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

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- 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 specif ies :

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 2014Errata for Methods for Measuring th e 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

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- 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

an nually 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 o ut 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, C ereal leaves , and Trout chow (YCT)) which are used to culture and test organisms, the quality of food should mee t 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
concentrat ion 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 tox icant, 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 (X i ) 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 resu lts (X 1 ) 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 endpo ints (NOECs, IC25s, LC50s, etc.) should be plotted and examined to determine if the results (X 1 ) 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

.... Consi stency correction with Section 4. 1 6.2 in " Chronic Freshwater Manual " and

" Chronic Marine and Estuarine Manual ."

WET Methods Errata Sheet 2 October 2014Errata for Methods for Measuring th e 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

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- 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 LC 50 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 b e 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 2014Errata for Methods for Measuring th e 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

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- 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 s erving 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 valu es 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 ar ithmetic 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 " EMSL

Cincinnati hopes to

. ... " ...requiring aquatic toxicity tests. Standard reference materials

.... EM SL

- 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 2014Errata for Methods for Measuring th e 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

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- 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 equiva lent, 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 MARINE MIX ®

, are acceptable replacement s.

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 FATHOM S ® , 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 MARINE MIX ® , 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 equivale nt, 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 " FO RTY 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, i s no longer available. Equivalent commercial products, such as HW MARINE MIX ® , 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 MARINEMI X ® , or equivalent.... FORTY FATHOMS ® , a commercial product, is no longer available. Equivalent commercial products, such as HW MARINE MIX ® , are acceptable replacements.

WET Methods Errata Sheet 5 October 2014Errata for Methods for Measuring th e 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

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- 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 ® " wi th "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 MARINE MIX ® , are acceptable replacem ents.

Section 9, Acute Toxicity Test Procedures, Section 9.11.1, Feeding, Page 46 The organis m s are fed at test renewal, 48 h after the test is in itiate d, if Re gi on al or State
po licy re qu ires a 96 -h test du rati on

. Replace " fed at test renewal, 48 h after the test is i n itiate d" with " are provided food 2 h prior to test renewal at 48h ." The organis m s are provided food 2 h prior to test solution renewal at 48 h, if Re gi on al or State po licy

re qu ires a 96 h test du rati on

. Clarification of when foods can be add ed to match the text in the tables. The feeding regimes require that food(s) be mad e a v aila b le w h ile ho lding p ri o r to th e 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 th at the food is added 2 h p ri o r to test s olu ti o n renewal at 48 h ( reco mm ended); 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, M ysidopsis 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 MILLI

Q ® or equivalent deionized water ( see Section 7, Dilution Water); or receiving water (available options) Delete " FORTY FATHOMS ® ," from
sentence. ... or artificial sea salts ( HW MARINE MIX ®

, modified GP2, or equivalent).... FORTY FATHOMS ® , a commercial product, is no longer available. Equivalent commercial products, such as HW MARINE MIX ® , are acceptable replacements.

WET Methods Errata Sheet 6 October 2014Errata for Methods for Measuring th e 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

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- 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 S heepshead 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 hyper saline brine or artificial sea salts (HW MARINEMIX ® , FORTY FATHOMS ® , modified GP2, or equivalent) prepared with MILLI

Q ® or equivalent deionized water (see Section 7, Dilution Water); or receiving water (available options) Delete " FORTY FATHOMS ® ," from
sentence. ... or artificial sea salts ( HW MARINE MIX ®

, modified GP2, or equivalent).... FORTY FATHOMS ® , a commercial product, is no longer available. Equivalent commercial products, such as HW MARINE MIX ® , 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 1 8, 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 MILLI

Q ® or equivalent deionized water (see Section 7, Dilution Water); or

rece iving 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 MARINE MIX ® , are acceptable replacements.

WET Methods Errata Sheet 7 October 2014Errata for Methods for Measuring th e 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

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- 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, Sub

bullet 6, The Trimmed

Karber Method, Page 79 A co m puter pr ogram which

e sti m ates the LC 50 and associated 95% c onfidence in terval using the Trimmed

- Karber Method, can be ob tained th rough the E

nv i ron m ental Monitoring and Support Laboratory (E M SL), 26 W. M artin Lut h er King Dri v e, Cincinnati, OH 45268. T he
pr og ram can be ob tai n ed fr om EMSL

- Ci n ci nn ati by se nd ing a d is k ette with a written request to
t h e 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 . I t can be obtained by sending a request to EPA using the link. Any software capable of completing the calculations is
accep t ab l e.

Section 11, Acute Toxicity Data Analysis, Section 11.2.5.3, Sub

bullet 2, The Probit Method, Page 81 A m achine

- readable, co m p iled, version of a com puter program to esti m ate the LC1 and LC50 and associated 95% confi d ence in tervals using the Probit M ethod

can be obtained from EM SL

Ci n cinnati b y se nd ing a d is k ette with a written re qu est to th e E nv ir on men tal M on it o ri ng Syste m s La bo rat o ry, 2 6 W. M artin Lut h er King Dri v e, Cincinnati, OH 45268. Replace "

... from EMSL

Cincinnati...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 me t hod can be used. Computer program is no longer available from EMSL

Cincinnati. It ca n 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 " FORTY

FATHOMS ® " 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 MARINE MIX ® , are acceptable replacements.

W ET Methods Errata Sheet 8 October 2014Errata for Short

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– 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

enco untered 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 lea st 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 m eeting

m ini mum 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 Ma rine 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 2014Errata for Short

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– 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

av erage 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 aga inst 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 scale d 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

- an d- 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 concentrati on interval above and below the concentration
represe nting 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 describ ed in Section 4.14.5 and Section 9, Chronic Toxicity Tes t Endpoints and Data Analysis) the NOEC

LOEC interval is used for the

WET Methods Errata Sheet 10 October 2014Errata for Short

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– 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 a m m onia concentrations greater than 5 m g/L in the 100% eff l uent are an indicator that toxicity observed in the test m ay be due to a mm onia (USEPA, 1992). Replace " the 100% effluent are " with " in any concentration tested may be

. ... " Total a m m onia concentrations greater than 5 m g/L in any concentration tested may be an indicator that toxicity observed in the test m ay be due to a mm onia (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

v olume 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 measu red 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. Ins et " 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 sen tence. 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 2014Errata for Short

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– 02/013, October 2002 Text Location Current Text Correction Being Made Corrected Text Reason For Correction Section 12, Fathead Minnow

, Pimephales promelas, Embryo

Larval Survival and Teratogenicity Test Method 1001.0 , Section 12.3.5.1.2, Interferences, Pa ge 113 Total a m m onia concentrations greater than 5 m g/L in the 100% eff l uent are an indicator that toxicity observed in the test m ay be due to a mm onia (USEPA, 1992)

. Replace " the 100% effluent are " with " in any concentration tested may be "

... . Total a m m onia concentrations greater than 5 m g/L in any concentration tested may be an indicator that toxicity observed in the test m ay be due to a mm onia (USEPA, 1992). Clarification.

Section 12, Fathead Minnow

, Pimephales promelas, Embryo

Larval 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. Clarificatio n. 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, Embryo

Larval 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, Embryo

Larval 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 2014Errata 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 a m m onia concentrations greater than 5 m g/L in the 100% eff l uent are an indicator that toxicity observed in the test m ay be due to a mm onia (USEPA, 1992). Replace " the 100% effluent are " with " in any concentration tested may be ." Total a m m onia concentrations greater than 5 m g/L in any concentration tested may be an indicator that toxicity observed in the test m ay be due to a mm onia (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 s ample (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 2014Errata 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 m ale, 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 num ber of live young produced until eith er 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 shou ld not be included in the total numb er 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 o f 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. Clarifica tion. 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, ha rdness, 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, med ium, and low " with

" for all test ." Add /change word s " 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 2014Errata 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 th e program can be obtained from EMSL

- Cincinnati by sending a written request to EMSL at 3411 Church Street, Cincinnati, OH 45244. Replace "...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 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 EMSL

Cincinnati. 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 co m p iled, executab le version of the program can be obtained from EMSL

- Cincinnati by sending a written request to EMSL at 3411 Chu r ch Street, Cincinnati, OH 45244. Replace "...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 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 EMSL

Cincinnati. 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 co m puter program which esti m ates the LC50 and associated 95% confidence inter v al u si ng th e Tri mm ed Spear m an

- Karber Method, can be obtained from EMSL

- Cincinnati by sending a written request to EMSL, 3411 Church Street, Cincinnati, OH 45244. Replace "...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 int erval using the Trimmed Spearman

- Karber Method can be used. Computer program is no longer available from EMSL

Cincinnati. 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 T o obtain t h e program and supporti n g docu m entation, send a written request to EMSL

Ci n cinnati at 3411 Church Street, Cincinnati, OH 45244. Replace "...EMSL

Cincinnati. ..." with replacement softwar e 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 EMSL

Cincinnati. It can be obtained by sending a request to EPA using the link. Any software capable of completing the calculations is acceptable.

W ET Methods Errata Sheet 15 October 2014Erra ta 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

– 02/01 4 , 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 equiv alent" after "MARINEMIX ® " in
sentence. ....or artificial seawater prepared from commercial sea salts, such as HW MARINE MIX ® or equivalent if recommended in the method. FORTY FATHOMS ® , a commercial product, is no longer available. Equivalent commercial products, such as HW MARINE MIX ® , are acceptable replacements.

Section 4, Quality Assurance, Section 4.8.3, Food Quality, Page 11 ...the food should not be used (for analytical methods, s ee 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 organ isms, 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 Fo r food s (e.g.

, YCT ) whic h ar e

use d to cultur e.

... 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 2014Erra ta 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

– 02/01 4 , 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 lo garithm 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 char ts 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 o f 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 2014Erra ta 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

– 02/01 4 , 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 t hat are divergent results. For NOEC toxicity values, the control limits would consist of one concentration interval above and below the concentration
represe nting the central tendency. NOEC values cannot be averaged (Section 4.14.5); actual values and adja cent 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 Tes t Endpoints and Data Analysis) the NOEC

LOEC interval is used for the cusum chart. Reference: E nvironment 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 additiona l reference toxicants for future release. Standard reference materials...

. Delete sentence " EMSL

Cincinnati hopes to ..." . . .requiring aquatic toxicity tests. Standard reference materials

. ... EMSL

- Cincinnati no longer provides reference toxicants.

WET Methods Errata Sheet 18 October 2014Erra ta 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

– 02/01 4 , 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 FA THOMS ® , 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.

Sectio n 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 MARINE MIX ®

, 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 wate r. 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
sente nce. 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 MARINE MIX ® , are acceptable replacements.

WET Methods Errata Sheet 19 October 2014Erra ta 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

– 02/01 4 , 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 ® , o r equivalent, GP2, a natural seawater.... FORTY FATHOMS ® , a commercial product, is no longer available. Equivalent commercial products, such as HW MARINE MIX ® , are acceptable replacements.

Section 11, Sheepshead Minnow

, Cyprinodon variegatus, Larval Surviva l and Growth Test Method 1004.0

, Section 11.3.6.1.2, Interferences, Page 56 Total a m m onia concentrations greater than 5 m g/L in the 100% eff l uent are an indicator that toxicity observed in the test m ay be due to a mm onia (USEPA, 1992). Replace " the 100% effluent are " with " any concentration tested may be ." Total a m m onia concentrations greater than 5 m g/L in any concentration tested may be an indicator that toxicity observed in the test m ay be due to a mm onia (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 MARINE MIX ® , are acceptable replacements. Consistency with Section 11, Table 3, Summary of Te st 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 FATHOMS 7 ). Replace "136 g of dry artificial sea salts (FORTY FATHOMS 7 ) " 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 MARINE MIX ® , 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 concentrati on 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 cha mber at each test concentration and in the control. Clarification.

WET Methods Errata Sheet 20 October 2014Erra ta 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

– 02/01 4 , 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 FATHOMS 7 , GP2 or equivalent) (available options) Delete " FORTY FATHOMS 7

, " from
sentence. ...or artificial s ea salts ( HW MARINEMIX ®

, GP2 or equivalent).... FORTY FATHOMS ® , a commercial product, is no longer available. Equivalent commercial products, such as HW MARINE MIX ® , 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 salt s (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 MARINE MIX ® , 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 artific i al 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

comm ercial products, such as HW MARINE MIX ® , 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, O bservations 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 2014Erra ta 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

– 02/01 4 , 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 MARINE MIX ® , are acceptable replacements.

Section 13, Inland S ilverside, 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 S ilverside, 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 peri od 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 2014Erra ta 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

– 02/01 4 , October 2002 Text Location Current Text Correction Being Made Corrected Text Reason For Correction Section 13, Inland S ilverside

, 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 MARINE MIX ® , are acceptable replacements.

Section 14, Mysid

, Mysidopsis bahia

, Survival, Growth, and F ecundity 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 2014Erra ta 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

– 02/01 4 , 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 e ach ." 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

an imals

. ... 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 , Secti on 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 2014Erra ta 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

– 02/01 4 , October 2002 Text Location Current Text Correction Being Made Corrected Text Reason For Correction Section 15 Sea Urchin, Arbacia punc tulata

, 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, reci rculating, 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. E quivalent 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 Sufficien t tes t solutio n i s prepare d

a t eac h effluent concen t ra ti o n t o prov i d e add iti ona l vo l u m e fo r che m i ca l ana l y ses , a t t h e h i gh

, m

e d i u m , an d l o w te s t

concen t ra ti ons. Replace " at the high, medium and low " with " all
test ." Sufficien t tes t solutio n i s prepare d a t eac h effluent

concen t ra ti o n t o prov i d e add iti ona l vo l u m e fo r che m i ca l

ana l y ses at all te s t concen t ra ti ons. Clarification.

WET Methods Errata Sheet 25 October 2014Erra ta 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

– 02/01 4 , 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 MARI NEMIX ®

, 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 qua lity 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 T est 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 Sectio n 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 sali nity 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 IBM

PC FORTRAN by Computer Sciences Corporation, 26 W. Martin Luther King Drive, Cincinnati, OH 45268. A compiled version of the program can be obtained from EMSL

Cincinnati by sending a d iskette with a written request. Replace "...from 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 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 EMSL

Cincinnati. It can be obtained by sending a request to EPA using the link. Any software capable of completing the calculations is acc eptable.

WET Methods Errata Sheet 26 October 2014Erra ta 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

– 02/01 4 , October 2002 Text Location Current Text Correction Being Made Corrected Text Reason For Correction Appendix H, Probit Analysis, Section 2, Page 436 A co m p iled, executable version of
t h e program and supporting documentation can be obtained from EMSL

- Cincinnati by sending a written request to EMSL at 3411 Chu r ch Street, Cincinnati, OH 45244. Re place "... from 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 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 EMSL

Cincinnati. It can be obtained by sending a request to EPA using the link. Any software capable of completing the calculations is a cceptable. Appendix J, Trimmed Spearman

- Karber Method, Section 10, Page 445 A co m puter program which esti m ates t h e LC50 and associated 95% confidence inter v al u si n g t h e Tri mm ed Spear m an

- 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 a nd associated 95% confidence interval using the Trimmed Spearman

- Karber Method can be used. Computer program is no longer available from EMSL

Cincinna ti. 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
t h e program and supporti n g docu m entation can be obtained by sending a written request to EMSL

- Ci n cinnati, 3411 Church Street, Cincinnati, OH 45244. Replace "... sending a written request to 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 mean and standard deviation of the bootstrapped ICp and associated 95% confidence intervals can be used. Computer program is no longer available from EMSL

Cincinnati. 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 Estimat ing 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 Estua rine Manual.

WET Methods Errata Sheet 27 October 2014