EPA is proposing to make corrections and clarifications to the following three Whole Effluent Toxicity (WET) methods manuals: 

Methods for Measuring the Acute Toxicity of Effluents and Receiving Waters to Freshwater and Marine Organisms, Fifth Edition. U.S. Environmental Protection Agency, Office 

of Water, Washington, DC EPA-821-R-02-012, October 2002. Short title: Acute Freshwater and Marine Manual.

Short-term Methods for Estimating the Chronic Toxicity of Effluents and Receiving Waters to Freshwater Organisms, Fourth Edition. U.S. Environmental Protection Agency, 

Office of Water, Washington, DC EPA/821/R–02/013, October 2002. Short title: Chronic Freshwater Manual.

Methods for Measuring the Chronic Toxicity of Effluents and Receiving Waters to Marine and Estuarine Organisms, Third Edition. U.S. Environmental Protection Agency, Office 

of Water, Washington DC EPA/821/R–02/014, October 2002. Short title: Chronic Marine and Estuarine Manual.

Three tables (one for each document in the above order) are included in this document. Each table specifies: 

1. Where in the document the proposed change occurs

2. How the current text reads

3. What correction is being made

4. How the corrected text will read

5. The reason for the correction

WET Methods Errata Sheet 1 October 2014
Errata for Methods for Measuring the Acute Toxicity of Effluents and Receiving Waters to Freshwater and Marine Organisms, Fifth Edition. U.S. Environmental 

Protection Agency, Office of Water, Washington, DC EPA-821-R-02-012, October 2002

Text Location Current Text Correction Being Made Corrected Text Reason For Correction 

Section 4, Quality 

Assurance, Section 

4.4.1, Laboratory Water 

Used for Culturing and 

Test Dilution Water, 

Page 8

Water used for culturing and test 

dilution should be analyzed for 

toxic metals and organics at least 

annually or whenever difficulty is 

encountered in meeting minimum 

acceptability criteria for control 

survival and reproduction or 

growth.

Add "test" between 

"minimum" and 

"acceptability criteria." 

Water used for culturing and test dilution should be 

analyzed for toxic metals and organics at least annually 

or whenever difficulty is encountered in meeting 

minimum test acceptability criteria for control survival 

and reproduction or growth.

Clarification.

Section 4, Quality 

Assurance, Section 

4.8.3, Food Quality, 

Page 9

For foods (e.g., such as YCT) 

which are used to culture and test 

organisms, the quality of food 

should meet the requirements for 

the laboratory water used for 

culturing and test dilution water as 

described in Section 4.4 above.

Spell out acronym for 

YCT. Add "at the 

concentration in the 

culture or test solutions" at 

the end of the sentence. 

For foods (e.g., such as Yeast, Cereal leaves, and Trout 

chow (YCT)) which are used to culture and test 

organisms, the quality of food should meet the 

requirements for the laboratory water used for culturing 

and test dilution water as described in Section 4.4 above 

at the concentration in the culture or test solutions.

Clarification. Define the 

YCT components. Analysis 

should be at the 

concentration in the culture 

or in the test solutions so that 

the foods meet the 

requirements at the feeding 

concentrations.

Section 4, Quality 

Assurance, Section 

4.15.2, Documenting 

Ongoing Laboratory 

Performance, Page 18 

A control chart should be prepared 

for each combination of reference 

toxicant, test species, test 

condition, and endpoint. 

Change singular words 

"condition" and 

"endpoint" to plural.

A control chart should be prepared for each combination 

of reference toxicant, test species, test conditions, and 

endpoints.

Consistency correction with 

Section 4.16.2 in "Chronic 

Freshwater Manual" and 

"Chronic Marine and 

Estuarine Manual." 

Section 4, Quality 

Assurance, Section 

4.15.2, Documenting 

Ongoing Laboratory 

Performance, Page 18

... for establishing the control 

charts. In this technique, a 

running plot is maintained for the 

toxicity values (Xi) from 

successive tests with a given 

reference toxicant (Figure 1), and 

endpoints (LC50s) are examined 

to determine if they are within the 

prescribed limits. The types of 

control charts illustrated.... 

Replace sentence "In this 

technique..." with two 

sentences – "Successive 

toxicity endpoints 

(NOECs, IC25s, LC50s, 

etc.) should be plotted and 

examined to determine if 

the results (X1) are within 

prescribed limits (Figure 

1). The chart should plot 

logarithm of concentration 

on the vertical axis against 

the date of the test or test 

number on the horizontal 

axis."

... for establishing the control charts. Successive 

toxicity endpoints (NOECs, IC25s, LC50s, etc.) should 

be plotted and examined to determine if the results (X1) 

are within prescribed limits (Figure 1). The chart should 

plot logarithm of concentration on the vertical axis 

against the date of the test or test number on the 

horizontal axis. The types of control charts illustrated.... 

Consistency correction with 

Section 4.16.2 in "Chronic 

Freshwater Manual" and 

"Chronic Marine and 

Estuarine Manual."

WET Methods Errata Sheet 2 October 2014
Errata for Methods for Measuring the Acute Toxicity of Effluents and Receiving Waters to Freshwater and Marine Organisms, Fifth Edition. U.S. Environmental 

Protection Agency, Office of Water, Washington, DC EPA-821-R-02-012, October 2002

Text Location Current Text Correction Being Made Corrected Text Reason For Correction 

Section 4, Quality 

Assurance, Section 

4.15.2, Figure 1, 

Documenting Ongoing

Laboratory Performance, 

Page 20

Replace the graphs in 

Figure 1 with log scale 

graphs. Add explanatory 

text to Figure 1 legend.

Figure 1. Control (cusum) charts: A. General case; B 

and C, 48-h acute tests with sodium chloride, (B) 

Fathead minnow (Pimephales promelas), and (C)

Ceriodaphnia dubia. Charts are prepared in log format 

with individual LC50 data plotted on a graph. Note 

Figures B and C are plotted using anti-logarithms. The 

mean of the logarithmic LC is the central tendency. The 

upper and lower control limits are calculated as the 

Correction and clarification. 

Cusum charting can either 

plot anti-logarithm values for 

historical mean and warning 

limits on the graph with the 

y-axis scaled as logarithm or 

plot logarithm values for 

historical mean and warning 

limits on the graph with the 

y-axis as normal arithmetic 

scaling. The figures B and C 

are revised to match the text 

in the manual and are shown 

using a log format with the 

individual LC or EC data 

plotted on each graph. 

Cusum charts may be plotted 

using antilogarithm values so 

that the original units (e.g., 

g/L) are displayed on the 

graph. The log values on a 

log scaled y-axis or antilog 

values on an arithmetic scale 

y-axis are not be mixed on a 

chart. Note Figures B and C 

are redrawn for these graphs 

using the antilog values of 

the LC50 as many graphics 

programs do not allow 

enlarging the log scale axis 

sufficiently (i.e., expanding 

within a log unit), and trends 

can be difficult to view; 

therefore graphs using 

antilogarithm values may be

preferable. Reference: 

Environment Canada. 2009. 

Environment Canada’s 

Guidance on Using 

Logarithms for Reference 

Toxicant Warning Chart. 

Biological Methods Section, 

September.

WET Methods Errata Sheet 3 October 2014
Errata for Methods for Measuring the Acute Toxicity of Effluents and Receiving Waters to Freshwater and Marine Organisms, Fifth Edition. U.S. Environmental 

Protection Agency, Office of Water, Washington, DC EPA-821-R-02-012, October 2002

Text Location Current Text Correction Being Made Corrected Text Reason For Correction 

average LC50 value (± 2S) and plotted on the chart as 

horizontal lines serving as visual indicators of any trend 

of successive effect endpoints that are divergent results. 

Cusum charting can plot logarithm values for historical 

mean and warning limits on the graph with the y-axis as 

normal arithmetic scaling or plot LC50 values for 

historical mean and warning limits on the graph with the 

y-axis scaled as logarithm. Antilogarithm values can be 

used so that the original units (e.g., g/L) are displayed on 

the graph. The log values on a log scaled y-axis or 

antilog values on an arithmetic scale y-axis are not be 

mixed on a chart. (Data provided by the Environmental 

Services Division, U.S. Environmental Protection 

Agency, Kansas City, KS).

Section 4, Quality 

Assurance, Section 

4.16.1, Reference 

Toxicants, Page 21

...requiring aquatic toxicity tests. 

EMSL-Cincinnati hopes to release 

EPA-certified solutions of 

cadmium and copper with 

accompanying toxicity data for the 

recommended test species, for use 

as reference toxicants through 

cooperative research and 

development agreements with 

commercial suppliers, and will 

continue to develop additional 

reference toxicants for future 

release. Standard reference 

materials....

Delete sentence "EMSLCincinnati hopes to...." ...requiring aquatic toxicity tests. Standard reference 

materials.... 

EMSL-Cincinnati no longer 

provides reference toxicants. 

Section 5, Facilities and 

Equipment, Section 5.4, 

Reagents and 

Consumable Materials, 

Section 5.4.9.4, Dried, 

Powdered Leaves 

(CEROPHYLL®), Page 

25

Dried, Powdered Leaves 

(CEROPHYLL®). Dried, 

powdered, cereal leaves (e.g., 

CEROPHYLL® or equivalent) are 

available....

Delete term 

CEROPHYLL® from 

section title. Delete "(e.g., 

CEROPHYLL® or 

equivalent)" from first 

sentence.

Dried, Powdered Leaves. Dried, powdered, cereal leaves 

are available.... 

CEROPHYLL®, a 

commercial product, is no 

longer available.

WET Methods Errata Sheet 4 October 2014
Errata for Methods for Measuring the Acute Toxicity of Effluents and Receiving Waters to Freshwater and Marine Organisms, Fifth Edition. U.S. Environmental 

Protection Agency, Office of Water, Washington, DC EPA-821-R-02-012, October 2002

Text Location Current Text Correction Being Made Corrected Text Reason For Correction 

Section 6, Test 

Organisms, Section 

6.6.4, Transportation to 

the Test Site, Page 30

In static tests, marine organisms 

can be used at all concentrations 

of effluent by adjusting the 

salinity of the effluent to a 

standard salinity (such as 25‰) or 

to the salinity approximating that 

of the receiving water, by adding 

sufficient dry ocean salts, such as 

Forty Fathoms®, or equivalent, 

GP2 or hypersaline brine.

Replace "Forty Fathoms®"

with "HW 

MARINEMIX®" in 

sentence.

...by adding sufficient dry ocean salts, such as HW 

MARINEMIX®, or equivalent.... 

FORTY FATHOMS®, a 

commercial product, is no 

longer available. Equivalent 

commercial products, such as 

HW MARINEMIX®, are 

acceptable replacements. 

Section 7, Dilution 

Water, Section 7.2.1, 

Standard, Synthetic 

Dilution Water, Page 31 

Standard, synthetic, dilution water 

is prepared with deionized water 

and reagent grade chemicals or 

mineral water (Tables 7 and 8) 

and commercial sea salts (FORTY 

FATHOMS®, HW 

MARINEMIX®) (Table 9). 

Delete term "FORTY 

FATHOMS®" and add "or 

equivalent" to sentence.

...and commercial salts (HW MARINEMIX® or 

equivalent) (Table 9). 

FORTY FATHOMS®, a 

commercial product, is no 

longer available. Equivalent 

commercial products, such as 

HW MARINEMIX®, are 

acceptable replacements.

Section 7, Dilution 

Water, Section 7.2.4.2, 

Standard, Synthetic 

Seawater, Page 33

Synthetic seawater can also be 

prepared by adding commercial 

sea salts, such as FORTY 

FATHOMS®, HW 

MARINEMIX® or equivalent, to 

deionized water. For example, 

thirty-one parts per thousand 

(31‰) FORTY FATHOMS® can 

be prepared by dissolving 31 g of 

product per liter of deionized 

water. The salinity of the 

resulting solutions should be 

checked with a refractometer.

Delete "FORTY 

FATHOMS®" from the 

first sentence. Delete 

second sentence.

...can also be prepared by adding commercial sea salts, 

such as HW MARINEMIX® or equivalent to deionized 

water. The salinity of the resulting solutions....

FORTY FATHOMS®, a 

commercial product, is no 

longer available. Equivalent 

commercial products, such as 

HW MARINEMIX®, are 

acceptable replacements.

Section 7, Dilution 

Water, Section 7.3.4, 

Use of Receiving Water 

as Dilution Water, Page 

33

Where necessary, the salinity of a 

surface water can be increased by 

the addition of artificial sea salts, 

such as FORTY FATHOMS® or 

equivalent, a natural seawater of 

higher salinity, or hypersaline 

brine.

Replace "Forty Fathoms®"

with "HW 

MARINEMIX®" in 

sentence.

...addition of artificial sea salts, HW MARINEMIX®, or 

equivalent....

FORTY FATHOMS®, a 

commercial product, is no 

longer available. Equivalent 

commercial products, such as 

HW MARINEMIX®, are 

acceptable replacements.

WET Methods Errata Sheet 5 October 2014
Errata for Methods for Measuring the Acute Toxicity of Effluents and Receiving Waters to Freshwater and Marine Organisms, Fifth Edition. U.S. Environmental 

Protection Agency, Office of Water, Washington, DC EPA-821-R-02-012, October 2002

Text Location Current Text Correction Being Made Corrected Text Reason For Correction 

Section 9, Acute 

Toxicity Test 

Procedures, Section

9.5.3, Static Tests, Page 

44

Saline dilution water can be 

prepared by adding dry salts 

(FORTY FATHOMS® or 

equivalent, or modified GP2) or 

hypersaline brine to de-ionized 

water, or a suitable surface 

freshwater, to adjust the salinity of 

the entire dilution series.

Replace "Forty Fathoms®"

with "HW 

MARINEMIX®" and 

delete "modified" in 

sentence.

...adding dry salts (HW MARINEMIX® or equivalent, or 

GP2) or....

FORTY FATHOMS®, a 

commercial product, is no 

longer available. Equivalent 

commercial products, such as 

HW MARINEMIX®, are 

acceptable replacements.

Section 9, Acute 

Toxicity Test 

Procedures, Section 

9.11.1, Feeding, Page 46

The organisms are fed at test

renewal, 48 h after the test is 

initiated, if Regional or State 

policy requires a 96-h test 

duration. 

Replace "fed at test 

renewal, 48 h after the test

is initiated" with "are 

provided food 2 h prior to 

test renewal at 48h." 

The organisms are provided food 2 h prior to test 

solution renewal at 48 h, if Regional or State policy 

requires a 96 h test duration. 

Clarification of when foods 

can be added to match the 

text in the tables. The feeding 

regimes require that food(s) 

be made available while 

holding prior to the test; for 

some methods (Table 12, 13) 

for a minimum of 2 h prior 

renewing the test solutions in 

a test. Any feeding regimes 

require that the food is added 

2 h prior to test solution 

renewal at 48 h 

(recommended); except for 

the mysid test, where food is 

added daily. 

Section 9, Acute 

Toxicity Test 

Procedures, Section 

9.11.2, Feeding, Page 46 

Where Artemia nauplii are fed, the 

nauplii are first concentrated...." 

Add "the < 24 h old" after 

comma in sentence.

Where Artemia nauplii are fed, the ≤ 24 h old nauplii are 

first concentrated ... 

Clarification of age of the 

brine shrimp to be fed. 

Section 9, Acute 

Toxicity Test 

Procedures, Section 

9.171, Summary of Test 

Conditions and Test 

Acceptability Criteria for 

Mysid, Mysidopsis 

bahia, Acute Toxicity 

Tests With Effluents and 

Receiving Waters (Test 

Method 2007.0) 

(Continued), Table 16, 

Line 17, Dilution Water, 

Page 60

5-30‰ ±10%; Uncontaminated 

source of seawater, deionized 

water mixed with hypersaline 

brine or artificial sea salts (HW 

MARINEMIX®, FORTY 

FATHOMS®, modified GP2, or 

equivalent) prepared with MILLIQ® or equivalent deionized water 

(see Section 7, Dilution Water); or 

receiving water (available options)

Delete "FORTY 

FATHOMS®," from 

sentence.

...or artificial sea salts (HW MARINEMIX®, modified 

GP2, or equivalent)....

FORTY FATHOMS®, a 

commercial product, is no 

longer available. Equivalent 

commercial products, such as 

HW MARINEMIX®, are 

acceptable replacements.

WET Methods Errata Sheet 6 October 2014
Errata for Methods for Measuring the Acute Toxicity of Effluents and Receiving Waters to Freshwater and Marine Organisms, Fifth Edition. U.S. Environmental 

Protection Agency, Office of Water, Washington, DC EPA-821-R-02-012, October 2002

Text Location Current Text Correction Being Made Corrected Text Reason For Correction 

Section 9, Acute 

Toxicity Test 

Procedures, Section 

9.171, Summary of Test 

Conditions and Test 

Acceptability Criteria for 

Sheepshead Minnows, 

Cyprinodon variegatus, 

Acute Toxicity Tests 

With Effluents and 

Receiving Waters (Test 

Method 2004.0) 

(Continued), Table 17, 

Line 17, Dilution Water, 

Page 62

5-32‰ ±10%; Uncontaminated 

source of seawater, deionized 

water mixed with hypersaline 

brine or artificial sea salts (HW 

MARINEMIX®, FORTY 

FATHOMS®, modified GP2, or 

equivalent) prepared with MILLIQ® or equivalent deionized water 

(see Section 7, Dilution Water); or 

receiving water (available options)

Delete "FORTY 

FATHOMS®," from 

sentence.

...or artificial sea salts (HW MARINEMIX®, modified 

GP2, or equivalent)....

FORTY FATHOMS®, a 

commercial product, is no 

longer available. Equivalent 

commercial products, such as 

HW MARINEMIX®, are 

acceptable replacements.

Section 9, Acute 

Toxicity Test 

Procedures, Section 

9.171, Summary of Test 

Conditions and Test 

Acceptability Criteria for 

Silverside, Menidia 

beryllina, M. menidia, 

and M. peninsulae, 

Acute Toxicity Tests 

With Effluents and 

Receiving Waters (Test 

Method 2006.0) 

(Continued), Table 18, 

Line 17, Dilution Water, 

Page 64

5-32‰ ±10%; Uncontaminated 

source of seawater, deionized 

water mixed with hypersaline 

brine or artificial sea salts (HW 

MARINEMIX®, FORTY 

FATHOMS®, modified GP2, or 

equivalent) prepared with MILLIQ® or equivalent deionized water 

(see Section 7, Dilution Water); or 

receiving water (available options)

Delete "FORTY 

FATHOMS®" from 

sentence.

...or artificial sea salts (HW MARINEMIX®, modified 

GP2, or equivalent)....

FORTY FATHOMS®, a 

commercial product, is no 

longer available. Equivalent 

commercial products, such as 

HW MARINEMIX®, are 

acceptable replacements.

WET Methods Errata Sheet 7 October 2014
Errata for Methods for Measuring the Acute Toxicity of Effluents and Receiving Waters to Freshwater and Marine Organisms, Fifth Edition. U.S. Environmental 

Protection Agency, Office of Water, Washington, DC EPA-821-R-02-012, October 2002

Text Location Current Text Correction Being Made Corrected Text Reason For Correction 

Section 11, Acute 

Toxicity Data Analysis, 

Section 11.2.4.3, Subbullet 6, The TrimmedKarber Method, Page 79

A computer program which

estimates the LC50 and associated

95% confidence interval using the 

Trimmed-Karber Method, can be

obtained through the 

Environmental Monitoring and

Support Laboratory (EMSL), 26

W. Martin Luther King Drive,

Cincinnati, OH 45268. The 

program can be obtained from 

EMSL-Cincinnati by sending a 

diskette with a written request to

the above address.

Replace "...through the 

Environmental Monitoring 

and Support...to the above 

address." with 

replacement software 

information.

...can be obtained by clicking on "contact us" at 

http://www.epa.gov/med. Any software capable of 

calculating the LC50 and associated 95% confidence 

interval using the Trimmed-Karber method can be used. 

Computer program is no 

longer available from 

Cincinnati. It can be 

obtained by sending a request 

to EPA using the link. Any 

software capable of 

completing the calculations is 

acceptable.

Section 11, Acute 

Toxicity Data Analysis, 

Section 11.2.5.3, Subbullet 2, The Probit 

Method, Page 81

A machine-readable, compiled,

version of a computer program to

estimate the LC1 and LC50 and 

associated 95% confidence

intervals using the Probit Method

can be obtained from EMSLCincinnati by sending a diskette 

with a written request to the 

Environmental Monitoring 

Systems Laboratory, 26 W. Martin

Luther King Drive, Cincinnati,

OH 45268.

Replace "...from EMSLCincinnati...Cincinnati, 

OH 45268" with 

replacement software 

information.

...can be obtained by clicking on "contact us" at 

http://www.epa.gov/med. Any software capable of 

calculating the LC1 and LC50 and associated 95% 

confidence interval using the Probit method can be used.

Computer program is no 

longer available from EMSLCincinnati. It can be obtained 

by sending a request to EPA 

using the link. Any software 

capable of completing the 

calculations is acceptable.

Appendix A, 

Distribution, Life Cycle, 

Taxonomy, and Culture 

Methods, Section 3.12.1, 

Culture Water, Page 218

Artificial seawater is prepared by 

dissolving FORTY-FATHOMS®

or equivalent artificial sea salts in 

deionized water to a salinity of 20-

30‰. 

Replace "FORTYFATHOMS®" with "HW 

MARINEMIX®" in 

sentence.

...prepared by dissolving HW MARINEMIX® or 

equivalent artificial sea salts....

FORTY FATHOMS®, a 

commercial product, is no 

longer available. Equivalent 

commercial products, such as 

HW MARINEMIX®, are 

acceptable replacements.

WET Methods Errata Sheet 8 October 2014
Errata for Short-term Methods for Estimating the Chronic Toxicity of Effluents and Receiving Waters to Freshwater Organisms, Fourth Edition. U.S. Environmental 

Protection Agency, Office of Water, Washington, DC EPA/821/R–02/013, October 2002

Text Location Current Text Correction Being Made Corrected Text Reason For Correction 

Section 4, Quality 

Assurance, Section 

4.4.1, Laboratory Water 

Used for Culturing and 

Test Dilution Water, 

Page 10 

Water used for culturing and test 

dilution should be analyzed for 

toxic metals and organics at least 

annually or whenever difficulty is

encountered in meeting minimum 

acceptability criteria for control 

survival and reproduction or 

growth.

Add "test" between 

"minimum" and 

"acceptability criteria." 

Water used for culturing and test dilution should be 

analyzed for toxic metals and organics at least annually 

or whenever difficulty is encountered in meeting 

minimum test acceptability criteria for control survival 

and reproduction or growth.

Clarification.

Section 4, Quality 

Assurance, Section 

4.8.3, Food Quality, 

Page 11

New batches of food used in

culturing and testing should be 

analyzed for toxic organics and 

metals or whenever difficulty is 

encountered in meeting minimum 

acceptability criteria for control 

survival and reproduction or 

growth.

Add "test" between 

"minimum" and

"acceptability criteria." 

New batches of food used in culturing and testing should 

be analyzed for toxic organics and metals or whenever 

difficulty is encountered in meeting minimum test 

acceptability criteria for control survival and 

reproduction or growth.

Consistency correction with 

Section 4.8.3 in "Acute 

Freshwater and Marine 

Manual." 

Section 4, Quality 

Assurance, Section 

4.8.3, Food Quality, 

Page 11

...is encountered in meeting 

minimum acceptability criteria for

control survival and reproduction 

or growth. 

Add "test" between 

"minimum" and 

"acceptability criteria" in 

first sentence.

...is encountered in meeting minimum test acceptability 

criteria for control survival and reproduction or growth. 

Consistency correction with 

Section 4.8.3 in "Acute 

Freshwater and Marine 

Manual" and Section 4.8.3 in 

"Chronic Marine and 

Estuarine Manual."

Section 4, Quality 

Assurance Section 4.16, 

Documenting Ongoing 

Laboratory Performance, 

Page 17

Replace the graphs in 

Figure 1 with log scale 

graphs. Add explanatory 

text to Figure 1 legend. 

Correction and clarification. 

Cusum charting can either 

plot anti-logarithm values for 

historical mean and warning 

limits on the graph with the 

y-axis scaled as logarithm or 

plot logarithm values for 

historical mean and warning 

limits on the graph with the 

y-axis as normal arithmetic 

scaling. The figures A and B 

are revised to match the text 

in the manual and are shown 

using a log format. Cusum 

charts may be plotted using 

antilogarithm values so that 

the original units (e.g., g/L) 

are displayed on the graph.

The log values on a log 

scaled y-axis or anti-log

WET Methods Errata Sheet 9 October 2014
Errata for Short-term Methods for Estimating the Chronic Toxicity of Effluents and Receiving Waters to Freshwater Organisms, Fourth Edition. U.S. Environmental 

Protection Agency, Office of Water, Washington, DC EPA/821/R–02/013, October 2002

Text Location Current Text Correction Being Made Corrected Text Reason For Correction 

Figure 1. Control (cusum) charts: For NOEC results 

(hypothesis estimates) (A): plot the value of the NOEC 

and the values of the adjacent test concentrations of 

NOEC as the upper and lower limits. For LC, EC or IC 

results (point estimates) (B) plot the actual point 

estimate, the cumulative mean (central tendency, middle 

line) and the upper and lower control limits (± 2S). The 

upper and lower control limits are calculated as the 

average IC, EC, LC value (± 2S) and plotted on the 

chart as horizontal lines serving as visual indicators of 

any trend of successive effect endpoints that are 

divergent results. The cusum chart should plot logarithm 

of concentration on the vertical axis against the date of 

the test or test number on the horizontal axis. Successive 

toxicity endpoints (i.e., IC25) should be plotted and 

examined to determine if the results are within 

prescribed limits. Cusum charts can either plot the data 

with the x-axis scaled as logarithms as normal arithmetic 

scaling or plot the anti-logarithm values on the graph 

with the y-axis scaled as logarithm. The log values on a 

log scaled y-axis or antilog values on an arithmetic scale 

y-axis are not be mixed on a chart.

values on an arithmetic scale 

y-axis are not be mixed on a 

chart. Many graphics 

programs do not allow 

enlarging the log scale axis 

sufficiently (i.e., expanding 

within a log unit), and trends 

can be difficult to view; 

therefore graphs using 

antilogarithm values may be

preferable. Individual LC, 

EC or IC data should be 

plotted on a graph. The graph 

should present the mean of 

the logarithmic LC, EC or 

ICs for the central tendency. 

The upper and lower control 

limits are calculated as the 

average LC, EC or IC value 

plus-and-minus two standard 

deviations and plotted on the 

chart as horizontal lines, 

serving as visual indicators of 

any trend of successive effect 

endpoints that are divergent 

results. For NOEC toxicity 

values, the control limits 

would consist of one 

concentration interval above 

and below the concentration 

representing the central 

tendency. NOEC values 

cannot be averaged (Section 

4.14.5); actual values and 

adjacent upper and lower 

concentrations to the NOEC 

value are plotted using the 

logarithm procedures 

described in Section 4.14.5 

and Section 9, Chronic 

Toxicity Test Endpoints and 

Data Analysis) the NOECLOEC interval is used for the 

WET Methods Errata Sheet 10 October 2014
Errata for Short-term Methods for Estimating the Chronic Toxicity of Effluents and Receiving Waters to Freshwater Organisms, Fourth Edition. U.S. Environmental 

Protection Agency, Office of Water, Washington, DC EPA/821/R–02/013, October 2002

Text Location Current Text Correction Being Made Corrected Text Reason For Correction 

cusum chart. Reference: 

Environment Canada. 2009. 

Environment Canada’s 

Guidance on Using 

Logarithms for Reference 

Toxicant Warning Chart. 

Biological Methods Section, 

September.

Section 11, Fathead 

Minnow, Pimephales 

promelas, Larval 

Survival and Growth 

Test Method, Section 

11.3.6.1.2, Interferences, 

Page 57

Total ammonia concentrations

greater than 5 mg/L in the 100%

effluent are an indicator that

toxicity observed in the test may

be due to ammonia (USEPA,

1992).

Replace "the 100% 

effluent are" with "in any 

concentration tested may 

be...." 

Total ammonia concentrations greater than 5 mg/L in any 

concentration tested may be an indicator that toxicity

observed in the test may be due to ammonia (USEPA,

1992).

Clarification.

Section 11, Fathead 

Minnow, Pimephales 

promelas, Larval 

Survival and Growth 

Test Method 1000.0, 

Section 11.10.1.2.3, Test 

Procedures, Page 65

Sufficient test solution 

(approximately 1500 mL) is 

prepared at each effluent 

concentration to provide 400 mL 

additional volume for chemical 

analyses at the high, medium, and 

low test concentrations.

Replace "at the high, 

medium and low test" with 

"for all" test 

concentrations.

Sufficient test solution (approximately 1500 mL) is 

prepared at each effluent concentration to provide 400 

mL additional volume for chemical analysis for all test 

concentrations.

Clarification. All solutions 

need to have sufficient 

volume prepared, not just the 

high, medium, and low test 

concentrations.

Section 11, Fathead 

Minnow, Pimephales 

promelas, Larval 

Survival and Growth 

Test Method 1000.0, 

Section 11.10.6.1.2, 

Observations During the 

Test, Page 67

Temperature and pH are measured 

at the end of each 24-h exposure 

period.... 

Insert "beginning and"

between "measured at the" 

and "end of each." 

Temperature and pH are measured at the beginning and 

end of each 24-h exposure period.... 

Clarification.

Section 11, Fathead 

Minnow, Pimephales 

promelas, Larval 

Survival and Growth 

Test Method 1000.0, 

Section 11.10.6.1.4, 

Observations During the 

Test, Page 67

Conductivity, alkalinity, and 

hardness are measured in each 

new sample (100% effluent or 

receiving water) and in the 

control.

Inset "at the beginning of 

the test for all test 

concentrations" between 

"hardness are measured" 

and "in each new sample." 

Delete "(100% effluent or 

receiving water)" from 

sentence. Insert "before 

they are dispensed to the 

test chambers" to the end 

of the sentence.

Conductivity, alkalinity, and hardness are measured at 

the beginning of the test for all test concentrations in 

each new sample and in the control before they are 

dispensed to the test chambers.

Clarification. Conductivity, 

alkalinity and hardness are 

measured on all test 

concentrations with new 

samples, not only the 100% 

effluent or receiving waters. 

WET Methods Errata Sheet 11 October 2014
Errata for Short-term Methods for Estimating the Chronic Toxicity of Effluents and Receiving Waters to Freshwater Organisms, Fourth Edition. U.S. Environmental 

Protection Agency, Office of Water, Washington, DC EPA/821/R–02/013, October 2002

Text Location Current Text Correction Being Made Corrected Text Reason For Correction 

Section 12, Fathead 

Minnow, Pimephales 

promelas, EmbryoLarval Survival and 

Teratogenicity Test 

Method 1001.0, Section 

12.3.5.1.2, Interferences, 

Page 113

Total ammonia concentrations

greater than 5 mg/L in the 100%

effluent are an indicator that

toxicity observed in the test may

be due to ammonia (USEPA,

1992). 

Replace "the 100% 

effluent are" with "in any 

concentration tested may 

be".... 

Total ammonia concentrations greater than 5 mg/L in any 

concentration tested may be an indicator that toxicity

observed in the test may be due to ammonia (USEPA,

1992).

Clarification.

Section 12, Fathead 

Minnow, Pimephales 

promelas, EmbryoLarval Survival and 

Teratogenicity Test 

Method 1001.0, Section 

12.10.1.2.3, Test 

Solutions, Page 117

Sufficient test solution 

(approximately 1000 mL) is 

prepared at each effluent

concentration to provide 400 mL 

additional volume for chemical 

analyses.

Change "1000 mL" to 

"1500 mL." Add "for all 

test concentrations" to end 

of sentence.

Sufficient test solution (approximately 1500 mL) is 

prepared at each effluent concentration to provide 400 

mL additional volume for chemical analyses for all test 

concentrations.

Clarification. A volume of 

1000 mL is not sufficient for 

the test. Consistency 

correction with Section 

11.10.1.2.3 in "Chronic 

Freshwater Manual." All 

solutions need to have 

sufficient volume prepared, 

not just the high, medium and 

low test concentrations.

Section 12, Fathead 

Minnow, Pimephales 

promelas, EmbryoLarval Survival and 

Teratogenicity Test 

Method 1001.0, Section 

12.10.6.1.2, 

Observations During the 

Test, Page 119

Temperature and pH are measured 

at the end of each 24-h exposure 

period in at least one test 

chamber.... 

Add "beginning and"

between "the" and "end." 

Temperature and pH are measured at the beginning and 

end of each 24-h exposure period in at least one test 

chamber.... 

Clarification. 

Section 12, Fathead 

Minnow, Pimephales 

promelas, EmbryoLarval Survival and 

Teratogenicity Test 

Method 1001.0, Section 

12.10.6.1.4, 

Observations During the 

Test, Page 119

Conductivity, alkalinity, and 

hardness are measured in each 

new sample (100% effluent or 

receiving water) and in the 

control.

Replace "in each new 

sample (100% effluent or 

receiving water) and in 

the" with "at the beginning 

of the test for all test 

concentrations for each 

new sample and control in 

the before they are 

dispensed to the test 

chambers."

Conductivity, alkalinity, and hardness are measured at 

the beginning of the test for all test concentrations for 

each new sample and in the control before they are 

dispensed to the test chambers.

Clarification. Conductivity, 

alkalinity and hardness are 

measured on all test 

concentrations with new 

samples, not only the 100% 

effluent or receiving waters. 

WET Methods Errata Sheet 12 October 2014
Errata for Short-term Methods for Estimating the Chronic Toxicity of Effluents and Receiving Waters to Freshwater Organisms, Fourth Edition. U.S. Environmental 

Protection Agency, Office of Water, Washington, DC EPA/821/R–02/013, October 2002

Text Location Current Text Correction Being Made Corrected Text Reason For Correction 

Section 13, Daphnid, 

Ceriodaphnia dubia,

Survival and 

Reproduction Test 

Method 1002.0, Section 

13.3.6.1.2 Interferences, 

Page 142

Total ammonia concentrations

greater than 5 mg/L in the 100%

effluent are an indicator that

toxicity observed in the test may

be due to ammonia (USEPA,

1992).

Replace "the 100% 

effluent are" with "in any 

concentration tested may 

be." 

Total ammonia concentrations greater than 5 mg/L in any 

concentration tested may be an indicator that toxicity

observed in the test may be due to ammonia (USEPA,

1992).

Clarification.

Section 13, Daphnid,

Ceriodaphnia dubia, 

Survival and 

Reproduction Test 

Method 1002.0, Section 

13.10.6.1.2, 

Observations During the 

Test, Page 157

Temperature and pH are measured 

at the end of each 24-h exposure 

period in at least one test 

chamber....

Add "beginning and"

between "at the" and "of 

each 24-h." 

Temperature and pH are measured at the beginning and 

end of each 24-h exposure period in at least one test 

chamber.... 

Clarification

Section 13, Daphnid, 

Ceriodaphnia dubia,

Survival and 

Reproduction Test 

Method 1002.0, Section 

13.10.6.1.4, 

Observations During the 

Test, Page 157

Conductivity, alkalinity, and 

hardness are measured in each 

new sample (100% effluent or 

receiving water) and in the 

control.

Add "at the beginning of 

the test for all test 

concentrations" between 

"are measured" and "in 

each." Delete "(100% 

effluent or receiving 

water)." Add "before they 

are dispersed to the test 

chambers" to end of the 

sentence.

Conductivity, alkalinity, and hardness are measured at 

the beginning of the test for all test concentrations in 

each new sample and in the control before they are 

dispensed to the test chambers.

Clarification.

WET Methods Errata Sheet 13 October 2014
Errata for Short-term Methods for Estimating the Chronic Toxicity of Effluents and Receiving Waters to Freshwater Organisms, Fourth Edition. U.S. Environmental 

Protection Agency, Office of Water, Washington, DC EPA/821/R–02/013, October 2002

Text Location Current Text Correction Being Made Corrected Text Reason For Correction 

Section 13, Daphnid, 

Ceriodaphnia dubia,

Survival and 

Reproduction Test 

Method 1002.0, Section 

13.13.3.1, Example of 

Analysis of the Daphnid 

Reproduction Data, Page 

170

The response used in the statistical 

analysis is the number of young 

produced per adult female, which 

is determined by taking the total 

number of young produced until 

either the time of death of the 

adult or the end of the experiment, 

whichever comes first. In this 

three-brood test, offspring from 

fourth or higher broods should not 

be counted and should not be 

included in the total number of 

neonates produced during the test. 

An animal that dies before 

producing young, if it has not been 

identified as a male, would be 

included in the analysis with zero 

entered as the number of young 

produced. The subsequent 

calculation of the mean number of 

live young produced per adult 

female for each toxicant provides 

a combined measure of the 

toxicant's effect on both mortality 

and reproduction.

Add word "live" in the 

first (twice), second 

(once), and third (twice) 

sentences. 

The response used in the statistical analysis is the 

number of live young produced per adult female, which 

is determined by taking the total number of live young 

produced until either the time of death of the adult or the 

end of the experiment, whichever comes first. In this 

three-brood test, offspring from fourth or higher broods 

should not be counted and should not be included in the 

total number of live neonates produced during the test. 

An animal that dies before producing live young, if it has 

not been identified as a male, would be included in the 

analysis with zero entered as the number of live young 

produced. The subsequent calculation of the mean 

number of live young produced per adult female for each 

toxicant provides a combined measure of the toxicant's 

effect on both mortality and reproduction.

Clarification.

Section 14, Green Alga, 

Selenastrum 

capricornutum, Growth 

Test Method 1003.0, 

Section 14.10.1.2.3, Test 

Procedures, Page 204

Sufficient test solution 

(approximately 900 or 1500 mL) 

is prepared at each effluent 

concentration to provide 400 mL 

additional volume for chemical 

analyses at the high, medium, and 

low test concentrations.

Replace "at the high, 

medium, and low" with 

"for all test." 

Sufficient test solution (approximately 900 or 1500 mL) 

is prepared at each effluent concentration to provide 400 

mL additional volume for chemical analyses for all test 

concentrations.

Clarification. All solutions 

need to have sufficient 

volume prepared, not just the 

high, medium, and low test 

concentrations.

Section 14, Green Alga, 

Selenastrum 

capricornutum, Growth 

Test Method 1003.0, 

Section 14.10.2.3.1, Start 

of Test, Page 206

Alkalinity, hardness, and 

conductivity are measured at the 

beginning of the test in the high, 

medium, and low effluent 

concentrations and control before 

they are dispensed to the test 

chambers and the data recorded on 

the data sheet (Figure 1).

Replace "in the high, 

medium, and low" with 

"for all test." Add/change

words "in each new 

sample and in the" 

between "concentrations" 

and "control." 

Alkalinity, hardness, and conductivity are measured at 

the beginning of the test for all test concentrations in 

each new sample and in the control before they are 

dispensed to the test chambers and the data recorded on 

the data sheet (Figure 1). 

Clarification. 

WET Methods Errata Sheet 14 October 2014
Errata for Short-term Methods for Estimating the Chronic Toxicity of Effluents and Receiving Waters to Freshwater Organisms, Fourth Edition. U.S. Environmental 

Protection Agency, Office of Water, Washington, DC EPA/821/R–02/013, October 2002

Text Location Current Text Correction Being Made Corrected Text Reason For Correction 

Section 14, Green Alga, 

Selenastrum 

capricornutum, Growth 

Test Method 1003.0, 

Section 14.10.5.1.2, 

Observations During the 

Test, Page 206

Temperature and pH are measured 

at the end of each 24-h exposure 

period in at least one test flask....

Add "beginning and"

before "end." 

Temperature and pH are measured at the beginning and 

end of each 24-h exposure period in at least one test 

flask....

Clarification. The algae test 

does not have a daily renewal 

of test solution.

Appendix C, Dunnett's 

Procedure, Section 2.5, 

Computer Calculations, 

Page 275

...Cincinnati, OH 45268. A 

compiled executable version of 

the program can be obtained from 

EMSL-Cincinnati by sending a 

written request to EMSL at 3411 

Church Street, Cincinnati, OH 

45244. 

Replace "...EMSLCincinnati...." with 

replacement software 

information.

...can be obtained by clicking on "contact us" at 

http://www.epa.gov/med. Any software capable of 

calculating an analysis of variance (ANOVA) and a 

multiple comparison of treatment means with the control 

mean (Dunnett's Procedure) can be used.

Computer program is no 

longer available from EMSLCincinnati. It can be obtained 

by sending a request to EPA 

using the link. Any software 

capable of completing the 

calculations is acceptable.

Appendix I, Probit 

Analysis, 2, Page 309

A compiled, executable version of

the program can be obtained from

EMSL-Cincinnati by sending a 

written request to EMSL at 3411

Church Street, Cincinnati, OH 

45244.

Replace "...EMSLCincinnati...." with 

replacement software 

information.

...can be obtained by clicking on "contact us" at 

http://www.epa.gov/med. Any software capable of 

calculating the EC1 and EC 50 (or LC1 and LC50) and 

the associated 95% confidence interval using the Probit 

method can be used.

Computer program is no 

longer available from EMSLCincinnati. It can be obtained 

by sending a request to EPA 

using the link. Any software 

capable of completing the 

calculations is acceptable.

Appendix K, Trimmed 

Spearman-Karber 

Method, Section 10, 

Page 318

A computer program which

estimates the LC50 and associated

95% confidence interval using the 

Trimmed Spearman-Karber

Method, can be obtained from

EMSL-Cincinnati by sending a 

written request to EMSL, 3411 

Church Street, Cincinnati, OH 

45244.

Replace "...EMSLCincinnati...." with 

replacement software 

information.

...can be obtained by clicking on "contact us" at 

http://www.epa.gov/med. Any software capable of 

calculating the LC50 and associated 95% confidence 

interval using the Trimmed Spearman-Karber Method 

can be used.

Computer program is no 

longer available from EMSLCincinnati. It can be obtained 

by sending a request to EPA 

using the link. Any software 

capable of completing the 

calculations is acceptable.

Appendix M, Linear 

Interpolation Method, 

Section 7.1, Page 329

To obtain the program and

supporting documentation, send a

written request to EMSLCincinnati at 3411 Church Street,

Cincinnati, OH 45244.

Replace "...EMSLCincinnati...." with 

replacement software 

information.

...can be obtained by clicking on "contact us" at 

http://www.epa.gov/med. Any software capable of 

calculating the mean and standard deviation of the 

bootstrapped ICp and associated 95% confidence 

intervals can be used.

Computer program is no 

longer available from EMSLCincinnati. It can be obtained 

by sending a request to EPA 

using the link. Any software 

capable of completing the 

calculations is acceptable.

WET Methods Errata Sheet 15 October 2014
Errata for Methods for Measuring the Chronic Toxicity of Effluents and Receiving Waters to Marine and Estuarine Organisms, Third Edition. U.S. Environmental 

Protection Agency, Office of Water, Washington DC EPA/821/R–02/014, October 2002

Text Location Current Text Correction Being Made Corrected Text Reason For Correction 

Section 4, Quality 

Assurance, Section 

4.4.1, Laboratory Water 

Used For Culturing and 

Test Dilution Water, 

Page 10

The dilution water used in the 

toxicity tests may be natural 

seawater, hypersaline brine 

(100‰) prepared from natural 

seawater, or artificial seawater 

prepared from commercial sea 

salts, such as FORTY FATHOMS®

or HW MARINEMIX®, if 

recommended in the method. 

Delete "FORTY 

FATHOMS® or," from 

sentence. Add "or 

equivalent" after 

"MARINEMIX®" in 

sentence.

....or artificial seawater prepared from commercial sea 

salts, such as HW MARINEMIX® or equivalent if 

recommended in the method. 

FORTY FATHOMS®, a 

commercial product, is no 

longer available. Equivalent 

commercial products, such as 

HW MARINEMIX®, are 

acceptable replacements.

Section 4, Quality 

Assurance, Section 

4.8.3, Food Quality, 

Page 11

...the food should not be used (for 

analytical methods, see AOAC, 

1990; and USDA, 1989). 

Add sentence "For foods 

(e.g., flake food) which 

are used to culture and test 

organisms, the quality of 

the food should meet the 

requirements for 

laboratory water used for 

culturing and test dilution 

water as described in 

Section 4.4 above at the 

concentration in the 

culture or test solutions." 

after end of sentence. 

...the food should not be used (for analytical methods, 

see AOAC, 1990; and USDA, 1989). For foods (e.g., 

flake food) which are used to culture and test organisms, 

the quality of the food should meet the requirements for 

laboratory water used for culturing and test dilution 

water as described in Section 4.4 above at the 

concentration in the culture or test solutions. 

Analysis should be at the 

concentration in the culture 

or test waters; consistency 

correction with Section 4.8.3 

"Acute Freshwater and 

Marine Manual" and Section 

4.8.3 "Chronic Freshwater 

Manual"

Section 4, Quality 

Assurance, Section 

4.8.4, Food Quality, 

Page 11 

For foods (e.g., YCT) which are 

used to culture....

Replace "YCT" with 

"flake food."

For foods (e.g., flake food) which are used to culture.... Correction. The food YCT is 

not used in any of the marine 

methods.

WET Methods Errata Sheet 16 October 2014
Errata for Methods for Measuring the Chronic Toxicity of Effluents and Receiving Waters to Marine and Estuarine Organisms, Third Edition. U.S. Environmental 

Protection Agency, Office of Water, Washington DC EPA/821/R–02/014, October 2002

Text Location Current Text Correction Being Made Corrected Text Reason For Correction 

Section 4, Quality 

Assurance, Section 4.16, 

Documenting Ongoing 

Laboratory Performance, 

Page 11

Replace the graphs in 

Figure 1 with log scale 

graphs. Add explanatory 

text to Figure 1 legend.

Figure 1. Control (cusum) charts: For NOEC results 

(hypothesis estimates) (A): plot the value of the NOEC 

and the values of the adjacent test concentrations of 

NOEC as the upper and lower limits. For LC, EC or IC 

results (point estimates) (B) plot the actual point 

estimate, the cumulative mean (central tendency, middle 

line) and the upper and lower control limits (± 2S). The 

upper and lower control limits are calculated as the 

average IC, EC, LC value (± 2S) and plotted on the 

chart as horizontal lines serving as visual indicators of 

any trend of successive effect endpoints that are 

divergent results. The cusum chart should plot logarithm 

of concentration on the vertical axis against the date of 

the test or test number on the horizontal axis. Successive 

toxicity endpoints (i.e., IC25) should be plotted and 

examined to determine if the results are within 

prescribed limits. Cusum charts can either plot the data 

with the x-axis scaled as logarithms as normal arithmetic

Correction and clarification. 

Cusum charting can either 

plot anti-logarithm values for 

historical mean and warning 

limits on the graph with the 

y-axis scaled as logarithm or 

plot logarithm values for 

historical mean and warning 

limits on the graph with the 

y-axis as normal arithmetic 

scaling. The figures A and B 

are revised to match the text 

in the manual and are shown 

using a log format. Cusum 

charts may be plotted using 

antilogarithm values so that 

the original units (e.g., g/L) 

are displayed on the graph. 

The log values on a log 

scaled y-axis or antilog 

values on an arithmetic scale 

y-axis are not be mixed on a 

chart. Many graphics 

programs do not allow 

enlarging the log scale axis 

sufficiently (i.e., expanding 

within a log unit), and trends 

can be difficult to view; 

therefore graphs using 

antilogarithm values may be 

preferable. Individual LC, 

EC or IC data should be 

plotted on a graph. The graph 

should present the mean of 

the logarithmic LC, EC or 

ICs for the central tendency. 

The upper and lower control 

limits are calculated as the 

average LC, EC or IC value 

plus-and-minus two standard 

deviations and plotted on the 

chart as horizontal lines,

serving as visual indicators of

WET Methods Errata Sheet 17 October 2014
Errata for Methods for Measuring the Chronic Toxicity of Effluents and Receiving Waters to Marine and Estuarine Organisms, Third Edition. U.S. Environmental 

Protection Agency, Office of Water, Washington DC EPA/821/R–02/014, October 2002

Text Location Current Text Correction Being Made Corrected Text Reason For Correction 

scaling or plot the anti-logarithm values on the graph 

with the y-axis scaled as logarithm. The log values on a 

log scaled y-axis or antilog values on an arithmetic scale 

y-axis are not be mixed on a chart.

any trend of successive effect 

endpoints that are divergent 

results. For NOEC toxicity 

values, the control limits 

would consist of one 

concentration interval above 

and below the concentration 

representing the central 

tendency. NOEC values 

cannot be averaged (Section 

4.14.5); actual values and 

adjacent upper and lower 

concentrations to the NOEC 

value are plotted using the 

logarithm procedures 

described in Section 4.14.5 

and Section 9, Chronic 

Toxicity Test Endpoints and 

Data Analysis) the NOECLOEC interval is used for the 

cusum chart. Reference: 

Environment Canada. 2009.

Environment Canada’s 

Guidance on Using 

Logarithms for Reference 

Toxicant Warning Chart.

Biological Methods Section, 

September.

Section 4, Quality 

Assurance, Section 

4.17.1, Reference 

Toxicants, Page 16

. . .requiring aquatic toxicity tests. 

EMSL-Cincinnati hopes to release 

EPA-certified solutions of 

cadmium and copper for use as 

reference toxicants through 

cooperative research and 

development agreements with 

commercial suppliers, and will 

continue to develop additional 

reference toxicants for future 

release. Standard reference 

materials....

Delete sentence "EMSLCincinnati hopes to ...". . .requiring aquatic toxicity tests. Standard reference 

materials....

EMSL-Cincinnati no longer 

provides reference toxicants. 

WET Methods Errata Sheet 18 October 2014
Errata for Methods for Measuring the Chronic Toxicity of Effluents and Receiving Waters to Marine and Estuarine Organisms, Third Edition. U.S. Environmental 

Protection Agency, Office of Water, Washington DC EPA/821/R–02/014, October 2002

Text Location Current Text Correction Being Made Corrected Text Reason For Correction 

Section 5, Facilities, 

Equipment, and 

Supplies, Section 5.1, 

General Requirements, 

Page 19

Water used for rearing, holding, 

acclimating, and testing organisms 

may be natural seawater or water 

made up from hypersaline brine 

derived from natural seawater, or

water made up from reagent grade 

chemicals (GP2) or commercial 

(FORTY FATHOMS® or HW 

MARINEMIX®) artificial sea salts 

when specifically recommended in 

the method.

Delete "FORTY 

FATHOMS®" from 

sentence.

...or water made up from reagent grade chemicals (GP2) 

or commercial (HW MARINEMIX®) artificial sea 

salts....

FORTY FATHOMS®, a 

commercial product, is no 

longer available. 

Section 6, Test 

Organisms, Section 

6.6.4, Transportation to 

the Test Site, Page 25

...by adding sufficient dry ocean 

salts, such as FORTY 

FATHOMS®, or equivalent, GP2, 

or hypersaline brine.

Delete "FORTY 

FATHOMS®, or 

equivalent," in sentence.

...by adding sufficient dry ocean salts, such as GP2, or 

hypersaline brine.

FORTY FATHOMS®, a 

commercial product, is no 

longer available. 

Section 7, Dilution 

Water, Section 7.2.1, 

Types of Dilution, Page 

26

Standard, synthetic, dilution water 

is prepared with deionized water 

and reagent grade chemicals (GP2) 

or commercial sea salts (FORTY 

FATHOMS®, HW 

MARINEMIX®) (Table 3). 

Delete "FORTY 

FATHOMS®," and add "or 

equivalent" after "HW 

MARINEMIX®" in 

sentence.

...or commercial artificial sea salts (HW MARINE 

MIX® or equivalent) (Table 3).

FORTY FATHOMS®, a 

commercial product, is no 

longer available. Equivalent 

commercial products, such as 

HW MARINEMIX®, are 

acceptable replacements.

Section 7, Dilution 

Water, Section 7.2.3.2, 

Standard Synthetic Sea 

Water, Page 27

Synthetic seawater can also be 

prepared by adding commercial sea 

salts, such as FORTY 

FATHOMS®, HW 

MARINEMIX®, or equivalent, to 

deionized water. For example, 

thirty-one parts per thousand 

(31‰) FORTY FATHOMS® can 

be prepared by dissolving 31 g of 

sea salts per liter of deionized 

water. The source water for the 

deionizer can be ground water or 

tap water.

Delete "FORTY 

FATHOMS®" from first 

sentence. Delete second 

sentence.

Synthetic seawater can be prepared by adding 

commercial sea salts, such as HW MARINEMIX®, or 

equivalent, to deionized water. The source water for the 

deionizer....

FORTY FATHOMS®, a 

commercial product, is no 

longer available. Equivalent 

commercial products, such as 

HW MARINEMIX®, are 

acceptable replacements.

WET Methods Errata Sheet 19 October 2014
Errata for Methods for Measuring the Chronic Toxicity of Effluents and Receiving Waters to Marine and Estuarine Organisms, Third Edition. U.S. Environmental 

Protection Agency, Office of Water, Washington DC EPA/821/R–02/014, October 2002

Text Location Current Text Correction Being Made Corrected Text Reason For Correction 

Section 7, Dilution 

Water, Section 7.3.3, 

Use of Receiving Water 

as Dilution Water, Page 

28

Where necessary, the salinity of a 

surface water can be increased by 

the addition of artificial sea salts, 

such as FORTY FATHOMS®, HW 

MARINEMIX®, or equivalent, 

GP2, a natural seawater of higher 

salinity, or hypersaline brine. 

Delete "FORTY 

FATHOMS®," from 

sentence.

... addition of artificial sea salts, such as HW 

MARINEMIX®, or equivalent, GP2, a natural seawater....

FORTY FATHOMS®, a 

commercial product, is no 

longer available. Equivalent 

commercial products, such as 

HW MARINEMIX®, are 

acceptable replacements.

Section 11, Sheepshead 

Minnow, Cyprinodon 

variegatus, Larval 

Survival and Growth 

Test Method 1004.0, 

Section 11.3.6.1.2, 

Interferences, Page 56

Total ammonia concentrations

greater than 5 mg/L in the 100%

effluent are an indicator that

toxicity observed in the test may

be due to ammonia (USEPA,

1992).

Replace "the 100% 

effluent are" with "any 

concentration tested may 

be."

Total ammonia concentrations greater than 5 mg/L in any 

concentration tested may be an indicator that toxicity

observed in the test may be due to ammonia (USEPA,

1992).

Clarification.

Section 11, Sheepshead 

Minnow, Cyprinodon 

variegatus, Larval 

Survival and Growth 

Test Method 1004.0, 

Section 11.10.1.3.1, 

Dilution Water, Page 68

Dilution water may be 

uncontaminated natural seawater 

(receiving water), HSB prepared 

from natural seawater, or artificial

seawater prepared from FORTY 

FATHOMS® or GP2 sea salts (see 

Table 1 and Section 7, Dilution 

Water). 

Replace "FORTY 

FATHOMS®" with "HW 

MARINEMIX®, or 

equivalent," and replace 

"1" with "3" in sentence.

...or artificial seawater prepared from HW 

MARINEMIX®, or equivalent, or GP2 sea salts (see 

Table 3, Section 7....

FORTY FATHOMS®, a 

commercial product, is no 

longer available. Equivalent 

commercial products, such as 

HW MARINEMIX®, are 

acceptable replacements.

Consistency with Section 11, 

Table 3, Summary of Test 

Conditions..., Line 18, 

Dilution Water, Page 79

Section 11, Sheepshead 

Minnow, Cyprinodon 

variegatus, Larval 

Survival and Growth 

Test Method 1004.0, 

Section 11.10.1.2.3, 

Effluents, Table 2, 

Footnote 1, Page 69

The salinity of the initial 6,800 

mL of 100% effluent is adjusted to 

20‰ by adding 136 g of dry 

artificial sea salts (FORTY 

FATHOMS7

).

Replace "136 g of dry 

artificial sea salts (FORTY 

FATHOMS7)" with "an 

appropriate amount of dry 

artificial sea salts (e.g., 

HW MARINEMIX®)" in 

sentence.

...by adding an appropriate amount of dry artificial sea 

salts (e.g., HW MARINEMIX®).

FORTY FATHOMS®, a 

commercial product, is no 

longer available. Equivalent 

commercial products, such as 

HW MARINEMIX®, are 

acceptable replacements.

Section 11, Sheepshead 

Minnow, Cyprinodon 

variegatus, Larval 

Survival and Growth 

Test Method 1004.0, 

Section 11.10.7.1.2, 

Observations During the 

Test, Page 71

Temperature, pH, and salinity are 

measured at the end of each 24-h 

exposure period in one test 

chamber at each test concentration 

and in the control.

Add "beginning and"

between "at the" and "end 

of." Add "at least one"

between "period in" and 

"test chamber."

Temperature, pH, and salinity are measured at the 

beginning and end of each 24-hr exposure period in at 

least one test chamber at each test concentration and in 

the control.

Clarification.

WET Methods Errata Sheet 20 October 2014
Errata for Methods for Measuring the Chronic Toxicity of Effluents and Receiving Waters to Marine and Estuarine Organisms, Third Edition. U.S. Environmental 

Protection Agency, Office of Water, Washington DC EPA/821/R–02/014, October 2002

Text Location Current Text Correction Being Made Corrected Text Reason For Correction 

Section 11, Sheepshead 

Minnow, Cyprinodon 

variegatus, Larval 

Survival and Growth 

Test Method 1004.0, 

Section 11.11, Summary 

of Test Conditions and 

Test Acceptability 

Criteria, Table 3, Line 

18, Page 79

Uncontaminated source of natural 

seawater; deionized water mixed 

with hypersaline brine or artificial 

sea salts (HW MARINEMIX®, 

FORTY FATHOMS7

, GP2 or 

equivalent) (available options)

Delete "FORTY 

FATHOMS7," from 

sentence.

...or artificial sea salts (HW MARINEMIX®, GP2 or 

equivalent)....

FORTY FATHOMS®, a 

commercial product, is no 

longer available. Equivalent 

commercial products, such as 

HW MARINEMIX®, are 

acceptable replacements.

Section 12, Sheepshead 

Minnow, Cyprinodon 

variegatus, Embryo

Larval Survival and 

Tertatogenicity Test 

Method 1005.0, Section

12.6.10.3.8, Reagents 

and Consumable 

Materials, Table 1, 

Footnote 1, Page 126

The salinity of the initial 4000 mL 

of 100% effluent is adjusted to 

20‰ by adding 80 g of dry 

artificial sea salts (HW 

MARINEMIX or FORTY 

FATHOMS®), and mixing for 1 h.

Delete "FORTY 

FATHOMS®," from 

sentence, add "®" after 

"MARINEMIX", and add 

"equivalent" after "or" in 

sentence.

...dry artificial sea salts (HW MARINEMIX® or 

equivalent) and mixing for 1 h.

FORTY FATHOMS®, a 

commercial product, is no 

longer available. Equivalent 

commercial products, such as 

HW MARINEMIX®, are 

acceptable replacements.

Section 12, Sheepshead 

Minnow, Cyprinodon 

variegatus, Embryo 

Larval Survival and 

Tertatogenicity Test 

Method 1005.0, Section

12.10.1.3.1, Dilution 

Water, Page 131

Dilution water may be 

uncontaminated natural seawater 

(receiving water), HSB prepared 

from natural seawater, or artificial 

seawater prepared from FORTY 

FATHOMS® or GP2 sea salts (see 

Table 3 in Section 7, Dilution 

Water). 

Replace "FORTY 

FATHOMS®" with "HW 

MARINEMIX® or 

equivalent" in sentence.

...artificial seawater prepared from HW MARINEMIX®

or equivalent or GP2 sea salts....

FORTY FATHOMS®, a 

commercial product, is no 

longer available. Equivalent 

commercial products, such as 

HW MARINEMIX®, are 

acceptable replacements.

Consistency with Section 12, 

Table 2, Summary of Test 

Conditions..., Line 16, 

Dilution Water, Page 137

Section 12, Sheepshead 

Minnow, Cyprinodon 

variegatus, Embryo 

Larval Survival and 

Tertatogenicity Test 

Method 1005.0, Section 

12.10.6.1.2, 

Observations During the 

Test, Page 132

Temperature, pH, and salinity are 

measured at the end of each 24-h 

exposure period in one test 

chamber at each test concentration 

and in the control.

Add "beginning and"

between "measured at the"

and "end of each." Add 

"at least" between 

"exposure period in" and 

"one test chamber."

Temperature, pH, and salinity are measured at the 

beginning and end of each 24-hr exposure period in at 

least one test chamber at each test concentration and in 

the control. 

Clarification.

WET Methods Errata Sheet 21 October 2014
Errata for Methods for Measuring the Chronic Toxicity of Effluents and Receiving Waters to Marine and Estuarine Organisms, Third Edition. U.S. Environmental 

Protection Agency, Office of Water, Washington DC EPA/821/R–02/014, October 2002

Text Location Current Text Correction Being Made Corrected Text Reason For Correction 

Section 12, Sheepshead 

Minnow, Cyprinodon 

variegatus, Embryo 

Larval Survival and 

Tertatogenicity Test 

Method 1005.0, Section

12.12.1, Summary of 

Test Conditions and Test 

Acceptability Criteria, 

Table 2, Line 16, 

Dilution Water, Page 

137

Uncontaminated source of natural 

seawater; deionized water mixed 

with hypersaline brine or artificial 

sea salts (HW MARINEMIX®, 

FORTY FATHOMS®, GP2, or 

equivalent) (available options)

Delete "FORTY 

FATHOMS®," from 

sentence.

...or artificial sea salts (HW MARINEMIX®, GP2, or 

equivalent)....

FORTY FATHOMS®, a 

commercial product, is no 

longer available. Equivalent 

commercial products, such as 

HW MARINEMIX®, are 

acceptable replacements.

Section 13, Inland

Silverside, Menidia 

beryllina, Larval

Survival And Growth 

Method 1006.0, Section

13.10.1.3.1, Dilution 

Water, Page 169

Dilution water may be 

uncontaminated natural seawater 

(receiving water), HSB prepared 

from natural seawater, or artificial 

seawater prepared from FORTY 

FATHOMS® or GP2 sea salts (see 

Table 3 in Section 7, Dilution 

Water). 

Replace "FORTY 

FATHOMS®" with "HW 

MARINEMIX®" in 

sentence.

...or artificial seawater prepared from HW 

MARINEMIX® or GP2....

FORTY FATHOMS®, a 

commercial product, is no 

longer available. Equivalent 

commercial products, such as 

HW MARINEMIX®, are 

acceptable replacements.

Consistency with Section 13, 

Table 3, Summary of Test 

Conditions..., Line 18, 

Dilution Water, Page 179.

Section 13, Inland

Silverside, Menidia 

beryllina, Larval

Survival And Growth 

Method 1006.0, Section 

13.10.7.1.2, 

Observations During the 

Test, Page 171

Temperature, pH, and salinity are 

measured at the end of each 24-hr 

exposure period in one test 

chamber at all test concentrations 

and in the control. 

Add "beginning and"

between "measured at the"

and "end of each." Add 

"at least" between 

"exposure period in" and 

"one test chamber."

Temperature, pH, and salinity are measured at the 

beginning and end of each 24-hr exposure period in at 

least one test chamber at all test concentrations and the 

control. 

Clarification.

WET Methods Errata Sheet 22 October 2014
Errata for Methods for Measuring the Chronic Toxicity of Effluents and Receiving Waters to Marine and Estuarine Organisms, Third Edition. U.S. Environmental 

Protection Agency, Office of Water, Washington DC EPA/821/R–02/014, October 2002

Text Location Current Text Correction Being Made Corrected Text Reason For Correction 

Section 13, Inland

Silverside, Menidia 

beryllina, Larval

Survival And Growth 

Method 1006.0, Section

13.11, Summary of Test 

Conditions and Test 

Acceptability Criteria, 

Table 3, Line 18, 

Dilution Water, Page 

179

Uncontaminated source of natural 

sea water, artificial seawater; 

deionized water mixed with 

hypersaline brine or artificial sea 

salts (HW MARINEMIX®, 

FORTY FATHOMS®, GP2 or 

equivalent) (available options)

Delete "FORTY 

FATHOMS®," from 

sentence.

...artificial sea salts (HW MARINEMIX®, GP2 or 

equivalent)....

FORTY FATHOMS®, a 

commercial product, is no 

longer available. Equivalent 

commercial products, such as 

HW MARINEMIX®, are 

acceptable replacements.

Section 14, Mysid, 

Mysidopsis bahia,

Survival, Growth, and 

Fecundity Test Method

1007.0, Section 14.5.16, 

Apparatus and 

Equipment, Page 217

Dissecting microscope (350-400X 

magnification)....

Replace "350-400X" with 

"35-40X."

Dissecting microscope (35-40X magnification).... Typographical error.

Section 14, Mysid, 

Mysidopsis bahia,

Survival, Growth, and 

Fecundity Test Method

1007.0, Section

14.10.2.3.1, Dilution 

Water, Page 228

Dilution water may be 

uncontaminated natural seawater 

(receiving water), HSB prepared 

from natural seawater, or artificial 

seawater prepared from FORTY 

FATHOMS® or GP2 sea salts (see 

Table 1 and Section 7, Dilution 

Water). 

Replace "FORTY 

FATHOMS®" with "HW 

MARINEMIX®" in 

sentence.

...or artificial seawater prepared from HW 

MARINEMIX® or GP2....

FORTY FATHOMS®, a 

commercial product, is no 

longer available. Equivalent 

commercial products, such as 

HW MARINEMIX®, are 

acceptable replacements. 

Consistency with Section 14, 

Table 3, Summary of Test 

Conditions..., Line 20, 

Dilution Water, Page 242.

Section 14, Mysid, 

Mysidopsis bahia,

Survival, Growth, and 

Fecundity Test Method

1007.0, Section 

14.10.1.3, Test 

Procedures, Page 227 

Eight replicate test vessels, each 

containing 5 to 7 day old animals, 

are used per effluent concentration 

and control.

Replace "5 to 7 day" with 

"5, 7-day."

Eight replicate test vessels, each containing 5, 7-day old 

animals, are used per effluent concentration and control.

Typographical error.

WET Methods Errata Sheet 23 October 2014
Errata for Methods for Measuring the Chronic Toxicity of Effluents and Receiving Waters to Marine and Estuarine Organisms, Third Edition. U.S. Environmental 

Protection Agency, Office of Water, Washington DC EPA/821/R–02/014, October 2002

Text Location Current Text Correction Being Made Corrected Text Reason For Correction 

Section 14, Mysid, 

Mysidopsis bahia,

Survival, Growth, and

Fecundity Test Method

1007.0, Section 

14.10.8.1.2, 

Observations During the 

Test, Page 229 

Temperature, pH, and salinity are 

measured at the end of each 24-hr 

exposure period in one test 

chamber at each test concentration 

and control. 

Add "beginning and"

between "measured at the"

and "end of each." Add 

"at least" between 

"exposure period in" and 

"one test chamber."

Temperature, pH, and salinity are measured at the 

beginning and end of each 24-hr exposure period in at 

least one test chamber at each test concentration and 

control. 

Clarification.

Section 14, Mysid, 

Mysidopsis bahia,

Survival, Growth, and 

Fecundity Test Method

1007.0, Section 

14.10.10.3.1, 

Termination of the Test, 

Page 230

Examine each replicate under a

stereomicroscope (240X) to 

determine the number of immature 

animals....

Replace "240X" with 

"24X."

Examine each replicate under a stereomicroscope (24X) 

to determine the number of immature animals....

Typographical error.

Section 14, Mysid , 

Mysidopsis bahia,

Survival, Growth, and 

Fecundity Test Method

1007.0, Section 14.11.1, 

Summary of Test 

Conditions and Test 

Acceptability Criteria, 

Table 3, Line 18, 

Dilution Water, Page 

242

Uncontaminated source of natural 

seawater, deionized water mixed 

with hypersaline brine or artificial 

sea salts (HW MARINEMIX®, 

FORTY FATHOMS®, GP2 or 

equivalent) (available options)

Delete "FORTY 

FATHOMS®," from 

sentence.

...artificial sea salts (HW MARINEMIX®, GP2 or 

equivalent)....

FORTY FATHOMS®, a 

commercial product, is no 

longer available. Equivalent 

commercial products, such as 

HW MARINEMIX®, are 

acceptable replacements.

Section 15 Sea Urchin, 

Fertilization Test 

Method, Section 

15.6.18.3.8 Consumable 

Materials, Table 1, 

Footnote 1, Page 301

Stir solutions 1 h to ensure that the 

salts dissolve. The salinity of the 

initial 840 mL of 100% effluent is 

adjusted to 30‰ by adding 25.2 g 

of dry artificial sea salts (FORTY 

FATHOMS®).

Replace "25.2 g of dry 

artificial sea salts (FORTY 

FATHOMS®)" with "an 

appropriate amount of dry 

artificial sea salts (e.g., 

HW MARINEMIX®) in 

sentence.

...by adding an appropriate amount of dry artificial sea 

salts (e.g., HW MARINEMIX®).

FORTY FATHOMS®, a 

commercial product, is no 

longer available. Equivalent 

commercial products, such as 

HW MARINEMIX®, are 

acceptable replacements.

WET Methods Errata Sheet 24 October 2014
Errata for Methods for Measuring the Chronic Toxicity of Effluents and Receiving Waters to Marine and Estuarine Organisms, Third Edition. U.S. Environmental 

Protection Agency, Office of Water, Washington DC EPA/821/R–02/014, October 2002

Text Location Current Text Correction Being Made Corrected Text Reason For Correction 

Section 15 Sea Urchin, 

Arbacia punctulata,

Fertilization Test 

Method 1008.0, Section

15.6.19.7, Test 

Organisms, Sea Urchins, 

Arbacia punctulata, 

Page 303

To successfully maintain about 25 

adult animals for 7 days at a field 

site, a screen-partitioned, 40-L 

glass aquarium using aerated, 

recirculating, clean saline water 

(30‰) and a gravel bed filtration 

system, is housed within a water 

bath, such as FORTY FATHOMS®

or equivalent (15°C). 

Replace "FORTY 

FATHOMS®" with "HW 

MARINEMIX® or 

equivalent" in sentence.

...is housed within a water bath, such as HW 

MARINEMIX® or equivalent (15°C). 

FORTY FATHOMS®, a 

commercial product, is no 

longer available. Equivalent 

commercial products, such as 

HW MARINEMIX®, are 

acceptable replacements.

Section 15 Sea Urchin, 

Arbacia punctulata, 

Fertilization Test 

Method 1008.0, Section

15.10.1.3.1, Dilution 

Water, Page 304

Dilution water may be 

uncontaminated natural seawater 

(receiving water), HSB prepared 

from natural seawater, or artificial 

seawater FORTY FATHOMS® or 

GP2 sea salts (see Table 2 and 

Section 7, Dilution Water).

Replace "FORTY 

FATHOMS®" with "HW 

MARINEMIX® or 

equivalent," in sentence.

...or artificial seawater HW MARINEMIX® or 

equivalent, or GP2 sea salts....

FORTY FATHOMS®, a 

commercial product, is no 

longer available. Equivalent 

commercial products, such as 

HW MARINEMIX®, are 

acceptable replacements.

Consistency with Section 15, 

Table 3, Summary of Test 

Conditions..., Line 11, 

Dilution Water, Page 308.

Section 15 Sea Urchin, 

Arbacia punctulata, 

Fertilization Test 

Method 1008.0, Section 

15.10.7.1.2, 

Observations During the 

Test, Page 306

Temperature, pH, and salinity are 

measured at the beginning of the 

exposure period in one test 

chamber at each concentration and 

in the control. 

Replace "of the exposure 

period ..." with "and end 

of each 24-hr exposure 

period in at least one test 

chamber at each test 

concentration and 

control."

Temperature, pH, and salinity are measured at the 

beginning and end of each 24-hr exposure period in at 

least one test chamber at each test concentration and 

control. 

Clarification. 

Section 15, Sea Urchin, 

Arbacia punctulata, 

Fertilization Test 

Method 1008.0, Section 

15.10.8.1, Start of the 

Test, Page 306

Sufficient test solution is prepared

at each effluent concentration to

provide additional volume for

chemical analyses, at the high,

medium, and low test

concentrations.

Replace "at the high, 

medium and low" with "all 

test."

Sufficient test solution is prepared at each effluent 

concentration to provide additional volume for chemical

analyses at all test concentrations.

Clarification. 

WET Methods Errata Sheet 25 October 2014
Errata for Methods for Measuring the Chronic Toxicity of Effluents and Receiving Waters to Marine and Estuarine Organisms, Third Edition. U.S. Environmental 

Protection Agency, Office of Water, Washington DC EPA/821/R–02/014, October 2002

Text Location Current Text Correction Being Made Corrected Text Reason For Correction 

Section 15 Sea Urchin, 

Arbacia punctulata, 

Fertilization Test 

Method 1008.0, Section

15.11.1, Summary of 

Test Conditions and Test 

Acceptability Criteria, 

Table 3, Line 11, 

Dilution Water, Page 

308 

Uncontaminated source of natural 

seawater; deionized water mixed 

with hypersaline brine or artificial 

sea salts (HW MARINEMIX®, 

FORTY FATHOMS®, GP2, or 

equivalent) (available options)

Delete "FORTY 

FATHOMS®," from 

sentence.

...artificial sea salts (HW MARINEMIX®, GP2 or 

equivalent)....

FORTY FATHOMS®, a 

commercial product, is no 

longer available. Equivalent 

commercial products, such as 

HW MARINEMIX®, are 

acceptable replacements.

Section 16, Red 

Macroalga, Champia 

parvula, Sexual 

Reproduction Test 

Method 1009.0, Section

16.10.1.3.1, Dilution 

Water, Page 343

The formula for the enrichment 

for natural seawater is listed in 

Table 1. Both EDTA and trace 

metals have been omitted. This 

formula should be used for the 2-

day exposure period, but it is not 

critical for the recovery period. 

Since natural seawater quality can 

vary among laboratories, a more 

complete nutrient medium (e.g., 

the addition of EDTA) may result 

in faster growth (and therefore 

faster cystocarp development) 

during the recovery period.

Add text from Line 12, 

Dilution Water, Table 3, 

Summary of Test 

Conditions..., Page 348

30‰ salinity natural seawater, or a combination of 50% 

of 30‰ salinity natural seawater and 50% of 30‰ 

salinity GP2 artificial seawater (see Section 7, Dilution 

Water). The formula for the enrichment for....

Consistency with Section 16, 

Table 3, Summary of Test 

Conditions, Line 12, Dilution 

Water, Page 348

Section 16, Red 

Macroalga, Champia 

parvula, Sexual 

Reproduction Test 

Method 1009.2, Section 

16.10.6.1.2, 

Observations During the 

Test, Page 344 

Temperature, pH, and salinity are

measured at the end of each 24-hr 

exposure period in at least one test 

chamber at each test concentration 

and in the control. 

Add "beginning and"

between "at the" and "end 

of." Add "at least"

between "period in" and 

"one test."

Temperature, pH, and salinity are measured at the 

beginning and end of each 24-hr exposure period in at 

least one test chamber at each test concentration and in 

the control. 

Clarification.

Appendix C, Dunnett's 

Procedure, Section 2.5, 

Computer Calculations, 

Page 408

The program was written in IBMPC FORTRAN by Computer 

Sciences Corporation, 26 W. 

Martin Luther King Drive, 

Cincinnati, OH 45268. A 

compiled version of the program 

can be obtained from EMSLCincinnati by sending a diskette 

with a written request.

Replace "...from EMSLCincinnati...." with 

replacement software 

information.

...can be obtained by clicking on "contact us" at 

http://www.epa.gov/med. Any software capable of 

calculating an analysis of variance (ANOVA) and a 

multiple of treatment means with the control mean 

(Dunnett's Procedure) can be used.

Computer program is no 

longer available from EMSLCincinnati. It can be obtained 

by sending a request to EPA 

using the link. Any software 

capable of completing the 

calculations is acceptable.

WET Methods Errata Sheet 26 October 2014
Errata for Methods for Measuring the Chronic Toxicity of Effluents and Receiving Waters to Marine and Estuarine Organisms, Third Edition. U.S. Environmental 

Protection Agency, Office of Water, Washington DC EPA/821/R–02/014, October 2002

Text Location Current Text Correction Being Made Corrected Text Reason For Correction 

Appendix H, Probit 

Analysis, Section 2, 

Page 436

A compiled, executable version of

the program and supporting 

documentation can be obtained 

from EMSL-Cincinnati by

sending a written request to

EMSL at 3411 Church Street,

Cincinnati, OH 45244.

Replace "...from EMSLCincinnati...." with 

replacement software 

information.

...can be obtained by clicking on "contact us" at 

http://www.epa.gov/med. Any software capable of 

calculating the EC1 and EC50 (or LC1 and LC50) and 

the associated 95% confidence intervals using Probit 

analysis can be used.

Computer program is no 

longer available from EMSLCincinnati. It can be obtained 

by sending a request to EPA 

using the link. Any software 

capable of completing the 

calculations is acceptable.

Appendix J, Trimmed 

Spearman-Karber 

Method, Section 10, 

Page 445

A computer program which

estimates the LC50 and associated

95% confidence interval using the 

Trimmed Spearman-Karber

Method, can be obtained through 

the EMSL, 3411 Church Street,

Cincinnati, OH 45244. The 

program can be obtained from 

EMSL-Cincinnati by sending a 

written request to the above 

address.

Replace "...through the 

EMSL-Cincinnati...."

with replacement software 

information.

...can be obtained by clicking on "contact us" at 

http://www.epa.gov/med. Any software capable of 

calculating the LC50 and associated 95% confidence 

interval using the Trimmed Spearman-Karber Method

can be used.

Computer program is no 

longer available from EMSLCincinnati. It can be obtained 

by sending a request to EPA 

using the link. Any software 

capable of completing the 

calculations is acceptable.

Appendix L, Linear 

Interpolation Method, 

Section 7.1, Page 458

A compiled, executable version of 

the program and supporting 

documentation can be obtained by 

sending a written request to

EMSL-Cincinnati, 3411 Church

Street, Cincinnati, OH 45244.

Replace "...sending a 

written request to EMSLCincinnati...." with 

replacement software 

information.

...can be obtained by clicking on "contact us" at 

http://www.epa.gov/med. Any software capable of 

calculating the mean and standard deviation of the 

bootstrapped ICp and associated 95% confidence 

intervals can be used.

Computer program is no 

longer available from EMSLCincinnati. It can be obtained 

by sending a request to EPA 

using the link. Any software 

capable of completing the 

calculations is acceptable.

Methods for Measuring the Acute Toxicity of Effluents and Receiving Waters to Freshwater and Marine Organisms, Fifth Edition. U.S. Environmental Protection Agency, Office 

of Water, Washington, DC EPA-821-R-02-012, October 2002. Short title: Acute Freshwater and Marine Manual.

Short-term Methods for Estimating the Chronic Toxicity of Effluents and Receiving Waters to Freshwater Organisms, Fourth Edition. U.S. Environmental Protection Agency, 

Office of Water, Washington, DC EPA/821/R–02/013, October 2002. Short title: Chronic Freshwater Manual.

Methods for Measuring the Chronic Toxicity of Effluents and Receiving Waters to Marine and Estuarine Organisms, Third Edition. U.S. Environmental Protection Agency, Office 

of Water, Washington DC EPA/821/R–02/014, October 2002. Short title: Chronic Marine and Estuarine Manual.

WET Methods Errata Sheet 27 October 2014