude )

– Unionid Mussels A bsent and

Oncorhynchus A bsent

. Temperature ( ° C)

pH 0

- 14 15 16 17 18 19 20 21 22 23 24 25 26 27 28 29 30

6.5 51 51 51 51 51 51 51 51 51 48 44 40 37 34 31 29 27

6.6 49 49 49 49 49 49 49 49 49 46 42 39 36 33 30 28 26

6.7 46 46 46 46 46 46 46 46 46 43 40 37 34 31 29 26 24

6.8 44 44 44 44 44 44 44 44 44 41 38 35 32 29 27 25 23

6.9 41 41 41 41 41 41 41 41 41 38 35 32 30 27 25 23 21

7.0 38 38 38 38 38 38 38 38 38 35 32 30 27 25 23 21 20

7.1 34 34 34 34 34 34 34 34 34 32 29 27 25 23 21 19 18

7.2 31 31 31 31 31 31 31 31 31 29 26 24 22 21 19 17 16

7.3 27 27 27 27 27 27 27 27 27 26 23 22 20 18 17 16 14

7.4 24 24 24 24 24 24 24 24 24 22 21 19 17 16 15 14 13

7.5 21 21 21 21 21 21 21 21 21 19 18 16 15 14 13 12 11

7.6 18 18 18 18 18 18 18 18 18 17 15 14 13 12 11 10 9.3

7.7 15 15 15 15 15 15 15 15 15 14 13 12 11 10 9.3 8.6 7.9

7.8 13 13 13 13 13 13 13 13 13 12 11 10 9.2 8.5 7.8 7.2 6.6

7.9 11 11 11 11 11 11 11 11 11 9.9 9.1 8.4 7.7 7.1 6.5 6.0 5.5

8.0 8.8 8.8 8.8 8.8 8.8 8.8 8.8 8.8 8.8 8.2 7.5 6.9 6.4 5.9 5.4 5.0 4.6

8.1 7.3 7.3 7.3 7.3 7.3 7.3 7.3 7.3 7.3 6.8 6.2 5.7 5.3 4.9 4.5 4.1 3.8

8.2 6.0 6.0 6.0 6.0 6.0 6.0 6.0 6.0 6.0 5.6 5.1 4.7 4.4 4.0 3.7 3.4 3.1

8.3 4.9 4.9 4.9 4.9 4.9 4.9 4.9 4.9 4.9 4.6 4.2 3.9 3.6 3.3 3.0 2.8 2.6

8.4 4.1 4.1 4.1 4.1 4.1 4.1 4.1 4.1 4.1 3.8 3.4 3.2 3.0 2.7 2.5 2.3 2.1

8.5 3.3 3.3 3.3 3.3 3.3 3.3 3.3 3.3 3.3 3.1 2.9 2.6 2.4 2.2 2.1 1.9 1.8

8.6 2.8 2.8 2.8 2.8 2.8 2.8 2.8 2.8 2.8 2.6 2.4 2.2 2.0 1.9 1.7 1.6 1.4

8.7 2.3 2.3 2.3 2.3 2.3 2.3 2.3 2.3 2.3 2.2 2.0 1.8 1.7 1.5 1.4 1.3 1.2

8.8 1.9 1.9 1.9 1.9 1.9 1.9 1.9 1.9 1.9 1.8 1.7 1.5 1.4 1.3 1.2 1.1 1.0

8.9 1.6 1.6 1.6 1.6 1.6 1.6 1.6 1.6 1.6 1.5 1.4 1.3 1.2 1.1 1.0 0.92 0.85

9.0 1.4 1.4 1.4 1.4 1.4 1.4 1.4 1.4 1.4 1.3 1.2 1.1 1.0 0.93 0.85 0.78 0.72

## 237

Table N.5

. 2013 Acute Criterion Recalculations for Site

- specific Criteria

. Acute Criterion Duration

(1 hr average)

at pH 7 and 20 ° C

(mg TAN/L) Acute Criterion Magnitude

(CMC)

Oncorhynchus sp p.

(Rainbow Trout ) P resent Acute Criterion Magnitude

(CMC)

Oncorhynchus sp p.

(Rainbow Trout) A bsent

Mussels P resent 17 17

Mussels A bsent 24 38

Frequency: Criteria values not to be exceeded more than once in three years.

Table N.6 . Chronic Dataset W ithout Mussels : Comparison of the F o ur T axa used to

Calculate the CCC in the 1999 AWQC and this U pdated 2013 AWQC E xcluding D ata for

F reshwater U nionid M ussels

.

1999 Draft Update Chronic Criterion (CCC) Magnitude 2013 Final Chronic Criterion (CCC)

Magnitude excluding mussels

Species GMCV

pH 8.0,

T=25 ° C

( mg TAN/L ) GMCV

pH 7.0,

T=20 ° C

( mg TAN/L ) Species GMCV

pH 7.0,

T=20 ° C

( mg TAN/L )

Fathead minnow,

Pimephales promelas 3.09 7.503 Fathead minnow,

Pimephales promelas 9.187

Lepomis sp.

(Centrarchidae), includes:

Bluegill sunfish, L.

macrochirus , and Green

sunfish, L. cyanellus 2.85 6.92 Pebblesnail,

Fluminicola sp. 7.828

Long fingernail clam,

Musculium transversum <2.26 7.547 Long fingernail clam,

Musculium transversum 7.547

Amphipod,

Hyalella azteca <1.45 4.865 Lepomis sp.

(Centrarchidae), includes:

Bluegill, L. macrochirus and

Green sunfish , L. cyanellus 6.92 0

CCC 1.2 4.5 * CCC 6.5

*Based on data renormalized to pH 7.0 and T 20 ° C

## 238

Table N

. 7

. Chronic Criterion Recalculations for Site

- S pecific Criteria

. Chronic Criterion Duration

(30

- day average)

at pH 7 and 20 ° C

(mg TAN/L) Chronic Criterion

Magnitude (CCC)

Fish ELS P resent Chronic Criterion

Magnitude (CCC)

Fish ELS A bsent

Mussels P resent 1.9 1.9

Mussels A bsent 6.5 7.1

Not to exceed 2.5 times the CCC as a 4

- day average within the 30

- day averaging period.

Frequency: Criteria values not to be exceeded more than once in three years.

## 239

Table N.8. Temperature an d pH

- Dependent Values of the CCC (Chronic Criterion Magnitude )

– Mussels A bsent and E arly

L ife S tage (ELS) P rotection N ecessary

. Temperature ( ° C)

pH 0

- 14 15 16 17 18 19 20 21 22 23 24 25 26 27 28 29 30

6.5 7.3 7.3 7.3 7.3 7.3 7.3 7.3 7.3 7.0 6.6 6.2 5.8 5.4 5.1 4.8 4.5 4.2

6.6 7.2 7.2 7.2 7.2 7.2 7.2 7.2 7.2 6.9 6.5 6.1 5.7 5.4 5.0 4.7 4.4 4.1

6.7 7.1 7.1 7.1 7.1 7.1 7.1 7.1 7.1 6.8 6.4 6.0 5.6 5.3 4.9 4.6 4.3 4.1

6.8 6.9 6.9 6.9 6.9 6.9 6.9 6.9 6.9 6.6 6.2 5.8 5.5 5.1 4.8 4.5 4.2 4.0

6.9 6.7 6.7 6.7 6.7 6.7 6.7 6.7 6.7 6.5 6.1 5.7 5.3 5.0 4.7 4.4 4.1 3.9

7.0 6.5 6.5 6.5 6.5 6.5 6.5 6.5 6.5 6.2 5.8 5.5 5.1 4.8 4.5 4.2 4.0 3.7

7.1 6.2 6.2 6.2 6.2 6.2 6.2 6.2 6.2 6.0 5.6 5.3 4.9 4.6 4.3 4.1 3.8 3.6

7.2 5.9 5.9 5.9 5.9 5.9 5.9 5.9 5.9 5.7 5.3 5.0 4.7 4.4 4.1 3.9 3.6 3.4

7.3 5.6 5.6 5.6 5.6 5.6 5.6 5.6 5.6 5.4 5.0 4.7 4.4 4.1 3.9 3.6 3.4 3.2

7.4 5.2 5.2 5.2 5.2 5.2 5.2 5.2 5.2 5.0 4.7 4.4 4.1 3.9 3.6 3.4 3.2 3.0

7.5 4.8 4.8 4.8 4.8 4.8 4.8 4.8 4.8 4.6 4.3 4.1 3.8 3.6 3.3 3.1 2.9 2.8

7.6 4.4 4.4 4.4 4.4 4.4 4.4 4.4 4.4 4.2 3.9 3.7 3.5 3.2 3.0 2.9 2.7 2.5

7.7 3.9 3.9 3.9 3.9 3.9 3.9 3.9 3.9 3.8 3.5 3.3 3.1 2.9 2.7 2.6 2.4 2.3

7.8 3.5 3.5 3.5 3.5 3.5 3.5 3.5 3.5 3.4 3.2 3.0 2.8 2.6 2.4 2.3 2.1 2.0

7.9 3.1 3.1 3.1 3.1 3.1 3.1 3.1 3.1 3.0 2.8 2.6 2.4 2.3 2.1 2.0 1.9 1.8

8.0 2.7 2.7 2.7 2.7 2.7 2.7 2.7 2.7 2.6 2.4 2.3 2.1 2.0 1.9 1.7 1.6 1.5

8.1 2.3 2.3 2.3 2.3 2.3 2.3 2.3 2.3 2.2 2.1 1.9 1.8 1.7 1.6 1.5 1.4 1.3

8.2 2.0 2.0 2.0 2.0 2.0 2.0 2.0 2.0 1.9 1.8 1.7 1.6 1.5 1.4 1.3 1.2 1.1

8.3 1.7 1.7 1.7 1.7 1.7 1.7 1.7 1.7 1.6 1.5 1.4 1.3 1.2 1.2 1.1 1.0 0.96

8.4 1.4 1.4 1.4 1.4 1.4 1.4 1.4 1.4 1.4 1.3 1.2 1.1 1.1 0.99 0.93 0.87 0.81

8.5 1.2 1.2 1.2 1.2 1.2 1.2 1.2 1.2 1.2 1.1 1.0 0.95 0.89 0.83 0.78 0.73 0.69

8.6 1.0 1.0 1.0 1.0 1.0 1.0 1.0 1.0 0.97 0.91 0.85 0.80 0.75 0.70 0.66 0.62 0.58

8.7 0.86 0.86 0.86 0.86 0.86 0.86 0.86 0.86 0.82 0.77 0.72 0.68 0.64 0.60 0.56 0.52 0.49

8.8 0.73 0.73 0.73 0.73 0.73 0.73 0.73 0.73 0.70 0.65 0.61 0.58 0.54 0.51 0.47 0.44 0.42

8.9 0.62 0.62 0.62 0.62 0.62 0.62 0.62 0.62 0.60 0.56 0.52 0.49 0.46 0.43 0.41 0.38 0.36

9.0 0.54 0.54 0.54 0.54 0.54 0.54 0.54 0.54 0.51 0.48 0.45 0.42 0.40 0.37 0.35 0.33 0.31

## 240

Table N.9. Temperature an d pH

- Dependent Values of the CCC (Chronic Criterion Magnitude )

– Mussels A bsent and E arly

L ife S tage (ELS) P rotection not N ecessary

. Temperatur e ( 

## ° C)



## pH 0

- 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25 26 27 28 29 30

6.5 19 17 16 15 14 13 13 12 11 10 9.7 9.1 8.5 8.0 7.5 7.0 6.6 6.2 5.8 5.4 5.1 4.8 4.5 4.2



### 6.6 18 17 16 15 14 13 12 12 11 10 9.6 9.0 8.4 7.9 7.4 6.9 6.5 6.1 5.7 5.4 5.0 4.7 4.4 4.1



### 6.7 18 17 16 15 14 13 12 11 11 10 9.4 8.8 8.3 7.7 7.3 6.8 6.4 6.0 5.6 5.3 4.9 4.6 4.3 4.1



### 6.8 17 16 15 14 14 13 12 11 10 9.8 9.2 8.6 8.1 7.6 7.1 6.7 6.2 5.8 5.5 5.1 4.8 4.5 4.2 4.0



### 6.9 17 16 15 14 13 12 12 11 10 9.5 8.9 8.4 7.8 7.4 6.9 6.5 6.1 5.7 5.3 5.0 4.7 4.4 4.1 3.9



### 7.0 16 15 14 14 13 12 11 10 9.8 9.2 8.6 8.1 7.6 7.1 6.7 6.2 5.9 5.5 5.1 4.8 4.5 4.2 4.0 3.7



### 7.1 16 15 14 13 12 11 11 10 9.4 8.8 8.3 7.7 7.3 6.8 6.4 6.0 5.6 5.3 4.9 4.6 4.3 4.1 3.8 3.6



### 7.2 15 14 13 12 12 11 10 9.5 9.0 8.4 7.9 7.4 6.9 6.5 6.1 5.7 5.3 5.0 4.7 4.4 4.1 3.9 3.6 3.4



### 7.3 14 13 12 12 11 10 9.6 9.0 8.4 7.9 7.4 6.9 6.5 6.1 5.7 5.4 5.0 4.7 4.4 4.1 3.9 3.6 3.4 3.2



### 7.4 13 12 12 11 10 9.5 9.0 8.4 7.9 7.4 6.9 6.5 6.1 5.7 5.3 5.0 4.7 4.4 4.1 3.9 3.6 3.4 3.2 3.0



### 7.5 12 11 11 10 9.4 8.8 8.2 7.7 7.2 6.8 6.4 6.0 5.6 5.2 4.9 4.6 4.3 4.1 3.8 3.6 3.3 3.1 2.9 2.8



### 7.6 11 10 10 9.1 8.5 8.0 7.5 7.0 6.6 6.2 5.8 5.4 5.1 4.8 4.5 4.2 3.9 3.7 3.5 3.2 3.0 2.9 2.7 2.5



### 7.7 9.9 9.3 8.7 8.1 7.7 7.2 6.8 6.3 5.9 5.6 5.2 4.9 4.6 4.3 4.0 3.8 3.5 3.3 3.1 2.9 2.7 2.6 2.4 2.3



### 7.8 8.8 8.3 7.8 7.3 6.8 6.4 6.0 5.6 5.3 5.0 4.6 4.4 4.1 3.8 3.6 3.4 3.2 3.0 2.8 2.6 2.4 2.3 2.1 2.0



### 7.9 7.8 7.3 6.8 6.4 6.0 5.6 5.3 5.0 4.6 4.4 4.1 3.8 3.6 3.4 3.2 3.0 2.8 2.6 2.4 2.3 2.1 2.0 1.9 1.8



### 8.0 6.8 6.3 6.0 5.6 5.2 4.9 4.6 4.3 4.0 3.8 3.6 3.3 3.1 2.9 2.7 2.6 2.4 2.3 2.1 2.0 1.9 1.7 1.6 1.5



### 8.1 5.8 5.5 5.1 4.8 4.5 4.2 4.0 3.7 3.5 3.3 3.1 2.9 2.7 2.5 2.4 2.2 2.1 2.0 1.8 1.7 1.6 1.5 1.4 1.3



### 8.2 5.0 4.7 4.4 4.1 3.9 3.6 3.4 3.2 3.0 2.8 2.6 2.5 2.3 2.2 2.0 1.9 1.8 1.7 1.6 1.5 1.4 1.3 1.2 1.1



### 8.3 4.2 4.0 3.7 3.5 3.3 3.1 2.9 2.7 2.5 2.4 2.2 2.1 2.0 1.8 1.7 1.6 1.5 1.4 1.3 1.2 1.2 1.1 1.0 0.96



### 8.4 3.6 3.4 3.2 3.0 2.8 2.6 2.4 2.3 2.1 2.0 1.9 1.8 1.7 1.6 1.5 1.4 1.3 1.2 1.1 1.1 0.99 0.92 0.87 0.81



### 8.5 3.0 2.8 2.7 2.5 2.3 2.2 2.1 1.9 1.8 1.7 1.6 1.5 1.4 1.3 1.2 1.2 1.1 1.0 0.95 0.89 0.83

3 0.78 0.73 0.69



### 8.6 2.6 2.4 2.2 2.1 2.0 1.9 1.7 1.6 1.5 1.4 1.3 1.3 1.2 1.1 1.0 0.97 0.91 0.85 0.80 0.75 0.70 0.66 0.62 0.58



### 8.7 2.2 2.0 1.9 1.8 1.7 1.6 1.5 1.4 1.3 1.2 1.1 1.1 1.0 0.93 0.88 0.82 0.77 0.72 0.68 0.63 0.60 0.56 0.52 0.49



### 8.8 1.8 1.7 1.6 1.5 1.4 1.3 1.3 1.2 1.1 1.0 0.96 0.90 0.85 0.79 0.74 0.70 0.65 0.61 0.58 0.54 0.51 0.47 0.44 0.42



### 8.9 1.6 1.5 1.4 1.3 1.2 1.1 1.1 1.0 0.94 0.88 0.82 0.77 0.72 0.68 0.64 0.60 0.56 0.52 0.49 0.46 0.43 0.40 0.38 0.36



### 9.0 1.4 1.3 1.2 1.1 1.0 0.98 0.92 0.86

6 0.81 0.76 0.71 0.66 0.62 0.58 0.55 0.51 0.48 0.45 0.42 0.40 0.37 0.35 0.33 0.31

## 241

Figure N.1. Comparison of the 2013 CMC Extrapolated A cross a T emperature G radient

at pH 7 A ccounting for the P resence or A bsence of U nionid M ussels and the P resence or

A bsence of Oncorhynchus

.

## 242

Figure N.2. Comparison of the 2013 CCC E xtrapolated A cross a T emperature G radient at

pH 7 A ccounting for the P resence or A bsence of M ussels and/or the N eed for E arly L ife

S tage (ELS) P rotection of F ish S pecies.