EPA-600/R-98/182
January 1999
Errata for the Effluent and Receiving Water Toxicity
Testing Manuals: Acute Toxicity of Effluents and
Receiving Waters to Freshwater and Marine
Organisms; Short-Term Methods for Estimating the
Chronic Toxicity of Effluents and Receiving Waters to
Freshwater Organisms; and Short-Term Methods for
Estimating the Chronic Toxicity of Effluents and
Receiving Waters to Marine and Estuarine
Organisms.
U.S. Environmental Protection Agency
Office of Research and Development
Duluth, MN.
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ERRATA CITATION
USEPA, 1999. Errata for Effluent and Receiving Water Toxicity Test Manuals: Acute
Toxicity of Effluents and Receiving Waters to Freshwater and Marine Organisms;
Short-Term Methods for Estimating the Chronic Toxicity of Effluents and
Receiving Waters to Freshwater Organisms; and Short-Term Methods for
Estimating the Chronic Toxicity of Effluents and Receiving Waters to Marine and
Estuarine Organisms. January 1999. U.S. Environmental Protection Agency,
Office of Research and Development, Duluth, MN. EPA-600/R-98/182.
TABLE OF CONTENTS
Specific Errata for USEPA, 1993. Methods for Measuring the Acute Toxicity of Effluents and
Receiving Waters to Freshwater and Marine Organisms 3
Specific Errata for USEPA, 1994. Short-Term Methods for Estimating the Chronic Toxicity of
Effluents and Receiving Waters to Freshwater Organisms 7
Specific Errata for USEPA, 1994. Short-Term Methods for Estimating the Chronic Toxicity of
Effluents and Receiving Waters to Marine and Estuarine Organisms 13
EPA-600/R-98/182 January 1999
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Specific Errata for USEPA, 1993. Methods for Measuring the Acute Toxicity of Effluents and
Receiving Waters to Freshwater and Marine Organisms. Fourth Edition, August 1993. C.I. Weber
(Editor). U.S. Environmental Protection Agency, Environmental Monitoring Systems Laboratory,
Cincinnati, OH. EPA/600/4-90/027F.
SPECIFIC ERRATA:
1. REPLACE 4.4.1 ON PAGE 9 OF ACUTE MANUALS WITH THE FOLLOWING:
4.4 LABORATORY WATER USED FOR CULTURING AND TEST DILUTION
WATER
4.4.1 The quality of water used for test organism culturing and for dilution water used in
toxicity tests is extremely important. Water for these two uses should come from the
same source. The dilution water used in effluent toxicity tests will depend in part on the
objectives of the study and logistical constraints, as discussed in detail in Section 7,
Dilution Water. For tests performed to meet NPDES objectives, synthetic, moderately
hard water should be used. The dilution water used for internal quality assurance tests
with organisms, food, and reference toxicants should be the water routinely used with
success in the laboratory. Types of water are discussed in Section 5, Facilities, Equipment
and Supplies. 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. The
concentration of the metals Al, As, Cr, Cd, Cu, Fe, Pb, Ni, and Zn, expressed as total
metal, should not exceed 1 |ig/L each, and Cd, Hg, and Ag, expressed as total metal,
should not exceed 100 ng/L each. Total organochlorine pesticides plus PCBs should be
less than 50 ng/L (APHA, 1992). Pesticide concentrations should not exceed USEPA's
Ambient Water Quality chronic criteria values where available.
2. REPLACE PARAGRAPH 4.8.3 ON PAGE 10 OF THE ACUTE MANUAL WITH
THE FOLLOWING PARAGRAPH FROM THE CHRONIC MANUALS (4.8.3):
4.8.3 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. If the concentration
of total organochlorine pesticides exceeds 0.15 jig/g wet weight, or the concentration of
total organochlorine pesticides plus PCBs exceeds 0.30 jig/g wet weight, or toxic metals
(Al, As, Cr, Cd, Cu, Pb, Ni, Zn, expressed as total metal) exceed 20 jig/g wet weight, the
food should not be used (for analytical methods see AOAC, 1990 and USD A, 1989). For
foods (e.g., such as YCT) which are used to culture and test organisms, the quality of the
food should meet the requirements for the laboratory water used for culturing and test
dilution water as described in Section 4.4 above.
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H-.O.j JNcw Dcitcnes or rood used in cultunn^ cincl testing snould cilso DC cUicilyzecl tor toxic
or^cinics 3,110. metcils or wncncvcr diiiiculty is encountered in meeting niininiuni
cicceptciDility criteria, tor control survival cincl reproduction or ^rowtn. it tnc concenticition
OI totctl OlgctlllC ClllOlllie CXCCCQS 0.15 yu-g/g WCt WClgHt, Of tllC totctl COllCClltlcttlOll OI
or^cinocnlorinc pesticides plus Jr^i^lJs exceeds u. JU //^/^ wet wci^nt, or toxic mctcils
CXCCCQ 20 yU-g/g WCt WCl^llt, tllC lOOd SnOUld llOt DC USCQ (lOl ctllctlytlCctl lllCtllOuS SCC
AOAC, 1990; USD A, 1989).
3. CHANGE ON PAGE 43 IN REGARD TO HOLDING TIMES FOR EFFLUENT
SAMPLES:
8.5.4 Sample holding time begins when the last grab sample in a series is taken (i.e., when
a series of four grab samples are taken over 24-h period), or when a 24-h composite
sampling period is completed. If the data from the samples are to be acceptable for use in
the NPDES Program, the lapsed time (holding time) from sample collection to first use of
the sample in test initiation must not exceed 36 h. EPA believes that 36 h is adequate time
to deliver the samples to the laboratories performing the test in most cases. In the isolated
cases, where the permittee can document that this delivery time cannot be met, the
permitting authority can allow an option for on-site testing or a variance for an extension
of shipped sample holding time, directed to the USEPA Regional Administrator under 40
CFR 136.3(e) must include supportive data which show that the toxicity of the effluent
sample is not reduced (e.g., because of volatilization and/or sorption of toxics on the
sample container in no case should more than 72 h elapse between collection and first use
of the sample. In static-renewal tests, the original sample may also be used to prepare test
solutions for renewal at 24 h and 48 h and/or 72 h after test initiation, if stored at 4°C,
with minimum head space, as described in Subsection 8.5. Guidance for determining the
persistence of the sample is provided in Subsection 8.7.
4. CORRECT TYPOGRAPHICAL ERROR ON PAGE 49, SECTION 9.5.9.
Change paragraph 9.5.9, to read as follows:
9.5.8 Increases in pH may occur in test solutions during acute, static, and non-renewal
toxicity tests, resulting in an increase in the toxicity of pollutants such as ammonia. This
problem can be reduced by conducting the tests in a static-renewal or flow-through mode,
rather than a static non-renewal mode.
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5. CORRECT THE FOOTNOTES ON THE RECOMMENDED TEST SPECIES
TABLES
A. Section 5, Table 13: Cite appropriate reference for species formal name change from
Notropis leedsi to Cyprinella leedsi, Table 13, p. 61. Footnote reads:
1 Cyprinella leedsi (Bannerfish shiner, formerly Notropis leedsi; AFS, 1991) can be
used with the test conditions in this table, where it is the required test organism in
discharger permits.
B. Section 5, Table 15: Correct footnote for consistency with specific test conditions in
Appendix B, page 264 to indicate that specific alternate species can be used with the
test conditions in Table 15. The footnote reads:
1 Holmesimysis costata (mysid) can be used with the test conditions in this table,
except at a temperature of 12°C, instead of 20°C or 25°C, and a salinity of 32-34%o,
instead of 5-30%o, where it is the required test organism in discharge permits.
C. P. 264; Appendix B: For consistency in cross-referencing between Tables 13 and 15,
and Appendix B, The footnote number 1 on the table "Supplemental List of Acute
Toxicity Test Species" reads:
1 Test conditions for Cyprinella leedsi and Holmesimysis costata are found in Table
13, p. 61 and Table 15, p. 65, respectively.
6. LIST THE CORRECT SPECIES NAME IN SECTION 6.1.3 AND ADDS IT TO THE
REFERENCE SECTION.
A. Add the citation for the following reference on p. 27, paragraph 6.1.3 after the
beginning of the paragraph as follows:
6.1.3 The test species (AFS, 1991) listed in Subsection 6.1.2 are the recommended
acute toxicity test organisms. They are easily cultured in the laboratory, are sensitive
to a variety of pollutants, and are generally available throughout the year from
commercial sources. Summaries of test conditions for these species are provided in
Tables 11-17. Guidelines for culturing and/or holding the organisms are provided in
Appendix A.
B. Add reference to CITED REFERENCES, p. 119: AFS 1991 Common and
scientific names of fishes of the United States and Canada. Special Publ., 20 American
Fisheries Society, Bethesda, Maryland.
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7. REPLACE PARAGRAPH ITEM 6 OF 11.2.4.3 ON P. 84 WITH THE FOLLOWING
WHICH IS THE SAME TEXT IN THE CHRONIC MANUALS:
6. A computer program which estimates the LC50 and associated 95% confidence
interval using the Trimmed Spearman-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.
8. REPLACE WORDING IN ALL FOOTNOTES ON TABLE, P. 144. TABLE
ENTITLED "NUTRIENT STOCK SOLUTIONS FOR MAINTAINING ALGAL
STOCK CULTURES AND TEST CONTROL CULTURES.
Change the words from Stock #1 in footnotes a, footnote b, footnote c, footnote d, and
footnote e to Stock #2.
a ZnCl2 - Weigh out 164 mg and dilute to 100 mL. Add 1 mL of this solution to Stock #2.
b CoCl2.6H2O - Weigh out 71.4 mg and dilute to 100 mL. Add 1 mL of this solution to
Stock #2.
c Na2MoO4.2H2O - Weigh out 36.6 mg and dilute to 10 mL. Add 1 mL of this solution to
Stock #2.
d CuCl2.2H2O - Weigh out 60.0 mg and dilute to 1000 mL. Take 1 mL of this solution and
dilute to 10 mL. Take 1 mL of the second dilution and add to Stock #2.
e Na2SeO4 - Weigh out 119.6 mg and dilute to 100 mL. Add 1 mL of this solution to
Stock #2.
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Specific Errata for USEPA, 1994. Short-Term Methods for Estimating the Chronic
Toxicity of Effluents and Receiving Waters to Freshwater Organisms. Second Edition,
July 1994. Lewis, P., D. Klein in, J. Lazorchak, T. Norberg-King, M. Heber, and W.
Peltier (Eds). Environmental Monitoring Systems Laboratory, U.S. Environmental
Protection Agency, Cincinnati, OH. EPA/600/4-91/003.
SPECIFIC ERRATA:
1. REPLACE 8.3.4 ON PAGES 36 and 37 OF FRESHWATER CHRONIC MANUAL
WITH THE FOLLOWING SECTIONS.
8.3.4 THE FOLLOWING EFFLUENT SAMPLING METHODS ARE RECOMMENDED:
8.3.4.1 Continuous Discharges
8.3.4.1.1. If the facility discharge is continuous, a single 24-h composite sample is to be
taken.
8.3.4.2. Intermittent discharges
8.3.4.2.1 If the facility discharge is intermittent, a composite sample is to be collected for the
duration of the discharge but not more than 24 hours.
o.J.4.1 Continuous Discharges
T! It the facility discharge is continuous,, but the calculated retention time 01 a
continuously discnarged diluent is less tnan 14 clays and trie variability ol trie
waste is unknown, at a minimum, tour grab samples or tour composite
samples are collected over a -ZT—ri period. 1'or example, a grab sample is
taken every G h (total ol tour samples) and each sample is used tor a separate
toxicity test, or tour successive G-n composite samples are taKen and eacn is
used in a separate test.
2r It the calculated retention time ol a continuously discharged ellluent is
greater tnan IT- clays, or it it can be demonstrated tnat trie wastewater does
not vary more than 10/o in toxicity over a 24-h period, regardless ol
retention time, a single grab sample is collected tor a single toxicity test.
j. 1 lie retention time ot trie etiluent in trie wastewater treatment facility may be
estimated liom calculations based on the volume ol the retention basin and
rate ot wastewater miiow. However, trie calculated retention time may be
much greater than the actual time because ol short-circuiting in the holding
basin, VVnere snort-circuiting is suspected, or sedimentation may nave
reduced holding basin capacity, a more accurate estimate ol the retention
time can be obtained by carrying out a dye study.
o.J.'-r..Z. Intermittent discnarges
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o.J.'-k.Z.l it tne icicility disclicir^e is intermittent, 3, single ^iciD Sctmple is collected
niidwcty during eddi disclictige period. Jj/xdiiiples ot intermittent disclictiges die!
~nVVlien tlie ettluent is continuously discredited during d single o-li work sliitt
^one sdmple is collected) or two successive o-n WOIK sniits (jrvvo samples die
Collected).
z~! VVnen tlie tdcility retdins tne W3.stew^ter during 3.11 o-n WOIK sliitt, 3.nd tnen
tredts ctnd reledses tlie wdstewdter ds ct bdtcli disclidrj,e (one sdinple is
collected).
j. VVlien, ctt tlie end tlie sliitt, clectn up dctivities result in tlie disclidrj,e ot d slu^,
ot toxic Wcistes (^one S3.niple is collected).
2. SECTION 11: CHANGE "SEAWATER" TO "DEIONIZED" IN ITEM 4 OF
11.6.16.3:
11.6.16.3 Artemia nauplii are obtained as follows:
4. Drain the nauplii into a beaker or funnel fitted with a < 150 jim Nitex® or stainless
steel screen, and rinse with deionized water, or equivalent, before use.
3. SECTION 11.10.5 SHOULD BE CHANGED AS FOLLOWS:
11.10.5 FEEDING
11.10.5.1 The fish in each test chamber are fed 0.1 g (approximately 700 to 1000) of a
concentrated suspension of newly hatched (less than 24-h old) brine shrimp nauplii three
times daily at 4-h intervals or, as a minimum, 0.15 g are fed twice daily at an interval of 6
h. Equal amounts of nauplii must be added to each replicate chamber to reduce variability
in larval weight. Sufficient numbers of nauplii should be provided to assure that some
remain alive in the test chambers for several hours at the next feeding, but not in excessive
amounts which will result in depletion of DO below acceptable levels (below 4.0 mg/L)
4. ON PAGE 75, THE FIGURE LEGEND SHOULD BE:
Figure 2. Survival Mortality data for the fathead minnow, Pimephales promelas, larval
survival and growth test.
5. ON PAGE 81, CHANGE TABLE 2, REPLICATE D FOR AVG DRY WGT TO
0.254 RATHER THAN 0.508:
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TABLE 2. SUMMARY OF SURVIVAL AND GROWTH DATA FOR FATHEAD MINNOW,
PIMEPHALES PROMELAS, LARVAE EXPOSED TO A REFERENCE TOXICANT
FOR SEVEN DAYS1
Proportion of
NaPCP Survival in Replicate Mean Avg Dry Wgt (mg) In Mean
Cone. Chambers Prop. Replicate Chambers Dry Wgt
(|ig/L) A B C D Surv A B C D (mg)
0 1.01.00.90.9 0.95 0.7110.6620.6460.690 0.677
32 0.80.81.00.8 0.85 0.5170.5010.7230.560 0.575
64 0.9 1.0 1.0 1.0 0.975 0.6020.6690.6940.676 0.660
128 0.9 0.9 0.8 1.0 0.90 0.5660.6120.4100.672 0.565
256 0.70.91.00.5 0.775 0.4550.5020.6060.254 0.454
512 0.40.30.40.2 0.325 0.1430.1630.1950.099 0.150
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6. ON PAGE 97, TABLE 11 CHANGE THE ROW FOR MEAN (Yt), OLD
NUMBERS ARE SHOWN IN STRIKEOUT:
TABLE 11. FATHEAD MINNOW, PIMEPHALES PROMELAS, GROWTH DATA
NaPCP Concentration (\ieJD
Replicate Control 32 64 128 256 512
A
B
C
D
0
0.
0.
0
.711
.662
.646
.690
0
0.
0.
0
.517
501
723
.560
0
0.
0.
0
.602
669
694
.676
0
0.
0.
0
.566
612
410
.672
0
0.
0.
0
.455
502
606
.254
Mean(Y;)
S2
13 i
i 1
0.677 0.575 0.660 0.565 0.454 -
e'7'7 n FT r\ ccr\ r\ r^> 1 n con
.// U.DzD U.OOU U.024 U.38U
0.00084 0.010320.001620.012560.0218 -
23456
SECTION 11.3.3.7.6 ON PAGE 1U4: CHANGE THE SECTION TO EXPLAIN
THE DATA IN TABLE 17 CORRECTLY.
11.13.3.7.6 Since the purpose of this test is to detect a significant reduction in mean
weight, a one-sided test is appropriate. The critical value for this one-sided test is found in
Table 5, Appendix C. For an overall alpha level of 0.05, 15 degrees of freedom for error
and four concentrations (excluding the control) the critical value is 2.36. The mean weight
for concentration "/'" is considered significantly less than the mean weight for the control if
t; is greater than the critical value. Since t5 is greater than 2.36, the 256 |ig/L concentration
had significantly lower growth than the control. Hence the NOEC and the LOEC for
growth are 256 |ig/L and >256 |ig/L, respectively.
8. SECTION 13: CHANGES FOR CONSISTENCY OF TERMINOLOGY:
13.10.9 TERMINATION OF THE TEST
13.10.9.1 Tests should be terminated when 60% of the surviving control females have
produced their third brood, or at the end of 8 days, whichever occurs first. Because of the
rapid rate of development of Ceriodaphnia dubia, at test termination all observations on
organism survival and numbers of offspring should be completed within two hours. An
extension of more than a few hours in the test period would be a significant part of the
brood production cycle of the animals, and could result in additional broods.
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13.12 ACCEPTABILITY OF TEST RESULTS
13.12.1 For the test results to be acceptable, at least 80% of all control organisms must
survive, and 60% of surviving control females must produce three broods, with an average
of 15 or more young per surviving female.
Page 169: Table 3. Summary of Test Conditions and Test Acceptability Criteria for
Daphnid, Ceriodaphnia dubia, Survival and Reproduction Toxicity Tests with
Effluents and Receiving Waters (continued)
21. Test acceptability criteria:
80% or greater survival of all control organisms and an
average of 15 or more young per surviving female in the
control solutions. 60% of surviving control females must
produce three broods.
9. ON PAGE 170, SECTION 13.13.1.1, TABLE 4. CORRECT THE NUMBER OF
LIVE ADULTS IN 25% CONCENTRATION TO READ "2" AS SHOWN ON
PAGE 174 TO BE CONSISTENT WITH EXAMPLE CALCULATIONS.:
TABLE 4. SUMMARY OF SURVIVAL AND REPRODUCTION DATA FOR THE
DAPHNID, CERIODAPHNIA DUBIA, EXPOSED TO AN EFFLUENT FOR
SEVEN DAYS
Effluent
Concentration
Control
1.56
3.12
6.25
12.5
25.0
No.
of Young per Adult
Replicate
1
27
32
39
27
10
0
2
30
35
30
34
13
0
3
29
32
33
36
7
0
4
31
26
33
34
7
0
5
16
18
36
31
7
0
6
15
29
33
27
10
0
7
18
27
33
33
10
0
8
17
16
27
31
16
0
9
14
35
38
33
12
0
10
27
13
44
31
2
0
No.
Live
Adults
10
10
10
10
10
2 6-
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10. ON PAGE 175, CORRECT THE TYPOGRAPHICAL ERRORS IN THE
PERCENT TRIM AND THE LC50 ESTIMATE TO BE CORRECT WITH
DATA IN TABLE 7 ON PAGE 174.
TRIMMED SPEARMAN-KARBER METHOD. VERSION 1.5
DATE: 1 TEST NUMBER: 2
TOXICANT: effluent
SPECIES: Ceriodaphnia dubia
DURATION: 7 Days
RAW DATA: Concentration Number Mortalities
.00
1.25
3.12
6.25
12.5
25.0
SPEARMAN-KARBER TRIM:
Exposed
10
10
10
10
10
10
0
0
0
0
0
8
20.00 %
SPEARMAN-KARBER ESTIMATES: LC50: 19.28
95% CONFIDENCE LIMITS
ARE NOT RELIABLE.
NOTE:MORTALITYPROPORTIONSWERENOTMONOTONICALLYINCREASING.
ADJUSTMENTS WERE MADE PRIOR TO SPEARMAN-KARBER ESTIMATION.
Figure 5. Output for USEPA Trimmed Spearman-Karber program.
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Specific Errata for USEPA, 1994. Short-Term Methods for Estimating the Chronic
Toxicity of Effluents and Receiving Waters to Marine and Estuarine Organisms.
Third Edition. Klein in, D., G. Morrison, T. Norberg-King, M. Heber, and W. Peltier
(Eds). Environmental Monitoring Systems Laboratory, U.S. Environmental
Protection Agency, Cincinnati, OH. EPA 600/4-91-003.
SPECIFIC ERRATA:
1. REPLACE 8.3.4 ON PAGES 38 and 39 OF MARINE AND ESTUARINE
CHRONIC MANUAL WITH THE FOLLOWING SECTIONS.
8.3.4 THE FOLLOWING EFFLUENT SAMPLING METHODS ARE
RECOMMENDED:
8.3.4.1 Continuous Discharges
8.3.4.1.1. If the facility discharge is continuous, a single 24-h composite sample is to be
taken.
8.3.4.2. Intermittent discharges
8.3.4.2.1 If the facility discharge is intermittent, a composite sample is to be collected for
the duration of the discharge but not more than 24 hours.
o.J.4.1 Continuous Discharges
T! It the facility discharge is continuous,, but the calculated retention time 01 a
continuously cliscnargecl diluent is less tnan 14 clays and trie variability ot trie
waste is unknown, at a minimum, tour grab samples or tour composite samples
are collected over a .Z'-r-ri period. 1'or example, a grab sample is taKen every o n
(total ol lour samples) and each sample is used lor a separate toxicity test, or
tour successive o-n composite samples are taKen and eacn is used in a separate
test.
~£~. it trie calculated retention time ot a continuously cliscnargecl eiiiuent is greater
than 14 days, or 11 it can be demonstrated that the wastewater does not vary
more tnan lu/o in toxicity over a -ZT—ri period, regardless ot retention time, a
single grab sample is collected lor a single toxicity test.
j. 1 lie retention time ot trie eiiiuent in trie wastewater treatment facility may be
estimated irom calculations based on the volume ol the retention basin and rate
ot wastewater intiow. However, trie calculated retention time may be mucn
greater than the actual time because ol short-circuiting in the holding basin.
Vvnere snort-circuiting is suspected, or sedimentation may nave reduced noiclmg
basin capacity, a more accurate estimate ol the retention time can be obtained by
carrying out a dye study.
o.J.'4..z. intermittent cliscnarges
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o.J.4.2.1 It tnc ictcility disclictigC is intermittent, ct single gictb Sctiiiplc is collected midwcty
during C3.cn disclicir^c period. Jj/xciiTiplcs or intermittent uisclicir^cs circ!
VVncn tnc diluent is continuously disclicir^ed during 3, single o-n WOIK sniit
Sctniple is collected) or two successive o-n work sniits (two Sctniples die
2"; VVnen tne ictcility ictctiiis tnc wdstcwdtcr during dii o-n WOIK snitt, diid tncii
3.110. rdC3.SCS tllC W3.StCW3.td clS 3, DdtCll dlSCndl^C (^OllC ScUTlplC IS COllCCtCdJ.
j. VVncii, ctt tnc cud tnc snitt, clCctn up activities result in tnc uiscndr^c or d sluj, or
TOXIC W3.STCS (^OllC SdlllplC IS COllCCtCdJ.
PAGE 105, SECTION 11.13.3.5.6: SHAPIRO-WILK'S CRITICAL VALUE
SHOULD BE 0.844 NOT 0.876:
1 1. 13.3.5.6 The decision rule for this test is to compare W with the critical value found in
Table 6, Appendix B. If the computed W is less than the critical value, conclude that the
data are not normally distributed. For this example, the critical value at a significance
level of 0.01 and 16 observations (n) is 0.844 0^76. Since W = 0.938 is greater than the
critical value, the conclusion of the test is that the data are normally distributed.
PAGE 158, SECTION 12.13.2.9.1: TABLE 10 MORTALITY DATA IS
CORRECTED TO BE CONSISTENT WITH DATA IN TABLE 3 ON PAGE 149
AND A NEW FIGURE 5 FOR PAGE 159 IS PROVIDED THAT GIVES THE
CORRECT VALUES FOR PROBIT CALCULATION USING THE SAME
VERSION AND PROGRAM USED BEFORE.
TABLE 10. DATA FOR PROBIT ANALYSIS
_ SDS Concentration (mg/L)
Control
0.5
1.0
2.0
4.0
8.0
Number Dead 24 54
Number Exposed 40 40
42
40
108
40
32
40
40
40
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USEPA PROBIT ANALYSIS PROGRAM
USED FOR CALCULATING LC/EC VALUES
Version 1.5
Probit Analysis of Sheepshead Minnow Embryo-Larval Survival and
Teratogenicity Data
Cone.
Number
Exposed
Control
0.5000
1 .0000
2.0000
4.0000
8.0000
40
40
40
40
40
40
Number
Resp.
Observed
Proportion
Responding
2
5
4
10
32
40
0
0
0
0
.5000
.1250
.1000
.2500
0.8000
1
.0000
Proportion
Responding
Adjusted for
Controls
0.0000
0.0369
0.0094
0.1745
0.7799
1.0000
Chi - Square for Heterogeneity (calculated) = 0.782
Chi - Square for Heterogeneity (tabular value) = 7.815
Probit Analysis of Sheepshead Minnow Embryo-Larval Survival
and Teratogenicity Data
Estimated LC/EC Values and Confidence Limits
Point
Exposure Lower Upper
Cone. 95% Confidence Limits
LC/EC 1.00
LC/EC 50.00
1.187
2.912
0.643
2.432
1.601
3.361
Figure 5. Output for USEPA Probit Program, Version 1.5.
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January 1999
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4. PAGE 205, SECTION 13.13.3.4: In Table 12, Replicate C in 25% is in error,
should read 0.079 rather than 1.079 as given.
5. PAGE 212, SECTION 13.13.3.7.5: In Table 18, the t value for the 6.25%
concentration should read -0.120 rather than 0.170.
6. PAGE 251, TABLE 4: Data for 100 ppb and Total mysids for replicate 8 should
read 5 not 4.
7. PAGE 416, APPENDIX B.3.3: Table B.7 Replicate 1 of the 25% should be 0.873.
The mean is 0.882, the squared of the standard deviation is 0.0024.
8. PAGE 418, CHANGES ARE GIVEN IN REDONE TO CORRECT THE
TYPOGRAPHICAL ERRORS.
4.2.4.1 For RPs greater than zero or less than one:
Angle (radians) = arc sine \/RP
Example: If RP = 0.60:
Angle = arc sine v/0.60
= arc sine 0.7746
= 0.8861 radians
4.2.4.3 Modification of the arc sine square root when RP = 10
Angle = 1.5708 radians - (radians for RP = 0)
Example: Using above value:
Angle = 1.5708-0.1120= 1.4588 radians
9. PAGE 435, APPENDIX D.3.6: Table D.4 T statistic for the 50% is -0.592 and
should be +0.592.
EPA-600/R-98/182 January 1999
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